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.

Objective This study aimed to investigate the association between long-term radon exposure and changes in peripheral blood DNA methylation levels, as well as alterations in the expression of methylation-related regulatory genes (DNMT1, DNMT3A, and MBD2) among underground non-uranium metal miners. Methods Five non-uranium metal mines with varying levels of radon exposure were enrolled in this study. From each mine, 30 underground miners and 30 aboveground workers were randomly recruited. Basic demographic information was collected via a standardized questionnaire, and peripheral venous blood samples were obtained from all participants. An enzyme-linked immunosorbent assay (ELISA) colorimetric method was used to detect peripheral blood DNA methylation levels. Real-time quantitative PCR was used to measure mRNA expression levels of DNMT1, DNMT3A, and MBD2. These indicators were compared between underground and aboveground groups and among underground miners with different radon exposure levels. Multiple linear regression analysis was conducted to examine the associations between cumulative radon dose and two outcomes: peripheral blood DNA methylation levels, and the expression levels of methylation-related regulatory genes in underground miners. Results The average radon concentration in the underground workplaces of the five mines was 4,088 Bq/m³ (range: 116-18,188 Bq/m³), corresponding to an estimated annual effective dose of approximately 25.67 mSv for miners. The average radon concentration in aboveground workplaces was 58 Bq/m³ (range: 28-1796 Bq/m³), equivalent to an annual effective dose of approximately 0.51 mSv for aboveground workers. The average cumulative radon dose of underground miners was approximately 26.56 times that of aboveground workers (Z=−12.469, P<0.001). Compared with the aboveground workers, the peripheral blood DNA methylation level of underground miners was significantly decreased, approximately 0.77 times that of the aboveground group (Z=−2.313，P<0.05), accompanied by decreased expression of DNA methyltransferase genes DNMT1 and DNMT3A, which were 0.83 and 0.68 times those of the aboveground group, respectively (Z=−3.002、−4.190，P<0.01). After adjusting for age, working years, body mass index, smoking status, and alcohol consumption, multiple linear regression analysis revealed two key findings: cumulative radon dose was significantly and inversely correlated with peripheral blood DNA methylation levels (β=−0.752, P<0.001); and cumulative radon dose was significantly negatively associated with the expression of DNMT1 and DNMT3A (β=−0.319, −0.369, P<0.01). Conclusion The reduced DNA methylation levels and downregulated expression of DNA methyltransferase genes in underground miners are closely associated with long-term radon exposure.

Porcine deltacoronavirus(PDCoV)is the main pathogen of porcine deltacoronavirus dis-ease.After infection,pigsmanifest a series of main symptoms,such as persistent vomiting,watery diarrhea and severe dehydration.Pigs at almost all growth stages are likely to be infected with the virus,especially suckling piglets are much sensitive to the virus.Once PDCoV infects the host,it u-sually causes significant immunosuppression.In recent years,studies on the immunosuppressive mechanism of PDCoV have gradually attracted widespread attention.The results showed that mul-tiple proteins of PDCoV were involved in the regulation of host innate immunity,revealing the mechanism of these proteins in regulating host innate immunity.In this paper,the interaction mechanism between PDCoV protein and host innate immunity were rsummarized,which will pro-vide a theoretical basis for further understanding the pathogenesis of PDCoV and effective preven-tion and control of porcine delta coronavirus disease.

To establish a TaqMan-based multiplex RT-qPCR method for the identification of Japa-nese encephalitis virus(JEV),Porcine reproductive and respiratory syndrome virus(PRRSV),and Classical swine fever virus(CSFV),this study designed and synthesized three pairs of specific primers and probes based on the conserved sequences of JEV E,PRRSV ORF6,and CSFV E2 a-vailable in the NCBI GenBank.By optimizing the reaction system and protocol,a multiplex RT-qPCR method for detecting these three viruses was developed and applied to the detection of clini-cal samples.The results showed that the established TaqMan multiplex RT-qPCR specifically am-plified the gene fragments of JEV,PRRSV,and CSFV,and did not amplify other non-target genes,indicating good specificity of the method.Intra-assay and inter-assay repeatability tests showed that the coefficient of variation(Cv)values were all below 3％,demonstrating that the method has ex-cellent repeatability.Sensitivity tests revealed that the minimum detectable amount for the recom-binant plasmids of the three viruses was 100 copies/pL.Using the established method,a total of 969 samples,including blood,aborted fetuses,semen,and deceased pigs,from 26 pig farms in Guizhou Province were tested.The detection rates were 34.3％(332/969)for JEV,28.3％(274/969)for PRRSV,and 19.8％(192/969)for CSFV.The co-infection rates were 10.1％(98/969)for JEV and PRRSV,12.1％(117/969)for JEV and CSFV,and 14.6％(141/969)for CSFV and PRRSV.Additionally,the triple co-infection rate of JEV,PRRSV,and CSFV was 7.9％(77/969).These results indicate that the TaqMan multiplex RT-qPCR method developed in this study is ef-fective for detecting these three viruses in pig farms,providing technical support for identifying vi-ral causes of reproductive disorders.

