1.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
2.Research Tackling Paradigm and Technological Layout Strategies Based on Erectile Dysfunction, A Clinical Dominant Disease of Traditional Chinese Medicine
Qi ZHAO ; Yun CHEN ; Baoxing LIU ; Xuejun SHANG ; Fei SUN ; Xiaozhi ZHAO ; Zhigang WU ; Chao SUN ; Peihai ZHANG ; Wanjun CHENG ; Xing ZHOU ; Zhan QIN ; Yufeng PAN ; Weiwei TAO ; Jianhuai CHEN ; Mei MO ; Xiaoxiao ZHANG ; Xing ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):291-299
To thoroughly implement the strategic deployment outlined in the Opinions of the Central Committee of the Communist Party of China and the State Council on Promoting the Inheritance and Innovative Development of Traditional Chinese Medicine regarding research on dominant diseases of traditional Chinese medicine and to uphold the development philosophy of equal emphasis on traditional Chinese medicine and western medicine,the China Association of Chinese Medicine has fully played a leading academic role by systematically organizing and conducting a series of academic youth salons on clinical dominant diseases of traditional Chinese medicine. On September 13,2024,the 36th Youth Salon on Clinical Dominant Diseases was successfully held in Nanjing,focusing on the advantages of traditional Chinese medicine and the integrative traditional Chinese medicine and western medicine in the diagnosis and treatment of erectile dysfunction (ED). The conference brought together leading experts from traditional Chinese medicine,western medicine,and interdisciplinary fields,facilitating in-depth multidisciplinary discussions that led to key consensus on optimizing traditional Chinese medicine treatment protocols for ED,researching and developing new drugs of traditional Chinese medicine,and advancing interdisciplinary development in traditional Chinese medicine. This salon systematically sorted out the clinical strengths and distinctive features of traditional Chinese medicine in the diagnosis and treatment of ED. Based on current research foundations and clinical needs,it identified key directions for future scientific layout and scientific research tackling: (1) Standardization of syndrome differentiation system of traditional Chinese medicine for ED. (2) Optimization and standardization of intervention methods of integrated traditional Chinese medicine and western medicine. (3) High-quality clinical research guided by evidence-based medicine. (4) In-depth analysis of the pharmacological mechanisms of traditional Chinese medicine in the treatment of ED. (5) Clinical translation and application promotion of new drugs of traditional Chinese medicine. (6) Interdisciplinary integration and innovation in traditional Chinese medicine. For each research direction,key focus areas,expected objectives,and clinical value were further refined,along with the establishment of a scientifically sound priority funding level evaluation system. Therefore,building on the series of salons on the ED-focused dominant diseases of traditional Chinese medicine,this paper provides standardized guidance for clinical practice of traditional Chinese medicine in ED management,effectively contributing to the high-quality development of traditional Chinese medicine. It serves as a valuable reference for national scientific and technological strategic layout, research and development decision-making in new drugs of traditional Chinese medicine,research topic planning,and clinical guideline formulation.
3.Research progress on regulation of antitumor immune function of γδ T cells by active ingredients of traditional Chinese medicine
Binrui WANG ; Zheng CHANG ; Meijing QIN ; Limei ZHAO ; Juanmei MO ; Xiang LU ; Chunhua LU
China Pharmacy 2026;37(13):1773-1777
γδ T cells possess both innate and adaptive immune characteristics. Their antigen recognition is independent of major histocompatibility complex presentation, and they can directly recognize abnormal tumor metabolism, showing unique application potential in tumors with low immune infiltration. This paper systematically sorts out the biological characteristics and antitumor immune functions of γδ T cells, and mainly reviews the relevant mechanisms by which active ingredients of traditional Chinese medicine regulate γδ T cells to exert antitumor immune functions. Glycyrrhiza polysaccharide and Astragalus polysaccharide can enhance the antitumor immune function of γδ T cells by promoting the expression of various cytokines, regulating intestinal flora and alleviating immunosuppressive state. Quercetin and puerarin can strengthen γδ T cell-mediated tumor cell killing by regulating the proliferation of γδ T cells, the expression of cytotoxic effector molecules and related signaling pathways. Toosendanin can potentiate γδ T cell-induced tumor cell killing by down-regulating the expression of anti-apoptotic protein in tumor cells. Artesunate can boost γδ T cell-mediated antitumor immune responses by enhancing the effector function of γδ T cells and weakening tumor immune escape. Future research shall focus on exploring the synergistic effects and molecular mechanisms of the combined application of active ingredients of traditional Chinese medicine and γδ T cell immunotherapy, and identifying the key targets for γδ T cells to exert antitumor immune functions.
