Influence of ovariectomy and estrogen replacement on gene expression profile of myocardium in rats

AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.     

Protective effects of ［Gly14］-humanin on cultured neural stem cells treated with β-amyloid protein in vitro

目的：研究［Gly14］-humanin对β淀粉样蛋白1-42引起的神经干细胞毒性的作用。方法：用［Gly14］-humanin和β淀粉样蛋白1-42处理神经干细胞，观察其对神经干细胞增殖、分化的影响。结果：较低浓度的［Gly14］-humanin加入有β淀粉样蛋白1-42处理的神经干细胞培养基，经过琼脂糖凝胶电泳分析及流式细胞测定DNA含量，神经干细胞显示出对β淀粉样蛋白1-42的耐受，而对照组则显示神经干细胞出现凋亡现象。高浓度的［Gly14］-humanin加入培养基，经台盼蓝拒染法计数细胞，神经干细胞死亡率明显低于对照组，对照组神经干细胞出现大量死亡。结论：［Gly14］-humanin 不但具有抑制β淀粉样蛋白1-42对神经干细胞的毒性作用，而且在低浓度的情况下，可以促进神经干细胞的增殖和分化为神经元。     

转rolA、B、C基因枳橙快繁技术

通过对转rol基因枳橙B、D、E系及对照的高接植株春梢茎段进行萌芽诱导、增殖及试管苗生根成苗的试验,摸索出适宜枳橙快繁各种培养基配方及培养条件。结果表明,以MS+6-BA1mg/L的培养基适合于芽萌发,以MS+6-BA0.5mg/L+NAA0.05mg/L的培养基适合于芽增殖,生根则以1/2MS+NAA0.5mg/L+0.2%活性炭的组合最佳,转基因各系生根率均在93.3%以上,显著高于对照生根率(66.7%)。快繁苗移栽成活率可达90%,目前生长良好。     

杨梅酸性转化酶基因cDNA分离及表达分析

杨梅果实富含蔗糖,酸性转化酶是蔗糖代谢关键酶,根据植物酸性转化酶基因保守区序列设计引物,提取杨梅叶片RNA,逆转录获得cDNA,以此为模板通过PCR技术扩增到长度为516bp的基因片段,克隆入pMD18-T载体中,命名为MrIVR1(GenBank:DQ339699)。测序及同源性检索表明,该基因推导氨基酸序列与君子兰、葡萄、草莓、胡萝卜等酸性转化酶基因氨基酸序列同源性为60%～69%。运用ClustalX软件对植物转化酶基因进行了系统树分析,结果显示,MrIVR1编码的蛋白质属于细胞壁酸性转化酶。半定量RT-PCR表达分析显示,MrIVR1基因在杨梅果实发育早期表达量最高,随着果实的发育表达量下降,在成熟果实中表达水平较低。     

柑橘新品种无子瓯柑的选育

无子瓯柑(Citrus reticulatp Blanco cv．ougan)是从普通瓯柑中选育出的芽变单株,经多子代遗传性状观察和遗传基因鉴定,证明其遗传性状稳定。果实倒卵形,果形指数1．00,平均单果质量150 g左右,果实无子,果肉橙红色,肉质细嫩,多汁,化渣,风味甜酸适口,略带苦味。果实11月中、下旬成熟,极耐贮藏,常温下可贮藏至翌年5月,且风味不变。     

牛蛙(Rana catesbeiana)蛙皮抗菌肽基因的克隆、测序及其表达

利用RT—PCR的方法从牛蛙皮肤组织中克隆到大小为270bp的片段RCABP，将其克隆到pGEM—T载体，测序获得1个新的碱基序列。将此基因克隆到原核表达载体pQE一80L，获得融合表达质粒pQE一80L／DHFR／ABP，在1％IPTG诱导下进行表达。SDS—PAGE检测表明，重组蛙皮抗菌肽蛋白的表达量占菌体总蛋白的24％，以包涵体形式存在。体外抑菌试验表明，所构建的质粒能在大肠杆菌中表达具有体外抑菌活性的蛙皮抗菌肽，该融合蛋白具有良好的应用前景。     

