Nipah virus (NiV) and related viruses form a distinct henipavirus genus within the Paramyxoviridae family. NiV continues to spillover into the humans causing deadly outbreaks with increasing human-bat interaction. NiV encodes the large protein (L) and phosphoprotein (P) to form the viral RNA polymerase machinery. Their sequences show limited homologies to those of non-henipavirus paramyxoviruses. We report two cryo-electron microscopy (cryo-EM) structures of the Nipah virus (NiV) polymerase L-P complex, expressed and purified in either its full-length or truncated form. The structures resolve the RNA-dependent RNA polymerase (RdRp) and polyribonucleotidyl transferase (PRNTase) domains of the L protein, as well as a tetrameric P protein bundle bound to the L-RdRp domain. L-protein C-terminal regions are unresolved, indicating flexibility. Two PRNTase domain zinc-binding sites, conserved in most Mononegavirales, are confirmed essential for NiV polymerase activity. The structures further reveal anchoring of the P protein bundle and P protein X domain (XD) linkers on L, via an interaction pattern distinct among Paramyxoviridae. These interactions facilitate binding of a P protein XD linker in the nucleotide entry channel and distinct positioning of other XD linkers. We show that the disruption of the L-P interactions reduces NiV polymerase activity. The reported structures should facilitate rational antiviral-drug discovery and provide a guide for the functional study of NiV polymerase.
Nipah Virus/chemistry* ; Cryoelectron Microscopy ; Viral Proteins/genetics* ; RNA-Dependent RNA Polymerase/genetics* ; Phosphoproteins/genetics* ; Humans ; Models, Molecular ; Protein Binding

Objective:To investigate the effect of curcumin on blood glucose levels in pregnant women at high risk of Gestational Diabetes Mellitus(GDM).Methods:One hundred and twenty-four pregnant women with high-risk factors for GDM were included in a cross-sectional study according to the inclusion and exclusion criteria.After undergoing an Oral Glucose Tolerance Test,participants were divided into the GDM group(n=61)and the non-GDM group(n=63).Subsequently,a randomized controlled trial was performed to compare Fasting Plasma Glucose(FPG)levels between the control group(n=8)and the intervention group(n=8).Results:The cross-sectional study revealed that the GDM group had significantly higher rates of pre-pregnancy BMI ≥ 24 kg/m2,early pregnancy HbA1c≥5.7％,impaired fasting glucose or glucose tolerance,and lack of exercise compared to the non-GDM group(P＜0.05).FPG levels in the GDM group were significantly higher than those in the non-GDM group at all stages of pregnancy.Multivariable linear regression analysis showed that the impact of high-risk factors on FPG gradually diminished as pregnancy progressed.In the randomized controlled experiment,all cases in the control group developed GDM;one case in the curcumin intervention group did not,whose intervention time was the earliest and longest.Pearson correlation analysis indicated a positive correlation between the duration of curcumin intervention and changes in FPG values,although the correlation was not statistically significant(P＞0.05).By the descriptive statistical analysis,within-group comparisons showed no significant differences in the median and percentiles of FPG values between the control group in late pregnancy and early pregnancy.However,the median and percentiles of FPG values were significantly lower post-intervention compared to curcumin pre-intervention.Between-group comparisons revealed that the mean FPG in the curcumin intervention group decreased significantly more than in the control group.These results suggested that curcumin might have a potential impact on FPG.No significant differences were observed in neonatal outcomes between the curcumin intervention and control groups.Conclusions:Pregnant women with high risk factors for GDM should be paid enough attention in clinical practice.All these results have demonstrated that curcumin has a positive regulatory effect on FPG in patients with GDM,which may provide a new adjunctive method for the treatment of GDM.

