1.Pathogenesis Evolution and Stage-based Treatment of Gout: An Exploration Based on Theory of ''Endogenous Dampness Leading to Bi Syndrome''
Yingjie ZHANG ; Fan YANG ; Ruifang YANG ; Zhuoming ZHENG ; Siwei PENG ; Yan XIAO ; Peng CHEN ; Youxin SU ; Jiemei GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):74-83
Gout is a crystal-associated arthropathy caused by the deposition of monosodium urate crystals and is closely related to purine metabolic disorders and impaired uric acid excretion. It is clinically characterized by hyperuricemia, recurrent joint swelling and pain, and tophus formation. The disease course is divided into three stages: The hyperuricemia stage, acute attack stage, and chronic gouty arthritis stage. Modern medicine has reached a consensus on its pathology, but traditional Chinese medicine (TCM) lacks a systematic stage-specific understanding of gout pathogenesis and its underlying mechanisms, making it difficult to guide precise syndrome differentiation and treatment. By integrating classical TCM theory, clinical practice, and modern medical understanding, and drawing upon descriptions of Bi syndrome caused by endogenous dampness and turbidity in classical texts such as Huangdi Neijing·Ling Shu and Synopsis of the Golden Chamber, our team proposes the pathogenic concept of gout as ''endogenous dampness leading to Bi syndrome'' and the core pathogenesis of ''spleen deficiency with internal retention of dampness-turbidity''. We systematically elucidate the evolution of pathogenesis across different stages and corresponding therapeutic strategies. This study posits that metabolic byproducts such as urate fall under the category of ''endogenous pathogenic dampness-turbidity''. When genetic or dietary factors lead to metabolic abnormalities, it manifests as ''spleen deficiency with impaired transport and transformation'', resulting in ''internal retention of pathogenic dampness-turbidity''. When damp-turbidity stagnates in the blood vessels, serum uric acid levels rise. When it stagnates in the viscera and limbs, monosodium urate crystals deposit in the joints. Triggered by precipitating factors, this leads to gout attacks—the core pathological process of ''endogenous dampness leading to Bi syndrome''. Based on this theory, the stage-specific pathogenic characteristics of gout are proposed: The hyperuricemia stage is characterized by ''spleen deficiency with impaired transport and transformation, internal retention of pathogenic dampness-turbidity'', the acute attack stage is primarily marked by ''dampness-turbidity and static heat obstructing the limbs and joints'', while the chronic stage is defined by ''spleen deficiency with internal retention of pathogenic dampness-turbidity, intermingled with phlegm-stasis binding''. The treatment principle centers on ''strengthening the spleen and draining dampness'' throughout all stages. During the hyperuricemia stage, treatment focuses on ''strengthening the spleen, draining dampness, and eliminating turbidity''. In the acute attack stage, the treatment should "strengthen the spleen, drain dampness, clear heat, eliminate turbidity, alleviate swelling, and relieve pain''. In the chronic stage, the treatments emphasizes to ''strengthen the spleen, drain dampness, transform turbidity, clear heat, resolve phlegm, and activate blood circulation''. This approach has yielded favorable therapeutic outcomes in clinical practice. This theoretical system clarifies the nature of gout as ''spleen deficiency being the root, dampness-turbidity being the secondary manifestation'' and systematically analyzes its pathogenesis evolution process and characteristics. The constructed stage-based treatment protocol has been validated through clinical and basic research, providing systematic theoretical guidance and a practical framework for the precise TCM management of gout, thereby promoting the modernization of TCM pathogenesis theory related to gout.
