1.Mechanism of Wenyang jieyu granules regulating NLRP3/ASC/Caspase-1 pathway on antidepressant effect in rats
Shuang MENG ; Jie ZHAO ; Xinxin WANG ; Dandan TAN ; Xiaorong ZHOU ; Huimin SUN ; Xiaojuan MA ; Zhenyu FENG
China Pharmacy 2026;37(11):1440-1446
OBJECTIVE To explore the antidepressant mechanism of Wenyang jieyu granules (WYJYG) via the NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/apoptosis-associated speck-like protein containing a CARD (ASC)/Caspase-1 pathway. METHODS A rat model of depression was established by chronic unpredictable mild stress combined with single-housing for 42 consecutive days.The experiment set up blank group, model group, MCC950 (NLRP3 inflammasome inhibitor) group (10 mg/kg), fluoxetine group (positive control,2.08 mg/kg),low-dose WYJYG(3.78 g/kg) and high-dose WYJYG group (7.56 g/kg),with 10 rats in each group. From the 22nd day of the experiment, rats in the fluoxetine group, low-dose and high-dose WYJYG groups were intragastrically administered with the corresponding drugs and intraperitoneally injected with an equal volume of normal saline. Rats in the MCC950 group were intraperitoneally injected with MCC950 at the corresponding concentration and intragastrically administered with an equal volume of distilled water. Rats in the blank group and model group were given an equal volume of distilled water by gavage and an equal volume of normal saline by intraperitoneal injection. All interventions were performed once a day for 21 consecutive days. Behavioral tests were conducted once a week. After the last administration, the contents of ASC, ionized calcium binding adaptor molecule 1 (Iba1), interleukin-1β (IL-1β), and IL-18 in hippocampal tissues were detected. The protein expressions of NLRP3, cluster of differentiation 68 (CD68), Caspase-1, B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein were determined, and neuronal apoptosis was observed. RESULTS After the last administration, compared with the model group, the open-field activity time was significantly prolonged ( P <0.05), and the latency to feed in a novel environment was significantly shortened ( P <0.05) in rats of the high-dose WYJYG group. In hippocampal tissue, the contents of ASC, Iba1, IL-1β, and IL-18, as well as the protein expression levels of NLRP3, Caspase-1, and CD68, and the positive rate of neuronal apoptosis were all significantly decreased/downregulated ( P <0.05). Bcl-2 protein expression was significantly upregulated ( P <0.05), and the density of neuronal apoptosis-positive cells was significantly reduced ( P <0.05). CONCLUSIONS WYJYG play on antidepressant role by inhibiting the NLRP3/ASC/Caspase-1 pathway, reducing microglia-mediated neuroinflammation, and inhibiting hippocampal neurons apoptosis.
2.Role of myosin heavy chain 9 in gut-vascular barrier damage in septic mice
Ruili DING ; Chuanbing ZHAO ; Jie LUO ; Rong CHEN ; Qingtao MENG
Chinese Journal of Anesthesiology 2025;45(4):463-468
Objective:To evaluate the role of myosin heavy chain 9(MYH9) in gut-vascular barrier damage in septic mice.Methods:Eighty SPF C57BL/6J male mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups( n=20 each) by the random number table method: sham operation group(Sham group), sham operation + MYH9 inhibitor blebbistatin group(Sham+ Ble group), cecal ligation and perforation(CLP) group, and CLP+ blebbistatin group(CLP+ Ble group). A mouse sepsis model was established using CLP in anesthetized animals. Blebbistatin solution 5 mg/kg was intraperitoneally injected at 1 h before CLP in Sham+ Ble and CLP+ Ble groups, while the equal volume of phosphate buffer was given instead in Sham and CLP groups. Fourteen mice were randomly selected from each group to observe the survival at 24 h after CLP. Blood samples were taken by apical puncture at 24 h after surgery in the remaining 6 mice in each group for determination of the plasma concentrations of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and interleukin-6(IL-6)(using enzyme-linked immunosorbent assay), expression of plasma membrane vesicle-associated protein(PLVAP) in intestinal microvascular endothelial cells(using immunofluorescence), and expression of MYH9, PLVAP, vascular endothelial calreticulin(VE-cadherin) and β-catenin protein and mRNA(by Western bot or