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15 June 2016, Volume 44 Issue 6 Previous Issue    Next Issue
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Cell and Molecular Biology

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Cell and Molecular Biology

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Establishing and identification of an immortal cell line of familial papillary thyroid carcinoma with MLH1 R217C/BRAF V600E mutations HAO Weijing1, YU Yang1, WANG Qingsong2, ZHAO Li3, YE Yan4, DONG Li1, LI Jiansen5, MENG Xiangrui5,GAO Ming1 2016, 44 (6):  657-661.  doi: 10.11958/20150376 Abstract ( 1519 )   PDF (668KB) ( 4105 )   Abstract: Objective To establishing an immortal cell line of familial papillary thyroid carcinoma (FPTC), and explore a new approach for studying familial non-medullary thyroid carcinoma (FNMTC). Methods The specimen from a patient with FPTC was selected, separated, and the primary cells were cultured using DMEM/F12 medium (with TSH, T3,EGF and hydrocortisone). To inducing cell immortalization, the exogenous genes SV40T/TERT were transfected into cells by two ways. RT-PCR was used to detect the expressions of thyroid peroxidase (TPO), thyroid globulin (TG), thyroid stimulating hormone receptor (TSHR) and sodium/iodide co- transporter (NIS). Immunofluorescence method was used to detect the expressions of TPO and GPC3. In order to detect the genomic mutations, the peripheral blood DNA of the patient wasextracted. The cell genome was detected. Results The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells can stably grow for six months, FPTC-S (with SV40T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST (with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST can stably express TPO,TG and TSHR in mRNA level. MLH1 R217C mutation existed in the peripheral blood of the patient, and BRAF V600E mutation existed in the primary cultured cells. Either the primary or the immortal cells showed MLH1 R217C mutation.Conclusion This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH1 R217C and BRAF V600E mutations. This cell line provides a research model for studying these mutations in FPTC. Related Articles | Metrics

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Identification and application of sphingosine-1-phosphate receptor of human umbilical vein endothelial cells LIANG Yun1, LUO Guanghua2, LIU Hong1, YU Miaomei2, WANG Min1, LIU Hongyao1,HUANG Bo1, XU Ning3, ZHANG Xiaoying1△ 2016, 44 (6):  662-664.  doi: 10.11958/20160032 Abstract ( 859 )   PDF (953KB) ( 3689 )   Abstract: Objective To identify the expression of sphingosine-1-phosphate receptor (S1PR) subtypes, C-myc and His tag proteins of human umbilical vein endothelial fusion cell line, EA.hy926 and human umbilical vein endothelial cells(HUVEC), CRL-1730 for studying the function of apolipoprotein M (ApoM)-S1P axis. Methods Two kinds of cells (EA.hy926 and CRL-1730) were cultured to reach the density of 60%-70% in vitro. Immunofluorescence technique was employed to investigate the expressions of coagulation factor Ⅷ (FⅧ), ApoM, S1PR1-S1PR5, C-myc and His tag proteins. Results (1) Two kinds of cells both expressed FⅧ and ApoM. FⅧ presented scattered particle distribution in CRL-1730,while uniform distribution in EA.hy926. However, ApoM was strongly expressed and widely distributed in cytoplasm of twokinds of cells. (2) S1PR1-3 can be detected on their membrane other than S1PR4 and S1PR5. S1PR1 was highly expressed but S1PR2 and S1PR3 were in a low level expression. (3) Two kinds of cells both expressed C-myc and His tag proteins in cytoplasm. Conclusion Two kinds of cells have the properties of endothelial cells and can express FⅧ, ApoM, C-myc and His tag proteins. It is not suitable for choosing C-myc and/or His tag–conjugated recombinant ApoM to study the fuction of ApoM-S1P axis with these two kinds of cells. Related Articles | Metrics

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Study on changes of elastic modulus of injured astrocytes detected by atomic force microscope CHEN Miaobin1, LI Xiaohong2, WU Sen3, WANG Jingjing2, SUN Hongtao2 2016, 44 (6):  665-668.  doi: 10.11958/20160059 Abstract ( 1130 )   PDF (1618KB) ( 3806 )   Abstract: Objective To investigate changes of the elastic modulus of astrocytes induced by injury. Methods The astrocytes were isolated and extracted from the 2-day old SD rats, and identified by immunofluorescence staining with glial fiber acidic protein (GFAP) antibody. Cells were divided into control group and injured group. The injured group was astrocytes 6 h after being injured by the cell damage instrument. The control group was astrocytes without any injury. The elastic modulus in liquid phase was tested by atomic force microscope in two groups. Results were compared and analyzed between two groups. Results The purification rate of rat astrocytes was more than 95% . Six hours after the injury, the astrocytes were in disorder, and some of cell bodies were swelling. The mechanical topographic maps and force indentation curves were obtained. The elastic modulus of astrocytes was significantly increased in injured group compared with that of control group［(1 689±693) Pa vs. (724±283) Pa, P＜0.01]. Conclusion The injury stimulus increases the elastic modulus of astrocytes, which provides theoretical basis for understanding intracranial physical microenvironment after traumatic brain injury in animal experiments. Related Articles | Metrics

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The effect of LPS on VEGF expression in microglia ZENG Yong, LYU Jingfang, YU Gongjie, GAO Weiwei, ZHANG Jianning 2016, 44 (6):  669-671.  doi: 10.11958/20150374 Abstract ( 962 )   PDF (1475KB) ( 4209 )   Abstract: Objective To investigate whether lipopolysaccharide (LPS) can induce vascular endothelial cell growth factor(VEGF) expression in microglia regulated by hypoxia inducible factor-1α (HIF-1α). Methods The cultured BV2 cells were divided into four groups: control group, LPS (100 µg/L) simulated group, LPS (100 µg/L) + LPS antagonist (LRS, 200 µg/L)intervened group and LPS (100 µg/L) + HIF-1α inhibitors FM19G11 (10 mmol/L) intervened group. Immunofluorescence staining,Western blotting and ELISA were used to detect the expressions of VEGF and HIF-1α. Results Compared with the control group, the VEGF expression level was obvious high in LPS simulated group (P < 0.05). LRS inhibited this effect of LPS (P <0.05). The HIF-1α level was increased in LPS simulated group at 8 h post-injury (P < 0.05). FM19G11, the inhibitor of HIF- 1α reduced the expression of VEGF induced by LPS (P < 0.05). Conclusion LPS can up-regulate the expression of VEGF by HIF-1α. Related Articles | Metrics

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Effects of carbon monoxide on lipopolysaccharide induced damage in rat alveolar macrophages LIU Wei, YU Jianbo, WANG Dan, SHI Jia 2016, 44 (6):  672-674.  doi: 10.11958/20150377 Abstract ( 906 )   PDF (346KB) ( 5066 )   Abstract: Objective To evaluate effects of carbon monoxide (CO) on lipopolysaccharide (LPS) induced damage and possible mechanism in rat alveolar macrophages. Methods Rat alveolar macrophages were cultured in DMEM containing 10% fetal bovine serum with 5% CO2 at 37 ℃ in Heraeus sepatech. The cells were divided into four groups using random number table (n=10): control group (group C)， CO group, LPS group and LPS + CO group. The CO release molecule- 2(CORM- 2) 100 μmol/L was added into CO group， LPS 10 mg/L was added into LPS group, cells were pretreated with CORM-2 100 μmol/L for 1 h then LPS 10 mg/L was added into LPS+CO group, the same amount of PBS was added to group C. Proliferation was measured by MTT assay. Apoptosis and mitochondrial membrane potential were detected with flow cytometer. The content of ATP was tested by ATP content kit. Drp1 mRNA was measured by RT-PCR, and Drp1 expression was determined by Western blot assay. Results Compared with group C, the cell vitality, content of ATP and mitochondrial membrane potential were decreased in LPS group and LPS + CO group， and cell apoptosis rate, Drp1 mRNA and protein expression were increased (P < 0.05). There were no significant changes were found in CO group. Compared with LPS group, the cell vitality, content of ATP and mitochondrial membrane potential were increased in LPS + CO group， and the cell apoptosis rate, Drp1 mRNA and protein expression were decreased (P < 0.05). Conclusion Carbon monoxide can alleviate LPS- induced damage in rat alveolar macrophages, which is related with downregulation of Drp1 and amelioration of mitochondrial function. Related Articles | Metrics

