1.Consistent Detection of Aquaporin-4 Antibodies:A Comparative Analysis Between Fixed and Live Cell-Based Assays
Jing WANG ; Linge WANG ; Xiaolin YANG ; Zhizhong LI ; Jinyu JIANG ; Qiao XU ; Siyuan HUANG ; Qing FU ; Yang YANG ; Rongrong ZHANG ; Lin YANG ; Ai CHEN ; Xiaopeng ZENG ; Ke XU ; Peng ZHENG ; Xinyue QIN ; Jinzhou FENG
Journal of Clinical Neurology 2026;22(2):212-220
Background:
and Purpose Live cell-based assays (LCBA) are increasingly used for serological antibody detection due to their ability to preserve antigen conformation, offering moderately higher sensitivity than fixed cell-based assays. However, the clinical necessity of prioritizing LCBA for the detection of aquaporin-4 immunoglobulin G (AQP4-IgG) in neuromyelitis optica spectrum disorder (NMOSD) remains unclear, especially when compared to its established role in diagnosing myelin oligodendrocyte glycoprotein antibody-associated disease.
Methods:
We compared the performance of live cell-based assays using immunofluorescence (LCBA-IF) and fixed cell-based assays using immunofluorescence (FCBA-IF) in detecting AQP4-IgG in 90 cases of NMOSD meeting 2015 International Panel for Neuromyelitis Optica Diagnosis criteria, alongside 40 controls. Additionally, we further investigated the relationship between AQP4-IgG titers as measured by LCBA-IF and FCBA-IF and clinical parameters in NMOSD patients.
Results:
Results showed 96.9% agreement between LCBA-IF and FCBA-IF (Cohen’s κ=0.935), with a strong Spearman correlation (0.977, p<0.001). Both methods demonstrated 100% specificity, with LCBA-IF showing slightly higher sensitivity compared to FCBA-IF. Within LCBAIF-tested groups, statistically significant differences in annualized relapse rates were observed across all pairwise comparisons (low-titer vs. moderate-titer, low-titer vs. high-titer, and moderate-titer vs. high-titer; all p<0.050). However, this association reached statistical significance in some FCBA-IF-tested groups.
Conclusions
Overall, there is a strong concordance between LCBA-IF and FCBA-IF in detecting AQP4-IgG, where LCBA-IF shows slightly higher sensitivity. Furthermore, there is a potential link between elevated AQP4-IgG titers and an increased risk of relapse, and this correlation may appear more pronounced when using LCBA-IF.
2.Quercetin Ameliorates Gouty Arthritis in Rats via ROS/NLRP3/IL-1β Signaling Pathway
Baowei FENG ; Yan WANG ; Chang LI ; Yujing ZHANG ; Dingxing FAN ; Xin LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):145-153
ObjectiveTo investigate the effect of quercetin on acute gouty arthritis (GA) in rats by inhibiting the reactive oxygen species (ROS)/NOD-like receptor protein 3 (NLRP3)/interleukin-1β (IL-1β) signaling pathway. MethodsSixty SPF-grade male SD rats were randomized into normal, model, colchicine (0.3 mg·kg-1), and low-, medium-, and high-dose (25, 50, 100 mg·kg-1, respectively) quercetin groups (n=10). The rats in the dosing groups were administrated with the corresponding drugs (10 mL·kg-1) by gavage once a day for one week. An equal volume of normal saline was given by gavage to rats in normal and model groups. One hour after drug administration on day 5, an acute GA model was established in other groups except the control group via intra-articular injection of monosodium urate (MSU) suspension into the right posterior ankle joint cavity. The joint swelling and gait were scored at the time points of 6, 12, 24, 48 h after modeling. Histopathological alterations in the ankle joint tissue from each group were assessed by hematoxylin-eosin (HE) staining. Malondialdehyde (MDA), xanthine oxidase (XOD), and total superoxide dismutase (T-SOD) assay kits were used to assess the levels of MDA, XOD, and T-SOD in the serum. The levels of tumor interleukin-6 (IL-6), necrosis factor-α (TNF-α), and IL-1β in the rat serum, as well as ROS in the ankle joint