Heterogeneity of feline herpesvirus type 1 strains

Summary Heterogeneity of 9 feline herpesvirus type 1 (FHV-1) strains consisting of the prototype C27 strain, one French isolate, six Japanese isolates, and the attenuated vaccine F2 strain was examined by biological, immunological, and molecular biological methods. No significant difference was observed in virus growth and antigenic properties among the strains in Crandell feline kidney cell cultures. Hemagglutination activity was also detected in all extracts of cells infected with each strain. However, in immunoblot analysis, a virus-structural immunogenic protein with an M_r of 36 kDa was lacking in 2 strains, one of which was the vaccine F2 strain, whereas the other immunogenic proteins including three kinds of major glycoproteins were detected in all strains without differences in electrophoretic mobilities. Furthermore, when restriction endonuclease analysis was performed to examine the genomic heterogeneity of strains, the cleavage patterns with the enzymeMluI showed a genomic heterogeneity between wild and vaccine strains. In contrast, only a slight variation in the sizes of some fragments was shown with most of the 7 other enzymes used. These results indicated that the lack of the 36 kDa protein and theMluI cleavage pattern could be used as markers of the vaccine F2 strain. The specific markers are important not only to control the quality of the vaccine but also to evaluate the vaccine immunity in FHV-1 infection in cats.     

Expression of canine interferon-gamma by a recombinant vaccinia virus and its antiviral effect

A recombinant vaccinia virus-expressing canine interferon (IFN)-gamma (vv/cIFN-gamma) was constructed. In rabbit kidney (RK13) and canine A72 cells infected with vv/cIFN-gamma, IFN activity was detected in the culture supernatants of both cell types. Canine IFN-gamma was also detected in both cell extracts by Western blot. The activity of the recombinant canine IFN-gamma in RK13 cells was higher than that in A72 cells. The vv/cIFN-gamma could not grow in A72 cells at a low multiplicity of infection, probably due to the antiviral activity of the canine IFN-gamma produced. Although exogenous IFN-gamma did not inhibit the growth of vaccinia virus, addition of anti-canine IFN-gamma serum recovered the growth of the vv/cIFN-gamma on A72 cells in a dose-dependent manner. These results suggest that the growth of vv/cIFN-gamma was inhibited by IFN-gamma produced in a paracrine and autocrine manner. In addition, the recombinant canine IFN-gamma inhibited the multiplication of canine herpesvirus, pseudorabies virus and canine adenovirus type 1 in Madin-Darby canine kidney cells. The antiviral effect of canine IFN-gamma was more effective than that of canine IFN-beta. From the present studies, we concluded the recombinant virus may be a useful suicide viral vector.     

Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay

To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5'-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82- and 52-kDa proteins of B. equi but not those of Babesia caballi. The glutathione S-transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes.     

Construction of Canine Herpesvirus Vector Expressing Foreign Genes Using a lacZ-TK Gene Cassette as a Double Selectional Marker

An improved method for constructing canine herpesvirus (CHV) recombinants expressing foreign genes by using the lacZ-TK gene cassette as a double selectional marker was developed. A recombinant CHV carrying the lacZ-TK gene at a targeted gene locus was constructed and used as a parental virus for generating new recombinants. The parental virus formed blue plaques and was sensitive to TK-specific drugs, while newly generated recombinants, in which the lacZ-TK gene was replaced with the desired foreign gene, become both resistant to the TK-specific drugs and formed white plaques. Recombinants were isolated by using the combination of drug selection and color selection. This improved method allows construction of recombinant CHV with great ease, because the drug selection can enrich the frequency of recombinant CHV from 0.01–0.1% to 10–80%. This method was employed to construct a recombinant CHV that expressed rabies virus (RV) glycoprotein (G protein). This revised version was published online in July 2006 with corrections to the Cover Date.     

