Exploratory activity: genetic analysis of its modification by scopolamine and amphetamine

Short-term exploratory activity was found to be significantly higher in C57BL/6By than in BALB/cBy inbred mice. Scopolamine reversed the activity levels in these strains. Basal exploratory activity levels and the effects of scopolamine on this behavioral measure assessed in these two strains, their reciprocal F₁ hybrids, their recombinant inbred strains and three C57BL/6 congenic lines permitted characterization of a gene exerting a major effect on short-term exploratory activity [Exa, linked to H(w26), chromosome 4 (LG VIII)] and of a gene modulating the effects of scopolamine in this behavior, [Sco, linked to H-2, chromosome 17 (LG IX)]. Amphetamine exerted opposite effects in relation to those exerted by scopolamine on activity and its action was found to be determined by a polygenic system.     

His-tag不影响RSV重组蛋白G1F/M2的免疫原性

目的：观察His-tag是否影响RSV重组蛋白G1F/M2的免疫原性。方法：PCR扩增G1和F/M2基因片段,插入表达载体pET-His和pET-DsbA-His中,转化E.coli BL21（DE3）,IPTG诱导表达,采用Ni^＋螯合亲和层析法纯化得His-G1F/M2和DsbA-His-G1F/M2,将后者用凝血酶消化,再经Ni＋螯合亲和层析法纯化得G1F/M2,将His-G1F/M2和G1F/M2免疫BALB/c小鼠,用ELISA测定抗体滴度,MTT法测定细胞毒性T细胞活性（CTL）。结果：两种蛋白在BALB/c小鼠中诱导的RSV特异性抗体和CTL活性无显著差异。结论：His-tag不影响RSV重组蛋白G1F/M2的免疫原性。     

Fas cDNA克隆、表达及重组蛋白纯化的研究

目的:获得高质量及充足的Fas重组蛋白。方法:采用PCR构建表达重组质粒,亲和层析纯化Fas重组蛋白,ELISA检测其免疫学功能。结果:①PCR扩增获得目的片段约 470 bp。②Sanger双脱氧链终止法测序表明,该 DNA序列仅 174位由A→G突变,属于同义突变,与已知的Fas膜外区氨基酸完全一致。③重组质粒经IPTG诱导有一蛋白得以表达,其目的融合蛋白占细菌总蛋白的17. 4%,分子量约为 34．3 kD。④经 Factor Xa切割 Fas融合蛋白,纯化获得 Fas重组蛋白,Western印迹出现特异性Fas蛋白带,分子量为21．3kD,与预测结果相吻合。⑤ELISA检测 Fas重组蛋白,具有抗体的免疫学活性。结论:成功地表达并纯化了Fas重组蛋白,解决了Fas不易获得的难题,为研究Fas提供了良好的实验材料。     

用噬菌体肽库筛选重组日本血吸虫线粒体相关蛋白的表位

目的：筛选和鉴定重组的日本血吸虫（中国大陆株）线粒体相关蛋白rSj38的表位。方法：用纯化的rSj338/26GST免疫家兔获得抗rSj338/26GST的多克隆抗体IgG，将抗体进一步纯化，获得抗rSj338单特异多克隆抗体IgG。用纯化抗rSj338抗体对噬菌体12肽库进行5轮免疫学筛选，挑取克隆。采用Western blot免疫识别，核苷酸序列测定分析其获得的表位并与rSj338/26GST进行同源性比较。将获得的不同表位的阳性克隆分别免疫小鼠，并采用Western blot及dot-ELISA方法筛选能刺激小鼠产生较高滴度抗rSj338抗体的阳性克隆，并将阳性噬菌体免疫的小鼠血清对纯化的rSj338/26GST,26GST，日本血吸虫成虫及虫卵抗原进行Western blot识别。结果：经5轮免疫学筛选后挑取的32个克隆，用Western blot方法30个克隆能被抗rSj338抗体识别，核苷酸序列分析发现共有11种表位，与rSj338无一级结构的同源性。经动物免疫初步实验筛选。共获得4个免疫原性较强的阳性克隆，其免疫鼠血清均可识别rSj338/26GST，日本血吸虫成虫及虫卵抗原。结论：获得了4种日本血吸虫中国大陆株线粒体相关蛋白rSj338的表位，均为模拟表位，这将为日本血吸虫病的疫苗研究开辟新的途径。     

Acute changes of coagulation and fibrinolysis parameters after experimental thromboembolic stroke and thrombolytic therapy