tissue, were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was performed to determine the protein levels of NLRP3, thioredoxin-interacting protein (TXNIP), apoptosis-associated speck-like protein containing a CARD domain (ASC), precursor cysteinyl aspartate-specific proteinase-1 (pro-Caspase-1), cleaved Caspase-1 (Caspase-1 p20), and IL-1β in the ankle joint tissue. Real-time PCR was employed to assess the mRNA levels of TXNIP, NLRP3, ASC, IL-1β, and TNF-α in the ankle joint tissue. ResultsCompared with the normal group, the model group exhibited decreased spontaneous activity, mental fatigue, increased ankle joint swelling and gait scores (P<0.01), aggravated synovial tissue edema and inflammatory cell infiltration (P<0.01), elevated levels of XOD, MDA, TNF-α, IL-1β, and IL-6 in the serum and ROS in the joint tissue (P<0.01), a declined level of T-SOD (P<0.01), up-regulated protein levels of NLRP3, TXNIP, ASC, pro-Caspase-1, Caspase-1 p20, and IL-1β in the ankle joint tissue (P<0.01), and up-regulated mRNA levels of NLRP3, TXNIP, ASC, IL-1β, and TNF-α in the ankle joint tissue (P<0.01). Compared with the model group, the medium- and high-dose quercetin groups showed improved general conditions, decreased gait scores (P<0.05, P<0.01), reduced joint swelling (P<0.01), alleviated synovial tissue edema and inflammatory cell infiltration (P<0.05, P<0.01), lowered levels of XOD, MDA, TNF-α, IL-1β, and IL-6 in the serum and ROS in the joint tissue (P<0.01), increased levels of T-SOD (P<0.01), down-regulated protein levels of TXNIP, NLRP3, ASC, pro-Caspase-1, Caspase-1 p20, and IL-1β in the ankle joint tissue (P<0.05, P<0.01), and down-regulated mRNA levels of TXNIP, NLRP3, ASC, IL-1β, and TNF-α in the ankle joint tissue (P<0.01). Low-dose quercetin also ameliorated some of the above parameters (P<0.05, P<0.01). ConclusionQuercetin exerts anti-GA effects by blocking the ROS/NLRP3/IL-1β signaling pathway, downregulating NLRP3 inflammasome activation, and inhibiting the production of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6.
3.Molecular Mechanism of Programmed Cell Death in Chronic Obstructive Pulmonary Disease and Traditional Chinese Medicine Intervention: A Review
Xin PENG ; Yunhui LI ; Lei LIANG ; Zheyu LUAN ; Hanxiao WANG ; Haotian XU ; Ziming DANG ; Jihong FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):304-313
Chronic obstructive pulmonary disease (COPD) is a chronic respiratory disease that poses a significant threat to global health, exhibiting high morbidity, disability and mortality rate, with its prevention and treatment situation becoming increasingly critical. The pathogenesis of COPD is complex, and the underlying cellular and molecular biological mechanisms remain incompletely elucidated. Programmed cell death (PCD) is the process wherein cells actively undergo demise to maintain internal environmental stability in response to certain signals or specific stimuli. Contemporary medical research indicates that the dysregulation of PCD patterns such as apoptosis, necroptosis, pyroptosis, autophagy, and ferroptosis is closely related to the onset and progression of COPD. Clarifying the molecular mechanisms of PCD in COPD may provide novel perspectives for in-depth understanding and prevention of the disease. Traditional Chinese medicine (TCM) is characterized by holistic regulation. In recent years, extensive research has been conducted in the TCM field focusing on modulating apoptosis, necroptosis, pyroptosis, autophagy, and ferroptosis for the treatment of COPD, yielding remarkable achievements. Therefore, this study systematically explored the molecular mechanism of PCD in COPD and reviewed the potential mechanisms and intervention status of TCM targeting PCD in COPD, aiming to provide insights and references for the clinical prevention, treatment and in-depth research of COPD.
4.Molecular Mechanism of Programmed Cell Death in Chronic Obstructive Pulmonary Disease and Traditional Chinese Medicine Intervention: A Review
Xin PENG ; Yunhui LI ; Lei LIANG ; Zheyu LUAN ; Hanxiao WANG ; Haotian XU ; Ziming DANG ; Jihong FENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):304-313
Chronic obstructive pulmonary disease (COPD) is a chronic respiratory disease that poses a significant threat to global health, exhibiting high morbidity, disability and mortality rate, with its prevention and treatment situation becoming increasingly critical. The pathogenesis of COPD is complex, and the underlying cellular and molecular biological mechanisms remain incompletely elucidated. Programmed cell death (PCD) is the process wherein cells actively undergo demise to maintain internal environmental stability in response to certain signals or specific stimuli. Contemporary medical research indicates that the dysregulation of PCD patterns such as apoptosis, necroptosis, pyroptosis, autophagy, and ferroptosis is closely related to the onset and progression of COPD. Clarifying the molecular mechanisms of PCD in COPD may provide novel perspectives for in-depth understanding and prevention of the disease. Traditional Chinese medicine (TCM) is characterized by holistic regulation. In recent years, extensive research has been conducted in the TCM field focusing on modulating apoptosis, necroptosis, pyroptosis, autophagy, and ferroptosis for the treatment of COPD, yielding remarkable achievements. Therefore, this study systematically explored the molecular mechanism of PCD in COPD and reviewed the potential mechanisms and intervention status of TCM targeting PCD in COPD, aiming to provide insights and references for the clinical prevention, treatment and in-depth research of COPD.
5.Comparison of Wild and Cultivated Bupleurum scorzonerifolium Based on Traditional Quality Evaluation
Changsheng YUAN ; Feng ZHOU ; Xingyu LIU ; Yu SHI ; Yihan WANG ; Huaizhu LI ; Yongliang LI ; Shan GUAN ; Huaizhong GAO ; Yanmeng LIU ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):203-214
ObjectiveTo characterize the quality differences among different germplasm and introduced varieties of Bupleurum scorzonerifolium roots(BSR), and explore the underlying molecular mechanisms, providing a basis for high-quality production and quality control. MethodsWild BSR from Yulin(YLW) served as the quality reference, we conducted comparative analysis among YLW, locally domesticated wild germplasm in Yulin(YLC3), Daqing germplasm introduced and cultivated in Yulin(YLDQC3), and locally cultivated germplasm in Daqing(DQC3). A combination of traditional pharmacognostic methods and modern multi-omics analyses was employed, including macroscopic traits(appearance, odor), microscopic features(proportions of cork, phloem, xylem), cell wall component contents(hemicellulose, cellulose, lignin), carbohydrate contents(starch, water-soluble polysaccharides), marker compound contents(ethanol-soluble extracts, total saponins, liposoluble extracts, and saikosaponins A, B2, C, D), metabolomics, and transcriptomics, in order to systematically characterize quality differences and investigate molecular mechanisms among these samples. ResultsMacroscopically, Yulin-produced BSR(YLW, YLC3, YLDQC3) exhibited significantly greater weight, length, and upper and middle diameters than Daqing-produced BSR(DQC3). Odor-wise, YLW and YLC3 had a a fragrance taste, YLDQC3 had a rancid oil odor, and DQC3 had a sweet and fragrant taste. Microscopically, Yulin germplasm(YLW, YLC3) and Daqing germplasm(YLDQC3, DQC3) shared similar structural features, respectively. However, Yulin germplasm showed significantly higher proportions of cork and phloem, as well as stronger xylem vessel staining intensity compared to Daqing germplasm. Regarding various component contents, Yulin germplasm contained significantly higher levels of ethanol-soluble extracts, total saponins, and saikosaponins A, B2, C, D, while Daqing germplasm had significantly higher levels of hemicellulose, starch, and liposoluble extracts. After introduction to Yulin, the Daqing germplasm(YLDQC3) showed increased starch, water-soluble polysaccharides and liposoluble extracts contents, decreased cell wall component content, but no significant difference in other component contents. Metabolomics revealed that saponins and terpenes accumulated significantly in Yulin germplasm, while alcohols and aldehydes accumulated predominantly in Daqing germplasm. Transcriptomics indicated similar gene expression