ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss （RPL） by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus （Shoutai Wan， Juyuan Jian）-of traditional Chinese medicine （TCM） at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c （blank group） and DBA/2 （modeling group） mice separately. Pregnant mice in the modeling group were randomly grouped as follows： high/low-dose Shoutai Wan， high/low-dose Juyuan Jian， model （RPL）， and positive control （dydrogesterone）， with 10 mice in each group. Starting from the day after the detection of the vaginal plug， mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention， the following indicators were measured. ① Macroscopic evaluation： general conditions， uterine wet weight， embryo loss rate， four coagulation parameters ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， fibrinogen （FIB）， and thrombin time （TT）］， and peripheral blood estradiol （E₂） and progesterone （Pg） levels. The decidua with embryos was stained with hematoxylin-eosin （HE） and evaluated by transmission electron microscopy （TEM）. The expression of B-cell lymphoma-2 （Bcl-2）， vascular endothelial growth factor （VEGF）， angiotensin Ⅱ （AngⅡ）， matrix metalloproteinase-2 （MMP-2）， interleukin-6 （IL-6）， leukemia inhibitory factor （LIF）， CXC chemokine ligand 12 （CXCL12）， and microtubule-associated protein 1 light chain 3 homolog （LC3）Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level： The decidua with embryos from the blank， model， high-dose Shoutai Wan， and high-dose Juyuan Jian groups （6 mice per group， with 3 single-cell samples per group， totaling 24 mice） were analyzed by the BD Rhapsody™ platform， and the whole-cell atlas was drawn by uniform manifold approximation and projection （UMAP） dimensionality reduction clustering combined with the single-cell mouse cell atlas （scMCA）. The differentially expressed genes （DEGs） and cell interaction networks were analyzed via Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and CellChat， and the protein-protein interaction （PPI） map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells （natural killer cells， NK cells） from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group， the RPL group showed an increase in the embryo loss rate （P<0.01）， down-regulated expression of Bcl-2， LIF， MMP-2， and Vegf in the decidua with embryos （P<0.05）， up-regulated protein levels of CXCL-12， AngⅡ， and IL-6 （P<0.05）， blocked angiogenesis， apoptosis-inflammation imbalance， and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance， and Shoutai pill showed superior performance in restoring the E₂ level to the Pg level （P<0.05）. ② Single-cell transcriptome analysis indicated that compared with the blank group， the RPL group showed differences in multiple key cell populations such as decidual cells， trophoblast cells， endothelial cells， erythroblasts， NK cells， and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group， high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer （upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes） and macrophages （upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes） through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins， cell adhesion， and programmed cell death， thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells （up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes） and macrophages （up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes）， which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D， CDH5， GDF， and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 （STAT3） and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis （TWEAK） signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7， a decreased proportion of reparative dNK4， and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1， thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method （Shoutai Wan） has an advantage in regulating the phenotypes of unfolded protein， cell adhesion， and programmed cell death， and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method （Juyuan Jian） has an advantage in regulating epithelial-mesenchymal transition （EMT）， angiogenesis， and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.

The incidence of papillary thyroid microcarcinoma (PTMC) is steadily rising. Although conventional thyroid surgery techniques are well-established, they present challenges such as functional impairment and overtreatment. Cellular-level fluorescence-guided imaging technology (EndoSCell^®) enables precise intraoperative tumor localization and real-time identification of neurovascular structures via high-resolution imaging and targeted labeling. This technology significantly improves surgical precision, reduces the incidence of postoperative complications including recurrent laryngeal nerve injury and hypoparathyroidism, and strengthens standards for functional preservation. This article aimed to systematically explore recent advances in the application of EndoSCell^® for PTMC management, analyze its influence on the innovation of surgical approaches and the enhancement of functional preservation criteria, and discuss future directions, thereby providing a theoretical basis for clinical practice.

