ObjectiveTo explore the impact of meteorological factors on influenza-like illness （ILI） cases in Anhui Province. MethodsILI surveillance data of Anhui Province from 2016 to 2023 were collected from the “China Influenza Surveillance Information System”， and meteorological data were obtained from Anhui Meteorological Center. R 4.2.1 was used to conduct Spearman rank correlation analysis between ILI cases and meteorological factors. A generalized additive model （GAM） was established to analyze the impact of meteorological factors on ILI cases. ResultsCorrelation analysis and GAM analysis showed that the average temperature and average relative humidity with a one-week lag had significant non-linear effects on ILI （P<0.05）. When the weekly average temperature was below 14 ℃， the number of ILI cases gradually increased as the temperature decreased. When the weekly average relative humidity was below 65%， the number of ILI cases gradually increased as the weekly average relative humidity decreased. The changes in ILI among the age groups 5-14， 15-24 and 25-59 years were influenced by meteorological factors， exhibiting a nonlinear relationship with weekly average temperature or weekly average relative humidity. ConclusionWeekly average temperature and weekly average relative humidity are the main factors affecting ILI in Anhui Province， and there is a lag effect， which can provide a reference for influenza surveillance and early warning.

ObjectiveTo investigate the trends in the disease burden of falls among Chinese children and adolescents from 1990 to 2021, and to inform the development of targeted prevention and control strategies. MethodsData on age-standardized incidence rate, age-standardized mortality rate and age-standardized disability-adjusted life years (DALYs) rate for falls among Chinese children and adolescents under 20 years of age from 1990 to 2021 were systematically extracted from the Global Burden of Disease Study (GBD) 2021 database. The annual percent change (APC) and average annual percent change (AAPC) were calculated to evaluate temporal trends in fall-related disease burden across different age groups and sexes. ResultsFrom 1990 to 2021, the age-standardized incidence rate of falls among Chinese children and adolescents showed a non-linear fluctuation (AAPC=0.08%, P=0.040), while both the age-standardized mortality rate and age-standardized DALYs rate declined (AAPC=-3.66% and -3.09%, both P<0.001). The crude incidence rate decreased in the <5 years old and 5 ‒ <10 years old groups (AAPC=-1.38% and -0.29%, both P<0.001), but increased annually in the 10 ‒ <15 years old and 15 ‒ <20 years old groups (AAPC=0.27% and 0.81%, both P<0.001). The crude mortality rate and crude DALYs rate declined across all four age groups, with the most significant reduction observed in the <5 years old group (AAPC=-5.21%, 95%CI:-5.55%‒ -5.00%; AAPC=-4.98%, 95%CI: -5.20%‒ -4.79%, both P<0.001). For both males and females, the age-standardized incidence rate of falls showed a V-shaped trend (initial decline followed by a subsequent rise). In contrast, the age-standardized mortality rate and age-standardized DALY rate declined in both sexes, with an AAPC of -3.40% and -2.94% for males, and -4.50% and -3.49% for females, respectively (all P<0.001). ConclusionThe overall disease burden of falls among Chinese children and adolescents has been substantially improved; however, the incidence burden remains considerable. Future prevention efforts should be tailored to age- and sex-specific characteristics, with the family as the fundamental care unit in conjunction with high-quality nursing extension services, to establish a closed-loop management model encompassing “early identification, immediate intervention, and continuous follow-up.”

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Objective The chemical composition and blood components of Tianmaxingnao capsules were discovered and examined using UPLC-Q/TOF-MS,and the possible pharmacological substance basis was preliminarily elucidated.Methods An UPLC-Q/TOF-MS method was developed in this study to determine the chemical composition of Tianma Xingnao capsules.After administration of Tianmaxingnao capsules,gather and examine rat plasma samples to investigate the exposed components of Tianmaxingnao capsules in rats.Results A total of 195 chemical components were identified in Tianmaxingnao capsules,including phenols,triterpenoid saponins,phenylethanol glycosides,cyclic ether terpenes,lipids,and phenylpropanoids.These components include those that are typical of Gastrodia elata Bl,Pheretima,Cistanche deserticola Ma,Rehmannia glutinosa,Polygala hybrida DC,and Acorus tatarinowii.Rat plasma samples were used to identify 37 prototype components and 3 metabolites of Tianmaxingnao capsules after they were administered.Conclusion This approach is easy to use,effective,sensitive,and precise.It may be used to investigate Tianmaxingnao capsules and the components that reach the bloodstream in detail,as well as to provide a first understanding of the pharmacological basis of the capsules.This study serves as a foundation for clarifying the pharmacological basis of Tianmaxingnao capsules and holds some reference value in exposing the pharmacological mechanism of the product.