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The study on the role of salinomycin in the proliferation and epithelial-mesenchymal transition of breast cancer MCF-7 mammosphere cells MA Heyao， FU Yingzi， HE Miao， YAN Yuanyuan， JIANG Qian， SUN Yezhi, WEI Minjie 2016, 44 (6):  675-678.  doi: 10.11958/20150221 Abstract ( 1029 )   PDF (514KB) ( 4085 )   Abstract： Objective To investigate the effects of salinomycin on the cell proliferation and epithelial-mesenchymal transition (EMT) of MCF-7 mammosphere (MCF-7 MS). Methods Breast cancer MCF-7 cells were cultured in suspension in serum-free medium to obtain MCF-7 MS. The cell viability of MCF-7 MS cells treated with serial concentrations of 0, 10, 30,100, 300, 1 000, 3 000 and 10 000 nmol/L of salinomycin for 24 hours were detected by CCK-8 assay. The half maximal inhibitory concentration (IC50) was calculated. Western blot analysis was performed to detect the expression levels of E-cadherin and Snail in MCF-7 MS cells treated with 30 nmol/L and 60 nmol/L salinomycin. The same capacity of DMSO was added to MCF-7 MS as control group. The xenograft tumors from MCF-7 MS transplant mice were divided into control group (the same capacity of normal saline) and salinomycin group (5 mg/kg salinomycin), then the expressions of E-cadherin and Snail were dectected by immunohistochemical staining. Results With the increased concentration of salinomycin, the cell survival rate of MCF-7 MS cells decreased (P < 0.05). The IC50 after 24 h-treatment was 989 nmol/L. Both 30 and 60 nmol/L of salinomycin increased the expression of E-cadherin and decreased the expression of Snail compared with control group. In addition, 60 nmol/L treatment group showed more significant effect (P < 0.05). In xenograft tumors from MCF-7 MS transplant mice, the expression of Snail decreased, and E-cadherin increased in salinomycin treatment group compared with control group (P < 0.01). Conclusion Salinomycin can inhibit the cell proliferation and EMT in MCF-7 MS cells, which is a potential drug to target cancer stem cells. Related Articles | Metrics

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Effects of RNAi targeting CDX2 gene expression on biological characters of human leukemia cells and its relevant mechanisms SUN Weimei1, LI Jianchang1, JIA Xiuhong1, LI Youjie2, TANG Shenhua1 2016, 44 (6):  679-682.  doi: 10.11958/20150299 Abstract ( 845 )   PDF (3504KB) ( 3856 )   Abstract: Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase- 9, Bax mRNA and protein were tested by RTPCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase- 9 and Bax mRNA and protein were significantly higher than those of other two groups (P＜0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the downregulation of BCR-ABL and the up-regulation of caspase-9 and Bax. Related Articles | Metrics

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Interferon-γ promotes immunomodulatory of adipose tissue-derived mesenchymal stem cells on peripheral blood lymphocytes WANG Ping1, GU Xin2, ZHANG Na3, ZHANG Hong1, SHI Shuainan4, WANG Yuliang4 2016, 44 (6):  683-686.  doi: 10.11958/20150261 Abstract ( 1251 )   PDF (389KB) ( 3801 )   Abstract： Objective To investigate the immunomodulatory effects and the mechanism of interferon (IFN)-γ-pretreated adult autologous adipose tissue-derived mesenchymal stem cells (ADSCs) on peripheral blood lymphocytes. Methods ADSCswere obtained from adult subcutaneous adipose tissues. IFN- γ with and without pretreated ADSCs were used as IFN- γ-pretreated group and IFN-γ-unpretreated group, which were cultured with autologous peripheral blood mononuclear cells (PBMCs) at different concentrations of ADSCs- to- PBMCs ratios in presence of concomitant phytohemagglutinin (PHA)/interleukin (IL)-2 stimulation. After 5 days of culture, the proliferatory inhibitory rate of activated T cells, the percentage of CD4 + CD25 + regulatory T cells (Treg), and the expression of indoleamine 2,3- dioxygenase (IDO) mRNA were assessed.Results ADSCs were isolated from autologous adipose tissue, which strongly expressed CD73, CD90, and CD105, as well as displayed adipogenic and osteogenic differentiation. The percentage of CD4+CD25+Treg was significantly higher in IFN-γ-pretreated group than that of IFN-γ-unpretreated group. The expression level of IDO mRNA in ADSCs was significantlyincreased in IFN-γ-pretreated group than that of IFN-γ-unpretreated group. The proliferation inhibition of activated T cells was significantly decreased in IDO-blocker group than that of IFN- γ-pretreated group (P < 0.01). Conclusion These results suggest that IFN- γ can promote immunosuppressive effects of ADSCs on activated T cells through increased expression of IDO. Related Articles | Metrics

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The inhititory effects of allicin on human lung epithelial cell apoptosis induced by porphyromonas gingivalis MENG Tiantian, LI Xin 2016, 44 (6):  687-690.  doi: 10.11958/20150281 Abstract ( 1346 )   PDF (463KB) ( 3687 )   Abstract： Objective To investigate the influence of different concentrations of allicin in apoptosis of A549 cells induced by porphyromonas gingivalis (P.gingivalis). Methods The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of allicin in inhibiting P.gingivalis were investigated by broth dilution methods. The tetrazolium salts (MTT) assay was used to detect the viability of A549 cells infected by P.gingivalis and treated with different concentrations of allicin (64 mg/L, 96 mg/L and 128 mg/L). The flow cytometry FITC/PI staining was used to detect apoptotic rates of A549 cells treated by P.gingivalis and/or allicin for 24 h. Results The values of MIC and MBC of allicin for inhibiting P.gingivalis were 64 mg/L and 128 mg/L respectively. MTT assay showed that the cell viability was significantly increased with the increased concentration of allicin in a concentration- dependent manner (P < 0.05). There was no significant difference in cell viability between allicin (128 mg/L) group and control group, which showed that allicin was no significant cytotoxicity in A549 cells. Flow cytometry assay showed that the apoptotic rates from high to low were P.gingivalis group > P. gingivalis + allicin group > allicin group > control group with significant differences (P < 0.01). The apoptosis of A549 cells induced by P.gingivalis was significantly inhibited by allicin (P < 0.01). Conclusion Allicin can inhibit P.gingivalis infection in lung epithelial cells. There is a good prospect in the application of allicin in the treatment of pulmonary infection in patients with periodontitis. Related Articles | Metrics

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A correlation study of blood-brain barrier permeability changes and EBA, VEGF expression in rats following radiation-induced brain injury CHEN Bolin1, SUN Yi2, LIANG Bin3, JIN Xuelong1 2016, 44 (6):  691-693.  doi: 10.11958/20150439 Abstract ( 965 )   PDF (917KB) ( 3866 )   Abstract: Objective To explore the dynamic changes of endothelial barrier antigen (EBA) and vascular endothelial growth factor (VEGF) expressions in cerebral cortex under the condition of blood-brain barrier damage in rats following radiation-induced brain injury, which provided clinical references. Methods Forty-eight clean grade male SD rats were divided into the control group and 7 d, 14 d, 28 d after brain irradiation group (n=12 for each group) by using stochastic indicator method. The radiation-induced brain injury model was established by using electronic computer X-ray tomography technique. The 3% Evans blue (EB) was injected into rats according to the dose of 3 mL/kg via the tail vein, then the blood vessels of cerebral cortex were exposed after having a craniotomy. EB extravasation was detected by microcirculation microscope. The permeability of blood-brain barrier was evaluated by using microscope vascular camera device. The expressions of EBA and VEGF in the cerebral cortex were measured by immunohistochemistry staining in each group. Results Both of EB extravasation and VEGF expression in rat cerebral cortex were significantly increased in injury group at day 7, 14 and 28 after brain irradiation compared with those of control group (P＜0.05), and which were gradually decreased from day 7 to day 28 after brain irradiation. There were significant differences in EB extravasation and VEGF expression between the injury subgroups (P＜0.05). There was a positive correlation between EB extravasation and VEGF expression (r=0.898, P < 0.001).The expression levels of EBA were decreased at different time points in injury groups compared with those of control group(P＜0.05), and gradually increased from day 7 to 28 after injury. There were significant differences in expression levels ofEBA between injury subgroups (P＜0.05). The expression of EBA was negatively correlated with EB extravasation (r=-0.866,P < 0.001). Conclusion The increases of blood-brain barrier permeability have important relation to the decreases of EBA expression and the increases of VEGF expression after radiation-induced brain injury. Related Articles | Metrics

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The potential effects of EPCs-angiogenesis on ischemia-reperfusion injury ZHANG Zhishen, DONG Beibei, XIE Keliang, YU Yonghao 2016, 44 (6):  694-699.  doi: 10.11958/20160042 Abstract ( 1012 )   PDF (3280KB) ( 4244 )   Abstract：Objective To explore the potential effects of endothelial progenitor cells (EPCs)- angiogenesis on mechanism of alleviating cognitive dysfunction in rats subjected to cerebral ischemia-reperfusion (I/R) injury. Methods Atotal of 121 male Sprague–Dawley (SD) rats were randomly divided into four groups: Sham group (n=31), focal I/R(MCAO,0.9% saline 10 μL, n=30) group, MCAO+Vehicle (sodium azide, 0.1% Vehicle 10 μL, n=30) group and MCAO+HPX (1.86 g/L HPX 10 μL, n=30) group. The modified neurological severity scores (mNSS) was carried out to determine neurological function deficit after I/R. Morris water maze (MWM) was carried out to assess learning and memory abilities after I/R. The circulating EPCs after I/R were counted by flowcytometry (FCM) combined with double- immunofluorescence staining of CD34 and CD133. Angiogenesis in rat penumbra cortex after I/R was assessed by immunohistochemical technique combined with immunofluorescent chromogenic detection of CD31 and vWF. Results Compared with sham group, the mNSS scores, the escape latency and the circulating EPCs count were increased after I/R, the time percentage spent in target quadrant was reduced, and the new vessel density in penumbra cortex was increased after I/R in MCAO group (P < 0.05 respectively).There were no significant differences in mNSS score, the escape latency, the time percentage spent in target quadrant, the circulating EPCs count and the new vessel density in penumbra cortex between MCAO group and MCAO+Vehicle group (P >0.05). The mNSS score and the escape latency were significantly decreased, the circulating EPCs count and new vessel density in penumbra cortex were significantly increased after I/R in MCAO+HPX group compared with those of MCAO+Vehicle and MCAO group (P < 0.05). Conclusion EPCs-angiogenesis signaling plays positive effects on HPX alleviating cognitive dysfunction in rats subjected to focal cerebral ischemia reperfusion injury. Related Articles | Metrics