Objective To investigate the mechanical effects of various height combinations of canine long-arm hooks and micro-im-plant anchorage on the retraction of maxillary anterior teeth in clear aligner treatment.It focuses on analyzing the stress distribution within the periodontal ligament and the movement tendencies of the anterior teeth,providing scientific evidence for optimizing orthodon-tic treatment strategies.Methods Three-dimensional finite element models were developed using CBCT scan data of patients and soft-ware including Mimics,Geomagic Studio,and Hypermesh.The models incorporated 16 different configurations of canine long-arm hooks(4,6,8,10 mm)and micro-implant anchorage(6,8,10,12 mm).Elastic traction forces of 3/16,3.5 oz were applied to these models.Simulations assessed how these variables influenced periodontal ligament stress distribution and the movement patterns of the maxillary anterior teeth.Results Stress during anterior teeth retraction was predominantly concentrated in the cervical and apex re-gions of the periodontal ligament,where it interfaced with the alveolar bone,indicating significant local concentration.Increasing the height of micro-implant anchorage and traction hooks markedly reduced positional changes in both the crown and apex.Specifically,when hooks exceeded 8 mm and anchorage heights surpassed 10 mm,canine movement became more uniform,preventing extrusion and minimizing the deepening of the overbite.Additionally,combinations of high traction hooks and high micro-implant anchorage promoted more uniform tooth movement,reducing unnecessary tipping and rotation.Conclusion In the retraction of maxillary anterior teeth with clear aligners,using high-position micro-implant anchorage and high-position traction hooks significantly enhances torque control of anterior teeth and effectively prevents deepening of the overbite.

Objective:To investigate the correlation of combined detection of p16 and Rb with high-risk human papilloma virus (HR-HPV) infection in non-oropharyngeal squamous cell carcinoma (NOPSCC) of the head and neck.Methods:A total of 68 NOPSCC cases of the head and neck (23 cases of the nasal cavity and paranasal sinuses and 45 cases of larynx) with complete clinical and pathological data, diagnosed at the Beijing Tongren Hospital, Capital Medical University, Beijing, China from November 2013 to December 2023, were collected. The expression of p16 and Rb was detected using immunohistochemistry of the EnVision two-step method, while the HR-HPV mRNA expression was detected using in situ hybridization. The concordance, sensitivity, and specificity of p16 alone and the combined detection of p16 and Rb for detecting HR-HPV infection were analyzed.Results:Among the 68 patients with NOPSCC, 53 were male and 15 were female, with a median age of 63.5 (range, 57.3 to 66.8) years. 41 patients had a smoking history and 27 did not. 33 patients had an early T stage (T1/T2) and 35 had advanced T stage (T3/T4). 14 patients had lymph node metastasis and 2 had distant metastasis. Histological types included 62 cases of keratinized squamous cell carcinoma, 5 cases of non-keratinized squamous cell carcinoma, and 1 case of basal-like squamous cell carcinoma. 25 cases were positive for p16. Among the 25 cases, 16 cases were positive for Rb, and 6 cases were positive for HR-HPV mRNA. 43 cases were negative for p16, including 38 cases positive for Rb and no cases positive for HR-HPV mRNA. The concordance between p16 and HR-HPV mRNA expression was poor ( Kappa=0.285, P=0.001), with a sensitivity of 100.0% and specificity of 69.4%. In contrast, the combined detection of p16+/Rb- showed high concordance with HR-HPV mRNA expression ( Kappa=0.719, P<0.001), with a sensitivity of 100.0% and specificity of 95.2%. Conclusions:In NOPSCC of the head and neck, the combined detection of p16 and Rb may be used as a marker for assessing HR-HPV infection. Recognizing the p16+/Rb- expression pattern in NOPSCC can improve the specificity of HR-HPV detection.

