Objective:To obtain fucose-free anti-West Nile virus nonstructural protein 1 (NS1) antibody and evaluate its antibody-dependent cell-mediated cytotoxicity (ADCC).Methods:The guanosine diphosphate-fucose transporter SLC35C1 in CHO cells was knocked out using CRISPR/Cas9 gene editing technology to obtain the fucose-free cell line CHO SLC35C1 -/-. CHO SLC35C1 -/- cells were used to produce fucose-free anti-West Nile virus NS1 antibodies. The binding abilities of the antibodies to the target antigen of West Nile virus NS1 protein and the human high-affinity IgG Fc receptor hFcγRⅠ (hCD64) were detected by ELISA and flow cytometry, respectively. The ADCC activity of the antibodies was detected by ADCC reporter gene assay. One-way analysis of variance was used for statistical analysis. Results:CHO SLC35C1 -/- cells expressed green fluorescent protein but not Lens culinaris agglutinin. The anti-West Nile virus NS1 antibodies produced by CHO SLC35C1 -/- cells with a fucose content of 0.22% could bind to West Nile virus NS1 protein in a concentration-dependent manner. Compared with the wild-type antibodies, the fucose-free anti-West Nile virus NS1 antibodies showed a stronger binding ability to hFcγRⅠ(hCD64), as indicated by a significant increase in fluorescence intensity. The ADCC reporter gene assay showed that the fucose-free anti-West Nile virus NS1 antibodies had increased activity as compared with the wild-type antibodies ( P<0.001). Conclusion:The fucose-free anti-West Nile virus NS1 antibodies may be used to protect against West Nile virus infection.

Objective:To obtain fucose-free anti-West Nile virus nonstructural protein 1 (NS1) antibody and evaluate its antibody-dependent cell-mediated cytotoxicity (ADCC).Methods:The guanosine diphosphate-fucose transporter SLC35C1 in CHO cells was knocked out using CRISPR/Cas9 gene editing technology to obtain the fucose-free cell line CHO SLC35C1 -/-. CHO SLC35C1 -/- cells were used to produce fucose-free anti-West Nile virus NS1 antibodies. The binding abilities of the antibodies to the target antigen of West Nile virus NS1 protein and the human high-affinity IgG Fc receptor hFcγRⅠ (hCD64) were detected by ELISA and flow cytometry, respectively. The ADCC activity of the antibodies was detected by ADCC reporter gene assay. One-way analysis of variance was used for statistical analysis. Results:CHO SLC35C1 -/- cells expressed green fluorescent protein but not Lens culinaris agglutinin. The anti-West Nile virus NS1 antibodies produced by CHO SLC35C1 -/- cells with a fucose content of 0.22% could bind to West Nile virus NS1 protein in a concentration-dependent manner. Compared with the wild-type antibodies, the fucose-free anti-West Nile virus NS1 antibodies showed a stronger binding ability to hFcγRⅠ(hCD64), as indicated by a significant increase in fluorescence intensity. The ADCC reporter gene assay showed that the fucose-free anti-West Nile virus NS1 antibodies had increased activity as compared with the wild-type antibodies ( P<0.001). Conclusion:The fucose-free anti-West Nile virus NS1 antibodies may be used to protect against West Nile virus infection.

A new method was developed for simultaneous and efficient determination of linear perfluorooctanoic acid(n-PFOA)and linear perfluorooctane sulfonate(n-PFOS),and their typical branched isomers in seawater by online solid phase extraction-liquid chromatography-tandem mass spectrometry(Online SPE-LC-MS/MS).Only centrifugation of the seawater sample was required to remove the particulate matter,and then the seawater sample was directly injected and analyzed by online SPE-LC-MS/MS.An Eclipse Plus-C18 guard column was selected as SPE column for online enrichment of linear and branched isomers,and a F5 PFP column(150 mm×2.1 mm,2.7 μm)was used as the analytical column.Under the optimized experimental conditions,the separation and detection of all PFOA and PFOS linear and branched isomers could be completed within 20 min.The spiked recoveries of various target compounds ranged from 82.9%to 107.7%with detection limits and limits of quantification of 0.10-1.05 ng/L and 0.30-2.11 ng/L,respectively.The method was characterized by good precision(RSD≤9.10%)and linearity(R2≥0.990).Subsequently,linear and branched isomers of PFOA and PFOS in surface and bottom seawater samples collected from the Laizhou Bay of China were determined.The results showed that the detection rate of all the four branched PFOA isomers were 100%,with the highest average concentration of 25.85 ng/L found for 6m-PFOA,which accounted for 11.79%of the∑PFOA.For the five branched isomers of PFOS,the highest detection rate of 90.84%was found for 5m-PFOS.The highest average concentration of 0.64 ng/L was observed for 3m-PFOS,accounting for 19.88%of ∑PFOS.The proposed method provided an effective detection tool for qualitative and quantitative detection of PFOA and PFOS isomers in the marine aquatic environment.