fluorescent quantitative real-time polymerase chain reaction) and for examination of the pathological changes of intestinal tissues. Intestinal damage was assessed and scored according to Chiu. Results:Compared with Sham group, the expression of MYH9 protein and mRNA was significantly down-regulated in Sham+ Ble group, and the survival rate was significantly decreased at 24 h after surgery, Chiu′s scores in intestinal tissues were increased, the plasma concentrations of TNF-α, IL-1β and IL-6 were increased, the expression of PLVAP in intestinal microvascular endothelial cells was up-regulated, the expression of MYH9 and PLVAP protein and mRNA in intestinal tissues was up-regulated, and the expression of VE-cadherin and β-catenin protein and mRNA was down-regulated in CLP group( P<0.05). Compared with CLP group, the survival rate was significantly increased at 24 h after surgery, Chiu′s scores in intestinal tissues were decreased, the plasma concentrations of TNF-α, IL-1β and IL-6 were decreased, the expression of PLVAP in intestinal microvascular endothelial cells was down-regulated, the expression of MYH9 and PLVAP protein and mRNA in intestinal tissues was down-regulated, and the expression of VE-cadherin and β-catenin protein and mRNA was up-regulated in CLP+ Ble group( P<0.05). Conclusions:MYH9 is involved in gut-vascular barrier damage in septic mice.
3.Interpretation of the group standard for measurement technique of central venous pressure
Mingxi ZHAO ; Xin GUAN ; Hong SUN ; Hongming MA ; Yueying FENG ; Meng YU ; Dengxiu ZOU ; Hongbo LUO ; Zunzhu LI ; Jie JING ; Xinjuan WU
Chinese Journal of Nursing 2025;60(1):47-51
Central venous pressure(CVP)is a commonly used hemodynamic index in clinical practice,and the accuracy of its measurement results is influenced by the measurement technical specifications.Based on research evidence and clinical practice,this article interprets the main content of the group standard of CVP measurement technology,and proposes suggestions for its use.It aims to guide clinical nursing staff to correctly understand the principles and methods of CVP measurement,and promote the standardization of nursing practice and the accuracy of measurement results.
4.Analysis of the combined application of enteral nutrition and Trocar puncture with a sump drain in the management of Crohn's disease complicated by intra-abdominal abscesses
Meng-Xuan LI ; Wei-Jie LI ; Dan-Dan XU ; Song-Lin CHEN ; Chao-Gang FAN ; Yun ZHAO
Parenteral & Enteral Nutrition 2025;32(3):146-150
Objective:To investigate the efficacy of enteral nutrition combined with trocar puncture with a sump drain in the treatment of Crohn's disease(CD)complicated by intra-abdominal abscess.Methods:A descriptive case series study was conducted.Clinical data of CD patients with intra-abdominal abscess who received enteral nutrition and trocar puncture with a sump drain at the Intestinal Fistula and Intra-abdominal Infection Center of Nanjing BenQ Hospital between June 2018 and December 2022 were retrospectively collected.Laboratory parameters,including white blood cell(WBC)count,C-reactive protein(CRP),and procalcitonin(PCT),were recorded at baseline(pre-puncture)and on days 1,3,and 7 post-puncture.Changes in clinical indicators before and after intervention were compared.Results:All 19 CD patients with intra-abdominal abscess successfully underwent CT-guided trocar puncture with a sump drain,with fistulography confirming the presence of intestinal fistula.No post-procedural complications,including puncture site infection,new-onset intestinal fistula or intra-abdominal hemorrhage,were observed.The median treatment duration was 58.3 days(range:14~300 days).10 patients achieved spontaneous healing while 9 eventually underwent surgical intervention for unresolved fistulas.Post-puncture WBC levels decreased significantly on days 1,3,and 7 compared to baseline(P<0.05).Serum CRP levels increased markedly on day 1 post-puncture(P<0.05)but declined significantly by days 3 and 7(P<0.05).No statistically significant difference was observed in serum procalcitonin levels before and after puncture(P>0.05).Conclusion:The combination of enteral nutrition and trocar puncture with a sump drain is safe,feasible,and effective for managing CD complicated by intra-abdominal abscess.
