Pub Date : 2022-01-01Epub Date: 2022-11-22DOI: 10.3389/fstro.2022.1010928
Elizabeth S Fisher, Yanan Chen, Mikaela M Sifuentes, Jeremy J Stubblefield, Damian Lozano, Deborah M Holstein, JingMei Ren, Matthew Davenport, Nicholas DeRosa, Tsung-Pei Chen, Gerard Nickel, Theodore E Liston, James D Lechleiter
Acute ischemic stroke (AIS) is the second leading cause of death globally. No Food and Drug Administration (FDA) approved therapies exist that target cerebroprotection following stroke. Our group recently reported significant cerebroprotection with the adenosine A1/A3 receptor agonist, AST-004, in a transient stroke model in non-human primates (NHP) and in a preclinical mouse model of traumatic brain injury (TBI). However, the specific receptor pathway activated was only inferred based on in vitro binding studies. The current study investigated the underlying mechanism of AST-004 cerebroprotection in two independent models of AIS: permanent photothrombotic stroke in mice and transient middle cerebral artery occlusion (MCAO) in rats. AST-004 treatments across a range of doses were cerebroprotective and efficacy could be blocked by A3R antagonism, indicating a mechanism of action that does not require A1R agonism. The high affinity A3R agonist MRS5698 was also cerebroprotective following stroke, but not the A3R agonist Cl-IB-MECA under our experimental conditions. AST-004 efficacy was blocked by the astrocyte specific mitochondrial toxin fluoroacetate, confirming an underlying mechanism of cerebroprotection that was dependent on astrocyte mitochondrial metabolism. An increase in A3R mRNA levels following stroke suggested an intrinsic cerebroprotective response that was mediated by A3R signaling. Together, these studies confirm that certain A3R agonists, such as AST-004, may be exciting new therapeutic avenues to develop for AIS.
{"title":"Adenosine A1R/A3R agonist AST-004 reduces brain infarction in mouse and rat models of acute ischemic stroke.","authors":"Elizabeth S Fisher, Yanan Chen, Mikaela M Sifuentes, Jeremy J Stubblefield, Damian Lozano, Deborah M Holstein, JingMei Ren, Matthew Davenport, Nicholas DeRosa, Tsung-Pei Chen, Gerard Nickel, Theodore E Liston, James D Lechleiter","doi":"10.3389/fstro.2022.1010928","DOIUrl":"10.3389/fstro.2022.1010928","url":null,"abstract":"<p><p>Acute ischemic stroke (AIS) is the second leading cause of death globally. No Food and Drug Administration (FDA) approved therapies exist that target cerebroprotection following stroke. Our group recently reported significant cerebroprotection with the adenosine A1/A3 receptor agonist, AST-004, in a transient stroke model in non-human primates (NHP) and in a preclinical mouse model of traumatic brain injury (TBI). However, the specific receptor pathway activated was only inferred based on <i>in vitro</i> binding studies. The current study investigated the underlying mechanism of AST-004 cerebroprotection in two independent models of AIS: permanent photothrombotic stroke in mice and transient middle cerebral artery occlusion (MCAO) in rats. AST-004 treatments across a range of doses were cerebroprotective and efficacy could be blocked by A3R antagonism, indicating a mechanism of action that does not require A1R agonism. The high affinity A3R agonist MRS5698 was also cerebroprotective following stroke, but not the A3R agonist Cl-IB-MECA under our experimental conditions. AST-004 efficacy was blocked by the astrocyte specific mitochondrial toxin fluoroacetate, confirming an underlying mechanism of cerebroprotection that was dependent on astrocyte mitochondrial metabolism. An increase in A3R mRNA levels following stroke suggested an intrinsic cerebroprotective response that was mediated by A3R signaling. Together, these studies confirm that certain A3R agonists, such as AST-004, may be exciting new therapeutic avenues to develop for AIS.</p>","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"36 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10861240/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88398541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.010
Xuzhi Zhang, Ao Ren, Zhongqiu Li, R. Deng, Yi Ma
Objective To study the effect of inhibiting NLRC5 on CD4+ T cells function and allografts immune protective effects in mice. Methods Mice spleen-derived CD4+ T cells were cultured, purified andtransfected with short hairpin RNA (shRNA) lentiviral vector NLRC5-RNA interference (RNAi)-green fluorescent protein (GFP). The mice allogenic islet transplantation model andskintransplantation model were established, and 24 h prior to transplantation each recipient mice was given CD4+ T cells, NLRC5-RNAi-T cells or equal amount of phosphate buffer (PBS), named T cell group, NLRC5-RNAi group and control group respectively. The survival of islet grafts andskingraftsof transplanted recipients was observed, and on the day 7, the islet graft glucose tolerance, the percentage of T cell subsets in the spleenaswellas related cytokines interleukin (IL)-10 