Splicing factor YBX1 regulates bone marrow stromal cell fate during aging.

Senescence and altered differentiation potential of bone marrow stromal cells (BMSCs) lead to age-related bone loss. As an important posttranscriptional regulatory pathway, alternative splicing (AS) regulates the diversity of gene expression and has been linked to induction of cellular senescence. However, the role of splicing factors in BMSCs during aging remains poorly defined. Herein, we found that the expression of the splicing factor Y-box binding protein 1 (YBX1) in BMSCs decreased with aging in mice and humans. YBX1 deficiency resulted in mis-splicing in genes linked to BMSC osteogenic differentiation and senescence, such as Fn1, Nrp2, Sirt2, Sp7, and Spp1, thus contributing to BMSC senescence and differentiation shift during aging. Deletion of Ybx1 in BMSCs accelerated bone loss in mice, while its overexpression stimulated bone formation. Finally, we identified a small compound, sciadopitysin, which attenuated the degradation of YBX1 and bone loss in old mice. Our study demonstrated that YBX1 governs cell fate of BMSCs via fine control of RNA splicing and provides a potential therapeutic target for age-related osteoporosis.

4EBP2-regulated protein translation has a critical role in high-fat diet-induced insulin resistance in hepatocytes.

A high-fat diet (HFD) plays a critical role in hepatocyte insulin resistance. Numerous models and factors have been proposed to elucidate the mechanism of palmitic acid (PA)-induced insulin resistance. However, proteomic studies of insulin resistance by HFD stimulation are usually performed under insulin conditions, leading to an unclear understanding of how a HFD alone affects hepatocytes. Here, we mapped the phosphorylation rewiring events in PA-stimulated HepG2 cells and found PA decreased the phosphorylation level of the eukaryotic translation initiation factor 4E-binding protein 2 (4EBP2) at S65/T70. Further experiments identified 4EBP2 as a key node of insulin resistance in either HFD mice or PA-treated cells. Reduced 4EBP2 levels increased glucose uptake and insulin sensitivity, whereas the 4EBP2_S65A/T70A mutation exacerbated PA-induced insulin resistance. Additionally, the nascent proteome revealed many glycolysis-related proteins translationally regulated by 4EBP2 such as hexokinase-2, pyruvate kinase PKM, TBC1 domain family member 4, and glucose-6-phosphate 1-dehydrogenase. In summary, we report the critical role of 4EBP2 in regulating HFD-stimulated insulin resistance in hepatocytes.

Artificial intelligence-assisted system for the assessment of Forrest classification of peptic ulcer bleeding: a multicenter diagnostic study.

BACKGROUND: Inaccurate Forrest classification may significantly affect clinical outcomes, especially in high risk patients. Therefore, this study aimed to develop a real-time deep convolutional neural network (DCNN) system to assess the Forrest classification of peptic ulcer bleeding (PUB). METHODS: A training dataset (3868 endoscopic images) and an internal validation dataset (834 images) were retrospectively collected from the 900th Hospital, Fuzhou, China. In addition, 521 images collected from four other hospitals were used for external validation. Finally, 46 endoscopic videos were prospectively collected to assess the real-time diagnostic performance of the DCNN system, whose diagnostic performance was also prospectively compared with that of three senior and three junior endoscopists. RESULTS: The DCNN system had a satisfactory diagnostic performance in the assessment of Forrest classification, with an accuracy of 91.2% (95%CI 89.5%-92.6%) and a macro-average area under the receiver operating characteristic curve of 0.80 in the validation dataset. Moreover, the DCNN system could judge suspicious regions automatically using Forrest classification in real-time videos, with an accuracy of 92.0% (95%CI 80.8%-97.8%). The DCNN system showed more accurate and stable diagnostic performance than endoscopists in the prospective clinical comparison test. This system helped to slightly improve the diagnostic performance of senior endoscopists and considerably enhance that of junior endoscopists. CONCLUSION: The DCNN system for the assessment of the Forrest classification of PUB showed satisfactory diagnostic performance, which was slightly superior to that of senior endoscopists. It could therefore effectively assist junior endoscopists in making such diagnoses during gastroscopy.

