OBJECTIVE To screen and quantitatively analyze the quality marker （Q-Marker） associated with anti-renal fibrosis in Shenqi shenshuai mixture （SQSS）， so as to provide references for the analysis of pharmacodynamic substances and new drug transformation of SQSS. METHODS The chemical components of SQSS were characterized by untargeted metabolomics. Combined with network pharmacology， gene expression omnibus （GEO） and connectivity map （CMAP） databases， the anti-renal fibrosis Q-Markers of SQSS were screened. HPLC-MS/MS was applied to determine the contents of Q-Markers in 10 batches of SQSS. RESULTS A total of 1 319 compounds were identified from SQSS via untargeted metabolomics，and 84 active ingredients with anti-renal fibrosis activity were further screened out， such as formononetin. Nine anti-renal fibrosis Q-Markers were obtained by network pharmacology and GEO database， including berberine， quercetin， honokiol， nicotinamide， daidzein， coumarin， kaempferol， formononetin and amygdalin. The quantitative results showed that the average contents of the above components （excluding amygdalin） in 10 batches of SQSS were 2.523， 1.942， 26.848， 1.415， 0.692， 0.171， 0.374， 7.401 μg/mL， respectively. CONCLUSIONS Nine anti-renal fibrosis Q-Markers of SQSS were screened in this study， and the contents of eight among them were determined. The results can provide a basis for elucidating the pharmacodynamic material basis and promoting the new drug transformation of SQSS.

OBJECTIVE To screen and quantitatively analyze the quality marker （Q-Marker） associated with anti-renal fibrosis in Shenqi shenshuai mixture （SQSS）， so as to provide references for the analysis of pharmacodynamic substances and new drug transformation of SQSS. METHODS The chemical components of SQSS were characterized by untargeted metabolomics. Combined with network pharmacology， gene expression omnibus （GEO） and connectivity map （CMAP） databases， the anti-renal fibrosis Q-Markers of SQSS were screened. HPLC-MS/MS was applied to determine the contents of Q-Markers in 10 batches of SQSS. RESULTS A total of 1 319 compounds were identified from SQSS via untargeted metabolomics，and 84 active ingredients with anti-renal fibrosis activity were further screened out， such as formononetin. Nine anti-renal fibrosis Q-Markers were obtained by network pharmacology and GEO database， including berberine， quercetin， honokiol， nicotinamide， daidzein， coumarin， kaempferol， formononetin and amygdalin. The quantitative results showed that the average contents of the above components （excluding amygdalin） in 10 batches of SQSS were 2.523， 1.942， 26.848， 1.415， 0.692， 0.171， 0.374， 7.401 μg/mL， respectively. CONCLUSIONS Nine anti-renal fibrosis Q-Markers of SQSS were screened in this study， and the contents of eight among them were determined. The results can provide a basis for elucidating the pharmacodynamic material basis and promoting the new drug transformation of SQSS.

Osteoarthritis (OA) causes chronic pain that significantly impairs quality of life, with current treatments often proving insufficient and accompanied by adverse effects. Recent research has identified the dorsal root ganglion (DRG) and its resident macrophages as crucial mediators of chronic OA pain through neuroinflammation driven by macrophage polarization. We present a novel injectable thermo-sensitive hydrogel system, KAF@PLEL, designed to deliver an anti-inflammatory peptide (KAF) specifically to the DRG. This biodegradable hydrogel enables sustained KAF release, promoting the reprogramming of DRG macrophages from pro-inflammatory to anti-inflammatory phenotypes. Through comprehensive in vitro and in vivo studies, we evaluated the hydrogel's biocompatibility, effects on macrophage polarization, and therapeutic efficacy in chronic OA pain management. The system demonstrated significant capabilities in preserving macrophage mitochondrial function, suppressing neuroinflammation, alleviating chronic OA pain, reducing cartilage degradation, and improving motor function in OA rat models. The sustained-release properties of KAF@PLEL enabled prolonged therapeutic effects while minimizing systemic exposure and side effects. These findings suggest that KAF@PLEL represents a promising therapeutic approach for improving outcomes in OA patients through targeted, sustained treatment.

