瘦素对SD乳鼠骨胳肌细胞葡萄糖氧化代谢的影响

瘦素是肥胖基因的表达产物 ,由脂肪细胞分泌 ,对糖、脂代谢产生影响 ,是机体维持体重和能量代谢平衡不可缺少的激素 ,但它对骨胳肌葡萄糖代谢以及对胰岛素作用的影响 ,众多研究结果报道不一。本研究旨在初步探讨瘦素对骨胳肌细胞葡萄糖氧化的影响 ,并观察其剂量效应以及生理剂量瘦素对胰岛素作用的影响。一、材料和方法1.材料 :正常雄性ＳＤ乳鼠 (两天龄 ) 3只 ,购自中山医科大学动物实验中心 ;Ⅲ型胶原酶购自Ｓｉｇｍａ公司 ;胎牛血清、ＤＭＥＭ、Ｆ12购自Ｈｙｃｌｏｎｅ公司 ;Ｄ 〔Ｕ 14 Ｃ〕葡萄糖 (放射性比活性 9.2× 10 3 ＭＢｑ/ｍ…     

缺氧对C57BL/6J小鼠瘦素及其受体基因表达的影响

目的　研究缺氧对小鼠瘦素及其受体 (包括转运性受体Ra、Rc和功能性受体Rb)mRNA表达变化的影响,以进一步明确瘦素和呼吸功能的关系。方法　利用医学自动测控缺氧仓(XQ Ⅰ型)复制机体常压缺氧状态,将小鼠分为正常对照组、缺氧 24h组、缺氧 48h组,然后利用逆转录 聚合酶链反应(RT-PCR)分别定量检测瘦素及其受体mRNA表达水平。根据cDNA条带的灰度值计算其相对含量,各种产物与磷酸甘油醛脱氢酶 (GADPH)的比值表示其相对表达水平。结果(1)缺氧 24h组和缺氧 48h组瘦素mRNA表达水平分别为 0.903±0.190、0.856±0.336,显著高于正常对照组(0.508±0.207,P均<0.05); (2)缺氧 24h组和缺氧 48h组RamRNA表达水平分别为0 724、0.700,分别为正常对照组 (0 630)的 115%、111%; (3)缺氧 24h组和缺氧 48h组RbmRNA表达水平分别为 0.381±0.038、0.345±0.042,与正常对照组(0.258±0.049)比较差异均有统计学意义(P均<0.05); (4)缺氧 24h组和缺氧 48h组RcmRNA表达水平分别为 0.299、0.292,分别为正常对照组(0.133)表达水平的 224%、219%。结论　缺氧作为一种独立因素可在一定范围内刺激机体瘦素、瘦素受体Ra、Rb、Rc基因表达增加,从而在正向调节机体呼吸功能中发挥重要作用。     

脂肪组织瘦素基因与非酒精性脂肪肝的研究

目的探讨非酒精性脂肪肝(NAFLD)患者脂肪组织瘦素mRNA基因表达水平和胰岛素抵抗与血浆瘦素等的相关性。方法行熳性胆囊炎、胃溃疡、腹股沟疝等择期手术的NAFLD患者21例、对照组患者24例,于术中取少许腹部皮下和网膜脂肪组织送检。应用SYBR Green I实时定量逆转录聚台酶链反应法检测瘦素mRNA的表达水平,用稳态模型法计算胰岛素抵抗指数,用酶联免疫吸附法测定血浆瘦素和胰岛素水平。结果NAFLD和对照组瘦素基因表达值分别为1．32±0．12、0．99±0．05,1．10±0．09、0．87±0．13；瘦素基因表达和胰岛素抵抗指数与血浆瘦素浓度直接关联(r值分别为0．72、0．69,P值均＜0．05)。结论脂肪组织瘦素基因高表达是高瘦素血症的主要原因,肥胖和非肥胖的NAFLD患者存在瘦素抵抗和胰岛素抵抗,提示瘦素抵抗与胰岛素抵抗一样和NAFLD发病密切相关。     

抵抗素基因转染诱导肝性胰岛素抵抗细胞的鉴定

目的 将编码人抵抗素基因的pcDNA 3.1(+)真核表达载体在HepG2肝癌细胞中进行稳定转染,以建立抵抗素诱导的肝性胰岛素抵抗细胞模型.方法 实验分3组:HepG2细胞组;空质粒组;抵抗素基因转染组,通过免疫细胞化学和RT-PCR方法 进行抵抗素基因和蛋白表达鉴定,用微量化的葡萄糖氧化酶法检测细胞对葡萄糖的摄取作用.结果 免疫细胞化学染色结果 表明:与对照组比较,抵抗素基因转染组抵抗素蛋白平均光密度值显著升高(P＜0.01);而空质粒组无显著差异(P＞0.05).RT-PCR扩增产物电泳表明:抵抗素基因转染组有目的 条带出现,而对照组和空质粒组无目的 条带出现,细胞葡萄糖摄取实验表明:抵抗素基因转染细胞对葡萄糖摄取作用下降.结论 过度表达人抵抗素基因的胰岛素抵抗的肝细胞建模成功.     

