Objective To clone and compare RNase HIIa and RNase HIIb of Chlamydia pneumoniae AR39(CpRNaseHIIa and CpRNaseHIIb). Methods Genes of CpRNase HIIs were amplified with the designed primers.Then,the recombinant plasmids were constructed,and CpRNase HIIs were expressed and purified with pET expression system.The 5′-32P-labeled RNA/DNA substrate and DNA with oligoribonucleotides substrates were prepared to identify characterization of CpRNase HIIs. Results Ribonuclease H activity of both CpRNase HIIa and CpRNase HIIb could cleave oligoribonucleotides strand,generating break with 3′-OH and 5′-phosphate ends.The results showed that there was different biochemical characterization between them. Conclusion CpRNase HIIa and CpRNase HIIb have different functions in C.pneumoniae.

Objective To investigate the effect and mechanism of adenovirus vector mediated SOCS 3 gene transfection in CD4+ Th cell differentiation and expression of inflammatory cytokines of mouse .Methods The CD4+ Th cells were isolated from spleen of C57bl/6 mouse and cultured .Ad‐SOCS3 were transfected into the CD4+ Th cells .PHA was used for culturing with the CD4+ Th cells .RT‐PCR were used to detect the mRNA expression ,and Western blot were used to detect the protein expression of cytokines .Results Compared with the control group ,the gene and protein expression of T‐bet ,IL‐2 ,IFN‐γ,STAT4 and IL‐12Rβ2 in the transfected group were significantly down‐regulated ,the gene and protein expression of SOCS3 ,GATA‐3 ,IL‐4 ,IL‐6 ,IL‐10 and STAT6 were significantly up‐regulated(P<0 .01) .Conclusion The results indicate that SOCS3 gene transfection can up‐regu‐late SOCS3 mRNA and protein expression in the CD4+ Th cells ,down‐regulate the JAK/STAT pathway ,inhibition of Th1 cell dif‐ferentiation ,and down regulation of inflammatory cytokine gene and protein expression ,and indirectly promote Th2 cell differentia‐tion ,and up the corresponding inflammatory cytokine gene and protein expression .

Objective To introduce our experience of performing posterior or anterior renal lip incision for partial nephrectomy in the treatment of renal hilum endophytic renal cell carcinoma.Methods From Jan.2010 to Jan.2014,five female patients with renal hilum tumors were treated in our institute.The median age was 54 (51-72) years.The median tumor diameter was 4.0 (2.8-4.8) cm.The median preoperative creatinine was 53.9 (52.6-75.4) μmol/L.One of them was solitary kidney with absolute indication; three cases had basic disease with relative indication; one was with selective indication.The patients were put in supine or lateral position.After general anesthesia,we preformed partial nephrectomy by cutting posterior renal lip in 3 cases and the anterior lip in 2 cases.We clamped the renal artery,opened the renal posterior or anterior lip,then dissected the tumor beside the pseudo-capsule.After removing the tumor,we used 3-0 absorption suture to control bleeding and repair the opened collecting system.Finally,we used 2-0 absorption suture to close the renal defect.Results The median operation time was 195 (155-215) min; the median renal warm ischemia time was 35 (15-70) min; the median postoperative hospital stay was 8 (7-9) d.There was no secondary bleeding and urine leakage happened.The pathological results showed that 3 cases with clear cell carcinoma,1 with papillary carcinoma and 1 with renal medullary interstitial cell tumor.All patients showed normal kidney shape.The median postoperative creatinine was 63.0 (59.4-75.4) μmol/L.After a median follow up of 24.2 mon,all patients survive without tumor recurrence.Conclusions The short-term result of posterior or anterior renal lip incision partial nephrectomy in treating endophytic renal hilum endophytic renal cell carcinoma is safe and feasible.

This paper focuses mainly on the study of optimal fermentation conditions of S-adenosyl-L-methionine.Effects of carbon sources,nitrogen sources,inorganic constituents,growth factors and adding time of L-methionine on the yield,the content and biomass of S-adenosyl-L-methionine are studied.And ingredients of the culture medium are also optimized by the method of uniform design.The final optimum culture medium contains: glucose 30 g,Yeast powder 11 g,(NH_4)_2SO_4 12 g,K_2HPO_4?3H_2O 5 g,KH_2PO_4 10 g,MnSO_4?H_2O 0.09 g,ZnSO_4?7H_2O 0.14 g,MgCl_20.5 g,CaCl_2 0.3 g,CuSO_4 0.005 g per liter. Using that optimum culture medium,the yield of S-adenosyl-L-methionine can reach 0.9 g/L in Erlenmeyer flask which is 30 % higher than before.Experiment on 5 L fermenter reveals that the accumulation of S-adenosyl-L-methionine can reach 2.66 g/L.Biomass is 23.4 g/L.

