This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.

Background:Acute gastroenteritis is the second largest public health problem in the world. Rotavirus(RV)is one of the pathogens of acute gastroenteritis in adults,researches focusing on RV infection may provide the basis for prevention and control of the disease. Aims:To determine the epidemiological characteristics of RV infection in adults with acute gastroenteritis in Shanghai,China. Methods:From Jun. 2010 to Dec. 2013,stool samples and clinical data in adults with acute gastroenteritis in a designated hospital in Shanghai Changning District were collected. ELISA and PCR were used to detect RV infection and its genotypes. Results:A total of 1 554 eligible stool samples from acute gastroenteritis patients were recruited,of them 691 were males and 863 were females,the mean age was(46. 19 ± 15. 59)years old. RV was detected in 189 patients with a detection rate of 12. 2% ,163(10. 5% )were categorized as group A RV and 26(1. 7% ) were group B/ C RV;the most common genotypes in group A RV were G9(30. 1% )and G1(25. 2% ). Watery stool and vomiting were more prevalent in RV-positive patients than in RV-negative patients(P < 0. 05). The detection rates in years 2010,2011,2012 and 2013 were 12. 2% ,14. 9% ,6. 8% and 16. 3% ,respectively. When analyzed by age group,the detection rate was significantly lower in 18-39 years group than those in 40-59,60-79,and ≥80 years groups(8. 7% vs. 14. 8% ,14. 2% ,and 17. 1% ,P < 0. 05). The peak of epidemic was from Nov. to next Feb. Conclusions:RV infection in adults with acute gastroenteritis is more popular in middle aged and elderly people and shows a winter seasonality in Shanghai,China. The most common genotypes of group A RV are G9 and G1.

Objective To investigate the effect of remote ischemic postconditioning (RIPostC ) on the platelet reactivity in the patients with acute myocardial infarction (AMI) receiving reperfusion therapy .Methods Seventy-one cases of AMI entering the group received the reperfusion therapy .The patients were divided into the two groups .The treatment group received RIPostC ,while the control group received sham RIPostC .The venous blood samples were collected before transcutaneous coronary intervention (PCI) ,instantly after RIPostC and at 24 h ,48 h after PCI .The platelet activation indicators CD62P and PAC-1 ,and platelet apopto-sis indicator mitochondrial transmembrane potential(ΔΨm) were measured by flow cytometry .Results CD62P expression on plate-let surface at 24 h after PCI in the treatment group was significantly lower than that in the control group (P<0 .05) ,but which at other time points had no statistical difference between the two groups (P>0 .05);there was no statistical difference in platelet PAC-1 expression at each time point between the two groups (P>0 .05);the platelet ΔΨm at each time point had no statistical difference between the two groups(P>0 .05) .Conclusion RIPostC can somewhat reduce platelet activation in AMI patients without causing platelet early apoptosis .

Objective To evaluate the value of stroke prognostication using age and National Institute of Health Stroke Scale index (SPAN) for outcome after early endovascular treatment for anterior circulation large vessel occlusion.Methods The patients who underwent early endovascular treatment were prospectively,sequentially collected in Yijishan Hospital of Wannan Medical College from December 2014 to September 2017 and Jinling Hospital from March 2014 to March 2017.Individuals whose age in years plus NIHSS score was greater than or equal to 100 were designated as SPAN-100-positive patients,while those with a score less than 100 were designated as SPAN-100-negative patients.We compared the baseline data and perioperative data between the two groups.The 90 days modified Rankin Scale score≤2 was regarded as favorable outcome.Single factor and multivariable Logistic regression analyses were used to determine the association between SPAN-100 and outcomes.Results One hundred and ninety patients were enrolled,20 (10.5％) of which were SPAN-100 positive,and 170(89.5％) were SPAN-100 negative.There were no significant differences between the two groups on postoperative intracerebral hemorrhage and 90 days mortality.Ninety days independence rates were higher in SPAN-100-negative patients (77/170,45.3％) than in SPAN-100 positive patients (4/20,20.0％;x2 =4.681,P =0.030).Multi-factor Logistic regression analysis showed that the higher preoperation systolic pressure (OR =1.030,95％ CI 1.008-1.052,P =0.007),the lower Alberta Stroke Program Early CT Score (OR =1.609,95％ CI 1.056-2.453,P =0.027) and poor collateral circulation(OR =5.714,95％ CI 1.668-19.570,P =0.006) were the independent risk factors of outcomes.Conclusion SPAN-100 is not an independent predictor of favorable outcome after adjusting for factors of outcomes in patients with anterior circulation large vessel occlusion.

