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15 October 2016, Volume 44 Issue 10 Previous Issue    Next Issue
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Cell and Molecular Biology



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Effects of long non-coding RNA HOTAIR on proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo GUO Wenyu, KONG Lingping, SUN Shanshan, WANG Yu, ZHAO Minghui, ZHOU Xuan, WANG Xudong, ZHANG Lun 2016, 44 (10):  1185-1189.  doi: 10.11958/20150403 Abstract ( 855 )   PDF (1349KB) ( 3998 )   Objective To investigate the influence of long non-coding RNA HOTAIR in proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo. Methods siHOTAIR was used to inhibit the HOTAIR expression in Tb3.1 human tongue squamous cell carcinoma cell line. The experiments were divided into siHOTAIR group, nonsense sequence group and blank control group. Real-time PCR was used to detect the HOTAIR expression. MTT assay was employed to determine the cell survival. The expression levels of Bcl2, BAX, caspase- 3, cleaved caspase- 3 were examined by Western blot assay. Tb3.1 xenograft tumor model was established in BALB/c nude mice, and the tumor model was divided into control group, negative group, and siHOTAIR treated group. The tumor tissues were measured by immunohistochemistry stain (IHC) and TUNEL assay. Results The detection of real- time PCR showed that HOTAIR expression was reduced after treated with siHOTAIR. Western blots assay showed that Bcl-2 protein was suppressed while cleaved caspase- 3 and BAX protein were up-regulated after treated with siHOTAIR. MTT assay indicated that the cell survival rate was significantly reduced in siHOTAIR treated group. Flow cytometry detected that apoptosis levels were increased in siHOTAIR group. The level of cell senescence was higher in the siHOTAIR group than that of control group. Results of IHC indicated that Ki-67 and Bcl-2 protein of tumor tissue were inhibited, while BAX and cleaved caspase- 3 protein expressions were elevated simultaneously in the siHOTAIR group. TUNEL assay suggested that more apoptosis was observed in siHOTAIR group. Conclusion HOTAIR can affect proliferation and apoptosis of tongue squamous cancer cells. HOTAIR may be one of the new candidate targets for human tongue cancer therapy. Related Articles | Metrics

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Study on p21Gene Knock Out in G401 Cell Line by Using CRISPR/Cas9 System ZHAO Xiujuan, CHEN Wanbiao, ZHANG Peitao, ZHANG Na, CHU Xiaowen, BAI Xiangyang, YANG Bing, WU Xudong, WANG Xi 2016, 44 (10):  1190-1194.  doi: 10.11958/20160731 Abstract ( 926 )   PDF (548KB) ( 4416 )   Objective To knock out p21 gene in human malignant rhabdoid tumor（MRT） cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene， which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group， the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors ． Related Articles | Metrics

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The preparation of the mouse monoclonal antibodies specific for the DNAH2 protein CHANG Lixian, ZHANG Jingliao, REN Yuanyuan, SUN Congcong, WAN Yang, AN Wenbin, 2016, 44 (10):  1195-1198.  doi: 10.11958/20160317 Abstract ( 988 )   PDF (507KB) ( 3874 )   Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein， axonemal， heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1- 300 aa), in E. coli was constructed. IPTG was used to induce the expression of Hisimmunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal antiDNAH2 antibody is prepared successfully. Related Articles | Metrics

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Effects of D-tryptophan on biofilm formation and dispersal in Streptococcus mutans YANG Xiaoyue, LIAO Xiaohui, YE Jing, SHAO Can, WANG Bin, LIU Ying 2016, 44 (10):  1199-1203.  doi: 10.11958/20160341 Abstract ( 1072 )   PDF (692KB) ( 3791 )   Objective To investigate the effects of D-tryptophan (D-Trp) on the formation of Streptococcus mutans (S. mutans) biofilm and the dispersal of 24 h-old biofilm, and the drug susceptibility of S. mutans against chlorhexidine (CHX) under the role of D-Trp. Methods Optical density assay was used to evaluate the growth curve of S. mutans exposed to 5.0 mmol/L D-Trp for 28 h. The non-treated group was not added with D-Trp. After treatment with 1.0, 2.5 and 5.0 mmol/L D-Trp, crystal violet staining was used to observe the changes of S. mutans biofilm formation in treatment group and nontreatment group. Crystal violet staining and confocal laser scanning microscopy (CLSM) were applied to illustrate the effects of 1.0, 2.5 and 5.0 mmol/L D-Trp on the dispersal of 24 h-old S. mutans biofilm. Resazurin sodium was used to indicate the effect of 5.0 mmol/L D- Trp on the minimum inhibitory concentration (MIC) and the minimum biofilm inhibitory concentration (MBIC) of treatment groups and negative control group. Results The growth curves of planktonic S. mutans within 28 h was consistent in treatment group and the non-treated group, both attained exponential phase after 4 h and reached stationary phase at 22 h. Notably, when compared with non-treated group, the biomass of S. mutans biofilm was increased with time from 0 to 72 h after treatment with 1.0, 2.5 and 5.0 mmol/L D-Trp. And at the same time point, the biomass was significantly less in each subgroup of treatment group than that of non-treated group (P＜ 0.05). Crystal violet staining demonstrated that values of biomass（OD570） were less in treatment groups treated with 1.0, 2.5 and 5.0 mmol/L DTrp than those of non-treated group (P＜ 0.01). CLSM also showed that bacteria was adhered to the surface of media in treatment groups treated with 1.0, 2.5 and 5.0 mmol/L D-Trp. The values of biomass were lower in treatment groups than those of non-treated group (P<0.01). The MIC against S. mutans was 0.073 mg/L in both experimental group and negative control group. The values of MBIC were 0.293 mg/L and 2.344 mg/L in experimental group and negative control group, respectively. Under the action of 5.0 mmol/L D-Trp, the MBIC of S. mutans was reduced to 1/8. Conclusion Results indicate that D-Trp may inhibit the formation of S. mutans biofilm and promote the dispersal of biofilm already formed. DTrp may further help CHX exert its bactericidal activity to S. mutans. Related Articles | Metrics

