Objective To observe changes of cardiorespiratory function and blood gas analysis in different pneumoperitoneum pressure during age-specific pediatric laparoscopic surgery and to find a compatible pneumoperitoneum pressure for different age-specific children.Methods One hundred and fifty cases undergoing laparoscopic surgery were divided into three groups:A(1～3y),B(4～6y),C(7～12y).Each group was divided into two subgroups again by different pneumoperitoneum pressure:A1,A2,B1,B2,C1,C2.Monitoring the cardiorespiratory function and blood gas analysis in different time point before and after pneumoperitoneum.Results Compared with the baseline value,PETCO2,Paw,MAP,HR,PaCO2 significantly increased after peritoneal insufflation in each group.And the increase in A2,B2,C2 were more obvious than in A1,B1,C1 (P<0.05).The eligible pneumoperitoneum pressures for A,B,C were 9,11,13mm Hg respectively.Conclusion Using a compatible pneumoperitoneum pressure for different age-specific children can meet the requirement of operation and attenuate the interference with physiological function.

Objective: To construct the recombinant eukaryotic expression plasmid pHsp70-hsv-tk(pHT)for further target gene therapy research.Methods: The oligomers of primers were synthesized based on the reported sequences of hsv-tk and with the pHSV-106 plasmid as the template.Then the hsv-tk gene fragment was amplified and inserted into the pD3SX vector.E.coli DH5? was transformed with the recombinant plasmids and the bacterial colonies containing the recombinant plasmids were identified by the PCR cloning method,and the recombinant plasmids were extracted and purified.Results: The recombinant eukaryotic expression plasmid pHT was obtained.The target gene obtained by PCR amplification had the same molecular size as predicted.The recombined pD3SX plasmid contained correct recombinant hsv-tk sequences.Conclusion: The plasmid pHT was successfully constructed,which can be used for further target gene therapy research.

Objective To investigate surgical outcomes of treatment of pilon fractures of Ruedi -Allgower types Ⅱ and Ⅲ by raft screws fixation combined with bone grafting.Methods 21 patients with pilon fractures of Ruedi -Allgower types II and Ⅲ received surgery were selected.They were 15 men and 6 women,23 to 68 years of age(average 38.2 years).16 cases were closed and 5 cases open.12 cases were type II and 9 cases type III.Intervals between injury and operation ranged from 7 to 14 days,with 8.5 days on average.All of them were treated with raft screws fixation by plate type L combined with bone grafting.Results All cases were followed up for an average of 12 months(range from 8 to 30 months).Fractures healed from 16 to 24 weeks,with 18.2 weeks on average.According to the Burwell -Charnley radiological criteria,anatomical reduction was achieved in 17 cases,and fair reduction in 4.According to the Mazur clinical criteria,ankle functions were rated as excellent in 8 cases,good in 10 cases,fair in 2 cases and poor in 1 case.The rate of excellent and good was 85.7%.Conclusion Raft screws fixation combined with bone grafting are effective for the pilon fractures of Ruedi -Allgower types II and Ⅲ,preventing articular surface collapse and leading to anatomic reduction,and has stable fixation and early functional rehabilitation.

Objective To evaluate the clinical outcome of artificial cervical disc arthroplasty combined with anchoring cervical intervertebral fusion cage (ACIFC) in the treatment of multilevel cervical disc herniation. Methods From October 2011 to September 2013, 0.26 patients with multilevel cervical disc herniation underwent artificial cervical disc arthroplasty combined with ACIFC, there were 10 male and 16 female with age from 32 to 60 years (mean 46.8 years). Two segment were involved in 15 patients, three segment were involved in 11 patients. Neck disability index (NDI) and Japanese Orthopaedic Association (JOA) score were used to evaluate neurofunctional recovery preoperatively and postoperatively. The postoperative stabilization and the range of motion (ROM) of implanted disc and the fusion of cage were observed on dynamic radiograph. Results The average operation time and amount of bleeding were (95±15) min and (100±20) ml respectively. The average postoperative follow-up was 15 months(range 6-20 months). At the end of follow up,the scores of JOA was increased and the scores of NDI was decreased:(9.08±1.72) scores vs. (14.88±1.37) scores, (40.31±4.97) scores vs. (25.23±4.10) scores,there had significant differences (P<0.05). The change of ROM had no significant difference ( P>0.05). Solid fusion was achieved in all levels brought by cage.There was no cage subsidence or displacement. All artificial cervical disc achieved definite stabilization, no intraoperative and postoperative complication was noted. Conclusions Artificial cervical disc arthroplasty combined with ACIFC provides a feasible way for the treatment of multilevel cervical disc herniation. The definite stabilization and maintained ROM can reduce the fusion segments and avoid the increased stress of adjacent segment which can ensure a good preliminary clinical outcome.

