This study aims to explore the scientific connotation of sinking Rehmanniae Radix has the best quality and compare the quality between floating and sinking fresh Rehmanniae Radix samples. Ultra-performance liquid chromatography tandem quadrupole electrostatic field Orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was employed to detect the chemical components in floating and sinking fresh Rehmanniae Radix samples. The fingerprint of fresh Rehmanniae Radix was established by high performance liquid chromatography(HPLC), and four index components were determined simultaneously. The cluster analysis, principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) were conducted to compare the quality of floating and sinking fresh Rehmanniae Radix samples. An evaporative light-scattering detector was used to compare the content of five sugars. The extract yield and drying rate were determined, and the quality connotation of sinking Rehmanniae Radix has the best quality was explained by multiple indicators. A total of 41 components were preliminarily identified from fresh Rehmanniae Radix by UHPLC-Q-Orbitrap HRMS, including 7 iridoid glycosides, 9 phenylethanol glycosides, 6 amino acids, 4 sugars, 3 phenolic acids, 5 nucleosides, 3 organic acids, 1 ionone, 1 furan, 1 coumarin, and 1 phenylpropanoid. The results showed that the main chemical components were consistent between floating and sinking fresh Rehmanniae Radix. Nine common peaks were identified in the fingerprints of 15 batches of floating and sinking fresh Rehmanniae Radix samples, and the similarity of fingerprints was greater than 0.9. The cluster analysis, PCA, and OPLS-DA classified floating and sinking fresh Rehmanniae Radix sasmples into two categories, indicating differences in the quality between them. The total content of catalpol, rehmannioside D, ajugol, and verbascoside in sinking fresh Rehmanniae Radix samples was higher than that in floating samples of the same batch and specification, and the main differential component was catalpol. The total content of fructose, glucose, sucrose, raffinose, and stachyose in sinking fresh Rehmanniae Radix samples was higher than that in floating samples of the same batch and specification, and the main differential component was stachyose. The extract yield and drying rate of the sinking samples were higher than those of floating samples. This study preliminarily showed that floating and sinking fresh Rehmanniae Radix samples had the same components but great differences in the content of medicinal substance basis. The total content of four glycosides and five sugars, extract yield, and drying rate of sinking fresh Rehmanniae Radix samples is higher than that of floating samples of the same batch and specification. These findings, to a certain extent, explains the scientificity of sinking Rehmanniae Radix has the best quality recorded in ancient books and provide a reference for the quality control and clinical application of fresh Rehmanniae Radix.
Chromatography, High Pressure Liquid/methods* ; Drugs, Chinese Herbal/chemistry* ; Rehmannia/chemistry* ; Chemometrics ; Mass Spectrometry/methods* ; Quality Control ; Principal Component Analysis ; Plant Extracts

Radiation-induced thrombocytopenia (RIT) faces a perplexing challenge in the clinical treatment of cancer patients, and current therapeutic approaches are inadequate in the clinical settings. In this research, oxymatrine, a new molecule capable of healing RIT was screened out, and the underlying regulatory mechanism associated with magakaryocyte (MK) differentiation and thrombopoiesis was demonstrated. The capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro. The ability to induce thrombopoiesis was subsequently demonstrated in Tg (cd41:enhanced green fluorescent protein (eGFP)) zebrafish and RIT model mice. In addition, we carried out network pharmacological prediction, drug affinity responsive target stability assay (DARTS) and cellular thermal shift assay (CETSA) analyses to explore the potential targets of oxymatrine. Moreover, the pathway underlying the effects of oxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, Western blot (WB), and immunofluorescence. Oxymatrine markedly promoted MK differentiation and maturation in vitro. Moreover, oxymatrine induced thrombopoiesis in Tg (cd41:eGFP) zebrafish and accelerated thrombopoiesis and platelet function recovery in RIT model mice. Mechanistically, oxymatrine directly binds to toll-like receptor 2 (TLR2) and further regulates the downstream pathway stimulator of interferon genes (STING)/nuclear factor-kappaB (NF-κB), which can be blocked by C29 and C-176, which are specific inhibitors of TLR2 and STING, respectively. Taken together, we demonstrated that oxymatrine, a novel TLR2 agonist, plays a critical role in accelerating MK differentiation and thrombopoiesis via the STING/NF-κB axis, suggesting that oxymatrine is a promising candidate for RIT therapy.

