AIM： To establish and validate the mutation testing for identification and characterization of hereditary non-polyposis colorectal cancer （HNPCC） in suspected Chinese patients. METHODS： Five independent Chinese ki...AIM： To establish and validate the mutation testing for identification and characterization of hereditary non-polyposis colorectal cancer （HNPCC） in suspected Chinese patients. METHODS： Five independent Chinese kindreds with HNPCC fulfilling the classical Amsterdam criteria were collected. Genomic DNA was extracted after informed consent was obtained. The coding region of hMSH2 and hMLH1 genes was detected by polymerase chain reaction （PCR） and denaturing high-performance liquid chromatography （DHPLC）. Mutations identified in the proband by DHPLC were directly sequenced using a 377 DNA sequencer, analyzed with a basic local alignment tool （BLAST）, and tested in the corresponding family members by direct DNA sequencing. RESULTS： Mutations were identified in two Chinese HNPCC kindreds. One was the missense mutation of hMSH2 c.1808A→G resulting in Asp 603 Gly identified in the proband of the fifth HNPCC （HNPCCS） kindred. In the HNP5 kindred, three family members were found to have this mutation and two of them had colorectal cancer. The other mutation of hMLH1 c.1882A→G was identified in the HNP2 kindred＇s proband, which might be the nonsense mutation analyzed by BLAST. CONCLUSION： Pedigree investigation and mutation testing of hMSH2 and hMLH1 are the practical methods to identify high-risk HNPCC patients in China.展开更多
目的:通过观察健脾活血方对胃癌前病变大鼠胃黏膜组织中CD44V6、MLH1及MSH2表达的影响,探讨健脾活血方对其干预的作用机制.方法:除正常组外,其他大鼠采用以N-甲基-N-硝基-N-亚硝基胍(N-methyl-N-nitro N-nitrosoguanidine,MNNG)为主同...目的:通过观察健脾活血方对胃癌前病变大鼠胃黏膜组织中CD44V6、MLH1及MSH2表达的影响,探讨健脾活血方对其干预的作用机制.方法:除正常组外,其他大鼠采用以N-甲基-N-硝基-N-亚硝基胍(N-methyl-N-nitro N-nitrosoguanidine,MNNG)为主同时配合0.3g/L雷尼替丁、400 mL/L乙醇及饥饱失常的多因素造模法建立胃癌前病变动物模型.将造模成功的40只大鼠随机分为模型组(0.9%氯化钠溶液)、胃复春组(0.86 g/kg)、健脾活血方高、中、低剂量组(32、16、8 g/kg),每组8只,每组每天给予等量(10 mL/kg)的不同药物灌胃一次,连续10 wk.实验末处死大鼠,给予相应处理后,快速免疫组织化学检测CD44V6、MLH1及MSH2表达情况.结果:模型组CD44V6表达与正常组相比明显升高(5.12±1.96 vs 0.25±0.46,P<0.01);健脾活血方高、中剂量组CD44V6表达与模型组相比均明显降低(2.25±0.71,3.25±0.31vs 5.12±1.96,P<0.01或P<0.05),低剂量组C D44V6表达与模型组比较差异无统计学意义(P>0.05);健脾活血方高剂量组CD44V6表达与胃复春组相比明显降低(2.25±0.71 vs4.62±1.19,P<0.01),中、低剂量组CD44V6表达与胃复春组比较差异无统计学意义(P>0.05).模型组MLH1、MSH2表达与正常组相比均明显降低(3.75±1.04 vs 8.00±0.926;3.62±1.69 vs 7.25±2.12,P<0.01);健脾活血方高、中、低剂量组MLH1、MSH2表达与模型组相比均明显升高(6.50±0.93,5.25±1.49,5.12±1.25 vs 3.75±1.04;6.62±2.13,6.00±1.51,5.50±1.41 vs 3.62±1.69,P<0.01或P<0.05);健脾活血方高剂量组MLH1表达与胃复春组相比明显升高(6.50±0.93 vs 4.88±1.25,P<0.05),中、低剂量组MLH1及高、中、低剂量组MSH2表达与胃复春组比较差异无统计学意义(P>0.05).结论:健脾活血方可通过降低CD44V6表达,上调MLH1、MSH2表达,减少细胞的非正常侵袭和转移,增强基因的错配修复功能,减少细胞的异常增殖和分化,发挥对大鼠胃癌前病变的治疗作用.展开更多
对四川猪链球菌2型(S.suis 2)强毒株05ZYH33溶血素样蛋白(hemolysin-like protein,HLP)编码基因hlp进行序列信息分析、分子克隆表达及溶血活性检测,深入探讨HLP的致病机制.用Blast和Clustal X程序对HLP进行基因同源性分析,用Signal P 4....对四川猪链球菌2型(S.suis 2)强毒株05ZYH33溶血素样蛋白(hemolysin-like protein,HLP)编码基因hlp进行序列信息分析、分子克隆表达及溶血活性检测,深入探讨HLP的致病机制.用Blast和Clustal X程序对HLP进行基因同源性分析,用Signal P 4.1和TMHMM Server 2.0分析HLP氨基酸序列.设计合成hlp引物进行PCR扩增,先后将hlp克隆入p MD-18T载体和p ET30a表达载体中,构建p ET30a::hly原核表达质粒.将重组质粒转化至E.coli BL21中诱导表达,并对重组HLP蛋白进行纯化和溶血活性检测.同源性分析表明HLP与多种具有溶血活性的蛋白相似性较高.氨基酸序列分析发现HLP具有信号肽和跨膜区,且由DUF21、CBS和Cor CHly C 3部分组成.序列测定显示hly片段全长1 335 bp,编码445个氨基酸.重组质粒经诱导表达和纯化后电泳显示,HLP分子量约为70 k Da,具有溶血活性.结果表明,HLP是一个膜结合蛋白,不同于能够分泌到细胞外的溶血素Suilysin.重组HLP具有一定的溶血效价,此可能与致病相关.展开更多
Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and af...Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.展开更多
基金The Special Funds of China Education Ministry for Returnees, No. 2003-14
文摘AIM： To establish and validate the mutation testing for identification and characterization of hereditary non-polyposis colorectal cancer （HNPCC） in suspected Chinese patients. METHODS： Five independent Chinese kindreds with HNPCC fulfilling the classical Amsterdam criteria were collected. Genomic DNA was extracted after informed consent was obtained. The coding region of hMSH2 and hMLH1 genes was detected by polymerase chain reaction （PCR） and denaturing high-performance liquid chromatography （DHPLC）. Mutations identified in the proband by DHPLC were directly sequenced using a 377 DNA sequencer, analyzed with a basic local alignment tool （BLAST）, and tested in the corresponding family members by direct DNA sequencing. RESULTS： Mutations were identified in two Chinese HNPCC kindreds. One was the missense mutation of hMSH2 c.1808A→G resulting in Asp 603 Gly identified in the proband of the fifth HNPCC （HNPCCS） kindred. In the HNP5 kindred, three family members were found to have this mutation and two of them had colorectal cancer. The other mutation of hMLH1 c.1882A→G was identified in the HNP2 kindred＇s proband, which might be the nonsense mutation analyzed by BLAST. CONCLUSION： Pedigree investigation and mutation testing of hMSH2 and hMLH1 are the practical methods to identify high-risk HNPCC patients in China.
文摘目的:通过观察健脾活血方对胃癌前病变大鼠胃黏膜组织中CD44V6、MLH1及MSH2表达的影响,探讨健脾活血方对其干预的作用机制.方法:除正常组外,其他大鼠采用以N-甲基-N-硝基-N-亚硝基胍(N-methyl-N-nitro N-nitrosoguanidine,MNNG)为主同时配合0.3g/L雷尼替丁、400 mL/L乙醇及饥饱失常的多因素造模法建立胃癌前病变动物模型.将造模成功的40只大鼠随机分为模型组(0.9%氯化钠溶液)、胃复春组(0.86 g/kg)、健脾活血方高、中、低剂量组(32、16、8 g/kg),每组8只,每组每天给予等量(10 mL/kg)的不同药物灌胃一次,连续10 wk.实验末处死大鼠,给予相应处理后,快速免疫组织化学检测CD44V6、MLH1及MSH2表达情况.结果:模型组CD44V6表达与正常组相比明显升高(5.12±1.96 vs 0.25±0.46,P<0.01);健脾活血方高、中剂量组CD44V6表达与模型组相比均明显降低(2.25±0.71,3.25±0.31vs 5.12±1.96,P<0.01或P<0.05),低剂量组C D44V6表达与模型组比较差异无统计学意义(P>0.05);健脾活血方高剂量组CD44V6表达与胃复春组相比明显降低(2.25±0.71 vs4.62±1.19,P<0.01),中、低剂量组CD44V6表达与胃复春组比较差异无统计学意义(P>0.05).模型组MLH1、MSH2表达与正常组相比均明显降低(3.75±1.04 vs 8.00±0.926;3.62±1.69 vs 7.25±2.12,P<0.01);健脾活血方高、中、低剂量组MLH1、MSH2表达与模型组相比均明显升高(6.50±0.93,5.25±1.49,5.12±1.25 vs 3.75±1.04;6.62±2.13,6.00±1.51,5.50±1.41 vs 3.62±1.69,P<0.01或P<0.05);健脾活血方高剂量组MLH1表达与胃复春组相比明显升高(6.50±0.93 vs 4.88±1.25,P<0.05),中、低剂量组MLH1及高、中、低剂量组MSH2表达与胃复春组比较差异无统计学意义(P>0.05).结论:健脾活血方可通过降低CD44V6表达,上调MLH1、MSH2表达,减少细胞的非正常侵袭和转移,增强基因的错配修复功能,减少细胞的异常增殖和分化,发挥对大鼠胃癌前病变的治疗作用.
文摘对四川猪链球菌2型(S.suis 2)强毒株05ZYH33溶血素样蛋白(hemolysin-like protein,HLP)编码基因hlp进行序列信息分析、分子克隆表达及溶血活性检测,深入探讨HLP的致病机制.用Blast和Clustal X程序对HLP进行基因同源性分析,用Signal P 4.1和TMHMM Server 2.0分析HLP氨基酸序列.设计合成hlp引物进行PCR扩增,先后将hlp克隆入p MD-18T载体和p ET30a表达载体中,构建p ET30a::hly原核表达质粒.将重组质粒转化至E.coli BL21中诱导表达,并对重组HLP蛋白进行纯化和溶血活性检测.同源性分析表明HLP与多种具有溶血活性的蛋白相似性较高.氨基酸序列分析发现HLP具有信号肽和跨膜区,且由DUF21、CBS和Cor CHly C 3部分组成.序列测定显示hly片段全长1 335 bp,编码445个氨基酸.重组质粒经诱导表达和纯化后电泳显示,HLP分子量约为70 k Da,具有溶血活性.结果表明,HLP是一个膜结合蛋白,不同于能够分泌到细胞外的溶血素Suilysin.重组HLP具有一定的溶血效价,此可能与致病相关.
基金supported by American Diabetes Association,American Heart Association,NIH NIEHS,NIH NIA,NIH NINDS,and NIH ARRA
文摘Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.