Objective:To study the anti-proliferation effects of a heat shock protein 90(Hsp90) inhibitor,17-allylamino-17-demethoxygeldanamycin(17-AAG),on a human breast cancer cell line,SKBr3,and related mechanism.Methods:MTT assay was used to detect the growth inhibition of SKBr3 cells.Cell cycle and apoptosis were analyzed by flow cytometry.Alteration of human epidermal growth factor receptor 2(HER2) in SKBr3 cells being treated with 17-AAG were measured by immunohistochemistry.Results:17-AAG significantly inhibited growth of SKBr3 cells in vitro in a dose-dependent manner with an IC_(50) value at 3.09 ?g/ml.Under concentrations of 0,0.625,1.250,2.500,5.000 and 10.000(?g/ml,)the percentages of cell apoptosis were(1.03?0.08)%,(3.68?0.67)%,(7.06?1.12)%,(11.23?1.36)%,(20.32?1.98)%,and(31.65?2.96)%;the percentages of cells at G_(0)/G_(l) phase were 58.61%,54.34%,49.55%,43.73%,35.52%,and 27.46%;the percentages of cells at S phase were 29.57%,25.21%,19.65%,22.98%,19.71%,and 15.46%;the percentages of cells at G_(2) /M phase were 11.82%,20.45%,30.18%,33.29%,44.77%,and 57.08%,respectively.The level of HER2 expression in SKBr3 cells being treated with 17-AAG,compared to that in control cells,was reduced significantly.Conclusion:17-AAG can inhibit the growth of human breast cancer cell and enhance its apoptosis.It may be a promising anti-tumor drug.

Objective To investigate the effects of isotope labeled folate targeting albumin nanoparticles (188Re-fo-late-CDDP/HAS MNP) on human SKOV3 ovarian cancer cells in vivo. Methods The human SKOV3 ovarian cancer model was established in mice. Sixty-four tumor-bearing mice were randomly divided into eight groups:(A) negative control group, (B) chemotherapy group, (C) radiotherapy alone group, (D) hyperthermia alone group, (E) chemotherapy combined with radio-therapy group, (F) chemotherapy combined with hyperthermia therapy group, (G) radiotherapy combined with hyperthermia therapy group and (H) hyperthermia, chemotherapy and radiotherapy combined treatment group. After treatment, the cell pro-liferation and tumor growth were observed. The inhibitory rate of tumor mass was measured. The histopathological changes of tumor were observed in all groups. Results The quality of tumor was significantly lower in treatment groups than that of control group (P＜0.05). There was the lowest quality of tumor in hyperthermia, chemotherapy and radiotherapy combined treatment group than that of other treatment groups (P＜0.05). Conclusion The combination of magnetic induction hyper-thermia, chemotherapy, targeted radionuclide of radiation exposure can effectively inhibit the growth of ovarian cancer, which has the potential application for ovarian cancer treatment.

Using Eu3+as a tracer,a sandwich?type assay was established. HE4 in serum specimens from 225 patients were detected by TRFIA. Serum levels of HE4 in pelvic tumors were quantitatively analyzed. From receiver operating characteristic (ROC) curves,the reference values was calculated for endometrial cancer,cervical cancer and ovarian cancer,respectively. Results The working range of serum HE4 for TRFIA was 10~10000 pmol/L with a sensitivity of 7.5 pmol/L. The method offered less interaction with CA125 and CA199,and also provided a better correlation with ECLA,while the CV of intra?assay was below 10%. HE4 levels were significantly higher in endometrial cancer,cervical cancer and ovarian cancer groups than in healthy volunteer group,while did not differ significantly between uterine fibroid group and healthy women group. As aging,the risk for developing pelvic tumors increased. The area of ROC for malignant tumors was all above 0.5 and the accurate interpretation of HE4 was 60 pmol/L. Conclusion A stable,precise HE4 TRFIA is well established,which is helpful in diagnosis of gynecologic cancers.

