癌组织中miRNA表达谱的变化与NSCLC放疗敏感性的关系

目的观察非小细胞肺癌（NSCLC）患者癌组织中微型核糖核酸（miRNA）表达谱的变化,并探讨其与NSCLC放疗敏感性的关系。方法将30例NSCLC患者根据总体生存率和远端转移情况分为放疗敏感组和不敏感组,采用miRNA表达谱芯片检测癌组织中的miRNA表达谱。结果与放疗不敏感组相比,放疗敏感组中有5个miRNA（miR-126、miR-let-7a、miR-495、miR-451、miR-128b）表达显著上调（P均〈0.01）,7个miRNA（miRNA-130a、miRNA-106b、miRNA-19b、miRNA-22、miRNA-15b、miRNA-17-5p、miRNA-21）表达显著下调（P均〈0.01）。结论放疗敏感和不敏感NSCLC患者癌组织miRNA表达谱有差异,这种差异可能与NSCLC的放疗敏感性有关。     

miRNA在HBV从感染经由肝硬化到肝癌进程中表达谱的变化

目的:分析微小RNA(microRNA, miRNA)在HBV感染到肝硬化再到肝癌进程中表达谱的变化.方法:利用miRNA芯片技术检测人正常肝脏、乙型肝炎肝硬化、HBV相关性肝癌组织中miRNA表达谱的差异. 实时定量PCR检测上述3种肝脏组织中芯片结果差异性表达的miRNA, 验证芯片结果的可信性.结果:与正常肝脏组织相比, 乙型肝炎肝硬化和HBV相关性肝癌组织中表达上调超过2倍的microRNA有6个, 分别为:hsa-miR-602、hsa-miR-129-5p、has-miR-210、hsa-miR-671-5p、hsa-miR-30b*及hsa-miR-572; 表达下调超过2倍的miRNA有8个:hsa-miR-143、hsamiR-199a-5p、has-miR-195、hsa-miR-27a、hsa-miR-99a、hsa-miR-519e、has-miR-130a及hsa-miR-597. 这些正常肝脏组织相比有差异表达的miRNA, 在乙型肝炎肝硬化和HBV相关性肝癌组织中表达量无明显差别.结论:从HBV感染到肝硬化再到肝癌进程中伴有microRNA表达谱的变化, 且变化主要发生在进程早期.     

人肝转移结直肠癌微小核糖核酸表达谱的初步研究

目的 研究人肝转移结直肠癌、无肝转移结直肠癌中微小核糖核酸(miRNA)的表达谱,筛选与结直肠癌肝转移相关的miRNA.方法 收集25例手术切除结直肠癌标本液氮储存.分别选取3例无肝转移和3例肝转移结直肠癌组织,提取组织总RNA,采用illumina miRNA芯片技术检测两种组织中miRNA的表达,筛选两种不同组织中差异表达miRNA,采用实时定量PCR技术在全部结直肠癌组织中对芯片检测结果进行验证.结果 肝转移结直肠癌组织与无肝转移结直肠癌组织中miRNA表达有明显差异,与无肝转移结直肠癌组织相比,在肝转移结直肠癌组织中差异表达的miRNA有28个,其中4个上调,24个下调.肝转移结直肠癌组织中miR-139-3p表达量为1.75±0.40,较无肝转移上调(0.69±0.58,P<0.05),而miR-19a在肝转移中的表达量为0.39±0.20,较无肝转移下调(1.38±0.98,P<0.05),miRNA芯片结果与定量RT-PCR结果一致.结论 miRNA的异常表达可能与结直肠癌的肝转移有关,特异的miRNA表达谱可以为结直肠癌的肝转移提供新的诊断和治疗靶点.     

MicroRNA在人结肠癌干细胞中的表达谱

目的:探讨微小RNA(microRNA,miRNA)在人结肠癌干细胞中的表达,为进一步研究miRNA调控结肠癌干细胞向结肠癌细胞分化的分子机制奠定基础.方法:应用miRNA表达谱芯片检测人结肠癌干细胞和已分化结肠癌细胞中miRNA的表达谱.利用实时定量PCR技术检测两种细胞中差异表达的miRNA,验证miRNA芯片结果的可靠性.应用软件对筛选出的显著差异表达miRNA的靶基因进行预测.结果:与已分化结肠癌细胞相比,人结肠癌干细胞中表达上调超过1.5倍的miRNA有35个.为:hsa-miR-192,hsa-miR-29b,hsa-miR-215,hsa-miR-194,hsa-miR-33a,hsa-miR-32等:表达下调超过1.5倍的miRNA有11个,为:hsa-miR-93,hsa-miR-1231,hsa-miR-524-3p,hsa-miR-886-3p等.PCR技术验证,与miRNA芯片结果相符合.表达显著上调miRNA的共同靶mRNA有:AFF2、MTF1、RUNDC2C和ZFHX4.表达显著下调miRNA的共同靶mRNA有:ONECUT2、SH3TC2、PTPRT、RNABP10、NR3C1、RGSL1、RNASEL和TANC2.结论:筛选出的差异表达miRNA可能参与结肠癌的发病.为该病诊治提供了新的思路.其共同靶基因可能具有重要的调控结肠癌干细胞生长和分化的作用.     

