氯气中毒对小鼠主要脏器水通道蛋白4表达的影响

以昆明小鼠为研究对象,探讨氯气中毒对小鼠主要脏器(心、肝、脾、肺、肾和脑)水通道蛋白4表达的影响.将40只30日龄昆明小鼠随机分为试验组和对照组,用氯气诱发中毒性肺水肿模型,剖检观察各脏器病理变化.并运用免疫组化法(SABC)检测各脏器中水通道蛋白4的表达.结果显示试验组出现肺水肿的临床症状,除肝脏外其他脏器水通道蛋白4的表达明显高于对照组(P<0.01),HE染色见水肿典型病变.结论得出氯气中毒致使小鼠主要脏器的水通道蛋白4的表达上升,这可能与水通道蛋白4的转运水的功能直接有关.     

鸭TRPV6和SLC4A4基因SNPs对蛋壳品质的影响

试验旨在探究三穗鸭TRPV6基因和SLC4A4基因SNPs突变对蛋壳品质的影响,筛选出与蛋壳品质相关的分子标记。利用PCR扩增和直接测序法相结合的方法检测鸭TRPV6基因和SLC4A4基因的SNPs,并与蛋壳品质进行关联分析。结果显示：在TRPV6基因第2内含子发现的g.81465153 C>G和g.81465176A>G突变分别对蛋壳重和蛋重有显著影响,在第3外显子发现同义突变g.81465450 T>C;3个SNPs联合产生5种单倍型和7种双倍型,位点联合对蛋壳重有显著影响。在SLC4A4基因第24外显子发现同义突变位点g.15125097 C>T,在内含子25发现g.15132523 G>A突变对蛋形指数有极显著影响,在内含子26发现g.15135079G>A突变;3个SNPs联合产生4种单倍型和7种双倍型,位点联合对蛋壳重有极显著影响。2个基因聚合后6个SNPs联合产生8种单倍型和7种双倍型,位点联合对蛋壳重和蛋重均有显著影响。表明TRPV6基因和SLC4A4基因新发现的3个SNPs联合和2个基因的6个SNPs联合均对蛋重和蛋壳重有显著效应,...     

朊蛋白在宁夏滩羊不同组织中表达的免疫组化研究

作者运用免疫组织化学技术,首次对我国一类保护物种宁夏滩羊各组织中朊蛋白(PrP)的分布进行定性、定位研究。结果显示,PrP在滩羊的大脑、脑干、小脑、心脏、肝脏、脾脏、肾脏及淋巴结均有表达,尤其在脾脏、肾脏、脑干、淋巴结中的表达量丰富,而肺脏检测结果为阴性。此研究确定了朊蛋白在滩羊各组织中的表达与分布状况,并优化了朊蛋白检测的技术条件,为羊痒病的诊断与检测提供技术支持。     

Hela细胞裸鼠模型肝脏中IL-4的表达分析

目的：通过构建Hela细胞裸鼠模型,研究白细胞介素-4(IL-4)在裸鼠肝脏中的表达情况,为研究异源移植瘤的形成提供理论基础。方法：裸鼠处死采集肝脏组织,固定后制备石蜡切片,经H&E染色观察肝脏病理结构变化,免疫组化染色观察IL-4在肝脏中的表达及分布情况。结果：H&E染色发现肝小叶结构较完整,静脉有明显的充血并在肝窦中有出血现象,肝窦内分布较多线状枯否细胞,肝小叶内有大量嗜碱性细胞聚集;免疫组化染色发现IL-4蛋白主要在肝细胞胞质中显著表达,胞核、血窦内均无表达。结论：在Hela细胞异源移植瘤的形成过程中,可能通过诱导胞质内IL-4蛋白高表达,激活相关信号通路促进肿瘤细胞的增殖分化迁移,促进肿瘤形成;肝窦中的枯否细胞极化也有可能与肿瘤形成有关。     

