小鼠动情周期中卵母细胞的采集与体外成熟效果观察

本实验通过对３０只昆明白小鼠动情周期中的９９９枚卵母细胞的采集,结果表明：ＣＯＣ采集量与动情天数相关（Ｐ〈０．０１）。各天中ＣＯＣ值０ｄ最低。１ｄ这最高值,２ｄ、３ｄ下降,４ｄ上升;ＤＯ值民ＣＯＣ值相反;且ＤＯ的采集量极显著高于ＣＯＣ量（Ｐ〈０．０１）。对３３８枚卵母细胞的体外成熟结果表明。ＣＯＣ成熟率在１ｄ时达最高值。ＤＯ成熟率不受情期天数的影响。     

兔脑nNOS阳性神经元的形态、结构和分布规律

利用SABC免疫组织化学方法，对兔脑神经型一氧化氮合酶（nNOS）阳性神经元的形态、结构及分布规律进行了系统研究，结果表明：①nNOS阳性神经元呈深棕色，着色主要在胞质内，细胞核处较为浅淡；神经元胞体形态多种多样，有三角形、圆形、椭圆形、梭形等，神经元突起有一个或者数个；nNOS阳性神经纤维大多呈棕色串珠样，有些区域的阳性纤维交错分布，相互交织成网状。②家兔的各个脑区均有nNOS阳性神经元和神经纤维出现，其中在大脑皮质、小脑、丘脑下部、中脑和脑桥广泛分布，延髓分布极少。③nNOS阳性神经元，在仔兔时就已基本发育到成年兔水平，以后随着年龄增长逐渐发育成熟至衰老，到2岁龄（老年兔）时就出现了显著的衰老变化，即密度减小，表达强度减弱，第一级突起数减少，最长突起长度变短等。结果提示：nNOS阳性神经元及其催化产生的NO在家兔中枢神经系统的发育和神经调控中起重要作用。     

nNOS阳性神经元在兔大脑皮质和小脑皮质的分布

利用SABC免疫组织化学方法,对各年龄段家兔大、小脑皮质nNOS阳性神经元的分布和形态结构进行了研究。结果显示:nNOS阳性神经元主要分布在大脑皮质的Ⅱ~Ⅳ层,小脑皮质的颗粒层;nNOS阳性神经元呈深棕色,细胞质明显,细胞核着色较淡,胞体形态多样,有三角形、圆形、椭圆形、梭形等,突起有一个或者数个;阳性神经纤维大多呈棕色串珠样,有些区域的阳性纤维交错分布,相互交织成网状;在大、小脑皮质,阳性神经元密度随年龄增长而逐渐减小,分布变得稀疏,而胞体直径却逐渐增大。由此得出:家兔大脑皮质和小脑皮质内有丰富的nNOS阳性神经元分布,神经元密度随年龄增长逐渐减小,胞体直径逐渐增大;nNOS阳性神经元及其催化产生的NO在家兔中枢神经系统发育和神经调控中起重要作用。     

nNOS阳性神经元在兔大脑皮质和小脑皮质的分布

利用SABC免疫组织化学方法,对各年龄段家兔大、小脑皮质nNOS阳性神经元的分布和形态结构进行了研究。结果显示：nNOS阳性神经元主要分布在大脑皮质的Ⅱ～Ⅳ层,小脑皮质的颗粒层;nNOS阳性神经元呈深棕色,细胞质明显,细胞核着色较淡,胞体形态多样,有三角形、圆形、椭圆形、梭形等,突起有一个或者数个;阳性神经纤维大多呈棕色串珠样,有些区域的阳性纤维交错分布,相互交织成网状;在大、小脑皮质,阳性神经元密度随年龄增长而逐渐减小,分布变得稀疏,而胞体直径却逐渐增大。由此得出：家兔大脑皮质和小脑皮质内有丰富的nNOS阳性神经元分布,神经元密度随年龄增长逐渐减小,胞体直径逐渐增大;nNOS阳性神经元及其催化产生的NO在家兔中枢神经系统发育和神经调控中起重要作用。     

