小鼠抗青霉素和头孢菌素酰化酶抗体制备及交叉反应研究

为探讨用不同方法制备的抗原对抗体效价的影响以及抗体之间的交叉反应 ,采用 3种不同方法制备的抗原分别免疫 BAL B/ c小鼠 ,制备出抗青霉素酰化酶 α亚基、β亚基和抗头孢菌素酰化酶α亚基、β亚基 4种抗体 ,测定了它们的效价 ;同时还比较了用同一种方法制备的抗原免疫小鼠后制作这两类抗体的效价以及上述 4种抗体之间对应抗原的交叉反应。结果表明 ,用割胶法制备的抗原免疫小鼠后所得抗体效价较高 ;同样用割胶法制备的抗原免疫小鼠后所得的两类抗体效价无显著差异 ;在交叉反应测试中 ,抗头孢菌素酰化酶抗体也能识别青霉素酰化酶 ,但抗青霉素酰化酶抗体却几乎不能识别头孢菌素酰化酶 ,即抗头孢菌素酰化酶抗体更为活跃 ,而抗青霉素酰化酶抗体则更为专一。同类抗体对应抗原 2个亚基之间的交叉反应说明其互相结合部位结构的类似 ,同时也说明只有进一步制作单克隆抗体才能筛选出特异性强的抗体     

抗精子抗体对鸡繁殖性能的影响

种鸡受精率和孵化率的高低直接影响种鸡场的经济效益。通常影响种鸡受精和孵化的因素有 :种鸡质量、精液品质、输精方法、种蛋保存条件和时间、孵化条件等。很少有人考虑免疫因素引起的受精率下降和孵化率降低。笔者对某种鸡场受精率下降的原因进行试验和分析 ,证实该场种鸡受精率下降主要由母鸡体内的抗精子抗体所致。抗精子抗体的产生直接导致受精率下降 ,甚至不受精 ,并且影响血蛋率和孵化率 ,给种鸡场造成很大的经济损失。1　材料与方法1 .1　试验动物分组　从某鸡场受精率下降鸡群和受精率正常鸡群中选出部分鸡作为试验鸡 ,并分组单笼…     

影响昆明小鼠和BALB/c小鼠胚胎干细胞分离、克隆、传代的若干因素

对昆明小鼠和BALB／c小鼠2种胚胎的研究表明，2种品系小鼠胚胎均可用作胚胎干细胞（ES）分离建系的材料，两者在ES分离、传代上无显著差异（P〉0．05）；大鼠心肌细胞条件培养液与添加LIF的ES常规培养液相比，显著提高了小鼠ES细胞F1、F2出现率（P〈0．05）；采用低浓度消化液使形成ES克隆的比率分别从14％、16％提高到35％、32％，使ES传到第5代的比率分别从1．7％、0提高到5％、7．1％；而采用连续消化法使形成ES克隆的比率从15％、17％提高到40％、50％，使ES传到第5代的比率从0、1％提高到10％、20％；对ES进行伊红染色、核型分析、AKP染色及体外分化能力检测，证实所分离的ES符合小鼠ES的一系列特征。     

昆明小鼠与BALB/c小鼠MHC-Ⅱ分子恒定链的序列差异分析及多克隆抗体的制备

分离昆明小鼠与BALB/c小鼠脾脏淋巴细胞,经Con-A活化,提取细胞总RNA,RT-PCR获得小鼠Ii分子（P31、P41）基因,DNAStar软件分析序列。将BALB/c小鼠P31/P41分子插入到pGEX-KG多克隆位点构建原核表达载体,IPTG诱导并经GST标签蛋白纯化柱纯化表达蛋白,将GST-P31/P41蛋白分别与弗氏佐剂混合乳化后免疫家兔,采集血清。测序分析显示,两品系小鼠P31、P41均分别为648、840bp,编码215个、279个氨基酸;序列比对结果表明,昆明小鼠、BALB/c小鼠P31、P41分子与GenBank已登录核苷酸序列与/或氨基酸序列均存在多个位点差异,且所克隆的两品系小鼠P31分子之间核苷酸序列同源性为98.8%,氨基酸序列同源性为98.6%;两种小鼠P41分子之间核苷酸序列、氨基酸序列同源性均为99.3%;通过与人、鸡对应Ii分子进行比较,发现与人同源性较高,核苷酸序列同源性在79.0%以上,氨基酸序列同源性在73.0%以上,而与鸡同源性相对较低,核苷酸序列同源性在55.0%以上,氨基酸序列同源性在41%以上。IPTG诱导并纯化后获得GST-P31/P41蛋白,免疫家兔后所采集的血清经ELISA、Western-blotting证实获得了兔抗P31/P41多克隆抗体。结果表示,昆明小鼠与BALB/c小鼠Ii分子间存在差异位点,并成功制备了兔抗Ii分子多克隆抗体,为开展靶向性表位疫苗研究奠定基础。     

