Objective To study the effect of repairing limbs bone defect by using uncellular tissue engineering complex of autolegous red bone marrow wrapped by facial flap with vessels to provide evidence for clinical application. Methods Nineteen cases of limbs bone defect were chosen, among them, 3 cases were with benign bone tumor,6 cases were with open fracture causing bone defect and 10 cases were with bone hypotoxic infection after operation. Took autolegous red bone marrow to prepare uncelluar tissue engineering complex with osteoinduction active material (OAM) containing bone morphogenetic protein (BMP). Prepared a facial flap with capillary network originating from an anonymos vessel adjacent to the bone defect,wrappad the tissue engineering bone and filled the bone defect. On a certain time after the operation, the patients were tested by X-ray. Results All the 19 cases were followed up in 1st,3rd,6th, 10th, 12th and 18th month after the operation. There were 3 cases that the implanted tissue engineering bone completely replaced the bone defect in 6th month displayed on X-ray,6 cases in 10th month,9 cases in 12th month and 1 case in 18th month. None of the cases had bone infection or bone absorption. Osteanagenesis and bone molding had come true in all cases. Conclusions The uncellular tissue engineering complex of autolegous red bone marrow wrapped by facial flap with vessels shows double effects in inducing osteanagenesis and the vascularization, and it is feasible in the recovery of large area bone defect. The complex can promote bone recovery and advance the quality and quantity of osteanagenesis.

A total of 38 cases of senile brucella spondylitis disease at our hospital during January 2002 and March 2012 were analyzed .After admission , all of them were definitely diagnosed on the basis of epidemiological history , clinical manifestations , laboratory tests , imaging and pathological examinations . Over a follow-up period of 12 months, 17 cases were cured after standardized drug treatment .Among 21 surgical cases, there were curing (n=17) and improving (n=2).Senile brucellosis spondylitis has distinct serological and pathological characteristics .And formulating the diagnostic criteria may improve its diagnostic rate and reduce its misdiagnostic rate .And standardized drug therapy achieves a better curing rate and a proper timing of surgical intervention improves its clinical outcomes .

Objective To study the effect of the uncellular tissue engineering complexes of autolegous red bone marrow wrapped by facial flap with vessels in repair of large segment bone defect infected with low virulence bacteria so as to provide evidence for the clinical application. Methods The study included 38 cases of limb bone defect infected with low virulence bacteria after trauma.Autologous red bone marrow (ARBM) was taken to prepare uncelluar tissue-engineered complexes with osteoinductive absorbing material (OAM) containing bone morphogenetic protein (BMP).A facial flap with capillary network originating from an anonymous vessel adjacent to the bone defect was prepared to wrap the tissue engineered bone and fill the bone defect.Pathological focus clearance and tissue-engineered complexes compounded with ARBM implantation were performed in 18 cases (Group A) and pathological focus clearance and tissue-engineered complexes of autolegous red bone marrow wrapped by facial flap with vessels implantation in the other 20 cases ( Group B).The blood routine and supersensitive CRP were examined to monitor the inflammation reaction; X-ray was used to observe the bone defect repair; histology and bacteriology examinations were performed in partial cases at 3,6,12,18 months after operation. Results Six months after operation,5 cases of Group A were infected and the bacteria cultivation was as positive as that before the operation.The histological observation at ( 14.0 ± 0.5 ) months after operation showed that fibrous connective tissues between the bone fracture ends existed in the pathological area in 10 cases,of whom four cases were filled with inflammatory fibrous granulation tissues and few dead bones in the pathological area,and the bacterial examination was positive.There was no infection in Group B after operation.The histological observation manifested periosteum like tissues formation from the primary facial flap,mature bone structure formation in the primary pathological area and non-inflammatory infiltration in 16 cases and the bacteria cultivation was negative in these cases.The external fixation frame was taken out (12.2 ± 0.3 )months after operation because the synostosis appeared and the structure was stable in the other seven cases including three cases in Group A and four in Group B and the histological and bacterial examination were not performed.At each time point after operation,not only the blood routine but also the supersensitive CRP and the X-ray quantification grade of Group B were significantly more than those of Group A (P ＜ 0.05 ). Conclusions The uncellular tissue-engineered complexes of autolegous red bone marrow wrapped by facial flap with vessels is a feasible method for repairing the infected bone defect by first intention,since it can resist infection,obviously promote the bone recovery and advance the quality and quantity of osteanagenesis.

Objective To investigate the prognostic value of C5L2 in patients with hepatocellular carcinoma (HCC).Methods The data of 175 patients with HCC who underwent curative resection at Zhongshan Hospital,Fudan University from Oct.,2012 to Sep.,2013 were analyzed retrospectively.The correlation between C5L2 and clinicopathologic characteristics were explored.COX regression model was used to determine the influence of clinical parameters on predicting recurrence,and the prognostic value of combined application of C5L2 and AFP were evaluated by Kaplan-Meier method.In vitro,the expression of C5L2 were tested in 5 HCC cell lines,and Hep3B and Huh7 were chosen for down-regulation and up-regulation of C5L2,respectively,the abilities of invasion and migration were examined by transwell and the potential mechanism was explored.Results The C5L2 expression was correlated to gender,tumor size and recurrence,and the recurrence rate of low C5L2 expression group was higher.Also,the multivariate analysis showed that C5L2 low expression was an independent risk factor for recurrence.Moreover,the combined application of C5L2 and AFP could estimate prognosis more effectively.Knockdown of C5L2 in Hep3B promoted the invasiveness and motility,and increased the level of β-catenin and MMP2;conversely,overexpression of C5L2 in Huh7 inhibited the invasiveness and motility,and decreased the level of β catenin and MMP2.Conclusions C5L2 could be regarded as an auxiliary indicator for prognosis of HCC,thereby the evaluation of C5L2 could help with making effective and comprehensive management for HCC patients.

