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15 July 2019, Volume 47 Issue 7 Previous Issue    Next Issue

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Construction of recombinant lentiviral vectors with rat paralemmin-3 gene and its expression in alveolar macrophages TANG Jian, CHEN Xu-xin, FAN Chong-yang, HAN Zhi-hai 2019, 47 (7):  673-677.  doi: 10.11958/20190650 Abstract ( 833 )   PDF (526KB) ( 5671 )   Objective To construct a recombinant lentiviral vector containing rat paralemmin-3 (PALM3) gene and detect its expression in alveolar macrophages (NR8383 cells). Methods Rat PALM3 gene fragments were amplified and obtained by polymerase chain reaction (PCR), and were cloned to pCV186-Cherry. After identification of pCV186-CherryPALM3 with PCR and DNA sequencing, the recombinant vector, pHelper 1.0 and pHelper 2.0 plasmids were co-transfected into 293T packaging cells to generate recombinant lentiviral vector lenti-CV186-Cherry-PALM3. The titer of lenti-CV186-Cherry-PALM3 was determined by fluorescence labeling method. NR8383 cells were transfected with different concentrations of lenti-CV186-Cherry-PALM3 to obtain the optimal MOI according to transfection efficiency. The protein expression of PALM3 was detected by Western blot assay following transfection with lenti-CV186-Cherry-PALM3 under the optimal MOI. Results Rat PALM3 gene fragments were obtained by PCR and ligated into the pCV186 plasmid vector.The PCR and DNA sequencing results demonstrated that the recombinant plasmid pCV186-Cherry-PALM3 was successfully constructed. The recombinant lentiviral vector was successfully packaged, and its titer was 3.0×108 TU/mL. The optimal MOI value for the NR8383 cells was 20. The result of Western blot assay showed that PALM3 protein expression was significantly enhanced after transfection with lenti-CV186-Cherry-PALM3 (P＜0.05). Conclusion The recombinant lentiviral vector lenti-CV186-Cherry-PALM3 has been constructed, and efficiently expressed in alveolar macrophages,which offers a basis for further research of the effect of PALM3 on macrophage polarization. Related Articles | Metrics

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Shh pathway mediated the potential of reactive astrocytes to acquire stem cell properties after injury XU Zi-, LI Xiao-hong, YE Yi-chao, LIU Xiao-yin, TU Yue 2019, 47 (7):  678-682.  doi: 10.11958/20181966 Abstract ( 729 )   PDF (684KB) ( 3755 )   Objective To explore the effect of sonic hedgehog (Shh) on the stem cell potential in cerebral cortical reactive astrocytes (RAS). Methods The astrocytes of cerebral cortex of SD rats were cultured in vitro. The inflammatory RAS were induced by tumor necrosis factor (TNF)-α, interleukin (IL)-1 α and C1q. The traumatic RAS were prepared by scratch test. The morphology and purity of cells were observed by immunofluorescence staining. The cells were divided into the normal astrocytes group (control group), the inflammation group, and the scratch group (trauma group). The inflammation group was cultured for 24 h after the intervention, control group and scratch group were cultured for 5 days. The positive rate of Nestin was observed by immunofluorescence staining. The expressions of Shh in different injured groups were detected by Western blot assay. Then, the cells in each group were replaced with neural stem cell culture medium on the 7th day, and cells were subdivided into the activation group (the group was replaced with neural stem cell culture medium and added with smoothened agonist) and inhibition group (the cells in each group were replaced with neural stem cell culture medium and then added with cyclopamine). After 14 days, the expression levels of RAS -induced neural stem cell related genes were verified by qRT-PCR. Results In cell injury model, the result of immunofluorescence staining showed that Nestin-positive cells were significantly more in the scratched group than those in the inflammatory group and control group. Western blot assay showed that the expression of Shh was significantly higher in the scratched group than that in the inflammatory group(P＜0.05). After replacing the neural stem cell conditioned medium, the expressions of neural stem cell genes (Nestin, Pax-6, Sox-2, Oct-4 and Map-2) were significantly higher in the scratch group than those in the inflammatory group and the control group (P＜0.01). After adding the cyclopamine, the gene expression levels were significantly reduced in the three groups, but the expressions of neural stem cell genes were significantly higher in the scratch group than those of the other two groups (P＜0.01). Conclusion In vitro, the Shh signaling pathway can mediate the potential of stem cells in the cerebral cortex RAS after injury. Related Articles | Metrics

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Construction of eukaryotic expression vector of HCV Core gene and its influence on autophagy of HeLa cells WANG Min-min, WEI Jian-hong, REN Lai-feng 2019, 47 (7):  683-687.  doi: 10.11958/20182209 Abstract ( 641 )   PDF (668KB) ( 3864 )   Objective To construct eukaryotic expression vector of hepatitis C virus (HCV) core protein, and investigate the effect of HCV Core protein on the autophagy of human cervical cancer HeLa cells. Methods The fragment of core was obtained by PCR from HCV replicon plasmid pJFH1. After purification and recovery, the DNA fragment was connected with the PLVX-IRES-ZsGreen1 vector using the homologous recombination method. DNA sequencing was used to verify the correctness of the sequence. PLVX-IRES-ZsGreen1-core was transfected into HeLa cells. Cells were divided into PLVX-IRES-ZsGreen1-core recombinant plasmid transfection group, PLVX-IRES-ZsGreen1 empty carrier transfection control group and control group without transfection plasmid blank. Western blot assay and immunofluorescence(IF) were applied to detect the expression of HCV Core protein and the autophagy-related biomarker (microtubuleassociated protein 1 light chain3, LC3) in HeLa cells. Results PLVX-IRES-ZsGreen1-core recombinant plasmid wasobtained. The sequencing results showed that the sequence of recombinant plasmid was completely corrected, and the HCV Core protein can be effectively expressed in HeLa cells. The results of Western blot assay and immunofluorescence also showed that LC3 Ⅱ protein level and LC3 foci were increased significantly in cells with HCV Core protein. The level of LC3 II was significantly higher in PLVX-IRES-ZsGreen1-Core recombinant plasmid transfection group (1.069±0.049) than that in PLVX-IRES-ZsGreen1 empty vector transfection control group (0.776 ± 0.047, P<0.05). Conclusion The eukaryotic expression vector of HCV Core is successfully constructed, and the HCV Core protein can induce autophagy in HeLa cells. Related Articles | Metrics

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The importance of cellular interaction in vascular endothelial injury induced by overlap syndrome and the role of intercellular adhesion molecule-1 XIAO FanCAO Jie, LI Xin 2019, 47 (7):  687-691.  doi: 10.11958/20190304 Abstract ( 626 )   PDF (384KB) ( 5102 )   Objective To explore the important role of neutrophil (PMN) and vascular endothelial cell (VEC) interaction in vascular endothelial injury induced by overlap syndrome and the relationship with the action of intercellular adhesion molecule (ICAM)-1 at the cellular level. Methods Samples of fasting peripheral venous blood were taken from 16 healthy volunteers, 2.5 mL per person, 40 mL were sampoed at each time and 10 times were sampoed successively. PMN was separated by double-layer density gradient centrifugation. Then the experimental group directly co-cultured with neutrophils and endothelial cells was placed in the cell culture incubator simultaneously with the control group of endothelial cells alone.The culture was carried out for 4 hours, and then the respiratory simulation system of the Department of Respiratory Medicine of Tianjin Medical University General Hospital was used for intermittent and continuous hypoxia exposure for 8 hours. After exposure, the supernatant and endothelial cells were collected separately. The inflammatory factors in the supernatant including interleukin (IL) - 6, tumor necrosis factor (TNF) - α, adhesion molecule ICAM-1, catalase (CAT)activity and malondialdehyde (MDA) concentration were detected. The expression levels of VEC pro-apoptotic gene Bak and anti-apoptotic gene Bcl-xl mRNA were detected by qRT-PCR. Results The concentrations of IL-6, TNF-α, ICAM-1 and MDA were significantly higher in the experimental group than those in the control group (P＜0.05). The activity of CAT was significantly lower in the experimental group than that in the control group (P＜0.05). The expression levels of Bak mRNA and Bcl-xl mRNA were significantly higher in the experimental group than those in the control group, while Bcl-xl/Bak was significantly lower than that in the control group (P＜0.05). In experimental group ICAM-1 was positively correlated with IL-6, TNF - α and MDA, but ICAM-1 was negatively correlated with CAT and Bcl-xl / BAK (P＜0.05). Conclusion The interaction between PMN and VEC aggravates vascular endothelial injury caused by overlap syndrome, and ICAM-1 plays an important role in it. Related Articles | Metrics

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The effect and mechanism of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells YUAN Le-yong, WANG Mei-fang 2019, 47 (7):  692-696.  doi: 10.11958/20182237 Abstract ( 639 )   PDF (617KB) ( 4374 )   Objective To investigate the effect of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells and its mechanism thereof. Methods Non-small cell lung cancer cell line SPC-A1 was cultured and interfered with OCCLUDIN expression by siRNA transfection. The cells were divided into blank control group (control group), idling group (siCtrl group)and siRNA transfection group (siOCLN group). The effects of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells were detected by CCK-8 assay, flow cytometry and Western blot assay. Results The cell proliferation abilities of siOCLN cells were significantly lower at 24 h, 48 h, 72 h, 96 h and 120 h than those of control group and siCtrl group (P＜0.05). The apoptosis rate was significantly higher in siOCLN group than that of control group and siCtrl group (P＜0.05). The levels of apoptosis-related proteins such as Bax, caspase-3, caspase-9, AIF and Cyt C were significantly higher in siOCLN group than those in control group and siCtrl group (P＜0.05). The level of anti-apoptotic protein Bcl-2 was significantly lower in siOCLN group than that of control group and siCtrl group (P＜0.05). At the same time, the phosphorylation levels of AKT and PI3K protein were significantly lower in siOCLN group than those in control group and siCtrl group (P＜0.05). Conclusion OCCLUDIN may promote the proliferation and inhibit apoptosis of SPC-A1 cells by regulating PI3K/AKT signaling pathway. Related Articles | Metrics

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Effects of β-PGG on glycolysis and proliferation of pancreatic cancer cells CHEN Xi-juan, HU Li-juan, QIU Shuai, YANG Jing, XIE Jun-mu-zi, WANG Feng 2019, 47 (7):  696-699.  doi: 10.11958/20181831 Abstract ( 629 )   PDF (461KB) ( 4107 )   Objective To study the effect of penta-O-galloyl-β-D-glucose (β-PGG) on the glycolysis and growth of human pancreatic cancer cells. Methods MiaPaCa-2 human pancreatic cancer cells were treated with different concentrations of β-PGG. The proliferation of MiaPaCa-2 was assessed by CCK8 method. Cytochemical kit was used to determine the product of glycolysis, lactate. Hypoxia inducible factor-1α (HIF-1α), glucose transporter 1 (GLUT1), the glycolytic enzymes of hexokinase Ⅱ (HK - Ⅱ) and phosphofructokinase (PFK) were determined by Western blot assay.Results The results of CCK8 assay showed that β-PGG inhibited the proliferation of pancreatic cancer MiaPaCa-2 cells in a dose - and time-dependent manner (P＜0.05). β - PGG also inhibited the production of lactate in both normoxic and hypoxia conditions (P＜0.05). Compared with the control group, β-PGG inhibited the expression of HIF-1α, GLUT1, HK-Ⅱ and PFK. And the inhibitory effect increased with the increased concentration of β-PGG (P＜0.05). Conclusion β-PGG can inhibit the glycolysis pathway of human pancreatic cancer cell line MiaPaCa-2 and also inhibit the proliferation of MiaPaCa-2 cells. The inhibition may be achieved by inhibiting the expressions of HIF-1α, GLUT1, HK-Ⅱ and PFK in cancer cells. Related Articles | Metrics

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Effects of pathological conditions on the expression of β3-adrenergic receptors in cardiomyocytes and cardiac fibroblasts XU Zhong-cheng, XIN Jun-zhou, WU Lei, LIU Hong-yang, WANG Li 2019, 47 (7):  700-704.  doi: 10.11958/20190274 Abstract ( 666 )   PDF (517KB) ( 3584 )   Objective To observe differences in the expression of β3-adrenergic receptor (β3-AR) under different pathological factors associated with heart failure in neonatal rat cardiac myocytes and cardiac fibroblasts. Methods The neonatal rat cardiomyocytes and cardiac fibroblasts were isolated and cultured, and both were identified by immunofluorescence. Then angiotensin Ⅱ (AngⅡ) and norepinephrine (NE) were given to stimulate the cells. And qPCR was used to detect cardiomyocyte hypertrophy-related genes (ANP, BNP and β - MHC), cardiac fibroblast-related genes(Col-Ⅰ, COL-Ⅲ) and β3-AR mRNA expression in two kinds of cells. Results Immunofluorescence identification showed that the isolated and cultured cardiomyocytes and cardiac fibroblasts were in good condition. After 48 hours of AngⅡ and NE stimulation, the expressions of ANP, BNP, β-MHC and β3-AR were increased significantly in cardiomyocytes compared with those in control group, while the expressions of Col-Ⅰ and COL-Ⅲ were increased significantly in cardiac fibroblasts compared with those of the normal control group. However, the expression of β3-AR did not change significantly.Conclusion AngⅡ and NE, pathological factors involved in the development of heart failure, mainly cause an increase in the expression of β3-AR in cardiomyocytes rather than cardiac fibroblasts. Related Articles | Metrics

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The role of mTOR-autophagy pathway on microglia phenotype in hippocampus of septic mice ZHUANG Xin-qi, JIANG Yi, YANG Man, WANG Yao-qi, LU Yue-chun, LYU Guo-yi XIE Ke-liang, YU Yong-hao 2019, 47 (7):  705-708.  doi: 10.11958/20190390 Abstract ( 767 )   PDF (750KB) ( 3787 )   Objective To investigate the role of mammalian target of rapamycin (mTOR) - autophagy pathway on microglia phenotype in hippocampus of septic mouse model. Methods Fifty-four male ICR mice were randomly allocated into three groups (n=18 for each group): Sham operation group (group Sham), cecal ligation and puncture group (group CLP)and CLP plus mTOR inhibitor rapamycin group (group CLP+Ra). Only laparotomy was performed in group Sham. Sepsis mouse model was performed by cecal ligation and puncture in group CLP. Rapamycin 1.5 mg / kg was injected intraperitoneally 6 hours before cecal ligation and puncture in group CLP+Ra. The enzyme-linked immunosorbent assay (ELISA) was conducted to assess the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-10 and transforming growth factor (TGF)-β in hippocampus. Immunofluorescence was used to calculate the number of ionized calcium binding adapter molecule (Iba) - 1 / CD86 and Iba-1 / CD206 positive microglia in hippocampal sections. The expression levels of phosphorylated mTOR (p-mTOR), microtubule associated protein 1 light chain 3Ⅱ (LC3Ⅱ) and Beclin-1 in hippocampus were detected by Western blot assay. Results Compared with group Sham, the levels of TNF-α, IL-1β, IL-10, TGF-β, the number of Iba-1 / CD86 and Iba-1 / CD206 positive microglia and the expression of p-mTOR were increased while the expression of LC3Ⅱ and Beclin-1 were decreased in group CLP (P＜0.05). Compared with group CLP, the levels of IL-10,TGF-β, the number of Iba-1/CD206 positive microglia and the expression of LC3Ⅱ and Beclin-1 were increased while the levels of TNF-α, IL-1β, the number of Iba-1/CD86 positive microglia and the expression of p-mTOR were decreased in group CLP+Ra (P＜0.05). Conclusion mTOR-autophagy pathway affects hippocampal inflammation in septic model mice by regulating microglial phenotype transformation. Related Articles | Metrics

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The effect and mechanism of transplanted MenSCs on polycystic ovary syndrome in model rats LIU Rong-hua, HE Wen-rong 2019, 47 (7):  709-712.  doi: 10.11958/20190073 Abstract ( 717 )   PDF (885KB) ( 4151 )   Objective To investigate the therapeutic effect of transplantation of menstrual stem cells (MenSCs) on rat model of polycystic ovary syndrome (PCOS). Methods A total of 45 female SD rats were divided into 3 groups. The control group was not treated. The PCOS group was intramuscularly injected with dehydroepiandrosterone (DHEA, 60 mg/ kg) to induce PCOS. The transplant group was intramuscularly injected with DHEA. After 3 weeks, the MenSCs suspension was injected through the tail vein. After 5 weeks of feeding, venous blood was taken. ELISA kit was used to detect folliclestimulating hormone (FSH), luteinising hormone (LH), estradiol (E2), progesterone (P), testosterone (T) and prolactinemia(PRL) levels. Ovarian tissues were taken for HE staining to observe ovarian morphology. qPCR was used to detect miRNA-135 expression in ovarian tissue. Western blot assay was used to detect the expression of Bax and Bcl-2 proteins in ovarian tissue. Results The FSH level was significantly decreased, and LH and T levels were significantly increased, in the PCOS group than those in the control group (P＜0.05). The FSH level was significantly increased, and LH and T levels were significantly decreased in the transplantation group than those in the PCOS group (P＜0.05). The follicle morphology was normal in the control group. Follicles with more cystic dilatation in ovary were observed in the PCOS group. The follicles with polycystic expansion were less in the transplantation group than those in the PCOS group. The expressions of miRNA-135 and Bcl-2 protein were significantly decreased in the PCOS group than those in the control group, and the expression of Bcl-2 protein was significantly higher in the PCOS group than that in the control group (P＜0.05). The expressions of miRNA-135 and Bcl-2 protein were significantly higher, the expression of Bas was significantly decreased, in the transplantation group than those of PCOS group (P＜0.05). Conclusion MenSCs has a significant improvement effect on rat PCOS, which may be achieved by up-regulating the expression of miRNA-135 and then regulating the Bax/Bcl-2 pathway. Related Articles | Metrics

