SiO2-Pickering乳液佐剂增强沙眼衣原体Pgp3蛋白免疫效果

目的优化二氧化硅(SiO₂)Pickering(SPE)乳液佐剂,研究其对沙眼衣原体(Ct)Pgp3蛋白的免疫增强效应,为预防Ct感染性疾病提供实验依据。 方法优化SiO₂质量浓度、水油比及超声功率,超声乳化制备SPE佐剂。30只小鼠均分为SPE+Pgp3组(Pgp3蛋白和SPE混合免疫)、Pgp3组(Pgp3蛋白单独免疫)和PBS组。ELISA检测各组小鼠血清抗体水平和脾细胞上清细胞因子水平,流式细胞术检测脾细胞γ干扰素(IFN-γ)、白细胞介素-4(IL-4)水平。 结果SPE最优制备条件为超声功率337 W、水油比10∶2、SiO₂质量浓度1 g/L,最优条件下SPE直径为(300.12±44.26) nm,聚合物分散性指数为0.33±0.12。除PBS组外,其他组均检测到了特异性抗体,SPE+Pgp3组总抗体IgG及其亚类IgG1、IgG2a抗体水平高于Pgp3组,且以IgG2a升高为主(P＜0.05)。SPE+Pgp3组、Pgp3组脾细胞IFN-γ和刺激指数高于PBS组,且SPE+Pgp3组高于Pgp3组(P＜0.05)；IFN-γ主要由CD4＋T细胞和CD8＋T细胞分泌。各组IL-4含量均较低。 结论SPE乳液佐剂具有良好的佐剂效应,能有效增强Pgp3蛋白体液免疫和细胞免疫,且可优势诱导Th1型免疫反应。     

Highly discriminative genotyping of Chlamydia trachomatis using omp1 and a set of variable number tandem repeats

This article reports the development of a method for genotyping Chlamydia trachomatis , using PCR and sequencing of omp1 , supplemented with three new variable number tandem repeat (VNTR) loci of C. trachomatis . Typeability, reproducibility and discriminatory power were assessed using four groups of samples: two groups (I and II) of C. trachomatis -positive patients and their positive partner(s), one group (III) of patients with recurrent or persistent C. trachomatis infections, and one group (IV) comprising samples containing a newly discovered mutant strain with a 377-bp deletion in the cryptic plasmid, the new variant C. trachomatis (nvCT). The VNTR loci (designated CT1335, CT1299, and CT1291) were all single nucleotide repeats chosen for maximal mutability and variation. In the study material, nine variants of CT1335, eight variants of CT1299 and five variants of CT1291 were found. The discriminatory power ( D ) of omp1 in the present material was D _omp1  = 0.69. D s for VNTRs CT1335, CT1299 and CT1291 were 0.53, 0.74 and 0.74, respectively. The resolution power of the omp1 -VNTR assay was 0.94. Stability over time of the VNTRs was investigated and found to be adequate for epidemiological studies. Using this genotyping assay, it was confirmed that the nvCT strain was indeed a clone. These results indicate that, with this novel method, strains of C. trachomatis can be individually identified, and epidemiological associations established.     

尿道灌注治疗衣原体和脲原体性尿道炎

目的：探讨乳糖红霉素和地塞米松尿道灌注治疗沙眼衣原体（ＣＴ）和（或）解腺原体（ＵＵ）性尿道炎的疗效。方法：筛选出１５０例不并发淋病、附睾炎及前列腺炎，有尿道炎症状，尿道分泌物荧光聚酶链反应检查ＣＴ和（或）ＵＵ阳性病例，随机分为３组，每机分为３组，每组各５０例。服药组，口服福爱力红霉素；灌注组，用１％乳糖红霉素和０．５％地塞米松作尿道灌注；对照组，尿道灌注生理盐水。１０ｄ为１个疗程。结果：经过１～２     

5种治疗女性泌尿生殖道淋球菌合并衣原体感染方案的成本-效果分析

目的 :探讨不同药物治疗方案对同一疾病所产生的经济效果。方法 :104例女性泌尿生殖道淋球菌合并衣原体感染病例 ,采用随机分组法分为5组 ,分别给予氧氟沙星 (A组 )、盐酸左氧氟沙星 (B组 )、克拉霉素 (C组 )、阿齐霉素 (D组 )、头孢曲松 +红霉素 (E组 ) ,运用药物经济学成本 -效果分析法对5种治疗方案进行分析。结果 :5种方案的成本分别为351、390、474、633、666元。成本 -效果比按治愈率计算分别为5 7、4 5、5 5、7 6、8 3 ;按有效率计算分别为4 9、4 1、5 2、7 1、7 8。结论 :B、C方案为较佳方案     

The 7.5-kb common plasmid is unrelated to the drug susceptibility of Chlamydia trachomatis

Using a new plaque cloning technique, we obtained unique Chlamydia trachomatis strains, which were confirmed to be free of the 7.5-kb common plasmid and glycogen in inclusions. The inclusions in plasmid-free and glycogen-negative strains were characterized by a target-like bull's eye morphology that appeared with the formation of a central translucent area that began to be seen clearly at 30 h postinoculation. The clear zone was composed of gel materials originating from reticulate bodies (RBs) that were abnormally large in size and irregular in shape, although one could not differentiate the aberrant RBs from normal RBs at early stages of development. The in-vitro susceptibility of these strains to various chemotherapeutic antibiotics was tested by comparison with their parent strains possessing the common plasmid. No difference was detected for any of the antibiotics tested, indicating that the 7.5-kb common plasmid is unrelated to the drug susceptibility of C. trachomatis. Received: August 7, 2000 / Accepted: December 11, 2000     

Underrecognition of Cervical Neisseria gonorrhoeae and Chlamydia trachomatis Infections in the Emergency Department

Objectives : 1) To quantify the frequency of underrecognized Neisseria gonorrhoeae and Chlamydia trachomatis cervical infections in women tested in the ED, 2) to describe and compare the characteristics of those treated and not treated during the initial visit, and 3) to quantify the delay interval until treatment was provided. Methods : A 2-year, retrospective consecutive case series was performed from June 1, 1992, to May 31, 1994. There were 148 women with ≥1 discrete occurrence of culture-proven cervical N. gonorrhoeae or C. trachomatis infection studied. All the patients were evaluated in a university-affiliated, tertiary care hospital-based ED with a large rural referral area. The main outcome measures were the proportions of patients with positive cultures both treated and not treated in the ED, the clinical characteristics of each group, and the proportion remaining untreated or experiencing treatment delays of ≥ 2 weeks after attempted phone, mail, and public health follow-up. Results : Of 157 occurrences of positive cultures for N. gonorrhoeae or C. trachomatis, 86 (53%) were treated with a regimen suggested by the CDC prior to ED release. The proportion of women with isolated C. trachomatis infections that were underrecognized and untreated initially was larger than the proportions with isolated N. gonorrhoeae or combined infections (79% vs 27% and 53%, respectively, p < 0.0001). Women with findings suggestive of advanced disease (history of fever or chills, or examination evidence of temperature >38°C, purulent vaginal discharge, or any uterine/salpinx/ovarian tenderness) were more often recognized and treated with appropriate antibiotics initially (p = 0.02 to <0.00001 for all). After phone, mail, and public health follow-up, treatment could not be documented for 25% of the occurrences, in all cases due to an inability to locate the patient. An additional 20% of the women did not receive treatment for 14–60 days. Conclusions : In this population, both N. gonorrhoeae and C. trachomatis cervical infections are frequently underrecognized in the ED, with isolated C. trachomatis infections associated with significantly higher proportions of underrecognition. Many affected women remain untreated for extended intervals, creating public and individual health risks. Improved point of contact detection, follow-up, and treatment policies are needed to limit these risks.     

Chlamydia trachomatis Infection Is Related to Poor Semen Quality in Young Prostatitis Patients

Background

The impact of chronic prostatitis resulting from Chlamydia trachomatis infection on male fertility is controversial.

Objective

To investigate the correlation between C. trachomatis infection and semen quality in young male patients affected by chronic prostatitis resulting from C. trachomatis infection and to evaluate the correlation between anti–C. trachomatis immunoglobulin (Ig) A against heat shock protein 60 (HSP60), heat shock protein 70 (HSP70), and semen parameters.

Design, setting, and participants

All patients with clinical and instrumental diagnosis of chronic prostatitis underwent microbiological cultures for common bacteria, DNA extraction, mucosal and serum antibody evaluation for C. trachomatis, and semen parameter analysis. Western blot analysis of mucosal anti–C. trachomatis IgA was performed.

Interventions

Subjects were split into two groups: Group A consisted of patients with chronic prostatitis resulting from common bacteria (uropathogens), and group B consisted of patients with chronic prostatitis resulting from C. trachomatis infection.

Measurements

The relationship between C. trachomatis infection and semen parameters as well as the correlation among IgA levels, IgA characterisation, and semen analysis were determined.

