Objective To explore the value of combining quantitative indicators of rectus femoris muscle obtained through ultrasound technology with clinical data in identifying protein energy wasting(PEW)in patients with chronic kidney disease(CKD)undergoing maintenance hemodialysis.Methods 90 CKD patients undergoing regular hemodialysis treatment in Guangzhou Red Cross Hospital from November 2023 to August 2024 were selected and divided into experimental group(30 cases with PEW)and control group(60 cases without PEW)according to PEW diagnostic criteria.Clinical indicators and quadriceps ultrasound quantitative data were measured and compared between two groups.Independent predictors of PEW were screened through multivariate Logistic regression analysis.A prediction model was constructed and receiver operating characteristic curve were drawn to evaluate its diagnostic performance.Results Significant differences were observed between experimental group and control group in body mass index(BMI),serum albumin,age,gender,rectus femoris thickness,and cross-sectional area(P＜0.05).Multivariate Logistic regression analysis showed that BMI and rectus femoris thickness were independent factors affecting PEW in CKD patients.A diagnostic model based on these two indicators,Logit(P)=1/[1+e-(10.292-0.378×BMI-2.992×rectus femoris thickness)],demonstrated good diagnostic performance(area under the curve=0.853,95％CI:0.764-0.943,P＜0.05).Conclusion Ultrasound assessment of rectus femoris thickness can objectively reflect the degree of skeletal muscle atrophy in CKD patients,and the prediction model constructed in combination with BMI helps in early identification of PEW and clinical intervention decision-making.

Objective To investigate the effects of acteoside in a rat model on of diabetic nephropathy(DN)and high-glucose-induced glomerular mesangial cells(GMCs),focusing on the role of acteoside in the silent information regulator 1(SIRT1)/nuclear factor eryth-roid 2-related factor 2(Nrf2)signaling pathway.Methods GMCs were cultured under normal glucose conditions or stimulated with high glucose.Fibrosis,oxidative stress,SIRT1 and Nrf2 protein expression,and mitochondrial structure were assessed in four groups:normal glucose,high glucose,high glucose+acteoside,and high glucose+acteoside+SIRT1 inhibitor(hsa62).DN was induced in rats via intraperitoneal streptozotocin injection,and animals were divided into control,model,and acteoside-treated groups.Pathological kidney changes,blood glucose changes,renal function indices,and protein expression of SIRT1 and Nrf2 were evaluated.Results Compared with controls,DN model rats showed significantly elevated fasting blood glucose,serum creatinine,blood urea nitrogen,and 24-h urinary protein levels of rats in the model group were significantly increased(P＜0.01).GMCs exhibited increased fibrosis,oxidative stress,and mitochondrial damage.Acteoside treatment significantly improved all measured parameters(P＜0.01);and mitigated mitochondrial injury.In vitro,acteoside reduced high-glucose-induced fibrosis and oxidative stress in GMCs,effects that were reversed by SIRT1 inhibition.Western blotting confirmed upregulation of SIRT1 and Nrf2 expression in both treated rat kidney tissues and GMCs(P＜0.01).Conclusion Acteoside alleviates glomerular fibrosis and oxidative stress in DN by activating the SIRT1/Nrf2 signaling pathway,suggesting its potential therapeutic agent for DN.

Objective To explore the value of combining quantitative indicators of rectus femoris muscle obtained through ultrasound technology with clinical data in identifying protein energy wasting(PEW)in patients with chronic kidney disease(CKD)undergoing maintenance hemodialysis.Methods 90 CKD patients undergoing regular hemodialysis treatment in Guangzhou Red Cross Hospital from November 2023 to August 2024 were selected and divided into experimental group(30 cases with PEW)and control group(60 cases without PEW)according to PEW diagnostic criteria.Clinical indicators and quadriceps ultrasound quantitative data were measured and compared between two groups.Independent predictors of PEW were screened through multivariate Logistic regression analysis.A prediction model was constructed and receiver operating characteristic curve were drawn to evaluate its diagnostic performance.Results Significant differences were observed between experimental group and control group in body mass index(BMI),serum albumin,age,gender,rectus femoris thickness,and cross-sectional area(P＜0.05).Multivariate Logistic regression analysis showed that BMI and rectus femoris thickness were independent factors affecting PEW in CKD patients.A diagnostic model based on these two indicators,Logit(P)=1/[1+e-(10.292-0.378×BMI-2.992×rectus femoris thickness)],demonstrated good diagnostic performance(area under the curve=0.853,95％CI:0.764-0.943,P＜0.05).Conclusion Ultrasound assessment of rectus femoris thickness can objectively reflect the degree of skeletal muscle atrophy in CKD patients,and the prediction model constructed in combination with BMI helps in early identification of PEW and clinical intervention decision-making.

