ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

ObjectiveTo study the effects of applying different microbial fertilizers on the growth and rhizosphere soil environment of Codonopsis pilosula and provide a theoretical basis for ecological cultivation of this medicinal plant. MethodsSeven groups were designed， including CK （no application of microbial fertilizer）， T1 （Trichoderma longibrachiatum fertilizer）， T2 （Bacillus subtilis fertilizer）， T3 （Trichoderma viride fertilizer）， T4 （compound microbial fertilizer）， T5 （C. pilosula stems and leaves fermented with compound microbial fertilizer）， and T6 （Scutellaria baicalensis stems and leaves fermented with T. viride fertilizer）. The physiological indicators， yield， and quality of C. pilosula and the physicochemical properties， enzyme activities， and microbial diversity in the rhizosphere soil of different fertilizer treatments were measured. ResultsGroup T1 showed slight decreases in soluble protein content （SPC） and superoxide dismutase （SOD）. Groups T2-T6 showed increases in physiological indicators such as proline （Pro）， soluble solids content （SSC）， SPC， catalase （CAT）， and peroxidase （POD） and a decrease in malondialdehyde （MDA） in C. pilosula leaves. All the fertilizer treatments increased the yield of C. pilosula and the total polysaccharide content in the roots. T1， T2， T3， T4， and T5 increased the total flavonoid content in the roots. Meanwhile， T4 increased the total saponin content in the roots. All the fertilizer treatments reduced the pH and increased the electric conductivity （EC）， soil organic matter （SOM）， and alkaline nitrogen （AN） in the soil. T2 and T5 increased the available phosphorus （AP）， and T3， T4， T5， and T6 increased the available potassium （AK） in the soil. All the fertilizer treatments increased the activities of urease， sucrase， and CAT in the soil. Except that T1 decreased the bacterial diversity in the soil， other fertilizer treatments significantly increased bacterial and fungal diversity in the soil. Different fertilizer treatments significantly affected the composition of bacterial and fungal communities in the soil. At the phylum level， the dominant bacterial phyla included Proteobacteria， Acidobacteriota， and Bacteroideta， and the dominant fungal phyla were Ascomycota， Mortierellomycota， and unclassified_fungi in the rhizosphere soil of C. pilosula after bacterial fertilizer treatment. At the genus level， unclassified Gemmatimonadaceae， Sphingomonas， and unclassified Vicinamibacteraceae were the dominant bacterial genera， while unidentified， unclassified Fungi， and unclassified Sordariomycetes were the dominant fungal genera in the rhizosphere soil. The results of redundancy analysis indicated that the main physicochemical factors affecting changes of microbial communities in the rhizosphere soil of C. pilosula were pH， EC， AK， AN， AP， and soil organic matter （SOM） in the soil. The correlation heatmap showed that Bryobacter had significantly positive correlations with EC， AK， and AN. There was a significantly negative correlation between Fusarium and SOM. In summary， applying an appropriate amount of microbial fertilizer can promote the growth and improve the rhizosphere soil environment of C. pilosula. ConclusionThe compound microbial fertilizer and the C. pilosula stems and leaves fermented with compound microbial fertilizer can improve the soil nutrients， growth， development， yield， and quality of C. pilosula， and thus they can be applied to the artificial cultivation of C. pilosula.

Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

Extracellular vesicles (EVs) are membrane-bound vesicles secreted by cells, including exosomes, microvesicles, and apoptotic bodies, which play critical roles in intercellular communication, material transport, and signal transduction. In recent years, increasing evidence has highlighted the essential function of EVs in early embryo development. By carrying bioactive molecules such as proteins, nucleic acids (e.g., mRNA and miRNA), and lipids, EVs regulate embryonic gene expression, cell proliferation, differentiation, and the microenvironment. Studies have shown that EVs derived from various segments of the female reproductive tract can enhance embryonic developmental potential, improve embryo quality, and facilitate implantation. Additionally, EVs secreted by embryos themselves participate in intercellular communication and play pivotal roles during embryogenesis. This review summarizes recent advances in understanding the functions of EVs in early embryo development, discusses their roles in mediating cell-cell communication and regulating gene expression, and explores their potential applica-tions in reproductive medicine and clinical practice, offering new perspectives for optimizing assisted reproductive technologies.

