hClock基因mRNA及其蛋白在结直肠肿瘤中表达的研究

目的探讨人类生物钟基因hClockmRNA及其蛋白在不同Dukes分期结直肠肿瘤中的表达,研究它们的表达与结直肠肿瘤的侵袭及转移的关系。方法采用原位杂交检测结直肠癌与相应癌旁组织中hClock基因mRNA的表达,并采用免疫组织化学检测相应标本中hCloek基因蛋白产物（CLOCK蛋白）的表达。结果21例结直肠肿瘤中hCloekmRNA弱阳性表达率47．62％,中或强阳性表达率52．38％,且与Dukes分期相关（P〈0．05）;CLOCK蛋白均呈中或强阳性表达。相应癌旁组织中hClockmRNA及蛋白呈弱阳性表达（P〈0．01）。结论hCloek基因可能与结直肠肿瘤的发生、发展及侵袭、转移有相关性。     

EMX2和β-catenin Cyclin D1在结直肠癌组织中的表达及临床意义

目的观察空通气孔同源框2(EMX2)和β-连环蛋白(β-catenin)、细胞周期蛋白D1(Cyclin D1在正常结直肠黏膜、结直肠癌组织中的表达情况,探讨结直肠癌发展过程中EMX2与β-catenin、Cyclin D1的关系。方法 S-P法检测EMX2和β-catenin、Cyclin D1在正常结直肠黏膜、结直肠癌组织中蛋白表达情况,并分析EMX2和β-catenin、Cyclin D1与结直肠癌临床病理特征之间及患者预后的关系。结果 (1)在正常结直肠黏膜、结直肠癌组织中EMX2阳性表达率分别为76. 7%和38. 2%(P 0. 01),EMX2阳性表达率与结直肠癌临床分期及淋巴结转移有关。在正常结直肠黏膜、结直肠癌组织中β-catenin阳性表达率分别为6. 7%、65. 5%; Cyclin D1阳性表达率分别为10. 0%、63. 6%。β-catenin与Cyclin D1阳性表达率与结直肠癌的临床分期、肿瘤分化程度、淋巴结转移有关(P 0. 05)。(2)在结直肠癌组织中EMX2和β-catenin表达呈负一致性(Kappa值=-0. 389,P=0. 000); EMX2与Cyclin D1表达一致性不明显(Kappa值=-0. 161,P=0. 054)。(3)应用Kaplan-Meier生存分析表明,结直肠癌组织中EMX2阳性表达与患者较好的预后有关(P=0. 032)。结论结直肠癌组织中EMX2阳性表达减少,EMX2低表达与结直肠癌患者的临床分期、淋巴结转移及预后密切相关。在结直肠癌患者中检测EMX2,可能成为判断患者临床分期和预后指标。     

Nup88在结直肠癌组织中的表达及意义

目的研究核孔蛋白复合体蛋白88（Nup88）蛋白和mRNA在结直肠癌组织中的表达及其与临床病理特征的关系。方法免疫组化方法检测181例结直肠癌标本、18例淋巴结转移癌及84例匹配的癌旁无瘤黏膜组织中Nup88蛋白表达;RT-PCR法检测Nup88 mRNA在29例结直肠癌标本及配对的癌旁2 cm组织、切缘黏膜中的表达水平。结果结直肠癌组织和淋巴结转移癌中Nup88蛋白阳性率均高于癌旁无瘤黏膜组织（P〈0.01）;Nup88蛋白的表达与结直肠癌组织分化程度有关。Nup88 mRNA在肿瘤组织中的表达水平高于癌旁无瘤黏膜和切缘黏膜（P均〈0.01）。结论 Nup88蛋白及mRNA与结直肠癌的发生发展有关,其蛋白表达可作为评估结直肠癌生物学行为的指标之一。     

Smad4表达异常与结直肠癌的相关性研究

目的 探讨结直肠组织Smad4 mRNA及其蛋白表达水平变化、Smad4基因CpG岛异常甲基化与结直肠癌及其临床病理特征的关联性.方法 应用RT-PCR方法及测序、半定量RT-PCR方法、甲基化特异性PCR(MSP)技术和免疫组织化学方法,各自检测43例患者的结直肠癌组织、癌旁组织及30例结直肠腺瘤组织、12例健康人结肠黏膜组织.结果 43例结直肠癌组织中有25例(58.14%)Smad4 mRNA检测到阳性表达,其阳性表达率(58.14%)及表达水平(0.73±0.25)均低于相应的癌旁组织(88.37%,0.95±0.29)、腺瘤组织(90.63%,1.01±0.37)和健康人黏膜组织(100.00%,1.18±0.33),差异均有统计学意义(P＜0.05).Smad4基因启动子甲基化阳性率结直肠癌组织(60.53%)明显高于癌旁组织(27.03%)、腺瘤组织(25.00%)和健康人黏膜组织(16.67%),差异均有统计学意义(P＜0.05).Smad4蛋白表达的阳性率结直肠癌组织(44.19%)明显低于癌旁组织(81.40%)、腺瘤组织(87.50%)和健康人黏膜组织(91.67%),差异均有统计学意义(P＜0.05),并随着浸润深度加深和淋巴结转移,阳性率也随之下降.结论 Smad4的低表达与结直肠癌的发展、生物学行为和预后可能有关,可作为重要的生物学标志及病期评估的标志之一.     

