Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.

Objective:To investigate the clinical effect of laparoscopic assisted removal of greater omentum free transplantation combined with skin grafting for the repair of large area refractory wounds.Methods:From June 2013 to June 2018, 18 cases of lower extremity skin and soft tissue defects with multiple bone, joint, tendon and internal plants exposure were admitted to Hanzhong Central Hospital, including 12 males and 6 females, aged from 15 to 50 years old, with an average age of 32.6 years old. The area of skin and soft tissue defect: 30 cm×12 cm-53 cm×21 cm. The operation was divided into two stages. In the first stage, the greater omentum was acquired with the assist of laparoscope and free transplanted to cover the wound. After the greater omentum free transplantation was confirmed to survive, the split-thickness skin graft was applied for wound repair.Postoperative survival of the greater omentum and skin grafting, complications, appearance and function of lower limbs were observed and followed up.Results:The 18 operations were performed successfully, the area of omentum resection was 25 cm×10 cm-35 cm×15 cm, all the greater omentums survived after operation without complications such as intestinal adhesion, volvulus and peritonitis. The area of the skin grafting was 36 cm×8 cm-45 cm×22 cm. 16 cases skin grafting survived completely, 2 cases skin grafting were necrosis just local small area, and scar healed after dressing change. Postoperative follow-up of 6-12 months showed good appearance and function of lower limbs and satisfactory results.Conclusions:For the large area soft tissue defect wound of lower extremity, complicated with multiple deep tissues such as bone, joint and internal materials exposed, the greater omentum free transplantation under laparoscope combined with medium thick skin graft second stage has the advantages of good appearance and function after wound healing, less donor injury and fewer postoperative complications.

Objective To study the characteristics of hyperphenylalaninemia (HPA) and the differences in blood and urine metabolic index and their correlation.Methods A total of 137 patients with HPA diagnosed by the Pediatric Inherit Metabolism and Endocrine Department,Guangdong Women and Children's Hospital,Guangzhou Medical University from January 2014 to June 2017,were enrolled.Tandem mass spectrometry (MS/MS),gas chromatography/ mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC) were used to analyze the concentration of blood and urine metabolites in children,and the patients were divided into different groups according to the drug load test of tetrahydrobiopterin (BH4) and dihydrobiopterindine reductase (DHPR) deficiency.The HPA metabolite analysis of horizontal concentration by statistical differences and correlation analysis were performed.Results Among the 137 cases of HPA,there were 101 cases (73.7％) of phenylalanine hydroxylase deficiency (PAH),and among them 21 cases (15.3％) were classic phenylketonuria (PKU),37 cases were mild PKU (27.0％),43 cases (31.4％) wcrc mild HPA.Thcrc were 22 cases (16.1％) with BH4 reaction,and 79 cases (57.7％) of non-reactive type.Besides,there were 36 cases (26.3％) of tetrahydrobiopterin deficiency (BH4 D),of which 6-pyruvoyl tetrahydropterin synthase deficiency (PTPS) in 34 cases (24.8％) and dihydrobiopterindine reductase deficiency (DHPR) in 2 cases (1.5％).Urinary phenylacetic acid (r =0.673,P ＜ 0.01),phenyllactic acid (r =0.736,P ＜ 0.01),phenylpyruvic acid (r =0.642,P ＜ 0.01) were significantly correlated with blood phenylalanine (Phe) concentration,and the neopterin (N) (r =0.442,P ＜ 0.01) and biopterin (B) (r =0.398,P ＜ 0.01) had low correlation.Urinary phenylacetic acid,phenyllactic acid and phenylpyruvic acid had no correlation with urinary pterin.There were significant differences among PTPS deficiency group,BH4 response type,and non-reactive type(all P ＜ 0.05),but no significant difference between the BH4 reaction type and the non-reactive group (P ＞ 0.05).Conclusions Through the analysis of the different types of HPA metabolic profiles,it can help to master the incidence and characteristics in the region,within a certain concentration range of blood Phe,the phenylacetic acid,phenyllactic acid,phenylpyruvic acid should not be tested by GC-MS alone.Uterine erythropoietin analysis of BH4D classification and identification of BH4 reaction,non-reactive PKU have a supporting role,so master the metabolic index of various types of concentration and relevance of HPA,it can provide basis for early diagnosis,accurate treatment and follow-up.

Objective To investigate the class I integrons and their gene cassettes of imipenem-resistant Pseudomonas aeruginosa (IRPA) , and to analyze the correlation between integrons and drug resistance. Methods PCR was used to determine the presence of integrase genes and class I integrons. The variable regions were detected by sequencing. Resistance genes of integron gene cassettes including metal-β-lactamases, aminoglycoside modifying enzymes (AMEs), 16SrRNA methylating enzyme and the OprD2 genes were detected by PCR. The VITEK-2 automated system was used to determine the antibiotic susceptibility of integron-positive IRPA strains. Results The positive rates of integrase genes and class I integrons were 23.3%(20/86)and 8.14%(7/86) , and five kinds of gene cassettes were detected in 86 IRPA strains. The class I integrons-positive bacterial strains exhibited different resistant patterns to 12 antibiotics with large number of resistance genes. Conclusion The class I integrons and their gene cassettes are associated with multiple drug resistance of IRPA.

Objective To investigate clinical, imaging features, and long-term outcomes in patients with isolated anterior cerebral artery (ACA) territory infarction due to ACA atherosclerosis, and compare with isolated ACA territory infarction due to other etiologies. Methods The consecutive patients with acute isolated ACA territory infarction confirmed by diffusion-weighted imaging were enrolled prospectively. According to their stroke etiology, they were divided into ACA atherosclerotic stroke and non-ACA atherosclerotic stroke. The infarction patterns were classified as single infarction including perforating artery infarction (PAI), small branch infarction (SBI) and cortical branch infarction(CBI), and multiple infarctions (a combination of PAI,SBI or CBI).The clinical,imaging features and long-term outcomes were compared between the ACA atherosclerotic stroke group and the non-ACA atherosclerotic stroke group. Results A total of 86 patients (47 males) were enrolled, ages ranging from 39 to 88 years (mean 67.5 ± 12.5 years). There were 56 patients in the ACA atherosclerotic stroke group, and 30 patients in the non-ACA atherosclerotic stroke group (12 carotid atherosclerosis, 6 cardioembolism, 2 internal carotid artery dissection, 10 undetermined etiology). The proportions of females (53.6% vs. 30.0%; P= 0.043), progressive onset of stroke(58.9% vs. 20.0%;P=0.001),SBI alone(21.4% vs. 3.3%;P=0.029)and infarction involving small branches(80.4% vs. 46.7%;P=0.001)in the ACA atherosclerotic stroke group were higher than those in the non-ACA atherosclerotic stroke group, and CBI alone (17.9% vs. 55.3%, P=0.001) was lower. The follow-up times in the ACA atherosclerotic stroke group and the non-ACA atherosclerotic stroke group were 29.8 ± 16.5 months and 30.4 ± 18.5 months, respectively (P=0.534). Five-year cumulative incidence of adverse events (stroke, cardiovascular events and death) in the ACA atherosclerotic stroke group and the non-ACA atherosclerotic stroke group were 36.3% and 69.9% respectively(log rank test,P=0.021).Conclusions ACA atherosclerosis is the common etiology for isolated ACA territory infarction. The isolated ACA territory infarction due to ACA atherosclerosis had distinctive infarction patterns and a lower long-term incidence of adverse events compared with those due to non-ACA atherosclerosis.

Objective To investigate the risk and predictors of stroke recurrence in patients with symptomatic intracranial internal carotid artery(IICA)stenosis.Methods Consecutive patients with first-ever ischemic stroke or transient ischemic attack (TIA) caused by IICA atherosclerotic stenosis were enrolled prospectively. The patients were regularly followed up to assess stroke recurrence. Results A total of 70 patients were enrolled, 49 patients were males, and the mean age was 68.2 ± 12.3 years. The mean follow-up time was 34 ± 17 months (median, 33 months). Twenty-seven patients (38.6%) experienced recurrent events during the follow-up period (5 TIAs and 22 ischemic strokes);92.6% of recurrent events occurred in the original symptomatic stenotic IICA territory. Internal watershed infarction in patients with recurrent stroke was more common than those without stroke recurrence(74.1% vs. 44.2%,P=0.025). Kaplan-Meier survival analysis showed that the risks of stroke recurrence at 1,3 and 5 years were 26.8%, 42.5%, and 46.9%, respectively in patients with symptomatic IICA stenosis. Multivariate Cox proportional risk regression analysis showed that the predictors for stroke recurrence in patients with symptomatic IICA stenosis included diabetes (hazard risk [HR] 3.68,95% confidence interval[CI] 1.43-9.46; P=0.007), combined asymptomatic intracranial artery occlusive disease(HR 2.95,95% CI 1.16-7.50;P=0.023),and internal watershed infarction (HR 4.50, 95% CI 1.43-14.17; P=0.010) after adjusting for sex, age and traditional vascular risk factors. Conclusions The risk of long-term stroke recurrence in patients with symptomatic IICA stenosis is still high under the current drug treatment. Diabetes, combined asymptomatic intracranial arterial occlusive disease, and internal watershed infarction are closely associated with stroke recurrence.

