Humans ; Hypertension, Pulmonary ; complications ; physiopathology ; Ventricular Dysfunction, Right ; etiology

Objective: To chine and express the recombinant human endothelial monocyte-activating polypeptide-Ⅱ(EMAP-Ⅱ)and identify its anti-tumor biological activities.Methods: EMAP-Ⅱ_(147-312)was expressed by the expression vector pMAL-p2x and E.coli BL-21 and the product was purified.The production of tissue factor(TF)in human umbili- cal vein endothelial cell ECV-304 mediated by the recombinant EMAP-Ⅱwas determined by chemiluminescence sub- strate.The promoting effect of recombinant EMAP-Ⅱon TNF?-induced ECV-304 cell.Apoptosis was determined by flow cytometry.Its inhibitory effect on human pancreaic cancer cell SW1990 proliferation was determined by MTT method. Results:DNA sequencing verified that EMAP-Ⅱwas correctly cloned.The molecular mass of the protein identified by SDS-PAGE was consistent with the theoretic value.The productivity of recombinant EMAP-Ⅱwas 500?g per 1 g bacteria (wet mass).The purified product induced expression of tissue factor(TF)in ECV-304 cells;it also enhanced the sensi- tivity of ECV-304 cells to the apoptotic effect of TNF?([16.6?2.5]% vs[25.6?2.3]%,P

Objective To investigate the numbers and the expression of MHC-Ⅰpositive cells in hu- man ovarian epithelial carcinoma tissues and provide the experimental data in the futher biological therapy of ovarian carcinoma.Methods Thirty one samples of ovarian epithelial carcinoma were analysed with flow cy- tometry for MHC classⅠexpression.Results The number of MHC classⅠcells(25.22?21.23,4.37?3.63)was fewer in ovarian epithelial carcinoma than that in normal ovarian tissues(43.56?18.47,7.43?5.87),and was related with pathologic grades(P

Objective To study the effect of the extract of Chrysanthemum Indicum(CI)on the expression of tumor necro-sis factor-α (TNF-α) and neutrophil function in chronic bronchitis (CB)rats and to explore the therapeutic mechanism for chronic bronchitis. Methods The extract of CI was prepared. Wistar rats were randomly divided into blank control group, model group, Chuanxiling group, and low-, middle-and high-dose CI extract groups. Rat model of CB was established by intratracheal injection with low-dose lipopolysaccharide. After drug intervention, the expression of TNF-α in rat serum and bronchoalveolar lavage fluid was detected by ELISA method. Phagocytic function of the neu-trophil and respiratory burst of the rats were measured by flow cytometry (FCM). Results Compared with the blank con-trol group, the phagocytic function of the neutrophil, respiratory burst of the rats, and the expression of TNF-α in rat serum and bronchoalveolar lavage fluid of the model group were increased significantly. CI extract significantly decreased the abnormal rising of the above indexes. Conclusion Down-Regulation of TNF-α expression, and decrease of the neutrophil phagocytic function and rat respiratory burst may be one of the therapeutic mechanisms of Chrysanthemum In-dicum extract for the treatment of CB.

The tissue-specific and MeJA-induced transcriptional levels of BcUGT3 and BcUGT6 in Bupleurum chinense were analyzed in the present study. The transcriptional levels of BcUGT3 in root, leaf, flower and fruit were similar and they all were higher than those in stem. The transcriptional level of BcUGT6 was the highest in leaf and the lowest in flower among in all tested tissues. With non-treated adventitious roots as control, BcUGT6's transcriptional levels were elevated to nearly 2 folds for 2 h, 8 h, 24 h, 2 d and 4 d in MeJA-treated adventitious roots of B. chinense. It showed that the transcriptional level of BcUGT6 was slightly affected by MeJA. While, BcUGT3's transcriptional levels were gradually elevated, and till 4 d after MeJA treatment, the expression level was about 7 folds than that of non-treated control. Using pET-28a (+), the expressions of two genes was investigated. Induced by IPTG, the target proteins were expressed in E. coli and then purified. All the results obtained in the present study will be helpful for follow-up bio-function analysis of BcUGT3 and BcUGT6.
Acetates ; pharmacology ; Bupleurum ; cytology ; enzymology ; genetics ; Cell Membrane ; metabolism ; Cyclopentanes ; pharmacology ; Escherichia coli ; genetics ; Gene Expression ; Gene Expression Regulation, Plant ; drug effects ; Hexosyltransferases ; chemistry ; genetics ; isolation & purification ; metabolism ; Intracellular Space ; metabolism ; Oxylipins ; pharmacology ; Protein Sorting Signals ; Protein Structure, Secondary ; Protein Transport ; Sequence Analysis ; Transcription, Genetic ; drug effects

The high molecular weight genomic DNA of yeast was extracted using three methods.Products were separated on agarose gel electrophoresis,quantified by spectrophotometer ND-1000 and restricted by EcoRⅠand MesⅠ.The result was shown that the genomic DNA extracted by modified benzyl chloride method was the best.The products of wild isolates supported it,too.This method was suitable for restriction of genomic DNA from yeast.