and interferon (IFN)-γwere observed. Results The experimental data demonstratedthat NLRC5-RNAi significantly prolonged the survival timewith a median survival time of (19.0±3.4) days inislet allograftand (15.4±3.8) days inskin allograft, which is significantly longer than control [(11.6±1.9) d, t=5.156, P<0.01] and T cell group [(10.0±1.4) d, t=4.151, P<0.01] inislet allograft, aswellascontrol [(8.0±0.9) d, t=3.375, P<0.05] and T cell group [(7.8±0.8) d, t=3.375, P<0.05] inskin allograft. The results of enzyme linked immunosorbent assay (ELISA) showed that the expression of IL-10 (344.0±4.1) ng/L inisletallograft and (275.9±12.5) ng/L in skinallograftin NLRC5-RNAi group were significantly higher than control group (t=124.141, 20.121, P<0.01) and T cell group (t=32.605, 17.900, P<0.01); the expression of IFN-γ (85.1±6.6) ng/L inisletallograftand (96.6±2.5) ng/L inskinallograftin NLRC5-RNAi group were lower than control group (t=7.633, 7.490, P<0.05) and T cell group (t=10.972, 13.286, P<0.01). Flow cytometric analysis revealed that compared to control group and T cell group, the NLRC5-RNAi group dramatically increased the population of Th2 [(0.190±0.053)%, t=5.220, 5.278, P<0.05] inislet allograft and [(0.130±0.012)%, t=21.060, 9.470, P<0.05] in skin allograft, as well asreduced the population of Th1 [(0.810±0.036)%, t=6.219, 5.276, P<0.05] inislet allograft and [(0.180±0.026)%, t=9.248, 25.324, P<0.05] in skin allograft. Conclusion After inhibiting the expression of NLRC5 gene, the expression of Th2 type cytokines in CD4+ T cells is increased, which prolongs the survival time of grafts and has positive significance for the induction of transplantation tolerance. Key words: NLRC5; Lentivirus transduction; CD4+ T cells; Islet transplantation; Skin transplantation; Immune tolerance
{"title":"Inhibit NLRC5 regulate CD4+ T cellsfunctionandimmunological protection onallograftinmice","authors":"Xuzhi Zhang, Ao Ren, Zhongqiu Li, R. Deng, Yi Ma","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.010","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.010","url":null,"abstract":"Objective \u0000To study the effect of inhibiting NLRC5 on CD4+ T cells function and allografts immune protective effects in mice. \u0000 \u0000 \u0000Methods \u0000Mice spleen-derived CD4+ T cells were cultured, purified andtransfected with short hairpin RNA (shRNA) lentiviral vector NLRC5-RNA interference (RNAi)-green fluorescent protein (GFP). The mice allogenic islet transplantation model andskintransplantation model were established, and 24 h prior to transplantation each recipient mice was given CD4+ T cells, NLRC5-RNAi-T cells or equal amount of phosphate buffer (PBS), named T cell group, NLRC5-RNAi group and control group respectively. The survival of islet grafts andskingraftsof transplanted recipients was observed, and on the day 7, the islet graft glucose tolerance, the percentage of T cell subsets in the spleenaswellas related cytokines interleukin (IL)-10 and interferon (IFN)-γwere observed. \u0000 \u0000 \u0000Results \u0000The experimental data demonstratedthat NLRC5-RNAi significantly prolonged the survival timewith a median survival time of (19.0±3.4) days inislet allograftand (15.4±3.8) days inskin allograft, which is significantly longer than control [(11.6±1.9) d, t=5.156, P<0.01] and T cell group [(10.0±1.4) d, t=4.151, P<0.01] inislet allograft, aswellascontrol [(8.0±0.9) d, t=3.375, P<0.05] and T cell group [(7.8±0.8) d, t=3.375, P<0.05] inskin allograft. The results of enzyme linked immunosorbent assay (ELISA) showed that the expression of IL-10 (344.0±4.1) ng/L inisletallograft and (275.9±12.5) ng/L in skinallograftin NLRC5-RNAi group were significantly higher than control group (t=124.141, 20.121, P<0.01) and T cell group (t=32.605, 17.900, P<0.01); the expression of IFN-γ (85.1±6.6) ng/L inisletallograftand (96.6±2.5) ng/L inskinallograftin NLRC5-RNAi group were lower than control group (t=7.633, 7.490, P<0.05) and T cell group (t=10.972, 13.286, P<0.01). Flow cytometric analysis revealed that compared to control group and T cell group, the NLRC5-RNAi group dramatically increased the population of Th2 [(0.190±0.053)%, t=5.220, 5.278, P<0.05] inislet allograft and [(0.130±0.012)%, t=21.060, 9.470, P<0.05] in skin allograft, as well asreduced the population of Th1 [(0.810±0.036)%, t=6.219, 5.276, P<0.05] inislet allograft and [(0.180±0.026)%, t=9.248, 25.324, P<0.05] in skin allograft. \u0000 \u0000 \u0000Conclusion \u0000After inhibiting the expression of NLRC5 gene, the expression of Th2 type cytokines in CD4+ T cells is increased, which prolongs the survival time of grafts and has positive significance for the induction of transplantation tolerance. \u0000 \u0000 \u0000Key words: \u0000NLRC5; Lentivirus transduction; CD4+ T cells; Islet transplantation; Skin transplantation; Immune tolerance","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"33-36"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45494035","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.044
