1.Study on the apoptosis-inducing effect of esculetin on acute myeloid leukemia HL-60 cells via regulating the AKT/SKP2/MTH1 pathway
Weihua SONG ; Fuying CHU ; Wei XIE ; Jinliang CHEN ; Ping ZHAO ; Hong QIU ; Jian TAO ; Xiang CHEN
China Pharmacy 2026;37(1):36-41
OBJECTIVE To investigate the apoptosis-inducing effect of esculetin (Esc) on acute myeloid leukemia (AML) HL-60 cells by regulating the protein kinase B (AKT)/S-phase kinase-associated protein 2 (SKP2)/MutT homolog 1 (MTH1) pathway. METHODS AML HL-60 cells were randomly divided into control group (routine culture), Esc low-concentration group (L-Esc group, 25 μmol/L Esc), Esc medium-concentration group (M-Esc group, 50 μmol/L Esc), Esc high-concentration group (H-Esc group, 100 μmol/L Esc), and high-concentration of Esc+ SC79 (AKT agonist) group (100 μmol/L Esc+5 μmol/L SC79). Cell proliferation in each group was detected by MTT assay and colony formation assay. The level of reactive oxygen species (ROS) in cells was measured by using the CM-H2DCFDA fluorescent probe. Cell apoptosis was analyzed by flow cytometry. Western blot assay was performed to detect the expression levels of apoptosis-related proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved caspase-3], AKT/SKP2/MTH1 pathway-related proteins (p-AKT, AKT, SKP2, MTH1), along with the upstream and downstream proteins of AKT phosphatidylinositol 3-kinase (PI3K), cyclin-dependent kinase inhibitor 1 (P21) and cyclin-dependent kinase inhibitor 1B (P27). RESULTS Compared with control group, the cell viability, colony number, and the phosphorylation levels of AKT and PI3K proteins as well as protein expressions of SKP2, MTH1 and Bcl-2 were significantly decreased (P<0.05), while ROS level, apoptosis rate, and the expression levels of Bax, cleaved caspase-3, P21 and P27 proteins were significantly increased (P<0.05). Moreover, the effects of Esc exhibited concentration-dependence (P<0.05). Compared with H-Esc group, above indexes of high-concentration of Esc+ SC79 group were reversed significantly (P<0.05). CONCLUSIONS Esc may promote massive ROS production and induce activation of apoptosis in HL-60 cells by inhibiting the AKT/SKP2/MTH1 pathway, thus inhibiting the proliferation of HL-60 cells.
2.Zishen Huoxue Prescription Alleviates Endoplasmic Reticulum Stress in Hippocampal Neurons of 2-VO Rats via GRP78/PERK/ATF4 Signaling Pathway
Yao SU ; Feng QIU ; Tao YI ; Hanquan LI ; Le XIE ; Xiuli ZHANG ; Dahua WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):93-102
ObjectiveTo investigate the mechanism by which the Zishen Huoxue prescription (ZSHXP) ameliorates cognitive dysfunction in rats with vascular dementia (VD) induced by the bilateral common carotid artery ligation (2-VO model rats) through regulating the glucose-regulated protein 78 (GRP78)/protein kinase R-like endoplasmic reticulum kinase (PERK)/activating transcription factor 4 (ATF4) signaling pathway. MethodsA VD rat model was established via the 2-VO method. A total of 72 male Sprague-Dawley (SD) rats were randomly divided into six groups: Sham group, Model group, donepezil hydrochloride group (0.45 mg·kg-1), and ZSHXP groups at low (8.90 g·kg-1), medium (17.80 g·kg-1), and high (35.60 g·kg-1) doses,with 12 rats in each group. The Morris Water Maze test was utilized to assess spatial learning and memory abilities of rats, and the Novel Object Recognition test was used to evaluate cognitive performance. Hematoxylin-eosin (HE) and Nissl staining were applied to observe the histological and morphological changes in hippocampal