4.Research progress on regulation of antitumor immune function of γδ T cells by active ingredients of traditional Chinese medicine
Binrui WANG ; Zheng CHANG ; Meijing QIN ; Limei ZHAO ; Juanmei MO ; Xiang LU ; Chunhua LU
China Pharmacy 2026;37(13):1773-1777
γδ T cells possess both innate and adaptive immune characteristics. Their antigen recognition is independent of major histocompatibility complex presentation, and they can directly recognize abnormal tumor metabolism, showing unique application potential in tumors with low immune infiltration. This paper systematically sorts out the biological characteristics and antitumor immune functions of γδ T cells, and mainly reviews the relevant mechanisms by which active ingredients of traditional Chinese medicine regulate γδ T cells to exert antitumor immune functions. Glycyrrhiza polysaccharide and Astragalus polysaccharide can enhance the antitumor immune function of γδ T cells by promoting the expression of various cytokines, regulating intestinal flora and alleviating immunosuppressive state. Quercetin and puerarin can strengthen γδ T cell-mediated tumor cell killing by regulating the proliferation of γδ T cells, the expression of cytotoxic effector molecules and related signaling pathways. Toosendanin can potentiate γδ T cell-induced tumor cell killing by down-regulating the expression of anti-apoptotic protein in tumor cells. Artesunate can boost γδ T cell-mediated antitumor immune responses by enhancing the effector function of γδ T cells and weakening tumor immune escape. Future research shall focus on exploring the synergistic effects and molecular mechanisms of the combined application of active ingredients of traditional Chinese medicine and γδ T cell immunotherapy, and identifying the key targets for γδ T cells to exert antitumor immune functions.
5.Mechanisms of Core Compound Formula and Compatibility of Traditional Chinese Medicine Components for Prevention and Treatment of Chronic Atrophic Gastritis Based on Huazhuo Jiedu Therapy
Fan GAO ; Xiaoxuan MO ; Fengyue XU ; Jiaye TIAN ; Xinmeng ZHAO ; Jianming JIANG ; Qiuhong GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):51-64
ObjectiveTo reveal the compatibility rules of the core compound formula and traditional Chinese medicine (TCM) components for prevention and treatment of chronic atrophic gastritis (CAG) by Huazhuo Jiedu (turbidity-resolving and detoxifying) therapy through uniform design in view of the potential regulatory pathway of endogenous turbid-toxin retention in CAG, elucidate the potential pathway by which the compound formulas regulate the Wnt/β-catenin-connexin 43 (CX43) signaling axis, demonstrated the efficacy enhancement and toxicity attenuation among the components, thus providing experimental evidence for the scientific application of multi-component, multi-target compatibility in TCM compound formulas. MethodsA mouse model of CAG was established with N-methyl-N-nitrosourea (MNU). Sixty mice were randomly allocated into 5 groups (n=12 per group): blank control, model, low-dose Huazhuo Jiedu core compound formula (HXFF-L, 4.004 g·kg-1), high-dose Huazhuo Jiedu core compound formula (HXFF-H, 8.008 g·kg-1), and positive control (folic acid at 11 mg·kg-1 + vitamin B12 at 14 mg·kg-1). The therapeutic efficacy and molecular mechanism of the core compound formula were evaluated by hematoxylin-eosin (HE) staining, Alcian blue-periodic acid-Schiff (AB-PAS) staining, enzyme-linked immunosorbent assay (ELISA), transmission electron microscopy (TEM), immunofluorescence assay, and Western blot. The chemical components of the core compound formula were detected and effective components were screened by ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Another 120 mice were randomized into blank control, model, and 8 HXFF component-combination groups with different dosages (uniform design groups 