ObjectiveTo analyze the epidemiological and etiological characteristics of a cluster of human metapneumovirus (HMPV) infection in a kindergarten in Gongshu District of Hangzhou City in May 2024, and to provide reference for the prevention and control of similar outbreaks. MethodsAn on-site investigation was conducted using an epidemiological case investigation form. Throat swab specimens collected from cases were screened for 13 respiratory pathogens using real-time fluorescent polymerase chain reaction (PCR). For HMPV nucleic acid positive specimens, the F gene of HMPV was used as the target gene for amplification and sequencing. The sequencing results were then compared with sequences in GenBank database to determine the virus subtypes and perform phylogenetic analyses. ResultsThe outbreak occurred in a kindergarter junior class with a total of 28 preschoolers and 3 teachers and childcare workers. A total of 11 cases (10 preschoolers and 1 teacher) were identified, including 8 male cases and 3 female cases. Clinical manifestations included fever in all 11 cases (100.00%), cough in 8 cases (72.72%), catarrhal symptoms in 4 cases (36.36%), and headache in 3 cases (27.27%). All symptoms were mild, and no severe cases were observed. A total of 11 throat swab samples were collected. Real-time fluorescent PCR test results showed that 3 samples were positive for HMPV nucleic acid, 2 samples were positive for both HMPV and Streptococcus pneumoniae, and 1 sample was positive for both HMPV and rhinovirus. The sequences of the 6 HMPV nucleic acid positive specimens were amplified and analyzed using specific primers, and all were determined to be HMPV subtype A2b. The F gene fragment sequence showed the highest similarity to PV081665.1/Brazil/2024 (99.65%), and also exhibited high similarity to PP683455.1/Indonesia/2021 (99.48%), PV016275.1/Beijing/2024 (99.31%), and PV052230.1/USA/2024 (99.13%). ConclusionThis cluster of acute respiratory tract infection was caused by HMPV subtype A2b, with co-infection of rhinovirus and Streptococcus pneumoniae. The F gene fragment sequences of the HMPV in this outbreak were highly homologous to those of the A2b strains isolated from Brazil, Beijing, Indonesia, and the the United States.

Objective To investigate the effect of long non-coding RNA non-coding RNA-activated by DNA damage(LncRNA NORAD)on macrophage apoptosis induced by Mycobacterium tuberculosis infection via sponging microRNA-20a-5p(miR-20a-5p).Methods Healthy subjects(n=50)who came for health checkup,patients with active tuberculosis(n=50)and individuals with asymptomatic M.tuberculosis infection(n=50)were enrolled from Hebei Chest Hospital from March 2022 to April 2023.Venous blood samples were collected to prepare serum samples.The expression levels of LncRNA NORAD,miR-20a-5p,and inflammatory factors in the serum were measured.Human monocyte line THP-1 was induced to differentiate into macrophages and assigned into Control group,Model group,transfection of NORAD empty vector group(sh-NC group),transfection of sh-NORAD vector group(sh-NORAD group),co-transfection of sh-NORAD and miR-20a-5p inhibitor empty vector group(sh-NORAD+miR-20a-5p inhibitor NC group),co-transfection of sh-NORAD and miR-20a-5p inhibitor vector group(sh-NORAD+miR-20a-5p inhibitor group),transfection of miR-20a-5p empty vector group(miR-NC group),and transfection of miR-20a-5p vector group(miR-20a-5p mimics group).The expression levels of LncRNA NORAD and miR-20a-5p(qRT PCR method),cell proliferation ability(CCK-8 kit method),cell apoptosis(flow cytometry method),inflammatory factor levels(ELISA method),and protein expression levels of BCL2-Associated X(Bax),B-cell lymphoma-2(Bcl-2),and cleaved caspase 3 in cells were detected.The targeted relationship between LncRNA NORAD and miR-20a-5p was validated.Results Compared with healthy subjects,the patients with active tuberculosis and asymptomatic M.tuberculosis infection had significantly higher serum levels of inflammatory factors and expression of LncRNA NORAD,and significantly lower miR-20a-5p.Compared with Control group,Model group had significantly higher LncRNA NORAD level,cell proliferation ability,Bcl-2 protein expression,and inflammatory factor levels,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NC group,the sh-NORAD group had significantly lower LncRNA NORAD level,Bcl-2 protein expression,inflammatory factor levels,and cell proliferation ability,but significantly higher miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NORAD+miR-20a-5p inhibitor NC group,the sh-NORAD+miR-20a-5p inhibitor group had significantly higher inflammatory factor levels,Bcl-2 protein expression,and cell proliferation ability,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the miR-NC group,the miR-20a-5p mimics group had significantly increased inflammatory cytokines and proliferation ability,and significantly reduced apoptosis rate(P＜0.05).The targeted relationship between LncRNA NORAD and miR-20a-5p was further confirmed through experiments.Conclusions LncRNA NORAD is overexpressed in macrophages induced by M.tuberculosis.Silencing the expression of LncRNA NORAD can target the downregulation of miR-20a-5p expression,thereby inhibiting the inflammatory response of macrophages induced by M.tuberculosis and promoting cell apoptosis.