Results and limitations

We enrolled 1161 patients (mean age: 36.5 yr). Of these, 707 patients were placed in group A, and 454 were placed in group B. Significant statistical differences were reported between groups in terms of sperm concentration (p < 0.001), percentage of motile sperm (p < 0.001), and normal morphologic forms (p < 0.001). Strong correlations between mucosal anti–C. trachomatis IgA and sperm concentration (p < 0.001) and normal morphologic forms (p < 0.001) were reported. Correlations among positivity to HSP60, HSP70, and sperm concentration (p < 0.003) and normal morphologic forms (p < 0.001) were also reported.

Conclusions

This study demonstrated the role of chronic prostatitis resulting from C. trachomatis in male fertility decrease, highlighting probable immunomediated damage to germinal cells because of C. trachomatis infections.     

肺炎衣原体假定蛋白Cpn0146的定位及其生物信息学分析

目的：对肺炎衣原体假定蛋白Cpn0146进行定位与生物信息学分析。方法：使用PCR方法从肺炎衣原体菌株AR39基因组中扩增Cpn0146,并将其插入到pGEX-6P2构建表达质粒pGEX-6p2-Cpn0146并转化大肠杆菌XL-Blue,IPTG诱导其表达融合蛋白GST-Cpn0146。纯化后的融合蛋白免疫小鼠制备特异性抗体,并应用间接免疫荧光法对肺炎衣原体的Cpn0146进行定位分析,最后利用软件Expasy和antheprot5.0对Cpn0146进行生物信息学分析。结果：成功构建了表达载体pGEX-6p2-Cpn0146和制备了抗Cpn0146的多克隆抗体。间接免疫荧光实验显示Cpn0146位于肺炎衣原体的包涵体膜上。生物信息学分析显示Cpn0146具有较好的亲水性与抗原性。结论：肺炎衣原体假定蛋白Cpn0146为一包涵体膜蛋白,其抗原性和亲水性较好。     

沙眼衣原体热休克蛋白60的B细胞表位预测及分析

目的：分析沙眼衣原体（Chlamydia trachomatis,CT）热休克蛋白60（heat shock protein 60,HSP60）的二级结构并预测其B细胞优势表位,进一步分析其在沙眼衣原体相关自身免疫性疾病发病中的意义。方法：以CT标准株（E/UW-5/Cx）HSP60的完整氨基酸序列为基础,分别采用SOPMA、GOR、nnPredict和HNN四种方法预测CHSP60的二级结构,并结合其跨膜区域、亲水性、表面可及性、抗原性、极性和柔韧性等参数进行综合分析,预测CHSP60可能的B细胞优势表位,并将B细胞优势表位与人HSP60的氨基酸序列进行比对,分析两者之间的同源性,寻找可能与自身免疫性疾病相关的B细胞表位。结果：四种方法对CHSP60二级结构的预测表明,其二级结构中柔性区域以无规卷曲为主,少见转角。其B细胞优势表位可能位于氨基酸79～85、135～140、433～440和526～532区域,而上述序列与人HSP60氨基酸序列比对后,发现其135～140、433～440和526～532区域与人HSP60氨基酸159～164,457～464、549～556区域有较高的同源性。结论：CHSP60可能的B细胞优势表位中,135～140、433～440和526～532区域可能与CT感染相关自身免疫性疾病的发病有关。     

沙眼衣原体pORF5蛋白真核表达系统的构建与鉴定

目的构建沙眼衣原体(Ct)pORF5基因重组质粒pDSRed-C1/pORF5,建立pORF5稳定转染的表达系统并对其进行鉴定。方法PCR法扩增pORF5基因,定向插入pDSRed-C1真核表达载体构建pDSRed-C1/pORF5重组体,重组体经酶切、PCR及测序鉴定后,用脂质体介导的基因转染法,分别将pDSRed-C1/pORF5和pDSRed-C1导入HeLa-229细胞,通过G418筛选获得转入目的基因的阳性细胞克隆;荧光显微镜和Western blot检测pORF5蛋白在细胞中的表达。结果所克隆的pORF5基因片段经测序完全正确;转染pDSRed-C1/pORF5的HeLa-229细胞大部分有pORF5蛋白的表达,而转染空载体pDSRed-C1或未转染的HeLa-229细胞,均未见pORF5蛋白表达。结论成功构建了稳定表达外源新基因pORF5的细胞系,为进一步研究该蛋白的结构功能、阐明Ct的致病机制、研制基因工程疫苗提供了有利的条件。