As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Liver is a unique organ with polyploidy.About 20 to 50 percent of hepatocytes in adult human are polyploid cells,which contain more than two sets of chromosomes.Hepatocytes polyploidization is triggered by the changes in insulin signal during weaning,and regulated by various cell cycle regulator genes to ensure the polyploid cells proportion,ploidy and zonation.The regulation of polyploid hepatocytes is crucial for liver's metabolic and regenerative function,and protecting liver from tumorigenesis.However,in chronic virus hepatitis and nonalcoholic fatty liver disease,the pathological hepatocytes polyploidization can be found in disease progression because of cell cycle checkpoint inhibition and oxidative stress.Distinguishing physiological and pathological hepatocyte polyploidization will be helpful for the understanding the relationship between chronic liver disease and tumorigenesis.

Vibrio parahaemolyticus, the main pathogen causing seafood related food poisoning worldwide, has strong biofilm formation ability. ToxR is a membrane binding regulatory protein, which has regulatory effect on biofilm formation of V. parahaemolyticus, but the specific mechanism has not been reported. c-di-GMP is an important second messenger in bacteria and is involved in regulating a variety of bacterial behaviors including biofilm formation. In this study, we investigated the regulation of ToxR on c-di-GMP metabolism in V. parahaemolyticus. Intracellular c-di-GMP in the wild type (WT) and toxR mutant (ΔtoxR) strains were extracted by ultrasonication, and the concentrations of c-di-GMP were then determined by enzyme linked immunosorbent assay (ELISA). Three c-di-GMP metabolism-related genes scrA, scrG and vpa0198 were selected as the target genes. Quantitative real-time PCR (q-PCR) was employed to calculate the transcriptional variation of each target gene between WT and ΔtoxR strains. The regulatory DNA region of each target gene was cloned into the pHR309 plasmid harboring a promoterless lacZ gene. The recombinant plasmid was subsequently transferred into WT and ΔtoxR strains to detect the β-galactosidase activity in the cellular extracts. The recombinant lacZ plasmid containing each of the target gene was also transferred into E. coli 100λpir strain harboring the pBAD33 plasmid or the recombinant pBAD33-toxR to test whether ToxR could regulate the expression of the target gene in a heterologous host. The regulatory DNA region of each target gene was amplified by PCR, and the over-expressed His-ToxR was purified. The electrophoretic mobility shift assay (EMSA) was applied to verify whether His-ToxR directly bound to the target promoter region. ELISA results showed that the intracellular c-di-GMP level significantly enhanced in ΔtoxR strain relative to that in WT strain, suggesting that ToxR inhibited the production of c-di-GMP in V. parahaemolyticus. qPCR results showed that the mRNA levels of scrA, scrG and vpa0198 significantly increased in ΔtoxR strain relative to those in WT strain, suggesting that ToxR repressed the transcription of scrA, scrG and vpa0198. lacZ fusion assay showed that ToxR was able to repress the promoter activities of scrA, scrG and vpa0198 in both V. parahaemolyticus and E. coli 100λpir. EMSA results showed that His-ToxR was able to bind to the regulatory DNA regions of scrA and scrG, but not to the regulatory DNA region of vpa0198. In conclusion, ToxR inhibited the production of c-di-GMP in V. parahaemolyticus via directly regulating the transcription of enzyme genes associated with c-di-GMP metabolism, which would be beneficial for V. parahaemolyticus to precisely control bacterial behaviors including biofilm formation.
Vibrio parahaemolyticus/metabolism* ; Escherichia coli/metabolism* ; Bacterial Proteins/metabolism* ; Transcription Factors/genetics* ; Gene Expression Regulation, Bacterial