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Detection of virulence gene and molecular typing of Shigella boydii isolated from clinical sources SUN Yang, ZHANG Haojie, GUO Wenxue, WANG Zhe, JIA Yuchi, QI Wei 2016, 44 (10):  1204-1208.  doi: 10.11958/20160440 Abstract ( 1018 )   PDF (403KB) ( 3851 )   Objective To understand genetic distribution, drug resistance, molecular typing and the epidemiological relativeness between strains of the Shigella boydii virulence. Methods Nine Shigella boydii strains were isolated form stool samples of patients with diarrhea from the Enteric Disease Clinic of the Second Hospital of Tianjin Medical University in June - October 2015. The strains were identified by biochemical test and serum agglutination test. Antibiotics susceptibility test was carried out using the Kirby- Bauer method. Polymerase chain reaction was used for detecting virulence genes. Pulsed- field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) technique were used to determine the epidemiological relationship between nine Shigella boydii strains. Results There were three subtypes in nine isolated Shigella boydii samples, including one, three and five isolates in Ⅰ , Ⅱ , Ⅳ subtypes respectively. All of the 9 isolates were multi-drug resistant. The resistant rate of these strains for ampicillin was 100% (9/9), and then the resistant rates of these strains for ceftazidime, streptomycin, gentamicin, trimethoprim/sulfamethoxazole, cefotaxime, ceftriaxone, norfloxacin and levofloxacin were 1/9, 4/9, 4/9, 4/9, 5/9, 5/9, 6/9, 6/9 and 6/9, respectively. All of these strains were sensitive to amikacin, cefperazone-sulbactam and imipenem. The ipaH was carried by all the testing strains, and none of the strains carried the sen, set1A, set1B, ial, virA, icsA and SigA. The detective rates of pic, sepA and sat were 4/9, 5/9 and 7/9 strains, respectively. Nine shigella boydii strains were divided into 8 PFGE types. The similarity between the spectrums of PFGE was 63.21% - 100%. Multilocus sequence typing showed that six isolates were belonged to ST648, two isolates were ST131 and one isolate was ST10. Conclusion Nine isolates of Shigella boydii (divided into three subtyping) isolated from our hospital are multi-drug resistant and they have distant relationships, belonging to the dissemination of case. Related Articles | Metrics

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Expression and identification of VEGF165 in bone marrow mesenchymal stem cells of rhesus GUO Zaiyu, ZHANG Heliang, SHUI Tao, ZHANG Guozhe, ZHAO Weihua, CHEN Qian, HOU Yanwei 2016, 44 (10):  1209-1212.  doi: 10.11958/20160444 Abstract ( 573 )   PDF (487KB) ( 3789 )   Objective To detect the transferred vascular endothelial growth factor (VEGF) 165 gene expression in rhesus autologous bone marrow mesenchymal stem cells (MSCs), and to explore the functional viability of transgenic MSCs. Methods MSCs from rhesus bone were isolated by Ficoll, which were used to detect the phenotype. After the culturing, the expression vector pcDNA- eGFP- VEGF165 was transfected into bone marrow MSCs. Fluorescence microscope and flow cytometry were used to detect the enhanced green fluorescent protein (eGFP) expression. At the same time, the phenotype in transfected MSCs was also indentified. The VEGF165 expression level was detected by RT- PCR. Results The highly purified MSCs were collected successfully. The transfected MSCs and daughter cells showed expressions of eGFP and VEGF165, which also remained the characteristics of MSCs. Conclusion The VEGF165 gene that is transfected into MSCs can maintain characteristics of MSCs, and stably express foreign genes. Related Articles | Metrics

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Study on expression profile of mRNA in brain of pronuclear transfer mice LI Tianjie, CAO Yanxiang, JIN Xiaohu, ZHAO Hongcui, YU Yang, QIAO Jie 2016, 44 (10):  1213-1216.  doi: 10.11958/20160746 Abstract ( 829 )   PDF (717KB) ( 4178 )   Objective To investigate the expression profile of mRNAs in brain samples collected from pronuclear transfer (PNT) mice. Methods Female CD-1 mice were superovulated, and zygotes were collected after mating with adult male mice. Zygotes with two pronuclei were selected for pronuclear transfer manipulation, and then the reconstructed zygotes were transferred into the oviduct of pseudopregnant female mice. The infant mice obtained from pronuclear transfer were called PNT group, while the embryoes that were not performed pronuclear transfer was regarded as control group. Total RNA were extracted from brain samples of both PNT and control mice, and cDNA were labeled with fluorescent dye. Genes that were differentially expressed were identified using the Agilent mouse mRNA array. Gene ontology analysis and pathway analysis were also completed. Results Compared with control group, 392 mRNAs were expressed differentially, which showed more than 2.0 times variation and statistical significance, accounting for 1.7% of all mRNAs. Among those 366 mRNAs were up-regulated and 26 mRNAs were down-regulated. Eleven mRNAs came to 4.0 times variation in total. Gene ontology analysis indicated that differentially expressed genes were significantly enriched in alternative mRNA splicing, small GTPase mediated signal transduction, regulation of insulin receptor signaling pathway, hydrolase activity, transmembrane transporter activity and pyrophosphatase activity. Significant enriched pathway terms contained ion channel transport, fatty acid metabolism, butanoate metabolism, triacylglycerol and ketone body metabolism. Conclusion Pronuclear transfer might influence some key metabolism process in mouse brain. Related Articles | Metrics