雌激素对奶牛输卵管上皮细胞中mPGES-1表达的影响

观察雌激素(β-雌二醇)对奶牛输卵管上皮细胞膜结合型前列腺素E2合酶-1(mPGES-1)表达的影响,并探讨β-雌二醇对奶牛输卵管上皮细胞合成分泌PGs调控的作用机制。采用胰酶消化法及机械法分离培养奶牛输卵管上皮细胞,应用荧光定量RT-PCR技术检测β-雌二醇对奶牛输卵管上皮细胞mPGES-1 mRNA表达的影响,应用Incell Western技术检测β-雌二醇对奶牛输卵管上皮细胞mPGES-1蛋白表达的影响。结果显示,与空白对照相比,β-雌二醇能够显著和极显著地促进奶牛输卵管上皮细胞中mPGES-1基因和蛋白的表达。本试验结果表明,β-雌二醇能够对奶牛输卵管上皮细胞中mPGES-1产生调节作用。     

Balb/c小鼠肺组织一氧化氮及一氧化氮合酶的动态变化

为探讨Balb/c小鼠正常生理状况下肺组织中一氧化氮自由基含量以及一氧化氮合酶活性的动态变化,采用电子自旋共振法直接测定了一氧化氮自由基含量,采用分光光度计法测定了一氧化氮合酶的活性.结果表明:在第3,6,7,12天Balb/c小鼠肺组织的一氧化氮自由基含量、一氧化氮合酶活性均无显著性差异(P＞0.05).说明生理状态下,Balb/c小鼠肺组织一氧化氮自由基的产生维持动态平衡.     

黄芪多糖对小鼠腹腔巨噬细胞iNOS基因表达及NO生成的影响

研究黄芪多糖(APS)在体内对小鼠腹腔巨噬细胞诱导型一氧化氮合酶(iNOS)基因表达及产生一氧化氮(NO)的影响。分别按5、25、50、100、200mg/kg体重,连续1周以腹腔注射方式给予小鼠APS。分离小鼠腹腔巨噬细胞,QRT-PCR检测小鼠腹腔巨噬细胞中iNOS的基因表达量;培养细胞24h,硝酸还原酶法检测上清中的NO含量,培养的单层巨噬细胞做吞噬中性红试验。结果显示,APS(100mg/kg和200mg/kg)可显著地促进巨噬细胞iNOS基因的表达和NO的产生,并显著(50、100、200mg/kg)增强巨噬细胞吞噬中性红功能,APS的作用呈浓度依赖性。体内条件下APS增强巨噬细胞功能的机制之一可能是促进巨噬细胞iNOS基因的表达进而催化产生NO。     

牦牛输卵管壶腹部与峡部的转录组学差异分析

输卵管是配子和受精卵的运输通道,在受精和早期胚胎发育中起重要作用。为深入研究牦牛输卵管不同解剖部位功能差异的分子基础,本研究对发情期母牦牛输卵管壶腹部、峡部进行了转录组测序,筛选出1395个差异表达基因,其中500个基因在壶腹部高表达,895个基因在峡部高表达。基因功能分析及通路富集结果表明,壶腹部上调差异表达基因主要与上皮细胞的完整性、细胞增殖、细胞突起、纤毛、精子运动性、受精过程有关,峡部上调差异表达基因主要与平滑肌收缩、胞外基质、解剖形态和系统发育、精子获能和储存有关。本研究从转录组层面解释了牦牛输卵管壶腹部和峡部形态结构及功能差异的原因,为进一步研究牦牛输卵管不同解剖部位的生理功能提供了基础数据。     

酮病奶牛血清中NO含量及NOS活性变化的研究

为了深入研究酮病的发生机理,选择荷斯坦奶牛作实验动物,应用酮粉法和改良式水杨醛比色法随机检测分组,Ⅰ组为10头阳性牛、Ⅱ组为10头对照牛。应用硝酸还原酶法测定血清中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性。实验结果表明对照组奶牛血清中的NO含量及NOS活性均显著大于阳性组。由此可知,酮病可导致奶牛血清中NO含量降低、NOS活性下降。     

