To investigate the inhibitory effects of siRNA expression vector on proto-oncogene Pokemon expression in SW480 cells and to provide experimental basis for further research about the biological function of Pokemon.siRNA expression vectors were constructed to express a short hairpin RNA against Pokemon.The recombinants were transfected into SW480 cells with liposome.Cellular morphology and transfection efficiency were observed.The expression of Pokemon were checked by real-time fluorescence quantitative PCR and Western blot.MTT assay was used to detect the effect of siRNA on the growth of SW480 cells and Fluorescence Activated Cell Sorter was applied to study cell apoptosis of SW480 cells.The transfection efficiency was about 36% and the cellular morphology changed greatly at 48h after transfection.siRNA expression vectors could specifically reduce the expression of Pokemon mRNA and protein in SW480 cells.Compared with negative control,the inhibition ratio of Pokemon mRNA expression was 34.2% and 67.7%,in 24th hour and 48th hour,the inhibition ratio of Pokemon protein was 48.3% and 73.6%,in 48th and 72th hour,respectively.Transfection with Pokemon siRNA expression vectors could inhibit cell growth and induce cell apoptosis of SW480 cells.siRNA expression vectors were successfully established that could effectively inhibit the expression of Pokemon in SW480 cells.Pokemon gene silencing could decrease growth and increase apoptosis of SW480 cells.

Dermatitis, Occupational ; diagnosis ; Diagnostic Errors ; Female ; Humans ; Trichloroethylene ; adverse effects ; Young Adult

Objective To explore a better method to repair cervical cicatricle contracture deformity.Methods The cervicai cicatrical contraction deformity was repaird by the cervico-shoulder island skin flap with blood supply crossing and abundant anastomotic branches of the neck-shoulder blood vessels. After removing the cervical scar, the flaps were designed according to the size of the wound to be repaired, and the clinical anatomy of crossing and abundant anastomotic branches of the neck-shoulder blood vessels. The secondary wound of donor site was closed directly if the donor region was small, or closed by skin graft. Results Sixteen patients in this clinic were operated with this method, and venous return dysfunction occurred in 1 case, but blood circulation was improved after treatment. All the flaps survived with good appearance and texture. The extensive flap was created and transferred to the anterior neck by one stage without pre-expansion which met some patients'requirement of repairing cervical cicatricle contracture deformity. Conclusion The cervico-shoulder island skin flap, with abundant blood supply, is a new method to repair cervical cicatricle contracture deformity.

Objective To explore the cosmetic effect of oncoplastic volume displacement in breastconserving surgery.Methods 43 cases of breast cancer in stages I and Ⅱ with pathological diagnosis by core needle biopsy were operated by extended resection of tumor combined with axillary lymph nodes dissection.Among the patients,wound cavity was repaired by direct skin suture without suture of mammary gland deletion in 22 cases,and another 21 cases were repaired by oncoplastic volume displacement technique in which skin suture followed by freeing remaining breast tissue flap and translocation to repair the mammary defect.All of them were treated with three-dimension conformal radiotherapy,standard chemotherapy and endocrine therapy after operation.3 years after partial removal of breast,a retrospective historical control study had been fulfilled to analyze breast cosmetic results between two groups.Results In direct suture group,the cosmetology score was 4.3 points and fine rate was 40.9% (9/22).In breast reconstruction group.the score was 4.8 points and fine rate was 71.4% (15/21).The breast cosmetic effect in breast reconstruction group was significantly better than that in direct suture group (P<0.01).Conclusion It will get better breast cosmetic results after operation to utilize oncoplastic volume displacement suture technique in breast-conserving surgery.

Objective To construct the competency model of excellent and common clinical medicine undergraduates according to the competency theory . Methods Literature search was used to screen competency model. Behavioral event interview was used to interview 32 excellent clinical experts. Open questionnaire survey was conducted to 100 clinical medical undergraduates. We compiled the primary competency dictionary based on the above results. To determine the final competency dictionary, Delphi method of 50 experts was used to determine the final competency dictionary. According to the competency dictionary, a questionnaire survey was made to test the excellent and ordinary undergraduate students. Results The 38 features of behavioral event interview and 42 features of open questionnaire survey were combined to 51 features, which were used as the first round of expert questionnaire. Two rounds of expert consultation reached a high degree of coordination. 4 first level indicators: knowledge and skills, social role+values+service characteristics, self-concept and consciousness, personality quality and motivation and 36 features were established as final compentency model. 80 excellent clinical medical undergraduates got score of 143.65 ±23.53, while 100 ordinary undergraduates got score of 133.55 ±35.31 (t=2.29,P=0.023), which indicated the competency model could distinguish excellent and ordinary clinical under-graduates . Conclusion This competency model of clinical undergraduate students is reliable and practical, which can be used in future clinical undergraduate training and evaluation.

