Human astrovirus (HastV) is recognized as one of the leading causes of acute viral diarrhea in infants. The HastV non-structural protein, nsPla, and C-terminal protein, nsPla/4, contain various conserved functional domains,and may play an important role in virus replication, transcription and the virus-host interactions of HastV. This study used an E. coli system to investigate the expression of nsPla and nsPla/4 proteins. Firstly,the nsPla and nsPla/4 genes of HAstV-1 were cloned into the prokaryotic expression vector,PGEX-4T-1, to build the PGEX-4T-1a and PGEX-4T-la/4 fusion protein plasmids. Then, the recombinant plasmids were transformed into Escherichia coli BL21 (DE3) and induced with isopropyl-β-D-thiogalactopyranoside (IPTG). The optimal expression conditions of the two fusion proteins were identified and then analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting, respectively. The results showed that the pGEX-4T-la fusion protein was maximally expressed at 30 °C after 12 hours of induction with 1.0 mM IPTG. The pGEX-4T-la/4 fusion protein was maximally expressed at 20 °C after 8 hours of induction with 0.5 mM IPTG. Western blot analysis showed that the two fusion proteins specificity reacted with the anti-nsPla and anti-GST monoclonal antibodies, respectively. This study successfully obtained the HAstV non-structural protein, nsP1a, and its C-terminal protein nsP1a/4 protein using an E. coli system. This novel study lays the foundation for future research into the pathogenic mechanisms of human astrovirus and the functions of its non-structural protein.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Mamastrovirus ; genetics ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

Mycoplasma pneumoniae is one of the main cause of community-acquired pneumonia in children.The incidence of mycoplasma pneumoniae pneumonia has increased in recent years,and some of them are severe cases.Children who infected with mycoplasma pneumonia can present multi-system damage including nervous system,hematological system,digestive system and so on,in addition to respiratory symptoms.Children who suffered from mycoplasma pneumoniae pneumonia was more reported to present embolism or thrombosis in children,and some of them even develop disability and mortality.This review summarized the recent references on the thrombophilia presented in children who suffered from mycoplasma pneumoniae pneumonia,including the mechanisms,indicators and treatments,so that pediatricians can diagnose and treat early to prevent the serious complications and improve the children's quality of life.

Objective:To investigate the correlation between thyroid function index and serum visfatin in patients with acute pancreatitis (AP).Methods:Using a prospective design, 65 patients with AP treated in Binzhou People's Hospital from December 2017 to December 2019 were selected as the research subjects. They were divided into mild acute pancreatitis (MAP) group ( n = 35) and severe acute pancreatitis (SAP) group ( n = 30) according to the acute physiology and chronic health Ⅱ (APACHE Ⅱ) score ( < 8, ≥8 scores). At the same time, 40 healthy people were selected as the control group. The serum levels of free thyroxine (FT 4), free triiodothyronine (FT 3) and thyrotropin (TSH) were measured by chemiluminescence immunoassay. Radioimmunoassay was used to measure serum reverse triiodothyronine (rT 3) level, and enzyme-linked immunosorbent assay was used to measure serum visfatin level. Pearson method was used to analyze the correlation between thyroid function index and serum visfatin in patients with AP. Results:The FT 4 [(14.02 ± 3.63), (15.68 ± 3.05) pmol/L], FT 3 [(2.34 ± 0.80), (3.66 ± 0.65) pmol/L], and TSH levels [(2.78 ± 0.85), (3.10 ± 0.57) mU/L] in SAP and MAP groups were significantly lower than those in control group [(17.03 ± 3.96), (6.04 ± 1.55) pmol/L, (4.88 ± 2.30) mU/L, P < 0.05], but the rT 3 levels [(1.63 ± 1.12), (1.23 ± 0.26) nmol/L] were significantly higher than that in control group [(0.97 ± 0.28) nmol/L, P < 0.05]. There was significant difference in serum FT 3 levels between SAP and MAP groups ( P < 0.05). The serum visfatin levels of SAP, MAP and control groups were (10.75 ± 2.92), (3.70 ± 1.73), (2.30 ± 1.31) ng/ml, the difference between the three groups was statistically significant ( F = 67.174, P < 0.05). The serum visfatin levels in SAP and MAP groups were higher than that in control group, and that in SAP group was higher than that in MAP group ( P < 0.05). There were negative correlations between serum FT 3 level and visfatin level in SAP and MAP groups ( r = - 0.672, - 0.610, P < 0.05). Conclusions:The serum visfatin level and thyroid function index of AP patients are abnormal. The levels of FT 3 and visfatin are correlated with the severity of AP patients, and there is a negative correlation between the levels of FT 3 and visfatin. Detection of thyroid function index is helpful to judge the AP patient's condition.

