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 explore the levels of Kruppel like factor 5(KLF5)and Dickkopf related protein 1(DKK1)in the serum of patients with primary liver cancer and analyze their diagnostic value.Methods A to-tal of 98 patients with primary liver cancer diagnosed in the hospital from March 2021 to March 2024 were in-cluded as the experimental group,and they were divided into early group(40 cases)and late group(58 cases)according to the severity of the disease.Meantime,101 patients with benign liver disease admitted to the hos-pital during the same period time were selected as the control group,and 100 healthy individuals who under-went the physical examination during the same period were selected as the healthy group.Serum samples were obtained from the groups and general clinical data were collected and organized.Enzyme linked immunosor-bent assay was applied to detect the levels of KLF5 and DKK1,and the differences and changes in serum KLF5 and DKK1 levels among the groups were analyzed.Multivariate Logistic regression was applied to ana-lyze the influencing factors of occurrence of primary liver cancer.Receiver operating characteristic curve was used to analyze the efficacy of serum KLF5 and DKK1 levels in the diagnosis of primary liver cancer.Results The levels of alpha fetoprotein(AFP),carcinoembryonic antigen(CEA),KLF5 and DKK1 in the ex-perimental group were significantly higher than those in control group and healthy group(P＜0.05).The lev-els of serum KLF5 and DKK1 were obviously different in patients with primary liver cancer with or without lymph node metastasis(P＜0.05).The levels of serum KLF5 and DKK1 in late group were higher than those in early group(P＜0.05).AFP,CEA,KLF5 and DKK1 levels were independent risk factors for primary liver cancer(P＜0.05).The area under the curve of the combined diagnosis of serum KLF5 and serum DKK1 was 0.928,with specificity of 93.10％,which was better than that of the individual diagnosis(Z combination-DKK1=2.186,Z combination-KLF5=3.386,P=0.029,P=0.001).Conclusion The levels of KLF5 and DKK1 in the serum of patients with primary liver cancer are obviously elevated and change with changes in the condition of dis-ease.The combined detection of KLF5 and DKK1 is of great significance for the diagnosis of primary liver cancer.

Objective:To understand the molecular characteristics of the mutant strain of polymerase basic protein 2 (PB2) gene of influenza A (H1N1pdm) in Guangdong province, and to explore its specific molecular sites, so as to provide scientific basis for the prevention and control of influenza virus.Methods:Throat swab samples were collected from 2 cases infected with PB2 gene variant strains for virus isolation, and 23 influenza virus strains were selected from Guangdong province for sequencing analysis. The reference sequences and vaccine strain sequences provided by GISAID were used to perform evolutionary analysis on hemagglutinin (HA) and PB2 genes. Virus strain antigen analysis and neuraminidase (NA) inhibition test were carried out. PB2 protein model was constructed and polymerase activity was analyzed.Results:H399N amino acid mutation occurred in the HA gene of PB2-D701N and PB2-A271S variant strains, both of which belonged to the branch of 6B.1A.5a.2a. They belonged to the same big branch and different small branches as the vaccine strain A/Victoria/4897/2022, and they are all vaccine-like strains. In the three-dimensional structure, the mutations of PB2-D701N and PB2-A271S change charge and hydrophobicity.Conclusions:PB2-D701 and A271 were highly conserved, and PB2 mutant strains were not the dominant strains. The PB2 mutant had high antigenicity with the vaccine. The PB2 mutant strain is sensitive to NA inhibitors. The three-dimensional model predicted that PB2-D701N mutation could enhance virulence and affect transmissibility of influenza virus, while PB2-A271S mutation could affect polymerase activity and polymerase complex synthesis of influenza virus.

AIM:The study aimed to explore the effect of curcumin(Cur)on lipopolysaccharide(LPS)-in-duced RAW264.7 cell-derived osteoclasts,together with its underlying mechanism.METHODS:An osteoclast model was established by treating RAW264.7 cells with LPS.The viability of the cells was assessed by CCK-8 assays and osteo-clast formation was evaluated by tartrate-resistant acid phosphatase(TRAP)activity.The levels of reactive oxygen species(ROS),Fe2+,glutathione(GSH),and malondialdehyde(MDA)were examined by biochemical assays.Mitochondrial morphology was assessed by transmission electron microscopy.The mRNA and protein levels of p53,glutathione peroxi-dase 4(GPX4),and solute carrier family 7 member 11(SLC7A11)were determined by RT-qPCR and Western blot,re-spectively.RESULTS:Treatment with LPS successfully induced osteoclasts formation in RAW264.7 cells.The TRAP results showed that compared with the LPS-treated group,the number of osteoclasts and TRAP activity in the curcumin-treated group decreased dose-dependently(P<0.01).Compared with the control group,the LPS+Erastin group showed significantly increased TRAP activity(P<0.01),while after curcumin treatment,the TRAP activity declined in a dose-de-pendent manner(P<0.01).The results of the biochemical tests showed that compared with the control group,the LPS+Erastin group had significantly elevated levels of ROS,Fe2+,and MDA,while the GSH level was significantly reduced(P<0.01),and compared with the LPS+Erastin group,the ROS,Fe2+,and MDA levels in the curcumin group decreased(P<0.01)and GSH levels increased(P<0.01).These effects were all dose-dependent.Transmission electron microscopy showed that compared with the LPS group,the LPS+Erastin group had reduced mitochondrial cristae and increased mem-brane density,while after treatment with curcumin,both these effects were reversed.The RT-qPCR and Western blot re-sults showed that compared with the control group,the mRNA and protein levels of p53 in the LPS+Erastin group were sig-nificantly increased,while those of of SLC7A11 and GPX4 were significantly reduced(P<0.01).After curcumin treat-ment,the p53 mRNA and protein levels were reduced while the levels of SLC7A11 and GPX4 were increased(P<0.01).CONCLUSION:Curcumin can inhibit lipopolysaccharide-induced differentiation of RAW264.7 cells into osteoclasts,and its mechanism may be related to the inhibition of the p53/SLC7A11/GPX4 signaling pathway.