Objective To investigate the effects of acteoside in a rat model on of diabetic nephropathy(DN)and high-glucose-induced glomerular mesangial cells(GMCs),focusing on the role of acteoside in the silent information regulator 1(SIRT1)/nuclear factor eryth-roid 2-related factor 2(Nrf2)signaling pathway.Methods GMCs were cultured under normal glucose conditions or stimulated with high glucose.Fibrosis,oxidative stress,SIRT1 and Nrf2 protein expression,and mitochondrial structure were assessed in four groups:normal glucose,high glucose,high glucose+acteoside,and high glucose+acteoside+SIRT1 inhibitor(hsa62).DN was induced in rats via intraperitoneal streptozotocin injection,and animals were divided into control,model,and acteoside-treated groups.Pathological kidney changes,blood glucose changes,renal function indices,and protein expression of SIRT1 and Nrf2 were evaluated.Results Compared with controls,DN model rats showed significantly elevated fasting blood glucose,serum creatinine,blood urea nitrogen,and 24-h urinary protein levels of rats in the model group were significantly increased(P＜0.01).GMCs exhibited increased fibrosis,oxidative stress,and mitochondrial damage.Acteoside treatment significantly improved all measured parameters(P＜0.01);and mitigated mitochondrial injury.In vitro,acteoside reduced high-glucose-induced fibrosis and oxidative stress in GMCs,effects that were reversed by SIRT1 inhibition.Western blotting confirmed upregulation of SIRT1 and Nrf2 expression in both treated rat kidney tissues and GMCs(P＜0.01).Conclusion Acteoside alleviates glomerular fibrosis and oxidative stress in DN by activating the SIRT1/Nrf2 signaling pathway,suggesting its potential therapeutic agent for DN.

OBJECTIVE To confirm the therapeutic effect of Jintiange capsules on osteoarthritis(OA) and the potential mechanism of synovial mesenchymal stem cell exosomes(SMSC-Exos) and articular chondrocytes(ACs) in the treatment of OA based on high-throughput sequencing technology. METHODS Type Ⅱ collagenase-induced OA rats were used for efficacy verification through general behavioral observation, bipedal balance difference experiment, mechanical foot reflex threshold, Micro-CT observation, and Safranin O-Fast Green staining. SMSCs and ACs were cultured in suitable concentration of drug-containing serum, and mRNA sequencing was performed on ACs in the control, model, and Jintiange capsules groups, as well as miRNA sequencing on SMSC-Exos. Differential expressed mRNAs and miRNAs were screened and target genes were predicted. The common differential expressed genes between SMSC and ACs were obtained by intersecting the differential expressed genes, and a miRNA-mRNA regulatory network was constructed using Cytoscape software. The expression trend analysis of common differential expressed genes was conducted, as well as the correlation analysis between differential expressed gene mRNA and miRNA, Micro-CT efficacy indicators, and differential expressed gene mRNA. RESULTS Under the pathological state of OA, the expression of miRNA-23a-3p, miRNA-342-3p, miRNA-146b-5p, miRNA-501-3p, and miRNA-214-3p were down-regulated, while miRNA-222-3p, miRNA-30e-3p, miRNA-676-3p, and miRNA-192-5p were up-regulated (P<0.05). The expressions of these miRNAs were significantly reversed after intervention with drug-containing serum of Jintiange capsules. There was a certain correlation between Micro-CT efficacy indicators, mRNA and miRNA. CONCLUSION Jintiange capsule has obvious efficacy in the treatment of OA, and its mechanism may be related to the promotion of SMSC-Exos targeting ACs to transport miRNA and then regulate Serpinb10, Ntn1, Il1b, Tgm2, Megf10, Il11, Cd40, Slc15a3, Pou2f2 and other genes.

Objective The aim of this study was to explore the correlation between the methylation status of microRNA let-7a-3 in esophageal squamous cell carcinoma(ESCC)and the expression of insulin-like growth factor 2(IGF-Ⅱ).Methods The methylation specific PCR(qMSP)was used to detect the methylation status of let-7a-3 in 83 cases of esophageal cancer and corre-sponding adjacent normal tissues.The enzyme linked immunosorbent assay(ELISA)was used to detect the expression of IGF-Ⅱ in plasma.Results The degree of let-7a-3 methylation in cancer tissues of 83 patients with ESCC was significantly higher than that in normal tissues adjacent to cancer(P<0.001).The expression of IGF-Ⅱ in the plasma of 83 patients with ESCC was positively corre-lated with the methylation degree of let-7a-3,which was statistically significant(r=0.600,P<0.001).Conclusion microRNAlet-7a-3 may participate in the occurrence and progression of ESCC by regulating the methylation of downstream molecules,which is of great significance for understanding the mechanisms of ESCC development and providing a basis for the diagnosis and prognosis of ESCC.

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.