W. Bu, Qinglin Zhao, D. Zhao
Objective To study the clinical significance of early usage of continuous blood purification on the treatment of severe craniocerebral injury. Methods 85 Patients with severe brain injury admitted in our department were randomly divided into continuous blood purification treatment group (experiment group, 42 patients) and conventional treatment group (control group, 43 patients). patients in continuous blood purification treatment group were treated by continuous blood purification and traditional treatment from 1-7 days, traditional treatment group only were treated by the traditional treatment methods. To detect tumor necrosis factor (TNF)-α, neuron specific enolase (NSE) and pNF-H in 1 and 5 day. On the 28th day Apache Ⅱ score, ICU stay time and GCS score of two groups. To detect 6 months survival rate of two groups. Spss15.0 software was used for statistical analysis, The measurement data are subject to T test The survival rate and quality of life comparison between the two groups was compared by χ2 test, P<0.05 was considered statistically significant. Results Incidence of 5 days of TNF-α (4.85±0.45) ng/L (t=6.890, P 0.05). Conclusion Early continuous blood purification treatment improves the survival rate and quality of life of patients with severe brain injury which have higher clinical value. Key words: Continuous blood purification; Sever brain injury; Early treatment
{"title":"Clinical study on early application of continuous blood purification in the treatment of severe craniocerebral injury","authors":"W. Bu, Qinglin Zhao, D. Zhao","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.044","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.044","url":null,"abstract":"Objective \u0000To study the clinical significance of early usage of continuous blood purification on the treatment of severe craniocerebral injury. \u0000 \u0000 \u0000Methods \u000085 Patients with severe brain injury admitted in our department were randomly divided into continuous blood purification treatment group (experiment group, 42 patients) and conventional treatment group (control group, 43 patients). patients in continuous blood purification treatment group were treated by continuous blood purification and traditional treatment from 1-7 days, traditional treatment group only were treated by the traditional treatment methods. To detect tumor necrosis factor (TNF)-α, neuron specific enolase (NSE) and pNF-H in 1 and 5 day. On the 28th day Apache Ⅱ score, ICU stay time and GCS score of two groups. To detect 6 months survival rate of two groups. Spss15.0 software was used for statistical analysis, The measurement data are subject to T test The survival rate and quality of life comparison between the two groups was compared by χ2 test, P<0.05 was considered statistically significant. \u0000 \u0000 \u0000Results \u0000Incidence of 5 days of TNF-α (4.85±0.45) ng/L (t=6.890, P 0.05). \u0000 \u0000 \u0000Conclusion \u0000Early continuous blood purification treatment improves the survival rate and quality of life of patients with severe brain injury which have higher clinical value. \u0000 \u0000 \u0000Key words: \u0000Continuous blood purification; Sever brain injury; Early treatment","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"152-154"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48957098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Effects of astragaloside IV and 5-fluorouracilon on chemosensitization in SW480 tumor transplanted in nude mice and the associated mechanism","authors":"Jianbin Chen, Fang Wang, Weiwei Wang, Guoling Yin, Jianjun Wu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.051","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.051","url":null,"abstract":"","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"176-176"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42996084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.017
Qinghua Zhang, Gui-Zhen Ou, Ruihua Wang, Zhenxing Yu, Jingfeng Liu
Objective To investigate the effect of common bile duct ligation on fibroblast activation in rats with obstructive jaundice. Methods Twenty-four male specific pathogen free (SPF) rats were randomly divided into sham-operated group and experimental group, with 12 rats in each group. All rats underwent common bile duct ligation. Rats The common bile duct in the experimental group was ligated with 3-0 silk thread and cut off. The rats in the two groups were sampled 2 and 7 days after operation. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of transforming growth factor-β1 (TGF-β1). Immunohistochemistry was used to detect the expression of TGF-β1 protein. The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of TGF-β1 gene. Western blotting was used to detect the expression of TGF-β1 protein. Results In sham-operated group, the structure of hepatic lobules was intact, and the hepatic sinuses were normal without congestion. In experimental group, the structure of hepatic lobules was disordered and the hepatic cords were dissociated. The level of serum TGF-β1 in the experimental group at 2nd day [(689.42±58.13) ng/L] and 7th day [(867.53±45.62) ng/L] was significantly higher than that in the sham-operated group [(519.92±32.41) ng/L and (536.50±41.29) ng/L, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 protein in liver tissues of experimental group at 2nd day [(6.53±1.29)%] and 7th day [(8.97±1.85)%] was significantly higher than that of sham-operated group [(1.17±0.24)% and (1.54±0.39)%, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 in liver tissues of experimental group at 2nd day [(0.89±0.14)%] and 7th day [(0.95±0.19)%] was significantly higher than that of sham-operated group [(0.52±1.32)% and (0.31±1.76)%, t=14.693 and 18.316, P<0.05]. The gray value of TGF-β1 protein expression in liver tissue of experimental group at 2nd day after operation (0.62±0.13) and 7th day after operation (0.78±0.10) was significantly higher than that of sham-operated group (0.21±0.05 and 0.23±0.06, t=10.197 and 16.337, P<0.05). Conclusion The expression of TGF-β1 in rats with obstructive jaundice induced by common bile duct ligation is significantly up-regulated, and the proliferation of biliary duct epithelial cells induces fibroblast activation. Key words: Common bile duct ligation; Obstructive jaundice; Fibroblast activation
{"title":"Fibroblast activation induced by common bile duct ligation in rats with obstructive jaundice","authors":"Qinghua Zhang, Gui-Zhen Ou, Ruihua Wang, Zhenxing Yu, Jingfeng Liu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.017","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.017","url":null,"abstract":"Objective \u0000To investigate the effect of common bile duct ligation on fibroblast activation in rats with obstructive jaundice. \u0000 \u0000 \u0000Methods \u0000Twenty-four male specific pathogen free (SPF) rats were randomly divided into sham-operated group and experimental group, with 12 rats in each group. All rats underwent common bile duct ligation. Rats The common bile duct in the experimental group was ligated with 3-0 silk thread and cut off. The rats in the two groups were sampled 2 and 7 days after operation. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of transforming growth factor-β1 (TGF-β1). Immunohistochemistry was used to detect the expression of TGF-β1 protein. The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the expression of TGF-β1 gene. Western blotting was used to detect the expression of TGF-β1 protein. \u0000 \u0000 \u0000Results \u0000In sham-operated group, the structure of hepatic lobules was intact, and the hepatic sinuses were normal without congestion. In experimental group, the structure of hepatic lobules was disordered and the hepatic cords were dissociated. The level of serum TGF-β1 in the experimental group at 2nd day [(689.42±58.13) ng/L] and 7th day [(867.53±45.62) ng/L] was significantly higher than that in the sham-operated group [(519.92±32.41) ng/L and (536.50±41.29) ng/L, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 protein in liver tissues of experimental group at 2nd day [(6.53±1.29)%] and 7th day [(8.97±1.85)%] was significantly higher than that of sham-operated group [(1.17±0.24)% and (1.54±0.39)%, t=8.822 and 18.635, P<0.05]. The expression of TGF-β1 in liver tissues of experimental group at 2nd day [(0.89±0.14)%] and 7th day [(0.95±0.19)%] was significantly higher than that of sham-operated group [(0.52±1.32)% and (0.31±1.76)%, t=14.693 and 18.316, P<0.05]. The gray value of TGF-β1 protein expression in liver tissue of experimental group at 2nd day after operation (0.62±0.13) and 7th day after operation (0.78±0.10) was significantly higher than that of sham-operated group (0.21±0.05 and 0.23±0.06, t=10.197 and 16.337, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000The expression of TGF-β1 in rats with obstructive jaundice induced by common bile duct ligation is significantly up-regulated, and the proliferation of biliary duct epithelial cells induces fibroblast activation. \u0000 \u0000 \u0000Key words: \u0000Common bile duct ligation; Obstructive jaundice; Fibroblast activation","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"60-62"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47648342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.030
Ximeng Ji, H. Zhuang, Yiwen Zhang, Jincan Zhang, Bu Shoushan
