Based on the theory of evidence-based medicine （EBM）， this paper systematically constructed a multi-dimensional evaluation framework for traditional Chinese medicine （TCM） appropriate technologies， encompassing three core dimensions including evidence， practitioner， and patient. For the current practical challenges in the promotion of TCM technologies such as lack of high-quality evidence， inconsistent operational standards， and varying patient acceptance， the paper proposed the integration of published literature evidence and real-world research data to construct a scientific and applicable evaluation pathway. Regarding the evidence dimension， it emphasizes syste-matic assessment of effectiveness， safety， and economic efficiency， introducing methods like the target trial emulation framework to enhance evidence quality； for the practitioner dimension， it suggests developing multi-aspects competency evaluation tools based on educational background， training assessment， and clinical outcomes； for the patient dimension， it recommends designing patient acceptance assessment tools by considering factors such as technical characte-ristics， expected efficacy， patient-practitioner interaction， and the availability of alternative treatments. The purpose of the above measures is to provide methodological support for the standardized popularization and precision application of TCM appropriate technologies.

ObjectiveTo investigate the effect and molecular mechanism of exosomes derived from bone marrow mesenchymal stem cells（BMSC-exos） modified with Bushen Yisui capsule（BSYS）-containing serum on promoting the differentiation and maturation of OLN-93 oligodendrocytes by regulating miR-15b/Wnt signaling pathway. MethodsOLN-93 cells were divided into 5 groups， including the normal（NC） group， BMSC-exos group， BSYS-BMSC-exos group， BSYS-BMSC+LV-miR-15b-5p inhibitor-exos group， and BSYS-BMSC+LV-miR-15b-5p NC-exos group. DiR staining was used to observe the uptake of Exos by OLN-93 cells. The effective dosage of BSYS-BMSC-exos on OLN-93 cells was assessed by cell proliferation and activity assay（CCK-8）. Stable BMSCs lentiviral transfection strains were established to inhibit miR-15b-5p expression in both BMSCs and their exos， and transfection efficiency was verified by real-time fluorescent quantitative polymerase chain reaction（Real-time PCR） detection of miR-15b-5p. The expressions of 2′，3′-cyclic nucleotide 3′-phosphodiesterase（CNPase） and myelin proteolipid protein（PLP） in OLN-93 cells were detected by immunocytochemistry（ICC） and Western blot. The mRNA expressions of miR-15b-5p and Wnt3a in OLN-93 cells were detected by Real-time PCR， and the protein expression of Wnt3a was measured by Western blot. The expression levels of key molecules in the Wnt/β-catenin signaling pathway of OLN-93 cells， including glycogen synthase kinase（GSK）-3β， β-catenin， and T-cell specific transcription factor 4/transcription factor 7-like 2（TCF4/TCF7L2）， were measured by Real-time PCR and Western blot. ResultsDiR-labeled Exos were efficiently taken up by OLN-93 cells. The CCK-8 assay results indicated that 20 mg·L^-1 of BSYS-BMSC-exos exhibited the most significant effect in enhancing OLN-93 cell viability（P<0.01） and this dosage was selected for subsequent experiments. Following lentiviral transfection of BMSCs， Real-time PCR results revealed that miR-15b-5p was significantly suppressed in BMSCs（P<0.01）， and miR-15b-5p was also notably inhibited in BSYS-BMSC-exos（P<0.01）. ICC analysis further revealed an increase in the number of differentiated， mature CNPase and PLP-positive cells following BSYS-BMSC-exos treatment（P<0.01）. Western blot results demonstrated that the protein expression of CNPase and PLP was significantly enhanced with BSYS-BMSC-exos treatment（P<0.01）. Additionally， BSYS-BMSC-exos also increased the expression levels of miR-15b-5p and p-β-catenin proteins in OLN-93 cells， while decreased the mRNA and protein expressions of Wnt3a， as well as the mRNA expressions of β-catenin and TCF4/TCF7L2， and the protein expression level of p-GSK-3β（Ser9） was significantly reduced（P<0.05， P<0.01）. After the transfection of miR-15b-5p inhibitor into BSYS-BMSC-exos， the above effects were significantly diminished（P<0.05， P<0.01）. ConclusionBSYS-BMSC-exos facilitate the differentiation and maturation of OLN-93 cells， and its mechanism is related to the upregulation of miR-15b-5p in OLN-93 cells， which inhibits the expression of Wnt3a and thereby suppresses the Wnt signaling pathway.