patterns within the same germplasm but specificity between different germplasms. Integrative metabolomic-transcriptomic analysis identified 145 potential key genes associated with the saikosaponin biosynthesis pathway, including one acetyl-coenzyme A(CoA) acetyltransferase gene(ACAT), one 3-hydroxy-3-methylglutaryl-coenzyme A synthase gene(HMGS), two hydroxymethylglutaryl-CoA(HMG-CoA) reductase genes(HMG), one phosphomevalonate kinase gene(PMK), one 1-deoxy-D-xylose-5-phosphate synthase gene(CLA), one hydroxymethylbuten-1-aldol synthase gene(HDR), two farnesyl pyrophosphate synthase genes(FPPS), one squalene synthase gene(SQS), one β-amyrin synthase gene(BAS), 102 cytochrome P450(CYP450) gene family members, and 32 uridine diphosphate-glucuronosyltransferase(UGT) gene family members. ConclusionAmong the three cultivated types, YLC3 most closely resembles YLW in appearance, microscopic features, contents of major bioactive constituents, metabolomic and transcriptomic profiles. Yulin germplasm exhibits superior saponin synthesis capability compared to Daqing germplasm, and Yulin region is more suitable for the growth of B. scorzonerifolium. Based on these findings, it is recommended that artificial cultivation in northern Shaanxi and similar regions utilize the local Yulin germplasm source cultivated for at least three years.
6.Comparison of Wild and Cultivated Alpiniae Oxyphyllae Fructus Based on Traditional Quality Evaluation
Fengfan WANG ; Yajie XIANG ; Jian FENG ; Wencheng HOU ; Wenlan LI ; Yangyang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):235-244
ObjectiveTo compare the differences between wild Alpiniae Oxyphyllae Fructus(WAOF) and cultivated Alpiniae Oxyphyllae Fructus(CAOF) through a traditional quality evaluation system for medicinal materials. MethodsA total of 10 batches of WAOF and 12 batches of CAOF samples were collected from various regions of Hainan province. Relevant analytical methods from the 2020 edition of the Pharmacopoeia of the People's Republic of China were employed to observe the characteristics of WAOF and CAOF, followed by microscopic identification, thin-layer chromatography(TLC) identification, moisture content(toluene method), total ash, acid-insoluble ash, water-soluble and alcohol-soluble extracts(hot dipping method), water-soluble protein, total polysaccharides and total flavonoids(ultraviolet spectrophotometry), and volatile oil content(method A under general rule 2204). The contents of five active components(protocatechuic acid, chrysin, kaempferol, tectochrysin and nootkatone) were quantified using ultra-performance liquid chromatography(UPLC), and the antioxidant activity was evaluated. Building upon traditional quality evaluation of AOF, quantitative measurements were conducted on its appearance traits including diameter, length, plumpness(diameter/length ratio), and color. Canonical correlation analysis was performed using SPSS 26.0 to explore relationships between appearance traits and intrinsic quality. ResultsNo significant differences were observed between WAOF and CAOF in microscopic observation, TLC identification, moisture content, protocatechuic acid content, kaempferol content, odor, or antioxidant activity measured by 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) method. WAOF exhibited significantly higher levels in water-soluble extracts, alcohol-soluble extracts, total polysaccharide content, water-soluble protein content, 100-grain weight, length, and total color difference(ΔE*ab) compared to CAOF(P<0.01). In contrast, CAOF showed significantly higher levels of total ash, acid-insoluble ash, content of total flavonoids, volatile oil content, chrysin content, tectochrysin content, nootkatone content, diameter, plumpness, lightness(L*), red-green chromaticity(a*), yellow-blue chromaticity(b*), and antioxidant activity measured by 1,1-diphenyl-2-picrylhydrazyl(DPPH) method compared to WAOF(P<0.01). Correlation analysis between 7 phenotypic traits and 8 quality traits revealed that among the phenotypic traits, plumpness, L*, a*, and b* exerted significant influence on intrinsic quality. Among the quality traits, total flavonoids, volatile oils, nootkatone, chrysin, and tectochrysin contributed substantially to intrinsic quality. ConclusionPlumpness, L*, a*, and b* of AOF significantly influence its intrinsic quality, and higher values of these parameters indicate relatively superior intrinsic quality. The comprehensive quality evaluation reveals that CAOF samples collected in this study are superior to their wild counterparts.