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The role of miR-34a in AngⅡ induced myocardial hypertrophy via alteration of AMPK/mTOR signal pathway HUANG Jionghua, LIN Yuhui, DAI Wenjun, WU Jinlei, XIE Wenjie, CHEN Yongquan 2016, 44 (10):  1217-1220.  doi: 10.11958/20160435 Abstract ( 1112 )   PDF (609KB) ( 3730 )   Objective To explore the effects of miR- 34a and the alteration of AMPK/mTOR signal pathway on angiotensin (Ang)Ⅱ -stimulated cardiomyocytes hypertrophy. Methods Neonatal rat cardiomyocytes were cultrued in vitro and were divided into 3 groups: negative control for lentivirus carrying miR-34a group (NC), AngⅡ plus lentivirus carrying negative control group (AngⅡ +NC) and AngⅡ plus lentivirus carrying miR-34a group (AngⅡ +miR-34a). The relative cell area was detected by confocal microscopy. The expression of miR-34a and hypertrophy-related genes (ANP and β-MHC) were analyzed by real time PCR. The AMPK/mTOR signal pathway was measured by Western blot assay. Results Compared to NC group, the relative cell area was increased in AngⅡ +NC group (P＜ 0.05). But compared with AngⅡ +NC group, the relative cell area was decreased in AngII+miR- 34a group (P＜ 0.05). Moreover, compared with NC group, the expression of miR- 34a was decreased, and the expression of hyperthophy- related genes(ANP and β- MHC) was upregulated in AngⅡ +NC group. However, the expression of miR- 34a was decreased, and the expression of hyperthophyrelated genes (ANP and β-MHC) was down-regulated (P＜ 0.05). Finally, compared to NC group, the ratio of phosphopAMPK/AMPK was significantly induced in AngII + NC group, but the ratio of phosphop-mTOR/mTOR was significantly decreased (P＜ 0.05). However, compared to Ang Ⅱ + NC group, the ratio of phosphop- AMPK/AMPK was significantly decreased in AngII + miR- 34a group, but the ratio of phosphop- mTOR/mTOR was significantly increased (P＜ 0.05). Conclusion miR-34a is able to inhibit myocardial hypertrophy of neonatal rat cardiomyocytes, and its mechanism is partly carried out by the alteration of the signal pathway of AMPK/mTOR. Related Articles | Metrics

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Geraniol attenuates vascular smooth muscle cell proliferation in rat carotid artery balloon injury model JIANG Jinyao, XU Xizi 2016, 44 (10):  1221-1224.  doi: 10.11958/20160290 Abstract ( 1139 )   PDF (571KB) ( 3685 )   Objective To evaluate the effects of geraniol(GOH) on neointima hyperplasia in rat carotid artery balloon injury model, and explore the potential molecular mechanisms associated with this effect. Methods Totally 20 male Sprague-Dawley (SD) rats were randomly divided into sham operation group (without balloon injury), control group (with balloon injury), low concentration group (with 50 mg/kg GOH intervention after balloon injury) and high concentration group (with 200 mg/kg GOH intervention after balloon injury). The intima to media (I/M) area ratio of neointima was measured by hematoxylin- eosin (HE) staining. The expression of proliferating cell nuclear antigen (PCNA) was measured by immunohistochemical staining at 14th day after operation. As the marker of oxidative stress, the levels of 8-hydroxy- 2’ - deoxyguanosine (8-OHdG) and malondialdehyde (MDA) were measured by enzyme linked inmmnosorbent assay (ELISA). Results The I/M ratio, IOD, 8-OHdG and MDA values were increased in control group compared with those of sham group. The I/M ratio, IOD and 8-OHdG values were reduced in low concentration group compared with those of control group. But there was no significant difference in MDA level between low concentration group and control group. The I/M ratio, IOD, 8- OHdG and MDA values were significantly reduced in high concentration group compared with those of control group, which showed a more significant inhibitory effect than that of low concentration group (P＜ 0.05). Conclusion GOH could attenuate balloon iniury induced neointima hyperplasia, which might be related to its effect on inhibiting expression of PCNA and decreasing oxidative stress. Related Articles | Metrics

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Activity study of the ethanol extract of thunberg fritillary leaf and the ethanol extract of thunberg fritillary flower on relieving cough, abolishing phlegm and relieving asthma WANG Hanhua, YANG Xiaochun, CUI Mingchao 2016, 44 (10):  1225-1228.  doi: 10.11958/20160345 Abstract ( 1615 )   PDF (382KB) ( 3952 )   Objective To observe the effects of the ethanol extract of thunberg fritillary leaf (EETFL) and the ethanol extract of thunberg fritillary flower (EETFF) on relieving cough, sputum elimination and relieving asthma. Methods The cough relieving effects of EETFL and EETFF were studied in mouse cough model caused by ammonia water and in guinea pig cough model caused by citric acid. The sputum elimination effects of EETFL and EETFF were researched by the observation of tracheal phenol red shedding in mice. The asthma relieving effects were tested by spraying method in guinea pigs. Results EETFL can obviously inhibit the incubation period and cough frequency of the model mice and guinea pigs induced by ammonia water and citric acid (P<0.05), and significantly improve the tracheal phenol red excretion volume in mice (P<0.05), and obviously prolong the incubation period of asthma (P<0.05). EETFF can obviously inhibit the incubation period and cough frequency of the model mice and guinea pigs induced by ammonia water and citric acid (P<0.05), and significantly improve the tracheal phenol red excretion volume in mice (P<0.05), but EETFF couldn’ t prolong the incubation period of asthma evidently. Conclusion EETFL has obvious activity of relieving cough, eliminating phlegm and relieving asthma. EETFF has obvious activity of relieving cough and eliminating phlegm, but EETFF has no anti-asthmatic activity under the current dose. Related Articles | Metrics

