胃癌中Survivin、Cox-2与HIF-1α表达的相互关系及临床意义

目的:探讨胃癌组织中Survivin、Cox-2与HIF-1α蛋白的表达及其临床意义。方法:采用免疫组织化学染色法检测Survivin、Cox-2及HIF-1α在65例胃癌组织、18例癌旁组织及20例正常胃黏膜的表达水平,并分析其与胃癌生物学行为的关系。结果:Survivin、Cox-2与HIF-1α在正常胃黏膜、癌旁组织、癌组织中的表达呈升高趋势,其差异均有统计学意义(P0.05)。Survivin阳性表达在TNM分期、组织学分化程度、胃癌浸润深度、淋巴结转移及远隔转移方面差异显著(P0.05);Cox-2阳性表达在TNM分期、组织学分化程度和淋巴结转移方面差异显著(P0.05);HIF-1α阳性表达在TNM分期、组织学分化程度、胃癌浸润深度和淋巴结转移方面差异显著(P0.05)。相关分析显示,65例胃癌组织中,Survivin、Cox-2与HIF-1α表达均呈正相关(r=0.285,0.405,0.546,P0.05)。结论:Survivin、Cox-2与HIF-1α的过表达在胃癌的发生、发展、浸润和转移过程中发挥协同作用,可成为胃癌基因诊断和治疗的新靶点。     

胃癌组织中miR-107、AXIN2的表达及与预后的相关性

目的检测微小RNA-107(microRNA-107, miR-107)和轴抑制蛋白2(axis inhibition protein 2, AXIN2)在胃癌组织中的表达,探讨两者表达的关系及临床意义。方法采用qRT-PCR法检测65例胃癌及正常胃黏膜组织中miR-107和AXIN2 mRNA表达水平;采用免疫组化SP法检测AXIN2蛋白表达水平,分析miR-107、AXIN2与胃癌临床病理特征的关系;Kaplan-Meier法分析胃癌患者术后3年生存期;Cox生存回归分析影响胃癌患者预后的危险因素;Pearson分析胃癌组织中miR-107与AXIN2 mRNA表达水平的相关性。结果胃癌组织中miR-107的表达水平高于正常胃黏膜组织(P0.05),AXIN2 mRNA表达水平及蛋白阳性率均低于正常胃黏膜组织(P0.05);miR-107和AXIN2表达与肿瘤分化程度、淋巴结转移、TNM分期及浸润深度有关(P0.05);miR-107高表达组患者的3年生存率显著低于miR-107低表达组患者,AXIN2高表达组胃癌患者3年生存率显著高于低表达组患者(P0.05);淋巴结转移、TNM分期、miR-107高表达、AXIN2低表达是影响胃癌患者不良预后的独立危险因素(P0.05);胃癌组织中miR-107与AXIN2 mRNA表达呈负相关(r=-0.607,P0.001)。结论胃癌组织中miR-107高表达、AXIN2低表达,且与患者预后有关,可能作为胃癌患者潜在的预后标志物。     

胃癌组织中CXCR3表达与微血管形成的关系及其临床意义

目的探讨胃癌组织中CXCR3的表达和微血管形成的关系并分析其临床意义。方法采用免疫组化法检测169例胃癌组织中CXCR3蛋白表达和肿瘤间质微血管密度(microvascular density,MVD),分析其相关性及其与胃癌临床病理参数的关系。结果 CXCR3表达在分化程度高(P=0.025)、浸润程度浅(P=0.005)、TNM分期低(P=0.002)及无淋巴结转移(P=0.001)的胃癌患者中更高;MVD在胃癌分化程度低者中更高(P=0.001),与浸润深度、TNM分期及淋巴结转移无关(P0.05)。胃癌组织中CXCR3表达与MVD呈负相关(r=-0.151,P=0.049)。Kaplan-Meier分析发现高表达CXCR3的胃癌患者(P=0.015)和低MVD患者(P=0.047)的生存时间延长。结论胃癌中CXCR3的表达与MVD呈负相关,且与胃癌的浸润、转移相关。     