BACKGROUND: Biliary tract carcinomas (BTCs) are relatively rare but lethal primary malignant tumors derived from the biliary tract system. The burden of BTCs varies according to sex, age, region, and country, but limited attention has been paid to the burden of BTCs. We sought to explore the up-to-date data from the Global Burden of Disease Study (GBD) and expand findings by accessing the demographic features of BTC disease burden. METHODS: Using the latest data from the GBD 2021, we evaluated and analyzed the distributions and patterns of BTC disease burden in various age groups, sexes, regions, and countries. RESULTS: The number of incident cases, deaths, and disability-adjusted life-years (DALYs) tended to increase and peaked at 216,770 (95% uncertainty interval [UI]: 181,890-245,240), 171,960 (95% UI: 142,350-194,240), and 3,732,100 (95% UI: 3,102,900-4,317,000) person-years, respectively, in 2021. However, the average global age-standardized rates (ASRs) of incident cases, deaths, and DALYs shrunk by -11.46% (95% UI: -21.91 to 3.35%), -24.09% (95% UI: -33.19 to 16.88%), and -26.25% (95% UI: -35.53 to 18.36%), respectively, from 1990 to 2021. Meanwhile, the male/female ratio (male per 100 female) of incidence, deaths, and DALYs changed from 76.40, 75.41, and 74.72 to 86.89, 79.11, and 82.29, respectively. In 2021, the highest number of incident cases, deaths, and DALYs occurred in East Asia. The top three highest incidences, deaths, and DALYs were observed in China, India, and Japan, and the highest ASRs were observed in Chile in 2021. Analysis of the Human Development Index along with disease burden estimates of BTCs also suggests that the burden of the disease is related to the level of comprehensive development of the society. CONCLUSION This study provided a comprehensive comparison of differences in the burden of disease across populations and over time, and further presented evidence concerning the formulation of prevention and control policies and etiologic studies for BTCs and proposed logical hypotheses to investigate.
Humans ; Global Burden of Disease ; Biliary Tract Neoplasms/epidemiology* ; Male ; Female ; Disability-Adjusted Life Years ; Middle Aged ; Aged ; Adult ; Incidence ; Aged, 80 and over ; Quality-Adjusted Life Years ; Cost of Illness

Objective To explore the risk factors for developing chronic pulmonary heart disease in patients with pneumoconiosis.Methods The medical records of pneumoconiosis patients admitted to an occupational disease hospital in Sichuan Province between January 2012 and November 2021 were collected.Kaplan-Meier(K-M)method,or product-limit method,was used to plot the incidence curves of pulmonary heart disease in the pneumoconiosis patients.Cox proportional hazard regression model was used to analyze the influencing factors associated with chronic pulmonary heart disease in patients with pneumoconiosis.Results A total of 885 pneumoconiosis patients were included in this study.The follow-up time was 12 to 115 months and the median follow-up time was 43 months.A total of 138 patients developed chronic pulmonary heart disease and the incidence density of pulmonary heart disease was 38.50/1000 person-years.Multivariate Cox proportional hazard regression analysis showed that the influencing factors of pneumoconiosis inpatients developing chronic pulmonary heart disease included the following,being 50 and older(hazard ratio[HR]=1.85,95%confidence interval[CI]:1.25-2.74),stage Ⅲ pneumoconiosis(HR=2.43,95%CI:1.48-4.01),resting heart rate≥100 beats/min(HR=2.62,95%CI:1.63-4.21),the complication of chronic obstructive pulmonary disease(COPD)(HR=4.52,95%CI:2.12-9.63),underweight(HR=2.40,95%CI:1.48-3.87),overweight and obesity(HR=0.54,95%CI:0.34-0.86),and triacylglycerol(TG)(HR=0.69,95%CI:0.49-0.99).Conclusion Old age,stage Ⅲ pneumoconiosis,high resting heart rate,low BMI,and the complication of COPD are risk factors for chronic pulmonary heart disease in pneumoconiosis patients,while overweight and obesity and TG are protective factors.Early identification of the risk factors and the adoption of the corresponding prevention measures are the key to preventing chronic pulmonary heart disease in patients with pneumoconiosis.

