A method for identification of vaginal epithelial microflora by the analysis of 16S ribosomal RNA gene

Analysis of 16S ribosomal RNA gene was carried out in 2 DNA samples isolated from vaginal epithelial smears from women suffering from bacterial vaginosis. The composition of vaginal epithelial microbiocenosis in bacterial vaginosis was determined and its significant difference from normocenosis was shown. A laboratory protocol for identification of vaginal epithelial microflora was developed on the basis of 16S ribosomal RNA gene analysis. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Supplement 2, pp. 39–42, April, 2007     

Evaluation of serum cytokine and chemokine levels in dermatitis herpetiformis: a systematic review and meta-analysis

Immunologic Research -     

First example of familial posttransfusion purpura in two PlA1-negative sisters

Posttransfusion purpura (PTP) (platelet count 5000/microliter) was diagnosed in a female patient (never transfused, gravida IV, para IV) 1 week after transfusion for hysterectomy in 1978. She did not respond to pooled random-donor platelets but recovered following a single plasma exchange. Her platelets were PlA1 negative, and her plasma contained potent anti-PlA1. In 1986, her sister (never transfused, gravida III, para III) developed PTP (platelet counts 5-15,000/microliter) following surgery-associated transfusion. She did not respond to pooled random-donor platelets. Platelet-associated IgG was markedly elevated (5365) molecules/platelet; normal, less than 660); her plasma contained a potent platelet antibody with anti-PlA1 specificity. Her platelets were subsequently shown to be PlA1 negative. The platelet count did not rise above 30,000 per microliter, despite 3 days of high-dose methylprednisolone sodium succinate and 2 weeks of prednisone (80 mg/day). Later, her platelet count increased and remained normal after steroids were discontinued. The two sisters proved to be HLA-identical, and each possessed one haplotype carrying the DR3 marker, which has been implicated as a risk factor in neonatal alloimmune thrombocytopenia associated with anti-PlA1.     

构建大肠癌细胞相关基因Nrf3融合蛋白真核表达载体及其体外转染的初步功能分析

目的：应用基因芯片生物信息学分析思路筛查大肠癌细胞相关基因Nrf3，构建融合蛋白真核表达载体，检测其对体外转染癌细胞周期与凋亡的影响。方法：实验于2004—01／2006—07在南方医科大学病理解剖教研室及肿瘤研究所与中国农业大学农业生物技术国家重点实验室完成。①实验材料：大肠癌细胞组织及其配对的正常大肠黏膜各3例，由解放军总医院病理科提供：大肠癌LoVo细胞系由南方医科大学肿瘤研究所提供；肝癌SMMC7721细胞系由解放军军事医学科学院放射医学研究所提供；真核表达载体pEGFP-N1（Clontech公司）。②实验方法：分别应用Excel表、Affymetrix Microarray Suite Software5．0分析软件和STATA7．0分析软件，对基因芯片表达谱数据行交集补集分析、秩和检验及T检验，筛选“最重要的大肠癌细胞差异表达基因＆ESTs”，差异表达P〈0．05，差异倍数〉2。应用文献轮廓法进一步分析确定首选研究基因为Nrf3，提取组织样品总RNA，cDNA合成，PCR扩增产物经琼脂糖凝胶电泳分离后回收纯化，插入T载体中，SalⅠ和BamHⅠ双酶切后连接到pEGFP-N1载体．获得重组pEGFP-N1-Nrf3质粒。LoVo细胞在体外加入含体积分数为0．1小牛血清、100u／mL青链霉素的RPMI-1640培养基，置于37℃、体积分数为0．05的CO2饱和湿度培养箱中，待转染质粒pEGFP-N1-Nrf3与阴性对照质粒pEGFP-N1转染36h后，荧光显微镜观察候选基因亚细胞定位，FCM分选术观察其在体外对LoVo细胞周期和凋亡的影响。同法观察重组pEGFP-N1-Nrf3质粒体外转染对SMMC7721细胞周期和凋亡的影响。结果：①首选研究大肠癌细胞相关基因的确认：多种统计学方法分析获得最重要的大肠癌细胞相关基因17个，文献轮廓法进一步确认Nrf3为首选研究基因。②Nrf3基因表达：RT-PCR法检测Nrf3在3例大肠癌细胞组织均有表达，而在与其相配对的正常黏膜中表达较弱或不表达，与芯片检测结果基本一致；Nrf3在LoVo细胞中有表达。③Nrf3亚细胞定位：重组pEGFP-N1-Nrf3质粒转染的LoVo细胞其绿色荧光主要分布于细胞核内，提示Nrf3可能定位于细胞核内。（少Nrf3在体外对癌细胞周期与凋亡的影响：体外培养36h后与未转染LoVo细胞的空白对照比较，重组pEGFP-N1-Nrf3质粒转染的LoVo细胞S＋G2／M期所占比例明显减低，G2／M期下降尤为显著，G—G，期细胞所占比例明显增加。Nrf3转染作用于LoVo细胞后，未观察到明显“亚G1”峰（即凋亡峰）的出现。Nrf3体外转染的SMMC7721细胞周期及凋亡情况与LoVo细胞基本相似。结论：大肠癌细胞相关基因Nrf3在体外能够抑制LoVo细胞、SMMC7721细胞的DNA合成和有丝分裂，促使癌细胞阻滞于G0/G，期，抑制其生长增殖的同时不影响细胞凋亡，可作为大肠癌细胞候选分子标志物。     