Objective To establish and verify a double-antibody sandwich ELISA method for detecting the content of varicellazoster virus(VZV) glycoprotein E(VZV-gE) antigen in recombinant zoster virus-like particle(VLP) vaccine, so as to provide key technical support for the optimization of production process and the establishment of quality standards.MethodsMultiple anti-VZV-gE monoclonal antibodies were screened by epitope binning analysis to select capture antibody and detection antibody, and the working concentrations of which were determined by chessboard titration. A double-antibody sandwich ELISA was established to detect the VZV-gE antigen content in the drug substance and drug product of the recombinant zoster VLP vaccine. The linear range, specificity, dilution linearity, accuracy, precision and limit of quantitation(LOQ) of the method were investigated. In addition, the VZV-gE antigen content in multiple batches of drug substance and drug product of vaccine was detected using the established method to investigate the consistency of VZV-gE antigen content.ResultsA double-antibody sandwich ELISA for detecting VZV-gE antigen content in the recombinant zoster VLP vaccine was successfully established using ab272686 monoclonal antibody as capture antibody and VZV-M545 monoclonal antibody as detection antibody. This method could specifically identify VZV-gE antigen and had stability-indicating characteristics. The curve of VZV-gEM reference standard fitted well with A_(450)in the range of 0. 781-100 ng/mL, with R~2= 1. The dilution linearity ranges for detecting drug substance and lyophilized drug product were 4 100-262 400 and 200-12 800, respectively.The recovery rates of accuracy verification were all within the range of 80%-120%. Both the coefficients of variation(CVs)in intra-and inter-assay precision verification were less than 20%, while the LOQs for detecting VZV-gE antigen in the drug substance and lyophilized drug product were both 2 ng/mL. The detection results of VZV-gE antigen content in four batches of drug substance and lyophilized drug product met the expectations and showed good consistency.ConclusionThe established double-antibody sandwich ELISA has good specificity, accuracy, precision and linearity, which is suitable for the detection of VZV-gE antigen content in the recombinant zoster VLP vaccine.

Objective: To investigate the associations between childhood obesity and performance of six minute walk test (6MWT), providing evidence for exercise tolerance assessment and exercise intervention strategies for children and adolescents. Methods: From March 2021 to December 2023, a cohort study was conducted among students recruited from a primary and secondary school in Chongqing, a total of 709 valid samples were included. The 6MWT was used to assess exercise tolerance, with vital signs measured before and after the test. Anthropometric indicators, including height, weight, and waist circumference, were measured using standardized procedures. Generalized additive models (GAM) and restricted cubic spline (RCS) regression were employed to analyze the nonlinear relationships between obesity related indicators and six minute walk distance (6MWD). Results: The mean 6MWD of participants was (602.59±70.73)m. GAM showed that after adjusting for confounding factors, body mass index (BMI) and weight had non linear relationships with 6MWD [effective degrees of freedom were 1.55 and 7.13 respectively], and overweight/obesity was associated with a decrease in 6MWD ( β =-18.65) (all P <0.01). Further RCS regression analysis showed that both BMI and weight showed an "inverted U shaped" non linear relationship with 6MWD in the overall population and sex stratified subgroups; the 6MWD of females was lower than that of males, and it showed a significant downward trend with the increase of BMI or weight (all P <0.05). Conclusion Body weight and BMI in children and adolescents have an important impact on 6MWD, and obesity in children and adolescents is markedly associated with decline in exercise tolerance.

At present， the systematic excavation of the clinical experience and academic thought of the Sichuan school of Chinese medicine vis-à-vis its dominant disease entities remains fragmentary， and replicable paradigms are scarce. Confronted with empirical fragmentation， data heterogeneity and decision-making subjectivity， the standardised distillation， inheritance and clinical translation of these distinctive experiences has become a critical bottleneck constraining the development of the Sichuan school. The integration of artificial-intelligence technologies in data processing， pattern recognition and intelligent decision-making has rendered deep mining of traditional Chinese medicine（TCM） clinical knowledge and patterns imperative. Constructing an intelligent modern TCM diagnostic-therapeutic-evaluative system is now the obligatory route for inheritance and innovation in Chinese medicine， and simultaneously provides a technological breakthrough for intelligent decision paradigms in the dominant diseases of the Sichuan school. Accordingly， this study adopts the regional academic school as its point of entry， focuses on the dominant diseases of the Sichuan school， and proposes an innovative pathway of "four-dimensional data-multi-modal fusion-multi-agent decision-making". Specifically， four data dimensions are defined and instantiated： （Ⅰ） knowledge from classical medical literature and historical case records. （Ⅱ） objective four-diagnosis phenotypic data. （Ⅲ） master physicians' prescribing regularities. （Ⅳ） characteristic mechanisms of renowned formulae. Leveraging multi-modal data fusion and generative artificial intelligence， the entire causal chain of Famous Physicians and Renowned Formulas is explicated to reconstruct the diagnostic-therapeutic cognitive logic of the regional school. Finally， a multi-agent collective-decision model is established and refined for the dominant diseases of the Sichuan school， capable of generating precise， individualised treatment regimens and thereby advancing an intelligent diagnostic-therapeutic paradigm that delivers more efficient and accurate clinical decision support.

ObjectiveThis paper aims to conduct the feature analysis and correlation analysis on the ocular collateral features and differential metabolites in patients with chronic hepatitis B （CHB） complicated by glucose metabolism disorder （GMD），particularly those with the damp-heat syndrome type，by integrating shadowless scleral imaging and metabolomics technologies. MethodsA total of 313 patients were recruited from the Hepatology and Endocrinology Outpatient Departments of Public Health Clinical Center of Chengdu according to the inclusion/exclusion criteria，and they were divided into a CHB group and a CHB complicated by GMD groups （damp-heat syndrome group and non-damp-heat syndrome group）. All patients underwent high-definition ocular image acquisition and feature extraction using an intelligent analysis system for shadowless scleral imaging to analyze the differences in the counting of morphological feature scores of ocular collaterals among groups. By using a digital sampling method，24 patients from each group were randomly selected，along with 20 healthy volunteers，for untargeted metabolomic analysis of peripheral serum. Differential metabolites were identified，statistically analyzed，and subjected to potential biomarker analysis and pathway enrichment. Spearman method was performed to conduct the correlation analysis on the differential ocular collateral features and differential metabolites，followed by correlation network construction. ResultsCompared with those in the CHB group，patients with CHB complicated by GMD showed significant changes in ocular collateral feature scores such as "hillock"，"blood vessels"，and "pale dusky coloration" （P<0.05）. In comparison with those in the healthy group，metabolites including N-acetylglucosamine，acetylhomoserine，and myo-inositol （AUC>0.7） were identified as potential biomarkers for the disease. Compared with those in the CHB complicated by GMD group with non-damp-heat syndrome，patients with damp-heat syndrome exhibited significant changes in feature scores of "plaques"，"yellow coloration"，"spleen"，and "gallbladder" （P<0.05）. In comparison with those in the healthy group，metabolites such as O²′-4a-cyclic tetrahydrobiopterin，theobromine，xanthurenic acid，and L-glutamic acid 5-phosphate （AUC>0.7） were identified as potential biomarkers for the damp-heat syndrome type. The Spearman correlation analysis reveals weak to moderate linear correlations between the differential scleral collateral features and metabolites. By constructing a "disease-syndrome" network of ocular diagnosis and metabolites，"xanthurenic acid-gallbladder" and "theobromine-plaque/yellow coloration" were identified as specific molecular-phenotypic correlated biomarker clusters for CHB complicated by GMD with dampness-heat syndrome. ConclusionPatients with CHB complicated by GMD demonstrate differential ocular diagnostic features and serum metabolites corresponding to disease states and dampness-heat syndrome. These objective biomarkers can guide both clinical syndrome differentiation and medication. The macro-micro integration based on ocular feature clusters and potential metabolic biomarkers offers an innovative approach to a combined traditional Chinese and Western medicine diagnosis and treatment model for this disease.

