1.Expression of tumor heterogeneity-related genes in esophageal squamous cell carcinoma and its predictive value for chemotherapy sensitivity
Wenhui WANG ; Yuanling QI ; Haipeng REN ; Qian XU
Chinese Journal of Geriatrics 2015;34(6):635-637
Objective To investigate the expression of excision repair cross-complementation group 1 (ERCC1) and ribonucleotide reductase subunit M1 (RRM1) in patients with esophageal squamous cell carcinoma,and the relationship between the expression of ERCC1 and RRM1 genes and chemotherapy sensitivity.Methods A total of 77 patients with esophageal squamous cell carcinoma admitted in hospital from Jan.2008 to Dec.2012 were recruited.ERCC1 and RRM1 mRNA levels in the cancerous tissue were detected by real-time fluorescent quantitative RT PCR.The relationship between short-term effects of chemotherapy and ERCC1 and RRM1 mRNA levels was analyzed.Results Levels of mRNA in stages Ⅰ a,Ⅰ b,Ⅱ a,and Ⅱ b were (0.578±0.081),(0.560±0.084),(0.521±0.080),(0.464±0.091) for ERCC1 and(0.511±0.089),(0.539± 0.086),(0.584±0.092),(0.637±0.101)for RRM1,respectively.As the clinical stage advanced,the ERCC1 mRNA level declined and the RRM1 mRNA level increased (t=2.679 and 2.952,P =0.034 and 0.025,respectively).Levels of mRNA in patients with complete remission,partial remission,stable disease and progressive disease were (0.487 ± 0.097,0.511 ± 0.095,0.552 ± 0.086,0.568 ± 0.088) for ERCC1 and(0.504±0.091,0.544±0.095,0.595±0.093,0.616±0.097) for RRM1,respectively.The clinical effect of chemotherapy was negatively correlated with mRNA expression of ERCC1 and RRM1 (r=0.567,P=0.032).Conclusions Levels of ERCC1 and RRM1 mRNA expression are correlated with the staging of esophageal squamous cell carcinoma and chemotherapy sensitivity,and can be used as a predictive parameter for chemotherapy sensitivity in patients with esophageal squamous cell carcinoma.
2.Glutamate Involved in Rats with Spinal Cord Injury Induced by Dynorphin A (1-17)
Fuchun LI ; Pingping ZUO ; Wenhui HU ; Na LIU ; Minfeng REN
Chinese Journal of Rehabilitation Theory and Practice 1997;3(1):6-9
Intrathecal (I. T.)administration of K opioid dynorphin A (1-17) is used as a model of neurological dysfunction which lnvolved in spinal cord injury and secondary affection according to several previous reports. 5-amlno-2-phosphoveroleric acid (APV ), an NMDA receptor antagonist, can significantlly prevent the hindlimb paralysis in rats produced by I. T. dynorPhin A (1-17). To further investigate the mechanisms, we establis11 a nlodel of [3H]L,-Glu release in rats spinal slices influenced by dynorphin A (dynA ) (1-17). [3H]L-Glu release evoked by high [K+] (5Ommol/L,)is a Ca2+-dependent process. DynA (1-17) slgnificantly inhibited [3H]L,-Glu release at 1O-8mol/L,, but very significantly enhanced [3H]L-Glu release at 10-6 mol /L. The synthetic k agonist U50, 488H, which has no neurotoxic effect, inhibited [3H]L-Glu release at both high and low concentrations and did not have any enhancing effect. The results suggest that the analgesic effect of dynA (1-17) at physiological dosage may be rnediated by presynaptic K opioid receptor through the inhibition of Ca2+ influx and L-Glu release;but dynA (1-l7)enhanced L-Glu release through a non-opioid pathway and induced hindlimb paralysis at high neurotoxic dosage.
