AIM: To establish a quantitative measurement method of ECV-304 cell migration in a scratch wound model in vitro. METHODS: The method of ECV-304 cell scratch wound model was improved. The ECV-304 cell migration was measured quantitatively using a computer-assisted video microscopic method with image-analysis system. The effects of heparin and thrombospondin-1 on ECV-304 cell migration were observed in the scratch wound model in vitro. RESULTS: The average width and length of cell scratch wound was (0.95?0.08) mm and (51.20?3.40) mm respectively in 120 experiments. The peak of cell migration reached about 24 h after culture. The stimulated effect of heparin and inhibited effect of thrombospondin-1(TSP-1) on cell migration were observed. CONCLUSION: A quantitative measurement method of ECV-304 cell migration in vitro is established. The stimulated and inhibited effects on ECV-304 cell migration were proven respectively using this method.

Objective To investigate the feasibility of using leukocytes that were filtered out by LeukoReduction System ( LRS) to replace conventional human peripheral blood leukocytes in experimental researches and to comparatively analyze the differences between them in vitro biological functions and pheno-types of T cells. Methods Mononuclear cells were isolated from LRS-separated leukocytes and whole blood sample that collected from the same person by using Ficoll. Fluorescence-activated cell sorting ( FACS) was performed to analyze the phenotypes of T cells. CD3+T cells were sorted out by using magnetic beads. The T cells that were collected by using two different ways were incubated with anti-CD3 and anti-CD28 antibodies and IL-2 in vitro for 10 days. Several assays including cell counting, FACS and cytometric beads array ( CBA) were performed to comparatively analyze the differences in biological functions and phenotypes of T cells that were isolated by different methods. Results The phenotypes of T cells isolated from LRS filter and whole blood sample were highly similar at the initial stage. The sorting rate of CD3+T cells form LRS filter reached a high level and met the requirements for experimental researches. No statistically significant differ-ences in cell count, phenotype, expression of costimulatory molecules and cytokine secretion were observed between T cells isolated from LRS filter and whole blood sample. Conclusion This study suggested that the T cells isolated from LRS filter could be used as an alternative to whole blood T cells for fundamental resear-ches since they were similar in cell vitality, phenotype and biological functions. It provided a new way to solve the problem of blood shortage in clinic and scientific research.

Objective To evaluate the outcome of patients with alcoholic liver disease (ALD) after orthotopic liver transplantation (OLT) and to study the prognostic factors.Methods The data of 17 patients who underwent OLT for ALD from January 2010 to March 2013 were analyzed retrospectively.The data on age,gender,history of gastrointestinal bleeding,history of splenectomy,Child-Pugh score,Maddrey' s discriminant function and MELD score were evaluated using the Kaplan-Meier method for univariate analysis.The log-rank test was applied to compare the survival rates.Results The overall survival rate at 100 weeks in patients less than 55 years old was 90％ (9/10),while that in patients more than 55 years old was 28.57％ (2/7).There was a significant difference between the two groups (P ＜ 0.05).There were no statistically significant differences between female and male patients,between patients with or without a history of gastrointestinal bleeding or splenectomy (P ＞ 0.05).There was no significant difference on survival rates after liver transplantation between patients with Child-Pugh B and Child-Pugh C,patients with Maddrey' s scores ＜ 70 and ≥70,and patients with a MELD score ＜ 30 and ≥ 30 (P ＞ 0.05).Conclusions Better survival rates were observed in ALD patients who were ＜ 55 years old after liver transplantation.The Child-Pugh score,Maddrey' s discriminant function and MELD score were of no prognostic significance.These findings still need to be verified by prospective large-scale studies.

Objective To establish a stable and reliable mouse model as an alternative to the traditional model of impaired glucose tolerance induced by calorie restriction and its effect on glucose homeostasis.Methods Forty 16-week-old SPF C57BL/6J mice (half male and half female) were randomly divided into four groups by sex and the way of feeding.The mice in the ad libitum (AL) group had free access to basic diet, while the mice in the intermittent fasting (IF) group had normal diet and fasting on alternate days, with free access to water on the fasting days.The changes of body weight and blood glucose concentration in each group were monitored, and intraperitoneal glucose tolerance test and insulin tolerance test in mice were performed before and after the 12-week IF treatment.Results At 12 weeks after IF treatment, the body weight and blood glucose concentration of mice did not show significant difference.After i.p.injection of glucose, the blood glucose concentration of IF mice was less increased than the AL group, and after the insulin injection, the blood glucose concentration was more decreased.Compared to the AL group, the areas under the curve of tolerance test in the IF group were significantly decreased (P < 0.05).Conclusions After IF treatment, the mice show an enhanced sensitivity to insulin and improved glucose tolerance.This establishment method of mouse model of intermittent fasting is easy and simple, therefore, can be used as an effective alternative to traditional calorie restriction model of impaired glucose tolerance.

