ObjectiveTo investigate the therapeutic effect of Astragali Radix-mediated changes in gut microbiota on treating type 2 diabetes （T2DM）. MethodsA 12-week randomized， placebo-controlled clinical trial enrolled eighty patients with newly diagnosed type 2 diabetes and poor glycemic control in the Qi deficiency type. All patients received insulin therapy. The observation group （40 cases） was administered with Astragali Radix Granules， while the control group （40 cases） received a placebo. Both treamtents were taken orally twice daily. Changes in gut microbiota were assessed by 16s rDNA sequencing. Serum glucagon-like peptide-1 （GLP-1） levels were measured using enzyme-linked immunosorbent assay （ELISA）. Glucose metabolism indicators including fasting blood glucose （FPG）， 2-hour postprandial blood glucose （2 h PG），glycated albumin（GA）， and glycated hemoglobin （HbA1c） were evaluated. Pancreatic function was evaluated using fasting C-peptide （FCP）， 2-hour postprandial C-peptide （2 h CP）， and C-peptide area under the curve （AUC_cp）. Traditional Chinese medicine （TCM） syndrome scores， clinical efficacy， and safety indicators were also observed. ResultsIn terms of glucose metabolism indicators， compared with the baseline， both groups exhibited significantly lower FPG， 2 h PG， GA and HbA1C （P<0.01），while FCP， 2 h CP and AUC_cp were significantly higher （P<0.01）. Compared with the control group after the treatment， the observation group showed significantly lower FPG， 2 h PG， GA and HbA1C（P<0.05， P<0.01），and significantly higher FCP， 2 h CP and AUC_cp （P<0.05， P<0.01）， indicating that Astragali Radix can improve glucose metabolism. In terms of the diversity of gut microbiota， no significant differences were detected in the Chao1， Shannon and Simpson indexes of the two groups compared with their respective baselines. However， compared with the post-treatment control group， the observation group demonstrated significant increases in the Chao1， Shannon and Simpson indexes （P<0.05， P<0.01）. The β-diversity analysis showed significant separation in gut microbiota composition before and after treatment in both groups， indicating that Astragali Radix can significantly alter the structure and improve the diversity of gut microbiota. At the phylum level， compared with the baseline， both groups showed a significant increase in the relative abundance of Bacteroidota（P<0.01）. The relative abundance of the potentially harmful phylum Proteobacteria was significantly lower in the observation Group after treatment （P<0.01）. Compared with the post-treatment control group， the observation group had a significantly higher relative abundance of Bacteroidota（P<0.01）. No significant difference was found in Firmicutes/Bacteroidota （F/B） ratio between the two groups after treatment， and other phyla showed no significant differences. At the genus level， compared with the baseline， the observation group exhibited a significant increase in Bacteroides （P<0.01） and a significant decrease in Escherichia-Shigella （P<0.01）， whereas no significant difference was seen in the control group . Compared with the control group after treatment， the observation group after treatment had a significantly higher relative abundance of Bacteroides （P<0.01）. No significant differences were seen in other genera. Linear discriminant analysis （LDA） identified potential characteristics taxa： in the observation group， Bacteroidota at the phylum level and Bacteroides and Dubosiella at the genus level， in the control group， Proteobacteria at the phylum level as well as Barnesiella and Staphylococcus at the genus level. Correlation analysis based on a heatmap revealed that GLP-1 levels were positively correlated with Firmicutes， F/B ratio and Fusobacterium， and negatively correlated with Bacteroidota， Proteobacteria， Bacteroides and Escherichia-Shigella. In terms of clinical efficacy， compared with the control group， the total effective rate of the observation group was significantly higher （P<0.05）. Compared with the baseline， the scores for shortness of breath， fatigue， weakness， spontaneous sweating and reluctance to speak significantly decreased in both groups （P<0.01）. Compared with the control group after treatment， the score for weakness was significantly lower in the observation group （P<0.01），indicating that Astragali Radix could improve clinical symptoms and alleviate weakness symptoms. In terms of safety， compared with the baseline， alanine aminotransferase （ALT） levels significantly decreased in both groups （P<0.05，P<0.01），indicating that Astragali Radix did not induce any significant abnormalities in liver and kidney functions. ConclusionAstragali Radix demonstrates the potential to significantly improve the gut microbiota environment in patients of newly diagnosed type 2 diabetes with Qi deficiency. The therapeutic effect may contribute to glycemic control， possibly mediated by an elevation in GLP-1 level. These findings may support its further clinical investigations and potential applications.