3.Mechanism of polypyrimidine tract-binding protein 1 on the proliferation and metastasis of gastric cancer cells
Yarui LI ; Mudan REN ; Guifang LU ; Xinlan LU ; Qian ZHAO ; Dan GUO ; Wenhui MA ; Shuixiang HE
Chinese Journal of Digestion 2021;41(2):100-106
Objective:To explore the expression of polypyrimidine tract-binding protein 1 (PTBP1) in gastric cancer (GC) tissues and GC cell lines, and the role of PTBP1 in the proliferation and metastasis of GC cells.Methods:From January to June in 2019 at The First Affiliated Hospital of Xi′an Jiaotong University, the cancer tissues and corresponding para-cancer tissues of GC patients underwent surgical resection were collected. The Kaplan-Meier Plotter database was used to analyze the survival of GC patients. The expression of PTBP1 was down-regulated by transfecting small interfering RNA (siRNA) in human GC cell lines SGC7901 and AGS with relatively high expression of PTBP1. The cells were divided into blank control group, negative control group, and PTBP1 knockdown group. The expression of PTBP1 at mRNA and protein level were detected by real-time fluorescence quantification polymerase chain reaction (RT-qPCR) and Western blotting. At 24, 48, 72 and 96-hour after transfection, the effect of PTBP1 on the proliferation of GC cells was observed by 3-(4, 5 dimethylthiazol)-2, 5 diphenyltetrazolium bromide (MTT) experiment. The changes of invasion and migration of GC cells after down-regulation of PTBP1 were detected by transwell assay. The expression changes of epithelial-mesenchymal transition (EMT) markers E-cadherin, N-cadherin and vimentin after down-regulation of PTBP1 in GC cells were determined by Western blotting. Indenpendent samples t test, analysis of variance and rank sum test were used for statistical analysis. Results:The Kaplan-Meier Plotter prognostic analysis showed that the overall survival of GC patients with high PTBP1 expression was shorter than that of GC patients with low PTBP1 expression (9.2 months, 6.2 months to 17.2 months vs. 19.0 months, 14.5 months to 28.4 months), and the difference was statistically significant ( Z=5.31, P<0.05). The results of RT-qPCR showed that in GC cell lines SGC7901 and AGS, the expression of PTBP1 at mRNA level of PTBP1 knockdown group was lower than that of blank control group and negative control group (SGC7901: 0.78±0.11 vs.3.10±0.19 and 2.99±0.23; AGS: 0.80±0.09 vs. 3.55±0.24 and 3.50±0.18), and the differences were statistically significant ( tSGC7901=10.57 and 8.08, tAGS=10.91 and 13.42; all P<0.01). The results of Western blotting indicated that in GC cell lines SGC7901 and AGS, the expression of PTBP1 at protein level of PTBP1 knockdown group was lower than those of blank control group and negative control group (SGC7901: 0.38±0.04 vs. 1.42±0.05 and 1.35±0.09; AGS: 0.17±0.02 vs. 1.52±0.08 and 1.38±0.45), and the differences were statistically significant ( tSGC7901=15.94 and 10.57, tAGS=16.60 and 20.80; all P<0.01). The results of MTT showed that at 48, 72 and 96-hour after transfection the absorbance values of PTBP1 knockdown group decreased by 0.25±0.01, 0.38±0.02, and 0.84±0.04 as compared with those of negative control group, and the decrease was the most significant at 96-hour after transfection, and the differences were statistically significant ( t=10.21、14.32, both P<0.01). The results of transwell experiment demonstrated that the number of invasion and migration cells of PTBP1 knockdown group were both less than that of the blank control group and the negative control group (SGC7901: 42.00±5.91 vs. 116.40±10.23 and 114.40±10.43; 39.60±6.77 vs. 125.80±11.51 and 122.40±5.90; AGS: 40.20±7.25 vs. 115.60±14.63 