To obtain fusion protein of Mycobacterium bovis with high purity, the recombinant prokaryotic expression vector for Mb2277 gene was constructed and the immunogenicity of its products was initially investigated in the present study.A pair of primer was designed according the gene sequence Mb2277 from the genomic DNA of M.Bovis in GenBank. and was amplified by PCR using DNA of M.Bovis 93006 strain as template. The PCR product and pET-28a(+) was then digested by BamHⅠ and EcoR Ⅰdouble enzyme. To constructed a prokaryotic expression plasmid, the purified Mb2277 was cloned to pET28a(+). Then the recombinant plasmid was transformed into competent cell of E.coli BL21(DE3).The bacteria were induced by IPTG and its lysates were analyzed by SDS-PAGE and Western-blotting. In this way, the prokaryotic expression plasmid for M. bovis Mb 2277 protin was obtained, and a expression band with molecular of 25 ku could be found in SDS-PAGE analysis. As demonstrated by Western blotting this expression product showed excellent reactivity with rabbit immune sera against M. bovis.

Objective To observe the clinical efficacy of vacuum sealing drainage(VSD) in the treatment of large-area wound infection of earthquake casualty after amputation.Methods Seven patients with large-area wound infection of earthquake casualty after amputation received sustained VSD.Results After VSD for 7~26 days,with an average of 13 days,the wound infection in the 7 patients was controlled.No systemic toxicity was found.The result of wound bacterial culture was negative.Of the 7 patients,4 received phase Ⅱ suture,3 received phase Ⅱ skin graft,and all of the grafted skin survived.Conclusion Vacuum sealing drainage exerts certain effect in the treatment of large-area wound infection of earthquake casualty after amputation.

Objective To investigate the effect of total flavones of Rhizoma Drynariae on femur distraction osteogenesis in the rabbits. Methods Thirty-two healthy rabbits were randomly divided into treatment group and control group, 16 rabbits in each group. The femoral fracture was treated with unilateral femoral distraction and was fixed with a self-made distraction instrument. After 7-day intermittent period, the fractured femur was distracted at a rate of 1 mm/d, twice a day for 10 continuous days. The treatment group was fed with total flavones of Rhizoma Drynariae from the first post-operative day to the end of the experiment. And then all of the animals were sacrificed after fixation for 28 days. The bone specimens were used for histological observation and immunohistochemical detection. Results The area of mature bone in the newborn bone tissue of the treatment group was increased, and osteoblasts number and the percentage of trabecular bone area were significantly higher than those of the control group . The bone morphogenetic protein-2 (BMP-2) and transforming growth factor-β1 (TGF-β1) were stained brown deeply, the staining degree being stronger than that of the control group. Conclusion Rhizoma Drynariae total flavones can effectively accelerate the formation and maturation of newborn bone tissue during bone distraction.

BACKGROUND: Ankle fracture combined with deltoid ligament rupture and distal tibiofibular syndesmosis injury occurs occasionally. Its treatment with distal tibiofibular syndesmosis screw fixation or deltoid ligament repair remains controversial. The former appears with poor reduction, broken nails, secondary surgery and other problems.OBJECTIVE: To observe the clinical effectiveness of suture anchor repair for ankle fracture combined with deltoid ligament injury.METHODS: Twelve patients with ankle fracture combined with deltoid ligament injury were selected from the First Affiliated Hospital of Guangzhou University of Chinese Medicine between January 2013 and December 2015. All patients were treated with open reduction, internal fixation, and anchor repair, but without distal tibiofibular syndesmosis screw fixation. The curative efficacy and joint stability were observed.RESULTS AND CONCLUSION: (1) All patients were followed-up for more than 12 months. (2) The modified Baird-Jackson scoring system showed excellent in nine cases, good in two cases, average in one case, poor in none case, and the excellent and good rate was 92%. (3) At 1 year after internal fixation, the X-ray examination showed the malleolus gap and talus slope angle of the affect side were (3.47±0.12) mm and (0.45±0.18)°, and the malleolus gap and talus slope angle of the healthy side were (3.44±0.05) mm and (0.43±0.14)°, and there was no significant difference between two sides (P > 0.05). (4) These results indicate that the suture anchor can repair the anatomy and biomechanics of deltoid ligament with stable ankle joint, and secondary surgery is unnecessary.

