Objective To develop a rapid, sensitive and specific assay based on multiplex real-time PCR for detecting and identifying Escherichia coli O157: H7. Methods The lipopolysaccharide gene (rJbE) and H7 flagellar antigen gene(fliC) of Escherichia coli O157:H7 was chosen as targets, and then the primers and TaqMan-MGB probe were designed. The 5'end of probes was labeled with FAM and HEX fluo-resceins respectively; the 3'end of probes was labeled with MGB. The PCR reaction was optimized systemati-cally. Then the specificity, sensitivity and reproducibility of multiplex real-time PCR were estimated. Final-ly, multiplex real-time PCR was applied to detected clinical specimens. Results Escherichia coil O157:H7 were detected by multiplex real-time PCR accurately and quickly, which could distinguish Escherichia coli O157:H7 from O157: non-H7. Meanwhile, none of other bacteria could be identified. The sensitivity was 10 CFU/ml in pure culture. The coefficient of variation of intra-assay and inter-assay was less than 5%. When this assay was applied directly to identify 66 clinical specimens, the results showed that t5 were positive to Escherichia coil O157:H7 and 2 were positive to Escherichia coil O157: non-H7, in which 16 was the same to the results obtained from the conventional assays. The coincidence was 98.49%. Conclusion It is showed that multiplex real-time PCR is a reliable, accurate and feasible assay for detecting and identifying Escherich-ia coli Oi57: H7, The assay reported here provided a tool for analysis and diagnosis in the field of detecting clinical pathogens, epidemiologic survey and food safety monitoring.

Objective To explore the role of VEGF positive expression in tumor tissue in the prognosis of liver transplantation for hepatocellular carcinoma (HCC).Method Fifty cases of liver transplant recipients with HCC confirmed immunohistochemically were enrolled in this study.The MaxVisionTM two-step method was applied to detect the expression of vascular endothelial growth factor(VEGF),and the microvessel density (MVD) was measured in para-cancerous tissues by using DAB staining.The correlation of the VEGF tumor tissue in tumor tissue with Child-Pugh,MELD,tumor diameter and number,differentiation,MVD,Milan criteria and UCSF criteria for HCC liver transplantation was analyzed.Result In the HCC tissue,the VEGF positive expression rate was 52％(26/50).The one-year survival of recipients positive and negative for VEGF was 78％ and 100％,respectively,and one-year recurrence rate was 32％ and 12％,respectively,with the difference being significant (P =0.043 and P =0.048 respectively).The expression of VEGF was associated with Child-Pugh,tumor diameter,MVD,Milan criteria and UCSF criteria (P＜0.05 for all).Logistic regression analysis showed that low differentiation and VEGF positive expression were independent prognostic factors for HCC recurrence after liver transplantation.Conclusion VEGF has a certain reference value to judge HCC invasiveness and prognosis of liver transplantation.

Objective To evaluate the performance of virtual touch tissue quantification (VTQ)technique in noninvasive assessment of esophageal varices (EV) in patients with hepatitis B cirrhosis.Methods One hundred and thirty-five patients were enrolled in this study.Liver shear wave velocity (LSWV) and spleen shear wave velocity (SSWV) were obtained by VTQ technique,and the detection of EV was performed by upper endoscopy per patient.The severity of EV was staged on a G1-G3 scale.Liver function was also tested.ResultsThe mean value of LSWV in patients with cirrhosis was (2.48±0.50)m/s,LSWV was not correlated with the stage of EV.The mean value of SSWV was (3.25 ± 0.44) m/s,a significant,direct,linear correlation ( r =0.66,P＜0.001 ) was found between SSWV and the stage of EV.The cut-off value of SSWV for predicting the presence of EV was 3.16 m/s and the area under ROC curve was 0.83,with the sensitivity and specificity of 0.83 and 0.80,respectively.The cut-off value of SSWV for predicting severe EV was 3.39m/s and the area under ROC curve was 0.84,with the sensitivity and specificity of 0.83 and 0.77,respectively.ConclusionsIn cirrhotic patients infected with hepatitis B virus,SSWV measured by VTQ technique can be used as a noninvasive method for determining the presence and the severity of EV,however evidence to support a similar role for LSWV is lacking.

ObjectiveTo explore the application and significance of assisted laparoscopic hepatectomy (ALH) in living-donor-hepatectomy.MethodsWe successfully performed 7 cases of ALH of right hepatectomy on living donor from 30/5/2011 to 1/9/2011.ResultsThe donors recovered well with ratio of remnant lver:32.10％ ～38.31 ％,good liver fuction,little pain and no surgical complications.All the wound sutured intracuteneously was taken out stitches 7 days after operation and healed perfectly.Liver function got normal 2 weeks after operation.Conclusions ALH,which gives the consideration to both the minimal invasion of laparoscopic surgery and safe of open surgery,can be applied safely in hepatectomy of living donor and highly acceptible for donor and receptor.