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The role of myocardial autophagy in isoproterenol-induced cardiac hypertrophy SONG Dai-fu, GUO Guo-feng, LONG Hai-quan, CHEN Sheng-qiang, HUANG Jiong-hua, CHEN Xi-ming 2019, 47 (7):  713-717.  doi: 10.11958/20190662 Abstract ( 767 )   PDF (803KB) ( 3816 )   Objective To explore the effects of myocardial autophagy on isoproterenol (ISO) - induced cardiac pathological hypertrophy in rats. Methods Twenty-nine SD rats were randomly divided into three groups: subcutaneous injection of normal saline combined with intraperitoneal injection of normal saline (Con, n=9) group, subcutaneous injection of normal saline combined with intraperitoneal injection of isoproterenol (ISO, n=10) group and subcutaneous injection of rapamycin (RAP) combined with intraperitoneal injection of ISO (ISO + RAP, n=10) group. After 14-days of continuous treatment, cardiac echocardiogram for left ventricular wall thicknesses and ejection fraction (LVEF) were carried out in Con group (n=9) and ISO group (n=8). Rats were executed by intraperitoneal injection of overdoes 10% chloral hydrate. Heart to body index (HWI) was calculated by heart weight/body weight. Cardiac pathologic alternation was observed by histological sections stained with hematoxylin and eosin (HE) and Masson. The expression of p62 and the ratio of LC3 Ⅱ/Ⅰ were measured by Western blot assay. The number of autophagic vacuoles was detected by transmission electron microscopy.Results Compared with Con group, the left ventricular wall thickness and LVEF were significantly increased in ISO group (P＜0.01). Compared with Con group, the HWI was increased in ISO group and ISO+RAP group (P＜0.01). But the HWI was decreased in ISO+RAP group comparison with that of ISO group (P＜0.01). Compared with Con group, the alternation of cardiac pathological hypertrophy was marked in ISO group.Moreover, compared with ISO group, cardiac pathological hypertrophy was improved in ISO+RAP group. Compared with Con group, the ratio of LC3Ⅱ/Ⅰwas down-regulated, the protein expression of p62 was up-regulated and the number of autophagic vesicles was significantly reduced in ISO group (P＜0.01). However, compared with ISO group, the ratio of LC3Ⅱ/Ⅰwas up-regulated, the protein expression of p62 was down-regulated and the number of autophagic vesicles was significantly increased in ISO + RAP group (P＜0.01). Conclusion The up-regulation of myocardial autophagy can reverse pathological hypertrophy induced by ISO. Related Articles | Metrics

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Local recurrence risk and survival analysis of breast cancer patients with T1-2 and one to three positive axillary lymph nodes without postmastectomy radiotherapy LOU Li-ping, SHI Zhen-dong, LIU Jing-jing, YANG Xiao-nan, ZHANG Jin 2019, 47 (7):  718-723.  doi: 10.11958/20190345 Abstract ( 1066 )   PDF (438KB) ( 4329 )   Objective To investigate the relationship between molecular subtype and local recurrence and prognosis in patients with early-stage breast cancer and 1-3 positive axillary lymph nodes (ALNs), and to improve the individualized indications for postmastectomy radiotherapy (PMRT). Methods Clinical data of 489 patients with pT1-2N1M0 (1-3 positive ALNs) who did not receive PMRT in our hospital from 2009 to 2012 were analyzed retrospectively. The KaplanMeier method was used to calculate local recurrence rate (LRR), disease-free survival rate (DFS) and overall survival rate(OS). The Log-rank test and multivariate Cox analysis were used to determine the risk of LRR associated with different molecular types and to determine the clinicopathological factors associated with prognosis. Results In 489 breast cancer patients with pT1-2N1M0 and without PMRT, there were 229 patients with Luminal A subtype, 196 with Luminal B subtype(HER-2 negative) and 64 with triple-negative subtype, respectively. The median follow-up time was 75 months (range from 5-115 months). The molecular typing analysis showed that the 5-year LRR of the triple-negative subtype was significantly higher than that of the non-triple-negative subtype (P＜0.05), and the 5-year DFS and the 5-year OS were significantly lower than the non-triple-negative subtypes (P＜0.05). There were no significant differences in 5-year LRR, DFS, and OS between Luminal A and Luminal B (HER-2 negative) in the non-triple-negative subtypes (P＞0.05 each). Multivariate Cox regression analysis showed that age ≤ 35 years, pT2 stage and triple-negative subtypes were independent poor prognostic factors for LRR. Conclusion Molecular subtyping is helpful for the individualized evaluation of LRR in pT1-2N1M0 breast cancer patients with 1-3 positive ALNs. We recommend PMRT to improve the prognosis for patients with age ≤ 35 years, pT2 stage and triple-negative subtypes. Related Articles | Metrics