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Influence of dexmedetomidine on expressions of PKCγ, CaMKⅡα and pCaMKⅡα in spinal cord in rats with hyperalgesia SUN Zhe, WANG Zhifen, HE Ying, WANG Guolin, YU Yonghao, YUAN Yuan 2016, 44 (6):  700-703.  doi: 10.11958/20150404 Abstract ( 1101 )   PDF (375KB) ( 3915 )   Abstract： Objective To investigate the influence of dexmedetomidine on expressions of protein kinase c (PKC)γ, calcium/calmodulin-dependent protein kinase (CaMK)Ⅱα and pCaMKⅡα in spinal cord in rats with incisional pain (IP) and remifentanil-induced hyperalgesia. Methods Forty male Sprague-Dawley rats, aged 2-3 months, weighing 240-260 g,were randomly divided into 5 groups (n=8 each): blank control group (group C), remifentanil+incisional pain group (group R+I), dexmedetomidine + remifentanil + incisional pain group (group D+R+I), dexmedetomidine + remifentanil + incisional pain + phorbol myristate acetate + DMSO group (group D+R+I+P+DMSO) and dexmedetomidine + remifentanil + incisional pain + DMSO group (group D+R+I+ DMSO). The incisional pain rat model was established by a plantar incision in left hind paw. Remifentanil was infused at a rate of 1.2 μg·kg-1·min-1 for 90 min via the caudal vein. Dexmedetomidine was adminis⁃ tered subcutaneously at a dose of 50 μg/kg at 30 min before plantar incision. Phorbol myristate acetate and DMSO were intrathecally injected at a dose of 10 μL. Paw withdrawal latency (PWL) to thermal stimulation and paw withdrawal threshold (PWT) to von Frey hair stimulation were measured 24 h before remifentanil infusion (T0) and at 2, 6, 24 and 48 h (T1-4) after intraveonus remifentanil injection. The rats were sacrificed after the last behavioral test and the L4- 6 segment of spinal cord was removed to determine the expressions of PKCγ, CaMKⅡα and pCaMKⅡα in spinal cord by Western blot analysis. Re⁃sults Compared with group C, the value of PWL was significantly shortened and PWT was significantly decreased except T0, and the expressions of PKCγ, CaMKⅡα and pCaMKⅡα were up-regulated in other groups. Compared with group R+I,the value of PWL was significantly prolonged and PWT was significantly increased, the expressions of PKCγ, CaMKⅡα and pCaMKⅡα were down-regulated in group D+R+I and group D+R+I+DMSO. Compared with group D+R+I, the value of PWL was significantly shortened and PWT was significantly decreased, the expressions of PKCγ, CaMKⅡα and pCaMKⅡα were up-regulated in group D+R+I+P+DMSO. Compared with group D+R+I+P+DMSO, the value of PWL was significantly prolonged and PWT was significantly increased, the expressions of PKCγ, CaMKⅡα and pCaMKⅡα were down-regulated in group D+R+I+DMSO. Conclusion Dexmedetomidine can reduce the expressions of PKCγ, CaMKⅡα and pCaMKⅡα in spinal cord in rats with IP and hyperalgesia induced by remifentanil. Related Articles | Metrics

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Effects of receptor interacting protein (RIP)1 on isoniazid induced hepatocyte necroptosis in mice ZHAO Hongyu, HU Xiao, SHEN Haitao, ZHENG Qiang 2016, 44 (6):  704-707.  doi: 10.11958/20150188 Abstract ( 756 )   PDF (921KB) ( 3973 )   Abstract: Objective To study the relationship between receptor interacting protein (RIP)1 and hepatocyte necroptosis in isoniazid (INH) induced mouse model. Methods Kunming male mice were randomly divided into three groups. Control group (C) received 0.3 mL of normal saline one time per day. INH group (INH) was injected intraperitoneally INH 100 mg/kg body weight, one time per day. Nec-1 + INH group was injected intraperitoneally Nec-1 in 0.1% DMSO and 1 mg/kg body weight one time/12 hours, and INH was injected intraperitoneally at the same dose with that of INH group. All animals were treated for 7 days. Pathological changes of liver tissues were studied by HE staining. RIP1 expression was detected by immunohistochemical, Western blot and real-time PCR analysis. Levels of malondialdehyde (MDA), reactive oxygen species(ROS), glutathione (GSH) and superoxide dismutase (SOD) in liver homogenate were determined by colorimetric method. Results Hepatocytes were arranged orderly in C group. The degeneration and necrosis of hepatocytes were found in Nec-1+INH group, and severe degeneration and necrosis of hepatocytes were found in INH group. Compared with C group, the expression levels of RIP1, ROS and MDA were increased significantly, and the expression levels of GSH and SOD were decreased significantly in INH group (P＜0.05). INH-induced acute liver necroptosis was significantly alleviated after treatment with Nec-1. Compared with INH group, the expression levels of RIP1, MDA and ROS were significantly decreased, and the expression levels of GSH and SOD were significantly increased in Nec-1+INH group (P＜0.05). Conclusion These results suggest that RIP1 is involved in INH-induced hepatocyte necroptosis in mice. The inhibition of RIP1 expression might be a treatment strategy for prohibition of INH-induced acute liver necroptosis. Related Articles | Metrics

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Effects of the synthetic peptide coatings inhibiting the growth of Streptococcus gordonii on titanium surface MA Qing, ZHANG Xi, ZHANG Xu, SUN Yingchun, GAO Ping 2016, 44 (6):  708-711.  doi: 10.11958/20150285 Abstract ( 904 )   PDF (1046KB) ( 3743 )   Abstract: Objective To detect the independently designed synthetic peptide adsorbed to the titanium surface and its inhibitory effect on streptococcus gordonii, and to provide a new means for antibiosis reseach on oral implants. Methods The physical and chemical properties of the synthetic peptide and antimicrobial peptide were measured by ExPASy Prot⁃Param tool, ProtScale analysis, circular dichroism and Zeta potential instrument. The synthetic peptide was anchored on the surface of the titanium specimen through incubation at room temperature. The adsorption of the synthetic peptide to the titanium surface was examined by X-ray photoelectron spectroscopy (XPS) and the atomic force microscope (AFM). The inhibitory effect on streptococcus gordonii of the synthetic peptide fixed on the titanium surface was viewed by confocal laser scanning microscopy (CLSM). The destructive effects of the synthetic peptide and the antimicrobial peptide on streptococcus gordonii were observed through the transmission electron microscope (TEM). Results The independently designed synthetic peptide still had the physical and chemical properties that the antimicrobial peptide desired. The synthetic peptide had already been detected on the titanium surface after incubated in a 5 g/L synthetic peptide solution. The titanium specimen fixed with the synthetic peptide inhibited the survival and adhesion of streptococcus gordonii. Conclusion It suggests that the independently designed synthetic peptide might have reached the goal of bacterial inhibition on the titanium surface. Related Articles | Metrics

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The role of Foxp3+T cells in chronic intermittent hypoxia induced liver injury BAI Xiaochun1, WANG Lin1, ZHAO Xian2, TIAN Jianli1 2016, 44 (6):  712-715.  doi: 10.11958/20150217 Abstract ( 932 )   PDF (342KB) ( 3832 )   Abstract: Objective To explore the role of Foxp3 + T cells (Tregs) in liver injury induced by chronic intermittent hypoxia. Methods Thirty-two male Wister rats were divided into four groups: control group (A), high-fat diet group (B), intermittent hypoxia group (C), and high- fat diet and intermittent hypoxia group (D). After 4 weeks, blood samples were collected and livers were surgically removed. Using the standard automatic clinical analyzer to test serum levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL- C), alanina aminotransferase (ALT) and aspartato aminotransferase (AST). The MDA content of liver tissue was measured by colorimetrc method. The levels of TNF-α and IL- 1β were measured by radiommunoassay, and the expression of Foxp3 protein was measured by Western blotting technique. Results Serum levels of TC and LDL-C were significantly higher in B group than those of A, C and D groups, and which were higher in D group than those of A and C groups (P＜0.05). There were no significant differences in serum levels of TC and LDL-C between A group and C group. Serum levels of ALT, AST, MDA, TNF-α and IL-1β were significantly higher in C group than those of A group, and which were significantly higher in D group than those of A, B and C groups (P＜0.05). There were no significant differences in these indicators between A group and B group, and between B group and C group. Foxp3 protein expression in liver was significantly lower in D group than that of other groups (P＜0.05). Conclusion Foxp3+ T regulatory cells involve in the regulation of hepatic injury induced by chronic intermittent hypoxia on the basis of a high-fat diet, and which may play an important role in this process of protective immune response. Related Articles | Metrics