苏太仔猪FUT1基因M307位点多态性与F18大肠杆菌抗病相关性的体外鉴定

采用PCR-RFLP方法检测了江苏苏太断奶仔猪FUT1基因M307位点等位基因多态性分布,在所检的49头仔猪中,GG基因型个体16头,AG基因型19头,AA基因型14头。在此基础上,制备上述不同基因型个体仔猪小肠上皮细胞,分别与表达F18ab菌毛的野生型大肠杆菌、表达F18ac菌毛含fed操纵子全基因的重组大肠杆菌和V型系统表面分泌表达F18abFedF亚单位的重组大肠杆菌进行体外黏附试验和黏附抑制试验。研究结果表明:FUT1基因M307位点中GG型和AG型仔猪小肠上皮细胞均能黏附上述3种大肠杆菌,而AA型个体小肠上皮细胞则不能黏附。将上述3种大肠杆菌分别与抗F18ab菌毛高免血清、F18ac菌毛高免血清及抗F18abFedF亚单位单因子血清作用后,则失去黏附仔猪肠上皮细胞能力。上述结果对苏太猪从体外试验上证明了FUT1基因M307位点多态性与断奶仔猪腹泻和水肿病存在着直接的相关性。     

香蕉A基因组品种间遗传关系的SSR检测

应用SSR技术,对32个香蕉A基因组类型品种(系)的遗传关系进行了检测。40对SSR引物在32个品种(系)中分别扩增带数在3～15个,平均每个SSR座位可检测2.99个多态性带;引物的多态信息量(PIC)在0.00～0.88,平均0.62。依据SSR数据计算的品种间遗传距离在0.00%～34.27%,平均12.45%,大多数品种间的遗传变异非常有限,但也存在着遗传差异突出的品种:FHIA25、Yangambi KM5、Pisang Jari Buaya、Rose和皇帝蕉。依据26%的遗传距离,除了FHIA25和Pisang Jari Buaya单独化成1组外,其它30个品种可以分为2组:品种间遗传差异相对较高的组I和品种间遗传差异相对较低的组Ⅱ。Williams与引进的洪都拉斯3号、M931之间,洪都拉斯1号和洪都拉斯2号之间,高脚青芽蕉和高脚顿地雷分别没有区分开来,这可能是同物异名,也可能是同一品种未能分辨的突变体。     

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils

AIM: To explore the effect of the pretreatment of hypertonic saline (HTS) in hepatic ischemia reperfusion (I/R) injury.METHODS: The rats were divided into sham group (sham group), ischemia reperfusion group (IR group) and pretreatment of hypertonic saline group (HTS group). Partial hepatic ischemia reperfusion model was used. The rats were sacrificed at the time of 1 h, 3 h, 6 h, 12 h and 24 h after reperfusion in each group, respectively. Blood samples were obtained to examine ALT. The expression of the CD11b/CD18 (Mac-1) on the neutrophils was analyzed by flow cytometry. RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1 (ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase (MPO) in livers. The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations. RESULTS: ① HTS pretreatment decreased the level of ALT at the time points of 3 h, 6 h and 12 h after reperfusion (P<0.05). ② Mac-1 expression in HTS group was lower at 6 h and 12 h after reperfusion compared with IR group (P<0.05). ③ MPO activity in HTS group was lower at 6 h, 12 h and 24 h compared with IR group (P<0.05). ④ RT-PCR and Western blotting analysis indicated that the pretreatment of HTS inhibited the expression of ICAM-1 in livers after reperfusion. ⑤ Moderate hepatocyte swelling and few neutrophil infiltration were observed in HTS group.CONCLUSION: Pretreatment with HTS has the effect on hepatic ischemia reperfusion injury by inhibiting the expression of Mac-1 on circulating neutrophils and the expression of ICAM-1 in the liver.     

Antiproliferation effects of 188Re labeled monoclonal antibody on prostate cancer cells in vitro

AIM: To investigate the effect of 188Re labeled monoclonal antibody on prostatic specific membrane antigen 7E11C5.3,radioimmunotherapy for the treatment of human prostate cancer cell line LNCaP in vitro.METHODS: 188Re-7E11C5.3 was prepared by direct 2-mercaptoethanol reduction method.Labeling efficiency and radiochemical purity was measured by paper chromatography.Immunoreactive fraction was determined by linear extrapolation.Cytotoxicity to LNCaP cells was determined by MTT assay.RESULTS: The labeling yield of 188Re-7E11C5.3 was (93.16±2.18)%,the radiochemical purity was (95.62±0.48)%,and the immunoreactive fraction was (74.86±1.86)%.The inhibitory effect of 188Re-7E11C5.3 on cell proliferation of LNCaP cells was significantly higher than that of 188Re-mIgG or 188ReO-4.The 50% inhibitory doses (IC50) of 188Re-7E11C5.3,188Re-mIgG,and 188ReO-4 were (23.38±3.73)×107 Bq/L,(59.21±8.02)×107 Bq/L and (68.89±10.91)×107 Bq/L,respectively.CONCLUSION: 188Re-7E11C5.3 can effectively inhibit the growth of in vitro cultured prostate cancer cells and shows much potential for prostate cancer radioimmunotherapy.