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Changes of cognitive function and its associated risk factors in hemodialysis patients TIAN Ru, GUO Yi-dan, LUO Yang 2019, 47 (7):  723-726.  doi: 10.11958/20190258 Abstract ( 806 )   PDF (336KB) ( 3901 )   Objective To analyze the characteristics of cognitive function changes and its associated risk factors in hemodialysis patients. Methods Baseline data of hemodialysis patients and the results of two Montreal Cognitive Assessment Scale (MoCA) tests were collected. The characteristics of cognitive function changes in patients in two years were analyzed, and the associated risk factors of cognitive function changes were analyzed by multiple linear regression method.Results A total of 103 patients were included in the study (67.0%). It was found that age, dialysis age and the prevalence rates of stroke and heart disease were higher in cognitive impairment group than those in normal group. And hemoglobin level, education and the scores of MoCA were significantly lower in cognitive impairment group than those in normal group(P＜0.05). After two years, the scores of MoCA, visual space, executive function and delayed recall were significantly lower than those of baseline (P＜0.05). Multivariate linear regression analysis showed that changes of MoCA were affected by age, education level, history of heart disease and dialysis age; changes of visual space and executive function were affected by age, education level and history of heart disease; and changes of delayed recall were affected by age, education level and dialysis age (all P＜0.05). Conclusion Cognitive function shows a significant downward trend during a two-year period in hemodialysis patients, which are mainly in the area of visual space, executive function and delayed recall. Older age, lower education, history of heart disease and longer dialysis age are risk factors for cognitive decline in these patients. Related Articles | Metrics

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Changes and significance of OX40 mRNA and Th1 in peripheral blood of acute rejection recipients after renal transplantation ZUO Ya-wei, MUO Chun-bai, LI Guang, GENG Jie, WANG Yu-liang 2019, 47 (7):  727-731.  doi: 10.11958/20190386 Abstract ( 587 )   PDF (596KB) ( 3477 )   Objective To investigate the significance of determining OX40 mRNA and Th1, Th2 of CD3+CD8- T helper lymphocytes in peripheral blood for evaluating renal acute rejection. Methods A total of 40 recipients with renal transplantation and 20 healthy volunteers were enrolled and divided into acute rejection group (n=20) and stable renal allograft function group (n=20) according to the inclusion criteria. The levels of OX40 mRNA in peripheral blood mononuclear cells (PBMCs) were measured by TaqMan real-time reverse transcriptase-polymerase chain reaction (RTPCR) assays. The expression percentages of IFN-γ and IL-4 of CD3+CD8- T lymphocytes (Th1 and Th2) in peripheral blood were determined with flow cytometry. Receiver operating characteristic (ROC) curve was used to evaluate the prediction value. Results The level of OX40 mRNA in PBMCs significantly elevated in acute rejection group compared with that in stable renal allograft function group and normal control group (P＜0.01). There were no significant differences in OX40 mRNA levels between stable renal allograft function group and normal control group (P＞0.05). The expression percentage of Th1 in peripheral blood was significantly higher in acute rejection group compared with that in stable renal allograft function group and normal control group (P＜0.01). No significant difference was found in the expression percentage of Th2 in peripheral blood between the three groups (P＞0.05). The area under ROC curve (AUC) in the diagnosis of acute rejection was 0.903 (95% CI 0.825-0.980) for OX40 mRNA. The AUC of Th1 was 0.731 (95%CI：0.587-0.874). The AUC of combined diagnosis was 0.930 (95% CI: 0.859-1.001). Conclusion The monitoring of OX40 mRNA and Th1 after renal transplantation can predict acute renal allograft rejection. Related Articles | Metrics

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Study on the sST2/IL-33 and Th17/Treg related cytokine in patients with Primary Immune Thrombocytopenia WANG Xiu-juan, SUN Ming-ling, LIU Ying, WANG Lei, WANG Xin-you, LIU Yang, ZHAO Fang, QIN Yu-ting, GUO Xin-hong 2019, 47 (7):  732-734.  doi: 10.11958/20182195 Abstract ( 691 )   PDF (319KB) ( 3495 )   Objective To observe the changes of expression levels of serum soluble ST2 (sST2), interleukin-33 (IL-33), and their regulation effects on Th17 / Treg cell immune balance in primary immune thrombocytopenia (ITP), and to investigate the possible immune mechanism of sST2 and IL-33 in ITP. Methods A total of 30 patients with ITP were regarded as ITP group, and 20 healthy subjects were used as healthy control group. ELISA method was used to detect the serum levels of sST2, IL-33, IL-17 and TGF-β in two groups. Results Compared with the normal control group, the serum levels of sST2 and IL-17 were increased (P＜0.01), the serum level of TGF-β was decreased in ITP group (P＜0.01). There was no significant change in serum level of IL-33 between the two groups (P＞0.05). The correlation analysis showed that sST2 was negatively correlated with TGF-β in ITP group (r=-0.471, P＜0.01), and there was no correlation in sST2 and IL-17 (r=0.189, P＞0.05). Conclusion The increased expression of sST2 and the imbalance of Th17/Treg may be one of the key immunomechanisms of ITP. Related Articles | Metrics

