Based on spleen deficiency-phlegm dampness as the core pathogenesis of obesity， and integrating recent advances in modern medicine regarding the key role of immune inflammation in obesity， this paper proposes a multidimensional pathogenic network of "obesity-spleen deficiency-phlegm dampness-immune imbalance". Various traditional Chinese medicine （TCM） herbs that strengthen the spleen， regulate qi， and resolve phlegm and dampness can treat obesity by improving spleen-stomach transport and transformation， promoting water-damp metabolism， and regulating immune homeostasis. This highlights immune inflammation as an important entry point to elucidate the TCM concepts of "spleen deficiency-phlegm dampness" and the therapeutic principle of "strengthening the spleen and eliminating dampness to treat obesity". By systematically analyzing the intrinsic connection between "spleen deficiency generating dampness， internal accumulation of phlegm dampness" and immune dysregulation in obesity， this paper aims to provide theoretical support for TCM treatment of obesity based on dampness.

Background: Malignant pleural effusion (MPE), a common complication of advanced cancers, is associated with poor prognosis and reduced quality of life. Although host–tumor interactions are known to drive MPE development, the associated immune dynamics during disease progression remain unclear. Using a Lewis lung carcinoma-induced MPE model in C57BL/6JNidfc mice, we systematically evaluated general parameters and immune cell changes at two-day intervals throughout disease progression. Results: The day of Lewis lung carcinoma cell injection into the pleural space was designated as day 0. By day 10 post-injection (p.i.), MPE-bearing mice exhibited ~ 10% body weight loss, marking the experimental endpoint. Pleural tumor mass and pleural effusion volume were minimal up to day 4 p.i. but increased sharply from day 6 onward.CD45⁺ immune cell counts rose over time, and days 6, 8, and 10 p.i. marked key stages of MPE progression. On day 6, B cells, T cells, and natural killer cells, but not macrophages and neutrophils, increased significantly compared to earlier timepoints. By day 8, all immune cell subsets except T cells exceeded day 6 levels, and at day 10, natural killer cell numbers declined while others continued to increase. Besides, the numbers of CD8⁺ T cells, Th1 cells, regulatory T cells, and M2 macrophages progressively increased from day 6 to 10. Based on these data, days 6 and 10 were defined as early and advanced MPE stages, respectively, with distinct immune phenotypes. In advanced MPE, CD8⁺ T cells displayed reduced IFN-γ, TNF-α, Granzyme B, Perforin, FasL, and Ki-67, but upregulated PD-1 and CTLA-4 relative to early stage. Similarly, Th1 cells showed decreased IFN-γ, TNF-α, and IL-2 production along with reduced Ki-67 expression. Advanced-stage M2 macrophages exhibited lower MHC-II levels and impaired phagocytosis, but higher PD-L1 and IL-10 production, while neutrophils showed reduced TNF-α release and phagocytic activity. Conclusions Our findings characterize the temporal immune dynamics associated with MPE progression in a mouse model, revealing a transition from an early immunostimulatory state to a late immunosuppressive state. This study enhances our understanding of MPE immunopathogenesis and provides a foundation for developing precise, stagespecific therapeutic strategies.

The objective of this study was to examine the effects of tributyrin(TB)on energy me-tabolism,oxidative stress and immune function in dairy cows with negative energy balance(NEB).In this study,25 perinatal Holstein cows from a dairy farm in Heilongjiang Province were selected.Five healthy cows were designated as the control group,and 20 cows with NEB were randomly di-vided into NEB control group,Propylene glycol(PG)group(300 mL PG),TB group(37.5 g TB)and PG+TB group(300 mL PG+37.5 g TB)with 5 cows in each group,and were continuously fed for 14 days.Blood samples were collected at 0,7,14 and 21 days.The indexes of oxidative stress,energy metabolism and immune function in the serum of dairy cows were quantified using commercially available kits.The results showed that compared with the NEB group,the contents of T-AOC,GSH-Px,CAT and SOD in the TB group were significantly increased,whereas the content of MDA was significantly decreased.These results indicated that TB can effectively alleviate the oxidative stress of cows with NEB.The contents of BHB,NEFA and BUN in the TB group were significantly decreased,indicating that TB can regulate energy metabolism disorders in cows with NEB.The contents of IL1β,IL-2,IL-6,IL-21,MIP-1 and IP-10 in the TB group were significantly decreased,while the contents of IgA,IgM and IgG were significantly increased.These results sug-gest that TB has shown significant effects in inhibiting inflammatory response and improving im-mune function.Furthermore,compared with the PG group,the contents of T-AOC,GSH-Px,CAT,SOD,IgA,IgM and IgG in the TB group and the PG+TB group were significantly in-creased,whereas the contents of MDA,BHB,NEFA,BUN,IL-1β,IL-2,IL-6,IL-21,MIP-1 and IP-10 were significantly decreased.These results indicated that feeding TB alone was effective in alle-viating oxidative stress,improving energy metabolism and inhibiting inflammation in dairy cows compared to feeding the traditional drug PG,and the effect of feeding a mixture of PG and TB was particularly significant.This study provides a new theoretical basis for the prevention and treat-ment of oxidative stress and immune function decline caused by perinatal cows with NEB.