Objective To establish a human luminal breast cancer stem cell(BCSC)model and investigate the inhibitory effects of astragaloside Ⅳ(AS-Ⅳ)on BCSC growth.Methods MCF-7 breast cancer cells were cultured in stem cell-specific medium to induce BCSC formation.The BCSCs were then divided into a blank control group and an AS-Ⅳ treatment group,both groups were given PBS or AS-Ⅳ treatment.Morphological changes were observed after intervention.The therapeutic efficacy of AS-Ⅳ was evaluated using 3D spheroid formation and cell viability assays.Transcriptomic profiling and gene expression analysis were performed to elucidate the underlying mechanisms.Results Compared with the MCF7 breast cancer cells,MCF7 breast cancer stem cell mammospheres exhibited accelerated growth(P＜0.01)and significantly increased expression of the stemness marker ALDH1A1(P＜0.01).Further comparison with the blank control group revealed that astragaloside Ⅳ(AS-Ⅳ)treatment significantly inhibited MCF7 breast cancer stem cell proliferation(P＜0.001)and slowed mammosphere growth(P＜0.01).Transcriptomic analysis demonstrated that differentially expressed genes(DEGs)induced by stem cell modeling and AS-Ⅳ intervention were enriched in the cellular senescence signaling pathway.AS-Ⅳ intervention substantially increased the number of SA-β-gal-positive cells(P＜0.01).RT-PCR analysis confirmed that AS-Ⅳsignificantly upregulated mRNA expression of IL-1α(P＜0.01),P21(P＜0.001),and P53(P＜0.05)in MCF7 breast cancer stem cells.Conclusion Astragaloside Ⅳ suppresses the growth of human luminal breast cancer stem cells by inducing cellular senescence.

Acute postoperative pain and postoperative sleep disturbances are both major challenges in perioperative management,and they interact with each other.Acute pain can interfere with postoperative sleep,and sleep disturbances can lead to hyperalgesia and aggravate postoperative pain.At present,the in-teraction mechanism between the two is not clear,and there is also a lack of unified standards for prevention and control strategies.Therefore,this article reviews the research status of the definition,harmful effect,in-teraction mechanism,prevention,and management strategies of acute postoperative pain and postoperative sleep disturbances.We hope to provide valuable reference for the prevention and treatment of perioperative complications.

Objective To construct a 3D printed PLLA/β-tricalcium(PLLA/β-TCP)bone tissue engineering scaffold surface porous structure through simple treatment with NaOH solution,increase the roughness and hydrophilicity of the scaffold,and promote cell adhesion on the scaffold surface.Methods The PLLA/β-TCP mesh scaffold was prepared by 3D printing melt deposition molding technology,and the scaffold was roughed by NaOH etching.The effects of NaOH concentration and time on the scaffold were observed according to the microstructure,energy spectrum,contact angle,mechanics,and cell adhesion of the scaffold.Results The PLLA/β-TCP composite scaffold constructed by melt deposition technology had a pre-set porous structure,and the pores were interconnected.After NaOH etching,a porous structure with both macroscopic and microscopic pores was formed.The increase in any of the NaOH concentration and time parameters would lead to the increase of pore diameter and surface roughness.When the NaOH treatment parameter was 0.1 mol/L(9 h),it could significantly reduce the water contact angle on the surface of the scaffold,and had no significant effect on the compressive strength of the scaffold.In vitro cell testing showed that the surface porous composite scaffold etched with NaOH had more advantages in the adhesion and proliferation of BMSCs.Conclusion Using NaOH to process 3D printing of PLLA/β-TCP bone tissue engineering scaffolds can effectively improve the surface morphology of the scaffold,and optimize its hydrophilicity and cell adhesion.

