细胞裂解法对单细胞PCR扩增效率的影响

目的研究细胞裂解法对单细胞PCR扩增效率的影响,寻求较好的细胞裂解方法.方法应用显微分离技术分离单个淋巴细胞,分别使用冻融变性和蛋白酶K裂解法处理单细胞,应用多重PCR同时扩增X染色体特异重复序列(DXZ1)和Y染色体特异重复序列(DYZ1)对单细胞进行性别诊断,比较其扩增效率.结果使用显微操作法获得80个男性淋巴细胞.采用冻融变性处理单细胞,40个淋巴细胞经多重PCR扩增后有32个产生DXZ1和DYZ1两条电泳带,正确率为80%.采用蛋白酶K裂解法,40个淋巴细胞经多重PCR扩增后均产生DXZ1和DYZ1两条电泳带,正确率为100%.两者的扩增效率有显著的差异性(χ2=6.81,P<0.01).结论 Rechitsky等的蛋白酶K裂解法比较简便,费时短,扩增效率高,适合细胞裂解及PCR扩增模板的制备.     

Expression of inducible nitric oxide synthase induced by lipid-associated membrane proteins of Ureaplasma urealyticum is regulated by nuclear factor κB-mediated mechanism in murine macrophages

U.urealyticumisthesmallestprokaryoticorgan ismcapableofself replication.Thetinymicroor ganismcouldbeisolatedfromurogenital,placen tasandtherespiratorytractsofpreterminfants.Moreover,U.urealyticuminfectionmaybein volvedinnon specificurethritis(NSU),prostati tis,postpartumfever,infertility,pelvicinflamma torydisease,neonatalpneumoniaandevenchronic lungdisease(CLD)[1].ItisknownthatU.urealyticumlackscellwallstructureandcontains abundantmembraneproteins,butitspathogenicity isstillunknownclearly.…     

表面磁性膜医用316L不锈钢支架的生物相容性

目的 对表面磁性膜血管内支架进行生物相容性研究,为该支架的临床应用提供实验依据.方法 通过溶血实验、动态凝血时间实验、急性全身毒性实验、皮内刺激实验、细胞毒性实验、热源实验、过敏实验、体内植入实验综合评价表面磁性膜血管内支架的生物相容性.结果 表面磁性膜血管内支架无溶血反应及凝血功能的改变,无急性全身毒性反应,无热源反应,支架材料中不存在致敏性物质;支架材料动物体内植入在初期有轻度的炎性反应,12周后炎性反应基本消失,未见炎性细胞浸润积聚现象.结论 表面磁性膜血管内支架具有良好的生物相容性,其应用于临床具有可行性和安全性.     

Effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 （HSV-1）

To observe the effect of Gardenia extract ZG on the adsorption quantity of herpes simplex virus type 1 (HSV-1) so as to explore the mechanism of its antiviral activity, fluorescein isothiocyanate (FITC) was used as the fluorescent probe to label viruses and heparin sodium was used as control. Meanwhile , the effect of Gardenia extract ZG on the adsorption quantity on the surface of Hep-2 cells was determined by flow cytometry. It was demonstrated that adsorption of HSV-1 on the surface of Hep-2 cells exhibited the character of saturation and specificity and heparin sodium could prevent attachment of viruses on these cells. These results are in accord with those reported previously. It was also proved that the manner of drug-use prior to adsorption or simultaneous use of drug and adsorption was better than adsorption prior to drug-use, and the inhibition rates of the former and latter manner were 84. 76% and 82.92% respectively. Three manners of drug-use with Gardenia extract ZG were all effective to reduce the adsorption quantity of viruses, especially the manner of simultaneous use of drug and adsorption with an adsorption inhibition rate of 68.46% . From the above observation, it is apparent that the mechanism of anti-viral activity of Gardenia extract ZG may be via several steps involved in the HSV-1 adsorption.     

血浆凝血酶调节蛋白在评估血管病变和并发症中的意义

目的探讨血浆凝血酶调节蛋白(PTM)在评估血管病变和并发症中的意义。方法用ELISA检测PTM。结果大血管病变的心脑血管疾病(冠心病、脑梗死和脑出血),PTM为轻度升高,平均水平均在35μg/L以下。微血管病变性疾病(原发性慢性肾小球疾病、SLE尿蛋白阳性者、糖尿病微血管病变、多脏器功能不全综合征和D IC),PTM中重度升高,平均水平全在40μg/L以上,分别与大血管病变的各组疾病比较有显著性差异(P<0.01)。分析病情进展和并发症PTM水平,原发性肾小球慢性疾病的肾功能衰竭组显著高于无肾功能衰竭组(P<0.01),SLE尿蛋白阳性组高于阴性组(P<0.01),糖尿病微血管病变组高于大血管病变组(P<0.01),D IC的PTM水平更是显著高于其他所有组别(P<0.01)。结论检测PTM可评估大小血管病变和并发症,有实用的临床指导意义,并为研究病理生理与血管病变的关系提供新的思路。     