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Effects of celecoxib on expression of protein Apaf-1 and function of mobility after severe craniocerebral injury in rats ZHANG Tao1, GUO Jianfei1, XING Linlin1, ZHANG Jinling2, ZHANG Yuxin3 2016, 44 (6):  716-719.  doi: 10.11958/20150231 Abstract ( 960 )   PDF (1205KB) ( 4158 )   Abstract： Objective To investigate effects of celecoxib on the expression of cyclooxygenase- 2 (COX-2), apoptotic protease activation factor-1 (Apaf-1) and function of mobility in rat model of severe craniocerebral trauma. Methods For⁃ ty-eight adult male Wistar rats were randomly divided by random number table into four groups. Normal group was given no manipulation. Sham group was given scalp incision and sutured. The severe closed craniocerebral injury model was estab⁃ lished via Foda method in rats of injury group. Treatment group was given intraperitoneal injection of celecoxib [250 mg/(kg·6 h)] on the basis of injury group. The intraperitoneal injection of same volume of normal saline was given in the other three groups. Samples were taken altogether after 72 hours. Changes of COX-2 and Apaf-1 were detected by immunohistochemis⁃ try and Western blot assay. Ten days after the restoration, six rats were taken from each group for assessing neurological im⁃ pairment scale (NSS). Results The expression levels of COX-2 and Apaf-1 were significantly higher in injury group than those of other groups. The expression levels of COX-2 and Apaf-1 were significantly lower in treatment group than those of injury group but the levels were significantly higher than those of sham group and normal group (P < 0.05). NSS scores showed that rats in treatment group improved mobility compared with that of injury group (P < 0.05), but there was difference compared with Sham group and control group (P < 0.05). Conclusion Celecoxib, with its specific inhibitoty effect on pro⁃ tein COX-2, can effectively reduce inflammatory reactions lower the expression of Apaf-1 and reduce apoptosis of neurons, improving the prognosis of dysfunction of mobility after craniocerebral injury. Related Articles | Metrics

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Risk factors and prognosis for carotid artery stenosis in patients undergoing off-pump coronary artery bypass grafting WANG Chengmeng， WANG Zanxin， WEI Minxin 2016, 44 (6):  720-723.  doi: 10.11958/20160024 Abstract ( 955 )   PDF (372KB) ( 4456 )   Abstract： Objective To analyze the risk factors and prognosis of carotid artery stenosis (CAS) in patients who under⁃ went off-pump coronary artery bypass grafting (OPCAB). Methods A total of 342 patients scheduled for OPCAB between June 2013 and June 2015 were included in this study. According to results of preoperative duplex ultrasound examination of carotid arteries, patients were divided into CAS group (≥50% stenosis) and control group (<50% stenosis or no stenosis). The risk factors of CAS, death rate and the postoperative complications were compared between two groups. Results There were significant differences in age, smoking history, hypertension and chronic obstructive pulmonary disease (COPD) between two groups (P < 0.05). Multivariate logistic regression analysis showed that advanced age (OR=1.050, 95%CI:1.014-1.086, P < 0.01), hypertension (OR=2.566, 95%CI:1.299-5.071, P < 0.01) and COPD (OR=7.573, 95%CI:1.106-51.834, P < 0.05) were independent risk factors for CAS in OPCAB surgery. CAS (OR=4.530, 95%CI:1.361-15.078, P < 0.05) and the history of percutaneous coronary intervention (OR=7.685, 95%CI: 2.289-25.800, P＜0.01) were independent risk factors for postop⁃ erative death. Conclusion Risk factors for CAS in patients undergoing OPCAB include advanced age, hypertension and COPD. There is higher risk of death in CAS patients with OPCAB. Related Articles | Metrics

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The infection status of hrHPV and its effect and significance on expression of Caspase-1 and IL-1β in human cervical carcinoma LUO Yuancai1, WANG Baochen2， GUO Lu1, LI Jing1 2016, 44 (6):  724-728.  doi: 10.11958/20150224 Abstract ( 1129 )   PDF (572KB) ( 4715 )   Abstract: Objective To explore the infection status of high risk human papilloma virus (hrHPV) and its effects and clinical significance on expression of cysteinyl aspartate specific protease 1 (Caspase- 1) and interleukin 1 β（IL-1β）in tissues of human cervical carcinoma. Methods A total of 102 patients with cervical carcinoma (cervical carcinoma group), 60 patients with cervical intraepithelial neoplasia (CIN group) and 30 patients with normal cervix (control group) were usedas the research objects. PCR reverse dot hybridization method combined with DNA chip technique were used to detect hrHPV. The expressions of Caspase- 1 and IL-1β were detected by immunohistochemical technique. Data were analyzed between hrHPV positive group and hrHPV negative group, between single type of hrHPV infection group and multiple type of hrHPV infection group. The relationship between caspase-1 and IL-1β expression and clinicopathological parameters in cervical carcinoma patients were observed. Results HrHPV infection was detected in 75 cases（73.5％）in cervical carcinoma group and 11 types of hrHPV were detected. In these 11 cases, single type and multiple type of hrHPV infection were 61 cases（81.3％）and 14 cases（18.7％ ) separately. HrHPV infection rate was much higher in cervical carcinoma group than those in CIN group and control group（36.7％ and 6.7％). Caspase-1 and IL-1β positive rates were significantly higher in cervical carcinoma group（61.8% and 51.0%） than those in control group（26.7% and 23.3%). The positive rate of Caspase-1 was significantly higher in cervical carcinoma group than that in CIN group（40.0%, all P＜0.01). The positive rates of Caspase-1 and IL-1β（77.8% and 74.1%）were higher in hrHPV DNA negative group than those in hrHPV DNA positive group（56.0% and 42.7%). There were no statistical differences in positive rates of Caspase-1 and IL-1β between single type of hrHPV infection group and multiple type of hrHPV infection group（P＞0.05). The difference of positive expressions of Caspase-1 and IL-1β were significantly related with cell differentiation, tumor size, lymphatic metastasis and clinical stage（P＜0.05 or P＜0.01). Conclusion There are single and multiple types of hrHPV infection in cervical carcinoma and the infection rate is high. HrHPVs may promote the progression of cervical carcinoma by restraining the expressions of Caspase-1 and IL-1β. Related Articles | Metrics

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The predictive value of serum detection of HE4 and CA125 in patients with high risk of stageⅠ endometrial cancer CHEN Anxia, TENG Fei, WANG Yingmei, TIAN Wenyan, GAO Jinping, XUE Fengxia 2016, 44 (6):  729-732.  doi: 10.11958/20160374 Abstract ( 962 )   PDF (320KB) ( 3932 )   Abstract: Objective To investigate the predictive value of preoperative serum human epididymis protein 4 (HE4) and CA125 in patients with high risk of stage Ⅰ endometrial cancer（EC）and to identify the optimal cutoff values. Methods Clinical and pathological data of 231 patients with stage Ⅰ EC were included in this study. Patients were divided into high risk group (n=96) and low risk group (n=135). The preoperative serum levels of HE4 and CA125 were measured, and their correlations with clinical pathological features were analyzed. The ROC curves were generated to determine optimal cutoff values of HE4 and CA125 levels with the maximum Youden index for prediction of high risk EC. Results There were significant differences in serum levels of HE4 and CA125 between patients with different depths of myometrial invasion, with or without vascular invasion, with or without lower uterine segment involvement, with different diameters of tumor and different risk classifications of stage Ⅰ EC (P < 0.05). There were significant differences in serum levels of HE4 between patients with different menopausal status, hypertension, pathological types, histological grading and the involvement of cervical endometrial glands (P < 0.05). The preoperative serum levels of HE4 and CA125 were positive correlated (r=0.262, P < 0.05). The AUC value of HE4 for diagnosing stage Ⅰ EC was 0.794（95%CI：0.734-0.854）， the cutoff value was 74 pmol/L, the sensitivity was 75.0%, specificity was 83.0%, positive predictive value was 75.8%, negative predictive value was 82.4% and the accuracy rate was 79.7%. The AUC value of CA125 for diagnosing stage Ⅰ EC was 0.696 (95% CI: 0.624- 0.767), the cutoff value was 17 kU/L, the sensitivity was 56.3% , specificity was 85.9% , positive predictive value was 74.0%, negative predictive value was 73.4% and the accuracy rate was 73.6%, respectively. The AUC value of combination of both markers was 0.847 (95% CI: 0.796- 0.899), the corresponding values were 95.8% , 77.0% , 74.8%, 96.3% and 84.8%, respectively. Conclusion The best cutoff values of HE4 and CA125 are 74 pmol/L and 17 kU/L for detecting high risk stageⅠEC. The combined detection is better than that of the single detection in sensitivity, negative predictive value and accuracy rate. Related Articles | Metrics