遗尿儿童和青少年情绪和行为问题及相关因素研究

目的 调查河南省儿童青少年遗尿症的情绪和行为问题的现况及其影响因素。 方法 从2019年10月—2020年3月,采用分层随机整群抽样方法抽取河南某县5所中小学校,采用一般资料调查表、长处和困难问卷及儿童睡眠调查表进行调查,对影响遗尿患儿情绪和行为的因素进行单因素、多因素分析。 结果 实际发放问卷4 500份,回收4 120份,有效问卷3 912份,有效回收率为94.95%。调查结果显示有7.82%儿童患有遗尿症。遗尿组的情绪症状、多动和困难总分得分明显高于正常组(P＜0.001),两组品行问题、同伴交往和社会行为得分的比较差异无统计学意义(P＞0.05)。Pearson相关分析结果显示,遗尿儿童和青少年的困难总分与打鼾、夜间呼吸问题、嗜睡、注意力缺陷、睡眠总分均存在正相关关系(P＜0.05)。多元线性回归结果显示睡眠呼吸紊乱、照顾者文化程度低、便秘、学习成绩较差是遗尿患儿情绪和行为问题的影响因素。 结论 遗尿患儿情绪和行为问题的检出率高,影响因素是睡眠呼吸紊乱、照顾者文化程度低、孩子学习成绩差、便秘,应引起关注。     

放射卫生机构在核辐射卫生应急体系作用探讨

分析和探讨放射卫生机构在核辐射卫生应急体系中发挥的作用。通过对比发现,核辐射紧急医学救援基地多由取得放射卫生技术服务（甲级）资质或放射性疾病诊断机构资质的单位承担建设任务。放射卫生机构在平时工作中积累的辐射监测、污染检测、剂量估算、健康效应评价等技术能力可在核辐射卫生应急处置中发挥重要作用,实现能力建设的“平急结合”。建议各级放射卫生机构继续发挥自身所长,通过参加放射卫生监测项目的机会提高自身能力,并积极参与放射卫生技术机构检测能力考核,为核辐射卫生应急工作做好人才和技术储备。     

贵州与中国大陆其它地区空肠弯曲菌MLST特征的比较分析

目的 了解贵州省鸡源空肠弯曲菌的分子流行特征,阐明各地菌株间的亲缘关系和进化特征。方法 在贵州省随机抽取7个大型养鸡场,共采集210份鸡的新鲜粪便样本,分离培养出38株空肠弯曲菌,采用多位点序列分型对分离株的序列型及克隆复合体进行构成和聚类分析,与PubMLST数据库中来自国内的分离株分子特征进行比较。结果 贵州省38株鸡源空肠弯曲菌分为23种ST型和8个克隆复合体,其中9种是目前中国大陆地区已有的ST型,2个等位基因和8种序列型为新发现的。ST型构成比前四位分别是ST - 45、ST - 161、ST - 354、ST - 4 324,与数据库中已有的6株贵州来源菌株相比,不存在共同的ST型。优势克隆复合体（CC - 45、CC - 354、CC - 52）与数据库中鸡源分离株的前三位克隆复合体没有重叠。菌株遗传进化树显示,本研究中属于优势克隆复合体的18株菌与国内部分人源、猪源、野生鸟源、鹅源分离株有较近的遗传距离。结论 贵州省鸡源空肠弯曲菌具有高度遗传多样性,在一定程度上表现出地域流行特征。序列型与国内华东地区的分离株有较高的交叉性,部分分离株与国内人、野生鸟、鹅等来源分离株遗传距离近,应加强对不同地区、不同宿主来源空肠弯曲菌的动态监测。     

基于cAMP/PKA/CREB/BDNF信号通路探讨柴胡加龙骨牡蛎汤抗抑郁的作用机制

目的：该研究基于环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/cAMP反应元件结合蛋白(CREB)/脑源性神经营养因子(BDNF)信号通路探究柴胡加龙骨牡蛎汤对慢性不可预见性温和应激(CUMS)法制备的大鼠抑郁模型的干预作用。方法：随机数字表法将60只SD大鼠分为正常组、模型组、柴胡加龙骨牡蛎汤低、中、高剂量组和氟西汀组。除正常组外,其他各组进行49 d的CUMS制备大鼠抑郁模型。第29天开始给药,柴胡加龙骨牡蛎汤组低、中、高剂量组分别给予柴胡龙骨牡蛎汤颗粒剂2.89、5.78、11.56 g·kg^-1,氟西汀组给予盐酸氟西汀胶囊2.06 mg·kg^-1。旷场实验和强迫游泳实验观察大鼠行为学表现,酶联免疫吸附测定法(ELISA)测定大鼠海马组织5-羟色胺(5-HT)、去甲肾上腺素(NE)、cAMP水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠海马组织PKA、CREB、BDNF mRNA表达;蛋白免疫印迹法(Western blot)检测大鼠海马组织PKA、BDNF蛋白表达水平;免疫组化法(IHC)检测CREB定位表达;...