SHP-2和转化生长因子β在中老年患者宫颈鳞癌组织中的表达

目的 观察宫颈癌患者癌组织SHP-2和转化生长因子β的表达及其与宫颈癌分型的关系.方法 纳入2009年7月至2012年4月在沈阳医学院奉天医院妇产科住院治疗的120例宫颈鳞癌患者、80例宫颈上皮内肿瘤患者及30例宫颈正常患者,取宫颈组织,免疫组化染色检测宫颈组织中SHP-2和转化生长因子β的表达;实时RT-PCR和Western印迹检测宫颈组织SHP-2 mRNA和蛋白的表达.结果 免疫组化染色显示,SHP-2和转化生长因子β主要表达在胞浆,且其表达均在宫颈鳞癌组织中最高,其次是宫颈上皮内肿瘤组织,在正常宫颈仅有少量表达.实时RT-PCR和Western印迹检测结果与免疫组化结果一致.宫颈鳞癌组织SHP-2 mRNA和蛋白的表达明显高于宫颈上皮内肿瘤组织和正常宫颈组织(P＜0.05);与正常宫颈组织比较,宫颈上皮内肿瘤组织SHP-2 mRNA和蛋白的表达明显增高(P＜0.05).结论 SHP-2和转化生长因子β共同参与宫颈癌的形成,并与宫颈癌的分型有关.     

RhoC基因mRNA在食管鳞癌中的表达及其意义

目的:探讨RhoC基因在食管鳞癌组织中的表达情况及其与食管鳞癌发生、发展的关系.方法:采用半定量RT-PCR和原位杂交方法检测62例食管鳞癌、31例癌旁不典型增生组织及62例正常食管黏膜组织中RhoC mRNA的相对表达量及细胞定位.结果:食管鳞癌组织中RhoC mRNA表达与癌的组织学分级、浸润深度及淋巴结转移密切相关(P <0.05); 在食管鳞癌癌变过程中,RT-PCR检测RhoC mRNA在癌组织、癌旁不典型增生组织及正常黏膜组织中的表达量依次降低, 分别为0.902±0.119、0.731±0.065、0.653±0.069, 组间比较有明显差异(H= 99.629, P <0.01); 原位杂交检测RhoC转录本主要位于细胞质中, 其在癌组织、癌旁不典型增生组织及正常黏膜组织中的表达率依次降低, 分别为80.6%(50/62)、32.3%(10/31)、21.0%(13/62), 组间比较有明显差异(χ2 =47.735, P <0.01), 两种方法检测的RhoC mRNA表达具有一致性.结论:RhoC mRNA在食管鳞癌组织中显著增加, 并与食管鳞癌生物学行为关系密切, 提示RhoC mRNA过表达与食管鳞癌的发生、发展有关, RhoC mRNA可作为食管鳞癌早期诊断和判断预后的辅助指标.     

RhoC、Ki-67、Bcl-2在胃癌发生发展中的表达及其相关性

目的 研究胃癌组织和正常胃黏膜中RhoC、Ki-67、Bcl-2的表达情况,探讨它们在胃癌发生发展中的作用及其相关性.方法 应用免疫组化SABC法检测54例胃癌标本及20例正常胃黏膜中RhoC、Ki-67、Bcl-2的表达情况.结果 ①胃癌组织RhoC蛋白、Ki-67蛋白及Bcl-2蛋白阳性表达率高于正常胃黏膜组织(P＜0.05).②随着胃癌浸润深度的增加、分化程度的降低、淋巴结转移的出现RhoC及 Ki-67阳性表达率升高(P＜0.05).③胃癌组织Bcl-2蛋白阳性表达率与胃癌的分化程度具有相关性(P＜0.05).④在胃癌组织中,RhoC与Ki-67的表达呈正相关(r=0.390,P＜0.05),均呈升高趋势;但是二者与Bcl-2均无明显相关(P＞0.05).结论 RhoC、Ki-67、Bcl-2均参与胃癌的发生和发展,三者可成为判断胃癌恶性程度有价值的指标,也可成为预后评估的指标.     