Lateral epicondylitis is a common clinical disease with characteristics of lateral elbow pain, insidious onset and easy recurrence, which can cause forearm pain and decreased wrist strength, seriously affecting patients′ daily life and work. Although there are various treatment methods for lateral epicondylitis with different effects, standard treatments are still lacking nowadays. Platelet-rich plasma (PRP) has good effects on bone and tendon repair, and is now widely used in the treatment of lateral epicondylitis. However, there is a lack of a unified understanding of the technology and specifications of PRP in the treatment of lateral epicondylitis. Therefore, the Sports Medicine Branch of the Chinese Medical Association and Physical Medicine and Rehabilitation Branch of the Chinese Medical Association organized experts in the fields of sports medicine and rehabilitation medicine in China to formulate the "clinical expert consensus on platelet-rich plasma treatment for lateral epicondylitis (2022 version)", and proposed suggestions based on evidence-based medicine mainly from the concept, epidemiology and pathophysiology of lateral epicondylitis, symptoms, signs and imaging manifestations of lateral epicondylitis, PRP concept and application component requirements, quality control of PRP preparation technology, indications and contraindications of PRP in the treatment of lateral epicondylitis, PRP injection in the treatment of lateral epicondylitis, application of PRP in the operation of lateral epicondylitis, related problems after PRP treatment of lateral epicondylitis, evaluation of the results after PRP treatment of lateral epicondylitis, and health and economic evaluation of PRP treatment of lateral epicondylitis, so as to provide guidance for clinical diagnosis and treatment.

Objective:To study the transcriptional regulation of pilABCD by the master quorum sensing (QS) regulator OpaR in Vibrio parahaemolyticus. Methods:Total RNAs were extracted from the wild type (WT) and opaR mutant (Δ opaR) strain. Quantitative real-time PCR (qPCR) was employed to calculate the transcriptional variation of pilA (the first gene of pilABCD operon) between WT and Δ opaR. The regulatory DNA region of pilABCD was cloned into the corresponding restriction endonuclease sites of pHRP309 harboring a promoterless lacZ reporter gene. The recombinant pHRP309 plasmid was then transferred into WT and Δ opaR, respectively, to detect the β-galactosidase activity in cellular extracts using a β-Galactosidase Enzyme Assay System (Promega). The primer extension assay was applied to map the transcription start site of pilABCD using the total RNAs extracted from the WT strain as the template. The regulatory DNA region of pilABCD was amplified by PCR, and the over-expressed His-OpaR was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns (Amersham). Thereafter, the electrophoretic mobility shift assay (EMSA) was applied to analyze the DNA-binding activity of His-OpaR to the target DNA in vitro, and the DNase I footprinting assay was further employed to detect the DNA-binding sites of His-OpaR within the target DNA. Results:The results of qPCR and LacZ fusion assays showed that OpaR activated the transcription of pilABCD, leading to a gradual increase in the expression level of pilA with the extension of culture time. The primer extension assay detected only one transcription start site located at 155 bp upstream of pilA. The results of EMSA and DNase Ⅰ footprinting assays showed that His-OpaR protected two DNA regions located from -246 to -197 bp and -181 to -131 bp upstream of pilA. Conclusions:Vibrio parahaemolyticus OpaR activated the transcription of pilABCD in a direct manner.

