Effects of RNA Interfering of MBP-1 on Proliferation of Saos-2 Cell Line / 中国医科大学学报

Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.

Construction of shSet7/9 vector and its function in HepG2 / 西安交通大学学报(医学版)

Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.

Clinical study on the treatment of ectopic pregnancy by methotrexate and the particle of gelatin sponge with uterine artery embolization / 中国医师杂志

Objective To investigate the clinical application value of methotrexate and the particle of gelatin sponge treat ec -topic Pregnancy .Methods A total of 48 cases of ectopic pregnancy was analyzed retrospectively .Methotrexate and gelatin sponge particles were injected into the uterine artery embolization for the method of intervention , and its clinical application value was evalua-ted.Results All 48 patients were embolized uterine artery successfully .It treated successfully ectopic pregnancy 44 cases (92%), including 38 cases of tubal pregnancy and other parts of the 6 cases.All patients were detected in β-human chorionic gonadotropin (β-HCG) until normal about 3 weeks.No serious postoperative complications were found , only 30 patients with abdominal pain, 16 cases of patients with nausea and vomiting , 9 patients with low-grade fever were found .After four months , 29 patients were recanalizated successfully .Conclusions Treatment of ectopic pregnancy can kill embryos efficiently , stanch the bleeding rapidly , and have small operation wound .It is safe and reliable method and is worth populating .

Evaluation of the model for end-stage liver disease and Child-Turcotte-Pugh score on the surgical prognosis in patients with liver cirrhosis / 中国医师进修杂志

Objective To investigate the value of the model for end-stage liver disease (MELD) and Child-Turcotte-Pugh (CTP) score in predicting the prognosis of patients with liver cirrhosis.Methods Seventy patients with liver cirrhosis were selected.The MELD and CTP score before surgery was calculated and was analyzed the correlation between the two models was analyzed.The prognosis ability by the area under the receiver operating characteristic (ROC) curve was evaluated.Results Twenty three cases (32.9％,23/70) appeared post operative serious complication.The scores of MELD and CTP in complication group (23 cases) was (19.58 ±5.90),(8.84 ± 1.87) scores,the scores of MELD and CTP in without complication group (47 cases) was ( 12.27 ± 2.94),(6.10 ± 1.12) scores,there were significant differences between two groups (P ＜ 0.01 ).According to the MELD score,70 patients was divided into ＜ 14 scores group(30 cases),14 - 23 scores group(28 cases),＞ 23 scores group( 12 cases),the rate of complication was 10.0％(3/30),35.7％( 10/28 ) and 83.3％(10/12),there were significant differences among three groups(P＜ 0.05).According to the CTP score,70 patients were divided into A grade(29 cases),B grade (25 cases) and C grade( 16 cases),the rate of complication was 10.3％ ( 3/29 ),36.0％ (9/25) and 68.8％ ( 11/16 ),there were significant differences among three groups (P ＜ 0.05 ).The results of Pearson correlation analysis showed that the MELD score and CTP score had significant correlation (r =0.874,P ＜ 0.01 ).The area under the ROC curve of the MELD score and CTP score in prognosis the perioperative complication was 0.877 (95％ CI:0.84 - 0.95 ) and 0.852 (95％ CI:0.83 - 0.94),there was no significant difference ( U =0.157,P ＞ 0.05 ).Conclusion Both MELD and CTP score can accurately predict the short term prognosis of patients with liver cirrhosis.

Cloning of MSI-78 Gene in Escherichia coli DH5? and Identification of Positive Recombinant / 中华医院感染学杂志

OBJECTIVE To clone the MSI-78 gene for the purpose of providing evidence for further studies in prokaryotic expression and activities of antimicrobial peptides. METHODS According to the amino acid sequences of MSI-78,the MSI-78 gene was designed favorable for the Escherichia coli codons. After EcoRⅠand PstⅠ disgestion,cohesive ends were added to both ends respectively and the MSI-78 gene was synthesized by chemical methods. Then,the MSI-78 gene was ligated with pUC-18,transformed into the E. coli DH5?. Through filtration of ? complementary screening,the positive recombinant was finally identified by enzyme digestion of ECORⅠand ECORⅠ/PstⅠ and by PCR. RESULTS The MSI-78 gene was ligated with pUC-18 and transformed into the E. coli DH5?. As a result,MSI-78 gene was cloned in E. coli DH5? successfully. CONCLUSIONS The cloning of the MSI-78 gene provides evidence for further studies of its prokaryotic expression and activities of antimicrobial peptides.