1.Differences in deltamethrin resistance and kdr gene mutation in Culex tritaeniorhynchus population in and outside the Yellow Sea wetland
Xiao-er ZHANG ; Zhi-ming WU ; Ye TIAN ; Qian CUI ; Yu-qian JI ; Huan WANG ; Shu-juan YANG ; Yi-chao ZHAO ; Yu WANG ; Hua-yu YIN ; Yu DING ; Guo-jin YAN ; Min-sen ZHAO ; Shou-gang ZHANG ; Bing-dong SONG ; Hong-na CHEN ; Jian GAO ; Wei-fang YANG ; Yu-fu ZHANG ; Hui LIU ; Hong-liang CHU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):101-107
Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.
2.Research on the Application Value of Dynamic Susceptibility Contrast-Perfusion Weighted Imaging Combined with Amide Proton Transfer Imaging in Brian Glioma Grading
Huan CHEN ; Lu HAO ; KALIBUNUR·MAHEMUTI ; Ming-hui ZHU ; Yu-tong ZHU
Progress in Modern Biomedicine 2025;25(17):2811-2819
Objective:To explored the application value of dynamic susceptibility contrast-perfusion weighted imaging(DSC-PWI)combined with amide proton transfer(APT)imaging in brian glioma grading.Methods:Retrospective analysis of clinical data of 100 patients with brian glioma admitted to The Second Affiliated Hospital of Xinjiang Medical University from January 2022 to September 2024.They were divided into a low-grade group(WHO grades Ⅰ-Ⅱ,n=62)and high-grade group(WHO grades Ⅲ-Ⅳ,n=38)according to the World Health Organization(WHO)grading system for tumors.All patients underwent DSC-PWI and APT imaging preoperative examinations.Apparent diffusion coefficien(ADC),cerebral blood volume(CBV),cerebral blood flow(CBF),mean transit time(MTT),time to peak(TTP),maximum APT value(APTmax),minimum APT value(APTmin),the average APT value of the whole lesion(APTwhole)were recorded.The general datas and ADC,CBV,CBF,MTT,TTP,APTmax,APTmin,APTwhole of two group were compared.The influencing factors of high-grade brian glioma were analysed by Multivariate logistic regression.The diagnostic value of high-grade brian glioma was analysed of DSC-PWI combined with APT imaging by receiver operating characteristic(ROC)curve.Results:There was no statistically significant difference in age,gender,lesion location,basilar artery occlusion,necrosis and growth pattern between the two groups(P>0.05),there was a statistically significant difference in the maximum diameter of lesions ≥2 cm and cystic lesions(P<0.05).The ADC of the high-gradel group was lower than that of the low-grade group,while APTmax,APTmin,APTwhole,CBF,CBV,MTT and TTP were all higher than those of the low-grade group(P<0.05).Elevated APTmax,APTmin,CBV,MTT,and TTP were risk factors for high-grade brian glioma(P<0.05),while elevated ADC was a protective factor(P<0.05).The results of ROC analysis showed that,the area under the curve(AUC)values of APTmax,APTmin,ADC,CBV,MTT,and TTP for diagnostic high-grade brian glioma were 0.830,0.868,0.852,0.843,0.803 and 0.827(all P<0.05).The results of ROC analysis showed that,the AUC values of the logistic model for diagnostic high-grade brian glioma was 0.993.Conclusion:Single detection of DSC-PWI and APT imaging parameters has high diagnostic value in brian glioma grading,and combined detection of APTmax,APTmin,ADC,CBV,MTT and TTP imaging parameters can further improve diagnostic accuracy.
3.Research on the Application Value of Dynamic Susceptibility Contrast-Perfusion Weighted Imaging Combined with Amide Proton Transfer Imaging in Brian Glioma Grading
Huan CHEN ; Lu HAO ; KALIBUNUR·MAHEMUTI ; Ming-hui ZHU ; Yu-tong ZHU
Progress in Modern Biomedicine 2025;25(17):2811-2819
Objective:To explored the application value of dynamic susceptibility contrast-perfusion weighted imaging(DSC-PWI)combined with amide proton transfer(APT)imaging in brian glioma grading.Methods:Retrospective analysis of clinical data of 100 patients with brian glioma admitted to The Second Affiliated Hospital of Xinjiang Medical University from January 2022 to September 2024.They were divided into a low-grade group(WHO grades Ⅰ-Ⅱ,n=62)and high-grade group(WHO grades Ⅲ-Ⅳ,n=38)according to the World Health Organization(WHO)grading system for tumors.All patients underwent DSC-PWI and APT imaging preoperative examinations.Apparent diffusion coefficien(ADC),cerebral blood volume(CBV),cerebral blood flow(CBF),mean transit time(MTT),time to peak(TTP),maximum APT value(APTmax),minimum APT value(APTmin),the average APT value of the whole lesion(APTwhole)were recorded.The general datas and ADC,CBV,CBF,MTT,TTP,APTmax,APTmin,APTwhole of two group were compared.The influencing factors of high-grade brian glioma were analysed by Multivariate logistic regression.The diagnostic value of high-grade brian glioma was analysed of DSC-PWI combined with APT imaging by receiver operating characteristic(ROC)curve.Results:There was no statistically significant difference in age,gender,lesion location,basilar artery occlusion,necrosis and growth pattern between the two groups(P>0.05),there was a statistically significant difference in the maximum diameter of lesions ≥2 cm and cystic lesions(P<0.05).The ADC of the high-gradel group was lower than that of the low-grade group,while APTmax,APTmin,APTwhole,CBF,CBV,MTT and TTP were all higher than those of the low-grade group(P<0.05).Elevated APTmax,APTmin,CBV,MTT,and TTP were risk factors for high-grade brian glioma(P<0.05),while elevated ADC was a protective factor(P<0.05).The results of ROC analysis showed that,the area under the curve(AUC)values of APTmax,APTmin,ADC,CBV,MTT,and TTP for diagnostic high-grade brian glioma were 0.830,0.868,0.852,0.843,0.803 and 0.827(all P<0.05).The results of ROC analysis showed that,the AUC values of the logistic model for diagnostic high-grade brian glioma was 0.993.Conclusion:Single detection of DSC-PWI and APT imaging parameters has high diagnostic value in brian glioma grading,and combined detection of APTmax,APTmin,ADC,CBV,MTT and TTP imaging parameters can further improve diagnostic accuracy.
4.Effect of acupuncture pretreatment on PINK1/Parkin pathway-mediated mitophagy in rats with exercise-induced muscle damage.
Yulin GUO ; Ming GAO ; Huan CHEN ; Hui LI ; Xun TIAN ; Yuan ZHAO ; Gang XU ; Junling WEN ; Shaoxiong LI
Chinese Acupuncture & Moxibustion 2025;45(11):1617-1626
OBJECTIVE:
Based on the PTEN-induced hypothetical kinase 1 (PINK1)/Parkin pathway, the effect of acupuncture pretreatment on the expression of mitochondrial autophagy-related proteins in gastrocnemius muscle tissue of rats with exercise-induced muscle damage (EIMD) was observed, and the underlying mechanism of acupuncture pretreatment for the prevention and treatment of EIMD was explored.
METHODS:
Of 88 SD male rats, aged 6 weeks, 8 rats were randomly selected as a blank group, and the remaining 80 rats were randomized into a model group and an acupuncture pretreatment group, with 40 rats in each group. Either the model group or the acupuncture pretreatment group was subdivided randomly into 5 subgroups with 8 rats in each one according to the time points of sample collection, 0 h, 12 h, 24 h, 48 h and 72 h after modeling. An intermittent downhill running centrifugal exercise was carried out on an animal experimental treadmill to establish the EIMD model in the model group and the acupuncture pretreatment group. The rats in the acupuncture pretreatment group received acupuncture at "Guanyuan" (CV6) and bilateral "Zusanli" (ST36), once a day for 20 min each time, for 7 consecutive days before EIMD model preparation. Transmission electron microscopy was used to observe the ultrastructure of gastrocnemius muscle tissue in each group. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in serum were detected by ELISA. Western blot was used to detect the protein expression of PINK1, Parkin, sequestosome 1 (p62) and microtubule-associated protein light chain 3B (LC3B) in rat gastrocnemius muscle tissue. Real-time PCR was adopted to detect the mRNA expression of PINK1, Parkin, p62 and LC3B in rat gastrocnemius muscle tissue.
RESULTS:
Compared with the blank group, the mitochondria of gastrocnemius muscles showed obvious swelling in the 0 h, 12 h, 24 h, and 48 h model subgroups , autophagosomes were formed in the 12 h and 24 h model subgroups, and the mitochondrial morphology returned to normal gradually in the 72 h model subgroup. The serum MDA contents of rats in 5 model subgroups increased (P<0.01, P<0.05). The contents of SOD and CAT in the subgroups of 0 h, 12 h, 24 h and 48 h decreased (P<0.05, P<0.01). The protein and mRNA expression levels of PINK1, Parkin and LC3B in gastrocnemius muscle tissue of rats in 0 h, 12 h and 24 h subgroups were elevated (P<0.01); and the protein and mRNA expression levels of p62 in the 0 h, 12 h, 24 h and 48 h subgroups were reduced (P<0.01, P<0.05). Compared with the model subgroup at the same time point, the myofibril damage and the degree of mitochondrial swelling were mild in each acupuncture pretreatment subgroup, and the numbers of autophagosomes were fewer. The contents of MDA in the acupuncture pretreatment subgroups decreased at 0 h, 12 h, 24 h, and 48 h (P<0.05, P<0.01). The contents of SOD and CAT in the 12 h acupuncture pretreatment subgroup increased (P<0.05, P<0.01). The protein and mRNA expression levels of PINK1 and Parkin in the 0 h, 12 h, and 24 h acupuncture pretreatment subgroups decreased (P<0.01, P<0.05). The protein and mRNA expression levels of LC3B in the 12 h acupuncture pretreatment subgroup decreased (P<0.01), and that of p62 in the 0 h and 24 h acupuncture pretreatment subgroups increased (P<0.01, P<0.05).
CONCLUSION
The intermittent downhill running centrifugal exercise induces the excessive mitochondrial autophagy. Acupuncture pretreatment may attenuate EIMD, and the underlying mechanism is related to the regulation of PINK1/Parkin signaling pathway expression, reducing oxidative stress damage in skeletal muscle cells, and inhibiting mitochondrial autophagy overactivation.
Animals
;
Ubiquitin-Protein Ligases/genetics*
;
Male
;
Rats
;
Acupuncture Therapy
;
Protein Kinases/genetics*
;
Rats, Sprague-Dawley
;
Mitophagy
;
Humans
;
Muscle, Skeletal/metabolism*
;
Physical Conditioning, Animal
;
Muscular Diseases/physiopathology*
;
Signal Transduction
5.Evidence gap between the systematic reviews and clinical concerns in acupuncture and moxibustion for frozen shoulder.
Zhen LUO ; Weijuan GANG ; Xiaoyi HU ; Huan CHEN ; Lu WANG ; Wencui XIU ; Tianyu MING ; Xianghong JING
Chinese Acupuncture & Moxibustion 2025;45(11):1673-1680
OBJECTIVE:
To reveal the gap between the evidence of systematic reviews (SRs) and clinical concerns by systematically summarizing the evidence on acupuncture and moxibustion for frozen shoulder and investigating the concerns and needs of clinicians in treatment with acupuncture and moxibustion for this disease.
METHODS:
The articles of SR and Meta-analysis on acupuncture and moxibustion for frozen shoulder were searched from CNKI, Wanfang, VIP, SinoMed, PubMed, EMbase and Cochrane Library, starting from the inception of each database up to December 31st, 2022. Two researchers screened the articles and extracted data independently. Using AMSTAR-2, the methodological quality of the included studies was evaluated. Based on systematic reviews and expert discussion, a questionnaire on clinical concerns of acupuncture and moxibustion for frozen shoulder was developed and distributed to clinicians. The discrepancies between the evidence and clinical concerns were compared from 5 dimensions, including population, interventions, control measures, outcome indicators and review time points.
