Objective To select polyethylenimine polymer with proper molecular weight to enhance the gene expression efficiency of Adenovirus (Ad).Methods Ad was resepcitively complexed with PEI-600,PEI-1 800 and PEI 25 000 by electrostatic adsorption.Afterwards,MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay was used to investigate the cytotoxicity of the complexes with a serial concentration,followed by cell transduction assay to select the best amount of PEI with fixed dose of Ad.In the further,the physicochemical characteristics of the optimized complexes,Zeta potential,and size distribution were studied.Results MTT showed the higher the PEI moderate molecular weight was,the greater the cytotoxity was.PEI-25 000 showed a greater cytotoxicity than that of PEI-600 and PEI-1 800.As compared with PEI-600 and PEI-25 000,PEI-1 800 significanlty promoted expression of Ad gene in A549 cells.PEI turned the Zeta potential of Ad from negative to positive charge.Conclusion In the three kinds of polymer/adenovirus compounds,the cytotoxicity of the compound is positively correlated with the polymer molecular weight and concentration.The greater molecular weight the polymer as well as the higher the concentration are,the higher the cytotoxicity is.

Objective To investigate the effect of miRNA-200b-specific inhibitor on hepatic stellate cells(HSCs) activation,proliferation,and extracellular matrix production.Methods The miRNA-200b-specific inhibitors were designed,synthesized,and transfected into HSCs with lipofectamine 2000.The supernatant and HSCs were collected after incubation for 48 h.The expression of miR-200b was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR).The expression ofα-smooth muscle actin (oα-SMA) protein in HSCs was detected by Western blotting.The cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT) method.Contents of type Ⅲ procollagen and hyaluronic acid in supernatant were determined by radioimmunoassay.Results Compared to the control group,miRNA-200b expression was decreased in the miRNA-200b inhibitor group by 82％ (P ＜ 0.01),α-SMA protein expression was reduced in the miRNA-200b inhibitor group by (19 ± 3) ％ (P ＜ 0.05),and the activity of HSCs proliferation was reduced by(33 ± 5)％ (P ＜0.01),and the contents of type Ⅲ procollagen and hyaluronic acid in supernatant were reduced in miRNA-200b inhibitor group by (35 ± 4)％ and (31 ± 2)％,respectively(P ＜0.01).Conclusions The miRNA-200b-specific inhibitor could significantly reduce the expression of miRNA-200b,and inhibit HSC proliferation,activation,and extracellular matrix production.

Objective To prepare berberine chitosan sustained-release tablets and to study the in vitro release characteristics. Methods The chitosan sustained-release tablets were prepared by chitosan as the skeleton material,and the drug release rate of berberine hydrochloride at different time was determined by high performance liquid chromatography. Results The in vitro cumulative release of berberine chitosan sustained-release tablets was increased with time.Furthermore,it can sustain for 24 h and the accumulative release was above 95%, the dissolution time of 63. 2% of the drug was 12. 63 h. Artificial gastric juice had the best performance on the dissolution of berberine,while rotations only influenced the dissolution at the beginning of drug release. Conclusion This method is convenient, accurate and reproducible.Berberine chitosan sustained-release tablets have further development and application value.