Objective:To investigate the incidence rate and gene variation of methylmalonic academia (MMA) in Ji′nan city by analyzing biochemical and genetic screening results, and to explore the carrier frequency of MMA-related pathogenic genes in the population in Ji′nan.Methods:The children diagnosed with MMA by tandem mass spectrometry screening in Ji′nan Neonatal Disease Screening Centre from May 2011 to May 2022 were enrolled in this study.Their genetic test results were retrospectively analyzed and summarized.The dried heel blood tablets collected from 6 800 newborns were tested for neonatal gene screening. MMAA, MMAB, MMACHC and MMUT genes in 4 800 cases were detected by high-throughput sequencing+ target area capture technology.Ultra-multiplex polymerase chain reaction+ target gene locus capture technology was used to detect 174 target loci of 8 genes related to MMA in 2 000 cases.The hotspot mutation and related gene carrier rate of MMA were analyzed. Results:A total of 367 452 newborns were screened by tandem mass spectrometry, and 103 cases (56 males and 47 females) were diagnosed with MMA by screening.The estimated incidence of MMA was 1∶3 567.Among the 103 MMA cases, 76 were genetically diagnosed, and 4 gene variants of MMA ( MMAHC, MMUT, MMAA, MMADHC) were identified.A total of 6 800 neonates underwent neonatal genetic screening.Three of them were diagnosed with MMA.About 318 infants carried pathogenic variants of MMA, with a total carrier rate of 4.68%.Specifically, the carrier rates of MMACHC and MMUT gene variants were 3.09%(210/6 800) and 1.43% (97/6 800), respectively. Conclusions:MMA is the most common organic acid metabolism disorder in our country.The incidence and carrier rate of this disease are high in Jinan city.Neonatal genetic screening is an important supplement to neonatal biochemical screening.Carrier screening for MMA-related pathogenic genes is recommended for couples of childbearing age in Jinan.

Objective : To analyze the epidemiological characteristics and pathogen spectrum of hand，foot mouth disease (HFMD) in Anhui province from 2015 to 2022，and to provide scientific evidence for prevention and control measures of HFMD． Methods : The surveillance data of hand，foot and mouth disease in Anhui province from 2015 to 2022 were analyzed by descriptive epidemiology. Real-time PCR was used to detect and classify HFMD samples. Results : A total of 650 590 HFMD cases were reported in Anhui province from 2015 to 2022，including 1 406 se- vere cases and 17 deaths．The annual reported incidence was 131. 45 /100 000．The epidemic features of“low incidence in odd years and high incidence in even years”were presented from 2015 to 2019．The incidence showed a continuous decline from 2020 to 2022．The monthly distribution showed the characteristics of bimodal epidemic，and the main peak was not obvious in 2020．Hefei，Fuyang，Bozhou，Chuzhou and Suzhou ranked the top five cities in terms of cumulative incidence．The age of onset was mainly distributed in children aged 5 years and below，accounting for 89. 26% of the total cases．The male to female ratio was 1. 48 ∶ 1．A total of 28 657 laboratory-confirmed cases had been reported from 2015 to 2022．EV71 cases accounted for 10. 57% ，Cox A16 cases accounted for 24. 90% ，and other enterovirus cases accounted for 64. 53%．The dominant pathogens showed dynamic changes in different years．Since 2018，the proportion of EV71 decreased significantly，and the proportion of other enteroviruses gradually increased to become the dominant pathogens．Among other enteroviruses，Cox A6 strain was dominant (80. 48% ) ． Conclusion This study suggests that the prevention and control of HFMD in Anhui province should be paid more attention from April to July and from October to December．The focus areas are the cities in northern Anhui and Hefei where the floating population is large．The focus of prevention and control is on children aged 5 years and below．Other enteroviruses have become the dominant pathogens of hand-foot-mouth disease in Anhui province，Cox A6 strain is dominant.

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.
Animals ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; Humans ; Porcine Reproductive and Respiratory Syndrome/prevention & control* ; Porcine respiratory and reproductive syndrome virus/genetics* ; Swine

Antigenic purity is important for quality control of the foot-and-mouth (FMD) whole virus inactivated vaccine. The recommended method for evaluation the antigenic purity of FMD vaccine is to check the serum conversion to non-structural protein (NSP) 3AB antibody after 2 to 3 times inoculation of animals with inactivated vaccine. In this study, we developed a quantitative ELISA to detect the amount of residual 3AB in vaccine antigen, to provide a reference to evaluate the antigenic purity of FMD vaccine. Monoclonal antibody (Mab) of NSP 3A and HRP-conjugated Mab of NSP 3B were used to establish a sandwich ELISA to quantify the NSP 3AB in vaccine antigen of FMD. Purified NSP 3AB expressed in Escherichia coli was serially diluted and detected to draw the standard curve. The detectable limit was determined to be the lowest concentration of standard where the ratio of its OD value to OD blank well was not less than 2.0. Results: The OD value was linearly corelated with the concentration of 3AB protein within the range between 4.7 and 600 ng/mL. The correlation coefficient R² is greater than 0.99, and the lowest detectable limit is 4.7 ng/mL. The amount of 3AB protein in non-purified inactivated virus antigen was detected between 9.3 and 200 ng/mL depending on the 12 different virus strains, whereas the amount of 3AB in purified virus antigen was below the lowest detectable limit. The amount of 3AB in 9 batches of commercial FMD vaccine antigens was between 9.0 and 74 ng/mL, whereas it was below the detectable limit in other 24 batches of commercial vaccine antigens. Conclusion: the sandwich ELISA established in this study is specific and sensitive to detect the content of 3AB protein in vaccine antigen of FMD, which will be a useful method for evaluation of the antigenic purity and quality control of FMD inactivated vaccine.
Animals ; Antibodies, Viral ; Enzyme-Linked Immunosorbent Assay ; Foot-and-Mouth Disease/prevention & control* ; Foot-and-Mouth Disease Virus ; Viral Nonstructural Proteins/genetics* ; Viral Vaccines