tissues. Transmission electron microscopy (TEM) was used to observe the morphological changes of endoplasmic reticulum in rat hippocampal neurons. Immunofluorescence staining was adopted to detect the colocalization of neuronal nuclei antigen (NeuN) with GRP78 and βⅢ Tubulin with gasdermin D (GSDMD) in hippocampal neurons. Western blot was used to detect the expression levels of endoplasmic reticulum stress (ERS)-related proteins including GRP78, PERK, ATF4, phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), C/EBP homologous protein (CHOP), NOD-like receptor protein 3 (NLRP3), Caspase-1 and GSDMD. ResultsCompared with the sham operation group, the model group showed a significantly prolonged escape latency (P<0.01), a significant decrease in the number of platform crossings and the residence time in the target quadrant (P<0.01), and a markedly reduced recognition index (P<0.01). Histological observations revealed that the hippocampal neurons in the model group were disorderly arranged with reduced quantity, deformed and shrunken cell bodies, and pyknotic and hyperchromatic nuclei. The number of Nissl bodies decreased significantly. The number of endoplasmic reticula reduced obviously, accompanied by abnormal dilation and swelling, and the loss of normal folding structure. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly increased in the model group. The protein expression levels of GRP78, p-PERK/PERK, ATF4, CHOP, NLRP3, GSDMD and Caspase-1 in the model group were significantly elevated (P<0.01). Compared with the model group, the donepezil hydrochloride group and the ZSHXP medium- and high-dose groups had a significantly shortened escape latency (P<0.01) and an increased number of platform crossings (P<0.05, P<0.01). The residence time in the target quadrant was increased in the donepezil hydrochloride group and all ZSHXP groups (P<0.05, P<0.01), with a significantly improved recognition index (P<0.01). In the donepezil hydrochloride group and all ZSHXP groups, the number of hippocampal neurons increased with a more compact arrangement and reduced nuclear hyperchromasia. The number of Nissl bodies increased with morphological structures tending to be normal. In the ZSHXP high-dose group, the number of endoplasmic reticula increased and the folding structure was restored. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly weakened in the treatment groups. In the donepezil hydrochloride group, the protein expressions of GRP78, ATF4 and CHOP were increased (P<0.01), while the expression of p-PERK/PERK was decreased (P<0.05). In the ZSHXP low-dose group, the expressions of GRP78, p-PERK/PERK and CHOP were elevated (P<0.05, P<0.01). The ZSHXP medium- and high-dose groups showed a significant decrease in the protein expressions of p-PERK/PERK, ATF4 and CHOP (P<0.01), and the high-dose group had a markedly reduced GRP78 protein expression (P<0.01). In the donepezil hydrochloride group, the Caspase-1 protein expression was increased (P<0.01) and the NLRP3 protein expression was decreased (P<0.01). In the ZSHXP low-dose group, the GSDMD expression was elevated (P<0.01) while the NLRP3 protein expression was reduced (P<0.01). After treatment with medium and high doses of ZSHXP, the protein expression levels of NLRP3, GSDMD and Caspase-1 were significantly decreased (P<0.01). ConclusionThe ameliorative effect of ZSHXP on cognitive function in 2-VO model rats may be associated with its regulation of the GRP78/PERK/ATF4 signaling pathway, which ameliorates ERS and inhibits neuronal pyroptosis.