Objective:To investigate the risk factors for anastomotic leakage after laparo-scopic lower anterior resection (LAR) of rectal cancer, and the application value of its risk assess-ment scoring model.Methods:The retrospective case-control study was conducted. The clinico-pathological data of 539 patients who underwent laparoscopic LAR of rectal cancer in 13 medical centers, including 248 cases in Renji Hospital of Shanghai Jiaotong University School of Medicine, 35 cases in Ningbo First Hospital, 35 cases in Changzhou Second People's Hospital, 32 cases in the First People's Hospital of Nantong, 32 cases in Linyi People's Hospital, 31 cases in Changzhou Wujin People's Hospital, 28 cases in Jiading District Hospital of Traditional Chinese Medicine, 27 cases in the First Hospital of Taizhou, 26 cases in Shanghai Pudong Gongli Hospital, 21 cases in the People's Hospital of Rugao, 11 cases in Central Hospital of Fengxian District, 7 cases in Ningbo Hangzhou Bay Hospital and 6 cases in Jiangsu jianhu People's Hospital, from January 2016 to November 2020 were collected. There were 157 males and 382 females, aged (62.7±0.5)years. Observation indicators: (1) follow-up; (2) risk factors for anastomotic leakage after laparoscopic LAR; (3) establishment of risk assessment scoring model for anastomotic leakage after laparoscopic LAR. Follow-up was conducted by outpatient examination or telephone interview. Patients were followed up at 1 week after discharge or 1 month after the operation to detect the anastomotic leakage. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were represented as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the chi-square test and multivariate analysis was conducted usong the Logistic regression model. The area under curve of receiver operating characteristic curve was used to estimate the efficiency of detecton methods. The maximum value of the Youden index was defined as the best cut-off value. Results:(1) Follow-up: 539 patients were followed up at postoperative 1 week and 1 month. During the follow-up, 79 patient had anastomotic leakage, with an incidence of 14.66%(79/539). Of the 79 patients, 39 cases were cured after conservative treatment, 40 cases were cured after reoperation (ileostomy or colostomy). (2) Risk factors for anastomotic leakage after laparoscopic LAR. Results of univariate analysis showed that sex, age, body mass index, smoking and/or drinking, tumor diameter, diabetes mellitus, hemoglobin, albumin, grade of American Society of Anesthesio-logists (ASA), neoadjuvant chemoradiotherapy, distance from anastomotic level to dentate line, the number of pelvic stapler, reinforced anastomosis, volume of intraoperative blood loss, placement of decompression tube, preservation of left colic artery, operation time and professional doctors were related factors for anastomotic leakage after laparoscopic LAR ( χ2=14.060, 4.387, 5.039, 4.094, 17.488, 33.485, 25.066, 28.959, 34.973, 34.207, 22.076, 13.208, 16.440, 17.708, 17.260, 4.573, 5.919, 5.389, P<0.05). Results of multivariate analysis showed that male, tumor diameter ≥3.5 cm, diabetes mellitus, hemoglobin <90 g/L, albumin <30 g/L, grade of ASA ≥Ⅲ, neoadjuvant chemoradiotherapy, distance from anastomotic level to dentate line <1 cm, the number of pelvic stapler ≥3, non-reinforced anastomosis, volume of intraoperative blood loss ≥100 mL and no placement of decom-pression tube were independent risk factors for anastomotic leakage after laparoscopic LAR ( odds ratio=2.864,3.043,12.556,7.178,8.425,12.895,8.987,4.002,3.084,4.393,3.266,3.224,95% confidence interval as 1.279?6.411, 1.404?6.594, 4.469?35.274, 2.648?19.459, 2.471?28.733, 4.027?41.289, 3.702?21.777, 1.746?9.171, 1.365?6.966, 1.914?10.083, 1.434?7.441, 1.321?7.867, P<0.05). (3) Establishment of risk assessment scoring model for anastomotic leakage after laparoscopic LAR. based on the results of univariate analysis, clinicopathological factors with χ2>20, χ2>10 and ≤20 or χ2≤10 were defined as scoring of 3, 2, 1, respectively. The cumulative clinicopatho-logical factors scoring ≥6 was defined as an effective evaluating indicator for postoperative anastomotic leakage. The risk assessment scoring model (6-321) for anastomotic leakage after laparoscopic LAR was established. The cumulative value ≥6 indicated high incidence of anastomotic leakage, and the cumulative value <6 indicated low incidence of anastomotic leakage. Conclusions:Male, tumor diameter ≥3.5 cm, diabetes mellitus, hemoglobin <90 g/L, albumin <30 g/L, grade of ASA ≥Ⅲ, neo-adjuvant chemoradiotherapy, distance from anastomotic level to dentate line <1 cm, the number of pelvic stapler ≥3, non-reinforced anastomosis, volume of intraoperative blood loss ≥100 mL and no placement of decompression tube are independent risk factors for anastomotic leakage after laparoscopic LAR. The risk assessment scoring model (6-321) is established according to the above results.The cumulative value ≥6 indicates high incidence of anastomotic leakage and the cumulative value <6 indicates low incidence of anastomotic leakage.