Objective To evaluate the impact of total parenteral nutrition(TPN)on nutrition status and inflammatory markers in hospitalized fasted patients with inflammatory bowel disease(IBD).Methods A retrospective study was performed and 82 hospitalized fasted IBD patients [male/female=58/24,(39.4±14.5)years] who received TPN entered the study.Among them,38 patients had ulcerative colitis(UC)and 44 patients suffered from Crohn`s disease(CD).Clinical data(gender,age,duration of disease,history of disease,prednisone,immuno-suppressor,and antibiotics)were obtained from medical records.Nutritional parameters,C-creative protein(CRP),and erythrocyte sedimentation rate(ESR)before and after TPN were also obtained.Average caloric supplementation by TPN was(4 437.3±1 199.1)kJ/d and the nitrogen amount was(9.9±1.7)g/d.Median PN length was 15 days(7-54 days).67 IBD patients received a TPN formula with glutamine(≥14 d,25 patients vs.0-14 d,42 patients)and 15 IBD subjects received TPN without glutamine.Malnutrition was diagnosed by body mass index(BMI)and serum albumin level.Results The prevalence of undernutrition was 90.2％(74/82)in the study population.CD patients had a significantly longer history of disease [84(3-288)months vs.24(1-324)months,P<0.001] and a significantly lower BMI [(15.6±1.8)kg/m2 vs.(19.1±3.5)kg/m2,P<0.001] compared with those in UC patients.TPN improved nutritional parameters [serum albumin:(28.7±6.6)g/L before TPN vs.(31.7±5.8)g/L after TPN,P<0.001;pre-albumin:(174.1±85.5)mg/L before TPN vs.(227.2±82.8)mg/L after TPN,P<0.001].Conclusions TPN improves nutritional status in hospitalized fasted IBD patients.However,prospective randomized controlled trials are required to estimate the role of low-to-middle dosage of glutamine in IBD patients.

Objective To investigate the relationship between serum ferritin and nonalcoholic fatty liver diseases in obese children.Methods Obese children aged 6 to 14 years old were enrolled.Duration of obesity, anthropometric parameters (height, body weight, waist circumference, hip circumference), bioelectrical impedance analysis (body fat), serological parameters (liver transaminases, lipid metabolism, fasting blood glucose, fasting insulin, serum ferritin) and liver ultrasonography were recorded.Insulin resistance (IR) index was calculated by homeostasis model assessment (HOMA).All subjects were divided into 3 groups according to liver ultrasound and liver transaminases : simple obese children (SOC) group, obese children with nonalcoholic simple fatty liver (NAFL) group and obese children with nonalcoholic steatohepatitis (NASH) group.Results 86 obese children entered the study, with a mean age of (10.4 ± 1.9) years, including 26 in the SOC group, 28 in the NAFL group and 32 in the NASH group.Waist circumference standard deviation score (SDS or Z-score), waist-to-hip ratio, HOMA-IR index and serum ferritin in the NASH group were obviously higher than those in the NAFL group [2.3 ± 0.3 vs.2.1 ± 0.3, P =0.020;1.0 ± 0.0 vs.0.9 ± 0.1,P=0.014;4.0±1.7 vs.2.9±1.8, P=0.006;(104.1 ±49.6) μg/Lvs.(68.4 ±22.7) μg/L, P=0.004] and the SOC group [2.3 ±0.3 vs.1.9 ±0.3, P=0.000;1.0±0.0vs.0.9 ±0.1, P=0.012;4.0 ±1.7 vs.2.5 ±1.6, P=0.001;(104.1 ±49.6) μg/Lvs.(59.2 ±28.9) μg/L, P=0.001], while there was no significant difference in body mass index Z-score [2.8 ± 0.5 vs.2.7 ± 0.6, P =0.524;2.8 ± 0.5 vs.2.7 ± 0.6, P =0.662].There were no significant differences between the NAFL group and the SOC group in the above indicators [2.1 ±0.3 vs.1.9 ±0.3, P =0.260;0.9 ±0.1 vs.0.9 ±0.1, P =0.952;2.9 ± 1.8vs.2.5±1.6, P=0.283;(68.4±22.7) μg/Lvs.(59.2±28.9) μg/L, P=0.161].Mter controlling age, body mass index, waist circumference, waist-to-hip ratio, triglyceride, and HOMA-IR index, serum ferritin was still positively correlated with the magnitude of nonalcoholic fatty liver diseases in obese children (r =0.335, P =0.002).Conclusion Serum ferritin is probably an independent risk factor for NASH in obese children.

