两种乳杆菌联用在食蟹猴阴道的定植效率及其毒性分析

研究乳杆菌鼠李糖乳杆菌-格氏乳杆菌联用制剂(LACT)的定植效率, 评估其有效剂量在动物体内的急性和长期毒性。

在食蟹猴中进行2轮阴道定植试验, 通过检测阴道分泌物pH值和清洁度, 并分离鉴定分泌物中的乳杆菌, 确定乳杆菌联用制剂的定植率和定植剂量; 通过单次和多次阴道给药, 检测联用制剂对食蟹猴体质量、食量、生殖器官及生化指标等的影响, 检测联用制剂在体内的急性和长期毒性。

联用制剂在食蟹猴中连续阴道给药5 d后能成功定植, 改善阴道的微生态; 甲硝唑预处理能够提高定植效率和定植时间; 联用制剂对食蟹猴不具有急性和长期的毒性作用。

在动物模型中, 2种乳杆菌联用制剂对细菌性阴道病具有良好的治疗潜质。

     

人工饲养恒河猴、食蟹猴的繁殖性能初报

目的探索北京地区人工饲养恒河猴与食蟹猴的繁殖性能,为温带地区猕猴的人工饲养和繁殖方式提供借鉴。方法对军事医学科学院实验动物中心饲养的317只恒河猴繁殖群（30只雄猴,287只雌猴）和78只食蟹猴繁殖群（8只雄猴,70只雌猴）近两年的繁殖性状进行观察和统计分析。结果恒河猴母猴妊娠率、繁殖率和成活率分别为60.73%、54.45%和96.89%。食蟹猴母猴妊娠率、繁殖率和成活率分别为79.86%、56.12%和75.00%。结论食蟹猴和恒河猴可以成功的在温带地区饲养和繁殖,但人工饲养食蟹猴的妊娠率与产仔率较恒河猴高,而仔猴成活率则低于恒河猴。     

恒河猴与食蟹猴微卫星DNA多态性的比较分析

目的分析恒河猴和食蟹猴群体间的遗传多样性,确立一种对恒河猴和食蟹猴种群个体的遗传鉴别方法。方法利用聚合酶链反应（PCR）扩增技术采用15个多态性微卫星DNA位点对50只恒河猴和50只食蟹猴个体进行了DNA多态性的分析,对比两群体间等位基因数目差异。结果筛选的15个具有显著多态性的微卫星DNA位点对恒河猴和食蟹猴种群可以进行DNA多态性分析,其等位基因数目均在7个以上,且两群体间有11个位点的等位基因数存在一定的差异。结论利用这些多态性微卫星DNA位点建立一种有效鉴别恒河猴和食蟹猴种群遗传背景的方法具有一定的可行性。     

食蟹猴肾移植模型的建立

目的建立食蟹猴的肾移植模型。方法通过显微外科手术对食蟹猴进行同种异体移植,将供体肾动脉、肾静脉分别端侧吻合到受体腹主动脉、下腔静脉,供体输尿管端端吻合到受体输尿管。结果154例肾移植食蟹猴无一只在手术过程中因意外死亡;无一只因手术感染死亡;无一只因手术并发症死亡;肾热缺血时间（30±4.5）min。结论应用食蟹猴建立肾移植模型,技术成熟,方法可靠。应用此模型评价新型免疫抑制药物,进行免疫耐受、异种移植研究可为临床试验提供更为准确可靠的依据。     

重组人促卵泡激素剂量对食蟹猴卵泡发育和排卵同期化的影响

目的优化食蟹猴胚胎移植同步受体的处理技术。方法选择5~9岁月经周期正常的成年雌性食蟹猴37只,于出现月经血的第2~3天,每日肌注重组人促卵泡激素(rh FSH),按超排剂量和方式分2个实验组,超排供体组、超排受体组,另有一组自然受体组(对照组)。结果和结论 2个实验组超排后卵巢反应全部良好,超排供体组和超排受体组超排后卵巢的获卵总数(15.18±6.51 VS.5.67±3.79)和平均排卵数(6.77±3.61 VS.1.00±0.00)差异具有统计学意义(P<0.05),但超排受体组和超排供体组的雌二醇(E2)水平浓度差异无统计学意义(P>0.05),变化模式一致,且超排受体组与超排供体组排卵同期化的准确率可达83.33%,而对照组仅为42.85%,两者差异具有统计学意义(P<0.05)。     

