Objective:To obtain hepatitis B virus capsid-like particles (CLPs) displaying B-cell linear epitopes of SARS-CoV-2 spike protein and evaluate their immunogenicity.Methods:Four recombinant plasmids expressing fusion proteins (M1-HBc, S1-57-HBc, S14P5-HBc and S21P2-HBc) were constructed by separately replacing codon of alanine at position 80 of hepatitis B virus core protein (HBc) with four genes coding for four B-cell linear epitopes (M1, S1-57, S14P5 and S21P2). These four recombinant proteins were expressed in E. coli BL21 Star (DE3) strains. The expression products were identified using SDS-PAGE, Western blot and native agarose gel electrophoresis (NAGE). CLPs were purified by sucrose density gradient ultracentrifugation, verified for antigenicity by Western blot and used to immunize BALB/c mice. Serum antibody titers were detected by ELISA. Results:The recombinant fusion proteins M1-HBc and S1-57-HBc self-assembled into M1-CLP and S1-57-CLP. The titer of antibody against S1-57 polypeptide in S1-57-CLP-immunized mouse serum approached 1∶1 000 000.Conclusions:Hepatitis B virus CLPs displaying SARS-CoV-2 M1 or S1-57 linear epitopes are successfully expressed in a prokaryotic system and purified. S1-57-CLP has good immunogenicity. This study provides a new idea for the development of novel diagnostic reagents and vaccines for SARS-CoV-2.

The traditional Chinese medicine （TCM） in pediatric care has a long history，proven efficacy，and distinctive characteristics.The China Association of Chinese Medicine has organized a series of youth salons to discuss the clinical advantages of treating diseases.Experts at this seminar proposed that the superior disease categories in pediatric TCM are significant for showcasing the unique strengths and advantages of TCM in the treatment of pediatric diseases，enhancing diagnostic and treatment levels，inheriting TCM knowledge，promoting the integration of TCM and Western medicine，and meeting patients' needs.The strengths of pediatric TCM are summed up as "having what others do not，excelling at what others have，being special in what others excel at，and ensuring safety in what others specialize in." The scope of superiority in pediatric TCM covers multiple systems，including respiratory，digestive，endocrine，psychological，and dermatological systems.This article summarized the advantages of TCM in treating 13 diseases discussed in the salon，such as upper respiratory tract infections，coughs，pneumonia，allergic rhinitis，bronchial asthma，atopic dermatitis，functional dyspepsia，functional constipation，enuresis，marginal short stature，simple obesity，attention deficit hyperactivity disorder，and tic disorders.The overall advantages were reflected in three aspects：First，the holistic concept and treatment based on syndrome differentiation in TCM highlight the advantage of treating the root causes of diseases，making the treatment methodical and precise.Second，most traditional Chinese herbs are natural and have strong safety profiles.Third，TCM treatment methods are abundant，especially the external treatment methods，which are widely used in pediatrics，highly accepted by parents，and have definite efficacy，as well as good safety and compliance.

Objective To observe the morphological changes of levator hiatus in patients with gestational diabetes mellitus(GDM)after spontaneous delivery with ultrasonography.Methods A total of 302 pregnant women in the first trimester(6-8 weeks)were prospectively observed,and the parameters of pelvic floor muscle hiatus were measured with ultrasound during the first trimester,also 12 weeks,6 months and 1 year after delivery at resting-state(resting period),anal contraction state(systole period)and Valsalva maneuver(tension period),respectively.Blood glucose was measured at 28 weeks of gestation,GDM was diagnosed,and the pregnant women enrolled were divided into GDM group and non-GDM group.The ultrasonic parameters and postpartum pelvic floor muscle tension were compared between groups.Spearman correlation analysis was used to evaluate the correlations of pelvic floor muscle tension grade and anal levator hiatus parameters in GDM group.Results Totally 153 pregnant women were enrolled and assigned into GDM group(n=51)and non-GDM group(n=102).Transversal diameter of levator hiatus(LH-TD),anteroposterior diameter of levator hiatus(LH-APD)and levator hiatus area(LHA)in different periods 12 weeks postpartum in both groups were higher than those in early pregnancy(all P＜0.05).Six months and 1 year postpartum,in GDM group,LH-APD and LHA in systole period,also LH-TD,LH-APD and LHA in tension period were higher than those in early pregnancy(all P＜0.05),while in non-GDM group,LH-APD and LHA in tension period were higher than those of early pregnancy(all P＜0.05).One year after delivery,LH-APD and LHA in systolic period,as well as LH-TD and LHA in tension period in GDM group were all higher than those in non-GDM group(all P＜0.05),whereas the proportion of pelvic floor muscle tension of grade Ⅱ-Ⅲ was higher,of grade Ⅳ-Ⅴ was lower in GDM group than those in non-GDM group(P＜0.05).One year after delivery,pelvic floor muscle tone grade in GDM group was negatively correlated with LH-TD,LH-APD and LHA in resting,systole and tension period(all P＜0.05).Conclusion The morphology of levator hiatus changed greatly in GDM patients after spontaneous delivery,and rehabilitation training should be carried out early.