Objective To compare the efficacy and safety of Iamivudine-interferon sequential therapy and lamivudine monotherapy in HBeAg-positive chronic hepatitis B (CHB) patients.MethodsA total of 172 patients with HBeAg-positive CHB were randomized to sequential group (n=83) or lamivudine group (n=89).Sequential group were administrated with lamivudine 100 mg/d and 5 million units interferon alpha 2b subcutaneous injection every other day for 24 weeks were added since week 25 of treatment.Lamivudine group were administrated with lamivudine 100 mg/d for 48 weeks.All subjects were followed up for 24 weeks after drug withdrawal.Measurement data with homogeneity of variance were analyzed by using t test and data with heterogeneity of variance were analyzed by using rank sum test.The comparison of rates was done by chi square test or Fisher exact test.ResultsThe baseline hepatitis B virus (HBV) DNA levels of patients in sequential group and lamivudine group were (7.8±1.0) and (7.9±1.1) lg copy/mL,respectively (P＞0.05),and the baseline alanine aminotransferase (ALT) levels were (210.5 ± 150.1 ) and (211.9 ± 160.9) U/L,respectively (P＞0.05).At the end of treatment,higher ALT levels [(78.4±146.1) vs (36.1±32.4) U/L,P＜0.05)] and HBV DNA levels [(4.5±1.5) vs (3.8±1.3) lg copy/mL,P＜0.05)] levels,lower response rates (65.8％ vs 83.5％,P＜0.05),and similar HBeAg loss rates (31.6％ vs 22.2％,P＞0.05) and HBeAg seroconversion rates (27.6％ vs 16.0％,P＞0.05) were found in sequential group compared with lamivudine group.At the end of follow-up,higher ALT levels [(126.0±143.1) vs (82.7±83.0) U/L,P＜0.05)],similar HBV DNA levels [(5.3±1.5) vs (5.0±1.5) lg copy/mL,P＞0.05)],similar HBeAg loss rates (25.0％ vs 32.3％,P＞0.05) and HBeAg seroconversion rates (25.0 ％ vs 26.2 ％,P＞0.05) were found in sequential group compared with lamivudine group.YMDD motif mutation rate in sequential group was lower than lamivudine group at week 48 of treatment (10.5％ vs 26.9％,P＜0.05).ConclusionsLamivudine-interferon sequential therapy and lamivudine monotherapy are both effective in HBeAg-positive CHB patients,while HBV mutations are reduced in patients with sequential therapy.

Objective To prepare 188 Re-c ( CGRRAGGSC) by different labeling methods, and compare the stability and targeting ability for triple-negative breast cancer (TNBC). Methods 188 Re la-beled c( CGRRAGGSC) was prepared by direct method ( pre-tinning method) and indirect method (the hydrazinonicotinamide (HYNIC) conjugated c(CGRRAGGSC) peptide) respectively. Quality control and sta-bilities of radiolabeled probes were measured. Nude mice bearing TNBC cells (MDA-MB-468) were estab-lished and used for detecting the distribution in vivo. Tumor/ non-tumor (T/ NT) ratios of radiolabeled probes prepared by direct method and indirect method were compared to certify the targeting ability and biological activi-ty. Two-sample t test was used to analyze the data. Results The labeling rate of 188Re-c(CGRRAGGSC) by di-rect method was >87％, but the serum stability was poor (the degradation rate was about 40％ at 2 h), and the T/ NT ratio was 2.82±0.23 (n=3) at 2 h. On the other hand, the labeling rate of 188Re-HYNIC-c(CGRRAGGSC) by indirect method was 61％, while the serum stability was about 10％ at 2 h, and T/ NT ratio was about 6. 27±0.51 (n= 3) at 2 h, which was significantly higher (t = 2.13, P<0.05). Conclusions The direct method may have higher labeling rate than the indirect method, but the radiolabeled compound by the direct method has poor stability and tumor targeting ability. 188 Re-HYNIC-c(CGRRAGGSC) prepared by indirect method may have remarkable advantages on stability and biological activity.