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。     

猪附红细胞体感染BALB/c小鼠模型的建立及相关指标检测

通过建立猪附红细胞体感染BALB/c小鼠模型,检测BALB/c小鼠外周血中血细胞的变化特征,同时检测感染小鼠血浆中红细胞表面分子CD35分子(Ⅰ型补体受体),进一步揭示附红细胞体的免疫致病机理。结果显示小鼠感染猪附红细胞体随感染时间的增加,红细胞数量呈降低趋势,白细胞数量呈升高趋势,且两者呈负相关。CD35的表达水平显著增高,说明附红细胞体感染可降低机体免疫水平。     

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。     

犬新孢子虫Nc-GFP株对BALB/c小鼠的人工感染试验

为了解犬新孢子虫Nc-GFP株的生物学特性,本试验采用Vero细胞培养的犬新孢子虫速殖子,腹腔接种BALB/c小鼠,观察小鼠的临床症状及发病情况,脱颈处死濒死期小鼠,无菌分离小鼠脑组织,接种Vero细胞进行连续培养,并提取脑组织DNA进行NcMAG1基因的PCR鉴定。结果显示:BALB/c小鼠接种犬新孢子虫Nc-GFP株速殖子后,先后出现被毛逆立、共济失调等临床症状;小鼠脑组织接种Vero细胞14 d后,在荧光显微镜下观察到绿色荧光的犬新孢子虫Nc-GFP株速殖子和包囊;小鼠脑组织经NcMAG1基因的PCR检测,扩增出1 047 bp的目的基因条带,经测序与GenBank中发表的NcMAG1(EF580924.1)核苷酸序列的同源性为99.6%。本试验成功地从BALB/c小鼠脑组织中分离出了犬新孢子虫Nc-GFP株,为Nc-GFP株生物学特性的深入研究奠定了基础。     

单侧睾丸动脉结扎／再通对小鼠生精上皮、血清抗精子抗体的影响

采用单侧睾丸动脉结扎／再通以研究其对小鼠睾丸生精功能以及血清中抗精子抗体IgG、IgM含量的影响。实验选择健康成年BALB／c雄性小鼠于显微镜下行单侧睾丸动脉结扎。再于2h、4h、6h后开通血管。ELISA法检测血清抗精子抗体IgG、IgM水平,光镜观察睾丸生精小管内变化。结果表明：与非术侧相比,术侧睾丸生精小管内生精细胞不同程度地凋亡坏死,生精上皮脱落。各手术组血清中抗精子抗体IgG、IgM含量均有明显增长（P〈0．05）。实验结论：单侧睾丸动脉结扎／再通可引起睾丸生精细胞的凋亡坏死,同时血清抗精子抗体IgG、IgM含量增加。     

SB203580对猪流感病毒诱导小鼠急性肺损伤中炎症因子的影响

为探讨SB203580对猪流感病毒(SIV)诱导小鼠急性肺损伤(ALI)中炎症因子的影响,将180只6⁸周龄SPF级BALB/c雌性小鼠,随机平均分成SIV感染组(感染组)、SIV感染+SB203580组(SB组)和对照组,于感染后2、4、6、8和14d,分别进行临床症状、肺脏病理组织学观察和肺组织中炎症因子测定。结果显示,感染组和SB组小鼠均表现明显的ALI症状,但SB组小鼠症状较轻,肺损伤程度明显低于感染组;感染组和SB组中炎症因子水平均高于对照组,SB组小鼠肺脏炎症因子水平较感染组降低。说明p38MAPK特异性抑制剂SB203580可以下调磷酸化p38MAPK的表达水平,降低炎症因子的表达,减轻ALI中肺组织的病理损伤。     