Skeletal muscle is the largest metabolic and endocrine organ in the body.As the basic unit of skeletal muscle,muscle fiber has high plasticity.Skeletal muscle fibers are mainly divided into oxidative and glycolytic muscle fibers,and muscle fibers type is an important factor affecting the contraction and energy metabolism of skeletal muscle.Understanding the molecular mechanism regulating skeletal muscle fiber type conversion is of great significance for regulating skeletal muscle-related diseases.Mitochondria is the energy factory of cell life activities,and the characteristics of mitochondria,namely the content,shape and distribution of mitochondria,are closely related to the function of mitochondria.The mitochon-drial characteristics of various types of muscle fiber are different,which is related to the difference of en-ergy metabolism of different muscle fiber types.Mitochondrial homeostasis is a dynamic equilibrium process,which is usually regulated by mitochondrial biogenesis,mitochondrial fusion and fission,mito-chondrial autophagy and other processes.These processes not only affect the morphology and quantity of mitochondria,but also affect the metabolic balance of glucose and fatty acids in the body.Many studies have also shown that mitochondria-mediated changes in the substrates of energy metabolism of skeletal muscle affect the type conversion process of skeletal muscle fiber.Exercise is a non-drug treatment that can generally promote the formation of oxidized muscle fibers by maintaining skeletal muscle mitochondri-al homeostasis.In this paper,the mitochondrial characteristics of different types of skeletal muscle fibers and the role of maintaining mitochondrial homeostasis in regulating muscle fiber type conversion were re-viewed.On this basis,the molecular mechanism of mitochondrial involvement in PGC1α,Ca2+and ROS signaling pathways mediated by fiber type conversion of skeletal muscle was summarized.Mitochondria are the energy factories of skeletal muscle,and targeted intervention through understanding their regulato-ry mechanism may be a new direction for the treatment of skeletal muscle-related diseases in the future.

OBJECTIVE: This study aimed to explore the association between body mass index (BMI) and mortality based on the 10-year population-based multicenter prospective study. METHODS: A general population-based multicenter prospective study was conducted at four sites in rural China between 2013 and 2023. Multivariate Cox proportional hazards models and restricted cubic spline analyses were used to assess the association between BMI and mortality. Stratified analyses were performed based on the individual characteristics of the participants. RESULTS: Overall, 19,107 participants with a sum of 163,095 person-years were included and 1,910 participants died. The underweight (< 18.5 kg/m ²) presented an increase in all-cause mortality (adjusted hazards ratio [ aHR] = 2.00, 95% confidence interval [ CI]: 1.66-2.41), while overweight (≥ 24.0 to < 28.0 kg/m ²) and obesity (≥ 28.0 kg/m ²) presented a decrease with an aHR of 0.61 (95% CI: 0.52-0.73) and 0.51 (95% CI: 0.37-0.70), respectively. Overweight ( aHR = 0.76, 95% CI: 0.67-0.86) and mild obesity ( aHR = 0.72, 95% CI: 0.59-0.87) had a positive impact on mortality in people older than 60 years. All-cause mortality decreased rapidly until reaching a BMI of 25.7 kg/m ² ( aHR = 0.95, 95% CI: 0.92-0.98) and increased slightly above that value, indicating a U-shaped association. The beneficial impact of being overweight on mortality was robust in most subgroups and sensitivity analyses. CONCLUSION This study provides additional evidence that overweight and mild obesity may be inversely related to the risk of death in individuals older than 60 years. Therefore, it is essential to consider age differences when formulating health and weight management strategies.
Humans ; Body Mass Index ; China/epidemiology* ; Male ; Female ; Middle Aged ; Prospective Studies ; Rural Population/statistics & numerical data* ; Aged ; Follow-Up Studies ; Adult ; Mortality ; Cause of Death ; Obesity/mortality* ; Overweight/mortality*

