Genistein and its monoglucoside derivatives play important roles in food and pharmaceuticals fields, whereas their applications are limited by the low water solubility. Glycosylation is regarded as one of the effective approaches to improve water solubility. In this paper, the glycosylation of sophoricoside (genistein monoglucoside) was investigated using a cyclodextrin glucosyltransferase from Penibacillus macerans (PmCGTase). Saturation mutagenesis of D182 from PmCGTase was carried out. Compared with the wild-type (WT), the variant D182C showed a 13.42% higher conversion ratio. Moreover, the main products sophoricoside monoglucoside, sophoricoside diglucoside, and sophoricoside triglucoside of the variant D182C increased by 39.35%, 56.05% and 64.81% compared with that of the WT, respectively. Enzymatic characterization showed that the enzyme activities (cyclization, hydrolysis, disproportionation) of the variant D182C were higher than that of the WT, and the optimal pH and temperature of the variant D182C were 6 and 40℃, respectively. Kinetics analysis showed the variant D182C has a lower K_m value and a higher k_cat/K_m value than that of the WT, indicating the variant D182C has enhanced affinity to substrate. Structure modeling and docking analysis demonstrated that the improved glycosylation efficiency of the variant D182C may be attributed to the increased interactions between residues and substrate.
Cyclodextrins ; Genistein ; Glucosyltransferases/metabolism* ; Glycosylation ; Kinetics

Objective    To summarize the experience of totally thoracoscopic cardiac surgery for ventricular septal defect. Methods    Clinical data of 449 patients undergoing totally thoracoscopic cardiac surgery for ventricular septal defect from May 2008 to December 2018 in Shanghai Yodak Cardiothoracic Hospital were analyzed retrospectively. There were 232 male and 217 female patients, aged from 3 to 55 years with a mean age of 17.3±11.2 years. Results    All the operations were completed successfully. Mean operative time was 2.4±0.3 h. The mean extracorporeal circulation time and aortic cross-clamp time was 64.2±11.6 min and 28.4±10.7 min, respectively. Mechanical ventilation time and intensive care unit stay was 6.9±3.8 h and 20.5±5.6 h, respectively. Postoperation drainage quantity was 213.1±117.2 mL. The hospital stay was 6.9±1.3 d. Intraoperative and postoperative complications occurred in 11 patients (2.4%), including 1 patient of intraoperative reoperation, 3 patients of reoperation for bleeding, 3 patients of the incision infection, 2 patients of small residual shunt, 1 patient of right femoral artery incision stenosis complicated by thromboembolism and 1 patient of right pleural cavity pneumothorax. The mean follow-up time was 72.2±33.9 months. During the period, there was no reoperation, but 2 patients of ventricular septal defect small residual shunt, 1 patient of mild-moderate mitral valve and 1 patient of mild-moderate aortic valve incompetence, respectively. During the period, heart function of the patients was NYHAⅠ-Ⅱ. Conclusion    Totally thoracoscopic cardiac surgery for ventricular septal defect is a safe and effective treatment, with few serious complications, fast recovery for patients and good short to medium-term outcomes.

Objective    To summarize the experience of totally thoracoscopic cardiac surgery (TTCS) for atrial septal defect. Methods    Clinical data of 442 patients undergoing TTCS for atrial septal defect from May 2008 to December 2018 in Shanghai Yodak Cardiothoracic Hospital was analyzed retrospectively. There were 149 male and 293 female patients, aged 3-74 (29.1±14.3) years. Surgical procedures were performed through 3 ports at the right chest wall. Results    All the operations were completed successfully. Mean operative time was 1.5-4.6 (2.2±0.3) h. The mean extracorporeal circulation and aortic cross-clamp time was 28-118 (55.9±13.3) min and 8-78 (21.5±10.2) min, respectively. Mechanical ventilation and intensive care unit stay time was 3.5-122.0 (8.1±7.4) h and 13-141 (20.7±10.2) h, respectively. Postoperation drainage volume was 70-1 280 (251.8±131.5) mL. The hospital stay was 4-16 (7.1±1.4) d. Intraoperative and postoperative complications occurred in 15 patients (3.3%). The mean follow-up time was 1-128 (67.6±33.3) months, and during the period, there were 25 patients of atrial fibrillation, 25 patients of mild-moderate tricuspid valve incompetence, 1 patient of moderate tricuspid valve incompetence. There was no reoperation or residual shunt during the period of follow-up. And the heart function was improved. Conclusion    TTCS is a feasible, safe and minimal invasive approach for patients with atrial septal defect and has good short to medium-term outcomes.