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The value of inspiratory muscle training as a rehabilitation treatment in patients with chronic obstructive pulmonary disease ZHENG Jiang-nan, XIAO Ying, ZOU Zhao-hua, WU Qiao-zhen, HU Xiao-yun, ZHANG Jian-feng, DU Huan 2019, 47 (7):  735-738.  doi: 10.11958/20181904 Abstract ( 828 )   PDF (359KB) ( 4744 )   Objective To investigate the effect of inspiratory muscle training (IMT) as a rehabilitation treatment for patients suffering from moderate to severe chronic obstructive pulmonary disease (COPD). Methods Fifty COPD patients in stable stage hospitalized in the First People’s Hospital of Wu Jiang District, Suzhou from January 2014 to December 2015 were included and randomly divided into control group and IMT group. Patients in the control group inhaled bronchodilators or glucocorticoids, whereas those in IMT group took inspiratory muscle training in addition to inhaling bronchodilators or glucocorticoids. All patients were followed up for 12 months. Values of maximum inspiratory pressure (PImax), 6-minute walking distance (6MWD), COPD Assessment Test (CAT) score were analyzed. The number of hospitalizations within twelve months was also recorded in two groups of patients. Results After 12-month fellow-up, it was found that PImax was decreased［(80.12 ± 17.94) cmH2O vs. (78.28 ± 17.98) cmH2O, P＜0.05)］, 6MWD was shortened［(283.96 ± 22.67) m vs.(278.12 ± 21.12) m, P＜0.05］and FEV1 was decreased［(1.32 ± 0.25) L vs. (1.27 ± 0.24) L, P＜0.05］in control group.Compared with control group, PImax increased significantly in IMT group［(78.28±17.98) cmH2O vs. (95.48±19.36) cmH2O,P＜0.05］, 6MWD was also increased［(278.12 ± 21.12) m vs. (320.32 ± 28.71) m, P＜0.05)］. While the CAT score was significantly reduced in patients of IMT group (15.36±5.20 vs. 11.76±2.79, P＜0.05). The hospitalizations caused by acute exacerbation were significantly decreased (1.40±0.91 vs. 0.68±0.63, P＜0.05), but there was no significant improvement in pulmonary function (P＞0.05). Conclusion Long-term IMT can improve the activity tolerance and quality of life in COPD patients, and reduce hospitalizations, bring benefits to the survival of COPD patients. Related Articles | Metrics

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The clinical characteristics of 74 cases of severe fever with thrombocytopenia syndrome HE Zhi-quan, YUAN Chun, CUI Ning, HUANG Xue-yong, DU Yan-hua, LI Yi, SU Jia, XU Bian-li, GUO Wan-shen 2019, 47 (7):  738-741.  doi: 10.11958/20190002 Abstract ( 706 )   PDF (367KB) ( 4180 )   Abstract: Objective To analyze the clinical characteristics of severe fever with thrombocytopenia syndrome (SFTS)and provide more theoretical basis for the treatment. Methods A retrospective analysis was performed on 74 confirmed cases collected from the 154 Hospital, Chinese People’s Liberation Army, Xinyang City, Henan Province from August 2017 to August 2018. The patients were divided into mild group (58 cases) and severe group (16 cases), and the differences in the clinical features and laboratory test were compared between the two groups, including white blood count (WBC), platelet (PLT), aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatine kinase (CK), lactate dehydrogenase (LDH)and blood urea nitrogen (BUN). Multiple Logistic regression analysis was conducted to explore the risk factors in SFTS patients. Results The epidemiological data showed that most of patients were older farmers. Clinical features of SFTS patients contained fever, loss of appetite, fatigue, nausea, muscle pain and lymphadenectasis. The incidences of the loss of appetite, diarrhea, unconsciousness, chest distress and asthma were significantly higher in severe group than those in mild group (P＜0.05). The laboratory test showed that CK and BUN levels were significantly higher in severe group than those in mild group (P＜0.05), but there were no significant differences in the other indictors between the two groups. Logistic regression analysis showed that loss of appetite (OR=5.207, 95%CI：1.039-26.082) and CK≥200 U/L (OR=4.446，95%CI：1.231-16.058) were risk factors leading to the progression of disease from mild to severe. Conclusion The loss of appetite and increased level of CK are risk factors for the aggravation of the disease. These factors should be controlled in order to reduce mortality. Related Articles | Metrics

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Analysis of influencing factors of abdominal aortic balloon occlusion technique on intraoperative hyperlactacidemia in patients selected for surgical excision of pelvic and sacrum tumors XU Jun-jun, GAO Ling-ling, FENG Yi 2019, 47 (7):  742-746.  doi: 10.11958/20182201 Abstract ( 774 )   PDF (351KB) ( 5294 )   Objective To analyze relevant factors for intraoperative hyperlactacidemia in patients selected for surgical excision of pelvic and sacrum tumors assisted by abdominal aortic balloon occlusion and explore the relationship between intraoperative hyperlactacidemia and postoperative complications. Methods The clinical data of 341 patients underwent surgical excision of pelvic and sacrum tumors assisted by abdominal aortic balloon occlusion in Peking University People’s Hospital from May 3, 2016 to May 3, 2018 were retrospectively analyzed. Logistic regression analysis was performed on factors including age, gender, American Association of Anesthesiologists (ASA) classification, body mass index (BMI),preoperative hemoglobin content, preoperative albumin content, surgery time, blood loss, urine output, total occlusion time of aortic balloon and lactate levels. Data of post-operative cardiac，pulmonary and neurological system complications and surgical wound infection were also collected. The patients were divided into hyperlactacidemia group (n=71) and control group (n=270) according to whether hyperlactacidemia developed during the operation. The incidences of postoperative complications were also compared between the two groups. Results Seventy-one patients (71 / 341, 20.82%) developed intraoperative hyperlactacidemia. Multivariate Logistic regression analysis showed that total occlusion time of aortic balloon(＞60 min) and blood loss(＞4 000 mL) were the independent risk factors of intraoperative hyperlactacidemia. The incidence of post-operative pulmonary complications was significantly higher in patients with intraoperative hyperlactacidemia than that in patients without intraoperative hyperlactacidemia (P＜0.05). There were no significant differences in incidences of post-operative cardiac and neurological system complications and the incidence of postoperative wound infection between the two groups (P＞0.05). Conclusion Patients with total occlusion time of aortic balloon more than 60 min and intraoperative blood loss more than 4 000 mL are prone to intraoperative hyperlactacidemia. Patients with intraoperative hyperlactacidemia are associated with higher incidence of post-operative pulmonary complications than patients without intraoperative hyperlactacidemia. Special attention should be given to these patients. Related Articles | Metrics