Objective To explore the clinical significance of serum cytokine expression in the hepatitis B surface antigen(HBsAg)sero-clearance in patients with severe hepatitis B.Methods A nested case-control trial was conducted on 14 inpatients with severe hepatitis B admitted in our hospital from 2006 to 2020.Of them,7 patients(aged 36.57±3.15 years)achieved HBsAg sero-clearance within 1 year after the onset of hepatitis B flares(with abrupt rise of ALT level to＞5 times the upper limit of normal during HBV infection)and were assigned into HBsAg clearance group,while the other 7 patients(aged 34.14±2.97 years)only obtained HBsAg decreased less than 1 g within 1 year after the onset(HBsAg non-clearance group).Then,multiplex liquid-chip assay based on Luminex xMAP was used to detect the expression levels of 48 cytokines such as IFN-γ and IL-2 in serum samples of these 14 patients.Results The serum levels IFN-γ,IL-2Ra and SDF-1α were significantly lower in the HBsAg clearance group than the HBsAg non-clearance group(P＜0.05),but no statistical differences were observed in other 39 cytokines between the 2 groups.And there were 5 cytokines having no mutual expression in both groups.The copy number of HBV DNA was positively correlated with serum HGF(r=0.675,P=0.008)and SDF-1α levels(r=0.587,P=0.027),while negatively with IP-10(r=-0.600,P=0.023)and MIG level(r=-0.640,P=0.014).Meanwhile,a positive correlation was found between HBsAg titer and IL12-p70 level(r=0.593,P=0.025),and a negative correlation between HBsAg titer and TNF-α level(r=-0.609,P=0.021).In addition,the serum total bilirubin level was positively correlated with the expression of SCGF-β(r=0.543,P=0.045).Conclusion Three differentially expressed cytokines and some cytokines related to HBV DNA level and HBsAg titer are found,which may provide new insights into the underlying immunological mechanism of HBV virus clearance caused by hepatitis flares.Meanwhile,it also provides potential biomarkers for HBsAg serological clearance in patients with severe hepatitis B.

OBJECTIVES: This work aimed to investigate the molecular mechanism of cyclic tensile stress (CTS) stimulating autophagy in human periodontal ligament cells (hPDLCs). METHODS: hPDLCs were isolated and cultured from normal periodontal tissues. hPDLCs were loaded with tensile stress by force four-point bending extender to simulate the autophagy of hPDLCs induced by orthodontic force du-ring orthodontic tooth movement. XMU-MP-1 was used to inhibit the Hippo signaling pathway to explore the role of the Hippo-YAP signaling pathway in activating hPDLC autophagy by tensile stress. The expression levels of autophagy-related genes (Beclin-1, LC3, and p62) in hPDLCs were detected by real-time quantitative polymerase chain reaction. Western blot was used to detect the expression levels of autophagy-related proteins (Beclin-1, LC3-Ⅱ/LC3-Ⅰ, and p62) and Hippo-YAP pathway proteins (active-YAP and p-YAP) in hPDLCs. Immunofluorescence was used to locate autophagy-related proteins (LC3-Ⅱand p62) and Hippo-YAP pathway proteins (active-YAP) of hPDLCs. RESULTS: CTS-activated autophagy in hPDLCs and expression of autophagy-related proteins initially increased and then decreased; it began to increase at 30 min, peaked at 3 h, and decreased (P<0.05). CTS increased the expression of active-YAP protein and decreased the expression of p-YAP protein (P<0.05). When XMU-MP-1 inhibited the Hippo-YAP signaling pathway (P<0.05), active-YAP protein was promoted to enter the nucleus and autophagy expression was enhanced (P<0.05). CONCLUSIONS The Hippo-YAP signaling pathway is involved in the regulation of autophagy activation in hPDLCs under CTS.
Humans ; Hippo Signaling Pathway ; Periodontal Ligament/metabolism* ; Beclin-1/metabolism* ; Cells, Cultured ; Autophagy

