淋病奈瑟菌mtrC膜蛋白基因的克隆和表达

目的 构建淋病奈瑟菌膜蛋白mtrC表达质粒,探讨淋病奈瑟菌耐药的检测和耐药机制.方法 从淋病奈瑟菌标准株中扩增淋病奈瑟菌膜蛋白mtrC基因片段.酶切后插入pET-28a(+),构建重组表达质粒pET-mtrC.通过质粒双酶切和DNA测序证实该重组质粒构建正确.重组质粒转化入大肠杆菌DE3中.经IPTG诱导表达蛋白.结果 经酶切鉴定和测序分析,质粒构建正确.核苷酸序列与GeneBank(U14993)公布的序列相比较,同源性达到99.5%.通过IPTG诱导,SDS—PAGE可检测到约48500大小的融合蛋白,与预测分子量相同.结论 淋病奈瑟菌膜蛋白mtrC原核表达质粒构建和表达成功,为研究其mtrC外排系统的耐药机制打下基础.     

家族性良性天疱疮一家系ATP2C1基因突变的检测

家族性良性天疱疮(familial benign pemphigus),又称Hailey-Hailey病,是一种常染色体显性遗传性皮肤病。目前认为该病为位于染色体3q21-q24上的ATP2C1基因突变所致。我们应用分子生物学方法对1例家族性良性天疱疮的家系进行了ATP2C1基因突变的检测。     

MDM2蛋白和MDM2 mRNA在尖锐湿疣中的表达

目的 研究尖锐湿疣中MDM2蛋白及MDM2 mRNA的表达情况,探讨MDM2在尖锐湿疣增殖及癌变中的作用。方法 分别采用免疫组化染色及原位杂交方法检测外阴尖锐湿疣和正常人皮肤组织中MDM2蛋白及mRNA的表达情况,同时用PCR法检测人乳头瘤病毒(HPV)型别。结果 32例外阴尖锐湿疣中MDM2蛋白阳性表达18例(56.25%),MDM2 mRNA阳性表达22例(68.75%)。MDM2蛋白与mRNA共同阳性表达14例。PCR检测示尖锐湿疣皮损中HPV6/11型28例(87.5%),HPV16/18型4例(12.5%)。在18例MDM2蛋白阳性表达的标本中,HPV6/11型15例,HPV16/18型3例。在22例MDM2 mRNA阳性表达的标本中,HPV6/11型18例,HPV16/18型4例。而正常人皮肤中MDM2蛋白及MDM2 mRNA均未见表达。结论 MDM2的异常表达可能与尖锐湿疣过度增殖及癌变有关。     

硬斑病样基底细胞癌

报告硬斑病样基底细胞癌1例。患者女,47岁。左鼻背外侧斑疹10余年,溃疡及出血3个月。皮肤科检查可见左鼻背外侧象牙白色至淡黄色斑片,中央可见红斑及淡黄色痂。皮损组织病理：真皮内有嗜碱性肿瘤细胞团块,周边细胞栅栏状排列,并可见收缩间隙。诊断：硬斑病样基底细胞癌。     

成人Blaschko皮炎

报告1例成人Blaschko皮炎。患者女,38岁。双侧胸腹部及腰背部带状融合的丘疹及丘疱疹,伴轻度瘙痒4年余。皮肤科检查：双侧胸腹部及腰背部分别见带状密集融合的暗红色丘疹,上覆少许鳞屑。背部皮损行组织病理检查：表皮轻度角化过度伴灶性角化不全,颗粒层及棘层增生、肥厚,皮突下降、增宽,棘细胞间水肿,真皮乳头及真皮浅层血管扩张,血管周围可见以淋巴细胞为主的炎性细胞浸润。诊断：成人Blaschko皮炎。     

早期先天梅毒1例

报告1例早期先天梅毒。患儿女,70d。发热、背部及四肢起暗红色斑伴脱屑1周。患儿及其父母快速血浆反应素环状卡片试验RPR均阳性(1∶16),梅毒微粒凝集试验TPPA阳性。给予水剂青霉素G5万U/kg·d,连续用药14d全身皮疹完全消退。     

抗原致敏树突状细胞抗小鼠白念珠菌系统感染的实验研究

目的研究不同形态白念珠菌致敏的小鼠骨髓来源树突状细胞(DC)对免疫抑制小鼠白念珠菌系统感染的免疫保护作用及所对应的细胞因子改变。方法孢子相和菌丝相白念珠菌在体外分别致敏小鼠骨髓来源的DC(BM-DC),测定混合培养上清IL-12水平;尾静脉回输免疫抑制小鼠体内后,ELISA法测定各组小鼠脾IFN-γ及IL-4水平,并检测肾携菌量。结果DC孢子致敏组上清IL-12水平(380.2±104.13)pg/mL明显高于DC菌丝致敏组和对照组(P<0.05);而DC菌丝致敏组(74.79±23.47)pg/mL与单纯DC培养组上清IL-12水平(19.71±9.21)pg/mL差异无统计学意义(P>0.05)。孢子致敏DC、菌丝致敏DC分别过继免疫小鼠后,前者脾脏IFN-γ水平(269.43±17.34)pg/g明显高于其他组(P<0.05),IL-4水平(6.23±0.37)pg/g则明显低于其他对照组(P<0.05);荷菌一周后孢子致敏DC回输组小鼠肾携菌量(3.58±2.32)×102CFUs与健康小鼠荷菌组比较无统计学意义(P>0.05);其他各组间肾携菌量比较则有统计学意义(P<0.05)。结论尾静脉回输白念珠菌孢子体外致敏的小鼠骨髓来源DC可有效诱导免疫抑制小鼠抗白念珠菌保护性免疫。     

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.     

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.