Objective To assess the safety of femoral artery puncture procedures guided by X-ray and ultrasound among elderly patients.Methods A total of 480 patients undergoing transcatheter interventional treatment for cardiovascular and cerebrovascular diseases through the femoral ar-tery in our hospital between January 2016 and December 2022 were enrolled in the study.Of them,326 patients receiving femoral artery puncture guided by X-ray fluoroscopy were assigned into X-ray group,while the other 154 patients guided by vascular Doppler ultrasound were into ultrasound group.With propensity score matching(PSM)in a ratio of 1∶1,finally 270 patients were included.Their general clinical data,success rate of puncture,puncture site,and incidence of vascular complications were compared between the two groups.Multivariate logistic regression analysis was used to identify the risk factors for vascular complications.Results Before PSM,there were no statistical differences in the mean distance from the skin fold to the bifurcation of the common femoral artery(2.5±1.0 cm vs 2.4±0.8 cm)or the distance from the fold to the in-guinal ligament(6.4±1.4 cm vs 6.3±1.7 cm)between the X-ray group and the ultrasound group(P＞0.05).After PSM,the X-ray group exhibited an obviously higher incidence of puncture points below the common femoral artery than the ultrasound group(14.8％vs 6.7％,P＜0.05),but no significant differences were observed in the one-time success rate of puncture or the occur-rence of vascular complications between the two groups(P＞0.05).Multivariate logistic regres-sion analysis indicated that the presences of non-common femoral artery and femoral artery calci-fication at the puncture site was independent risk factors for vascular complications(OR=8.379,95％CI:3.561-19.717;OR=3.922,95％CI:1.664-9.242).Conclusion There is no statistical disparity in safety between X-ray-versus ultrasound-guided femoral artery puncture procedures.Cli-nicians should choose appropriate puncture procedure or combine them together based on individual con-dition of patients.

Porcine deltacoronavirus(PDCoV)is one of the important pathogens associated with swine viral diarrhea,which results in enormous economic losses in the pig industry.Among the known encoding proteins,the nucleocapsid(N)protein is relatively conserved and serves as the dominant antigen of coronaviruses,making it an ideal candidate for the early and accurate diagnosis of infection.In this study,the PDCoV N protein was induced and expressed in a prokaryotic system using IPTG treatment at low temperatures.Six-week-old BALB/c mice were immunized with the recombinant PDCoV N protein purified by nickel column.Isolated spleen cells were harvested and fused with SP2/0 cells once the serum titers reached 1∶10 000,as determined by indirect ELISA.The positive monoclonal hybridoma with the highest titer of secretory antibodies were screened af-ter three rounds of limited dilution.Ultimately,indirect immunofluorescent assay(IFA)and West-ern blot were employed to confirm the characterization of the antibodies,and the subtypes of the heavy and light chains of the antibodies were determined.One strain of hybridoma against the PD-CoV N protein,designated as 2G12,was obtained,with a titer approaching 1∶40 960.The IFA as-say showed PDCoV was specifically bound to 2G12.Furthermore,the Western blot assay further showed PDCoV in the supernatant of infected cells or recombinant PDCoV N protein,reacted well with 2G12.The subtype of MAbs was determined as IgG1 with the light chains being κ.The MAbs generated against the PDCoV N protein in this study provide valuable support for the development of a diagnostic kit for PDCoV and play an important role in the functional research of the PDCoV N protein.

OBJECTIVE To optimize the heart-nourishing granule extraction process by network pharmacology and Box-Behnken response surface method. METHODS The active ingredients of heart-nourishing granule were excavated through network pharmacology and their mechanism of action in the treatment of coronary heart disease was preliminarily explored. Molecular docking technology was used to predict the binding ability of the active ingredients to the main targets. At the same time, based on the compatibility relationship between the monarch, minister, assistant and guide of the prescription, the pharmacological effects of the ingredients, and the content determination index components of each medicinal flavor in the 2020 edition of the Chinese Pharmacopoeia, the process evaluation index components of heart-nourishing granule were further determined. In addition, combined with the analytic hierarchy process to determine the weight of each component, the Box-Behnken response surface method was used to optimize the extraction process of heart-nourishing granule. RESULTS The main active components of heart-nourishing granule screened were catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin. By acting on core targets such as AKT1, IL-6, IL-1b, VEGFA, JUN and MAPK3, they regulated lipid and atherosclerosis, MAPK signaling and other pathways played a role in the treatment of coronary heart disease. Molecular docking results showed that the binding energies of active components and core targets predicted by network pharmacology were all < -5.0 kJ·mol-1. Five components catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin were calculated by analytic hierarchy process method. The weight coefficients of arginyl glycosides were 0.329 7, 0.329 7, 0.164 8, 0.109 9, and 0.065 9, respectively. The Box-Behnken response surface method obtained the optimal water extraction process: 10 times the amount of water was used to extract twice, and each time was decocted for 1.5 h. The verification test showed that the contents of the five components were consistent with the predicted values, and the RSD values were all <5%. CONCLUSION The extraction process of heart-nourishing granule based on network pharmacology and Box-Behnken response surface methodology is stable and feasible, which provided an experimental basis for its further quality improvement.