Morels (Morchella spp.) are valuable medicinal and edible mushrooms. In this study, chemical profiles of aromas and lipophilic extracts of black morel (Morchella importuna) grown in China were analyzed by gas chromatography/mass spectroscopy, along with the evaluation of antioxidant and antimicrobial activities for the lipophilic extracts. Sixty-five compounds in total were identified from the aromas, and 1-octen-3-ol was the main component for aromas of fresh (34.40%) and freeze-dried (68.61%) black morels, while the most abundant compound for the aroma of the oven-dried sample was 2(5H)-furanone (13.95%). From the lipophilic extracts, 29 compounds were identified with linoleic acid as the main compound for fresh (77.37%) and freeze-dried (56.46%) black morels and steroids (92.41%) as the main constituent for an oven-dried sample. All three lipophilic extracts showed moderate antioxidant activities against 2,2-diphenyl-1-picrylhydrazyl and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) with the IC50 values ranging 7.56 ∼ 17.52 mg/mL and 5.75 ∼ 9.73 mg/mL, respectively, and no obvious antimicrobial activity was observed for lipophilic extracts. The drying methods affect the chemical profile of black morel, and freeze-drying was favorable for retaining nutrients and morel smell. This is the first report on the aroma and lipophilic extracts of M. importuna grown in China.

Objective: To compare the clinical effects between laparoscopic cholecystectomy and choledochotomy versus traditional open cholecystectomy plus choledochotomy. Methods: One hundred and sixty-eight elderly patients with gallbladder and choledocholithiasis were divided into a laparoscopy group(n=75, receiving laparoscopic cholecystectomy and choledochotomy)and an open abdominal group(n=93, undergoing traditional open cholecystectomy and common bile duct exploration). The surgical incision length, operation time, intraoperative blood loss, anal exhaust time, hospital stay and postoperative complications were compared between the two groups. Results: The surgical incision length, operation time, intraoperative blood loss, anal exhaust time, hospital stay were lower in the laparoscopic group than in the open abdominal group(P<0.05). The incidence of postoperative complications was lower in the laparoscopic group than in the open group(P<0.05). Conclusions Laparoscopic cholecystectomy and choledochotomy can obviously improve the clinical efficacy and reduce complications in elderly patients with gallbladder and choledocholithiasis, and it is worthy of clinical application.

Objective To investigate the clinicopathological and prognostic significance of lysosome-associated membrane protein-2a (LAMP2a) expression in the development of colorectal cancer.Methods Immunohistochemistry was performed to detect the LAMP2a expression in colorectal cancer,normal mucosa,and adenoma tissues.Results LAMP2a expression in colorectal cancer and adenoma was higher than that in normal mucosa (P ＜ 0.05).Further,LAMP2a expression in colorectal cancer was negatively correlated with TNM staging and lymph node metastasis (P ＜ 0.05);however,the invasion and liver metastasis,depth of invasion and differentiation had no correlation with the lymph node (P ＞ 0.05).Kaplan-Meier analysis showed that LAMP2a expression in colorectal cancer is not significantly correlated with good prognosis (P ＞ 0.05).Univariate survival analysis showed that lymphatic invasion,vascular invasion,lymph node metastasis,TNM stage,liver metastasis,distant metastasis,and differentiation were correlated with poor prognosis of colorectal cancer (P ＜ 0.05).The Cox proportional hazards regression model showed that the depth of invasion and distant metastasis were important factors affecting the survival period of patients with colorectal cancer (P ＜ 0.05).Conclusion LAMP2a may play a role in the carcinogenesis and progression of colorectal cancer,and may be used as a molecular marker for the biological behavior of colorectal cancer.

Objective: To understand effects of timing and duration of separation experiences from parents on emotion regulation of left-behind preschoolers,and to provide the reference for phychological instruction and intervention among the special groups of children. Methods: Children’s emotion regulation strategy and the relevant information among 1 373 left-behind preschool children from Funan county in Fuyang.Qianshan county in Anqing,Changfeng county and Fexi county inFeixi were investigated. Results: Children with left-behind experiences younger than 18 months old tend to use less cognitive restructuring (P=0.03) and alternative action strategies (P=0.00) than non-left behind children. Children separated from father less than 47 months (median) and 36 months (median) from mother tend to use less cognitive restructuring (P=0.00) and alternative action strategies (P=0.00) than non-left behind children. Conclusion Separation experiences from parents younger than 18 months old exert severe damage on children’s emotional regulation. With the duration of separation increases, children show resilience of emotion regulation, which might be a protective factor for negative emotion due to parent-child separation.

