Distribution and Expression of Protein Kinase C Interactive Protein (PKCI/HINT1) in Mouse Central Nervous System (CNS)

Protein kinase C interactive protein (PKCI; also known as histidine triad protein, HINT1) is a small intracellular protein widely expressed in tissues from both the peripheral and CNS. Although the structure of this protein is well characterized, the functional aspect and cellular distribution of the protein remain unknown, especially in CNS. To analyze the expression pattern of PKCI/HINT1 we used antibodies against either the whole recombinant protein or a peptide epitope of PKCI/HINT1. We find widespread of PKCI/HINT1 expression in the mouse CNS by Western blot and immunostaining. Our data indicates that PKCI/HINT1 is present broadly throughout the regions of CNS with relatively high abundance in olfactory system, cerebral cortex, hippocampus and part of thalamus, hypothalamus, midbrain, pons and medulla. On the cellular level, PKCI/HINT1 immunoreactivity is primarily located in neurons and neuronal processes. This study provides the anatomical evidence for the potential roles of PKCI/HINT1 in neuronal function.     

检测HIV的蛋白印迹和ELISA检测方法学比较

为了寻找一种适合大批量标本测定且具有高敏感性、高特异性的艾滋病临床实验室检测方法。比较了艾滋病检测的蛋白印迹实验和酶联免疫吸附实验两种方法,并采用不同厂家生产的不同代酶免试剂和对已确诊为阴性和阳性的标本进行比较测定。结果科华一代、二代、三代和四代酶免试剂盒的功效率分别是83．7、92．13、97．50和85.3;万泰一代、二代、三代和四代功效率分别是83．5、95．88、97．10和84．2;蛋白印迹实验的功效率是99．5。所以酶联免疫吸附实验更适合大批量标本的检测,而第三代酶免试剂具有更高的特异性和敏感性。     

Production of Transgenic Soybean Plants with Two Anti-Fungal Protein Genes Via Agrobacterium and Particle Bombardment

Utilizing either Agrobacterium-mediated transformation or particle bombardment we obtained transgenic soybean [Glycine max (L.) Merr.] plants expressing the chitinase gene (chi) and the barley ribosome-inactivating protein gene (rip). Six regenerated plants were grown to maturity and set seed. The identification of transgenic soybean plants that co-integrated the two anti-fungal protein genes was determined by polymerase chain reaction (PCR) and Southern blot analysis. Protein detection from the soybean leaves demonstrated the expression of the chitinase (CHI) and the ribosome-inactivating protein (RIP) in the six R₀ transformants. Soybean cotyledonary nodes were transformed using the bivalent plant expression vector pBRC containing chi and rip both driven by the CaMV 35S double promoter. Following vacuum (0.06 MPa) infiltration treatment of the tissue for 5 min, Agrobacterium was co-cultivated with the cotyledonary nodes for 3 d on MSB medium (MS salts and B₅ vitamins) (pH 5.2), the transformation frequency reached a maximum of 1.33 %. The chi and rip genes were present in all the transgenic plants. Co-bombardment of immature cotyledons with plasmids pBchE (encoding chi) and pARIP (encoding rip) resulted in a maximum transformation frequency of 0.52 % with a 50 % co-integration rate. Our results demonstrate efficient co-transformation of multiple genes in soybean.     

Effect of salmon melanin-concentrating hormone and mammalian gonadotrophin-releasing hormone on somatolactin release in pituitary culture of Cichlasoma dimerus

We detected a close morphological association between melanin-concentrating hormone (MCH)-immunoreactive (ir) fibers and somatolactin (SL)-ir cells in the pars intermedia of the cichlid fish Cichlasoma dimerus by double-label immunofluorescence. Male pituitaries obtained from adult C. dimerus were incubated with 0.1-10 muM salmon MCH, and the amount of SL released into the culture medium was semi-quantified by Western blot. This assay showed an increase of SL release in a dose-dependent manner (linear regression: P < 0.05). A close association of GnRH-ir fibers with SL-ir cells was also detected at the pars intermedia level. Male pituitaries were also incubated with 0.1-10 muM of mammalian GnRH, and SL release was semi-quantified by Western blot, showing an increase of released SL levels in a dose-dependent manner (linear regression: P < 0.05). In contrast, SL release was unaffected from female pituitaries incubated with salmon MCH; however, an increasing tendency was observed when mammalian GnRH was used. Hypothalamic close association of MCH-ir perikarya and GnRH-ir fibers was found by double-label immunofluorescence indicating a possible relationship between them. These results suggest that SL, like other pituitary hormones, is under hypothalamic control and is involved in diverse physiological processes including background adaptation and reproduction. This study has also shown that the in vitro culture of a single C. dimerus pituitary is a feasible method for studying the control of SL release and other pituitary hormones.     