Objective:To investigate the correlation between Helicobacter pylori ( H.pylori) infection and albuminuria inphysical examination population. Methods:It was a cross-sectional study. All the adults who received physical examination and underwent 13C-urea breath test at the Physical Examination Center in Tongji Hospital in 2021 were selected as the study subjects. General data (such as demographic information and past medical history) were documented. The physical measurement and blood biochemical indicators were checked too. Multivariate analysis was used to analyze the relationship between H. pylori infection and albuminuria. Results:A total of 30 311 subjects were included in this analysis. There were 17 123 males and 13 188 females with an age of (44.51±12.17) years. The positive rate of H. pylori infection was 27.3%. The incidence of albuminuria in subjects with H. pylori infection was 6.7%, and it was 6.1% in the subjects without H. pylori infection ( P=0.031). After adjusting for the confounding factors such as gender, age, diabetes and hypertension, H. pylori infection was independently associated with the risk of albuminuria (odds ratio ( OR)=1.133, 95% CI: 1.018-1.261, P=0.022). Conclusion:H. pylori infection is positively correlated with the occurrence of albuminuria in the physical examination population.

We aimed to evaluate the effectiveness and safety of single-course initial regimens in patients with low-risk gestational trophoblastic neoplasia (GTN). In this trial (NCT01823315), 276 patients were analyzed. Patients were allocated to three initiated regimens: single-course methotrexate (MTX), single-course MTX + dactinomycin (ACTD), and multi-course MTX (control arm). The primary endpoint was the complete remission (CR) rate by initial drug(s). The primary CR rate was 64.4% with multi-course MTX in the control arm. For the single-course MTX arm, the CR rate was 35.8% by one course; it increased to 59.3% after subsequent multi-course MTX, with non-inferiority to the control (difference -5.1%,95% confidence interval (CI) -19.4% to 9.2%, P = 0.014). After further treatment with multi-course ACTD, the CR rate (93.3%) was similar to that of the control (95.2%, P = 0.577). For the single-course MTX + ACTD arm, the CR rate was 46.7% by one course, which increased to 89.1% after subsequent multi-course, with non-inferiority (difference 24.7%, 95% CI 12.8%-36.6%, P < 0.001) to the control. It was similar to the CR rate by MTX and further ACTD in the control arm (89.1% vs. 95.2%, P =0.135). Four patients experienced recurrence, with no death, during the 2-year follow-up. We demonstrated that chemotherapy initiation with single-course MTX may be an alternative regimen for patients with low-risk GTN.
Antineoplastic Combined Chemotherapy Protocols/adverse effects* ; Dactinomycin/adverse effects* ; Female ; Gestational Trophoblastic Disease/drug therapy* ; Humans ; Methotrexate/therapeutic use* ; Pregnancy ; Retrospective Studies