载脂蛋白A-Ⅰ对三磷酸腺苷结合盒转运体A1的影响

目的：以THP-1巨噬细胞源性泡沫细胞为研究对象，观察载脂蛋白A-I对THP-1巨噬细胞源性泡沫细胞胆固醇流出和三磷酸腺苷结合盒转运体A1(ABCA1)的影响,从而探讨载脂蛋白A-I对动脉粥样硬化(As)发生发展的影响。方法:用液体闪烁计数器检测细胞内胆固醇流出, 高效液相色谱分析细胞内总胆固醇、游离胆固醇和胆固醇酯含量, 运用逆转录-多聚酶链反应和Western 印迹分别检测ABCA1 mRNA 与ABCA1蛋白的表达,用流式细胞术检测细胞平均ABCA1荧光强度。 结果:载脂蛋白A-I引起THP-1巨噬细胞源性泡沫细胞总胆固醇、游离胆固醇与胆固醇酯呈时间依赖性减少, 而ABCA1蛋白质水平、细胞平均ABCA1荧光强度以及细胞内胆固醇流出呈时间依赖性增加,但ABCA1 mRNA没有明显变化。巯基蛋白酶抑制剂(leupeptin 和ALLN)增加ABCA1蛋白质水平,而其它蛋白酶抑制剂(pepstatin A、aprotinin及phosphoramiddon)不增加ABCA1蛋白质水平,蛋白体抑制剂(lactacytin)和溶酶体抑制剂(NH4Cl)也不影响ABCA1蛋白质水平。 结论:巯基蛋白酶可降解THP-1巨噬细胞源性泡沫细胞ABCA1蛋白质,而载脂蛋白A-I可阻碍巯基蛋白酶降解ABCA1蛋白质,从而提高THP-1巨噬细胞源性泡沫细胞ABCA1 蛋白质水平, 增加细胞内胆固醇流出, 降低细胞内胆固醇聚积。     

Detection and characterization of gastric carcinoma associated with epstein-barr herpes virus

The present investigation was carried out to estimate the incidence of Epstein-Barr virus (EBV)-associated cases among gastric carcinoma (GC) patients in Russia. Carcinoma specimens from 184 patients with GC treated at the Cancer Research Center were investigated by EBV encoded RNA-1 (EBER-1) in situ hybridization. Seventeen (9.24%) cases showed uniform EBER-1 expression restricted to the carcinoma cells. Hybridized signals were not detected in the non-neoplastic gastric epithelium. EBV involvement was significantly more frequent among males, especially in the tumors belonging to less differentiated types (moderately differentiated tubular adenocarcinomas and poorly differentiated solid adenocarcinomas) and located in the upper stomach (cardia and middle part). Most EBV-positive GCs were characterized by great lymphoid compartment involvement. The findings of the distribution of EBV-positive. GCs by sex, site, and histology are similar to those in Japan; however, the detection rate of EBV-positive cases in Russia is higher than that in Japan (6.7%) and lower than that in the USA (16%).     

视频脑电图监测对小儿癫痫的诊断价值探讨

目的探讨视频脑电图(VEEG)对小儿癫痫的诊断价值。方法对102例临床拟诊为癫痫患儿进行4～8h视频脑电图监测,其中包括清醒、睡眠期记录;分析临床发作和异常放电的关系,异常放电出现的时相。结果监测到临床发作并且同步异常放电者23例,仅发作间期癫痫样放电者53例,两者共76例(76/102,74.51%)结合病史确定为癫痫,其中局灶性52例(52/76,68.42%),睡眠中出现55例(55/76,72.37%)。10例(10/102,9.80%)临床发作但无同步异常放电,判定为非癫性发作。结论视频脑电图可提高儿童癫痫诊断的正确率,有助于癫痫的定位,部分剥夺睡眠有助于癫痫样放电的检出率。     

Selection of Immunogenic Peptide Mimics of Male Worm Origin of Schistosoma Japonicum using Phage Display Peptide Library

Phagepeptidedisplaytechniquehasbecomeaverypopu larmethodofstudyinawidevarietyofresearchworkson accountofitsversatility,simplicityandcosteffectiveness. Ithasbeenwidelyusedforidentificationofproteinsand hasprovidedaconvenientmethodologytoobtainligands fors…     

DNA连接酶及相关检测技术研究进展

核酸检测技术已被广泛应用于分子遗传学、微生物学、免疫学和肿瘤学等方面的临床诊断.聚合酶链式反应（polymerase chain reaction,PCR）技术大大促进了病原体、基因突变和基因型的分子诊断方法的发展,它将在临床诊断与分析中起着越来越重要的作用.