臭氧对小鼠精子畸形率和睾丸标志酶活性的影响

为了研究臭氧对小鼠精子畸形率和睾丸乳酸脱氢酶(LDH)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)活性的影响,以及臭氧的生殖损害效应,试验采用臭氧发生器产生臭氧,每天分别处理30 min、60 min、120 min,分别在处理后第30天和第60天处死10只小鼠.结果表明:处理第30天和60天时不同臭氧处理组精子畸形率...     

柠檬酸对小鼠精子特性和睾丸组织形态结构的影响

60只雄性小鼠随机分为4组,每天分别腹腔注射0.2mL的0.9%生理盐水或7.5、15、30g/L CA,于14、21d测定曲细精管精子的生成及附睾内精子特性的相关数据。结果显示,随柠檬酸处理剂量的增加精子的生成数减少。14d时,对照组与15、30g/L CA组间、7.5、30g/L CA组间曲细精管内精子的生成差异显著（P〈0.05）;21d时,对照组与柠檬酸处理组间及7.5、15g/L CA与30g/L CA处理组间曲细精管内精子的生成有显著差异（P〈0.05）。柠檬酸对附睾精子品质的影响表现为：对照组与柠檬酸处理组间及柠檬酸处理组间精子密度和活动率差异均显著（P〈0.05）;对照组和15、30g/L CA处理组及15g/L CA与30g/L CA处理组间精子活力差异显著（P〈0.05）;精子的畸形率随柠檬酸浓度的增加而增加,15g/L CA处理组精子畸形率显著高于对照组（P〈0.05）,30g/LCA处理组显著高于对照组和7.5g/L CA（P〈0.05）处理组。组织形态学观察结果表明柠檬酸能破坏睾丸正常组织形态结构。柠檬酸高剂量组的生精小管内生精细胞排列疏松、紊乱,生精细胞间出现空隙;生精小管中的生精细胞脱落或退化,精原细胞、精母细胞和精子数量明显减少。研究表明,外源柠檬酸雄性小鼠具有显著的生殖遗传毒性。     

灌胃不同奶样对BALB/c小鼠致敏性的影响

本试验研究了驼奶、牛奶和人奶对BALB/c小鼠的致敏性强弱,将60只小鼠随机分为5组,每组12只,分别为β-乳球蛋白（β-lg）组（阳性对照组）、牛奶组、驼奶组、人奶组和空白组（阴性对照组）,每组分别灌胃1 mg/g体重的样品和0.3 μg/g体重的霍乱弧菌毒素（CT）,空白组灌胃PBS和CT,每周1次。灌胃6周后,通过观察过敏症状,检测血清特异性免疫球蛋白E（IgE）和IgG1、血浆组胺水平及血管通透性等指标,比较几种奶源的致敏性强弱。结果显示,驼奶组、人奶组和空白组小鼠体重正常增长,而β-lg组和牛奶组的体重增长有减慢的趋势。与空白组相比,β-lg组和牛奶组的血清特异性IgE和IgG1水平极显著升高（P<0.01）,组胺水平极显著升高（P<0.01）,血管通透性增加,过敏症状明显;而驼奶组小鼠血清IgE和IgG1水平极显著低于牛奶组（P<0.01）,但与人奶组相比无显著差异（P>0.05）,过敏症状轻微,表明驼奶的致敏性明显低于牛奶,且与人奶相似。     

小檗胺对牛病毒性腹泻病毒感染BALB/c小鼠的影响研究

试验旨在研究天然小分子化合物小檗胺(berbamine,BBM)对牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)感染BALB/c小鼠的影响.共分为预防保护和治疗两部分试验.预防保护试验将24只BALB/c小鼠随机分成3组,每组8只,使用0、50、100 mg/kg BBM提前灌胃,...     