3.Zishen Huoxue Prescription Alleviates Endoplasmic Reticulum Stress in Hippocampal Neurons of 2-VO Rats via GRP78/PERK/ATF4 Signaling Pathway
Yao SU ; Feng QIU ; Tao YI ; Hanquan LI ; Le XIE ; Xiuli ZHANG ; Dahua WU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):93-102
ObjectiveTo investigate the mechanism by which the Zishen Huoxue prescription (ZSHXP) ameliorates cognitive dysfunction in rats with vascular dementia (VD) induced by the bilateral common carotid artery ligation (2-VO model rats) through regulating the glucose-regulated protein 78 (GRP78)/protein kinase R-like endoplasmic reticulum kinase (PERK)/activating transcription factor 4 (ATF4) signaling pathway. MethodsA VD rat model was established via the 2-VO method. A total of 72 male Sprague-Dawley (SD) rats were randomly divided into six groups: Sham group, Model group, donepezil hydrochloride group (0.45 mg·kg-1), and ZSHXP groups at low (8.90 g·kg-1), medium (17.80 g·kg-1), and high (35.60 g·kg-1) doses,with 12 rats in each group. The Morris Water Maze test was utilized to assess spatial learning and memory abilities of rats, and the Novel Object Recognition test was used to evaluate cognitive performance. Hematoxylin-eosin (HE) and Nissl staining were applied to observe the histological and morphological changes in hippocampal tissues. Transmission electron microscopy (TEM) was used to observe the morphological changes of endoplasmic reticulum in rat hippocampal neurons. Immunofluorescence staining was adopted to detect the colocalization of neuronal nuclei antigen (NeuN) with GRP78 and βⅢ Tubulin with gasdermin D (GSDMD) in hippocampal neurons. Western blot was used to detect the expression levels of endoplasmic reticulum stress (ERS)-related proteins including GRP78, PERK, ATF4, phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), C/EBP homologous protein (CHOP), NOD-like receptor protein 3 (NLRP3), Caspase-1 and GSDMD. ResultsCompared with the sham operation group, the model group showed a significantly prolonged escape latency (P<0.01), a significant decrease in the number of platform crossings and the residence time in the target quadrant (P<0.01), and a markedly reduced recognition index (P<0.01). Histological observations revealed that the hippocampal neurons in the model group were disorderly arranged with reduced quantity, deformed and shrunken cell bodies, and pyknotic and hyperchromatic nuclei. The number of Nissl bodies decreased significantly. The number of endoplasmic reticula reduced obviously, accompanied by abnormal dilation and swelling, and the loss of normal folding structure. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly increased in the model group. The protein expression levels of GRP78, p-PERK/PERK, ATF4, CHOP, NLRP3, GSDMD and Caspase-1 in the model group were significantly elevated (P<0.01). Compared with the model group, the donepezil hydrochloride group and the ZSHXP medium- and high-dose groups had a significantly shortened escape latency (P<0.01) and an increased number of platform crossings (P<0.05, P<0.01). The residence time in the target quadrant was increased in the donepezil hydrochloride group and all ZSHXP groups (P<0.05, P<0.01), with a significantly improved recognition index (P<0.01). In the donepezil hydrochloride group and all ZSHXP groups, the number of hippocampal neurons increased with a more compact arrangement and reduced nuclear hyperchromasia. The number of Nissl bodies increased with morphological structures tending to be normal. In the ZSHXP high-dose group, the number of endoplasmic reticula increased and the folding structure was restored. The fluorescence colocalization of NeuN with GRP78 and βⅢ Tubulin with GSDMD in the hippocampus was significantly weakened in the treatment groups. In the donepezil hydrochloride group, the protein expressions of GRP78, ATF4 and CHOP were increased (P<0.01), while the expression of p-PERK/PERK was decreased (P<0.05). In the ZSHXP low-dose group, the expressions of GRP78, p-PERK/PERK and CHOP were elevated (P<0.05, P<0.01). The ZSHXP medium- and high-dose groups showed a significant decrease in the protein expressions of p-PERK/PERK, ATF4 and CHOP (P<0.01), and the high-dose group had a markedly reduced GRP78 protein expression (P<0.01). In the donepezil hydrochloride group, the Caspase-1 protein expression was increased (P<0.01) and the NLRP3 protein expression was decreased (P<0.01). In the ZSHXP low-dose group, the GSDMD expression was elevated (P<0.01) while the NLRP3 protein expression was reduced (P<0.01). After treatment with medium and high doses of ZSHXP, the protein expression levels of NLRP3, GSDMD and Caspase-1 were significantly decreased (P<0.01). ConclusionThe ameliorative effect of ZSHXP on cognitive function in 2-VO model rats may be associated with its regulation of the GRP78/PERK/ATF4 signaling pathway, which ameliorates ERS and inhibits neuronal pyroptosis.
