1.Construction, expression, putification and bioactivity analysis of a two functional domains containing small molecule CR1 derivative
Yongtao YANG ; Li HE ; Gaoke LIU ; Bing TAN ; Zhengqing WANG
Chinese Journal of Microbiology and Immunology 2008;28(11):1044-1049
Objective To construct and express a small complement receptor type 1 (CR1) deriv-ative which contained two functional domains. Methods Total RNA was isolated from the peripheral blood mononuclear cell. The functional fragment Ⅰ of CR1 was amplified using the RT-PCR. The functional frag-ment Ⅱ was amplified with the plasmid of pET-32a-CR1-SCR15-18 as template which had been already con-structed in our laboratory. Then the chimeric gene that contained the two fragments was constructed with spli-cing overlap extention PCR. The chimeric gene was then inserted into the plasmid of pET-32a (+) and transformed into E. coli Rosetta(DE3). The inserted gene was verified by enzyme digestion and DNA se-quencing. After induced by IPTG, the expressed protein was analyzed by SDS-PAGE and Western blot. Then the fusion protein was purified by Ni-NTA affinity column and renatured through dialysis. The comple-ment inhibition activity was determined by CH50 method. Results The chimeric gene was successfully cloned into pET-32a(+). The result of SDS-PAGE and Western blot confirmed the expressed protein and showed that the molecule mass(Mr) of the expressed protein was 63×103. The purity of the recombinant protein was up to 92% after Ni-NTA column affinity chromatography. The bioactivity assay showed the fusion protein had a concentration-dependent complement inhibition activity within the concentration range of 0-200 μg/ml. Conclusion The two functional domains contained small CR1 derivative was successfully construc-ted and expressed in E. coli Rosetta. The fusion protein had a relative high bioactivity, providing a basis for further function experiment in vivo.
2.Absorbable artificial bone for repair of comminuted calcaneal fractures
Hui SUN ; Xuehui ZANG ; Lihua GAO ; Yongtao TAN
Chinese Journal of Tissue Engineering Research 2014;(39):6375-6380
BACKGROUND:Currently, surgical treatment for Sanders III type, IV type fractures to restore damaged subtalar surface of the calcaneus and calcaneal shape, in order to reduce the incidence of traumatic arthritis has reached a consensus, and whether bone grafting is selected during surgery for calcaneal fractures has been a controversial issue. OBJECTIVE:To observe the clinical effect of Genex synthetic bone transplants and locking plate on comminuted calcaneal fractures. METHODS:Twenty-one cases of Sanders III type, IV type calcaneal fractures were retrospectively analyzed, including 16 males and 5 females, aged 22 to 55 years. After fracture reduction, a dough-like Genex bone graft was implanted into the defect region via lateral“L”shaped approach, and then the lateral wal of the bone was reset fol owed by internal fixation with the pre-curved locking plate. Fol ow-up observation was performed for fracture healing, Bolher angle and the Maryland Foot Score. RESULTS AND CONCLUSION:Total y 21 patients were fol owed up for 8-16 months. The fractures were healed without displacement, col apse and rejection. The bone graft was degraded within 6 months and completed absorbed after 1 year. According to Maryland Foot Score, the excellent rate was up to 86%, and the Bolher angle was increased from an average preoperative (5.3±3.35)° to postoperative (24.3±1.06)°. Genex artificial bone meal is a biomaterial that can be completely absorbed, has good plasticity and strong supporting force, and it is able to ful y fil bone defects, be easy to fracture reduction, induce bone formation, and promote fracture healing.