ObjectiveTo investigate the effects of Liuwei Dihuangwan on mammalian target of rapamycin （mTOR）-mediated synaptic plasticity in the prefrontal cortex-hippocampus circuit in the rat model of valproic acid （VPA）-induced autism spectrum disorder （ASD） and to explore its potential mechanism for reducing behavioral abnormalities in ASD rats. MethodsThirteen SD pregnant rats were randomized into the VPA group （10 rats） and the control group （3 rats）. On gestational day 13， the VPA group was intraperitoneally injected with VPA solution （600 mg·kg^-1）， while the control group was injected with an equal volume of normal saline. Male pups of 21 days old from both groups underwent behavioral tests to screen the ASD model rats （32 rats）. These rats were then randomly assigned to the model （distilled water）， vitamin D （1 480 U·kg^-1）， low-dose （0.75 g·kg^-1） Liuwei Dihuangwan， and high-dose （1.5 g·kg^-1） Liuwei Dihuangwan groups， and normal male pups were selected as the blank group （distilled water）， with 8 rats in each group. Each group was treated with the corresponding drug or an equal volume of distilled water via gavage daily. After drug administration， behavioral tests were conducted again， followed by anesthesia and tissue collection. Nissl staining was carried out to observe the morphological changes of neurons in the prefrontal cortex and hippocampus. Transmission electron microscopy was used to observe changes in hippocampal synaptic structure. Immunohistochemistry and Western blot were employed to determine the expression of mTOR， phosphatidylinositol 3-kinase （PI3K）， and protein kinase B （Akt） in the prefrontal cortex， as well as synaptophysin-1 （SYP-1）， postsynaptic density 95 （PSD95）， and N-methyl-D-aspartate receptors （NMDARs） in the hippocampus. ResultsCompared with the control group， the model group exhibited severe growth retardation and developmental deformities， with abnormal behavioral performance and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus. Compared with the model group， all treatment groups showed reduced behavioral abnormalities and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus， with the most significant effects in the high-dose Liuwei Dihuangwan group. Immunohistochemistry and Western blotting results showed that compared with the control group， the model group exhibited significantly increased protein levels and phosphorylation ratios of p-mTOR， p-Akt， and p-PI3K in the prefrontal cortex （P<0.05， P<0.01）， and decreased protein levels of SYP-1 and PSD95 and increased protein level of NMDARs in the hippocampus （P<0.01）. Compared with those in the model group， the protein levels of p-mTOR， p-Akt， and p-PI3K in the prefrontal cortex decreased in the vitamin D and high-dose Liuwei Dihuangwan groups （P<0.05）， and the p-mTOR/mTOR， p-Akt/Akt， and p-PI3K/PI3K ratios in the prefrontal cortex declined in the vitamin D group （P<0.05）. The high-dose Liuwei Dihuangwan group showed a reduction in p-PI3K/PI3K ratio （P<0.05）. The protein level of SYP-1 and PSD95 in the hippocampus increased in the vitamin D， low-dose Liuwei Dihuangwan， and high-dose Liuwei Dihuangwan groups （P<0.05， P<0.01）， while the protein level of NMDARs decreased in the vitamin D and high-dose Liuwei Dihuangwan groups （P<0.01）. ConclusionLiuwei Dihuangwan significantly reduces VPA-induced ASD-like behaviors and enhances the regeneration and repair of neurons in the prefrontal cortex and synaptic structures in the hippocampus by downregulating of PI3K/Akt/mTOR expression in the prefrontal cortex， upregulating SYP-1 and PSD95 expression in the hippocampus， and inhibiting NMDARs expression.