Objective To investigate the effect of long non-coding RNA non-coding RNA-activated by DNA damage(LncRNA NORAD)on macrophage apoptosis induced by Mycobacterium tuberculosis infection via sponging microRNA-20a-5p(miR-20a-5p).Methods Healthy subjects(n=50)who came for health checkup,patients with active tuberculosis(n=50)and individuals with asymptomatic M.tuberculosis infection(n=50)were enrolled from Hebei Chest Hospital from March 2022 to April 2023.Venous blood samples were collected to prepare serum samples.The expression levels of LncRNA NORAD,miR-20a-5p,and inflammatory factors in the serum were measured.Human monocyte line THP-1 was induced to differentiate into macrophages and assigned into Control group,Model group,transfection of NORAD empty vector group(sh-NC group),transfection of sh-NORAD vector group(sh-NORAD group),co-transfection of sh-NORAD and miR-20a-5p inhibitor empty vector group(sh-NORAD+miR-20a-5p inhibitor NC group),co-transfection of sh-NORAD and miR-20a-5p inhibitor vector group(sh-NORAD+miR-20a-5p inhibitor group),transfection of miR-20a-5p empty vector group(miR-NC group),and transfection of miR-20a-5p vector group(miR-20a-5p mimics group).The expression levels of LncRNA NORAD and miR-20a-5p(qRT PCR method),cell proliferation ability(CCK-8 kit method),cell apoptosis(flow cytometry method),inflammatory factor levels(ELISA method),and protein expression levels of BCL2-Associated X(Bax),B-cell lymphoma-2(Bcl-2),and cleaved caspase 3 in cells were detected.The targeted relationship between LncRNA NORAD and miR-20a-5p was validated.Results Compared with healthy subjects,the patients with active tuberculosis and asymptomatic M.tuberculosis infection had significantly higher serum levels of inflammatory factors and expression of LncRNA NORAD,and significantly lower miR-20a-5p.Compared with Control group,Model group had significantly higher LncRNA NORAD level,cell proliferation ability,Bcl-2 protein expression,and inflammatory factor levels,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NC group,the sh-NORAD group had significantly lower LncRNA NORAD level,Bcl-2 protein expression,inflammatory factor levels,and cell proliferation ability,but significantly higher miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NORAD+miR-20a-5p inhibitor NC group,the sh-NORAD+miR-20a-5p inhibitor group had significantly higher inflammatory factor levels,Bcl-2 protein expression,and cell proliferation ability,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the miR-NC group,the miR-20a-5p mimics group had significantly increased inflammatory cytokines and proliferation ability,and significantly reduced apoptosis rate(P＜0.05).The targeted relationship between LncRNA NORAD and miR-20a-5p was further confirmed through experiments.Conclusions LncRNA NORAD is overexpressed in macrophages induced by M.tuberculosis.Silencing the expression of LncRNA NORAD can target the downregulation of miR-20a-5p expression,thereby inhibiting the inflammatory response of macrophages induced by M.tuberculosis and promoting cell apoptosis.

OBJECTIVE To identify Bupleurum chinense pieces and its counterfeits based on the characteristic chromatogram and UPLC-Q-TOF/MS technology.METHODS Thin layer chromatography was used to identify Bupleurum chinense and its counterfeits collected from different origins and different producing areas.Then,the chemical constituents of Bupleurum chinense and its counter-feits were compared according to the established HPLC characteristic chromatogram,and the representative differential markers of Bup-leurum chinense counterfeits were further identified by UPLC-Q-TOF/MS technology.RESULTS Thin layer chromatography showed that different original Bupleurum chinense pieces had saikosaponin A and saikosaponin D,which could be distinguished according to the intensity and position of fluorescent spots.There were 14 and 16 common peaks in the specific chromatogram of Bupleurum chinense and its vinegar-processed products respectively,and 6 components were identified.The chemical components of Bupleurum chinense pieces from different producing areas were similar,and the established method could better reflect the characteristics of Bupleurum chinense.Further comparison of the specific chromatogram of Bupleurum chinense and its counterfeits showed that the composition of B.marginatum Wall.was close to that of the authentic Bupleurum chinense.,B.bicaule Helm and B.longiradiatum Turcz.had their own characteristic peaks with high response values.The content of saponins in B.marginatum var.stenophyllum was significantly high-er.A total of 69 Bupleurum compounds were identified by UPLC-Q-TOF/MS mainly triterpenoid saponins,followed by flavonoids,a few chromones,phenylpropanoids and alkynes.The results of cluster analysis showed that the interspecific differences of Bupleurum chinense were obvious,and different Bupleurum counterfeits had representative differential markers.CONCLUSION The established characteristic chromatogram and UPLC-Q-TOF/MS method can be used for the chemical identification of Bupleurum chinense and its counterfeits,which provide a basis for the comprehensive and rational development and utilization of Bupleurum resources.

