Objective To analyze the clinical efficacy of real-world bulleyaconitine A tablets combined with salt and pepper seven-seed hot package in the treatment of knee osteoarthritis(KOA).Methods 110 outpatient patients from the Department of Orthopedics,Shenzhen Traditional Chinese Medicine Hospital from December 2023 to August 2024 were selected,they were randomly divided into control group of 53 cases and treatment group of 57 cases.The patients in control group were treated with flurbiprofen gel paste,the patients in treatment group received oral administration of bulleyaconitine A tablets and external application of Jiaoyan Qizi hot package,the treatment course were all 4 weeks.All patients were followed up for 2 weeks after the end of treatment.Western Ontario and McMaster University osteoarthritis index(WOMAC)score,Lequesne score,12-item short form health survey(SF-12)score and clinical efficacy were compared between two groups,and adverse events were recorded.Results The total effective rate of treatment group was 87.7％,which was significantly higher than 71.7％of control group(P＜0.05);After treatment,WOMAC score,Lequesne score and SF-12 score in two groups were better than before treatment(P＜0.05).The improvement of WOMAC score,Lequesne score and SF-12 score in treatment group were more obvious(P＜0.05).Conclusion For KOA patients with cold and damp obstruction syndrome,the use of bulleyaconitine A tablets combined with Jiaoyan Qizi hot package can alleviate knee joint pain,improve knee joint mobility,and enhance patients'quality of life.

Objective: To explore the expression of collagen type 1 alpha 2 (COL1A2) in cervical cancer and its correlation with immune infiltration. Methods: Bioinformatics techniques were used to analyze the expression of COL1A2 in cervical cancer. Western blot and RT-qPCR were used to detect the expression of COL1A2 in cervical cancer tissues and cell lines. The correlation between the expression of COL1A2 and tumor immune cell infiltration was analyzed by tumor immune estimation resource (TIMER2. 0) . Gene set enrichment analysis (GSEA) was used to analyze the possible mechanism of COL1A2 in cervical cancer. Jaspar database was used to predict the transcrip- tion factors of COL1A2. Western blot and RT-qPCR were used to detect the expression of transcription factors in cervical cancer tissues and cell lines. Results: The expression of COL1A2 was down-regulated in cervical cancer (P < 0. 05) . The expression of COL1A2 was positively correlated with the levels of macrophages and myeloid den- dritic cells (P < 0. 01) . The proportions of 22 types of immune cells were different in different cervical cancer pa- tients. In addition , compared with the high expression group of COL1A2 , the proportion of M0 macrophages , M2 macrophages and resting memory CD4 + T cells increased in the low expression group of COL1A2 , while the propor- tion of CD8 + T cells , activated memory CD4 + T cells , follicular helper T cells , activated NK cells and activated myeloid dendritic cells decreased (P < 0. 05) . GSEA analysis showed that COL1A2 was related to immune-related signaling pathways , including Notch signaling pathway , interleukin-6/janus kinase/signal transducer and activator of transcription 3 (IL6/JAK/STAT3) , Wnt/β-catenin signaling pathway , etc. (P < 0. 01) . Jaspar database pre- dicted that the transcription factor of COL1A2 was paired box protein 5 (PAX5) , and the expression of PAX5 de- creased in cervical cancer (P < 0. 05) . Conclusion COL1A2 is expected to become a potential diagnostic biomar- ker and immunotherapy target for cervical cancer.