模拟急性低压性低氧对小鼠部分血气指标的影响

为了阐明在模拟急性低压性低氧环境下血气指标的变化趋势,选取BALB/c小鼠作为研究对象,检测了模拟急性低压性低氧时其血液中动脉与静脉的血氧饱和度、氧分压、二氧化碳分压等血液指标的变化。结果表明：随着缺氧时间的延长,BALB/c小鼠动、静脉血氧饱和度差值逐渐增大,24 h后趋于平衡;动、静脉氧分压差随着缺氧时间的延长表现为先降低后增加的趋势;动、静脉二氧化碳分压差随着缺氧时间延长表现为先增加后降低的趋势。该种动态变化主要是由于BALB/c小鼠随着缺氧时间延长,代偿性提高氧的供应、提高氧的利用效率,达到适应缺氧环境的目的。     

肉碱对冻融猪精子总抗氧化能力、抗氧化酶基因及凋亡相关基因表达的影响

试验旨在探究肉碱对冻融猪精子质量、抗氧化、抗凋亡及精卵结合能力的影响。试验分鲜精组和冻融组,冻融组的肉碱浓度分别为0、0.025、0.05、0.075 mg/mL,对冻融后的精子质量、总抗氧化能力、抗氧化酶相关基因及凋亡相关基因mRNA表达、精卵结合能力进行检测。结果显示,在冻融组中,经过肉碱处理的精子存活率、质膜完整率和顶体膜完整率均显著高于0 mg/mL处理组（P < 0.05）,其中肉碱浓度为0.05 mg/mL时改善效果最好;经过冻融处理的精子中丙二醛（MDA）浓度与鲜精组相比显著升高（P < 0.05）,其中肉碱浓度为0.05 mg/mL时显著低于其他肉碱处理组（P < 0.05）;与鲜精组相比,各冻融组精子总抗氧化能力均显著降低（P < 0.05）,肉碱浓度为0.05 mg/mL时总抗氧化能力最高。此外,与鲜精组相比,0 mg/mL肉碱处理组中凋亡相关基因Caspase-3与Bax的相对表达量显著升高（P < 0.05）,抗凋亡基因Bcl-2的相对表达量显著降低（P < 0.05）,抗氧化酶相关基因SOD2、CAT与GPx的相对表达量显著降低（P < 0.05）;冻融组精卵结合能力均下降,但肉碱浓度为0.05 mg/mL时可得到显著改善（P < 0.05）。结果表明,添加肉碱可以改善冻融猪精子的质量、抗氧化能力、抗凋亡能力及精卵结合能力。     

Postnatal testicular development and actin appearance in the seminiferous epithelium of the Habu,Trimeresurus flavoviridis

The postnatal testicular development and actin distribution in the seminiferous epithelium were examined by light microscopy, using the testes of the Habu (Trimeresurus flavoviridis; snake) from 0-year-old to 3-year-old. At 0-year-old (about 1 month after birth), the testis was quite small in size, and the seminiferous epithelium was composed of only Sertoli cells and large spermatogonia. Actin immunoreactivity was observed in the peritubular myoid cells, but could not be detected in the seminiferous epithelium. At 1-year-old (about 10 months after birth), the testicular size increased to a great degree. In the seminiferous epithelium, spermatocytes newly appeared. Actin could still not be detected in the seminiferous epithelium. At 2-year-old (about 1 year and 10 months after birth), the testes continued to develop in size. In the seminiferous epithelium, elongate spermatids and round spermatids were frequently seen, in addition to Sertoli cells, spermatogonia and spermatocytes. Thus, active spermatogenesis was clearly recognized at this age. Moreover, the actin distribution in the seminiferous epithelium was observed at the site between Sertoli cells and spermatids, as well as that at adult stage. The immunoreactivity of actin in the peritubular myoid cells gradually increased from 0-year-old to 2-year-old. Conclusively, it seems likely that spermatogenesis in the Habu initiates at 2-year-old, accompanying with the appearance of actin in the seminiferous epithelium.     

