A RAPID AND SENSITIVE METHOD FOR DETECTION OF ARYLSULFATASE ACTIVITIES

Conventional colorimetric methods for determination of arylsulfatase activities are insensitive for microassays. In this study a rapid and sensitive fluorogenic technique was used with issues of the normal levels of arylsulfatase A and B activities in leukocytes. In addition, 2 cases of genetic lysosomal storage diseases were confirmatively diagnosed, one as MLD, the other as MLS, and a third case highly suspected clinically as MLS was finally excluded by this method.     

ESTABLISHMENT OF A HUMAN T-LYMPHOMA CELL LINE（H-TL90） AND ANALYSIS OF ITS BIOLOGICAL CHARACTERISTICS

We established a human T-lymphoma cell line from the cancerous ascites of a male patient with prostate cancer which was named H-TL90. This cell line was characterized by its histological features, and by chromosomal and immunological analysis. Immunophenotypic analysis revealed that the cells expressed surface antigen CD3^- CD4^- CD7^ CD8^-. Biological analysis revealed that the cell can promote lymphocyte proliferation. This suggested that the cell line has an autosecretion function. Cytogenetic analysis revealed that H-TL90 was a hyperdlploid with 47 chromosomes and had characteristic translocation between chromosome 3 and 11, and the deletion of the long arm of chromosome 6. These results demonstrated the H-TL90 cell line can be a useful modal for the study of human T-lymphoma.     

SUPPRESSION OF MALIGNANT PHENOTYPE OF A TRANSFORMED MOUSE MAMMARY EPITHELIAL CELL LINE （11A1） BY TUMOR SUPPRESSOR GENE RB

A malignant transformed mammary epithelial cell line (11 A1) was transfected with liposome encapsulated eukaryotic expression plasrrdd pCMV-neo-RB, yielding 4 constant clones which have obvious phenotypic reversion changes, and named 11Al-R1～R4 respectively. Further experiments showed that the llA1-R1 behaved like normal epithelial cells in both morphological and hiolcgical characteristics, with decreased clonogenicity in solid argar medium as well as decreased tumorigenicity. Northern blot hybridization showed increased expression of RB gene and decreased expression of c myc gene in llA1-R1, 11A1-R2 cells compared to llA1 cells. This was an ideal phenotypic reversion model for epithelial transformed cell line and demonstrated that the RB gene can reexpress and suppress malignant phetlotype in RB inactive cells.     

EFFECTS OF RETINOIC ACID ON PROLIFERATION AND DIFFERENTIATION OF A HUMAN OVARIAN CARCINOMA CELL LINE： 3AO

Objective To observe the effects ofretinoic acid (RA) on the proliferation and differentiation of a human ovarian carcinoma cell line: 3AO cells. Methods 3AO cell proliferation was evaluated by viable cell count, percentage of cells in each cycle phase were analyzed by flow cytometric analysis, alkaline phosphatase (AKP) activity was determined as described, and CA125 expression was measured by ELISA. Results RA could inhibit the proliferation of 3AO cells accompanied with morphological changes in a dose-dependent manner. Cell cycle analysis indicated that RA inhibition of 3AO cells growth occurred through induction of G1 arrest with a concomitant reduction in the proportion of cells in S phase, AKP activity increased significantly after treatment with RA(0.1 μmol/L) for 1-5 days. Dose-response studies revealed that the AKP activity increased to a different extent as a function of RA concentrations. Furthermore, RA could suppress the expression of CA125 tumor marker in 3AO cells. Conclusion RA could markedly inhibit the proliferation and induce the differentiation of 3AO cells.     

ANDROGEN REPRESSION OF CYTOKERATIN GENE EXPRESSION DURING RAT PROSTATE DIFFERENTIATION：EVIDENCE FOR AN EPITHELIAL STEM CELL－A

ＡＮＤＲＯＧＥＮＲＥＰＲＥＳＳＩＯＮＯＦＣＹＴＯＫＥＲＡＴＩＮＧＥＮＥＥＸＰＲＥＳＳＩＯＮＤＵＲＩＮＧＲＡＴＰＲＯＳＴＡＴＥＤＩＦＦＥＲＥＮＴＩＡＴＩＯＮ：ＥＶＩＤＥＮＣＥＦＯＲＡＮＥＰＩＴＨＥＬＩＡＬＳＴＥＭＣＥＬＬ－ＡＳＳＯＣＩＡＴＥＤＭＡＲＫＥ...     