5.Mechanism of Reactive Oxygen/Nitrogen Species in Liver Ischemia-Reperfusion Injury and Preventive Effect of Chinese Medicine.
Lei GAO ; Yun-Jia LI ; Jia-Min ZHAO ; Yu-Xin LIAO ; Meng-Chen QIN ; Jun-Jie LI ; Hao SHI ; Nai-Kei WONG ; Zhi-Ping LYU ; Jian-Gang SHEN
Chinese journal of integrative medicine 2025;31(5):462-473
Liver ischemia-reperfusion injury (LIRI) is a pathological process involving multiple injury factors and cell types, with different stages. Currently, protective drugs targeting a single condition are limited in efficacy, and interventions on immune cells will also be accompanied by a series of side effects. In the current bottleneck research stage, the multi-target and obvious clinical efficacy of Chinese medicine (CM) is expected to become a breakthrough point in the research and development of new drugs. In this review, we summarize the roles of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in various stages of hepatic ischemia-reperfusion and on various types of cells. Combined with the current research progress in reducing ROS/RNS with CM, new therapies and mechanisms for the treatment of hepatic ischemia-reperfusion are discussed.
Reperfusion Injury/drug therapy*
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Reactive Oxygen Species/metabolism*
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Reactive Nitrogen Species/metabolism*
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Humans
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Liver/drug effects*
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Animals
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Medicine, Chinese Traditional
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Drugs, Chinese Herbal/pharmacology*
6.Zedoarondiol Inhibits Neovascularization in Atherosclerotic Plaques of ApoE-/- Mice by Reducing Platelet Exosomes-Derived MiR-let-7a.
Bei-Li XIE ; Bo-Ce SONG ; Ming-Wang LIU ; Wei WEN ; Yu-Xin YAN ; Meng-Jie GAO ; Lu-Lian JIANG ; Zhi-Die JIN ; Lin YANG ; Jian-Gang LIU ; Da-Zhuo SHI ; Fu-Hai ZHAO
Chinese journal of integrative medicine 2025;31(3):228-239
OBJECTIVE:
To investigate the effect of zedoarondiol on neovascularization of atherosclerotic (AS) plaque by exosomes experiment.
METHODS:
ApoE-/- mice were fed with high-fat diet to establish AS model and treated with high- and low-dose (10, 5 mg/kg daily) of zedoarondiol, respectively. After 14 weeks, the expressions of anti-angiogenic protein thrombospondin 1 (THBS-1) and its receptor CD36 in plaques, as well as platelet activation rate and exosome-derived miR-let-7a were detected. Then, zedoarondiol was used to intervene in platelets in vitro, and miR-let-7a was detected in platelet-derived exosomes (Pexo). Finally, human umbilical vein endothelial cells (HUVECs) were transfected with miR-let-7a mimics and treated with Pexo to observe the effect of miR-let-7a in Pexo on tube formation.
RESULTS:
Animal experiments showed that after treating with zedoarondiol, the neovascularization density in plaques of AS mice was significantly reduced, THBS-1 and CD36 increased, the platelet activation rate was markedly reduced, and the miR-let-7a level in Pexo was reduced (P<0.01). In vitro experiments, the platelet activation rate and miR-let-7a levels in Pexo were significantly reduced after zedoarondiol's intervention. Cell experiments showed that after Pexo's intervention, the tube length increased, and the transfection of miR-let-7a minics further increased the tube length of cells, while reducing the expressions of THBS-1 and CD36.