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Changes and clinical significance of serum IL-37 in patients with hepatitis B virus-related acute-on-chronic liver failure SHEN Min, WU Kun-liang, HE Jing, ZHOU Yun, ZHOU Gui-zhong 2019, 47 (7):  746-749.  doi: 10.11958/20182229 Abstract ( 577 )   PDF (367KB) ( 3855 )   Objective To investigate the expression of interleukin-37 (IL-37) in patients with hepatitis B virusrelated acute-on-chronic liver failure (HBV-ACLF) and its clinical significances. Methods Eighty-one patients with HBV-ACLF admitted to our hospital from June 2015 to June 2017 were enrolled as study subjects (HBV-ACLB group), and 30 patients with chronic hepatitis B (CHB) were used as CHB group and 30 healthy people with physical examination were used as control group. In HBV-ACLB group there were 25 patients at early stage, 32 cases at middle stage and 24 cases at advanced stage at admission. Markers of liver function and serum levels of IL-37 were compared between three groups of patients. Pearson or Spearman correlation analysis were used to analyze the correlation between IL-37 level and markers of liver function, and the prognosis of HBV-ACLF. Results There was significantly higher serum level of IL-37 in HBVACLF group than that of CHB and control groups (P＜0.05). Among the patients with HBV-ACLF, those in the advanced stage had significantly higher levels of IL-37 than those in the patients with middle stage and early stage,and those in the middle stage had significantly higher levels of IL-37 than those in the early stage (P＜0.05). There were positive correlation between IL-37, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in HBV-ACLF group (rs values were 0.52 and 0.72, respectively). Among 81 cases of HBV-ACLF, 22 cases acquired infection and 39 cases died during the 2-month follow-up period. The serum levels of IL-37 were significantly higher in the infected cases and death cases than those in the non-infected cases and survival cases (P＜0.05). Conclusion The serum level of IL-37 is associated with the severity of HBV-ACLF, and which can be used as an important prognostic marker for ACLF. Related Articles | Metrics

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The clinical observation of laparoscopic splenectomy for immune thrombocytopenia with platelet counts ＜ 10×109/L CAO Lei, ZHENG Kun-ming, WANG Hong-lei, LIAO Mao-mao, XUE Feng, ZHANG Lei, ZHAO Yong-jie, JIANG Tao 2019, 47 (7):  750-753.  doi: 10.11958/20190587 Abstract ( 808 )   PDF (551KB) ( 3599 )   Objective To investigate the feasibility and clinical efficacy of laparoscopic splenectomy (LS) for immune thrombocytopenic purpura (ITP) with platelet counts lower than 10×109/L. Methods The clinical data of 17 patients of ITP with platelet (PLT) count lower than 10×109/L, who underwent LS between March 2006 to June 2016 were retrospectively analyzed．There were 8 male cases and 9 female cases in these patients. The mean age was (42.18±15.70) years. Patients were found PLT count < 10×109/L 1 day before surgery, the median course of disease was 2 (1, 5.5) years, and 6 patients were found splenomegaly. Results All 17 patients were successfully underwent LS. The operation time was (156.76±17.90) minutes, median blood loss volume of intraoperative blood loss was 50 (20, 175) mL, and the average hospitalization after surgery was (7.47±1.94) d. The preoperative coagulation functions were normal in all 17 cases. No uncontrol bleeding and postoperative complications occurred in intraoperative and postoperative time. Twelve cases were effectiveness in the short term after LS operation, including 11 cases of complete response and 1 case of partial response. The postoperative effective rate of PLT≥100×109/L was significantly higher than that of PLT < 100×109/L at postoperative 1 d (P＜0.05). There were no significant differences in the postoperative effective rate among different ages, genders, course of disease, whether accompanied by splenic and preoperative immunoglobulin blocking therapy and other factors. Conclusion LS is safe,effective and feasible for the treatment of refractory ITP. For patient with preoperative PLT＜10×109/L, and PLT one day after operation could be an indicator to predict the short-term efficacy of LS for the treatment of ITP. Related Articles | Metrics

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Congenital arhinia of fetus: a case report ZHANG Qiang, PENG Jin-zhi, LI Quan, WANG Shao-jun 2019, 47 (7):  754-757.  doi: 10.11958/20190368 Abstract ( 661 )   PDF (494KB) ( 3701 )   Congenital arhinia is a fetal malformation with unknown etiology, unclear pathogenesis and extremely rare clinically. It often accompanied by other organ malformations, which affects the outcome of perinatal infants in varying degrees. Therefore, a skilled neonatal resuscitation team is needed during delivery. Most fetuses with arhinia can live normally after birth. At present, surgery is still the main therapeutic method for arhinia, and stem cell therapy is expected to become a new treatment method. This paper reported a case of arhinia of fetus admitted to our hospital, and the relevant literature was reviewed. The purpose of this study is to raise awareness of the disease, reduce missed diagnosis and misdiagnosis, provide necessary prenatal counseling and help for patients and their families, and give sound advice for neonatal care, feeding and treatment. Related Articles | Metrics

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Hemorrhage of renal hamartoma in late pregnancy: a case report and literature review LU Wen-bao, LIU Xiao-liang, LENG Yuan-jing, HUANG Yu-qing, ZHANG Liang 2019, 47 (7):  758-761.  doi: 10.11958/20182139 Abstract ( 735 )   PDF (531KB) ( 3730 )   Renal hamartoma is one of the most common renal tumors. Most of them are benign and tend to rupture and bleed when they are large. However, there are few cases of the late pregnancy complicated with renal hamartoma hemorrhage,and pathological evidence shows malignant potential. This article reviews and analyses the diagnosis and treatment data of one case of ruptured renal hamartoma in late pregnancy, and summarizes the literature to discuss the treatment of renal hamartoma hemorrhage. Related Articles | Metrics

