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.

This experiment was aimed to create A20 gene site-specific zinc finger DNA-binding protein. The sequence of A20 gene promoter was analyzed with bioinformatics means and submitted to ZF Tools Server at TSRI. Using the database of the web site, we determined the A20 gene valid target sites and designed the amino acid sequence of zinc finger protein predicted to be bound to the target site. And then, the structure of the protein sequence was analyzed and homology was modeled with various bioinformatics means. Based on the characteristic of this protein, the prokaryotic expression vector pTYB11-ZFP was constructed and expressed. Thus, the artificial zinc finger protein that recognized A20 specific sequence was designed, and expressed in Escherichia coli. The results indicate that it is feasible to design engineered artificial Zinc finger proteins by means of bioinformatics.
Amino Acid Sequence ; Base Sequence ; DNA-Binding Proteins ; chemistry ; genetics ; Humans ; Molecular Sequence Data ; Protein Binding ; Protein Engineering ; methods ; Transcription Factors ; chemistry ; genetics ; Zinc Fingers ; genetics

Objective: To study the association between the AKAP12 promoter methylation and recurrence of hepatocellular carcinoma. Methods: A total of 142 primary liver cancer patients underwent surgery in department of Hepatobiliary surgery in Peking University Cancer Hospital from 2003 to 2009 were selected as subjects in the survey; with the inclusion criteria as hepatocellular carcinoma, no cancer cells were observed in the surgical margin(SM) samples. All patients had neither lymph nor distant metastasis at the time of surgery, and receiving complete follow-up data for at least 3 years. By the end of May 2014, a total of 75 patients had relapsed of whom 71 died and there were no lost. All samples were acquired from the frozen surgical tissues. Genomic DNA was extracted using phenol/chloroform method and performed bisulfite modification following with polymerase chain reaction (PCR). AKAP12 methylation in hepatoma and the corresponding SM samples from 142 patients was determined by denature high-performance liquid chromatography (DHPLC) and bisulfite clone sequencing. Kaplan-Meier and Cox proportion hazard regression model were used to identify the factors related to the survival time. Results: In 142 cases, 125 patients (88.0%) were male and 17 (12.0%) cases were female. The median age was 52.5 years, ranging from 34 years to 76 years. AKAP12 methylation-positive rate was significantly higher in hepatomas than SMs (54.9% vs. 10.2%, P<0.001). Patients with AKAP12 methylation-positive had less risk of the recurrence (HR=0.62, 95%CI: 0.39-0.99); with tumor diameter more than 5 cm (HR=1.53, 95%CI: 1.00-2.50),portal vein invasion(HR=4.53, 95% CI:2.69-7.64) increased the recurrence risk. Moreover, portal vein invasion had a higher risk of death (HR=2.98, 95% CI: 1.73-4.98). Conclusion There was significant association between AKAP12 DNA methylation and low risk of recurrence and long progression-free survival of hepatocellular carcinoma patients.