CP和NCP型牛病毒性腹泻病毒腹腔注射小鼠感染模型的建立

为了建立不同生物型牛病毒性腹泻病毒（BVDV）小鼠的急性感染模型,本研究将0.4 mL 104TCID50/mL致细胞病变型（CP型）BVDV NADL株和非致细胞病变型（NCP型）BVDV NY-1株分别经腹腔注射BALB/c小鼠,观察10 d,分别在感染后第4 d、7 d、10 d每组分别采集3只小鼠的血液、粪便后迫杀,无菌采集其心、肝、脾、肺、肠等组织样品分别进行相应检测。采集血液的血常规检测结果显示,与对照组相比,感染后第4 d、7 d和10 d,CP组和NCP组小鼠的白细胞、淋巴细胞和血小板数量均下降,在第7 d下降最明显;且感染后第7 d、10 d CP组小鼠的上述指标均低于NCP组。各组织、血液和粪便的荧光定量PCR检测结果显示,CP组小鼠在上述3个时间点血液中BVDV的载量均保持较高水平。感染后第7 d CP组和NCP组小鼠各组织中均能检测到病毒,其中血液中的病毒载量最高,对照组小鼠各组织均未检测到相应病毒。免疫组化检测感染后第7 d各组织中BVDV抗原的分布,结果显示,感染后第7 d CP组和NCP组小鼠各组织中均检测到了BVDV抗原,对照组小鼠则均未检测到相应抗原。...     

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,...     

口服猪A组轮状病毒Vp4基因工程乳酸菌对BALB/c小鼠的免疫保护作用

将轮状病毒pW425et-Vp4乳酸菌重组质粒口服免疫BALB/c小鼠,免疫后不同时间眼球采血脱臼处死,收集小鼠血清、脾细胞及肠道内容物,用ELISA方法检测血清IgG水平,用试剂盒检测肠黏膜SIgA水平,以流式细胞仪检测CD3+、CD4+、CD8+T细胞亚群和CD19+B细胞亚群的变化情况,最后进行攻毒试验。结果显示,pW425et-Vp4基因工程乳酸菌口服免疫小鼠血清IgG及肠黏膜SIgA水平较对照组有明显差异,且S/N2.0,CD4+/CD8+比值显著高于对照组,且比值大于2:1;表明猪A组轮状病毒Vp4基因工程乳酸菌重组表达质粒可以增强小鼠机体免疫保护能力。     

不同二氧化碳气腹压对犬呼吸系统的影响

充气腹是进行腹腔镜手术操作时必要步骤之一。用于充气腹的气体种类比较多,如笑气、氦气、空气等是临床上曾经采用过的气体,众多学者对以[1]     

Effects of Different Milk Samples on Sensitization in BALB/c Mice

In order to study the allergenicity of camel milk,cow milk and human milk,sixty BALB/c mice were randomly divided into β-lactoglobulin (β-lg) group (positive control group),cow milk group,camel milk group,human milk group and blank group (negative control group). Each group of mice received 1 mg/g BW samples and 0.3 μg/g cholera toxin (CT) per week,while mice in control group were received PBS and CT. After six weeks of intragastric administration,some allergic symptoms,the serum specific immunoglobulin E (IgE) and immunoglobulin G1 (IgG1) levels,plasma histamine levels and vascular permeability were detected. The results showed that the body weight of mice in camel milk,human milk and blank groups were normally increased,while that in β-lg and cow milk groups tended to slow down. The serum-specific IgE,IgG1 levels and histamine level in β-lg and cow milk groups were extremely significantly higher than blank group (P<0.01),the vascular permeability was increased,and the allergy symptoms were obvious. While the specific IgE and IgG1 levels of camel milk group were extremely significantly lower than cow milk group (P<0.01),but there were no significant difference with human milk group (P>0.05),and the allergy symptoms were mild. This study showed that the sensitization of camel milk was lower than cow milk,and similar to human milk.     

BALB/c小鼠肺组织中氧化及抗氧化系统的动态研究

为探讨BALB/c小鼠正常生理状况下肺组织中的氧化及抗氧化系统的动态变化规律,采用分光光度计法分别测定了肺组织中的黄嘌呤氧化酶（xanthine oxidase,XOD）活性、丙二醛（malondialdehyde,MDA）含量、超氧化物歧化酶（superoxide dismutase,SOD）活性变化。结果表明,在第3、6、7、12天BALB/c小鼠肺组织的XOD活性、MDA含量、SOD活性均无显著差异（P＞0.05）。说明生理状态下,BALB/c小鼠肺组织中自由基的产生与清除的酶系统之间维持动态平衡。     

Effects of Carbon Dioxide Insufflation and Body Position on Blood Gas Values in Horses Anesthetized for Laparoscopy

