乙酰肝素酶反义寡核苷酸对人胃癌细胞株SGC7901表达bFGF的抑制作用

目的 探讨乙酰肝素酶反义寡核苷酸(AS-ODN)对人胃癌细胞株SGC7901表达碱性成纤维细胞生长因子(basic fibrob-last growth factot，bFGF)的影响。方法 AS-ODN和乙酰肝素酶mRNA起始密码子区互补，无义寡核苷酸(NS-ODN)为对照组，用阳离子脂质体包裹ODNs并转入SGC7901细胞；转染后48小时提取细胞总RNA，半定量RT-PCR法检测乙酰肝素酶基因的含量，免疫细胞化学法检测bFGF的表达。结果 AS-ODN处理后SGC7901表达乙酰肝素酶mRNA下降；处理前bFGF、的表达率为72．23％，不同浓度(0．1、0．2、0．4、0．8μmoL／L）AS-ODN处理后bFGF的表达率下降程度不同(分别为57．15％、51．11％、42．36％和40．25％)。结论 和乙酰肝素酶mRNA起始密码子区互补的AS-ODN对SGC7901表达bFGF有明显影响，且呈剂量相关性。     

MUC2反义脱氧寡核苷酸对胃癌细胞体外黏附侵袭活性的影响

目的 探讨黏蛋白MUC2反义脱氧寡核苷酸（ASODN）对人胃癌细胞株SGC7901黏附侵袭活性的影响。方法 采用人工合成的MUC2 ASODN经阳离子脂质体包裹后转染入SGC7901细胞中,采用黏附试验、Boyden小室体外侵袭试验观察比较转染前后癌细胞黏附率,穿膜细胞相对百分率及组织蛋白酶D、钙黏蛋白表达的变化。结果 转染SGC7901细胞48h后,癌细胞黏附率在30、60、90、120min各时间段逐渐升高,但低于空白对照组（P均〈0．01）;转染后癌细胞穿膜细胞相对百分率明显下降;转染后SGC7901细胞的E-钙黏蛋白表达明显增高,组织蛋白酶D水平明显降低。结论 人工合成的MUC2 ASODN能有效抑制胃癌细胞株SGC7901黏附侵袭能力。     

人核糖体蛋白S13与胃癌细胞多药耐药性的实验研究

目的 探讨人核糖体蛋白S13(RPS13)在胃癌多药耐药(MDR)机制中的作用。方法 采用RT-PCR法扩增RPS13 cDNA片段编码区序列全长，DNA重组技术构建正反义真核表达载体，经脂质体介导转染胃癌细胞SGC7901及胃癌耐药细胞SGC7901/VCR，斑点杂交检测转染细胞mRNA水平的变化；MTT法测定细胞对化疗药物的敏感性，流式细胞仪检测细胞周期。结果 RT-PCR法成功扩增出RPS13 cDNA片段编码区序列全长，并构建正反义真核表达载体；斑点杂交试验证实：正义转染细胞RPS13 mRNA水平上调，反义转染细胞其mRNA水平下调。RPS13正义核酸转染SGC7901细胞后，细胞对阿霉素、5-氟尿嘧啶和长春新碱的敏感性降低；转染反义核酸后，耐药细胞对丝裂霉素和长春新碱的敏感性增加。细胞周期测定表明高表达RPS13后，G1期、S期和G2期细胞的比例分别为47．0％、33．2％和19．8％；低表达RPS13后，G1期、S期和G2期细胞的比例分别为62．9％、1、0％和36．1％。结论 RPS13参与胃癌耐药细胞SGC7901／VCR的多药耐药。     

稳定转染15-PGDH基因对人胃癌细胞生长和迁移的影响

背景：前期研究表明胃癌组织中NAD＋依赖性15-羟基前列腺素脱氢酶（15-PGDH）表达明显减低,瞬时转染15-PGDH对人胃癌细胞的生长和迁移有一定抑制作用。目的：建立稳定转染15-PGDH基因的人胃癌细胞株SGC7901．观察恢复15-PGDH表达对胃癌细胞生长和迁移的影响,探讨15-PGDH与胃癌发生、发展的关系。方法：以双酶切和质粒测序鉴定真核表达质粒pcDNA3／15-PGDH,采用脂质体法将质粒转染人SGC7901细胞,G418筛选稳定转染株．实时聚合酶链反应（PCR）和蛋白质印迹法鉴定。分别以甲基噻唑基四唑（MTT）实验和细胞划痕实验检测稳定转染15-PGDH基因的SGC7901细胞株的增殖情况和迁移能力。结果：经4周G418筛选以及实时PCR和蛋白质印迹法鉴定,得到4株可稳定、较高水平表达15-PGDH的SGC7901细胞株。稳定转染15-PGDH基因的SGC7901细胞株．细胞生长和迁移能力显著低于空质粒组（P〈0．01）。结论：成功建立了稳定转染15-PGDH基因的SGC7901细胞株．恢复15-PGDH表达可显著抑制人胃癌细胞的生长和迁移。15-PGDH表达减低或缺失与胃癌的发生、发展密切相关。     