2.Analysis of Chronic Gouty Arthritis Animal Models Based on Clinical Characteristics of Traditional Chinese and Western Medicine
Yan XIAO ; Siyuan LIN ; Fan YANG ; Qianglong CHEN ; Xiaohua CHEN ; Meiling WANG ; Zhen ZHANG ; Jiali LUO ; Youxin SU ; Jiemei GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):84-92
ObjectiveBased on the clinical characteristics of chronic gouty arthritis (CGA) in both traditional Chinese and western medicine, this study aims to systematically evaluate the clinical concordance of existing CGA animal models, providing recommendations for establishing animal models that align with the pathological characteristics of CGA and the manifestations of traditional Chinese medicine syndromes. MethodsBy comprehensively retrieving Chinese and international databases such as China National Knowledge Infrastructure, Wanfang, VIP Chinese Science and Technology Periodical Database (VIP), and PubMed, all relevant literature on CGA animal models was collected. Based on the guidelines, the diagnostic criteria of both traditional Chinese and western medicine were summarized and organized. The evaluation indicators for the CGA model were constructed with reference to existing evaluation modes, and the CGA animal models were analyzed to systematically evaluate the clinical concordance of existing models. ResultsThe current methods used to construct CGA animal models mainly include monosodium urate crystal induction, high-protein diet induction (poultry lack urate oxidase), and high-fat diet combined with urate oxidase inhibitors and joint injection. Based on 11 pieces of included literature, the traditional Chinese and western medicine scoring data of each model were extracted, and the average scoring values of all models were ultimately calculated. The results show that the average clinical concordances of existing CGA animal models in both traditional Chinese and western medicine are 43.33% and 64.44%, respectively. Among them, the model with the highest clinical concordance rate is the one with a high-fat diet combined with potassium oxonate to induce hyperuricemia plus joint injection, achieving 83.33% clinical concordance in western medicine and 60% in traditional Chinese medicine. This model aligns well with the pathogenic characteristics and pathological changes of clinical CGA. ConclusionAlthough current CGA animal models can simulate some pathological characteristics of CGA, they struggle to comprehensively reflect the complex pathological processes of CGA and the characteristics of traditional Chinese medicine syndromes. Therefore, in the future, it is necessary to establish the CGA animal models that incorporate the clinical disease and syndrome characteristics of traditional Chinese and western medicine and formulate the uniform model evaluation criteria, providing more precise tools for CGA mechanism research and the development of traditional Chinese medicine.
3.Mechanism of Huazhuo Sanjie Chubi Presciption in Regulating Macrophage Polarization and Improving Low-grade Inflammation in Rats with Chronic Gouty Arthritis
Yuwan LI ; Yingjie ZHANG ; Siyuan LIN ; Xiaohua CHEN ; Qianglong CHEN ; Fan YANG ; Jun LIU ; Bingyan CHEN ; Peng CHEN ; Jiemei GUO ; Youxin SU ; Yan XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):93-104