E-cadherin 在宫颈上皮内瘤变及宫颈癌中的表达及其与高危型HPV感染的相关性

目的:研究各级宫颈上皮内瘤变(CIN)及宫颈癌组织中E-cadherin的表达及其与高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)感染的相关性探讨其在宫颈疾病发生、发展中的意义。方法:选取2008年1月至2014年12月我院收治的150例患者标本并将其分为CINⅠ级组、CINⅡ-Ⅲ级组及宫颈癌组用免疫组化法对E-cadherin的表达情况进行检测并于术前采用PCR-反向点杂交法检测患者高危型HPV感染情况所得结果:进行统计学分析。结果:(1)E-cadherin在CINⅠ级、CINⅡ-Ⅲ级及宫颈癌中的阳性表达率分别为40/50(80.0%),24/50(48.0%)、17/50(34.0%),随疾病的进展E-cadherin的表达明显减少,各组间差异有统计学意义(P0.05)。(2)高危型HPV在CINI级、CINⅡ-Ⅲ级及宫颈癌中的阳性感染率分别为21/50(42.0%)、38/50(76.0%),48/50(96.0%),各组间差异有统计学意义(P0.05)。(3)在CIN和宫颈癌中,HR-HPV阳性组中E-cadherin阳性率39.3%(42/107)低于HR-HPV阴性组中E-cadherin阳性率90.7%(39/43)(P0.05)。结论:E-cadherin的表达下降或缺失可能是HR-HPV导致宫颈癌发生、发展的机制之一。     

人肾素转基因小鼠的建立

为研究人肾素基因在体内的功能和建立其药物干预实验的动物模型,采用显微注射法,将纯化的人肾素基因导入小鼠受精卵,再培育成转基因小鼠.通过DIG DNA印迹和PCR分析,进行转基因整合检测.在出生的13只子代鼠中,得到一只转基因阳性鼠.整合率为7.7%,有效率0.3%,转基因已稳定传代.RT-PCR显示转基因阳性鼠的肾、心和肺组织中有肾素基因表达,而在肝脏与骨骼肌中则未检测到.阳性鼠血浆肾素活性较对照鼠明显升高,而肾与心脏组织的肾素活性则无明显变化.人肾素转基因小鼠可用于研究循环或组织的RAS中肾素基因的功能及有关其药物抑制实验.     

橡胶树凝集因子hevein基因及其启动子序列的分离与分析

橡胶树( Hevea brasiliensis Muell.-Arg.)凝集因子hevein是引起橡胶粒子凝集的主要因素,它是胶乳中黄色体内主要的蛋白质,具有几丁结合的功能.通过PCR技术扩增并克隆了橡胶树hevein基因共680 bp的序列.继而通过步移法分离启动子区域1 306 bp的序列,序列含典型的TATA盒和CAAT盒以及ABA效应元件的同源序列.为证实该基因在乳管中特异表达,利用Northern blot分析hevein基因在胶乳和叶片中的表达,同时,分析乙烯和ABA处理后hevein基因的表达.结果表明,hevein基因主要在胶乳中表达,乙烯和ABA对基因的表达有诱导作用.     

植物激素和抗生素调节的转基因火炬松愈伤组织的生长和分化

包含质粒载体pBI121的农杆菌菌株LBA4404被用于转化火炬松的成熟合子胚。质粒载体pBI121含有胭脂碱合成酶基因的启动子驱动的新霉素磷酸转移酶基因（npt Ⅱ)和花椰菜花叶病毒的35S启动子驱动的GUS基因。器官发生的转基因愈伤组织和转基因的再生植株已经获得,并经GUS组织化学染色、聚合酶链式反应和Southern杂交分析证实。植物激素（BA/IBA）和抗生素对转基因愈伤组织的生长和分化的影响被研究。500mg/L羧苄青霉素和2mg/LBA、0.5mg/L IBA的组合导致转基因愈伤组织的生长增加54.2%,分化增加45.7%。500mg/L Claforan和2mg/L BA、0.5mg/L IBA的组合导致转基因愈伤组织的生长增加40.8%,分化增加38.7%。高浓度的植物激素和抗生素降低了转基因愈伤组织的生长和分化。实验结果有助于建立一个高效的农杆菌介导的火炬松遗传转化系统,也有助于未来针叶树的遗传转化研究。