This paper summarizes Professor LU Fang's clinical experience in treating systemic lupus erythematosus （SLE） based on the differentiation and treatment of heat-toxin and blood-stasis in the collaterals. SLE is generally characterized by deficiency in origin with excess in manifestation. The core pathogenesis is heat-toxin obstructing the collaterals. During the acute active stage， the predominant pattern is blazing heat-toxin causing blood stasis， while in the chronic remitting stage， the main pattern is toxic stasis blocking the collaterals with qi and yin deficiency. Clinical treatment follows the basic principle that treat with salty-cold herbs， when heat invades internally and that assist with acrid-dispersing herbs when stasis obstructs the collaterals. The self-formulated Yimian Decoction （抑免汤） serves as the base formula and is applied in stages. During the acute active stage， it is often combined with herbs for clearing heat and detoxifying， cooling blood and resolving stasis， and unblocking the collaterals. In the chronic remitting stage， it is often combined with herbs for activating blood circulation and unblocking the collaterals， as well as tonifying qi and nourishing yin.

Japanese encephalitis virus(JEV)belongs to the genus Flavivirus and the family Flavi-viridae,and is classified as a single-stranded,positive-sense RNA virus.The disease known as Japa-nese encephalitis(JE),which results from JEV infection,is a viral zoonosis that is prevalent worldwide and poses a significant public health concern.JEV infection activates a variety of signa-ling pathways,leading to a series of changes that are crucial to the virus's pathogenesis.Among these pathways,Toll-like receptors(TLRs)are particularly significant,and their diverse range and complex signal transduction mechanisms present substantial challenges for the prevention and con-trol of JEV.Currently,there is no specific treatment for JEV.Although some vaccines have been developed to prevent JE,eradicating JEV remains difficult due to its zoonotic transmission cycle and the limited efficacy of the available vaccines.This article reviews the alterations in various TLR signaling pathways induced by JEV infection in the host,aiming to provide insights into the patho-genic mechanisms of JEV and to identify potential new antiviral targets.

Porcine reproductive and respiratory syndrome virus(PRRSV)is the pathogen of porcine reproductive and respiratory syndrome(PRRS),and its infection mainly causes abortion,stillbirth in sows and piglet respiratory infections,which are widely prevalent in the world and seriously jeopardize the development of the world's animal husbandry industry.PRRSV infection of the host is capable of inducing significant immunosuppression,and in recent years,the study of the mecha-nism of PRRSV immunosuppression has become a hot topic,with studies showing that numerous PRRSV proteins are involved in the regulation of host innate immunity and elucidating the mecha-nism by which PRRSV proteins modulate host innate immunity.In this paper,we reviewed the progress of research on the interaction mechanism between PRRSV proteins and host innate im-munity to provide a theoretical basis for a comprehensive understanding of the pathogenesis of PRRSV and the prevention and control of PRRS.

Objective To compare the ability of single-phase,dual-phase,and triphasic models in forecasting postoperative pancreatic fistula(POPF)after pancreaticoduodenectomy(PD)using radiomics based on triphasic enhanced CT.Methods A total of 181 patients who underwent multi-phase enhanced CT prior to PD were retrospectively selected,and the collection phase included non-contrast,arterial phase(AP),and equilibrium phase(EP).3D Slicer software was utilized to segment the region of interest(ROI)for the postoperative pancreatic remnant on each phase.Radiomics feature extraction was performed using R software,followed by feature selection through least absolute shrinkage and selection operator(LASSO)regression with five-fold cross-validation to prevent model overfitting.The effective features selected were combined in a weighted linear manner to obtain a Radiomics score(Radscore).The patients were divided into training set and test set in a 7︰3 ratio.Logistic regression was employed to construct seven POPF prediction models(three single-phase,three dual-phase,and one triphasic models)based on different phase combinations.The diagnostic performance of the models was evaluated using the area under the curve(AUC)of receiver operating characteristic(ROC)curve,accuracy(ACC),sensitivity(SEN),and specificity(SPE).The DeLong test was applied to compare the differences in AUC among different models.Results After LASSO regression,24 effective features associated with POPF were selected from different phases.In the test set,the triphasic model exhibited the highest AUC and ACC(AUC=0.76,ACC=0.808).The calibration curve demonstrated the strongest agreement between the estimated probabilities and observed probabilities for the triphasic model.The decision curve analysis(DCA)curve indicated that the triphasic model had the largest threshold range with a higher net benefit.Conclusion Compared with single-phase and dual-phase models,the triphasic model based on enhanced CT provides better prediction of POPF after PD,aiding clinical decision-making and improve prognosis.