Surgical operation is the main treatment method for pancreatic cancer, and in clinical practice, radical surgery for pancreatic cancer is often combined with superior mesenteric-portal vein confluence pancreaticoduodenectomy to achieve R0 resection. However, severe left-sided portal hypertension (LSPH) may occur after splenic vein dissection, resulting in a series of pathological changes such as congestive splenomegaly, thrombocytopenia, backflow obstruction of splenic vein, and gastrointestinal varices, and in some cases, it can lead to fatal gastrointestinal hemorrhage and hemorrhagic shock. Therefore, in order to better manage LSPH in clinical practice, this article systematically analyzes and reviews the pathogenesis, treatment regimens, and control strategies of LSPH after combined superior mesenteric-portal vein confluence pancreaticoduodenectomy and put forward corresponding suggestions based on current studies.

Objective:To translate the health literacy-sensitive communication scale into Chinese and examine its reliability and validity.Methods:Following Brislin′s translation principles, the HL-COM was translated and culturally adapted into Chinese. This cross-sectional study surveyed 434 outpatients and inpatients from three tertiary hospitals in Hubei Province using a questionnaire. Cronbach′s α coefficient and split half reliability were used to evaluate the reliability of the scale. Scale-level content validity index (S-CVI) and item-level content validity index (I-CVI) were used to evaluate the content validity. Exploratory factor analysis and confirmatory factor analysis were used to test the structural validity, while measurement equivalence across gender was examined.Results:The Chinese version of HL-COM contains 9 items, which was consistent with the original English version. Reliability of the scale: Cronbach′s ɑ Coefficient=0.938, Spearman-Brown half coefficient=0.926; Content validity: Scale-level content validity index (S-CVI)=0.926, Item-level content validity index (I-CVI) was 0.833-1.000; only one factor was extracted based on exploratory factor analysis, with a cumulative variance contribution rate of 68.541%. Confirmatory factor analysis indicated a good fit, with the fitting indexes of ?χ2/df=2.794, Normed Fit Index (NFI)=0.974, Standardized Root Mean Square Residual (SRMR)=0.025, Goodness-of-Fit Index (GFI)=0.962, Root Mean Square Error of Approximation (RMSEA)=0.089, Comparative Fit Index (CFI)=0.983. Multigroup confirmatory factor analysis indicated measurement invariance of the Chinese version of HL-COM across gender. Conclusion:The Chinese version of HL-COM demonstrates good reliability and validity, and serves as a valuable tool for assessing health literate in health care organization in China.

Objective:To analyze and reveal the genetic evolution and variation of influenza viruses in cases of co-infection in Guangdong Province.Methods:Throat swab samples were collected from four cases of H1N1pdm and H3N2 co-infection for viral isolation. The isolated strains were subjected to antigen analysis and neuraminidase inhibitor susceptibility test. High-throughput sequencing was used to detect the sequences of strains in three throat swab samples and one virus strain, and then genetic variations were analyzed.Results:Four influenza viruses were isolated with one strain of H1N1pdm and three of H3N2 subtype, and all of them were genetically similar to the vaccine strain in 2022-2023. The HA genes of H1N1pdm and H3N2 strains belonged to clade 6B.1A.5a.2a and 2a.3a.1, respectively. The isolated strains belonged to the same clade as the strains prevalent in Guangdong during the same period. No drug-resistant variations were detected in N1 or N2 gene, and the isolated strains were sensitive to oseltamivir and zanamivir.Conclusions:H1pdm subtype had stronger replication ability than H3 subtype in the influenza viruses isolated from co-infected cases. H1N1pdm and H3N2 subtype influenza viruses were genetically similar to the strains circulating in Guangdong at the same time. The isolated H1N1pdm and H3N2 strains were sensitive to both oseltamivir and zanamivir, indicating that they could continue to be used in the treatment of influenza virus infections caused by one or two genotypes.

Acute pancreatitis-associated ascites fluid (PAAF) is a common complication in patients with acute pancreatitis (AP) and is closely associated with the severity of AP, the development of local and systemic complications, and prognosis. PAAF may originate from the leakage of abdominal blood vessels, lymphatic vessels, and pancreatic duct. Recent studies have found that early removal of PAAF by abdominal paracentesis drainage can help to reduce systemic inflammation and alleviate pancreatitis-associated organ injury, thereby improving the conditions of patients with severe AP and reducing mortality. However, it is still not completely clear how PAAF aggravates systemic inflammatory response, participates in pancreatic injury and damage of distal organs, and leads to the aggravation of disease conditions in patients with AP. Therefore, this article gives an overview of PAAF and summarizes related studies in recent years, so as to provide directions for exploring the pathophysiological process and treatment of AP.