血浆microRNA实时荧光定量PCR检测方法的建立

目的建立特异、稳定且可靠的血浆microRNA(miRNAs)实时荧光定量PCR技术。方法收集10例健康人血浆标本,采用mirVanaTM PARIS kit试剂盒法提取血浆miRNAs,以miRNA特异的茎环引物引导逆转录,通过SYBR Green实时荧光定量聚合酶链反应(PCR)对外参照cel-miR-39、cel-miR-238及血浆miR-342-3p进行检测。结果健康人血浆miR-342-3p及外参cel-miR-39、cel-miR-238可以实现特异性扩增及定量。溶解曲线显示10例健康人血浆标本中cel-miR-39、cel-miR-238和miR-342-3p扩增产物分别在81.44、81.62、82.71℃时出现单一峰,无引物二聚体峰和其他非特异峰出现。血浆miR-342-3p批内批间重复性实验的标准差在0.13~0.20、变异系数(CV)在0.42%~0.66%,显示该方法重复性较好。以cel-miR-39、cel-miR-238作为外参照,同一血浆标本miR-342-3p的5次检测结果ΔCt的标准差为0.22、CV为1.68%,说明cel-miR0-39、cel-miR-238可作为血浆miRNAs实时荧光定量PCR检测的稳定外参。结论实时荧光定量PCR技术可作为研究血浆miRNAs较好的技术平台。     

人肠道病毒71型及柯萨奇病毒A组16型VP1蛋白的真核表达

目的：克隆人肠道病毒71型（EV71）及柯萨奇病毒A组16型（CA16）的VP1蛋白编码基因,构建相应真核表达质粒在体外进行重组表达,检测其细胞内定位,为EV71及CA16的疫苗研究提供基础。方法：通过PCR扩增分别获取EV71及CA16的VP1编码序列,插入pcFlag载体,分别构建EV71及CA16 VP1与FLAG标签融合的真核表达质粒;瞬时转染人胚肾293T细胞及横纹肌肉瘤RD细胞,Western blot法检测融合蛋白的表达,免疫荧光检测融合蛋白的细胞内定位情况。结果：测序表明两种来源VP1蛋白的重组表达质粒构建正确;分别转染两种不同的细胞后,均可通过Western blot检测到相应蛋白表达;免疫荧光检测显示两种毒株的VP1均表达在细胞质中。结论：外源融合表达EV71或CA16 VP1蛋白的真核表达质粒构建成功;所构建质粒分别转染细胞后,均可在细胞质内检测到VP1融合蛋白的表达。     

外生性十二指肠平滑肌肉瘤的影像诊断(附6例报告)

目的　评价各种影像检查方法对外生性十二指肠平滑肌肉瘤的诊断价值。方法　外生性十二指肠平滑肌肉瘤 6例 ,均行消化道钡餐造影 ,B超检查 ,CT检查和 /或MR检查和血管造影检查。结果　所有病例均经手术病理证实为外生性平滑肌肉瘤。上消化道造影表现为外压性改变 ,2例可见肠道外不规则钡斑 ,B超显示上腹实性肿块 ,CT、MR显示为实性肿块 ,2例可见肿块与十二指肠相连 ,血管造影表现为多血供肿瘤 ,由胃十二指肠动脉和肠系膜上动脉的分支右、中结肠动脉供血。结论　外生性十二指肠平滑肌肉瘤的诊断应在消化道作钡餐的基础上联合应用CT和 /或MR及血管造影检查。     

Effect of a preparation of exogenous histone administered by different routes on intraspecific aggression in male mice

A study is made of the effect of a preparation of exogenous histone from calf thymus administered intranasally and intraperitoneally on intraspecific aggression in male mice. The histone is shown to exert a short-term suppressive effect on aggression with both routes of administration which manifests itself in a reduced number of attacks. However, the effective doses with intranasal administration are considerably lower than in the case of intraperitoneal injections. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 120, N ^o 9, pp. 271–272, September, 1995 Presented by I. P. Ashmarin, Member of the Russian Academy of Medical Sciences     

双氢青蒿素诱导肿瘤细胞铁死亡及其机制研究

目的阐明双氢青蒿素(DHA)诱导肿瘤细胞铁死亡的作用及其机制。方法利用3,3′,5,5′-四甲基联苯胺(TMB)检测DHA与FeSO_4体外芬顿样(Fenton)反应生成氧自由基(·OH)的能力;MTT法检测DHA对人肝癌HepG2细胞的毒性(包括FeSO_4与去铁胺预处理组)。MTT法考察谷胱甘肽(GSH)与铁死亡抑制剂(Fer-1)对DHA细胞毒性的影响;采用DCFH-DA染料考察DHA(包括FeSO_4预处理组)诱导的细胞内活性氧的生成能力;采用C11-BODIPY581/591与DiO分别考察DHA(包括FeSO_4预处理组)对细胞内脂质过氧化物生成能力以及细胞膜结构的影响;利用谷胱甘肽过氧化物酶4(GPX-4)试剂盒测定DHA(包括FeSO_4预处理组)对HepG2细胞内GPX-4活性的影响。结果 Fe~(2+)能够催化DHA发生芬顿样反应并生成·OH;DHA的半数抑制浓度(IC_(50))为(39.96±8.78)μmol/L,FeSO_4与去铁胺分别能够增加或者降低DHA的细胞毒性;DHA处理后细胞内活性氧含量与脂质过氧化物含量升高,细胞形态变大,细胞膜呈散点状分布并呈现解离状态。FeSO_4预处理组与DHA组相比较进一步增加细胞内活性氧含量与脂质过氧化物含量,并且细胞膜形态完全破坏。FeSO_4能够增强DHA对GPX-4活性的抑制作用。结论 DHA通过芬顿样反应升高细胞内活性氧而最终诱导肿瘤细胞铁死亡。另外,外源性铁可以加速DHA发生芬顿样反应进而加速肿瘤细胞铁死亡的发生与发展。     