Background/Aims: Chronic constipation is prevalent yet under-diagnosed across Asia, compromising quality of life and burdening healthcare systems. Cultural stigma, varied diets, and limited access to standardized diagnostic tools delay timely care. Methods: The Bali Chronic Constipation Roundtable in November 2024, brought together experts from 11 Asian countries. The group reviewed epidemiological data, analyzed multinational questionnaire on clinical practice pattern, and conducted structured discussions to identify key barriers and propose region-specific recommendations. Results: Chronic constipation prevalence varies across Asia, ranging from 1.8% in India to 16.6% in Japan, with women and the elderly disproportionately affected. Under-reporting persists owing to cultural taboos and widespread self treatment with laxatives and traditional medications. Although the Rome IV criteria remains the global standard, they may not fully reflect Asian symptom profiles, and diagnosis is limited by scarce motility laboratories. First line therapies such as dietary-fiber optimization and osmotic laxatives are widely available, but newer pharmacotherapies (prucalopride, linaclotide, lubiprostone, and elobixibat) remain costly and unevenly accessible. Biofeedback for dyssynergic defecation is underutilized due to limited availability. Experts recommend expanded regional research on to refine diagnostic criteria, coupled with enhanced physician education and public awareness. They advocate accessibility to second-line and novel therapies that incorporate culturally attuned regional guidelines, and improved access to gastrointestinal motility testing. Conclusions The Bali Chronic Constipation Roundtable highlighted Asia’s need for region specific diagnostics and management. Addressing diagnostic and treatment gaps will improve outcomes, while ongoing researcher clinician policy collaboration must standardize guidelines, advance research, and ensure equitable care across Asia.

Objective: To analyze changes in Rh system antibodies among antibody-positive patients and evaluate the efficacy of Rh phenotype-matched electronic cross-matching (hereinafter referred to as Rh-ECM). Methods: A retrospective analysis was performed on antibody screening data of 48 254 patients in our hospital from December 2023 to March 2025. The antibody screening results were compared between the pre-application phase (n=46 346, control group) and post-application phase (n=48 254, experimental group) of Rh-ECM technology, focusing on the changes in the proportion of Rh system antibodies, with statistical analysis conducted using SPSS 26.0 software. Meanwhile, the initial and re-examination situations of Rh antibody in the antibody screening of approximately 20 000 person-times each before (June 2019 to June 2020, n=21 048) and after (July 2020 to April 2021, n=20 965) of Rh-ECM were evaluated to explore the influence of Rh-ECM on the detection rate of Rh antibody. Results: After Rh-ECM implementation, 345 positive cases (0.7%) (345/48 254) were detected among 48 254 patients, primarily consisting of mns system antibodies (128 cases, 37.1%) (128/345) and rh system antibodies (95 cases, 27.5%) (95/345). Before Rh-ECM implementation, 199 positive cases (0.4%) (199/46 346) were detected among 46 346 patients, with rh system antibodies accounting for 97 cases (48.7%) (97/199). The difference in the composition ratio of Rh antibodies between the two phases was statistically significant (P<0.001), and the relative risk ratio of Rh antibody detection after Rh-ECM implementation was 56.5% compared to before. Another set of data analysis showed that before Rh-ECM, there were 37 cases with initial positive results and 8 cases with re-examination positive results; after Rh-ECM, these numbers were 44 and 2 respectively There was a statistically significant difference in the re-examination positive rate of Rh antibodies between the two stages (P<0.05). Conclusion: The implementation of Rh-ECM technology significantly reduced the proportion of Rh system antibodies among patients with positive antibody screening results. This suggests that Rh-ECM can effectively reduce the detection rate of Rh antibodies, which may be related to the reduced risk of antibody production due to Rh-matched transfusion, thus improving transfusion safety. Therefore, Rh-ECM is worthy of broader promotion in clinical transfusion testing.

