山羊生精上皮细胞分离研究

采用四种酶法分离二月龄关中奶山羊生精上皮细胞,A组:胶原酶Ⅳ;B组:胶原酶Ⅳ +透明质酸酶;C组:胶原酶Ⅳ+透明质酸酶+胰蛋白酶与EDTA;D组:胶原酶Ⅳ+透明质酸酶+胰蛋白酶与EDTA+DnaseI。结果二月龄羔羊曲细精管主要包含Sertoli细胞、精原细胞和管周排列的肌样细胞。每 200mg睾丸实质收获生精上皮细胞总数 (×105 )A,B,C,D组分别为 6. 91±0. 68, 6. 67±0. 80, 5. 94±0. 81, 5. 74±0. 82;细胞存活率 (% )分别为 76. 82±3. 80, 75. 96±1. 61, 94. 19±2. 89, 92. 32±1. 97;圆形细胞比率 (% )分别为 17. 54±1. 68, 16. 70±1. 46, 23. 64±1. 72, 22. 90±2. 38;细胞贴壁率(% )分别为 5. 93±0. 36, 5. 81±0. 54, 26. 94±1. 54, 25. 00±1. 74。C,D组不但组织块解离效果及细胞离散程度均较A,B好,且原代培养细胞团块数少。多重比较C,D组显著优于A,B组 (P<0.05);C组与D组差异不显著(P>0.05),但与A,B组比较细胞存活率、圆形细胞比率及贴壁率差异极显著(P<0.01)。结论认为C组是较简单有效的山羊生精上皮细胞分离酶组合。     

断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响

为了研究断奶前补饲不同直/支链淀粉比开食料对羔羊瘤胃上皮发育的影响,选取24只体况良好、胎次一致、体重相近的10日龄羔羊(湖羊),将其随机分为3组,在饲喂相同母乳的基础上,分别补饲以纯木薯(CS)、玉米(MS)和豌豆淀粉(PS)(直/支链淀粉比分别约为0.11,0.27和0.44)为唯一淀粉来源的开食料。试验期间,每天4:00-19:00将羔羊抱入补饲栏补饲不同直/支链淀粉比的开食料,并且在6:30, 10:30和15:30将羔羊抱回母羊舍哺乳1 h。羔羊自由饮水,单栏饲喂,自由采食燕麦干草。56日龄时屠宰采样,采集瘤胃组织样品制作石蜡切片进行瘤胃乳头形态测定,采集瘤胃上皮样品提取RNA测定相关基因表达。结果表明:CS组羔羊瘤胃乳头长度和表面积显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃乳头宽度显著(P=0.001)高于PS组。对瘤胃上皮各层厚度统计显示,CS和PS组羔羊瘤胃上皮角质层厚度显著(P=0.001)高于MS组;CS和MS组羔羊瘤胃上皮颗粒层厚度显著高于(P<0.001)PS组;CS组羔羊瘤胃上皮棘基层厚度和总厚度显著(P<0.001)高于MS和PS组。qRT-PCR结果显示:不同直/支链淀粉比开食料显著影响羔羊瘤胃上皮CDK2和CDK6的mRNA表达量(CS>MS>PS, P<0.001);CS组羔羊瘤胃上皮细胞cyclin A和CDK4的mRNA表达量显著(P<0.05)高于PS组,但与MS组无显著差异;CS组羔羊瘤胃上皮cyclin D1的mRNA表达量显著(P=0.012)低于PS组,但与MS组无显著差异。CS组羔羊瘤胃上皮类胰岛素生长因子-1(insulin like growth factor,IGF-1)的mRNA表达量显著(P<0.001)高于MS和PS组,CS和MS组羔羊瘤胃上皮IGF-1R的mRNA表达量显著(P=0.001)高于PS组。上述结果表明:较高支链淀粉比开食料与较高直链淀粉比开食料相比,在瘤胃中较易降解生成挥发性脂肪酸,促进瘤胃上皮IGF-1及细胞周期蛋白mRNA的表达,进而有利于断奶前羔羊瘤胃上皮的发育。研究结果对制订羔羊营养方案具有重要指导意义。     

刺参血系统的超微结构观察与功能探讨

在光学显微镜、扫描电镜和透射电镜下观察刺参血系统的组织细胞结构，探讨血系统的生理功能，结果显示：(1)刺参的血系统由复杂的管网构成，排列紧密的血管上皮细胞和上皮下连续成层的基膜，构成血系统中连续而封闭的管状结构，管腔中充满血液，血系统的管网承担了血管输送血液的功能，并通过与呼吸树的连接完成血液的气体交换。(2)血管上皮细胞游离面的细胞膜、微绒毛和吞饮小泡呈酸性磷酸酶阳性反应，表明细胞具有吸收功能。血管上皮细胞内的小泡在管壁内外的物质运输中起作用。由单层细胞和基膜构成的血管壁是血液与体腔液进行物质交换的场所。(3)血管上皮细胞富含内质网、脂滴、线粒体及分泌自噬体，具有类固醇激素分泌细胞的超微结构特点，可能与刺参体内的固醇合成有关。     

人和哺乳动物支气管动脉与肺动脉吻合现象研究进展

对人和哺乳动物支气管动脉与肺动脉吻合问题的研究进行了概述.肺动脉与支气管动脉吻合的意义在于支气管动脉的高氧血液可以进入肺动脉代偿供应肺泡组织,肺动脉的营养血液也可以进入支气管动脉代偿供应肺间质.肺动脉与支气管动脉之间血管吻合常可以发生在胸膜面、支气管壁、终末细支气管或呼吸性细支气管等部位,也可以在毛细血管上发生支气管动脉与肺动脉的吻合.根据血管吻合的长度和口径,将其分为长吻合型和短吻合型,按照血管吻合的方式分为端-端吻合和端-侧吻合.在某些病理情况下,肺动脉与支气管动脉之间的吻合支还会通过管径扩张,增加血流量来供应局部的病变组织,因此有利于防御不利因素对机体造成的损伤,为机体实施正常的呼吸功能提供保障.     

Comparison of different bronchial closure techniques following pneumonectomy in dogs

The comparison of the histologic healing and bronchopleural fistula (BPF) complications encountered with three different BS closure techniques (manual suture, stapler and manual suture plus tissue flab) after pneumonectomy in dogs was investigated for a one-month period. The dogs were separated into two groups: group I (GI) (n = 9) and group II (GII) (n = 9). Right and left pneumonectomies were performed on the animals in GI and GII, respectively. Each group was further divided into three subgroups according to BS closure technique: subgroup I (SGI) (n = 3), manual suture; subgroup II (SGII) (n = 3), stapler; and subgroup III (SGIII) (n = 3), manual suture plus tissue flab. The dogs were sacrificed after one month of observation, and the bronchial stumps were removed for histological examination. The complications observed during a one-month period following pneumonectomy in nine dogs (n = 9) were: BPF (n = 5), peri-operative cardiac arrest (n = 1), post-operative respiratory arrest (n = 1), post-operative cardiac failure (n = 1) and cardio-pulmonary failure (n = 1). Histological healing was classified as complete or incomplete healing. Histological healing and BPF complications in the subgroups were analyzed statistically. There was no significant difference in histological healing between SGI and SGIII (p = 1.00; p > 0.05), nor between SGII and SGIII (p = 1.00; p > 0.05). Similarly, no significant difference was observed between the subgroups in terms of BPF (p = 0.945; p > 0.05). The results of the statistical analysis indicated that manual suture, stapler or manual suture plus tissue flab could be alternative methods for BS closure following pneumonectomy in dogs.     

慢性阻塞性肺疾病的气道反应性与血清IL—8因子水平的研究

目的：探讨慢性阻塞性肺疾病（ＣＯＰＤ）的气道反应性ＩＬ－８因子在ＣＯＰＤ发病中的作用，方法：用组胺气道吸入激发试验观察缓解期ＣＯＰＤ，哮喘和健康人的气道反应性，同时测定气道激发前后血清ＩＬ－８水平变化。结果：ＣＯＰＤ和哮喘患者均呈气道高反应性，且ＣＯＰＤ组ＰＤ５０ＦＥＶ１与ＦＥＶ１／Ｐｒｅ％（Ｐ〈０．０２，Ｐ〈０．０５），ＩＬ－８水平与ＰＤ２０ＦＥＶ１无显著相关性，气道激发后，ＣＯＰＤ和哮喘血清Ｉ     

Epithelial response to experimentally introduced intraluminal bacteria in the avian epididymal ducts

The response of the epithelial cells of the various ducts of the avian epididymis, whose function is poorly understood, to intraluminal bacteria was evaluated by the injection of an avirulent strain of Salmonella gallinarium into the RT for 24 h. Ultrastructurally, bacteria and invading mononuclear cells were present in the lumina of the RT, proximal efferent ducts (PED) and distal efferent ducts. However, only the non-ciliated (Type I) cells of the PED epithelium ingested bacteria from the lumen. Fragments of bacteria also occurred in several intercellular spaces in the epithelium of the PED. Some mononuclear cells also contained fragments of bacteria. Neither cell death in the various epithelia nor mononuclear infiltration of the periductal tissue occurred. Therefore, in addition to the established function of absorbing most of the testicular fluid entering the epididymis, the Type I cells also appear capable of recognising and removing foreign particulate matter from the epididymal through-flow in the proximal part of the epididymis.     