Testicular histomorphometry and the proliferative and apoptotic activities of the seminiferous epithelium in Syrian hamster during spontaneous recrudescence after exposure to short photoperiod

Syrian hamsters are photoperiodic rodents in which reproduction, including testicular function, is stimulated by long photoperiod exposure and curtailed by exposure to a short photoperiod. The objectives of this study were to characterize the testis histomorphometrically and to determine the role of the proliferation and apoptosis phenomena in the recovery of the seminiferous epithelium during spontaneous recrudescence after exposure to short photoperiod. The study was performed using conventional light microscopy, proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase (TdT)‐mediated dUTP in situ nick end labelling staining, image analysis software, and transmission electron microscopy in three recrudescence groups: initial recrudescence (IR), advanced recrudescence (AR) and total recrudescence (TR). The results morphometrically pointed to the gradual recovery of the testicular and tubular volumes, as well as of the seminiferous epithelium. Among the IR and AR groups, the increase in testicular and tubular volumes was accompanied by an increase in tubular diameter and length, with an increase in interstitial volume. From AR to TR, there was an increase in the tubular and total volumes, but, in this case, with a gradual increase in tubular diameter. Recovery of the seminiferous epithelium was accompanied by changes in apoptosis and proliferation activities. The first decreased halfway through the process, and the second remained higher than the control levels throughout the recrudescence stage. Alterations in the spermatozoa were ultrastructurally observed, which indicated that spermiogenesis was not yet completely normal. In conclusion, spontaneous testicular recrudescence in Syrian hamster comprises two histomorphometrical phases: the first related to an increase in tubular length and diameter and interstitial volume and the second depending principally on the gradual increase in tubular diameter. The restoration of the seminiferous epithelium is due to apoptosis reaching normal values in the AR group accompanied by higher proliferative activity than that observed in the Control group.     

Perioperative outcomes after three different single extrahepatic portosystemic shunt attenuation techniques in dogs: partial ligation, complete ligation and ameroid constrictor placement

OBJECTIVE: To compare the perioperative outcomes of single extrahepatic portosystemic shunt occlusion by complete and partial silk ligation and ameroid constrictor placement in dogs. DESIGN: A retrospective analysis of 30 dogs with single congenital extrahepatic shunts. PROCEDURE: Records between 1990 and 2000 were reviewed. Patient age, breed, weight, presenting clinical signs, clinical pathology results, diagnostic imaging results, the surgery procedure performed, implant used, time taken, intra operative complications and perioperative complications were recorded. Mortality rates were calculated. RESULTS: Twenty dogs had a silk ligation procedure, 10 partially occluded and 10 completely. Ten dogs had an ameroid constrictor placement procedure. Ameroid constrictor surgery was significantly shorter in duration than silk ligation. Time for silk ligation was 91.8 +/- 35.2 minutes (median 90.0); time for ameroid constrictor placement was 71.5 +/- 12.0 (median 72.5, P = 0.049). A reduction in intraoperative complications was also noted in the ameroid constrictor surgery group. CONCLUSION: The ameroid constrictor offered a surgical occlusion procedure of single extrahepatic portosystemic shunts in dogs that was clinically as effective as silk ligation in the perioperative period, with a significantly shorter surgery time.     

Pharmacokinetics of eight anticoagulant rodenticides in mice after single oral administration

The first aim of the study was to investigate the pharmacokinetics of eight anticoagulant rodenticides (brodifacoum, bromadiolone, chlorophacinone, coumatetralyl, difenacoum, difethialone, flocoumafen and warfarin) in plasma and liver of the mouse after single oral administration. Eight groups of mice dosed orally with a different anticoagulant rodenticide in a dose equal to one‐half the lethal dose 50 (LD₅₀), were killed at various times up to 21 days after administration. The eight anticoagulant rodenticides were assayed in plasma and liver by an LC‐ESI‐MS/MS method. Depending on the compound, the limit of quantification was set at 1 or 5 ng/mL in plasma. In liver, the limit of quantification was set at 250 ng/g for coumatetralyl and warfarin and at 100 ng/g for the other compounds. The elimination half‐lives in plasma for first‐generation rodenticides were shorter than those for second‐generation rodenticides. Coumatetralyl, a first‐generation product, had a plasma elimination half‐life of 0.52 days. Brodifacoum, a second‐generation product, showed a plasma elimination half‐life of 91.7 days. The elimination half‐lives in liver varied from 15.8 days for coumatetralyl to 307.4 days for brodifacoum. The second aim of the study was to illustrate the applicability of the developed method in a clinical case of a dog suspected of rodenticide poisoning.     