A solid-phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry(SPE-UPLC-MS/MS)method was established for simultaneous determination of 10 kinds of neonicotinoid pesticide residues in aquaculture water.Based on the chemical properties of neonicotinoid pesticides and the matrix characteristics of aquaculture water,suitable temporary storage methods for water samples and appropriate solid-phase extraction columns were selected,and the extraction conditions(including elution solvents and sample loading volumes)were optimized.The method employed acetonitrile and 5 mmol/L ammonium acetate solution(containing 0.1%formic acid)as the mobile phase and an Oasis HLB solid-phase extraction column combined with PSA as a dispersive sorbent for sample purification.The method exhibited good linearity in detection of neonicotinoid in concentration range of 0.2-50 ng/mL(R2>0.99797),with a detection limit of 0.5 ng/L and a quantification limit of 1 ng/L,which were significantly lower than the maximum acceptable method detection limits(9-500 ng/L)for neonicotinoid insecticides in water published by the European Commission.In pond water,rice-fish water,and seawater,the average recoveries of the 10 target analytes were 74.6%-114.1%,with relative standard deviations ranging from 0.3%to 9.6%.Using this method,actual sample tests were conducted on the Pearl River water,Zhaoqing pond water,and Qingyuan rice-fish aquaculture water.The total concentration of five neonicotinoid pesticides in the Pearl River water ranged from 154.8 to 246.6 ng/L,the total concentration of four neonicotinoid pesticides in the Zhaoqing pond water was 95.0-176.1 ng/L,and the total concentration of three neonicotinoid pesticides in the Qingyuan rice-fish aquaculture water was 2.3-11.7 ng/L.This method was simple in operation,highly sensitive,and had strong resistance to interference.It was suitable for detection of trace neonicotinoid pesticides in aquaculture water and could provide technical support for construction of a green aquaculture environment and resolution of international trade disputes.

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.

OBJECTIVE To investigate the pharmacodynamics and related mechanisms of Anmei Dan on hippocampus of mice with coronary heart disease complicated with depression.METHODS The coronary heart disease model combined with chronic and unpredictable mild stress depression model was established,and the mice were randomly divided into blank group,model group,low dose group(1.5 g·kg-1),high dose group(3 g·kg-1)and atorvastatin group(0.3 g·kg-1).Sucrose preference test,open field test and forced swimming test were used to evaluate the behavioral changes of mice.qPCR and ELISA were employed to ascertain the mR-NA expression of interleukin-1β(IL-1β),interleukin-6(IL-6)and tumour necrosis factor-α(TNF-α)in the hippocampus.The changes of neurons and Nissl bodies in CA1,CA3 and DG regions of hippocampus were observed by Nissl staining.The expression of key proteins was detected by Western blot.RESULTS Compared with the blank group,the sucrose preference rate of mice in the model group was decreased(P<0.01),the forced swimming immobility time was extended(P<0.01)and the movement distance in the open field experiment was not significantly changed.The levels of total cholesterol(TC),triglyceride(TG)and low density lipopro-tein cholesterol(LDL-C)were increased significantly(P<0.01)and the level of high density lipoprotein cholesterol(HDL-C)was decreased significantly(P<0.01).The mRNA level and content of IL-1β,IL-6 and TNF-α were significantly increased(P<0.01).The expression of glutamate receptor 1(GluR1),postsynaptic densitin-95(PSD-95),brain-derived neurotrophic factor(BDNF)and phosphorylated calmodulin-dependent kinase(p-CaMKⅡ)in hippocampus was decreased(P<0.01).The expression of cytoskeletal activity regulatory protein(Arc)was increased(P<0.01).In the model group,the cell structure was irregular and differ-ent degrees of damage occurred,the Nissl bodies decreased or disappeared,and the cell membrane broke.Compared with the model group,the sucrose preference rate of mice in each administration group was significantly increased(P<0.01),the immobility time in forced swimming experiment was significantly decreased(P<0.01),and the levels of TC,TG and LDL-C of mice in Anmei Dan groups and atorvastatin group were decreased(P<0.01),while the level of HDL-C increased(P<0.01).The mRNA levels and con-tent of IL-1β,IL-6 and TNF-α in Anmei Dan groups and atorvastatin group were decreased(P<0.01).The expression of GluR1,PSD-95,BDNF and p-CaMKⅡ in hippocampal tissue of Anmei Dan groups were increased(P<0.01),and the expression of Arc was decreased(P<0.01).The morphology and structure of the cells in the Anmei Dan group and the atorvastatin group were improved,with varying degrees of increased Nissl bodies and relatively intact cell membranes.CONCLUSION Anmei Dan can effectively im-prove blood lipids and depression-like behavior of coronary heart disease mice complicated with depression.It can inhibit pro-inflam-matory factors,increase the expression of neurotrophic factors,effectively improve synaptic related proteins,and reduce the damage to neurons,thus effectively preventing the exacerbation of coronary heart disease and depression comorbidity.

