Test protocol of an enzyme‐linked immunosorbent assay (ELISA) for the detection of antibodies against bovine leukosis virus

Summary

In this communication the test procedure is described for an indirect enzyme‐linked immunosorbent assay (ELISA) for the detection of antibodies against bovine leukosis virus (BLV). Test sera are incubated in polystyrene microtiter plates sensitized with a partly‐purified preparation of BLV. Bovine antibodies are detected with anti‐species immunoglobulin conjugated to the enzyme horseradish peroxidase, followed by the addition of the enzyme substrate.     

急性心肌梗死患者冠状动脉支架植入术的近期疗效观察

目的 :观察急性心肌梗死患者冠状动脉支架植入术的近期疗效。方法 :4 8例急性心肌梗死 (AMI)患者 ,胸痛发作至介入治疗时间 (4 .1± 3.1)h,行急诊冠状动脉造影 ,证实冠状动脉闭塞 ,仅对梗死相关血管行支架植入。结果 :全组 4 8例术后即刻血管再通恢复心肌梗死溶栓试验为 3级者 4 3例 (93.7% )、2级者 5例 ,1例术中死于继发性心室颤动。4 8例中在梗死相关血管开通后出现心室颤动 10例 (2 0 .1% ) ,1例术后 2周后因再次心肌梗死死亡。植入术后存活的 4 6例术后随访 (8.2± 3.6 )月 ,无 1例再发生心脏事件。结论 :急性心肌梗死行冠状动脉支架术能成功挽救频死的心肌 ,能有效降低病死率。     

Effects of inhibiting myosin light chain kinase on endothelin-1 induced proliferation and apoptosis of pulmonary arterial smooth muscle cells in rats

AIM: To investigate the effect of inhibiting myosin light chain kinase(MLCK) on endothelin-1(ET-1) induced proliferation and apoptosis of rat pulmonary artery smooth muscle cells(PASMCs). METHODS: Rat PASMCs were cultured and stimulated with ET-1. The cells were randomly divided into control group, ET-1 group and ET-1+MLCK inhibitor group(ET-1+M). Western blotting, MTT assay, [³H]-TdR incorporation and flow cytometry were employed to test the expression of myosin light chain(MLC) and MLCK, cell proliferation, cell cycle and apoptotic rate of PASMCs,respectively. The phosphorylation of MLC was determined by glycerol-PAGE coupled with Western blotting. RESULTS: Compared with control group, the protein expression of MLCK and MLC phosphorylation significantly enhanced after ET-1 stimulation. ET-1 markedly induced the proliferation and decreased the percentage of apoptotic rate in the PASMCs. However, pretreatment with ML-7, a MLCK inhibitor, significantly reversed the above effects induced by ET-1. CONCLUSION: MLCK inhibitor effectively inhibits the ET-1-induced proliferation and the cell cycle progression.     

Cellular mechanisms underlying Hyperin-induced relaxation of rat basilar artery

Background and aim

Hyperin, a flavonol compound extracted from the Chinese herb Abelmoschus manihot L. Medic, is reported to exert protective actions in cerebral ischemic injury. The specific aim of the present study was to study the relaxation of Hyperin in rat isolated basilar artery and identify the underlying cellular mechanisms.

Methods

Rat isolated basilar artery segments were cannulated and perfused while being superfused with PSS solution. Vessel images were recorded by video microscopy and diameters measured. Membrane potential was recorded using glass microelectrodes to evaluate the basilar artery smooth muscle cell hyperpolarization.

Results

Perfusion of Hyperin (1 ~ 100 μM) elicited a concentration-dependent relaxation of basilar artery segments preconstricted with 0.1 μM U46619. The response was significantly inhibited by the removal of the endothelium. Hyperin also elicited marked and concentration-dependent hyperpolarization of smooth muscle cells. 30 μM nitro-L-arginine (an inhibitor of nitric oxide synthase) and indomethacin (an inhibitor of cyclooxygenase), partially inhibited Hyperin-induced relaxation and hyperpolarization leaving an attenuated, but significant, endothelium-dependent relaxation and hyperpolarization. This remaining effect was almost completely blocked by 1 mM tetraethylammonium (an inhibitor of Ca²⁺-activated K⁺ channels), or by 100 μM DL-propargylglycine, an inhibitor of cystathionine-γ-lyase (a synthase of the endogenous H₂S).

Conclusion

These findings show that Hyperin produces significant hyperpolarization in rat basilar artery smooth muscle cells and relaxation through both endothelium-dependent and endothelium-independent mechanisms. The underlying mechanisms appeared to be multi-factorial involving nitric oxide, prostacyclin, and endothelium-derived hyperpolarizing factor (EDHF). Our data further suggest that endogenous H₂S is a component of the EDHF-mediated hyperpolarization and relaxation to Hyperin.     