Objective To solve the problem of insufficient hydrophilicity on the surface of polycaprolactone(PCL)/β-TCP bone tissue engineering scaffolds,NaOH etching method was used to improve the surface microstructure of 3D printed PCL/β-TCP scaffolds,further affecting their hydrophilicity and cell response.Methods PCL/β-TCP mesh scaffolds were prepared using 3D printing melt deposition molding technology,and the surface roughness of the scaffolds was modified by NaOH etching.The effects of two reaction parameters,NaOH concentration and time,on the microstructure,spectral elements,contact angle,compressive strength,and cell adhesion of the scaffolds before and after modification were observed.Results After NaOH etching,the surface microporous structure of the mesh scaffold was successfully prepared.With the increase of either NaOH concentration or time,the surface micropores of the scaffold increased while the contact angle of the material surface decreased.However,the compression strength of the etched scaffold treated with NaOH for 1 mol/L(24 h)or 10 mol/L(6 h)was not statistically significant compared to the untreated group(P＞0.05).The number of cells on the etched scaffold increased,with a larger spreading area of individual cells,making it more advantageous in the adhesion and proliferation of BMSCs.Conclusion The use of NaOH etching to improve the hydrophilicity of 3D printed PCL/β-TCP bone tissue engineering scaffolds is a low-cost and effective strategy which can effectively improve the wettability and cell adhesion of the scaffolds.

Objective:To develope a titanium specimen with good osteogenic activity through fabrication of a composite hydroxyapatite coating on ordered micro-/nanotextured titanium surface.Methods:An ordered micro-/nanotextured structure was prepared on the surface of titanium (the control), and then hydroxyapatite was deposited on the as-prepared ordered micro-/nanotextured structure by alternative loop immersion method. The ordered micro-/nanotextured structures before and after hydroxyapatite deposition were denoted as HA and MN, respectively. Surface morphology was observed using a scanning electron microscope. Bone marrow mesenchymal stem cells (BMMSC) were seeded on the surface of three different materials. Cell morphology was observed with a scanning electron microscope. Cell adhesion and cell proliferation were evaluated using 4', 6-diamidino-2-phenylindole staining and cell counting kit-8 assay, respectively. Extracellular matrix mineralization and the expression levels of osteogenesis-related genes were evaluated by alizarin red staining and real-time quantitative PCR, respectively. Each group has three samples in every experiment.Results:After alternative loop immersing, the MN's original microholes (20 μm in diameter) were retained, and the uniform petal-like hydroxyapatite was deposited on the MN's original titania nanotubes (70 nm in diameter). Compared with the control, BMMSC on MN and HA elongated further and intersected along the micron structure with noticeable pseudopodia and pseudoplates, and the trend was more pronounced especially on HA. The number of early adherent cells on HA was remarkably larger than that on the control and MN at each time point ( P<0.05). On day 1, the A value of cell proliferation on HA was significantly higher than that on the control and MN ( P<0.05). The A value of cell proliferation on HA was significantly lower than that on the control and MN on day 3 ( P<0.05). On day 7, the A value of cell proliferation on HA was significantly lower than that on MN ( P<0.05), but there was no statistically significant difference in the A value of cell proliferation between HA and the control on day 7 ( P>0.05). The Avalue of extracellular matrix mineralization on HA (0.607±0.011) was significantly higher than that on the control and MN (0.268±0.025 and 0.522±0.022, respectively) ( t=-0.25, P<0.001; t=-0.34, P<0.001). The expression levels of bone related genes on HA were significantly higher than those on the control and MN ( P<0.05). Conclusions:HA could promote the BMMSC adhesion and osteogenic differentiation, support BMMSC proliferation, and demonstrate good osteogenic activity.

