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 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.

Objective To investigate the diagnostic value of dynamic-extended focused assessment with sonography for trauma (D-EFAST) in patients with multiple trauma in intensive care unit (ICU). Methods A prospective clinical study was conducted. Eighty patients with multiple trauma admitted to ICU of Anhui Provincial Hospital from September 1st, 2014 to December 31st, 2016 were enrolled. Extended focused assessment with sonography for trauma (E-FAST) check was conducted at first, for those who had positive findings diagnosis was confirmed by immediately CT examination or surgical exploration. If it was negative, the patients received E-FAST every morning for 7 days (defined as D-EFAST), for those with positive findings, immediately CT or surgery was performed to clarify the diagnosis. The final clinical diagnosis was used as the "gold standard" to calculate the diagnostic accordance rate of EFAST and D-EFAST examination technique for pneumothorax, pleural effusion, spleen injury, kidney damage, liver damage, gastrointestinal injury, pericardial effusion, bladder rupture, and pancreatic injury, as well as their sensitivity, specificity, positive predictive value, negative predictive value, accuracy rate, and missed diagnosis rate, and the difference between EFAST and D-EFAST was compared. Results There were 4 patients excluded because of death and abandoning treatment, and finally 76 patients were included in the study. The total sensitivity of E-FAST examination technique for pneumothorax, pleural effusion, spleen injury, liver damage, gastrointestinal injury, pericardial effusion, and bladder rupture was 75.9% (66/87), and the specificity was 98.3% (587/597), the positive predictive value was 86.8% (66/76), and the negative predictive value was 96.5% (587/608), the accuracy rate was 95.5% (653/684), and the rate of missed diagnosis was 24.1% (21/87). The most of the delayed injury in patients with multiple trauma occurred at 2-7 days after injury with incidence of 4.8% (33/684). The diagnostic sensitivity of D-EFAST for delayed injury was 98.3% (118/120), the specificity was 99.8% (563/564), the positive predictive value was 99.2% (118/119), the negative predictive value was 99.6% (563/565), the diagnostic accuracy rate was 99.6% (681/684), and rate of missed diagnosis was 1.7% (2/120). When the final clinical diagnosis was set as the "gold standard", D-EFAST technology for the detection rate was 98.3% (118/120) for patients with multiple trauma on organ injury while the detection rate of E-FAST was 75.9% (66/87), with statistical significant difference (P < 0.01), indicating that D-EFAST was better than E-FAST in check of multiple trauma patients with organ injury. Conclusion Although the E-FAST technology can quickly diagnose the multiple trauma patients and win the rescue time for critical patients, multiple trauma patients injured after 2-7 days prone to delayed damage and are difficult to detect, and D-EFAST can be used to find delayed damage earlier, and reduce the misdiagnosis rate of multiple trauma patients.

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 investigate the clinical application of skin bridge preopening window drainage in enhanced recovery of mixed hemorrhoids surgically treated with external dissection and internal ligation.Methods A total of 620 patients with mixed hemorrhoids surgically treated with external dissection and internal ligation were retrospectively analyzed from March 1,2021-February 28,2022.Patients were divided into the observation group(n=304)and the control group(n=316)according to the single and even numbers randomly generated at the time of admission.In the observation group,on the basis of conventional operation and drug treatment in the control group,most of them chose to implement preopening window drainage of the skin bridge according to the intraoperative evaluation of the anal skin bridge according to the unified evaluation criteria.Symptom-sign scoring scale used to evaluate the anal edema and pain after the operation.To compare and analyze differences in anal edema,anal pain and the postoperative hospital stay between the observation group and the control.Results The average daily edema scores of the observation group and the control group were 0.56 and 1.2 points,respectively.The average daily edema score of the observation group was lower than that of the control group,with the statistically significant differences(P＜0.05).The average daily pain scores of the observation group and the control group were 0.6 and 1.201+0.289 points,respectively.The average daily pain score of the observation group was lower than that of the control group,with the statistically significant differences(P＜0.05).The average hospitalization days of the observation group and the control group were 8 days and 10 days,respectively.The average length of stay in the observation group was less than that in the control group,with the statistically significant differences(P＜0.05).Conclusion Skin bridge preopening window drainage can effectively relieve the degree of anal edema and pain in patients with mixed hemorrhoids surgically treated with external dissection and internal ligation,improve clinical efficacy,shorten the postoperative hospital stay.It has practical value and clinical significance for enhanced recovery.

Tongue diagnosis is one of the diagnostic methods of Traditional Chinese Medicine (TCM), but its development has always been restricted by the lack of objective quantitative indicators. With the rapid development of computer technology and the advent of the algorithm era, the modernization of TCM tongue diagnosis has gradually become a hot research spot. This paper annalyses the literature and related patents of the modernization of tongue diagnosis and summarizes the R&D progress and application of tongue diagnosis as well as related instruments. It is found that domestic and foreign scholars focus on tongue diagnosis related research and attach importance to the formulation of relevant international standards. Tongue collection and analysis technology continues to develop; tongue diagnostic instruments are also gradually enriched. At present, their applications are extended to family self-use, but they are still mainly used in teaching, scientific research and other fields, involving the clinical efficacy evaluation of TCM, clinical case classification and health management, and there is still much room for development. In the future, we should strengthen the communication between multi-regional research centers, promote the communication among talents in different fields, constantly make up for the deficiencies and promote the development of tongue diagnosis research.

Objective:To evaluate the in vitro cross-neutralization of serum antibodies in human and mice immunized with inactivated SARS-CoV-2 vaccine against Delta and Beta variants. Methods:Human serum samples after a second and a third dose of inactivated SARS-CoV-2 vaccine and mouse serum samples after a two-dose vaccination were collected. The neutralizing antibodies in the samples against SARS-CoV-2 strains of prototype, Delta and Beta variants were detected using micro-neutralization assay in biosafety level Ⅲ laboratory. The seroconversion rates and geometric mean titers (GMTs) of antibodies were calculated.Results:The seroconversion rates of antibodies in human serum samples against different SARS-CoV-2 strains were all above 95%. After two-dose vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 109, 41 and 15, respectively. The GMTs decreased by 2.7 folds and 7.3 folds for the Delta and Beta variants as compared with the prototype strain. After the booster vaccination, the GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 446, 190 and 86, respectively. The GMTs of neutralizing antibodies against Delta and Beta variants decreased by 2.3 folds and 5.2 folds as compared with that against the prototype strain. The seroconversion rates of antibodies against different SARS-CoV-2 strains in mouse serum samples were all 100%. The GMTs of neutralizing antibodies against the prototype, Delta and Beta strains were 2 037, 862 and 408, respectively. The GMTs decreased by 2.4 folds and 5.0 folds for the Delta and Beta variants.Conclusions:Inactivated SARS-CoV-2 vaccine could induce a certain level of neutralizing antibodies against Delta and Beta variants in both human and mouse models. Moreover, a third dose of vaccine induced higher levels of neutralizing antibodies against Delta and Beta variants in human. This study provided valuable data for the clinical application and protective evaluation of the inactivated SARS-CoV-2 vaccine.

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.