Skeletal muscle is the largest metabolic and endocrine organ in the body.As the basic unit of skeletal muscle,muscle fiber has high plasticity.Skeletal muscle fibers are mainly divided into oxidative and glycolytic muscle fibers,and muscle fibers type is an important factor affecting the contraction and energy metabolism of skeletal muscle.Understanding the molecular mechanism regulating skeletal muscle fiber type conversion is of great significance for regulating skeletal muscle-related diseases.Mitochondria is the energy factory of cell life activities,and the characteristics of mitochondria,namely the content,shape and distribution of mitochondria,are closely related to the function of mitochondria.The mitochon-drial characteristics of various types of muscle fiber are different,which is related to the difference of en-ergy metabolism of different muscle fiber types.Mitochondrial homeostasis is a dynamic equilibrium process,which is usually regulated by mitochondrial biogenesis,mitochondrial fusion and fission,mito-chondrial autophagy and other processes.These processes not only affect the morphology and quantity of mitochondria,but also affect the metabolic balance of glucose and fatty acids in the body.Many studies have also shown that mitochondria-mediated changes in the substrates of energy metabolism of skeletal muscle affect the type conversion process of skeletal muscle fiber.Exercise is a non-drug treatment that can generally promote the formation of oxidized muscle fibers by maintaining skeletal muscle mitochondri-al homeostasis.In this paper,the mitochondrial characteristics of different types of skeletal muscle fibers and the role of maintaining mitochondrial homeostasis in regulating muscle fiber type conversion were re-viewed.On this basis,the molecular mechanism of mitochondrial involvement in PGC1α,Ca2+and ROS signaling pathways mediated by fiber type conversion of skeletal muscle was summarized.Mitochondria are the energy factories of skeletal muscle,and targeted intervention through understanding their regulato-ry mechanism may be a new direction for the treatment of skeletal muscle-related diseases in the future.

Objective: To investigate the species composition, seasonal variation, and virus-carrying status of mosquitoes in the Honghe Wetland of Tengzhou City to better control local mosquito vectors. Methods: Seven surveillance sites were established in different scenic spots of the wetland, and ecological monitoring was carried out monthly from April to November each year using the light-trapping method. From June to September 2023, a total of 3 000 Mansonia uniformis mosquitoes were tested for flaviviruses using the nested PCR method. Results: From 2022 to 2024, a total of 153 mosquito trap lamp·nights were deployed in the Honghe Wetland, and 20 336 female mosquitoes were captured, with an average density of 132.92 mosquitoes/lamp·night. The mosquitoes were identified as belonging to 19 species across 6 genera. Among them, Lutzia chiangmaiensis was identified for the first time in mainland China, and similarly for Anopheles pullu, Culex inatomii, and Aedes togoi in southern Shandong Province. The dominant species was Ma. uniformis (15 345, 75.46%), followed by Culex tritaeniorhynchus (1824, 8.97%), Coquillettidia ochracea (1408, 6.92%), and Anopheles sinensis (1 026, 5.05%). The peak period of adult mosquito activity was from June to September. The seasonal variations in the density of the different mosquito species differed, with the peak periods of Cq. ochracea, An. sinensis, Ma. uniformis, and Cx. tritaeniorhynchus being in June, June, July, and August, respectively. The density of Cx. pipiens pallens was consistently low, showing an upward trend in October only. The 3 000 Ma. uniformis mosquitoes were negative for the three viruses tested. Conclusions The mosquitoes in the Honghe Wetland of Tengzhou City are rich in species composition and high in density. Because of a possible risk of transmission of Japanese encephalitis and malaria in the period between May and September, comprehensive mosquito control measures need to be strengthened in the Honghe Wetland of Tengzhou City.