Thrombolysis is the only effective pharmaceutical therapy in acute ischemic stroke in humans but has a high risk of intracerebral hemorrhage. We aimed to establish an animal model to study changes of coagulation and fibrinolytic parameters during thromboembolic ischemic stroke and thrombolysis with recombinant tissue plasminogen activator (rt-PA). We used a thromboembolic stroke model in the rat. Animals were treated with rt-PA thrombolysis (n = 10) and compared with untreated (n = 10), sham operated (n = 10) and control animals (n = 20). Coagulation parameters (APTT, PT, TT, fibrinogen, AT III, TAT) and fibrinolytic parameters (t-PA antigen concentration, t-PA activity, PAI-1 concentration, PAI activity, plasminogen, antiplasmin) were measured at two time points (2.5 and 5 h after stroke induction) with a battery of commercially available test kits. We observed an (1) initiation of coagulation and inhibition of fibrinolysis by the operation procedure itself, (2) simultaneous activation of fibrinolysis and its inhibitors after stroke induction and (3) potent initiation of fibrinolysis and consumption of fibrinolysis inhibitors after rt-PA therapy of stroke. We established a model system to monitor coagulation and fibrinolysis during thrombolytic therapy of stroke in the rat. This model may be used to study the influence of these parameters on hemorrhagic stroke transformation and outcome in experimental stroke in future.     

Quantitative trait loci (QTL) for circadian rhythms of locomotor activity in mice

The loomotor activity of male mice (Mus musculus) was monitored by infrared photoelectric beams under three lighting regimens: LD (12 h of light and 12 h of dark), DD (constant dark), and LL (constant broad-spectrum light, 10 lux). Circadian period of locomotor activioty (τ) was compared among 3 inbred strains of mice, C57BL/6J (B6), BALB/c (C), and DBA/2J (D2), and 26 recombinant inbred strains B×D (B6×D2). the τ under both continuous low-intensity light and continuous darkenss varied significantly among strains. Under DD the mean τ was 23.8 h for B6, 23.7 h for D2, and 23.6 h for C. Under LL the mean τ was 25.1 for B6, 23.9 h for D2, and 25.5 h for C. Frequency histograms of the mean τ of 26B×D RI mouse strains (three to seven animals per strain) in either DD or LL and the difference between them, Δτ, had distributions which appeared unimodal, suggesting polygenic inheritances. The narrow-sense heritability determined using 26 strains of B×D RI mice was about 55% for τ and about 38% for both τ in LL and Δτ. An estimated four loci contribute to the variance of τ in constant darkness and five to the variance of τ in constant low-intensity light among the strains studied. Quantitative trait locus (QTL) analysis identified several potential genetic loci associated with τ in constant darkness, τ in constant low-intensity light, and Δτ. The associations of highest probability for each of these traits were theD1Nds4 locus (p<0.001) on mouse chromosome 1, theD5Ncvs52 locus (p<.05) on mouse chromosome 5, and thePmv12 locus (p<.01) at 70 cM on mouse chromosome 5, respectively. A QTL identified for τ was associated (p<.05) with theD2NDS1 marker at 45 cM on chromsome 2 near the Ea 6 marker at 46 cM associated (p<.05) with that reported for the period of wheel running activity in seven C×B RI strains (Schwartz, W. J., and Zimmerman, P.,J. Neurosci. 10:3685 1990).     

Recombinant congenic strains of mice from B10.D2 and DBA/2: Their contribution to behavior genetic research and application to audiogenic seizures

Recombinant congenic strains (RCS) represent a series of related strains, each of which carries a small fraction of the genome of one strain (donor strain) on the genetic background of another strain (background strain). Recombinant inbred strains (RIS) are commonly used to identify major gene segregation and linkage and associations between behavior and quantitative trait loci, whereas recombinant congenic strains (RCS) open other complementary leads. The variability in the reactivity of RCS to a trait is thus the expression of few minor-effect genes originating from the donor strain, because the probability that major genes are present in any one RCS is low. Unlike RIS in which minor-effect genes are often masked by major genes, RCS enable the effects of minor genes to be studied. With our method, for a given trait, an estimate can be made of the gene strength distribution as well as an estimate of the minimal number of genes involved having a certain strength.This study was supported by the Centre National de la Recherche Scientifique (URA 1924 and CSEAL-UPS 44, CNRS), Université René-Descartes, Paris V UFR Biomédicale, and the Fondation pour la Recherche Médicale.     

