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AAV2-PDE6B restores retinal structure and function in the retinal degeneration 10 mouse model of retinitis pigmentosa by promoting phototransduction and inhibiting apoptosis
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作者 Ruiqi Qiu Mingzhu Yang +5 位作者 Xiuxiu Jin Jingyang Liu Weiping Wang Xiaoli Zhang Jinfeng Han Bo Lei 《Neural Regeneration Research》 SCIE CAS 2025年第8期2408-2419,共12页
Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-asso... Retinitis pigmentosa is a group of inherited diseases that lead to retinal degeneration and photoreceptor cell death.However,there is no effective treatment for retinitis pigmentosa caused by PDE6B mutation.Adeno-associated virus(AAV)-mediated gene therapy is a promising strategy for treating retinitis pigmentosa.The aim of this study was to explore the molecular mechanisms by which AAV2-PDE6B rescues retinal function.To do this,we injected retinal degeneration 10(rd10)mice subretinally with AAV2-PDE6B and assessed the therapeutic effects on retinal function and structure using dark-and light-adapted electroretinogram,optical coherence tomography,and immunofluorescence.Data-independent acquisition-mass spectrometry-based proteomic analysis was conducted to investigate protein expression levels and pathway enrichment,and the results from this analysis were verified by real-time polymerase chain reaction and western blotting.AAV2-PDE6B injection significantly upregulated PDE6βexpression,preserved electroretinogram responses,and preserved outer nuclear layer thickness in rd10 mice.Differentially expressed proteins between wild-type and rd10 mice were closely related to visual perception,and treating rd10 mice with AAV2-PDE6B restored differentially expressed protein expression to levels similar to those seen in wild-type mice.Kyoto Encyclopedia of Genes and Genome analysis showed that the differentially expressed proteins whose expression was most significantly altered by AAV2-PDE6B injection were enriched in phototransduction pathways.Furthermore,the phototransductionrelated proteins Pde6α,Rom1,Rho,Aldh1a1,and Rbp1 exhibited opposite expression patterns in rd10 mice with or without AAV2-PDE6B treatment.Finally,Bax/Bcl-2,p-ERK/ERK,and p-c-Fos/c-Fos expression levels decreased in rd10 mice following AAV2-PDE6B treatment.Our data suggest that AAV2-PDE6B-mediated gene therapy promotes phototransduction and inhibits apoptosis by inhibiting the ERK signaling pathway and upregulating Bcl-2/Bax expression in retinitis pigmentosa. 展开更多
关键词 apoptosis AAV2-PDE6B ERK1/2 gene therapy PHOTOTRANSDUCTION PROTEOMICS rd10 retinitis pigmentosa
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Neuronal nitric oxide synthase/reactive oxygen species pathway is involved in apoptosis and pyroptosis in epilepsy 被引量:2
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作者 Xiao-Xue Xu Rui-Xue Shi +11 位作者 Yu Fu Jia-Lu Wang Xin Tong Shi-Qi Zhang Na Wang Mei-Xuan Li Yu Tong Wei Wang Miao He Bing-Yang Liu Gui-Lan Chen Feng Guo 《Neural Regeneration Research》 SCIE CAS CSCD 2023年第6期1277-1285,共9页
Dysfunction of neuronal nitric oxide synthase contributes to neurotoxicity,which triggers cell death in various neuropathological diseases,including epilepsy.Studies have shown that inhibition of neuronal nitric oxide... Dysfunction of neuronal nitric oxide synthase contributes to neurotoxicity,which triggers cell death in various neuropathological diseases,including epilepsy.Studies have shown that inhibition of neuronal nitric oxide synthase activity increases the epilepsy threshold,that is,has an anticonvulsant effect.However,the exact role and potential mechanism of neuronal nitric oxide synthase in seizures are still unclear.In this study,we performed RNA sequencing,functional enrichment analysis,and weighted gene coexpression network analysis of the hippocampus of tremor rats,a rat model of genetic epilepsy.We found damaged hippocampal mitochondria and abnormal succinate dehydrogenase level and Na+-K+-ATPase activity.In addition,we used a pilocarpine-induced N2a cell model to mimic epileptic injury.After application of neuronal nitric oxide synthase inhibitor 7-nitroindazole,changes in malondialdehyde,lactate dehydrogenase and superoxide dismutase,which are associated with oxidative stress,were reversed,and the increase in reactive oxygen species level was reversed by 7-nitroindazole or reactive oxygen species inhibitor N-acetylcysteine.Application of 7-nitroindazole or N-acetylcysteine downregulated the expression of caspase-3 and cytochrome c and reversed the apoptosis of epileptic cells.Furthermore,7-nitroindazole or N-acetylcysteine downregulated the abnormally high expression of NLRP3,gasdermin-D,interleukin-1βand interleukin-18.This indicated that 7-nitroindazole and N-acetylcysteine each reversed epileptic cell death.Taken together,our findings suggest that the neuronal nitric oxide synthase/reactive oxygen species pathway is involved in pyroptosis of epileptic cells,and inhibiting neuronal nitric oxide synthase activity or its induced oxidative stress may play a neuroprotective role in epilepsy. 展开更多
关键词 apoptosis bioinformatics analysis cell death EPILEPSY nitric oxide synthase oxidative stress PYROPTOSIS RNA sequencing Tremor rat weighted gene co-expression network analysis
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Analysis of in vivo patterns of caspase 3 gene expression in primary hepatocellular carcinoma and its relationship to p21^(WAF1) expression and hepatic apoptosis 被引量:65
