Utilizing higher expressing vector and a synthetic oligonucleotide linker,a new expressing clone, pBMhIL 4, has been constructed in our laboratory. The intact human rIL-4 protein molecule was expressed under the control of pL promoter in E. coli. The gene encoding protein was expressed in the plasma of E. coli in the form of inclusion bodies, the expressed IL-4 constituting about 5-10% of the total cellular proteins by SDS-PAGE analysis. The molecular weight of human rIL-4 was about 15KD. After extraction and renaturation, the yieldedsoluble human rIL-4 exhibited biological activity. 1?10~6 units of human IL-4 were produced from 1 liter of bacteria extracts assassed by the assay of its TCGF activity. Using 3.5M Guanidine hydrochloride and 0.25% Triton X100 to lyze inclusion bodies resulted in higher yield and higher biological activity.

Utilizing Polmerase Chain Reaction (PCR) and recombination DNA techniques with synthetic oligonucleotide primers,a high level expressing clone for human IL- 6, pBMhIL6, was constructed successfully in our laboratory. The intact human rIL-6 protein molecular was expressed under the control of p_Rp_L promoters, synthetic SD sequence and the terminal codon TAA which replaced TAG of original human IL-6 in E. coli. the expressed human rIL-6 protein, molecular wight 21 KDa, with specific binding to Anti-IL-6 McAb, constituted about 28% of the total cellular proteins assessed by SDS-PAGE, densitometry analysis and Western Blot assay. The results of study on kinetics of inducing human rIL-6 expression in the defferent E. coli strains and influence of bacteria growth states ingdicate that the higher productive ratio for human rIL-6 inducing expression was obtainde in E. coli DH5a at O. D=0. 7. 5?10~6 unites of human rIL-6 HPGF bioactivities were produced from 1 litre of bacteria extracts assayed by 3HTdR uptake of 7TD1 cell line.

To find novel antihepatitis drugs, a series of nitrate-oleanolic acid (OA) hybrids (10a, 10b, 11a-11e and 12a-12c) were designed and synthesized on the basis of previous studies using OA as lead compound, which is widely found in natural plants and liver-specific metabolism. In the present study, ten novel NO-releasing derivatives of OA were synthesized by connecting nitrate to the OA-3-OH through varying lengths of linkers containing antioxidants which were designed to increase the ability of these target compounds to scavenge free radicals. The structures of these objective compounds were determined by IR, MS, 1H NMR and elemental analysis. Their protective effects on anti-Fas mediated HepG2 cell apoptosis were in vitro evaluated by LDH assay. Compound 12a is the most potent inhibitor. Its effect on anti-Fas mediated HepG2 cell apoptosis and amount of NO-releasing in vitro are similar to those of positive control NCX-1000.

Objective To analyze the age difference of gram negative isolates(GNB)from blood culture and antibiotic resistance profile between children and adults. Methods Age difference of pathogen distribution of GNB isolated from blood culture during 2015-2017 were retrospectively analyzed. WHONET 5.6 and Graphpad Prism 6 were used to perform Chi-square test on the pathogen proportion and antibiotic resistance rate. Results A total of 20 145 bacterial strains were isolated in Chongqing from 2015 to 2017. The top three strains of GNB were E. coli(56.7％, 6 688/11 799), K. pneumoniae(19.6％, 2 308/11 799), and P. aeruginosa(4.4％, 522/11 799). The resistant rate of E. coli to carbapenems was less than 1％. The resistant rate of K. pneumoniae to carbapenems was about 5％, while the resistant rate of S. marcescens was approximately 9％, similar to E. cloacae. The overall resistance rates of E. coli from adult patients to most antiobiotic agents were significantly higher than those of children, but the overall resistance rate of K. pneumoniae from children was higher than those of adults. The drug resistance rate of A. baumannii was higher than P. aeruginosa. A. baumannii isolates from adult patients were highly resistant to all drugs tested and more resistant than those from pediatric patients. Conclusions Majority of GNB strains isolated from blood culture were E. coli and K. pneumoniae, for which carbapenems are still active. More attention should be paid to carbapenem-resistant K. pneumoniae from children and A. baumannii from adults. National surveillance of nosocomial bloodstream infection should be highly evaluated.