藏鸡与寿光鸡低氧孵化胚胎血浆中差异蛋白的筛选和鉴定

本研究采用蛋白质双向电泳和质谱鉴定技术,筛选藏鸡和寿光鸡的种蛋在低氧(13%O2)、常氧(21%O2)孵化条件下16日胚龄血浆中差异蛋白。结果显示,常氧孵化的藏鸡与寿光鸡胚胎血浆蛋白差异点仅2个,而低氧会导致8个蛋白点丰度改变;在低氧孵化条件下,藏鸡胚胎血浆比寿光鸡有较高的血浆转铁蛋白A链、载脂蛋白A-I和免疫球蛋白重链,及较低的视黄醛结合蛋白4。这4个蛋白的变化说明藏鸡胚胎对低氧耐受性高于寿光鸡,是其低氧适应性的血液学表征。本研究结果为进一步探索藏鸡胚胎低氧适应机制提供了基础。     

我国狂犬病诊断中直接快速免疫组化法的初步应用研究

目的：探讨我国在实施狂犬病诊断过程中,应用直接快速免疫组化法（DRIT）获得的诊断效果。方法：选取我国2012年12月～2014年12月收集的家犬以及患者脑组织标本共144份。针对所有标本分别选择DRIT法、DFA法（直接荧光抗体法）以及RT-PCR法（反转录-聚合酶链式反应）进行监测。对三种方法的最终检查结果进行比较,对DRIT法进行检测的敏感性以及特异性进行评价。结果：针对所有标本,选择三种方法完成检测后,最终的检测结果基本一致,DRIT法同DFA法以及RT-PCR方法进行比较,表现出相同的敏感性以及特异性。此外研究成本较低,针对技术要求较低,针对经费有限以及技术力量较弱的实验室更合适此种方法。结论：我国在进行狂犬病诊断的过程中,选择DRIT法能表现出显著的应用价值,于基层狂犬病实验室监测工作中获得广泛应用。     

雌激素对去卵巢大鼠大脑皮质和海马中Bcl-2和Bax蛋白表达的影响

利用免疫组织化学超敏SP法检测SD大鼠摘除卵巢及外源性17β-雌二醇治疗后大脑皮质和海马中Bcl-2和Bax蛋白表达的变化.结果显示,摘除双侧卵巢后,大鼠大脑皮质和海马中Bcl-2不同程度降低,而Bax的表达出现不同程度的上升,17β-雌二醇治疗后,2种蛋白的表达基本恢复正常.表明雌激素能够不同程度的上调大脑皮质和海马中Bcl-2蛋白的表达,下调Bax蛋白的表达,从而对大脑皮质及海马神经元具有保护作用.     

大黄对小鼠水通道蛋白3、4及血管活性肠肽的影响

灌胃大黄建立小鼠腹泻模型,通过测定小鼠血清中和结肠组织中水通道蛋白3(aquaporin 3,AQP3)、水通道蛋白4(aquaporin 4,AQP4)及血管活性肠肽(vasoactive intestine polypeptide,VIP)的含量变化,探究大黄致泻下的作用机制.选取40只健康昆明种小鼠,随机分为4组...     

人参三醇组皂苷对大鼠急性坐骨神经损伤修复相关蛋白表达的影响

研究人参三醇组皂苷(PTS)对大鼠坐骨神经急性损伤蛋白表达的影响。建立大鼠坐骨神经挤压损伤模型,随机分为PTS组,模型对照组及空白对照组,每组各10只大鼠;各组均经腹腔注射给药,PTS组损伤后注射PTS;模型对照组,损伤后注射同剂量生理盐水;空白对照组,不损伤坐骨神经,注射同剂量生理盐水。坐骨神经损伤术后30d,免疫荧光化学染色检测PTS对损伤侧坐骨神经神经丝蛋白(NF-M)、微管相关蛋白-2(MAP-2)、生长相关蛋白(GAP-43)表达的影响,观察PTS对损伤侧神经的影响。大鼠损伤侧坐骨神经NF-M、MAP-2、GAP-43平均光密度表达(9.66±0.99、12.43±1.44、0.344±0.040)与模型对照组(8.21±1.25、11.90±1.14、0.278±0.042)比较有极显著性差异(P0.01)。PTS可促进坐骨神经损伤大鼠NF-M,MAP-2,GAP-43的表达。     

