Sepsis remains a challenge in critical care worldwide，with intestinal microbiome dysbiosis implicated in its onset，progression，and organ dysfunction. Germ-free （GF） and human microbiota-colonized animals，by eliminating confounding from resident microbiota and combining induction methods such as CLP/LPS/live-bacterial challenge with fecal microbiota transplantation （FMT） and mono- or combined colonization，enable causal tests and dissection of microbiota-host interactions and drug mechanisms. This review summarizes the application of GF and related models in sepsis research. At the immune level，TLR-related signaling and pathogen community-induced suppression of IRF3-dependent defense affect innate immunity and pathogen clearance. At the barrier level，reprogramming of tight junction proteins and increased permeability drive gut-derived amplification. At the metabolic level，short-chain fatty acids and bile acids function as metabolic valves influencing GPR/HDAC signaling. At the epigenetic level，EZH2 and histone deacetylation emerge as druggable targets. We further synthesize anti-sepsis evidence for traditional Chinese medicine （TCM），mainly from conventional animals and clinical studies，discussing how formulae （e.g.，Xuebijing，Shenfu，and Huanglian Jiedu Tang） and active constituents （e.g.，berberine，luteolin，ginsenosides，and resveratrol） may act through microbiota-related and direct host pathways，with FMT/metabolite replenishment enabling further causal testing. Finally，we propose a closed-loop GF-humanized-clinical stratification paradigm for integrated evaluation of microecology，epigenetics，immunity，and organ outcomes，providing a translational path for precision prevention and treatment based on TCM and the microbiome.

Vacuum compression molding (VCM) is a novel technology supporting the research and development of pharmaceutical solid dispersions. It is widely applied due to its precision and convenience in sample preparation. This technology integrates the principles of heating, melting, cooling, and vacuum compression to transform solid powders into shaped solids directly. By selecting different molds, temperatures, and pressures, researchers can prepare samples with diverse characteristics. This paper presents an overview of the equipment composition and working principles of VCM technology, demonstrating its distinct advantages in the formulation screening process of amorphous solid dispersions through comparative analysis with hot melt extrusion using case studies, and introduces its applications in the development of drug delivery systems and rheological characterization analysis, with a perspective on the future development of its functions.