ObjectiveTo evaluate the therapeutic effect of Huazhuo SanJie Chubi presciption (HSCD) on chronic gouty arthritis (CGA) rats with low-grade inflammation and to explore the underlying mechanism with a focus on macrophage polarization. MethodsThe 41 male 6-week-old SD rats were randomly allocated, using the random number table, to a normal group (n=8) and a model group (n =33). CGA with low-grade inflammation was induced in the model group by daily gavage of potassium oxonate (250 mg·kg-1·d-1) and hypoxanthine (300 mg·kg-1·d-1), combined with intra-articular injection of a monosodium urate (MSU) crystal suspension (50 μL, 25 g·L-¹) into the left ankle twice weekly. After 4 weeks of modeling, 3 rats were randomly selected from each group for model validation. The remaining successfully modeled rats were randomly divided into a model group, an HSCD group (10.35 g·kg-1·d-1, gavage once daily), an M1 polarization agonist group (L-methionine sulfoximine, 300 mg·kg-1, subcutaneous injection every other day), an M1 polarization agonist + HSCD group, an M2 polarization inhibitor group (PD0325901, 10 mg·kg-1·d-1, gavage once daily), and M2 polarization inhibitor + HSCD group. The corresponding drug or drug combination was administered according to group assignment, whereas rats in the normal and model groups received 0.5% carboxymethyl cellulose sodium (CMC-Na) vehicle (10.35 g·kg-1·d-1, gavage once daily). All interventions were continued for four weeks. During the intervention period, except for the normal group, potassium oxonate (250 mg·kg⁻¹) and hypoxanthine (300 mg·kg-1) were co-administered by gavage every other day to maintain the model. At the end of treatment, serum uric acid (SUA), ankle joint diameter and joint swelling index were measured. The levels of high-sensitivity C-reactive protein (hs-CRP), interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), chemokine C-C motif ligand 2 (CCL2), S100 calcium-binding protein A8/A9 (S100A8/A9), interleukin-10 (IL-10) and arginase-1 (Arg-1) in serum and joint fluid were determined by enzyme-linked immunosorbent assay (ELISA). High-frequency ultrasound was used to assess MSU deposition in the ankle joint. Hematoxylin-eosin (HE) staining was performed to evaluate synovial histopathological changes. Quantitative Real-time PCR and immunofluorescence were used to detect the mRNA and protein expression of the M1 macrophage polarization markers inducible nitric oxide synthase (iNOS) and the M2 macrophage polarization marker scavenger receptor cysteine-rich type 1 protein M130 (CD163) in synovial tissue. ResultsCompared with the normal group, the model group showed significantly elevated SUA level and joint swelling index, and increased levels of pro-inflammatory cytokines, CCL2, and S100A8/A9 in both serum and joint fluid (P<0.05), accompanied by MSU deposition and synovial inflammation in the ankle joint. The mRNA and protein expression levels of macrophage polarization M1/M2 markers iNOS and CD163 in synovial tissues were also significantly up-regulated (P<0.05). Compared with model group, rats in HSCD group had significantly lower SUA levels, attenuated joint swelling, reduced serum levels of pro-inflammatory cytokines, and decreased levels of CCL2 and S100A8/A9 in both serum and joint fluid, accompanied with alleviated MSU deposition and synovial inflammation (P<0.05). HSCD markedly downregulated the mRNA and protein expression of M1 marker iNOS (P<0.05), whereas it had no significant effect on the expression of M2 marker CD163. Compared with the M1 polarization agonist group, the M1 polarization agonist + HSCD group showed significantly reduced joint swelling, lower serum levels of pro-inflammatory cytokines, and decreased levels of CCL2 and S100A8/A9 in joint fluid (P<0.05). In addition, synovial inflammatory cell infiltration and angiogenesis were attenuated, and iNOS mRNA and protein expression levels were significantly reduced (P<0.05). Compared with the M2 polarization inhibitor group, the M2 polarization inhibitor + HSCD group exhibited reduced joint swelling, decreased levels of CCL2 and S100A8/A9 in joint fluid and ameliorated synovial inflammation (P<0.05), whereas the levels of anti-inflammatory mediators (IL-10, Arg-1) and CD163 mRNA and protein expression were not significantly increased. ConclusionHSCD alleviates low-grade inflammation in CGA rats, at least in part, by inhibiting macrophage polarization toward the M1 phenotype.
4.Biological Risk Control for Infectious Experiments in Cats in Animal Biosafety Level 2 Laboratory
He ZHAO ; Tao ZHANG ; Yuzhou XIAO ; Li LI ; Xuefang AN ; Fan ZHANG
Laboratory Animal and Comparative Medicine 2026;46(2):242-250
Cats, owing to their physiological and immunological similarities with humans, have become increasingly valuable as model animals in virology research, drug development, and vaccine evaluation. They are irreplaceable in studies of feline immunodeficiency virus, feline coronavirus, and other related pathogens. However, cats are temperamentally sensitive, exhibit strong stress responses, and possess well-developed nervous systems as well as sharp claws and teeth. Consequently, the biosafety risks associated with infectious experiments using cats in animal biosafety level 2 laboratory (ABSL-2) are significantly higher than those encountered with conventional rodents. Drawing on long-term ABSL-2 operational experience, this article systematically reviews the entire workflow of infectious experiments in laboratory cats — from animal selection, pre-entry preparation, reception and quarantine, housing management, to infectious experimental procedures and incident response — identifying and addressing critical risk points at each stage. For strain selection, SPF-grade shorthair cats with defined genetic backgrounds and docile temperaments are recommended; sex and age should be scientifically matched to experimental objectives. During pre-entry preparation, emphasis is placed on dual-credential personnel management, health surveillance, standardized disinfection of environments and cages, feed and water standards, and robust record-keeping. During reception and quarantine, standardized protocols are established for transport control, appearance inspection, isolation quarantine, pathogen exclusion, and positive-reinforcement training. During infectious experimentation, a "three-fixed" husbandry principle is clearly implemented: dedicated caretakers, fixed feeding/cleaning times, and fixed cage positions. Disinfectant selection, autoclaving of waste, and daily veterinary rounds are rigorously enforced. Operational risk control includes detailed measures for graded personal protection, animal anesthesia and restraint, zoned operation within biosafety cabinets, and disposal of experimental waste. Contingency plans are formulated to address animal death, escape, personnel exposure, and spills of infectious materials. This study provides a reproducible and scalable technical pathway and operational standard for conducting infectious experiments in laboratory cats in ABSL-2 laboratories, offering a reference for other facilities undertaking similar work.
5.Enhancement Effect of Porcine Inhibin Polyclonal Antibody on Superovulation in C57BL/6J Mice
He ZHAO ; Tao ZHANG ; Li LI ; Yuzhou XIAO ; Xuefang AN ; Fan ZHANG
Laboratory Animal and Comparative Medicine 2026;46(2):271-278
ObjectiveTo prepare rabbit anti-porcine inhibin polypeptide-keyhole limpet hemocyanin(KLH) conjugated polyclonal antibody and evaluate its effect on superovulation in C57BL/6J mice. MethodsNew Zealand white rabbits were immunized with a synthesized porcine inhibin polypeptide conjugated with KLH to produce anti-inhibin serum (AIS, i.e., inhibin polyclonal antibody). Female C57BL/6J mice received intraperitoneal injections of purified AIS in combination with pregnant mare serum gonadotropin (PMSG), followed by human chorionic gonadotropin (hCG) after 48 hours to induce superovulation. Oocytes obtained from superovulation were collected and counted 15 hours post-hCG administration, and the number of 2-cell embryos was assessed 24 hours after in vitro fertilization. ResultsAIS prepared by immunizing New Zealand White rabbits with KLH-conjugated porcine inhibin polypeptide was subjected to titer determination by indirect ELISA, showing titers reaching 1∶ 512 000. SDS-polyacrylamide gel electrophoresis analysis of ammonium sulfate-purified AIS revealed distinct 50 kDa and 25 kDa bands corresponding to the theoretical molecular weights of IgG antibody heavy and light chains, confirming successful production of porcine inhibin polyclonal antibody. Compared with conventional superovulation methods, AIS diluted 10-fold combined with PMSG significantly increased the number of oocytes obtained from superovulation in mice (P<0.05) by approximately 1.5-fold. ConclusionPorcine inhibin polyclonal antibody, as an improved superovulation reagent, can improve superovulation efficiency in C57BL/6J mice, and shows promising prospects for future applications.
6.Analysing differences in volatile organic compounds between purple-brown and yellow-brown Ziziphi Spinosae Semen based on HS-GC-IMS technology
DUAN Xiao ; KANG Bingtao ; ZHAO Fan ; YAN Yonggang ; ZHANG Gang ; PENG Liang
Drug Standards of China 2026;27(1):0028-0036
Objective: To systematically compare the types and contents of volatile organic compounds (VOCs) in purplish-brown and yellow-brown Ziziphi Spinosae Semen, and to provide a scientific basis for their rapid identification and quality evaluation.
Methods: Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) was employed to determine the VOCs in two colored samples. GC-IMS spectra and fingerprint profiles were established, and chemometric methods including principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) were applied to visualize the differences in volatile components.