Objective To construct an artificial nerve graft as an alternative for autologous nerve grafting. Methods We fabricated an innovative tissue engineered nerve graft (TENG), which consists of a commercial bilayer collagen membrane (Bio-Gide)-based nerve scaffold and bone marrow mesenchymal stem cells (BMSCs) serving as support cells to bridge a 10 mm long sciatic nerve defect in rats, The 10 mm gap was left empty after resection in the blank group. In the other three groups, the sciatic nerve defect was, respectively, bridged with plain collagen tube (CT group), a collagen tube filled with BMSC suspension (C+ CT group), or reversed transected nerve segment (AG group). Results Histological and functional assessments showed that the developed TENG facilitated nerve regeneration (SFI: CT, -92.490±1.836; C+ CT, -70.010±7.805; AG, -70.130±8.744; F= 17.850, P 0.05) and better outcomes than the plain collagen tube (P<0.05). Conclusion These results manifested that the bilayer collagen conduit loaded BMSCs could be applied as a new biodegradable artificial nerve guide for nerve tissue engineering. Key words: Tissue engineer; Nerve graft; Bone marrow mesenchymal stem cells; Bilayer collagen conduit scaffold; Sciatic nerve injury; Peripheral nerve regeneration
{"title":"Repairing rat sciatic nerve defects via transplanting bilayer collagen conduit scaffolds carrying bone marrow mesenchymal stem cells","authors":"Ximeng Ji, H. Zhuang, Yiwen Zhang, Jincan Zhang, Bu Shoushan","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.030","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.030","url":null,"abstract":"Objective \u0000To construct an artificial nerve graft as an alternative for autologous nerve grafting. \u0000 \u0000 \u0000Methods \u0000We fabricated an innovative tissue engineered nerve graft (TENG), which consists of a commercial bilayer collagen membrane (Bio-Gide)-based nerve scaffold and bone marrow mesenchymal stem cells (BMSCs) serving as support cells to bridge a 10 mm long sciatic nerve defect in rats, The 10 mm gap was left empty after resection in the blank group. In the other three groups, the sciatic nerve defect was, respectively, bridged with plain collagen tube (CT group), a collagen tube filled with BMSC suspension (C+ CT group), or reversed transected nerve segment (AG group). \u0000 \u0000 \u0000Results \u0000Histological and functional assessments showed that the developed TENG facilitated nerve regeneration (SFI: CT, -92.490±1.836; C+ CT, -70.010±7.805; AG, -70.130±8.744; F= 17.850, P 0.05) and better outcomes than the plain collagen tube (P<0.05). \u0000 \u0000 \u0000Conclusion \u0000These results manifested that the bilayer collagen conduit loaded BMSCs could be applied as a new biodegradable artificial nerve guide for nerve tissue engineering. \u0000 \u0000 \u0000Key words: \u0000Tissue engineer; Nerve graft; Bone marrow mesenchymal stem cells; Bilayer collagen conduit scaffold; Sciatic nerve injury; Peripheral nerve regeneration","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"105-108"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46777948","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.014
Zengguang Zhang, Heyuan Sun, Fengbiao Wang
Objective To study the mechanism of CC chemokine ligand-5 promoting macrophage infiltration in early stage of hepatic ischemia-reperfusion injury. Methods The adult male C57BL/6 wild type and CC chemokine ligand-5 (CCL5) deficient mice were used for the experiment. The mice were kept in captivity at (23±2) ℃, and were exposed to light and dark cycles for 12 hours to obtain food and water freely. The mice were randomly divided into 4 groups. Sham operation group (sham operation group mice experienced ischemia-reperfusion model scheme, but no vascular occlusion, n=10). In the ischemia-reperfusion model group (mice were anesthetized with isoflurane and underwent midline laparotomy, and a noninvasive clamp was placed on the portal, hepatic artery and bile duct to interrupt the blood supply of the left lateral lobe and middle lobe. After 60 minutes of partial liver ischemia, the clamp was removed to start reperfusion, n=10). CCL5 antagonists + model group (before the start of reperfusion, the CCL5 antagonists + model group received a single intravenous injection of the CCR5 receptor antagonist maladono, n=10); CCL5 defective group (CCL5 defective mice, n=10). Alanine aminotransferase (ALT) content and myeloperoxidase (MPO) activity were measured by vtros dt60 Ⅱ chemical system. The expression of ALT mRNA and MPO mRNA was detected by eal-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The number of macrophages was detected by flow cytometry. The infiltration of macrophages into the lung was detected by histology and immunohistochemistry. The protein expression of related inflammatory factors was detected by Western blotting. Results Compared with the sham operation group, ALT content [(521.36±20.65) vs. (4 126.55±326.68), F=16.237, P 0.05). Compared with CCL5 deficiency group, the number of CD45+ in ischemia-reperfusion model group increased at 24 hours [(2.91±0.23) vs. (1.65±0.17), F=16.311, P<0.05]. At 1, 2, 8 and 24 hours after reperfusion, the number of CD68+ cells in the ischemia-reperfusion model group was higher than that in the sham operation group [(94.62±8.55) vs. (23.68±3.11), F=16.352, P<0.05]. However, in the early stage of ischemia-reperfusion, CCL5 deficiency group was lower than that of ischemia-reperfusion model group [(76.38±6.77) vs. (94.62±8.55), F=16.352, P<0.05]. Compared with the sham operation group, the protein expression of tumor necrosis factor-α (TNF-α) [(1.12±0.23) vs. (3.24±0.42), F=11.352, P<0.05], interleukin (IL)-1 β [(1.03±0.08) vs. (2.87±0.35), F=12.452, P<0.05], IL-6 [(0.95±0.02) vs. (2.58±0.21), F=15.652, P<0.05] in the ischemia-reperfusion model group increased. However, the expression of TNF - α [(3.24±0.42) vs. (1.35±0.14), F=14.252, P<0.05], IL-1 β [(2.87±0.35) vs. (1.22±0.15), F=13.723, P<0.05], IL-6 [(2.58±0.21) vs. (1.35±0.26), F=14.312, P<0.05] in CCL5 antagonist + model group was lower than that in ischemia-reperfusion model group. Conclusion CCL5 can promote