食蟹猴的基础血糖值调查

目的 调查圈养食蟹猴基础血糖值情况.方法 采用快速血糖仪对153只6～19岁雄性食蟹猴和87只6～24岁雌性食蟹猴的血糖进行测定.结果 不同性别的食蟹猴血糖值存在显著性差异(P＜0.05),其中雌性食蟹猴血糖平均值为4.09 mmol/L±1.03 mmol/L,雄性食蟹猴血糖平均值为3.32 mmol/L±0.59 mmol/L;不同年龄段的食蟹猴血糖值差异显著(P＜0.05),年龄大的食蟹猴血糖值比年龄小的食蟹猴血糖值整体较高;体重指数与基础血糖值之间无显著相关性.结论 食蟹猴基础血糖值与人类基础血糖值相比,水平较低;性别和年龄是影响食蟹猴血糖值的主要因素.食蟹猴基础血糖值调查为糖尿病动物模型的建立及其相关研究提供了有关血糖值的基础数据参考.     

食蟹猴疟原虫红外期裂殖体的组织化学观察

本文对恒河猴肝内食蟹猴疟原虫红外期裂殖体的组织化学成分进行了系统的观察。结果表明,其体内含有糖原、DNA、RNA、蛋白质结合的α-氨基、酪氨酸、色氨酸及组氨酸、碱性蛋白质、网状纤维及胶原纤维、AKP、ACP及ATP酶等生化物质。对各自的定位、含量或活性作了详细描述,并对其生理意义进行了讨论。     

妊娠晚期食蟹猴与同龄未孕食蟹猴血液生理和生化指标的比较

目的提供妊娠晚期食蟹猴有关的血液学数据。方法采用Sysmex XE-2100血液分析仪、SysmexCA-7000凝血分析仪,及OL YMPUS AU600型全自动生化分析仪,对15只怀孕晚期食蟹猴及10只正常对照非孕食蟹猴进行血常规、凝血三项及17项血生化指标分析。结果妊娠晚期食蟹猴在WBC、NEU%、LYM%、PT、APTT、Fgb、TP、Alb、BUN、CR、CHOL、Glu、Fe、LDK、HDBH上,与同龄未孕食蟹猴相比差异显著（P〈0.05）,其他指标RBC、HGB、HCT、MCV、MCH、MCHC、PLT、ALT、AST、ALP、T.Bili、TG、P相比,于孕晚期有不同程度改变,但无显著性差异（P〉0.05）。结论初步建立了孕晚期食蟹猴的常规生理学数据,为相关的实验研究提供基础资料。     

小鼠抗食蟹猴IgG单克隆抗体的制备

目的制备抗食蟹猴、恒河猴等非人灵长类实验动物免疫球蛋白二级抗体,开展对其传染病血清学快速诊断方法的建立。方法采用饱和硫酸铵盐析、Agarose-Protein G亲和层析技术,从食蟹猴血清中提纯IgG。经SDS-PAGE电泳鉴定,采用常规法免疫C57BL/6小鼠,三次免疫后取脾细胞与Sp2/0-Ag14骨髓瘤细胞通过PEG4000融合制备杂交瘤细胞,利用间接ELISA、Western blot等方法进行筛选、鉴定。结果得到5株阳性杂交瘤,分别命名为2B6、2B7、2D9、3B2、5E4,并且5株杂交瘤分泌的抗体均与恒河猴的IgG或血清发生交叉反应,而与其他物种如东北虎、犬等动物的IgG或血清无交叉反应。结论 5株杂交瘤产生的单克隆抗体（McAb）具有较好免疫活性,且能长期、稳定地分泌抗体。此项研究工作为后续研究食蟹猴、恒河猴传染病血清学诊断方法奠定基础。     