The aim of this study is to investigate whether β-hydroxybutyrate(BHB)impaired the mitochondrial function of dairy cow monocytes through the peroxisome proliferator-activated re-ceptor γ coactivator 1 alpha(PGC-1α)signaling pathway.According to the clinical symptoms and the concentration of BHB in whole blood,the tail vein blood of 12 healthy cows(BHB＜1.2 mmol/L)and 12 clinical ketotic cows(CK,BHB＞3.0 mmol/L)was collected.In vivo,after isolation and purification of CD14+monocytes,the intracellular mitochondrial membrane potential was detected by flow cytometry.The protein abundance of oxidative phosphorylation complex cytochrome c oxi-dase subunit Ⅰ(CO Ⅰ),CO Ⅱ,CO Ⅲ,COⅣ,CO Ⅴ,PGC-1 α,mitochondrial transcription factor A(TFAM)and nuclear respiratory factor 1(NFR1)were determined by Western blot.In vitro,CD14+monocytes were co-cultured with 3.0 mmol/L BHB for 0,6,12,24 h.Flow cytometry was applied for intracellular mitochondrial membrane potential detection,and determination the protein abundance of PGC-1α,TFAM and NFR1 by Western blot.The results showed that compared with control group,the mitochondrial membrane potential in CD14+monocytes of ketotic cows was sig-nificantly increased,and the protein abundance of CO Ⅰ,CO Ⅱ,CO Ⅲ,CO Ⅳ,CO Ⅴ PGC-1α,TFAM,and NRF1 in CD14+monocytes of ketotic cows were significantly decreased.Compared with 0 h,the mitochondrial membrane potential of CD14+monocytes was significantly increased after BHB treatment for 6,12,24 h,and the protein abundance of PGC-1α,TFAM and NRF1 were significantly decreased.The results indicated that BHB induced mitochondrial dysfunction in CD14+monocytes of ketotic cows by inhibiting PGC-1α signaling pathway.Therefore,the PGC-1αsignaling pathway may be a preventive and therapeutic target to the mitochondrial dysfunction of monocytes in ketotic cows caused by BHB.

To clarify the current situation of ketosis in dairy cattle on large-scale pastures in China and provide new insights,a questionnaire survey was conducted to analyze the incidence,preven-tion,treatment methods,and associated costs of ketosis in 79 large-scale pastures.The results showed that the average incidence of ketosis in dairy cows was 3.97％,with a cure rate of 92.40％.The order of importance of methods for preventing and controlling ketosis was as follows:feed for-mulation optimization＞blood ketone monitoring＞negative energy balance monitoring＞feed in-take monitoring＞milk yield monitoring.The most important treatment methods are intravenous glucose＞propylene glycol butyl phosphate＞vitamins＞choline.The most important diagnostic methods are blood ketone testing＞milk ketone testing＞negative energy balance testing＞clinical symptoms＞blood glucose testing.Economic analysis revealed that treatment costs were lower on larger farms and higher milk yields farms.Continuous optimization of feeding management,preven-tion,and control measures should be implemented on large-scale farms in China to reduce the oc-currence of ketosis in dairy cows.Additionally,more effective diagnostic and treatment methods should be employed to improve the cure rate and overall farm income.

In recent years,the intensification of dairy farming has significantly improved production efficiency but has also led to a rise in the incidence of metabolic diseases.Conditions such as keto-sis,fatty liver,and hypocalcemia pose serious threats to dairy cattle health and productivity.These diseases not only profoundly affect individual physiological function but also impose considerable economic pressures and challenges on farm management.As research advances,exosomes have e-merged as a novel intercellular signaling molecule,showing unique potential in regulating dairy cattle's nutritional metabolism.Studies suggest that exosomes hold promise as biomarkers for dis-ease and can even serve as carriers for disease detection and prevention.Acting as a crucial mediator of intercellular communication,exosomes play an important role in modulating the metabolic processes of dairy cattle.This review aims to systematically explore the role of exosomes in bovine nutritional metabolism and to provide new perspectives and theoretical support for their potential as tools for diagnosing,treating,and preventing metabolic diseases in dairy cattle,thus advancing research and practice in this field.

To investigate the effect of VD3 on the concentration of Ca2+in neutrophils of subclinical hypocalcemia dairy cows and the autophagy of LC3,peripheral blood of dairy cows was collected and neutrophils were isolated and extracted.Western blot was used to detect the abundance of LC3 autophagy pathway-related proteins(LC3-Ⅱ,P62,ATG5)in cells.The intracellular Ca2+concen-tration of neutrophils was detected by flow cytometry.The LC3 autophagosome was detected by la-ser confocal technology.By adding VD3 to affect intracellular Ca2+concentration levels,the effects of vitamin D3 on calcium absorption and LC3 autophagy function were investigated.Western blot result showed that the protein abundances of LC3 and ATG5 in neutrophils of subclinical hypocal-cemia dairy cows decreased,while the protein abundance of p62 increased,indicating that the auto-phagy function of LC3 in subclinical hypocalcemia dairy cows was weakened.After VD3 treatment,the flow cytometry results showed that VD3 promoted the increase of Ca2+concentration levels in neutrophils.The Western blot results showed that the LC3 autophagy function of neutrophils from healthy cows and subclinical hypocalcemia cows was enhanced under the action of VD3,indicating that VD3 promoted the concentration of Ca2+in neutrophils and then affected the LC3 autophagy function.

