1.Mechanism of Yangjing Zhongyutang in Regulating SIRT1/PGC-1α Signaling Pathway to Promote Mitochondrial Function and Alleviate Oxidative Stress Damage in Rats with Diminished Ovarian Reserve
Ping ZHANG ; Lijuan YANG ; Shenghui CHEN ; Wenliang YAO ; Yuliang ZHOU ; Ling MA ; Huiying WU ; Yanwen XU ; Ziyan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):46-55
ObjectiveTo observe the effects of Yangjing Zhongyutang (YJZYT) on mitochondrial biogenesis and oxidative stress damage mediated by the silent information regulator 1 (SIRT1)/peroxisome proliferator-activated receptor gamma coactivator-1alpha (PGC-1α) signaling pathway in cyclophosphamide (CTX)-induced rats with diminished ovarian reserve (DOR), and to explore its mechanism in improving ovarian reserve function and follicular development. MethodsForty-two 8-week-old female SD rats with normal estrous cycles were randomly divided into a blank control group (n=7) and a model group (n=35). Rats in the model group received a single intraperitoneal injection of CTX (90 mg·kg-1) to establish the DOR model. After modeling, estrous cycles were monitored for 7 consecutive days, and model success was confirmed based on criteria for estrous cycle disruption. After successful modeling, rats were divided into groups for intervention: estradiol valerate group (0.09 mg·kg-1), and YJZYT high-, medium-, and low-dose groups (19.98, 9.99, 5.00 g·kg-1). The blank control group and model group were given an equal volume of distilled water by gavage. All groups received daily gavage once for 4 consecutive weeks. The general state, body weight, and ovarian wet weight of rats were observed and recorded, and the ovarian organ index was calculated. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), anti-Müllerian hormone (AMH), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px). Hematoxylin-eosin (HE) staining was performed to observe ovarian histomorphological changes and follicular development status. Immunofluorescence was used to detect reactive oxygen species (ROS) expression levels. Colorimetric assays were employed to measure adenosine triphosphate (ATP) and malondialdehyde (MDA) content in ovarian tissues. Quantitative Real-time polymerase chain reaction (Real-time PCR) was used to detect mitochondrial DNA (mtDNA) copy number and the mRNA expression levels of key genes including SIRT1, PGC-1α, nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM). Western blot was performed to detect the protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM. ResultsCompared with the blank group, rats in the model group exhibited disrupted estrous cycles, obviously reduced body weight, and decreased ovarian index (P<0.05). Ovarian histopathology revealed cortical thinning, loose structure, and a significant reduction in both primordial and growing follicles (P<0.01). Serum FSH and LH levels were significantly elevated (P<0.01), while E2 and AMH levels were obviously reduced (P<0.05, P<0.01). ATP content and mtDNA copy number decreased in ovarian tissue (P<0.01), ROS expression increased, MDA levels rose, while SOD and GSH-Px activities obviously decreased (P<0.05, P<0.01), mRNA and protein expression levels of SIRT1, PGC-1α, NRF1, and TFAM were obviously downregulated (P<0.05, P<0.01). After treatment, compared with the model group, body weight and ovarian index obviously recovered in rats administered various doses of YJZYT (P<0.05), serum E2 and AMH levels increased, while FSH and LH levels obviously decreased (P<0.05, P<0.01), ovarian tissue ATP content and mtDNA copy number were up-regulated, ROS and MDA levels decreased, and antioxidant enzymes SOD and GSH-Px activity obviously increased (P<0.05, P<0.01), Gene and protein expression levels related to the SIRT1/PGC-1α /NRF1/TFAM signaling pathway were obviously up-regulated compared to the model group (P<0.05, P<0.01), HE staining revealed that ovarian structure gradually recovered to integrity in all treatment groups, with a obviously increase in the number of primordial and growing follicles (P<0.05, P<0.01). Granulosa cells were neatly arranged, indicating marked improvement in ovarian function. ConclusionYJZYT may improve ovarian function and follicular development in rats with diminished ovarian reserve by activating the SIRT1/PGC-1α signaling pathway, promoting mitochondrial biogenesis, enhancing mitochondrial function, and alleviating oxidative stress damage.
2.Mechanism of Huoxue Rongluo Prescription Regulating Bmal1 Gene to Promote Blood-brain Barrier Repair After Ischemic Stroke
Yuanchen LIAO ; Desheng ZHOU ; Qiang MA ; Lei LUO ; Menghao HE ; Lijuan LIU ; Xiaofeng GAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):40-50
ObjectiveTo explore the mechanism of Huoxue Rongluo prescription (HXRLP) in repairing the blood-brain barrier (BBB) after ischemic stroke (IS). MethodsMale C57BL/6 mice were randomly divided into sham operation (Sham) group, cerebral infarction model (MCAO) group, environmental circadian disruption with cerebral infarction model (ECD-MCAO) group, low-, medium-, and high-dose HXRLP (HXRLP-L, M, and H) groups (8.5, 17, 34 g·kg-1·d-1, respectively), and positive drug butylphthalide (NBP) group (0.23 mL·d-1). In the Sham group, only the exposed blood vessels were isolated without suture insertion. In the other groups, the middle cerebral artery occlusion (MCAO) model of mice was prepared. In the ECD-MCAO group, HXRLP groups, and NBP group, the environmental circadian disruption (ECD) model was prepared. The mice in the Sham group, MCAO group, and ECD-MCAO group were given the same volume of soybean oil by gavage, while those in the other groups were given the corresponding drugs by gavage. Samples were collected after 7 consecutive days of administration. The mNSS score was used to evaluate the repair effect of HXRLP on neurological deficits after IS. Hematoxylin-eosin (HE) staining was used to assess the impact of HXRLP on the pathological damage of brain tissue after IS. 2,3,5-Triphenyltetrazolium chloride (TTC) staining and cerebral blood perfusion status were used to evaluate the repair effect of HXRLP on brain tissue damage after IS. Evans blue staining and transmission electron microscopy were used to evaluate the improvement effect of HXRLP on the permeability injury of BBB after IS. Immunofluorescence (IF) staining was used to observe the expression of von Willebrand Factor (vWF), brain and muscle Arnt-like 1 (Bmal1), and Occludin in brain tissue. Western blot was used to detect the protein expression of Bmal1, Occludin, tight junction protein (Claudin-5), vascular endothelial growth factor (VEGF), and angiopoietins(Ang), and related analysis was conducted. ResultsCompared with the Sham group, the MCAO group exhibited significantly aggravated neurological deficits, cerebral infarction volume, brain pathological damage, and BBB leakage (P0.01) and significantly reduced cerebral blood perfusion (P0.01). The expression of Bmal1, vWF, vascular endothelial growth factor A (VEGFA), and Ang in brain tissue was significantly enhanced (P0.01), while the expression of Occludin and Claudin-5 was significantly weakened (P0.01). Compared with the MCAO group, the ECD-MCAO group showed significantly aggravated neurological deficits, cerebral infarction volume, and BBB leakage (P0.01), obviously worsened brain pathological damage (P0.05), significantly reduced cerebral blood perfusion (P0.01), and significantly decreased expression of Bmal1, vWF, VEGFA, Ang, Occludin, and Claudin-5 in brain tissue (P0.01). Compared with the ECD-MCAO group, the HXRLP groups of all doses presented significantly improved neurological deficits, cerebral infarction volume, brain pathological damage, and BBB leakage (P0.01), significantly increased cerebral blood perfusion (P0.01), and enhanced expression levels of Bmal1, vWF, VEGFA, Ang, Occludin, and Claudin-5 in brain tissue (P0.01). ConclusionHXRLP can regulate the clock protein Bmal1 and promote the expression of VEGFA, Ang, Occludin, and Claudin-5, thereby improving BBB damage after IS.