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Study of the feasibility on medical gas suction feeding device for replicating model of respiratory failure in dogs WANG Herong, WU Qi, SUN Xin, FU Shasha 2016, 44 (10):  1229-1232.  doi: 10.11958/20160029 Abstract ( 778 )   PDF (378KB) ( 3704 )   Objective To investigate the feasibility and stability of medical gas suction feeding device for copying the model of respiratory failure in dogs. Methods The self-designed medical gas suction feeding device was used to establish the model of respiratory failure in dogs. After regulating the proportion of oxygen (O 2), carbon dioxide (CO2) and nitrogen (N2), the fraction of inspiration O2 (FiO2) was decreased, while the forced inspiratory carbon dioxide (FiCO2) was increased. At the same time, the breathing out pathway was open to make the arterial oxygen partial pressure p (O2) and arterial blood carbon dioxide partial pressure p (CO2) reached and stabilized in a stable condition. The arterial blood samples between default state and 1 hour, 2 hour after modeling were collected to detect the blood gas index. Results One hour after the establishment of model, the p (O2) was ≤ 60 mmHg (1 mmHg=0.133 kPa), and p (CO2) was ≥50 mmHg (1 mmHg=0.133 kPa), which suggested that the dog model of respiratory failure was established successfully. Compared with the basic state, data of pH, oxygen content, oxygen saturation (SaO 2) and residual alkali (BE) were decreased at 1 hour and 2 hour after modeling (P < 0.05). The p (O2), oxygen content, SaO2 were decreased in 2 hour after modeling compared with those of 1h after modeling (P < 0.05). There were no significant differences in pH, p (CO2), TCO2, HCO3- and BE between 1 hour and 2 hour after modeling (P > 0.05). Conclusion Under the regulation of the medical gas suction feeding device, the respiratory failure state of large experimental animals is successfully established. Related Articles | Metrics

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The pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats JIN Tao, HU Yu, LIU Chao 2016, 44 (10):  1233-1237.  doi: 10.11958/20160501 Abstract ( 843 )   PDF (332KB) ( 3789 )   Objective To explore the pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats. Methods Ninety SD rats were randomly divided into three experimental groups: sham operation group （group A）, warm hepatic ischemia/reperfusion group（group B and group C）. Group C was given ischemic preconditioning treatment. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected 0 h, 2 h, 6 h, 12 h and 24 h after ischemia reperfusion injury. Levels of TNF- α and IL- 1β were tested detected by ELISA. Levels of malondialdehyde (MDA) and superoxide dismutase (SOD) of hepatocytes were detected at the same time points. Mitochondrial membrane potential was examined to assess ischemia reperfusion injury of hepatocytes in rats using chart of intensity of JC-1 in mitochondria. Results The serum levels of ALT, TNF-α, IL-1β, and MDA were significantly higher in hepatic warm ischemia reperfusion group and ischemic preconditioning group than those in sham operation group (P＜ 0.05). Values of prothrombin activity and cholinesterase were significantly lower in liver warm ischemia reperfusion group and ischemic preconditioning group than those of sham operation group (P＜ 0.05). The SOD level of liver was significantly lower in warm ischemia reperfusion group and ischemic preconditioning group than that in sham operation group. The indexes were better in ischemic preconditioning group than those of warm ischemia reperfusion group (P＜ 0.05). The mitochondrial membrane potential level of liver cells reached the lowest value 0 hours after ischemia and reperfusion, and then increased gradually within 24 hours (P＜ 0.05). And the level of mitochondrial membrane potential of liver cells was significantly higher in ischemic preconditioning group than that in warm ischemia reperfusion group (P＜ 0.05). Conclusion Ischemic preconditioning may play a protective role in warm ischemia- reperfusion injury in rats. Ischemic preconditioning may significantly decrease the levels of ALT, AST, TNF-α, IL-1β and MDA, and increase the SOD activity in hepatocytes. The damage of mitochondrial membrane potential is decreased after ischemic preconditioning, which might be the pathogenesis of ischemic preconditioning to warm ischemia reperfusion injury of hepatocytes in rats. Related Articles | Metrics

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Protective effects of P2X7R antagonist on chronic pancreatitis in mice ZHANG Guixian, LIU Dawei, NIE Wei, ZHANG Yi, LIU Weiyun, LIU Hongbin 2016, 44 (10):  1238-1242.  doi: 10.11958/20160679 Abstract ( 987 )   PDF (2020KB) ( 3906 )   Objective To investigate the role of purinergic 2X7 receptor (P2X7R) and its downstream target-NLRP3 inflammasome activation in the process of pancreatic fibrosis in a mouse model of chronic pancreatitis (CP). Methods Forty C57BL/6 mice were randomly divided into normal control group, CP group, P2X7R antagonist oxidized ATP (OxATP) group and brilliant blue G (BBG) group. The chronic pancreatitis model was induced by repeated intraperitoneal injection of the cholecystokinin analogue caerulein with the dose of 50 μg/kg for six weeks. Normal saline, OxATP (20 μL, 300 μmol/L) or BBG (20 μL, 10 μmol/L) were administered for CP group, OxATP group and BBG group for two weeks after the last caerulein injection. Then all mice were sacrificed and the histopathological changes of the pancreas, especially the fibrotic degrees were evaluated by HE stain, fibrosis score, Sirius red staining and α-SMA immunohistochemical stain. The pancreatic P2X7R, NLRP3 and Caspase-1 expressions were detected by immunohistochemistry respectively to compare the changes between the groups, and explore the role of P2X7R-NLRP3 signaling pathway in pancreatic fibrosis. Results Compared with the normal control group, the scores of pancreatic fibrosis and the expressions of P2X7R, NLRP3 and Caspase- 1 in pancreas were significantly increased in CP model group (P＜ 0.05). Compare to CP group, the pancreatic chronic inflammation and the fibrosis indices such as HE fibrosis score, Sirius red staining and α- SMA immunohistochemical stain were ameliorated obviously in OxATP and BBG groups (P＜ 0.05). And expressions of P2X7R, NLRP3 and Caspase-1 in the pancreas were all reduced greatly in both OxATP and BBG groups (P<0.05). Conclusion P2X7R antagonist OxATP and BBG can significantly decrease pancreatic chronic inflammation and fibrosis in the mouse model of CP, which suggests that the blockade of P2X7R NLRP3 inflammasome signaling pathway may represent a novel therapeutic strategy for CP and its fibrotic process. Related Articles | Metrics