猴头菇多糖对小鼠脾淋巴细胞体外NO分泌及iNOS mRNA表达的影响

为了研究猴头菇多糖(Hericium erinaceus polysaccharide,HEP)对小鼠脾淋巴细胞NO生成及iNOS mRNA表达的影响。采用Griess法测定HEP单独或协同ConA诱导脾淋巴细胞分泌NO含量;RT-PCR法检测脾淋巴细胞iNOS mRNA表达量。结果与细胞对照组比较,HEP在25~400μg/m L的浓度范围内能单独诱导脾淋巴细胞NO分泌及iNOS mRNA表达(P0.05);与ConA对照组比较,HEP在100~400μg/m L浓度范围内能协同Con A诱导脾淋巴细胞NO分泌及iNOS mRNA表达(P0.05)。表明HEP能单独或协同Con A诱导小鼠脾淋巴细胞分泌NO,此作用可能是通过促进i NOS mRNA表达而实现。     

去卵巢及雌激素替代治疗后家兔海马nNOS阳性神经元的形态结构及分布变化

用SABC免疫组织化学技术,观察家兔海马各区nNOS阳性神经元在去卵巢及雌激素替代治疗后的形态结构及分布变化,为雌激素类药物防治绝经后老年性痴呆症提供理论依据。结果表明,家兔海马各区都有nNOS阳性神经元分布;去卵巢后海马nNOS阳性神经元的形态结构及分布变化有区域差异性:与假手术对照组相比,在海马CA1区、CA3区、齿状回(DG)阳性神经元数量明显减少(P0.05),而在CA2区数量明显增多(P0.05)。CA1、CA3区和DG的阳性神经元胞体截面积明显变小,最长突起长度明显变短,第一级突起数变少,与假手术组有显著差异(P0.05)。CA2区阳性神经元胞体截面积明显变小(P0.05),最长突起长度、第一级突起数增多,但差异不显著(P0.05);nNOS阳性神经元的4种指标在雌激素替代治疗组与假手术组之间无显著差异(P0.05)。结果提示:雌激素可能通过影响海马nNOS的表达来影响脑的学习和记忆功能。     

肺动脉高压综合征发病过程中肉鸡血清和肺脏iNOS和cNOS活性变化

常规饲养条件喂养的大群AA肉鸡，按临床症状分为肺动脉高压组（M组），肺动脉高压腹水组（S组）和正常对照组（C组）。分别测定21、28、35和42日龄S组、M组和C组肉鸡的腹水心脏指数（AHI）、全心与体重比（WH／BW）、平均肺动脉高压（mPAP）、血清一氧化氮（NO）水平、肺脏和血清cNOS和iNOS活性。试验结果表明：（1）S组和M组全心与体重比值（TV／BW）以及S组血清NO水平均从28日龄起显著或极显著高于C组（P〈O．01或P〈0．05）；（2）S组肺组织cNOS水平显著或极显著低于C组（P〈0．01或P〈0．05）；（3）S组肺组织iNOS在21、35和42日龄显著或极显著高于C组（P〈0．01或P〈0．05）；（4）S组从28日龄起血清中cNOS和iNOS与C组差异显著或极显著（P〈0．01或P〈0．05）；（5）正常对照组肉鸡血清中iNOS活性在21、35和42日龄均显著或极显著低于血清中cNOS活性（P〈0．01或P〈0．05）。此研究结果提示cNOS和iNOS活性的变化与PHS发病机理有着密切的关系，并在其中发挥着重要的作用。     

Expression of neuronal nitric oxide synthase and renin in dysplastic kidneys of young dogs

Renin and neuronal nitric oxide synthase in the kidney control the renin-angiotensin and tubuloglomerular feedback systems. The present study investigated the expression of renin and neuronal nitric oxide synthase in the dysplastic kidneys of three young dogs. Renin-immunoreactivity, which occurs in the juxtaglomerular and tubular cells of dysplastic kidneys, did not differ from that in the normal kidneys of young dogs. Macula densa cells in the normal kidneys showed neuronal nitric oxide synthase -immunoreactivity, but those in the dysplastic kidneys showed no apparent signals. This observation may be correlated with the pathological mechanisms of renal failure in young dogs.     