CONCLUSION
Zedoarondiol has the effect of inhibiting neovascularization within plaque in AS mice, and its mechanism may be potentially related to inhibiting platelet activation and reducing the Pexo-derived miRNA-let-7a level.
Animals
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MicroRNAs/genetics*
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Exosomes/drug effects*
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Plaque, Atherosclerotic/genetics*
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Neovascularization, Pathologic/genetics*
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Human Umbilical Vein Endothelial Cells/metabolism*
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Humans
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Blood Platelets/drug effects*
;
Apolipoproteins E/deficiency*
;
Thrombospondin 1/metabolism*
;
CD36 Antigens/metabolism*
;
Platelet Activation/drug effects*
;
Male
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Mice
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Mice, Inbred C57BL
7.Huachansu injection enhances anti-colorectal cancer efficacy of irinotecan and alleviates its induced intestinal toxicity through upregulating UGT1A1-OATP1B3 expression in vitro and in vivo.
Bo JIANG ; Zhao-Yang MENG ; Yu-Jie HU ; Jun-Jun CHEN ; Ling ZONG ; Ling-Yan XU ; Xiang-Qi ZHANG ; Jing-Xian ZHANG ; Yong-Long HAN
Journal of Integrative Medicine 2025;23(5):576-590
OBJECTIVE:
Huachansu injection (HCSI), a promising anti-cancer Chinese medicine injection, has been reported to have the potential for reducing the toxicity of chemotherapy and improving the quality of life for colorectal cancer (CRC) patients. The objective of this study is to explore the synergistic and detoxifying effects of HCSI when used in combination with irinotecan (CPT-11).
METHODS:
To investigate the effect of HCSI on anti-CRC efficacy and intestinal toxicity of CPT-11, we measured changes in the biological behavior of LoVo cells in vitro, and anti-tumor effects in LoVo cell xenograft nude mice models in vivo. Meanwhile, the effect of HCSI on intestinal toxicity and the uridine diphosphate-glucuronosyltransferase 1A1 (UGT1A1) expression was investigated in the CPT-11-induced colitis mouse model. Subsequently, we measured the effect of HCSI and its 13 constituent bufadienolides on the expression of UGT1A1 and organic anion transporting polypeptides 1B3 (OATP1B3) in HepG2 cells.
RESULTS:
The combination index (CI) results showed that the combination of HCSI and CPT-11 exhibited a synergistic effect (CI < 1), which significantly suppressing the LoVo cell migration, enhancing G2/M and S phase arrest, and inhibiting tumor growth in vivo. Additionally, the damage to intestinal tissues was attenuated by HCSI in CPT-11-induced colitis model, while the increased expression of UGT1A1 in HepG2 cells and in mouse was observed.
CONCLUSION
The co-therapy with HCSI alleviated the intestinal toxicity induced by CPT-11 and exerted an enhanced anti-CRC effect. The detoxifying mechanism may be related to the increased expression of UGT1A1 and OATP1B3 by HCSI and its bufadienolides components. The findings of this study may serve as a theoretical insights and strategies to improve CRC patient outcomes. Please cite this article as: Jiang B, Meng ZY, Hu YJ, Chen JJ, Zong L, Xu LY, Zhang XQ, Zhang JX, Han YL. Huachansu injection enhances anti-colorectal cancer efficacy of irinotecan and alleviates its induced intestinal toxicity through upregulating UGT1A1-OATP1B3 expression in vitro and in vivo. J Integr Med. 2025; 23(5):576-590.
Irinotecan/therapeutic use*
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Animals
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Glucuronosyltransferase/genetics*
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Humans
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Colorectal Neoplasms/metabolism*
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Drugs, Chinese Herbal/therapeutic use*
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Mice, Nude
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Mice
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Up-Regulation/drug effects*
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Male
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Xenograft Model Antitumor Assays
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Mice, Inbred BALB C
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Hep G2 Cells
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Cell Line, Tumor
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Intestines/drug effects*
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Amphibian Venoms
8.Antidepressant mechanism of Xiaoyaosan: A perspective from energy metabolism of the brain and intestine.