Objective:To analyze the epidemiological, etiological and genetic characteristics of influenza virus in Shandong Province during 2023-2024.Methods:The surveillance data of influenza-like illness (ILI) in sentinel hospitals in Shandong from 2023 to 2024 were collected and analyzed. The isolated influenza strains with hemagglutination titers ≥8 were selected for antigenicity analysis, drug susceptibility test, gene sequencing and evolutionary analysis.Results:From 2023 to 2024, the positive rate of influenza virus in Shandong was 8.51% (23 663/277 995), the highest positive rate was in the age group of 5-14 years (15.78%, 6 073/38 478), and the highest positive rate was in the 49 th week (35.86%, 2 264/6 313). Both antigenicity analysis and evolutionary analysis showed that the A(H1N1)pdm09 subtype and B(Victoria) strain had good matching effect and close evolutionary distance with the 2023-2024 surveillance year vaccine strain. The A(H3N2) subtype strain did not have a high matching effect with the 2023-2024 vaccine strain and had a long evolutionary distance, but had a close evolutionary distance with the 2024-2025 vaccine strain. Drug susceptibility test showed that oseltamivir sensitivity of influenza A(H1N1)pdm09 strain decreased greatly, and the amino acid site mutation of neuraminidase was H275Y. Conclusions:In the 2023-2024 surveillance year, the peak of influenza virus epidemic in Shandong was mainly occurred in winter and spring, and the age group of 5-14 years was the focus of prevention and control. The dominant strain was subtype A(H3N2), which had poor matching effect with the vaccine strain in the 2023-2024 surveillance year. One A(H1N1)pdm09 resistant strain was found in the drug resistance monitoring work. Follow-up prevention and control work should be strengthen the surveillance for the epidemiological characteristics, genetic variation and drug resistance of influenza viruses, timely understand the epidemic trend and mutation of influenza viruses, timely discover drug-resistant strains of influenza viruses, promote influenza vaccination, and improve of influenza prevention and control.

Cyclic neutropenia (CN) is relatively rare in clinical practice, and cases complicate with intestinal fistula have not been reported yet. This article reports a 45 year old female patient with CN and intestinal fistula, hoping to improve clinical experience in the diagnosis and treatment of such diseases.

Objective:To analyze the respiratory virus infection status and epidemiological characteristics of influenza-like illness (ILI) cases in Shandong Province during the 2020 -2021 influenza surveillance year. Methods:According to the National Influenza Surveillance Plan (2017 version), throat swab samples of ILI cases were collected from 14 surveillance sentinel hospitals in Shandong Province. Nucleic acid was extracted from all samples. Real-time fluorescence quantitative PCR (RT-PCR) was utilized to detect six common viruses, including human metapneumovirus (HMPV), human parainfluenza virus (HPIV) types 1, 2 and 3, respiratory syncytial virus (RSV), and adenovirus (ADV). Subsequently, the obtained detection results were analyzed.Results:A total of 2 386 specimens were collected, with a detection rate of 24.22% (578). Six viruses were detected, with detection rates of 6.75% (162 cases) for HMPV, 5.87% (140 cases) for RSV, 3.56% (85 cases) for HPIV3, 3.14% (75 cases) for HPIV2, 2.98% (71 cases) for HPIV1, and 2.77% (66 cases) for ADV. There was no significant difference in detection rates between genders, but a notable variation among different age groups ( P<0.001). The highest detection rate was observed in individuals aged 0-4 years (31.94%), followed by those aged≥60 years (26.06%). The prevalence of six viruses showed a monthly variation, with the detection rate of HMPV being higher in December and HPIV1 being higher in February. HPIV2, HPIV3, RSV, and ADV had higher detection rates in November. The co-detection rate of multiple viruses was 0.80%, with RSV being the most common pathogen involved in co-detection, primarily in individuals aged 0-4 years. Conclusion:The detection of six multiple pathogens in ILI cases in Shandong Province is dominated by HMPV, RSV and HPIV3. The prevalence of respiratory viruses varies by age and time.