Renal tubular epithelial cell necroptosis promotes tubulointerstitial fibrosis in patients with chronic kidney disease.

Renal fibrosis, a common pathological manifestation of virtually all types of chronic kidney disease (CKD), ultimately predisposes patients to end-stage renal disease. However, there is no effective therapy for renal fibrosis. Our earlier studies proved that RIP3-mediated necroptosis might be an important mode of renal tubular cell death in rats with chronic renal injury. Under transmission electron microscopy (TEM), we found morphological changes in the necrosis of human renal tissue, and the percentage of necrotic cells increased significantly in patients with stages 2 and 3a CKD. Immunofluorescence analyses showed that the percentages of TUNEL+ /RIP3+ double-positive and TUNEL+ /MLKL+ double-positive tubular epithelial cells in renal tubules of patients with stages 2 and 3a CKD were significantly increased compared to those in control patients without renal disease. Immunohistochemistry analyses of renal biopsy specimens from patients with CKD revealed RIP3, MLKL, and p-MLKL upregulation in patients with stages 2 and 3a CKD, suggesting that necroptosis of renal tubular epithelial cells in CKD patients occurs, and the peak of necroptosis was in stages 2 and 3a CKD. We showed that profibrotic factor proteins (TGF-ß1, Smad2 and Smad3) and fibroblast activation markers (α-SMA and Vimentin) were specifically upregulated in stage 2 and 3a CKD patients. In addition, Pearson correlation analysis showed that the percentage of necroptotic renal tubular epithelial cells was positively correlated with TGF-ß1 and collagen-I. We also showed that RIP1/3 or MLKL inhibitors decreased the expression of RIP3, MLKL, TGF-ß1, and Smad3 in HK-2 cells treated with TNF-α. FGF-2, α-SMA, Vimentin and FN were overexpressed in the hRIFs cultured with the supernatant of necroptotic HK-2 cells, whereas necroptosis blockers (Nec-1s, GSK'872 and NSA) and TGF-ß1/Smad3 pathway antagonists (LY364947 and SIS3) reduced FGF-2, α-SMA, Vimentin and FN levels. Collectively, necroptosis of renal tubular epithelial cells in CKD patients occurs, and the peak of necroptosis was in stages 2 and 3a CKD. Renal tubular epithelial cell necroptosis mediates renal tubulointerstitial fibrosis in patients with chronic kidney disease, which is related to the TGF-ß1/Smad3 signaling pathway.

Self-Assembly of a Series of Carbazole-Based Vinyl-benzoxazole Derivatives in Gel Phases.

A series of carbazole-based vinyl-benzoxazole derivatives have been synthesized in order to verify whether X-ray diffraction (XRD) simulation can give more information about intermolecular stacking in the gel phase. It was found that their gelation capabilities were strongly dependent on the length of the alkyl chain. The compounds with shorter alkyl chains have lower critical gelation concentrations (CGCs) in nonpolar alkane and alcohols with longer carbon chains. On the other hand, compounds with long alkyl chains presented small CGCs in polar methanol. Powder XRD structure solution gave more information about intermolecular stacking than the traditional way of analyzing diffraction peaks to derive approximate molecular stacking patterns. The results verified that gelators had a similar head-to-tail π-stacking between aromatic groups in gel phases although different slipping angles existed. Moreover, ordered stacking between the alkyl chains was also present.

Chronic ß-adrenergic stress contributes to cardiomyopathy in rodents with collagen-induced arthritis.

