1.Exploring Mechanism of Chaihu Jia Longgu Mulitang in Depressive-like Rats via AMPK/SIRT1/NF-κB/NLRP3 Signaling Pathway
Guang WANG ; Xinhua SONG ; Jie YANG ; Jinyao XU ; Junhua MEI ; Chao CHEN ; Guohua CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):144-152
ObjectiveTo investigate the effects of Chaihu Jia Longgu Mulitang(CJLM) on depression-like behaviors and neuroinflammation in rats subjected to social isolation combined with chronic unpredictable mild stress(CUMS), and to explore the potential underlying mechanisms. MethodsSixty male SD rats were randomly divided into a normal group, a model group, and low-, medium-, and high-dose CJLM groups(2.89, 5.78, 11.56 g·kg-1), as well as a fluoxetine group(10 mg·kg-1). Except for the normal group, all other groups were subjected to social isolation combined with CUMS for 63 d. During the first 35 d, depression models were established only, and from day 36 onward, modeling and drug administration were conducted simultaneously for a total intervention period of 28 d. Depression-like behaviors were evaluated using the sucrose preference test, open-field test, and forced swimming test. Hematoxylin-eosin(HE) staining was performed to observe hippocampal histomorphology. Immunohistochemistry(IHC) was used to detect the expression levels of ionized calcium-binding adapter molecule 1(Iba1) and gasdermin D(GSDMD) proteins in the hippocampus. Western blot analysis was employed to determine the protein expression levels of adenosine 5′-monophosphate(AMP)-activated protein kinase(AMPK) and phosphorylated(p)-AMPK, silent information regulator 1(SIRT1), nuclear factor-κB(NF-κB) and p-NF-κB, NOD-like receptor protein 3(NLRP3), and Caspase-1 in the hippocampus. Real-time quantitative polymerase chain reaction(Real-time PCR) was used to detect the mRNA expression levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, and IL-1β in the hippocampus. ResultsCompared with the normal group, the model group showed a decreased sucrose preference rate(P<0.01), reduced total movement distance(P<0.01), prolonged immobility time(P<0.01), and decreased central zone residence time(P<0.01) in the open-field test, and increased immobility time in the forced swimming test(P<0.01). Hippocampal neuronal structure was damaged. The contents of Iba1 and GSDMD in the hippocampus were significantly increased(P<0.01). The protein expression levels of p-AMPK and SIRT1 in the hippocampus were significantly decreased(P<0.01), whereas the protein expression levels of p-NF-κB, NLRP3, and Caspase-1 were significantly increased(P<0.01). The mRNA expression levels of IL-1β, IL-6, and TNF-α in the hippocampus were significantly upregulated(P<0.01). Compared with the model group, the low-, medium-, and high-dose CJLM groups and the fluoxetine group all were able to reverse depression-like behavioral changes, as evidenced by increased sucrose preference rate, increased total movement distance with shortened immobility time in the open-field test, prolonged central zone residence time, and reduced immobility time in the forced swimming test(P<0.05, P<0.01). Meanwhile, hippocampal neuronal structural damage was alleviated. In the hippocampus, the expression levels of Iba1 and GSDMD were downregulated, the expression levels of p-AMPK and SIRT1 were upregulated, and the abnormal elevations of p-NF-κB, NLRP3, Caspase-1, as well as IL-1β, IL-6, and TNF-α mRNA were suppressed(P<0.05, P<0.01). ConclusionCJLM can ameliorate depression-like behaviors in rats subjected to social isolation combined with CUMS and attenuate hippocampal neuroinflammation and pyroptosis, suggesting that its effects may be associated with the regulation of AMPK/SIRT1/NF-κB/NLRP3 signaling pathway.