RNA干扰对大鼠血管内皮细胞血管紧张素转换酶表达的抑制作用

目的用RNA干扰技术选择性下调大鼠血管内皮细胞(ECs)上血管紧张素转换酶(ACE)的表达。方法构建携带ACE基因特异短发夹RNA(shRNA)编码序列的质粒载体pACE-shRNA,与脂质体转染剂制备成不同比例的复合物,转染原代培养的大鼠主动脉ECs,在荧光显微镜下检测转染效率,用四甲基氮唑盐比色法检测细胞存活率,选择出质粒与转染剂的最佳配比。然后以最佳配比复合物转染ECs,分别于转染前及转染后24、48、72h通过半定量RT-PCR和Western blot法检测ACE的表达。结果质粒与转染剂的最佳配比为质粒1.0μg、转染剂4μl,此时转染效率为(81.2±6.2)%,细胞生存率为(90.2±4.1)%。以最佳配比复合物转染ECs后24h,ACE的mRNA和蛋白质表达无明显变化;转染后48h,表达明显降低,差异有统计学意义(P<0.01);72h时表达更低,只有微弱的条带显示。结论RNA干扰能选择性下调原代培养的大鼠血管ECs上ACE的表达,这可能为心血管疾病基因治疗的研究和应用提供新方法。     

HBV影响TRIM22表达的机制及TRIM22 SNPs

目的 探讨乙型肝炎病毒(HBV)对肝癌细胞株Huh7细胞TRIM22表达的影响.方法 将Huh7细胞在不同浓度干扰素α(IFN-α)中培养24 h,采用实时定量PCR和Western Blot法检测TRIM22 mRNA和蛋白质表达;将HBV表达载体和空载体分别转染Huh7细胞,然后加入1 000 U/mL IFN-α中培养24 h,检测TRIM22 mRNNA和蛋白质表达.结果 Huh7细胞TRIM22 mRNA基础表达值低,可被IFN-α以剂量依赖的方式诱导表达上调,其中1 000 U/mL IFN-α组TRIM22 mRNA相对表达量为(213.29±43.51),明显高于其他组,差异有统计学意义(P<0.05);转染HBV载体组TRIM22 mRNA相对表达量为(43.59±9.81),明显低于空载体组,差异有统计学意义(P<0.05);转染HBV载体组TRIM22蛋白质表达降低.结论 IFN-α可诱导Huh7细胞TRIM22表达,而HBV可抑制TRIM22表达.     

分泌性内皮抑制素真核质粒治疗小鼠肝癌的初步研究

目的 构建并表达分泌性内皮抑制素真核表达质粒,以此对肝癌进行基因治疗。方法 人工合成Ig κ信号肽序列,和内皮抑制素编码序列一起克隆入pcDNA3.1质粒。重组质粒转染上清液作用于ECV304内皮细胞,MTT法检测内皮细胞的增殖。局部注射重组质粒治疗接种在小鼠腿部肌肉内的H_(22)肝癌瘤株,疗程结束后解剖称取瘤重。结果 构建的内皮抑制素真核表达质粒转染上清液可以抑制内皮细胞的增殖抑制率为29.2％。经质粒裸DNA注射治疗后,治疗组瘤重[(1.34±0.96)g]比空载体组[(2.70±0.82)g]和生理盐水组[(3.73±1.41)g]明显减小(P<0.05)。结论 分泌性内皮抑制素真核表达质粒用于H_(22)肝癌的基因治疗有一定效果。     

骨胳肌葡萄糖载体研究进展

骨胳肌是全身利用葡萄糖的最主要组织,葡萄糖跨细胞膜转运是骨胳肌葡萄糖代谢的限速步骤.葡萄糖的跨膜转运是由位于细胞膜上的葡萄糖载体(glu-cose transporter,GLUT)介导并以易化扩散方式实现的.目前已发现5种葡萄糖载体,骨胳肌中存在2种,即GLUT_1和GLUT_(4).GLUT_1主要存在于红细胞、血脑屏障、胎盘等中,也存在于骨胳肌、心肌及脂肪组织中;GLUT_4则只存在于对胰岛素敏感的组织,如骨胳肌、心肌及脂肪组织中.这两种GLUT在骨胳肌细胞中的含量、分布及对葡萄糖的跨膜转运功能的改变,     