Patients with rheumatoid arthritis (RA) have a much higher incidence of cardiac dysfunction, which contributes to the high mortality rate of RA despite anti-arthritic drug therapy. In this study, we investigated dynamic changes in cardiac function in classic animal models of RA and examined the potential effectors of RA-induced heart failure (HF). Collagen-induced arthritis (CIA) models were established in rats and mice. The cardiac function of CIA animals was dynamically monitored using echocardiography and haemodynamics. We showed that cardiac diastolic and systolic dysfunction occurred in CIA animals and persisted after joint inflammation and that serum proinflammatory cytokine (IL-1ß, TNF-α) levels were decreased. We did not find evidence of atherosclerosis (AS) in arthritic animals even though cardiomyopathy was significant. We observed that an impaired cardiac ß1AR-excitation contraction coupling signal was accompanied by sustained increases in blood epinephrine levels in CIA rats. Furthermore, serum epinephrine concentrations were positively correlated with the heart failure biomarker NT-proBNP in RA patients (r2 = +0.53, P < 0.0001). In CIA mice, treatment with the nonselective ßAR blocker carvedilol (2.5 mg·kg-1·d-1, for 4 weeks) or the specific GRK2 inhibitor paroxetine (2.5 mg·kg-1·d-1, for 4 weeks) effectively rescued heart function. We conclude that chronic and persistent ß-adrenergic stress in CIA animals is a significant contributor to cardiomyopathy, which may be a potential target for protecting RA patients against HF.

The effect of lifting load on the kinematic characteristics of lumbar spinous process in vivo.

BACKGROUND: There are limited data on the in vivo natural kinematics of the lumbar spinous process. This paper intends to explore the effect of lifting load on the in vivo movement mode of the lumbar spinous process and its biomechanical changes. METHODS: Ten asymptomatic subjects between the ages of 25 and 39 underwent CT scans of the lumbar spine in the supine position, and 3D models of L3-L5 were constructed. Using a Dual Fluoroscopy Imaging System (DFIS), instantaneous orthogonal fluoroscopic images of each subject's flexion-extension, left-right bending, and left-right rotational movements were taken under different loads (0 kg, 5 kg, 10 kg). The supine CT model was matched, using computer software, to the bony contours of the images from the two orthogonal views, so that the instantaneous 3D vertebral position at each location could be quantified. A Cartesian coordinate system was ultimately constructed at the tip of the spinous process to obtain the 6DOF kinematic data of the spinous process. RESULTS: In different postural movements of the trunk, there was no significant difference in the rotation angle and translation range of the lumbar spinous process under different loads (P > 0.05). In flexion to extension motion, spinous processes mainly rotate < 4° along the medial and lateral axes and translate < 4 mm along the craniocaudal direction. In the left-right bending motion, spinous processes mainly rotate < 5° along the anterior and posterior axes, and the translation is mainly coupling < 2 mm. In the rotational motion, the spinous process is mainly coupled motion, the rotation range is less than 3°, and the translation range is less than 2 mm. The distance between spinous processes measured in the supine position was 6.66 ± 2.29 mm at L3/4 and 5.08 ± 1.57 mm at L4/5. CONCLUSION: The in vivo kinematics of the lumbar spinous process will not change significantly with increasing low load. In complex motion, the spinous process is dominated by coupling motion.

The role of autophagy in bone homeostasis.

Autophagy is an evolutionarily conserved intracellular process and is considered one of the main catabolism pathways. In the process of autophagy, cells are digested nonselectively or selectively to recover nutrients and energy, so it is regarded as an antiaging process. In addition to the essential role of autophagy in cellular homeostasis, autophagy is a stress response mechanism for cell survival. Here, we review recent literature describing the pathway of autophagy and its role in different bone cell types, including osteoblasts, osteoclasts, and osteocytes. Also discussed is the mechanism of autophagy in bone diseases associated with bone homeostasis, including osteoporosis and Paget's disease. Finally, we discuss the application of autophagy regulators in bone diseases. This review aims to introduce autophagy, summarize the understanding of its relevance in bone physiology, and discuss its role and therapeutic potential in the pathogenesis of bone diseases such as osteoporosis.