超声清创联合重组人表皮生长因子治疗肛周脓肿合并感染的效果

目的探讨超声清创联合重组人表皮生长因子对肛周脓肿合并感染患者表皮细胞生长因子(Epidermal growth factor,EGF)、激活素A(Activin A,ACTA)水平及创面血流量的影响。方法选取2018年2月-2020年1月承德医学院附属医院肛周脓肿合并感染患者117例,依据随机数字表法分为对照A组(n=39)、对照B组(n=39)、联合组(n=39)。常规干预基础上对照A组采取超声清创,对照B组采取重组人表皮生长因子,联合组采取超声清创及重组人表皮生长因子。统计三组治疗情况、治疗前后疼痛程度评分、创面血流量及经皮氧分压、EGF、转化生长因子-β(Transforming growth factor-β,TGF-β)、ACTA、C-反应蛋白(C-reactive protein,CRP)水平及临床疗效。结果联合组创面愈合时间、感染控制时间、住院时间分别为(12.39±2.01)d、(5.22±1.56)d、(13.91±2.51)d短于对照A组、对照B组,创面细菌清除率为(89.13±4.64)%高于对照A组、对照B组(P<0.001);治疗后1 d、3 d、7 d、14 d三组VAS评分较治疗前降低,且联合组低于对照A组、对照B组(P<0.05);治疗后1 d联合组创面血流量及经皮氧分压分别为(0.92±0.25)PU、(35.69±3.20)mmHg大于对照A组、对照B组(P<0.05);治疗后1 d联合组血清EGF、TGF-β分别为(0.70±0.11)μg/L、(0.68±0.12)μg/L高于对照A组、对照B组(P<0.05);治疗后1 d联合组血清ACTA、CRP分别为(9.45±3.58)μg/L、(5.71±2.68)mg/L低于对照A组、对照B组(P<0.05);联合组疗效优于A组和B组(P<0.05)。结论联合采取超声清创及重组人表皮生长因子治疗肛周脓肿合并感染,可改善创面愈合及感染控制情况,缓解疼痛程度,增加创面血流量及经皮氧分压,调节EGF、ACTA等水平,提高整体治疗效果。     

重组人干扰素α2b雾化吸入联合远红外止咳贴治疗小儿毛细支气管炎的临床效果

目的观察重组人干扰素α2b雾化吸入联合远红外止咳贴治疗毛细支气管炎的效果。方法选取2018年3月至2020年3月玉林市妇幼保健院收治的80例毛细支气管炎患儿,采用随机数字表法分为观察组与对照组,每组各40例。对照组给予常规综合治疗,观察组在常规综合治疗的基础上给予氧气驱动雾化吸入α2b干扰素联合远红外止咳贴治疗。比较两组治疗后的临床疗效、临床症状消失时间、住院时间以及肺部功能。结果观察组临床总有效率高于对照组,差异有统计学意义(P<0.05);观察组喘憋消失时间、咳嗽消失时间、肺部啰音消失时间、体温恢复正常时间、住院时间均短于对照组,差异有统计学意义(P<0.05);观察组治疗后潮气量、达峰时间比、达峰容积比均高于对照组,差异有统计学意义(P<0.05)。结论雾化吸入干扰素α2b联合远红外止咳贴治疗毛细支气管炎对提高治疗有效率,缩短临床症状消失时间、住院时间,改善肺功能具有重要作用,值得临床推广使用。     

人的重组促黄体生成素对小鼠卵母细胞体外受精的影响

目的探讨人的重组促黄体生成素(r-hLH)对性成熟前的小鼠卵母细胞体外受精(invitrofertilization,IVF)后的分裂、以及IVF胚的体外培养(invitroculture,IVC)过程中发育状况的影响.方法选用性成熟前的清洁级昆明小鼠,分别用r-hFSH(人的重组促卵泡素20IU),r-hFSH+r-hLH(各20IU)或20IUr-hLH对其进行超排处理,通过对从各处理组小鼠卵巢中手工器械分离出来的卵丘-卵母细胞复合体(cumulusenclosedoocyte,CEO)进行体外受精之后,进一步观察这些CEO经IVF后的受精分裂状况,以及IVC后的发育情况.结果r-hFSH+r-hLH处理组小鼠卵母细胞IVF率(72.2%)极显著高于r-hFSH单独处理组(39.9%)或r-hLH单独处理组(37.2%)的相应指标;r-hFSH+r-hLH处理组和r-hFSH单独处理组的IVF胚,发育至8～16细胞阶段的比例(8～16细胞率)趋于一致(25.15%∶27.18%),但是都极显著高于r-hLH处理组的8～16细胞率(17.24%);r-hFSH处理组IVF胚发育至16～32细胞阶段的总比例(16～32细胞率∶33.01%),极显著高于r-hFSH+r-hLH处理组及r-hLH单独处理组的指标(1.75%和0).结论在IVF条件下,r-hLH对小鼠卵母细胞的受精有明显的促进作用,同时却对相应IVF胚的进一步发育产生明显的抑制作用.