RESULTS:
The evidence gaps existed between SRs and clinical concerns. In the existing studies, the needs of personalized treatment were not fully considered in terms of different syndromes/patterns of frozen shoulder and stages of illness, the outcome indicators were not employed properly, the time for outcome measurement was vague, the control groups were set up outside of standardization, and the methodological quality was lower.
CONCLUSION
It is suggested that future studies should improve the quality of methodology, lay more consideration to different patient groups, optimize outcome indicators and standardize the setting of control groups, so as to better meet the needs of patients and achieve the best match between evidence and clinicians' needs.
Humans
;
Acupuncture Therapy
;
Bursitis/therapy*
;
Evidence Gaps
;
Moxibustion
;
Systematic Reviews as Topic
;
Meta-Analysis as Topic
6.Genetic diversity analysis and DNA fingerprinting of Artemisia argyi germplasm resources based on EST-SSR molecular markers.
Yu-Yang MA ; Chang-Jie CHEN ; Ming-Xing WANG ; Yan FANG ; Yu-Huan MIAO ; Da-Hui LIU
China Journal of Chinese Materia Medica 2025;50(9):2356-2364
This study investigates the genetic diversity and evolutionary relationships of different Artemisia argyi germplasm resources to provide a basis for germplasm identification, variety selection, and resource protection. A total of 192 germplasm resources of A. argyi were studied, and EST-based simple sequence repeat(EST-SSR) primers were designed based on transcriptomic data of A. argyi. Polymerase chain reaction(PCR) amplification was performed on these resources, followed by fluorescence capillary electrophoresis to detect genetic diversity and construct DNA fingerprints. From 197 pairs of primers designed, 28 pairs with polymorphic and clear bands were selected. A total of 278 alleles were detected, with an average of 9.900 0 alleles per primer pair and an average effective number of alleles of 1.407 2. The Shannon's diversity index(I) for the A. argyi germplasm resources ranged from 0.148 1 to 0.418 0, with an average of 0.255 7. The polymorphism information content(PIC) ranged from 0.454 5 to 0.878 0, with an average of 0.766 9, showing high polymorphism. Cluster analysis divided the A. argyi germplasm resources into three major groups: Group Ⅰ contained 136 germplasm samples, Group Ⅱ contained 45, and Group Ⅲ contained 11. Principal component analysis also divided the resources into three groups, which was generally consistent with the clustering results. Mantel test results showed that the genetic variation in A. argyi populations was to some extent influenced by geographic distance, but the effect was minimal. Structure analysis showed that 190 germplasm materials had Q≥ 0.6, indicating that these germplasm materials had a relatively homogeneous genetic origin. Furthermore, 8 core primer pairs were selected from the 28 designed primers, which could distinguish various germplasm types. Using these 8 core primers, DNA fingerprints for the 192 A. argyi germplasm resources were successfully constructed. EST-SSR molecular markers can be used to study the genetic diversity and phylogenetic relationships of A. argyi, providing theoretical support for the identification and molecular-assisted breeding of A. argyi germplasm resources.
Artemisia/classification*
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Microsatellite Repeats
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Genetic Variation
;
Expressed Sequence Tags
;
DNA Fingerprinting
;
Phylogeny
;
Polymorphism, Genetic
;
DNA, Plant/genetics*
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Genetic Markers
7.Association of angiotensin Ⅱ type 1 receptor autoantibody and sperm motility in human semen.
Dao-Qin TU ; Yu-Huan YANG ; Gui-Fang YANG ; Yi-Nan ZHANG ; Yao CHEN ; Xue-Ming LI ; Yu-Hui SHI ; Xiao-Li YANG ; Feng WANG
National Journal of Andrology 2025;31(5):387-394
OBJECTIVE:
To investigate the relationship between angiotensin Ⅱ type 1 receptor autoantibody (AT1-AA) and semen parameters. Methods: The semen samples of 820 male patients who were treated in the Reproductive Medicine Center of Taiyuan Central Hospital from August 2022 to August 2023 were retrospectively analyzed. The levels of AT1-AA and Ang Ⅱ of semen were detected by ELISA, and the function of AT1-AA was detected by cardiomyocyte beating assay in suckling rats. The patients were divided into low group, median group and high group according to the OD values of AT1-AA. The differences in general data and semen parameters between different groups were analyzed. And the correlation between AT1-AA level and semen parameters in semen of all study subjects was analyzed by the method of Spearman analysis. And the relationships between AT1-AA OD value, Ang Ⅱ level and semen parameters in the AT1-AA high value group were analyzed as well.
RESULTS:
AT1-AA was present in semen with good function. There was no significant difference in the general data of patients in different AT1-AA levels (P>0.05). In the comparison of semen parameters among the groups with different levels of AT1-AA, there were differences in sperm concentration, PR concentration, NP%, and ALH among the three groups (P<0.05). And AT1-AA OD value was positively correlated with total sperm count, sperm concentration, PR concentration, and NP%, and negatively correlated with semen volume (P<0.05). In the AT1-AA high value group, the OD value of AT1-AA in semen was negatively correlated with inactive sperm, and positively correlated with total motility ([PR+NP]%), curve rate, mean path rate, and ALH. However, there was no correlation between the level of Ang Ⅱ in semen and semen parameters (P>0.05).
CONCLUSION
The presence of AT1-AA in semen may be associated with the promotion of sperm motility.
Male
;
Humans
;
Autoantibodies
;
Sperm Motility
;
Semen
;
Retrospective Studies
;
Receptor, Angiotensin, Type 1/immunology*
;
Animals
;
Rats
;
Angiotensin II
;
Adult
;
Sperm Count
;
Semen Analysis
;
Receptor, Angiotensin, Type 2/immunology*
8.Protective Effects of Ferrostatin-1 on Liver and Kidney Tissues in Mice with Middle and Late Stages of Diabetes
Huan WANG ; Ming-Xing ZHU ; Zhi-Jing WU ; Wei-Wen CHEN ; Yan-Fang ZHENG ; Ming-Qing HUANG
Chinese Journal of Biochemistry and Molecular Biology 2024;40(6):848-856
Diabetes mellitus is a metabolic disease with high incidence and many complications,among which type 2 diabetes mellitus(T2DM)accounts for a large proportion.Current studies have shown that T2DM is accompanied by damage of liver,kidney,and other organs and its complications seriously en-danger human health.Ferroptosis generates many Reactive Oxygen Species(ROS)through the Fenton reaction,and the accumulation of ROS activates Hypoxia Inducible Factor-1(HIF-1α).As a result,the level of vascular endothelial growth factor(VEGF)is increased.Ferrostatin-1(Fer-1),a ferroptosis in-hibitor,has strong antioxidant capacity.Therefore,based on the hypoxia-inducible factor-1α/vascular endothelial growth factor(HIF-1α/VEGF)signaling pathway,we explored the therapeutic effect of Fer-1 on the liver and kidney tissues of diabetic mice.db/db mice(21~22 weeks old)were used as the model of diabetes mellitus.Ferroptosis inhibitor Fer-1 was used as the intervention drug.db/m mice served as the blank control group,and body weight and blood glucose were measured for 4 weeks.Food intake and water intake were recorded in each group.The levels of Alanine aminotransferase(ALT)and Aspartate aminotransferase(AST)in the serum were measured.ROS and Glutathione(GSH)activity in liver and kidney tissues and urinary protein content were measured.Liver and kidney tissue sections were stained with Hematoxylin-Eosin(HE),and the pathological morphology was observed under a light microscope.The protein levels of HIF-1α,VEGF,and glutathione peroxidase 4(GPX4)in liver and kidney tissues were detected by Western blot.In db/db mice,Fer-1(1 mg·kg-1,ig)could significantly reduce the a-mount of food and water intake,the levels of ALT and AST in serum,the ROS production in liver and kidney tissues,and the level of urine protein,but significantly increase the activity of GSH,thus improve the pathological conditions of liver and kidney.Fer-1 also significantly inhibited HIF-1α and VEGF pro-tein indexes and increased GPX4 protein levels in liver and kidney tissues.Although Fer-1 can not change the body weight and reduce blood glucose in diabetic mice,it can play a therapeutic role in the liver and kidney tissues of diabetic mice in the middle and late stages,and its mechanism may be related to HIF-1α/VEGF and GPX4.