Objective To evaluate the relationship between birth weight and nonalcoholic fatty liver diseases (NAFLD) in Chinese primary school students. Methods A cross-section study was conducted in ifve elementary schools in Gao Hang Town, Shanghai and 2163 students were enrolled in the study (1120 boys/1043 girls). Height, body weight, waist circumference and per-cent of body fat (bioelectrical impedance analysis) were measured by professional nutritionist after training. Birth weight, feeding pattern, height and body weight of parents were obtained by a self-completed questionnaire. NAFLD was diagnosed by ultra-sound. The risk factors of NAFLD were analyzed. Results The prevalence of NAFLD in the study population was 8.9%. The prevalence of NAFLD was signiifcantly higher in boys than that in girls (12.5%vs 5.0%, P<0.01). Logistic regression showed that sex (OR=1.97, 95%CI:1.21-3.21) and percent of body fat (OR=1.12, 95%CI:1.07-1.17) were the risk factors of NAFLD, and normal BMI was the protective factor of NAFLD (OR=0.09, 95%CI:0.04-0.19) in the study population. Conclusions The pre-valence of NAFLD is higher in boys than that in girls. Also overweight, and high percent of body fat are risk factors of NAFLD in children.

Objective To investigate the risk factors of central venous catheters (CVC) colonization.Methods A retrospective study was performed on adult patients with CVCs placement in Renji Hospital,Shanghai Jiaotong University from January 2006 to March 2010.Clinical data,catheter-related information (including duration of catheter placement,position and purpose of catheterization,and whether or not out-of-ward catheterization),catheter culture results,and prevalence of catheter-related blood stream infection (CRBSI)was collected.Results A total of 651 patients aged 18 to 97 years (median:63 years) were enrolled in the study,in whom 762 CVC were placed.The median duration of catheter placement was 1 1 days (2 to 122 days)and the total duration of CVC placement was 10 725 days.The prevalence of catheter colonization was 16％(122/762),and 134 germs were cultured.Gram-positive cocci was the most common colonized bactera (52.2％,70/122),followed by gram-negative bacilli (33.6％,45/122) and fungi (14.2％,19/122).Overall 13 CRBSI were confirmed and the rate of CRBSI was 1.21/1000 catheter-days.Logistic regression analysis demonstrated that the risk factors for CVC colonization included mechanical ventilation [odds ratio (OR) =1.783,95％ confidence interval (Cl) =1.108 ～2.870],serum albumin concentration less than 25 g/L before catheterization (OR =1.783,95％ Cl =1.357 ～ 6.757),prolonged duration of catheter placement (OR =1.105,95％ Cl =1.009 ～ 1.111),and out-of-ward catheterization (OR =2.837,95％ Cl =1.010 ～7.969).Conclusion Patients with prolonged duration of catheter placement and out-of-ward catheterization are inclined to CVC colonization.

Objectives To investigate the prevalence of non-alcoholic fatty liver (NAFLD) disease and its correlation with chronic metabolic diseases in two primary school students in Shanghai. Methods One thousand ifve hundred and thirty-two 7-11 year-old students from two primary schools were enrolled in Septamber-October 2011. The anthropometric indices, blood pressure, screening for pseudoacanthosis nigricans and liver ultrasonography of all subjects were recorded. Results The overall prevalence of NAFLD, obesity, abdominal obesity, pseudoacanthosis nigricans, high systolic blood pressure and high diastolic blood pressure was 6.5%, 26.7%, 16.3%, 5.1%, 1.7%and 1.9%, respectively. The prevalence of NAFLD and abdomi-nal obesity in students of central urban area was signiifcantly higher than that in suburban area (P<0.01). The binary regression analysis revealed a signiifcant association between NAFLD in students with sex, age, obesity, abdominal obesity, pseudoacan-thosis nigricans and high systolic blood pressure (P<0.05). Conclusions NAFLD has close correlation with chronic metabolic disease in children. It is time to adopt prevention, detection and treatment in NAFLD children with symptoms of chronic meta-bolic diseases.