Objective To investigate the effect of periodontal mechanical treatment on serum interleukin-6 (IL-6) and carotid artery matrix metalloproteinase(MMP)-2 and MMP-9 expression in chronic periodontitis(CP) SD rats with atherosclerosis (As).Methods Forty-four six-week-old male SD rats were randomly divided into three groups:control group(group A),As group (group B),As + CP group (group C).According to different periodontal interventions,group C was randomly subdivided into four groups:natural process group (C 1),the periodontal mechanical treatment group (C2),the periodontal mechanical treatment + local drugs group(C3),and the periodontal mechanical treatment + local and system drugs group(C4).Each group received the appropriate treatment and periodontal interventions.Serum IL-6 levels were determined by enzyme linked immunosorrbent assay (ELISA).MMP-2 and MMP-9 levels in the proximal aorta were examined by immunohistochemistry.Results The gray value of MMP-2 and MMP-9 was basically the same in all groups.Compared with group A,the gray value of MMP-2 and MMP-9 of group B and C were decreased.C1 group showed the formation of atherosclerotic plaque and fibrous cap.Compared with group B (126.4 ± 2.0,124.8 ± 2.8),the gray value of group C1 (101.3 ± 2.4,101.2 ± 4.1) was significantly weaker(P ＜ 0.05).The staining depth of MMP-2 and MMP-9 of groups C1,C2,C3 and C4 were sequentially decreased,and the differences of gray value were statistically significant(P ＜ 0.05).The levels of serum IL-6 in groups B and C1 increased gradually with time and became significantly higher than that of group A(P ＜ 0.01).The levels of serum IL-6 in groups C2,C3,and C4 increased gradually and reached the peak 5 weeks after the establishment of model (P ＜ 0.001).After that,the levels of serum IL-6 decreased gradually and was lower than baseline.The levels of serum IL-6 in groups C3 and C4 were significantly lower than that in group C2 7 weeks after the establishment of model(P ＜ 0.01).Conclusions In rats with periodontitis and cardiovascular diseases,chronic periodontal inflammation may significantly increase the severity of As and promote the formation of atherosclerotic plaque.Mechanical periodontal therapy may cause short-term systemic inflammation and then reduce vascular inflammation in long term.With supplement use of local and systemic antibiotics,the mechanical periodontal therapy may get the vascular disease and systemic inflammation improved.

Objective To establish chronic periodontitis model in SD rats,and to investigate the effect of oral intervention on atherosclerosis.Methods Fifty male SD rats were randomly divided into three groups,group A(normal control),group B(atherosclerosis,As) and group C(chronic periodontitis,CP).Group C was further divided into group C1(natural process),group C2(simple mechanical treatment),group C3 (systemic antibiotics),group C4-1(teeth extraction) and group C4-2(teeth extraction+systemic antibiotics),each group consisted of 7 rats.Every group received oral intervention.Serum interleukin(IL)-6 levels were detected in five different time points(1,3,5,7,9 weeks after a successful modeling) by enzyme linked immunosorbent assay.All animals were killed after 24 weeks.Matrix metalloproteinase(MMP)-2,9 in the proximal aorta was detected by immuno histochemistry.Results The levels of serum IL-6 in groups B and C1 increased gradually with time and became significantly higher than that in group A(P＜0.01).Levels of serum IL-6 were increased gradually in each intervention group(C2,C3,C4-1,C4-2) and reached its peak at 5 weeks after modeling[C2:(62.3 ± 14.3) ng/L,C3:(58.2±8.7) ng/L,C4-1:(127.0±29.9) ng/L,C4-2:(120.6± 23.1) ng/L].Compared with group B,group C4-1 and C4-2 increased most significantly(P＜0.01).Levels of serum IL-6 decreased gradually.Eventually,group C2[(28.6± 8.1) ng/L],C3[(40.8 ± 15.1) ng/L] and C4-2 [(32.7± 11.1) ng/L] were significantly lower than group B(P＜0.05),and in group C2 IL-6 was the lowest.Although levels of serum of IL-6 significantly decreased in group C4-1[(72.8± 16.4) ng/L],but remained the highest.Immunohistochemistry showed that MMP-2,9 were expressed in group B,C 1 and C4-1,and significantly higher than in group A(183.0±2.0,181.3 ± 2.0),the gray value differences were statistically significant(P＜0.01).Group C4-1 (123.1 ±2.9,121.0± 3.2) was the strongest,group B (126.4±2.0,124.8 ±2.8)and C1 (140.0±2.2,139.7±3.2) were decreased (P＜0.01).While group C2(169.3±3.4,169.7±2.3),C3 (149.0±1.7,145.1±2.5) and C4-2(157.7± 1.2,155.8±2.7) were significantly lower than group C1(P＜0.01),and group C2 was close to normal.Conclusions Periodontitis could increase the risk of atherosclerosis in rats with chronic periodontitis.Periodontal mechanical treatment and teeth extraction may increase the risk of As in the short time.However,the risk would gradually reduce in a long time.