3.Effect of different nutritional support modes on humoral immunity and outcomes after esophagectomy
Taichang TAN ; Changning YE ; Qiang FANG ; Guangguo REN ; Yongtao HAN ; Lin PENG
Chinese Journal of Clinical Nutrition 2011;19(6):372-376
ObjectiveTo explore the effect of different nutritional support mdoes on humoral immunity and outcomes after esophagectomy in patients with esophageal carcinoma.MethodsForty-six patients with middle or low thoracic esophageal carcinoma underwent Ivor Lewis esophagectomy.The patients were randomized into enteral nutrition group ( EN,n =23 ) and enteral combined parenteral nutrition group ( EN + PN,n =23 ) based on the nutrition support modes.Serum levels of immunoglobulin (IgG,IgA,IgM,IgE,κ/λ light chain) and comphments (C3/C4) were assayed and compared on the 1st pre-operative day and at 18 hours as well as 3rd and 7th day after operation.The clinical outcomes including infection-related complications and hospital stay were compared between two group s.ResultsThere was no significant difference in all humoral immunity indicators between two groups at the eachpost-operative time point.In both two groups,the levels ofIgG [ (8.90 ± 1.75),(7.53 ±1.41) g/Land (8.64±2.44),(7.48±2.16) g/L],κ [ (2.14±0.46),(1.78±0.41) g/L,and (2.15 ±0.63),( 1.86 ± 0.62) g/L] and λ light chain [ ( 1.34 ± 0.45 ),( 1.11 ± 0.31 ) g/L and ( 1.20 ± 0.32),( 1.08 ± 0.35 ) g/L] were significantly lower 18 hours and 3rd day after operation than the pre-operative levels [ (12.15±2.86)and (11.11±2.96) g/L,(2.90±0.77) and (2.77±0.79) g/L,(1.79±0.57) and (1.56±0.41) g/L] (P=0.000,P=0.000,and P=0.004,P=0.000,and P=0.000,P=0.000,and P=0.011,P=0.000,and P=0.004,P=0.000,and P =0.008,P =0.000),and returned to the preoperative levels by the postoperative 7th day (P>0.05),except for the level of κ light chain 7th day after operation in EN group [ ( 2.42 ± 0.69) g/L] ( P =0.027 ).The levels of IgA,IgE,and C3 were not significantly different during the perioperative period ( P > 0.05 ).The level of IgM was not significantly different during the perioperative period in EN group (P >0.05),and was significantly lower on the 3rd post-operative day [ ( 1.00 ±0.53) g/L] than the pre-operative level [ ( 1.47 ±0.76) g/L] in the EN + PN group (P =0.031 ),and were not significantly different on the other time points (P > 0.05 ).In the EN group,the C4 level was significantly lower at the postoperative 18 hours [ (0.24 ±0.08) g/L] than the pre-operative level [ (0.37 ±0.36) g/L] (P =0.030),and were not significantly different at the other time points ( P > 0.05 ).In the EN + PN group,the C4 level was not significantly different during the perioperative period ( P > 0.05 ).There was no significant difference in the infection-related complications and hospital stay between these two groups ( P =0.300,P =0.371 ).ConclusionsThe effects of EN or EN + PN on humoral immunity and outcomes after esophagectomy in patients with esophageal carcinoma are not different.Both these two nutritional support modes can not completely alleviate the harm to the humoral immunity.The EN is more cost-effective.
4.Chinese thoracic surgery experts consensus on postoperative follow-up plans for esophageal squamous cell carcinoma
Longqi CHEN ; Xiaofei LI ; Jianhua FU ; Song ZHAO ; Yin LI ; Yousheng MAO ; Shuoyan LIU ; Zhentao YU ; Lijie TAN ; Hui LI ; Yongtao HAN ; Chun CHEN ; Mingqiang KANG ; Jian HU ; Zhigang LI ; Hecheng LI ; Renquan ZHANG ; Shidong XU ; Linyou ZHANG ; Kaican CAI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(02):141-149
Resection is one of the most important treatments for esophageal squamous cell carcinoma, and routine postoperative follow-up is an effective method for early detection and treatment of recurrent metastases, which can improve patients' quality of life and prognosis. This consensus aims to provide a reference for colleagues responsible for postoperative follow-up of esophageal squamous cell carcinoma patients in China, and further improve the standardization of the diagnosis and treatment of esophageal squamous cell carcinoma.
5. Clinical effect and safety of pegylated interferon-α-2b injection (Y shape, 40 kD) in treatment of HBeAg-positive chronic hepatitis B patients
Fengqin HOU ; Yalin YIN ; Lingying ZENG ; Jia SHANG ; Guozhong GONG ; Chen PAN ; Mingxiang ZHANG ; Chibiao YIN ; Qing XIE ; Yanzhong PENG ; Shijun CHEN ; Qing MAO ; Yongping CHEN ; Qianguo MAO ; Dazhi ZHANG ; Tao HAN ; Maorong WANG ; Wei ZHAO ; Jiajun LIU ; Ying HAN ; Longfeng ZHAO ; Guanghan LUO ; Jiming ZHANG ; Jie PENG ; Deming TAN ; Zhiwei LI ; Hong TANG ; Hao WANG ; Yuexin ZHANG ; Jun LI ; Lunli ZHANG ; Liang CHEN ; Jidong JIA ; Chengwei CHEN ; Zhen ZHEN ; Baosen LI ; Junqi NIU ; Qinghua MENG ; Hong YUAN ; Yongtao SUN ; Shuchen LI ; Jifang SHENG ; Jun CHENG ; Li SUN ; Guiqiang WANG
Chinese Journal of Hepatology 2017;25(8):589-596
Objective:
To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 μg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control.
Methods:
This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (