1.Screening of Antidepressant Active Components from Curcumae Rhizoma and Its Mechanism in Regulating Nrf2/GPX4/GSH Pathway
Yonggui SONG ; Delin DUAN ; Meixizi LAI ; Yali LIU ; Zhifu AI ; Genhua ZHU ; Huanhua XU ; Qin ZHENG ; Ming YANG ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):211-221
ObjectiveTo screen and evaluate the antidepressant compounds of Curcumae Rhizoma, and explore its mechanism of regulating the nuclear factor erythroid 2-related factor 2(Nrf2)/glutathione(GSH) peroxidase 4(GPX4)/GSH pathway from an antioxidant perspective. MethodsThe antioxidant activities in vitro of 11 characteristic components from Curcumae Rhizoma, including curcumol, curgerenone, curdione, curzerene, curcumenol, curcumenone, dehydrocurdione, isocurcumenol, furanodienone, furanodiene and zederone, were detected using 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt(ABTS) radical scavenging assays. The depression in Drosophila melanogaster was induced by chronic unpredictable mild stress(CUMS), and W1118 wild-type male D. melanogaster were randomly divided into blank group, model group, curcumol group, curgerenone group, curdione group, curzerene group, curcumenol group,curcumenone group, dehydrocurdione group, isocurcumenol group, furanodienone group, furanodiene group, zederone group and fluoxetine group(10 μmol·L-1). The treatment groups received a dose of 0.1 g·L-1 of 11 characteristic components from Curcumae Rhizoma, while the blank and model groups were administered equivalent volumes of solvent. The sucrose preference test, climbing test and forced swimming test were used to evaluate the behavioral indicators of depression in D. melanogaster. Liquid chromatography-mass spectrometry(LC-MS) was used to detect the levels of 5-hydroxytryptamine(5-HT) and dopamine(DA) in the brain of D. melanogaster, and the entropy weight method was used to comprehensively evaluate neurobehavioral and neurotransmitter indicators, resulting in the identification of the antidepressant active components of Curcumae Rhizoma. In addition, a mouse depression model was established by CUMS, and C57BL/6J mice were randomly divided into blank group, model group, low and high dose groups of curzerene(0.5, 1 mg·kg-1), and fluoxetine group(10 mg·kg-1) to confirm the antidepressant effect of the optimal active ingredient by behavioral analysis. Flow cytometry was used to detect the content of reactive oxygen species(ROS) in the hippocampus of mice from each group. Enzyme-linked immunosorbent assay was used to detect the contents of adenosine triphosphate(ATP), superoxide dismutase(SOD), catalase(CAT) and GSH. Transmission electron microscope(TEM) was used to observe the effect of curzerene on the ultrastructure of mitochondria in hippocampal tissue. Western blot was performed to determine the level of Nrf2 protein, and Nrf2 inhibitor(ML385) was used to verify the relationship between the antidepressant effect of curzerene and regulation of Nrf2. Real time fluorescence quantitative polymerase chain reaction(Real-time PCR) was employed to detect the effect of curzerene on the mRNA expression level of GPX. ResultsIn vitro antioxidant experiments showed that curzerene and curgerenone exhibited the most significant ability to scavenge free radicals, and comprehensive evaluation results of entropy weight method indicated that curzerene stood out as the most promising active component. Compared with the blank group, the model group exhibited a significant decrease in sucrose preference coefficient and the number of times entering the open field center(P<0.01), as well as a significant increase in immobility time in the forced swimming and tail suspension tests(P<0.01), and the ROS content in hippocampus significantly elevated(P<0.01), while the ATP content significantly reduced(P<0.01). In the hippocampal neurons of the model group, mitochondrial cristae were disordered, with vacuolation of the inner membrane and severe damage. Nrf2 protein expression level in the model group was significantly decreased(P<0.05), and the antioxidant enzymes SOD, CAT and GSH contents were also significantly reduced(P<0.05, P<0.01), and the gene expression levels of GPX1, GPX4 and GPX7 were significantly decreased(P<0.01). Compared with the model group, the high-dose group of curzerene showed a significant increase in the sucrose preference coefficient and the number of times entering the open field center(P<0.05), as well as a significant decrease in immobility time in the forced swimming and tail suspension tests(P<0.05, P<0.01). The ROS content in the hippocampus of the high-dose group of curzerene was significantly reduced(P<0.01), while the ATP content was significantly increased(P<0.05). The neuronal mitochondrial damage in the hippocampus of the high-dose group of curzerene was alleviated, and the expression level of Nrf2 protein was significantly increased(P<0.05). The Nrf2 inhibitor ML385 reversed the improvement of curzerene on depressive behaviors in CUMS mice. The GSH content in the hippocampal neurons of the high-dose group of curzerene was significantly increased(P<0.01), while there were no significant differences in SOD and CAT contents. The expression level of GPX4 gene in the hippocampal neurons of the high-dose group of curzerene was significantly increased(P<0.05), while there were no significant differences in other GPX genes. ConclusionCurzerene is the best component with antidepressant activity in Curcumae Rhizoma. It may improve mitochondrial dysfunction to exert its antidepressant effect by regulating Nrf2 and its downstream GPX4/GSH pathway rather than CAT or SOD pathways.