自体静脉或双层自体静脉（双套式）液压扩张移植修复中型动脉缺损（附23例报告）

用自体静脉液压扩张移植修复中型动脉缺损２３例（条）结果均获成功，效果良好。液压扩张后扩大了静脉移植范围，移植血管远端的器官组织远期没有缺血现象。应用双套式吻接法修复肿瘤切除所致的血管缺损，解决了以往因肿瘤侵犯较大血管而放弃手术的难题。     

环境低温对肉鸡肺小动脉壁c-jun mRNA表达的影响

研究环境低温诱发肉鸡肺动脉高压综合征(PHS)过程中肺小动脉壁c-junmRNA的表达变化,从而确定原癌基因c-jun在环境低温诱发肉鸡PHS过程中的参与作用,为肉鸡PHS发生机制的研究提供基础。160只雄性AA商品代肉鸡15日龄随机分为对照组(22±1.5)℃和低温组(11±2)℃。15~50日龄,每周每组随机取6只,做肺组织石蜡切片,应用原位杂交染色法进行c-junmRNA杂交,并结合图像分析法,测定对照组与低温组肉鸡肺小动脉壁原癌基因c-junmRNA的表达情况。1)低温组肉鸡PHS发生率(18.75%)极显著高于对照组(1.25%)(P<0.01);2)低温组肉鸡肺小动脉壁c-junmRNA的表达从22日龄开始较对照组明显增加(P<0.05),从29~50日龄较对照组极显著升高(P<0.01)。说明环境低温明显诱发了肉鸡肺小动脉壁c-junmRNA的表达,且c-junmR-NA的表达参与了环境低温诱发的肉鸡PHS的发生。     

乳腺癌术前灌注化疗与肿瘤供血的研究

本文对６例乳腺癌的肿瘤血供研究发现主要供血动脉是胸廓外侧动脉，而非肩胛下动脉和胸廓内动脉。提出行动脉灌注化疗时，不宜只行胸廓内动脉灌注，而应扩大范围，包括上述三支动脉。同时对６例乳腺癌术前灌注化疗的效果作了观察。     

新疆细毛羊肾动脉分支探讨

本研究以过氯乙烯血管铸型法和大体解剖法研究肾动脉在肾内的分支。新疆细毛羊肾属表面平滑的单乳头肾,在肾的正中额面上髓质合成一肾总乳头,但在两侧额面上肾锥体是分开的,每侧的肾锥体一般为7个。肾动脉由肾门入肾,经过2～4级分支,到相邻锥体之间为叶间动脉,每侧一般有叶间动脉8条。分其支类型有背腹型、前后型、三支型等。     

40 ℃持续热应激对肉鸡股动脉压的影响

研究40 ℃持续热应激对肉鸡动脉压的影响,探讨肉鸡热应激发生发展规律.将60只25日龄商品代雄性AA肉鸡随机分为对照组(21～24 ℃,相对湿度(50±5)%)和试验组((40±0.5)℃,相对湿度(70±5)%),分组后自由采食和饮水,对肉鸡的股动脉压进行动态检测.结果显示热暴露使肉鸡平均股动脉压((16.66±0.47),(10.81±1.59) kPa)、股动脉收缩压((18.61±0.48),(12.00±1.49) kPa)和股动脉舒张压((15.68±0.47),(10.21±1.66)kPa)极显著下降(P<0.01);热暴露10 h时使股动脉收缩压最大变化速率((51.36±20.27),(32.72±4.85) kPa·s-1)显著下降(P<0.05);热暴露后2和5 h使股动脉舒张压最大变化速率((54.97±25.91),(31.88±5.49)和(53.49±22.94),(31.92±5.56) kPa·s-1)显著下降(P<0.05);热暴露后10 h股动脉舒张压最大变化速率((58.65±18.49),(18.09±4.66) kPa·s-1)极显著下降(P<0.01).平均股动脉压与呼吸频率、体温极显著负相关(r=-0.623,r=-0.642;P<0.01).结果表明,40 ℃持续热暴露,使肉鸡的股动脉压极显著下降;股动脉压短时间内极显著下降可能为肉鸡死亡的主要原因之一.     

Effects of Rho kinase on p-Smad1 nuclear translocation in rat pulmonary artery smooth muscle cells

AIM: To explore the mechanism of bone morphogenetic protein (BMP) and Rho kinase signal pathways on the proliferation of pulmonary artery smooth muscle cells. METHODS: Pulmonary smooth muscle cells were isolated from the rat distal pulmonary artery and cultured. BMP and Rho kinase pathways were activated by BMP-2 and platelet-derived growth factor BB(PDGF-BB),respectively. Rho kinase inhibitor Y-27632 and MEK inhibitor U0126 were also used. Immunofluorescent staining was applied to observe p-Smad1 distribution across the nucleus, and the cells with positive p-Smad1 nuclear accumulation were counted and the nuclear translocation rate was calculated. The total p-Smad1 and its distribution across the nucleus were quantitatively determined by Western blotting. The cell proliferation was analyzed by CCK-8 assay. RESULTS: Exposure to BMP-2 significantly increased both the total amount of p-Smad1 and its nuclear accumulation in pulmonary smooth muscle cells. Pretreatment with PDGF-BB significantly decreased the nuclear accumulation of p-Smad1 induced by BMP-2 without decrease of total p-Smad1. However, pretreatment with Y-27632 or U0126 reversed the inhibitory effect of PDGF-BB on p-Smad1 nuclear accumulation. BMP-2 significantly inhibited the cell proliferation, but PDGF-BB blocked the effect of BMP-2 and significantly increased the cell proliferation. After pretreated with Y-27632 or U0126, the PDGF-BB-activated cell proliferation was suppressed.CONCLUSION: PDGF-BB-activated Rho kinase inhibits BMP-2-induced p-Smad1 nuclear translocation via MEK/ERK1/2, and increases pulmonary artery smooth muscle cell proliferation.