Background Chloropropanol esters (MCPDE) have attracted much attention in recent years as a kind of new contaminant found in various refined oils. The pollution of 3-monochloropropane-1,2-diol esters (3-MCPDE) is the most prominent. 3-MCPDE is hydrolyzed in organisms to 3-monochloropropane-1,2-diol which has been shown to have reproductive toxicity, nephrotoxicity, and potential carcinogenicity. Infant formula powders can be polluted by 3-MCPDE when refined edible oils are added during production. Objective To assess the risk of exposure to 3-MCPDE related to the consumption of infant formula powders for children aged 0-3 years in Shanghai market using the food consumption data and the data of 3-MCPDE contamination in these foods. Methods Gas chromatography-mass spectrometry was used to determine 3-MCPDE in 133 samples of infant formula powders in Shanghai. Using a multistage stratified random sampling method, a total of 807 infants and toddlers aged 0-3 years were randomly selected from each district/rural/town in Shanghai, including 208 children aged 0-6 months, 276 children aged 6-12 months, and 323 children aged 12-36 months. The food consumption data was investigated using food frequency questionnaire (FFQ) distributed by combining telephone inquiries and face-to-face interviews among guardians. Population's probability distributions of 3-MCPDE exposure were assessed by @Risk 7.5 software and Monte Carlo simulation algorithm. According to the dietary intake assessment model and the tolerable daily intake (TDI) of 3-MCPDE of 2.00 μg·(kg·d)⁻¹ proposed by the European Food Safety Authority (ESFA), a risk assessment of exposure to 3-MCPDE was conducted for infants and toddlers aged 0 to 3 years old in Shanghai who consumed 3-MCPDE via infant formula powders. Results The average concentration of 3-MCPDE in 133 samples of infant formula powders was 0.115 mg·kg⁻¹ with a positive rate of 100.00%. Among different types of formula powders, infant formula powders for infants of 0-6 months had the highest concentrations of 3-MCPDE and fat [0.136 mg·kg⁻¹ and 25.2 g (per 100 g sample) in average respectively]. There was a positive correlation between fat concentration and 3-MCPDE concentration in the samples (r=0.438, P<0.05). The average consumption of infant formula powders of 807 infants aged 0-3 years was 88.3 g·d⁻¹. Among all investigated age groups, the average consumption of infant formula powders by infants aged 0-6, 6-12, and 12-36 months was 87.7, 98.3, and 80.1 g·d⁻¹ respectively. The dietary exposure to 3-MCPDE from infant formula powders of infants aged 0-3 years averaged 0.83 μg·(kg·d)⁻¹ for general intake level or valued 1.44 μg·(kg·d)⁻¹ using the 95 percentile for high intake level. Exposure decreased with increasing age and was highest in infants 0-6 months of age, whose general and high intake levels were 1.41 and 2.34 μg·(kg·d)⁻¹, respectively. The risk population defined with the exposure higher than the TDI proposed by EFSA were proportioned to be 13.90% and 0.50% in infants aged 0-6 months and 6-12 months respectively, indicating a risk that cannot be ignored, and no risk in infants aged 12-36 months. Conclusion Among the investigated infants aged 0-3 years in Shanghai, those aged 0-6 months are at a high risk of exposure to 3-MCPDE. In view of the sensitivity of infants to pollutant exposure, the risk of exposure to 3-MCPDE should be highly concerned.

Background: Esophageal mucosal injury induced by gastroesophageal reflux is a key link to the development of Barrett's esophagus-associated adenocarcinoma. However, the molecular mechanism is still not elucidated. Aims: To investigate the role of differentially expressed genes (DEGs) after stimulating esophageal cells with acid and bile acid in the development of esophageal adenocarcinoma (EAC). Methods: The DEGs were obtained through bioinformatics methods after stimulating esophageal cells with low pH and deoxycholic acid, and GO, KEGG enrichment analysis were performed. Protein-protein interaction (PPI) network was performed to screen the hub genes, and their relationships with prognosis and tumor stage of EAC patients were analyzed. The role of co-expressed genes of GADD45B in EAC was also analyzed. Results: Thirty-one overlapping DEGs were obtained after stimulating esophageal cells with low pH and deoxycholic acid, which mainly enriched in the cytokine-cytokine receptor interaction, transcription factors activity, and regulation of cell proliferation and apoptotic process. High expression of GADD45B was correlated with the survival prognosis and tumor stage of EAC patients. GADD45B and its co-expressed genes were involved in the production of tumor necrosis factor. Conclusions: The high expression of GADD45B induced by acid and bile acid is correlated with the prognosis and tumor stage of EAC patients, and is a potential diagnosis and treatment target for Barrett's esophagus-associated adenocarcinoma.

ObjectiveTo conduct comprehensive assessment of internal and external cadmium exposure and health risks for Shanghai residents. MethodsCadmium levels in food samples were calculated by employing two dietary exposure assessment methods， total diet study （TDS） and food frequency questionnaire （FFQ）， to estimate the daily dietary cadmium exposure of Shanghai residents. The provisional tolerable monthly intake （PTMI） of cadmium set by joint food and agriculture organization/WHO expert committee on food additives （JECFA） was applied to evaluate the health risk. Differences in dietary and urinary cadmium were compared by rank-sum test among different regions， age， gender， smoking status， and BMI groups， and the association between internal and external cadmium exposure was investigated by correlation analysis. ResultsThe mean value of urinary cadmium for 1 300 respondents was 0.542 μg·L^-1. Urinary cadmium was higher in the population in central urban and urban-rural fringe areas than in the suburban area， higher in the older age group than in the younger age group， and higher in the smoking group than in the non-smoking group （all P<0.01）. The two assessment methods showed that the mean values of daily dietary cadmium exposure for Shanghai residents were 0.306 and 0.090 μg·kg^-1， with 3.69% and 0.85% of Shanghai residents exceeding the PTMI， respectively. Correlation analyses showed that dietary exposure to cadmium based on the FFQ method was positively correlated with the urinary cadmium level when smoking status， age， gender， and BMI were adjusted. ConclusionDietary exposure to cadmium of Shanghai residents is mainly derived from vegetables， aquatic products， cereals and potatoes， and is overall at a low-risk level. Dietary exposure assessment based on FFQ and risk monitoring data can effectively estimate long-term cadmium exposure.

ObjectiveTo explore the influence of the core symptoms of attention deficit hyperactivity disorder （ADHD） on behavioral problems of children with ADHD propensity， so as to provide references for early identification and targeted intervention for children with ADHD propensity. MethodsFrom July to August 2021， 25 children with ADHD propensity were screened as the ADHD propensity group， and 25 children matched for age， gender and grade were included as the normal group in an elementary school in Guangzhou. ADHD core symptoms were assessed by the Chinese version of the Swanson Nolan and Pelham， version IV-parent form for ADHD （SNAP-IV）， and behavioral problems were assessed by Questionnaire-Children with Difficulties （QCD） and Conners Parental Symptom Questionnaire （PSQ）. Spearman correlation analysis was used to examine the correlation between ADHD core symptoms and QCD and PSQ scores， and hierarchical linear regression analysis was used to explore the effect of ADHD core symptoms on behavioral problems. Results① The differences between the groups showed that both attention deficit and hyperactivity-impulsivity factor scores were higher in the ADHD propensity group than those in the normal group （t=7.771， 6.726， P<0.01）. ② Correlation analysis showed that the attention deficit factor score was negatively correlated with QCD total score （r=-0.440， P<0.05）， and positively correlated with the learning problem factor score of PSQ （r=0.457， P<0.05）. The score of hyperactivity-impulsivity was negatively correlated with score of anxiety factor in PSQ （r=-0.457， P<0.05）， and positively correlated with impulse-hyperactivity factor score （r=0.552， P<0.01）. ③ Hierarchical linear regression analysis showed that the attention deficit factor score negatively predicted the total score of QCD （B=-0.682， P<0.05， R²=0.468）. The hyperactivity-impulsivity factor score had a negative predictive effect on the anxiety factor score of PSQ （B=-0.048， P<0.05， R²=0.367）， and had a positive predictive effect on the impulsivity-hyperactivity factor score （B=0.077， P<0.01， R²=0.424）. ConclusionChildren with ADHD propensity have significant attention deficit symptoms， hyperactivity-impulsivity symptoms and behavioral problems， and the attention deficit may be the main cause of their daily behavioral problems， while hyperactivity-impulsivity may be the main cause of their impulsive-hyperactivity problems.