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The relationship of the folate metabolism related gene polymorphisms of MTHFR and MTRR with unexplained recurrent spontaneous abortion XIE Xiaoyuan, ZHANG Ying, XIN Li, LENG Junhong, LU Yanqiang, XUE Yan, ZHU Xiulan 2016, 44 (10):  1243-1246.  doi: 10.11958/20160099 Abstract ( 1429 )   PDF (343KB) ( 4675 )   Objectives To investigate the relationship of methylenetetrahydrofolate reductase (MTHFR) and methioninesynthase reductase (MTRR) with unexplained recurrent spontaneous abortion (URSA). Methods Case control study was used to select 244 patients with URSA (miscarriage group) and 116 normal women (control group) who were admitted to Tianjin Medical University General Hospital and Tianjin Women’ s and Children’ s Health Center from January 2013 to March 2015. The oral mucosal epithelial cells were extracted using fluorescence quantitative PCR to detect MTHFR gene C677T, A1298C and MTRR gene loci of A66G single nucleotide polymorphisms (SNP). The relationship between folate metabolism related gene polymorphisms of MTHFR and MTRR and URSA was analysed. Results The frequency of C677T genotype MTHFR was significantly higher in URSA group than that in the control group, and the frequency of CT genotype was significantly lower than that of the control group (P<0.05). There was no significant difference in the frequencies of A1298C MTRR and A66G MTHFR between the two groups. The activity of MTHFR, red cell folate and plasma folate levels were significantly lower in URSA group than those of control group. Homocysteine levels were significantly higher in URSA group than those of control group (P<0.05). There were no significant differences in serum folic acid, red cell folate, homocysteine cysteine levels between patients <35 years old and ≥ 35 years old in URSA group. Conclusion C677T MTHFR gene polymorphism is associated with unexplained recurrent spontaneous abortion . Related Articles | Metrics

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Analysis of the quality of life in patients with severe acute respiratory syndrome for 10 years HUA Jingna, SUN Xin, WU Qi， LI Li, XING Zhiheng, ZHAO Xinqian 2016, 44 (10):  1247-1250.  doi: 10.11958/20160361 Abstract ( 1212 )   PDF (336KB) ( 3693 )   Objective To analyse the quality of life in patients with severe acute respiratory syndrome (SARS) for 10 years. Methods Twenty-five SARS patients were collected as SARS group and 25 healthy persons were used as control group. There were no significant differences in general clinical data between two groups. The quality of life of the respondents was evaluated by the MOS item short form health survey (SF- 36) including physiological function, physical pain, general health condition, energy, social function, emotional function and mental health. The pulmonary function indicators including forced vital capacity (FVC), one second forced expiratory volume (FEV1), one second rate (FEV1/FVC), total lung capacity (TLC), residual volume (RV), corrected breath diffusing capacity (TLCOc SB) and corrected dispersion rate TLCOc/VA were also compared between two groups. The correlation between each dimension score of pulmonary function indexes and quality of life were analyzed in SARS group. Data of hip and femoral head necrosis were detected by MRI scan in SARS group of patients. Results Total quality of life score was poor in 23 cases (92.0%), and moderate quality of life score in 2 cases (8.0%). The quality of life score was lower in SARS group than that of control group (P < 0.01). Data of TLCOc SB was lower in SARS group than that of control group. The score of physiological function was positively correlated with FEV1 and FEV1/FVC. The score of physical pain was positively correlated with FEV1/FVC (P＜ 0.05). There were no significant differences between quality of life score and pulmonary function indicators. There were 11 cases (44% ) of avascular necrosis of femoral head, and 14 cases (56%) of non-avascular necrosis of femoral head in SARS group. The score of physiological function was lower in femoral head necrosis group than that of non necrosis group (P＜ 0.05). Conclusion The lower quality of life in patients with SARS is not only related with psychological factors but also related with physical factors. The damage of pulmonary function and (or) necrosis of femoral head affect the quality of life in patients with SARS, especially the physiological function. Related Articles | Metrics

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The change of AChE related microRNAs derived from peripheral blood mononuclear cells in acute ischemic stroke HAN Bin, MA Xiaofeng, ZHANG Chao 2016, 44 (10):  1251-1254.  doi: 10.11958/20160518 Abstract ( 946 )   PDF (329KB) ( 4121 )   Objective To investigate the expression changes of acetylcholinesterase (AChE) related microRNAs derived from peripheral blood mononuclear cells (PBMCs) in patients with stroke. Methods The microRNAs for targeting AChE mRNA were selected via prediction software and previous studies. PBMCs were extracted from venous blood samples of acute ischemic stroke patients (onset < 24 h) and healthy controls. The expressions of microRNAs and AChE mRNA were quantified using real-time PCR (RT-PCR). The protein level of AChE was detected by Western blot assay. Results The predicted microRNAs included microRNA (miR) - 24, -28, -124, -132, -182*, -194 and -484. The expression levels of miR-24, -124, -132 and-194 were significantly elevated in stroke patients compared with those of controls (P<0.05). There were no significant changes in expression levels of miR-28, -182* and -484. Additionally, the relative expression levels of intracellular AChE mRNA and protein decreased significantly in stroke patients (P<0.05). Conclusion MiRNAs can enhance cholinergic anti-inflammatory pathway by targeting AChE in patients with acute ischemic stroke. Related Articles | Metrics