ObjectiveTo investigate the intervention effects and mechanisms of the flavonoid hyperoside （Hyp） on lipopolysaccharide （LPS）-induced inflammation in the zebrafish model. MethodsZebrafish larvae were either microinjected with 0.5 g·L^-1 LPS or immersed in 1 g·L^-1 LPS for the modeling of inflammation. The larvae were then treated with Hyp at 25， 50， and 100 mg·L^-1 through immersion for four consecutive days. The inflammatory phenotypes were assessed by analyzing the mortality rate， malformation rate， body length， and yolk sac area ratio. Behavioral tests were conducted to evaluate the inflammatory stress responses， and macrophage migration was observed by fluorescence microscopy. Additionally， the mRNA levels of inflammation-related genes， including interleukin-1β （IL-1β）， interleukin-6 （IL-6）， chemokine C-C motif ligand 2 （CCL2）， chemokine C-X3-C motif receptor 1 （CX3CR1）， chemokine C-C motif receptor 2 （CCR2）， and genes associated with the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor-kappa B （NF-κB） signaling pathway， were measured by Real-time quantitative polymerase chain reaction（Real-time PCR）. ResultsCompared with the pure water injection group， the model group exhibited increased mortality， malformation rates and yolk sac area ratio （P0.01）， reduced body length （P0.01）， increased total swimming distance and high-speed swimming duration （P0.01）， and up-regulated mRNA levels of TLR4， MyD88， NF-κB， IL-1β， IL-6， CCL2， CX3CR1， and CCR2 （P0.01）. Hyp at low， medium and high doses， as well as aspirin， reduced the mortality and malformation rates （P0.05，P0.01）， increased the body length （P0.05，P0.01）， decreased the yolk sac area ratio （P0.01）， reduced the high-speed swimming duration （P0.01）， and down-regulated the mRNA levels of TLR4， MyD88， NF-κB， IL-1β， IL-6， CCL2， CX3CR1， and CCR2 （P0.05，P0.01） compared with the model group. ConclusionHyp may modulate the TLR4/MyD88/NF-κB pathway to ameliorate inflammatory phenotypes and alleviate stress conditions in zebrafish， thereby exerting the anti-inflammatory effect.

Porcine epidemic diarrhea virus (PEDV) is an intestinal coronavirus that can cause porcine epidemic diarrhea, leading to diarrhea, vomiting, weight loss, and even death in piglets. Due to the diversity of PEDV strains, traditional vaccines are difficult to sustainably and effectively prevent and control PEDV. This article reviews the strategies and mechanisms of active substances in regulating intracellular signaling pathways, viral proteins, and microbial metabolites to enhance the host immune function against PEDV. It emphasizes the prevention of PEDV resistance and the potential harm of PEDV breaking through interspecies barriers to the human society, aiming to provide reliable theoretical support for the development of new antiviral drugs or vaccines.
Porcine epidemic diarrhea virus/immunology* ; Animals ; Swine ; Swine Diseases/prevention & control* ; Antiviral Agents/pharmacology* ; Coronavirus Infections/virology* ; Viral Vaccines/immunology* ; Humans ; Signal Transduction