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作者 Bao Hua Sun Jun Zhang Bao JǜWang Xi Ping Zhao You Kun Wang Zhi Qun Yu Dong Liang Yang Lian Jie Hao Department of Clinical Immunology,Tongji Hospital,Tongji Medical University,Wuhan 430030,Hubei Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2000年第3期356-360,共5页
AIM To detect the expression of caspase 3gene in primary human hepatocellular carcinoma(HCC)and investigate its relationship to p21<sup>WAF1</sup>gene expression and HCC apoptosis.METHODS In situ hybridi... AIM To detect the expression of caspase 3gene in primary human hepatocellular carcinoma(HCC)and investigate its relationship to p21<sup>WAF1</sup>gene expression and HCC apoptosis.METHODS In situ hybridization was employedto determine caspase 3 and p21<sup>WAF1</sup>expression inHCC.In situ end-labeling was used to detecthepatocytic apoptosis in HCC.RESULTS Twenty-one of 39(53.8%)cases ofHCC were found to express caspase 3transcripts,while 45.2% of HCC failed toexpress caspase 3.Non-cancerous adjacent livertissues showed more positive caspase 3(87.5%,7/8)as compared with HCC(P【0.05).The expression of caspase 3 is correlated withHCC differentiation,72.2%(13/18)ofmoderately to highly differentiated HCC showedcaspase 3 transcripts positive,while only 38.1%of poorly differentiated HCC harbored caspase 3transcripts(P【0.05).No relationship wasfound between caspase 3 expression and tumorsize or grade or metastasis,although 52.5%(5/8)of HCC with metastasis were caspase 3positive and a little higher than that with nometastasis(51.6%,P】0.05).Expression of caspase 3 alone did not affect the apoptosisindex(AI)of HCC.The AI was 7.12%o in caspase3-positive tumors(n=21),while in caspase 3-negative cases(n=18)6.59%0(P】0.05).Expression of caspase 3 clearly segregated withp21<sup>WAF1</sup>positive tumors as compared withp21<sup>WAF1</sup>-negative cases(16 of 23,69.6% versus5 of 16,31.3%)with statistical significance(P=0.017).In the cases with positive caspase 3and negative p21<sup>WAF1</sup>,the Al was found slightlyhigher,but with no statistical significance,thanthat with expression of p21<sup>WAF1</sup>and caspase 3(7.21‰ vs 6.98‰,P】0.05).CONCLUSION Loss of caspase 3 expressionmay contribute to HCC carcinogenesis,althoughthe expression of caspase 3 does not correlatewell with cell apoptosis in HCC.p21<sup>WAF1</sup>may bemerely one of the inhibitors which can reducecaspase 3 mediated cell apoptosis in HCCs. 展开更多
关键词 carcinoma HEPATOCELLULAR CASPASE 3 apoptosis liver NEOPLASMS gene EXPRESSION
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Apoptosis,proliferation and p53 gene expression of H.pylori associated gastric epithelial lesions 被引量:46
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作者 Zhong Zhang~1 Yuan Yuan Hua Gao Ming Dong Lan Wang Yue-Hua Gong 1 Department of Pathology,Shenyang Medical College,Shenyang 110031 Liaoning Province,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2001年第6期779-782,共4页
AIM: To study the relationship between Helicobacter pylori (H. pylori) and gastric carcinoma and its possible pathogenesis by H. pylori. METHODS: DNEL technique and immunohistochemical technique were used to study the... AIM: To study the relationship between Helicobacter pylori (H. pylori) and gastric carcinoma and its possible pathogenesis by H. pylori. METHODS: DNEL technique and immunohistochemical technique were used to study the state of apoptosis, proliferation and p53 gene expression. A total of 100 gastric mucosal biopsy specimens, including 20 normal mucosa, 30 H. pylori-negative and 30 H. pylori-positive gastric precancerous lesions along with 20 gastric carcinomas were studied. RESULTS: There were several apoptotic cells in the superficial epithelium and a few proliferative cells within the neck of gastric glands, and no p53 protein expression in normal mucosa. In gastric carcinoma, there were few apoptotic cells, while there were a large number of proliferative cells, and expression of p53 protein significantly was increased. In the phase of metaplasia, the apoptotic index (AI, 4.36%+/-1.95%), proliferative index (PI, 19.11%+/-6.79%) and positivity of p53 expression (46.7%) in H. pylori-positive group were higher than those in normal mucosa (P【0.01). AI in H. pylori-positive group was higher than that in H. pylori-negative group (3.81%+/-1.76%), PI in H. pylori-positive group was higher than that in H. pylori-negative group (12.25%+/-5.63%, P【0.01). In the phase of dysplasia, AI (2.31%+/-1.10%) in H. pylori-positive group was lower (3.05%+/-1.29%) than that in H. pylori-negative group, but PI (33.89%+/-11.65%) was significantly higher (22.09+/-8018%, P【0.01). In phases of metaplasia, dysplasia and gastric cancer in the H. pylori-positive group, AIs had an evidently graduall decreasing trend (P【0.01), while PIs had an evidently gradual increasing trend (P【0.05 or P【0.01), and there was also a trend of gradual increase in the expression of p53 gene. CONCLUSION: In the course of the formation of gastric carcinoma, proliferation of gastric mucosa can be greatly increased by H. pylori, and H. pylori can induce apoptosis in the phase of metaplasia, but in the phase of dysplasia H. pylori can inhibit cellular apoptosis. And H. pylori infection can strengthen the expression of mutated p53 gene. 展开更多
关键词 apoptosis gene Expression Helicobacter pylori Cell Division Gastric Mucosa genes p53 Helicobacter Infections Humans Research Support Non-U.S. Gov't Stomach Diseases
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Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma 被引量:27