BACKGROUND:In recent years,with the deepening of cell death research,necroptosis has gradually become a hot and difficult topic in academic research.Its various signaling pathways and proteins play an important role in the occurrence and development of periodontitis. People try to delay the process of periodontal tissue inflammation by preventing the occurrence of necroptosis. OBJECTIVE:To provide a review on the molecular mechanism of necroptosis and its correlation with periodontitis in the hope of providing new ideas and directions for the prevention,diagnosis,treatment and evaluation of periodontitis. METHODS:The first author searched PubMed,CNKI,and other databases in October 2023 for relevant literature published up to April 2024 with the search terms of "necroptosis,programmed cell death,periodontitis,periodontal,immunity,inflammation,receptor interacting protein kinase 1,receptor interacting protein kinase 3,mixed lineage kinase domain-like" in Chinese and "necroptosis,programmed cell death,periodontitis,periodontal,immunity,inflammation,RIP1,RIP3,MLKL" in English,respectively. The titles and abstracts of each document were read for preliminary screening,and 56 documents were finally selected for generalization and analysis. RESULTS AND CONCLUSION:(1) The main pathogenesis of periodontitis is that periodontal pathogenic bacteria stimulate the organism,leading to changes in the periodontal microenvironment,breaking the original immune balance and releasing a variety of inflammatory factors,triggering an inflammatory response resulting in destruction of periodontal tissues. Necroptosis can modulate the immuno-inflammation in periodontal tissues,thus affecting the development of periodontitis. (2) The occurrence of necroptosis is related to a variety of proteins,signaling pathways,and signaling molecules. Animal experiments have confirmed that the expression of phosphorylated mixed lineage kinase domain-like protein (pMLKL) is related to the degree of inflammation,and RIP3 and pMLKL are highly expressed in the periodontal tissues of mice suffering from periodontitis. Blocking the RIP3/MLKL pathway is conducive to the reduction of inflammation in periodontal tissues and the control of periodontitis progression. (3) To explore whether pMLKL can be used as a diagnostic indicator of periodontitis is of great value in the prevention,diagnosis and treatment of periodontitis.

Objective:To explore the current status and factors influencing physical activity among pre-frail and frail older adults in the community.Methods:Convenience sampling was used to select 207 pre-frail and frail older adults from Donghuashi community and Fangzhuang community in Beijing from April to June 2024 as study subjects. Older adults were surveyed for their general information, lifestyle behaviors, nutritional status, and physical activity. Binary Logistic regression was used to explore the factors influencing the level of physical activity among pre-frail and frail older adults in the community.Results:A total of 207 questionnaires were distributed and 204 valid questionnaires were recovered, with a valid recovery rate of 98.55%. The Physical Activity Scale for the Elderly score of the 204 community-based pre-frail and frail older adults was 86.87 (52.14, 125.00), and the form of activity was predominantly walking (98.5%, 201/204) and light domestic physical activity (85.8%, 175/204). Binary Logistic regression showed that taking a nap ( OR=3.614), abnormal nighttime sleep duration ( OR=4.077), fear of falling before or during exercise ( OR=7.895), and risk of malnutrition ( OR=9.263) were risk factors for levels of physical activity in pre-frail and frail older adults in the community ( P<0.05), and good exercise cognition ( OR=0.055) was a protective factor for physical activity levels ( P<0.05) . Conclusions:Pre-frail and frail older adults in the community have low levels of physical activity, which is dominated by walking and household activities. Community healthcare workers should strengthen the management of physical activity for pre-frail and frail older adults, cultivate their good living habits, improve their sleep quality, ensure sufficient night sleep, help them overcome the fear of falling before or during exercise, set up the correct concept of exercise, form a good cognition of exercise, and guide their family members to pay attention to the nutritional status of older adults, beware of the risk of malnutrition, and improve the level of physical activity to delay or even reverse the frail state.