3.Perioperative electroacupuncture for early gastrointestinal function recovery after colorectal cancer surgery:a pilot study protocol
Jiaojiao WU ; Xuqiang WEI ; Shouquan FENG ; Qunhao GU ; Jing LI ; Zi YE ; Chenchen FENG ; Lijuan PEI ; Yanbin PENG ; Jia ZHOU
Journal of Acupuncture and Tuina Science 2025;23(3):239-249
Objective:A preliminary trial is conducted to explore whether perioperative(preoperative,intraoperative,and postoperative)electroacupuncture is more effective than postoperative electroacupuncture in improving gastrointestinal function for colorectal cancer patients.Methods and analysis:The study proposes a randomized,parallel-controlled,single-center trial involving 30 colorectal cancer patients aged 18-79 requiring elective surgery.Participants are randomly assigned to two groups,with equal allocation,where one group receives perioperative electroacupuncture,and the other group receives postoperative electroacupuncture.The treatment duration spans from preoperative to postoperative 72 h,with a subsequent 28-day follow-up period.The primary outcome is the time of first postoperative defecation.The secondary outcomes include the recovery time of postoperative bowel sounds,time of the first flatus,dietary recovery,postoperative gastrointestinal dysfunction frequency,quality of life scale,postoperative pain degree,time of the first ambulation,length of hospital stay,gastrointestinal hormone indicators,and adverse events.The coagulation function test,liver and renal function,and stool and blood routine serve as security biomarkers.The statistical analysis includes the t-test,rank-sum test,Chi-square test,and analysis of variance.A two-sided significance level is set at 5%.Discussion:This study aims to evaluate the feasibility and preliminary effectiveness of perioperative electroacupuncture for gastrointestinal function recovery following colorectal cancer surgery.The study's strengths include its randomized design,well-defined intervention periods,and multi-dimensional outcome assessment.Nevertheless,limitations,such as the small sample size and single-center setting,may affect external validity.The findings will guide protocol refinement and sample size estimation for future large-scale multi-center randomized controlled trials.
4.Effects and mechanisms of anti-enteritis properties of Yang Shuhua
Kunzhao YANG ; Cheng CHEN ; Yu PAN ; Hao ZHOU ; Yixiang ZHOU ; Lijuan SU ; Zhengke HE ; Liting CAO ; Hongxu DU
Chinese Journal of Veterinary Science 2025;45(4):807-817
Through molecular docking and experimental validation,24 SPF male KM mice were ran-domly divided into four groups:the control group(CON),model group(LPS),high-dose Yang Shuhua group(YSH-H),which was gavaged with drug solution containing 1 g/mL crude Yang Shuhua,and the low-dose Yang Shuhua group(YSH-L),which was gavaged with drug solution containing 0.5 g/mL crude Yang Shuhua for a 17-day experimental period.Mouse body weight was recorded during the experiment,and fecal scoring was done 1 h after intraperitoneal injection of LPS.At the end of the experiment,histopathological changes in the jejunum tissue were examined,and the expression of tight junction protein-related genes was detected.Compared to the CON group,mice in the LPS group showed significant decreases in villus height,villus height/crypt depth(V/C)ratio(P<0.01),and significant increase in crypt depth(P<0.01).The mRNA ex-pression levels of ZO-1 and claudin-1 were significantly decreased(P<0.01).Compared to the LPS group,mice in the YSH-H group showed significant decrease in crypt depth(P<0.05),significant increase in villus height(P<0.05),and significant increase in V/C ratio(P<0.01).The YSH-L group showed a significant increase in V/C ratio(P<0.05),a trend of decreased crypt depth,and an increasing trend of villus height.The mRNA expression of ZO-1 in the YSH-H group showed a significant increase(P<0.05)and claudin-1 showed an increasing trend.Luteolin,quercetin,kaempferol,eriodictyol,and the top 5 potential key targets TP53,IL-1β,TNF,AKT1,and IL-10 exhibited molecular docking scores less than-20.95 kJ/mol,indicating strong activity.GO and KEGG enrichment analysis suggested that Yang Shuhua compounds might act on enteritis through the TNF signaling pathway,IL-17 signaling pathway,and PI3K-Akt signaling pathway.The qPCR results showed an upward trend in the mRNA expression levels of TLR4 and AKT1,significant in-crease in Caspase-1 and ASC(P<0.05),and significantly increased expression of NF-κB and NL-RP3(P<0.01).The mRNA expression level of AKT1,TLR4,NLRP3,ASC,Caspase-1,and NF-κB decreased compared to the LPS group,with significant differences in Caspase-1 and NF-κB(P<0.05).This study suggests that Yang Shuhua exerts anti-enteritis effects through multiple compo-nents and pathways,with blockade of the TLR4-AKT-NF-κB pathway possibly being a key thera-peutic mechanism for treating enteritis.