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作者 Jia-DingMao PeiWu +3 位作者 Xiang-HouXia Ji-QunHu Wen-BinHuang Guo-QiangXu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第5期721-725,共5页
AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma.METHODS: Sixty-two large intestine cancer tissue samples... AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma.METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method.According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed.RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2ss = 9.246; P<0.05,x2GAS = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17)expression groups was higher than that in low expression group (40.0%, 14/35) (P<0.05, x2high vs low = 5.242; P<0.05,x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P<0.05,x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P<0.05, x2ss = 7.178; P<0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11)and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36)(P<0.05,x2high vs low = 5.600; P<0.05, x2 middle vs low = 7.695).However, the positive expression rate of bcl-2 in SS high (40.0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35)(P<0.05, x2 high vs low = 4.710; P<0.05, x2 middle vs low = 4.706).There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P<0.01,r=0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P<0.05, r = -0.299).CONCLUSION: The regulation and control of gastrin,somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax. 展开更多
关键词 Large intestine carcinoma GASTRIN SOMATOSTATIN bcl-2 gene Bax gene apoptosis
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Dynamic expression of apoptosis-related genes during development of laboratory hepatocellular carcinoma and its relation to apoptosis 被引量:38
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作者 Xiao-Xian Duan Jing-Sheng Ou Yuan Li Jian-Jia Su Chao Ou Chun Yang Hui-Fen Yue Ke-Chen Ban 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第30期4740-4744,共5页
AIM: To explore the expression of p53, bcl-2, bax, survivin and the cell apoptosis during the development of tree shrew hepatocellular carcinoma (HCC), the relationship between expression of these genes, its impact... AIM: To explore the expression of p53, bcl-2, bax, survivin and the cell apoptosis during the development of tree shrew hepatocellular carcinoma (HCC), the relationship between expression of these genes, its impact on HCC development, and its relation to cell apoptosis. METHODS: Tree shrew HCC was induced with aflatoxin B1 (AFB1), and regular biopsy of liver tissues was carried out and the biopsy tissues were collected during cancer inducement. Liver biopsy tissue and HCC tissue were collected from 35 pre-cancerous experimental animals at wk 30 and 60 and at the 30^th, 60^th, and 90^th -wk. Liver biopsy tissues were collected from 13 blank control animals at wk 30, 60, and 90. Expression of p53, bcl-2, bax, and survivin at each stage was examined by immunohistochemistry method. Apoptotic cells were detected in situ by the terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) technique. RESULTS: The apoptosis rate of normal hepatic cells was extremely low, whereas it increased during the formation of HCC. Expression of the apoptosis-related genes p53, bcl-2, bax, and survivin during the formation of HCC presented an increasing tendency. Expression of p53 did not noticeably relate to that of bcl-2, bax, and survivin, whereas expression of bcl-2 and bax was closely related. In HCC, p53 did not present a distinct relation to cell apoptosis, whereas its high level expression was probably related to liver cell proliferation. Survivin negatively correlated apoptosis index, and its overexpression could inhibit cell apoptosis. CONCLUSION: Apoptosis-related genes p53, bcl-2, bax, and survivin are all related to the occurrence of HCC. The anti-apoptosis effect of bcl-2 is influenced by bax, and ratio bcl/bax reflects more correctly the extent of cell apoptosis. 展开更多
关键词 Hepatocellular carcinoma apoptosis gene
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Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721 被引量:42
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作者 Sheng-QuanCheng Wen-LiangWang +3 位作者 WeiYan Qing-LongLi LiWang Wen-YongWang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第5期756-759,共4页
AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 c... AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SW), were constructed by ligating into the vector, pSupperssorNeo (pSuNeo) digested with restriction enzymes Xba I and Sail and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidin-biotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi. RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SW, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent, apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited. CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells. 展开更多
关键词 Hepatocellular carcinoma Survivin RNA interference apoptosis gene expression
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miR-29a up-regulation in AR42J cells contributes to apoptosis via targeting TNFRSF1A gene 被引量:7
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作者 Qiang Fu Tao Qin +6 位作者 Lin Chen Chuan-Jiang Liu Xu Zhang Yu-Zhu Wang Ming-Xing Hu Hao-Yuan Chu Hong-Wei Zhang 《World Journal of Gastroenterology》 SCIE CAS 2016年第20期4881-4890,共10页