Objective To establish KIF4A 3'UTR esiRNA library and analyze the advantage of method in studying the function of KIF4A in SGC-7901 cells.Methods The GST-fusion protein of Escherichia coli endoribonuclease Ⅲ (GST-RNase Ⅲ) was used to digest the double strand RNA (dsRNA),which was transcribed in vitro from a 502bp template of KIF4A genome (T7-KIF4A 3'UTR).The KIF4A esiRNA library generated from the above method and chemically synthesized KIF4A siRNA were then used to transfect SGC-7901 cells at 10 nmol/L and 20 nmol/L.Real-time quantitative PCR and Western Blot were used to detect the mRNA and protein level of KIF4A,respectively.Results The KIF4A esiRNA library was effectively established from dsRNA digestion using GST-RNase Ⅲ.KIF4A expression was significantly reduced in SGC-7901 cells transfected with KIF4A esiRNA or siRNA.In addition,the inhibitory effect of KIF4A esiRNA was more effective than that of chemically synthesized siRNA.Conclusion KIF4A esiRNA library which was obtained using biological method can more effectively inhibited the expression of KIF4A more effectively in SGC-7901 cells than chemically synthesized siRNA.Therefore,esiRNA library can be used as a new and more effective method in studying the function of KIF4A in SGC-7901 cells.

Cells, Cultured ; Coal ; toxicity ; Dust ; Humans ; In Vitro Techniques ; Mutagenicity Tests ; Mutagens ; toxicity ; Mutation ; drug effects ; Salmonella typhimurium ; drug effects ; genetics ; Sister Chromatid Exchange ; drug effects

Adult ; Aged ; Aged, 80 and over ; Coal Mining ; Humans ; Lung Neoplasms ; complications ; microbiology ; Male ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; complications ; microbiology ; Staphylococcus aureus ; drug effects ; isolation & purification

Objective To explore the expression of nemo-like kinase (NLK) in primary hepatic carcino-ma (HCC) and its clinicopathological significance. Methods The expression of NLK was detected in 136 HCC samples by Immunohistochemistry. Results NLK expression was significantly up-regulated in HCC specimens compared to corresponding normal liver tissues. High expression of NLK was significantly associated with Ed-mondson-steiner grade, tumor size and number of tumor nodules (all P < 0.05). There was positive correlation between NLK and proliferation marker Ki-67 (P < 0.01). Kaplan-Meier analysis showed the high expression NLK group was significantly associated with poor overall survival and disease-free survival (all P < 0.001). Univariate analysis showed that the high expression NLK was associated with poor prognosis (P < 0.001). Multivariable Cox regression analysis suggested that the expressions of NLK and Ki-67 , Edmondson-steiner grade , metastasis , tu-mor size and number of tumor nodules were independent prognostic indicators for HCC. Conclusions NLK was markedly upregulated in HCC specimens, and it might be an independent prognostic marker for HCC. NLK might play an important role in tumorigenesis, progression and prognosis of HCC.

[ ABSTRACT] AIM:To investigate the effects of microRNA-378*( miR-378*) on the survival and apoptosis of human mesenchymal stem cells ( hMSCs ) .METHODS: The expression of miR-378* was determined by microRNA arrays and quantitative real-time PCR ( qRT-PCR) .H2 O2 was used to induce hMSCs apoptosis.By transfection of miR-378*mimic or inhibitor, we up-regulated or down-regulated miR-378* expression in hMSCs.The effect of miR-378*and connective tissue growth factor ( CTGF) on hMSC survival and apoptosis were detected by MTT, LDH, caspase-3/7 and TUNEL assays.RESULTS:The expression of miR-378*was up-regulated in the old hMSCs compared with the young hMSCs.H2 O2 increased the expression of miR-378*, decreased the expression of CTGF.Up-regulation of miR-378*re-sulted in increasing apoptosis and decreasing survival of hMSCs.Conversely, down-regulation of miR-378*resulted in de-creasing cell apoptosis and increasing survival.The regulation of miR-378*on hMSC apoptosis and survival was attenuated by inhibiting the expression of miR-378* and CTGF together.Direct repression of CTGF expression inhibited the hMSC survival and increased apoptosis.CONCLUSION:miR-378*enhances apoptosis of hMSCs by repressing the expression of CTGF.

Objective To evaluate the effectiveness of suberselective uterine arterial embolization for uterine fibroids.Methods Uterine arterial embolization with golyvimylalcohol(PVA) particles or Iodized oil and Gelfoam or Pingyangmycin lipiodol and Gelfoam was performed in 182 patients with uterine fibroids.Results Bilateral and unilateral superselective uterine arterial embolization were performed in 173 cases and 9 cases respectively. 6～28 months (mean 11 months) after the procedure, complete disappearance of tumor(16 cases), an average shinkage of 67% in tumor volume(152 cases) and a mean 42% reduction of uterine volume were obtained in 168 followed-up cases. The clinical symptoms were relieved significantly.The main side effets were hypogastic pain(135/182).Conclusion Superselection uterine arterial embolization is an effective and microinvasive method in treating uterine fibroids.