Objective To investigate the changes of residual radioactivity at different time points after 131I treatment in the patients with differentiated thyroid cancer(DTC)and influencing factors.Methods A to-tal of 235 patients with DTC receiving 131I treatment in this hospital from January 2021 to June 2023 were se-lected as the study subjects and divided into the high dose group(＞5.55 GBp,n=56)and low dose group(≤5.55 GBp,n=179)according to the treatment dose.The clinical data of the two groups were collected and the changes of residual radioactivity after 131I treatment were compared between the two groups.The binary re-gression was used to analyze its influencing factors.Results The sex,age,BMI,basic metabolic rate(BMR)and serum thyroglobulin antibody(TgAb)had no statistical differences between the two groups(P＞0.05).The proportions of serum thyroglobulin(TG)＜1 ng/mL,131I first time treatment and residual thyroid ratio prompted by the whole body 131I scan after treatment in the low dose group were significantly higher than those in the high dose group(P＜0.05).The residual radioactivity in the two groups was significantly de-creased with time extension.The residual radioactivity at 24,48,72 h after treatment in the low dose group was significantly lower than that in the high dose group(P＜0.05).The binary logistic regression analysis re-sults showed that the T stage and treatment dose were the influencing factors of residual radioactivity after 131I treatment.Conclusion The residual radioactivity after 131I treatment in the patients with DTC shows the significant decreasing trend with time extension,this change trend has an active significance for further opti-mizing and perfecting the isolation and protection scheme.For the patients with high T stage and big treat-ment dose,the isolation time should exceed 72 h.

Objective To establish a human monocytic THP-1 macrophage-derived foam cell model and analyze the role of the α subunit of coatomer protein Ⅰ(α-COP)in apolipoprotein A-1(apoA-1)-mediated cholesterol efflux from foam cells.Methods THP-1 cells were induced to adhere using PMA(PMA group),followed by treatment with acetylated low-density lipoprotein(Ac-LDL)to generate macrophage-derived foam cells(Ac-LDL group).Subsequent incubation with apoA-1 formed the apoA-1 group.Cholesterol efflux rates mediated by apoA-1 and intracellular lipid accumulation were quantified through liquid scintillation counting and oil red O staining.The expression patterns of α-COP were systematically analyzed using quantitative real-time PCR(qPCR),Western blotting,and laser scanning confocal microscopy.THP-1 macrophage-derived foam cells mediated by apoA-1 were transduced with scrambled shRNA(Scr group)or α-COP-specific shRNA lentivirus(α-COP shRNA group)to assess the effects of α-COP knockdown on cholesterol efflux efficiency and total cellular cholesterol content.Subcellular localization of adipopoilin(ADFP)and cholesterol probe Fil-ipin Ⅲ staining patterns were visualized via laser scanning confocal microscopy.Bioinformatics analysis of α-COP expression profiles in carotid atherosclerotic plaques was performed using datasets from the GEO data-base.Results Compared with the Ac-LDL group,the cholesterol efflux rate in the apoA-1 group was signifi-cantly increased[(9.77±0.79)%vs.(2.74±0.37)%,P＜0.001].Oil red O staining demonstrated reduced lipid accumulation in foam cells of the apoA-1 group compared with the Ac-LDL group.The relative mRNA expression level of α-COP in the apoA-1 group was significantly higher than that in the Ac-LDL group(P＜0.001),with corresponding elevation in protein expression(P＜0.001).Fluorescence intensity analysis re-vealed increased mean fluorescence intensity of α-COP in the apoA-1 group compared with the Ac-LDL group.Following α-COP knockdown by shRNA,the apoA-1-mediated cholesterol efflux rate was significantly de-creased(P＜0.05)and total intracellular cholesterol levels were increased(P＜0.001)in the α-COP shRNA group compared with the Scr group.The α-COP shRNA group exhibited enhanced red fluorescence intensity of ADFP(P＜0.05)and increased blue fluorescence intensity of Filipin Ⅲ(P＜0.01)compared with the Scr group.Database analysis indicated that α-COP mRNA expression was lower in carotid atherosclerotic plaques than in normal arterial tissues,with reduced expression in advanced-stage plaques compared with early-stage plaques(P＜0.05).Conclusion α-COP participates in apoA-1-mediated cholesterol efflux from foam cells,and interference with α-COP expression results in reduced cholesterol efflux and increased intracellular lipid accumulation.