Objective To discuss the correlation between cervical intraepithelial neoplasia(CIN) and uterine cervix cancer and the combination of TCT and colposcope for the investigation of cervical lesion.Methods 5545 patients were preliminarily screened by cervical fluid basement cell folium smear,in which 307 patients are masculine and 219 are negative,the 526 patients are suspected cervical lesions in clinical symptoms and they further underwent colposcopy and biopsy,the results were analyzed by pathohistology which is golden standard.Results CIN coherent dangerous factors analyzed by single factor Logistic regression analysis are:cervical HPV pollution,age of sexual activity,sexual partners numbers,protection of sexual life, STD history,age and frequency of miscarriage ( P < 0.05 ).The coincidence between TCT,colposeopy and pathologic diagnosis are 49.4%, 83.8% ;the omission diagnostic rate of LSIL,HSIL and CC from TCT are 63.63% ,74.44% ,100% ;the omission diagnostic rate of LSIL,HSIL and CC from colposcopy are 33.37%, 12.22% ,0;the sensitivity of TCT,colposcopy and TCT combine colposcopy for CIN are 61.2% ,84.6% ,94.5% and specificity are 85.4% ,88.2.% ,90.8%.Conclusions (1)The generation of CIN and cervical cancer are correlated with cervical HPV pollution,sexual behavior and various kinds of lower genital tract infection.(2) Fluid basement cell folium smear can promote the property of diagnosis,meet the demands of early stage uterus neck cancer and precancer lesion investigation,are suitable to be routine cheek methods in primary hospital.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

Objective:To express Tumstatin_(183-230)-TRAIL fusion protein and to observe its biological functions.Methods: SOE-ing PCR was employed to amplify the recombinant sequence of Tumstatin_(183-230)and TNF-related apoptosis-inducing ligand (TRAIL_(114-281)).An expression vector pMAL-Tu-T was constructed by inserting Tu-T sequence into pMAL-c_2;the vector was used to transfect E.coli BL21(DE3)and expression of MBP-Tu-T fusion protein was induced by IPTG.Amylose Resin columns were employed to purify the fusion protein.The biological functions of MBP-Tu-T protein was examined by inhibitory test of endothelial cell proliferation,standard tumor cell cytotoxic assay,in vitro tube formation inhibition,and electron microscopic observation(apoptosis).Results:The expression rate of MBP-Tu-T fusion protein in E.coli was about 20%. Purified recombinant protein obviously inhibited endothelial cell proliferation(IC_(50)12.5?g/ml),induced apoptosis of pancreatic cancer cells,and inhibited tube formation.Conclusion:Constructed MBP-Tu-T fusion protein is bifunctional,which lays a solid foundation for further investigation of antitumor effect of Tumstatin_(183-230)-TRAIL in vivo.

AIM:To investigate the effect of phosphatidylinostiol 3-kinase ( PI3K) inhibitor GDC-0032 on cell viability, cell cycle and DNA damage in human glioma U 251 cells.METHODS:The cell viability was analyzed by MTT assay.The cell cycle distribution of U251 cells was examined by flow cytometry .The protein expression was examined by Western blot.The expression and localization of γ-H2AX were determined by laser-scanning confocal microscopy .RE-SULTS:GDC-0032 inhibited the cell viability in a dose-dependent manner .U251 cells showed G 1 phase arrest accompa-nied with upregulation of p 27 protein after exposure to GDC-0032, while the expression of cell division cycle protein 2 (Cdc2) was inhibited.GDC-0032 treatment increased the formation of γ-H2AX foci and histone H2AX phosphorylation in the U251 cells.In addition, GDC-0032 upregulated the phosphorylation levels of mitogen-activated protein kinases , inclu-ding extracellular signal-regulated kinase ( ERK) and c-Jun N-terminal kinase ( JNK) , in the glioma cells , while the ex-pression of survivin was inhibited .CONCLUSION:GDC-0032 inhibits U251 cell growth by inhibition of cell viability and cell cycle progression.GDC-0032 also induces DNA damage of U251 cells.The anticancer activity of GDC-0032 is associ-ated with increasing the activity of ERK and JNK and downregulating the expression of survivin .