Objective To investigate the diagnostic and therapeutic value of single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis therapy in the treatment of acute uncomplicated appendicitis.Methods A retrospective analysis was conducted on the clinical data of 39 patients with acute uncomplicated appendicitis who underwent single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis treatment at the Endoscopy center of the hospital from September 2022 to September 2024.Observe the endoscopic manifestations,the rate of maternal and child basket stone removal,the rate of appendiceal stent implantation,the technical success rate,the clinical success rate,the operation time,the hospital stay,the incidence of complications,the visual analogue scale(VAS)score 6 hours after the operation,and the inflammatory indicators 24 hours after the operation.Results In 28 cases(71.8%),congestion and edema could be seen at the opening of the appendix under colonoscopy.In 10 cases(25.6%),pus could be seen flowing out of the opening of the appendix under colonoscopy.In 32 cases(82.1%),a large amount of pus could be seen in the lumen of the appendix under subscopy.In 20 cases(51.3%),appendiceal fecalith could be seen in the lumen of the appendix under subscopy.The technical success rate of single-use mother-baby choledochoscope-assisted endoscopic retrograde appendicitis treatment was 100.0%(39/39).The operation time was(21.08±7.49)min;Hospital stay:(3.97±2.08)days;Eight cases(20.5%)of patients underwent endoscopic maternal basket stone removal.Appendiceal stent implantation was performed in 14 cases(35.9%)of patients.The clinical success rate is 97.4%(38/39).One patient's clinical symptoms and inflammatory indicators did not improve after the operation,and was transferred to the surgery department for appendectomy.The VAS score of 38 patients was less than 3 points 6 hours after the operation,and the abdominal pain symptoms were significantly relieved.The white blood cell count and the percentage of neutrophils 24 hours after the operation decreased significantly compared with those before the operation,and the differences were statistically significant(P<0.05).None of the 39 patients had complications.The postoperative follow-up was(5.94±4.03)months,and recurrence occurred in 3 cases(7.7%).Conclusion single-use mother-baby choledocoscope-assisted endoscopic retrograde appendicitis therapy is safe and effective in the diagnosis and treatment of acute uncomplicated appendicitis,which is worthy of further promotion and popularization in clinical practice.

BACKGROUND:In vitro reconstruction of hair follicles depends on the self-assembly behavior of dermal papilla cells and keratinocytes during three dimensional co-culture,which is very similar to the development of hair follicle embryos.Therefore,hair follicle organoids can be used as a good model in vitro for basic research of hair follicle regeneration.Currently,wholly humanized follicle organoids cultured by biomimetic system have been successfully transplanted into animals.Research on hair follicle organoids makes it possible to regenerate hair follicles,but there is no uniform conclusion on the construction method,and there are still many challenges.OBJECTIVE:To review the current research progress of hair follicle organoids,summarize their culture methods and characterization,and expound their application prospects,so as to provide reliable basis for subsequent research,in order to promote its clinical transformation in tissue engineering and alopecia treatment.METHODS:CNKI and PubMed databases were searched by a computer,using"hair follicle regeneration,hair regeneration,organoid"as Chinese search terms and"hair growth,hair loss,hair regeneration,hair follicle,dermal papilla cell,organoid"as English search terms.The domestic and foreign literature on hair follicle organoids was reviewed and a total of 67 articles were included for follow-up analysis.RESULTS AND CONCLUSION:(1)Dermal papilla cells and keratinocytes from different sources can be used to construct hair follicle organoids in vitro to obtain mouse-derived,human-mouse chimeric and human-derived hair follicle organoids,which have been successfully used in rodent transplantation models.(2)A variety of programming methods,such as electrical stimulation,addition of recombinant protein,and dissociation of cell gene expression,can be used to promote hair follicle organoid such ashair follicle germ formation and hair shaft elongation.Simultaneously,the application of various 3D printing technologies has achieved controllable hair follicle organoid to the size of the quantity production,which greatly improves the production efficiency.(3)When melanocytes are added to the dissociated cell co-culture system,the problem of pigment deficiency in new hair follicles may be solved.When human umbilical vein endothelial cells are added,the new blood vessels in the cell aggregates can improve their nutrient supply and reduce necrosis in the central area.(4)As the technology of engineered hair follicle organoid gradually maturates,hair follicle organoids constructed from patient's autologous cells may be used for individualized treatment of severe alopecia.

Objective To investigate the impact of long non-coding RNA maternal expression gene 3(lncRNA MEG3)on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis by modulating the miR-202-5p/signal transducer and activator of transcription 3(STAT3)axis.Methods An H/R cell model was constructed and randomly separated into four groups:sh-NC(transfected with NC shRNA),sh-MEG3(transfected with lncRNA MEG3 shRNA),miR-NC(transfected with lncRNA MEG3 shRNA and NC miR),and in-miR-202-5p(transfected with lncRNA MEG3 shRNA and miR-202-5p inhibitor).In addition,the H/R group(nontransfected H/R model cells)and the AC 16 group(normal AC 16 cells)were set.Quantitative real-time PCR method was used to analyze the expression of lncRNA MEG3,miR-202-5p,and STAT3 in cells from each group.CCK-8 method was used to analyze cell viability.Flow cytometry was used to analyze apoptosis.Enzyme-linked immunosorbent,colorimetric,and probe assays were applied to detect the levels of inter-leukin-6(IL-6),tumor necrosis factor α(TNF-α),malondialdehyde(MD A),glutathione peroxidases(GSH-Px),and reactive oxygen spe-cies(ROS).Western blotting was carried out to examine the expression of STAT3,Bcl-2-associated X protein(Bax),B-cell lymphoma-2(Bcl-2),caspase-3,and caspase-9.A dual luciferase assay was used to analyze the relationship between lncRNA MEG3 and miR-202-5p,as well as the relationship between miR-202-5p and STAT3.Results Compared with that in the AC 16 group,the expression of lncRNA MEG3 and STAT3 in cells in the H/R group increased,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the expression of lncRNA MEG3 and STAT3 decreased in the sh-MEG3 group,while the expression of miR-202-5p increased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the expression of lncRNA MEG3 and STAT3 increased in the in-miR-202-5p group,while the expression of miR-202-5p decreased(P＜0.05).Compared with that in the AC 16 group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the H/R group.In contrast,the cell viability,clone count,levels of superoxide dismutase(SOD)and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).Compared with that in the H/R group and sh-NC group,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 increased in the sh-MEG3 group.In contrast,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 decreased(P＜0.05).Compared with that in the sh-MEG3 group and miR-NC group,the apoptosis rate,levels of IL-6,TNF-α,MDA,and ROS,and the expression of STAT3,Bax,cleaved caspase-3,and cleaved caspase-9 increased in the in-miR-202-5p group.In contrast,the cell viability,clone count,levels of SOD and GSH-Px,and the expression of Bcl-2 decreased(P＜0.05).There were multiple binding sites between lncRNA MEG3 and miR-202-5p,and between miR-202-5p and STAT3.The luciferase activity was lower in the WT-MEG3+miR-202-5p group than in the WT-MEG3+miR-NC group(P＜0.05).The luciferase activity was lower in the WT-STAT3+miR-202-5p group than in the WT-STAT3+miR-NC group(P＜0.05).Conclusion Knocking down lncRNA MEG3 can downregulate STAT3 by negatively regulating miR-202-5p,inhibiting H/R-induced cardiomyocyte apoptosis.

Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

Objective To explore the mechanism of Glaucocalyxin A(GLA)in inhibiting ovalbumin(OVA)-induced airway remodeling in asthmatic mice through the topoisomerase Ⅱ α(TOP2A)/cyclin-dependent kinase 1(CDK1)signaling pathway.Methods Forty Balb/c mice were randomly divided into 5 groups:the control group,the model group,the low-dose GLA group,the high-dose GLA group and the Dexamethasone group,with 8 mice in each group.The effect of GLA on airway remodeling was examined by immunohistochemical staining,ELISA and other methods,and bioinformatics methods were used to predict new targets of GLA.The action targets of TOP2A were screened using the STRING database,and the interaction relationship between the two was verified by co-immunoprecipitation.In vitro,GLA and siRNA were used to interfere with interleukin-4(IL-4)-stimulated human airway epithelial cells BEAS-2B.The expressions of TOP2A,epidermal growth factor receptor(EGFR),Integrin β1,focal adhesion kinase(FAK),β-catenin,CDK1 and DRP1 were detected by Western Blot.Results GLA intervention could significantly reduce OVA-induced asthma airway remodeling,airway smooth muscle thickening,collagen deposition around the airway,the number of eosinophils in alveolar lavage fluid,the expression of pro-inflammatory cytokines such as IL-4,and the level of serum IgE.The new target of GLA screened out was TOP2A,which was highly expressed in the lung tissue of the asthma airway remodeling model.GLA intervention could down-regulate its expression.In vitro,intervention with GLA and si-TOP2A could significantly down-regulate the expressions of IL-4-induced TOP2A,EGFR,Integrin β1,FAK and β-catenin.Further studies have found that TOP2A had an interaction relationship with CDK1.si-TOP2A could downregulate the expression of CDK1,and knockdown of CDK1 could significantly down-regulate the expression of phosphorylated DRP1.Conclusion GLA may alleviate asthma airway remodeling by targeting the TOP2A/CDK1 signaling pathway,providing experimental evidence for the clinical diagnosis and treatment of asthma airway remodeling in asthma.

To identify clinical viral diseases characterized by reproductive disorders and abortion,a quadruple qPCR method was established for simultaneous detection of PRV,PPV,PCV2 and AS-FV.Four pairs of specific primers and probes were designed according to the conserved genes of four viruses in the NCBI gene bank.The annealing temperature,primer concentration and probe concentration of the reaction were optimized,and the specificity,sensitivity and repeatability of the method were tested.The results showed that the method could not detect other pathogens except the target ones.The minimum detection limit of PRV,PPV,PCV2 and ASFV was 10 copies.Intra-group and inter-group repeatability tests showed that the coefficient of variation of C,values be-tween different batches was less than 3％,indicating that the method was highly specific,sensitive and stable.Establishment of an efficient and sensitive quadruple qPCR method provides technical reference for the clinical prevention and control of porcine pseudorabies virus disease,porcine circo-virus disease,porcine parvovirus disease and African swine fever.