A study on preparation of recombinant human cystatin C and its polyclonal antibody / 国际检验医学杂志

Objective To explore the preparation of recombinant human cystatin C (Cys C ) and its polyclonal antibody . Methods The gene sequences encoding Cys C were designed according to characteristics of E .coli protein-coding ,and the target gene was artificially synthesized and expressed in E .coli .The target protein was purified by affinity chromatography and was used to immunize the New Zealand white rabbits .Western blot and enzyme-linked immunosorbent assay(ELISA) were adopted to detect the recombinant human Cys C and its polyclonal antibodies in the rabbits serum .Results Purity of prepared recombinant human Cys C was more than 90% .Anti-recombinant human Cys C antibody showed good binding activities and specificity .Conclusion Prepared recombinant human Cys C and its antibody can be used in further experimental research and clinical application .

[Cytotoxicity of chymopapain combined with pingyangmycin on mouse hepatoma cell line hepa-6].

OBJECTIVE: To observe the cytotoxicity of chymopapain combined with pingyangmycin (PYM) on mouse hepatoma cell line hapa-6 in vitro. METHOD: MTT method. RESULTS: Chymopapain showed cytotoxicities to hepa-6 (IC50 is about 1200 microg/ml) and had concentration dependency. Cooperation effect was showed when three different concentration chymopapain combined with PYM during 48 hours. It accorded with the change of enzyme activity of chymopapain in saline water. Cooperation effect was showed again when adding chymopapin for the second time at 48 hours. CONCLUSION: Chymopapain has cytotoxicity on hepa-6 in vitro. Cooperation effect was showed when chymopapin was combined with PYM.

Prophylactic Effects of a HSV-2DNA Vaccine in Guinea Pigs / 中华皮肤科杂志

Objective To observe the prophylactic effects of a HSV-2gD2DNA vaccine in guinea pigs challenged with HSV-2strains.Methods Female guinea pigs were divided into3groups with10each,which was immunized intramuscularly with100?g of pc-gD plasmids(recombinant HSV-2DNA vac-cine),or with pcDNA3blank plasmids,with normal saline as control,respectively.Two booster injections were given on day7and day21.Sera were collected for virus neutralization test on day0,day28,and day56.The animals were challenged with HSV-2strain sav intravaginally,and lesions induced on the external genital skin were scored between day1and day21after challenge.Results The titer of neutralizing anti-body to HSV-2was much higher in the sera from animals immunized by pc-gD plasmids than that from ani-mals immunized by pcDNA3blank plasmids or normal saline.Furthermore,the lesion scores on external genital skin were significantly decreased in pc-gD group than those in other two groups with either primary or recurrent infections.Conclusion The constructed gD2vaccine can efficiently protect guinea pigs from genital infection and reduce recurrent infection induced by latent herpes simplex virus.

Screening for inhibitor of vascular endothelial growth factor from random peptide library / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To screen for the inhibitor of vascular endothelial growth factor (VEGF) 165 from random peptide library.</p><p><b>METHODS</b>Positive phage clones were rescued after two rounds of panning and competitive elution. Its affinity activity to KDR was monitored through ELISA, immunohistochemical method, Chicken CAM assay and MTT.</p><p><b>RESULTS</b>Five specific binding positive target molecule phage clones were obtained which were able to bind to cells whose surface had high KDR, among which, clone 3 and 13 could effectively block the vascularization of the chorioallantoic membrane of chick embryo, but they were not inhibitive on the proliferation of high KDR expression cells.</p><p><b>CONCLUSION</b>The peptides, being the inhibitors of VEGF, may be useful in the treatment of cancers.</p>

Inhibitory effect of canstatin RNA transfection on the growth of cultured rabbit vascular smooth muscle cells / 中国病理生理杂志

AIM: To investigate the effect of canstatin on cultured rabbit vascular smooth muscle cells(VSMC). METHODS: By means of cationic liposome mediated method, canstatin RNA was transferred into cultured VSMC. The proliferation quantity of VSMC were determined by the cell counting method and thymidine(-TdR) incorporation. RESULTS: Canstatin RNA could be effectively transferred into cultured primary rabbit aortic smooth muscle cells by the cationic liposome-Dosper and could markedly inhibit VSMC proliferation. CONCLUSION: Transfection of canstatin RNA could inhibit the growth of VSMC in vitro.