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Tankyrase expression in lung bronchiolo-alveolar adenocarcinoma and its relationship with the WNT pathway LI Chong1,2， ZHENG Xu2， HAN Yanyan2， LYU Yan2， LAN Fu2， ZHAO Jie2 2016, 44 (6):  733-735.  doi: 10.11958/20150339 Abstract ( 1317 )   PDF (1171KB) ( 4014 )   Abstract：Objective To explore the expression of tankyrase (TNKS) and its relationship with WNT/β-catenin signal⁃ ing pathway in lung acinar adenocarcinoma. Methods Seventy-two samples of single subtype alveolar like lung adenocarci⁃ noma (lung adenocarcinoma group) and 67 specimens of normal lung tissue adjacent to carcinoma (adjacent to carcinoma group) were collected. Immunohistochemical method was used to detect expressions of TNKS, beta-catenin (β-catenin) and c-myc protein. The correlation of each protein expression in lung adenocarcinoma tissues was analyzed. The differential ex⁃ pression of TNKS was detected by Western blot assay in two groups. Results Tankyrase protein was mainly expressed in cy⁃ toplasm. The expression of β-catenin protein was mainly in cytoplasm and nuclear of lung adenocarcinoma. The expression of β-catenin was mainly in cytoplasm, and a small amount was in nuclear of the adjacent group. The c-myc protein was ex⁃ pressed mainly in the nucleus. The positive expression rates of TNKS, β-catenin and c-myc protein were significantly high⁃ er in lung adenocarcinoma group than those of adjacent to carcinoma group (P＜0.05). The expression of β-catenin in cyto⁃ plasm and nucleus was positively correlated with the expression of TNKS and c- myc (P＜0.05). Western blot analysis showed that the relative expression level of TNKS was significantly higher in lung adenocarcinoma group than that of adja⁃ cent to carcinoma group (0.497±0.021 vs. 0.237±0.015, t=13.00, P＜0.01). Conclusion Abnormally high expression of TNKS in lung adenocarcinoma may promote the occurrence of lung cancer by regulating the WNT signaling pathways. Inhib⁃ iting TNKS expression may become a new target to treat lung adenocarcinoma. Related Articles | Metrics

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Changes and its clinical significance of the plasma growth differentiation factor-15 in patients with chronic congestive heart failure MA Ping1, XU Huan2, XU Yehua2, XU Qingbin1, XIONG Aiqin2 2016, 44 (6):  736-739.  doi: 10.11958/58776 Abstract ( 1066 )   PDF (316KB) ( 4075 )   Abstract: Objective To investigate the changes and clinical significance of the plasma growth differentiation factor- 15 (GDF-15) in patients with chronic congestive heart failure (CHF). Methods A total of 100 patients with CHF were in⁃ cluded in this study (CHF group), and 30 healthy persons were used as control group. CHF group was divided into heart func⁃tionⅡgrade (n=35)， Ⅲ grade (n=32)， Ⅳ grade (n=33) groups in accordance with New York Heart Association (NYHA). And CHF group was also divided into left ventricular ejection fraction (LVEF) <0.4 grade (n=52) and LVEF≥0.4 grade (n=48) groups in accordance with LVEF of patients. The plasma GDF-15 and brain natriuretic peptide (BNP) levels were detected by ELISA. The values of LVEF, left ventricular end-diastolic diameter (LVDd), left ventricular systolic diameter (LVDs) and left ventricular fractional shortening (LVFS) were detected by echocardiography. The correlation of GDF-15, NYHA classifi⁃ cation, BNP and index of echocardiography was analyzed between groups. Results Compared with control group, the levels of BNP, GDF-15, LVDd and LVDs were significantly higher in heart failure group, and values of LVEF and LVFS were sig⁃ nificantly lower (P < 0.05). The plasma levels of BNP, GDF-15, LVDd and LVDs were in turn increased in control group, LVEF≥0.4 grade group and LVEF<0.4 grade group. The plasma levels of LVFS were in turn decreased, in control group, LVEF≥0.4 grade group and LVEF<0.4 grade group (P＜0.05). There were positive correlations between the plasma levels of GDF-15 and BNP, NYHA, LVDd and LVDs (r=0.524, 0.286, 0.453 and 0.531, P < 0.05). The plasma level of GDF-15 was negatively correlated with LVEF and LVFS (r= -0.592, -0.587， P < 0.05). Conclusion The plasma level of GDF-15 can be used as a new marker for diagnosis, treatment and prognosis in patients with chronic congestive heart failure. Related Articles | Metrics

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Study on the predictive value of preoperative magnetic resonance imaging for biochemical recurrence after radical prostatectomy YAN Xuxin1, ZHANG Zhihong2, LIU Xuejun1, HAN Dongmin1, LI Huan1, MA Li1, LI Xiaodong1 2016, 44 (6):  740-743.  doi: 10.11958/20150398 Abstract ( 1156 )   PDF (381KB) ( 5764 )   Abstract： Objective To evaluate the suitability of preoperative magnetic resonance imaging (MRI) positivity as a pre⁃ dictor of biochemical recurrence after radical prostatectomy (RP). Methods Data of 115 diagnosed prostate cancer patients with RP from 2006 to 2013 were collected retrospectively. According to MRI results, patients were divided into MRI positive group (n=87) and MRI negative group (n=28). The preoperative clinical data were compared between two groups. Biochemi⁃ cal recurrence was defined as increase or persistence of serum prostate specific antigen (PSA) levels>0.2 μg/L after surgery. Cox proportional hazard regression model was used to analyze the risk factors associated with biochemical recurrence. Re⁃ sults The survival time was shorter in MRI positive group than that of MRI negative group. The preoperative PSA level was higher in MRI positive group than that of MRI negative group (P < 0.05). There was no significant difference in the age of pa⁃ tients between two groups. In MRI positive group, the proportion of preoperative PSA>20 μg/L , the proportion of patients at clinical stage T3, the proportion of patients with pathological Gleason score≥8, the proportion of patients with seminal vesi⁃ cle invasion (SVI) and the proportion of patients with endocrine therapy were higher than those of negative group (P < 0.05). The univariate analysis showed that preoperative MRI positivity, PSA level, biopsy Gleason score, clinical stage, pathological Gleason score, SVI level and lymph node invasion (LNM) were associated with biochemical recurrence (P < 0.05)．However, the multivariate analysis showed that only preoperative MRI positivity, biopsy Gleason score and clinical stage were indepen⁃ dent prognostic factors (P < 0.05). Conclusion Preoperative MRI positivity can predict biochemical recurrence after RP successfully, and improve the prognosis of patients. Related Articles | Metrics

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Analysis of risk factors for multiple drug resistant bacterial infections in chronic obstructive pulmonary disease XIE Zhaoyun1,2, XIONG Yun1, SUN Jing1, HU Yang2, LI Yaofu1 2016, 44 (6):  744-747.  doi: 10.11958/20150111 Abstract ( 1040 )   PDF (301KB) ( 3849 )   Abstract: Objective To analyze the risk factors of multiple drug resistant bacterial infections in patients with chronic obstructive pulmonary disease (COPD), and provide guidance for disease control and prevention. Methods Clinical data of 814 COPD patients were retrospectively analyzed from June 2011 to May 2015, including patient's age, gender, smoking history, age of onset, severity, aggravated frequency, duration of exacerbations, diabetes mellitus, complications, use frequency and use duration of glucocorticoid, use frequency of antimicrobial agents and use duration of each time, types of antimicrobial drugs used, combined with antibacterial drugs, plasma albumin concentration, blood glucose, bacteria culture detection of multi drug resistant bacteria infection. The risk factors of multi drug resistant bacteria infection were analyzed. Results A total of 857 pathogenic bacteria were isolated from 814 COPD patients with pulmonary infection. Multiple drug resistant bacteria infection were detected in 170 cases, and 175 strains (20.42%) were detected. The detection rate of multi drug resistant/PAN resistant pseudomonas aeruginosa (MDR/PDR- PA) was 55.38% (36/65). There were significant differences in patients with multi drug resistant bacteria infection between different clinical pathological characteristics. Logistic regression analysis showed that the acute exacerbation duration (days), long time use of antimicrobial drugs, and high frequency of corticosteroids and antibiotics use were independent risk factor of multi drug resistant bacteria infection in COPD patients. Conclusion Prevention and treatment of multiple drug resistant bacteria infection in COPD patients should pay attention to the combination of community and hospital, and take effective measures to prevent and control the risk factors. Related Articles | Metrics