{"title":"CC chemokine ligand-5 promotes macrophage infiltration in early stage of hepatic ischemia-reperfusion injury","authors":"Zengguang Zhang, Heyuan Sun, Fengbiao Wang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.014","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.014","url":null,"abstract":"Objective \u0000To study the mechanism of CC chemokine ligand-5 promoting macrophage infiltration in early stage of hepatic ischemia-reperfusion injury. \u0000 \u0000 \u0000Methods \u0000The adult male C57BL/6 wild type and CC chemokine ligand-5 (CCL5) deficient mice were used for the experiment. The mice were kept in captivity at (23±2) ℃, and were exposed to light and dark cycles for 12 hours to obtain food and water freely. The mice were randomly divided into 4 groups. Sham operation group (sham operation group mice experienced ischemia-reperfusion model scheme, but no vascular occlusion, n=10). In the ischemia-reperfusion model group (mice were anesthetized with isoflurane and underwent midline laparotomy, and a noninvasive clamp was placed on the portal, hepatic artery and bile duct to interrupt the blood supply of the left lateral lobe and middle lobe. After 60 minutes of partial liver ischemia, the clamp was removed to start reperfusion, n=10). CCL5 antagonists + model group (before the start of reperfusion, the CCL5 antagonists + model group received a single intravenous injection of the CCR5 receptor antagonist maladono, n=10); CCL5 defective group (CCL5 defective mice, n=10). Alanine aminotransferase (ALT) content and myeloperoxidase (MPO) activity were measured by vtros dt60 Ⅱ chemical system. The expression of ALT mRNA and MPO mRNA was detected by eal-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The number of macrophages was detected by flow cytometry. The infiltration of macrophages into the lung was detected by histology and immunohistochemistry. The protein expression of related inflammatory factors was detected by Western blotting. \u0000 \u0000 \u0000Results \u0000Compared with the sham operation group, ALT content [(521.36±20.65) vs. (4 126.55±326.68), F=16.237, P 0.05). Compared with CCL5 deficiency group, the number of CD45+ in ischemia-reperfusion model group increased at 24 hours [(2.91±0.23) vs. (1.65±0.17), F=16.311, P<0.05]. At 1, 2, 8 and 24 hours after reperfusion, the number of CD68+ cells in the ischemia-reperfusion model group was higher than that in the sham operation group [(94.62±8.55) vs. (23.68±3.11), F=16.352, P<0.05]. However, in the early stage of ischemia-reperfusion, CCL5 deficiency group was lower than that of ischemia-reperfusion model group [(76.38±6.77) vs. (94.62±8.55), F=16.352, P<0.05]. Compared with the sham operation group, the protein expression of tumor necrosis factor-α (TNF-α) [(1.12±0.23) vs. (3.24±0.42), F=11.352, P<0.05], interleukin (IL)-1 β [(1.03±0.08) vs. (2.87±0.35), F=12.452, P<0.05], IL-6 [(0.95±0.02) vs. (2.58±0.21), F=15.652, P<0.05] in the ischemia-reperfusion model group increased. However, the expression of TNF - α [(3.24±0.42) vs. (1.35±0.14), F=14.252, P<0.05], IL-1 β [(2.87±0.35) vs. (1.22±0.15), F=13.723, P<0.05], IL-6 [(2.58±0.21) vs. (1.35±0.26), F=14.312, P<0.05] in CCL5 antagonist + model group was lower than that in ischemia-reperfusion model group. \u0000 \u0000 \u0000Conclusion \u0000CCL5 can promote ","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"48-51"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46563304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Objective To explore the expression and significance of armadillo repeat-containing X-linked protein 3 (ALEX3) in papillary thyroid carcinoma. Methods From June 2018 to September 2018 a total of 60 cases (10 males and 50 females) of papillary thyroid carcinoma in the First Affiliated Hospital of Zhengzhou University were collected. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were used to detect the expression levels of ALEX3 in papillary thyroid carcinoma tissues and normal thyroid tissues, respectively. The median value of the ALEX3 expression level is used as the truncated value, 60 patients with papillary thyroid cancer were divided into the ALEX3 high-expression group and the ALEX3 low-expression group. The t test was used to analyze the difference in ALEX3 expression level between thyroid papillary carcinoma tissues and normal thyroid tissues, and the χ2 test was used to analyze the correlation between ALEX3 mRNA and clinicopathological features in papillary thyroid carcinoma. Results The relative expression value of ALEX3 mRNA in papillary thyroid carcinoma tissues (2.125±0.755) was significantly higher than that in normal thyroid tissues (1.156±0.426, t=8.663, P<0.05). The difference was statistically significant.Expression level of ALEX3 mRNA was closely related to TNM pathological stage and lymph node metastasis (χ2=5.963, 15.152, P<0.05). The relative expression value of ALEX3 protein in papillary thyroid carcinoma tissues (0.332±0.145) was significantly higher than that in normal thyroid tissues (0.204±0.132, t=2.252, P<0.05), The difference was statistically significant.. Conclusion ALEX3 was significantly over-expressed in thyroid papillary carcinoma, and might play an important role in its occurrence and progression. Key words: Armadillo repeat-containing X-linked protein 3; Papillary thyroid carcinoma
{"title":"Expression and clinical significance of armadillo repeat-containing X-linked protein 3 in papillary thyroid carcinoma","authors":"Jianhua Li, Zhen Deng, Pengfei Xu, Wenping Xue, L. Fu, Shouhua Zheng, Xinguang Qiu","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.037","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.037","url":null,"abstract":"Objective \u0000To explore the expression and significance of armadillo repeat-containing X-linked protein 3 (ALEX3) in papillary thyroid carcinoma. \u0000 \u0000 \u0000Methods \u0000From June 2018 to September 2018 a total of 60 cases (10 males and 50 females) of papillary thyroid carcinoma in the First Affiliated Hospital of Zhengzhou University were collected. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting were used to detect the expression levels of ALEX3 in papillary thyroid carcinoma tissues and normal thyroid tissues, respectively. The median value of the ALEX3 expression level is used as the truncated value, 60 patients with papillary thyroid cancer were divided into the ALEX3 high-expression group and the ALEX3 low-expression group. The t test was used to analyze the difference in ALEX3 expression level between thyroid papillary carcinoma tissues and normal thyroid tissues, and the χ2 test was used to analyze the correlation between ALEX3 mRNA and clinicopathological features in papillary thyroid carcinoma. \u0000 \u0000 \u0000Results \u0000The relative expression value of ALEX3 mRNA in papillary thyroid carcinoma tissues (2.125±0.755) was significantly higher than that in normal thyroid tissues (1.156±0.426, t=8.663, P<0.05). The difference was statistically significant.Expression level of ALEX3 mRNA was closely related to TNM pathological stage and lymph node metastasis (χ2=5.963, 15.152, P<0.05). The relative expression value of ALEX3 protein in papillary thyroid carcinoma tissues (0.332±0.145) was significantly higher than that in normal thyroid tissues (0.204±0.132, t=2.252, P<0.05), The difference was statistically significant.. \u0000 \u0000 \u0000Conclusion \u0000ALEX3 was significantly over-expressed in thyroid papillary carcinoma, and might play an important role in its occurrence and progression. \u0000 \u0000 \u0000Key words: \u0000Armadillo repeat-containing X-linked protein 3; Papillary thyroid carcinoma","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"128-130"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47064405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.054
C. Luo, Zeyuan Yu, Gang Chen, Long Li, Ruiying Luo, Z. Jiao
{"title":"Expression of ubiquitin-conjugating enzyme E2T in gastric cancer and para-carcinoma tissues","authors":"C. Luo, Zeyuan Yu, Gang Chen, Long Li, Ruiying Luo, Z. Jiao","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.054","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.054","url":null,"abstract":"","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"179-180"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45061753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2020-01-08DOI: 10.3760/CMA.J.ISSN.1001-9030.2020.01.005
Chuan-jiang Liu, Peng Xia, Pan Liu, Wen-ping Zhou, Q. Fu, T. Qin, Hongwei Zhang
Objective To observe the expression of microRNA (miRNA, miR)-15b in pancreatic cancer (PC) cells and its effect on proliferation, migration and apoptosis of pancreatic cancer cell lines. Methods Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect miR-15b expression in four pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) and human normal pancreatic ductal epithelial cell line (HPDE6c7). CFPAC-1 and PANC-1 cells stably transfected with miR-15b short hairpin RNA (shRNA) were used as experimental group, and CFPAC-1 and PANC-1 cells which normally expressed miR-15b were transfected with negative control (NC) as control group. The proliferation of CFPAC-1 and PANC-1 cells was measured by cell counting kit-8 (CCK-8) proliferation assay after transfection. The migration ability of