胃癌患者血清中微小RNA分子的特异性表达及其临床意义

目的 探讨胃癌患者血清中微小RNA(miRNA,miR)的表达差异及其与胃癌发生发展的可能关系.方法 用miRNA芯片分析24例胃癌患者及24例健康人血清样本中miRNA表达谱,并采用实时荧光定量聚合酶链反应(qRT-PCR)对差异表达的miRNA在99例胃癌患者及89例健康人血清中进行大样本验证.最后用TargetScan等生物信息学软件分析所筛选miRNA的靶基因.结果 miRNA表达谱芯片分析发现胃癌患者血清中有29条miRNA的表达水平呈现明显变化.qRT-PCR结果表明在胃癌患者血清中miR-27a、miR-370、miR-491-5p和miR-516-3p的表达水平明显下调.生物信息学分析及文献检索发现miR-27a、miR-370、miR-491-5p和miR-516-3p具有肿瘤抑制因子功能,其表达下调与消化道肿瘤的发生发展密切相关.结论 胃癌患者血清中具有特异性miRNA表达谱,miR-27a、miR-370、miR-491-5p和miR-516-3p的表达下调可能在胃癌发生发展中起着重要作用.     

不稳定型心绞痛患者CD4+T淋巴细胞miRNA的差异表达

目的 通过检测不稳定型心绞痛患者循环血CD4+T淋巴细胞基因中miRNA的表达谱,筛选与正常对照者差异表达的miRNA,寻找对CD4+T淋巴细胞具有调控作用的miRNA,为进一步阐明miRNA在不稳定型心绞痛发病机制中的作用提供基础.方法 利用密度梯度离心法分离出不稳定型心绞痛患者和对照者外周血中的单个核细胞,免疫磁珠法进一步分离出CD4+T淋巴细胞.采用miRNA基因芯片技术检测CD4+T淋巴细胞miRNA的表达谱,通过分析软件筛选出不稳定型心绞痛患者和对照者CD4+T淋巴细胞差异表达的miRNA.实时荧光定量聚合酶链反应对差异表达的miRNA进行验证.结果 miRNA基因芯片筛选结果显示,相对于对照者,不稳定型心绞痛患者外周血CD4+T淋巴细胞中表达显著上调的miRNA有miR-155、miR-21、miR-424和miR-127-3p,显著下调的有miR-30b和miR-181a.实时荧光定量聚合酶链反应验证结果与芯片筛选结果一致.结论 筛选得到的不稳定型心绞痛患者循环血CD4+T淋巴细胞miRNA差异表达谱,可能参与了不稳定型心绞痛的发生发展.     

循环microRNAs与局灶节段性肾小球硬化患者疾病活动的关系

目的:分析循环中与局灶节段性肾小球硬化(FSGS)患者疾病活动相关microRNAs (miRNAs),为后续探讨循环miRNAs在FSGS发病中的作用奠定基础. 方法:选取年龄、性别匹配的活动性FSGS患者及正常对照各9例,提取FSGS患者及对照组血浆总RNA,采用TaqMan低密度芯片(TaqMan Low Density Array)进行miRNA表达谱分析,筛选出在活动性FSGS患者循环中表达上调的miRNAs分子;然后再选取32例活动性FSGS患者血浆,采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)方法对筛选出的miRNAs分子进行验证,同时与非活动性FSGS患者进行比较.采用ROC曲线分析循环miRNA区分活动性FSGS、非活动性FSGS和正常对照的效能. 结果:与正常对照相比,活动性FSGS患者循环miRNA-125b,miRNA-186和miRNA-193a-3p水平显著上调(P＜0.05).ROC曲线分析显示,循环miRNA-125b、miRNA-186和miRNA-193a-3p区分活动性FSGS与正常对照的ROC曲线下面积(AUC)分别为0.882、0.789和0.910.与非活动性FSGS相比,活动性FSGS患者循环中miRNA-186水平也明显上调. 结论:活动性FSGS患者循环miRNA-125b、miRNA-186及miRNA-193a-3p较正常对照显著升高,其中miRNA-186的升高与疾病的活动性关系更密切.     