This study investigated the epidemic status of H3N2 influenza virus and the genetic evolution characteristics of hemagglutinin(HA)and neuraminidase(NA)of H3N2 subtype influenza viruses isolated in Shandong Province during 2023-2024,to understand their compatibility with vaccine strains and drug resistance status.A total of 25 H3N2 subtype influenza virus strains were randomly selected from the strains isolated by the influenza surveillance network laboratory.The HA and NA genes were sequenced with the vaccine strains recommended by the WHO as a reference.Monitoring of sensitivity to oseltamivir and zanamivir was conducted through neuraminidase inhibition experiments.The H3N2 influenza viruses in Shandong Province belonged to the 3C.2a1b.2a.2a.3a.1 clade.Nucleotide sequence analysis revealed that the HA1 and NA genes were closely related to the WHO-recommended vaccine strain A/Darwin/9/2021 for the current season,with homology rates of 97.8%-98.2%and 98.9%-99.3%,respectively.Amino acid sequence analysis indicated 22 amino acid sequence variations in the HA1 protein,and antigenic drift was detected in 8 strains.A glycosylation site was added at position 94 of the HA protein in all 25 strains.Variations occurred in the NA antigenic determinants of some strains.Neuraminidase inhibition experiments indicated that all tested influenza viruses were sensitive to oseltamivir and zanamivir.Some differences in HA and NA proteins were observed between the monitored strains and vaccine strains.Continued monitoring of the molecular evolution characteristics of influenza viruses is necessary to understand the risk of influenza outbreaks,and their effects on the effectiveness of influenza vaccines and therapeutic drugs.

Objective:To investigate the extent and characteristics of parenting stress experienced by fathers of children aged 0-6 in three provinces of Eastern China, and to compare the quality of their marital relationships within each latent class, in order to provide reference for developing intervention strategies for different categories of fathers.Methods:Using cross-sectional survey methods, fathers with children aged 0-6 years were recruited using convenience sampling from December 2020 to March 2021 from Anhui province, Zhejiang province and Jiangsu province in Eastern China. Data were collected through a general information questionnaire, the Parenting Stress Index-Short Form, and the Locke-Wallace Marital Adjustment Scale.Latent profile analysis was used to determine the classification, Kruskal-Wallis H test was used to compare the difference of marital quality. Results:A total of 657 questionnaires were distributed and 604 valid questionnaires were recovered. The age of 604 fathers was (32.50 ± 6.26) years, the score of parenting stress was (89.70 ± 17.87) and the score of marriage quality was (108.36 ± 22.28). The characteristics of parenting stress among fathers of young children were categorized into three latent profile types: "high stress-negative parenting group" (88 people accounting for 14.6%), " medium stress- poor interaction group" (329 people accounting for 54.5%), and " low stress-positive parenting group" (187 people accounting for 30.9%). The scores of marriage quality were (99.94 ± 23.81), (106.52 ± 19.84), (115.56 ± 23.70) points among these three latent classes with significant differences ( H=35.89, P<0.05). Conclusions:The study identifies three latent classes of parenting stress among fathers of children aged 0-6, each associated with varying levels of marriage quality. Therefore, targeted interventions aimed at improving fathers′ parenting stress and marriage quality, and helping create family environments where children can grow up healthy.

Objective To construct mice hepatocellular carcinoma models with different tumor immune microenvironments(TIME)and explore the differences.Methods H22 and hepa1-6 were used to construct subcutaneous transplantation tumor model of C57 mice as homologous hepatocellular carcinoma cell lines(denoted as H22 group and hepal-6 group,each n=8),and the differences of TIME were evaluated.Immunohistochemistry was used to detect and quantify the infiltration of T cells,CD4+T cells,CD8+T cells,regulatory T cells and B cells in TIME.Flow cytometry was performed to detect the differences of composition of immune cell subpopulations in peripheral blood and tumor parenchyma.Gene expression profile characteristics of tumor tissue were analyzed based on high-throughput transcriptome sequencing technology,and enrichment analyses of immune-related signaling pathways were evaluated combined with gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG).Results H22 group showed cold and hepa1-6 group showed hot TIME characteristics.The number of T cells,CD4+T cells and CD8+T cells in tumor tissue of H22 group were all lower,while the proportion of T cells,CD4+T cells and CD8+T cells in peripheral blood were all higher than those of hepa1-6 group(all P＜0.05).Compared with H22 group,up-regulated genes of tumor tissue in hepa1-6 group expressed significantly enriched in tumor immune activation-related signaling pathways.Conclusion H22 and hepa1-6 hepatocellular carcinoma models showed distinct TIME characteristics of cold and hot tumors,respectively,and the amount of immune cells in tumor tissue of the former were significantly lower than those in the latter.