BirA*-protein A fusion protein based BioEnhancer amplifies western blot immunosignal.

Western blot (protein immunoblot) is a widely used analytical technique in molecular biology. Utilizing the specific recognizing primary antibody, proteins immobilized on various matrix are investigated by subsequent visualization steps, for example, by the horse radish peroxidase conjugated secondary antibody incubation. Methods to improve the sensitivity in protein identification or quantification are appreciated by biochemists. Herein, we report a new strategy to amplify Western blot signals by constructing a probe with proximal labeling and IgG targeting abilities. The R118G mutation attenuated the biotin-AMP binding affinity of the bacterial biotin ligase BirA*, offering a proximity-dependent labeling ability, which could be used as a signal amplifier. We built a BirA*-protein A fusion protein (BioEnhancer) that specifically binds to IgG and adds biotin tags to its proximal amine groups, enhancing the immunosignal of target proteins. In our experiments, the BioEnhancer system amplified the immunosignal by tenfold compared to the standard western blot. Additionally, our strategy could couple with other signal enhancement methods to further increase the western blot sensitivity.

TRAM couples endocytosis of Toll-like receptor 4 to the induction of interferon-beta.

Toll-like receptor 4 (TLR4) induces two distinct signaling pathways controlled by the TIRAP-MyD88 and TRAM-TRIF pairs of adaptor proteins, which elicit the production of proinflammatory cytokines and type I interferons, respectively. How TLR4 coordinates the activation of these two pathways is unknown. Here we show that TLR4 activated these two signaling pathways sequentially in a process organized around endocytosis of the TLR4 complex. We propose that TLR4 first induces TIRAP-MyD88 signaling at the plasma membrane and is then endocytosed and activates TRAM-TRIF signaling from early endosomes. Our data emphasize a unifying theme in innate immune recognition whereby all type I interferon-inducing receptors signal from an intracellular location.

The Clinical Significance of Dickkopf Wnt Signaling Pathway Inhibitor Gene Family in Head and Neck Squamous Cell Carcinoma.

BACKGROUND Dickkopf Wnt signaling pathway inhibitor (DKK) gene family, which is known to inhibit the Wnt regulation process, is widely found in cancers. However, the roles and functions of specific family members in head and neck squamous cell carcinoma (HNSCC) are still unclear. MATERIAL AND METHODS Online bioinformatics tools (Oncomine, UALCAN, Kaplan-Meier plotter, GEPIA, Metascape, and STRING) were used to analyze the relationships between distinct DKKs and HNSCC. The transcriptome expression, clinical association, functions, pathways, and protein-protein interaction networks of DKKs in HNSCC were explored. RESULTS The mRNA expression of DKK1, DKK3, and Dickkopf-like acrosomal protein 1 (DKKL1) in HNSCC was significantly higher than in normal tissues, while that of DKK4 was lower. The mRNA expression of DKK1, DKK3, and DKKL1 was elevated in higher-grade HNSCC. The mRNA expression of DKK1 and DKK3 was elevated in human papillomavirus (HPV)-negative HNSCC, while DKKL1 had a higher mRNA expression in HPV-positive HNSCC. In addition, DKK1 was significantly associated with unfavorable overall survival in HNSCC patients. DKK3 was more likely to be a negative factor for the 5-year survival rate, while DKK4 was the opposite. DKK1 function was mainly enriched in GTPase-mediated signal transduction. Porcupine O-acyltransferase, a key regulator of the Wnt signaling pathway, was also associated with DKK1 in the protein-protein interaction network. CONCLUSIONS With regard to improving the therapeutic strategies of HNSCC in the future, DKK1 could be an unfavorable prognostic biomarker. DKK3, DKK4, and DKKL1 might be potential biomarkers for HNSCC.

In-vitro differentiation of early pig spermatogenic cells to haploid germ cells.