5.Research progress on the construction of mouse models of iron overload
Lijuan AN ; Jinghui ZHOU ; Yufei QI ; Lirong ZENG ; Zheng HAO
Acta Laboratorium Animalis Scientia Sinica 2025;33(4):593-606
Iron overload refers to the pathological state in which the iron content in the body exceeds physiological requirements,Resultsing in the deposition of iron in the organs.Iron overload mouse models are an important tool for the study of iron metabolism disorders and related diseases.This paper summarizes the commonly used modeling method used in the construction of iron overload mouse models,which mainly comprise two categories.(1)exogenous iron overload mouse models,constructed through supplementation with extraenteral iron(injected iron)or intraintestinal iron(oral iron);(2)spontaneous iron overload models,constructed by screening for specific mouse lines or modifying iron metabolism-related genes.The method involving extraenteral iron supplementation has a short modeling duration and a high success rate,making it suitable for single and composite iron overload models.However,the high iron absorption rate may cause toxic reactions;thus,the optimal dose needs to be determined in advance.By contrast,intraintestinal iron supplementation is simple and safe,but has a low iron absorption rate and a long modeling duration.The spontaneous iron overload model is mainly used for specific genetic research studies,which are complex and involve high costs.The various modeling method offer diverse research pathways,spanning from molecular to systemic levels.This diversity is conducive to gaining an in-depth understanding of the pathological mechanisms underlying iron overload and provides an experimental basis for the development of new treatments.
6.Application and effectiveness evaluation of the"cloud-based PAD class"blended teaching model in on-line clinical clerkship teaching of palliative oncology
Xueqin LI ; Hui DONG ; Lijuan ZHANG ; Quan GONG ; Chunyan ZHOU ; Jun LI ; Jiaxin WANG ; Yunhua WANG
Modern Hospital 2025;25(10):1623-1627
Objective To evaluate the effectiveness of a blended teaching model integrating the HBOPPPS framework(Hybrid Bridge-in,Objectives,Pre-assessment,Participatory learning,Post-assessment,Summary)and the"Cloud-based PAD Class"(Presentation-Assimilation-Discussion)in online clinical clerkship teaching for palliative oncology.Methods A self-con-trolled trial was conducted in October 2023,involving 41 undergraduate students majoring in Food Hygiene and Nutrition(Grade 2021)from Kunming Medical University.Phase 1 implemented conventional online teaching(control group),while Phase 2 adopted the"HBOPPPS+Cloud-based PAD Class"blended approach(experimental group).Teaching effectiveness was assessed via post-class examination scores,classroom participation rates,and satisfaction surveys.Results The experimental group dem-onstrated significantly higher outcomes than the control group:examination scores(86.34±4.19 vs.80.02±3.63,P<0.05),classroom participation(51.2%vs.12.2%,P<0.05),and teaching satisfaction(95.1%vs.80.5%,P<0.05).Conclu-sionThe"HBOPPPS+Cloud-based PAD Class"blended teaching model effectively enhances online clinical clerkship outcomes in palliative oncology,significantly improving students' learning initiative,engagement,and classroom participation.