CFPAC-1 and PANC-1 cells was measured by Transwell migration assay. The apoptosis of CFPAC-1 and PANC-1 cells was examined by flow cytometry. Results The expression levels of miR-15b in 4 pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) were 3.63±1.02, 5.28±0.76, 6.72±1.39 and 4.68±1.56 respectively, which were significantly higher than that (1.08±0.23) in HPDE6c7 cells (t=9.944, 10.326, 12.013, 15.877, P<0.01). The cell proliferation ability of CFPAC-1 cells in miR-15b low expression group at 2, 3 and 4 d (0.51±0.04, 0.94±0.06, 1.25±0.15) was significantly reduced correspondingly as compared with that of CFPAC-1 cells in control group (0.68±0.06, 1.21±0.09, 1.95±0.12) (t=8.256, 6.350, 7.002, P<0.05). The cell proliferation ability of PANC-1 cells in miR-15b low expression group at 2, 3 and 4 d(0.48±0.05, 0.76±0.13, 1.44±0.25) was significantly decreased correspondingly as compared with that in the control group (0.58±0.07, 0.98±0.13, 1.86±0.14) (t=9.988, 10.022, 13.050, P<0.01). The number of CFPAC-1 and PANC-1 cells migrating through the Transwell chamber in the miR-15b low-expression group (38.47±4.69 and 47.83±12.47) was significantly lower than that in the control group (62.39±7.39 and 68.97±8.44) (t=5.798, 8.465, P<0.05). The apoptosis rate of CFPAC-1 and PANC-1 cells in the miR-15b low-expression group [(14.68±3.21)% and (11.57±2.52)%] was significantly higher corresponding than that in the control group [(4.29±1.42)% and (4.73±0.38)%] (t=6.350, 8.666, P<0.05). Conclusion MiR-15b is highly expressed in PC cells. Low expression of miR-15b can decrease the proliferation and migration of PC cells and promote apoptosis of PC cells. Key words: MicroRNA-15b; Proliferation; Migration; Apoptosis
{"title":"Function of microRNA-15b in proliferation, migration and apoptosis of pancreatic cancer cells","authors":"Chuan-jiang Liu, Peng Xia, Pan Liu, Wen-ping Zhou, Q. Fu, T. Qin, Hongwei Zhang","doi":"10.3760/CMA.J.ISSN.1001-9030.2020.01.005","DOIUrl":"https://doi.org/10.3760/CMA.J.ISSN.1001-9030.2020.01.005","url":null,"abstract":"Objective \u0000To observe the expression of microRNA (miRNA, miR)-15b in pancreatic cancer (PC) cells and its effect on proliferation, migration and apoptosis of pancreatic cancer cell lines. \u0000 \u0000 \u0000Methods \u0000Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) was used to detect miR-15b expression in four pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) and human normal pancreatic ductal epithelial cell line (HPDE6c7). CFPAC-1 and PANC-1 cells stably transfected with miR-15b short hairpin RNA (shRNA) were used as experimental group, and CFPAC-1 and PANC-1 cells which normally expressed miR-15b were transfected with negative control (NC) as control group. The proliferation of CFPAC-1 and PANC-1 cells was measured by cell counting kit-8 (CCK-8) proliferation assay after transfection. The migration ability of CFPAC-1 and PANC-1 cells was measured by Transwell migration assay. The apoptosis of CFPAC-1 and PANC-1 cells was examined by flow cytometry. \u0000 \u0000 \u0000Results \u0000The expression levels of miR-15b in 4 pancreatic cancer cell lines (SW1990, CFPAC-1, PANC-1, BxPC-3) were 3.63±1.02, 5.28±0.76, 6.72±1.39 and 4.68±1.56 respectively, which were significantly higher than that (1.08±0.23) in HPDE6c7 cells (t=9.944, 10.326, 12.013, 15.877, P<0.01). The cell proliferation ability of CFPAC-1 cells in miR-15b low expression group at 2, 3 and 4 d (0.51±0.04, 0.94±0.06, 1.25±0.15) was significantly reduced correspondingly as compared with that of CFPAC-1 cells in control group (0.68±0.06, 1.21±0.09, 1.95±0.12) (t=8.256, 6.350, 7.002, P<0.05). The cell proliferation ability of PANC-1 cells in miR-15b low expression group at 2, 3 and 4 d(0.48±0.05, 0.76±0.13, 1.44±0.25) was significantly decreased correspondingly as compared with that in the control group (0.58±0.07, 0.98±0.13, 1.86±0.14) (t=9.988, 10.022, 13.050, P<0.01). The number of CFPAC-1 and PANC-1 cells migrating through the Transwell chamber in the miR-15b low-expression group (38.47±4.69 and 47.83±12.47) was significantly lower than that in the control group (62.39±7.39 and 68.97±8.44) (t=5.798, 8.465, P<0.05). The apoptosis rate of CFPAC-1 and PANC-1 cells in the miR-15b low-expression group [(14.68±3.21)% and (11.57±2.52)%] was significantly higher corresponding than that in the control group [(4.29±1.42)% and (4.73±0.38)%] (t=6.350, 8.666, P<0.05). \u0000 \u0000 \u0000Conclusion \u0000MiR-15b is highly expressed in PC cells. Low expression of miR-15b can decrease the proliferation and migration of PC cells and promote apoptosis of PC cells. \u0000 \u0000 \u0000Key words: \u0000MicroRNA-15b; Proliferation; Migration; Apoptosis","PeriodicalId":10065,"journal":{"name":"中华实验外科杂志","volume":"37 1","pages":"19-21"},"PeriodicalIF":0.0,"publicationDate":"2020-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49337781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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