Objective To construct a logistic regression prediction model for stress ulcer(SU)after cerebral hemorrhage.Methods A total of 230 patients with cerebral hemorrhage admitted to our hospital from January 2020 to January 2023 were prospectively selected as the study subjects.They were randomly di-vided into a training group and a validation group using a random number table method,with 115 patients in each group.The incidence of postoperative SU was statistically compared between the two groups.The least absolute shrinkage and selection operator(Lasso)and logistic regression were used to analyze the influencing factors of SU after cerebral hemorrhage,and a logistic regression prediction model was established and valida-ted.Results The incidence of SU was 19.13%in the training group and 20.00%in the validation group.In-crement of age,blood loss≥30 mL,higher levels of neutrophil-to-lymphocyte ratio(NLR),heat shock protein 70(HSP70)and HSP90 were identified as independent risk factors for SU after cerebral hemorrhage(P<0.05),while lower levels of Glasgow Coma Scale(GCS)score and albumin(Alb)were protective factors(P<0.05).The prediction model was logit(P)=0.409×age+1.288×blood loss-1.335×GCS score-1.126×Alb+0.452×NLR+1.483×HSP70+1.593×HSP90-10.325.The areas under the receiver operat-ing characteristic(ROC)curve(AUC)for the training group and the validation group were 0.845(95%CI:0.765-0.906)and 0.855(95%CI:0.777-0.913),respectively.The sensitivities were 81.82%and 90.91%,and the specificities were 76.34%and 70.97%,respectively.Conclusion A logistic regression prediction model was successfully constructed,which has certain predictive value for SU after cerebral hemorrhage.

Objective To observe the changes of myelin sheath in corpus callosum on non-ischemic side after ischemic stroke in rats with cerebral ischemia/reperfusion(I/R)model,and to investigate the effect of vagus nerve stimulation(VNS)on secondary demyelination in I/R model.Methods Forty-two rats were randomly assigned to sham,I/R,or I/R+VNS groups,with 14 rats in each group.Except for the sham group,the rats in other groups were used to construct I/R model by a middle cerebral artery occlusion thread method.In addition,the rats in the I/R+VNS group were given repetitive electrical stimulation of the left vagus nerve for 60 min at 30 min after ischemia occlusion.The cerebral infarction volume was observed by 2,3,5-triphenyltetrazolium chloride staining.The expression levels of myelin basic protein(MBP)and myelin-associated glycoprotein(MAG)in the corpus callosum on the non-ischemic side were detected by Western blotting.The integrity and thickness of myelin sheaths in the corpus callosum on the non-ischemic side were observed by Luxol fast blue staining and transmission electron microscopy,respectively.Results Compared with the sham group,the volume of cerebral infarction in the I/R group was increased(P＜0.01).On the 3rd day after I/R,compared with the sham group,the expression levels of MBP and MAG were significantly decreased(both P＜0.01),the intensity of Luxol fast blue staining was significantly decreased(P＜0.01),and the thickness of the myelin sheath was significantly thinner(g-ratio was increased,P＜0.01)in the corpus callosum on the non-ischemic side.Compared with the I/R group,the expression levels of MBP and MAG in the I/R+VNS group were significantly increased(both P＜0.05),the intensity of Luxol fast blue staining was significantly increased(P＜0.01),and the thickness of myelin sheath was significantly increased(g-ratio was decreased,P＜0.01)in the corpus callosum on the non-ischemic side.Conclusion Secondary demyelination occurs on the non-ischemic side of the rat corpus callosum after ischemic stroke,and VNS can ameliorate the demyelination.

Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

This study investigated the effects and underlying mechanisms of the luteolin-naringenin combination(LN)on liver injury induced by acetaminophen(APAP).Forty-eight Kunming mice were randomly allocated into six groups:a normal control group,an APAP-induced liver injury model group,a positive drug treatment group,and three LN treatment groups with low,medium,and high doses.After the final drug administration,the mice were fasted for 12 hours prior to eu-thanasia for sample collection.Serum transaminase activity,oxidative stress indices,and hematoxy-lin-eosin(HE)staining were assessed to evaluate the effects of LN on APAP-induced hepatic inju-ry.Additionally,Western blot analysis was conducted to examine the expression levels of sphingo-sine kinase 1(SPHK1)and endoplasmic reticulum stress(ERS)-related proteins,thereby elucida-ting the potential mechanisms by which LN mitigates APAP-induced liver injury.The results dem-onstrated that varying concentrations of LN effectively ameliorated serum aminotransferase activi-ty and oxidative stress levels induced by APAP in a dose-dependent manner.Histopathological ex-amination via HE staining revealed significant improvement in APAP-induced liver tissue injury following treatment with different concentrations of LN.Furthermore,Western blot analysis indi-cated that the protein expressions of SPHK1,CHOP,p-IRE1α,ATF6,p-PERK,p-eIF2α,and ATF4 were markedly reduced after administration of various concentrations of LN.The results demonstrate that LN exhibits a significant protective effect against APAP-induced liver injury by inhibiting the SPHK1-mediated aberrant expression of ERS-related molecules.This study high-lights the importance of targeting SPHK1 in the treatment of APAP liver injury and provides a no-vel therapeutic approach through the multi-target and multi-pathway combination of monomers.