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Clinical diagnosis and treatment of high-risk small renal cell carcinoma WU Yudong1， LI Gang1， WANG Jiang1,2， WANG Yong1， NIU Yuanjie1 2016, 44 (6):  748-751.  doi: 10.11958/20160127 Abstract ( 971 )   PDF (1530KB) ( 4325 )   Abstract: Objective To investigate the high-risk factors leading to a poor prognosis of small renal cell carcinoma, and provide theoretical basis for the individualized treatment regimen. Methods This retrospective study analyzed the clini⁃ cal and histological data of 18 patients with small renal cell carcinoma treated in the Department of Urology of the Second Hospital of Tianjin Medical University from January 2004 to July 2015. All the patients underwent ultrasound, plain and en⁃ hanced CT examinations, also, received the surgeries. The tumor diameters, pathological types, pathological stages, Fuhrman grading of tumors and the prognosis of patients were analyzed. Results Preoperative CT examination revealed that 18 pa⁃ tients with the average tumor diameter of (3.1 ± 0.6) cm (ranged 2.0 to 4.0 cm). Five patients were diagnosed as T1aN0M0, 4 patients with T1aN0M1 (3 cases with lung metastasis, 1 case with brain metastasis), 3 patients with T1aN1M0 (CT examina⁃ tion showed a lymph node metastasis), 6 patients with T3aN0M0 (renal vein invasion or renal vein tumor thrombus). Patholog⁃ ical examination after surgery showed that 12 patients were Fuhrman grade Ⅱ, 5 were grade Ⅲ and 1 was grade Ⅳ;15 cases were clear cell carcinomas, 1 case was papillary carcinoma, 1 was hybrid cellular tumor (malignant rhabdoid tumor with sar⁃ comatoid differentiation) and the last case was sarcomatoid carcinoma renal cell carcinoma (Fuhrman grade V). 4 patients (T3a, Fuhrman grade Ⅱ) underwent retroperitoneal laparoscopic partial nephrectomy and the remaining underwent laparo⁃ scopic radical nephrectomy. The median follow-up time was 22.5 months (ranged 6 to 48 months). Four cases died (2 cases with tumor diameters of 3.8 cm and 4.0 cm at preliminary diagnosis， 2 cases with sarcomatoid renal carcinoma and 1 with brain metastasis), 1 case was lost. Other patients were found no tumor recurrence and metastasis. Conclusion The small re⁃ nal cell tumor with diameter≥3.0 cm, Fuhrman Ⅲ/Ⅳ grade， sarcomatoid cancer or metastasis should be considered as highrisk factors of small renal cell carcinoma. The high-risk small renal cell carcinoma is heterogeneous in its biological behav⁃ ior, which is expressed as aggressive growth and early invasion of renal tissue and even metastasis. The individualized treatment should be made based on preoperative imaging findings and postoperative pathology. Related Articles | Metrics

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The predictive values of HIF-1 and VEGF in early diabetic nephropathy SONG Qiuyan, YU Ling, DONG Ruihong 2016, 44 (6):  752-754.  doi: 10.11958/20150092 Abstract ( 883 )   PDF (325KB) ( 4159 )   Abstract：Objective To investigate the predictive values of hypoxia- induced factor(HIF)- 1α and vascular endothelial growth factor (VEGF) in patients with type 2 diabetic kidney disease, and its relationship between HIF- 1α, VEGF and glomerular filtration rate (eGFR). Methods Seventy- six patients with type 2 diabetic kidney disease were divided into two groups: DKD1 group [eGFR＞90 mL/(min·1.73 m2)， n=48] and DKD2 group [60 mL/(min·1.73 m2)＜eGFR≤ 90 mL/(min·1.73 m2), n=28], according to eGFR results. Twenty-three cases with healthy physical examination were used as the control group. The liquid phase immunoprecipitation and transmission turbidity methods were used to detect 24-h urine albumin, and urinary albumin excretion rate (UAE) was calculated. The serum VEGF and HIF-1α levels were measured by ELISA. Results Serum levels of UAE, HIF-1α and VEGF were significantly higher in DKD1 and DKD2 groups than those in control group (P < 0.05). There was no significant difference in UAE levels between DKD1 and DKD2 groups (P＞0.05). Serum levels of HIF-1α and VEGF were significantly higher in DKD2 group than those of DKD1 group (P＜0.05). There were negative correlation between VEGF and HIF-1α levels with eGFR level (r=-0.59, P < 0.01; r=-0.55, P < 0.01). ROC curve showed that serum VEGF and HIF-1α levels were above the opportunity diagonal. The diagnostic sensitivities of HIF- 1 to DKD1 and DKD2 were 92.1% and 87.4%. The diagnostic specificities of HIF-1 to DKD1 and DKD2 were 91.7% and 85.4%. The diagnostic sensitivities of VEGF to DKD1 and DKD2 were 96.1% and 86.2%, and specificities were 93.3% and 89.1%. Conclusion Serum levels of VEGF and HIF-1α change with the severity of diabetic nephropathy are likely to be an early predictor for the progression of diabetic kidney disease. Related Articles | Metrics

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The expression and clinical significance of α-actinin-1 in prostate cancer and benign prostatic hyperplasia LI Yapeng1, LIU Jian1, CAO Fenghong1, LI Xiaoqiang1, CHEN Ning1,2, CHEN Chen1,2 2016, 44 (6):  755-758.  doi: 10.11958/20150262 Abstract ( 1081 )   PDF (423KB) ( 5466 )   Abstract：Objective To investigate the expression and clinical significance of α- actinin- 1 protein (ACTN1) in prostate cancer (PCa) and prostatic hyperplasia (BPH). Methods The clinical data of patients with PCa or BPH treated in our school affiliated hospital were collected between January 2007—October 2014, according to certain criteria. Immunohistochemistry method was used to detect the expression of ACTN1 in 30 samples of PCa and 30 samples of BPH tissues. Western blot assay was used to detect the relative expression of ACTN1 in 18 samples of PCa and 20 samples of BPH tissues in two groups. Results The result of immunohistochemistry showed that the positive expression rates of ACTN1 were 76.7% and 20% in PCa and BPH groups respectively. The difference was statistically significant (P < 0.01). Western blot assay showed that the relative expression of ACTN1was significantly higher in PCa group (0.591±0.182) than that in BPH group (0.037 ± 0.052, P < 0.05). There was no significant difference in expression level of ACTN1 betweendifferent age groups. There was no significant difference in serum prostate specific antigen (PSA) level between patients with or without bone metastasis, and patients with or without lymph node metastasis (P > 0.05). There were significant differences in ACTN1 levels between different Gleason score and T staging groups (P < 0.05). Conclusion The expression of α-actinin-1 is significantly higher in PCa tissues than that in BPH tissues. There is the relationship between expression of ACTN1, Gleason scores and T staging. Related Articles | Metrics

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Analysis of clinical characteristics and perinatal outcomes in pre-pregnancy obesity patients with early-onset preeclampsia WU Fan， HUA Shaofang， LI Yueqin 2016, 44 (6):  759-762.  doi: 10.11958/20150320 Abstract ( 791 )   PDF (355KB) ( 3849 )   Abstract: Objective To explore clinical characteristics and the perinatal effects of multiple systems in prepregnancy obesity patients with early-onset preeclampsia. Methods A total of 111 cases with early-onset preeclampsia in our Inpatient Obstetric Department were reviewed in this study. According to pre- pregnancy body mass index (BMI), patients were divided into normal before pregnancy group (BMI＜28 kg/m2, n=56) and obese before pregnancy group (BMI≥ 28 kg/m2, n=55) . The body weight, BMI, weight gain during pregnancy, risk factors, clinical features, and clinical features of neonatal index were compared between two groups. The correlation between maternal and neonatal indexes and glucose and lipid indexes were compared. Results Obese before pregnancy was more prone to blood concentration and disorders of lipid metabolism. The serum levels of lipid, glycosylated hemoglobin (HbA1c), hematocrit, platelet and fibrinogen were significantly higher in obese before pregnancy group than than those of normal group. The induced neonatal acidosis and other organ damage, newborn Apgar score, pH, glucose were significantly lower in obese before pregnancy group than those of normal group. The residual alkali (BE), lactic acid (LAC), creatine kinase (CK) and creatine kinase (CKMB) were higher in obese before pregnancy group than those of normal before pregnancy group. Correlation analysis showed that maternal low density lipoprotein cholesterol (LDL- C), total cholesterol (TC), three acyl glycerol (TG) and HbA1c were negatively correlated with neonatal Apgar score and pH, and which were positively correlated with neonatal LAC and CKMB. Both differences were significant (P＜0.05 or P＜0.01). Conclusion The dyslipidaemia in early onset preeclampsia in obesity patients before pregnancy is closely related with maternal and perinatal outcomes. Related Articles | Metrics