Objective To evaluate the level of daily red meat intake and prevalence of excessive red meat intake among employees of floating population in China.Methods 48 511 employees of floating population aged 18 to 59 from 170 counties of 31 provinces (autonomous regions and municipalities) and Xinjiang Production and Construction Corps (District) were selected by stratified cluster sampling method.Information on red meat intake was collected by semiquantitative food frequency questionnaire.Average intake of 100 g/day,recommended by the World Cancer Research Fund,was used as the cut-off point to estimate the prevalence of excessive red meat intake.After performing the complex weighted analysis,level of daily red meat intake and prevalence of excessive red meat intake were calculated by demographic characteristics including age,education,industries and body mass index etc.Results 1) The mean daily red meat intake was 125.9 g(95％CI:116.5 g-132.5 g),higher in men(141.6 g,95％CI:131.3 g-148.9 g) than in women (104.7 g,95％CI:95.8 g-111.2 g) (P＜0.01).Results from the Tendency Test did not show statistically significant changes on the red meat intake related to age,education level or body mass index (P values for trend were all greater than 0.05).The standardized mean daily intake of red meat,adjusted by 2010 census data of China,was 121.0 g(95％ CI:113.4 g-128.7 g).2) The prevalence of excessive red meat intake was 36.2％ (95％ CI:33.0％-39.3％) significantly higher in males (42.4％,95％ CI:38.9％-45.8％) than in females (27.8％,95％CI:27.1％-31.0％) (P＜0.01).The prevalence was estimated to be the highest among the population aged 30-39,with 43.5％ (95％CI:39.7％-47.4％) in males and 30.1％(95％CI:26.5％-33.9％) in females.The standardized prevalence,adjusted by 2010 census data of China,appeared to be 34.6％ (95％CI:31.9％-38.0％).Conclusion The level of daily red meat intake was higher than 1 00 g/d,the standard recommended by the World Cancer Foundation,among floating population of China.Both the mean daily red meat intake and prevalence of excessive red meat intake were higher in floating population than that in the local residents in China.

In this paper, the current situation and existing problems of spinal surgery practice are analyzed and summarized. Combined with the review of the structure and function of spinal region, we have expounded the application and advantages of Essential Anatomy software in the clinical practice teaching of spinal surgery. This software has a friendly interface, simple operation, real content, intuitive and visual model, and it can provide videos and slice function, which makes the original complex and abstract spinal anatomy practice content become novel and vivid, and greatly deepens doctors' understanding of the structure, function, movement and the disease pathogenesis of spinal region. At the same time, it also greatly mobilizes the learning interest and initiative of the clinical practice doctors, and improves the effect of clinical practice.