Objective To explore the value of combined detection of serum mitogen-activated protein kinase 1(MAPK1)and lysyl oxidase-like protein 2(LOXL2)in early diagnosis of cervical cancer.Methods A total of 218 patients with cervical lesions were selected as study group(103 ca-ses in cervical cancer group,115 cases in benign tumor group).Additionally,100 patients with cer-vical intraepithelial neoplasia grade Ⅱ were selected as precancerous lesion group,and 79 healthy in-dividuals undergoing physical examinations during the same period were selected as control group.Se-rum levels of MAPK1 and LOXL2 were measured in each group.Pearson correlation analysis was used to evaluate the correlations of serum MAPK1 and LOXL2 levels in patients with cervical cancer.Logistic regression analysis was performed to screen influencing factors for the occurrence of cervical cancer.Receiver operating characteristic(ROC)curves were plotted to assess the diagnostic efficacy of serum MAPK1 and LOXL2 for cervical cancer.Results Serum MAPK1 and LOXL2 levels in the study group were higher than those in the precancerous lesion group and the control group,and those in the precancerous lesion group were higher than those in the control group,with statistically signif-icant differences(P＜0.05).The proportion of patients with high-risk human papillomavirus(HPV)infection and serum MAPK1 and LOXL2 levels in the cervical cancer group were higher than those in the benign tumor group,with statistically significant differences(P＜0.05).Serum MAPK1 and LOXL2 levels in patients with stage Ⅲ to Ⅳ cervical cancer were higher than those in patients with stage Ⅰ to Ⅱ cervical cancer,with statistically significant differences(P＜0.05).Pearson correlation analysis showed a positive correlation between serum MAPK1 and LOXL2 levels in patients with cervical cancer(r=0.468,P＜0.001).Logistic regression analysis showed that high-risk HPV infection,MAPK1 and LOXL2 were all influencing factors for the occurrence of cervi-cal cancer(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)for com-bined diagnosis of serum MAPK1 and LOXL2 was 0.911,which was significantly greater than the AUCs for individual diagnoses(0.848 and 0.843,respectively).Conclusion Serum MAPK1 and LOXL2 levels in patients with cervical cancer are significantly upregulated,and the two indicators were positively correlated.High-risk HPV infection,serum MAPK1 and LOXL2 levels were influen-cing factors for the occurrence of cervical cancer.Combined detection of MAPK1 and LOXL2 levels is expected to assist in the diagnosis of cervical cancer.

Objective To translate the Maternal Childbirth Fatigue Perception Questionnaire(MCFQ)into Chinese and examine its reliability and validity among women undergoing vaginal deliv-ery.Methods Following the Brislin translation model,the MCFQ was translated,back-translated,culturally adapted and pre-tested to develop a Chinese version.By using the convenience sampling method,574 parturients were selected for a questionnaire survey from January to April 2024,and the reliability and validity of the Chinese questionnaire were evaluated.Results The Chinese version of MCFQ consisted of 3 dimensions and 13 items,namely physical fatigue(7 items),perceived fatigue(3 items)and emotional fatigue(3 items).The item-level content validity index ranged from0.813 to 1.000,and the scale-level content validity index was 0.938.The overall Cronbach's α coefficient of the scale was 0.870,and the split-half reliability coefficient was 0.848.Confirmatory factor analysis demonstrated that the question naire had a good fit.Conclusion The Chinese version of MCFQ has good reliability and validity and can be used as an effective tool to assess the fatigue level of parturients during the delivery process.It is helpful for clinical medical staff to optimize the management of the la-bor process,improve the quality of delivery,and ensure the safety of mothers and infants.