食蟹猴糖化血红蛋白水平的调查研究

目的调查食蟹猴群体的HbA1c水平并建立其背景数据库。方法采集后肢静脉血以高效液相色谱法测定176只(雌性88只;雄性88只)食蟹猴的HbA1c值。结果雄性食蟹猴的HbA1c平均值高于雌性食蟹猴(4.80%±0.56%vs.4.61%±0.55%),两者间存在显著性差异(P0.05)。不同年龄段食蟹猴的HbA1c统计结果发现,青年组雌雄性食蟹猴的HbA1c平均值均高于老年组,两个年龄段间的雌性和雄性之间均存在显著性差异(P0.05),而中年组雌性与雄性和其他年龄段间的差异不显著。结论不同性别和年龄段间食蟹猴HbA1c水平具有显著差异性,HbA1c水平与性别和年龄具有相关性。     

Placental transfer of phenylalanine in Macaca mulatta and Macaca fascicularis

We investigated placental transport mechanisms of phenylalanine in Macaca mulatta and Macaca fascicularis. In the beginning of the third trimester we administered i.v. phenylalanine and p-chlorophenylalanine to pregnant animals. Initial higher phenylalanine concentrations were observed followed by a rapid decrease in both rhesus mothers and fetuses when compared with phenylalanine levels in fascicularis mothers and fetuses. In general, however, placental transfer mechanisms of phenylalanine did not differ significantly between the two species.     

Urinary cyclic AMP excretion is increased in Macaca fascicularis and Macaca mulatta compared to humans

Changes in parathyroid hormone and its second messenger cyclic AMP have been implicated in the pathogenesis of osteoporosis. Nonhuman primate models have been useful in the study of osteoporosis, but the physiology of mineral metabolism in certain species is different than in humans. We investigated parameters of mineral metabolism in 15 normal adult female cynomolgus and 14 normal adult female rhesus monkeys. In both species, urinary cyclic AMP was increased compared with humans, and the nephrogenous cyclic AMP in the cynomolgus monkeys was also elevated. Despite this, there was no evidence for hyperparathyroidism in either species as evaluated by serum or plasma phosphorus and midregion-specific and/or aminoterminal-specific immunoreactive parathyroid hormone. Given the increasing use of nonhuman primates in the study of osteoporosis, understanding basic changes in mineral metabolism is important before pathologic effects of bone loss can be understood.     

人工驯养条件下食蟹猴B病毒抗体水平变化规律研究

目的研究人工驯养条件下食蟹猴B病毒抗体水平变化规律,便于有效控制自繁食蟹猴的BV感染率。方法随机选取409只对不同月龄自繁食蟹猴,采用BVELISA法进行BV抗体监测。结果新生仔猴刚出生时均携带不同程度的BV抗体,但随着月龄的增加,BV抗体水平开始下降,至5月龄时BV抗体阳性率降至最低（12．3％）,之后BV抗体水平逐渐升高。结论人工驯养条件下食蟹猴B病毒抗体水平呈由高到低再升高的趋势,5月龄时断奶可最大限度地获得BV抗体阴性猴。     

Social dominance and serum testosterone concentration in dyads of male Macaca fascicularis

The relationship between social dominance and serum testosterone concentration was evaluated in 24 male Macaca fascicularis in 14 dyads at 2-week intervals over an 8-month period. No associations between testosterone concentration and dominance ranks were found in dyads with "clear dominance" relationships. A significant positive association was found between testosterone concentration and dominance ranks in dyads that exhibited "contested dominance" or dominance reversals. Thus, higher testosterone concentration corresponded to social dominance in subjects dominant as the result of a contest, rather than a consequence of high relative rank.     