Objective:To investigate the effect of curcumin on blood glucose levels in pregnant women at high risk of Gestational Diabetes Mellitus(GDM).Methods:One hundred and twenty-four pregnant women with high-risk factors for GDM were included in a cross-sectional study according to the inclusion and exclusion criteria.After undergoing an Oral Glucose Tolerance Test,participants were divided into the GDM group(n=61)and the non-GDM group(n=63).Subsequently,a randomized controlled trial was performed to compare Fasting Plasma Glucose(FPG)levels between the control group(n=8)and the intervention group(n=8).Results:The cross-sectional study revealed that the GDM group had significantly higher rates of pre-pregnancy BMI ≥ 24 kg/m2,early pregnancy HbA1c≥5.7％,impaired fasting glucose or glucose tolerance,and lack of exercise compared to the non-GDM group(P＜0.05).FPG levels in the GDM group were significantly higher than those in the non-GDM group at all stages of pregnancy.Multivariable linear regression analysis showed that the impact of high-risk factors on FPG gradually diminished as pregnancy progressed.In the randomized controlled experiment,all cases in the control group developed GDM;one case in the curcumin intervention group did not,whose intervention time was the earliest and longest.Pearson correlation analysis indicated a positive correlation between the duration of curcumin intervention and changes in FPG values,although the correlation was not statistically significant(P＞0.05).By the descriptive statistical analysis,within-group comparisons showed no significant differences in the median and percentiles of FPG values between the control group in late pregnancy and early pregnancy.However,the median and percentiles of FPG values were significantly lower post-intervention compared to curcumin pre-intervention.Between-group comparisons revealed that the mean FPG in the curcumin intervention group decreased significantly more than in the control group.These results suggested that curcumin might have a potential impact on FPG.No significant differences were observed in neonatal outcomes between the curcumin intervention and control groups.Conclusions:Pregnant women with high risk factors for GDM should be paid enough attention in clinical practice.All these results have demonstrated that curcumin has a positive regulatory effect on FPG in patients with GDM,which may provide a new adjunctive method for the treatment of GDM.

OBJECTIVES: To investigate the distribution of body mass index (BMI) and risk factors for obesity in children with Diamond-Blackfan Anemia (DBA). METHODS: The children with DBA who attended National Clinical Research Center for Blood Diseases, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, from January 2003 to December 2020 were enrolled as subjects. The related clinical data and treatment regimens were recorded. The height and weight data measured within 1 week before or after follow-up time points were collected to calculate BMI. The risk factors for obesity were determined by multivariate regression analysis in children with DBA. RESULTS: A total of 129 children with DBA were enrolled, among whom there were 80 boys (62.0%) and 49 girls (38.0%), with a median age of 49 months (range 3-189 months). The prevalence rate of obesity was 14.7% (19/129). The multivariate logistic regression analysis showed that the absence of ribosomal protein gene mutation was closely associated with obesity in children with DBA (adjusted OR=3.63, 95%CI: 1.16-11.38, adjusted P=0.027). In children with glucocorticoid-dependent DBA, obesity was not associated with age of initiation of glucocorticoid therapy, duration of glucocorticoid therapy, and maintenance dose of glucocorticoids (P>0.05). CONCLUSIONS There is a high prevalence rate of obesity in children with DBA, and the absence of ribosomal protein gene mutation is closely associated with obesity in children with DBA.
Child ; Male ; Female ; Humans ; Anemia, Diamond-Blackfan/genetics* ; Pediatric Obesity/complications* ; Glucocorticoids/therapeutic use* ; Prevalence ; Risk Factors ; Ribosomal Proteins/genetics* ; Mutation

Objective: To investigate the diagnostic value of rapid antigen test based on colloidal gold immunochromatographic assay for the detection of SARS-CoV-2 infection in symptomatic patients. Methods: From May 20 to June 5 2022, 76 hospitalized children and their 55 accompanying family members with confirmed SARS-CoV-2 infection in the COVID-19 isolation unit of the Children's Hospital of Fudan University (designated referral hospital for SARS-CoV-2 infection in Shanghai) enrolled. Their nasopharyngeal swab specimens were consecutively collected. The samples were tested for SARS-CoV-2 nucleic acid by real-time quantitative. SARS-CoV-2 antigen was tested by immunochromatography. The correlation between the antigen detection results and the change of the cycle threshold (Ct) values were evaluated, as well as the sensitivity and specificity of SARS-CoV-2 antigen detection at different periods after the onset of the disease. Kappa consistency test was conducted to investigate the consistency between the 2 diagnostic methods. Results: Of the enrolled SARS-CoV-2 symptomatic infections, 76 were children, including 41 males and 35 females, with an age of 5 (2, 9) years; 55 were accompanying families, including 8 males and 47 females, with an age of 38 (32, 41) years. All 478 samples were simultaneously tested for SARS-CoV-2 antigen and nucleic acid. In any period from disease onset to negative conversion of viral nucleic acid, the overall sensitivity of the rapid antigen test was 48.2% (119/247), the specificity was 98.3% (227/231), and antigen test and nucleic acid test showed moderate consistency (κ=0.46, P<0.05). The sensitivity of antigen test was 100% (82/82) when the Ct value was ≤25. And the sensitivity of antigen test was 8/10, 4/15 and 8.3% (3/36) when the Ct value was 26, 30 and 35, respectively. All antigen tests were negative when Ct value was >35. During the period of 1-2 days, 3-5 days, 6-7 days, 8-10 days and >10 days after onset, the sensitivity and specificity of SARS-CoV-2 antigen test were 5/8 and 5/5, 90.2% (37/41) and 5/5, 88.9% (24/27) and 2/5, 45.0% (36/80) and 94.1% (32/34), 18.7% (17/91) and 98.9% (183/185) respectively. The Ct values of nasopharyngeal swabs were<26 during 2 to 7 days after onset, 28.7±5.0 on day 8, 34.5±2.9 on day 13 and > 35 after 14 days, respectively . Conclusion: SARS-CoV-2 antigen test in the patients with SARS-CoV-2 infection shows acceptable sensitivity and specificity within 7 days after onset of disease, and the sensitivity was positively correlated with viral load and negatively correlated with onset time.
Male ; Child ; Female ; Humans ; COVID-19 ; SARS-CoV-2 ; China ; COVID-19 Testing ; Nucleic Acids

