1.Effects of electroacupuncture on mitophagy in hippocampal neurons of mouse depression model
Zhiyun YU ; Wenmiao YANG ; Mingmin XU ; Zhiyan LI ; Chenchen YANG ; Junhan JIANG ; Yinan FENG ; Yu GUO
Journal of Beijing University of Traditional Chinese Medicine 2025;48(9):1310-1323
Objective To explore the therapeutic effect and mechanism of electroacupuncture on depressive model mice based on the phosphatase and tensin homolog-induced kinase 1(PINK1)/Parkin pathway.Methods Specific pathogen-free grade male C57BL/6J mice were used.For experiment 1,60 mice were randomly divided into blank,model,sham electroacupuncture,and electroacupuncture groups using the random number table method,with 15 rats per group.For experiment 2,30 mice were randomly divided into normal,cyclosporine A(CsA),and electroacupuncture+CsA groups using the same method,with 10 rats per group.The chronic restraint stress(CRS)was used to establish a depression model.After successful modeling,CRS was continued to maintain model stability.After modeling,1 h before daily CRS stimulation,the electroacupuncture and electroacupuncture+CsA groups received electroacupuncture interventions at the"Baihui"(GV20)and"Zusanli"(ST36)acupoints,using continuous wave stimulation at a frequency of 2 Hz and an intensity of 1 mA for 20 min,once daily for 7 consecutive days.Mice in the sham electroacupuncture group received superficial needling at non-meridian,non-acupoint locations under the axilla 1 h before CRS,with the electroacupuncture device connected but not powered on once a day for 7 consecutive days.Mice in the CsA and electroacupuncture+CsA groups received an intraperitoneal injection of CsA solution(0.2 mg/g)30 min before electroacupuncture intervention,once daily for 7 consecutive days.In experiment 1,depressive-like behavior was assessed using the open field,tail suspension,and sucrose preference tests.The spontaneous excitatory postsynaptic currents(sEPSC)and spontaneous inhibitory postsynaptic currents(sIPSC)parameters of hippocampal neurons were evaluated using brain slice patch clamp techniques.Western blotting was conducted to measure the expression levels of mitochondrial autophagy-related proteins PINK 1,phosphorylated PINK1(p-PINK1),Parkin,microtubule-associated protein 1A/1B-light chain 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ),ubiquitin-binding protein(p62),and mitochondrial markers,including translocase of outer mitochondrial membrane 20(TOMM20),heat shock protein 60(HSP 60),and cytochrome c oxidase Ⅳ(COX Ⅳ).Immunofluorescence was used to detect PINK1 protein expression in the CA3 region of the hippocampus.Transmission electron microscopy was used to examine the ultrastructure of mitochondria in hippocampal neurons.On the basis of experiment 1,experiment 2 evaluated depressive-like behavior in mice using sucrose preference,open field,and tail suspension tests;Western blotting was used to detect the expressions of PINK1,p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,p62,TOMM20,HSP 60,and COX Ⅳ proteins of hippocampus in mice.The mitochondrial ultrastructure was observed in hippocampal neurons using transmission electron microscopy.Results In experiment 1,compared with the blank group,the model and sham electroacupuncture groups exhibited a decrease in sucrose consumption rate,a decrease in the time spent in the center area,a reduced proportion of distance moved in the center area,and an increase in immobility time of tail suspension(P<0.05).The sEPSC and sIPSC in hippocampal neurons decreased in both amplitude and frequency(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus were reduced,whereas the p62 expression level was increased(P<0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus decreased(P<0.05).The number of healthy mitochondria in hippocampal neurons was reduced,with numerous damaged mitochondrial structures observed.Compared to the model and sham electroacupuncture groups,the electroacupuncture group showed an increased in the time spent in the center area,a higher proportion of distance moved in the center area,and an elevated sucrose consumption rate.In contrast,the immobility time in the tail suspension test decreased(P<0.05),whereas the amplitude and frequency of sEPSC and sIPSC in hippocampal neurons increased(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus increased,whereas the p62 expression level decreased(P<0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus increased(P<0.05).Additionally,mitochondrial damage in hippocampal neurons was alleviated,and a notable presence of autophagosomes mitophagy lysosomes was observed.In experiment 2,compared with the normal group,the mice in the CsA and electroacupuncture+CsA groups showed a decrease in the time spent in the center area and the proportion of distance moved in the center area,a decrease in sucrose consumption rate,and an increase in the immobility time in the tail suspension test(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COX Ⅳ expression levels in the hippocampus decreased,whereas p62 expression increased(P<0.05).Many damaged mitochondria were observed in hippocampal neurons.Conclusion Electroacupuncture may exert its antidepressant effects by promoting PINK1/Parkin pathway-mediated mitophagy to eliminate damaged mitochondria,thereby restoring the function of hippocampal neurons in depressive model mice.