Thyroid hormones alter estrous cyclicity and antioxidative status in the ovaries of rats

To expand our understanding of the roles of thyroid hormones on female reproduction, we induced hypo‐ and hyper‐T rat models to investigate the roles of thyroid hormones on estrous cyclicity, as well as the antioxidative status in the ovaries of rats. In the current study, our data show that hypothyroidism (hypo‐T) and hyperthyroidism (hyper‐T) led to significantly reduced body weights and ovarain weights and delayed vaginal opening day. For hyper‐T, thyroxine (T4), tri‐iodothyronine (T3), progesterone (P4) and follicle‐stimulating hormone (FSH) were significantly increased, while estradiol (E2) and luteinizing hormone (LH) were significantly decreased. For hypo‐T rats, serum levels of total T4 and T3, E2, P4, FSH and LH were significantly increased, while concentrations of E2 and LH were significantly decreased. For ovary morphology, the numbers of secondary and antral follicles were significantly decreased with more atretic antral follicles and less corpora lutea in both hyper‐ and hypo‐T groups. Both hyper‐T and hypo‐T treatment significantly decreased the expressions of thyroid hormone receptor α1 in the ovary. Hypo‐T significantly reduced nitric oxide (NO), total NO synthase (tNOS), inducible NOS and constitutive NOS activities, but hyper‐T increased them. For antioxidative parameters, hypo‐T and hyper‐T treatment significantly increased malondialdehyde (MDA) contents. The activities of both glutathione peroxidase (GSH‐Px) and catalase (CAT) significantly decreased in the hypo‐T group but increased in the hyper‐T group. Total superoxide dismutase (T‐SOD) activity was significantly increased in the hyper‐T group. In summary, thyroid hormones alter estrous cyclicity and antioxidative status in the ovary of the rat may act through the NOS signaling pathway.     

静脉注射L-氨基胍对肉鸡肺动脉压及其相关指标的影响

按常规饲养条件饲养AA雄性肉鸡120羽，30日龄时随机分为试验组（T组）和对照组（C组）。T组肉鸡静脉注射L-AG（40mg／kg），C组肉鸡静脉注射生理盐水。分别在注射L-AG和生理盐水1、2、4h后测定平均肺动脉压（mPAP）、诱导型一氧化氮合酶（iNOS）活性、原生型一氧化氮舍酶（cNOS）活性、一氧化氮（NO）含量、红细胞压积（PCV）、电解质浓度、pH、超氧化物歧化酶（SOD）活性、过氧化氢酶（CAT）活性和丙二醛（MDA）含量。结果显示：（1）试验组mPAP、NO含量和iNOS活性均显著或极显著低于对照组（P〈0．05或P〈0．01）；（2）PCV和Ca^2＋在给药后2h和4h与对照组差异显著（P〈0．05）；（3）K^＋浓度在给药1h和2h后显著或极显著低于时照组（P〈0．05或P〈0．01）。结果表明，L-AG通过抑制iNOS活性，引起肉鸡肺动脉压的升高，从而推测iNOS具有调节肉鸡体内mPAP的作用并与肉鸡肺动脉高压综合征发生有着密切的联系。     

一氧化氮在动物低氧适应中的生理和分子机制

近年来,国内外学者在对高原民族和高原动物低氧适应相关基因的探索中揭示了藏族的低氧适应生理机制;高含量的一氧化氮(nitric oxide,NO)舒张血管并导致高速度的血流,高速度的血流弥补了高海拔低氧。可见一氧化氮在动物低氧适应中起着重要的作用。作者对低氧条件下一氧化氮的合成、传递、生物学效应及调控表达机制进行了综述。     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.