OBJECTIVEBenign epilepsy with centro-temporal spikes (BECTs) is a common idiopathic partial epileptic syndrome in childhood, which often affect the pre-school and school-age children and a considerable proportion have comorbidity including lower academic achievement and cognitive impairment. Few studies involved the psychocognitive assessment in such a drug-treatable epileptic syndrome especially in the newly diagnosed and medications-naive group. This study aimed to investigate the cognitive characteristics of children with newly onset BECTs before treatment.

METHODForty-one outpatients with newly diagnosed BECTs who visited the Clinic during the periods from October 2012 to May 2014 before the medications against epilepsy and 41 healthy controls recruited from regular school in Beijing during the period from July 2013 to March 2014, who matched in age and gender underwent battery testing by computerized cognitive testing in epilepsy (CCTE). The BECTs group included 41 children, 20 boys and 21 girls, mean age (8.2 ± 1.7) years, the age of onset of epilepsy 4.5-11.5 years (the age of onset <8 years in 25 cases, ≥ 8 years in 16 cases). The cognitive characteristics and associated factors were analyzed. The primary data including correct answer numbers and reaction times were analyzed by independent sample t-test between the two groups of children with BECTs and healthy controls based on SPSS 18.0 statistical software.

RESULTRaw data of 9 tasks' scores collected from BECTs and healthy control children were continuous variables in accordance with normal distribution. BECTs children performed significantly worse than controls in choice reaction time ((618+158) vs. (524+254) ms), three-dimensional mental rotation (11 ± 10 vs. 18 ± 12) and visual tracing (10 ± 6 vs.15 ± 6), t=2.01, 3.03 and 3.47, P<0.05, <0.01 and <0.001, respectively.While other 6 tasks showed no significant difference between the two groups (P>0.05 for all comparisons). BECTs boys performed significantly worse than girls on simple substraction tasks compared with standard nine score ((4.7 ± 1.5) vs. (5.6 ± 1.2), t=-2.24, P<0.05). Other 8 tasks showed no significant difference between boys and girls (P>0.05 for all comparisons). Other 9 tasks showed no significant differences between the two groups of BECTs children whose age of onset was before 8 years and those who started seizure ≥ 8 years (P all >0.05). The standard nine scores of simple substraction from the three BECTs groups of dominance sides of spikes and waves during NREM showed significant difference (P<0.05). BECTs children with bilateral discharges performed significantly worse than the other two groups dominantly right or left discharges (4.7 ± 1.2 vs. 6.0 ± 1.2 vs. 4.9 ± 1.4, P all <0.05). There was no significant difference between the two groups with right and left side dominance discharges (P>0.05). Other 8 tasks showed no significant differences among the three groups (P>0.05 for all comparisons).

CONCLUSIONAlthough EEG discharges index below 50% during NREM period, while newly diagnosed BECTs children before treatment with medications against epilepsy performed poorer on tasks of choice reaction time, three-dimensional mental rotation, and visual tracing. The two factors of male and bilateral discharges during NREM period correlate with dysfunction of simple subtraction, the mechanism needs further study and the cognitive function of epilepsy children should be evaluated and followed up, in order to provide psychologic baseline data for persistent cognitive disturbance.

Beijing ; Case-Control Studies ; Child ; Child, Preschool ; Cognition ; Cognition Disorders ; diagnosis ; Comorbidity ; Epilepsy, Rolandic ; physiopathology ; Female ; Humans ; Male ; Reaction Time ; Seizures ; physiopathology

Objective To evaluate the therapeutic effect of computerized online cognitive training for children with attention deficit hyperactivity disorder (ADHD).Methods Fifteen children with ADHD recruited from Department of Neurology in the Children's Hospital Affiliated to Capital Institute of Pediatrics received computerized online cognitive training including attention training,visual spatial training and memory training.Cognitive neuropsychological test battery (choice reaction time,mental rotation,word semantic,simple subtraction,working memory and visual tracing) was used to assess the cognitive function in pre-training and post-training stages,such as basic response ability,reaction speed,visuo-spatial cognitive ability,semantic comprehension,calculation fluency,working memory and attention ability.Paired-samples t test was used to make comparison of the cognitive test results between pre-training condition and post-training condition.Results All the correction scores of cognitive tests were transformed to percentile scores.Paired-samples t test results showed that the ADHD children got higher scores in the post-training condition than in the pre-training condition in working memory test [(52.00 ± 20.77) ％ vs (39.73 ± 23.64) ％,t =2.72,P ＜ 0.05].There was no significant training effect in choice reaction time,mental rotation,sentence completion,simple subtraction and visual tracing(all P ＞ 0.05).Conclusions Computerized online cognitive training can significantly improve the working memory of children with ADHD,and has no side effects on other cognitive functions.It need to further expand the sample size and follow up the training effect for a long-term.