2.Exploring Therapeutic Effect of Yuejuwan on Depressed Mice Based on Lipidomics
Zhentao ZHANG ; Dan SU ; Huizhen LI ; Yonggui SONG ; Huanhua XU ; Meixizi LAI ; Zhifu AI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):85-94
ObjectiveTo investigate the effect of Yuejuwan on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice based on lipidomics, and to explore the potential pathways for improving lipid metabolism to prevent depression. MethodsSeven-week-old C57BL/6 mice were randomly divided into blank group, model group, Yuejuwan group(3.6 g·kg-1) and fluoxetine group(10 mg·kg-1), and chronic unpredictable mild stress(CUMS) was used to establish the depression model. After 3 weeks of modeling, each administration group was gavaged with the corresponding drug solution according to the dose, and mice in the blank and model groups were given an equal volume of deionised water by gavage, one time/d for 2 weeks. After administration, the antidepressant effect of Yuejuwan was evaluated by neurobehavioral indices such as sucrose preference test, open field test, tail suspension test and forced swimming test. An automatic biochemical analyzer was used to measure contents of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in mouse serum. Lipidomic analysis of mouse serum, prefrontal cortex and hippocampus was performed based on ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS), and the expression of mammalian target of rapamycin(mTOR), ribosomal protein S6 kinase(S6K), phosphorylation(p)-mTOR, p-S6K in gastric tissues of mice was detected by Western blot. ResultsCompared with the blank group, mice in the model group exhibited significantly reduced sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly increased(P<0.01), and serum levels of TC, TG, LDL-C, HDL-C, AST and ALT were significantly elevated(P<0.05, P<0.01). Compared with the model group, the Yuejuwan group showed a significant increase in the sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly reduced(P<0.01), and the serum levels of TC, TG, LDL-C, AST and ALT were significantly decreased(P<0.05, P<0.01). Lipidomic analysis revealed that Yuejuwan had a significant effect on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice, and The differential lipid metabolites were mainly enriched in the metabolic pathways of glycerophospholipid metabolism, sphingolipid signaling, and glycosylphosphatidylinositol-anchored protein biosynthesis, among which the glycerophospholipid metabolic pathway was the most significant. Western blot results showed that compared with the blank group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the model group were significantly increased(P<0.01). In comparison with the model group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the Yuejuwan group were significantly decreased(P<0.01). ConclusionThe intervention of Yuejuwan on lipid metabolism is one of the potential pathways for its antidepressant effect, which may be related to the regulation of mTOR/S6K signaling pathway upstream of lipid metabolism in the gastric tissues.
3.Molecular Biological Mechanism and Transfusion Strategy of a Jk(a-b-) Family.
Xiao-Yan LI ; Qiong-Fei DENG ; Xiao-Li LAI ; Dan-Dan CHEN ; Dan WANG ; Xuan ZENG
Journal of Experimental Hematology 2025;33(3):869-874
OBJECTIVE:
To investigate the molecular mechanism and explore blood transfusion strategies for a proband exhibiting the JK (a-b-) phenotype and anti-JK3 high frequency antigen antibody and her eight family members.
METHODS:
The Kidd blood phenotype and irregular antibodies in a family were identified by serologic tests. Exon 4-11 and intron region of SLC14A1 gene were sequenced by Sanger method.
RESULTS:
The combination of the gene JK*B (c.499A>G,c.512G>A,c.588A>G) and gene JK*B (c.342-1G>A,588A>G) in this family were considered to result in the JK (a-b-) phenotype in two members. The members carrying gene JK*A(c.130G>A,588A>G) all present serological JKa+W. Members carrying gene JK*B (c.499A>G,c.588A>G) all present serological JKb+W, which has not been previously reported to cause antigenic weakening. The proband with JK (a-b-) phenotype produced anti-JK3 antibodies, the hospital formulated a number of blood preparation strategies for the patient and she was discharged after recovery.
CONCLUSION
In this study, the molecular mechanism of JK (a-b-) in this family was identified, the transfusion strategy of rare blood group was established in our institution preliminary, and the necessity of establishing a rare blood group bank was revealed in this region. It is suggested that JK*B (c.499A>G,c.588A>G) may be a new genetic pattern leading to the weakening of Kidd antigenicity, which lays a foundation for the study of population genetics.
Humans
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Blood Transfusion
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Female
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Kidd Blood-Group System/genetics*
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Phenotype
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Pedigree
4.Inhibition of KLK8 promotes pulmonary endothelial repair by restoring the VE-cadherin/Akt/FOXM1 pathway.
Ying ZHAO ; Hui JI ; Feng HAN ; Qing-Feng XU ; Hui ZHANG ; Di LIU ; Juan WEI ; Dan-Hong XU ; Lai JIANG ; Jian-Kui DU ; Ping-Bo XU ; Yu-Jian LIU ; Xiao-Yan ZHU
Journal of Pharmaceutical Analysis 2025;15(4):101153-101153
Image 1.
5.Influence of NOD-like receptor protein 3 inflammasome on the development and progression of nonalcoholic steatohepatitis and the interventional effect of traditional Chinese medicine
Jinxue ZHANG ; Junhong LIU ; Jiale CHEN ; Dan WANG ; Lining SU ; Yajie CHEN ; Xueqian LAI ; Miaolei WANG ; Yajing LI
Journal of Clinical Hepatology 2025;41(11):2365-2371
Nonalcoholic steatohepatitis (NASH) is a chronic liver disease with the main pathological features of hepatic steatosis, inflammatory cell infiltration, and interstitial fibroplasia, and it is an important risk factor for liver fibrosis, liver cirrhosis, and hepatocellular carcinoma. NOD-like receptor protein 3 (NLRP3) inflammasome is the core of innate immunity, and the abnormal activation of NLRP3 inflammasome is closely associated with the development and progression of NASH, which involves multiple links such as inflammatory response and oxidative stress. A large number of studies have shown that the active ingredients of traditional Chinese medicine (TCM) and TCM compound prescriptions can improve oxidative stress, regulate lipid metabolism, and alleviate liver inflammation by regulating NLRP3 inflammasome. TCM treatment applied in clinical practice has achieved a good therapeutic effect, while inflammasome is one of the key pathways or targets for TCM in improving NASH. This article reviews the mechanism of action of NLRP3 inflammasome in NASH and the research advances in TCM intervention of NLRP3 inflammasome, in order to provide ideas for the clinical TCM treatment of NASH, as well as reference targets and research directions for the research and development of new TCM drugs.
6.A new alkaloid from Huperzia serrata and its biological activity
Xiao-ting QIAO ; Xin-xin CHENG ; Dan WANG ; Mei-han LAI ; Jia-qi WANG ; Xiao-min WANG ; Cai-lian RUAN ; Duo CAO
Acta Pharmaceutica Sinica 2024;59(12):3342-3346
The methanol extract of
7.Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients (version 2024)
Yao LU ; Yang LI ; Leiying ZHANG ; Hao TANG ; Huidan JING ; Yaoli WANG ; Xiangzhi JIA ; Li BA ; Maohong BIAN ; Dan CAI ; Hui CAI ; Xiaohong CAI ; Zhanshan ZHA ; Bingyu CHEN ; Daqing CHEN ; Feng CHEN ; Guoan CHEN ; Haiming CHEN ; Jing CHEN ; Min CHEN ; Qing CHEN ; Shu CHEN ; Xi CHEN ; Jinfeng CHENG ; Xiaoling CHU ; Hongwang CUI ; Xin CUI ; Zhen DA ; Ying DAI ; Surong DENG ; Weiqun DONG ; Weimin FAN ; Ke FENG ; Danhui FU ; Yongshui FU ; Qi FU ; Xuemei FU ; Jia GAN ; Xinyu GAN ; Wei GAO ; Huaizheng GONG ; Rong GUI ; Geng GUO ; Ning HAN ; Yiwen HAO ; Wubing HE ; Qiang HONG ; Ruiqin HOU ; Wei HOU ; Jie HU ; Peiyang HU ; Xi HU ; Xiaoyu HU ; Guangbin HUANG ; Jie HUANG ; Xiangyan HUANG ; Yuanshuai HUANG ; Shouyong HUN ; Xuebing JIANG ; Ping JIN ; Dong LAI ; Aiping LE ; Hongmei LI ; Bijuan LI ; Cuiying LI ; Daihong LI ; Haihong LI ; He LI ; Hui LI ; Jianping LI ; Ning LI ; Xiying LI ; Xiangmin LI ; Xiaofei LI ; Xiaojuan LI ; Zhiqiang LI ; Zhongjun LI ; Zunyan LI ; Huaqin LIANG ; Xiaohua LIANG ; Dongfa LIAO ; Qun LIAO ; Yan LIAO ; Jiajin LIN ; Chunxia LIU ; Fenghua LIU ; Peixian LIU ; Tiemei LIU ; Xiaoxin LIU ; Zhiwei LIU ; Zhongdi LIU ; Hua LU ; Jianfeng LUAN ; Jianjun LUO ; Qun LUO ; Dingfeng LYU ; Qi LYU ; Xianping LYU ; Aijun MA ; Liqiang MA ; Shuxuan MA ; Xainjun MA ; Xiaogang MA ; Xiaoli MA ; Guoqing MAO ; Shijie MU ; Shaolin NIE ; Shujuan OUYANG ; Xilin OUYANG ; Chunqiu PAN ; Jian PAN ; Xiaohua PAN ; Lei PENG ; Tao PENG ; Baohua QIAN ; Shu QIAO ; Li QIN ; Ying REN ; Zhaoqi REN ; Ruiming RONG ; Changshan SU ; Mingwei SUN ; Wenwu SUN ; Zhenwei SUN ; Haiping TANG ; Xiaofeng TANG ; Changjiu TANG ; Cuihua TAO ; Zhibin TIAN ; Juan WANG ; Baoyan WANG ; Chunyan WANG ; Gefei WANG ; Haiyan WANG ; Hongjie WANG ; Peng WANG ; Pengli WANG ; Qiushi WANG ; Xiaoning WANG ; Xinhua WANG ; Xuefeng WANG ; Yong WANG ; Yongjun WANG ; Yuanjie WANG ; Zhihua WANG ; Shaojun WEI ; Yaming WEI ; Jianbo WEN ; Jun WEN ; Jiang WU ; Jufeng WU ; Aijun XIA ; Fei XIA ; Rong XIA ; Jue XIE ; Yanchao XING ; Yan XIONG ; Feng XU ; Yongzhu XU ; Yongan XU ; Yonghe YAN ; Beizhan YAN ; Jiang YANG ; Jiangcun YANG ; Jun YANG ; Xinwen YANG ; Yongyi YANG ; Chunyan YAO ; Mingliang YE ; Changlin YIN ; Ming YIN ; Wen YIN ; Lianling YU ; Shuhong YU ; Zebo YU ; Yigang YU ; Anyong YU ; Hong YUAN ; Yi YUAN ; Chan ZHANG ; Jinjun ZHANG ; Jun ZHANG ; Kai ZHANG ; Leibing ZHANG ; Quan ZHANG ; Rongjiang ZHANG ; Sanming ZHANG ; Shengji ZHANG ; Shuo ZHANG ; Wei ZHANG ; Weidong ZHANG ; Xi ZHANG ; Xingwen ZHANG ; Guixi ZHANG ; Xiaojun ZHANG ; Guoqing ZHAO ; Jianpeng ZHAO ; Shuming ZHAO ; Beibei ZHENG ; Shangen ZHENG ; Huayou ZHOU ; Jicheng ZHOU ; Lihong ZHOU ; Mou ZHOU ; Xiaoyu ZHOU ; Xuelian ZHOU ; Yuan ZHOU ; Zheng ZHOU ; Zuhuang ZHOU ; Haiyan ZHU ; Peiyuan ZHU ; Changju ZHU ; Lili ZHU ; Zhengguo WANG ; Jianxin JIANG ; Deqing WANG ; Jiongcai LAN ; Quanli WANG ; Yang YU ; Lianyang ZHANG ; Aiqing WEN
Chinese Journal of Trauma 2024;40(10):865-881
Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.
8.Application of single-sperm sequencing in resolving the carrier status of preimplantation genetic testing for chromosomal structural rearrangements in Robertsonian translocations
Bao-Qiong LIAO ; Li-Dan LAI ; Ru-Tian LIU ; Qi ZHANG ; Wen-Chang LIAN ; Wu-Ming XIE
National Journal of Andrology 2024;30(6):499-506
Objective:To investigate the application value of single-sperm sequencing in resolving the carrier status of preim-plantation genetic testing(PGT)for chromosomal structural rearrangements in Robertsonian translocations.Methods:Haplotypes were constructed by single-sperm isolation combined with single-sperm sequencing for a patient with 45,XY,der(13;14)(q10;q10).Twenty single-sperm samples were isolated by mechanical braking and subjected to whole-genome amplification(WGA),and then the Asian Screening Array(ASA)gene chip was used to detect the 183 708 single nucleotide polymorphisms(SNP)of the WGA products.The single sperm associated with the translocation that could be used as haplotype inference was detected by copy number variation(CNV)sequencing,and the chromosomal haplotypes with normal and Robertsonian translocations were inferred.Three biopsy samples of embryonic trophoblast cells were used as the objects.After whole-genome amplification,high-throughput sequencing was employed to determine the status of the translocation chromosome carried by the embryos.The available blastocysts were selected for transfer,and the amniotic fluid samples were taken at 18 weeks of gestation to confirm whether the fetus carried the pathogenic muta-tion.Results:A total of 6 037 SNP sites were screened by single-sperm sequencing,and 30 sites selected to distinguish normal and translocation haplotypes.Preimplantation haplotype analysis showed that all the three embryos were euploids without Robertsonian translocation chromosome.Genetic testing of amniotic fluid in the second trimester confirmed that the karyotype of the fetus was 46,XN,carrying no Robertsonian translocation chromosome.Conclusion:For male carriers of Robertsonian translocation,single sperm sequencing can be used to screen SNP sites to construct haplotypes for distinguishing normal and Robertsonian translocation em-bryos,and to provide a basis for embryo selection by preimplantation chromosomal structural genetic testing.
9.Combined application of auditory brainstem response and auditory steady-state response in the evaluation of infants with mild sensorineural hearing loss
Qiuya JIANG ; Qiuying XIE ; Yu HUANG ; Chao HUANG ; Hongli LAN ; Maojie LIU ; Dan LAI
The Journal of Practical Medicine 2024;40(16):2305-2310
Objective To investigate the significance of auditory brainstem response(ABR)combined with auditory steady-state response(ASSR)for the assessment of mild sensorineural hearing loss in infants.Methods Data from 114 infants with mild sensorineural hearing loss were retrospectively analyzed,and their ABR and ASSR results were collected for rank sum test and correlation analysis.Results In the rank sum test,the difference in thresholds between tone-burst ABR(Tb-ABR)and ASSR at 0.5,1,2,4 kHz was statistically significant(P<0.05),and they were also correlated at 0.5,1,2,4 kHz(P<0.05),r=0.613,0.569,0.616,0.71.After grouping by gender and ear,there was a correlation between ABR and ASSR at 0.5,1,2,and 4 kHz,male:r=0.61,0.56,0.671,0.774;female:r=0.581,0.558,0.546,0.608;left ear:r=0.61,0.558,0.576,0.715;right ear:r=0.631,0.581,0.662,0.71.And after grouping by age at diagnosis,only infants diagnosed from 7~12 months of age did not correlate at 0.5 kHz and 1 kHz(P>0.05),while the rest of the groups had a good correlation(P<0.05),0~3 months:r=0.686,0.643,0.671,0.742;4~6 months:r=0.671,0.626,0.616,0.693;7~12 months at 2 kHz and 4 kHz:r=0.571,0.706.Conclusion In infants with mild sensorineural hearing loss,ABR and ASSR correlate in assessing hearing thresholds at all frequencies,and the combination of the two tests could provide a more accurate assessment of the subject's true hearing.