Objective Jiedu Tongluo Tiaogan Formula(JTTF)is an effective formula for the clinical treatment of type 2 diabetes mellitus(T2DM).We used integrated pharmacology and in vitro experiments to explore the molecular mechanism of JTTF to improve insulin resistance(IR)in T2DM.Methods The drug targets of JTTF were obtained by identifying the key active ingredients of JTTF through UPLC-Q-TOF-MS.Multiple databases such as GeneCards,OMIM,and DrugBank were used to screen T2DM-IR related targets.Cytoscape software and String 11.0 database were used to construct the PPI network diagram of JTTF for T2DM-IR.GO and KEGG analyses were performed according to the Metascape platform to find the biological pathways related to the target proteins.AutoDock Tools software was used to simulate molecular docking.In vitro experiments were performed using palmitic acid(PA)-induced HepG2-IR cell model to detect the effect of JTTF on HepG2-IR.Results 28 effective active components of JTTF were screened.There were 857 gene targets of T2DM-IR,and 168 targets of drug-disease intersection.387 GO entries and 145 KEGG pathways were enriched.The molecular docking results showed that the main components of JTTF had good binding activities with PI3K and AKT-related proteins.The in vitro results showed that JTTF significantly alleviated PA-induced HepG2 cell injury,increased HepG2 glucose consumption,increased PI3K and AKT mRNA and protein expression,regulated the expression of GLUT2,GLUT4 and GSK3β,and improved cellular IR.Conclusion JTTF increases insulin sensitivity of HepG2-IR cells,promotes glucose uptake and intracellular glucose metabolism process,and its mechanism of action may be related to the up-regulation of PI3K/AKT signalling pathway.

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.

AIM To study the chemical constituents from Tetrastigma hemsleyanum Diels et Gilg and their antitumor activity in vitro.METHODS Silica gel,ODS,Sephadex LH-20 and semi-preparative HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antitumor activity in vitro was determined by MTT mothod.RESULTS Twenty-eight compounds were isolated and identified as triphyllin A(1),eruberin B(2),(2S,4R)-5,7-dihydroxy-4,4'-dimethyl-6,8-dimethyl-flavan-5-O-β-D-6-acetylglucopyranoside-7-O-β-D-glucopyranoside(3),eruberin A(4),abacopterin Ⅰ(5),matteucinol(6),homoerodictyol(7),(2S)-5,3',4'-trihydroxy-7-methoxy-flavanone(8),(2S)-5,2',5'-trihydroxy-7-methoxyflavanone(9),galinsonside B(10),quercetin-3-O-β-D-glucopyranoside(11),kaempferol 3-O-robinobioside(12),rutin(13),geniposide(14),jasminoside A(15),β-sitostenone(16),sitosterol palmitate(17),β-sitosterol(18),ursolic acid(19),hyptadienic acid(20),3,4-dihydroxybenzoic acid(21),3,4-dimethoxybenzoic acid(22),gallic acid(23),dibutylphthalate(24),bis-(2-ethylhexyl)phthalate(25),9-nonadecenoic acid(26),triacylglycerol(27),crocin Ⅰ(28).The IC50 values of compound 1 for human gastric adenocarcinoma cells BGC-823 and human colon cancer cells HCT-116 were(22.07±0.38),(20.67±0.11)μmol/L,respectively.The IC50 value of compound 9 for BGC-823 cells was(21.58±0.05)μmol/L,and the IC50 value of compound 4 for HCT-116 cells was(16.67±0.36)μmol/L.CONCLUSION Compounds 1-10,14-15 and 28 are first isolated from Tetrastigma genus.Compounds 1,4,9 have weak antitumor activity in vitro.

Distal transradial access(dTRA)was first applied in coronary heart disease intervention by Kiemeneij in 2017,dTRA has become an important technological advancement of coronary interventional therapy.This approach,performed by puncturing the distal radial artery within the anatomical snuffbox or Hegu point,significantly reduces the incidence of radial artery occlusion compared with conventional transradial access and shortens compression time to 2-3 hours after procedure.Clinical evidence confirms the efficacy of dTRA in facilitating complex percutaneous coronary interventions,including stenting of left main coronary artery bifurcation lesions and recanalization of chronic total occlusions.Its application has progressively expanded to neurointerventional procedures(cerebral angiography),tumor embolization(transarterial chemoembolization for hepatocellular carcinoma),and peripheral vascular interventions.Despite increasing clinical adoption,dTRA still faces challenges,including a long learning curve and relatively lower initial puncture success rates.Combined with the current paucity of robust evidence-based data,the viability of dTRA as a routine interventional access route remains a subject of debate.This article systematically reviews the anatomical landmarks,clinical advantages,limitations,and multidisciplinary applications of dTRA.This article aims to provide practical guidance for interventionalists and promote the standardization of this technique in daily clinical practice.