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The clinical value of bedside testing of plasma levels of NT-proBNP and cTnI in the rapid diagnosis of cardiac dyspnea LIAO Tong, ZHANG Yuanchun, LI Huanlun, GU Zhentuo, PAN Chaoqing, LU Lihua, TAN Zhiwei, HUANG Xifan 2016, 44 (10):  1255-1258.  doi: 10.11958/20160114 Abstract ( 986 )   PDF (309KB) ( 3649 )   Objective To explore the diagnostic value of combined bedside detection of aminoterminal pro-B-type natriuretic peptide (NT-proBNP) and cardiac troponin I (cTnI) in the diagnosis of cardiac dyspnea. Methods A total of 120 patients with dyspnea admitted in our department from June 2014 to February 2016 were included in this study. At the time of admission, NT-proBNP and cTnI levels were measured by bedside test. Values of NT-proBNP＞ 300 ng/L or cTnI＞ 0.16 mg/L were defined as positive for cardiac dyspnea. According to the final diagnosis, patients were divided into two groups: cardiac dyspnea group (n=68) and pulmonary dyspnea group (n=52). At the same time, 30 healthy people were selected as control group. Values of NT-proBNP and cTnI were used for statistical analysis between the three groups. The sensitivity and specificity of NT-proBNP, cTnI and cTnI+NT-proBNP were compared between three groups. Results The levels of NTproBNP and cTnI were significantly higher in pulmonary dyspnea group and cardiac dyspnea group than those in the control group, and the levels were significantly higher in cardiac dyspnea group than those of pulmonary dyspnea group (P<0.01). The detection sensitivity of NT-proBNP and cTnI alone was 67.65% and 52.94%, combined detection of both was up to 94.12%. The specificity of NT-proBNP and cTnI detection alone was 70.00% and 53.33%, respectively, and combined detection of both was up to 86.67%. The sensitivity of NT-proBNP+cTnI was significantly higher than that of NT-proBNP and cTnI alone (P＜ 0.05), but there was no significant difference in the specificity between combined detection and individual detection of NT-proBNP. The positive predictive value of the combined detection in the diagnosis of cardiac dyspnea was 94.12% (64/68), and the negative predictive value was 86.67% (26/30). Conclusion Bedside detection with combination of cTnI and NT-proBNP has important clinical application value in the rapid diagnosis of cardiac dyspnea, which is a rapid clinical testing method Related Articles | Metrics

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In vitro activity of amikacin combined with the ingredient of coptis chinensis against multiple drug resistance of Escherichia coli#br# #br# SHUAI Lihua,JIANG Dengzhao, LIU Huai, ZHOU Xiaojuan, PAN Bin, YE Xingwen 2016, 44 (10):  1259-1262.  doi: 10.11958/20160547 Abstract ( 1214 )   PDF (337KB) ( 3697 )   Objective To observe the antibacterial effects of amikacin sulfate combined with the ingredient of coptis chinensis： the extract of coptis chinensis, berberine hydrochloride and palmatine hydrochloride on multiple drug resistance (MDR) of Escherichia coli (E.coli) in vitro. Methods One hundred strains of E.coli checked out from 2013 to 2014 that showed resistence to cefotaxime were selected, and in which MDR to bacteria were determined. The extended-spectrum β- lactamases (ESBLs)-producing by Kirby-Bauer test. The minimum inhibitory concentrations (MIC) of the extract of coptis chinensis, berberine hydrochloride, palmatine hydrochloride and amikacin sulfate on ESBLs- producing E.coli were determined firstly, and then the sterilization effects of amikacin sulfate combined with the other three medicines were observed by broth microdilution checkerboard method together with their fractional inhibitory concentrations (FIC), with ATCC 25922 for quality control strains. Results Ten MDR E.coli were screened and proved to be ESBLs-producing. The inhibitory effects were enhanced in a synergistic or additive pattern when amikacin sulfate combined with the extract of coptis chinensis, berberine hydrochloride on nine of the ten MDR E.coli in vitro, where the inhibitory effects were a synergistic or additive pattern when amikacin sulfate combined with palmatine hydrochloride on all of the MDR E.coli in vitro. Conclusion Palmatine hydrochloride or berberine hydrochloride or the extract of coptis chinensis combined with the amikacin sulfate has significant value in treatment of MDR E.coli, which is worthy of further study. Related Articles | Metrics

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Study on relationship between plasma homocysteine, Kv1 .3 channel and troponin in patients with acute ST-segment elevation myocardial infarction WANG Yuling, FENG Jianyu, ZENG Xiangfei, YANG Shaobing, YAN Ning, JIA Shaobin 2016, 44 (10):  1263-1267.  doi: 10.11958/20160128 Abstract ( 1084 )   PDF (354KB) ( 3728 )   Objective To investigate the relationship between plasma homocysteine (Hcy), Kv1.3 channel and cardiac troponinI (cTnI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods According to the level of Hcy, 80 STEMI patients were divided into STEMI with Hhcy group (Hcy > 15 μmol/L, n=41) and control group (STEMI group, Hcy≤15 μmol/L, n=39). The Hcy, blood lipid and cTnI were detected with automatic biochemistry analyzer, respectively. Peripheral lymphocytes were isolated by ficoll density gradient centrifugation. Real-time PCR was used to detect mRNA expression of Kv1.3, and Western blot assay was used to detect protein expression of Kv1.3. Results cTnI concentrations were obviously higher in STEMI with Hhcy group than those in STEMI group（μg/L： 22.997 ± 5.880 vs. 12.881 ± 6.343； P＜ 0.01）. Multiple linear regression analysis showed that age, gender, hypertension, diabetes mellitus, smoking, family history, total cholesterol (TC), triglyceride (TG), high density lipoprotein- cholesterol (HDL- C) and low density lipoprotein-cholesterol (LDL-C) had no obvious influence on Hcy (P＞ 0.05). The relative expression levels of Kv1.3 mRNA and protein were significantly higher in STEMI with Hhcy group (1.35±0.14, 0.85±0.12) than those in STEMI group (1.00± 0.07, 0.64± 0.05, P＜ 0.05). Moreover, there was a positive relation between Hcy level and the mRNA and protein expression of Kv1.3 channel (r=0.299, r=0.542, P＜ 0.05). There was a positive relation between protein expression levels of Kv1.3 channel and cTnI (r=0.644, P＜ 0.05). Conclusion Our results support that Hcy could exacerbate the concentration of cTnI through playing an important role in the Kv1.3 mRNA and protein expression in lymphocytes. Related Articles | Metrics