Autism Spectrum Disorders (ASDs) are reported as a group of neurodevelopmental disorders. The structural changes of brain regions including the hippocampus were widely reported in autistic patients and mouse models with dysfunction of ASD risk genes, but the underlying mechanisms are not fully understood. Here, we report that deletion of Trio, a high-susceptibility gene of ASDs, causes a postnatal dentate gyrus (DG) hypoplasia with a zigzagged suprapyramidal blade, and the Trio-deficient mice display autism-like behaviors. The impaired morphogenesis of DG is mainly caused by disturbing the postnatal distribution of postmitotic granule cells (GCs), which further results in a migration deficit of neural progenitors. Furthermore, we reveal that Trio plays different roles in various excitatory neural cells by spatial transcriptomic sequencing, especially the role of regulating the migration of postmitotic GCs. In summary, our findings provide evidence of cellular mechanisms that Trio is involved in postnatal DG morphogenesis.
Animals ; Dentate Gyrus/metabolism* ; Mice ; Morphogenesis/physiology* ; Neurons/pathology* ; Cell Movement ; Mice, Inbred C57BL ; Autism Spectrum Disorder/pathology* ; Mice, Knockout ; Neural Stem Cells ; Male ; Neurogenesis ; Autistic Disorder/genetics*

OBJECTIVE To investigate α1-adrenergic receptors(α1-AR)distribution in mouse tissues and its function on dexmetomidine(DMED)induced toxic effects.METHODS ① Real-time fluorescence quantita-tive PCR was used to detect the relative expression of α1A-AR,α1B-AR,α1D-AR,α2A-AR,α2B-AR and α2C-AR mRNAs in the heart,apical potion of heart,lungs,apical potion of lung,liver,kidneys,abdominal aorta,prefrontal cortex,hippocampus,striatum,brainstem,thalamus,olfactory bulb,and the rest of the brain tissues of the mouse.The relative expression of mRNA were analyzed.② C57BL/6J mice were pretreated with α2 adrenergic receptor antagonist atipamezole ATI(0.005,0.010,0.020,0.025,0.040,0.050 mg·kg-1,im),orα1-adrenoceptor antagonist prazosin(1 mg·kg-1,im)for 15 min,and then DMED(0.20 mg·kg-1,iv)was given to observe the rate of the loss of righting reflex and the immobilization time in mice.③ C57BL/6J mice were treated with DMED(16.38,20.48,25.60,32.00,40.00,and 50.00 mg·kg-1,iv)to observe the lethality of the mice in 24 h.The dose-effect relationship curves of the lethality rate and the half lethal-dose(LD50)were detected.ATI(1,2,4,and 8 mg·kg-1,im)or prazosin(1 mg·kg-1,im)were pretreated 15 min followed by the administration of DMED(25.60 mg·kg-1,iv).The lethality of the mice were recorded for 24 h.HE staining to observe the lung tissue damage in the mice.RESULTS ① The mRNA expression levels of three α1-AR subtype were higher than those of α2-AR subtype.α2A-AR and α2C-AR were highly expressed in the central nervous system.α2B-AR was highly expressed in the brainstem and peripheral tissues.The mRNA expres-sion levels of α1-AR subtypes were higher than those of α2-AR subtypes in heart,apical potion of heart or lung(P＜0.05).② ATI(0.005 to 0.05 mg·kg-1,im)dose dependently antagonized the loss of righting reflex and decreased the immobilization time induced by DMED(0.20 mg·kg-1,iv).In contrast,prazosin(1 mg·kg-1,im)had no effect on the loss of righting reflex induced by DMED(0.20 mg·kg-1,iv).③ The LD50 of DMED in mice was 26.734 mg·kg-1(iv)with a 95％Cl of 23.606-30.000 mg·kg-1.DMED(25.6 mg·kg-1)was selected for subsequent toxicity.ATI(1,2,4,and 8 mg·kg-1,im)did not antagonize the lethality induced by DMED(25.6 mg·kg-1,iv).The high dose of ATI resulted in elevated death rate and accelerated mortality induced by DMED(25.6 mg·kg-1,iv)in the mice.However,prazosin(1 mg·kg-1,im)reduced the lethality of DMED(25.6 mg·kg-1,iv)(P＜0.01).After administration of DMED(25.6 mg·kg-1),the mice lungs showed significant congestion.HE staining of lung tissues revealed obvious vascular hemorrhage,alveolar rupture,and erythrocyte spillage.Prazosin(1 mg·kg-1,im)effectively attenuated the tissue damage in the lungs,but ATI(1 mg·kg-1,im)aggravated the pulmonary hemorrhage.CONCLU-SIONS In cardiopulmonary tissues,the high expression levels of α1 adrenoceptor overactivation,might related with the lethality induced by DMED.

