Construction of Prokaryotic Expression Plasmid of mtrC Protein of Neisseria gonorrhoeae and Its Expression in E. Coli

Summary： In order to provide a rational research basis for detection of resistance of Neisseria gonorrhoeae to antimierobial hydrophobie agents and study on the resistant mechanism of multiple transferable resistance （mtr） efflux system, plasmid pET-28a（＋） encoding mtrC gene was constructed and the related target protein was expressed in Escherichia colt （E. cold DE3. The fragments of mtrC gene of Neisseria gonorrhoeae from the standard strains were amplified and cloned into prokaryotic expression plasmid pET-28a（＋） with restriction endonuelease to construct recombinant pET-mtrC which was verified by restriction endonuelease and DNA sequencing. The recom- binant was transformed into E. coli DE3 to express the protein mtrC induced by IPTG. The results showed mtrC DNA fragment was proved correct through restriction endonuelease and DNA sequencing. hs sequence was 99.5 % homologus to that published on GeneBank （U14993）. A 48.5 kD fusion protein which was induced by IPTG was detected by SDS-PAGE. It was concluded that the construction of prokaryotic expression plasmid of mtrC protein of Neisseria gonorrhoeae was correct and the fusion protein was successively expressed in E. coli.     

淋球菌毒力岛分类及sac-4基因对淋球菌血清抗性作用的研究

目的 研究淋球菌不同基因型别的毒力岛类型及其sac-4基因与淋球菌血清抗性作用的关系.方法 运用NG-mast和正常人血清(NHS)的抗性比较试验,对46株淋球菌从分子水平上进行基因分型和毒力岛分类,并在此基础上探讨毒力岛sac-4基因与淋球菌NHS抗性之间的关系.结果 46株野生株中,GGI阳性菌株占80.43%,可分为含有traG和altA;含带sac-4t基因的traG和altA;只含有traG一部分保守序列的三种毒力岛类型.血清抗性实验表明9株sac-4阳性的淋球菌野生株均不具有血清抗性,而sac-4阴性的2号和11号菌株却产生了部分血清抗性和完全抗性.结论 不同毒力岛类型有着不同的毒力特点和基因型别,淋球菌血清抗性作用与毒力岛sac-4基因并无明显相关性.     

局部5-氨基酮戊酸光动力学疗法治疗老年疣的疗效评价

目的：探讨局部5-氨基酮戊酸光动力学疗法（ALA-PDT）治疗老年疣病的疗效。方法：93例老年疣患者,分别采用局部ALA-PDT治疗63例（A组）,二氧化碳（CO2）激光治疗30例（B组）。结果：A组患者经3次治疗后,痊愈16例（25.4%）,显效25例（39.7%）,有效13例（20.6%）,无效9例（14.3%）,总有效率65.1%,复发率6.3%,术后伤口疼痛3.2%,无其它明显副作用发生;B组治疗1次,痊愈率100%,复发率16.7%;伤口并发感染7例（23.3%）,愈后局部遗留浅表瘢痕9例（30%）,伤口疼痛100%。B组复发率、伤口感染率及局部遗留瘢痕的发生率均显著高于A组（P〈0.05）。结论：ALA-PDT无明显痛苦、无瘢痕形成,且复发率低,适合于头面部多发性、较大面积的皮损。     

Expression of IRF-4 and IBP in skin lesions of patients with psoriasis vulgaris

The expression of the interferon regulatory factor 4 (IRF-4) and the IRF-4-binding pro-tein (IBP) in psoriatic skin lesions was investigated. The expression of IRF-4 and IBP in skin lesions of 20 patients with psoriasis vulgaris were immunohistochemically dectected. Normal skin from 10 healthy people was used as normal control. The study showed that expression of IRF-4 was increased significantly in keratinocytes and inflammatory cells in the lesions of psoriasis vulgaris than that in the normal control. The detection revealed that IBP expression in keratinocytes, lymphocytes, hair follicles, and sebaceous glands in normal skin was significantly lower than that in the lesions of pso-riasis vulgaris (P<0.05). Both IRF-4 and IBP might be involved in the pathogenesis of psoriasis vul-garis.     

内皮素对黑素瘤A375细胞转化生长因子-β1及磷酸化Smad 3蛋白表达的调节

Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.     

127例老年疣患者皮损中人类乳头瘤病毒DNA的检测

老年疣又称脂溢性角化病或基底细胞乳头瘤, 是老年人最常见的良性表皮增生性肿瘤,多见于躯干和面部.由于病理检查中容易将几种老年疣与寻常疣等人类乳头瘤病毒（human papillomavirus,HPV）感染疾病混淆,我们采用PCR技术对老年疣皮损进行了病理检查和HPV的检测,以探讨老年疣与HPV感染的关系.     

Construction of Prokaryotic Expression Plasmid of mtrC Protein of Neisseria gonorrhoeae and Its Expression in E.Coli

Gonorrheais one of the most common venerealdiseases.The wild use of antibiotic againstNeisse-ria gonorrhoeaehas made the drug resistance a bigproblemin the prevention and treat ment of gonor-rhea.Besides the drug resistance mediated bychange in structure of penicillin-binding protein,mutation of DNAgyrase A,chromosome plasmid,another non-specificity multi-drug resistant mecha-nismofNeisseria gonorrhoeae,multiple transfera-ble resistance efflux system,has been paid closeattention to increasin…     

孕酮在淋球菌引起的中性粒细胞炎症反应中的调节作用

目的 研究孕酮在淋球菌引起的中性粒细胞炎症反应中的作用。方法 提取正常人外周血中性粒细胞,根据是否加入孕酮,将其分为孕酮组、淋球菌组、干预组（孕酮 + 淋球菌）及对照组。荧光定量RT-PCR分别测定在0、3、8、12 h各组中性粒细胞中诱导型一氧化氮合酶（iNOS）、TNF-α、IL-1β mRNA含量,并用Western印迹测定iNOS蛋白水平。结果 淋球菌组和干预组中iNOS、TNF-α、IL-1β mRNA表达水平均升高;淋球菌组在8 h达到高峰,以后逐渐下降;干预组三者水平明显低于淋球菌组（P < 0.05）。而孕酮组、对照组各因子含量无明显变化。Western印迹结果显示,淋球菌组和干预组iNOS蛋白表达水平亦升高,前者明显高于后者（P < 0.05）。结论 孕酮下调中性粒细胞iNOS、TNF-α及IL-1β的表达,抑制淋球菌引起的中性粒细胞炎症反应,这一机制可能在女性无症状感染中起作用。     

万乃洛韦治疗带状疱疹疗效观察

盐酸万乃洛韦(alaciclovir hydrochloride)是阿昔洛韦的L-缬氨酸酯的盐酸盐,口服后吸收迅速,并在体内很快转化为阿昔洛韦,从而发挥其抗病毒作用.我们自1998年9～12月,对该药治疗带状疤疹进行了临床观察,并与静脉滴注阿昔治韦或病毒唑治疗作比较,取得良好疗效,现报道如下.