AIM: To investigate the expression of mi R-29 a in rat acute pancreatitis and its functional role in AR42 J cell apoptosis.METHODS: Twelve SD rats were divided into a control group and an acute edematous pancreatitis(... AIM: To investigate the expression of mi R-29 a in rat acute pancreatitis and its functional role in AR42 J cell apoptosis.METHODS: Twelve SD rats were divided into a control group and an acute edematous pancreatitis(AEP) group randomly. AEP was induced by intraperitoneal injection of L-arginine(150 mg/kg) in the AEP group and equal volume of 0.9% Na Cl was injected in the control group. The apoptosis of acinar cells in pancreatic tissue was determined by TUNEL assay. mi RNA chip assay was performed to examine the expression of mi RNAs in two groups. Besides, to further explore the role of mi R-29 a in apoptosis in vitro, recombinant rat TNF-α(50 ng/m L) was administered to treat the rat pancreatic acinar cell line AR42 J for inducing AR42 J cell apoptosis. Quantitative real-time PCR(q RT-PCR) was adopted to measure mi R-29 a expression. Then, mi RNA mimic, mi RNA antisense oligonucleotide(AMO) and control vector were used to transfect AR42 J cells. The expression of mi R-29 a was confirmed by q RT-PCR andthe apoptosis rate of AR42 J cells was detected by flow cytometry analysis. Western blot was used to detect the expression of activated caspase3. Moreover, we used bioinformatics software and luciferase assay to test whether TNFRSF1 A was the target gene of mi R-29 a. After transfection, q RT-PCR and Western blot was used to detect the expression of TNFRSF1 A in AR42 J cells after transfection.RESULTS: The expression of mi R-29 a was much higher in the AEP group compared with the control group as displayed by the mi RNA chip assay. After inducing apoptosis of AR42 J cells in vitro, the expression of mi R-29 a was significantly increased by 1.49 ± 0.04 times in comparison with the control group. As revealed by q RT-PCR assay, the expression of mi R-29 a was 2.68 ± 0.56 times higher in the mi R-29 a mimic group relative to the control vector group, accompanied with an obviously increased acinar cell apoptosis rate(42.83 ± 1.25 vs 24.97 ± 0.15, P < 0.05). Moreover, the expression of mi R-29 a in the mi RNA AMO group was 0.46 ± 0.05 times lower than the control vector group, and the cell apoptosis rate was much lower accordingly(17.27 ± 1.36 vs 24.97 ± 0.15, P < 0.05). The results of bioinformatics software and luciferase assay showed that TNFRSF1 A might be a target gene of mi R-29 a. TNFRSF1 A expression was up-regulated in the mi R-29 a mimic group, while the mi R-29 a AMO group showed the reverse trend.CONCLUSION: mi R-29 a might promote the apoptosis of AR42 J cells via up-regulating the expression of its target gene TNFRSF1 A. 展开更多
关键词 Acute EDEMATOUS PANCREATITIS mi R-29a apoptosis AR42J Target gene TNFRSF1A
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Effect of TSLC1 Gene on Proliferation, Invasion and Apoptosis of Human Hepatocellular Carcinoma Cell Line HepG2 被引量:8
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作者 覃莉 祝文涛 +4 位作者 许涛 郝友华 张正茂 田拥军 杨东亮 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2007年第5期535-537,共3页
The recombinant plasmid pCI-TSLC1 carrying TSLC1 gene was stably transfected into human hepatocellular carcinoma cell line HepG2. Cell proliferation was analyzed by MTT assay. The ability of migration was determined b... The recombinant plasmid pCI-TSLC1 carrying TSLC1 gene was stably transfected into human hepatocellular carcinoma cell line HepG2. Cell proliferation was analyzed by MTT assay. The ability of migration was determined by transwell and FACSort flow cytometry was used to detect the cell cycle distribution and apoptosis. Western blotting revealed that H4 expressed higher amounts of TSLC1 protein than H15 and H0 did. The growth of TSLCl-transfected cells was significantly suppressed in vitro, and the ability of migration was reduced as well. The re-expression of TSLC1 could induce cell apoptosis. It was concluded that TSLC1 strongly inhibited the growth and ability of migration of HepG2 cell line in vitro and also induced apoptosis, suggesting that TSLC 1 could reduce the tumorigenicity of human hepatocellular carcinoma cell line HepG2 in vitro, which provided a basis for further exploring the roles of TSLC1 in hepatocellular cellular carcinoma. 展开更多
关键词 carcinoma hepatocellular TSLC1 gene PROLIFERATION INVASION apoptosis
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^(103)Pd-induced apoptosis of proliferative smooth muscle cells in bile ducts of dogs:significance and effects on related genes 被引量:5
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作者 Gui-Jin He,Fa-Qiang Yu,Rong Wu,Qin-Yi Gao,Shu-He Xu,Hong Gao, Wei-Guo Jiang,Tao Jiang and Xian-Wei Dai Department of General Surgery,Second Affiliated Hospital(Shengjing Hospital),China Medical University,Shenyang 110004,China,Radiotherapy Center,Second Affiliated Hospital(Shengjing Hospital),China Medical University,Shenyang 110004,China,Pediatrics Surgery Laboratory,Second Affiliated Hospital(Shengjing Hospital),China Medical University,Shenyang 110004,China,Department of Pathology,Second Affiliated Hospital(Shengjing Hospital),China Medical University,Shenyang 110004,China Isotope Unit,First Affiliated Hospital,China Medical University,Shenyang 110001,China and Isotope Research Institute,China Institute of Atomic Energy,Beijing 100050,China 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2007年第5期521-526,共6页