MARCH2在胃癌中的异常表达及临床意义

目的检测胃癌组织中MARCH2蛋白表达并分析其与临床病理特征的相关性。方法采用免疫组化SP法检测125例胃癌组织和30例正常胃黏膜组织中MARCH2蛋白表达,并复习相关文献。结果 MARCH2在125例胃癌组织中的阳性率为72.8%,在正常胃黏膜组织中的阳性率为23.3%。MARCH2表达与胃癌分化程度、浸润深度、淋巴结转移和TNM分期密切相关。结论 MARCH2在胃癌组织中高表达,与胃癌分化程度、深部浸润、淋巴结转移、更高的TNM分期密切相关。MARCH2可能成为胃癌诊断、治疗和预后的分子靶标。     

整合素αvβ6和MMP-9在胃癌中表达及其与胃癌病理生物学特征的关系

目的:探讨整合素αvβ6、MMP-9在胃癌中表达及其与胃癌病理生物学特征的关系。方法:采用EnVision免疫组化方法检测94例胃癌及癌旁胃黏膜组织中整合素αvβ6、MMP-9的表达情况,并分析两者表达与胃癌病理生物学特征的关系。结果:胃癌组织中整合素αvβ6和MMP-9阳性表达率明显高于癌旁胃黏膜组织(P<0.01)。整合素αvβ6在胃癌组织中的表达与Lauren分型、组织分化程度、是否有淋巴结转移(N分期)以及TNM分期密切相关(P<0.01),而MMP-9在胃癌组织中的表达与Lauren分型、组织分化程度、是否有淋巴结转移(N分期)、浸润深度以及TNM分期密切相关(P<0.01)。在胃癌组织中整合素αvβ6和MMP-9的表达呈正相关(r=0.672,P<0.01)。整合素αvβ6和MMP-9表达阴性患者的5年生存率明显优于整合素αvβ6和MMP-9表达阳性的患者(P<0.01)。Cox回归多因素分析表明Lauren分型、是否有淋巴结转移(N分期)、整合素αvβ6和MMP-9高表达是影响患者5年生存率的独立预后因素(P<0.05)。结论:整合素αvβ6和MMP-9的高表达与胃癌的浸润、转移密切相关;两者表达可作为判断胃癌患者预后的参考指标及治疗的靶向目标。     

胃癌组织中CapG蛋白的表达及其与临床病理特征及预后的关系

目的检测巨噬细胞加帽蛋白(gelsolin-like actin-capping protein,CapG)在胃癌及癌旁组织(距离癌组织边缘5 cm)中的表达,探讨CapG蛋白表达与胃癌临床病理参数及预后的关系。方法收集125例胃癌石蜡包埋组织样本及癌旁组织30例。患者均为未行放、化疗的初诊患者。每例组织样本均行CapG免疫组化染色,并设置对照组。通过病理医师评分来确定CapG蛋白的表达量,探讨CapG蛋白表达与胃癌临床病理参数及预后的关系。结果 CapG蛋白在细胞质和细胞核中均表达,呈棕黄色颗粒状。癌旁组织中基本无表达,胃癌组织中的表达显著高于癌旁组织;胃癌组织中CapG蛋白表达与患者性别、年龄、肿瘤大小、肿瘤位置无显著相关(P均0.05),与浸润深度(P=0.044)、淋巴结转移(P=0.026)、远处转移(P=0.001)及AJCC分期(P=0.012)显著相关。Kaplan-Meier生存曲线显示,CapG蛋白高表达组患者的总体预后较低表达组差,且差异具有显著性(P0.001)。Cox回归分析显示,肿瘤位置、淋巴结转移、远处转移、AJCC分期、CapG蛋白表达情况是胃癌患者预后的独立危险因素。结论 CapG蛋白在胃癌组织中高表达,可能与胃癌的发生、发展有关,可作为胃癌预后判断的指标之一,有望成为胃癌治疗的潜在新靶点。     

