Objective To investigate the protective mechanism of trace element-selenium(Se) on the myocardium by against free radical superoxidation during myocardial ischemia/reperfusion(I/R) period in patients undergoing pulmonary bypass.Methods 46 patients with ASD or VSD were divided into control(23 patients) and Se administration(23 patients) groups.Before heart surgery,patients in the Se group orally received 400?g Se supplementation per day,for 7 days.The blood and myocardium samples were collected before clamping and after release respectively during pulmonary bypass.Biochemistry,electron spin resonance spectrometer(ESR),atomic absorption,RT-PCR and cDNA sequencing,were used to measure Se concentrations,MDA level,glutathione peroxidase(GPX) activity,GPX mRNA expression and GPX cDNA sequence in blood and myocardium.Results In Se group,before clamping,Se contents in blood cells and plasma, were not changed,but at reperfusion 30 minute,myocardial Se concentrations was evidently increased by about 1.1-fold,contrary to reduction 41% in blood cells.In Se administration group,at myocardial reperfusion 10 minute,free radical levels of the coronary sinus blood were significantly decreased 28%.No significantly changes in myocardial GPX activity or MDA level were found till reperfusion 30 minute.However,Se group had a higher myocardial GPX mRNA expression than that in controls in both before ischemia and after reperfusion.Conclusions Se supplementation can enhance Se content,expression of GPX-mRNA in myocardial tissue,and protect myocardium from radical lipooxidation damage.

Objective To investigate the relevant factors of neonatal ventilator-associated pneumonia(VAP),and to provide a theoretical basis of prevention and treatment.Methods Retrospective analyed the clinical data of 145 critically ill neonates,who were treated with mechanical ventilator from Jan 2006 to Dec 2009 in the Third People′s Hospital of Wenzhou City,NICU.According to whether the neonates were occurred VAP,they were divided into two groups:VAP group(52 cases) and without VAP group(93 cases).Results Fifty-two out of the 145 neonates developed VAP.The incidence of VAP was 35.86%,the main relevent factors were the gestational ages,birth weights,the duration of mechanical ventilation and the times of intubation.There was significant difference between the two groups (P<0.05).The main pathogens were opportunistic bacteria,and mostly were G-bacilli.Conclusion The incidence of VAP has a close conclusion with the gestational ages,the birth weights,the duration of mechanical ventilation and the times of intubation.Regulate the use of breathing machine,strengthen aseptic operation,and select effective antibiotic can control the occurrence and development of VAP.

Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.

Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.

Objective: To investigate the effects of antimicrobial peptide LL-37 on the tumor growth and apoptosis of the mice with colon cancer, and to elucidate the possible molecular mechanism of its anti-tumor effect. Methods: The LL-37 over-expression colon cancer HT-29 cells were constructed, and the expression levels of LL-37 mRNA and protein in the HT-29 cells were detected by qRT-PCR and Western blotting methods. A total of 30 BALB/c mice were randomly divided into control group (given the uninfected HT-29 cells)' empty vector group (given the HT-29 cells infected with empty plasmid), LL-37 over-expression group (given the HT-29 cells infected with LL-37 over-expression vector), AMPK inhibitor group [given the HT-29 cells infected with empty vector, and then injected with 2 mg • kg Dorsomorphin (Dor) in the tail vein^ and combination group (given the HT-29 cells infected with LL-37 over-expression vector, and then injected with 2 mg • kg Dor in the tail vein), and there were 6 mice in each group. The colon cancer HT-29 cells were used to replicate the colon cancer models of the mice. The tumor volumes of the mice in various groups were detected and the growth curve was drawn. After infected for 24 d, the weights of the tumor of the mice in various groups were measured and the inhibitory rates of the tumor were calculated. TUNEL staining was used to detect the apoptosis of tumor cells in various groups. Western blotting method was used to detect the expression levels of autophagy-related proteins Beclinl, ATG5, LC3 1, LC311 and AMPK/mTOR pathway-related proteins AMPK, p-AMPK, mTOR and p-mTOR in tumor tissue of the mice in various groups. Results: Compared with control group and empty vector group, the expression levels of LL-37 mRNA and protein in the HT-29 cells of the mice in over-expression group were significantly increased ( P< 0.01), the weight and volume of the tumor were significantly decreased ( P<0. 05)∗ the inhibitory rate of tumor was increased (P<0. 05)∗ the number of apoptotic cells was increased' the expression levels of autophage-related proteins Beclinl and ATG5 in tumor tissue and the ratios of LC3 II/LC3 1 and p-AMPK/AMPK were increased ( P<"0. 05), while the ratio of p-mTOR/mTOR was significantly decreased ( P<0. 05). Compared with empty vector group, the weight and volume of the tumor of the mice in AMPK inhibitor group were increased ( P<0. 05), the inhibitory rate of the tumor was decreased (P

Objective To explore the clinical efficacy of trastuzumab combined with vinorelbine therapy in the treatment of advanced breast cancer with human epithelial growth factor receptor-2 (HER-2) positive.Methods From April 2010 to April 2013,91 advanced breast cancer patients who failed in the paclitaxel therapy received trastuzumab plus vinorelbine (45 cases,vinorelbine group) or capecitabine (46 cases,capecitabine group).The treatment efficacy,toxic effects and 3-year survival rate in two groups were compared.Results The clinical benefit rate in two groups had no significant difference (P ＞ 0.05),but objective response rates in vinorelbine group was higher than that in capecitabine group:44.44％(20/45) vs.23.91％(11/46),and there was significant difference (P =0.039).The toxic effects in two groups had no significant difference (P ＞ 0.05).The 3-year survival rate in two groups had no significant difference (P =0.252).Conclusions In the treatment of HER-2 positive advanced breast cancer,trastuzumab plus vinorelbine or capecitabine shows no significant differences in adverse reaction or in 3-year survival rate.However,trastuzumab plus vinorelbine shows better objective response rate compared with trastuzumab plus capecitabine.

