白背飞虱中的Wolbachia和Cardinium双重感染特性

为了明确自然种群白背飞虱Sogatella furcifera中Wolbachia和Cardinium的感染情况以及Wolbachia与其特有的WO噬菌体之间的关系, 以采自中国7个省区9个地点的白背飞虱为研究材料, 运用PCR检测的方法调查了Wolbachia, Cardinium以及WO噬菌体在各飞虱种群中的感染率和组织分布特点。结果表明： 白背飞虱广泛双重感染Wolbachia和Cardinium, 并且都表现出很高的感染率。白背飞虱各种群Cardinium的感染率几乎均为100%; Wolbachia的感染率也较高, 但雌雄虫感染率差异较大, 雌虫的感染率几乎均为100％, 而雄虫的感染率从22.2%~95.0%不等。另外, 通过不同DNA聚合酶、 不同提取方法的对比, 揭示了DNA粗提样品在基于PCR技术的胞内共生菌检测中的不足之处。对白背飞虱头部、 胸部、 腹部、 足和翅5个不同部位组织的检测结果表明, 不仅在含有生殖组织的腹部有这两类共生菌的感染, 在其他非生殖组织中同样也感染了这两类共生菌; 虽然Wolbachia和Cardinium在寄主的各个组织中均有分布, 但是两者在白背飞虱成虫（尤其是雄虫）阶段的动态变化有明显的差异。进一步对Wolbachia宿主特异性WO噬菌体的检测结果表明, 自然种群雄虫中Wolbachia的感染率与不感染个体中WO噬菌体的比率呈明显的负相关。因此推测, 雄虫中Wolbachia感染率相对较低的原因可能是由于Wolbachia基因组中溶原性的WO噬菌体受到某种因素的诱导已转化为裂解性噬菌体。研究结果为进一步揭示Wolbachia和Cardinium双重感染条件下对寄主的生殖调控作用及其机制、 垂直传播规律、 两者之间的相互关系以及进一步的应用研究等方面提供了重要的理论基础。     

2015年至2019年辽宁省某医院血培养病原菌分布及耐药分析

分析2015年至2019年辽宁省人民医院血培养分离菌的科室分布及耐药情况,为临床提供数据参考.利用Whonet5.6软件对2015年至2019年辽宁省人民医院血培养临床数据进行分析.血培养阳性率为12.3％,共分离病原菌1266株,其中革兰阴性菌546株、革兰阳性菌649株、真菌71株;革兰阴性菌主要为大肠埃希菌(Es...     

粪菌移植和益生菌在复发性艰难梭菌感染和炎症性肠病中的应用进展

聚集在人体肠道的菌群对维持机体正常的生理功能具有重要作用,肠道菌群的失调会导致复发性艰难梭菌感染及炎症性肠病等疾病。粪菌移植是将健康供者肠道内的功能菌群转移到患病个体的肠道内,重建患者的肠道微生态环境,从而达到治疗的目的。研究发现,粪菌移植在治疗复发性艰难梭菌感染方面表现出较高的治愈率,同时对炎症性肠病有积极的疗效。益生菌是一类能发挥健康作用的活性微生物,当机体摄入足够数量的益生菌,益生菌能在宿主肠道内定植,可以恢复并维持宿主肠道菌群的平衡。益生菌可作为预防复发性艰难梭菌感染的辅助治疗,同时在缓解炎症性肠病方面也有较好的效果。     

泌尿系感染常见病原菌的分布及耐药性分析

【摘 要】 目的 探讨解放军第44医院住院患者泌尿系感染病原菌的分布及耐药性,为临床合理应用抗菌药物提供参考。方法 通过法国生物梅里埃VITEK 2 compact及ATB-Expression细菌鉴定分析仪对2012年该院收治的698例住院患者送检中段尿标本进行细菌鉴定,并用K-B法对分离病原菌进行体外药敏试验。结果 698例患者中有512例培养阳性,其中女性患者阳性率为81.5%,男性患者阳性率为19.0%,二者比较差异有统计学意义(P＜0.05)。512例阳性患者中段尿标本中共分离出病原菌536株,其中革兰阴性杆菌占68.8%,革兰阳性球菌占22.8%,真菌占8.4%。分离病原菌对常用抗菌药物呈现多重耐药性,革兰阴性杆菌对氨苄西林、哌拉西林、头孢噻吩、复方新诺明的耐药率高于60.0%,对亚胺培南、阿米卡星敏感。革兰阳性球菌对克林霉素、红霉素、青霉素的耐药率高于70.0%,对万古霉素、替考拉宁高度敏感。结论 泌尿系感染病原菌对常用抗菌药物耐药严重,及时了解泌尿系感染病原菌的分布特点及耐药性,对临床合理选用抗菌药物具有重要意义。     

8805例外科术后患者医院感染的调查与分析

目的探讨外科术后患者医院感染的发病情况及细菌分布、耐药性,为临床采取防范措施提供依据。方法以2006年1月1日至2006年12月31日全部手术患者为研究对象,前瞻性地调查患者的医院感染情况,并有样必采,进行微生物鉴定和药敏试验。结果外科术后患者医院感染人次发病率为2.2%,例次发病率为3.1%;鉴定出的226株病原体中,G 菌64株,占28.3%,G-菌126株,占55.8%,真菌36株,占15.9%。按分离菌株数排列,依序为大肠埃希菌、铜绿假单胞菌、热带假丝酵母菌、表皮葡萄球菌、溶血葡萄球菌、鲍曼不动杆菌、肺炎克雷伯菌、鲁氏不动杆菌、粪肠球菌、白假丝酵母菌、阴沟肠杆菌。MRSA和MRCNS检出率分别是66.7%和95.5%,大肠埃希菌等G-杆菌对3代头孢类等多种抗菌药物耐药率≥70%。结论该院外科术后患者医院感染发病率符合一般规律,其病原体以G-菌为主,且对3代头孢类等高档抗菌药物多重耐药。     

A therapeutic antibody and its antigen form different complexes in serum than in phosphate-buffered saline: A study by analytical ultracentrifugation

During the development of protein therapeutics, characterization of the active pharmaceutical ingredient is performed extensively to ensure the stability, safety, and efficacy of the drug. Little is known, however, about the characteristics of protein drugs circulating in the blood. The recent availability of a fluorescence detection system (FDS) in analytical ultracentrifugation (AUC) instruments enables the characterization of fluorescently labeled proteins in biological fluids. AUC provides information about protein size, shape, self-association, and binding while avoiding many limitations associated with size exclusion chromatography. Furthermore, with the specificity and sensitivity of FDS, measurements can be performed at physiological concentrations directly in serum. In the current study, we used omalizumab, an anti-immunoglobulin E (IgE) monoclonal antibody, to demonstrate the potential of using AUC-FDS for the study of a monoclonal antibody and its complexes directly in human serum. Omalizumab properties were essentially unaltered after labeling with the fluorescent dye Alexa Fluor 488. In addition, omalizumab and IgE formed different complexes in serum than in phosphate-buffered saline in terms of both size and affinity.     

乙型肝炎病毒假病毒体外感染树鼩原代肝细胞模型的建立

目的：建立乙型肝炎病毒假病毒（HBVpp）体外感染树鼩原代肝细胞（PTH）模型。方法：通过肝脏原位两步灌注法分离PTH并对其冻存方法进行优化,通过共转染293T细胞生产基于慢病毒包装系统的HBVpp并考察其感染的种属和组织特异性。结果：通过肝脏原位两步灌注法分离了PTH,并优化了PTH冻存液的配方;包装的HBVpp具有与HBV真病毒类似的感染种属和组织特异性。结论：该模型的建立对深化HBV进入肝细胞机制的研究具有重要意义。     

Microtubule destruction induces tau liberation and its subsequent phosphorylation

Neurofibrillary tangle-bearing neurons, a pathological hallmark of Alzheimer’s disease, are mostly devoid of normal microtubule (MT) structure and instead have paired helical filaments that are composed of abnormal hyperphosphorylated tau. However, a causal relationship between tau phosphorylation and MT disruption has not been clarified. To examine whether MT disruption induces tau phosphorylation, stathmin, an MT-disrupting protein, was co-expressed with tau in COS-7 cells. Stathmin expression induced apparent MT catastrophe and tau hyperphosphorylation at Thr-181, Ser-202, Thr-205, and Thr-231 sites. In contrast, c-Jun N-terminal kinase activation, or phosphatase inhibition, led to significant tau phosphorylation without affecting MT structure. These findings suggest that MT disruption induces subsequent tau phosphorylation.     

猪瘟病毒感染猪白细胞凋亡基因表达谱变化的研究

为了了解CSFV感染引起猪细胞凋亡的分子机制,利用基因芯片分析了猪感染CSFV前后基因组转录水平的变化。分离攻毒前和攻毒后猪外周血白细胞(Peripheral blood leucocyte,PBL),提取总RNA,经反转录和体外转录得到cRNA,与Affymetrix猪全基因组cDNA芯片进行杂交分析,筛选差异表达的凋亡基因。结果显示,感染猪的PBLs中共筛选到24个涉及细胞凋亡有关基因发生2倍以上变化,并对差异表达的12个宿主细胞凋亡相关基因,用荧光定量RT-PCR进行了验证。功能分析显示,这些凋亡基因的表达变化与CSFV诱导宿主细胞凋亡的机制有关。本文首次用猪全基因组芯片研究了CSFV感染诱导宿主基因的表达变化,建立了CSFV感染猪细胞凋亡相关基因的表达谱,初步阐释了CSFV感染诱导宿主细胞凋亡的分子机制。     

Effect of MOI ratio on the composition and yield of chimeric infectious bursal disease virus-like particles by baculovirus co-infection: deterministic predictions and experimental results

Virus-like particles (VLPs) are empty particles consisting of virus capsid proteins that closely resemble native virus but are devoid of the native viral nucleic acids and therefore have attracted significant attention as noninfectious vaccines. A recombinant baculovirus, vIBD-7, which encodes the structural proteins (VP2, VP3, and VP4) of infectious bursal disease virus (IBDV), produces native IBD VLPs in infected Spodoptera frugiperda insect cells. Another baculovirus, vEDLH-22, encodes VP2 that is fused with a histidine affinity-tag (VP2H) at the C-terminus. By co-infection with these two baculoviruses, hybrid VLPs with histidine tags were formed and purified by immobilized metal affinity chromatography (Hu et al., 1999). Also, we demonstrated that varying the MOI ratio of these infecting viruses altered the extent of VP2H incorporated into the particles. A dynamic mathematical model that described baculovirus infection and VLP synthesis (Hu and Bentley, 2000) was adapted here for co-infection and validated by immunofluorescence labeling. It was shown to predict the VLP composition as a dynamic function of MOI. A constraint in the VP2H content incorporated into the particles was predicted and shown by experiments. Also, the MOI ratio of both infecting viruses was shown to be the major factor influencing the composition of the hybrid particles and an important factor in determining the overall yield. ELISA results confirmed that VP2H was exhibited to a varied extent on the outer surface of the particles. This model provides insight on the use of virus co-infection in virus-mediated recombinant protein expression systems and aids in the optimization of chimeric VLP synthesis.