and laminin on adhesion and proliferation of human hepatocellular carcinoma cell

AIM:To explore the effects of PMA(phorbol-12-myristate-13-acetate, a tumor promoter, mimicking the action of diacylglycerol on PKC)and laminin on the adhesion and the proliferation of human hepatocellular carcinoma cells, and provide a new clue to liver cancer treatment.METHODS:Human hepatocellular carcinoma cell line(BEL-7402)was used to identify the endogenous laminin and protein kinase C-α(PKC-α) expression, and the effects of laminin and PMA on the adhesion and the proliferation were also investigatedin vitro.RESULTS:By the effect of exogenous laminin, human hepatocellular carcinoma cell (BEL-7402) possessed endogenous laminin expression and increased the adhesion and the proliferation, which was showed the synergistic action by the effect of PMA in combination. By the action of PMA alone, the proliferation and the PKC-α expression increased by exogenous laminin were decreased, and the adhesion and the endogenous laminin expression were increased.CONCLUSIONS:The finding suggested that the adhesion and the proliferation of human hepatocellular carcinoma cell were closely related to the effects of endogenous or exogenous laminin, which were associated with cPKC-α activity. Therefore, the application of anti-laminin antibody in combination with PKC antagonist might be a new clue to find out the therapy for liver cancer.     

Insulin induced LPL mRNA expression in THP-1 macrophage and acceleration of transferring the macrophage to foam cell

AIM:To investigate the effect of insulin on ox-LDL transferring the THP-1 cells to foam cells and influencing the LPL mRNA expression in THP-1 cells.METHODS:THP-1 cells were incubated with 50 mg/Lox-LDL and insulin at concentrations of 10 mU/L, 100 mU/L, 1 000 mU/L and 10 000 mU/L, respectively. The expression of LPL mRNA in cells was detected by RT-PCR. Lipoprotein lipase of THP-1 cells was presented by no-specific lipase staining. THP-1 cells were stained with oil red O. Accumulation of total cholesterol (TC) in THP-1 cells was determined with oxidase assay.RESULTS:In 100 mU/L、1 000 mU/L、10 000 mU/L insulin groups, LPL mRNA expression increased 2 times, the average cell perilength was longer, the percentage of positive oil red O staining cells was significant higher, the content of cholesterol in THP-1 cells was higher than in ox-LDL control (P＜0.05).CONCLUSION:Insulin accelerates transferring of THP-1 cells to foam cell with exposed to ox-LDL because LPL mRNA expression increased in the cells.     

Effects of the juice of Fructus Hippophae (JFH) on thrombocytopenia induced by cyclophosphamide in rats

AIM:To investigate the effects of the juice of fructus hippophae(JFH) on thrombocytopenia in rats.METHODS:Forty-eight wistar rats were divided into control group, model group, Yixuesheng-Jiaonang (YSJN) group and JFH group. Rats were injected intraperitoneally with cyclophosphamide (CTX, 30 mg/kg) or saline once a day for 3 days. And then, Saline, YSJN or JFH was administered (ig) once a day for 11 days. On the second, fouth, sixth and eighth day, the thrombocyte were counted, the mean platelet volume (MPV) and PDW were examined. On the eighth day, the cAMP, cGMP in platelets and platelet aggregation function were also examined.RESULTS:JFH could shorten the time of blood coagulation, increase the count of platelet, and enhance the platelet aggregation function. The experiment also showed that the JFH could decrease cGMP content in platelet.CONCLUSION:JFH could enhance blood coagulation and quality of platelet and prevent thrombocytopenia induced by cyclophosphamide in rats.     

Traumatic shock and systemic inflammation

Trauma is the No.1 cause of death in persons under the age of 45 and the No.3 cause of all deaths. Many trauma patients die of multiple organ failure (MOF), induced by systemic inflammatory response syndrome (SIRS). This paper reviews the relations of traumatic shock with systemic inflammation.     

塔里木河流域生态环境恶化的水文效应

塔里木河是我国最大的内陆河。 4条源流出山口多年平均天然径流量 2 2 4 .9× 10 8m3 (195 7～ 2 0 0 1年 )。在全球变暖的大背景下 ,塔里木河流域山区气候出现变暖增湿的趋势 ,2 0世纪 90年代径流量达 2 4 1.9× 10 8m3 ,增幅 7.6 %。由于源流区人类开发利用水资源的影响 ,使得补给塔里木河干流的水量不断减少 ,水文条件改变 ,造成干流上、中游段耗水严重 ,导致下游生态与环境急剧退化。自 2 0 0 0年以来 ,实施的从开都河—博斯腾湖向下游绿色走廊应急输水 ,已使下游生态环境开始恢复生机。     

瓢虫防卫生物碱中的氮杂大环内酯

化学防卫是昆虫防御外来侵害的方法之一。简要评述了墨西哥豆瓢虫Epilachna varivestis、南瓜瓢虫Epilachna borealis、苜蓿瓢虫Subcoccinella vigintiquatuorpunctata 3种瓢虫蛹分泌的化学防御物质——氮杂大环内酯的结构、生物活性以及生物合成的研究进展。     

The application of atomic force microscope in clinic diagnosis of erythrocytes

AIM: To investigate the application of atomic force microscope (AFM) in clinic diagnosis. METHODS: Topographic images and some parameters of large range field and microstructures of erythrocytes in the blood of normal subjects, lung cancer and myelodisplastic syndrome (MDS) patients were examined by atomic force microscope. RESULTS: Many clear topographic images of many erythrocytes, single erythrocyte, and microstructure of erythrocyte membrane surface were obtained. Many erythrocytes in lung cancer patients were found to change into echinocytes. One erythrocyte had 10-20 protuberances, most of which, with a mean width of 589. 0 nm and a length of 646. 7 nm, were on the edge of cells. The protuberances on the center of echinocytes are lodged and embedded. The erythrocytes of MDS patients were biconcave in shape. Many apertures with different diameters of tens to hundreds nanometer appeared on the surface of cell membrane. CONCLUSIONS: AFM can be widely applied in clinic pathological inspection, including quantification of cells, obtainment and comparison of many parameters (such as diameter, thickness, volume, surface, surface area/volume ratio), observation of topograph of single cell, and observation and comparison of membrane surface microstructure of cells, and so on.     

Bone marrow-derived stromal cells and cartilage tissue engineering

Articular cartilage repair is one of the most challenging issues which remains to be resolved in clinic work. Discovering of bone marrow stromal cells (BMSC) and its application in tissue engineering provide new methods for the treatment of cartilage defects. High seeding density, appropriate cytokines and three-dimensional culture play important roles in the process of inducing BMSC differentiating into chondrocytes, suitable scaffold is also essential in reconstructing cartilage in vitro by methods of tissue engineering.     

Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.     

Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.