Ss-A/Ro核蛋白60 ku亚单位反义核酸逆转胃癌细胞多药耐药

目的:对Ro 60反义核酸在逆转胃癌细胞多药耐药中作用进行研究．方法:克隆Ro 60编码基因,构建Ro 60编码基因的反义真核表达载体,将其转导入SGC7901细胞,应用半定量RT-PCR技术,对基因转染细胞进行鉴定,通过MTT法进行体外药物敏感性分析,借助流式细胞仪检测细胞内蓄积的阿霉素．结果:成功构建了Ro 60反义真核表达载体,应用脂质体介导法将其转导入SGC7901-VCR, Ro 60反义真核表达载体转染SGC7901-VCR细胞后,Ro 60的表达量明显下降,体外药物敏感性实验提示其对长春新碱、丝裂霉素、顺铂、阿霉素的敏感性增加,转染反义表达载体的SGC7901-VCR细胞与未转染和转染空载体的细胞相比,IC50值(mg/L)有显著的下降(7．66±0．45 vs 19．56±0．38,17．48±0．85;0．84±0．03 vs 1．62±0．06．1．80±0．03;0．51±0．03 vs 0．87±0．03．0．88±0．03;0．22±0．01 vs 0．52±0．02,0．43±0．03,均P<0．01),细胞内阿霉素蓄积有显著的增加(51．94±1．26 mg／L vs 36．27±0．98,37．01±0．91 mg／L,P<0．01)．结论:Ro 60反义真核表达载体转染SGC7901后能够抑制胃癌细胞的多药耐药表型．     

可溶性耐药相关钙结合蛋白高表达在胃癌细胞耐药中的作用机制

目的 探讨可溶性耐药相关钙结合蛋白(Sorcin)在胃癌耐药中的作用及机制.方法 采用逆转录(RT)-PCR法扩增Sorcin cDNA片段编码区序列全长.DNA重组技术构建FLAG-Sorcin融合表达载体.经脂质体介导稳定转染胃癌SGC7901细胞.RT-PCR及Western印迹法分别检测转染细胞中Sorcin mRNA水平和蛋白水平的表达变化.高效液相色谱(HPLC)法检测Sorcin转染细胞(SGC7901-F-Sor)及对照细胞(SGC7901)内长春新碱(VCR)的浓度及维拉帕米(VRP)对其的影响.结果 RT-PCR法成功扩增出Sorcin eDNA片段编码区序列全长,并构建FLAC-Sorcin融合表达载体.RT-PCR及Western印迹法证实,Sorein在SGC7901转染细胞中高表达.HPLC检测显示Sorcin高表达的SGC7901细胞胞内VCR浓度较其亲本SGC7901细胞降低76.89%.VRP能增加VCR在细胞内的蓄积,VRP处理后,SGC7901-F-Sor细胞内VCR浓度增加2.41倍.结论 Sorcin高表达可致胃癌SGC7901细胞内VCR蓄积减少,提示Sorcin可能通过改变细胞膜两侧药物转运参与胃癌耐药,VRP能部分逆转此作用.     

LKB1对胃癌细胞增殖、凋亡的影响及相关分子实验

目的:探讨LKB1在胃癌发生中的作用及相关分子机制研究,为胃癌化疗药物的研发提供一定的理论基础.方法:利用L K B1过表达质粒以及L K B1s i R N A转染人胃癌细胞株S G C7901,实时荧光定量PCR及Western blot检测质粒以及s i R N A转染效果,流式细胞术检测L K B1过表达以及干扰后的SGC7901细胞(血清饥饿处理2 h)凋亡数量.活性氧检测试剂盒检测L K B1对S G C7901细胞内R O S产生的影响M T T法检测N A C抑制细胞内R O S产生后LKB1过表达以及干扰后SGC7901细胞数量的改变.Western blot检测LKB1过表达以及干扰后SGC7901细胞中凋亡相关蛋白的表达水平.结果:LKB1过表达质粒转染后的人胃癌细胞株SGC7901细胞凋亡增加,胞内ROS产生增加,LKB1过表达后的SGC7901细胞凋亡则明显增多(3.54%vs 1.29%),转染LKB1 si RNA的人胃癌细胞株SGC7901早期凋亡和晚期凋亡的细胞数量占总数较转染scramble si RNA组明显减少(0.54%vs 1.39%)同时Western blot检测发现LKB1过表达后的SGC7901细胞中剪切型Caspase3表达明显升高上升3.12倍,较对照组差异有统计学意义而si RNA干扰后剪切型Caspase3表达则表现为相反的趋势.结论:LKB1通过促进ROS的产生,上调剪切型Caspase3介导的凋亡通路促进胃癌细胞凋亡,因此LKB1在胃癌发生发展过程中具有重要抑制作用,其可能作为胃癌化疗药物研发的重要目标分子.     