Objective To investigate the potential categories of nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity,and analyze the relationship between various latent categories and dietary motivation,so as to provide references for clinical working staff to develop relevant dietary intervention strategies.Methods A convenient sampling method was used to enroll 270 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity from the endocrine department of a tertiary general hospital in Zhenjiang City,Jiangsu province,using basic information questionnaire,Nutrition Literacy Measurement Scale for Chinese adults,and the Regulation of Eating Behaviors Scale to collect data,using latent profile analysis to extract nutrition literacy potential categories,with general data and potential categories as independent variables,dietary motivation as the dependent variable for univariate analysis and multiple linear regression analysis.Results A total of 262 young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity completed the survey,and their nutrition literacy could be divided into 3 potential categories:low nutrition literacy group(16.41％),transformation dilemma group(54.96％),and middle and high nutrition literacy group(28.63％).The results of multiple linear regression analysis showed that residency style,potential categories of nutrition literacy and presence of diabetic complications were the influencing factors of dietary motivation in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity(P＜0.05).Conclusion There was a group heterogeneity in nutrition literacy in young and middle-aged patients with type 2 diabetes mellitus combined with overweight or obesity.Clinical working staff should provide targeted interventions based on the patient's potential categories of nutrition literacy to stimulate ongoing motivation to maintain a healthy diet.

Objective:To explore the application effect of visual eye dressings in patients after bilateral strabismus surgery.Methods:A total of 60 patients who underwent bilateral strabismus surgery in the Department of Ophthalmology of Beijing Tongren Hospital, Capital Medical University, from March to August 2024, were selected by convenience sampling. The patients were randomly divided into an observation group and a control group using a random number table method, with 30 cases in each group. After surgery, the control group was treated with conventional eye dressings, while the observation group was treated with visual eye dressings. The daily living ability, unplanned dressing changes were compared between the two groups.Results:The observation group had higher daily living ability scores and fewer unplanned dressing changes than the control group, and the differences were statistically significant ( P<0.05) . Conclusions:The use of visual eye dressings in patients after bilateral strabismus surgery has a positive effect on maintaining daily living ability and reducing the frequency of unplanned dressing changes.

Objective:To improve the diagnosis and treatment of autoimmune hepatitis (AIH) in children by analyzing their clinical characteristics and outcomes.Methods:Case summary.The general data, clinical manifestations, laboratory parameters, liver pathology, treatment and outcomes of 6 AIH children diagnosed at Xi′an Children′s Hospital from January 2019 to March 2024 were retrospectively analyzed.Results:All the 6 patients were diagnosed with AIH-1, and 5 of them were female, accounting for 83.3%.The median age of onset was 6.2 years old.Three cases (50.0%) had insidious onset.Elevated hepatic aminotransferase levels were detected incidentally in 2 cases (33.3%).One case (16.7%) presented symptoms resembling those of viral hepatitis.Two cases (33.3%) were accompanied by extrahepatic autoimmune diseases.All the children tested positive for anti-nuclear antibodies with a titer ≥1∶80 or 1∶100.Four cases (66.7%), 3 cases (50.0%), and 1 case (16.7%) tested positive for anti-smooth muscle antibodies, anti-perinuclear neutrophil cytoplasm antibodies and anti-soluble liver antigen, respectively.Liver pathology showed infiltration of lymphocytes-plasma cells and moderate or severe interfacial inflammation in all the 6 cases.Two cases (33.3%) had rosettes.All the patients were given oral corticosteroids combined with Azathioprine or Mycophenolate mofetil.The patients were followed up for 24.8 (12.0-52.0) months, during which 3 cases (50.0%) had partial remission and 3 cases (50.0%) achieved complete remission.Four cases (66.7%) showed transaminase fluctuations during corticosteroid reduction or cessation.Conclusions:Autoantibodies and liver pathology are important evidence for diagnosing AIH in children.Immunosuppressants have a high remission rate in the treatment of this disease, but transaminase fluctuations are prone to occur during corticosteroid reduction or cessation.

Objective To explore the utility of comprehensive geriatric assessment (CGA) in screening risk factors for osteosarcopenia (OS) among older adults (≥80 years old) and to evaluate the therapeutic efficacy of CGA-guided integrated interventions for OS. Methods A total of 420 patients aged ≥80 years, recruited from the Department of Geriatrics, General Practice of The Affiliated Jiangyin Hospital of Nantong University, and community health centers from January 2022 to October 2024, were enrolled. Participants were classified into OS (n＝139) and non-OS (n＝281) groups based on diagnostic criteria. CGA was utilized to compare differences in general characteristics, laboratory indicators, comorbidities between groups. Binary logistic regression analysis identified independent risk and protective factors. Subsequently, 40 OS patients were randomly assigned to an intervention group (n＝20) receiving integrated interventions including nutritional support, exercise training, and psychological management or a control group (n＝20, receiving routine care). Appendicular skeletal muscle mass index (ASMI), grip strength, gait speed, and bone mineral density (BMD) T-score were compared between groups after 3 months. Results The prevalence of OS in this cohort was 33.1%. Compared to the non-OS group, the OS group exhibited significant differences in age, body mass index (BMI), smoking history, comorbidity index, concomitant medication, cognitive impairment, visual and hearing impairment, sleep disorders, depression, marital status, social participation, activities of daily living, nutritional risk, total cholesterol, uric acid, and constipation (P＜0.05). Logistic regression analysis identified age and comorbidity index as significant risk factors for OS, while BMI, married status, total cholesterol, and activities of daily living (assisted and independent) served as protective factors. The intervention group demonstrated significant improvements in grip strength, gait speed, BMD T-score, and male ASMI compared to controls (P＜0.05). Conclusions CGA demonstrates clinical utility in systematically identifying risk factors for OS in the old population. Multimodal interventions guided by CGA effectively improve musculoskeletal function in elderly OS patients.

The prelimbic cortex (PL) plays a critical role in processing both the sensory and affective components of pain. However, the underlying molecular mechanisms remain poorly understood. In this study, we observed a reduction in hyperpolarization-activated cation current (I_h) in layer V pyramidal neurons of the contralateral PL in a mouse model of spared nerve injury (SNI). The expression of hyperpolarization-activated cyclic nucleotide-gated 2 (HCN2) channels was also decreased in the contralateral PL. Conversely, microinjection of fisetin, a partial agonist of HCN2, produced both analgesic and anxiolytic effects. Additionally, we found that cyclin-dependent kinase 5 (CDK5) was activated in the contralateral PL, where it formed a complex with HCN2 and phosphorylated its C-terminus. Knockdown of CDK5 restored HCN2 expression and alleviated both pain hypersensitivity and anxiety-like behaviors. Collectively, these results indicate that CDK5-mediated dysfunction of HCN2 in the PL underlies nerve injury-induced mechanical hypersensitivity and anxiety.
Animals ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels/metabolism* ; Hyperalgesia/metabolism* ; Cyclin-Dependent Kinase 5/metabolism* ; Neuralgia/metabolism* ; Male ; Anxiety/metabolism* ; Mice ; Potassium Channels/metabolism* ; Mice, Inbred C57BL ; Disease Models, Animal ; Pyramidal Cells/metabolism*