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The influence and clinical significance of Single and Dual-chamber Pacing on Central Aortic Pressure and Augmentation Index in non-smoking individuals MIAO Shuai, LI Guangping, YE Lan, YAN Zhehui 2016, 44 (10):  1268-1271.  doi: 10.11958/20160530 Abstract ( 1047 )   PDF (303KB) ( 3897 )   Objective To investigate the influence and clinical significance of single and dual- chamber pacing on central aortic pressure (CAP) and augmentation index (AI) in non-smoking individuals. Methods Totally, 83 non-smokers with pacemaker-implanted were consecutively enrolled in this study, and they were divided into three groups: dual-chamber pacemaker group (DDD, n=35), single-chamber pacemaker group (VVI, n=33) and control group (n=15). Heart rate (HR), CAP, AI, systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured in three groups of patients. Finally, DDD pacing mode was turned into VVI pacing mode in patients of DDD group and the indexes were measured again. All of the indexes were recorded and analyzed. Results There were no significant changes in baseline characteristics and laboratory data between three groups (P＞ 0.05). Left atrial diameters were significantly higher in VVI group than those of control group (P＜ 0.05). Values of CAP were significantly higher in DDD group than those of control group and VVI group (P＜ 0.05). Values of AI, corrected AI (AIc) and brachial BP were significantly higher in DDD group than those of VVI group (P＜ 0.05). Values of CAP and brachial BP were significantly lower in VVI group than those of control group (P＜ 0.05), while no significant changes were found in AI and AIC between VVI group and control group (P＞ 0.05). All of these indexes (CAP, AI and brachial BP) were significantly reduced after the pacing mode was changed (P＜ 0.05). Conclusion In nonsmokers, dual-chamber pacing mode can increase CAP and AI. Related Articles | Metrics

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The clinical efficacy of thrombus aspiration in ST-elevation myocardial infarction patients undergoing primary percutaneous coronary intervention LI Zhenlong, WANG Yuping, ZHANG Tong, WANG Yuanyuan, MA Hongjun, LIU Yin 2016, 44 (10):  1272-1275.  doi: 10.11958/20160343 Abstract ( 848 )   PDF (308KB) ( 3813 )   Objective To explore the clinical efficacy and outcomes of thrombus aspiration in ST-elevation myocardial infarction (STEMI) patients undergoing percutaneous coronary intervention (PPCI). Methods A total of 664 cases of STEMI patients undergoing PPCI from Tianjin Chest Hospital from January 2013 to March 2015 were retrospectively analyzed. Patients were divided into two groups: primary PCI after thrombus aspiration group and conventional primary PCI without thromobus aspiration group. The base- line clinical characteristics, and the procedure of percutaneous coronary intervention were similar between two groups. The TIMI flow, LVEF, one-month outcomes and 12-month outcomes were compared between two groups. Results There were significant higher TIMI flow, LVEF in primary PCI after thrombus aspiration group than those of conventional primary PCI without thromobus aspiration group (P＜ 0.05). At one-month followup, there were no significant differences in major adverse cardiac events (MACE) between the two groups. At 12-month follow- up, there were no significant differences in cardiovascular death, recurrent myocardial infarction and new hospitalization between two groups. The incidence of recurrent angina was significantly higher in non-aspiration group than that in aspiration group (P＜ 0.05). There were the better survival functions without MACE in aspiration group than that of control group. Conclusion Thrombus aspiration, as an adjunctive method to primary PCI for STEMI, may improve TIMI flow, have beneficial effects on LVEF and reduce the incidence of recurrent angina at 12-month follow up. Related Articles | Metrics

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Feasibility and safety of McKeown-type minimally invasive esophagectomy in para-recurrent laryngeal nerve lymphadenectomy XUE Yabin, LYU Hongbo, DU Tongxin, SUN Wei 2016, 44 (10):  1276-1279.  doi: 10.11958/20160351 Abstract ( 1227 )   PDF (292KB) ( 3758 )   Objective To explore the feasibility and safety of Mckeown-type minimally invasive esophagectomy in para-recurrent laryngeal nerve lymphadenectomy. Methods A total of 163 cases underwent minimally invasive McKeown resection for esophageal carcinoma in the Affiliated Tumor Hospital of Xinjiang Medical University were retrospectively analyzed. Patients were divided into routine treatment group (n=63), right recurrent nerve lymph node dissection group (right group, n=53) and bilateral recurrent laryngeal nerve lymph node dissection group (bilateral group, n=47) according to the operation modes. The postoperative pathology, operation time, intraoperative bleeding volume, postoperative hospitalization time, number of lymph nodes and pneumonia, anastomotic fistula, laryngeal nerve injury and other complications were compared between three groups of patients. Pathological conditions were consisted of the location of tumor, the degree of differentiation, T stage and pathological stage. Postoperative complications were followed up for 6 months. Results There were no significant differences in tumor location, pathological differentiation degree, T stage and pathologic stage between three groups. The amount of bleeding was more in the routine group than that of the right group and the bilateral group (P＜ 0.05). There was no significant difference in the amount of bleeding between the right group and the bilateral group. The total lymph nodes and thoracic lymph nodes were increased in order in routine group, the right group and the bilateral group, and there was significant difference between three groups(P＜ 0.05). There were no significant differences in other operation data and complications between three groups. Conclusion McKeown- type minimally invasive esophagectomy shows good feasibility and safety for para-recurrent laryngeal nerve lymphadenectomy . Related Articles | Metrics