Since the utilization of anthracyclines in cancer therapy, severe cardiotoxicity has become a major obstacle. The major challenge in treating cancer patients with anthracyclines is minimizing cardiotoxicity without compromising antitumor efficacy. Herein, histone deacetylase SIRT6 expression was reduced in plasma of patients treated with anthracyclines-based chemotherapy regimens. Furthermore, overexpression of SIRT6 alleviated doxorubicin-induced cytotoxicity in cardiomyocytes, and potentiated cytotoxicity of doxorubicin in multiple cancer cell lines. Moreover, SIRT6 overexpression ameliorated doxorubicin-induced cardiotoxicity and potentiated antitumor efficacy of doxorubicin in mice, suggesting that SIRT6 overexpression could be an adjunctive therapeutic strategy during doxorubicin treatment. Mechanistically, doxorubicin-impaired mitochondria led to decreased mitochondrial respiration and ATP production. And SIRT6 enhanced mitochondrial biogenesis and mitophagy by deacetylating and inhibiting Sgk1. Thus, SIRT6 overexpression coordinated metabolic remodeling from glycolysis to mitochondrial respiration during doxorubicin treatment, which was more conducive to cardiomyocyte metabolism, thus protecting cardiomyocytes but not cancer cells against doxorubicin-induced energy deficiency. In addition, ellagic acid, a natural compound that activates SIRT6, alleviated doxorubicin-induced cardiotoxicity and enhanced doxorubicin-mediated tumor regression in tumor-bearing mice. These findings provide a preclinical rationale for preventing cardiotoxicity by activating SIRT6 in cancer patients undergoing chemotherapy, but also advancing the understanding of the crucial role of SIRT6 in mitochondrial homeostasis.

Objective To investigates the effect of acupuncture on sensitization of Zusanli(ST36)in rats with different functional states by using healthy and knee osteoarthritis model rats.Methods Male SD rats were randomly divided into control,model,model-acupuncture and blank-acupuncture group,with 7 rats in each group.KOA rat model was prepared by intra-articular injection of 1 mg·50 μL-1 monoiodoacetic aci(MIA)in model group and model-acupuncture group.On the second day of modeling,acupuncture treatment was performed on the left Zusanli of the model acupuncture group and the blank-acupuncture group,once everyday for 20 min,5 times as a course of treatment,2 days between courses.The general condition,knee joint diameter,plantar thermal pain threshold and Lequesne MG score of rats was observed before modeling and after acupuncture.Observing the morphology of knee joint cartilage to judge whether the model is successful,measuring the mechanical pain threshold of Zusanli to investigate the acupoint sensitization,observing and counting the morphology of skin mast cells in the acupoint area,and detecting the expression of skin calcitonin gene-related peptide(CGRP)in the acupoint area.Results The mechanical pain threshold of Zusanli after acupuncture in model group and blank-acupuncture group decreased significantly after modeling(P<0.01,P<0.05),compared with the control group,the change rate of mechanical pain threshold in model group and blank-acupuncture group increased significantly(P<0.05),compared with the model group,the mechanical pain threshold of Zusanli in the model-acupuncture group decreased significantly(P<0.01).Compared with the control group,the fluorescence intensity of CGRP protein in the skin tissue of Zusanli in the model group increased significantly(P<0.01),MC degranulation rate increased significantly(P<0.05),and there was no significant difference in the fluorescence intensity of CGRP protein of Zusanli in the blank-acupuncture group(P>0.05),MC degranulation rate increased obviously(P<0.01),CGRP protein of Zusanli in the model-acupuncture group was significantly reduced compared with the model group(P<0.01),and there was no significant difference in the degranulation rate of MC(P>0.05).Conclusion Acupoint sensitization can occur in different functional states of rats.Zusanli(ST36)of KOA model rats can be sensitized,and acupuncture stimulation can make Zusanli sensitization caused by disease disappear.Under physiological conditions,acupuncture stimulation can induce similar sensitization phenomenon.