宫颈癌组织中Sox-2的表达变化及意义

目的观察宫颈鳞癌组织中转录因子Sox-2的表达变化,并探讨其临床意义。方法采用免疫组化法检测84例宫颈鳞癌组织、66例宫颈CIN组织、22例正常宫颈组织中的Sox-2蛋白,采用RT-PCR法测定Sox-2 mRNA。结果宫颈癌组织、CIN组织及正常宫颈组织中Sox-2蛋白表达阳性率分别为85.71%(72/84)、36.36%(24/66)、0.18%(4/22),三者相比,P均<0.01;Sox-2 mRNA的相对表达量分别为0.877±0.208、0.463±0.093、0.243±0.023,三者相比,P均<0.01。Sox-2蛋白表达与宫颈癌淋巴结转移、临床分期、分化程度有关(P均<0.05)。Sox-2 mRNA的表达随着宫颈癌病理学分级的上升而升高。结论宫颈癌组织中Sox-2蛋白及其mRNA表达明显上调,其可能参与了宫颈癌的发生发展,并与肿瘤细胞浸润与转移有关。     

CIP2A和c-myc蛋白在宫颈癌组织中的表达及意义

目的探讨蛋白磷酸酶2A的癌性抑制因子（CIP2A）、c-myc在宫颈癌发生、发展中的作用。方法采用免疫组化SP法检测50例宫颈鳞状细胞癌、120例宫颈上皮内瘤变（CIN）组织（CINⅠ、CINⅡ、CINⅢ分别为34、40、45例）及12例正常宫颈组织中CIP2A、c-myc蛋白表达,分析二者表达与宫颈癌临床病理特征的关系及二者表达的关系。结果 CIP2A、c-myc蛋白在宫颈癌组织中的阳性表达率明显高于宫颈上皮内瘤变组织及正常宫颈组织,且与宫颈癌淋巴结转移及临床分期有关（P均〈0.05）;CIP2A与c-myc蛋白表达呈正相关（r=0.501,P〈0.05）。结论 CIP2A与c-myc在宫颈鳞状细胞癌组织中呈过表达,二者相互协调,共同促进宫颈癌的发生、发展。     

宫颈鳞癌组织中HSP70和Bcl-2的表达及意义

目的探讨热休克蛋白（HSP）70和Bcl-2在宫颈鳞癌组织中的表达及意义。方法采用免疫组化法检测HSP70、Bcl-2在60例宫颈鳞癌、18例宫颈上皮内瘤变（CIN）、7例正常宫颈组织中的表达。结果宫颈鳞癌组织中HSP70、Bcl-2阳性表达率显著高于CIN和正常宫颈组织（P均〈0．05）；宫颈鳞癌组织中HSP70的阳性表达与病理分级、临床分期、淋巴转移密切相关（P均〈0．05）；HSP70与Bcl-2在宫颈鳞癌组织中的表达密切相关（P〈0．05）。结论HSP70、Bcl-2均在宫颈鳞癌发生和发展中起重要作用，两者具有协同作用；HSP70可作为判断宫颈癌预后的重要指标。     

宫颈癌组织中COX-2的表达变化及意义

目的观察宫颈癌组织中环氧合酶（COX）-2的表达变化,并探讨其意义。方法采用免疫组化法检测45例宫颈癌、25例宫颈上皮内瘤样病变（CIN）、20例正常宫颈组织中的COX-2。结果 COX-2在正常宫颈组织中无表达,在CIN中的阳性表达率为44.0%（11/25）,在宫颈癌中为82.2%（37/45）,三者相比,P均〈0.01。G1级宫颈癌COX-2的阳性表达率高于G2、G3级（P均〈0.05）,有淋巴结转移的宫颈癌COX-2阳性表达率高于无淋巴结转移者（P〈0.05）。结论 COX-2在宫颈癌组织中过表达,可能参与了宫颈癌的发生和发展。     

宫颈癌及癌前病变组织中CD24mRNA的表达及意义

采用RT-PCR技术检测40例宫颈癌、30例宫颈癌前病变及30例正常宫颈组织中的CD24mRNA。结果显示,CD24mRNA在正常宫颈组织中不表达,在宫颈癌前病变和宫颈癌组织中的表达水平（相对值）分别为0.113&#177;0.059、0.302&#177;0.053,两者相比,P〈0.05;CD24mRNA表达与宫颈癌组织学分级有密切关系（P〈0.05）,而与临床分期无关（P〉0.05）。认为在正常宫颈到宫颈癌的演变过程中,CD24mRNA的表达逐渐呈上升趋势,可作为预测宫颈肿瘤恶变的生物学指标之一。     

MMP-2在宫颈癌中的表达及意义

采用免疫组化SP法,分别测定14例正常宫颈组织、16例宫颈原位癌、64例宫颈浸润癌组织中基质金属蛋白酶2(MMP-2)的表达.发现宫颈癌组织中MMP-2蛋白阳性率明显高于正常宫颈组织,MMP-2与临床分期、细胞分化程度及淋巴结转移均有一定关系.提示MMP-2蛋白与宫颈癌的发生、发展及转移有关,可作为观察宫颈癌进展的重要指标.     

