反义c-myc重组腺病毒载体的构建及其对大鼠胸腺淋巴细胞的增殖抑制作用

目的:观察反义c-myc重组腺病毒载体对大鼠胸腺淋巴细胞的增殖抑制作用.方法:构建大鼠反义及正义c-myc细菌质粒,并将所得细菌质粒与E1缺失腺病毒质粒导入293细胞系,经共转染得到正义及反义重组腺病毒载体.MTS法检测重组腺病毒载体对大鼠淋巴细胞增殖的抑制作用,RT-PCR检测重组腺病毒载体对c-myc mR-NA表达的影响.结果:反义c-myc重组腺病毒载体可抑制大鼠淋巴细胞增殖,降低淋巴细胞c-myc mRNA的表达.结论:反义c-myc重组腺病毒载体可抑制大鼠淋巴细胞增殖.     

超声破裂载基因微泡增强心肌细胞报告基因的转染与表达

目的:通过超声破裂载基因微泡介导报告基因心肌细胞转染,探讨其能否增强心肌细胞外源基因转染与表达.方法:以β-galactosidase质粒为报告基因,将其与自制氟碳气体微泡粘附,制备载基因微泡.利用诊断性超声破裂微泡进行体外心肌细胞基因转染;以磷酸钙共沉淀转染为阳性对照并将其以不同方式与超声破裂微泡技术联合应用,以期进一步增强基因转染效果.分别采用原位染色及酶学定量检测β-galactosidase表达水平,同时进行细胞活性检测.结果:超声破裂载基因氟碳气体微泡(PESDA)转染组心肌细胞β-galactosidase表达水平可达单纯质粒转染组60倍(P＜0.01).磷酸钙共沉淀转染3.67倍(P＜0.01)超声强度、微泡浓度对超声破裂介导基因转染效果有明显影响.超声破裂微泡技术与磷酸钙共沉淀联合应用可进一步提高报告基因的表达(P＜0.05),即使在磷酸钙转染后6 h,超声破裂微泡仍能明显增强报告的基因的表达(P＜0 05).结论:超声破裂微泡技术是一种高效基因转染方法,其不但能增加DNA转染,而且增强入胞后基因的表达.超声破裂微泡与其它基因转染技术联合应用能进一步增加基因转染效率.     

Different expression patterns of duplicated PHANTASTICA-like genes in Lotus japonicus suggest their divergent functions during compound leaf development

Recent studies on leaf development demonstrate that the mechanism on the adaxial-abaxial polarity pattern formation could be well conserved among the far-related species, in which PHANTASTICA （PAHN）-Iike genes play important roles. In this study, we explored the conservation and diversity on functions of PHAN-Iike genes during the compound leaf development in Lotusjaponicus, a papilionoid legume. Two PHAN-Iike genes in L. japonicus, LjPHANa and LjPHANb, were found to originate from a gene duplication event and displayed different expression patterns during compound leaf development. Two mutants, reduced leafletsl （rell） and reduced leaflets3 （rel3）, which exhibited decreased adaxial identity of leaflets and reduced leaflet initiation, were identified and investigated. The expression patterns of both LjPHANs in rel mutants were altered and correlated with abnormalities of compound leaves. Our data suggest that LjPHANa and LjPHANb play important but divergent roles in regulating adaxial-abaxial polarity of compound leaves in L. japonicus.     

两种蛙Sox基因的PCR-SSCP分析

采用PCR技术,以参考人SRI,基因HMG-box的保守区序列而设计一对特异引物,扩增了黑斑蛙和金线蛙的Sox基因。结果两种蛙的雌雄个体均扩增出217bp的基因片段,与人对照相同。对扩增产物进行SSCP分析显示,两种蛙雌雄个体间的单链迁移率相同,两种蛙之间差异较小,而与人有较大差异。测序表明,两种蛙的Sox序列之间以及与各类物种的Sox基因都有非常高的相似性,充分显示出Sox基因在系统进化上的高度保守性。     

Association between VDR ApaI Polymorphism and Hip Bone Mineral Density Can Be Modified by Body Mass Index： A Study on Postmenopausal Chinese Women

Osteoporosis is a major public health problem for old people. Genetic factors are considered to be major contributors to the pathogenesis of postmenopausal osteoporosis. The vitamin D receptor (VDR) gene is a prominent candidate gene for the regulation of postmenopausal bone mass; however, despite extensive studies, controversy remains regarding its association with postmenopausal body mineral density (BMD) variation. In this study, a total of 260 healthy postmenopausal Chinese women were genotyped at the VDR ApaI locus using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Raw hip BMD was significantly associated with VDR ApaI polymorphism with and without adjusting for age (P=0.015 and 0.040, respectively). This genetic effect can explain 3.32% of hip BMD variation. However, the significant association vanished after correcting for both age and body mass index (BMI) (P=0.169). In addition, we observed a significant association between VDR ApaI polymorphism with unadjusted BMI(P=0.042) or BMI adjusted for age (P=0.049). The raw hip BMD was also found to be significantly correlated with BMI (r=0.517, P=0.0001), with BMI explaining 26.35% of the variation of hip BMD. All of these facts prompt us to conclude that the significant association between the VDR ApaI genotype and hip BMD may be modified by BMI in postmenopausal Chinese women. Our findings may partially explain the earlier inconsistent association results concerning the VDR gene and BMD, and highlight the importance of incorporating covariates such as BMI into osteoporosis association studies.