Objective To explore the importance effect of circulating endothelial cells(CEC)in the pathogenesis of Henoch-Schonlein purpura(HSP)by determining the level of CEC in children with HSP,and to explore the relationship between CEC and therapeutic effect of HSP and the relationship between CEC and Henoch-Schonlein purpura nephritis(HSPN).Methods Sixty-six inpatients with HSP and 30 children as healthy control group were selected in the First Affiliated Hospital of Zhengzhou University from May to Dec.2008.The blood of all children,including the children in healthy control group,were sampled 1 mL in the early morning.And the CEC of all children were measured by flow cytometry method.The variation of CEC was analyzed in patients with HSP at acute stage and in healthy control group,in patients with HSP and in patients with HSPN,and in patients with HSP without renal damage at recovery stage and in patients with HSPN at recovery stage.SPSS 13.0 statistical software was used to analyze the data.Results CEC in patients with HSP at acute stage[(47.934 5?16.902 6)%]was significantly higher than that in healthy control group[(37.436 7?10.200 7)%](t=2.900 P0.05).CEC in patients with HSPN at recovery stage[(50.258 8?12.824 2)%]was significantly higher than that in patients with HSP without renal damage at recovery stage [(40.864 0?8.165 2)%](t=2.906 P

No abstract available.
Drug Therapy* ; Ovarian Neoplasms*

Objectives To synthesize hepatitis B virus e gene which was suitable for yeast protein expression and express hepatitis B virus e antigen (HBeAg) in Pichia pastoris.Methods Using synonymous codons preferred by yeast usage on protein expression to replace some native codons of wild-type HBV e gene,the synthetic e gene (syneAg-gene) was achieved by a recursive PCR (rPCR).The syneAg- gene was inserted into the yeast expression vector pPICZ?A.The recombinant plasmid was transformed into GS115 yeast by electroporation.The yeast transformant induced by methanol expressed the HBeAg.Results The restriction analysis and DNA sequencing confirmed that the syneAg-gene was inserted to yeast pPICZ?A in correct orientation.SDS-PAGE,immunoblot and ELISA indicated that the secreted form of HBeAg was expressed by the yeast transformant.The expression level of HBeAg in the strain containing syneAg-gene was 63 mg/L.The titer of the recombinant HBeAg in culture superuatant was 1 :81 920 and the maximum OD (optical density) value of absorbance was 2.8 by ELISA.No cross reactivity between Pichia pastoris-derived HBeAg and anti-HBc antibody was found.Conclusion The recombinant HBeAg with high degree of specificity and immunoreactivity is expressed efficiently in Pichia pastoris.

OBJECTIVETo give some theory support of Cistanche tubulosa cultivation by searching dry matter accumulation and echinacoside content of C. tubulosa.

METHODDry matter accumulation content of C. tubulosa culturing in Huabei plain was analysed in different growth season of C. tubulosa. Echinacoside content was determined by HPLC.

RESULTDry matter accumulation of C. tubulosa showed "S" variation. Dry matter accumulation increased fastest in September among growing seasons. Dry matter amount was 138.58 g after C. tubulosa grew a year. Dry matter amount decreased significantly along with inoculation time retarded. Echinacoside content was 30.59% when C. tubulosa grew in 5 months, decreased guadully after that, and 9.76% in annual.

CONCLUSIONVariation rule of dry matter accumulation and echinacoside content was found in C. tubulosa that grew one year in Huabei plain.

Biomass ; China ; Cistanche ; chemistry ; growth & development ; Glycosides ; metabolism ; Plants, Medicinal ; chemistry ; growth & development ; Seasons

Objective A resistin binding peptide (RBP) was selected by phage display in our previous work. Studies had shown that RBP could antagonize the role of resistin on the lipid metabolism and endocrine function of adipose tissue, but whether RBP affects the insulin secretion of pancreatic cells is still unknown. The aim of this study is to assess the effect of RBP on basal insulin secretion in RINm5F insulinoma cells. Methods The cell viability was measured by 3-[4,5-dimethyhhiazol-2-yl]-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay. The supernatants were assayed for insulin content by enzyme linked immunosorbent assay (ELISA). Reverse transcriptase-PCR assay and Western blotting were used to determine the expression of glucose transporter 2 (GLUT2) involved in insulin secretion. Cytosolic Ca2+, the trigger of insulin exocytosis, was analyzed with the fluorescent probe FURA-3/AM. Results RBP did no effect on the cell viability with a concentration of 10-8-10-12mol/L of 2 hours intervention. But it stimulated basal insulin secretion of RINm5F cells, accompanied by up-regulated increased expression of GLUT2 and elevated concentration of cytosolic Ca2+. Conclusion RBP could stimulate basal insulin secretion without affecting the cell viability.