Objective:To investigate the relationship between liver function and arterial stiffness in the physical examination population.Methods:A total of 9 111 people who received physical examination in the Health Management Center of Wuhan Tongji Hospital from January to December 2017 with an age of 18 and above and complete clinical data were included in this study. The subjects were divided into arterial stiffness group (3 252 cases) and the normal group (5 859 cases) according to the results of brachial-ankle artery pulse wave velocity (baPWV) examination. The liver function indicators were checked in all the subjects, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and glutamyl transpeptidase γ (GGT). According to interquartile range of the liver function indicators, the subjects were divided into 4 groups (group Q1- Q4). Multivariate logistic regression analysis and restricted cubic spline functions were used to explore the relationship between liver function and risk of arterial stiffness. Results:When the subjects were grouped by ALT, after adjusted for age and gender, the risk of arterial stiffness in the Q2 to Q4 groups was 1.260 (95% CI:1.093-1.452, P<0.05), 1.571 (95% CI:1.355-1.822, P<0.001) and 2.436 (95% CI:2.097-2.830, P<0.001) times of that in the Q1 group, respectively, P for trend<0.001. And after adjusted for age, gender, body mass index, low-density lipoprotein cholesterol (LDL-C), triglycerides, high-density lipoprotein cholesterol (HDL-C), the risk of arterial stiffness in the Q2 to Q4 groups was 1.158 (95% CI:1.003-1.338, P<0.05), 1.331 (95% CI:1.143-1.551, P<0.001) and 1.867 (95% CI:1.591-2.190, P<0.001) times of that in the Q1 group, respectively, P for trend <0.001. After adjusted for age, gender, body mass index, LDL-C, triglycerides, HDL-C, systolic blood pressure, diastolic blood pressure, heart rate, fasting blood glucose, blood uric acid, serum creatinine, with or without hypertension or diabetes, the risk of arterial stiffness in the Q2 to Q4 groups was 1.116 (95% CI:0.940-1.325, P=0.210), 1.241 (95% CI:1.036-1.488, P<0.05) and 1.598 (95% CI:1.322-1.932, P<0.001) times of that in the Q1 group, respectively, P for trend <0.001. Restricted cubic spline regression analysis showed a linear positive correlation between ALT and the risk of arterial stiffness. Similar results as ALT obtained with AST, ALP and GGT. Conclusions:The increase of liver function indicators is positively correlated with the risk of arterial stiffness.

Objective:To analyse the relationship between serum electrolyte concentrations and risk of cardiovascular events in physical examination population.Methods:A cross-sectional study design was applied to survey 8 445 adults whose serum high-sensitivity cardiac tropon Ⅰ (hs-cTnⅠ) and serum electrolytes (chloride, phosphorus, calcium, sodium, potassium and magnesium) concentrations were measured at the health examination center of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 1, 2018 to February 28, 2022. The risk of cardiovascular events was classified into three levels according to the serum hypersensitive cardiac troponin Ⅰ(hs-cTnⅠ) concentration: low, middle or high risk group. One-way analysis of variance was applied to compare the differences in serum electrolyte concentrations of participants with different risk levels of cardiovascular events. Ordered multi-category logistic regression was performed to analyze the correlation between serum electrolyte levels and the risk of cardiovascular events.Results:The concentration of potassium and magnesium ion in the subjects with low risk of cardiovascular events were both higher than those in the middle and high risk group [potassium ion (4.28±0.29) vs (4.24±0.34), (4.23±0.36) mmol/L, magnesium ion (0.88±0.06) vs (0.87±0.07), (0.87±0.07) mmol/L](both P<0.05), while the concentration of sodium ion was lower [(140.54±1.75) vs (140.88±1.73), (140.81±2.20) mmol/L]( P<0.001); the concentration of phosphorus ion in the high-risk group was lower than those in the middle and low risk groups [(1.04±0.17) vs (1.08±0.16), (1.05±0.15) mmol/L]( P=0.001); no significant difference was found in the concentrations of chloride and calcium ion among the three groups (both P>0.05). Compared to subjects with normal concentrations of electrolyte, the risk level of cardiovascular events in subjects with hypokalemia ( OR=6.96, 95% CI: 3.67-13.10) and hypomagnesemia ( OR=5.00, 95% CI: 1.01-24.50) was higher(both P<0.05). Within the normal range, sodium concentration was positively correlated with the risk of cardiovascular events ( OR=1.08, 95% CI: 1.03-1.14; P<0.001). Conclusions:The serum sodium, potassium and magnesium concentrations in health examination subjects are correlated with the risk of cardiovascular events. Maintaining the balanced concentration of serum potassium and magnesium, as well as low sodium levels within normal limits may help prevent cardiovascular events.

Neonatal sepsis is a common infectious disease in the neonatal period, and its morbidity and mortality are increasing year by year.Its etiology and pathogenesis have not been fully elucidated.In recent years, studies have confirmed that plasma soluble CD14 subtype(sCD14-ST)plays a certain role in the pathogenesis of neonatal sepsis, and has a certain value in its diagnosis and prevention.The study found that sCD14-ST could be used as an indicator for early auxiliary diagnosis of neonatal sepsis, and the expression of sCD14-ST was positively correlated with the degree of neonatal sepsis.Early detection of sCD14-ST can predict neonatal sepsis.This article reviews the research on sCD14-ST and its application in neonatal sepsis.