Objective Obstructive sleep apnea (OSA) is closely related to obesity, insulin resistance and inflammation. Secreted frizzled-related protein 5 (SFRP5) is a recently discovered adipokine. It is involved in insulin resistance and inflammation in obesity. This study aimed at evaluating the association between SFRP5 and sleeping characteristics as well as biochemical parameters of OSA patients. Methods This was a prospective case control study. Nondiabetic OSA patients and controls were consecutively recruited and divided into three groups: OSA group, apnea-hypopnea Index (AHI)≥5/h; healthy controls with normal body mass index (BMI); obese controls without OSA, and BMI > 24.0 kg/m. All participants underwent polysomnography (PSG). Plasma SFRP5 was examined using enzyme-linked immunosorbent assay (ELISA). Blood biochemical examinations, including fasting blood glucose (FBG), lipid profile, hypersensitive C-reactive protein (hsCRP), were performed early in the morning after PSG. Patients with severe OSA were treated with nasal continuous positive airway pressure (nCPAP), and plasma SFRP5 was repeatedly measured for comparison. Results Sixty-eight subjects were enrolled in the study, including 38 patients of OSA, whose medium AHI was 58.70 /h (36.63, 71.15), 20 obese controls, and 10 healthy controls. The plasma SFRP5 level of OSA patients was not significantly different from that of healthy controls or obese controls. In OSA patients, SFRP5 level correlated positively with triglyceride level (r=0.447, P=0.005) and negatively with LDL-cholesterol level and HDL- cholesterol level (r=-0.472 and P=0.003; r=-0.478 and P=0.002; respectively). SFRP5 level was not found correlating with FBG, AHI, or any of nocturnal hypoxia parameters. After overnight nCPAP treatment, plasma SFRP5 levels of OSA patients did not change significantly (t=1.557, P = 0.148) compared to that of pretreatment. Conclusions In nondiabetic OSA patients, plasma SFRP5 is associated with the lipid profile. However, no correlation was observed between SFRP5 and FBG or sleep parameters. The SFRP5 level of OSA patients did not differ from that of non-OSA individuals in our study.

Water-soluble polysaccharides from traditional Chinese medicine have properties of complex structure and high molecular, resulting in hardly complete their structural characterization.However, a "bottom-up" approach could solve this problem.Glehniae Radix extract was extracted with hot water and then precipitated by 40% ethanol to obtain Glehniae Radix polysaccharides (RGP). Subsequently, a partial acid hydrolysis method was carried out and the effects of acid concentration, time and temperature on hydrolysis were investigated. Under the optimum hydrolysis condition (1.5 mol•L⁻¹ trifluoroacetic acid, 4 h, and 80 ℃), RGP were hydrolyzed to characteristic oligosaccharide fragments. Futher, a hydrophilic liquid chromatography- mass spectrometry method was used for the separation and structural characterization of the polysaccharide hydrolysates. According to MS and MS/MS analysis of several standard disaccharides, a method for determining the type of polysaccharide glycosidic linkage by mass spectrometry was established. The results showed that the polysaccharide hydrolysates were linear glucan containing 1, 4-glycosidic bonds. And gluco-oligosaccharides with the degrees of polymerization (DP) of 4-11 were obtained after partial acid hydrolysis.

Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.

OBJECTIVE To observe the effect of electroacupuncture on the expression of VEGF and VEGFR2 in the uterusof the rat model of ovulation induction and to investigate the possible mechanism.METHODS Female SD rats aged 10 weekswere divided into control,model and treat groups randomly,10 in each group.The model group and the treatment group wereperitoneal injected with pregnant horse serum gonadotrophin(PMSG)of 20 U and injected of the same dose of human chorionicgonadotropin(hCG)after 48 h,and then cohabitated with male SD nice until next morning.A week before cohabitation,thetreat group received a consecutive electroacupuncture treatment for 7 d(dilatational wave of 2/15 Hz for 30 min and stimulativeintensity of 1 mA).The animals were sacrificed 3.5 d after the cohabitation.The uterine tissue was taken to observe the morphologicalchange,immunehistochemical method was used to inspect the expression of VEGF and VEGFR2,and qPCR technologywas used to determine the expression of VEGF and VEGFR,with the calculation by 2-△△Ct .RESULTS Macroscopicobservation:the surface in control group was regular and smooth;the model group was intumescentia,dull-red and hyperaemia;the treatment group was slightly swollen,rosy and no congestion.The expression of VEGF and VEGFR2 were observed inthe 3 groups by immunohistochemical method,with no significant difference in average optical density,but differences in theexpression distribution.Compared with the control group,the expression distribution of stromal cells and glands in the modelgroup was more extensive,and the expression of VEGFR2 in myometrium increased significantly(P <0.05).The electroacupuncturetreatment inhibited the the expression of VEGFR2 significantly(P <0.05).The expression of VEGF mRNA in uterinetissues of model group was 0.68 times of control group,with significant declining trend;the treat group was 0.88 times ofmodel group,with significant increasing trend.The expression of VEGF2 mRNA in uterine tissues of model group was 0.46times of control group,with significant declining trend;the treat group was 0.82 times of model group,with significant increasingtrend.CONCLUSION The electroacupuncture can adjust the expression of VEGF and its receptor in the uterine tissue of ovulation induction rat model,thus regulate the synchronization of the endometrial planting window caused by hormone

Background: Pathologic response is evaluated according to the extent of tumor regression and is used to esti-mate the efficacy of preoperative treatment. Several studies have reported the association between the pathologic response and clinical outcomes of colorectal cancer patients with liver metastases who underwent hepatectomy. However, to date, no data from Chinese patients have been reported. In this study, we aimed to evaluate the asso-ciation between the pathologic response to pre-hepatectomy chemotherapy and prognosis in a cohort of Chinese patients. Patients and methods: In this retrospective study, we analyzed the data of 380 liver metastases in 159 patients. The pathologic response was evaluated according to the tumor regression grade (TRG). The prognostic role of pathologic response in recurrence-free survival (RFS) and overall survival (OS) was assessed using Kaplan–Meier curves with the log-rank test and multivariate Cox models. Factors that had potential influence on pathologic response were also analyzed using multivariate logistic regression and Kruskal–Wallis/Mann–WhitneyU tests. Results: Patients whose tumors achieved pathologic response after preoperative chemotherapy had significant longer RFS and OS than patients whose tumor had no pathologic response to chemotherapy (median RFS: 9.9 vs. 6.5 months,P= 0.009; median OS: 40.7 vs. 28.1 months,P= 0.040). Multivariate logistic regression and Kruskal–Wallis/Mann–WhitneyU tests showed that metastases with small diameter, metastases from the left-side primary tumors, and metastases from patients receiving long-duration chemotherapy had higher pathologic response rates than their control metastases (allP < 0.05). A decrease in the serum carcinoembryonic antigen (CEA) level after preopera-tive chemotherapy predicted an increased pathologic response rate (P < 0.05). Although the application of targeted therapy did not significantly influence TRG scores of all cases of metastases, the addition of cetuximab to chemother-apy resulted in a higher pathologic response rate when combined with irinotecan-based regimens rather than with oxaliplatin-based regimens. Conclusions: We found that the evaluation of pathologic response may predict the prognosis of Chinese colo-rectal cancer patients with liver metastases after preoperative chemotherapy. Small tumor diameter, long-duration chemotherapy, left primary tumor, and decreased serum CEA level after chemotherapy are associated with increased pathologic response rates.