Objective:To establish an in vivo infection model of West Nile virus (WNV) pseudovirus and evaluate the neutralizing activity of antibody WNV-XH1.Methods:A stable cell line that can package the WNV pseudovirus was established in the early stage to prepare the pseudovirus supernatant. The supernatant was concentrated and infected BHK21 cells to detect the titer of the pseudovirus. After intraperitoneal injection of the pseudovirus into C57BL/J mice, bioluminescence imaging was performed to observe the infection status of the pseudovirus in the mice. After simultaneous infection, blood was collected and ELISA was used to detect NS1 levels in mouse serum. The in vivo functional activity of antibody WNV-XH1 was evaluated using the established mouse infection model.Results:Fluorescence was detected in C57BL/J mice infected with WNV pseudovirus, and the NS1 levels in the peripheral blood serum of mice infected with pseudovirus were significantly higher than those of non infected mice (1.453±0.09vs0.305±0.018). After intravenous administration of WNV-XH1 antibody before the attack, the fluorescence signal in the mice decreased and the serum NS1 level decreased (0.384±0.015).Conclusions:A successful in vivo infection model of WNV pseudovirus was established, and it was confirmed that the antibody WNV-XH1 had a protective effect against WNV pseudovirus infection in vivo.

Osteoporotic proximal humeral fracture (OPHF) is one of the common osteoporotic fractures in the aged, with an incidence only lower than vertebral compression fracture, hip fracture, and distal radius fracture. OPHF, secondary to osteoporosis and characterized by poor bone quality, comminuted fracture pattern, slow healing, and severely impaired shoulder joint function, poses a big challenge to the current clinical diagnosis and treatment. In the field of diagnosis, treatment, and rehabilitation of OPHF, traditional Chinese and Western medicine have accumulated rich experience and evidence from evidence-based medicine and achieved favorable outcomes. However, there is still a lack of guidance from a relevant consensus as to how to integrate the advantages of the two medical systems and achieve the integrated diagnosis and treatment. To promote the diagnosis and treatment of OPHF with integrated traditional Chinese and Western medicine, relevant experts from Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine have been organized to formulate Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine ( version 2024) by searching related literatures and based on the evidences from evidence-based medicine. This consensus consists of 13 recommendations about the diagnosis, treatment and rehabilitation of OPHF with integrated traditional Chinese medicine and Western medicine, aimed at standardizing, systematizing, and personalizing the diagnosis and treatment of OPHF with integrated traditional Chinse and Western medicine to improve the patients ′ function.

A new method for simultaneous determination of 23 kinds of per-and polyfluoroalkyl substances(PFASs)(13 kinds of perfluoro carboxylic acids,4 kinds of perfluoro sulfonic acids,and 6 kinds of new substitutes)in plant leaf tissue by ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)using automatic online solid phase extraction(SPE)to remove the matrix interference components in plant crude extracts was developed.The plant leaf samples were extracted twice with 1%formic acid-methanol solution,then evaporated to dry,redissolved with 70%methanol solution,and directly injected for analysis.After 23 kinds of target PFASs were purified automatically by online SPE with a WAX column,the six-way valve was switched to rinse PFASs onto an alkaline mobile phase system-compatible C18 analytical column.Then,the 23 kinds of target PFASs were separated within 16 min by gradient elution using a binary mobile phase system of methanol/water(Containing 0.4%ammonium hydroxide).Tandem mass spectrometry was performed in multiple reaction monitoring(MRM)mode for online detection of various PFASs,and quantification was carried out by internal standard method.The results of the method validation showed that satisfactory average recoveries of 23 kinds of PFASs in plant leaf samples(64.2%-125.5%),precision(relative standard deviations(RSDs)of 0.7%-12.8%),linearity(R2＞0.990),and sensitivity(the detection limits(S/N=3)were in the range of 0.02-0.50 μg/kg)were achieved.Finally,this method was used to detect PFASs in the marine green tide algae(Enteromorpha prolifera)and several tree leaves,and a total of 6 kinds of PFASs were detected,in which PFBA was the main contaminant.Compared with the reported offline SPE methods,the proposed online SPE technique significantly simplified the sample pretreatment process and provided an automatic,simple,and environment-friendly method for the routine monitoring of legacy and emerging PFASs in plant tissues.