胃癌中CMTM6和PD-L1的表达及其临床意义

目的 探讨CMTM6和PD-L1在胃癌中的表达及其与临床病理特征、预后的关系。方法 收集103例胃癌患者的临床资料，采用免疫组化MaxVision两步法检测胃癌组织中CMTM6和PD-L1的表达，分析两者表达与临床病理特征的关系。结果 103例胃癌组织中CMTM6高表达70例(68.0%),低表达33例(32.0%);PD-L1高表达66例(64.1%),低表达37例(35.9%)。正常胃黏膜组织中CMTM6高表达39例(37.9%)、低表达64例(62.1%);PD-L1高表达35例(34.0%)、低表达68例(66.0%);两组相比差异有显著性(P<0.001)。CMTM6表达与肿瘤分化程度、浸润深度、脉管浸润、淋巴结转移、远处转移、TNM分期和PD-L1表达相关(P<0.05)。PD-L1表达与患者性别、脉管浸润、神经侵犯和TNM分期相关(P<0.05)。Kaplan-Meier生存分析表明，CMTM6高表达与胃癌患者不良预后相关(P=0.008)。Cox单因素生存分析显示，胃癌患者的预后与肿瘤最大径、浸润深度、脉管浸润、远处转移、淋巴结转移、TNM分期和CMT...     

肺癌中Eg5、Ki-67的表达及临床意义

目的观察Eg5、Ki-67在肺癌中的表达及与临床病理特征的关系,探讨两者与肺癌预后的关系。方法采用免疫组化法检测79例肺癌组织、20例肺良性病变组织中Eg5、Ki-67蛋白表达,分析两者表达与临床病理特征的关系;并随访患者生存期,采用Kaplan-Maier生存分析和Cox回归法分析Eg5、Ki-67在肺癌组织中表达和预后的关系。结果 Eg5在肺癌组织中的阳性率为65.8%,Ki-67在肺癌组织中的强阳性率为48.1%,Eg5、Ki-67表达均与患者临床分期、淋巴结转移明显相关。Kaplan-Maier生存分析提示Eg5阳性、淋巴结转移和中晚期的肺癌患者预后较差(P0.05);Cox多因素回归分析显示临床分期、淋巴结转移是影响肺癌患者预后的独立危险因素,Eg5表达与淋巴结转移明显相关(r=0.539,P0.05)。结论 Eg5蛋白在肺癌组织中呈阳性表达;Eg5、Ki-67蛋白高表达与淋巴结转移密切相关,Eg5阳性提示肺癌患者预后不良。     

COX-2与P73在胃癌中的表达及相关性探讨

目的：探讨COX-2和P73在胃癌组织中的表达及临床意义。方法：应用免疫组化Envision法检测COX-2和P73在62例胃癌组织中的表达情况。结果：全组COX-2阳性率为64.5%（40/62），P73阳性率为69.4%（43/62）。COX-2的表达与组织分化程度、浸润深度、淋巴结转移和临床TNM分期相关（P〈0.05）；P73的表达与浸润深度、淋巴结转移和临床TNM分期相关（P〈0.05）。COX-2和P73同时表达、单独表达及不表达时，胃癌的组织分化程度、病变浸润深度（T分期）、淋巴结转移（N分期）和临床分期组间差异有统计学意义（P〈0.05）。COX-2和P73在胃癌组织中的表达具有相关性（r=0.384，P〈0.05）。结论：胃癌组织中存在COX-2和P73基因的过度表达，COX-2和P73基因的过度表达与胃癌发生发展有关。     

Ki-67在胃癌组织浸润和转移中的作用

目的探讨Ki-67在胃癌组织浸润和转移中的表达和意义。方法采用SP免疫组化法检测64例胃癌组织中Ki-67基因表达情况。结果 Ki-67表达在淋巴结转移组高于无淋巴结转移组(P<0.01),在浸润组的表达高于无浸润组的表达(P<0.01),Ki-67在低分化组的表达高于中高分化组(P<0.01)。结论 Ki-67抗原表达的检测对评估胃癌的浸润与否,淋巴结转移与否以及胃癌分化程度有重要的意义,可帮助估计胃癌预后。     