and 117.40±9.12; 36.00±5.20 vs. 122.40±12.10 and 125.40±12.74), and the differences were statistically significant ( tSGC7901=14.07, 13.50, 14.43 and 20.62; tAGS=10.27, 14.75, 14.68 and 16.76; all P<0.01). The results of Western blotting showed that the expression of E-cadherin of PTBP1 knockdown group was higher than that of the blank control group and the negative control group (SGC7901: 1.42±0.05 vs. 0.53±0.05 and 0.57±0.03; AGS: 1.34±0.04 vs. 0.54±0.03 and 0.61±0.01), however the expression levels of N-cadherin and vimentin were both lower than those of the blank control group and the negative control group (SGC7901: 0.50±0.03 vs. 1.64±0.05 and 1.46±0.07; 0.32±0.07 vs. 1.42±0.07 and 1.33±0.07; AGS: 0.37±0.06 vs. 1.47±0.04 and 1.36±0.04; 0.41±0.04 vs. 1.53±0.06 and 1.37±0.04), and the differences were statistically significant ( tSGC7901=11.63, 13.19, 18.83, 11.68, 11.43 and 10.43; tAGS= 15.02, 16.23, 14.67, 12.97, 14.45 and 17.18; all P<0.01). Conclusions:The expression levels of PTBP1 increase in GC tissues and cells, which may be involved in regulating the proliferation, metastasis and EMT of GC cells.
4.Analysis of HIV Infection and New Infections Detection of AIDS Sentinel Surveillance Focus Groups in Shaanxi Province 2009~2014
Qiang REN ; Wenhui CHANG ; Mengyan ZHANG ; Ting HU ; Hua LI ; Yulin FU ; Yangfan ZOU ; Lifang DONG
Journal of Modern Laboratory Medicine 2015;(3):56-59
Objective To learn 2009~2014 Shaanxi Province sentinel surveillance six classes of HIV infection focus groups, and estimates of HIV-1 new infection.Methods Used enzyme-linked immunosorbent assay (ELISA)and Western blot (WB)experiments for the 2009~2014 HIV sentinel surveillance Shaanxi Province Category 6 focus groups conducted a total of 77 778 HIV antibody screening and confirmatory testing estimates of HIV-1 new infection.Results 2009~2014 men who had sex with men and people with HIV infection rate were 3.75%,8.77%,3.50%,5.00%,6.20% and 5.75%,and a slow upward trend;HIV-1 new infection were 5.04%,8.96%,5.01%,5.95%,4.68% and 6.39%,the overall downward trend. Young students,drug addicts,sex workers,pregnant women,and male STD clinic attenders five people with HIV infection and HIV-1 new infection were emerging to remain low.But male STD clinic attenders of HIV infection and HIV-1 new in-fection was emerging slowly rising trend.Conclusion Shaanxi MSM HIV infection and HIV-1 new infection were high,but HIV-1 new infection had decreased slowly.Emerging trend should continue to increase the population of the intervention ef-forts.The infection rate in other monitoring population was relatively low but a few people on the rise,the need to take the necessary coping methods.
5.Study on Quality Control of Zhiqikang Capsules
Xin LIAO ; Yue WANG ; Wenhui QIAN ; Lu HUANG ; Hua SU ; Haixiang REN
China Pharmacist 2014;(2):221-224
Objective:To establish the quality control of Zhiqikang capsules. Methods:TLC was used to identify Gastrodia tuder halimasch, rhubarb and Astragalus mongholicus in the preparations. A spectrophotometry method with 3, 5-dinitrosalicylic acid (DNS) was used to measure the polysaccharide content in Zhiqikang capsules. A spectrophotometry method with Forint phenol method ( Low-ry) was used to measure the peptide content in the capsules. Results:The linear range of polysaccharide was obtained between 6. 412 and 32. 060μg·ml-1(r=0. 999 5), the average recovery was 95. 86% and RSD was 0. 86%. The linear range of peptide was ob-tained between 0.059 7 and 0.298 4 mg·ml-1(r=0.999 0), the average recovery was 100.3% and RSD was 1.88%(n=6). Conclusion:The assay method is simple and accurate in the quality control of the preparations.
