ObjectiveThis paper aims to conduct a secondary analysis of a randomized controlled trial on the treatment of myelodysplastic syndrome （MDS） with deficiency of both the spleen and kidney and blockage of toxin and blood stasis with an arsenic-containing traditional Chinese medicine compound， by applying the mixed model for repeated measure （MMRM） and the method of stratified composite outcome with win ratio. The analysis includes the assessment of hematological efficacy and the composite outcome evaluation of adverse reactions， so as to more comprehensively assess the therapy of this regimen. MethodsThe MMRM and win ratio methods were used to evaluate the efficacy of a prospective，multi-center，double-blind，randomized controlled study. The blood routine （hemoglobin concentration，neutrophil count， and platelet count） and biochemical indexes （aspartate aminotransferase，alanine aminotransferase，serum creatinine，and serum ferritin） of the patients were detected at the time of enrollment and at the end of each course of treatment in the laboratory department of Xiyuan Hospital. The patients' syndromes at the time of enrollment and after treatment were recorded and scored according to the therapy standard of traditional Chinese medicine for diseases and syndromes. MMRM was used to analyze the blood routine indexes of the experimental group and the control group. This method has the advantages of high data reliability and dynamic efficacy under intervention and time. The win ratio method was used to evaluate the composite outcome of traditional Chinese medicine syndrome scores and biochemical indexes according to the priority and to verify the clinical safety of arsenic-containing traditional Chinese medicine compound. ResultsThe results of MMRM analysis showed that the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly compared with that before treatment in the group，while that in the placebo group decreased significantly （P<0.01）. When compared with that after treatment in the placebo group，the hemoglobin concentration of patients in the group with arsenic-containing traditional Chinese medicine compound increased significantly，and the mean difference of least squares （LS） was statistically significant （P<0.01）. When compared with those before treatment in the group，there were no statistically significant differences in the neutrophil count and platelet count in both groups. After treatment，there were no statistically significant differences in the neutrophil count， platelet count， and the mean difference of LS between the two groups. The analysis results of win ratio showed that the group with arsenic-containing traditional Chinese medicine compound had a significant advantage in the comparison of composite outcomes，with a win ratio （95% CI） of 2.01 （1.24-3.27） （P<0.01），and that the possibility of "winning" in terms of safety was 2.01 times that of the placebo group. The safety advantage of the group with arsenic-containing traditional Chinese medicine compound mainly came from the traditional Chinese medicine syndrome scores，renal function indexes， and iron reserve capacity indexes，and the number of winning times was less than that of losing times in the comparison of liver function outcomes. ConclusionThe MMRM analysis proves that the arsenic-containing traditional Chinese medicine compound can significantly improve the hemoglobin concentration of patients with myelodysplastic syndrome with refractory cytopenia and multilineage dysplasia （MDS-RCMD） of the type of deficiency of both the spleen and kidney and blockage of toxin and blood stasis. This conclusion is not interfered with by time trends and individual relationships and methodologically improves the credibility of the therapy of the arsenic-containing traditional Chinese medicine compound in treating MDS. Four outcomes are evaluated by the win ratio method，namely traditional Chinese medicine syndromes，liver function，renal function， and iron reserve capacity，proving that the arsenic-containing traditional Chinese medicine compound has the comprehensive advantages of improving the survival quality of the patients and reducing adverse reactions. The win ratio outcome provides clear comparative indexes for the evaluation of adverse reactions，making it easier for regulatory authorities，medical staff， and patients to understand the safety of the arsenic-containing traditional Chinese medicine compound in clinical application.

The aryl hydrocarbon receptor (AhR) plays a crucial role in regulating many physiological processes. Activating the AhR-CYP1A1 axis has emerged as a novel therapeutic strategy against various inflammatory diseases. Here, a practical in situ cell-based fluorometric assay was constructed to screen AhR-CYP1A1 axis modulators, via functional sensing of CYP1A1 activities in live cells. Firstly, a cell-permeable, isoform-specific enzyme-activable fluorogenic substrate for CYP1A1 was rationally constructed for in-situ visualizing the dynamic changes of CYP1A1 function in living systems, which was subsequently used for discovering the efficacious modulators of the AhR-CYP1A1 axis. Following screening of a compound library, LAC-7 was identified as an efficacious activator of the AhR-CYP1A1 axis, which dose-dependently up-regulated the expression levels of both CYP1A1 and AhR in multiple cell lines. LAC-7 also suppressed macrophage M1 polarization and reduced the levels of inflammatory factors in LPS-induced bone marrow-derived macrophages. Animal tests showed that LAC-7 could significantly mitigate DSS-induced ulcerative colitis and LPS-induced acute lung injury in mice, and markedly reduced the levels of multiple inflammatory factors. Collectively, an optimized fluorometric cell-based assay was devised for in situ functional imaging of CYP1A1 activities in living systems, which strongly facilitated the discovery of efficacious modulators of the AhR-CYP1A1 axis as novel anti-inflammatory agents.

Notum, a negative feedback regulator of the Wnt signaling, has emerged as a promising target for treating glucocorticoid-induced osteoporosis (GIOP). This study showcases an efficient strategy for discovering the anti-Notum constituents from herbal medicines (HMs) as novel anti-GIOP agents. Firstly, a rapid-responding near-infrared fluorogenic substrate for Notum was rationally engineered for high-throughput identifying the anti-Notum HMs. The results showed that Bu-Gu-Zhi (BGZ), a known anti-osteoporosis herb, potently inhibited Notum in a competitive-inhibition manner. To uncover the key anti-Notum constituents in BGZ, an efficient strategy was adapted via integrating biochemical, phytochemical, computational, and pharmacological assays. Among all identified BGZ constituents, three furanocoumarins were validated as strong Notum inhibitors, while 5-methoxypsoralen (5-MP) showed the most potent anti-Notum activity and favorable safety profiles. Mechanistically, 5-MP acted as a competitive inhibitor of Notum via creating strong hydrophobic interactions with Trp128 and Phe268 in the catalytic cavity of Notum. Cellular assays showed that 5-MP remarkably promoted osteoblast differentiation and activated Wnt signaling in dexamethasone (DXMS)-challenged MC3T3-E1 osteoblasts. In dexamethasone-induced osteoporotic mice, 5-MP strongly elevated bone mineral density (BMD) and improved cancellous and cortical bone thickness. Collectively, this study constructs a high-efficient platform for discovering key anti-Notum constituents from HMs, while 5-MP emerges as a promising anti-GIOP agent.

Triple-negative breast cancer is therapeutically challenging due to the low expression of tumor markers and 'cold' tumor immunosuppressive microenvironment. Here, we present a dual-targeting peptide-drug conjugate (PDC) for tumor inhibition. Our PDC efficiently and selectively delivers cytotoxic Monomethyl Auristatin E (MMAE) into tumor cells via C-X-C chemokine receptor type 4 (CXCR4) and folate receptor 1 (FOLR1) for synergistic inhibition of growth and metastasis. Our results show that the dual-targeting PDC has potent antitumor activity in cultured human cells and several murine transplanted tumor models without apparent toxicity. The combination of dual-targeting PDC and radiotherapy modulates the tumor immunosuppressive microenvironment by increasing CD8⁺ T cell infiltration and attenuating the proportion of myeloid-derived suppressor and regulatory T cells. Therefore, our dual-targeting PDC represents a promising new strategy for cancer therapy that rebalances the immune system and promotes tumor regression.

Objective To investigate the characteristics of peripheral blood in the treatment of aplastic anemia(AA)with Bushen Shengxue Method and Yiqi Yangxue Method combined with Western medicine.Methods Totally 492 AA patients who were treated in 19 centers including Xiyuan Hospital China Academy of Chinese Medical Sciences from September 2018 to March 2021 were selected,and were randomized into three groups:161 cases in the Bushen Shengxue group,164 cases in the Yiqi Yangxue group,and 167 cases in the control group.All three groups were orally administered cyclosporine and androgens.Bushen Shengxue group was given Bushen Shengxue Granules,Yiqi Yangxue group was given Yiqi Yangxue Granules,and the control group was given placebo(half dosage of Bushen Shengxue granules).The general data,overall efficacy,and peripheral blood at the 1st month,the 4th month,and 6th month after treatment were analyzed.Results The total effective rate of Bushen Shengxue group was 98.8％(159/161),which was significantly higher than that of Yiqi Yangxue group(79.9％)and the control group(61.7％),with statistical significance(P＜0.001);The total effective rate of Yiqi Yangxue group was significantly higher than that of the control group(P＜0.001).Compared with before treatment,the hemoglobin(HGB)levels of all three groups of patients significantly increased after treatment(P＜0.001,P＜0.01),and the platelet levels of Bushen Shengxue group and the control group significantly increased after treatment(P＜0.001);after treatment,the HGB levels in the three groups were ranked from high to low as the Bushen Shengxue group,Yiqi Yangxue group and the control group(P＜0.01).25％of patients had HGB levels exceeding 20％of baseline values,Bushen Shengxue group took 1 month,while Yiqi Yangxue group and control group took 4 months;25％of patients had an increase in HGB levels exceeding 50％of the baseline value,Bushen Shengxue group took 6 months,while Yiqi Yangxue group and control group took more than 6 months.At 6 months,the number of patients with HGB elevation exceeding 20％and 50％of baseline values in Bushen Shengxue group was higher than that in Yiqi Yangxue group and the control group(P＜0.05,P＜0.001).25％of patients had white blood cell elevation exceeding 50％of the baseline value,Bushen Shengxue group took 4 months,Yiqi Yangxue group took 6 months,and the control group took more than 6 months.25％of patients showed an increase in platelet levels exceeding 100％of the baseline value,Bushen Shengxue group took 4 months,while Yiqi Yangxue group and control group took 6 months;at 6 months,the number of patients in Bushen Shengxue group and Yiqi Yangxue group with platelet elevation exceeding 20％of the baseline value was significantly higher than that in the control group(P＜0.01,P＜0.05).Conclusion Bushen Shengxue method combined with Western medicine intreating AA is better than the method of Yiqi Yangxue method combined with Western medicine.In terms of improving HGB,it is reflected in shortening the recovery time and increasing the number of beneficiaries,and the dosage can affect the recovery time and the number of beneficiaries at the same time;in terms of improving white blood cell and platelet,it is reflected in shortening the recovery time,and the dosage can affect the recovery time.

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.

Signaling lymphocyte activation molecule family 6(SLAMF6)is widely expressed in T,B lymphocytes,NK cells and other immune cells.As a self-binding receptor for homotypic interactions,SLAMF6 is a type Ⅰ transmembrane protein belonging to the immunoglobulin superfamily,composed of the proximal IgC2 domain and distal IgV domain,the extracellular membrane,and three unique tyrosine cytoplasmic signaling motifs.This article summarizes the structure,expression,function and implication for au-toimmunity and tumor immunity of SLAMF6.

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.