中西医结合治疗对单纯疱疹病毒性角膜炎患者视力恢复及血清微量元素的影响

目的:探讨中西医结合治疗对单纯疱疹病毒性角膜炎(HSK)患者视力恢复及血清微量元素的影响。方法:选取2015年1月至2016年10月内蒙古包钢医院收治的HSK患者86例作为研究对象,按照随机数字表法随机分为对照组和观察组,每组43例。对照组采用更昔洛韦滴眼液及硫酸软骨素滴眼液治疗,观察组在对照组基础上结合清肝明目汤进行治疗,2组均连续治疗4周。比较2组患者临床疗效;比较治疗前后2组患者视力恢复状况、血清微量元素及泪液免疫因子水平。结果:治疗后,观察组治疗有效率显著高于对照组,差异有统计学意义(P0.05);治疗后观察组患者视觉模拟评分及视力均显著高于治疗前与对照组,差异有统计学意义(P0.05);治疗后观察组铁离子、钙离子及铜离子水平低于治疗前与对照组,差异有统计学意义(P0.05);血清锌离子水平明显高于治疗前与对照组(P0.05),差异有统计学意义(P0.05);治疗后观察组患者泪液IgA、IgG及C3水平明显高于治疗前与对照组,差异有统计学意义(P0.05)。结论:中西医结合治疗HSK有效促进患者视力的恢复,调节血清微量元素的同时增强患者泪液免疫因子水平,疗效显著优于单用西药治疗。     

高糖饮食对小鼠真菌性角膜炎的影响

目的　探究高糖饮食对小鼠真菌性角膜炎的影响。方法　选取健康无眼疾的雄性C57BL/6J小鼠78只，随机分为高糖饮食组和模型对照组，每组36只，模型对照组给予正常饮用水，高糖饮食组给予含体积分数10%果糖溶液，每2 d测量两组小鼠体质量及血糖，10 d后建立真菌性角膜炎模型。造模后24 h、36 h、48 h、72 h、96 h、120 h、168 h裂隙灯显微镜下对角膜进行临床评分并拍照。处死小鼠后，取角膜组织进行HE染色和PAS染色；测定角膜内中性粒细胞和巨噬细胞浸润体积。利用酶联免疫吸附实验对小鼠角膜内的白细胞介素-1β含量进行测定。结果　造模后 0~14 d，两组小鼠体质量与血糖差异均无统计学意义（均为P>0.05）。造模后24 h、36 h、48 h、120 h、168 h，高糖饮食组小鼠角膜临床评分均明显高于模型对照组，差异均有统计学意义（均为P<0.05）。高糖饮食组小鼠角膜穿孔率79.5%，高于模型对照组的40.9%，差异有统计学意义（P=0.000）。造模后各时间点，高糖饮食组中性粒细胞浸润体积均高于模型对照组，差异均有统计学意义（均为P=0.000）。造模后72 h、96 h、120 h、168 h，高糖饮食组巨噬细胞浸润体积均高于模型对照组（均为P=0.000）。角膜组织病理学检查结果示，高糖饮食组炎症反应更重，角膜组织破坏更早且更为严重。造模后24 h、48 h高糖饮食组白细胞介素-1β含量均明显高于模型对照组（均为P<0.05）。结论　高糖饮食加重了真菌性角膜炎感染的严重程度，增强了中性粒细胞、巨噬细胞的趋化，促进了IL-1β的分泌。     

Paradox of complex diversity: Challenges in the diagnosis and management of bacterial keratitis

Bacterial keratitis continues to be one of the leading causes of corneal blindness in the developed as well as the developing world, despite swift progress since the dawn of the “anti-biotic era”. Although, we have expeditiously developed our understanding about the different causative organisms and associated pathology leading to keratitis, extensive gaps in knowledge continue to dampen the efforts required for early and accurate diagnosis, and management in these patients, resulting in poor clinical outcomes. The ability of the causative bacteria to subdue the therapeutic challenge stems from their large genome encoding complex regulatory networks, variety of unique virulence factors, and rapid secretion of tissue damaging proteases and toxins.In this review article, we provide an overview of the established diagnostic techniques and therapeutics for keratitis caused by various bacteria. We extensively report the recent in-roads through novel tools for accurately diagnosing mono- and poly-bacterial corneal infections. Furthermore, we outline the recent progress by our groups and others in understanding the sub-cellular genomic changes that lead to antibiotic resistance in these organisms. Finally, we discuss in detail, the novel therapies and drug delivery systems in development for the efficacious management of bacterial keratitis.     

铜绿假单胞菌噬菌体PaP3生物学特性的研究

目的测定铜绿假单胞菌噬菌体PaP3的最佳感染复数、一步生长曲线和吸附K值及交叉吸附K值等基本生物学特性。方法按照感染复数（MOI）分别为0．0001、0．001、0．01、0．1、1和10加入噬菌体纯培养液和宿主菌，充分裂解细菌后，测定噬菌体滴度；以MOI=10的比例加入噬菌体及宿主菌，进行一步生长实验；纯化PaP3颗粒，免疫家兔，获得抗血清，通过中和反应实验测定PaP3和其抗血清之间的吸附反应常数K值。同时，利用抗血清交叉中和试验确定本室分离的三株噬菌体之间的血清学关系。结果当MOI=0．001时PaP3感染其宿主菌产生的子代噬菌体滴度最高；根据一步生长实验结果绘制一步生长曲线；通过血清交叉中和反应得出不同的吸附常数。结论PaP3最佳感染复数为0．001，感染宿主菌的潜伏期是20min，爆发期是60min，平均爆发量约为31，其抗血清反应的吸附常数K值为262。     