Prevalence of antibody reaction with cercopithecine herpesvirus 1 antigen in Macaca cyclopis, Macaca fascicularis, and Papio anubis in Taiwan

BACKGROUND AND METHODS: A total of 284 non-human primate sera were collected between December 2004 and September 2005 and tested by a commercially available dot immunobinding assay for the antibodies to cercopithecine herpesvirus 1, an alphaherpesvirus with high mortality for infected humans. RESULTS: Seropositive rates were 58% among non-human primates from animal shelters and 38% among those from zoos and academic institutes. Positive reactors were found in three species, the Formosan macaque (Macaca cyclopis; 57%), the cynomolgus macaque (Macaca fascicularis; 11%) and the olive baboon (Papio anubis; 68%). CONCLUSIONS: Our results showed that natural infection by cercopithecine herpesvirus 1 in Formosan macaques was highly prevalent, and to a certain extent reflected the situation of the wild populations in Taiwan. The findings raised the issues of zoonotic public health and the occupational health of primate workers. High positive rate in olive baboons was also found, although, it cannot be ruled out that the positivity was due to cross-reactivity between cercopithecine herpesvirus 1 and other herpesviruses.     

Comparative studies with six extenders for sperm cryopreservation in the cynomolgus monkey (Macaca fascicularis) and rhesus monkey (Macaca mulatta)

Ejaculated spermatozoa from cynomolgus monkeys and rhesus monkeys were frozen in straws with six different extenders (TTE, DM, mDM, LG-DM, G-DM, and TCG) containing glycerol. Sperm motility and head membrane and acrosomal integrity were evaluated after freezing and thawing, and the cryoprotective effects were compared among the extenders and the two species studied. The results showed that sperm motility and motility recovery with the six extenders were comparable for the cynomolgus and rhesus monkeys. There was no significant difference in sperm motility and head membrane integrity among the six extenders in either the cynomolgus or rhesus monkeys (P>0.05). However, a slightly but statistically lower percentage of acrosomal integrity was found with TCG in both species compared to the other extenders (P<0.05). These findings demonstrate that TTE, DM, mDM, LG-DM, G-DM, and TCG are equally suitable extenders for the cryopreservation of spermatozoa from cynomolgus and rhesus monkeys.     

Social status and reproductive success of male Macaca fascicularis

Dominant males of nonseasonally breeding species should have more opportunity than seasonal breeders to monopolize access to estrous females and thus enhance their reproductive success. They may also use other strategies for maximizing reproductive success such as producing offspring by relatively high ranking females. To test these hypotheses, the paternity of 35 (80%) of 44 Macaca fascicularis offspring, born over a 28-month period, was established using electrophoretically and serologically defined genetic markers. The social ranks of each of the seven potential fathers and 26 potential mothers were determined by recording outcomes of dyadio agonistic interactions. No evidence could be found to support either hypothesis. Further, in view of previously reported significant, positive correlations between male social rank and sexual activity rates in this study group, sexual activity rates do not appear to predict the number of offspring a male sires in this species.     

A comparison of the behavior of Macaca fascicularis and Macaca mulatta in relation to the menstrual cycle

Observations on oppositely sexed pairs of M. fascicularis and M. mulatta revealed certain differences. M. fascicularis showed single-mount copulations, higher potency, less grooming activity, and milder but more frequent aggressive exchanges than the rhesus monkey. These differences might be due to the lower dominance gradient between the sexes and less sexual dimorphism in M. fascicularis than in M. mulatta. There were changes in the sexual, grooming, and aggressive interaction in relation to the menstrual cycle, indicating the suitability of M. fascicularis for hormone-behavior studies. Comparison of data from 30 and 60 min tests showed that important behavioral changes did not become apparent unless animals were given enough time to interact.     

Autonomic nervous system activity in Macaca fascicularis

This study was performed to determine the contributions of the sympathetic and parasympathetic nervous systems to cardiovascular control. Hexamethonium was administered to block the autonomic ganglia, propranolol to block beta adrenergic receptors of the sympathetic nervous system, and methylatropine to block the parasympathetic nervous system. The results of this study indicate high sympathetic tone and low parasympathetic tone in resting Macaca fascicularis. These findings are different from those in man but are similar to other nonhuman primates.