Aim To investigate the protective effect of Shengmai Yin on myocardial ischemia/reperfusion in-jury(MI/RI)in vitro and in vivo and to unravel the underlying mechanism.Methods SD rats were divid-ed into the sham group,model group,and Shengmai Yin group(SM).Rat MI/RI model was established.Cardiac function,infarct area,pathological changes,cardiomyocyte apoptosis,macrophage infiltration,and serum cTnT and CK-MB levels were measured.The mRNA and protein expressions of Calpain-1 and Cal-pain-2 were assessed.The hypoxia/reoxygenation(H/R)model was constructed in H9c2 cells.The active ingredients of Shengmai Yin were screened using net-work pharmacology and verified by CCK-8.In the car-diomyocytes H/R model,Fluo-4 AM staining was used to detect the changes of Ca2+levels.Results Com-pared with model group,LVEF and LVFS of Shengmai Yin-treated rats increased,myocardial infarction area was reduced,while myocardial tissue injury was allevi-ated.Myocardial apoptosis rate and the number of macrophages were reduced.Similarly,cTnT and CK-MB levels decreased.In addition,the expression lev-els of Calpain-1 and Calpain-2 mRNA and protein de-creased in the SM treatment group.Under the H/R model,all the active ingredients of Shengmai decoction had protective effects on cardiomyocytes,and the treat-ment could reduce the level of Ca2+in cardiomyocytes.Conclusions Shengmai Yin has protective effects on MI/RI in rats.This effect may be related to the de-crease in Ca2+levels,as well as Calpain-1 and Calap-in-2 mRNA and protein expression.

BACKGROUND:There are few studies on the effect of different patellar morphologies on the outcome after unicompartmental knee arthroplasty.OBJECTIVE:To investigate the effect of different patellar morphologies on functional recovery and patellofemoral joint alignment after unicompartmental knee arthroplasty based on patellar Wiberg classification.METHODS:A retrospective study was conducted in 186 patients with medial knee osteoarthritis who underwent unicompartmental knee arthroplasty at Affiliated Hospital of Xuzhou Medical University between January 2022 and March 2023.Patients were categorized into group A(type Ⅰ,n=43),group B(typeⅡ,n=104),and group C(type Ⅲ,n=39)according to the Wiberg classification.The Hospital for Specia Surgery knee score,Feller score,and incidence of anterior knee pain,as well as radiologic data(patellar index,patellar tilt angle,and lateral patellofemoral angle)were compared among the three groups.RESULTS AND CONCLUSION:(1)There was no significant difference in the Feller patellar score and Hospital for Specia Surgery score at the final postoperative follow-up in patients with Wiberg type Ⅰ,type Ⅱ,and type Ⅲ patellae(P＞0.05).(2)Although there was no significant difference in the incidence of anterior knee pain among the three groups at the final follow-up,patients with type Ⅲ patellae were significantly more likely to have experienced anterior knee pain early in the postoperative period.(3)Different patellar morphologies can improve patellar position to some extent after unicompartmental knee arthroplasty,but type Ⅲ had greater patellar tilt than types Ⅰ and Ⅱ,both preoperatively and postoperatively.(4)This finding highlights the need for tailored morphological adjustments to the Wiberg Ⅲ patella during unicompartmental knee arthroplasty to improve surgical outcomes.

BACKGROUND:Unicompartmental knee arthroplasty effectively addresses medial knee osteoarthritis.The accurate tibial component rotational alignment is crucial to ensure the best surgical outcome.OBJECTIVE:To assess the impact of tibial component rotational alignment on short-term outcomes after unicompartmental knee arthroplasty in medial knee osteoarthritis patients.METHODS:From February 2021 to February 2023,91 patients who underwent mobile-bearing unicompartmental knee arthroplasty in Affiliated Hospital of Xuzhou Medical University were selected.According to the rotational alignment of tibial prosthesis relative to Akagi's line measured by postoperative CT,the patients were divided into three groups.Group A had internal rotation greater than 0°,group B had external rotation from 0° to 5°,and group C had external rotation greater than 5°.The knee range of motion,Oxford knee score,and forgotten joint score were recorded and compared among three groups preoperatively,at the last follow-up after surgery.RESULTS AND CONCLUSION:(1)At the last follow-up after surgery,the Oxford knee score of group B was significantly lower than that of group A(P=0.003)and group C(P=0.025).The knee range of motion of group B was higher than that of group A(P=0.011)and group C(P=0.024),and the forgotten joint score of group B was higher than that of group A(P=0.001)and group C(P=0.049).(2)The forgotten joint score in group C was significantly higher than that in group A at the last follow-up(P=0.044).(3)Patients were followed up for 12-36 months after arthroplasty.There were 2 and 7 cases of postoperative knee pain in groups B and C,and the difference was statistically significant(P=0.033).(4)It is indicated that external rotation of the tibial prosthesis by 0°-5° is ideal for achieving satisfactory short-term clinical outcomes,so we should avoid intraoperative tibial prosthesis malrotation.