6.Protection of INS-1 cells from STZ injury by gastrodine
Lei ZHANG ; Hongmei CHEN ; Xun LI ; Baoli ZHANG ; Wenhui REN ; Zhongping LI ; Tianzhen ZHENG ; Yumin LI
Journal of Endocrine Surgery 2011;05(4):222-224
ObjectiveTo study effects of gastrodine (GAS) on insulinoma (INS-1) cells and the protection of INS-1 ceils from steptozotocin (STZ) injury by gaatrodine. MethodsThe experiment was carried out in 5 groups: normal control group ( NC), GAS group (GAS), streptozotocin group (STZ), GAS protection group ( GAS +STZ) and GAS repair group (STZ +GAS). INS-1 cells were cultured, the cell viability was determined by tetrazolium (MTT) assay, insulin concentration was detected by radioimmunoassay, and malondialdehyde (MDA)concentration and total antioxidant capacity (T-AOC) of the culture medium were measured by colorimetry. Results GAS promoted insulin release of INS-1 cells (P<0.05, P<O.01). Low-concentration GAS could increase viability of INS-1 cells ( P < 0.01 ). GAS could increase viability of the injured INS-1 cells (P < 0.01 ). High concentration GAS contributed in repair of INS-1 cells injured by STZ and promoted insulin serection ( P < 0.01 ). GAScould decrease MDA concentration (P <0.01 ) and significantly increase T-AOC capacity of INS-1 cells injured by STZ (P <0.01 ). ConclusionsGAS can increase INS-1 viability, promote insulin secretion of INS-1 cells, alleviate INS-1 cells injury caused by STZ, and strengthen the antioxidant capacity of INS-1 cells injured by STZ.
7.Expression of urotensinⅡ receptor GPR14 in aorta of apoE knockout mice
Zhijian WANG ; Wenhui DING ; Libin SHI ; Lei MENG ; Ziwen REN ; Dingfang PU ; Yonggang ZHANG ; Chaoshu TANG
Chinese Journal of Pathophysiology 2000;0(11):-
] AIM: To investigate the expression of the urotensin Ⅱ (UⅡ) receptor GPR14 in the aorta of apoE knockout mouse. METHODS: The expression of GPR14 in the aorta of apoE knockout C57BL/6J mice at various ages (18 weeks, 28 weeks, and 38 weeks old, respectively) was determined with competitive RT-PCR. A binding assay of [ 125 I]-UⅡ on the aortic tissue was also performed in 28 weeks group. RESULTS: We found significant upregulation of GPR14 mRNA at all three ages. Compared with wild type group at the same age, the GPR14 mRNA level in apoE knockout mice increased 54.2% in 18 week group (P
8.Expression of Hsp27 and Hsp70 in lymphocytes and plasma in healthy workers and coal miners with lung cancer.
Haijiao, WANG ; Jingcai, XING ; Feng, WANG ; Wenhui, HAN ; Houmao, REN ; Tangchun, WU ; Weihong, CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(4):415-20
In coal mines, main occupational hazard is coal-mine dust, which can cause health problem including coal workers' pneumoconiosis and lung cancer. Some heat shock proteins (Hsps) have been reported as an acute response to a wide variety of stressful stimuli. Whether Hsps protect against chronic environmental coal-mine dust over years is unknown. It is also interesting to know that whether the expression of Hsp27 and Hsp70 proteins as a marker for exposure is associated risk of lung cancer among coal miners. We investigated the association between levels of Hsp27 and Hsp70 expression in lymphocytes and plasma and levels of coal-mine dust exposure in workplace or risk of lung cancer in 42 cancer-free non-coal miners, 99 cancer-free coal miners and 51 coal miners with lung cancer in Taiyuan city in China. The results showed that plasma Hsp27 levels were increased in coal miners compared to non-coal miners (P<0.01). Except high cumulative coal-mine dust exposure (OR=13.62, 95%CI=6.05-30.69) and amount of smoking higher than 24 pack-year (OR=2.72, 95% CI=1.37-5.42), the elevated levels of plasma Hsp70 (OR=13.00, 95% CI=5.14-32.91) and plasma Hsp27 (OR=2.97, 95% CI=1.40-6.32) and decreased expression of Hsp70 in lymphocytes (OR=2.36, 95% CI=1.05-5.31) were associated with increased risk of lung cancer. These findings suggest that plasma Hsp27 may be a potential marker for coal-mine dust exposure. And the expression of Hsp27 and Hsp70 levels in plasma and lymphocytes may be used as biomarkers for lung cancer induced by occupational coal-mine dust exposure.