五加芪粉对小鼠淋巴细胞增殖功能和抗体形成细胞的影响

为了考察五加芪粉对小鼠淋巴细胞增殖功能和抗体形成细胞的影响,为临床应用提供理论依据,设不同剂量的(60、120、240 mg·kg·d~(-1))五加芪粉组,同时设黄芪多糖粉对照组和空白对照组。连续灌胃7 d后,无菌取小鼠脾脏,制备脾淋巴细胞,采用MTT法检测小鼠淋巴细胞增殖功能,定量溶血分光光度(QHS)法测定小鼠抗体形成细胞。结果显示:五加芪粉中、高剂量组可显著(0.01P≤0.05)或极显著(P≤0.01)的提高小鼠淋巴细胞的吸光度值,具有提高机体细胞免疫力的作用;五加芪粉各给药组均显著和极显著的提高了小鼠抗体形成细胞水平(0.01P≤0.05,P≤0.01),具有提升动物体液免疫功能的作用。表明五加芪粉可同时提高机体的细胞免疫和体液免疫功能,增强机体免疫力。     

Influence of the duration of photoperiod on growth,testicular characteristics and endocrine function of boars

Crossbred boars were used to evaluate the influence of exposure to 8 or 16 hr of light daily from 75 to 175 days of age on growth rate, testicular characteristics and endocrine function. At 160 days of age, concentrations of testosterone in serum (P<.10), the areas under plotted 12 hr testosterone profiles (P<.10) and the number (P<.05) and magnitude (P<.10) of testosterone secretory spikes were increased in boars exposed to 16 hr of light compared to boars in 8 hr light, but concentrations of LH in serum were similar in boars exposed to both treatments. Treatment with GnRH resulted in similar concentrations of LH in serum for both groups of boars. Testosterone in serum after GnRH-mediated LH release was greater at .5 (P<.05) and 1.0 (P<.10) hr following GnRH in boars exposed to 16 hr of light compared to boars at 8 hr, but concentrations of testosterone were similar for both treatments from 1.5 to 4.0 hr after GnRH. Growth rate and testicular and epididymal weights and sperm reserves at 175 days of age were not significantly altered by duration of photoperiod. Boars exposed to 8 hr of light had more hair per unit area than boars exposed to 16 hr of light. We conclude that exposure of prepubertal boars to longer daily photoperiods results in increased concentrations of testosterone in serum at 160 days of age.     

Thyroid function and serum hepatic enzyme activity in dogs after phenobarbital administration

Phenobarbital is the drug of choice for control of canine epilepsy. Phenobarbital induces hepatic enzyme activity, can be hepatotoxic, and decreases serum thyroxine (T4) concentrations in some dogs. The duration of liver enzyme induction and T4 concentration decreases after discontinuation of phenobarbital is unknown. The purpose of this study was to characterize the changes in serum total T4 (TT4), free T4 (FT4), thyroid-stimulating hormone (TSH), cholesterol and albumin concentrations, and activities in serum of alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl transferase (GGT) after discontinuation of long-term phenobarbital administration in normal dogs. Twelve normal dogs were administered phenobarbital at a dosage of approximately 4.4-6.6 mg/kg PO q12h for 27 weeks. Blood was collected for analysis before and after 27 weeks of phenobarbital administration and then weekly for 10 weeks after discontinuation of the drug. The dogs were clinically normal throughout the study period. Serum ALT and ALP activity and TSH and cholesterol concentrations were significantly higher than baseline at week 27. Serum T4 and FT4 were significantly lower. Serum albumin and GGT were not changed from baseline at week 27. Changes in estimate of thyroid function (TT4, FT4, TSH) persisted for 1-4 weeks after discontinuation of phenobarbital, whereas changes in hepatic enzyme activity (ALT, ALP) and cholesterol concentration resolved in 3-5 weeks. To avoid false positive results, it is recommended that thyroid testing be performed at least 4 weeks after discontinuation of phenobarbital administration. Elevated serum activity of hepatic enzymes 6-8 weeks after discontinuation of phenobarbital may indicate hepatic disease.     

Ampicillin trihydrate in foals: serum concentrations and clearance after a single oral dose

Five foals from two to three days old were given a single oral dose of ampicillin trihydrate (20 mg/kg bodyweight [bwt]). Serum ampicillin concentrations were measured serially over a 24 h period. The study was repeated in the same foals at 16 to 21 days old. The mean peak serum ampicillin concentration at two to three days old was 5.0 micrograms/ml at 1 h after treatment; the mean peak serum concentration at 16 to 21 days old was 2.7 micrograms/ml at 2 h. The concentrations steadily declined and ampicillin was not detected in the serum from any of the foals by 24 h. Serum clearance averaged 17.7 ml/min/kg at two to three days and 35.8 ml/min/kg at 16 to 21 days.