Objective To assess the potential role of disulfidptosis-related genes (DRGs) in pan-cancer on prognosis and immunity on the basis of bioinformatics approaches. Methods Pan-cancer RNA-seq data, mutation profiles, clinical information, TMB, MSI, stemness scores, and tumor and immune microenvironment data contained in TCGA and various open-source online databases, and multi-group R-language algorithms were used for comprehensive analysis. The expression levels of DRGs at the cellular level were experimentally validated using qPCR. Results LRPPRC, NCKAP1, NDUFS1, and NUBPL had a better prognosis in renal clear cell carcinoma (P<0.001), whereas SLC7A11, NCKAP1, and SLC3A2 had a worse prognosis in hepatocellular carcinoma (P<0.001). TME analysis showed that LRPPRC was negatively correlated with immune cells, stromal cells, and estimated scores in all tumor types. TMB analysis revealed the potential research value of DRGs for PD-1/PD-L1 therapy in pan-cancer. Drug sensitivity analysis showed that SLC7A11 (r=0.454), SLC3A2 (r=0.366), and NCKAP1 (r=0.455) were significantly associated with Kahalide F (P<0.01). Experimental validation demonstrated the overall higher expression levels of GYS1 and NCKAP1 than normal cells in lung adenocarcinoma, colon adenocarcinoma, esophageal squamous carcinoma, and hepatocellular carcinoma (P<0.05). Conclusion Pan-cancer analysis of DRGs indicates that DRGs may serve as important biomarkers for the diagnosis and prognosis of renal clear-cell carcinoma, lung adenocarcinoma, and hepatocellular carcinoma.

OBJECTIVE To identify Bupleurum chinense pieces and its counterfeits based on the characteristic chromatogram and UPLC-Q-TOF/MS technology.METHODS Thin layer chromatography was used to identify Bupleurum chinense and its counterfeits collected from different origins and different producing areas.Then,the chemical constituents of Bupleurum chinense and its counter-feits were compared according to the established HPLC characteristic chromatogram,and the representative differential markers of Bup-leurum chinense counterfeits were further identified by UPLC-Q-TOF/MS technology.RESULTS Thin layer chromatography showed that different original Bupleurum chinense pieces had saikosaponin A and saikosaponin D,which could be distinguished according to the intensity and position of fluorescent spots.There were 14 and 16 common peaks in the specific chromatogram of Bupleurum chinense and its vinegar-processed products respectively,and 6 components were identified.The chemical components of Bupleurum chinense pieces from different producing areas were similar,and the established method could better reflect the characteristics of Bupleurum chinense.Further comparison of the specific chromatogram of Bupleurum chinense and its counterfeits showed that the composition of B.marginatum Wall.was close to that of the authentic Bupleurum chinense.,B.bicaule Helm and B.longiradiatum Turcz.had their own characteristic peaks with high response values.The content of saponins in B.marginatum var.stenophyllum was significantly high-er.A total of 69 Bupleurum compounds were identified by UPLC-Q-TOF/MS mainly triterpenoid saponins,followed by flavonoids,a few chromones,phenylpropanoids and alkynes.The results of cluster analysis showed that the interspecific differences of Bupleurum chinense were obvious,and different Bupleurum counterfeits had representative differential markers.CONCLUSION The established characteristic chromatogram and UPLC-Q-TOF/MS method can be used for the chemical identification of Bupleurum chinense and its counterfeits,which provide a basis for the comprehensive and rational development and utilization of Bupleurum resources.