运动对雌性SD大鼠卵巢和血清中超氧化物歧化酶的影响

为研究运动对SD大鼠卵巢和血清中超氧化物歧化酶(SOD)的影响,随机选取SD大鼠分为5组,试验组大鼠每日分别进行不同时长(0、20、40、60、80min)游泳训练。训练20周后处死大鼠,免疫组织化学SP法检测大鼠卵巢组织中SOD的表达情况,并检测血清中SOD的浓度。结果表明,40min和60min运动组血清中SOD的浓度高于对照组,差异显著(P0.05),80min运动组血清中SOD浓度显著低于对照组(P0.05);免疫组化显示,SOD主要表达于卵巢黄体的颗粒性黄体细胞中,与对照组相比,20min运动组的表达差异不显著,40min和60min运动组的表达高于对照组,差异显著(P0.05),80min运动组的表达与对照组相比差异极显著降低(P0.01)。提示,适当运动可以增加大鼠卵巢和血清中SOD的水平,而过度运动则会造成大鼠卵巢和血清中SOD水平降低。     

藏药五根散对缺血性脑损伤大鼠新皮质与海马中内皮素含量变化的影响

本文观察急性缺血性脑损伤饲喂藏药五根散后大鼠新皮质和海马中内皮素(ET)含量变化,旨在寻找藏药防治脑损伤的可行性。方法是用夹闭大鼠双侧颈总动脉制作急性脑缺血动物模型,然后给该大鼠喂服藏药,以放射免疫法测定脑组织(新皮质和海马)中ET含量,并与喂水组及正常组进行对比观察。结果表明:缺血性脑损伤后大鼠脑皮质与海马中ET含量明显升高,喂服藏药治疗组的ET含量比喂水组的明显下降(P<0.05)。表明缺血性脑损伤后大鼠新皮质及海马中的ET含量增加,而藏药可抑制ET合成使其含量下降,具有保护脑组织作用。     

Immunohistochemical Study of Steroidogenic Enzymes in the Ovary and Placenta During Pregnancy in the Dog

Using the immunohistochemical technique, we attempted to identify the source of secretion of steroid hormones between the mid- and late-terms of gestation in dogs by investigating steroid converting enzymes such as cholesterol side-chain cleavage enzyme (SCC), 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta-HSD), 17 alpha-hydroxylase/C17, 20lyase (c17), and aromatase in the ovaries and placenta. Aromatase positive cells were slightly confirmed in luteal cells in the mid-term of gestation (day 40), whereas, in the late-stage (day 50 and 60), the number of aromatase positive cells had increased. However, the oestrogen precursor (c-17 positive cells), could barely be identified in the marginal regions of the corpora lutea (CL) and completely disappeared in the late-stage of gestation. The androgen precursors, convertase SCC and 3 beta-HSD, were confirmed in all regions of the CL during the mid-stage of gestation (day 40), showing particularly strong cell reactions in the marginal region of the CL. Yet, these positive reactions of SCC and 3 beta-HSD in the marginal region of the CL disappeared in the late-stage of gestation. Moreover, it was discovered that the number of SCC and 3 beta-HSD positive cells had decreased in all regions of the CL. None of the enzymes were detected in the placenta. The above results indicated that the source of oestrogen secretion in pregnant dogs is considered to be the CL, and that, compared with the mid-stage of gestation, there was an increased number of oestrogen synthesizing cells within the CL in the late-stage. However, the biosynthetic site of oestrogen precursors from the luteal cells during the late-stage of gestation is still unknown.     

应激性抑郁症大鼠眶额叶Fos蛋白表达的变化

大脑眶额叶(OFC)是与情绪密切相关的脑区,试验主要通过强迫游泳制造应激抑郁大鼠模型,免疫组织化学方法观察模型鼠大脑OFC及其他脑区Fos蛋白表达变化,以揭示OFC区与应激性抑郁症之间的关系。结果与对照组比较,强迫游泳应激后大鼠眶额叶、海马、丘脑室旁核、杏仁内侧核等多个脑区均有Fos蛋白高表达,差异极显著(P0.01)。强迫游泳应激后OFC区Fos蛋白高表达,结果表明,OFC区参与了应激性抑郁症的发生。     