Objective To explore the regularity of prescriptions for epidemic disease in Taiping Huimin Heji Ju Fang based on the latent structure model and association rules analysis,and to provide references for modern epidemic treatment.Methods Prescriptions for epidemic diseases were extracted from Taiping Huimin Heji Ju Fang.A high-frequency herb matrix(frequency≥10)was constructed,and high-frequency herbs were analyzed using MicrosoftExcel 2016,Lantern 5.0,and IBM SPSS Modeler 18.0 for efficacy classification,property/flavor/channel tropism statistics,latent structure modeling,and association rule analysis.Results Among the 200 collected herbal prescriptions,46 high-frequency medicinal materials were identified,such as licorice,fresh ginger,dried ginger,poriae,and ginseng.The top 3 drugs efficacy were tonifying deficiency,relieving surface and warming inner.The medicinal properties were mainly warm,and the medicinal flavors were pungent,bitter,and sweet.The meridian tropisms mainly included the spleen meridian,lung meridian and stomach meridian.The analysis of latent structure model suggested that there were 8 types of common symptoms of epidemic disease,such as wind evil attack exterior,wind-cold-dampness,wind-heat,excess-heat in triple energizer,dampness inhibits qi stagnation,yang deficiency,blockage and spleen-qi deficiency.Theanalysis of association rules obtained licorice-ephedra and licorice-atractylodes,which with a core of licorice medicine,and the 16 association rules such as dried ginger-cinnamon,pericarpium citri reticulatae-mangnolia officinalis and poriae-ginseng-atractylodes macrocephala after eliminating ginger,jujube and licorice.Conclusion Most of the medicines used in the treatment of epidemic diseases in Taiping Huimin Heji Ju Fang are pungent,bitter and dispelling evil,the treatment should follow the principle of dispelling evil,warming yang to dissipate cold,dispelling dampness,clearing heat and expelling fire,inducing resuscitation,supporting the right and supplementing deficiency,which embodies the treatment principle of dispelling evil and supporting right,and provides reference and ideas for the treatment of clinical diseases based on syndrome differentiation.