BACKGROUND:With the objective of developing a locally- produced radioactive stent,the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation... BACKGROUND:With the objective of developing a locally- produced radioactive stent,the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation of the expression of genes caused byγ-radiation in order to prevent bile duct restenosis.We therefore explored the effects and significance ofγ-radiation on the activity of caspase-3,Fas and Bcl-2 genes in apoptosis of proliferative smooth muscle cells in the bile duct walls of dogs. METHODS:Twelve dogs were randomly divided into 2 groups(6 in each group).A postinjury bile duct stenosis model was established and radioactive 103 Pd( 103 palladium) or ordinary bile duct stents were implanted into the bile ducts.HE staining,RT-PCR and immunohistochemistry were used to detect the proliferation and apoptosis of bile duct smooth muscle cells in proliferative endomembrane and the expression of related caspase-3,Bcl-2 and Fas genes. RESULTS:The expression of caspase-3 and Fas genes in the bile duct tissues of dogs with radioactive stents was higher than that of dogs with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the bile ducts.The expression of the Bcl-2 gene in the bile duct tissues of dogs with radioactive stents was lower than that in those with ordinary stents.There was significant apoptosis of proliferative smooth muscle cells in the dogs with low Bcl-2 gene expression. CONCLUSIONS:Radiation increases the activity of caspase-3 and Fas genes and is associated with apoptosis. The radioactive 103 Pd stent may facilitate apoptosis of proliferative smooth muscle cells in the bile ducts of dogs by activating these genes.The Bcl-2 gene expression level is correlated with the occurrence of apoptosis and the radiosusceptibility of cells. 展开更多
关键词 radioactive 103 Pd stent γ-radiation caspase-3 gene Fas gene Bcl-2 gene apoptosis
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Protein Expression of BLM Gene and Its Apoptosis Sensitivity in Hematopoietic Tumor Cell Strains 被引量:6
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作者 王晓蓓 胡丽华 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2008年第1期46-48,共3页
Patients with Bloom syndrome (BS) show an immunodeficiency, an enhanced sister chromatid exchanges (SCEs), a strong genetic instability and an increased predisposition to all. In order to investigate the different... Patients with Bloom syndrome (BS) show an immunodeficiency, an enhanced sister chromatid exchanges (SCEs), a strong genetic instability and an increased predisposition to all. In order to investigate the differential expression of BLM protein in hematopoietic tumor cell strains and study the effects of BLM gene on ultraviolet (UV)- or hydroxyurea (HU)-induced apoptosis, Western blot was used to detect the expression of BLM protein in normal human bone marrow mononuclear cells and 4 kinds of hematopoietic tumor cell strains. The 4 kinds of hematopoietic tumor cells were exposed to UV light with a germicidal UV lamp or treated with 2 mmol/L hydroxyurea and the apoptotic rate was detected by using AnnexinV-FITC. The results showed that these tumor cells expressed BLM protein higher than the normal human bone marrow mononuclear cells (P〈0.01). In the 4 hematopoietic tumor cells, BLM protein was all specially cleaved in response to UV- or HU-induced apoptosis. The increase of BLM protein expression may play an important role in the development of these tumors, and BLM proteolysis is likely to be a general feature of the apoptotic response. 展开更多
关键词 BLM gene PROTEIN apoptosis hematopoietic tumor cell strains
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Targeted IL-24 gene therapy inhibits cancer recurrence after liver tumor resection by inducing tumor cell apoptosis in nude mice 被引量:5
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作者 Yang, Yong-Jiu Chen, Da-Zhi +3 位作者 Li, Li-Xin Sheng, Qin-Song Jin, Zhong-Kui Zhao, De-Fang 《Hepatobiliary & Pancreatic Diseases International》 SCIE CAS 2009年第2期174-178,共5页
BACKGROUND: Interleukin-24 (IL-24) is a novel candidate tumor suppressor that induces tumor cell apoptosis experimentally in a variety of human malignant cells including liver cancer cells. The present study was condu... BACKGROUND: Interleukin-24 (IL-24) is a novel candidate tumor suppressor that induces tumor cell apoptosis experimentally in a variety of human malignant cells including liver cancer cells. The present study was conducted to investigate the potential effect of recombinant adeno-associated virus (rAAV)-mediated IL-24 gene therapy on tumor recurrence and metastasis by inducing tumor cell apoptosis in a hepatocellular carcinoma (HCC) model in nude mice. METHODS: We established a recurrent and metastatic HCC model in nude mice and constructed an rAAV vector carrying alpha-fetoprotein (AFP) promoter for expressing the IL-24 gene (rAVV/AFP/IL-24). The vector was administered by regional injection (liver incisal margin). AFP was detected by radiation immunoassay. Histological evaluation of tumor recurrence and metastasis was performed for the liver and lung. The effect of tumor cell apoptosis was confirmed by TUNEL analysis. RESULTS: IL-24 gene therapy prevented tumor recurrence and metastasis, as evidenced by marked decreases in the number of metastatic tumor nodules and tumor volume in the liver and lung. At the same time, serum AFP concentration decreased markedly in the IL-24 group compared with the control or rAAV groups (P<0.05). IL-24 gene therapy inhibited tumor recurrence and metastasis as evidenced by the induction of tumor cell apoptosis. CONCLUSION: The results demonstrated that targeted IL-24 gene therapy was effective in the prevention of postoperative recurrence and metastasis in an HCC nude mice model by induction of tumor cells apoptosis with potential minimum tumor burden. 展开更多