CKS2在结直肠癌中的表达和临床意义

背景：细胞周期蛋白依赖性激酶亚基（CKS）家族在细胞周期调节中起重要作用。研究发现其家族成员CKS2在一些恶性肿瘤中呈高表达,并与肿瘤的高侵袭性行为和预后不良相关。目前关于CKS2与结直肠癌关系的文献报道较罕见。目的：研究CKS2在结直肠癌中的表达和临床意义。方法：应用realtimeliT—PCR和蛋白质印迹法检测23例临床结直肠癌手术标本癌组织、癌旁非癌组织和正常组织中的CKS2mRNA和蛋白表达。结果：CKS2mRNA和蛋白在结直肠组织中的相对表达量依次为：癌组织〉癌旁组织〉正常组织,癌组织与正常组织问差异有统计学意义（P〈0．01）。性别对CKS2mRNA在不同结直肠组织中的表达趋势无明显影响。癌组织中的CKS2蛋白表达与肿瘤大小和pTNM分期呈正相关（P〈0．01）,与肿瘤部位无关。结论：CKS2蛋白在结直肠癌中呈高表达并与肿瘤临床病理特征相关,有望成为结直肠癌新的分子标记物和治疗靶点。     

结直肠侧向发育型肿瘤组织中RNF6蛋白的表达变化及其意义

目的 观察结直肠侧向发育型肿瘤（LST）组织中RNF6蛋白的表达变化，并探讨其意义。方法 LST患者33例、结直肠隆起型腺瘤（PA）患者37例、结直肠癌患者35例均接受手术治疗，手术过程中留取切除的肿瘤组织分别记为LST组、PA组、CRC组，留取结直肠癌患者的癌旁正常组织（距癌组织>5 cm）记为Control组。采用免疫组织化学染色法检测各组织中RNF6蛋白，采用半定量评分法评估蛋白表达情况，包括阳性表达率和阳性程度，并分析RNF6蛋白表达与LST患者临床病理参数的关系。结果 癌旁正常组织多数不染色或染色颜色较浅、阳性面积小，而结直肠癌组织中RNF6蛋白染色较深，多数呈棕黄色颗粒沉积，PA组织及LST组织的染色程度介于癌旁正常组织及结直肠癌组织中间。Control组、PA组、LST组、CRC组RNF6蛋白阳性表达率分别为40%、51.4%、75.8%、91.4%，组间相比，P均<0.05。Control组、PA组、LST组、CRC组RNF6蛋白阳性强度呈递增趋势（P均<0.05）。RNF6蛋白表达与LST患者上皮内瘤变级别有关（P<0.05），上皮内瘤变级别越...     

MTSS1及Gli1在食管鳞癌组织中表达及其相关性

目的 检测食管鳞癌组织中肿瘤转移抑制蛋白(MTSS)1及胶质瘤相关癌基因蛋白(Gli)1的表达水平,探讨MTSS1和Gli1的相关性。方法 应用免疫组织化学SP法和原位杂交方法检测136例食管鳞癌组织及其相应的96例癌旁不典型增生组织和136例正常食管黏膜组织中MTSS1、Gli1蛋白及mRNA的表达水平。结果 MTSS1蛋白和mRNA在食管鳞癌组织中的阳性表达率显著低于癌旁组织和正常食管黏膜组织,两两比较差异均有统计学意义(P<0.05)。Gli1蛋白和mRNA在食管癌组织中的阳性表达率显著高于癌旁不典型增生组织和正常食管黏膜组织,两两比较差异均有统计学意义(P<0.05)。有淋巴结转移组食管鳞癌组织中MTSS1蛋白及mRNA阳性表达率均显著低于无淋巴结转移组(P<0.05);有淋巴结转移组食管鳞癌组织中Gli1蛋白及mRNA阳性表达率均显著高于无淋巴结转移组(P<0.05);Gli1和MTSS1蛋白在食管鳞状上皮细胞癌组织中的表达呈负向相关(r=-0.422,P<0.05),Gli1和MTSS1 mRNA在食管鳞状上皮细胞癌组织中的表达亦呈负向相关(r...     

结直肠癌组织COX-2 mRNA表达的临床病理意义

目的检测环氧合酶-2(COX-2)mRNA在结直肠癌、癌旁及正常组织中的表达情况.探讨其表达与临床病理特征的关系.方法应用逆转录聚合酶链反应(RT-PCR)技术检测24例结直肠癌、癌旁和正常组织中COX-2的mRNA表达.结果 24例结直肠癌中,COX-2 mRNA表达阳性者17例,癌旁和正常组织阳性者分别为9例和3例,癌组织中的表达率明显高于癌旁和正常组织(P＜0.01);COX-2 mRNA的阳性表达与结直肠癌的淋巴结转移、远处转移、Duke′s分期呈正相关.结论结直肠癌组织中COX-2 mRNA表达水平高于癌旁和正常组织,其表达与结直肠癌侵袭转移密切相关.因此COX-2mRNA可作为预测结直肠癌细胞转移潜能的敏感指标.     

APPL1表达与结直肠癌发生发展的关系及意义

目的:研究APPL1蛋白在人结直肠癌组织中的表达情况与临床病理参数的关系.方法:收集35例新鲜结直肠癌及27例正常直肠黏膜组织,采用免疫组织化学SP法和RT-PCR法检测APPL1在结直肠癌组织及正常直肠黏膜组织中的表达.采用半定量积分分级对该蛋白的表达强弱进行评分.结果:免疫组织化学和RT-PCR结果显示,APPL1蛋白在结直肠癌组织以及正常直肠黏膜组织中普遍表达,但该蛋白和相应的mRNA在癌组织中的表达高于对照组(P<0.05).在35例结直肠癌组织中,APPL1表达与分化程度、淋巴结转移、TNM分期相关(P<0.05),与性别、年龄、肿瘤大小、组织学类型无明显相关(P>0.05).结论:APPL1蛋白在结直肠癌组织中的表达上调,且该蛋白表达与患者肿瘤的分化程度、淋巴结转移情况以及TNM分期有关.APPL1有可能成为结直肠癌治疗的一个新靶点.     

Kiss-1在食管鳞状细胞癌组织中的表达及临床病理意义

目的:观察Kiss-1蛋白及mRNA在食管鳞状细胞癌组织中的表达及其临床病理意义.方法:采用免疫组织化学SP法、原位杂交和逆转录聚合酶链反应(RT-PCR)技术,联合检测了62例食管鳞状细胞癌、31例癌旁不典型增生及62例正常食管组织中Kiss-1蛋白和mRNA的表达,并探讨其临床病理意义.结果:在食管鳞状细胞癌组织、癌旁不典型增生及正常食管组织中,Kiss-1蛋白的阳性表达率分别为56.5%、67.7%、90.3%(P<0.05);用原位杂交检测Kiss-1 mRNA阳性表达率分别为51.6%、74.2%、95.2%(P<0.05);用RT-PCR技术检测Kiss-1 mRNA阳性表达率分别为54.8%、71.0%、88.7%(P<0.05);食管鳞状细胞癌组织中Kiss-1蛋白及mRNA的低表达与淋巴结转移密切相关(P<0.05),而与患者的性别、年龄、肿瘤的组织学分级无关(P>0.05);Kiss-1蛋白的低表达与浸润深度有关(P<0.05);在食管鳞状细胞癌组织中,Kiss-1蛋白及mRNA的表达呈正相关(P<0.05),用原位杂交及RT-PCR技术对Kiss-1 mRNA表达情况的检测结果也呈正相关(P<0.05).结论:Kiss-1基因的表达降低或缺失和食管鳞状细胞癌的发生发展及转移有关,有望成为食管鳞状细胞癌早期诊断和预后判断的重要指标之一.     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Study of constancy of hydrogen-consuming flora of human colon

The constancy of the hydrogen consuming flora of the human colon was studied in 15 healthy subjects via two measurements obtained 18 to 36 months apart. Hydrogen disappearance rate and the major products of H₂-consuming bacteria, methane and sulfide, were measured during incubation of fecal homogenates with excess hydrogen and sulfate. In 11/15, the hydrogen consumption rate and the predominant hydrogen-consuming pathway (methanogenesis, sulfate reduction, or neither) remained constant. However, major shifts in these pathways were observed in four subjects, with two losing and two gaining the ability to produce methane. Methanogenesis was associated with the highest hydrogen consumption rate. This study demonstrates that clinically unrecognizable, major alterations of the colonic flora occur in healthy subjects. Understanding of the factors responsible for these alterations might allow for therapeutic manipulation of the colonic flora.Supported in part by the Department of Veterans Affairs and NIDDKD RO1 DK 13309-25.