Survvin反义寡核苷酸诱导人胃癌细胞凋亡的作用

目的探讨生存素（Survivin）反义寡核苷酸（ASODN）诱导人胃癌细胞SGC7901凋亡的作用。方法设计合成特异性靶向Survivin的ASODN,将胃癌细胞株分为空白对照组（Sham组）、脂质体转染对照组（Lip组）、正义链转染对照组（Lip-SODN组）和ASODN转染组（Lip-ASODN组）。作用48h后,Westemblot法检测各组Survivin表达情况,流式细胞仪检测各组细胞凋亡率,免疫组化SP法检测细胞中PCNA表达情况。结果脂质体介导Survivin ASODN转染后的胃癌细胞Survivin蛋白表达明显下降;ASODN转染组细胞凋亡率明显高于各对照组（P均〈0．05）,各对照组间无统计学差异（P〉0．05）。ASODN转染后胃癌细胞中PCNA表达水平明显降低。结论 Survivin ASODN转染胃癌细胞能下调Survivin蛋白表达,诱导胃癌细胞凋亡,抑制细胞增殖,具有明显的抗癌作用。     

《临床消化病杂志》2005年第17卷总目录

论著血浆置换治疗慢性重型肝炎临床疗效观察1(3)乙酰肝素酶反义寡核苷酸对人胃癌细胞株SGC7901表达bFGF的抑制作用1(5)经内镜氩离子凝固术治疗隆起糜烂性胃炎的研究1(8)肝硬化门脉高压患者血浆SS、VIP及MTL变化的临床意义1(10)酒石酸锑钾诱导人结肠癌细胞凋亡1(12)Smad4和Galectin3在胃癌中的表达及其临床意义1(14)慢传输型便秘患者结肠动力与胃肠激素的关系1(17)应用组织芯片研究凋亡抑制基因Survivin在胃癌中的表达1(20)纳洛酮对酒精性脂肪肝大鼠作用探讨1(23)不同年龄段胃肌电变化的研究1(26)肝硬化患者植物神经功能变化1(28)直肠…     

TGIF对胃癌细胞株SGC-7901的维甲酸信号通路的影响

目的:探讨TG-相互作用因子(TGIF)对胃癌SGC-7901 细胞株中维甲酸信号通路的影响．方法:在TGIF表达质粒稳定转染SGC-7901细胞后,用 Western blot鉴定高表达TGIF的阳性克隆．在TGIF反义寡核苷酸瞬时转染SGC-7901细胞株后,用RT-PCR检测转染效率．再用1 μmol／L ATRA分别处理稳定转染组或瞬时转染组及其对照组细胞,MTT观察细胞增殖速度的变化,流式细胞术观察细胞凋亡率的变化．结果:稳定转染TGIF表达质粒和瞬时转染TGIF反义寡核苷酸对SGC-7901细胞的增殖和凋亡没有明显影响．但在ATRA作用后,TGIF表达质粒转染组细胞的增殖速度比未转染组和空白质粒转染组细胞快(0．434± 0．035 vs 0．386±0．020,0．360±0．014,P<0．05),而细胞的凋亡率的变化比未转染组和空白质粒转染组细胞小,仅由1．14％增加至1．39％．TGIF反义寡核苷酸转染组细胞的增殖速度比未转染组和突变寡核苷酸转染组细胞慢(0．320±0．044 vs 0．388±0．024,0．409±0．041, P<0．05)．而细胞凋亡率的变化比后两组大,由3．09％上升至10．2％．结论:TGIF能拮抗ATRA抑制SGC-7901细胞增殖和诱导其凋亡的作用,TGIF可能参与了对SGC-7901细胞的维甲酸信号通路的抑制．     

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。     

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。     

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。     

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.     

Angiographic diagnosis of the degree of serosal invasion of carcinoma of the colon

Angiography using Prostaglandin El^® was performed on 38 patients with carcinoma of the colon in order to diagnose the degree of serosal cancer invasion. The findings at angiography were classified into four groups:1) AG-S3, abnormal change (irregularity and/or encasement) up to marginal vessels; 2) AG-S2, abnormality up to vasa recta; 3) AG-S1, abnormality of penetrating branches of vasa recta within the wall of the colon; and 4) AG-S0, no distinct findings of abovementioned vessels. These angiographic findings were compared with both macroscopic and microscopic serosal cancer invasion. Angiographic diagnosis is in accord with the macroscopic findings in 84.2 percent of cases. Angiographic diagnosis is in accord with the microscopic findings in 32.4 percent of cases. Macroscopic findings confirm the angiographic diagnosis precisely but the conflict with microscopic findings should not be overlooked. This may be the result of inflammatory change, adhesion, and fibrosis around carcinoma of the colon.