REVERSION OF MULTIDRUG RESISTANCE IN THE P-GLYCOPROTEIN POSITIVE BREAST CANCER CELL LINE（MCF-7／ADR） BY INTRODUCTION OF HAMMERHEAD RIBOZYME

A hammerhead ribozyme which slte-specifically cleaved the GUC position in codon 880 of the mdrl mRNA was designed. The target site was chosen between the two ATP binding sites, which may be impottant for the function of the P-C-p as an ATPodependent pump. A DNA sequence encoding the riboxyme gene was then incorporated into a eukaryotic expression vector (pH 0 Apt-1 neo) and transfeeted into the breast cancer cell line MCF-7/Adr, which is resistant to adrlamycin and expresses the MDR phenmype. The ribozyme was stably expressed in the cell line by the RNA dot blotting assay. The result of Northern blot assay showed that the expressed ribozyme could decrease the level of mdrl mRNA expression by 83.5%; and the expressed ribozyme could inhiblte the formation of P-glycoprotein detected by immuno-cytochemistry assay and could reduce the cell‘‘s resistance to adrimycin; this means that the resistant cells were 1 000-fold mcre resistant than the parental cell line(MCF-7), whereas those cell clones that showed ribozyme expression were only 6-fold more resistant than the parental cell line. These results show that a potentially useful tool is at hand which may inactivate MDR1 mRNA and revert the multidrug resistance phenotype.     

SPECIFIC BINDING OF HUMAN BONE MORPHOGENETIC PROTEIN （2A） WITH MOUSE OSTEOBLASTIC CELLS

Human bone morphogenetic protein 2A (hBMP2A) cDNA terminal 567 nucleotides were cloned and expressed in a phage display vector pCSM2I. Hulnata BMP2A C-terminal peptide displayed on the surface of the phage can bind specifically to the sttrface of mouse osteoblastie cell (MC3T3) membrane. ELISA assay showed a positive signal of the binding by using antibody against M13 phage gene 8 protein. After labeling with ^3HTdR,the counts of the binding groups were 3 to 10 times higher than the control groups. It suggests that the‘surface of MC3T3 cells exist the recepzor for hBMP2A.     

A RANDOMIZED MULTICENTRE COMPARATIVE STUDY ON 3 TYPES OF IUD （3 YEARS FOLLOW UP）

A randomized multicentte clinical comparative study of Stainless steel ring, VCu 200 and TCu220c has been carried out in 23 centres m 13 provinces and municipalities. A total number of 6,236 cases were recruited including 3,432 (55.04%)urban and 2,804     

人体肝癌细胞系SMMC-7721的高分辨染色体研究

应用高分辨染色体技术,分析了人体肝癌细胞系SMMC-7721染色体核型。分析表明SMMC-7721细胞系核型染色体众数为54～73条,各核型中正常染色体55～65条,异常染色体7～15条,其中可识别其结构的标记染色体13条,包括缺失、易位及等臂染色体。在每个核型中X染色体均为2N,Y染色体为N。这些染色体的数目异常与结构畸变,对认识肝细胞的转化、恶变提供了细胞遗传学基础。     

一种激活B95—8细胞增加EBV抗原表达的新方法

为获取良好的免疫原,以制备抗EBV的单克隆抗体,本文研究了用H_2O_2诱导B95—8细胞增加EBV抗原表达的实验方法及结果。实验证实用0.2 mMH_2O_2与B95—8细胞作用30分钟,再培养5天,可诱导大量EBV抗原合成。对所得结果进行统计学处理表明经H_2O_2激活的B95—8细胞,EBV抗原阳性细胞的百分率非常显著地高于未经H_20O_2处理的B95—8细胞(P<0.001)。该结果提示经H_2O_2激活的B95—8细胞可作为一种良好的免疫原,用于诊断NPC或制备抗EBV的特异性抗体。同时,鉴于H_2O_2能诱导EBV基因合成EBV抗原,而这些抗原一旦合成,瘤细胞即死亡,故H_2O_2亦可用于治疗NPC。