关键词 liver neoplasms gene therapy INTERLEUKIN-24 apoptosis
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Possible mechanism for hepatitis B virus X gene to induce apoptosis of hepatocytes 被引量:12
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作者 Sheng-Jun Zhang Hong-Ying Chen +1 位作者 Zhi-Xin Chen Xiao-Zhong Wang 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第28期4351-4356,共6页
AIM: To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by... AIM: To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells.METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 μg/mL G418 and named HL-7702/HBV-encoded X protein (HBx) cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively.RESULTS: RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semiquantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than thosein control.CONCLUSION: HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner. 展开更多
关键词 Hepatitis B virus X gene apoptosis geneEXPRESSION
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Growth arrest-specific gene 2 suppresses hepatocarcinogenesis by intervention of cell cycle and p53-dependent apoptosis 被引量:4
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作者 Ran-Xu Zhu Alfred Sze Lok Cheng +2 位作者 Henry Lik Yuen Chan Dong-Ye Yang Wai-Kay Seto 《World Journal of Gastroenterology》 SCIE CAS 2019年第32期4715-4726,共12页
BACKGROUND Growth arrest-specific gene 2(GAS2)plays a role in modulating in reversible growth arrest cell cycle,apoptosis,and cell survival.GAS2 protein is universally expressed in most normal tissues,particularly in ... BACKGROUND Growth arrest-specific gene 2(GAS2)plays a role in modulating in reversible growth arrest cell cycle,apoptosis,and cell survival.GAS2 protein is universally expressed in most normal tissues,particularly in the liver,but is depleted in some tumor tissues.However,the functional mechanisms of GAS2 in hepatocellular carcinoma(HCC)are not fully defined.AIM To investigate the function and mechanism of GAS2 in HCC.METHODS GAS2 expression in clinic liver and HCC specimens was analyzed by real-time PCR and western blotting.Cell proliferation was analyzed by counting,MTS,and colony formation assays.Cell cycle analysis was performed by flow cytometry.Cell apoptosis was investigated by Annexin V apoptosis assay and western blotting.RESULTS GAS2 protein expression was lower in HCC than in normal tissues.Overexpression of GAS2 inhibited the proliferation of HCC cells with wide-type p53,while knockdown of GAS2 promoted the proliferation of hepatocytes(P<0.05).Furthermore,GAS2 overexpression impeded the G1-to-S cell cycle transition and arrested more G1 cells,particularly the elevation of sub G1(P<0.01).Apoptosis induced by GAS2 was dependent on p53,which was increased by etoposide addition.The expression of p53 and apoptosis markers was further enhanced when GAS2 was upregulated,but became diminished upon downregulation of GAS2.In the clinic specimen,GAS2 was downregulated in more than 60%of HCCs.The average fold changes of GAS2 expression in tumor tissues were significantly lower than those in paired non-tumor tissues(P<0.05).CONCLUSION GAS2 plays a vital role in HCC cell proliferation and apoptosis,possibly by regulating the cell cycle and p53-dependent apoptosis pathway. 展开更多
关键词 Growth arrest-specific gene 2 Cell cycle apoptosis Hepatocellular carcinoma p53-dependent signaling pathway
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Melanoma differentiation-associated gene-7, MDA-7/IL-24, selectively induces growth suppression, apoptosis in human hepatocellular carcinoma cell line HepG2 by replication-incompetent adenovirus vector 被引量:15
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作者 Cong-Jun Wang Xin-Bo Xue Ji-Lin Yi Kun Chen Jian-WeiZheng Jian Wang Jian-Ping Zeng Rong-Hua Xu 《World Journal of Gastroenterology》 SCIE CAS CSCD 2006年第11期1774-1779,共6页
AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma (HCC) cell line HepG2 and normal liver cell line L0... AIM: To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma (HCC) cell line HepG2 and normal liver cell line L02. METHODS: We constructed the recombinant replication-incornpetent Ad.rnda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the rnRNA expressing in cells, by ELISA was used to detect MDA-7/IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.rnda-7 was confirmed by Hoechst staining and flow cytometry assay with Annexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. RESULTS: Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RTPCR showed that the Ad.rnda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.rnda-7-infected L02 and HepG2 ceils, respectively. Ad.mda-7 infection obviously induced apoptosis (from 2.604±0.72% to 33.64±13.2%, P=0.00012) and growth suppression in HepG2 (inhibition ratio IR=68%) and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle (from 6.44% to 32.29%, P〈 0.01), but not in L02 cells.CONCLUSION: These results confirm selectively induction of apoptosis and growth suppression by the mda-7/ IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2. 展开更多
关键词 Cancer gene therapy Hepatocellular carcinoma (HCC) apoptosis Growth suppression MDA-7/IL-24
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Suppressive Effects of Genomic Imprinted Gene PEG10 on Hydrogen Peroxide-induced Apoptosis in L0_2 Cells 被引量:3
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作者 刘瑶 黄焕军 +3 位作者 林菊生 张强 谭锦泉 任精华 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2009年第6期705-709,共5页