2.Preoperative evaluation of lung function in patients with lung cancer using two-phase dual-energy CT perfusion imaging
Lifang LING ; Yizhen JIA ; Qinmin HAO ; Wenzheng XU ; Zhibo WANG ; Jun WANG ; Liang CHEN ; Mei YUAN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(01):79-86
Objective To explore the application value of dual-phase dual-energy CT (DECT) perfusion imaging in preoperative lung function assessment of lung cancer patients. Methods Data were collected from patients with stageⅠA non-small cell lung cancer who underwent surgical treatment in the Department of Thoracic Surgery, the First Affiliated Hospital of Nanjing Medical University, from November 2022 to June 2024. All patients underwent DECT perfusion imaging and pulmonary function testing (PFT) before surgery. PFT observation indicators included ventilation function indicators such as forced expiratory volume in one second (FEV1), forced vital capacity (FVC), 1-second rate (FEV1/FVC), maximal voluntary ventilation (MVV), and diffusion function indicators such as diffusing capacity for carbon monoxide (DLCO) and DLCO per liter of alveolar volume (DLCO/VA). The software eXamine was used to obtain quantitative parameters of DECT perfusion imaging, including volume parameters and perfusion parameters of both lungs and each lung lobe. The correlation between the volume parameters and perfusion parameters of both lungs and the ventilation and diffusion function indicators of the patients, as well as the differences in quantitative parameters of each lung lobe, was analyzed. Results The end-inspiration lung volume and biphasic volume difference were strongly positively correlated with FEV1 and FVC (r=0.636, r=0.682, r=0.614, r=0.624, P<0.001) and moderately positively correlated with MVV and DLCO (r=0.499, r=0.514, r=0.549, r=0.447, P<0.001); the end-expiration lung volume was weakly negatively correlated with DLCO/VA (r=−0.295, P=0.026); the volume ratio was positively correlated with FEV1, FVC, MVV, and MVV% (r=0.424, r=0.399, r=0.415, r=0.310, P<0.05); the end-inspiration iodine content was weakly positively correlated with DLCO/VA% (rs=0.292, P=0.030); the end-expiration iodine content was weakly positively correlated with FEV1, FVC, MVV, DLCO%, and DLCO/VA (r=0.307, r=0.299, r=0.295, r=0.366, r=0.320, P<0.05) and moderately positively correlated with DLCO (r=0.439, P<0.001); the end-inspiration iodine concentration was negatively correlated with FEV1, FVC, MVV, and MVV% (rs=−0.407, rs=−0.426, rs=−0.352, rs=−0.277, P<0.05); the end-expiratory phase iodine concentration was moderately positively correlated with DLCO/VA (r=0.403, P=0.002); both the iodine concentration difference and the iodine concentration ratio were moderately positively correlated with FEV1, FEV1%, FVC, MVV, MVV% (P<0.05). The lung volume and iodine concentration ratio values were both highest in the left upper lung lobe and lowest in the right middle lung lobe; the differences in lung volume, lung volume ratio, intrapulmonary iodine content, and intrapulmonary iodine concentration were all highest in the lower lobes of both lungs and lowest in the middle lobe of the right lung. Conclusion Dual-phase DECT perfusion imaging can accurately assess overall lung function and quantify regional lung function.
3.Characteristics of recent infection among newly confirmed HIV-1 cases in some areas of Hubei during 2017-2022
Cong LIU ; Xingfu SHEN ; Fanghua MEI ; Meng GUO ; Junqiang XU ; Kun CAI
Journal of Public Health and Preventive Medicine 2026;37(2):132-135
Objective To analyze the newly reported HIV-1 infection in several prefectures of Hubei Province,and analyze its influencing factors. Methods The limiting antigen avidity enzyme immunoassay(LAg-avidity EIA,LAg) was conducted on HIV-1 positive samples confirmed by Western blot of Hubei in 2017-2022. The demographic characteristics of the newly infected samples were analyzed by χ2 test.Logistic regression model was used to analyze influencing factors of new infection rate and predict the factors associated with the HIV-1 recent infection. Results There were 403 new cases of HIV-1 from 2017 to 2022 in several prefectures of Hubei Province, of which 77 were newly infected sorted by LAg,with a new infection rate of 19.11%. The newly confirmed HIV-1 persons of whom aged ≤24 years (40.00% new infection ratio), unmarried (29.41%), college or above (31.37%), and from Voluntary counseling and testing testing(VCT) clinics (40.00%) had a higher proportion of new infections, and the difference was statistically significant. Multivariate Logistic regression analysis showed that age ≤24 years old (aOR=4.346,95%CI: 1.342-14.075) and screening from the VCT clinic (aOR=6.761,95%CI: 1.460-31.319) were more likely to be newly infected. Conclusion The proportion of new HIV infection in several prefectures of Hubei province is relatively low in recent years.Further effective publicity and intervention measures for young students and the construction of VCT clinic should be continuously promoted to achieve early diagnosis and treatment.
4.Nanopackaged Astaxanthin Improves Demyelination in Multiple Sclerosis Model Mice by Scavenging Excessive Endogenous Formaldehyde
Wan-Jia LÜ ; Xin ZENG ; Zhi-Qian TONG ; Yang XING ; Xu YANG ; Mei-Na WU ; Ping MA
Progress in Biochemistry and Biophysics 2026;53(2):442-457
ObjectiveMultiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS); however, its underlying neurological pathogenic mechanisms remain incompletely understood. Endogenous formaldehyde (FA), a metabolic byproduct of methylation-demethylation cycles, has recently been implicated in neurotoxicity, oxidative damage, and cognitive impairment. This study aimed to investigate whether excessive FA contributes to myelin sheath demyelination in mice and to evaluate the protective effects and mechanisms of two FA-elimination strategies: sodium bisulfite (NaHSO3), a classical FA scavenger, and polyethylene glycol-modified astaxanthin nanoparticles (PEG-ATX@NPs), a brain-targeted nano-antioxidant formulation. MethodsA chronic demyelination model was established by feeding female C57BL/6J mice a diet containing 0.2% cuprizone (CPZ) for four weeks, followed by a two-week intervention period. Eighty mice were randomly assigned to four groups: NS (normal saline), CPZ+NS, CPZ+NaHSO3, and CPZ+PEG-ATX@NPs. Behavioral tests, including open-field, Y-maze, and pole-climbing assays, were conducted to assess locomotor activity, motor coordination, and working memory. FA levels in serum, corpus callosum, and spinal cord were measured using an Na-FA fluorescent probe and quantified via in vivo and ex vivo fluorescence imaging. Neuroinflammatory responses were evaluated by measuring TNF-α, IL-1β, and IL-6 levels using ELISA, while oxidative stress was assessed by reactive oxygen species (ROS) fluorescence intensity. Demyelination was examined via Luxol fast blue staining, and microglial activation was analyzed by Iba1 immunofluorescence. Correlation analyses were performed to explore relationships among FA levels, inflammatory cytokines, ROS intensity, and behavioral parameters. ResultsCompared with the NS group, mice in the CPZ+NS group exhibited significant weight loss, impaired motor coordination and memory, and markedly reduced myelin regeneration (P<0.05). FA levels and pro-inflammatory cytokines were significantly elevated in serum, corpus callosum, and spinal cord (P<0.05). FA-associated fluorescence in brain and spinal tissues, as well as ROS intensity across all tissues examined, also increased substantially (P<0.05). CPZ treatment induced pronounced microglial activation and severe demyelination in the corpus callosum (P<0.01). Both NaHSO3 and PEG-ATX@NPs effectively reduced FA accumulation in the brain and spinal cord, attenuated demyelination, suppressed microglial activation, decreased inflammatory cytokine levels, and improved motor and cognitive performance. These results confirm that CPZ induced severe demyelination accompanied by oxidative stress, neuroinflammation, and abnormal FA accumulation. Following intervention with either NaHSO3 or PEG-ATX@NPs, endogenous FA levels in the CNS were substantially reduced. Both treatments alleviated demyelination and significantly decreased the number of activated microglia. Levels of TNF-α, IL-1β, and IL-6 in serum, corpus callosum, and spinal cord were downregulated. Behavioral performance improved significantly, as evidenced by enhanced locomotor activity, better coordination, and improved memory function. These findings indicate that both FA-scavenging agents mitigate CPZ-induced biochemical and behavioral abnormalities. ConclusionThis study demonstrates that excessive endogenous FA is closely associated with cognitive impairment, inflammatory dysregulation, and demyelination in a CPZ-induced chronic demyelination mouse model. Clearing abnormally elevated FA effectively reduces neuroinflammation, suppresses microglial overactivation, decreases oxidative stress, and alleviates demyelination, ultimately improving motor and cognitive outcomes in mice. These results suggest that targeting endogenous FA represents a promising therapeutic strategy for MS and other demyelinating disorders. Further investigations are warranted to explore the long-term safety, dosage optimization, and molecular pathways involved in FA-mediated neurotoxicity.
5.Nanopackaged Astaxanthin Improves Demyelination in Multiple Sclerosis Model Mice by Scavenging Excessive Endogenous Formaldehyde
Wan-Jia LÜ ; Xin ZENG ; Zhi-Qian TONG ; Yang XING ; Xu YANG ; Mei-Na WU ; Ping MA
Progress in Biochemistry and Biophysics 2026;53(2):442-457
ObjectiveMultiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS); however, its underlying neurological pathogenic mechanisms remain incompletely understood. Endogenous formaldehyde (FA), a metabolic byproduct of methylation-demethylation cycles, has recently been implicated in neurotoxicity, oxidative damage, and cognitive impairment. This study aimed to investigate whether excessive FA contributes to myelin sheath demyelination in mice and to evaluate the protective effects and mechanisms of two FA-elimination strategies: sodium bisulfite (NaHSO3), a classical FA scavenger, and polyethylene glycol-modified astaxanthin nanoparticles (PEG-ATX@NPs), a brain-targeted nano-antioxidant formulation. MethodsA chronic demyelination model was established by feeding female C57BL/6J mice a diet containing 0.2% cuprizone (CPZ) for four weeks, followed by a two-week intervention period. Eighty mice were randomly assigned to four groups: NS (normal saline), CPZ+NS, CPZ+NaHSO3, and CPZ+PEG-ATX@NPs. Behavioral tests, including open-field, Y-maze, and pole-climbing assays, were conducted to assess locomotor activity, motor coordination, and working memory. FA levels in serum, corpus callosum, and spinal cord were measured using an Na-FA fluorescent probe and quantified via in vivo and ex vivo fluorescence imaging. Neuroinflammatory responses were evaluated by measuring TNF-α, IL-1β, and IL-6 levels using ELISA, while oxidative stress was assessed by reactive oxygen species (ROS) fluorescence intensity. Demyelination was examined via Luxol fast blue staining, and microglial activation was analyzed by Iba1 immunofluorescence. Correlation analyses were performed to explore relationships among FA levels, inflammatory cytokines, ROS intensity, and behavioral parameters. ResultsCompared with the NS group, mice in the CPZ+NS group exhibited significant weight loss, impaired motor coordination and memory, and markedly reduced myelin regeneration (P<0.05). FA levels and pro-inflammatory cytokines were significantly elevated in serum, corpus callosum, and spinal cord (P<0.05). FA-associated fluorescence in brain and spinal tissues, as well as ROS intensity across all tissues examined, also increased substantially (P<0.05). CPZ treatment induced pronounced microglial activation and severe demyelination in the corpus callosum (P<0.01). Both NaHSO3 and PEG-ATX@NPs effectively reduced FA accumulation in the brain and spinal cord, attenuated demyelination, suppressed microglial activation, decreased inflammatory cytokine levels, and improved motor and cognitive performance. These results confirm that CPZ induced severe demyelination accompanied by oxidative stress, neuroinflammation, and abnormal FA accumulation. Following intervention with either NaHSO3 or PEG-ATX@NPs, endogenous FA levels in the CNS were substantially reduced. Both treatments alleviated demyelination and significantly decreased the number of activated microglia. Levels of TNF-α, IL-1β, and IL-6 in serum, corpus callosum, and spinal cord were downregulated. Behavioral performance improved significantly, as evidenced by enhanced locomotor activity, better coordination, and improved memory function. These findings indicate that both FA-scavenging agents mitigate CPZ-induced biochemical and behavioral abnormalities. ConclusionThis study demonstrates that excessive endogenous FA is closely associated with cognitive impairment, inflammatory dysregulation, and demyelination in a CPZ-induced chronic demyelination mouse model. Clearing abnormally elevated FA effectively reduces neuroinflammation, suppresses microglial overactivation, decreases oxidative stress, and alleviates demyelination, ultimately improving motor and cognitive outcomes in mice. These results suggest that targeting endogenous FA represents a promising therapeutic strategy for MS and other demyelinating disorders. Further investigations are warranted to explore the long-term safety, dosage optimization, and molecular pathways involved in FA-mediated neurotoxicity.
6.New advances in etiological diagnosis, prevention, and treatment of infectious keratopathy
International Eye Science 2026;26(1):39-44
Infectious keratitis(IK)is a major blinding eye disease worldwide. Early diagnosis and treatment are crucial for improving prognosis and reducing the economic burden. This article reviews advances in the diagnosis and treatment of IK, aiming to provide new insights for clinical management. In terms of diagnosis, in addition to conventional methods such as microbial culture and confocal microscopy, molecular diagnostic technologies—including high-throughput sequencing(NGS), CRISPR, and nanotechnology-based systems—have significantly enhanced the sensitivity and specificity of multiplex pathogen detection. These approaches are particularly valuable for identifying mixed infections and rare pathogens. Regarding treatment, in response to the growing challenge of drug resistance, novel drug delivery systems employing nanotechnology and bioactive dressings have markedly improved antibacterial efficacy by enhancing drug penetration and retention. Immunomodulatory therapy and photodynamic therapy effectively control inflammatory responses and improve outcomes. Integrated traditional Chinese and Western medicine, as well as microbiome-based therapies, have demonstrated significant advantages in reducing recurrence rates. Stem cell therapy offers new hope for repairing severe corneal damage, while gene therapy—through gene editing or transduction—strengthens the innate defense mechanisms of the cornea and reduces treatment-related side effects.
7.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.
8.Investigation of tumor-suppressive mechanism of Guiqi Yiyuan Extract combined with cisplatin in Lewis lung cancer mice via TXNIP/NLRP3/Caspase-1/GSDMD pathway
Qiong-qiong GUO ; Wen-jie LI ; Jin-tian LI ; Jian-qing LIANG ; Ping TIAN ; Rong HU ; Xu-chao DONG ; Mei-hao XUE ; Long-xin XU
Chinese Traditional Patent Medicine 2025;47(9):2894-2901
AIM To investigate the tumor-suppressive mechanism of Guiqi Yiyuan Extract combined with cisplatin in Lewis lung cancer mice.METHODS Ten intact C57BL/6J mice were assigned to the blank group.Sixty additional mice were developed into Lewis lung cancer models bearing transplanted tumor and subsequently allocated into the model group,the cisplatin group(5 mg/kg),the high-dose Guiqi Yiyuan Extract group(6.6 g/kg),and the low-dose,medium-dose and high-dose Guiqi Yiyuan Extract combined with cisplatin group(1.6,3.3,6.6 g/kg+5 mg/kg),with 10 mice in each group.Mice in the blank and model groups received saline via daily gavage,while treatment groups were administered Guiqi Yiyuan Extract orally(once daily),and cisplatin injection intraperitoneally(once every other day).After 14 days of drug administration,mice were euthanized for endpoint analysis.The following assessments were conducted:general health status and body weight changes monitored throughout the study period;tumor excision and weighing for inhibition rate calculation;histopathological examination of tumors via hematoxylin-eosin(HE)staining;serum quantification of IL-1 β,IL-18 and HMGB1 by ELISA;ultrastructural analysis of tumor cell death using transmission electron microscopy(TEM);spatial localization of TXNIP and GSDMD-N in tumor sections via immunofluorescence(IF);and Western blot detection of TXNIP,NLRP3,Caspase-1,cleaved Caspase-1,GSDMD,GSDMD-N protein expressions in tumor tissues.RESULTS Compared to the model group,the cisplatin group and all combination therapy groups exhibited significant reduction in tumor weight(P<0.05)and increased tumor suppression rate;enhanced tumor tissue necrosis with characteristic pyroptotic morphology;elevated serum levels of IL-1β,IL-18 and HMGB1(P<0.05);and upregulated expressions of pyroptosis-associated proteins TXNIP,NLRP3,Caspase-1,cleaved Caspase-1,GSDMD and GSDMD-N(P<0.05).The high dose combination group demonstrated optimal therapeutic efficacy(P<0.05).CONCLUSION Guiqi Yiyuan Extract enhances cisplatin sensitivity,demonstrating synergistic anti-tumor effects in Lewis lung carcinoma-bearing mice.This combinatorial therapeutic effect likely involves modulation of the TXNIP/NLRP3/Caspase-1/GSDMD pathway.
9.Clinical effects of Qijing Buzhong Yishen Decoction on patients with diabetic nephropathy due to Spleen-Kidney Qi Deficiency and Blood Stasis Obstructing Collaterals
Hao-yu YUAN ; Zi-cheng YE ; Wen-kai XU ; Sai-mei LI
Chinese Traditional Patent Medicine 2025;47(10):3264-3269
AIM To explore the clinical effects of Qijing Buzhong Yishen Decoction on patients with diabetic nephropathy due to Spleen-Kidney Qi Deficiency and Blood Stasis Obstructing Collaterals.METHODS One hundred and two patients were randomly assigned into control group(51 cases)for 3-month intervention of conventional treatment,and observation group(51 cases)for 3-month intervention of both Qijing Buzhong Yishen Decoction and conventional treatment.The changes in clinical effects,blood glucose indices(fasting blood glucose,2 h postprandial blood glucose,HbA1c,TIR),blood lipid indices(TC,TG,LDL-C),renal function indices(UACR,eGFR,24 h urinary albumin excretion rate,sustained urinary albumin excretion rate),inflammatory factors(IL-6,hs-CRP,TNF-α),TCM syndrome scores and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P<0.05).After the treatment,the two groups displayed decreased fasting blood glucose,2 h postprandial blood glucose,HbA1c,UACR,24 h urinary albumin excretion rate,sustained urinary albumin excretion rate,inflammatory factors,TCM syndrome scores(P<0.05),and increased TIR,eGFR(P<0.05),especially for the observation group(P<0.05).No serious adverse reactions were observable in the two groups.CONCLUSION For the patients with diabetic nephropathy due to Spleen-Kidney Qi Deficiency and Blood Stasis Obstructing Collaterals,Qijing Buzhong Yishen Decoction can safely and effectively regulate the blood sugar levels,and improve renal functions.
10.Epidemiological trends and resistance mechanisms of enterobacterales carrying multiple carbapenemases
Ke WANG ; Qi WANG ; Shuo YANG ; Ruiqi WANG ; Xujun CAI ; Jun HOU ; Hong ZOU ; Yan JIN ; Mei LI ; Hui GUO ; Liang JIN ; Xu YANG ; Lin ZHANG ; Hui WANG
Chinese Journal of Laboratory Medicine 2025;48(9):1158-1164
Objective:To characterize the epidemiology, antimicrobial susceptibility, and molecular mechanisms of carbapenem-resistant Enterobacterales (CRE) carrying multiple carbapenemase genes in China, and to provide evidence for infection control and antibiotic stewardship.Methods:From 2016 to 2023, 115 CRE isolates harboring at least two carbapenemase genes were collected from 41 hospitals in 18 provinces across China. Species identification, antimicrobial susceptibility testing, and whole-genome sequencing were performed. Multilocus sequence typing (MLST) and capsular typing were conducted using Kleborate, plasmid replicon types were identified with PlasmidFinder, and a core genome phylogenetic tree was constructed.Results:The majority of isolates belonged to Klebsiella spp. (80.0%, 92/115), followed by E. cloacae (8.7%, 10/115) and E. coli (6.1%, 7/115). The isolates were mainly from Hebei, Beijing, Shandong, and Hunan (60.9%, 70/115), and sputum was the predominant specimen (43.5%, 50/115). The most common genotype was bla KPC+bla NDM (73.0%, 84/115), primarily in Klebsiella spp. (79.8%, 67/84), followed by bla NDM+bla IMP (15.7%, 18/115). The prevalent plasmid replicon types were IncFII (77.5%, 86/111), IncFIB (68.5%, 76/111), IncR (51.4%, 57/111), and IncX3 (20.7%, 23/111). Notably, 88.6% (31/35) of ST11-KL64 K. pneumoniae strains co-harbored IncFII, IncFIB, and IncR plasmids simultaneously. Between 2016 and 2022, the dominant subtype among Klebsiella spp. isolates was bla KPC-2+bla NDM-1 (56.2%, 36/64). In 2023, the bla KPC-2+bla NDM-13 subtype (29.5%, 19/64) emerged and exhibited clonal transmission (single nucleotide polymorphism 2?74 bp) in Hebei, Beijing, and Jilin. Susceptibility testing showed widespread resistance to β-lactams (90.2%-100%). Aztreonam-avibactam, tigecycline, and colistin retained high activity, with susceptibility rates of 90.16%-98.36%. Conclusions:In China, the majority of clinical Enterobacteriaceae strains that harbor multiple carbapenemases are Klebsiella spp. co-producing KPC and NDM enzymes. Dissemination is driven by both clonal expansion of ST11-KL64 and horizontal transfer of IncFII, IncFIB, and IncR plasmids. The recent emergence and regional clonal spread of the bla KPC-2+bla NDM-13 genotype underscore the urgent need for strengthened surveillance and containment measures.


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