7.Research progress on the mechanism of traditional Chinese medicine regulating metabolic reprogramming to improve breast cancer
Zhenyu ZHANG ; Weixia CHEN ; Bo FENG ; Jilei LI ; Sizhe WANG ; Meng ZHU ; Chunzheng MA
China Pharmacy 2026;37(2):250-256
Metabolic reprogramming, as one of the core hallmarks of malignant tumors, plays a key role in the occurrence, development and treatment of breast cancer (BC). Abnormal changes in glucose metabolism, amino acid metabolism, lipid metabolism, as well as the tricarboxylic acid (TCA) cycle and oxidative phosphorylation (OXPHOS) pathways significantly influence the pathogenesis and progression of BC. Studies have shown that various active components of traditional Chinese medicine (TCM) (such as berberine, matrine, quercetin, curcumin, etc.) and their compound formulations (e.g. Xihuang pill, Danzhi xiaoyao powder, Yanghe decoction, etc.) can inhibit the proliferation and migration of BC cells and induce apoptosis by regulating key metabolic pathways such as glycolysis, lipid synthesis, and amino acid metabolism. TCM demonstrates multi-target and holistic regulatory advantages in intervening in BC metabolic reprogramming, showing significant potential in modulating key molecules like hypoxia inducible factor-1α, hexokinase-2, pyruvate kinase M2, lactate dehydrogenase A, glucose transporter-1, fatty acid synthase, and signaling pathways such as AKT/mTOR. However, current researches still focus predominantly on glucose metabolism, with insufficient mechanistic studies on lipid metabolism, amino acid metabolism, the TCA cycle, and OXPHOS. The precise targets, molecular mechanisms, and clinical translation value of these interventions require further validation and clarification through more high-quality experimental studies and clinical trials.
8.Role of aspirin in metabolic associated fatty liver disease
Yongqi LI ; Yanqiu LI ; Lina SUN ; Chaoran WANG ; Ying FENG ; Liang WANG ; Xianbo WANG
Journal of Clinical Hepatology 2026;42(1):178-182
Metabolic associated fatty liver disease (MAFLD) is the main type of chronic liver disease in the world, with an increasingly higher incidence rate and a younger age of onset. At present, the treatment of MAFLD mainly depends on lifestyle intervention and comorbidity management, and there is still a lack of effective drugs for MAFLD itself. As a classic nonsteroidal anti-inflammatory drug of the salicylic acid family, aspirin can intervene in the pathological process of MAFLD by regulating lipid metabolism, relieving insulin resistance, reducing liver inflammation and oxidative stress response, exerting an anti-liver fibrosis effect, and inhibiting hepatocellular carcinoma, and therefore, it has the value of preventing disease onset, delaying disease progression, and reversing disease condition. This article systematically reviews the mechanism of action and safety of aspirin in the treatment of MAFLD, in order to provide more drug treatment options for MAFLD patients.