The effects of PEG10 on hydrogen peroxide (H2O2)-induced apoptosis in human normal liver cell line L02 were investigated. The PEG10 gene was transfected into L02 cells by lipofectamine, the positive clone was screen... The effects of PEG10 on hydrogen peroxide (H2O2)-induced apoptosis in human normal liver cell line L02 were investigated. The PEG10 gene was transfected into L02 cells by lipofectamine, the positive clone was screened by G418 and defined as L02/PEG10, while the cell transfected with empty expression vector (pEGFP-N1) was defined as L02/vector. L02/vector and parental L02 cells served as control. RT-PCR and Western blotting were employed to detect the expression of target genes. H2O2 (50–400 mmol/L) was administered to induce the apoptosis of L02 cells. Cells viability was measured by MTT and the morphological changes of apoptotic cells were determined by fluorescence microscopy using hoechst33342 nuclei staining. DNA fragmentation was observed by agarose gel electrophoresis. PEG10 mRNA and protein levels in L02/PEG10 cells were significantly increased as compared with those in the control cells. After treatment with 400 mmol/L H2O2 for 24 h, the cellular growth inhibition rate of L02/PEG10 cells was significantly lower (58.2%) than that of L02 (92.5%) and L02/vector (88%). Distinct morphological changes characteristic of cell apoptosis such as karyopyknosis and conglomeration were not observed in L02/PEG10. Ladder-like DNA fragmentation in a dose-dependent manner was observed in both L02 and L02/vector cell lines, but not in L02/PEG10. PEG10 over-expression significantly inhibited cytotoxicity induced by H2O2 on human normal liver cell line L02 by antagonizing H2O2-induced apoptosis. 展开更多
关键词 genetic imprinting gene PEG10 L02 hepatocytes hydrogen peroxide apoptosis
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Stable transfection of extrinsic Smac gene enhances apoptosis-inducing effects of chemotherapeutic drugs on gastric cancer cells 被引量:5
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作者 Li-DuanZheng Qiang-SongTong +2 位作者 LiangWang JunLiu WeiQian 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第1期79-83,共5页
AIM: To explore the feasibility of enhancing apoptosis-inducing effects of chemotherapeutic drugs on human gastric cancer cells by stable transfection of extrinsic Smac gene. METHODS: After Smac gene was transferred i... AIM: To explore the feasibility of enhancing apoptosis-inducing effects of chemotherapeutic drugs on human gastric cancer cells by stable transfection of extrinsic Smac gene. METHODS: After Smac gene was transferred into gastric cancer cell line MKN-45, subclone cells were obtained by persistent G_(418) selection. Cellular Smac gene expression was determined by RT-PCR and Western blotting. After treatment with mitomycin (MMC) as an apoptotic inducer, in vitro cell growth activities were investigated by trypan blue-staining method and MTT colorimetry. Cell apoptosis and its rates were determined by electronic microscopy, annexin V-FTTC and propidium iodide staining flow cytometry. Cellular caspase-3 protein expression and its activities were assayed by Western blotting and colorimetry. RESULTS: When compared with MKN-45 cells, the selected subclone cell line MKN-45/Smac had significantly higher Smac mRNA (3.12±0.21 vs 0.82±0.14, t=7.52, P<0.01) and protein levels (4.02±0.24 vs0.98±0.11, t=8.32, P<0.01). After treatment with 10 μg/mL MMC for 6-24 h, growth inhibition rate of MKN-45/Smac (15.8±1.2-54.8±2.9%) was significantly higher than that of MKN-45 (5.8±0.4-24.0±1.5%, t=6.42, P<0.01). Partial MKN-45/Smac cancer cells presented characteristic morphological changes of apoptosis under the electronic microscope with an apoptosis rate of 36.4±2.1%, which was significantly higher than that of MKN-45 (15.2±0.8%, t=9.25, P<0.01). Compared with MKN-45, caspase-3 expression levels in MKN-45/Smac were improved significantly (3.39±0.42 vs0.96±0.14, t=8.63, P<0.01), while its activities were 3.25 times as many as those of MKN-45 (0.364±0.010 vs0.112±0.007, t=6.34, P<0.01). CONCLUSION: Stable transfection of extrinsic Smac gene and its over-expression in gastric cancer cell line can significantly enhance cellular caspase-3 expression and activities, ameliorate apoptosis-inducing effects of mitomycin C on cancer cells, which is a novel strategy to improve chemotherapeutic effects on gastric cancer. 展开更多
关键词 Gastric cancer Mitomycin C Extrinsic Smac gene apoptosis TRANSFECTION
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Cloning of Chicken Anemia Virus vp3 Gene and Apoptosis Inductive Effect of vp3 Gene In Vitro 被引量:2
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作者 孙军 王宇哲 +1 位作者 宗义强 屈伸 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2003年第4期329-331,334,共4页
Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA vp3. Restriction enzyme digestion and sequencing analysis revea... Using PCR technique, the vp3 gene of chicken anemia virus (CAV) was cloned into the eukaryotic expression vector pcDNA3 to construct a recombinant pcDNA vp3. Restriction enzyme digestion and sequencing analysis revealed that CAV vp3 gene was correctly inserted into the blank vector pcDNA3. After LipofectAMINE TM mediated transfection in vitro with pcDNA vp3 and pcDNA3 respectively, the total mRNA was extracted from liver carcinoma cell lines HepG2 and diploid cell line L 02, and RT PCR was performed afterward. The results of RT PCR suggested that vp3 gene was expressed in these two cell lines. At the same time, using in situ apoptotic detection assay, TUNEL kits, the apoptotic cells were found in pcDNA vp3 transfected HepG2, but not in mock transfected cell lines. VP3 could induce cell death by apoptosis in cancer cell lines, but not in diploid cell lines. All the results indicated that CAV vp3 gene, a potential therapeutic agents, has the potential of being used for cancer treatment. 展开更多