噬菌体基因组PaP1的鸟枪法测序及其生物信息学分析

目的对分离的铜绿假单胞菌噬菌体PaP1进行全基因组测序，并进行初步的生物信息学分析，为噬菌体的改造及其治疗奠定基础。方法采用鸟枪法随机测序和重叠群组装的策略对PaP1进行基因组测序，并通过EditSeq、开放读码框架（ORF）finder、GeneMark^TM、BPROM、FindTerm、Palindromes、Equicktandem及FAStRNA等软件对所获得的基因组序列的一般特征、蛋白质同源性序列及tRNA基因等进行分析和预测。结果PaP1基因组约为90000bp，其中最大重叠群为28249bp。在已获得序列中预测出120个ORFs、41个推定基因及9个簇集在5’末端的tRNA基因。在41个推定基因中，预测出编码DNA末端酶大亚单位、DNA解旋酶B亚单位、肽聚糖结合蛋白及3个尾丝蛋白等6个基因的功能。结论鸟枪法只能测出噬菌体基因组PaP1的部分序列，重复序列的存在是该基因组测序困难的原因。     

Development and evaluation of a new PCR assay for detection of Pseudomonas aeruginosa D genotype

This report describes a new PCR-based assay for the detection of Pseudomonas aeruginosa genotype D in occupational saturation diving systems in the North Sea. This genotype has persisted in these systems for 11 years (1993-2003) and represents 18% of isolates from infections analysed during this period. The new PCR assay was based on sequences obtained after randomly amplified polymorphic DNA (RAPD)-PCR analysis of a group of isolates related to diving that had been identified previously by pulsed-field gel electrophoresis (PFGE). The primer set for the D genotype targets a gene that codes for a hypothetical class 4 protein in the P. aeruginosa PAO1 genome. A primer set able to detect P. aeruginosa at the species level was also designed, based on the 23S-5S rDNA spacer region. The two assays produced 382-bp and 192-bp amplicons, respectively. The PCR assay was evaluated by analysing 100 P. aeruginosa isolates related to diving, representing 28 PFGE genotypes, and 38 clinical and community P. aeruginosa isolates and strains from other species. The assay identified all of the genotype D isolates tested. Two additional diving-relevant genotypes (TP2 and TP27) were also identified, as well as three isolates of non-diving origin. It was concluded that the new PCR assay is a useful tool for early detection and prevention of infections with the D genotype.     

Association between commensal bacteria and opportunistic pathogens in the dental plaque of elderly individuals

Opportunistic infections in the oral cavity of the elderly may increase the incidence of systemic disease. The objective of this study was to investigate the differences in the oral bacterial flora between dependent elderly (inpatients) and independent elderly (community-dwelling residents). After multiple variables were taken into account, inpatients had significantly lower detection rates than community-dwelling residents for alpha-streptococci (p < 0.001) and Neisseria (p 0.004), and higher detection rates for Pseudomonas aeruginosa (p 0.024), methicillin-resistant Staphylococcus aureus (MRSA) (p 0.011) and Actinomyces spp. (p 0.005). Among inpatients, the requirement for a high degree of care was related negatively to detection of alpha-streptococci, but was related significantly to detection of P. aeruginosa (p 0.018) or MRSA (p 0.004). Tube-fed inpatients had a significantly lower detection rate for alpha-streptococci (p 0.041) and a higher detection rate for P. aeruginosa (p 0.004) than those who did not require tube feeding. Inpatients with a history of antibiotic use had a significantly lower detection rate for alpha-streptococci (p 0.049) and a higher detection rate for MRSA (p 0.007) than those without a history of antibiotic use. The detection rates for P. aeruginosa or MRSA in inpatients without alpha-streptococci were higher than in inpatients with alpha-streptococci after controlling for age and gender (P. aeruginosa, p 0.006; MRSA, p 0.001). Overall, detection of alpha-streptococci had an inverse correlation with the detection of P. aeruginosa and MRSA in the oral cavity and is likely to be an indicator of pathogenic bacterial infection.     

用生物发光法定量测定绿脓杆菌对聚苯乙烯的粘附

本文用生物发光技术建立了定量测定绿脓杆菌(PA)对聚苯乙烯表面粘附的研究方法,同时观察了影响 PA 粘附的某些因素,比较了12株 PA 菌株对聚苯乙烯的粘附率。结果表明:在生物发光(BL)反应体系中,在萤虫素/萤虫素酶浓度给定的条件下,发光强度与 PA 释放的 ATP 量呈剂量依赖关系;在 PA 粘附体系中,其与聚苯乙烯表面的粘附随菌量的增加和粘附时间的延长而增加并受其缓冲液 pH 和菌龄的影响;不同 PA 菌株在同一条件下对聚苯乙烯的粘附率具有很大差别。     

聚合酶链反应检测铜绿假单胞菌感染性角膜溃疡的实验研究及临床应用

目的：探索聚合酶链反应（PCR）技术对铜绿假单胞菌感染性角膜溃疡快速诊断的可行性。优越性及其在临床应用中的价值。方法：建立PCR检测标准铜绿假单胞菌方法，对兔铜绿假单胞菌感染性角膜溃疡模型研究并应用于临床，并与细菌培养做比较。结果：铜绿假单胞菌扩增出一条长504bp的阳性条带，其他常见细菌，人和兔正常角膜组织均为阴性，动物模型PCR敏感性为93.8%。特异性为100.0%；细菌培养敏感性为68.7%。特异性为93.7%。临床标本PCR阳性率为66.7%；细菌培养阳性率为38.1%。结论：PCR技术对快速检测实验室和临床标本中的铜绿假单胞菌具有重要的应用价值。