4.Theoretical application and suggestions of health communication effect evaluation research
Zicong ZHENG ; Dongying XIE ; Ying ZHANG ; Yangmei HUANG ; Meng WANG ; Wuqi QIU
China Modern Doctor 2025;63(8):79-82
Objective To explore theoretical application of health communication effect evaluation in order to provide support for health communication.Methods Using health communication,health activities,health science popularization,effect evaluation as precise keywords,literature collected by databases from built to May 27,2024 was searched,and literature was screened by removing duplicate literature and reading title and abstract of literature.A total of 121 literatures were selected as research samples.Results Most of literatures did not adopt theoretical framework,and only 33 literatures had theoretical applications,mainly in fields of public health and communication.The most widely used literatures were theory of knowing and believing,Lasswell's 5W mode of communication and theory of persuasion,which were dominated by Western theories and lacked application and innovation of localized theories.Conclusion The future health communication effect evaluation should use more theoretical framework to support research,learn from and integrate theories of different disciplines,and explore localization theory research path.
5.Protective effects of Shuangyi Qushi Tongluo Capsules on dexamethasone-induced osteoporosis in mice
Yi LI ; Jian-bin HE ; Jia-xiu XIE ; Quan-mou LUO ; Dong-mei LI ; Jun-hui HE ; Dong-mei WEI ; Chao WEI ; Hong-cong QIU ; Gui-ning WEI ; Bo WANG
Chinese Traditional Patent Medicine 2025;47(6):1834-1842
AIM To investigate the protective effects of Shuangyi Qushi Tongluo Capsules(Shuangyi Capsules)on Dexamethasone(Dex)induced osteoporosis in mice.METHODS The C57BL/6J mice were randomly divided into the control group,the model group,the Xianling Gubao Capsules group(1.5 g/kg),and the low-dose,moderate-dose,and high-dose Shuangyi Capsules groups(0.6,1.2,and 2.4 g/kg).The mouse model of osteoporosis was induced by 8-week intraperitoneal injection of Dex sodium phosphate injection(5 mg/kg).The mice had their femur osteogenesis observed with hematoxylin and eosin(HE)staining and tartrate-resistant acid phosphatase(TRAP)staining;their serum alkaline phosphatase(ALP)and osteocalcin(BGP)activities detected by ELISA;their femoral mRNA expressions of Col-Ⅰ,OCN,and OPN detected by RT-qPCR;and their femoral protein expressions of OPG and RANKL detected by Western blot.Upon the MC3T3-E1 cells exposed to Dex and Shuangyi Capsules,their viability was evaluated by CCK-8 assay;their mineralization determined by alkaline phosphatase staining and alizarin red staining(ARS);and their intracellular ROS level detected using DCFH-DA probe.RESULTS Compared with the model group,Shuangyi Capsules groups demonstrated improved fracture of femoral trabeculae and reduced number of osteoclasts;increased serum ALP and BGP activities(P<0.05,P<0.01);increased femoral expressions of Col-Ⅰ mRNA and OPG protein(P<0.05,P<0.01);and decreased RANKL protein expression(P<0.05).Compared with the MC3T3-E1 cells stimulated by Dex,those underwent further treatment of Shuangyi Capsules demonstrated increased cell viability and ALP activity(P<0.05,P<0.01);increased mineralization and calcium nodule formation;increased expressions of Col-Ⅰ,OCN,OPN mRNA and OPG protein(P<0.05,P<0.01);decreased RANKL protein expression(P<0.05,P<0.01);and reduced ROS levels.CONCLUSION Shuangyi Capsules ameliorate Dex-induced osteoporosis in mice by suppressing osteoclast overactivation,enhancing osteoblast activity,and stimulating bone formation through modulation of Col-Ⅰ,OCN,OPN mRNA and OPG/RANKL protein levels.
6.Etiological characteristics of children with pneumonia and serum CRP,IL-10 and PCT
Ying QIAN ; Min GONG ; Haiqing XIE ; Huixian QIU
Chinese Journal of Nosocomiology 2025;35(9):1357-1361
OBJECTIVE To observe the etiological characteristics,serum C-reactive protein(CRP),interleukin-10(IL-10),procalcitonin(PCT)and treatment outcomes of the children with pneumonia.METHODS A total of 350 children with pneumonia who were treated in Jinhua People's Hospital from Sep.2021 to Sep.2023 and 30 healthy children matched with age and gender who received physical examination were recruited as the research subjects.The etiological characteristics,serum CRP,IL-6,PCT and treatment outcomes of the children with pneumonia were observed.RESULTS The etiological isolation rate was 74.00%(259/350)among the 350 children,the chil-dren who had viral infection accounted for 39.38%,the children with Mycoplasma pneumoniae(MP)infection 32.82%,the children with bacterial infection 20.08%,the children with mixed infections 7.72%.The viral infec-tion was prevalent in winter and was more common among the children aged between 1 and 3 years old;Myco-plasma pneumoniae infection was more common among the children aged between 3 and 6 years old.The CRP levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(10.53±1.54),(38.11±9.32),(21.07±6.55)and(33.33±8.11)mg/L,respectively,higher than those of the healthy children and the children negative for etiological test.The PCT levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(0.61±0.17),(2.84±0.51),(1.35±0.31)and(2.41±0.46)ng/ml,respectively,higher than those of the healthy children and the children negative for etiological test.The IL-10 levels of the viral infection group,the bacterial infection group,the M.pneumoniae infcection group and the mixed infection group were(23.47±5.12),(26.45±7.15),(25.45±5.03)and(25.29±6.05)pg/ml,respectively,higher than those of the healthy children and the children negative for etiological test(P<0.05).The levels of CRP and PCT of the bacterial infection group and the mixed infection group were higher than those of the M.pneumoniae infcection group,and the level of above indexes of the M.pneumoniae infcection group were higher than those of the viral infection group(P<0.05).Among 259 children who were confirmed positive for etiological test,237 had favorable treatment out-comes,and 22 had poor treatment outcomes.The rate of poor treatment outcomes of the children with viral infec-tion was higher than that of the children with M.pneumoniae infcection(x2=5.930,P=0.015);the rate of poor treatment outcomes of the children with mixed infection was higher than that of the children with M.pneumoniae infcection(x2=9.954,P<0.001).CONCLUSIONS The children with viral infection are dominant among the children with pneumonia,followed by the children with M.pneumoniae infcection.The rate of poor treatment outcomes of the children with viral infection or mixed infection is relatively high,the children with M.pneumoniae infcection generally have favorable treatment outcomes.The children with viral infection have slight increases of peripheral blood CRP and PCT,while the children with bacterial infection and mixed infection have a remarkable rise.
7.Development and implementation of a new operation and management model of"one body,two wings and three drives"in a public hospital in Shenzhen
Yue QIU ; Xiaorou XIE ; Wei JIANG ; Qingshan GENG ; Bin DAI ; Yanhui KUANG ; Yunping WANG ; Jun SUN ; Haokai ZHAI
Modern Hospital 2025;25(5):734-737
Objective To assess the efficacy of an innovative operational management model in enhancing the refined operational management of a public hospital.Methods An innovative operational management model,"One Body,Two Wings and Three Drives",was developed,which involved establishing a systematic operational management system,strengthening per-formance and cost control,and reinforcing the supporting roles of discipline construction,scientific and technological innovation,and smart hospital initiatives.This comprehensive approach aimed to systematically promote hospital operational management re-forms and improve overall efficiency and quality.Results After using this model,the hospital presented continuous improve-ments in operational efficiency and medical quality,with key performance indicators trending positively.Over the past three years,the average annual growth rate of outpatient and emergency service visits reached 6.6%,inpatient service visits increased by 5.7%,and the Case Mix Index(CMI)rose by 0.22 over two consecutive years.Conclusion This model is highly systemat-ic,practical,and policy-compatible,providing a replicable path for the high-quality development of public hospitals.
8.Melatonin Alleviates Spinal Cord Ischemia-reperfusion Injury in Rats by Inhibiting Neuron Pyroptosis
Qiu-ping HE ; Lei XIE ; Hai-ning PENG ; Xiao XIAO ; Teng-bo YU
Progress in Modern Biomedicine 2025;25(13):2081-2091
Objective:To investigate the effects of melatonin(MT)on spinal cord ischemia reperfusion injury(SCIRI)and its possible mechanisms in rats.Methods:Forty-two 6-week-old male Sprague Dawley rats were selected for the study and randomly divided into three groups:① Sham group(Sham group,n=14);② model group(model/SCIRI group,n=14);and ③ treatment group(MT group,n=14).Neurological function analysis was used to assess the motor conditions of rats in each group.Nissl staining and hematoxylin eosin(HE)staining were used to observe the number and morphology of neurons in each group,and TUNEL staining was used to evaluate the occurrence of apoptosis of neurons in each group.The expression levels of NOD-like receptor thermal protein domain-associated protein 3(NLRP3),apoptosis-associated speck-like protein(ASC),gasdermin D(GSDMD),the N-terminal fragment of gasdermin D(GSDMD-N),and cysteine proteinase 1(caspase-1)were evaluated using immunofluorescence,immunohistochemistry staining,and Western blot analysis.Results:Compared with the sham operation group,the neurological function score of the model group was significantly decreased,while the neurological function score of the treatment group was higher than that of the model group(P<0.05).The model group had fewer Nissl corpuscles compared with that in the sham operation group and abnormal neuronal morphology(P<0.05).Compared with the model group,the treatment group had a rise in the number of Nissl corpuscles and restored neuronal morphology(P<0.05).Compared with the Sham group,the number of apoptotic neurons increased in the model group,and the protein expression levels of NLRP3,ASC,GSDMD,and caspase-1 increased(P<0.05).Compared with the model group,The number of apoptotic neurons and the protein expressions of NLRP3,ASC,GSDMD and caspase-1 in the treatment group were significantly decreased(P<0.05).Conclusions:MT may alleviate spinal cord ischemia-reperfusion injury by inhibiting pyroptosis in neurons.
9.Development of a quality evaluation system for"internet+pipeline nursing service"based on three-di-mensional quality structure model
Lin ZHANG ; Can ZHANG ; Xiuyun SUN ; Ping XIE ; Zaiping ZHANG ; Qiu SHENG
Modern Hospital 2025;25(2):209-211,215
Objective To develop a scientifically validated and standardized quality evaluation system for"Internet+Pipeline Nursing Service"based on three-dimensional quality structure model.Methods Through literature retrieval and semi-structured interviews,a framework of quality evaluation system was developed based on a three-dimensional quality structure mod-el.Subsequently,the specific content of the evaluation system was finally determined by two rounds of expert consultation through Delphi method.Results The finalized system comprises 41 indicators,including 13 structural indicators,14 process indicators,and 14 outcome indicators.The effective response rates for both rounds of expert consultation were 100%,with expert authority coefficients of 0.803 and 0.831,and expert coordination coefficients of 0.259 and 0.233,respectively.Conclusion The quali-ty evaluation system of"Internet+Pipeline Nursing Service"based on the three-dimensional quality structure model is both scien-tifically sound and reasonable,and it is worthy of promotion and implementation.
10.Application of Cytb and 12S rRNA in wildlife species identification for forensic science
Dezhi JIANG ; Yaxin ZHANG ; Yu ZANG ; Maolei AN ; Zan ZHANG ; Chengcheng QIU ; Yaoheng JIANG ; Wei SONG ; Hong ZHAO ; Kun XIE ; Jiayi CHEN ; Riga WU
Chinese Journal of Forensic Medicine 2025;40(3):308-311,322
Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.

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