Objective:To investigate, for multi-sensitized allergic rhinitis (AR) patients allergic to dust mites combined with other allergens (pollen, mold, animal dander, etc.), whether the single dust mite subcutaneous immunotherapy (SCIT) can improve the specific symptoms caused by other allergens in the patients, and to analyze the relationship between the effectiveness of symptom improvement in these patients and the type, quantity and severity of the allergens.Methods:A multicenter retrospective study was conducted to collect mul-sensitized AR patients from allergy or respiratory departments of 5 hospitals who received house dust mite allergen preparation SCIT for 12 to 36 months and met other inclusion and exclusion criteria from February to July 2024. General clinical data were collected and the perennial or seasonal symptoms before and after treatment were evaluated with visual analogue scale (VAS) to assess whether there was an perennial or allergen-specific symptom improvement (VAS score decrease ≥30%), by which the patients were divided into effective group and ineffective. R software was used to analyze the differences between groups by using Fisher′s exact test and Mann-Whitney U test. Results:A total of 62 patients were enrolled, and the treatment were effective in 39 of them, with an effective rate of 62.9%. For allergen-specific symptoms, the median age of the effective group was higher than that of the ineffective group (12 years old vs. 8 years old, P=0.039), and the effective rate in dust mite specific immunoglobin E (sIgE) grade ≤5 group was higher than that in sIgE grade >5 group (81.6% vs. 45.5%, P=0.008), and the effective rate of mold sIgE grade ≤2 group was higher than that of sIgE grade >2 group (83.3% vs. 28.6%, P=0.045), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). For perennial symptoms, the effective rate in the mold grade ≤2 group was higher than that in the sIgE grade >2 group (91.3% vs. 28.6%, P=0.010), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). There was no significant correlation between the treatment effectiveness of perennial or allergen-specific symptoms and the number of combined allergens, the grade of skin test, and the difference between the grade of combined allergens and that of dust mites ( P>0.05). Conclusion:Among the patients with multi-sensitized AR allergic to dust mites included in this study, single dust mite SCIT is effective in some of them, and for allergen-specific symptoms, the effective group was elder, and dust mite sIgE grade 6 and mold sIgE grade ≥2 was related to the low effective rate of SCIT. The present results are insufficient for selecting single or multiple AIT in any type of multi-sensitized patients.

Microalgae possess high photosynthetic efficiency, robust adaptability, and substantial biomass, serving as excellent biological resources for large-scale cultivation. Malic enzyme (ME), a ubiquitous metabolic enzyme in living organisms, catalyzes the decarboxylation of malate to produce pyruvate, CO₂, and NAD(P)H, playing a role in multiple metabolic pathways including energy metabolism, photosynthesis, respiration, and biosynthesis. In this study, we identified the Scenedesmus quadricauda malic enzyme gene (SqME) and its biological functions, aiming to provide excellent target genes for the genetic improvement of higher plants. Based on the RNA-seq data from S. quadricauda under the biofilm cultivation mode with high CO₂ and light energy transfer efficiency and small water use, a highly expressed gene (SqME) functionally annotated as ME was cloned. The physicochemical properties of the SqME-encoded protein were systematically analyzed by bioinformatics tools. The subcellular localization of SqME was determined via transient transformation in Nicotiana benthamiana leaves. The biological functions of SqME were identified via genetic transformation in Nicotiana tabacum, and the potential of SqME in the genetic improvement of higher plants was evaluated. The ORF of SqME was 1 770 bp, encoding 590 amino acid residues, and the encoded protein was located in chloroplasts. SqME was a NADP-ME, with the typical structural characteristics of ME. The ME activity in the transgenic N. tabacum plant was 1.8 folds of that in the wild-type control. Heterologous expression of SqME increased the content of chlorophyll a, chlorophyll b, and total chlorophyll by 20.9%, 26.9%, and 25.2%, respectively, compared with the control. The transgenic tobacco leaves showed an increase of 54.0% in the fluorescence parameter NPQ and a decrease of 30.1% in Fo compared with the control. Moreover, the biomass, total lipids, and soluble sugars in the transgenic tobacco leaves enhanced by 20.5%, 25.7%, and 9.5%, respectively. On the contrary, the starch and protein content in the transgenic tobacco leaves decreased by 22.4% and 12.2%, respectively. Collectively, the SqME-encoded protein exhibited a strong enzymatic activity. Heterologous expressing of SqME could significantly enhance photosynthetic protection, photosynthesis, and biomass accumulation in the host. Additionally, SqME can facilitate carbon metabolism remodeling in the host, driving more carbon flux towards lipid synthesis. Therefore, SqME can be applied in the genetic improvement of higher plants for enhancing photosynthetic carbon fixation and lipid accumulation. These findings provide scientific references for mining of functional genes from S. quadricauda and application of these genes in the genetic engineering of higher plants.
Nicotiana/genetics* ; Photosynthesis/physiology* ; Malate Dehydrogenase/biosynthesis* ; Plant Leaves/genetics* ; Scenedesmus/enzymology* ; Carbon Cycle/genetics* ; Lipid Metabolism/genetics* ; Plants, Genetically Modified/metabolism*