Objective: By detecting the expression of interleukin-13 (IL-13) and periostin in the airway of asthmatic patients, the pathological changes and pulmonary functions of airway tissues in asthmatic patients were evaluated, and the role of IL-13 and periostin airway remodeling in bronchial asthma was preliminarily explored. Methods: The bronchial tissues adjacent to tumor nest were obtained from 12 patients with lung cancer complicated with bronchial asthma (asthmatic group) and 12 lung cancer patients without bronchial asthma (non-asthmatic group) after lung cancer resection. Pulmonary function was measured for all subjects before surgery. Pathological changes of airway tissues and degree of airway remodeling were assessed by hematoxylin-eosin (H&E) staining, masson′s trichrome staining, and periodic acid-silver methenamine (PASM) staining of paraffin-embedded sections. The expression of IL-13 and periostin in bronchial tissues were evaluated by immunohistochemistry. Results: Values of the forced expiratory volume in 1 second of the predicted value (FEV1% pred) and FEV1/forced vital capacity (FEV1/FVC%) in asthmatic patients were significantly decreased compared with the non-asthmatic patients (P<0.05), indicating that lung function was impaired in asthmatic patients. There was more severe airway remodeling representing as thickening of basement membranes, collagen deposition, and increasing of goblet cells and fibroblasts in asthmatic patients than in non-asthmatic patients (all P<0.05). The expression of IL-13 and periostin were higher in asthmatic tissues than in non-asthmatic tissues (P<0.05). The immunohistochemical expression of IL-13 and periostin in bronchial tissues were positively correlated with the degree of airway remodeling in asthmatic patients, and the expression of IL-13 and periostin in bronchial tissues were positively correlated with each other. Conclusions The expression of IL-13 and periostin were increased in bronchial tissue in patients with asthma. They work together to promote the occurrence of airway remodeling, which eventually lead to a decline in lung function.

To investigate the relationship between glucocorticoid receptor (GRα, GRβ) expression and hormone sensitivity in kaposiform hemangioendothelioma (KHE) patients complicated by Kasabach-Merritt phenomenon (KMP).Methods: In this study, 25 cases of KHE with KMP (mean age 2.4±1.5 months), including 9 males and 16 females at Henan Provincial People′s Hospital between May 2013 and May 2016 were included. All patients underwent surgical resection after being treated with hormone for one week, and were divided into sensitive group (10 cases) and resistance group (15 cases) according to the efficacy evaluation criteria. Normal tissues collected from 15 patients received surgical excision of lipoma was performed as a control group.Immunohistrochemical SP method was adopted in detecting GRα and GRβ expression in all groups. The expression intensity and positive rate were analyzed. Statistical significance was determined using the Wilcoxon rank sum test for the group samples and the Kruskal-Wallis test for multiple samples. Values of P<0.05 were considered statistically significant. Results: ①There was no significant difference among the control group, the resistance group and the sensitive group in expression intensity of GRα (control group 4.20±1.01, resistance group 4.10±0.99, sensitive group 3.53±0.52, P=0.632). The number of GRα positive cells in the sensitive group(46.20±2.57)was higher than that in the resistance group (40.93±5.18, P=0.032). ②The expression intensity of GRβ in the resistance group(5.40±0.51)was significantly higher than that in the sensitive group(2.60±0.52)and the control group(2.87±0.64, P=0.000, 0.002); there was significant difference among the sensitive group(29.70±2.50), the control group(36.07±3.47)and the resistance group (47.27±5.59)in the number of GRβ positive cells (P=0.000). ③The ratio of GRα/Grβ expression intensity was significantly lower in the resistance group than in the sensitive group and the control group; The ratio of GRα/GRβ positive cells was gradually decreased in the sensitive group, control group and resistance group, and the difference was statistically significant (P=0.000). Conclusions In the tumor tissue of KHE children with KMP, the increase of GRβ expression negatively regulats GRα, leading to imbalance of the expression ratio of the two, which may be an important cause for GC resistance.