9.Clinical features and molecular mechanism of infantile cholestasis caused by IFT122 gene variants
Jiaqi LI ; Yan LI ; Ruida HE ; Jiayan FENG ; Jianshe WANG
Chinese Journal of Pediatrics 2026;64(1):99-105
Objective:To investigate the clinical characteristics of infantile cholestasis caused by IFT122 gene variants and the molecular mechanism underlying its impact on primary cilia.Methods:The clinical data of an infant with cholestasis from the Children′s Hospital of Fudan University in September 2022 were retrospectively analyzed. The whole-exome sequencing was performed to identify candidate variants, which were validated by Sanger sequencing in the family. Immortalized cell lines were generated using lentiviral infection, followed by immunofluorescence staining to assess the impact of the variants on primary cilia. Intergroup comparisons were performed using the independent sample t-test and Mann-Whitney U test .Results:The proband was a 4-month-old male infant presenting with jaundice, distinctive facial features, and sagittal craniosynostosis. Blood biochemistry indicated elevated direct bilirubin, total bile acids, and transaminases, with markedly increased γ-glutamyltransferase (GGT). Liver pathology demonstrated giant cell hepatitis with cholestasis and bile duct dysplasia. Genetic analysis identified compound heterozygous variants in IFT122 (NM_052989.3) gene c.88G>C (p.Ala30Pro) and c.240G>C (p.Trp80Cys), which co-segregated with the disease in the family. Immunofluorescence analysis demonstrated that the IFT122 gene compound heterozygous missense variants not only significantly reduced the proportion of cilia-positive cells but also led to aberrant ciliary localization of ADP-ribosylation factor-like protein 13B (ARL13B).In addition, ciliary deposition with phosphatidylinositol polyphosphate 5-phosphatase type Ⅳ (INPP5E) was reduced. All differences were statistically significant (all P<0.05). Conclusion:The compound heterozygous missense variants in IFT122 gene not only impair ciliogenesis but also disrupt the ciliary localization of ARL13B and INPP5E, ultimately resulting in high-GGT infantile cholestasis.
10.A Systematic Review of Traditional Chinese Medicine Syndrome Efficacy Evaluation Scales Based on the COSMIN Guidelines
Kailin SU ; Zhenzhen FENG ; Jiajia WANG ; Lu WANG ; Guixiang ZHAO ; Jiansheng LI
Journal of Traditional Chinese Medicine 2026;67(4):416-424
ObjectiveTo systematically evaluate the methodological quality and measurement properties of traditional Chinese medicine (TCM) syndrome efficacy evaluation scales, and to provide evidence-based references for selecting high-quality assessment tools in TCM clinical practice. MethodsChina National Knowledge Infrastructure (CNKI), Wanfang Data, VIP Database, Chinese Biomedical Literature Database (CBM), PubMed, the Cochrane Library, Embase, and Web of Science were searched from inception to April 2, 2025, for studies evaluating the measurement properties of TCM syndrome efficacy evaluation scales. Data were extracted, and the methodological quality and measurement properties of the included scales were assessed according to the consensus-based standards for the selection of health measurement instruments (COSMIN). Recommendation levels were formulated based on the grading of evidence. ResultsA total of 46 studies were included, involving 22 generic syndrome efficacy evaluation scales and 24 disease-specific syndrome efficacy evaluation scales. None of the scales reported cross-cultural validity or measurement error. According to the recommendation grades, 2 scales met Grade A recommendations and are suggested for clinical use; 38 scales were classified as Grade B, indicating potential applicability but requiring further validation; and 6 scales were classified as Grade C, suggesting the need for further refinement. ConclusionExisting TCM syndrome efficacy evaluation scales exhibit substantial variability in methodological quality, incomplete reporting of measurement properties, and insufficient attention to scale revision. Future efforts should emphasize standardized design in the development of TCM syndrome scales, strengthen validation procedures for key measurement properties, and prioritize dynamic revision of scales, thereby providing high-quality tools to support the precise evaluation of syndrome efficacy.

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