Objective To evaluate laparoscopic balloon nasobiliary biliary drainage (LBNBD),vs ureteral catheter drainage in one stage laparoscopy,choledochoscopy and duodenoscopy choledocholithotomy and primary closure of the small calibered common bile duct (diameter 0.3-0.8 cm).Methods During the period of Apr 2010 to Nov 2016 102 cases were enrolled including 50 cases receiving LBNBD and 52 cases using ureteral catheter drainage.Results Between the two groups,LBNBD was superior to ureteral catheter drainage in all the following parameters:the operation time,intraoperative blood loss,postoperative liver function,blood amylase and other laboratory indicators,gastrointestinal function recovery time,gastrointestinal symptoms and electrolyte imbalance,postoperative hospital stay,and bile duct drainage time with all differences statistically significant (P ＜ 0.05).Bile drainage differences during the postoperative first 3 days (averagely 200-400 ml a day) were not statistically different (P ＞ 0.05).Postoperative pancreatitis,bile leakage,and hemobilia were not statistically different (P ＞ 0.05).Conclusions The use of LBNBD is safe and effective in endoscopic choledocholithotomy in cases of small calibered common bile duct.

Objective:To analyze the influencing factors of death in patients with acute respiratory distress syndrome (ARDS) combined with acute kidney injury (AKI) in intensive care unit (ICU) after continuous renal replacement therapy (CRRT).Methods:The demographic and clinical data of ICU patients with ARDS combined with AKI after CRRT at Henan Provincial People's Hospital from January 1, 2018 to December 31, 2018 were collected. According to the final treatment results of this hospitalization, the patients were divided into death group and survival group. Survival was defined as the improved patient's condition and hospital discharge. Death was defined as the patient's death during the ICU hospitalization or confirmed death after abandoning treatment and automatically being discharged from the hospital in the follow-up. The basic clinical characteristics and CRRT status between the two groups were compared. Multivariate logistic regression method was used to analyze the influencing factors of death in patients.Results:A total of 132 patients were enrolled, of which 90 patients (68.2%) died, with 84 males (63.6%) and median age of 59(45, 73) years. Compared with the survival group, the death group had higher age, proportion of malignant tumors, sequential organ failure assessment (SOFA) score, number of organ dysfunction and proportion of positive balance of fluid accumulation at 72 hours, longer time from entering ICU to CRRT, and lower mean arterial pressure (minimum value) and oxygenation index (all P<0.05). Multivariate logistic regression analysis results showed that the age≥60 years old ( OR=4.382, 95% CI 1.543-12.440, P=0.006), large number of organ dysfunction ( OR=1.863, 95% CI 1.109-3.130, P=0.019), high SOFA score ( OR=1.231, 95% CI 1.067-1.420, P=0.004) and long time from ICU admission to CRRT ( OR=1.224, 95% CI 1.033~1.451, P=0.020) were independent influencing factors of death in patients with ARDS combined with AKI after CRRT, and high oxygenation index ( OR=0.992, 95% CI 0.986-0.998, P=0.010) was an independent protective factor for patients' prognosis. Conclusions:The mortality of patients with ARDS combined with AKI after CRRT is still high. The age≥60 years old, large number of organ dysfunction, high SOFA score and long time from ICU admission to CRRT are independent influencing factors for death, and high oxygenation index is an independent protective factor for prognosis in patients with ARDS combined with AKI after CRRT.

Objective:To investigate the incidence of acute kidney injury (AKI) following pancreaticoduodenectomy and related risk factors in elderly patients.Methods:The clinical data of elderly patients who underwent pancreaticoduodenectomy in Henan Provincial People′s Hospital from January 2017 to June 2020 were collected retrospectively. According to the changes of serum creatinine within 48 h or 7 days after operation, the patients were divided into AKI group and non-AKI group. The basic clinical characteristics of the two groups were compared, and the incidence of AKI was calculated. Multivariate logistic regression model was used to analyze the risk factors of postoperative AKI.Results:A total of 322 elderly patients were enrolled, with age of (67.1±5.2) years old (60-85 years old) and 186 males (57.76%). Among 322 elderly patients, there were 41 patients (12.73%) suffering from AKI following pancreaticoduodenectomy. Compared with the non-AKI group, the level of bilirubin in AKI group was higher ( Z=-2.012, P=0.044), and the level of hemoglobin in AKI group was lower ( Z=-2.111, P=0.035). Multivariate logistic regression analysis showed that increased preoperative total bilirubin ( OR=1.003, 95% CI 1.000-1.006, P=0.027) and postoperative exploratory laparotomy ( OR=3.936, 95% CI 1.071-14.460, P=0.039) were the independent influencing factors for AKI after pancreaticoduodenectomy in elderly patients. Conclusions:The incidence of AKI after pancreaticoduodenectomy in elderly patients is 12.73%. Preoperative high bilirubin and postoperative exploratory laparotomy may be the independent risk factors for AKI after pancreaticoduodenectomy in elderly patients.