RAS蛋白在胶质瘤中的表达及其对胶质瘤细胞生长的影响

目的 探讨RAS蛋白在胶质瘤中的表达及其对人脑胶质瘤细胞生长的影响.方法 用免疫组化染色法检测RAS蛋白在正常脑组织及各级别胶质瘤组织中的表达水平,并采用四甲基偶氮唑盐微量酶反应比色法和流式细胞技术检测降低RAS活性后U251细胞的增殖和凋亡情况;用Western印迹法检测ERK和AKT信号通路.结果 胶质瘤组织中RAS的表达随胶质瘤恶性程度增高而增强.抑制RAS 蛋白活性后,RAS信号通路的下游分子ERK和AKT蛋白磷酸化水平降低;U251胶质瘤细胞生长受抑制,细胞生长阻滞在G1期且凋亡增加.结论 RAS蛋白在胶质瘤中高表达,抑制其活性可下调ERK和AKT 信号通路,进而调控细胞的生长.     

Isolation of human C-reactive protein and serum amyloid P component

Procedures are described for the isolation in high yield of consistent, highly purified preparations of human C-reactive protein (CRP) and serum amyloid P component (SAP). CRP was obtained from malignant ascitic and pleural fluids by calcium-dependent affinity chromatography on pneumococcal C-polysaccharide covalently coupled to cyanogen bromide-activated Sepharose. It was then gel filtered on Ultrogel AcA44 (acrylamideagarose beads) in the presence of calcium ions, combining molecular sieve chromatography with removal of contaminating SAP by its affinity for agarose. Residual trace contaminants were removed by immunoabsorption with anti-normal human serum and anti-SAP antibodies insolubilised on Sepharose and/or by absorption with Sepharose-Con A to remove glycoproteins and Blue-Sepharose to remove albumin. After a final gel filtration step on Sephacryl S-300 35–44% of the initial CRP was recovered in pure from according to biochemical and immunochemical criteria. SAP was isolated from normal serum by calcium-dependent affinity chromatography on unsubstituted Sepharose beads, followed by solid-phase immunoabsorption of contaminants and finally gel filtration on Sephacryl S-300. At least 50% of the SAP in the starting material was recovered in pure form according to biochemical and immuunochemical criteria. Ready availability of such preparations facilitates biochemical, biophysical and particularly biological studies of these plasma proteins.     

Binding of mannan-binding protein to various bacterial pathogens of meningitis

Mannan-binding protein (MBP), a calcium-dependent plasma lectin, may play a role in the innate defence against microorganisms. After binding lo carbohydrate structures at the bacterial surface, MBP activates the classical pathway of the complement system. To investigate the binding capacity of MBP to various bacteria associated with meningitis, an assay was developed to study the binding of MBP to bacteria grown in a semisynthetic fluid culture medium. Salmonella montevideo (containing a mannose-rich lipopolysaccharide (EPS)), used as a positive control strain, showed binding of radiolabelled MBP at a level of 80% compared with binding of MBP to zymosan. Binding of labelled MBP to Salm. montevideo was time-dependent, temperature-dependent and saturable. The binding, was inhibited by unlabelled MBP., by mannose and by N-acetyl-o-glucosamine. Among bacterial pathogens often found to cause meningitis, a wide range of MBP binding capacities could be determined. The encapsulated Neisseria meningitidis (representatives from 11 serogroups other than group A were included: n = 22), N. mucosa (n = 1), Haemophilus influenzae type b (n = 10) and Streptococcus agalactiae (n = 5) had a low MBP binding capacity of 21.7% (95% confidence interval (Cl) 3.3–40.1%). Escherichia coli K1 (n =11). Strep, suis (n = 5), Strep, pneumoniae (n = 10) and N. meningitidis scrogroup A (n = 2) showed intermediate MBP binding capacity of 58.4% (95% Cl 40.0–76.8%). A third group consisting of non-encapsulated Listeria monocytogenes (n = 11), non-encapsulated H. influenzae (n = 2), non-encapsulated N. meningitidis (n = 2), N. cinera (n = 1) and N. subflava (n = 1) strains had a high MBP binding capacity of 87.5% (95% CI 62.5–12.5%). The majority of encapsulated pathogens causing bacterial meningitis seem to have a rather low MBP binding capacity.     

β淀粉样蛋白对大鼠学习记忆功能及tau蛋白异常磷酸化的影响

为了探讨β淀粉样蛋白对大鼠学习记忆功能和tau蛋白异常磷酸化的影响,本文在海马注射Aβ25-35建立阿尔茨海黙病(AD)大鼠模型的基础上,通过行为学检测、HE染色、免疫组化和免疫蛋白印迹技术对动物的学习能力、组织的病理改变和tau(pS202)、tau(pT231)和tau-5的表达情况进行了分析。在行为学检测中,Aβ注射组大鼠在穿梭箱实验中的主动回避次数和被动回避次数减少,失败次数增多,而在Morris水迷宫测试中的逃避潜伏期和游泳距离延长。HE染色显示Aβ注射组大鼠海马CA1、CA3、齿状回的神经细胞数目减少;而免疫组化和免疫印迹结果显示注射组tau(pS202)阳性细胞明显增加,tau(pS202)、tau(pT231)和tau-5蛋白表达增加。以上结果提示海马内注射Aβ25-35可引起大鼠学习记忆功能下降,可能与神经细胞减少,tau蛋白异常磷酸化增多有关。     

The concentration of the C-type lectin, mannan-binding protein, in human plasma increases during an acute phase response.

Two ELISAs for estimating mannan-binding protein (MBP) were constructed and the concentration of MBP in plasma was followed in patients undergoing major surgery and in patients having a malarial attack. In both cases increases of MBP in the plasma were observed. The relative increase and the kinetics varied from person to person. The concentration of MBP increased between 1.5- and three-fold following surgery. In some patients an increase was seen at day 1 whereas in others the increase was not observed until days 3-9. In the malaria patients an increased level of MBP was maintained during 30 days of treatment with chloroquine. The relative increase in MBP was independent of the presurgery or premalaria levels.     

La蛋白质家族与肿瘤关系

La蛋白相关家族(La-related proteins,LARPs)包括LARP1、LARP1b、LARP3,(Genuine La)、LARP4a、LARP4b、LARP6和LARP7.LARPs结构包含中度保守的RNA识别模体(RNA recognition motif,RRM)和多个其他结构、运输元件,可定位于不同的亚细胞部位与RNA相互作用,在细胞转录和翻译等方面发挥重要的作用.人LARPs最初是在系统性红斑狼疮和Sjogren's综合征患者的血清内发现的自体抗原,目前随着研究深入普遍认为LARPs作为一种RNA分子伴侣参与RNA代谢能发挥多种生物学功能.此前国内有学者研究验证LARPs能稳定乙型肝炎病毒(hepatitis B virus,HBV)的mRNA,帮助其在肝细胞中复制与翻译,在乙型肝炎进展中具有重要作用.近年发现,LARPs在多种恶性肿瘤如子宫颈癌、乳腺癌、肝癌和前列腺癌等的细胞增殖、分化、迁移、血管形成的调节等方面起重要作用.而恶性肿瘤的发生与演进是由环境与遗传因素相互作用而导致的许多基因突变,实际上基因突变产生的编码蛋白质执行细胞生命活动,因而LARPs在肿瘤中扮演的分子角色将可能在肿瘤的诊断与治疗中提供潜在的新靶点、新策略和新途径.     

Role of a conserved sequence in the maturation and function of the NDV HN glycoprotein

The hemagglutinin-neuraminidase (HN) proteins of viruses in the Paramyxouirus genus have a short conserved sequence, G(A, S)EGR(I, L, V). The role of this sequence in the intracellular processing and function of the Newcastle disease virus HN protein was explored by site directed mutagenesis. Mutations in this region fall into two categories. One set of mutants (G398A, E400D, R402K, and a deletion removing amino acids 400–403) was defective in folding. These mutant proteins formed little or no mature, disulfide linked oligomer. They had few or no antigenic sites found on the mature protein and they were transported to the cell surface poorly or not at all. The second class of mutants (A399G, G401A, G401L) was minimally affected in folding and intracellular transport. When normalized to surface expression, this group of mutant proteins had wild type levels of attachment activity, neuraminidase activity, and fusion promotion activity. Thus mutations in this region directly affect intracellular processing but not the biological activities of the protein. This sequence may, therefore, be conserved in the HN proteins of Paramyxoviruses because it is critical to the folding of the molecule.     

上皮钙黏蛋白、波形蛋白和Snail蛋白表型在乳腺浸润性导管癌中表达的相关性及其与预后的关系

目的 检测上皮钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)和Snail蛋白在乳腺浸润性导管癌中的表达情况与乳腺癌病理特征的关系及其与浸润性导管癌预后的关系.方法 采用Envision免疫组织化学方法对89例乳腺浸润性导管癌组织中的上E-cadhenn,Vimentin和Snail蛋白表达情况进行检测并分析表达程度与临床病理特征之间的关系;统计分析采用卡方检验、Fisher精确概率法检验Spearman等级相关检验、配对计数资料检验、Kaplan-Meier法进行单因素生存分析,Cox比例风险模型进行生存的多因素分析.结果 E-cadherin、Vimentin和Snail蛋白在浸润性导管癌组织中的阳性表达率分别为、47.1％、53.9％和55.0％,三者呈负相关,(P=0.003);E-cadherin、Vimentin和Snail蛋白的表达与临床TNM分期有关(P ＜0.005).浸润性导管癌中Snail蛋白、Vimentin表达明显增高,Snail蛋白与淋巴结转移有关(P =0.029);Vimentin与淋巴结转移、雌激素状态有关(P =0.006,P＜0.001);E-cadherin表达明显降低,与孕激素状态有关(P =0.030);分子分型结果显示,管腔A型、HER-2阳性型组的Vimentin、Snail蛋白阳性表达率低于基地细胞样型组(P =0.012),而E-cadherin表达率则高于基地细胞样型组(P =0.004).Cox分析发现,E-cadherin低表达和Vimentin的过度表达与患者总生存期(P =0.019、P=0.045)及患者无病生存期(P =0.032、P=0.024)显著相关,但Snail蛋白的表达与总生存期及无病生存期无相关性(P =0.879、P=0.835).结论 E-cadhern、Vimentin和Snail蛋白在乳腺浸润性导管癌的发生、发展、侵袭及转移中起重要作用,对认识乳腺癌的生物学特性以及对指导乳腺癌的诊疗及预后具有重要意义.     

Characterization of newly identified DnaA and DnaB proteins from Acetobacter

Although chromosomal replication is an essential feature of the bacterial life cycle, the replication mechanism and involved molecular players have never been properly characterized in the Acetobacter genera. Thanks to whole-genome sequencing, the unknown replication proteins from Acetobacter pasteurianus and Acetobacter orleanensis, DnaA-like and DnaB-like, could be identified. Despite the low nucleotide or amino acid similarity to the respective orthologs from Escherichia coli, their involvement during replication regulation was corroborated by artificial microRNA. In the Acetobacter genome, a novel replication origin, oriAo, was detected with three 9-nucleotide-long DnaA boxes to which DnaA-like proteins bind actively. Bacterial two-hybrid systems and co-immunoprecipitation confirmed the homologous and heterologous interactions between DnaA-like and DnaB-like proteins with their E. coli orthologs. This communication is due to the conserved tryptophan at position 6 for E. coli or 25 for Acetobacter that unables DnaA-like proteins to form oligomeric protein structures after its substitution. Altogether, these results provide novel insights into the genome replication mechanism in Acetobacter.