Objective: To evaluate the efficacy and safety of reduced volume hepatectomy in treatment of advanced hepatic alveolar echinococcosis. Methods: The clinical data of 90 patients with advanced hepatic alveolar echinococcosis treated at the Qinghai Provincial People's Hospital from January 2017 to January 2019 were retrospectively analyzed. There were 41 males and 49 females, with an average age of 32 (range 11 to 58) years. The locations of the lesions, operations, complications and follow-up were analyzed. Results: 90 patients were treated with reduced volume focal hepatectomy, 38 with radical resection and 52 with quasi radical resection. The operation time was (361±22) min. The hospital stay was (22±2) day, and the blood loss was (781±37) ml. Red blood cells were transfused in 19 patients and plasma in 39 patients. Pringle’s maneuver was used in 12 patients, regional hepatic blood flow occlusion in 42 patients, and Glisson sheath occlusion in 26 patients. The total bilirubin, alanine aminotransferase and aspartate aminotransferase returned to normal in 3 to 14 days after operation. There were 12 patients who developed bile leakage, 41 pleural effusion and 26 effusion in the operation sites. A total of ninety patients were followed up for 2 to 24 months. There was no recurrence of echinococcosis after radical resection and no enlargement of residual lesions after quasi-radical resection. Conclusion Reduced-volume hepatectomy reduced the risk and difficulty of operation. The follow-up results were good. This approach provides a feasible scheme for treatment of advanced hepatic alveolar echinococcosis.

Objective: To compare the changes in liver function in patients with different types of hepatic alveolar echinococcosis after radiofrequency ablation. Methods: The data of 32 patients with hepatic alveolar echinococcosis treated by radiofrequency ablation from December 2016 to December 2018 at the Qinghai Provincial People's Hospital were retrospectively analyzed. There were 12 males and 20 females. The patients were divided into the single lesion group (n=17) and the multiple lesions group (n=15) according to the number of hydatid lesions. The lesions were further divided into the small lesion group (n=12) and the large or medium lesion group (n=20). The operation time, changes in AST and ALT were compared among these groups. Results: The operation time of the single lesion group was 1.5 (1.0, 1.8) hour, and that of the multiple lesions group was 3.0 (1.5, 3.5) hour. The difference was statistically significant (P<0.05). On the 1st and 3rd day after treatment, ALT in the multiple lesions group was significantly higher than that in the single lesion group (P<0.05). Compared with the 1st day after operation, ALT of the two groups decreased on the 5th day after treatment, and the difference was statistically significant (P<0.05). The changes in AST in the two groups were basically the same as that of ALT. The operation time of the small lesion group was 1.0 (1.0, 1.9) hour, and of the large and medium lesion group was 2.5 (1.5, 3.0) hour. The difference was statistically significant (P<0.05). On the 1st, 3rd and 5th day after treatment, ALT in the small lesion group was significantly lower than the large or medium lesion group (P<0.05). Compared with the 1st day after treatment, ALT of the two groups decreased on the 5th day after treatment, and the difference was statistically significant (P<0.05). There was a significant difference in AST between the small lesion group and the large or medium lesion group only on the 1st day after operation (P<0.05). Conclusion Radiofrequency ablation for hepatic alveolar echinococcosis with a single lesion and a small lesion had shorter operation time with less changes in liver function after treatment.

Objective To investigate the postoperative complications of ex vivo liver resection combined with autologous liver transplantation in the treatment of end-stage hepatic alveolar echinococcosis at high altitude and related prevention and treatment strategies. Methods Surgical data and follow-up data were collected from 11 patients with end-stage hepatic alveolar echinococcosis who underwent autologous liver transplantation in Qinghai People's Hospital from January 2013 to March 2019, and intraoperative and postoperative conditions were analyzed. Results All 11 patients underwent autologous liver transplantation successfully, without intraoperative death, among whom 2(18.18%) underwent hemi-extracorporeal hepatectomy and 9 (81.82%) underwent total extracorporeal hepatectomy. For the reconstruction of the retrohepatic inferior vena cava, 2 patients (18.18%) underwent reconstruction with the autologous great saphenous vein, 4 patients (36.36%) underwent reconstruction with artificial vessels, and the autologous retrohepatic inferior vena cava was preserved in 5 patients (45.45%). For biliary reconstruction, 8 patients (72.73%) underwent choledochoenterostomy and 3 (27.27%) underwent choledochocholedochostomy. The main postoperative complications of the 11 patients included bleeding in 2 patients (18.18%), bile leakage and abdominal infection in 4 patients (36.36%), bilioenteric anastomotic stenosis in 1 patient (9.09%), thrombus in 2 patients (18.18%), pulmonary infection and pleural effusion in 2 patients (18.18%), and echinococcosis recurrence in 1 patient (9.09%). Of all 11 patients, 2 (18.18%) died during the perioperative period, and the other 9 patients (81.82%) were improved and discharged. Conclusion Bleeding, biliary complications, and infection are the main causes of death in patients undergoing autologous liver transplantation at high altitude. An accurate understanding of surgical indication, careful multidisciplinary evaluation before surgery, superb operation during surgery, standardized surgical procedures, and fine perioperative management are the key to reducing perioperative mortality, avoiding and reducing postoperative complications, and achieving good long-term survival in patients undergoing autologous liver transplantation.