Smad7抑制TGF-β1对肝星状细胞α1(Ⅲ)前胶原基因表达的促进作用

目的:探讨在加入TGF-β1的条件下,Smad7过度表达对肝星形细胞-T6(HSC-T6)α1(Ⅲ)前胶原mRNA水平的作用.方法:体外培养HSC-T6细胞,分为正常对照组、TGF-β1对照组、空载质粒组、Smad7质粒组、TGF-β1+空载质粒组和TGF-β1+Smad7质粒组.通过Fugene6介导Smad7质粒转染,继续培养48 h.采用Real time-PCR、RT-PCR方法检测Smad7和α1(Ⅲ)前胶原mRNA水平.结果:TGF-β1+Smad7质粒组、Smad7质粒组Smad7 mRNA水平较正常对照组、空载质粒组显著增高(t=59.43、59.41、54.27及54.25,均t>t0.01,均P<0.01).Smad7质粒组α1(Ⅲ)前胶原基因mRNA表达与正常对照组、空载质粒组相比无明显差异(t=1.25与1.27,均t0.05).但TGF-β1+Smad7质粒组α1(Ⅲ)前胶原mRNAff水平较TGF-β1+空载质粒组和TGF-β1对照组明显降低(t=103.87与45.70,均t>t0.01,均P<0.01).结论:Smad7可显著抑制TGF-β1对HSC-T6ct1(Ⅲ)前胶原mRNA表达的促进作用,而对HSC-T6α1(Ⅲ)前胶原mRNA表达无明显影响.     

柯萨奇病毒B3感染培养大鼠心室肌细胞L-型钙通道mRNA表达的变化

目的:观察柯萨奇病毒B3感染对培养大鼠心室肌细胞L-型钙通道mRNA表达的影响。方法:用柯萨奇病毒B3感染原代培养新生大鼠心室肌细胞,合成大鼠心室肌细胞L-型钙通道各亚单位引物,通过逆转录-聚合酶链反应技术,以β-actin为内参照,测量钙通道各亚单位mRNA表达量在病毒感染前后的变化。结果:病毒感染组心室肌细胞L-型钙通道α_1(4.00±0.07 vs 2.21±0.41,p<0.01),β(2.06±0.06 vs 1.22±0.30,p<0.05)亚单位mRNA表达量较正常对照组显著增加,而α2/δ(4.12±0.19 vs 4.13±0.27,P>0.05)亚单位mRNA表达量变化无统计学意义。结论:柯萨奇病毒B3感染心室肌细胞钙通道mRNA表达量增加,可能是病毒介导心室肌细胞L-型钙电流变化的分子基础。     

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.     

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。     

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。     

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。     

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     

CagA-Hp抗体IgG与胃炎的组织学分析

研究幽门螺杆菌(Hp)感染与胃炎的关系。方法对204例慢性胃炎患者胃粘膜进行观察分析,并测定其中137例Hp阳性患者血清CagA-Hp抗体IgG水平,与组织学对照。结果慢性萎缩性胃炎伴肠上皮化生患者血清CagA抗体IgG明显高于对照组(P＜0.01);其他类型胃炎患者血清CagA抗体IgG水平无明显增高(P＞0.05)。结论CagA-Hp可能是导致慢性萎缩性胃炎伴肠上皮化生的因素之一,对这类患者应密切随访观察。     

慢阻肺急性加重期患者预后的影响因素分析

目的探讨慢性阻塞性肺病急性加重期(AECOPD)患者预后的相关危险因素。方法回顾性调查、收集58例AECOPD患者可能影响其预后的相关因素,并对其分别进行单因素分析。并进行Logistic多元逐步回归进行多因素分析,筛选影响AECOPD患者预后的独立危险因素。结果单因素分析后将结果 P0.1的因素纳入多因素Logistic回归,分析发现是否合并呼吸衰竭、气促程度、白细胞计数、APACHEⅡ、应用抗氧化剂、慢阻肺治疗依从性为影响AECOPD患者预后不佳的独立因素(P0.05)。结论根据AECOPD患者预后的独立危险因素,及早判断,选择合适的后续治疗方案,对提高其生存率及生存质量具有重要意义。