9.Effects of four dental alloys on apoptosis related gene and protein expression of fibroblast L929.
He MENG ; Jie DING ; Ren LI ; Ruiying LIANG ; Wenhui WU
West China Journal of Stomatology 2013;31(3):242-246
OBJECTIVETo investigate the effects of the leaching liquids of 4 differents kinds of dental alloys (Au alloy, Ag-Pd alloy, Co-Cr alloy, Ni-Cr alloy) on apoptosis related gene and protein of fibroblast L929.
METHODSThe L929 cells of mouse were treated in vitro with leaching liquids of 4 different kinds of dental alloys, Au alloy (group A), Ag-Pd alloy(group B), Co-Cr alloy(group C) and Ni-Cr alloy(group D). The RPMI1640 cell medium containing 10% fetal calf serum was served as a control(group E). The effects of these alloys on the expression of caspase-3, 8, 9 of L929 cells were examined by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry method. Results After 48 h culture, the mRNA levels of caspase-3 and caspase-9 demonstrated significant differences between the groups expect group A and group E. The mRNA levels of caspase-8 had no change in all groups. The expression of caspase-3 and caspase-9 were significant differences between the groups expect group A and C, group B and D. The expression of caspase-8 had no change in all grotps.
CONCLUSIONThe leaching liquids of 4 different kinds of dental alloys expect Au alloy may induce cell appotosis through mitochondrion pathway.
Animals ; Apoptosis ; Chromium Alloys ; Dental Alloys ; Fibroblasts ; Materials Testing ; Mice
10.Isolation and biological properties of avian infectious bronchitis virus isolated from Shanxi province
Fang YAN ; Wenbin YUE ; Juan LIU ; Xuying LI ; Yujun ZHAO ; Wenhui JI ; Fengbo LIU ; Qian WU ; Jiayan REN ; Lizhen HUA
Chinese Journal of Veterinary Science 2009;29(7):845-848
Five field strains of infectious bronchitis virus (IBV) were isolated from suspected flocks from different time and different regions of Shanxi province,respectively,and characterized by a series of systematic identification assays,such as morphological observation by electron-microscope,interfering with the propagation of NDV,virus pathological role to chicken embryo,virus pathological role to SPF chickens,hemagglutination activity,physiscochemical,and reverse transcription polymerase chain reaction(RT-PCR).The results showed:The typical coronavirus which the spherical virions 60-120 nm in diameter and surface covered with spike like corona were observed under electron-microscope)The propagation of NDV strain was seriously interfered by the 5 isolates respectively;The embryonated chicken egg passages of the 5 isolates could dwarf with chicken embryos;The five isolates had no hemagglutination activity,but after treatment with 1% trypsin,it can agglutinate chicken red blood cell.The strains are sensitive to chloroform and ethyl ether.The SPF chickens which inoculated with the 5 isolates showed clinical sign and result in respiratory and kidney diseases,flower-steak,and swollen with severe urate deposition.The specific fragments of N gene of the 5 isolates were amplified by RT-PCR,respectiveiy.On the basis of all above mentioned results,the 5 isolates were classified as IBV.The study got a good preparedness for further study on molecular epidemioogy of the 5 IBV isolates.