Objective To discuss the modified eversion thrombectomy for portal vein thrombosis (PVT) in liver transplantation and the curative effectiveness.Methods All 613 cases complicated with PVT preoperation were given modified eversion thrombectomy,and there were 179,236,182 and 16 cases of PVT Yerdel grade Ⅰ,Ⅱ,Ⅲ and Ⅳ respectively.Results All 415 PVT patients of grade Ⅰ and Ⅱ received modified eversion thrombectomy and success rate was 100％.Among 182 PVT patients of grade Ⅲ received modified eversion thrombectomy (success in 176 cases,and failure in 6 cases).Sixteen PVT patients of grade Ⅳ received modified eversion thrombectomy (success in 13 cases and failure in 3 cases).The 1-year survival rate of PVT patients after surgical operation was 93.6％,and that of non-PVT patients was 94.6％ (P＞0.05).Conclusion Modified eversion thrombectomy could be adopted in PVT patients regarding to different Yerdel grades.

Objective To compare the effect of mechanical perfusion and simple cold preservation on donation after cardiac death (DCD)pancreas in pigs.Methods Ten healthy pigs were randomized into simple cold preservation group and the mechanical perfusion group (5 pigs in each group ).DCD model was established in pigs.The pancreas was cut and stored in University of Wisconsin solution (UW solution).The pancreas of the simple cold preservation group was preserved with simple UW solution and that of the mechanical perfusion group was preserved by mechanical perfusion.Specimen was collected from pancreatic tail at 1,2,3,4,6 and 24 h to prepare tissue section.Then,the tissue section was stained with hematoxylin-eosin (HE).Histopathological examination was conducted and pathological score of the two groups was compared.Results Microthrombus in DCD pancreas of pig was removed at 180 min of mechanical perfusion and injury to pancreas islet caused by excessive perfusion was avoided.The pathological score of the mechanical perfusion group was (4.2 ±0.8)and that of the simple cold preservation group was (8.4 ±1.1),and the difference had statistical significance (P <0.05).Conclusions Mechanical perfusion may effectively remove thrombus in pancreatic vessels.Compared with simple cold preservation,mechanical perfusion may maintain the integrity of pancreas islet better after the preservation of the same period of time.

Objective     To compare the short- and long-term survival of patients with stage T1N0M0 non-small cell lung cancer (NSCLC) undergoing robot-assisted thoracic surgery (RATS) and video-assisted thoracoscopic surgery (VATS). Methods     The clinical data of 396 patients with stage T1N0M0 NSCLC treated with RATS or VATS in our hospital from 2012 to 2019 were retrospectively analyzed. There were 209 males and 187 females, with a mean age of 61.58±8.67 years. According to surgical procedures, they were separated into two groups: a RATS group (n=157) and a VATS group (n=239). The two groups were compared in terms of the survival and prognosis-influencing factors. Results     The intraoperative blood loss and postoperative 24 h drainage volume in the RATS group were less than those  in the VATS group (48±42 mL vs. 182±231 mL, P<0.001; 250±119 mL vs. 324±208 mL, P<0.001). The groups and number of dissected lymph node in the RATS group were more than those of the VATS group (5±2 groups vs. 3±2 groups, P<0.001; 17±9 vs. 11±8, P<0.001). There was no statistical difference in the postoperative 48 h drainage volume (P=0.497), postoperative intubation time (P=0.180) or hospital stay (P=0.313). The survival state and recurrence-free survival state in the VATS group were better than those in the VATS group (1-year survival rate: 98.7% vs. 94.8%, 5-year survival rate: 90.5% vs. 75.8%, 8-year survival rate: 76.9% vs. 62.1%, mean survival time: 93 months vs. 79 months, P=0.005; 1-year recurrence-free survival rate: 97.4% vs. 95.6%, 5-year recurrence-free survival rate: 94.8% vs. 77.8%, 8-year recurrence-free survival rate: 82.6% vs. 64.8%, mean recurrence-free survival time: 95 months vs. 79 months, P=0.004). Univariate analysis showed that surgical method, the groups and the number of dissected lymph nodes were the influencing factors for postoperative overall survival and recurrence-free survival. At the same time, the results of multivariate analysis showed that surgical method was a common independent factor for overall survival and recurrence-free survival. Conclusion     RATS can obtain better survival in patients with T1N0M0 NSCLC, and RATS has more thorough lymph node dissection, less intraoperative blood loss and postoperative 24 h drainage volume.

Objective: To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks. Methods: Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman’s rank correlation coefficient was used to test bivariate associations. Results: Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29). Conclusion Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.