Objective:To explore the mediating effect of emotional exhaustion of nurses on occupational stress and recovery experience, so as to provide reference for clinical nursing management.Methods:From June to August 2019, clinical registered nurses of the First BethuneHospital of Jilin University were selected as the research objects by convenient sampling method. Occupational Stress Inventory (OSI-R) , emotional exhaustion subscale of Maslach Burnout Inventory-General Survey (MBI-GS) and Recovery Experience Questionnaire (REQ) were used to investigate and analyze the correlation between occupational stress, emotional exhaustion and recovery experience of clinical nurses. In this study, a total of 560 questionnaires were issued and 538 valid questionnaires were recovered, with an effective recovery rate of 96.1%.Results:The total OSI-R score of 538 clinical nurses was (350.60±68.80) , the emotional exhaustion subscale score of MBI-GS was (22.65±7.05) and the total score of REQ was (51.16±12.20) . The occupational task, individual stress response and individual coping resource dimension scores of clinical nurses in REQ were all correlated with the total score of emotional exhaustion subscale ( P<0.05) , but negatively correlated with the total score of REQ ( P<0.05) . The relationship model of occupational stress, emotional exhaustion and experience recovery showed that occupational stress of clinical nurses had direct and indirect effects on experience recovery. Conclusions:There is a certain correlation between occupational stress, emotional exhaustion and recovery experience of nurses, and emotional exhaustion plays a mediating role between occupational stress and recovery experience. Nursing managers should provide a good working environment for clinical nurses, reduce occupational stress of clinical nurses, and reduce the level of emotional exhaustion to improve recovery experience of nurses.

Objective: To develop the Adolescent Mental Health Literacy Assessment Questionnaire (AMHLAQ), and to evaluate its reliability and validity among undergraduates. Methods: On the basis of the definition of mental health literacy (MHL) and the Knowledge, Attitudes and Practices (KAP) theory, this study constructed a total of 36 items consisting of four dimensions, and scores were measured according to a five point Likert type scale. Using a cluster sampling method, a questionnaire survey was conducted among 3 826 freshmen and sophomore students from two medical schools in Anhui Province. The items were screened by performing t tests, Pearson s correlation coefficient analysis and factor analysis. The reliability and validity of the questionnaire were evaluated using indicators including homogeneity reliability, the split half reliability coefficient, and construct validity. Results: Factor analysis revealed that the AMHLAQ consisted of 22 questions grouped into four domains. The variance cumulative contribution rate was 62.213%. The reliability result showed that the Cronbach s alpha coefficient of the total questionnaire was 0.897, the split half reliability was 0.800, the Cronbach s coefficient of each dimension was 0.796 to 0.885, the split half reliability of each dimension was 0.725 to 0.846, and the indicators had a high level of reliability. Confirmatory factor analysis showed that the model fit was good ( χ 2/df =19.319, P <0.01; RMSEA=0.069). Conclusion AMHLAQ is consistent with the evaluation standard of psychometrics, has good reliability and validity, and can be used to estimate the level of MHL among undergraduates.