Spermatogonial stem cells (SSCs) self-renew and contribute genetic information to the next generation. Pig is wildly used as a model animal for understanding reproduction mechanisms of human being. Inducing directional differentiation of porcine SSCs may be an important strategy in exploring the mechanisms of spermatogenesis and developing better treatment methods for male infertility. Here, we established an in-vitro culture model for porcine small seminiferous tubule segments, to induce SSCs to differentiate into single-tail haploid spermatozoa. The culture model subsequently enabled spermatozoa to express the sperm-specific protein acrosin and oocytes to develop to blastocyst stage after round spermatid injection. The addition of retinoic acid (RA) to the differentiation media promoted the efficiency of haploid differentiation. RT-PCR analysis indicated that RA stimulated the expression of Stra8 but reduced the expression of NANOS2 in spermatogonia. Genes involved in post-meiotic development, transition protein 1 (Tnp1) and protamine 1 (Prm1) were upregulated in the presence of RA. The addition of an RA receptor (RAR) inhibitor, BMS439, showed that RA enhanced the expression of cAMP responsive-element binding protein through RAR and promoted the formation of round spermatids. We established an efficient culture system for in-vitro differentiation of pig SSCs. Our study represents a model for human testis disease and toxicology screening. Molecular regulators of SSC differentiation revealed in this study might provide a therapeutic strategy for male infertility.

Iodine-catalyzed guanylation of amines with N,N'-di-Boc-thiourea.

Herein, we report that iodine-catalyzed guanylation of primary amines can be accomplished with N,N'-di-Boc-thiourea and TBHP to afford the corresponding guanidines in 40-99% yields. Oxidation of the HI byproduct by TBHP eliminates the need for an extra base to prevent the protonation of substrates and makes the reaction especially useful for both electronically and sterically deactivated primary anilines.

The effect of phospho-peptide on the stability of gold nanoparticles and drug delivery.

BACKGROUND: Gold nanoparticles (AuNPs) have been proposed for many applications in medicine and bioanalysis. For use in all these applications, maintaining the stability of AuNPs in solution by suppressing aggregation is paramount. Herein, the effects of amino acids were investigated in stabilizing AuNPs by rationally designed peptide scaffolds. RESULTS: Compared to other tested amino acids, phosphotyrosine (pY) significantly stabilized AuNPs. Our results indicated that pY modified AuNPs presented a high level of stability in various solutions, and had good biocompatibility. When a pY-peptide was used in stabilizing AuNPs, the phosphate group could be removed by phosphatases, which subsequently caused the aggregation and the cargo release of AuNPs. In vitro study showed that AuNPs formed aggregation in a phosphatase concentration depending manner. The aggregation of AuNPs was well correlated with the enzymatic activity (R2 = 0.994). In many types of cancer, a significant increase in phosphatases has been observed. Herein, we demonstrated that cancer cells treated with pY modified AuNPs in conjunction with doxorubicin killed SGC-7901 cells with high efficiency, indicating that the pY peptide stabilized AuNPs could be used as carriers for targeted drug delivery. CONCLUSION: In summary, pY peptides can act to stabilize AuNPs in various solutions. In addition, the aggregation of pY-AuNPs could be tuned by phosphatase. These results provide a basis for pY-AuNPs acting as potential drug carriers and anticancer efficacy.

Early Response of Hepatocellular Carcinoma to Chemoembolization: Volume Computed Tomography Liver Perfusion Imaging as a Short-Term Response Predictor.