The effectiveness of the Safety in Interventional Radiology (SIR) Shield in reducing droplet transmission and its effect on image quality and radiation dose

Objective:To evaluate the efficacy of a barrier shield in reducing droplet transmission and its effect on image quality and radiation dose in an interventional suite.Methods:A human cough droplet visualisation model in a supine position was developed to assess efficacy of barrier shield in reducing environmental contamination. Its effect on image quality (resolution and contrast) was evaluated via image quality test phantom. Changes in the radiation dose to patient post-shield utilisation was measured.Results:Use of the shield prevented escape of visible fluorescent cough droplets from the containment area. No subjective change in line-pair resolution was observed. No significant difference in contrast-to-noise ratio was measured. Radiation dosage to patient was increased; this is predominantly attributed to the increased air gap and not the physical properties of the shield.Conclusion:Use of the barrier shield provided an effective added layer of personal protection in the interventional radiology theatre for aerosol generating procedures.Advances in knowledge:This is the first time a human supine cough droplet visualisation has been developed. While multiple types of barrier shields have been described, this is the first systematic practical evaluation of a barrier shield designed for use in the interventional radiology theatre.     

Genetic Heterogeneity of Mycoplasma hominis Clinical Isolates Detected during Observation of Patients with Recurrent Urogenital Inflammation

A rapid reproducible effective method for molecular typing of Mycoplasma hominis strains based on random amplified polymorphic DNA (RAPD) technique was developed. RAPD detected genetic heterogeneity of genomes of Mycoplasma hominis clinical isolates and showed changes in the genomes of Mycoplasma hominis clinical isolates from patients with chronic infection.     

Oral hygiene and associated factors among frail older assisted living residents

The aim of this study was to determine the associations of older assisted living residents’ tooth brushing habits with health and nutritional status. We assessed the tooth brushing habits, nutritional status, oral health, use of dental services, and morbidity of 1,447 assisted living residents in the Helsinki metropolitan area of Finland. Of the residents, 17% did not clean or had not cleaned their teeth and/or dentures daily. Those not cleaning their teeth and/or dentures daily were more often male, less educated and had a mean length of stay in assisted living longer than those who cleaned their teeth and/or dentures daily. They were more often malnourished and dependent in ADLs. In addition they more often had poorer oral health and used fewer dental services. Poor tooth brushing habits indicate poor oral and subjective health. More attention should focus on the oral hygiene of frail older assisted living residents.     

Differing effects of exogenous and endogenous hydrogen sulphide in carrageenan-induced knee joint synovitis in the rat

Background and purpose:

Recent findings suggest that the noxious gas H₂S is produced endogenously, and that physiological concentrations of H₂S are able to modulate pain and inflammation in rodents. This study was undertaken to evaluate the ability of endogenous and exogenous H₂S to modulate carrageenan-induced synovitis in the rat knee.

Experimental approach:

Synovitis was induced in Wistar rats by intra-articular injection of carrageenan into the knee joint. Sixty minutes prior to carrageenan injection, the rats were pretreated with indomethacin, an inhibitor of H₂S formation (dl-propargylglycine) or an H₂S donor [Lawesson''s reagent (LR)].

Key results:

Injection of carrageenan evoked knee inflammation, pain as characterized by impaired gait, secondary tactile allodynia of the ipsilateral hindpaw, joint swelling, histological changes, inflammatory cell infiltration, increased synovial myeloperoxidase, protein nitrotyrosine residues, inducible NOS (iNOS) activity and NO production. Pretreatment with LR or indomethacin significantly attenuated the pain responses, and all the inflammatory and biochemical changes, except for the increased iNOS activity, NO production and 3-NT. Propargylglycine pretreatment potentiated synovial iNOS activity (and NO production), and enhanced macrophage infiltration, but had no effect on other inflammatory parameters.

Conclusions and implications:

Whereas exogenous H₂S delivered to the knee joint can produce a significant anti-inflammatory and anti-nociceptive effect, locally produced H₂S exerts little immunomodulatory effect. These data further support the development and use of H₂S donors as potential alternatives (or complementary therapies) to the available anti-inflammatory compounds used for treatment of joint inflammation or relief of its symptoms.