Objective: To explore the association between middle school students health literacy and bully victimization and associated sex difference, and to provide guidance for bully prevention in adolescents. Methods: During November 2017 to January 2018, 18 900 junior and senior high school students were enrolled from Hefei of Anhui Province, Shenyang of Liaoning Province, Yangjiang of Guangdong Province and Chongqing by multistage cluster sampling. A self rated questionnaires were used to collect demographic information, health literacy, and bully victimization. Group differences by different characteristics and the association between health literacy and bully victimization were analyzed. Results: The detection rates of campus and cyber bullying victimization were 15.8% (2 992/18 900) and 9.1% (1 723/18 900). Boys, junior high school students, students with poor family financial status and few close friends had a detection rate of 28.0%, 18.8%, 23.3%, and 33.6% of school bullying, which are higher than those of the control group ( χ 2=225.64, 148.07, 141.13, 143.49, P <0.01); boys, students with poor household income and few close friends, the detection rates of cyber bullying were 10.9%, 14.4%, and 20.1%, respectively, are higher than the control group ( χ 2=62.96, 112.82, 88.49, P <0.01). Multivariate Logistic regression analysis indicated that students with low overall and dimensions scores of health literacy were more likely to suffer from campus and cyber bullying, except for the dimension of physical activity. In addition, at all levels of health literacy, males are more likely to be bullied than females ( P <0.05). Conclusion Health literacy of middle school students is related to bully victimization, which is sex specific. Intervention programs of bullying should focus on health literacy enhancement.

Objective: To explore the association between health literacy and drinking behaviors among middle school students in six provinces of China, and to provide scientific basis for the prevention and intervention of drinking behaviors. Methods: From November 2015 to January 2016, 22 628 students were enrolled from Ulanqab, Shenyang, Xinxiang, Bengbu, Chongqing and Yangjiang by multistage stratified cluster sampling method. And a questionnaire survey was conducted among them to collect demographic information, health literacy and drinking behaviors. Rates on drinking behaviors were compared in students with specific characteristics. Multiple Logistic regression analyses were conducted to examine the association between health literacy and drinking behaviors. Results: The proportion of ever drinking, current drinking, binge drinking, drunkenness was 60.0%(13 580), 16.8%(3 799), 5.9%(1 344) and 9.9%(2 250), respectively. After adjusting for gender, grade, family address, siblings, accommodation type, parental educational level, and self reported family income, compared to the high level health literacy, the low level health literacy was significantly associated with ever drinking (OR=1.76, 95%CI=1.62-1.91), current drinking(OR=1.37, 95%CI=1.23-1.52), binge drinking (OR=1.39, 95%CI=1.18-1.63), and drunkenness(OR=1.68, 95%CI=1.47-1.92), respectively(P<0.01). Conclusion Health literacy of middle school students was related to drinking behaviors. The occurrence of health risk behaviors such as drinking can be prevented and reduced by improving students health literacy level.

Objective: To observe the expression of retinal proteins in experimental autoimmune uveitis (EAU) mice and to explore the possible molecular mechanism of autoimmune uveitis. Methods: Twelve female C57BL/6J mice were randomly divided into model group and normal control group, 6 mice in each group.In the model group, the EAU model was established by subcutaneous injection of human interphotoreceptor retinoid-binding protein (IRBP) 651-670.The fundal change of EAV mice was assessed by direct ophthalmoscope, OCT and histopathological staining.At 18 days after immunization, the retinas of the two groups were taken for retinal protein extraction, protein restriction enzyme digestion, mass spectrometry detection, data analysis, and bioinformatics analysis.This study was approved by the experimental animal Ethics Committee of Tianjin Medical University Eye Hospital (TJYY2018070113). The feeding and use of experimental animals follow the ARVO statement. Results: The EAU mouse model was successfully established.At 10 days after immunitation, the retina of EAV mouse was damaged.At 18 days after immunization, retinal edema and infiltration of inflammatory cells into vitreous were observed.Proteomic results showed that a total of 4 458 proteins were identified in this study, of which 522 were differentially-expressed proteins (fold change>1.5, P<0.05). Among these differentially-expressed proteins, 147 proteins including Stat1 and Stat3 were up-regulated and 375 proteins including Gucy2f and Rho were down-regulated.These differentially-expressed proteins were closely related to platelet activation, integrin signaling, T cell activation, the phototransduction cascade, Toll-like receptor and so on. Conclusions EAU is related to the abnormal expression of Stat1, Stat3 and other proteins, as well as the abnormal regulation of platelet activation and integrin signal pathway.