7.Effect and mechanism of Hypericum japonicum Thunb-Prunella vulgaris on reg-ulating NF-κB-NLRP3 pathway to alleviate liver injury
Kunzhao YANG ; Hao ZHOU ; Tao WANG ; Fugui ZHANG ; Zhanghao FU ; Lijuan SU ; Zhengke HE ; Liting CAO ; Hongxu DU
Chinese Journal of Veterinary Science 2025;45(7):1458-1468
Based on network pharmacology,molecular docking,and experimental validation,this study explored the mechanism by which Hypericum japonicum Thunb-Prunella vulgaris treat liver injury.Mice were randomly divided into four groups:a control group(CON),a model group(CCl4),a high-dose drug group(TXD-H),and a low-dose drug group(TXD-L).A mouse liver in-jury model was established using CCl4 induction.The pathological morphology of liver tissue was observed,and the serum levels of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)were measured.Active ingredients of traditional Chinese medicine and targets related to these medicines and diseases were obtained from databases such as TCMSP,PubChem,Swiss Tar-get Prediction,GeneCards,and DisGeNET.The intersection of these targets was used to identify potential drug targets.A network diagram illustrating the relationships between"drug-active com-ponent-intersection target"was constructed using Cytoscape.Potential targets were analyzed using the STRING database for protein-protein interaction(PPI)analysis and the DAVID database for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis.Molecular docking validation was performed using AutoDock Tools software.Subsequently,key target genes,including those related to the NLRP3 inflammasome and pyroptosis,were detected to validate the molecular docking results.Animal experimental results showed that compared to the CON group,serum AST and ALT activities in the CCl4 group mice were significantly increased(P<0.01),while in the TXD-L group,serum AST and ALT activities were significantly decreased(P<0.05)compared to the CCl4 group,and in the TXD-H group,AST and ALT activities were significantly decreased(P<0.01).Through network pharmacology,135 potential targets were i-dentified,with key components found to be tetramethoxyluteolin,quercetin,kaempferol,luteolin and morin based on degree values,and key targets including TNF,SRC,AKT1,EGFR and ESR1.GO enrichment analysis yielded 304 entries,while KEGG enrichment analysis identified 91 biologi-cal pathways.Molecular docking results demonstrated strong binding between the main compo-nents of Hypericum japonicurn Thunb-Prunella vulgaris and key targets.qPCR results showed that compared to the CON group,the CCl4 group exhibited upregulated relative expression levels of SRC,EGFR,TNF-α,AKT1,and IL-18 mRNA,with significant increases in MyD88,NF-κB,IL-1β,NLRP3,Caspase-1,and ASC mRNA(P<0.05),and significant upregulation of TLR4 and GS-DMD mRNA(P<0.01).Compared to the CCl4 group,the TXD-H group displayed significant downregulation of EGFR,AKT1,TLR4,IL-1β,and GSDMD mRNA(P<0.01),significant decrea-ses in TNF-α,MyD88,NF-κB,NLRP3,and ASC mRNA(P<0.05),while SRC,IL-18,and Caspase-1 mRNA showed a downward trend.In conclusion,Hypericum japonicum Thunb-Prunel-la vulgaris exerts hepatoprotective effects through multiple components and pathways,among which inhibition of the NF-κB-NLRP3 pathway to reduce hepatocyte pyroptosis may be one of the important pathways for its protective effects.