Influence of Acute Exercise on Serum Homocysteine in Horse

The effect of acute exercise on serum homocysteine (sHCy) concentration was examined in 10 horses; five Sella Italiana and five Thoroughbreds. All horses underwent standard training before the study (show jumping for Sella Italiana horses and gallop racing for Thoroughbreds). For the study, blood samples were taken at rest, immediately after exercise, and during a recovery period (30 and 60 minutes after exercise) by external jugular venipuncture. sHCy values were determined by high-performance liquid chromatography. Heart rate and blood lactate were monitored to quantify the workloads. In show jumpers, there was a significant effect of sampling time on sHCy (P < .01), which increased significantly after exercise as compared with at rest. In Thoroughbreds, no statistically significant differences were observed in sHCy over time (F_(3,12) = 0.89, P = .05). The baseline values of sHcy were higher in Thoroughbreds than in show jumpers (P < .0002). We infer that physical activity causes biochemical changes that can influence the metabolic pathway of homocysteine in show jumpers, whereas the absence of a significant effect of exercise in Thoroughbreds may reflect an adaptive response of the enzymatic antioxidant defense system.     

The Immunohistochemical Localization of Desmin and Smooth Muscle Actin in the Ovary of the African Giant Rat (Cricetomys gambianus) During the Oestrous Cycle

The aim of this study was to describe the distribution of smooth muscle actin and desmin immunopositive cells in the ovary of the giant rat. In addition, the study describes the morphological changes in the ovary of this species during the oestrous cycle. Healthy secondary and tertiary follicles dominated the ovary during pro-oestrus and oestrus. The theca externa of the tertiary follicles was immunopositive for smooth muscle actin, but immunonegative for desmin. Oestrus was also characterized by the presence of corpora haemorrhagica, which had an outer layer of smooth muscle actin immunopositive cells. Differentiating corpora lutea were observed during metoestrus. A further notable feature of the ovary during metoestrus was the presence of numerous atretic secondary and tertiary follicles. In the later stages of atresia, the follicles were infiltrated by desmin and smooth muscle actin immunopositive cells. Dioestrus was characterized by the presence of non-regressing and regressing corpora lutea. Immunostaining for smooth muscle actin was demonstrated in the enclosing layer of the corpora lutea, as well as in the tunica media of blood vessels within the corpora lutea. The results of this study have shown that morphological changes in the ovary of the giant rat during the oestrus cycle are similar to those of laboratory rodents. Furthermore, the results of the immunohistochemical study indicate that the perifollicular distribution of desmin and smooth muscle actin cells changes during follicular development and atresia.     

决明子水煎液对CCl4所致急性肝损伤大鼠血液生化指标的影响

目的:研究决明子水煎液对CCl4所致急性肝损伤大鼠血清各项生化指标的影响及对肝损伤的保护作用。方法:32只体重180～240 g的SD大鼠,随机分为正常对照组、模型对照组、决明子Ⅰ组和决明子Ⅱ组,每组8只。正常对照组和模型对照组大鼠饮用蒸馏水,决明子Ⅰ组和决明子Ⅱ组分别饮用浓度为40 g/L和80 g/L决明子水煎液,连用2周后,模型对照组、决明子Ⅰ组和决明子Ⅱ组腹腔注射CCl4(2.0 mL/kg体重),正常对照组注射等体积生理盐水,24 h后采血低速离心,取血清分别测AST、ALT、ALP、镁、磷、钾、钠、钙、胆固醇、葡萄糖、总蛋白等血液生化指标。结果:决明子水煎液(40 g/L)能极显著降低CCl4所致急性肝损伤大鼠血清中ALT、AST、血清总蛋白含量的升高(P<0.01);极显著升高CCl4所致急性肝损伤大鼠血清中胆固醇、葡萄糖、ALP和镁含量的降低(P<0.01);同时还能显著升高CCl4所致急性肝损伤大鼠血钠含量的降低(P<0.05)。结论:决明子水煎液对CCl4所致大鼠的急性肝损伤有一定保护作用,增强机体对应激的耐受力,从决明子组对各血清指标的作用来看,决明子Ⅰ组的保肝作用更优于决明子Ⅱ组。     