宫颈癌组织中Fhit、survivin和Bcl-2的表达变化及意义

目的观察脆性组氨酸三联体（Fhit）、生存素（survivin）和Bcl-2在宫颈癌组织中的表达变化,并探讨其意义。方法采用免疫组化SP法检测30例正常宫颈组织、80例宫颈上皮内瘤变（CIN）组织、63例宫颈癌组织中的Fhit、survivin和Bcl-2。结果正常宫颈组织、CIN及宫颈癌组织中的Fhit失表达率分别为0、31.25%和63.49%;survivin的表达阳性率分别为0、46.25%和77.77%;Bcl-2的表达阳性率分别为6.67%、47.50%和82.54%（P均〈0.05）。Fhit表达缺失与病理学分级、临床分期和淋巴结转移无关（P均〉0.05）;survivin、Bcl-2阳性表达与宫颈癌病理学分级、临床分期和淋巴结转移有关（P均〈0.05）。结论宫颈癌组织中Fhit呈失表达、survivin、Bcl-2呈高表达,三者参与了宫颈癌发病,且survivin、Bcl-2与宫颈癌病理学分级、临床分期和淋巴结转移有关。     

IGF2BP2 and IGF2 genetic effects in diabetes and diabetic nephropathy

ObjectiveThe IGF2BP2 gene is located on chromosome 3q27.2 within a region linked to type 1 diabetes (T1D), type 2 diabetes (T2D) and diabetic nephropathy (DN). Its protein functionally binds to 5’-UTR of the imprinting IGF2 gene. The present study aims to evaluate the IGF2BP2-IGF2 genetic effects in diabetes and DN.Materials and MethodsThree cohorts including T1D with and without DN (n = 1139) of European descents from the GoKinD study, Swedish T1D with and without DN (n = 303) and Czech control subjects without diabetes, T1D, T2D with and without DN (n = 1418) were enrolled in TaqMan genotyping experiments for IGF2BP2 rs4402960 and IGF2 rs10770125. Igf2bp2 gene expression in kidney tissues of db/db and control mice at the ages of 5 and 26 weeks was examined with real time RT-PCR and Western blot.ResultsAn association of IGF2BP2 rs4402960 with T2D in the Czech population was replicated. This IGF2BP2 polymorphism (P = 0.037, OR = 0.69 95% CI 0.49–0.98) was found to be associated with DN in male not in female patients with T1D selected from the GoKinD study. In the analyses of combined the GoKinD, Czech and Swedish populations, the association between IGF2BP2 polymorphism and DN in male patients with T1D was still significant (P = 0.030, OR = 0.73, 95% CI 0.54–0.97). IGF2 rs10770125 was also associated with DN in male T1D patients of the GoKinD population (P = 0.038, OR = 0.67 95% CI 0.46–0.98). There might be a genetic interaction between IGF2BP2 and IGF2 (P = 0.05). The Igf2bp2 gene expression levels were increased in the kidneys of db/db mice compared to controls at the age of 5 weeks but not at 26 weeks.ConclusionsThe present study has replicated the association of IGF2BP2 rs4402960 with T2D in the Czech population and provided data suggesting that IGF2BP2 may have genetic interaction with IGF2 with a protective effect against DN in male patients with T1D.     

Identification of SH2B2beta as an inhibitor for SH2B1- and SH2B2alpha-promoted Janus kinase-2 activation and insulin signaling

The SH2B family has three members (SH2B1, SH2B2, and SH2B3) that contain conserved dimerization (DD), pleckstrin homology, and SH2 domains. The DD domain mediates the formation of homo- and heterodimers between members of the SH2B family. The SH2 domain of SH2B1 (previously named SH2-B) or SH2B2 (previously named APS) binds to phosphorylated tyrosines in a variety of tyrosine kinases, including Janus kinase-2 (JAK2) and the insulin receptor, thereby promoting the activation of JAK2 or the insulin receptor, respectively. JAK2 binds to various members of the cytokine receptor family, including receptors for GH and leptin, to mediate cytokine responses. In mice, SH2B1 regulates energy and glucose homeostasis by enhancing leptin and insulin sensitivity. In this work, we identify SH2B2beta as a new isoform of SH2B2 (designated as SH2B2alpha) derived from the SH2B2 gene by alternative mRNA splicing. SH2B2beta has a DD and pleckstrin homology domain but lacks a SH2 domain. SH2B2beta bound to both SH2B1 and SH2B2alpha, as demonstrated by both the interaction of glutathione S-transferase-SH2B2beta fusion protein with SH2B1 or SH2B2alpha in vitro and coimmunoprecipitation of SH2B2beta with SH2B1 or SH2B2alpha in intact cells. SH2B2beta markedly attenuated the ability of SH2B1 to promote JAK2 activation and subsequent tyrosine phosphorylation of insulin receptor substrate-1 by JAK2. SH2B2beta also significantly inhibited SH2B1- or SH2B2alpha-promoted insulin signaling, including insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1. These data suggest that SH2B2beta is an endogenous inhibitor of SH2B1 and/or SH2B2alpha, negatively regulating insulin signaling and/or JAK2-mediated cellular responses.     