8.Expert consensus on infection prevention and control of Creutzfeldt-Jakob disease in medical institutions
Tianxiang GE ; Yangyang JIA ; Chunhui LI ; Jianrong HUANG ; Xiujuan MENG ; Xiaodong GAO ; Jingping ZHANG ; Fu QIAO ; Lijuan XIONG ; Hui LIANG ; Wei LI ; Haiyan LOU ; Wenjuan WU ; Tianxin XIANG ; Jiansen CHEN ; Biao ZHU ; Kaijin XU ; Zhihui ZHOU ; Hongliu CAI ; Meihong YU ; Yan ZHANG ; Yanwan SHANGGUAN ; Haiting FENG ; Hangping YAO ; Lei GUO ; Tieer GAN ; Weihong ZHANG ; Jimin SUN ; Ye LU ; Qun LU ; Meng CAI ; Jin SHEN ; Yunsong YU ; Anhua WU ; Liu-yi LI ; Tingting QU
Chinese Journal of Infection Control 2025;24(4):437-450
Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.
9.Application and effectiveness evaluation of the"cloud-based PAD class"blended teaching model in on-line clinical clerkship teaching of palliative oncology
Xueqin LI ; Hui DONG ; Lijuan ZHANG ; Quan GONG ; Chunyan ZHOU ; Jun LI ; Jiaxin WANG ; Yunhua WANG
Modern Hospital 2025;25(10):1623-1627
Objective To evaluate the effectiveness of a blended teaching model integrating the HBOPPPS framework(Hybrid Bridge-in,Objectives,Pre-assessment,Participatory learning,Post-assessment,Summary)and the"Cloud-based PAD Class"(Presentation-Assimilation-Discussion)in online clinical clerkship teaching for palliative oncology.Methods A self-con-trolled trial was conducted in October 2023,involving 41 undergraduate students majoring in Food Hygiene and Nutrition(Grade 2021)from Kunming Medical University.Phase 1 implemented conventional online teaching(control group),while Phase 2 adopted the"HBOPPPS+Cloud-based PAD Class"blended approach(experimental group).Teaching effectiveness was assessed via post-class examination scores,classroom participation rates,and satisfaction surveys.Results The experimental group dem-onstrated significantly higher outcomes than the control group:examination scores(86.34±4.19 vs.80.02±3.63,P<0.05),classroom participation(51.2%vs.12.2%,P<0.05),and teaching satisfaction(95.1%vs.80.5%,P<0.05).Conclu-sionThe"HBOPPPS+Cloud-based PAD Class"blended teaching model effectively enhances online clinical clerkship outcomes in palliative oncology,significantly improving students' learning initiative,engagement,and classroom participation.
10.Analysis of common non-bacterial pathogens in hospitalized children with acute respiratory infections: a multicenter study in four regions of Fujian Province in 2023
Lin CAI ; Xiaoman GAO ; Fucun ZHU ; Xiuhua LIU ; Wenlong ZHOU ; Shaohua GE ; Lijuan ZHUANG ; Guanglin ZHANG ; Xiaoping LAI ; Ting LIU
Chinese Journal of Preventive Medicine 2025;59(10):1665-1675
Objective:To analyze the distribution and epidemiological characteristics of common non-bacterial pathogens in hospitalized children with acute respiratory tract infections(ARTI)from a multi-center study covering 4 regions in Fujian Province in 2023.Methods:A retrospective cohort study was conducted using medical record analysis.A total of 22 769 hospitalized children with ARTI were enrolled from January to December 2023 across seven regional pediatric medical centers in Fujian Province (covering four major geographical divisions of Fuzhou, Nanping, Sanming and Longyan; all selected hospitals were regional children′s medical centers).Using single-tube multiplex PCR with fragment analysis on a Sanger sequencing platform, the nucleic acids of 11 common non-bacterial respiratory pathogens were tested in nasopharyngeal swabs collected from 22 769 children. These pathogens included influenza A virus(FluA), influenza B