Objective To investigate the differences of risk factors,imaging features,etiologies,and long-term outcomes in young adults with anterior and posterior circulation ischemic stroke.Methods Consecutive young patients (15-45 years) with ischemic stroke were divided into an anterior circulation group and a posterior circulation group.They were followed up regularly for a long term,and the endpoint events included stroke,cardiovascular events,and death.Results A total of 289 patients were enrolled in the study,including 220 males.Their mean age was 38.0 ±6.5 years.There were 213 patients (73.7％) in the anterior circulation group and 76 (26.3％) in the posterior circulation group.In terms of risk factors,valvular heart disease was more common in the anterior circulation group (6.6％ vs.0.0％;Fisher's exact test,P =0.025),while hypertension (51.3％ vs.36.6％;x2 =5.021,P=0.025) and prodromic infection (6.6％ vs.1.4％;Fisher's exact test,P =0.018) were more common in the posterior circulation group.In the etiologies of stroke,large-artery atherosclerosis was more common in the anterior circulation group (32.4％ vs.13.2％;x2 =10.435,P =0.001),while small vessel occlusion (26.3％ vs.15.5％;x2 =4.381,P =0.036) and arterial dissection (19.7％ vs.9.9％;x2 =5.012,P =0.025) were more common in the posterior circulation group.There was no significant difference in the incidence of 5-year cumulative end-point events between the anterior circulation group and the posterior circulation group (20.2％ vs.18.5％;log-rank test P =0.614).Multivariate Cox proportional hazards regression analysis showed that after adjusting for age and gender,the independent predictors of end-point events in the posterior circulatory group included hypercholesterolemia (hazard ratio [HR] 3.622,95％ confidence interval [CI] 1.216-17.766;P =0.030),old infarction (HR 6.045,95％ CI 1.602-29.580;P =0.016),and CE (HR 8.256,95％ CI 1.398-27.302;P=0.029).Conclusion There were significant differences in the risk factors,etiologies,and influencing factors of long-term outcome between the anterior circulation and posterior circulation in Chinese young patients with ischemic stroke,suggesting that the different diagnosis and treatment strategies should be adopted for the two types of patients.

Objective To investigate the effect of up-regulation of miR-132 on expressions of angiopoietin-1 (Ang-1)/endothelium-specific tyrosine kinase receptor 2 (Tie2) in focal cerebral ischemia reperfusion injury of rats.Methods Forty adult healthy SD rats were randomly divided into sham-operated group,cerebral ischemia group,miR-132 mimic group and negative control group (n=10).The models of middle cerebral artery occlusion in the later three groups were established by using modified Longa suture method.Rats in the miR-132 mimic group and negative control group were injected miR-132 mimic 15 μg and negative control 15 μg via paracele.Rats in each group were sacrificed 24 h after ischemia,and the brain tissues were collected;the total infarct volumes were calculated by TTC staining.The mRNA expressions ofmiR-132,Ang-1 and Tie2 in ischemic cerebral cortex tissues were detected by real-time fluorescence quantitative PCR.The protein expressions of Ang-1,Tie2,CD31 and vessel endothelial growth factor (VEGF) in ischemic cerebral cortex tissues were detected by Western blotting.Results The total infarct volume in the miR-132 mimic group was (27.92±3.05) mm3,which was significantly smaller than that in the cerebral ischemia group and negative control group ([51.34±2.86] mm3 and [50.46±2.57] mm3,P＜0.05).The relative mRNA expression levels of miR-132,A ng-1,Tie2,and the protein expression levels of Ang-1,Tie2,CD31,VEGF in the ischemic cerebral cortex tissues of the miR-132 mimic group were significantly higher than those in the negative control group,cerebral ischemia group and sham-operated group (P＜0.05);and those in the negative control group and cerebral ischemia group were significantly higher than those in the sham-operated group (P＜0.05).Conclusion Up-regulation ofmiR-132 expression could improve the ischemic states of ischemic stroke in rats,which might be related to Ang-1/Tie2 increased expressions to promote angiogenesis in ischemic brain tissues.

Objective To observe the curative effects of pterygium excision combined with autologous limbal stem cell transplantation in the treatment of primary pterygium.Methods 50 cases (72 eyes) of primary pterygium were selected in our hospital.According to the method of random number table,50 canses were divided into the control group (37 eyes) underwent pterygium excision and the study group (35 eyes) received autologous limbal stem cell transplantation on the basis of the control group,with 25 cases in each,and the efficacy were compare between the two groups.Results The healing time of study group[(3.02±0.37)d] was significantly shorter than in the control group[(6.85±0.48)d](t=2.95,P=0.01) and the recurrence rate (20%) was significantly lower than in the control group (52%) (x2=10.35,P=0.03),and the postoperative complications rate (8%) was significantly lower than in the control group (36%) (x2=13.94,P=0.03).Conclusion Compared with pure pterygium excision,pterygium excision combined with autologous limbal stem cell transplantation in the treatment of primary pterygium has higher clinical efficacy,which can effectively shorten the repair time and reduce recurrence rate,with high safety,so it is worthy of further promotion and application in clinic.

Objective · To prepare recombinant human TIGIT protein in E.coli and characterize its ability in binding Fusobacterium nucleatum (Fn). Methods · The gene of immunomodulatory protein of human TIGIT was amplified and cloned into pGEX4T2, and recombinant plasmid was transformed into E.coli BL21 (DE3) for GST-TIGIT fusion proteins were purified by the GST affinity chromatography and the interaction between GST-TIGIT fusion protein and Fn was tested by a pulldown assay. Results · Recombinant GST-TIGIT fusion protein expressed successfully in E.coli and was purified to homogeneity by GST affinity column. This protein could specifically bind to Fn, but not Lactobacillus acidophilus. Conclusion · High purify and activity of human GST-TIGIT fusion protein can be achieved by the prokaryotic expression system, and the adhesion between this protein and Fn has been preliminarily explored, which provides basis for further characterize interaction between them.