2.Effects of electroacupuncture on mitophagy in hippocampal neurons of mouse depression model
Zhiyun YU ; Wenmiao YANG ; Mingmin XU ; Zhiyan LI ; Chenchen YANG ; Junhan JIANG ; Yinan FENG ; Yu GUO
Journal of Beijing University of Traditional Chinese Medicine 2025;48(9):1310-1323
Objective To explore the therapeutic effect and mechanism of electroacupuncture on depressive model mice based on the phosphatase and tensin homolog-induced kinase 1(PINK1)/Parkin pathway.Methods Specific pathogen-free grade male C57BL/6J mice were used.For experiment 1,60 mice were randomly divided into blank,model,sham electroacupuncture,and electroacupuncture groups using the random number table method,with 15 rats per group.For experiment 2,30 mice were randomly divided into normal,cyclosporine A(CsA),and electroacupuncture+CsA groups using the same method,with 10 rats per group.The chronic restraint stress(CRS)was used to establish a depression model.After successful modeling,CRS was continued to maintain model stability.After modeling,1 h before daily CRS stimulation,the electroacupuncture and electroacupuncture+CsA groups received electroacupuncture interventions at the"Baihui"(GV20)and"Zusanli"(ST36)acupoints,using continuous wave stimulation at a frequency of 2 Hz and an intensity of 1 mA for 20 min,once daily for 7 consecutive days.Mice in the sham electroacupuncture group received superficial needling at non-meridian,non-acupoint locations under the axilla 1 h before CRS,with the electroacupuncture device connected but not powered on once a day for 7 consecutive days.Mice in the CsA and electroacupuncture+CsA groups received an intraperitoneal injection of CsA solution(0.2 mg/g)30 min before electroacupuncture intervention,once daily for 7 consecutive days.In experiment 1,depressive-like behavior was assessed using the open field,tail suspension,and sucrose preference tests.The spontaneous excitatory postsynaptic currents(sEPSC)and spontaneous inhibitory postsynaptic currents(sIPSC)parameters of hippocampal neurons were evaluated using brain slice patch clamp techniques.Western blotting was conducted to measure the expression levels of mitochondrial autophagy-related proteins PINK 1,phosphorylated PINK1(p-PINK1),Parkin,microtubule-associated protein 1A/1B-light chain 3-Ⅱ/Ⅰ(LC3-Ⅱ/Ⅰ),ubiquitin-binding protein(p62),and mitochondrial markers,including translocase of outer mitochondrial membrane 20(TOMM20),heat shock protein 60(HSP 60),and cytochrome c oxidase Ⅳ(COX Ⅳ).Immunofluorescence was used to detect PINK1 protein expression in the CA3 region of the hippocampus.Transmission electron microscopy was used to examine the ultrastructure of mitochondria in hippocampal neurons.On the basis of experiment 1,experiment 2 evaluated depressive-like behavior in mice using sucrose preference,open field,and tail suspension tests;Western blotting was used to detect the expressions of PINK1,p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,p62,TOMM20,HSP 60,and COX Ⅳ proteins of hippocampus in mice.The mitochondrial ultrastructure was observed in hippocampal neurons using transmission electron microscopy.Results In experiment 1,compared with the blank group,the model and sham electroacupuncture groups exhibited a decrease in sucrose consumption rate,a decrease in the time spent in the center area,a reduced proportion of distance moved in the center area,and an increase in immobility time of tail suspension(P<0.05).The sEPSC and sIPSC in hippocampal neurons decreased in both amplitude and frequency(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus were reduced,whereas the p62 expression level was increased(P<0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus decreased(P<0.05).The number of healthy mitochondria in hippocampal neurons was reduced,with numerous damaged mitochondrial structures observed.Compared to the model and sham electroacupuncture groups,the electroacupuncture group showed an increased in the time spent in the center area,a higher proportion of distance moved in the center area,and an elevated sucrose consumption rate.In contrast,the immobility time in the tail suspension test decreased(P<0.05),whereas the amplitude and frequency of sEPSC and sIPSC in hippocampal neurons increased(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COXⅣ expression levels in the hippocampus increased,whereas the p62 expression level decreased(P<0.05).The average fluorescence intensity of PINK1 in the CA3 region of the hippocampus increased(P<0.05).Additionally,mitochondrial damage in hippocampal neurons was alleviated,and a notable presence of autophagosomes mitophagy lysosomes was observed.In experiment 2,compared with the normal group,the mice in the CsA and electroacupuncture+CsA groups showed a decrease in the time spent in the center area and the proportion of distance moved in the center area,a decrease in sucrose consumption rate,and an increase in the immobility time in the tail suspension test(P<0.05).p-PINK1,Parkin,LC3-Ⅱ/Ⅰ,TOMM20,HSP 60,and COX Ⅳ expression levels in the hippocampus decreased,whereas p62 expression increased(P<0.05).Many damaged mitochondria were observed in hippocampal neurons.Conclusion Electroacupuncture may exert its antidepressant effects by promoting PINK1/Parkin pathway-mediated mitophagy to eliminate damaged mitochondria,thereby restoring the function of hippocampal neurons in depressive model mice.
