大花杓兰的组织培养与快速繁殖

1 植物名称 大花杓兰（Cypripedium macranthum SW．）。 2 材料类别 种子。 3 培养条件 种子萌发与生长培养基：（1）Harvais（Harvais1982）＋6-BA 1．0mg·L^-1（单位下同）;（2）VWD（Van Waes和Debergh 1986）＋6-BA 1．0。原球茎分化培养基：（3）Harvais＋6-BA 0．5;（4）VWD＋6-BA 0．5。     

宁夏革螨三新种记述(蜱螨亚纲,革螨股)

记述寄螨科Parasitidae1新种,六盘山钝革螨Amblygamasus liupanshanensis Bai,Yan et Wu,sp.nov.,宽寄螨科Euryparasitidae 1新种,邓氏宽寄螨Euryparasitus tengkuofani Bai,Yan et Wei,sp.nov.和维螨科Veigaiaidae 1新种,陈氏维螨Veigaia chenbaifangi Bai,Yan et Qi,sp.nov.,标本采自宁夏六盘山自然保护区,模式标本保存于军事医学科学院微生物流行病研究所昆虫标本馆,北京。     

厌氧培养皿及厌氧培养管的临床应用——对传统的厌氧菌分离培养及鉴定方法的简化

利用我们研制的厌氧培养皿及管,共检验了39份牙髓炎及牙周炎标本,分离出厌氧菌59株,标本中厌氧菌分离的阳性率为100％。其中类杆菌28株,消化链球菌9株,消化球菌7株,韦荣氏球菌5株,真杆菌2株,梭形杆菌8株。在类杆菌中产黑素类杆菌21株,占类杆菌总数的75％。结果表明使用厌氧培养皿及管研究厌氧菌简便、有效、经济,适于基层单位应用。     

Identification of three novel proteins (SGSM1, 2, 3) which modulate small G protein (RAP and RAB)-mediated signaling pathway

We report a novel protein family consisting of three members, each of which contains RUN and TBC motifs and appears to be associated with small G protein-mediated signal transduction pathway. We named these proteins as small G protein signaling modulators (SGSM1/2/3). Northern blot analysis revealed that human SGSM2/3 are expressed ubiquitously in various tissues, whereas SGSM1 is expressed mainly in brain, heart, and testis. Mouse possessed the same protein family genes, and the in situ hybridization and immunohistochemical staining of tissue sections revealed that mouse Sgsm1/2/3 are expressed in the neurons of central nervous system, indicating the strong association of Sgsm family with neuronal function. Furthermore, endogenous Sgsm1 protein was localized in the trans-Golgi network of mouse Neuro2a cells by immunofluorescence microscopy. Expression of various cDNA constructs followed by immunoprecipitation assay revealed that human SGSM1/2/3 proteins are coprecipitated with RAP and RAB subfamily members of the small G protein superfamily. Based on these results, we postulated that the SGSM family members function as modulators of the small G protein RAP and RAB-mediated neuronal signal transduction and vesicular transportation pathways.     

Detection of insulin-releasing cells using in situ immunoblotting

Embryonic stem (ES) cells hold promise as a source for cell transplantation treatment of diseases such as type I diabetes. Further, cells releasing bioactive substances from ES cell progeny may be concentrated and purified for clinical applications. Although ES cell lines that express reporter genes have been established to isolate cells releasing bioactive substances, other difficulties must be overcome before these genetically modified cells can be used for gene therapy in human patients. Fluorescence- or magnetic-activated cell sorters are commonly used to isolate specific cells using antibodies against cell surface antigens. However, for some cells, such as insulin-producing beta cells, specific surface antigens have not yet been identified. In this study, we developed a simple and efficient method to identify and purify insulin- and alpha-fetoprotein-producing cells. A nitrocellulose membrane treated with anti-insulin or anti-alpha-fetoprotein antibodies was placed on a cell layer to trap insulin or alpha-fetoprotein released from the cells. The location of specific substance-producing cells was identified by immunostaining the membrane. The insulin-releasing cells were selectively collected from the culture dish using a cloning ring and transferred to another culture plate.     