1-8, n=12 per group). A uniform design was adopted, with pathological scores of mouse gastric tissue as well as serum levels of pepsinogen Ⅰ (PGⅠ), pepsinogen Ⅱ (PGⅡ), and gastrin 17 (GAS17) set as dependent variables. Multivariate linear regression analysis was performed to predict the optimal proportion of core components in the compound formula and explore its compatibility rules. ResultsPharmacodynamic validation showed that compared with the blank control group, the model group had decreased gastric weight index (P<0.01) and increased pathological damage score and inflammatory factor levels (P<0.01). Compared with the model group, these indicators were reversed in the HXFF-L, HXFF-H, and positive control groups (P<0.05, P<0.01). Immunofluorescence results showed that CX43 expression was downregulated in the model group compared with the blank control group (P<0.01) and upregulated in the HXFF-H and positive control groups compared with the model group (P<0.05, P<0.01). Western blot results showed that compared with the blank control group, the model group demonstrated upregulated protein levels of Wnt and β-catenin in the gastric tissue (P<0.01). Compared with those in the model group, the protein level of Wnt was downregulated in the HXFF-L, HXFF-H, and positive control groups (P<0.05, P<0.01), while that of β-catenin was downregulated only in the HXFF-H group (P<0.05). A total of 25 compounds were identified in the core compound formula, including 9 from Coptidis Rhizoma, 10 from Pogostemonis Herba, 2 from Scorpio, and 4 from Lilii Bulbus. According to the category and origin of compounds, Coptidis Rhizoma alkaloids, Pogostemonis Herba volatile oils, Scorpio peptides, and Lilii Bulbus alkaloids were finally selected for component research. The predicted optimal combination of TCM components was as follows: Coptidis Rhizoma alkaloids at 75 mg·kg-1, Pogostemonis Herba volatile oils at 2.25 mg·kg-1, Scorpio protein extract at 90 mg·kg-1, and Lilii Bulbus alkaloids at 9 mg·kg-1. Coptidis Rhizoma alkaloids, Scorpio protein extract, Pogostemonis Herba volatile oils, and Lilii Bulbus alkaloids can ameliorate pathological lesions of the gastric tissue. Scorpio protein extract exerted positive regulatory effects on the gastric function, while Pogostemonis Herba volatile oils and Lilii Bulbus alkaloids showed negative correlations with partial gastric function indicators. Further investigations are required to clarify their dose-effect relationships and safety profiles. ConclusionThis study, for the first time, predicts the optimal proportion and synergistic regulatory mechanism of the core compound formula of Huazhuo Jiedu therapy, revealing the synergistic compatibility logic of its components. It provides a novel strategy for the precise TCM treatment of CAG and the modern development of compound formulas.
6.Mechanisms of Core Compound Formula and Compatibility of Traditional Chinese Medicine Components for Prevention and Treatment of Chronic Atrophic Gastritis Based on Huazhuo Jiedu Therapy
Fan GAO ; Xiaoxuan MO ; Fengyue XU ; Jiaye TIAN ; Xinmeng ZHAO ; Jianming JIANG ; Qiuhong GUO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):51-64
ObjectiveTo reveal the compatibility rules of the core compound formula and traditional Chinese medicine (TCM) components for prevention and treatment of chronic atrophic gastritis (CAG) by Huazhuo Jiedu (turbidity-resolving and detoxifying) therapy through uniform design in view of the potential regulatory pathway of endogenous turbid-toxin retention in CAG, elucidate the potential pathway by which the compound formulas regulate the Wnt/β-catenin-connexin 43 (CX43) signaling axis, demonstrated the efficacy enhancement and toxicity attenuation among the components, thus providing experimental evidence for the scientific application of multi-component, multi-target compatibility