Objective:To investigate the relationship between the down-regulation expression of sodium taurocholate cotransporting polypeptide (NTCP) in proliferating hepatocytes and the response to antiviral therapy of chronic hepatitis B (CHB) patients.Methods:Sixty-eight hepatitis B e antigen (HBeAg)-positive CHB patients admitted to the Fifth Medical Center of PLA General Hospital from January 2011 to March 2015 were included. Basic information and laboratory data were collected. Based on the baseline (before antiviral treatment) inflammatory activity (G), the patients were divided into ≤G2 group and >G2 group. Twelve liver puncture tissue samples were selected from each group for NTCP and Ki67 immunofluorescence staining.The proportion of Ki67-positive cells was calculated, and the staining of NTCP was scored. Five pairs of tissue specimens of patients who had hepatitis B virus (HBV) infection were diagnosed with focal nodular hyperplasia (FNH) and underwent nodular resection surgery from March 2014 to March 2017 were collected.Immunohistochemical staining was used to detect the expressions of Ki67, NTCP and hepatitis B surface antigen (HBsAg) in each tissue specimen, and the proportion of staining positive cells or the staining intensity was calculated. Statistical analysis was performed by Mann-Whitney U test, chi-square test or Spearman correlation analysis. Results:The proportion of Ki67-positive cells (6.75%(6.20%, 8.16%))in five liver FNH tissues with HBV infection was significantly higher than that in adjacent non-FNH tissues (0.75%(0.66%, 1.20%)), while the immunohistochemical scores of NTCP and HBsAg (3.00 (1.00, 3.00) and 2.00 (1.00, 2.00), respectively) were both significantly lower than those in adjacent non-FNH tissues (8.00 (8.00, 9.00) and 8.00 (6.00, 8.00), respectively), the differences were all statistically significant ( Z=-2.611, -2.424 and -2.635, respectively, P=0.009, 0.015 and 0.008, respectively). There were 37 patients in >G2 group, and 31 patients in ≤G2 group. After six months of antiviral treatment, CHB patients with persistent inflammation in >G2 group obtained a better virological response, with serum HBV DNA and HBeAg showing a greater decline ((0.71±0.14) lg IU/mL and (0.92±0.13) lg IU/mL, respectively) than those in ≤G2 group ((0.54±0.30) lg IU/mL and (0.49±0.65) lg IU/mL, respectively) ( Z=-3.048 and -2.666, respectively, P=0.002 and 0.008, respectively). The proportion of Ki67-positive cells in the specimens of >G2 group (4.34%(1.84%, 8.77%)) was significantly higher than that of ≤G2 group (0.34%(0, 0.80%)) ( Z=-3.640, P<0.01), and the immunohistochemical staining score of NTCP (1.00 (0, 3.25)) was significantly lower than that of ≤G2 group (6.00 (4.00, 8.00)) ( Z=-3.012, P=0.003). Spearman correlation analysis showed that the NTCP immunohistochemical score was negatively correlated with the proportion of Ki67-positive cells ( r=-0.512, P=0.01). Conclusions:CHB patients with persistent inflammationare often accompanied by more active hepatocyte proliferation and low membrane NTCP expression, which is not conducive to HBV reinfection. It may facilitate these patients to obtain better virological response.

Objective:To investigate the early predictive factors of periprocedural thrombus migration and the relationship between periprocedural thrombus migration and prognosis after mechanical thrombectomy (MT) in stroke patients.Methods:The patients with anterior circulation acute large vessel occlusion stroke (ALVOS) who underwent MT in the Stroke Center of Yijishan Hospital of Wannan Medical College from May 2015 to December 2019 were retrospectively analyzed. The baseline characteristics, procedural and clinical outcomes were collected. Univariate and multivariate regression analysis was used to explore the risk factors of thrombus migration and the relationship between thrombus migration and prognosis of patients.Results:There were 302 ALVOS patients [(68.8±11.0) years old and 166 males (55.0%)] included, of whom thrombus migration was identified in 80 patients (26.5%), including 60 cases (75.0%) of proximal migration. Cardiogenic stroke ( OR=2.722, 95% CI 1.367-5.418, P=0.004) and clot burden score (CBS; OR=0.849, 95% CI 0.745-0.968, P=0.015) were independent risk factors of thrombus migration. Proximal migration ( OR=2.822, 95% CI 1.220-6.528, P=0.015) was an independent risk factor of 90-day clinical outcome, while the effect of distal migration on 90-day clinical outcome was not statistically significant. Conclusions:Cardiogenic stroke and lower CBS score are independent predictors of periprocedural thrombus migration in ALVOS patients who underwent MT. Proximal migration is an independent risk factor for the prognosis of patients, which has important clinical intervention significance.

ObjectiveTo investigate the serological markers associated with posthepatectomy recurrence in patients with hepatocellular carcinoma, and to establish a prognostic model to evaluate whether palliative hepatectomy is suitable for such patients. MethodsA total of 111 patients with hepatocellular carcinoma who underwent hepatectomy in the Affiliated Cancer Hospital of Zhengzhou University from February 2009 to July 2013 and received follow-up were enrolled. Basic clinical data were collected and the patients were divided into recurrence group and non-recurrence group according to whether recurrence was observed during follow-up. The t-test was used for comparison of normally distributed continuous data between two groups and the Wilcoxon rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. Survival curves were plotted using the Kaplan-Meier method, and survival differences were analyzed using the log-rank test. A Cox regression analysis was used to perform univariate and multivariate analyses, and the area under the ROC curve (AUC) was used to evaluate prediction efficiency. ResultsThe Kaplan-Meier survival curves showed that the patients with low alpha-fetoprotein (AFP), alkaline phosphatase, gamma-glutamyl transpeptidase (GGT), and fibrinogen and high CXCL13 had a longer median time to recurrence (P＜0.05). AFP (hazard ratio ［HR］［95%CI］=1.69(1.03~2.79), P=0.039), GGT (HR［95%CI］=1.89(1.14~3.14), P=0.014), and CXCL13 (HR［95%CI］=0.54(0.33~0.89), P=0.015) were independent factors associated with posthepatectomy recurrence. The prognostic index PI=0.526×AFP+0.637×GGT-0.616×CXCL13 established based on these factors had an AUC of 0.87, a sensitivity of 93.75%, and a specificity of 63.64% in predicting recurrence within 0-3 months after palliative hepatectomy, with a significant reduction in prediction efficiency for recurrence within 0-6 months (AUC=0.68) or a longer period of time. The recurrence prediction efficiency of this model for palliative hepatectomy was significantly higher than that for radical resection. ConclusionThe prognostic model established based on CXCL13, AFP, and GGT can be used to evaluate the risk of early recurrence after palliative hepatectomy and thus helps clinicians to make diagnosis and treatment decisions based on patients’ benefits.

Objective:To observe the incidence, risk factors of malignant brain edema (MBE) and the influence of MBE on outcomes after early successful recanalization of acute large vascular occlusion stroke (ALVOS).Methods:A total of 149 patients (age (68±11) years, male 85 (57.0%)) with ALVOS who underwent early endovascular treatment and achieved successful recanalization at the First Affiliated Hospital of Wannan Medical College from July 2014 to February 2019 were retrospectively analyzed. Baseline data, perioperative data, and 90-day prognostic information were collected from patients enrolled in the study. Univariate and multivariate analyses were used to explore the relationship between MBE and outcomes, and the risk factors of MBE.Results:Among the 149 patients, baseline National Institutes of Health Stroke Scale score was 16 (13, 20), baseline Alberta Stroke Project early CT score was 9 (8, 10), the time of onset-to-puncture was (248.3±61.3) minutes, and the onset-to-recanalization time was (312.4±69.7) minutes. MBE occurred in 23 patients (15.4%, 23/149). The 90-day favorable outcome (90-day modified Rankin Scale score≤ 2) in patients with MBE was significantly lower than those without MBE (17.4% (4/23) vs 61.1% (77/126), χ 2=14.985, P<0.001), and the 90-day mortality in patients with MBE was significantly higher than those without MBE (43.5% (10/23) vs14.3% (18/126), χ 2=10.861, P=0.003). MBE was shown to be an independent predictor of 90-day poor outcome (adjusted OR=12.078, 95 %CI 1.934-75.443, P=0.008) and death (adjusted OR=4.146, 95 %CI 1.060-16.216, P=0.041). Multivariate Logistic regression analysis showed that the collateral circulation status was related to the incidence of MBE in patients with ALVOS after successful recanalization (level 2 vs level 0, adjusted OR=0.109, 95 %CI 0.021-0.563, P=0.008). Conclusions:MBE is an independent risk factor of ALVOS patients with poor outcome or death in 90 days. For patients with ALVOS, even if the occlusive vessels have been successfully recanalized after early endovascular treatment, MBE is still not uncommon. The collateral circulation state is an independent predictive factor of the development of MBE after recanalization by early endovascular treatment in patients with ALVOS.