Quantification of valproic acid and its metabolite 2-propyl-4-pentenoic acid in human plasma using HPLC-MS/MS

A specific and sensitive HPLC-MS/MS method for the quantitative determination of valproic acid (VPA) and its metabolite, 2-propyl-4-pentenoic acid in human plasma has been developed, using VPA-d15 as the internal standard. The method was based on pre-column derivatization using 4-dimethylaminobenzylamine dihydrochloride. The derivatives were separated with a gradient elution and quantified by positive electrospray ionization with multiple reaction monitoring. The assay provides routine quantification limits of 200 ng/mL for VPA and 20 ng/mL for 4-ene VPA with within- and between-day coefficients of variation of <10%. This method has been applied to the analysis of plasma samples obtained from patients treated with this drug.     

抗IFITM1单克隆抗体的制备及检测

目的：制备抗干扰素诱导的跨膜蛋白-1（interferon-induced transmembrane protein 1,IFITM1）的单克隆抗体,为检测IFITM1及进一步研究其在结肠肿瘤发生过程中的作用提供实验基础。方法：以结肠癌患者的癌组织为材料,提取总RNA,以RT—PCR扩增得到IFITM1 cDNA序列,经EcoR 和HindⅢ双酶切后,克隆入pGEX-4T-3进行原核表达并纯化得IFITM1-GST;以该融合蛋白免疫BALB／c小鼠,淋巴细胞杂交瘤法制备单克隆抗体;采用ELISA、Western-blot及免疫组织化学法以制备的抗体检测结肠癌患者结肠癌组织中的IFITM1。结果：成功构建了1FITM1核表达载体,获得了IFITM1-GST重组蛋白;制备得到了1株抗IFITM1单克隆抗体,腹水ELISA效价为1：30000,抗体亚类为IgG1,可用于ELISA、Western-blot及免疫组织化学法检测结肠癌患者结肠癌组织中的IFITM1。结论：获得了1株可用于ELISA、Westem-blot及免疫组织化学法的抗IFITM1单克隆抗体2F—1,为进一步研究IFITM1在结肠肿瘤发生过程中的作用提供了实验基础。     

Phorbol ester-potentiated liposomal transfection to monocytic PLB-985 cells

In order to improve transient gene transfer into PLB-985 cells, we treated cells with 12-tetradecanoylphorbol 13-acetate (TPA) 4 h before transfection, and increased by 540-fold the reporter activity of the firefly luciferase gene transfected by TFL-01, a cationic liposome. Dioctanolyglycerol added before TPA addition inhibited the TPA-dependent increase in activity, suggesting it to be a TPA competitor for PKC binding. H7, staurosporine, GO 6976, and H8, but not GF 109203X and forskolin, inhibited the TPA-dependent increase in reporter activity when added 8 h after TFL-01/gene addition. Forskolin and GF 109203X also inhibited the increase when added before TPA. Therefore, for the potentiation of transfection by the TPA/TFL-01 method, conventional PKC activity with significant but low protein kinase A (PKA) activity are first required, and then a novel PKC activity with significant PKA activity. TPA enhanced the uptake of FITC-labeled phosphorothioated oligonucleotides and their prolonged maintenance by cells, suggesting increased TFL-01-assisted plasmid uptake and its stabilization in TPA-treated PLB-985 cells. This method was used successfully for the sensitive analysis of the promoter function of the gp91(phox) gene, implying the method to be generally useful for promoter analyses of various genes expressed in differentiated human monocytic cells.     

中国球藓属(丛藓科)植物分布新资料

报道了球藓密疣变种在中国的新分布。该变种主要分布于欧洲、大洋洲和北美洲,在中国为首次记录。该文对其形态特征进行了详细的描述与拍照,凭证标本存放于内蒙古大学植物标本馆(HIMC)。