关键词 chicken anemia virus gene cloning apoptosis vp3 gene
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Effects of Carvedilol on Cardiomyocyte Apoptosis and Gene Expression In Vivo after Ischemia-Reperfusion in Rats 被引量:2
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作者 曾和松 刘晓春 赵华月 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2003年第2期127-130,共4页
The effects of carvedilol on cardiomyocyte apoptosis and expression of bcl-2. bax genes following ischemia (0. 5 h) and reperfusion (48 h) in vivo and the possible biological mechanism of carvedilol inhibiting cardiom... The effects of carvedilol on cardiomyocyte apoptosis and expression of bcl-2. bax genes following ischemia (0. 5 h) and reperfusion (48 h) in vivo and the possible biological mechanism of carvedilol inhibiting cardiomyocyte apoptosis were studied. The left anterior descending artery in Wistar rats were ligated to establish ischemia-reperfusion (I/R) models. The model animals were divided into two groups: I/R group, the model rats not subject to other treatments except ischemia-reperfusion (n= 8); carvedilol-treated group (n= 8), I/R model rats treated with carvedilol. Eight rats in the sham-operated group were subjected to only experimental open operation. The number of apoptotic cardiomyocyte was determined by TUNEL staining. Immunohistochemistry and in situ hybridization histochemistry (ISHH) were used to detect the expression of bcl-2 and bax genes. Image processing system was used to quantitatively dispose the positive metric substances of both immuno-histochemistry and ISHH through the average optical density (OD) value. The results showed that the number of the apoptotic cells were 36. 18±9 (I/R group) . 0-1 (sham-operated group) and 9. 5± 3 (carvedilol-treated group) in each visual field respectively with the difference being very significant among the groups (P<0. 001). The OD values of bcl-2 protein in sham-operated group, I/R group and carvedilol-treated group were 0. 14±0. 01, 0. 08±0. 02 and 0. 15±0. 03, respectively. The OD values of bcl-2 mRNA in sham-operated group, I/R group and carvedilol-treated group were 0. 08± 0. 01, 0. 06±0. 01 and 0. 09±0. 01, respectively. There was no significant difference between carvedilol-treated group and I/R group (P>0. 05). The OD values of bax protein in I/R group, sham-operated and carvedilol-treated-treated group were 0. 13±0. 02, 0. 07±0. 01, 0. 06±0. 01, respectively. There was very significant difference between carvedilol-treated group and I/R group (P <0. 01). Bcl-2/bax ratio was 1. 07±0. 14 (I/R group), 1. 28±0. 16 (sham-operated group), 2. 5 ±0. 26 (carvedilol-treated group) respectively with the difference being very significant between carvedilol-treated group and I/R group (P<0. 05). It was indicated that carvedilol could inhibit cardiomyocyte apoptosis following ischemia and reperfusion, which was related to the increased bcl-2/ bax ratio due to inhibition of bax gene expression. 展开更多
关键词 CARVEDILOL ISCHEMIA-REPERFUSION apoptosis gene expression
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Expression Pattern of Testis-specific Expressed Gene 2 in Cryptorchidism Model and Its Role in Apoptosis of Spermatogenic Cells 被引量:2
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作者 胡涛 王智宇 +4 位作者 曾甫清 陈晓春 顾朝辉 郑丽端 童强松 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2010年第2期193-197,共5页
In our previous study, we identified a novel testis-specific expressed gene 2 (TSEG-2) from mouse testis. To further investigate its functions, 35 male Balb/c mice (8 weeks old) were divided into cryptorchidism gr... In our previous study, we identified a novel testis-specific expressed gene 2 (TSEG-2) from mouse testis. To further investigate its functions, 35 male Balb/c mice (8 weeks old) were divided into cryptorchidism group (n=20), sham group (n=10), and control group (n=5). In cryptorchidism group, the right testes were anchored to the inner lateral abdominal wall. In situ hybridization (ISH) was applied to measure the localization of TSEG-2 in mouse testis. Real-time quantitative PCR was performed to detect the expression of TSEG-2 gene. Meanwhile, under the mediation of polyethylenimine (PEI), the recombinant vector pEGFP-TSEG-2 (n=5) or empty vector (mock, n=5) was transfected into the testis of male mice. The transfection efficiencies were measured under a fluorescence microscope. The apoptosis of spermatogenic cells was detected by terminal deoxynuleotidyl-mediated nick end labeling (TUNEL). The results showed that TSEG-2 was expressed in convoluted seminiferous tubules, more precisely, in spermatogonia and spermatocytes. As compared with sham and control groups, the TSEG-2 transcription was significantly enhanced (P〈0.05) and was correlated with apoptosis of spermatogenic cells in cryptorchid testes (P〈0.05). PEI was efficient in mediating transfeetion of TSEG-2 into seminiferous tubules of testis. One week post-transfection, intratesticular injection of TSEG-2 resulted in increased apoptosis of spermatogenic cells in vivo (P〈0.05). These results indicate that TSEG-2 may participate in the apoptosis of spermatogenic cells and the pathogenesis of cryptorchidism. 展开更多
关键词 animal model testis-specific expressed gene 2 polyethylenimine apoptosis
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