Objective:To investigate the expression of chondroitin sulfate proteoglycan 4 pseudogene 12(CSPG4P12)in the small cell lung cancer(SCLC)tissue,its relationship with immune infiltration,and its effect on cell biological functions,and to clarify its effect in the occurrence and development of SCLC.Methods:The E-GEOD-60052 cohort was obtained by searching the ArrayExpress database for SCLC.The R language Bioconductor package was used to complete data filtering standardization,and 63 samples of SCLC tumor tissues and 7 samples of normal tissues were obtained.The Mann-Whitney U test was used to analyze the difference in CSPG4P12 expression levels between two groups.Pearson correlation analysis was used to evaluate the associations between CSPG4P12 expression levels and 47 immune checkpoint genes.The ESTIMATE algorithm and CIBERSORT algorithm were used to evaluate the correlations between CSPG4P12 expression and tumor immune cell infiltration.A case-control study was used to analyze the clinical data.A total of 230 patients with SCLC were selected as case group,and 230 healthy subjects were selected as control group.The genotyping of CSPG4P12 rs2880765,rs6496932 and rs8040855 was performed using TaqMan-MGB fluorescent probe labeling method.Odds ratio(OR)and 95％confidence interval(CI)were calculated by unconditional Logistic regression model to analyze the association between polymorphic genetic variation of CSPG4P12 gene and the risk of SCLC.The SCLC DMS114 cells were transfected with pUC-57 plasmid(control group)and CSPG4P12 over-expression plasmid(OV-CSPG4P12 group),respectively.The efficiencies of CSPG4P12 over-expression in two groups were verified by real-time fluorescence quantitative PCR(RT-qPCR)method.Cell counting kit-8(CCK-8)method was used to detect the cell proliferation activities of cells in two groups.Transwell chamber assay was used to detect the numbers of migration and invasion cells in two groups,respectively.Hoechst 33342 fluorescence staining was used to observe the cell apoptosis in two groups.Results:The ArrayExpress database E-GEOD-60052 cohort analysis showed that the expression level of CSPG4P12 mRNA in SCLC tissue was decreased compared with normal tissue(P＜0.001).The expression of CSPG4P12 had positive correlations with the immune checkpoint genes including leukocyte associated immunoglobulin like receptor 1(LAIR1)(r=0.47,P＜0.001),tumor necrosis factor(TNF)receptor superfamily member 9(TNFRSF9)(r=0.38,P＜0.01),and TNF superfamily member 9(TNFSF9)(r=0.44,P＜0.001).The ESTIMATE algorithm results showed that the matrix score,immune score and ESTIMATE composite score of the patients in CSPG4P12 low expression group were lower than those in CSPG4P12 high expression group(P＜0.01).The CIBERSORT algorithm results showed that compared with CSPG4P12 high expression group,the infiltration of M0 macrophages in CSPG4P12 low expression group was increased(P＜0.05)and the infiltration of mast cells resting was decreased(P＜0.05).The CSPG4P12 expression level had positive correlations with infiltration of mast cells resting(r=0.35,P=0.03)and mononuclear cell infiltration(r=0.34,P=0.034).In case-control studies,compared with AA genotype,CSPG4P12 rs2880765 AT and TT genotype carriers had a higher risk of SCLC(OR=1.68,95％CI=1.15-2.45,P＜0.01).The stratified analysis showed that genetic variation of rs2880765 A＞T increased the risk of SCLC in the male,younger age group(≤60 years)and smoking subgroups(males:OR=1.86,95％CI=1.18-2.93,P＜0.01;≤60 years:OR=1.73,95％CI=1.11-2.68,P＜0.01;smoking:OR=2.76,95％CI=1.49-5.13,P=0.001).The cell biology experiment showed that compared with control group,the proliferation abilities of the cells in OV-CSPG4P12 group were significantly decreased at 48 and 72 h(P＜0.01),while the number of migration cells at 24 h was significantly decreased(P＜0.01),the number of apoptotic cells at 24 h was increased(P＜0.05)and the number of invasion cells at 48 h was significantly decreased(P＜0.01).Conclusion:CSPG4P12 is lowly expressed in SCLC tumor tissue,which is associated with immune infiltration.The genetic variation of CSPG4P12 rs2880765 A＞T can increase the risk of SCLC,and its over-expression can inhibit cell proliferation,migration and invasion,and promote apoptosis.