从湿论治过敏性鼻炎探賾

过敏性鼻炎属于临床常见的过敏性疾病之一,其病因病机复杂难辨.湿邪伤人多从口鼻而入,不仅影响鼻的生理功能,又内伤五脏六腑,易致过敏性鼻炎发作.文章从湿邪与过敏性鼻炎的相关性进行探讨,认为无论外湿或内湿皆可引发过敏性鼻炎.其旨在充实中医湿邪致病理论的同时,亦为临床防治过敏性鼻炎提供一定的研究方向.     

中药退热煎剂足浴法辅助治疗小儿外感发热的疗效观察

目的：观察中药退热煎剂足浴法对小儿外感发热的临床疗效。方法：采用随机、对照法,将外感发热患儿120例随机分为治疗组和对照组各60例,在常规抗炎抗病毒等治疗基础上,分别进行中药退热煎剂足浴法和冰力降温贴的辅助治疗,采用卡方检验和t检验比较2组的总有效率、治疗后0．5、1、1．5小时体温变化及降温幅度,并观察2组用药后24、48、72小时的退热疗效。结果：2组治疗总有效率分别为治疗组71．67％,对照组46．67％,两者之间差异显著（P〈0．01）;用药1．5小时后治疗组退热效果优于对照组,差异显著（P〈0．01）;2组体温下降幅度比较,治疗组显著高于对照组,差异有统计学意义（P〈0．05）．结论：中药退热煎剂足浴法治疗小儿外感发热,退热快,体温不宜复升,经济、方便且副作用小。     

清营解表法与温病传统辨治体系关系研究

从卫气营血和三焦辨证论治角度探讨清营解表法与温病传统辨治体系的关系。认为运用复杂性辨证思维,结合当今环境、体质的时代特征,动态把握外感病之传变规律而提出的"营热体质外感"及"清营解表法"理论,是对温病传统理论、治法和药物运用原则的突破和创新,丰富了当今外感病的辨证论治内容。     

人精子染色体离体测试系统中S9混合物的应用

在未加和加入体外代谢激活体系S9混合物2种情况下,将人精子用最终浓度分别为20、40、60μg/ml环磷酰胺处理3.5h,然后与去透明带地鼠卵进行异种体外受精,继而制备精子染色体进行核型分析。在未加S9混合物组,环磷酰胺3种剂量处理的染色体结构畸变精子率依次为10％、12％、10％;断裂均数依次为0.22、0.14、0.20;在加入S9混合物组,畸变精子率依次为18％、24％、32％;断裂均数依次为0.38、0.78、1.64.2组间畸变精子率和断裂均数差异显著(P<0.05)。加入S9混合物组,环磷酰胺3种剂量处理的畸变精子率和断裂均数都高于空白对照组(10％,0.12％),其差异具有统计学意义且畸变精子率和断裂均数随着环磷酰胺剂量增加而增高。本研究结果表明,人精子染色体离体测试系统可以用来检测原诱变剂对人精子中遗传物质的诱变效应。但原诱变剂必须经S9混合物代谢成激活型。     

Reproductive and hormonal factors and risk of lung cancer in women: a prospective cohort study

Several lines of evidence suggest that endocrine factors may play a role in the development of lung cancer, but the evidence is limited and inconsistent. We investigated the association of reproductive and hormonal factors with risk of lung cancer in the National Breast Screening Study, which included 89,835 Canadian women aged 40-59 years at recruitment between 1980 and 1985. Linkages to national cancer and mortality databases provided data on cancer incidence and deaths from all causes, respectively, with follow-up ending between 1998 and 2000. Cox proportional hazards models were used to estimate hazard ratios (HR) and 95% confidence intervals (95% CI) for the association between hormonal factors and lung cancer. During a mean of 16.4 years of follow-up, we observed 750 incident lung cancer cases. After adjustment for covariates, parous women were not at increased risk of lung cancer (HR = 1.18, 95% CI 0.94-1.47) relative to nulliparous women; however, there was a modest increase in risk with increasing parity, reaching a HR of 1.42, 95% CI 1.06-1.88 in women who had 5 or more live births (p for trend 0.02). Among parous women, age at first live birth was inversely associated with risk. Women who had their first live birth at age 30 or older were at reduced risk relative to women who had their first live birth below age 23 (HR 0.68, 95% CI 0.50-0.93, p for trend 0.004). These associations did not differ by age at enrollment (40-49 vs. 50-59 years old), but were somewhat strengthened when attention was restricted to never smokers. Ever use of exogenous hormones showed little association with lung cancer risk; however, long-term users of hormone replacement therapy were at slightly increased risk. Our results add to the limited existing evidence that certain reproductive and hormonal factors may be associated with lung cancer risk in women.