10.Preparation of a rat model of diarrheal irritable bowel syndrome induced by an acetic acid enema combined with binding tail-clamping stress
Biyu LAI ; Mengying HONG ; Xing LI ; Yongjia HE ; Yao CHEN ; Xinwu LI ; Jia SHI ; Zihan TIAN ; Dan LI ; Jing NIE ; Chang SHE
Acta Laboratorium Animalis Scientia Sinica 2024;32(3):317-328
Objective To establish an ideal modeling method for diarrhea predominant irritable bowel syndrome(IBS-D)with anxiely and depression in rats,and to provide a basis for the clinical study of IBS-D.Methods 60 rats were used in this study.(1)At first,20 rats were randomly divided into blank,3%acetic acid enema,4%acetic acid enema,and 5%acetic acid enema groups.After the modeling and observation period,the diarrhea status and the degree of colon injury caused by different modeling concentrations were observed by diarrhea related index and colon histopathology.(2)After the optimal modeling concentration was assessed,40 rats were randomly divided into control(a),acetic acid enema(b),acetic acid+binding(c),and acetic acid+binding+tail clip(d)groups and correspondingly treated for 8 days.After the treatments,the general condition,diarrhea-related index,open field test(OFT)score,and colonic histopathology of rats were evaluated.Results(1)Compared with the blank group,the fecal trait score of 4%acetic acid enema group was increased on days 1 to 3 after intervention(P<0.001),and gradually decreased on days 4 to 7 after intervention.After 1 week,there was no significant difference between the fecal trait score and that of the blank group(P>0.05).Body weight was lower(P<0.01),fecal water content was higher(P<0.001).Compared with blank group,body weight of the 5%acetic acid enema group was decreased(P<0.001),the fecal trait score and diarrhea index were increased(P<0.01).No significant difference was found between 3%acetic acid enema and blank groups.The pathological colon tissue showed that,compared with the blank group,the mucosal structure of the 4%acetic acid enema group was complete with a small amount of inflammatory cell infiltration,and the pathological tissue score showed no significant difference(P>0.05),whereas the 5%acetic acid enema had a medium to large amount of inflammatory cell infiltration,and the pathological tissue score was increased(P<0.01).(2)Compared with group a,group b had lower body weight(P<0.001),and higher fecal trait score,fecal water content and diarrhea index(P<0.01).Compared with a and b groups,the body weight of c and d groups was lower(P<0.001),the fecal traits score,fecal water content,and diarrhea index were increased(P<0.01),and the colon running time was decreased(P<0.01).Compared with group c,Fecal water content in group D was higher(P<0.001).In the OFT score,compared with a and b groups,the OFT distance,standing times,and upright times in c and d groups were lower(P<0.05).Compared with c,the OFT distance,standing times,and upright times in d group were lower(P<0.05).The pathological tissue of colon showed that the mucosal structure of the four groups was complete,and there were different degrees of inflammatory cell infiltration.The pathological tissue scores of groups c and d were higher than those of groups a and b(P<0.05).Conclusions The 4%acetic acid concentration is appropriate for IBS-D modeling.After superposition and binding,the IBS-D diarrhea and internal hypersensitivity characteristic state can be better simulated.After superposition of a tail clip,the IBS-D model of liver stagnation and spleen deficiency can be established successfully.

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