Objective To investigate the effect of PIAS3 on glucose fluctuation-induced oxidative stress and mito-chondrial dysfunction in rat cardiomyocytes.Methods H9c2 were cultured in vitro,and divided into normal glucose control group(Control),mannitol-induced osmotic pressure control group(MG),constant high glucose group(HG),intermittent hyperglycemia group(IHG),IHG+siRNA NC group,and IHG+PIAS3 siRNA group.Cell proliferation was assessed using CCK-8 assay.LDH release,MDA and GSH levels,as well as SOD activity,were detected using corresponding kits.Mitochondrial membrane potential was evaluated via JC-1 staining combined with flow cytometry.ROS levels in cells and mitochondria were determined using DCFH-DA and MitoSOX staining,re-spectively.Protein expression of PI3K,p-PI3K,AKT,and p-AKT was analyzed by Western blot.Results Com-pared with the control group,intermittent hyperglycemia promoted oxidative stress and mitochondrial dysfunction,significantly upregulated PIAS3 expression(P＜0.001)and downregulated p-PI3K and p-AKT protein levels(P＜0.001).Knockdown of PIAS3 significantly alleviated oxidative stress and mitochondrial dysfunction induced by glucose fluctuations,and increased p-PI3K and p-AKT protein levels(P＜0.001).Conclusions Knockdown of PIAS3 may alleviate glucose fluctuation-induced oxidative stress and mitochondrial dysfunction in ratcardiomyocytes by activating the PI3K/AKT signaling pathway.

Objective:To investigate the immuno-inflammatory regulatory effects of the Qing Gan San Jie Xiao Ying Formula(QGSJXYF)on Hashimoto's thyroiditis(HT)by modulating thyroid cell-derived exosomes to provide experi-mental evidence for its immunomodulatory mechanisms.Methods:Nthy-ori-3-1 thyroid cells were treated with QGSJXYF-medicated serum,with untreated cells serving as controls.Exosomes from both groups were extracted and analyzed using nanoparticle tracking analysis(NTA),transmission electron microscopy(TEM),and Western blot to assess concentration,size distribution,morphology,and the expression of characteristic exosomal markers.An inflammatory model of human T lymphocytes(H9)was established and co-incubated with normal exosomes(EXO-C group)or QGSJXYF-treated exosomes(EXO-T group).The levels of inflammatory cytokines in H9 cells were measured using Western blot(WB)and ELISA.Results:Exosome characterization showed that the particle concentration of Nthy-ori-3-1 cell-derived exosomes in both the control and QGSJXYF groups ranged from 1×109 to 1×1011/mL,with particle diameters between 80～300 nm.The exosomes exhibited a typical spherical or cup-shaped morphology with positive expression of TSG101,CD63,and HSP70.Compared with the inflammation model group and the EXO-C group,the EXO-T group significantly reduced the intracellular expression of IL-17A protein in H9 cells(P<0.05)and suppressed IL-17 and IL-6 levels in the cell supernatant(P<0.01).Conclusion:QGSJXYF may exert its anti-inflammatory and thyroid-protective effects by modulating the functional state of thyroid cell-derived exosomes,regulating the inflamma-tory microenvironment,and inhibiting the expression of inflammatory cytokines associated with Hashimoto's thyroiditis.

Colorectal cancer(CRC)is a prevalent malignancy of the digestive tract in China;it exhibits aggressive progression that is closely associated with tumor microenvironment(TME)modulation.Tumor-associated macrophages,which are pivotal immunomodulatory com-ponents of the TME,demonstrate remarkable functional heterogeneity.Among these,secreted phosphoprotein 1-positive tumor-associated macrophages(SPP1+TAMs)represent a distinct subset with well-characterized protumorigenic properties.Evidence indicates that SPP1+TAMs exhibit a unique spatial distribution pattern in CRC tissues,with pronounced enrichment at the invasive tumor front and metastatic niches.Through the secretion of SPP1 and other effector molecules,this subset orchestrates multifaceted oncogenic processes,including tumor cell adhesion,migration,angiogenesis,and metastatic dissemination.This review systematically elucidates the spatial distribution,molecular reg-ulatory mechanisms,and clinical implications of SPP1+TAMs in CRC,thereby providing a theoretical foundation for developingnovel diagnost-ic biomarkers and targeted therapeutic strategies.