9.Screening and Identification of lncRNA Related to Adipocity of Bone Marrow Microenvironment in Aplastic Anemia
Lei LIU ; Huan-Huan ZHANG ; Xian-Ning ZHANG ; Lu-Lu LIU ; Ming-Tai CHEN
Journal of Experimental Hematology 2024;32(2):610-616
Objective:To systematically screen and identify long noncoding RNA(lncRNA)associated with bone marrow adiposity changes in aplastic anemia(AA).Methods:The PPARγ and C/EBPα ChIP-Seq data in ChIPBase was analyzed by bioinformatics and the potential lncRNA co-transcriptionally regulated by PPARγ and C/EBPα was screened.The expression of candidate lncRNA was verified by qRT-PCR in the in vitro adipogenic differentiation model of BM-MSC,BM-MSC infected with lenti-shPPARγ and lenti-shC/EBPα as well as clinical BM-MSC samples derived from AA and controls.Results:PPARγ and C/EBPα were significantly highly expressed in AA BM-MSC,and knock-down of PPARγ and C/EBPα impaired the adipogenic capacity of AA BM-MSC.PPARγ and C/EBPα cotranscriptionally activate LINC01230 promoter activity in binding sites dependant manner.The LINC01230 was also aberrantly highly expressed in AA BM-MSC compared with controls.Conclusion:PPARγ and C/EBPα are aberrantly expressed in AA BM-MSC and may promote the adipogenic differentiation of AA BM-MSC,and to a certain extent mediate the bone marrow adiposity alteration by transcriptionally activating LINC01230 expression.
10.Dynamic changes of neuronal cells at different time points following cerebral ischemia-reperfusion injury in rats
Xu-Huan ZOU ; Rui LAN ; Xue-Qin FU ; Wei-Wei WANG ; Man-Man WANG ; Chen TANG ; Shuang LIU ; Hong-Yu LI ; Xiao-Ming SHEN
Chinese Pharmacological Bulletin 2024;40(6):1056-1066
Aim To investigate the dynamic changes of neuronal cells at different time points following acute cerebral ischemia-reperfusion injury by establishing a model of brain ischemia-reperfusion injury.Methods Thirty male Sprague-Dawley(SD)rats were ran-domly divided into six groups:sham group and cere-bral ischemia-reperfusion injury(IR)groups at differ-ent time points.Focal cerebral ischemia-reperfusion injury model was established using the middle cerebral artery occlusion(MCAO)technique.The Longa sco-ring method was used to assess neurobehavioral scores in rats.After successful model preparation,routine paraffin sections were made,and TUNEL staining and immunohistochemistry staining with NeuN antibody were performed to observe cell apoptosis and neuronal cell survival,respectively.Immunohistochemistry stai-ning was also performed to investigate the changes in glial fibrillary acidic protein(GFAP)as a marker for astrocytes,ionized calcium-binding adapter molecule 1(IBA-1)as a marker for microglia,and CD31 as a marker for endothelial cells at different time points.Results No significant changes were observed in neu-ronal cells of the sham group at different time points.In the cerebral ischemia-reperfusion injury groups,cell apoptosis was activated at IR3h and increased in quan-tity with morphological damage as time progressed.Ne-uN+neurons showed signs of ischemic injury after IR3h,with abnormal cell morphology.From 12 h,Ne-uN+neurons decreased in a time-dependent manner and reached their peak severity at 24 h.GFAP+astro-cytes decreased significantly after IR3h,while poorly labeled GFAP+astrocytes increased at IR 6 h and al-most disappeared in the infarcted area at 24 h and 48 h.The number of IBA-1+microglia-positive cells de-creased at IR3h,and their volume increased at IR6h.Microglial cell death was observed in the infarcted area at IR12h.CD31+endothelial cells around the infarc-ted cortex and striatum increased significantly after IR3h and persisted until 48 h.Conclusions After cerebral ischemia-reperfusion injury,the number of ap-optotic cells increases with the prolongation of time,and NeuN+neurons exhibit the most severe damage at 24 h.GFAP+astrocytes and microglial cells gradually die over time.The number of CD31+endothelial cells increases significantly around the infarcted cortex and striatum after 3 h of reperfusion and persists until 48 h.


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