单纯与联合灌注法构建猪支气管动脉立体标本

【目的】改进和优化ABS血管铸型技术。创建猪支气管动脉立体标本的铸型方法,并构建猪支气管动脉立体标本。为探明猪支气管动脉在肺内的分支与分布状态提供实验方法。【方法】用改进和优化ABS铸型技术,采用支气管动脉单纯铸型和支气管动脉与支气管树或肺动脉联合铸型的方法构建猪支气管动脉的立体标本。【结果】（1）创建了猪支气管动脉立体标本的铸型方法。①猪肺支气管动脉单纯标本制作的方法要点：先分别在胸主动脉近心端和远心端插管并结扎胸主动脉。同时,为了防止在灌入铸型剂时经食管和气管周围的血管网发生渗漏,分别在胸主动脉插管处相应的位置结扎食管前端和后端,并在气管开口处插入盲端套管并结扎。然后调整肺的位置,使其背上腹下地置于解剖盘中。经胸主动脉间接向支气管动脉中注入铸型剂。灌注时强压梯度灌入60 mL 10%和100-160 mL 15%的ABS铸型剂,当橡胶管中间部位膨大后停止灌注。在连续灌注结束后5 h内随时观察橡胶管膨大部位的大小,当压力减小时,立即灌入20%的ABS铸型剂,保证主动脉内的正压力。将灌注好的标本置于冷水中静态硬化3-4 d,再用盐酸密闭腐蚀10 d左右。然后用流水漂浮和加压冲洗,再通过摘除凝块、打枝疏密和断枝再植修整后就可以获得完整的铸型标本。②猪肺支气管动脉与支气管树联合铸型标本制作的方法要点：首先,同时插管并结扎胸主动脉、食管和气管。然后经胸主动脉间接灌注铸型剂。间隔1-2 h后再对气管树进行灌注。最后同步硬化、腐蚀、冲洗和修复支气管动脉与支气管树联合标本。③猪肺支气管动脉与肺动脉联合铸型标本制作的方法要点：支气管动脉与支气管树联合铸型的方法和支气管动脉与肺动脉联合铸型的方法相似,不同之处在于在将铸型剂注入支气管动脉的同时,前者是将铸型剂注入支气管树内,后者则是将铸型剂注入肺动脉内。在联合铸型时,根据肺的大小确定注入铸型剂的量,而经支气管动脉单纯铸型时,则根据压力大小确定灌注量。（2）构建了猪支气管动脉立体标本、猪支气管动脉与猪支气管树的联合立体标本和猪支气管动脉与肺动脉的联合标本。【结论】采用该方法获得的支气管动脉立体标本血管层次清楚、管道充盈光滑、对比度清晰,能够完整显示猪支气管动脉的起源、分支、走向、分布以及与支气管树和肺动脉的毗邻关系。该工作为猪及其它动物支气管动脉的研究奠定了基础。     

A bronchopulmonary artery fistula in a horse

A 24-year-old, Thoroughbred gelding presented with difficulty breathing for a few days and intermittent nose bleeding before dying. At necropsy, the bronchoesophageal artery and the bronchial artery that flowed into the left anterior lobe were tortuous and dilated, and it was found that dilated tortuous branches of the bronchial artery ran over the dorsal and ventral surfaces of the left anterior lobe. Histopathologically, an anastomosis between a muscular artery and an elastic artery were demonstrated, which were identified as bronchial and pulmonary arteries, respectively. Based on the gross and histopathological findings, a bronchopulmonary artery fistula was diagnosed. To the best of our knowledge, this is the first case report of a pulmonary vascular anastomosis in a horse.     

Oxidized α1-antitrypsin induces IL-8 and MCP-1 production in HBE cells

AIM:To investigate the effect of oxidized α1-antitrypsin (Ox-AT) on interleukin 8 (IL-8) and monocyte chemotactic protein 1(MCP-1) production in cultured human bronchial epithelial (HBE) cells. METHODS:Plasma native α1-antitrypsin (N-AT) was purified from human plasma by 50% and 75% ammonium sulfate fractionation followed by glutathione and anion exchange chromatography. Ox-AT was prepared by incubating N-AT (0.5 g/L) with N-chlorosuccinimide in a 25-fold molar excess to N-AT in PBS at room temperature for 30 min. HBE cells were cultured in the presence of Ox-AT (0.5 g/L) for 4 h, 10 h and 24 h, and the levels of IL-8 and MCP-1 in the supernatant were assayed using respective DuoSet kits. The effect of NF-κB inhibitor Bay11-7082 on the inflammatory cytokine release induced by Ox-AT was also evaluated. RESULTS:Ox-AT concentration-dependently and time-dependently increased the production of IL-8 and MCP-1 in HBE cells. The concentrations of IL-8 and MCP-1 in HBE cells induced by 0.5 g/L Ox-AT at 4 h, 10 h and 24 h were significantly higher than those in blank control and N-AT groups. Ox-AT increased the activity of NF-κB in a dose-dependent manner. The proinflammatory effect of by Ox-AT was inhibited by NF-κB inhibitor Bay11-7082. CONCLUSION: Ox-AT is a strong proinflammatory factor for HBE cells. The mechanism is related to NF-κB signaling pathway activation.