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To analyze the risk factors of tetany in patients with hyperventilation syndrome YOU Daofeng1, QIAO Qiuge2, XIE Bing1, JIANG Xiangming1, CHEN Ying1 2016, 44 (6):  763-765.  doi: 10.11958/20150230 Abstract ( 1064 )   PDF (285KB) ( 3747 )   Abstract： Objective To explore the risk factors causing tetany in patients with hyperventilation syndrome. Meth⁃ ods A total of 103 patients with hyperventilation syndrome treated in our hospital were included in this study. According to whether there was tetany, patients were divided into tetany group and non-tetany group. Values of gender, age, electrolyte, pH and p(CO2) were analysed between two groups. The factors of P < 0.1 were engaged in binary Logistic regression. Logistic regression (Forward Wald) was used to analyze the risk factors of tetany in patients with hyperventilation syndrome. Re⁃ sults In 103 patients there were 70 patients with tetany (68%), 33 patients without tetany(32%). The serum K+, serum phos⁃ phorus and p(CO2) values were significantly lower in tetany group than those of non-tetany group (P < 0.01), while the pH value was significantly higher in tetany group than those of non-tetany group (P < 0.01). There were no significant differences in gen⁃ der, age, serum Na+, serum Cl-, serum calcium (bound calcium and ionized calcium), ionized calcium and serum Mg2+ levels be⁃ tween two groups (P > 0.05). It was revealed that the younger age, the lower level of the serum K+, serum phosphorus and p(CO2) were the risk factors of tetany through binary Logistic regression analysis. Conclusion The risk factors of tetany in patients with hyperventilation syndrome include younger age, lower level of serum K+ and serum phosphorus and reduced p(CO2). Related Articles | Metrics

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Vitreopapillary traction： a possible reason of anterior ischemic optic neuropathy MENG Bin， CHEN Xu， JI Ang 2016, 44 (6):  766-768.  doi: 10.11958/20160008 Abstract ( 984 )   PDF (519KB) ( 3996 )   Abstract： Objective To discuss the relevance between vitreopapillary traction (VPT) and anterior ischemic optic neu⁃ ropathy (AION). Methods Two patients suffering from AION were underwent routine ophthalmic examination, and visual field (VF), fundus fluorescence angiography (FFA) and optical coherence tomography (OCT) examination. The images were analyzed. Results VPT syndrome was observed by OCT in both cases. In addition, the affected parts of VPT were consis⁃ tent to that of AION. Conclusion Vitreopapillary traction is a possible reason of anterior ischemic optic neuropathy. Related Articles | Metrics

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Effects of serum level of TPOAb on Graves disease in patients using anti-thyroid drug treatment LU Zefen1, YU Jia2, REN Liping3, HAO Jie1, HE Qing4 2016, 44 (6):  769-771.  doi: 10.11958/20150409 Abstract ( 1002 )   PDF (259KB) ( 4036 )   Abstract: Objective To analyse effects of the serum levels of thyroid peroxidase antibodies (TPOAb) on antithyroid drugs (ATD) treatment in patients with incipient Graves disease (GD). Methods A total of 121 patients with incipient GD, who were used anti thyroid drugs for 12 months, were included in this study. Patients were dvided into two groups: TPOAb negative group (TPOAb≤35 IU/mL, n=49) and TPOAb positive group (TPOAb > 35 IU/mL, n=72). According to the degree of TPOAb drops the TPOAb positive group was sub-divided into low level positive group (35 IU/mL < TPOAb≤200 IU/mL, n=19), medium level positive group (200 IU/mL＜TPOAb≤ 500 IU/mL, n=20), high level positive group (500 IU/mL＜ TPOAb≤1 000 IU/mL, n=13), and very high level positive group (TPOAb > 1 000 IU/mL, n=20). The ATD total dosage for 12 months and thyroid-stimulating hormone (TSH), which returned to normal rate after treatment of 3, 6 and 12 months, were compared between groups. Results ATD total dose was lower in TPOAb positive group (1 743.82±265.38) mg than that of negative group (1 889.18 ±125.51) mg. The ATD total dosages were (1 759.71±230.29) mg, (1 793.75±299.02) mg, (1 731.54± 236.44) mg and (1 710.00±290.73) mg for low level TPOAb positive group, medium level TPOAb positive group, high level TPOAb positive group, and very high level TPOAb positive group respectively. The recovering ratio of TSH was significantly higher in TPOAb positive group than that of TPOAb negative group after 3-month treatment (P < 0.05). Conclusion TPOAb positive serum can lead to shorten the course of ATD treatment and reduce the total amount of ATD treatment in GD patients. Related Articles | Metrics

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A comparative study of laparoscopic neoplasty and open neoplasty on levels of procalcitonin and cell-mediated immunity in elderly patients with upper digestive tract ulcer perforation WANG Gengzhuang1, ZHAO Zhirong2, PAN Lifeng1 2016, 44 (6):  772-775.  doi: 10.11958/20150273 Abstract ( 1029 )   PDF (314KB) ( 3753 )   Abstract：Objective To study the effect of laparoscopic neoplasty and changes of procalcitonin (PCT) and cellmediated immunity in elderly patients with duodenal ulcer perforation. Methods Forty-four elderly patients with duodenal ulcer perforation were divided into laparoscopic neoplasty group (n=20) and open neoplasty group (n=24)．The level of PCT was assayed by colloid immunization. Blood levels of CD3+, CD4+, CD8+ and natural killer (NK) cells were detected by flow cytometry before operation and at 0.5 h, 24 h, 48 h and the 7th day after operation．Surgical complications and postoperative hospital stay were observed in two groups. Results There was no significant difference in the activity of PCT before operation between two groups. There were no significant differences in CD3 + , CD4 + , CD8 + and NK cells 0.5 h before operation between two groups. The level of PCT was significantly lower at 24 h and 48 h after operation in laparoscopic group than that in open operation group (P< 0.05). There were significant differences in the serum levels of CD3+, CD4+, CD8+ and NK cells with time changes in two groups (P＜0.05). There were significant decreases in CD3+, CD4+ and CD8+ at 24 h after operation than those before operation in two groups, which were significantly lower in open operation group than those of laparoscopic group except for CD8+. The levels of CD3+, CD4 + and CD8+ recovered to the levels before operation at 48 h after operation in laparoscopic group. In open operation group CD3+ did not return to the level before operation at 7 d after operation. The levels of NK cells were decreased at 24 h after operation in two groups, but there was no significant difference in the level of NK cells between two groups. The level of NK cells returned to preoperative levels at 48 h after operation inlaparoscopic group, while there was no significant difference in the level of NK cells before and after operation in open neoplasty group. Compared with the open neoplasty group, there were fewer complications and shorter hospital stay in laparoscopic group. Conclusion The laparoscopic repair operation for gastric and duodenal ulcer perforation has a fewer effects on the stress response and immune function in elderly patients, and has the advantages in the protection of immune function. Related Articles | Metrics

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The short term outcome after mitral valve replacement with the perimount bovine pericardial valve TIAN Yanchao1, CHEN Qingliang2, CHEN Tongyun2, JIANG Nan2, WANG Lianqun2 2016, 44 (6):  776-779.  doi: 10.11958/20150341 Abstract ( 1067 )   PDF (324KB) ( 4043 )   Abstract: Objective To evaluate the short term outcome after mitral valve replacement with the Perimount bovine pericardial valve. Methods Eighty-eight patients underwent mitral valve replacement with the bovine pericardial valve in hospital were included in this study. Postoperative general condition including mortality and cerebral hemorrhage was observed. The hemodynamic and New York heart disease association (NYHA) heart function classification were recorded by Doppler echocardiograms before operation, postoperative 1 week, 3 months and 1 year after mitral valve replacement. Values of the different time points of NYHA, left atrial diameter (LA), left ventricular end diastolic diameter (LVDD), left ventricular end systolic diameter (LVSD), left ventricular ejection fraction (LVEF) and pulmonary artery pressure (PAP) were compared. The hemodynamic parameters were also compared including the peak cross valve pressure (PG), mean cross valve pressure (MG), peak cross valve velocity (PV) and effective orifice area (EOA) 1 week, 3 months and 1 year after surgery. Results There were two cases (2.3%) dead in one year (one died of cerebral hemorrhage and another one died of thromboembolism). There was 1 perivalvular leakage (1.2%). There was no endocarditis or structural valve deterioration. NYHA cardiac function was improved at postoperative 3 months and 1 year (P < 0.05). The values of LA, LVDD and PAP were significantly decreased at postoperative 1 week, 3 months and 1 year compared with those before operation (P < 0.05). Values of LVSD and LVEF were significantly decreased at postoperative 1 week compared with those before operation (P < 0.05). The value of LVEF was significantly increased at postoperative 3 months and 1 year (P < 0.05). Compared with postoperative 1 week, the values of LA and LVEF were significantly increased at postoperative 3 months and 1 year (P < 0.05). There were no significant differences in PG, MG, PV and EOA between postoperative 1 week, 3 months and 1 year. Conclusion With the excellent performance of cardiac function recovery, left ventricular restoration and hemodynamic, the Perimount bovine pericardial valve remains a reliable choice as a mitral tissue valve. Related Articles | Metrics