BACKGROUND:Studies have shown that both Panax notoginseng saponins and concentrated growth factor can promote fracture healing,but there are few studies addressing their combined effects on fracture healing.Panax notoginseng saponins may accelerate fracture healing by promoting the release of concentrated growth factor-related factors over a certain period of time. OBJECTIVE:To study the effect of Panax notoginseng saponins on concentrated growth factor release and fracture healing in rats. METHODS:Eighteen 8-week-old Sprague-Dawley rats were numbered and randomly divided into three groups:Panax notoginseng saponins group,model control group and blank group.Panax notoginseng saponins group was fed with Panax notoginseng saponins for 2 weeks.Model control group was given 2 mL of normal saline for 2 weeks and blank group was fed normally.Concentrated growth factor was obtained by the centrifugation method both from the Panax notoginseng saponins group and model control group.After 1 week of normal feeding,all animals underwent modeling for femoral fracture.The Panax notoginseng saponins group and the model control group were implanted with autologous concentrated growth factor,and then the release concentration of growth factors at different time points(1 hour,1,3,5,7,9 and 11 days)were measured by ELISA.Fracture healing was assessed based on postoperative X-ray and hematoxylin-eosin staining of bone tissues. RESULTS AND CONCLUSION:Compared with the model control group,the Panax notoginseng saponins group had higher release concentrations of vascular endothelial growth factor A and transforming growth factor β at 7,9,and 11 days,Platelet-derived growth factor BB at 5,9,and 11 days,and basic fibroblast growth factor at 1-11 days(P<0.01).X-ray examinations indicated that fracture healing in the Panax notoginseng saponins group was better than that in the model control group,and fracture healing in these two groups was better than that in the blank group at 2 months after surgery.Hematoxylin-eosin staining results found that the constituent osteocyte density in the Panax notoginseng saponins group was greater than that in the model control group,and the constituent osteocyte density in these two groups was better than that in the blank group.These findings indicate that Panax notoginseng saponins can increase the concentration of concentrated growth factor-related factors.After intervention with Panax notoginseng saponins,concentrated growth factors are more advantageous in promoting fracture healing in rats.

Objective To estimate the dose-response relationship between sedentary behavior with mortality in patients with type 2 diabetes. Methods A total of 17786 type 2 diabetic patients were recruited as participants, who were included in National Basic Public Health Service in Changshu County of Suzhou City, Qinghe District and Huai'an District in Huai'an City of Jiangsu Province. Cox proportional hazards regression model and restricted cubic spline model were employed to estimate the dose-response relationship between sedentary behavior with all-cause and cause specific mortality in patients with type 2 diabetes. Results Among 78114.34 person-years of the fo1low-up, the median of follow-up time was 4 years, and 1285 deaths occurred during that period. Compared to patients with sedentary behavior≤2 h/d, the multivariate adjusted hazard ratios of all-cause death associated with sedentary behavior levels of 3-4 h/d, 5-6 h/d, and≥7 h/d were 1.05(95％CI 0.92-1.20), 1.20(95％CI 1.03-1.42), and 1.39 (95％CI 1.16-1.65), respectively. Eevry increase of 1 h/d in sedentary behavior was associated with an increased hazard of death from cardiovascular disease(CVD) of 4％(HR=1.04, 95％CI 1.01-1.07) and from other causes of 6％( HR=1.06, 95％CI 1.03-1.09) . However, no significant association between sedentary behavior and malignant tumor death was found. The multivariable restrictive cubic spline regression indicated that the linear dose-response relationships were found between sedentary time with the all-cause, CVD cause, and other cause of mortality ( Non-linear test, P>0.05) . Conclusion Longer sedentary behavior could increase the risk of mortality in patients with type 2 diabetes.

Objective:To predict the possible targets and signaling pathways of Jiangtang Xiaoke Granules in the treatment of diabetes mellitus (DM) using computer network pharmacology and molecular docking technology.Methods:The active components and targets of Jiangtang Xiaoke Granules were collected by ETCM; the targets of DM were searched from the databases of DisGeNET and GeneCards, and the intersections of the two were taken to draw a Venny diagram; String database was used for gene transformation and network interaction analysis; the network diagram was constructed with Cytoscape3.6.0; the predicted results were supported by molecular docking technology; GO and KEGG analysis was performed through Metascape database.Results:A total of 128 active components of Jiangtang Xiaoke Granules were screened, with 607 corresponding targets, 1 240 DM related targets, and 53 core targets. Molecular docking showed that the active components had good binding energy with the core targets. GO analysis yielded 46 functional items and KEGG analysis yielded 15 pathways.Conclusion:Jiangtang Xiaoke Granules regulate glucose homeostasis by participating in a variety of biological processes through multiple components, and multiple targets, including affecting lipids and atherosclerosis, Alzheimer disease, AMPK signaling pathway, Apelin signaling pathway, and glucagon signaling pathway.