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Application value of ultrasound strain elastography in the diagnosis of benign thyroid nodule fibrosis WANG Ming-yang, TANG Ying, WANG Jin, YU Hui-min, YANG Mu-lei 2019, 47 (7):  761-764.  doi: 10.11958/20190052 Abstract ( 714 )   PDF (673KB) ( 4110 )   Objective To investigate the application value of ultrasound strain elastography in the diagnosis of benign thyroid nodules with fibrosis. Methods A total of 155 benign thyroid nodules confirmed by pathology were selected.Ultrasound strain elastography was performed before operation. According to the pathological results, they were divided into fibrosis group (n=125) and non-fibrosis group (n=30). The ratio of strain was compared between the two groups. The sensitivity and specificity of the strain ratio in the diagnosis of benign thyroid nodules with fibrosis were calculated by using pathological results as the gold standard. Results The strain ratio was significantly higher in the fibrosis group [3.14 (1.14,7.05)] than that in the non-fibrosis group [1.41 (1.02, 2.35), P＜0.05]. The ROC curve showed that the stain ratio＞2.62 may indicate the presence of fibrosis in the benign thyroid nodule. The area under the curve of the strain ratio was 0.664 (0.548,0.780), the sensitivity was 56.7%, and the specificity was 77.6%. Conclusion The strain ratio of ultrasound elastography has a certain value in the diagnosis of benign thyroid nodules with fibrosis. Related Articles | Metrics

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Vedolizumab for treatment of inflammatory bowel disease: research progress of clinical applications and mechanisms WANG Hui-qin, LIANG Zhao-liang, LIU Geng-feng, LYU Xiao-dan, ZHAN Ling-ling, LYU Xiao-ping 2019, 47 (7):  765-770.  doi: 10.11958/20181974 Abstract ( 1245 )   PDF (377KB) ( 3896 )   Inflammatory bowel disease (IBD) is a kind of chronic, relapsing inflammatory disease of the gastrointestinal tract with unknown etiology. Its treatment is still dominated by traditional medicine. Due to the obvious adverse reactions of these traditional drugs, a variety of biological agents have been derived in recent years, such as anti-tumor necrosis factor(TNF) biological agents, integrin antagonists. As a humanized anti- α4β7 integrin monoclonal antibody, vedolizumab can selectively block trafficking of memory T cells to inflamed intestinal tissues by inhibiting the interaction of α4β7 integrin with mucosal address cell adhesion molecule-1, reducing intestinal inflammation. Therefore, it is mainly used in IBD patients who are failed to respond to the anti-TNF biological agents. This review will discuss the mechanism, clinical efficacy and safety of vedolizumab in the treatment of IBD. Related Articles | Metrics

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Diagnosis of autoimmune pancreatitis SHANG Hai-tao, LI Zhong-lian 2019, 47 (7):  771-776.  doi: 10.11958/20191155 Abstract ( 983 )   PDF (383KB) ( 4250 )   Autoimmune pancreatitis (AIP) is a distinct form of chronic pancreatitis, which is clinically divided into two types (type Ⅰ and type Ⅱ) according to histology. Type Ⅰ is the main disease in China, which is a part of the IgG4-related disease characterized clinically by obstructive jaundice or a pancreatic mass, histologically by numerous lymphoplasmacytic infiltrate and chronic fibrosis. Although the International Association of Pancreatology (IAP) has developed diagnostic criteria for its imaging, serology, histology and hormone therapy responses, the clinical features of autoimmune pancreatitis are occult and varied, the diagnosis of autoimmune pancreatitis remains challenging, particularly in patients with pancreatic masses, which need to be alert to the risk of the pancreatic cancer. This review is to summarize the literature from the perspective of image and histology, focusing on the diagnosis and risk of misdiagnosis of this complex disease, in order to improve the diagnosis of autoimmune pancreatitis. Related Articles | Metrics

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The research progress of radiomics in hepatocellular carcinoma GUO Dong-hui, JI Qian, WANG Hong-hai, ZHENG Hong 2019, 47 (7):  777-780.  doi: 10.11958/20190034 Abstract ( 872 )   PDF (341KB) ( 3533 )   Radiomics, the high-throughput mining of quantitative image features from standard-of-care medical imaging that enables data to be extracted and applied within clinical-decision support systems to improve the predictive accuracy of diagnosis and prognosis, is gaining importance in cancer research. Radiomics utilizes complex image analysis capabilities of computer and massive amounts of medical imaging data, providing a powerful tool in modern medicine. This article briefly introduces the concept of radiomics and its research progress in the field of hepatocellular carcinoma. Related Articles | Metrics

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Advances in diagnosis and treatment of acute exacerbation of IPF after lung cancer surgery ZHOU Fang, LU Xi-ke, ZHANG Xun, WANG Zheng, LI Yue-chuan 2019, 47 (7):  781-784.  doi: 10.11958/20190733 Abstract ( 1230 )   PDF (337KB) ( 4776 )   Idiopathic pulmonary fibrosis (IPF) is a respiratory disease characterized by slow progression. It is a special type of interstitial pneumonia. Its pathological changes are confined to lungs. It is common in the elderly. However some cases occur with acute exacerbation that manifested by sudden deterioration of the original pulmonary lesions, fatal respiratory failure and new pulmonary shadow and diffuse alveolar pathological changes. This acute exacerbation of IPF can occur in patients with primary lung cancer after lung resection, usually with an acute or subacute onset. The etiology of acute exacerbation of pulmonary fibrosis after lung cancer surgery is still unclear. It is usually considered to be related to pulmonary injury of unknown causes. Because of its hidden onset, rapid progression, high mortality and lack of routine preventive and therapeutic measures in clinic, it has become a difficult problem for clinicians. This article reviews the etiology, risk factors, predictive indicators, prophylaxis, treatment and prognosis of acute exacerbation of IPF after lung cancer surgery. Related Articles | Metrics