To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.

The peripheral blood of healthy or ketosis dairy cows was collected,and CD4+T cells were isolated.The expressions of lipid synthesis related proteins fatty acid synthase(FASN),acetyl coenzyme A carboxylase 1(ACC1),cluster of differentiation 36(CD36)and store-operated calcium entry(SOCE)related proteins ORAIl,ORAI2,ORAI3,STIM1,STIM2 were detected by Western blot.IL-17 cells were detected by flow cytometry.CD4+T cells were isolated from the spleen of 1-day-old calves and cultured in vitro.Cells were treated and divided into control(Ctrl)group,si-lenced CD36(siCD36)group,stearic acid(SA)group,and SA+siCD36 group.Cells in the Ctrl and SA groups were transfected with 75 pmol/L negative control siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h;Cells in the siCD36 group and SA+siCD36 group were transfected with 75 pmol/L CD36 siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h in the SA+siCD36 group.The protein expression of FASN,CD36,ACC1,ORAI1,ORAI2,ORAI3,STIM1 and STIM2 was detected by Western blot,and IL-17 cells were detected by flow cytometry.The results showed that the expression of IL-17 in peripheral blood CD4+T cells of ketosis dairy cows was significantly increased compared to that of healthy cows(P＜0.01).Additionally,the protein level of FASN,CD36,STIM1(P＜0.05),and ACC1,ORAI2,ORAI3,STIM2(P＜0.01)were up-regulated.Compared with the Ctrl group,the protein expression levels of CD36,ACC1 and ORAI3(P＜0.05)were up-regulated in the SA group,as well as the protein expression of FASN and STIM1(P＜0.01).Additionally,the expression of IL-17 was significantly increased(P＜0.05).Compared with the SA group,there was a decrease in the protein expression of STIM1,ORAI1(P＜0.05)and CD36,ACC1,FASN,ORAI2(P＜0.01)in the siCD36+SA group,as well as IL-17(P＜0.05).These results suggest that SA can promote the expression of IL-17 in CD4+T cells in ketosis cows by regulating fatty acid synthesis and activating SOCE channels through CD36.

This study aims to clarify the incidence and prevention status of dairy cow mastitis in 79 large-scale pastures in China in 2022,reduce the incidence of dairy cow mastitis,and provide scien-tific basis for formulating prevention and control strategies for dairy cow mastitis in large-scale pastures suitable for China's national conditions.The research team relied on the large-scale ranch of the Comprehensive Experimental Station of National Dairy Industry Technology System to car-ry out research.A total of 79 questionnaires were received.The information on the stock of large-scale farms,the proportion of cows in the herd,the level of cow yields,the incidence and cure rate of cow mastitis,the prevention and control of cow mastitis,diagnosis and treatment programs of cow mastitis,the cost of treating cow mastitis,and the culling rate of farms was collected.Chi-square test and correlation analysis were carried out on the incidence of mastitis in dairy cows and the prevention and control plan,treatment plan,yield level and elimination rate of mastitis in dairy cows by descriptive statistical cross table.The incidence of cow mastitis in some pastures was con-trolled between 5％and 10％,and the cure rate was mostly as high as 98％.According to the co-hort analysis,the importance of prevention and control measures for dairy cow mastitis was as fol-lows:DMT＞DHI＞milk yield＞milk traits＞breast apparent change＞electrical conductivity＞CMT.The importance of treatment measures for dairy cow mastitis is as follows:DMT＞DHI＞milk yield＞milk traits＞breast apparent changes＞electrical conductivity＞CMT.The importance of treatment measures for mastitis is as follows:traditional Chinese medicine＞pathogen detection and drug sensitivity test＞non-steroidal anti-inflammatory drugs＞regular maintenance of milking equipment＞normal milking without treatment＞broad-spectrum antibiotics.The incidence of mas-titis in dairy cows is moderately positively correlated with the yield per unit area,that is,the in-crease in the incidence of mastitis in dairy cows has a certain impact on the yield per unit area of dairy cows.Economic analysis shows that the treatment cost of mastitis is closely related to the in-cidence rate,and the treatment cost of pasture with higher incidence rate is higher.Pastures with higher yields usually have more efficient means of disease prevention and control,and the corre-sponding treatment costs are lower.In conclusion,China's large-scale pastures should strengthen the scientific feeding management level,improve the prevention and control methods to reduce the incidence of cow mastitis,improve the cure rate,and reduce the cost of pasture treatment.