BACKGROUND:In recent years,with the deepening of cell death research,necroptosis has gradually become a hot and difficult topic in academic research.Its various signaling pathways and proteins play an important role in the occurrence and development of periodontitis. People try to delay the process of periodontal tissue inflammation by preventing the occurrence of necroptosis. OBJECTIVE:To provide a review on the molecular mechanism of necroptosis and its correlation with periodontitis in the hope of providing new ideas and directions for the prevention,diagnosis,treatment and evaluation of periodontitis. METHODS:The first author searched PubMed,CNKI,and other databases in October 2023 for relevant literature published up to April 2024 with the search terms of "necroptosis,programmed cell death,periodontitis,periodontal,immunity,inflammation,receptor interacting protein kinase 1,receptor interacting protein kinase 3,mixed lineage kinase domain-like" in Chinese and "necroptosis,programmed cell death,periodontitis,periodontal,immunity,inflammation,RIP1,RIP3,MLKL" in English,respectively. The titles and abstracts of each document were read for preliminary screening,and 56 documents were finally selected for generalization and analysis. RESULTS AND CONCLUSION:(1) The main pathogenesis of periodontitis is that periodontal pathogenic bacteria stimulate the organism,leading to changes in the periodontal microenvironment,breaking the original immune balance and releasing a variety of inflammatory factors,triggering an inflammatory response resulting in destruction of periodontal tissues. Necroptosis can modulate the immuno-inflammation in periodontal tissues,thus affecting the development of periodontitis. (2) The occurrence of necroptosis is related to a variety of proteins,signaling pathways,and signaling molecules. Animal experiments have confirmed that the expression of phosphorylated mixed lineage kinase domain-like protein (pMLKL) is related to the degree of inflammation,and RIP3 and pMLKL are highly expressed in the periodontal tissues of mice suffering from periodontitis. Blocking the RIP3/MLKL pathway is conducive to the reduction of inflammation in periodontal tissues and the control of periodontitis progression. (3) To explore whether pMLKL can be used as a diagnostic indicator of periodontitis is of great value in the prevention,diagnosis and treatment of periodontitis.

Objective:To discuss the promotive effect of Shengji Yuhong Ointment extract(SYOE)on skin wound healing of zebrafish,and to clarify its mechanism.Methods:A total of 320 wild-type AB strain zebrafish were used to establish a wound model by making an incision on the lateral abdomen.The zebrafish were randomly divided into control group,low dose of SYOE group(raised in water containing 0.625 mg·L-1 SYOE),high dose of SYOE group(raised in water containing 1.250 mg·L-1 SYOE),and allantoin group(raised in water containing 1.000 mng·L-1 allantoin),and there were 80 zebrafish in each group.The wound areas were recorded by photographs on days 7,14,and 21 after injury,and the wound healing rates were calculated.The skin tissue samples from the wound sites of the zebrafish in various groups were collected at different time points to prepare histological sections.HE staining was used to observe the widths of skin wound edges of the zebrafish on days 0,7,14,and 21 in various groups;Sirius red staining was used to detect the collagen levels in the wound tissues of the zebrafish on days 2,4,6,and 8 after injury in various groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of type Ⅰ collagen(Col Ⅰ)and α-smooth muscle actin(α-SMA)in the wound tissues on day 4 after injury;real-time quantitative PCR(RT-qPCR)was used to analyze the mRNA expression levels of Col Ⅰ-encoding genes(col1a1a,col1a1b,and col1a2)and α-SMA as wall as key factors of the transforming growth factor β(TGF-β)/Smad signaling pathway(tgfb1a,smad2,and smad3a)in the wound tissues of the zebrafish in various groups on day 4 after injury.Results:The wound healing assessment results showed that compared with control group,the wound healing rates of the zebrafish in low dose of SYOE group,high dose of SYOE group,and allantoin group were significantly increased on days 7,14,and 21 after injury(P＜0.05 or P＜0.01);the wound healing rate in high dose of SYOE group achieved 84％on day 21.The HE staining results showed that compared with control group,the widths of skin wound edges of the zebrafish in low dose of SYOE group,high dose of SYOE group,and allantoin group were significantly decreased on days 14 and 21 after injury(P＜0.05 or P＜0.01).The Sirius red staining results showed that compared with control group,the collagen levels in the skin wound tissue of the zebrafish in low dose of SYOE group,high dose of SYOE group,and allantoin group were significantly increased on days 6 and 8 after injury(P＜0.01).The ELISA results showed that on day 4 after injury,compared with control group,the levels of Col Ⅰ and α-SMA in the skin wound tissue of the zebrafish in low dose of SYOE group,high dose of SYOE group,and allantoin group were significantly increased(P＜0.05 or P＜0.01).The RT-qPCR results showed that on day 4 after injury,compared with control group,the expression levels of col1a1b,col1a2,α-SMA,tgfb1a,and smad2 mRNA in the skin wound tissue of the zebrafish in low dose of SYOE group were significantly increased(P＜0.05 or P＜0.01),while the expression levels of col1a1a,col1a1b,col1a2,α-SMA,tgfb1a,smad2,and smad3a mRNA in the skin wound tissue of the zebrafish in high dose of SYOE group and allantoin group were significantly increased(P＜0.01).Conclusion:SYOE can increase the collagen deposition in skin wound of zebrafish,promote wound healing,and upregulate the expression of genes related to the TGF-β/Smad signaling pathway.