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Study of different modes of delivery and outcome of pregnant women PANG Yunduo, MA Yan,DONG Shengwen, SUN Zhong 2016, 44 (10):  1280-1282.  doi: 10.11958/20160033 Abstract ( 978 )   PDF (264KB) ( 4206 )   Objective To study the different selections of delivery mode of pregnant women, and compare their outcomes. Methods Data of 7 509 pregnant women hospitalized in the General Hospital of Tianjin Medical University between January 2013 and December 2014 were retrospectively analyzed. Of them, there were 6 229 cases of primipara and 1 280 cases of multipara. The numbers of vaginal delivery and cesarean delivery were statistical analyzed between primipara and multipara. The cesarean delivery indications of multipara was also studied. The vaginal delivery rates and cesarean delivery rates, and their outcomes were compared between primipara and scar uterus maternal pregnancy. Results The cesarean delivery rate was higher in multipara than that of primipara (69.06% vs. 57.54%, χ2=59.19, P＜ 0.05). Among them, 631 cases were uterine scar pregnancy, 39 cases (6.18%) were vaginal delivery and 592 cases (93.82%) were cesarean delivery. The uterine scar was the main indicator for cesarean delivery in multipara. Compared the outcomes of cesarean delivery, the weight of neonatal body mass was relatively light, the hospital stay was shorter, the amount of 24 h postpartum bleeding and postpartum fever were less in vaginal delivery in primipara and pregnant women with uterine scar (P＜ 0.05). There were no significant differences in scores of newborn. There were no significant differences in outcomes of vaginal delivery between primipara and pregnant women with uterine scar. Compared outcomes of cesarean delivery between multipara with scar uterus and primipara, there were increased maternal blood loss and postpartum fever in multipara with scar uterus (P<0.05). Conclusion The cesarean delivery is still the mainly mode for multipara. The vaginal delivery is the best mode for pregnant women with scar uterus compared with that of primipara. The vaginal delivery is the safer way of delivery for pregnant women. Related Articles | Metrics

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Surgical treatment of adult mandibular condylar fractures: a clinical analysis of 32 cases YAN Yingbin, MU Jie, ZHANG Lancheng, LIU Hao, WANG Zhixing, ZHANG Jun, SHEN Jun, ZHANG Ping 2016, 44 (10):  1283-1286.  doi: 10.11958/20160606 Abstract ( 1164 )   PDF (1264KB) ( 4035 )   Objective To investigate the effect of surgical treatment of adult mandibular condylar fractures (including intracapsular fracture). Methods Thirty-two patients (33 sides) with condylar fractures underwent open reduction and rigid fixation. Six patients with intracapsular condylar fractures were treated with two 18- mm positional screws through a preauricular approach. Six patients (7 sides) with condylar neck fractures were rigidly fixed by 1 mini- plate via a retromandibular transparotid approach. Twenty patients with subcondylar fractures were operated and fixed by two titanium plates using an angular approach. Results The mean follow-up period was 13.5 months, and the mean maximum mouth opening was 37.5 mm by the last visit. All patients acquired good occlusal relationship and mandibular symmetry. Seven patients (21.9%) experienced transient palsy of the branches of the facial nerve, and recovered completely after three months. One patient developed a salivary fistula, and healed after two weeks of gauze compression. No permanent deficit of any facial nerve branch was observed. No patient showed condylar head resorption. Conclusion Appropriate surgical approaches and fixation methods for different types of condylar fractures are the key factors to achieve reliable clinical results. Related Articles | Metrics

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Research on the relationship between miRNAs and peritoneal fibrosis LI Dong， YAN Tiekun 2016, 44 (10):  1287-1290.  doi: 10.11958/20160541 Abstract ( 812 )   PDF (331KB) ( 3809 )   In recent years, microRNAs were found to be one of the key factors in post transcriptional gene regulation which are involved in occurrence,development of many diseases. Peritoneal dialysis has become an effective alternative treatment approaches for patients with ESRD. Peritoneal fibrosis is one of the most important factors forced patients with ESRD to interrupt from long-term peritoneal dialysis, hence restricts the application and development of peritoneal dialysis.MicroRNAs are closely related to peritoneal fibrosis. This article reviews the role of microRNAs in the pathogenesis and treatment of peritoneal fibrosis. Related Articles | Metrics

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One case with anti -S antibodies causing problems in cross-matching blood WEI Xiaojuan, WANG Wei, LIU Chenmei 2016, 44 (10):  1291-1292.  doi: 10.11958/20160305 Abstract ( 1027 )   PDF (256KB) ( 3744 )   Anti-S antibody is rare and irregular antibody in MNS blood group system. One patient was found positive when doing antibody screening experiment before coronary artery bypass grafting. This is the first case of serum IgG-anti-S in our laboratory. S-antibody screening test and irregular antibody identification are important before blood transfusion, which can reduce the transfusion reaction Related Articles | Metrics

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A case report of lipomatous hemangiopericytoma DONG Liru, REN Huiqiang, WANG Shan, SONG Xudong 2016, 44 (10):  1293-1294.  doi: 10.11958/20160339 Abstract ( 887 )   PDF (1104KB) ( 3809 )   Lipoma of hemangiopericytomas (LHPC) is one ofrare soft-tissue tumors that grow slowly and occur in deep soft tissues. The classical histological morphology of LHPC is circularor spindle undifferentiated tumor cells that grow surrounding the thin-walled branching blood vessels. The clinical and pathological features of one patient with LHPC are retrospective analyzed to strengthen the understanding of LHPC. Related Articles | Metrics

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A case report of anal melanoma LIU Jingzhou 2016, 44 (10):  1295-1296.  doi: 10.11958/20160415 Abstract ( 792 )   PDF (1085KB) ( 3763 )   Anal malignant melanoma is a rare disease with high malignancy and poor prognosis. This paper reviews the relevant literature of the diagnosis and treatment of a case of anal melanoma. Related Articles | Metrics