解偶联蛋白2与2型糖尿病关系研究进展

2型糖尿病作为严重影响人类健康的慢性病、多发病,已经成为全球共同关注的健康及社会问题,其发病机制是目前研究的热点.最近,越来越多的研究者关注解偶联蛋白2(UCP2)与2型糖尿病的相关性,并且发现UCP2参与和影响2型糖尿病的发生、发展过程中的多个环节.     

Classification and characterization of hereditary types 2A, 2B, 2C, 2D, 2E, 2M, 2N, and 2U (unclassifiable) von Willebrand disease.

All variants of type 2 von Willebrand disease (VWD) patients, except 2N, show a defective von Willebrand factor (VWF) protein (on cross immunoelectrophoresis or multimeric analysis), decreased ratios for VWF:RCo/Ag and VWF:CB/Ag and prolonged bleeding time. The bleeding time is normal and FVIII:C levels are clearly lower than VWF:Ag in type 2N VWD. High resolution multimeric analysis of VWF in plasma demonstrates that proteolysis of VWF is increased in type 2A and 2B VWD with increased triplet structure of each visuable band (not present in types 2M and 2U), and that proteolysis of VWF is minimal in type 2C, 2D, and 2E variants that show aberrant multimeric structure of individual oligomers. VWD 2B differs from 2A by normal VWF in platelets, and increased ristocetine-induced platelet aggregation (RIPA). RIPA, which very likely reflects the VWF content of platelets, is normal in mild, decreased in moderate, and absent in severe type 2A VWD. RIPA is decreased or absent in 2M, 2U, 2C, and 2D, variable in 2E, and normal in 2N. VWD 2M is usually mild and characterized by decreased VWF:RCo and RIPA, a normal or near normal VWF multimeric pattern in a low resolution agarose gel. VWD 2A-like or unclassifiable (2U) is distinct from 2A and 2B and typically featured by low VWF:RCo and RIPA with the relative lack of high large VWF multimers. VWD type 2C is recessive and shows a characteristic multimeric pattern with a lack of high molecular weight multimers, the presence of one single-banded multimers instead of triplets caused by homozygosity or double hereozygosity for a mutation in the multimerization part of VWF gene. Autosomal dominant type 2D is rare and characterized by the lack of high molecular weight multimers and the presence of a characteristic intervening subband between individual oligimers due to mutation in the dimerization part of the VWF gene. In VWD type 2E, the large VWF multimers are missing and the pattern of the individual multimers shows only one clearly identifiable band, and there is no intervening band and no marked increase in the smallest oligomer. 2E appears to be less well defined, is usually autosomal dominant, and accounts for about one third of patients with 2A in a large cohort of VWD patients.     

CYP2J2基因和EPHX2基因多态性与缺血性脑卒中的遗传易感性

目的 研究我国汉族人群细胞色素P450表氧化酶2J2(CYP2J2),可溶性环氧化物水解酶2(EPHX2)基因多态性与缺血性脑卒中的关系.方法 选择200例缺血性脑卒中患者和350例健康人群,采用PCR-RFLP技术分析两个基因CYP2J2 G-50T,EPHX2 G860A多态性的基因型.结果 仅EPHX2 860A等位基因频率在缺血性脑卒中组与对照组比较有显著性差异.多元Logistic回归分析表明,携带EPHX2 860A等位基因的人群患缺血性脑卒中相对风险率下降50%(OR=0.5).当个体同时携带CYP2J2-50GG和EPHX2 860A (A 示A等位基因)联合基因型时,其患缺血性脑卒中相对风险率下降53.9%(OR=0.461).结论 虽然EPHX2 860A等位基因与缺血性脑卒中有相关性并且为缺血性脑卒中一个独立的保护因子,但联合基因型CYP2J2-50GG/EPHX2 860A 的协同作用对缺血性脑卒中有更强的保护作用.