OBJECTIVE: The aim of this study was to investigate the efficacy of lesion-oriented whole-liver computed tomography (CT) perfusion for predicting transarterial chemoembolization early treatment response in patients with hepatocellular carcinoma (HCC). METHODS: Volume helical shuttle CT perfusion imaging on the whole liver was prospectively performed on 39 patients with 46 independent HCC lesions before target-selected chemoembolization. The therapeutic effects were assessed: responder group included lesions with a complete and partial response, whereas the nonresponder group contained stable and progressive disease. The perfusion parameter value comparison between 2 groups and receiver operating characteristic analyses were performed. RESULTS: The responders demonstrated higher hepatic arterial perfusion (HAP) and hepatic arterial perfusion index (HAPI) and lower hepatic portal perfusion (HPP) compared with the nonresponders among the 34 lesions without Vp3 or Vp4 portal vein thrombosis. In addition, HAP and HAPI had good prognostic values. CONCLUSIONS: Whole-liver CT perfusion allows for hemodynamic evaluation of HCC lesions. The HAP, HAPI, and hepatic portal perfusion values may be useful predictors of short-term therapeutic response after transarterial chemoembolization.

CT-guided localization of small pulmonary nodules using adjacent microcoil implantation prior to video-assisted thoracoscopic surgical resection.

OBJECTIVES: To describe and assess the localization of small peripheral pulmonary nodules prior to video-assisted thoracoscopic surgical (VATS) resection using the implantation of microcoils. METHODS: Ninety-two patients with 101 pulmonary nodules underwent computed tomography (CT)-guided implantation of microcoils proximal to each nodule. Patients were randomly assigned to undergo entire microcoil or leaving-microcoil-end implantations. The complications and efficacy of the two implantation methods were evaluated. VATS resection of lung tissue containing each pulmonary lesion and microcoil were performed in the direction of the microcoil marker. Histopathological analysis was performed for the resected pulmonary lesions. RESULTS: CT-guided microcoil implantation was successful in 99/101 cases, and the placement of microcoils within 1 cm of the nodules was not disruptive. There was no difference in the complications and efficacy associated with the entire implantation method (performed for 51/99 nodules) versus the leaving-microcoil-end implantation method (performed for 48/99 nodules). All nodules were successfully removed using VATS resection. Asymptomatic pneumothorax occurred in 16 patients and mild pulmonary haemorrhage occurred in nine patients. However, none of these patients required further surgical treatment. CONCLUSIONS: Preoperative localization of small pulmonary nodules using a refined percutaneous microcoil implantation method was found to be safe and useful prior to VATS resection. KEY POINTS: • An increasing number of small, indeterminate pulmonary lesions need to be characterized. • Entire microcoil and leaving-microcoil-end implantation methods were described for nodule localization. • Adjacent microcoil localization prior to video-assisted thoracoscopic surgical resection involved minimal intervention. • Preoperative microcoil localization facilitates the definitive resection of small pulmonary nodules.

Toxic assessment of the leachates of paddy soils and river sediments from e-waste dismantling sites to microalga, Pseudokirchneriella subcapitata.

The potential adverse effects of e-waste recycling activity on environment are getting increasing concern. In this work, a model alga, Pseudokirchneriella subcapitata, was employed to assess the toxic effects of the leachates of paddy soils and river sediments collected from e-waste dismantling sites. Chemical analysis of the paddy soils and river sediments and their leachates were carried out and the growth rate, chlorophyll a fluorescence and anti-oxidative systems of the alga were measured. Results showed that two leachates decreased the amount of PSII active reaction centers and affected photosynthesis performance, interfered with chlorophyll synthesis and inhibited algal growth. Some chemical pollutants in the sediments and soils such as polybrominated diphenyl ethers (PBDEs) and metals derived from e-waste recycling activity may impose oxidative stress on algae and affect the activity of anti-oxidative enzymes such as GST, SOD, CAT and APX. The leachates of both river sediments and paddy soils are potentially toxic to the primary producers, P. subcapitata and the leachate from sediments was more deleterious than that from soils.

[Advances on chemical constituents and bioactivities of genus Stellera].

Advance on chemical constituents and pharmacological activities of Stellera plants have been conducted. The chemical constituents include terpenes, coumarins, flavonoids, lignans, volatile oils, and other compounds. Pharmacological studies showed that diterpenoids and biflavones showed strong activities, such as antitumor, anti-HIV, and immune regulations. This review hopes to provide a scientific basis for further research and explorations of the medicinal values of the genus.