Objective:To investigate the characteristics of brain network reorganization in migraine patients and its relationship with the grading of right-to-left shunt (RLS) in patent foramen ovale(PFO) by integrated 3.0T structural magnetic resonance imaging (MRI),resting-state functional MRI(fMRI),and contrast-enhanced transthoracic echocardiography, thereby toexplore the mechanism underlying cognitive impairment in migraine.Methods:A total of 49 migraine patients (migraine group)and 16 demographically matched healthy subjects (healthy control group) who received diagnosis and treatment at China-Japan Union Hospital of Jilin University from September 2020 to September 2023 were selected as the study participants. Structural images (T1-weighted) and resting-state functional images were obtained from brain imaging (MRI and fMRI) data. The amplitude of low-frequency fluctuation (ALFF) algorithm was used to quantify the intensity of spontaneous neural activity in brain regions, while the Fazekas scale and the Age-Related White Matter Changes (ARWMC) scale were employed to assess white matter lesions. Participants were evaluated using the Mini-Mental State Examination (MMSE). Based on the results of contrast-enhanced transthoracic echocardiography (cTTE),the migraine group was further subdivided into A1 group (18 cases, no PFO), A2 group (17 cases, PFO with small-to-moderate RLS) and A3 group (14 case, PFO with large RLS) group, the differences in brain functional activity and cognitive function were compared among these groups.Results:By Fazekas scale scores and ARWMC scale scores, the incidence of hyperintense foci in the deep white matter regions in the migraine group were higher than those in the healthy control group: 67.3%(33/49) vs. 6/16, 61.2%(30/49) vs. 5/16, there were statistical differences ( P<0.05). Compared with the healthy control group, the migraine group exhibited higher spontaneous neural activity intensity in the brainstem,bilateral posterior cingulate cortex,left medial frontal gyrus,and left middle frontal gyrus,while showing reduced brain activity in the right angular gyrus region. The short-term delayed recall scores and total MMSE scores in the migraine group were lower than thosein the healthy control group: (1.61 ± 1.06) scores vs. (2.44 ± 0.81) scores, (25.06 ± 2.31) scores vs. (27.94 ± 1.44) scores, there were statistical differences ( P<0.05). The short-term delayed recall scores mong the A1 group, A2 group and A3 group had statistical difference( P<0.05). Conclusions:Migraine patients exhibit specific brain functional network reorganization and cognitive dysfunction, which are closely related to the degree of RLS.

Hot melt extrusion(HME)technology employs thermodynamic and kinetic principles to mix pharmaceutical polymers with crystalline drugs at high temperatures and extrude them,embedding drug molecules within the polymer matrix to form solid dispersions.Due to its solvent-free nature,capability for one-step processing,and support for continuous operation,HME has garnered significant attention in the pharmaceutical industry in recent years.This article introduced the basic principles and development history of HME technology and its marketed drugs.It reviewed the research progress of HME technology in improving drug solubility,masking taste,controlled release,targeted release,oral dispersible films,implant formulations,semi-solid formulations,and 3D printed formulations.Additionally,the article summarized the advantages and limitations of HME technology and provided an outlook on its future development.

Objective To summarize the current status of outcome indicators in randomized controlled trials of TCM intervention in pediatric myocarditis,to explore the existing problems,and to provide a reference for the construction of a core set of indicators for the treatment of pediatric myocarditis with TCM.Methods Randomized controlled trial literature on the treatment of pediatric myocarditis with TCM was retrieved from CNKI,Wanfang Data,VIP,SinoMed,PubMed,Embase,Cochrane Library and Web of Science from the establishment of the databases to 16th,Nov.2024.The basic characteristics,diagnostic criteria,TCM evidence,interventions and outcome indicators of the studies were extracted.The risk of bias was assessed for the selected studies using the Cochrane Collaboration's risk of bias tool(RoB 2.0),and the outcome indicators were statistically analyzed using Excel 2019.Results Finally,totally of 250 articles were included in the literature.The outcome indicators were counted to obtain 187 outcome indicators with a cumulative total of 1 540 occurrences,which were categorized into six indicator domains:symptoms and signs,physicochemical testing,TCM symptoms/signs,clinical efficacy evaluation,safety evaluation and quality of life evaluation;among them,the physicochemical testing indicators(112 types,926 times)had the highest frequency of occurrences,followed by the clinical efficacy evaluation(13 types.340 times),and safety evaluation(24 types,193 times).Conclusion The overall quality of randomized controlled trials of TCM treatment of pediatric myocarditis is low,and there are problems with primary and secondary differentiation of outcome indicators and lack of TCM characteristics.There is an urgent need to improve the core set of endpoint indicators that highlight the characteristics of TCM in order to improve the quality of clinical research.