Objective:To investigate whether the levels of 15 micronutrients could influence the risk of HER2-positive and negative breast cancer using Mendelian randomization (MR) .Methods:In this study, multivariate Mendelian randomization analysis was used to obtain the data of 15 micronutrients (copper, calcium, carotene, folate, iron, magnesium, potassium, selenium, zinc, vitamin A, vitamin B12, vitamin B6, vitamin C, vitamin D and vitamin E) and HER2 (+/-) breast cancer in the Genome-Wide Association Study (GWAS) database. Analyses were performed using R software and the TwoSampleMR package. This study used the inverse variance weighting (IVW) method as the main method to analyze the causal relationship between micronutrients and HER2 (+/-) breast cancer, combined with MR-Egger regression, weighted median (WM) , Simple mode, and Weighted mode as complementary methods. Heterogeneity and horizontal pleiotropy of our results were detected by the Cochran Q test, MR-Egger regression test, MR-PRESSO composite test, and MR Egger intercept to enhance the reliability and stability of the results. Reverse MR analysis was performed as HER2 (+/-) breast cancer exposure factors. The micronutrients screened by the MR above analysis were used as outcome variables for effect and sensitivity analyses. Multivariate Mendelian randomization analysis was performed on the relevant trace elements screened by MR to avoid interference between variables. The independent effects of trace elements were analyzed to improve the reliability of our results.Results:The results of MR analysis showed that micronutrient vitamin B6 was negatively associated with the risk of developing HER2-negative breast cancer (IVM: OR=0.69,95% CI:0.45-0.96, P=0.029) . The results of the Cochran Q test, MR-Egger regression test, MR-PRESSO composite test, and MR Egger intercept test showed that there was no heterogeneity and horizontal pleiotropy among the instrumental variables. At the same time, the analysis of leave-one-out test confirmed that a single SNP had no significant effect on the overall results, which further enhanced the reliability and stability of the results. Conclusions:Among the 15 micronutrients, our results suggested that vitamin B6 may be a protective factor for HER-negative breast cancer. Our study provided a reference for the pathogenesis of HER-negative breast cancer and its early prevention and treatment.

Objective:To investigate whether the levels of 15 micronutrients could influence the risk of HER2-positive and negative breast cancer using Mendelian randomization (MR) .Methods:In this study, multivariate Mendelian randomization analysis was used to obtain the data of 15 micronutrients (copper, calcium, carotene, folate, iron, magnesium, potassium, selenium, zinc, vitamin A, vitamin B12, vitamin B6, vitamin C, vitamin D and vitamin E) and HER2 (+/-) breast cancer in the Genome-Wide Association Study (GWAS) database. Analyses were performed using R software and the TwoSampleMR package. This study used the inverse variance weighting (IVW) method as the main method to analyze the causal relationship between micronutrients and HER2 (+/-) breast cancer, combined with MR-Egger regression, weighted median (WM) , Simple mode, and Weighted mode as complementary methods. Heterogeneity and horizontal pleiotropy of our results were detected by the Cochran Q test, MR-Egger regression test, MR-PRESSO composite test, and MR Egger intercept to enhance the reliability and stability of the results. Reverse MR analysis was performed as HER2 (+/-) breast cancer exposure factors. The micronutrients screened by the MR above analysis were used as outcome variables for effect and sensitivity analyses. Multivariate Mendelian randomization analysis was performed on the relevant trace elements screened by MR to avoid interference between variables. The independent effects of trace elements were analyzed to improve the reliability of our results.Results:The results of MR analysis showed that micronutrient vitamin B6 was negatively associated with the risk of developing HER2-negative breast cancer (IVM: OR=0.69,95% CI:0.45-0.96, P=0.029) . The results of the Cochran Q test, MR-Egger regression test, MR-PRESSO composite test, and MR Egger intercept test showed that there was no heterogeneity and horizontal pleiotropy among the instrumental variables. At the same time, the analysis of leave-one-out test confirmed that a single SNP had no significant effect on the overall results, which further enhanced the reliability and stability of the results. Conclusions:Among the 15 micronutrients, our results suggested that vitamin B6 may be a protective factor for HER-negative breast cancer. Our study provided a reference for the pathogenesis of HER-negative breast cancer and its early prevention and treatment.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

This study aims to establish BHK-21 stable cell lines expressing APN from four species(human,pig,dog,and cat),the APN fragments were amplified from pEGFP-C1-APN plasmids of the four species stored in the laboratory to generate the recombinant plasmids pcDNA4.0-APN.Af-ter the recombinant plasmids were transfected into BHK-21 cells,the stable BHK-21 cell lines ex-pressing the APNs were selected by two rounds of limited dilution.The constructed BHK-21 cell lines were identified by indirect immunofluorescence assay(IFA),and their susceptibility to PD-CoV and TGEV was tested for these four cell lines.Virus infection experiments revealed that PD-CoV infected cells expressing human,pig,and dog APNs,while it did not infect cells expressing cat APN.Simultaneously,TGEV infected cells expressing pig,dog,and cat APNs,but did not infect cells expressing human APN.The results suggest that the risk of cross-species infection for different coronaviruses and the established cell line can be used effectively to evaluate the virus in-fection.The findings also revealed that PDCoV has the potential risk of cross-species infection of human and dog,and TGEV has the potential risk of cross-species infection of dog and cat.These results provide a basis for the prevention and control strategy of coronaviruses.