The objective of the study was to describe the effects of carbon dioxide pneumoperitoneum and Trendelenburg position on arterial blood gas values in horses anesthetized for laparoscopy. The study design was a prospective case series using 14 healthy adult horses anesthetized for elective laparoscopic surgery. All horses in the study were maintained under anesthesia with halothane in oxygen with intermittent positive-pressure ventilation. A pneumoperitoneum of 15 mmHg or less was achieved with carbon dioxide, and horses were tilted to a 35-degree Trendelenburg position to allow the completion of laparoscopic cryptorchidectomy (n = 13) or ovariectomy (n = 1). Heart rate, mean arterial pressure, and arterial blood gases were recorded at six time intervals throughout the procedure. Results of the study indicated a pH that decreased and partial pressure of carbon dioxide (PaCO₂) and mean arterial pressure that increased over time and differed significantly from baseline during Trendelenburg position. Partial pressure of oxygen (PaO₂) was significantly lower than baseline after assumption of Trendelenburg position and did not improve on return to normal recumbency and abdominal pressure. As body weight increased, pH and PaO₂ decreased and PaCO₂ increased. We concluded that horses placed in Trendelenburg position have changes that are transient, with the exception of PaO₂. Heavier horses have a greater change in pH, PaCO₂, and PaO₂ than lighter horses during abdominal insufflation and Trendelenburg position. The changes incurred during CO₂ abdominal insufflation and Trendelenburg position are transient, with the exception of a decreased PaO₂. Heavy horses undergoing abdominal insufflation and Trendelenburg position should be closely monitored for critical cardiopulmonary values.     

中药复合剂AFF-Ⅰ口服液对小鼠急性肝损伤血液生化指标的影响

为了研究中药复合剂AFF-Ⅰ口服液对四氯化碳(CCl4)导致小鼠急性肝损伤的保护效果,将60只昆明种小鼠随机分为正常对照、造模、阳性药物对照(联苯双酯)、中药复合剂AFF-Ⅰ口服液高、中、低剂量6组。正常对照组、造模组分别灌服等量的生理盐水,阳性药物对照组按150mg/kg灌胃联苯双酯溶液,中药复合剂AFF-Ⅰ口服液高、中、低剂量组分别按45、30、15mL/kg灌胃,2次/d,连续给药7d。末次给药后1h,正常对照组腹腔注射花生油20mL/kg,其余组腹腔注射10mL/L CCl4花生油溶液20mL/kg,16h后测定血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)含量和肝组织中超氧化物歧化酶(SOD)、丙二醛(MDA)含量。结果表明,中药复合剂AFF-Ⅰ口服液低、中、高剂量组能显著降低CCl4导致小鼠急性肝损伤血清中AST、ALT含量(P<0.01),显著降低CCl4所致急性肝损伤小鼠肝组织MDA含量,提高肝组织SOD活力(P<0.01)。研究表明,中药复合剂AFF-Ⅰ口服液对小鼠的急性肝损伤具有保护作用。     

结石通片对小鼠的急性毒性试验及其对乙二酰胺致大鼠肾结石的作用研究

[目的]研究结石通片对小鼠的急性毒性作用,对健康大鼠的利尿作用,以及对乙二酰胺致大鼠肾结石的调节作用.[方法]取ICR小鼠40只,雌、雄各半,随机分为对照组与给药组,每组20只,观察结石通片给药前后小鼠的体重变化和死亡情况;另取SD大鼠36只,雌、雄各半,分为对照组、低剂量组(给予5％结石通片溶液)和高剂量组(给予10...     

急性热应激对山羊血液生化指标及血淋巴细胞热休克蛋白70家族基因表达的影响

旨在研究急性热应激对山羊抗氧化能力、免疫功能和血淋巴细胞热休克蛋白70(HSP70)家族基因表达的影响。本试验选取5只健康、体况接近的(12±0.5)月龄波尔山羊×关中奶山羊杂交F1母羊,饲养于环控舱内(温度维持20℃,相对湿度60%),适应5d。第6天利用环控舱对5只试验羊进行38℃急性热应激处理12h,采集热应激前(0h,20℃)和热应激后2、4、8和12h试验羊血样。分别利用比色法测定血清抗氧化指标(总抗氧化能力、超氧化物歧化酶、谷胱甘肽过氧化物酶活性和丙二醛含量),ELISA法测定血清免疫指标(免疫球蛋白和细胞因子含量),实时荧光定量PCR法测定血淋巴细胞HSP70家族基因(HSPA1A、HSPA6和HSPA8)mRNA的表达量。结果显示:1)热应激时间对血清抗氧化指标有显著影响。与热应激前0h相比,血清T-AOC(P<0.05)、SOD(P<0.05)和GSH-Px(P<0.01)活性均在热应激8h后显著下降,MDA含量在热应激4h后显著增加(P<0.05)。2)热应激时间对血清免疫指标有显著影响。与热应激前0h相比,血清TNF-α、IL-1β、IFN-γ和IL-2含量分别在热应激4、8、8和4h后显著增加(P<0.05);IL-4(P<0.01)、IgG(P<0.01)、IgM(P<0.01)和IgA(P<0.05)含量分别在热应激12、4、4和4h后显著下降。3)热应激时间显著提高血淋巴细胞中HSP70家族基因(HSPA1A、HSPA6和HSPA8)的表达量。HSPA1AmRNA表达量呈先升高后下降的趋势,在热应激4h时达到峰值,各检测时间点均显著高于应激前水平(P<0.01);HSPA6mRNA表达量在热应激2h时显著升高(P<0.01),4h后恢复到应激前水平(P>0.05);HSPA8mRNA表达量在热应激4(P<0.05)、8(P<0.01)、12h(P<0.01)时显著高于应激前水平。在本试验条件下,38℃急性热应激能够抑制山羊的免疫和抗氧化功能;提高血淋巴细胞HSPA1A、HSPA6和HSPA8基因的表达量,其中HSPA1A对热应激温度和时间更敏感,可作为山羊热应激早期的分子标志物。     

腹腔注射GnIH对雄性小鼠采食、体重和血糖稳态的影响

目的 探究促性腺激素抑制激素（gonadotropin-inhibitory hormone,GnIH）对雄性小鼠采食、体重和血糖稳态的影响。方法 选取体重及日龄相近的雄性小鼠20只,随机分为2组,分别为对照组[腹腔注射生理盐水100 μL/（次·只）]和试验组[腹腔注射20 μg/100 μL GnIH,100 μL/（次·只）],每组10只,每天注射2次,连续注射21 d。观察和记录小鼠的采食情况;对小鼠的体重、空腹血糖、葡萄糖耐量、胰岛素耐量进行测定;利用实时荧光定量PCR（qPCR）方法对小鼠胰腺中胰岛素转录因子基因（NeuroD1）、胰岛素基因（Ins）、胰高血糖素基因（Gcg）、胰岛素转录调控因子基因（Pdx1）mRNA相对表达量进行检测。结果 与对照组相比,试验组小鼠平均日增重和平均日采食量极显著（P<0.01）升高;试验组小鼠空腹血糖曲线下面积极显著（P<0.01）增加,葡萄糖耐量的血糖曲线下面积极显著（P<0.01）增加,胰岛素耐量的血糖曲线下面积极显著（P<0.01）增加;试验组小鼠胰腺Gcg基因mRNA相对表达量显著（P<0.05）升高,Ins基因mRNA、Pdx1基因mRNA和NeuroD1基因mRNA相对表达量均显著（P<0.05）下降。结论 慢性腹腔注射GnIH会引起雄性小鼠采食量和体重增加,并引起机体血糖紊乱。     

Protective Effect of Stachybotrys microspora Triprenyl Phenol-7on the Deposition of IgA to the Glomerular Mesangium in Nivalenol-induced IgA Nephropathy Using BALB/c Mice

Activators of tissue proteolysis including Stachybotrys microspora triprenyl phenol (SMTP)-7 are a new class of agents that are expected to be effective for amelioration of chronic tissue destructive diseases. The present study was performed to examine whether SMTP-7 is effective for the amelioration or protection of early-stage IgA nephropathy (IgAN) induced by nivalenol (NIV) in female BALB/c mice. In Experiment 1, mice were administered NIV at 24 ppm in diet for 8 weeks, and during the NIV treatment, they were intraperitoneally injected with SMTP-7 (10 mg/kg) three times a week. In Experiment 2, mice were injected similarly with SMTP-7 during the last 4 weeks of a 16-week NIV treatment. Immunofluorescence analysis revealed an inhibitory effect of SMTP-7 on the glomerular deposition of IgA in Experiment 1; however, it was ineffective in Experiment 2. On the other hand, SMTP-7 did not affect the serum concentration of IgA in both experiments. These results suggest that SMTP-7 has a potential to decrease the progression of IgAN induced by NIV through inhibition of local accumulation of IgA in the glomerular mesangium, while it was ineffective for suppression of IgA production. On the other hand, SMTP-7 was found to be ineffective for already deposited IgA, suggesting that SMTP-7 may not be effective for ameliorating advanced IgAN.