Fas对镉暴露致大鼠大脑皮质自噬体形成的影响

为探究死亡受体Fas在镉暴露致大鼠大脑皮质自噬体形成中的作用,将24只21日龄雄性SD大鼠随机分为4组,分别为对照组、镉组、镉与对照病毒共处理组、镉与Fas基因沉默病毒共处理组。试验期间,对照组大鼠自由饮用纯净水,病毒处理组大鼠于第1天以每只1.4×10¹¹vg的剂量通过尾静脉注射相应病毒,4周后镉染毒组大鼠自由饮用镉水(50 mg·L^-1),持续90 d。试验结束后,透射电镜观察大脑皮质中自噬体数量,Western blot检测大脑皮质细胞外信号调节激酶1/2(Erk1/2)、p-Erk1/2、自噬相关蛋白7(ATG7)、自噬相关基因(Beclin-1)、微管相关蛋白1轻链3(LC3)蛋白表达水平,免疫荧光染色检测LC3表达水平。结果显示,镉暴露增加大脑皮质中自噬体数量,极显著激活Erk1/2并上调ATG7、Beclin-1、LC3蛋白表达水平(P<0.01);Fas基因沉默抑制镉引起的自噬体数量增加,极显著抑制镉致Erk1/2激活及ATG7、Beclin-1、LC3蛋白表达水平升高(P<0.01)。结果表明,Fas通过激活Erk1/2参与镉致大鼠大脑皮质自噬体形成。     

Post‐natal Development of Perineuronal Nets in the Retrosplenial Cortex of Albino Rat

The brain extracellular matrix (ECM) has attracted growing interest due to its highly regulated spatiotemporal expression during development and maturation of central nervous system. The present study deals with the post‐natal appearance and transformation into adult distribution patterns of the ECM components related to proteoglycans (PGs) and glycoproteins (GPs) in the retrosplenial cortex (RSC) of albino rats at birth (P0), 1 week (P1), P2, P3, P4, P5, P6, P7 and P8. The differentiating PGs and GPs components of the ECM were shown to make their appearance as early as 1–2 weeks post‐natally. At this developmental stage, these components of the ECM appeared in association with some neurons and glia cells or diffusely localized at the neutrophill. Interestingly, Golgi complexes of labelled neurons were usually stained with lectin VVA or WFA, and this labelling dramatically disappeared on reaching P4. During P2–3, the pericoated neuronal cells underwent a progressive increment in number, and presented an inside‐out pattern of migration and differentiation (toward the V‐II cortical layers). On reaching P4, most of the coated neurons appeared distributed into the cortical layer IV and II. At a later stage (P5–8), the overall density and intensity of labelled neurons progressively increased and apparently reached the adult stage of development. They also displayed the usual differential labelling characteristics, after using the cationic iron colloid/lectin staining, for the first time at this juncture. The present findings indicated that the perineuronal ECM components are significantly correlated with age and suggest a possible developmental or biological significance including promotion of migration, as well as functional maturation of the retrosplenial neurons.     

3-甲基-4-硝基酚对大鼠睾丸组织的损伤作用

为从病理学角度探讨3-甲基-4-硝基酚(PNMC)的生殖毒性,采用24只SPF雄性SD大鼠随机分为4组,染毒组分为100 mg/kg体重、10 mg/kg体重和1 mg/kg体重三组,连续5 d皮下注射,对照组皮下注射等量的溶剂(PBS+0.05%tween80)。最后一次染毒后24 h,无痛处死大鼠,称取睾丸湿重并计算其指数。将已固定的睾丸组织进行石蜡切片,采用H.E和免疫组化染色的方法观察PNMC对大鼠睾丸的病理损伤及其对凋亡相关蛋白Bcl-2和Bax表达的影响。结果显示各个染毒组与对照组相比较,睾丸大体湿重和睾丸指数明显下降(P0.05或P0.01);H.E染色结果显示各试验组睾丸曲细精管生精细胞均有不同程度的空泡变性兼或坏死;免疫组织化学染色观察发现:各实验组睾丸中Bax蛋白表达量显著高于对照组(P0.01),而Bc1-2蛋白表达量却明显低于对照组(P0.01),Bc1-2/Bax也显著低于对照组(P0.01)。研究结果表明,PNMC不仅可引起雄性大鼠睾丸曲细精管生精细胞变性、坏死,还可促进生殖细胞的大量凋亡。