Objective:To investigate the effect of Isoorientin(Iso)on development of oral squamous cell carcinoma(OSCC)and the regulation on immune response mediated by cGAS-STING signaling pathway.Methods:CAL 27 cells were divided into control group,low,medium and high concentration Iso groups and high concentration Iso+cGAS inhibitor group(high concentration Iso+RU.521 group).Proliferation activity of CAL 27 cells was detected by MTT;TUNEL method was applied to detect apoptosis of CAL 27 cells;Transwell chamber test was applied to detect migration and invasion of CAL 27 cells;Western blot was applied to detect expressions of cGAS-STING pathway related proteins in CAL 27 cells;the tumor transplantation model of mice was prepared,and the tumor inhibi-tion rate was calculated.At the same time,peripheral blood was taken to detect the differentiation of T lymphocyte subsets in peripheral blood of mice in each group by flow cytometry.Results:Compared with control group,proliferation activity,numbers of migration and invasive cells in Iso treated groups were decreased,apoptosis rate,expressions of cGAS,STING protein,and phosphorylation expres-sions of TBK1,IRF3,tumor inhibition rate of mice,and percentages of CD4+T,CD8+T cells in peripheral blood were increased(P<0.05);further use of cGAS inhibitors reversed the inhibitory effect of Iso on development of OSCC and the promotion on cGAS-STING signaling pathway(P<0.05).Conclusion:Iso can inhibit the development of OSCC by activating the immune response mediated by cGAS-STING signaling pathway.

Objective:To investigate, for multi-sensitized allergic rhinitis (AR) patients allergic to dust mites combined with other allergens (pollen, mold, animal dander, etc.), whether the single dust mite subcutaneous immunotherapy (SCIT) can improve the specific symptoms caused by other allergens in the patients, and to analyze the relationship between the effectiveness of symptom improvement in these patients and the type, quantity and severity of the allergens.Methods:A multicenter retrospective study was conducted to collect mul-sensitized AR patients from allergy or respiratory departments of 5 hospitals who received house dust mite allergen preparation SCIT for 12 to 36 months and met other inclusion and exclusion criteria from February to July 2024. General clinical data were collected and the perennial or seasonal symptoms before and after treatment were evaluated with visual analogue scale (VAS) to assess whether there was an perennial or allergen-specific symptom improvement (VAS score decrease ≥30%), by which the patients were divided into effective group and ineffective. R software was used to analyze the differences between groups by using Fisher′s exact test and Mann-Whitney U test. Results:A total of 62 patients were enrolled, and the treatment were effective in 39 of them, with an effective rate of 62.9%. For allergen-specific symptoms, the median age of the effective group was higher than that of the ineffective group (12 years old vs. 8 years old, P=0.039), and the effective rate in dust mite specific immunoglobin E (sIgE) grade ≤5 group was higher than that in sIgE grade >5 group (81.6% vs. 45.5%, P=0.008), and the effective rate of mold sIgE grade ≤2 group was higher than that of sIgE grade >2 group (83.3% vs. 28.6%, P=0.045), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). For perennial symptoms, the effective rate in the mold grade ≤2 group was higher than that in the sIgE grade >2 group (91.3% vs. 28.6%, P=0.010), and there was no statistically significant correlation between the other allergen grades and the effective rate ( P>0.05). There was no significant correlation between the treatment effectiveness of perennial or allergen-specific symptoms and the number of combined allergens, the grade of skin test, and the difference between the grade of combined allergens and that of dust mites ( P>0.05). Conclusion:Among the patients with multi-sensitized AR allergic to dust mites included in this study, single dust mite SCIT is effective in some of them, and for allergen-specific symptoms, the effective group was elder, and dust mite sIgE grade 6 and mold sIgE grade ≥2 was related to the low effective rate of SCIT. The present results are insufficient for selecting single or multiple AIT in any type of multi-sensitized patients.

With the rapid development of generative artificial intelligence(GAI),its application in TCM research has attracted increasing attention.This article explored the application prospects of GAI in TCM research,analyzed its specific application scenarios in TCM clinical research,intelligent device development,and herbal medicine research,and identified the main challenges currently faced,including insufficient logical rigor,lack of high-quality clinical data,and shortage of interdisciplinary talents.This article proposed solutions such as constructing TCM knowledge graphs,standardizing TCM data collection,enhancing objective measurement and evaluation,and cultivating artificial intelligence talents with a background in TCM,in order to o provide references for the objective and intelligent research and application of TCM based on GAI,and further improve the clinical efficacy and quality of TCM.