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AIM: To investigate the contribution of angiotensin-converting enzyme inhibitor (ACEI) to the regulation of calpain system in infarcted myocardium. METHODS: Rat myocardial infarction (MI) model was established by permanent ligation of the left coronary artery. The treatment with the ACEI inhibitor rampril (1 mg·kg-1·d-1) was started 7 days prior to surgery. On day 1, 3, 7 and 14 after MI, protein levels of calpainⅠ, Ⅱ and calpastatin were determined in left ventricular free wall (LVFW), interventricular septum (IS) and right ventricule. RESULTS: CalpainⅠprotein level was increased in IS 14 d post MI, whereas the protein level of calpainⅡ was maximally increased in LVFW 3 d post MI. Rampril decreased protein up-regulation of calpainⅠ and Ⅱ, and reduced infarct size and interstitial fibrosis. Calpastatin protein expression was not affected by ACEI. CONCLUSIONS: CalpainⅠ is involved in cardiac remodelling in the late and calpainⅡ contributes to cardiac tissue damage in the early phase of MI. The heart protective effect of ACEI may be related to the inhibition of calpain system in the pathogenesis of myocardial infarction.     

烫漂及冻结对青头菌营养价值的影响

研究了在不同的烫漂温度和烫漂时间下青头菌过氧化物酶(Peroxidase，POD)活性的变化．烫漂及冻藏对青头菌过氧化物酶活性、氨基酸、矿质元素、维生素及氨基酸态氨等成分的影响结果表明：采用95～96℃烫漂温度效果最好，在4min内过氧化物酶活性降低了95．5％；烫漂及冻藏都使青头菌营养成分受到不同程度的损失建议采用一70℃以下温度不经烫漂而冻结，冻品在-26℃冰箱中贮藏12个月，其营养价值保持很好。     

红星苹果和肥城桃果实软化的酶学基础

以红星苹果穴Malusdomesticacv.Starking雪和肥城桃穴Prunuspersicacv.Feicheng雪果实为试材,结合乙烯利和1-甲基环丙烯(1-MCP)处理,分别研究了2者软化的酶学基础,并就其差别作了探讨。结果表明:红星果实软化中起主要作用的酶为内切和外切多聚半乳糖醛酸酶(endo-和exo-PG);肥城桃果实软化的前20d则以纤维素酶(EGase)为主,20d后软化以endo-PG为主。研究同时发现,脂氧合酶(LOX)在这2种果实软化的启动阶段都起了明显的促进作用。1-MCP通过对红星果实中PG、肥城桃果实贮藏前期EGase和后期endo-PG活性的抑制,以及对2者软化启动阶段LOX活性的抑制,延缓了果实硬度的下降。     

改良热硼酸法高效提取苹果果实RNA

苹果果实、尤其是成熟期的果实中多糖和其他次生物质含量高,难以提取RNA。对此,在原热硼酸法的基础上进行了改进,通过添加提取辅助剂并根据辅助剂特性调整提取步骤,高效、稳定地从苹果果实、尤其是后期果实中提取到了高质量的RNA。所得RNA的A260/A230大于2.0,A260/A280为1.8-2.1,并且RNA产量高,其中前期果实RNA产率在13.40μg/g以上,后期果实RNA产率在7.02μg/g以上,完全可以满足RT-PCR、cDNA-AFLP等分子生物学实验的要求。     

破壁蜂花粉对蛋鸡免疫功能及肠道菌群的影响

试验将槐花蜂花粉进行振动磨超微粉碎破壁,分别按1 g/kg、5 g/kg、10 g/kg三个不同剂量混到蛋鸡饲料中,从7日龄开始饲喂至50日龄,每个剂量分两个试验,试验一用新城疫Ⅳ系疫苗免疫雏鸡,利用β-微量法监测血清中的血凝抑制抗体效价的动态变化;试验二用大肠杆菌油乳苗免疫雏鸡,利用间接血凝法监测血清中大肠杆菌抗体效价的动态变化。分别定期称重捕杀取脾、胸腺、法氏囊,计算免疫器官指数,取肠道内容物计数大肠杆菌和乳酸菌,测定肠道内pH值。结果表明,不同剂量的破壁蜂花粉均能不同程度地提高病毒抗原性抗体效价和菌体抗原性抗体效价,对雏鸡免疫器官的发育以及肠道内环境、菌群的平衡也有一定影响。     