Excessive mechanical loading promotes osteoarthritis development by upregulating Rcn2.

Exposure of articular cartilage to excessive mechanical loading is closely related to the pathogenesis of osteoarthritis (OA). However, the exact molecular mechanism by which excessive mechanical loading drives OA remains unclear. In vitro, primary chondrocytes were exposed to cyclic tensile strain at 0.5 Hz and 10 % elongation for 30 min to simulate excessive mechanical loading in OA. In vivo experiments involved mice undergoing anterior cruciate ligament transection (ACLT) to model OA, followed by interventions on Rcn2 expression through adeno-associated virus (AAV) injection and tamoxifen-induced gene deletion. 10 µL AAV2/5 containing AAV-Rcn2 or AAV-shRcn2 was administered to the mice by articular injection at 1 week post ACLT surgery, and Col2a1-creERT: Rcn2flox/flox mice were injected with tamoxifen intraperitoneally to obtain Rcn2-conditional knockout mice. Finally, we explored the mechanism of Rcn2 affecting OA. Here, we identified reticulocalbin-2 (Rcn2) as a mechanosensitive factor in chondrocytes, which was significantly elevated in chondrocytes under mechanical overloading. PIEZO type mechanosensitive ion channel component 1 (Piezo1) is a critical mechanosensitive ion channel, which mediates the effect of mechanical loading on chondrocytes, and we found that increased Rcn2 could be suppressed through knocking down Piezo1 under excessive mechanical loading. Furthermore, chondrocyte-specific deletion of Rcn2 in adult mice alleviated OA progression in the mice receiving the surgery of ACLT. On the contrary, articular injection of Rcn2-expressing adeno-associated virus (AAV) accelerated the progression of ACLT-induced OA in mice. Mechanistically, Rcn2 accelerated the progression of OA through promoting the phosphorylation and nuclear translocation of signal transducer and activator of transcription 3 (Stat3).

Application of buried auricular point combined with Wenjing Sanhan prescription in arteriosclerosis obliterans patients with resting pain.

BACKGROUND: Research on the combined use of ear acupoint embedding beans and warming meridians with cold-dispersing formulas for alleviating resting pain in patients with arteriosclerosis obliterans (ASO) remains limited. AIM: To explore the therapeutic efficacy of auricular point embedding beans combined with Wenjing Sanhan prescription in alleviating resting pain in patients with lower-limb ASO. METHODS: A total of 100 patients with ASO experiencing resting pain who were treated at our hospital from January 2022 to January 2023 were enrolled. They were randomly allocated into two groups using a double-blind approach. The control group was treated using a warming meridian with a cold-dispersing formula, while the study group received additional treatment with ear acupoint embedding beans. The clinical efficacy, ankle-brachial artery pressure ratio, hemorheological indicators, and traditional Chinese medicine symptom scores were compared between the two groups. RESULTS: The clinical efficacy rate in the study group was significantly higher (94.00%) than that in the control group (72.00%, P < 0.05). Moreover, the ankle-brachial artery pressure ratio was significantly higher in the study group after treatment (P < 0.05). Hemorheological parameters, including whole blood viscosity, plasma viscosity (1.83 ± 0.11) mPa/s, fibrinogen levels (3.30 ± 0.21) g/L, platelet adhesion rate (49.87% ± 10.51%), and erythrocyte aggregation index (1.79 ± 0) were improved in the study group compared to the control group. In addition, the scores for decreased skin temperature (1.41 ± 0.26), intermittent claudication (1.30 ± 0.20), and resting pain (1.23 ± 0.31) were significantly lower in the study group than those in the control group (all P < 0.05). The level of oxidative stress in the study group also exhibited significant improvement (P < 0.05), and the levels of inflammatory factors were considerably lower than those in the control group. CONCLUSION: The combination of ear point embedding beans and Wenjing Sanhan prescription demonstrates promising clinical efficacy in alleviating resting pain associated with ASO.