BACKGROUND:Recently, periprosthetic femoral fractures have been a common complication after total hip replacement. Vancouver type B1 periprosthetic femoral fractures commonly received internal fixation due to stable femoral prosthesis and less bone defects.
OBJECTIVE:To evaluate the effect of the application of Synthes cable system in the treatment of Vancouver type B1 periprosthetic femoral fractures after total hip replacement.
METHODS:From May 2009 to October 2012, 18 patients with Vancouver type B1 periprosthetic femoral fractures were treated with Synthes cable system in the Department of Orthopedics, Nantong First People’s Hospital. There were 8 males and 10 females with an average age of (62.67±8.67) years ranging 45 to 80 years. They received a further consultation at 1, 3 and 6 months after treatment. The function of hip joint was evaluated by Harrris evaluation standard:ful marks, 100 points, ≥ 90 points, excellent;80-89 points, better;70-79 points, good;<70 points, poor.
RESULTS AND CONCLUSION:Al 18 patients were fol owed up for 6 to 43 months. Mean healing period was (19.06±4.04) weeks. Harris score was (22.3±3.6) points before treatment in 18 cases. Harris score was higher significantly at 1, 3 and 6 months after treatment compared with that before treatment (P<0.01), and the recovery of hip function was good. Results indicated that Synthes cable system could help to restore the normal anatomic structure of femur in the greatest extent, showing simple operation, little injury, high stability and high safety. It is an ideal method for treating Vancouver type B1 periprosthetic femoral fractures.

Objective Clinical effect analysis of low dose dopamine(LDD)and dobutamine (DOB)subsidiary in the treatment of 86 cases of Children serious pneumonia.Methods From July 2011 to July 2013,86 CSP patients who were diagnosed with CSP in People's Hospital of Wenzhou were selected.Patientas were randomly divided into observation group(n=43)and control group(n=43)according to the random number method.Both groups were treated with routine treatment,then observation group on the basis with LDD and DOB subsidiary,then compared efficacy of two groups at 1 week after treatment,as well as the time of symptoms and signs which had improvements,and changes of levels of Interleukin-6(IL-6),Interleukin-8(IL-8), C-reactive protein(CRP)and Tumor necrosis factor-α(TNF-α)before and after treatment. Results The efficacy in observation group was significantly better than that of control group(P<0.05 ).And time of symptoms and signs which had improvement in the observation group was significantly less than control group(P<0.05).After treatment,IL-6,IL-8,CRP and TNF-αlevels were lower than before,but the observation group had a greater degree of reducing(P<0.05).Conclusion Routine treatment combined with LDD and DOB for CSP patients,can significantly improve the therapeutic effect,and relieve the clinical symptoms.

BACKGROUND:Serious patel ar comminuted fractures can be treated by a number of methods such as cerclage with tension band steel wire or steel wire, or fixation with the nickel titanium patel a concentrator and absorption lines, or cerclage with 10# double-loop wire plus fixation with split-type patel a claws or with the memory al oy patel a concentrator. But it is difficult to achieve strongly fixed effect on patel a fractures by these methods.
OBJECTIVE:To investigate the clinical effects of suture with polyester braided wire and cerclage with belt lock cable on serious comminuted patel ar fractures.
METHODS:A retrospective analysis was performed on the clinical data of 57 patients with serious comminuted patel ar fractures who received suture with polyester braided wire and cerclage with belt lock cable from January 2007 to October 2012. According to AO classification of fractures, there were nine cases of type B3, 22 cases of type C2 and 26 cases of type C3. After satisfactory reduction of comminuted patel ar fractures during the operation, firstly we sewed up both sides of the expansion region of quadriceps muscle and stitched on periosteum and prepatel ar tissue with thick polyester braided wire fol owed by patel ar cerclage with belt lock cable. According to the stability of fracture blocks, tension band fixation with polyester braided wire was partial y added. After the operation, the clinical effects were assessed based on the Bostman score system.
RESULTS AND CONCLUSION:Total y 46 out of 57 cases were fol owed up with an average of 8 months (4-18 months). Al patients obtained bone union within 3-5 months. Wounds healed in the first stage without any infections, and fractures in al patients healed without complications such as implant loosening and skin irritation. According to the Bostman score system, 33 cases were scored excellent, 10 good and three fair. The excellent and good rate was 93.5%. It suggests that treatment of serious patel ar comminuted fracture by the combination of polyester braided wire and belt lock cable leads to closely contact fracture sites and reliably fix bone fragments. It can facilitate early postoperative range of motion exercises.

The amino acid sequence (1-301aa) coding the human PTP1B catalytic domain (PTP1Bc) was obtained from the GenBank. The PTP1Bc gene was constructed by overlapping PCR, then was inserted into vector pET-22b(+) and expressed efficiently in E. coli BL21(DE3) under optimum condition after IPTG induction. The proteins were expressed mainly as inclusion bodies with the yield of more than 30% of total bacterial proteins. The expressed products were purified through Ni(2+)-affinity chromatographic column. After purification, the purity of the proteins was more than 95%. Western blotting analysis suggested that the purified proteins could specially combine with anti-PTP1B antibody. Enzyme activity assay showed that the protein has phosphatase activities.
Catalysis ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Inclusion Bodies ; metabolism ; Polymerase Chain Reaction ; methods ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism