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.

BACKGROUND:The use of 3D-printed patient-specific instruments in opening wedge high tibial osteotomy has advantages such as shorter operative time,fewer fluoroscopic exposures,and higher correction accuracy.However,previous studies have reported issues such as significant damage to surrounding soft tissues and improper fixation of the plates. OBJECTIVE:To investigate the clinical efficacy of using 3D-printed patient-specific instruments combined with customized locking plate in opening wedge high tibial osteotomy for the treatment of knee osteoarthritis. METHODS:A total of 20 patients diagnosed with knee osteoarthritis were divided into the 3D group(n=10)and the conventional group(n=10)according to surgical methods.The 3D group underwent opening wedge high tibial osteotomy using 3D-printed patient-specific instruments combined with customized locking plate,while the conventional group underwent opening wedge high tibial osteotomy using conventional methods.The operative time,fluoroscopic exposures,incision length,pre-and postoperative hip-knee-ankle angle,medial proximal tibial angle,posterior tibial slope,the difference between the planned and actual correction angle,preoperative and 1,3,6 months postoperative knee range of motion and Lysholm score,and incidence of complications were analyzed and compared between the two groups. RESULTS AND CONCLUSION:(1)The operative time and fluoroscopic exposures were significantly shorter in the 3D group compared to the conventional group,with a statistically significant difference(P<0.001).(2)Both groups showed a significant improvement in postoperative hip-knee-ankle angle and medial proximal tibial angle compared to preoperative values,with a statistically significant difference(P<0.001),while there was no significant change in posterior tibial slope.In the 3D group,the postoperative hip-knee-ankle angle,medial proximal tibial angle,and posterior tibial slope differed from their respective preoperative planned values by(-0.22±0.72)°,(-0.20±0.73)°,and(0.23±0.37)°,but the differences were not statistically significant.The difference between the planned and actual correction angle of 3D group was significantly smaller than that of conventional group(P<0.05).(3)Both groups showed a gradual increase in knee range of motion and Lysholm scores after surgery(P<0.001).Compared to the conventional group,the 3D group had superior knee range of motion at 1 and 3 months postoperatively,as well as a higher Lysholm score at 1 month postoperatively,with statistically significant differences(P<0.05).There were no statistically significant differences in Lysholm score at 3 months and knee range of motion and Lysholm score at 6 months between the two groups(P>0.05).(4)Complications occurred in neither groups.(5)The above results indicate that both 3D-printed patient-specific instruments combined with customized locking plate and conventional methods have good clinical efficacy.However,the former has a shorter operative time,fewer fluoroscopic exposures,and faster postoperative recovery of knee joint function.Additionally,3D-printed patient-specific instruments can achieve preoperative planning accurately.

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.

This article presents a comprehensive review of new methods for target identification of complex components in tradi-tional Chinese medicine(TCM)and research progress in their applications.It systematically summarizes classical approaches and cutting-edge technical systems for target identification,cov-ering multiple strategies such as proteomic analysis,fluorescence resonance energy transfer(FRET)technology,network pharma-cology prediction models,high-throughput biochip screening,tar-get capture strategies based on molecular affinity fishing,gene site-directed mutagenesis verification,and co-crystallization structure analysis of target proteins.The review emphasizes the critical role of target identification in elucidating the action mechanisms of TCM and facilitating new drug development,lay-ing a foundation for promoting the modernization of TCM.

Objective:To investigate the molecular mechanism by which 2',4'-dihydroxychalcone(D2)inhibits proliferation,migration,and epithelial-mesenchymal transition(EMT)in colorectal cancer cells through miR-7-5p-mediated autophagy.Methods:Human colorectal cancer cell lines HCT-15 and SW620 were treated with D2 at concentrations of 0,12.5,25,and 50 μmol/L.Cell proliferation and clonogenic capacity were evaluated using MTT and colony formation assays.Cell migration was assessed by wound healing and Transwell assays.WB assay was used to detect the expression of EMT-related proteins,autophagy-related proteins,and key components of the PI3K/AKT/mTOR pathway.Autophagosome formation was visualized by immunofluorescence staining.TCGA database and KEGG pathway analyses were performed to evaluate miR-7-5p expression and its association with colorectal cancer.RT-qPCR was used to quantify miR-7-5p expression,and lentiviral transduction was employed to establish stable miR-7-5p knockdown or overexpression cell lines.Results:D2 significantly inhibited colorectal cancer cell proliferation,migration,and EMT(P<0.05 or P<0.01).TCGA and KEGG analyses revealed that miR-7-5p expression was downregulated in colorectal cancer and closely associated with disease progression.D2 treatment(12.5,25,and 50 μmol/L)significantly upregulated miR-7-5p expression in HCT-15 and SW620 cells(P<0.01).At 25 μmol/L,D2 increased the expression of autophagy-related proteins(LC3 and p-ULK1)and inhibited the PI3K/AKT/mTOR signaling pathway(P<0.05),accompanied by increased autophagosome formation(P<0.01).In miR-7-5p-knockdown cells treated with D2,the levels of LC3 and p-ULK1 were significantly reduced compared to D2-only treated cells(P<0.05 or P<0.01).Conclusion:D2 upregulates miR-7-5p to induce autophagy,thereby inhibiting colorectal cancer cell proliferation,migration,and EMT,possibly through suppression of the PI3K/AKT/mTOR signaling pathway.

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.