Relationship between the intracellular calcium concentration changes and left ventricular hypertrophy and function in the spontaneously hypertensive rats

AIM:To elucidate the relationship between the intracellular calcium concentration changes and left ventricular hypertrophy and function in the spontaneously hypertensive rats (SHR).METHODS:Intracellular free calcium concentrations were measured by Fura 2 methodology and left ventricular function quantitated by cardiac catheterization in 20 SHR aged 10, 22, and 34 weeks and 20 age-matched Wistar-kyoto (WKY) rats.RESULTS:(1) The systolic blood pressure(SBP), intracellular calcium concentrations and left ventricular mass / body weight index (LVM/BW) were significantly higher in all three age groups of SHR than the corresponding groups of WKY; (2) Compared with age-matched WKY groups, the peak left ventricular pressure descending rate(-dp／dt_max) decreased while left ventricular relaxation time constant (τ)increased significantly in SHR aged 22 and 34 weeks. The peak left ventricular pressure ascending rate(dp／dt_max) and the left ventricular contractility index were significantly increased only in the 34 weeks SHR; (3) Intracellular calcium concentrations showed a positive correlation with LVM/BW,SBP,-dp／dt_max and τ(r=0.47-0.83,P<0.01)and a negative correlation with dp／dt_max and the left ventricular contractility index (r=-0.46,P<0.05 and r=-0.81, P<0.01).CONCLUSION:Intracellular calcium overload is one of the potential mechanisms in the induction of left ventricular hypertrophy as well as of systolic and diastolic dysfunction.     

Protective effect of onychin on the human umbilical vein endothelial cells injured by menadione

AIM: To investigate the protective effect of onychin on the endothelial cells injured by oxidative stress. METHODS: The injured model was established by endothelial cells treated with menadione. The growth inhibitory rate of endothelial cell was determined by MTT assay; NO₂^-／NO₃^- concentration in the medium was determined by nitrate reductase assay; eNOS and caveolin-1 protein levels were determined by Western blot. RESULTS: Onychin significantly decreased the growth inhibitory rate of endothelial cells injured by menadione, increased NO₂^-／NO₃^- concentration in the medium and eNOS activity and up-regulated caveolin-1 expression. CONCLUSION: Onychin possesses a protective effect against endothelial cell injury induced by menadione via caveolin-1/eNOS pathway.     

Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.     

Biological characteristics of long passaging rat mesenchymal stem cells

AIM:To investigate multi-potential of rat bone marrow mesenchymal stem cells (rBMMSC) and mutation inclination, the rBMMSC were long passaged in vitro. METHODS:Cellular cycles of different passages were assayed by FACSan flow cytometry and karyotypes of passage 6, passage 25 and passage 45 were compared by G-binding analysis. RESULTS:The early passages and long-term passages all showed strong proliferation; passage 6, passage 25 and passage 45 all showed normal karyotype. CONCLUSION:Long-term culture and passage of rBMMSC still remains strong proliferation. With this capability, the mutation inclination is not enhanced.     

Effect of Yigu capsule drug-containing serum on apoptosis and tartrate resistant acid phosphatase secretion in rat osteoclasts

AIM:The effects of Yigu capsule on tartrate resistant acid phosphatase (TRAP) secretion and apoptosis in rat osteoclasts were investigated in order to further explore its mechanism of preventing and treating osteoporosis.METHODS:(1) Twenty-month-old Sprague-daweley rats were randomly divided into two groups(Yigu capsule group and saline group), and the drug-containing serum and control serum were prepared. (2) The newborn Sprague-daweley rat osteoclasts were cultured with different concentrations of Yigu capsule drug-containing serum. At different time point, TRAP activity was measured and the survival osteoclast was counted under reverse microscope.The percentage of osteoclast apoptosis was observed under fluorescence microscope after acridine orange staining.RESULTS:TRAP activity was lower and the percentage of osteoclast apoptosis was higher in drug-containing serum group than in control group at 24, 48 and 72 h(P<0.01), respectively, and the survival osteoclasts were less in drug-containing serum group than in control groups at 24, 48 and 72 h(P<0.01).CONCLUSIONS:These data suggest that Yigu capsule drug-containing serum induces apoptosis and inhibits TRAP activity in osteoclasts, which may be one of the mechanisms of Yigu capsule preventing and treating osteoporosis.