Results: A total of 44 VOCs were identified, including 1 aldehyde, 9 esters, 10 ketones, 2 enals, 14 alcohols, 3 acids, 2 furans, 1 disulfide, and 2 hydrocarbons. Alcohols and esters were the main volatile components. The fingerprint profiles showed high consistency within groups but significant differences between groups. Both PCA and PLS-DA effectively distinguished the two sample types. Compounds such as pentanol, methyl butyrate, 2-ethylfuran, 2-hexanol, and 2-methylpropenal were identified as key markers for differentiation.
Conclusion: HS-GC-IMS combined with chemometrics accurately distinguishes the volatile compounds of Ziziphi Spinosae Semen with different colors. This method is rapid, sensitive, and suitable for quality control and rapid authentication of Ziziphi Spinosae Semen.
7.Immunodynamic changes in a mouse model of malignant pleural effusion
Xiao-Lei WEI ; Xu GUO ; Chuang-Xin ZHANG ; Qi WANG ; Xiao-Fan LIU ; Ming-Ming SHAO ; Huan-Zhong SHI ; Kan ZHAI
Laboratory Animal Research 2026;42(1):59-67
Background:
Malignant pleural effusion (MPE), a common complication of advanced cancers, is associated with poor prognosis and reduced quality of life. Although host–tumor interactions are known to drive MPE development, the associated immune dynamics during disease progression remain unclear. Using a Lewis lung carcinoma-induced MPE model in C57BL/6JNidfc mice, we systematically evaluated general parameters and immune cell changes at two-day intervals throughout disease progression.
Results:
The day of Lewis lung carcinoma cell injection into the pleural space was designated as day 0. By day 10 post-injection (p.i.), MPE-bearing mice exhibited ~ 10% body weight loss, marking the experimental endpoint. Pleural tumor mass and pleural effusion volume were minimal up to day 4 p.i. but increased sharply from day 6 onward.CD45⁺ immune cell counts rose over time, and days 6, 8, and 10 p.i. marked key stages of MPE progression. On day 6, B cells, T cells, and natural killer cells, but not macrophages and neutrophils, increased significantly compared to earlier timepoints. By day 8, all immune cell subsets except T cells exceeded day 6 levels, and at day 10, natural killer cell numbers declined while others continued to increase. Besides, the numbers of CD8⁺ T cells, Th1 cells, regulatory T cells, and M2 macrophages progressively increased from day 6 to 10. Based on these data, days 6 and 10 were defined as early and advanced MPE stages, respectively, with distinct immune phenotypes. In advanced MPE, CD8⁺ T cells displayed reduced IFN-γ, TNF-α, Granzyme B, Perforin, FasL, and Ki-67, but upregulated PD-1 and CTLA-4 relative to early stage. Similarly, Th1 cells showed decreased IFN-γ, TNF-α, and IL-2 production along with reduced Ki-67 expression. Advanced-stage M2 macrophages exhibited lower MHC-II levels and impaired phagocytosis, but higher PD-L1 and IL-10 production, while neutrophils showed reduced TNF-α release and phagocytic activity.
Conclusions
Our findings characterize the temporal immune dynamics associated with MPE progression in a mouse model, revealing a transition from an early immunostimulatory state to a late immunosuppressive state. This study enhances our understanding of MPE immunopathogenesis and provides a foundation for developing precise, stagespecific therapeutic strategies.
8.Intelligent handheld ultrasound improving the ability of non-expert general practitioners in carotid examinations for community populations: a prospective and parallel controlled trial
Pei SUN ; Hong HAN ; Yi-Kang SUN ; Xi WANG ; Xiao-Chuan LIU ; Bo-Yang ZHOU ; Li-Fan WANG ; Ya-Qin ZHANG ; Zhi-Gang PAN ; Bei-Jian HUANG ; Hui-Xiong XU ; Chong-Ke ZHAO
Ultrasonography 2025;44(2):112-123
Purpose:
The aim of this study was to investigate the feasibility of an intelligent handheld ultrasound (US) device for assisting non-expert general practitioners (GPs) in detecting carotid plaques (CPs) in community populations.