Objective:To verify the performance of the next-generation sequencing (NGS) platform and evaluate the application of NGS, droplet digital PCR (ddPCR) and super amplification refractory mutation system (super-ARMS) in the detection of circulating free DNA (cfDNA) mutations in patients with non-small-cell lung cancer (NSCLC).Methods:A total of 75 patients with NSCLC in the respiratory department of Zhongshan Hospital Affiliated to Fudan University were enrolled. The standards, cfDNA from 25 patients with newly diagnosed and untreated NSCLC, and self-made mixed samples mixed with hemoglobin (1 000 mg /dl), bilirubin (500 mg/l), fat emulsion (2%), enterococcus gDNA and Escherichia coli gDNA were used to verify the blank limit, analytical sensitivity, precision, accuracy and specificity of NGS platform. The cfDNA mutations of 75 NSCLC patients were detected by ddPCR and NGS, and the mutation positive rates of the two platforms were compared. The linear relationship between the two platforms was compared by Pearson correlation test. 12 patients were selected by simple random sampling for the detection of plasma super-ARMS platform. The performance of three platforms in the detection of plasma cfDNA mutation in patients with NSCLC was compared.Results:The blank limit of NGS platform was set to 0.00%, the analytical sensitivity was 0.2%, the intra-assay precision and inter-assay precision were 100%. The test results were not affected by endogenous hemoglobin, bilirubin or fat emulsion in plasma or exogenous DNA interference, and the analysis specificity was good. The mutation positive rates of plasma cfDNA in 75 NSCLC patients detected by ddPCR and NGS were 61.33% and 60.00%, respectively. The complete coincidence rate was 89.33%, which suggests there was a positive correlation between the mutation abundance of NGS and ddPCR ( r=0.984, P=0.001). Among the plasma of 12 NSCLC patients, the results of NGS, ddPCR and super-ARMS were completely consistent in 7 cases, including 2 wild-types and 5 mutants. Conclusion:The NGS platform was verified to be useful for cfDNA mutation detection in patients with NSCLC. The ddPCR, NGS and super-ARMS have their own advantages in detecting cfDNA mutations in patients with NSCLC.

Objective To investigate the effect of dapagliflozin on metabolic markers, hepatic fat content, and autonomic nervous function in patients with type 2 diabetes mellitus (T2DM) and metabolic associated fatty liver disease (MAFLD). Methods A total of 90 patients with T2DM and MAFLD who were admitted to The Second Affiliated Hospital of Zhengzhou University from October 2019 to October 2020 were enrolled and randomly divided into control group and dapagliflozin group, with 45 patients in each group. All patients were given conventional treatment before enrollment; the patients in the control group were treated with the original hypoglycemic regimen, and those in the dapagliflozin group were given dapagliflozin in addition to the treatment in the control group. The treatment cycle was 24 weeks. General information was collected before and after treatment, and the two groups were compared in terms of the changes in body mass index (BMI), glycosylated hemoglobin (HbA1c), fasting blood glucose (FPG), blood lipids, serum uric acid (SUA), Homeostasis Model Assessment of Insulin Resistance (HOMA-IR), liver function, liver fat content, and heart rate variability after treatment. The paired t -test was used for comparison of normally distributed continuous data within each group, and the independent samples t -test was used for comparison between groups; the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data within each group, and the Mann-Whitney U test was used for comparison between groups. The Chi-square test was used for comparison of categorical data between two groups. Results A total of 43 patients in the dapagliflozin group and 40 patients in the control group completed the study. After 24 weeks of treatment, the dapagliflozin group had significant reductions in BMI, HbA1c, FBG, triglyceride (TG), SUA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), HOMA-IR, and liver fat content ( t =8.781, 8.765, 8.813, 3.485, 6.199, 5.694, 3.428, 6.492, and 4.925, all P < 0.05) and significant increases in high-density lipoprotein cholesterol, standard deviation of all normal R-R intervals (SDNN), standard deviation of average NN intervals (SDANN), root mean square of successive differences, percent of the number whose difference between adjacent NN interval are more than 50 ms (pNN50), high frequency (HF), and low frequency (LF) ( t =-2.055, -6.307, -7.696, -3.388, and -7.928, Z =-3.339 and -3.309, all P < 0.05), while the control group had significant reductions in HbA1c, FBG, and HOMA-IR ( t =9.220, 7.214, and 3.340, all P < 0.05). Compared with the control group after treatment, the dapagliflozin group had significantly lower levels of BMI, HbA1c, TG, SUA, HOMA-IR, ALT, AST, and liver fat content ( t =-4.055, -2.670, -2.056, -2.496, -3.976, -3.703, -2.123, and -5.184, all P < 0.05) and significantly higher levels of SDNN, SDANN, pNN50, LF, and HF ( t =4.136, 5.433, and 5.971, Z =-2.333 and -2.010, all P < 0.05). Conclusion For patients with T2DM and MAFLD, dapagliflozin can reduce BMI, HbA1c, TG, SUA, and liver fat content, improve insulin resistance and liver function, reduce the activity of sympathetic nerve, and regulate autonomic nerve function.