virus(FluB), parainfluenza virus(PIV), respiratory syncytial virus (RSV), adenovirus (ADV), human rhinovirus (HRV), human bocavirus (HBoV), human coronavirus (HCoV), human metapneumovirus(HMPV), Mycoplasma pneumoniae(MP), and Chlamydia (Ch). Count data were described as [ n(%)], and the chi-square test/Fisher′s exact test was used to compare the differences in rates between groups. Epidemiological features, including positive detection rates, pathogen profiles, and correlations with region, sex, age and month, were analyzed. Results:Among 22 769 children with ARTI, pathogens were detected in 16 213 cases (71.21%), including 13 340 single infections (58.59%).The detection rates of single pathogens in descending order were human rhinovirus (HRV, 12.95%), Mycoplasma pneumoniae(MP, 12.27%), respiratory syncytial virus(RSV, 11.12%), influenza A virus (Flu-A, 7.98%), parainfluenza virus(PIV, 4.66%), human metapneumovirus(HMPV, 4.60%), adenovirus(ADV, 2.70%), human bocavirus(HBoV, 0.84%), human coronavirus(HCoV, 0.82%), influenza B virus(Flu-B, 0.47%) and Chlamydia(Ch, 0.18%).Mixed infections occurred in 2 873 cases(12.62%), primarily dual infections(2 679 cases).Regional analysis revealed significant disparities:Luoyuan County Hospital (Fuzhou) exhibited the highest total detection rate(86.59%, 1 414/1 633)and mixed infection rate(23.27%, 380/1 633)(both P<0.001), with notably elevated MP (26.39%, 431/1 633);Jian′ou City Hospital(Nanping) ranked second for Flu-A(14.21%, 409/2 879), RSV(13.20%, 380/2 879) and mixed infections(17.12%, 493/2 879);Lianjiang County Hospital(Fuzhou) showed distinct prevalence of Flu-A(10.68%, 130/1 217), PIV(6.00%, 73/1 217), and HBoV(1.73%, 21/1 217); Yong′an City Hospital (Sanming) reported high MP (26.07%, 238/913) and RSV(12.38%, 113/913);Shaowu City Hospital(Nanping) was dominated by MP (18.60%, 407/2 188) and HRV(13.39%, 293/2 188); Tingzhou Hospital(Longyan) had the highest HRV (17.88%, 407/2 276) and Flu-B (0.75%, 17/2 276); and Fuzhou Children′s Hospital showed elevated ADV(3.38%, 394/11 663) and HCoV(1.08%, 126/11 663). Except for Flu-B(0.47%, 108/22 769; P=0.054) and Ch(0.18%, 40/22769; P=0.900), all pathogens and mixed infections exhibited significant regional variations ( P<0.05).Gender analysis indicated higher detection rates of HRV, RSV, Flu-A, ADV, PIV, HBoV and mixed infections in males, while MP, HMPV, Flu-B, HCoV, and Ch were more prevalent in females, with statistically significant differences for HRV and MP (both P<0.001). Age stratification showed the highest overall detection rate in the 3-<6 years group (75.48%; P<0.001): RSV and Ch peaked in infants (<1 year), HRV, PIV, ADV and HBoV in toddlers (1-<3 years), HMPV, HCoV, and mixed infections in preschool children (3-<6 years), and MP, Flu-A and Flu-B in older children (6-<18 years).Analyzing the prevalent months, the monthly prevalence trends of pathogens in various regions are similar.Seasonal trends demonstrated year-round HRV activity (peaking in spring/autumn), MP prevalence in autumn/winter, RSV surges in spring-summer (April-June) and late summer-autumn (August-October), and Flu-A predominanced in winter-spring. Conclusion:Multiplex PCR with fragment analysis demonstrated high diagnostic efficacy. The top 4 non-bacterial pathogens in Fujian Province′s ARTI-hospitalized children in 2023 were HRV, MP, RSV and Flu-A. Pathogen distribution exhibited significant regional, age and seasonal variations, emphasizing the need for targeted prevention strategies.

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