射频消融术改变心房颤动患者外周血微小RNA表达谱

目的探讨心房颤动(房颤)射频消融术是否通过对调控离子通道蛋白的微小RNA(microRNA,miRNA)的影响,实现心房离子流再平衡和逆重构,并试图发现有价值的调控miRNA。方法选择行房颤射频消融术患者(阵发性、持续性和永久性房颤各10例)30例作为房颤组,健康体检者10例作为正常对照组。正常对照组体检时,房颤组射频消融术前和术后3个月分别取外周血,使用miRNA芯片(miRNA v18.0)进行全基因组miRNA表达谱微阵列分析,2组miRNA表达比值≥1.5倍为显著上调,实时定量PCR验证miRNA表达差异结果,并通过mirbase、miranda、targetscan数据库进行靶基因分析。结果与正常对照组比较,房颤组射频消融术前主要参与离子通道蛋白调控的21个miRNA差异表达均有显著意义(P<0.01);房颤组术后3个月与自身术前比较,上述21个miRNA表达亦有明显差异(P<0.01)。其中miR-1266等5个miRNA术前表达上调≥1.5倍,术后明显下调≥10倍;仅miR-3664-5p术前下调8.88倍,术后进一步下降46.06倍;其余15个miRNA均术前表达下调,术后显著上调。结论房颤射频消融术通过影响调控离子通道蛋白的主要miRNA,实现了心房离子流逆重构。调控多个离子流的miR-1266,miR-377-5p,miR-101-5p和miR-151-3p有望成为房颤治疗新靶点。     

蒙古族与汉族高血压病人内皮细胞miRNA表达谱差异分析

目的探讨蒙古族高血压病人和汉族高血压病人血管内皮细胞微小RNA(miRNA)表达谱之间的差异关系。方法将2017年9月—2018年9月赤峰市医院、锡林郭勒盟医院、海拉尔区人民医院就诊的128例蒙古族高血压病人作为观察组,将136例汉族高血压病人作为对照组。比较蒙古族和汉族高血压病人血管内皮细胞miRNA表达谱。结果①蒙古族高血压病人血液中hsa-miR-132、hsa-miR-192、hsa-miR-212、hsa-miR-361-5p、hsa-miR-17、hsa-miR-30、hsa-miR-72、hsa-miR-144、hsa-miR-150、hsa-miR-638、hsa-miR-1249和hsa-miR-185表达上调,hsa-miR-155、hsa-miR-27b、hsa-miR-149、hsa-miR-296-5p、hsa-miR-133b、hsa-miR-1825、hsa-miR-1280、hsa-miR-150表达明显下调。②miRNA差异表达量与高血压分级有关,即高血压分级越高,上调或下调越大,3级高血压最明显。③miRNA-132、miRNA-192、miRNA-212、miRNA-361-5p、miRNA-17、miRNA-155、miRNA-27b的特异度和敏感度较高,可作为蒙古族高血压病人与汉族高血压病人差异的标记物。结论蒙古族与汉族高血压病人血液中miRNA表达存在差异,miRNA-132、miRNA-192、miRNA-212、miRNA-361-5p、miRNA-17、miRNA-155、miRNA-27b的特异度和敏感度较高,可作为蒙古族高血压病人与汉族高血压病人差异的标记物。     

抗结核药物肝损伤与无肝损伤患者化疗前血循环miRNA分子的差异性表达

目的:研究抗结核药物肝毒性(anti-tuberculosis drug-induced hepatotoxicity,ATDH)与非ATDH患者化疗前血浆差异性表达的miRNA分子,为ATDH易感者筛检提供新的生物学指标.方法:采用miRNA芯片检测ATDH及对照患者化疗前血浆标本.对存在差异表达趋势的25种miRNA分子,采用高通量实时荧光定量PCR进行分析验证.应用互联网上miRNA靶基因预测软件对表达上调的miRNA进行靶基因预测,并采用PANTHER tool查找靶蛋白GO功能分类.结果:与对照组相比,ATDH患者化疗前血浆中共检出7个差异性表达的miRNA分子,其中表达上调的miRNA有4个,包括miR-4284、miR-3620、miR-652-5p和miR-4800-5p;表达下调的miRNA有3个,包括miR-338-3p、miR-424-5p和miR-194-5p.结论:ATDH患者化疗前血浆内存在差异性表达的miRNA分子,其中表达上调的miRNA分子(特别是miR-4284)可能作为ATDH易感者筛检的生物学指标.     

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.     

治疗高血压药物的经济学评价

重视高血压治疗中的经济学评价,对利用我国有限的卫生资源来遏制高血压对人民群众的危害有着重要的现实意义。药物经济学对于药物治疗的成本和治疗的结果给予同样的关注。因为治疗高血压的费用,不仅涉及药物价格,还包括患者的危险水平,降压疗效和对临床终点事件的影响,以及治疗的依从性和安全性。因此药物经济学更强调整体成本和价-效比。低危病人,若非药价低廉,治疗的价-效比不够理想。而在高危的患者,价-效比越小越经济而不是药费越便宜越好。