3. Nasal continuous positive airway pressure ventilation in children with community-acquired pneumonia under five years of age: a prospective, multi-center clinical study
Jun LIU ; Quan WANG ; Suyun QIAN ; Wenmiao XU ; Lihong LI ; Limin NING ; Xiaoxu REN ; Fang LYU ; Yibing CHENG ; Liujiong GAO ; Chunfeng LIU ; Wei XU ; Liang PEI ; Guoping LU ; Weiming CHEN
Chinese Journal of Pediatrics 2017;55(5):329-333
Objective:
To analyze the clinical characteristics of community-acquired pneumonia (CAP) in children under five years of age and analyze the safety and efficiency of nasal continuous positive airway pressure (NCPAP) ventilation for CAP in this population.
Method:
This was a prospective multicenter study. Children who were admitted to these six centers with CAP and met the NCPAP ventilation indications, aged from 29 d to 5 years, were continuously included during November 2013 to October 2015. The baseline data were collected and NCPAP ventilation were then followed up by operation standards, and the vital signs and arterial blood gas change at special time points were observed and recorded. Any side effect associated with NCPAP were recorded. For categorical variables, comparisons were performed using Fisher test. Rank-sum test and
4.Association of rs1013940 polymorphism in SLC5A7 with Tourette syndrome in Chinese Han popula-tion
Wenmiao LIU ; Aiqin LI ; Yinglei XU ; Mingji YI ; Shiguo LIU ; Hongzai GUAN
Chinese Journal of Behavioral Medicine and Brain Science 2016;25(3):231-234
Objective To investigate the association between rs1013940 in SLC5A7 and Tourette Syndrome ( TS) in Chinese Han population.Methods Polymorphism was genotyped in 401 TS nuclear fam-ilies trios from china by real-time fluorescent quantitive PCR.Transmission disequilibrium test ( TDT) and Haplotype relative risk ( HRR ) were used to analyze the association between the genetic distrbution of rs1013940 and TS and the results were verified by haplotype-based haplotype relative risk( HHRR) .Results No transmission disequilibrium was found between rs1013940 in SLC5A7 and TS by TDT and HRR( TDT:χ2=0.268, P=0.657, OR=0.728,95%CI=0.366-1.451;HRR:χ2=0.111, P=0.739, OR=0.959,95%CI=0.762-1.466) .HHRR also indicated the same result ( HHRR:χ2=0.276, P=0.599, OR=1.082,95%CI=0.806-1.453) .Conclusion The result reveals that there is no significant association between rs1013940 in SLC5A7 and TS in Chinese Han population.However,the results need to be further validated in different populations.
5.Problems and reflection on English teaching in department of gynecology and obstetrics for foreign students
Zhangye XU ; Feng ZHAO ; Ying BAO ; Wenmiao ZHANG ; Yunqin CHEN ; Yinping HUANG
Chinese Journal of Medical Education Research 2013;(8):761-763
Six years' English teaching in department of obstetrics and gynecology in Wenzhou Medical College were reviewed. Language and cultural differences are the main reasons hindering teaching quality. Rational use of a variety of teachers,preparing for lessons adequately,adoption of English image data,supplying and revising English teaching materials,using network auxiliary teaching and forming ef-fective education mode are conducive to improving English teaching quality in department of obstetrics and gynecology for foreign students.
6.Expression of RECK Gene in Placentas from Patients with Preeclampsia and Its Correlation with MMP-2 Activation
Junhong GUO ; Liang XU ; Wenmiao ZHANG ; Yunqin CHEN ; Jing ZHOU ; Xiaohong CHENG ; Yingping HUANG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2009;38(6):818-820,831
Objective To investigate the expression of RECK gene in placentas of patients with preeclampsia and its correlation with MMP-2 activation,and explore the possible roles of RECK gene in the placental trophoblast invasion mechanism.Methods RT-PCR and Western blot were used to detect the expression of RECK mRNA and protein respectively in the placental tissues of normal late pregnant women (normal pregnant group,22 cases) and pre-eclamptic patients(22 mild cases and 20 severe cases).Gelatinase zymography was used to determine MMP-2 activation ratio.Results The expression levels of RECK mRNA and protein from placenta tissues in mild,severe pre-eclamptic group were both significantly higher than those in nomal pregnant group.Moreover,the expression levels of RECK mRNA and protein in severe pre-eclamptic group were obviously increased as compared with those in mild pre-eclamptic group.There was significant difference among the three groups (all P<0.01).MMP-2 activation ratio in mild,severe pre-eclamptic group was significantly lower than that in normal pregnant group.MMP-2 activation ratio in severe pre-eclamptie groups was obviously reduced as compared with mild pre-eclamptic group.There was significant difference among the three groups(all P<0.01).The expression leVels of RECK mRNA and protein were significantly negatively correlated with MMP-2 activation ratio (both P<0.01).Conclusion The abnormal high expression of RECK and inhibition of MMP-2 activation in placentas of pre-eclamptic patients may participate in the process of placental trophoblast shallow invasion.

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