Objective:To analyze the epidemiological, etiological and genetic characteristics of influenza virus in Shandong Province during 2023-2024.Methods:The surveillance data of influenza-like illness (ILI) in sentinel hospitals in Shandong from 2023 to 2024 were collected and analyzed. The isolated influenza strains with hemagglutination titers ≥8 were selected for antigenicity analysis, drug susceptibility test, gene sequencing and evolutionary analysis.Results:From 2023 to 2024, the positive rate of influenza virus in Shandong was 8.51% (23 663/277 995), the highest positive rate was in the age group of 5-14 years (15.78%, 6 073/38 478), and the highest positive rate was in the 49 th week (35.86%, 2 264/6 313). Both antigenicity analysis and evolutionary analysis showed that the A(H1N1)pdm09 subtype and B(Victoria) strain had good matching effect and close evolutionary distance with the 2023-2024 surveillance year vaccine strain. The A(H3N2) subtype strain did not have a high matching effect with the 2023-2024 vaccine strain and had a long evolutionary distance, but had a close evolutionary distance with the 2024-2025 vaccine strain. Drug susceptibility test showed that oseltamivir sensitivity of influenza A(H1N1)pdm09 strain decreased greatly, and the amino acid site mutation of neuraminidase was H275Y. Conclusions:In the 2023-2024 surveillance year, the peak of influenza virus epidemic in Shandong was mainly occurred in winter and spring, and the age group of 5-14 years was the focus of prevention and control. The dominant strain was subtype A(H3N2), which had poor matching effect with the vaccine strain in the 2023-2024 surveillance year. One A(H1N1)pdm09 resistant strain was found in the drug resistance monitoring work. Follow-up prevention and control work should be strengthen the surveillance for the epidemiological characteristics, genetic variation and drug resistance of influenza viruses, timely understand the epidemic trend and mutation of influenza viruses, timely discover drug-resistant strains of influenza viruses, promote influenza vaccination, and improve of influenza prevention and control.

This study investigated the epidemic status of H3N2 influenza virus and the genetic evolution characteristics of hemagglutinin(HA)and neuraminidase(NA)of H3N2 subtype influenza viruses isolated in Shandong Province during 2023-2024,to understand their compatibility with vaccine strains and drug resistance status.A total of 25 H3N2 subtype influenza virus strains were randomly selected from the strains isolated by the influenza surveillance network laboratory.The HA and NA genes were sequenced with the vaccine strains recommended by the WHO as a reference.Monitoring of sensitivity to oseltamivir and zanamivir was conducted through neuraminidase inhibition experiments.The H3N2 influenza viruses in Shandong Province belonged to the 3C.2a1b.2a.2a.3a.1 clade.Nucleotide sequence analysis revealed that the HA1 and NA genes were closely related to the WHO-recommended vaccine strain A/Darwin/9/2021 for the current season,with homology rates of 97.8%-98.2%and 98.9%-99.3%,respectively.Amino acid sequence analysis indicated 22 amino acid sequence variations in the HA1 protein,and antigenic drift was detected in 8 strains.A glycosylation site was added at position 94 of the HA protein in all 25 strains.Variations occurred in the NA antigenic determinants of some strains.Neuraminidase inhibition experiments indicated that all tested influenza viruses were sensitive to oseltamivir and zanamivir.Some differences in HA and NA proteins were observed between the monitored strains and vaccine strains.Continued monitoring of the molecular evolution characteristics of influenza viruses is necessary to understand the risk of influenza outbreaks,and their effects on the effectiveness of influenza vaccines and therapeutic drugs.