Objective:To explore the optimization strategy of the Asia-Pacific colorectal screening (APCS) scoring system in the screening of colorectal neoplasms.Methods:From February to Decomber in 2016 and March to December in 2018, at Xijing Hospital of Air Force Military Medical University and the First Affiliated Hospital of Xi′an Medical University, patients who received opportunistic screening colonoscopy were enrolled. Before colonoscopy, the APCS score (low-risk zero to one points, medium-risk two to three points and high-risk four to seven points), body mass index (BMI), fecal occult blood test (FOBT) and plasma methylated Septin9 gene ( mSEPT9) of all patients were detected and recorded. The results of colonoscopy and biopsy pathology were taken as the gold standard, the efficacies of the above methods in screening colorectal neoplasms were compared to determine and optimize the screening efficiency of APCS scoring system. Chi-square test was used for statistical analysis. Results:A total of 494 patients were screened, of whom 133 cases were diagnosed with colorectal polyps, including 86 cases of colorectal adenomatous polyps (82 cases of non-progressive adenoma, and four cases of advanced-adenoma), and 47 cases of non-adenomatous polyps. According to the APCS score, the detection rate of colorectal adenomatous polyps of the high-risk group (33.3%, 33/99) was 2.02 and 3.76 times higher than those of the medium-risk group (16.5%, 39/237) and low-risk group (8.9%, 14/158), respectively (both Bonferroni correction test, both P<0.016). The detection rate of colorectal adenomatous polyps of patients with BMI>23.9 kg/m 2 was significantly higher than that of patients with BMI≤23.9 kg/m 2 (22.2%, 59/266 vs. 11.8%, 27/228), and the difference was statistically significant ( χ2=9.126, P=0.003). There was no statistically significant difference in the detection rate of colorectal adenomatous polyps between patients with positive- mSEPT9 expression and patients with negative- mSEPT9 expression (22.4%, 15/67 vs. 17.3%, 47/271) ( χ2=0.913, P=0.378). Among 158 low and medium risk patients (APCS score≤three points) who underwent simultaneous BMI measurement, FOBT and plasma mSEPT9 test, the detection rate of colorectal adenomatous polyps in patients with BMI>23.9 kg/m 2 was higher than that in patients with BMI≤23.9 kg/m 2 (17.8%, 16/90 vs. 5.9%, 4/68), and the difference was statistically significant ( χ2=4.957, P=0.030). The redetection efficacy of colorectal adenomatous polyps in patients with BMI>23.9 kg/m 2 and FOBT-positive was higher than that in patients with BMI≤23.9 kg/m 2 and FOBT-negative (28.1%, 9/32 vs. 8.0%, 4/50) and the difference was statistically significant ( χ2=5.942, P=0.027). In addition, the redetection rate of colorectal adenomatous polyps of patients with positive expression of FOBT and plasma mSEPT9 was also higher than that of patients with negative expression (5/14 vs. 12.9%, 12/93), and the difference was statistically significant ( χ2=4.738, P=0.045). Conclusions:When the APCS scoring system is used for sequential screening of colorectal tumors, the optinal choice of BMI replacement or combined with FOBT can improve the patients′ compliance and screening efficiency, which has significant clinical significance and promotion value in the early diagnosis and treatment of colorectal neoplasms.

Objective To know the anxiety status of preschool children in rural areas and to explore its influential factors. Methods Subjects of 1363 rural preschool children aged 3-6 years were selected from Anhui Province( Changfeng and Feixi county of Hefei city; Qianshan county of Anqing city; Funan county of Fuyang city) . The primary caregivers of preschool children were investigated by Chinese version of Spence Preschool Anxiety Scale (SPAS),Devereux Early Childhood Assessment for Preschoolers (DECA-P2) ,Neglect Norms for Children Aged 3 to 6 Years in Rural Areas of China,Adaptation Partnership Growth Affection Resolve (APGAR),Self-rating anxiety scale (SAS) and Self-rating depression (SDS). Result-s Among the 1363 preschool children,the positive rate of anxiety was 14. 7％. Physical injury fears score was highest (1. 24±0. 84),followed by social phobia anxiety (0. 86±0. 75) and separation anxiety (0. 85± 0. 74) . Except for obsessive compulsive disorder,the other four types of anxiety and total anxiety scores were negatively correlated with the total protective factors of mental resilience (r=-0. 054- -0. 070,P<0. 05). Besides,all anxiety scores were positively correlated with behavioral problems(r=0. 121-0. 237,P<0. 05)and neglect degree (r=0. 157-0. 269,P<0. 05). The results of multiple linear regression analysis of different anxiety types showed that gender,family type,family income status,parental rearing pattern was consistent or inconsistent,children' s neglect degree,behavior problem and anxiety/depression status of primary caregivers were the main factors affecting the anxiety level of preschool children in rural areas. Conclusions In order to reduce or avoid anxiety of preschool children,it is necessary to establish a good family environment,im-prove parents' upbringing level,and take effective education intervention.