Meng-Ting XIAO ; Sen-Yan WANG ; Xiao-Ling WU ; Zi-Yu ZHAO ; Hui-Min WANG ; Hui-Min LIU ; Xue-Mei QIN ; Xiao-Jie LIU
Journal of Integrative Medicine 2025;23(6):706-720
OBJECTIVE:
This study investigated the antidepression mechanisms of Xiaoyaosan (XYS), a classic Chinese prescription, from the perspective of energy metabolism in the brain and intestinal tissues.
METHODS:
Chronic unpredictable mild stress model-a classic depression rat model-was established. Effects of XYS on behaviors and gastrointestinal motility of depressed rats were investigated. Effects of XYS on energetic charge (EC), adenosine triphosphate-related enzymes, and key enzymes of energy metabolism in both hippocampus and jejunum tissues of depressed rats were investigated using high-performance liquid chromatography, biochemical analysis, and real-time quantitative polymerase chain reaction, respectively. Spearman correlation analysis was conducted to construct a correlation network of "behavior-brain energy metabolism-intestinal energy metabolism" of depression.
RESULTS:
XYS significantly reduced the abnormal behaviors that observed in depressed rats and increased the EC and the activity of Na+-K+-adenosine triphosphatase (ATPase) and Ca2+-Mg2+-ATPase in hippocampus and jejunum tissues of depressed rats. XYS restored the key energetic pathways that had been interrupted by depression, including glycolysis, tricarboxylic acid cycle, and oxidative phosphorylation. Furthermore, XYS exhibited antidepressive effects in terms of regulating energy metabolism in tissues of both brain and intestine.
CONCLUSION
XYS significantly corrected the disturbances in EC and energy metabolism-related enzymes of both brain and intestinal tissues, alleviating both core and concomitant symptoms of depression. The current findings underscore the role of energy metabolism in the antidepressive activity of XYS, providing a fresh perspective on depression, and novel research strategies for revealing the mechanism of actions of traditional Chinese medicines on multi-site and multi-symptom diseases. Please cite this article as: Xiao MT, Wang SY, Wu XL, Zhao ZY, Wang HM, Liu HM, Qin XM, Liu XJ. Antidepressant mechanism of Xiaoyaosan: A perspective from energy metabolism of the brain and intestine. J Integr Med. 2025; 23(6):706-720.
Animals
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Energy Metabolism/drug effects*
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Antidepressive Agents/therapeutic use*
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Drugs, Chinese Herbal/therapeutic use*
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Brain/drug effects*
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Male
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Depression/metabolism*
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Rats
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Rats, Sprague-Dawley
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Intestines/drug effects*
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Hippocampus/drug effects*
9.Consensus on informed consent for orthodontic treatment
Yang CAO ; Bing FANG ; Zuolin JIN ; Hong HE ; Yuxing BAI ; Lin WANG ; Haiping LU ; Zhihe ZHAO ; Tianmin XU ; Weiran LI ; Min HU ; Jinlin SONG ; Jun WANG ; Fang JIN ; Ding BAI ; Xianglong HAN ; Yuehua LIU ; Bin YAN ; Jie GUO ; Jiejun SHI ; Yongming LI ; Zhihua LI ; Xiuping WU ; Jiangtian HU ; Linyu XU ; Lin LIU ; Yi LIU ; Yanqin LU ; Wensheng MA ; Shuixue MO ; Liling REN ; Shuxia CUI ; Yongjie FAN ; Jianguang XU ; Lulu XU ; Zhijun ZHENG ; Peijun WANG ; Rui ZOU ; Chufeng LIU ; Lunguo XIA ; Li HU ; Weicai WANG ; Liping WU ; Xiaoxing KOU ; Jiali TAN ; Yuanbo LIU ; Bowen MENG ; Yuantao HAO ; Lili CHEN
Chinese Journal of Stomatology 2025;60(12):1327-1336
This consensus was developed by the Orthodontic Society of the Chinese Stomatological Association to provide a systematic, scientific, and practical guideline for informed consent in orthodontic care. Orthodontic treatment is typically lengthy, highly individualized, and involves multiple factors such as growth and development, occlusal function, and facial esthetics. Rapid technological advances and diverse risk profiles make the traditional reliance on orthodontist experience or institutional templates insufficient to ensure patients′ full understanding and autonomous decision-making. To address this, the expert panel conducted extensive reviews of domestic and international guidelines, analyzed representative dispute cases, and performed multicenter patient-clinician surveys. Using a multi-round Delphi method, the group established a standardized informed consent framework covering the initial consultation, treatment, and retention phases. The consensus emphasizes that informed consent is not only a fundamental legal and ethical requirement but also a key step in building trust, improving patient compliance, and enhancing treatment satisfaction. Orthodontists should clearly and comprehensively explain treatment plans, potential risks, uncertainties, and associated costs, while respecting the autonomy of patients or guardians, and maintain continuous communication and dynamic evaluation throughout the treatment process. The release of this consensus provides unified and authoritative guidance for clinical orthodontics, helping to standardize informed consent, enhance its transparency, safeguard patient rights, reduce medical risks, and promote high-quality, sustainable development of orthodontic practice.
10.Changing resistance profiles of Haemophilus influenzae and Moraxella catarrhalis isolates in hospitals across China:results from the CHINET Antimicrobial Resistance Surveillance Program,2015-2021
Hui FAN ; Chunhong SHAO ; Jia WANG ; Yang YANG ; Fupin HU ; Demei ZHU ; Yunsheng CHEN ; Qing MENG ; Hong ZHANG ; Chun WANG ; Fang DONG ; Wenqi SONG ; Kaizhen WEN ; Yirong ZHANG ; Chuanqing WANG ; Pan FU ; Chao ZHUO ; Danhong SU ; Jiangwei KE ; Shuping ZHOU ; Hua ZHANG ; Fangfang HU ; Mei KANG ; Chao HE ; Hua YU ; Xiangning HUANG ; Yingchun XU ; Xiaojiang ZHANG ; Wenen LIU ; Yanming LI ; Lei ZHU ; Jinhua MENG ; Shifu WANG ; Bin SHAN ; Yan DU ; Wei JIA ; Gang LI ; Jiao FENG ; Ping GONG ; Miao SONG ; Lianhua WEI ; Xin WANG ; Ruizhong WANG ; Hua FANG ; Sufang GUO ; Yanyan WANG ; Dawen GUO ; Jinying ZHAO ; Lixia ZHANG ; Juan MA ; Han SHEN ; Wanqing ZHOU ; Ruyi GUO ; Yan ZHU ; Jinsong WU ; Yuemei LU ; Yuxing NI ; Jingrong SUN ; Xiaobo MA ; Yanqing ZHENG ; Yunsong YU ; Jie LIN ; Ziyong SUN ; Zhongju CHEN ; Zhidong HU ; Jin LI ; Fengbo ZHANG ; Ping JI ; Yunjian HU ; Xiaoman AI ; Jinju DUAN ; Jianbang KANG ; Xuefei HU ; Xuesong XU ; Chao YAN ; Yi LI ; Shanmei WANG ; Hongqin GU ; Yuanhong XU ; Ying HUANG ; Yunzhuo CHU ; Sufei TIAN ; Jihong LI ; Bixia YU ; Cunshan KOU ; Jilu SHEN ; Wenhui HUANG ; Xiuli YANG ; Likang ZHU ; Lin JIANG ; Wen HE ; Chunlei YUE
Chinese Journal of Infection and Chemotherapy 2025;25(1):30-38
Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91%of the total clinical isolates and 4.07%of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89%were isolated from children and 33.11%were isolated from adults.More than 90%of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79%in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06%were isolated from children and 19.94%isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0%.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

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