AIM:To observe the effects of Huazhuo granules on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/Akt)/glycogen synthase kinase-3β(GSK-3β)and PI3K/Akt/forkhead box protein O1(FOXO1)signaling pathways in the liver tissue of type 2 diabetes mellitus(T2DM)rats,and to explore its mechanism of action in regulating glycogen synthesis and gluconeogenesis.METHODS:Thirty 8-week-old SPF-grade male SD rats were chosen,with 5 ran-domly selected ones as the blank control group.The remaining 25 rats were fed to a high-sugar and high-fat diet for 4 weeks,followed by a single intraperitoneal injection of streptozotocin to establish a T2DM rat model.After successful mod-eling,these rats were randomly divided into 5 additional groups:the model group,a positive control group treated with met-formin,and low,medium,and high-dose Huazhuo granule groups,resulting in a total of 6 groups with 5 rats in each.Rats in the treatment groups were gavaged with the corresponding doses of medication,while those in the blank control and model groups were gavaged with an equal volume of saline once daily for 4 consecutive weeks.After 24-hour fasting with free access to water,the rats were euthanized to measure blood glucose,serum insulin levels,and serum triglyceride(TG),total cholesterol(T-CHO),high-density lipoprotein cholesterol(HDL-C),aspartate aminotransferase(AST)and alanine aminotransferase(ALT)levels.Liver pathological morphology was assessed by hematoxylin-eosin staining,and liver glycogen storage was detected by periodic acid-Schiff staining.Immunohistochemistry and Western blot analysis were employed to detect the protein levels of p-PI3K,p-Akt,p-FOXO1 and p-GSK-3β in the liver of rats in each group.The mRNA levels of phosphoenolpyruvate carboxykinase(PEPCK),glucose-6-phosphatase(G6Pase),GSK-3β,FOXO1,PI3K and Akt in liver tissues were detected by RT-qPCR.RESULTS:Compared with the model group,the rats in the high-dose Huazhuo Granule group and the metformin group exhibited decreased blood glucose and insulin levels(P<0.01),reduced AST,ALT,TG,and T-CHO concentrations(P<0.01),and increased HDL-C concentrations(P<0.01).The phosphorylation levels of PI3K,Akt,FOXO1,and GSK-3β proteins were up-regulated(P<0.01).In con-trast,the gene expressions of G6Pase,PERCK,FOXO1,and GSK-3β were all down-regulated(P<0.01).Conversely,the gene expressions of PI3K and Akt were upregulated(P<0.01).Pathological morphology of the liver tissue improved,accompanied by increased glycogen deposition in hepatocytes.CONCLUSION:Huazhuo granule manifests effects in lowering blood glucose and serum insulin levels,ameliorating blood lipids,and enhancing liver function in rats.These ef-fects are revealed to increase glycogen synthesis by activating the PI3K/Akt/GSK-3β signaling pathway and reduce gluco-neogenesis by activating the PI3K/Akt/FOXO1 signaling pathway.

The clinical antiprotozoal drug nitazoxanide has been demonstrated to improve the experimental diabetes mellitus, lipid metabolism disorders, atherosclerosis and inhibit inflammation. Since the pathogenesis of heart failure with preserved ejection (HFpEF) is multifactorial and closely associated with the aforementioned diseases, we aim to study the effect of nitazoxanide on high-fat diet (HFD) plus L-NAME (N ω-nitro-l-arginine methyl ester)-induced HFpEF and metabolic syndrome in mice. We found that oral nitazoxanide improved cardiac hypertrophy, cardiac fibrosis, cardiac diastolic dysfunction, increased blood pressure, impaired exercise tolerance, impaired glucose handling, serum lipid disorders, hepatic steatosis, increased weight of white adipose tissues and kidney fibrosis in HFD + L-NAME-treated mice. In the established HFD + L-NAME-induced HFpEF and metabolic syndrome mouse model, therapeutic treatment with nitazoxanide rescued HFD + L-NAME-induced pathological phenotypes as mentioned above. The in vitro experiments revealed that tizoxanide, the active metabolite of nitazoxanide, increased the basal mitochondria metabolism of cardiomyocytes, inhibited cardiomyocyte hypertrophy and collagen secretion from cardiac fibroblasts, and relaxed phenylephrine- and U46619-induced constriction of rat mesenteric arteries, indicating that the direct effect of tizoxanide might partly contribute to the protective effect of nitazoxanide against HFpEF in vivo. The present study suggests that nitazoxanide might be a potential drug for HFpEF and metabolic syndrome therapy.

As a critical component of global agriculture,the dairy industry is essential for the food security and economic development.In China,the dairy sector in plateau regions plays an important role in addressing the nutritional needs of residents,boosting farmers' incomes,and fostering re-gional economic growth.However,the harsh high-altitude conditions,such as low oxygen,low at-mospheric pressure,and severe climate,pose significant challenges to the health and survival of dairy cows.This article summarizes the changes in physiological characteristics,metabolic perform-ance,production performance,and health status of dairy under high altitude areas,providing in-sights for the identification of adaptive traits in dairy to plateau.Also,the genetic and epigenetic mechanisms behind these phenotypic adaptations are discussed,and future directions and strategies for enhancing adaptability of dairy of dairy n plateau regions are outlined,thereby guiding re-searchers in adaptation evolution and breeding of dairy cattle.