Methods:
This prospective parallel controlled trial recruited 111 consecutive community residents. All of them underwent examinations by non-expert GPs and specialist doctors using handheld US devices (setting A, setting B, and setting C). The results of setting C with specialist doctors were considered the gold standard. Carotid intima-media thickness (CIMT) and the features of CPs were measured and recorded. The diagnostic performance of GPs in distinguishing CPs was evaluated using a receiver operating characteristic curve. Inter-observer agreement was compared using the intragroup correlation coefficient (ICC). Questionnaires were completed to evaluate clinical benefits.
Results:
Among the 111 community residents, 80, 96, and 112 CPs were detected in settings A, B, and C, respectively. Setting B exhibited better diagnostic performance than setting A for detecting CPs (area under the curve, 0.856 vs. 0.749; P<0.01). Setting B had better consistency with setting C than setting A in CIMT measurement and the assessment of CPs (ICC, 0.731 to 0.923). Moreover, measurements in setting B required less time than the other two settings (44.59 seconds vs. 108.87 seconds vs. 126.13 seconds, both P<0.01).
Conclusion
Using an intelligent handheld US device, GPs can perform CP screening and achieve a diagnostic capability comparable to that of specialist doctors.
9.Bioprinting Fabrication and Biomechanical Functional Evaluation of Micro vascular Networks
Dongrui ZHANG ; Xiao LIU ; Yubo FAN
Journal of Medical Biomechanics 2025;40(5):1360-1366
Constructing functional microvascular networks in vitro represents a pivotal step in the creation of engineered tissues,organ-on-chip models,and organoids,holding profound implications for tissue engineering,regenerative medicine,drug screening,and disease modeling.As a cutting-edge bio-manufacturing approach,bioprinting enables the precise deposition of biomaterials,cells,and bioactive molecules to fabricate intricate microvascular networks that faithfully replicate the geometric architecture and functional properties of native microvasculature.This review summarizes the research progress in bioprinting microvascular networks,with a focus on bioprinting technologies,bioinks,and the biomechanical functional evaluation of microvascular networks.
10.Effects of G-CDOP Regimen on Liver Function in the Treatment of Follicular Lymphoma in the Real World
Xiao ZHANG ; Fan XIA ; Cheng XIE ; Changju QU ; Yiduo DING
Herald of Medicine 2025;44(11):1830-1834
Objective To investigate the occurrence of liver function abnormalities in patients with follicular lymphoma(FL)treated with the G-CDOP regimen in the real world.Methods Using the hospital information system,a retrospective collection of clinical data was conducted on inpatients diagnosed with FL and treated with at least four courses of the G-CDOP regimen in the Department of Hematology,the First Affiliated Hospital of Soochow University from September 2021 to August 2023.The analysis focused on the incidence,severity,and types of liver function abnormalities,as well as the timing of these occurrences in relation to medication use and other relevant clinical characteristics.Results The study encompassed a total of 55 patients with FL,out of which 30 patients encountered liver function abnormalities during the G-CDOP regimen treatment,yielding an incidence rate of 54.5%.There were 23 cases(41.8%)of grade 1,3 cases(5.5%)of grade 2,and 4 cases(7.3%)of grade 3 liver function abnormalities.Among them,12 cases(21.8%)were hepatocellular injury type,while the cholestasis type was observed in 4 cases(7.3%),and the mixed type was found in 14 cases(25.5%).Liver function abnormalities appeared in 21 patients(38.2%)during the first cycle of the G-CDOP regimen treatment,5 patients(9.1%)in the second cycle,and 1 patient(1.8%),2 patients(3.6%),and 1 patient(1.8%)in the third,fourth,and fifth cycles.Conclusion The G-CDOP regimen has the potential to impact liver function indicators in FL patients,but most are mild and reversible.However,there are also cases of severe liver injury that need to attract clinical attention.

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