in TCM compound formulas. MethodsA mouse model of CAG was established with N-methyl-N-nitrosourea (MNU). Sixty mice were randomly allocated into 5 groups (n=12 per group): blank control, model, low-dose Huazhuo Jiedu core compound formula (HXFF-L, 4.004 g·kg-1), high-dose Huazhuo Jiedu core compound formula (HXFF-H, 8.008 g·kg-1), and positive control (folic acid at 11 mg·kg-1 + vitamin B12 at 14 mg·kg-1). The therapeutic efficacy and molecular mechanism of the core compound formula were evaluated by hematoxylin-eosin (HE) staining, Alcian blue-periodic acid-Schiff (AB-PAS) staining, enzyme-linked immunosorbent assay (ELISA), transmission electron microscopy (TEM), immunofluorescence assay, and Western blot. The chemical components of the core compound formula were detected and effective components were screened by ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Another 120 mice were randomized into blank control, model, and 8 HXFF component-combination groups with different dosages (uniform design groups 1-8, n=12 per group). A uniform design was adopted, with pathological scores of mouse gastric tissue as well as serum levels of pepsinogen Ⅰ (PGⅠ), pepsinogen Ⅱ (PGⅡ), and gastrin 17 (GAS17) set as dependent variables. Multivariate linear regression analysis was performed to predict the optimal proportion of core components in the compound formula and explore its compatibility rules. ResultsPharmacodynamic validation showed that compared with the blank control group, the model group had decreased gastric weight index (P<0.01) and increased pathological damage score and inflammatory factor levels (P<0.01). Compared with the model group, these indicators were reversed in the HXFF-L, HXFF-H, and positive control groups (P<0.05, P<0.01). Immunofluorescence results showed that CX43 expression was downregulated in the model group compared with the blank control group (P<0.01) and upregulated in the HXFF-H and positive control groups compared with the model group (P<0.05, P<0.01). Western blot results showed that compared with the blank control group, the model group demonstrated upregulated protein levels of Wnt and β-catenin in the gastric tissue (P<0.01). Compared with those in the model group, the protein level of Wnt was downregulated in the HXFF-L, HXFF-H, and positive control groups (P<0.05, P<0.01), while that of β-catenin was downregulated only in the HXFF-H group (P<0.05). A total of 25 compounds were identified in the core compound formula, including 9 from Coptidis Rhizoma, 10 from Pogostemonis Herba, 2 from Scorpio, and 4 from Lilii Bulbus. According to the category and origin of compounds, Coptidis Rhizoma alkaloids, Pogostemonis Herba volatile oils, Scorpio peptides, and Lilii Bulbus alkaloids were finally selected for component research. The predicted optimal combination of TCM components was as follows: Coptidis Rhizoma alkaloids at 75 mg·kg-1, Pogostemonis Herba volatile oils at 2.25 mg·kg-1, Scorpio protein extract at 90 mg·kg-1, and Lilii Bulbus alkaloids at 9 mg·kg-1. Coptidis Rhizoma alkaloids, Scorpio protein extract, Pogostemonis Herba volatile oils, and Lilii Bulbus alkaloids can ameliorate pathological lesions of the gastric tissue. Scorpio protein extract exerted positive regulatory effects on the gastric function, while Pogostemonis Herba volatile oils and Lilii Bulbus alkaloids showed negative correlations with partial gastric function indicators. Further investigations are required to clarify their dose-effect relationships and safety profiles. ConclusionThis study, for the first time, predicts the optimal proportion and synergistic regulatory mechanism of the core compound formula of Huazhuo Jiedu therapy, revealing the synergistic compatibility logic of its components. It provides a novel strategy for the precise TCM treatment of CAG and the modern development of compound formulas.