Objective:To investigate the protective effect and its mechanism of proscillaridin A (PSD-A) on podocyte injury in lupus nephritis (LN).Methods:Molecular docking and surface plasmon resonance techniques were used to analyze the binding status of PSD-A to signal transducer and activator of transcription 1 (STAT1). The immortalized human podocyte injury model in the lupus group was induced by the serum of systemic lupus erythematosus patients, and the control and PSD-A intervention (2 nmol/L, 4 nmol/L) groups were also set up. Six female 12-week-old C57BL/6 mice were designated as the control group, and 12 female 12-week-old MRL/lpr lupus mice were randomly divided into lupus group and PSD-A intervention group by random number table method. The PSD-A intervention group was intraperitoneally injected with 5 mg/kg PSD-A, once per week for 6 consecutive weeks. While the control group and the lupus group were intraperitoneally injected with the same volume of the solvent without PSD-A. Western blotting and real-time quantitative PCR were employed to detect the relative protein and mRNA expression levels of podocin, STAT1, and interferon-induced protein with tetratricopeptide repeat 1 (IFIT1) in podocytes of each group. Enzyme-linked immunosorbent assay was used to detect the levels of serum anti-double strand DNA antibody and interferon-α in mice. Coomassie brilliant blue was used to detect the urinary protein level. HE, PAS, Masson and PASM staining and transmission electron microscopy were used to observe the pathological changes of renal tissues. Immunohistochemistry was used to examine the protein expression of podocin, STAT1 and IFIT1 in renal tissues.Results:Molecular docking and surface plasmon resonance techniques proved that PSD-A could bind to STAT1 protein and they exhibited a robust binding affinity. The podocyte experiments showed that, compared with the lupus group, the relative expression levels of podocin protein and mRNA in the PSD-A intervention group were upregulated, while the relative expression levels of STAT1 and IFIT1 protein and mRNA were downregulated (all P<0.05). The animal experiments showed that, compared with the lupus group, the serum levels of anti-double strand DNA antibody, interferon-α, and urinary protein in PSD-A intervention group were decreased, the pathological damage of renal tissues was alleviated, and the injury of renal podocytes was reduced. Immunohistochemical staining showed that the relative protein expression levels of STAT1 and IFIT1 of renal tissues in the PSD-A intervention group were lower than those in the lupus group (all P<0.05). Conclusion:PSD-A can play a protective role in podocyte injury in LN, and its mechanism may be related to the inhibition of the STAT1 signaling pathway.

Objective:To investigate the value of anlotinib combined with radiotherapy in second-line targeted drug-resistant advanced lung adenocarcinoma with epidermal growth factor receptor(EGFR) mutation.Methods:A total of 150 patients with advanced lung adenocarcinoma with EGFR mutation who were resistant to second-line targeted therapy and admitted to the Affiliated Hospital of Shaoxing University from July 2021 to July 2023 were prospectively selected as the study objects, and they were divided into the control group and the study group by random number table method, with 75 cases in each group. The control group received radiotherapy, and the study group received anrotinib combined radiotherapy. The recent clinical effects, tumor indexes, cellular immune indexes and the occurrence of adverse reactions were compared between the two groups.Results:After treatment, the objective response rate (ORR) and disease control rate (DCR) in the study group were higher than those in the control group: 53.33%(40/75) vs. 34.67%(26/75), 86.67%(65/75) vs. 64.00%(48/75), there were statistical differences ( χ2 = 5.30, 10.37, P<0.05). After treatment, the levels of VEGF, cytokeratin 21-1 fragment (CYFRA21-1) and carcinoembryonic antigen (CEA) in the study group were lower than those in the control group: (472.93 ± 45.71) ng/L vs. (510.26 ± 50.49) ng/L, (4.82 ± 1.13) μg/L vs. (5.25 ± 1.34) μg/L, (5.65 ± 1.24) μg/L vs. (6.17 ± 1.42) μg/L, there were statistical differences ( P<0.05). After treatment, the levels of CD 3+, CD 4+ and CD 4+/CD 8+ in the study group were higher than those in the control group, and the level of CD 8+ was lower than that in the control group: 0.532 ± 0.044 vs. 0.508 ± 0.041, 0.321 ± 0.030 vs. 0.305 ± 0.027, 1.02 ± 0.19 vs. 0.93 ± 0.16, 0.303 ± 0.040 vs. 0.320 ± 0.044, there were statistical differences ( P<0.05). There was no significant difference in the occurrence of adverse reactions between the two groups ( P>0.05). Conclusions:Anlotinib combined with radiotherapy can enhance the clinical efficacy of second-line targeted drug resistant advanced lung adenocarcinoma with EGFR mutation, improve tumor indexes, promote the recovery of immune function, without significantly increasing adverse reactions.

Objective To investigate the relationship between prolactin and mild cognitive impairment(MCI)in postmenopausal women with type 2 diabetes mellitus(T2DM).Methods A total of 319 postmenopausal women with T2DM who were hospitalized in the Department of Endocrinology,Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University were enrolled in this study from August 2016 to October 2023.All the patients were divided into two groups according whether they had MCI:T2DM group(n=161)and MCI group(n=158).Differences in clinical characteristics were compared between the two groups.Pearson correlation was used to analyze the correlation between sex hormones and cognitive domains,and Logistic regression analysis was used to evaluate the influencing factors for MCI development.Results Serum prolactin levels were significantly lower in the MCI group than in the T2DM group[(5.5±2.1)vs(7.2±2.9)μg/L,P<0.05].Serum prolactin level was positively correlated with mini-mental state examination score,Montreal cognitive assessment score,immediate memory score,visuopatial constructional score,attention score and hippocampal volume(P<0.05),and negatively correlated with processing speed test(time)and executive function test(time)(P<0.05).Logistic regression analysis demonstrated that serum prolactin level was an influencing factor for the risk of MCI in postmenopausal women with T2DM(OR 0.715,95%CI 0.605～0.845,P<0.01).Conclusions The decrease of serum prolactin level is associated with an increased risk of MCI in postmenopausal women with T2DM.

Objective To investigate the relationship between prolactin and mild cognitive impairment(MCI)in postmenopausal women with type 2 diabetes mellitus(T2DM).Methods A total of 319 postmenopausal women with T2DM who were hospitalized in the Department of Endocrinology,Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University were enrolled in this study from August 2016 to October 2023.All the patients were divided into two groups according whether they had MCI:T2DM group(n=161)and MCI group(n=158).Differences in clinical characteristics were compared between the two groups.Pearson correlation was used to analyze the correlation between sex hormones and cognitive domains,and Logistic regression analysis was used to evaluate the influencing factors for MCI development.Results Serum prolactin levels were significantly lower in the MCI group than in the T2DM group[(5.5±2.1)vs(7.2±2.9)μg/L,P<0.05].Serum prolactin level was positively correlated with mini-mental state examination score,Montreal cognitive assessment score,immediate memory score,visuopatial constructional score,attention score and hippocampal volume(P<0.05),and negatively correlated with processing speed test(time)and executive function test(time)(P<0.05).Logistic regression analysis demonstrated that serum prolactin level was an influencing factor for the risk of MCI in postmenopausal women with T2DM(OR 0.715,95%CI 0.605～0.845,P<0.01).Conclusions The decrease of serum prolactin level is associated with an increased risk of MCI in postmenopausal women with T2DM.