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Akin combined Scarf osteotomies for moderate and severe hallux valgus LI Xiaodong, JIANG Han, JIANG Yi, XIAO Lianping, ZHENG Jinxin 2016, 44 (6):  780-782.  doi: 10.11958/20150332 Abstract ( 1403 )   PDF (402KB) ( 4074 )   Abstract: Objective To study the clinical efficacy of Akin combined Scarf osteotomies for moderate and severe hal⁃ lux valgus. Methods Thirty-nine (58 feet) patients received Akin combined Scarf osteotomies, in which 12 patients (16 feet) with moderate hallux valgus received the single scarf osteotomy. Hallux valgus angle (HVA), intermetatarsal angle (IMA), tibial sesamoid position and American orthopedic foot and ankle society score (AOFAS) were measured and compared before and after operation respectively. Results All operations were successfully completed patients were followed up for 8- 22 months. The mean operation time was (55.0±6.8) min. The amount of intraoperative bleeding was 3-20 mL with an aver⁃ age of (11.0±5.4) mL. All patients were healed except for a delayed union of Akin on a severe hallux valgus patient. Two cas⁃ es (2 feet) were found inflammation in surgical incision. Two cases (2 feet) were found numbness in dorsal medial side of hal⁃ lux, which was considered nerve damage and improved in 3-5 months. There was no recurrence of hallux valgus in the peri⁃ od of following up. After surgery, HVA (14.1°±5.3°), IMA (7.7°±3.8°) and tibial sesamoid position (2.58±0.61) were signifi⁃ cantly decreased compared with those before operation (39.6°±6.8°, 18.7°±5.4° and 4.87±0.59, P＜0.05). AOFAS score (84.4±8.7) was significantly higher after surgery than that before surgery (37.3±9.5, P＜0.05). Conclusion Akin combined Scarf osteotomies can achieve a excellent therapeutic effect for moderate and severe hallux valgus with very few complica⁃ tions and recurrence, which is worth for clinical application. Related Articles | Metrics

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Comparison of curative effects of venlafaxine on the depression between the therapeutic modes of inpatient and outpatient REN Liping, LIU Peipei 2016, 44 (6):  783-785.  doi: 10.11958/20150198 Abstract ( 1354 )   PDF (341KB) ( 3809 )   Abstract： Objective To evaluate the curative effects of venlafaxine in patients with depressive disorder between therapeutic modes of inpatient and outpatient. Methods Seventy-two patients with depressive disorder including 36 inpatients and 36outpatients were measured respectively with HAMD24 after treatment for 1 week, 2 weeks, 4 weeks and 8 weeks, and then the curative effects were compared between two groups. Results It was found that all patients in the inpatient group completed the observation and two patients in the outpatient group didn’ t complete the observation. The HAMD24 score was significantly decreased in the inpatient group than that of the outpatient group from the second week to the end of the eighth week (P < 0.05). The total effective rates were 80.6% for inpatient group and 64.7% for outpatient group, no significant difference between them. Conclusion Venlafaxine shows a higher therapeutic value and outcome in the therapeutic mode of inpatient. Related Articles | Metrics

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Analysis of related factors of postoperative delirium in elderly patients SHAO Yize1, ZHANG Yanqiu1, LU Xiangli2 2016, 44 (6):  786-788.  doi: 10.11958/20150208 Abstract ( 902 )   PDF (296KB) ( 3909 )   Abstract：Objective To explore related influencing factors for postoperative delirium in elderly patients, to provide a reference for the prevention and treatment. Methods Fifty-four patients with delirium after surgery were used as observa⁃ tion group, and a total of 150 subjects with no delirium after surgery during the same period were selected as the control group. Data of age, gender, malnutrition, disorders of water and electrolyte metabolism, postoperative mechanical ventilation, postoperative hypoxemia, severe infection and postoperative pain degree, and the combination of basic diseases were com⁃ pared and analysed between two groups. The binary logistic regression analysis was used to analyse the influencing factors of postoperative delirium. The outcome and prognosis were observed and analyzed in observation group. Results The average age was significantly higher in observation group than that of control group (P＜0.05). The percentages of postoperative hy⁃ poxemia and severe infection were significantly higher in observation group than those of control group (P < 0.05). Patients with higher age, postoperative hypoxemia and severe infection were risk factors for postoperative delirium. In observation group,1 case died of lung infection, 1 case died of multiple organ failure, the remaining 52 patients were improved and dis⁃ charged from hospital after three months. Conclusion For patients with higher age, postoperative hypoxemia and severe in⁃ fection are the risk factors for occurrence of postoperative delirium. More attention should be paid to clinical preoperative and postoperative periods. Related Articles | Metrics

Drug Clinical Evaluations

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Comparison of efficacies of levosimendan and recombinant human brain natriuretic peptide in patients with acute decompensated heart failure WU Shubin, DING Liping, JIN Zhitao, ZHANG Lijuan, ZHANG Zheng, KANG Fengchi, ZHANG Lian, HU Taohong 2016, 44 (6):  789-792.  doi: 10.11958/20150272 Abstract ( 1332 )   PDF (351KB) ( 3947 )   Abstract： Objective To compare the effects of levosimendan (Levo) and recombinant human brain natriuretic peptide (rhBNP) in patients with acute decompensated heart failure (ADHF). Methods Seventy-five patients were included into this randomized positive-controlled and parallel-group study to receive either Levo (Levo group), rhBNP (rhBNP group) or dobutamine therapy (control group). Heart rate, respiratory rate, 24-hour urine volume， improvement in six-minute walktest after 72-h treatment were compared between three groups. The blood level of BNP and values of left ventricular end dia⁃ stolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) after one- week treatment were compared between three groups. Results After 72-h treatment, heart rate and respiratory rate were significantly decreased, and 24-hour urine volume, six-minute walk-test were significantly increased in three groups (P < 0.05). The NT-proBNP and LVEF were im⁃ proved after one week treatment (P > 0.05), but the LVEDD was improved barely (P > 0.05). The improvements were more significant in Levo group and rhBNP group compared to those of control group (P < 0.05). There were no significant differenc⁃ es in improvements between Levo group and rhBNP group. The common adverse reactions were hypotension and ventricular premature beats. There were no significant differences in adverse event rates between three groups (P < 0.05). Conclusion As compared with dobutamine, Levo and rhBNP have optimized efficacy, fewer side effects and good safety in the treatment of ADHF. They are worth of clinical application. Related Articles | Metrics

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Clinical observation of the efficacy of No. 1 Baiban granules on vitiligo ZHAN Qingxia, GU Xinping, GU Ankang, ZHANG Junling 2016, 44 (6):  793-795.  doi: 10.11958/20150362 Abstract ( 1374 )   PDF (680KB) ( 3867 )   Abstract: Objective To observe the efficacy of No. 1 Baiban granules in the treatment of vitiligo using skin confocal laser scanning microscopy. Methods The study was approved by the hospital ethics committee. Nine-two patients with qi stagnation and blood stasis were collected from Department of Dermatology of the Affiliated Hospital of Tianjin Academy of Traditional Chinese Medicine and were randomly assigned to two groups. The experimental group (n=49) received oral Baiban granule Ι, 3 g, twice a day, transfer factor capsules 6 mg, 3 times a day, 0.1% mometasone furoate cream (excluding the face) for external use, once a day, or 0.1% tacrolimus ointment (Afacial), twice a day for 3 months. The control group (n= 43) only received oral transfer factor capsules 6 mg, 3 times a day, 0.1% mometasone furoate cream (except for the face) for external use, once a day, or 0.1% tacrolimus ointment (face), twice a day, treatment for 3 months. The therapeutic effect was observed in two groups. CLSM was used to monitor the compound color and pigment recovery level, and changes of the number of dendritic cells and inflammatory cell infiltration in skin. Results The compound color was improved significantly after treatment in experimental group and control group. The total effective rate was higher in experimental group than that of control group (81.63% vs. 69.77%，χ2=3.947，P＜0.05). Results of CLSM observation showed that the pigment recovery of skin lesion, the proportion of dendritic cells and inflammatory cells were significantly higher in experimental group than those of control group (P < 0.05). Conclusion The treatment effect of No. 1 Baiban granules is more obvious for vitiligo of qi stagnation and blood stasis. Related Articles | Metrics

Review

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Research progress of sequencing method for animal mitochondrial genome LI Tianjie1, CAO Yanxiang2, ZHAO Hongcui1,3,4, YU Yang1,3,4, QIAO Jie1,3,4 2016, 44 (6):  796-800.  doi: 10.11958/20150437 Abstract ( 1272 )   PDF (393KB) ( 4435 )   Abstract: Mitochondria, the power house of cells, are important organelles in eukaryotic cells. Having their own unique and complete DNA (mtDNA) and genetic system, mitochondria play an essential role in cellular energy metabolism, intracel⁃ lular signaling and apoptotic pathways, as well as many other biological functions, which are closely related with cellular met⁃ abolic network. A disruption of mitochondrial genes can therefore result in mitochondrial dysfunction and human diseases, thus they have been widely used in molecular biology, development biology, genetics, forensic identification and clinical diag⁃ nosis. Consequently, sequencing mitochondrial genome has shown great significance in mitochondrial structure and function research. In this review, research progress in mitochondrial genome sequencing method is summarized, mainly focusing on Sanger sequencing, long-PCR and next-generation sequencing. Also rolling circle amplification and indirect sequencing of mtDNA are reviewed. The ambiguities caused by numts in indirect sequencing are mentioned and resolved. Related Articles | Metrics