7.Sensitive Scalp Alopecia: From Neuroimmune Inflammation to Hair Follicle Miniaturization
Feng-Nian ZHAO ; Yu-Ran WANG ; Hayat NADIF ; Si-Ying MO ; Chun-Xia ZHAO ; Feng-Lin ZHUO ; Min WANG
Progress in Biochemistry and Biophysics 2026;53(10):2753-2766
The sensitive scalp is a complex syndrome characterised by abnormal nerve sensations, such as stinging, itching, burning, and tightness, and diminished tolerance to physical or chemical stimuli that would not typically cause such unpleasant responses, with its pathology involving physical barrier destruction at the stratum corneum level, neural hyperreactivity and sensory nerve sensitization, and local immunological dysregulation and chronic low‑grade inflammation. The homeostasis of the scalp microenvironment plays a crucial regulatory role in maintaining the hair follicle growth cycle, yet this “soil” factor has long been underappreciated in hair loss research, which has traditionally centered on intrinsic follicular defects, such as genetic predisposition or androgen-driven mechanisms. In this review, we summarized the potential mechanisms underlying the imbalance in the “barrier-neural-immune” network in sensitive scalps. Sensitive scalp is further manifested as impaired skin barrier, including increased transepidermal water loss and abnormal lipid secretion. These alterations are accompanied by increased surface pH, excessive secretion of sebum rich in free fatty acids, reduced microbial diversity, and dysbiosis characterized by a reduction in Cutibacterium and an increase in Staphylococcus colonization. Mechanistically, we also analyzed how neurogenic inflammation, mediated by substances such as substance P (SP) and calcitonin gene-related peptide (CGRP), along with persistent low-grade inflammation, disrupts the microenvironment of the hair follicle stem cell niche through mechanisms involving mechanosensitive ion channels (e.g., Piezo1), oxidative stress, and cytokine cascades. When the barrier is compromised, the weakened stratum corneum may facilitate the penetration of external stimuli, sensitizing cutaneous nerve endings. Those hyperresponsive nerves then fire off neuropeptides like substance P and CGRP. These peptides activate mast cells and other immune cells via the MrgprB2 receptor, prompting degranulation and a wave of pro-inflammatory mediators that kick off neurogenic inflammation. At the same time, oxidative stress and inflammatory cytokines drive a persistent low-grade inflammation that directly damages the hair follicle stem cell niche. Physical compression from shrinking hair shafts activates Piezo1 channels, setting off calcium influx and Caspase‑3‑dependent apoptotic signaling. We primarily considered the interactions between inflammatory signaling and the sensitization of the androgen receptor (AR) pathway, as well as the inhibition of the Wnt/β‑catenin pathway, which may subsequently contribute to hair follicle miniaturization and disruption of the growth cycle. The inflammatory microenvironment appears to boost AR expression in dermal papilla cells, making them more sensitive to dihydrotestosterone (DHT). This heightened sensitivity may promote the expression of transforming growth factor-β1 (TGF-β1), TGF-β2, and Dickkopf-1 (DKK1), which suppress the Wnt/β‑catenin pathway—essential for follicle growth and regeneration—potentially accelerating follicular miniaturization and stalling the growth cycle. Coupled with direct inflammatory apoptosis, oxidative stress, and circadian disruption, these routes may collectively speed up hair follicle stem cell depletion and diffuse hair thinning. These findings point to the need for more mechanistic and translational research to test the proposed pathways and explore therapies that target the microenvironment in sensitive scalp-related hair loss. Ultimately, we hope to provide theoretical support for the intervention of hair loss by restoring the steady state of the scalp microenvironment and combining with new precision delivery technologies such as microneedles.
8.Clinical characteristics and prognostic analysis of elderly patients with acute myeloid leukemia
Gang ZHANG ; Xiaohui GAO ; Shuping MO ; Yan ZHOU ; Mengjie WU ; Haibing WU ; Xiaoyan ZHAO ; Minchao YAN ; Yuan LI ; Hui ZENG
China Modern Doctor 2025;63(22):1-4
Objective To investigate the clinical characteristics and prognostic risk factors in elderly patients with acute myeloid leukemia(AML).Methods A retrospective analysis was conducted on the clinical data of 101 elderly AML patients admitted to Affiliated Hospital of Jiaxing University from January 2022 to December 2024.All patients were treated with azacitidine+venetoclax regimen.The clinical characteristics of patients and the risk factors related to prognosis were explored.Results The median follow-up was 14 months.Among the 101 patients,74 achieved complete remission or complete remission with incomplete hematological recovery.The median overall survival(OS)of patients with aged ≥70 years,white blood cell count>50 × 109/L,TP53 mutation,complex chromosomes,and high-risk European leukemia net(ELN)risk stratification was significantly shortened(P<0.05).Multivariate analysis showed that age(HR=0.125,95%CI:0.023-0.662,P=0.015),white blood cell count(HR=0.145,95%CI:0.032-0.662,P=0.013),and ELN risk stratification(HR=100.397,95%CI:14.395-700.207,P<0.001)were all independent influencing factors for OS in elderly AML patients.Conclusion Age,white blood cell count and ELN risk stratification are all independent influencing factors affecting OS in elderly AML patients.
9.Dioscin inhibits IL-17+γδT cells to exert an anti-rheumatoid arthritis effect
Lin-mei PU ; Hao-hong ZHANG ; Chao-yu CHU ; Yuan-yuan NI ; Zhao WU ; Qing-yan MO ; Hong-yun WANG ; Ying XU ; Chun-ping WAN
Chinese Pharmacological Bulletin 2025;41(11):2082-2088
Aim To explore the mechanism by which dioscin regulates IL-17+γδT cells in the treatment of arthritis.Methods A collagen-induced arthritis(CIA)model was established in DBA/1 mice using bovine type Ⅱ collagen.The mice were randomly divid-ed into the CIA model group,methotrexate(MTX)positive control group,and dioscin low-dose(Dioscin-L),medium-dose(Dioscin-M),and high-dose(Dios-cin-H)groups.After intervention,the therapeutic effects were evaluated using scoring methods.Joint pathological damage was analyzed by hematoxylin and eosin(HE)staining.The levels of anti-collagen-spe-cific antibodies and the pro-inflammatory cytokine IL-17 were measured by ELISA.The expressions of γδT cells and their subtypes,as well as the secretion level of IL-17,were detected by flow cytometry.Results Dioscin significantly reduced the arthritis severity score in collagen-induced arthritis(CIA)mice,alleviated joint pathological damage,inhibited the production of IL-17 by splenic lymphocytes and the levels of anti-col-lagen-specific antibodies total IgG and IgG3,and de-creased the proportion of γδT cells in the lymph nodes,splenic γδT cells,and the Vδ4+T-cell subset.The level of IL-17 produced by the Vδ4 subtype in the lymph nodes of the intervention groups was lower than that in the model group,but the difference was not sta-tistically significant.Conclusion Dioscin has signifi-cant therapeutic effect on CIA,and its mechanism may be through the inhibition of γδT cells,but it is unlikely to be related to IL-17 derived from γδT cells.
10.Phenotypic Function of Legionella pneumophila Type I-F CRISPR-Cas.
Ting MO ; Hong Yu REN ; Xian Xian ZHANG ; Yun Wei LU ; Zhong Qiu TENG ; Xue ZHANG ; Lu Peng DAI ; Ling HOU ; Na ZHAO ; Jia HE ; Tian QIN
Biomedical and Environmental Sciences 2025;38(9):1105-1119
OBJECTIVE:
CRISPR-Cas protects bacteria from exogenous DNA invasion and is associated with bacterial biofilm formation and pathogenicity.
METHODS:
We analyzed the type I-F CRISPR-Cas system of Legionella pneumophila WX48, including Cas1, Cas2-Cas3, Csy1, Csy2, Csy3, and Cas6f, along with downstream CRISPR arrays. We explored the effects of the CRISPR-Cas system on the in vitro growth, biofilm-forming ability, and pathogenicity of L. pneumophila through constructing gene deletion mutants.
RESULTS:
The type I-F CRISPR-Cas system did not affect the in vitro growth of wild-type or mutant strains. The biofilm formation and intracellular proliferation of the mutant strains were weaker than those of the wild type owing to the regulation of type IV pili and Dot/Icm type IV secretion systems. In particular, Cas6f deletion strongly inhibited these processes.
CONCLUSION
The type I-F CRISPR-Cas system may reduce biofilm formation and intracellular proliferation in L. pneumophila.
Legionella pneumophila/pathogenicity*
;
CRISPR-Cas Systems
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Biofilms/growth & development*
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Phenotype
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Bacterial Proteins/metabolism*
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Gene Deletion

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