The generation of cellular energy in the form of ATP occurs mainly in mitochondria by oxidative phosphorylation.Cytochrome c oxidase(CytOx),the oxygen accepting and rate-limiting step of the respiratory chain,regulate...The generation of cellular energy in the form of ATP occurs mainly in mitochondria by oxidative phosphorylation.Cytochrome c oxidase(CytOx),the oxygen accepting and rate-limiting step of the respiratory chain,regulates the supply of variable ATP demands in cells by“allosteric ATP-inhibition of CytOx.”This mechanism is based on inhibition of oxygen uptake of CytOx at high ATP/ADP ratios and low ferrocytochrome c concentrations in the mitochondrial matrix via cooperative interaction of the two substrate binding sites in dimeric CytOx.The mechanism keeps mitochondrial membrane potentialΔΨm and reactive oxygen species(ROS)formation at low healthy values.Stress signals increase cytosolic calcium leading to Ca^2+-dependent dephosphorylation of CytOx subunit I at the cytosolic side accompanied by switching off the allosteric ATPinhibition and monomerization of CytOx.This is followed by increase ofΔΨm and formation of ROS.A hypothesis is presented suggesting a dynamic change of binding of NDUFA4,originally identified as a subunit of complex I,between monomeric CytOx(active state with highΔΨm,high ROS and low efficiency)and complex I(resting state with lowΔΨm,low ROS and high efficiency).展开更多
The extraction of cytochrome C was carried out by means of phase transfer technique with three different reverse micellar systems, i.e. , a CTAB micellar solution in n butyl alcohol chloroform(volume ratio 4∶1), an A...The extraction of cytochrome C was carried out by means of phase transfer technique with three different reverse micellar systems, i.e. , a CTAB micellar solution in n butyl alcohol chloroform(volume ratio 4∶1), an AOT micellar solution in isooctane and a SDSS D 2EHPA micellar solution in isooctane. The extraction mechanisms were studied. The results show that the extraction mechanisms for the same proteins with different types of reverse micellar systems can be distinct. The extraction of cytochrome C with CTAB and SDSS D 2EHPA reverse micellar systems are carried out according to the mechanism of electrostatic interaction. However, in the extraction of cytochrome C with the AOT reverse micellar system, the electrostatic interaction between the protein and the surfactant is not important.展开更多
Noncovalent interactions between ligands and targeting proteins are essential for understanding molecular mechanisms of proteins.In this work,we investigated the interaction of Cytochrome c(Cyt c)with maltoligosacchar...Noncovalent interactions between ligands and targeting proteins are essential for understanding molecular mechanisms of proteins.In this work,we investigated the interaction of Cytochrome c(Cyt c)with maltoligosaccharides,namely maltose(Mal Ⅱ),maltotriose(Mal Ⅲ),maltotetraose(Mal Ⅳ),maltopentaose(Mal Ⅴ),maltohexaose(Mal Ⅵ)and maltoheptaose(Mal Ⅶ).Using electrospray ionization mass spetrometry(ESI-MS)assay,the 1:1 and 1:2 complexes formed by Cyt c with maltoligosaccharide ligand were observed.The corresponding association constants were calculated according to the deconvoluted spectra.The order of the relative binding affinities of the selected oligosaccharides with Cyt c were as MalⅢ>MalⅣ>MalⅡ>MalⅤ>MalⅥ>MalⅦ.The results indicated that the stability of noncovalent protein complexes was intimately correlated to the molecular structure of bound ligand.The relevant functional groups that could form H-bonds,electrostatic or hydrophobic forces with protein’s amino residues played an important role for the stability of protein complexes.In addition,the steric structure of ligand was also critical for an appropriate interaction with the binding pocket of proteins.展开更多
Objective: To study the effect of pioglitazone on mitochondrial Ca2+ and cytochrome c (cyt c) in early diabetic rats, and to explore its mechanism of protecting kidney. Methods: 72 DM rats were randomly divided into n...Objective: To study the effect of pioglitazone on mitochondrial Ca2+ and cytochrome c (cyt c) in early diabetic rats, and to explore its mechanism of protecting kidney. Methods: 72 DM rats were randomly divided into negative control group (NC group), negative control+ pioglitazone intervention group (NCP group), diabetes group (DM group) and diabetes +pioglitazone intervention group (DMP group). In NCP and DMP groups, pioglitazone was administered to the stomach, blood glucose, renal mass index, 24-h urine protein, glomerular morphologic indice, glomerular base-membrane thickness and other indexes were measured, and the contents of Ca2+ and Cyt C in renal tissue were measured. Results: (1) the renal metabolism index, glomerular morphologic indice, glomerular base-membrane thickness of DM group was significantly higher than that of NC group (P < 0.01). The renal metabolism index, glomerular morphologic indice, glomerular base-membrane thickness of DMP group was significantly lower than that of DM group, the difference between the two groups was significant(P<0.05). (2) in DM group, mitochondrial Ca2+ and cytoplasmic cytc were higher than those in NC group, while mitochondrial cytc was lower than that in control group. There was significant difference between the two groups (P < 0.05). In DMP group, mitochondrial Ca2+ and cytoplasmic cytc were lower than those in DM group, while mitochondrial cytc was higher than that in control group. There was significant difference between the two groups (P <0.05). Conclusion: Pioglitazone treatment can reduce the release of mitochondrial cytochrome C and maintain mitochondrial calcium homeostasis.展开更多
BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C,enhance expression of apoptotic gene bax,inhibit anti-apoptotic gene bcl-2,and activate caspase-3 to apoptosis;Whereas ...BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C,enhance expression of apoptotic gene bax,inhibit anti-apoptotic gene bcl-2,and activate caspase-3 to apoptosis;Whereas inosine can inhibit neuronal apoptosis which is similar to bil-2.OBJECTIVE: To observe the affects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion,and analyze the pathway of its neuroprotective effect.DESIGN: A randomised controlled animal trial.SETTINGS: Department of Neurology,Rongcheng Second People's Hospital;Department of Neruology,Affiliated Union Hospital,Tongji Medical College,Huazhong University of Science and Technology.MATERIALS: Sixty-eight rats,weighing 230-280 g and clean grade,were used.TdT-mediated dUTP-biotin nick end labeling(TUNEL)and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from Wuhan Boster Biological Co.,Ltd;Inosine injection[200mg(2ml)each] from Qingdao First Pharmaceutical Factory.METHODS:The experiment was accomplished in the animal experimental center in Tongji Medical College of Huazhong University of Science and Technology from December 2003 to June 2005.①Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion(MCAO)with a nylon monofilament suture.The successfully induced rats were assigned to inosine group(n=32)and model group(n=32)at random.Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100mg/kg preoperatively.twice a day,7 days in all.The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively.Each group was randomized into ischemia/reperfusion 2,6,12,24 hours,2,3,7 and 14 days subgroups consisted of 4 rats.The other 4 rats were taken as the sham-operated group,the rats were given the same treatment except for not introduced the filament into the external carotid artery stump.and brain tissue was removed at 2 hours of reperfusion. ②In situ hybridization was performed to examine the expression of cytochrome C mRNA while TUNEL staining was made to characterize apoptosis.③The t test was used to compare the difference of measurement data.MAIN OUTCOME MEASURES:①Neuronal apoptosis in the different regions of the ischemic brain tissue;②Expression of cytochrome C mRNA in the different regions at different time points after MCAO.RESULTS:All the 68 rats were involved in the analysis of results.①Neuronal apoptosis:A small number of TUNEL-positive cells were detected in the sham-operated brain and non-ischemic brain.The number of apoptotic cells in the ischemic cortex peaked at 24 hours of reperfusion[(72.00±1.98)cells]and that in the striatum peaked at 2 days[(94.75±3.57)cells],then decreased to the level of sham-operated group at 14 days.Inosine could reduce apoptotic cells from 12 hours to 7 days of reperfusion as compared with the model group (t:6.19-26.67,P<0.01).②Cytochrome C mRNA expression:There was weak expression of cytochrome C mRNA in both sham-operated brain and contralateral brain. Cytochrome C was detected at 2 hours of reperfusion in ischemic brain[(25.75±3.50),(39.75±2.49)cells],and strongly increased to a peak at 12 hours and 24 hours of reperfusion in cortex and striatum[(122.50±6.69), (119.25±5.12)cells], respectively.Furthermore,inosine could significantly decrease cytochrome C expression in cortex at 12 hours to 14 days of reperfusion after ischemic reperfusion and that in striatum at 12 hours to 3 days(t=8.67-43.26,P<0.01).CONCLUSION:Inosine can exert a neuroprotective effect by inhibiting apoptosis and cytochrome C mRNA expression.展开更多
Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell l...Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. PCR product was展开更多
Most nanozyme research is limited to oxidase and peroxidase.Here,we reported the N,P,or S doped carbon nanotubes(CNTs)for enzyme mimics of nicotinamide adenine dinucleotide(NADH)oxidase and cytochrome c(Cyt c)reductas...Most nanozyme research is limited to oxidase and peroxidase.Here,we reported the N,P,or S doped carbon nanotubes(CNTs)for enzyme mimics of nicotinamide adenine dinucleotide(NADH)oxidase and cytochrome c(Cyt c)reductase.Through the doping of N element,the NADH oxidase-like activity of CNTs is highly improved,the maximum initial velocity for N doped CNT(N-CNT)is increased by 4.28 times compared to that before the modification.Through the analysis of NADH oxidation products,we found that biologically active NAD+was produced,the oxygen was selectively reduced to water or hydrogen peroxide,which is consistent with natural NADH oxidase.Furthermore,we found for the first time that carbon nanotubes can promote the transfer of electrons from NADH to Cyt c,thereby can mimic the properties of Cyt c reductase.展开更多
Monodisperse TiO2 nanoparticles and urchin-like hierarchical TiO2 nanospheres assembled with ultrathin quantum nanowires(about 2 nm)have been synthesized by a simple template-free wet chemical method.The morphology,st...Monodisperse TiO2 nanoparticles and urchin-like hierarchical TiO2 nanospheres assembled with ultrathin quantum nanowires(about 2 nm)have been synthesized by a simple template-free wet chemical method.The morphology,structure,and crystallinity of the TiO2 nanomaterials were investigated by field emission scanning electron microscopy(FESEM),X-ray diffraction(XRD),and high resolution transmission electron microscopy(HRTEM).Electrochemical measurements with the hierarchically nanostructured TiO2 nanospheres as an electrode showed much better reversibility for direct electrochemistry of cytochrome c(cyt c)and much higher sensitivity than for an electrode composed of the monodisperse TiO2 nanoparticles.The excellent performance of the hierarchical TiO2 nanospheres may result from a quantum size effect,and their favorable nanostructure(with the presence of an abundance of both uniform macropores and mesopores),excellent structural stability and high specific surface area.The relative ionic strength had significant effect on the direct electrochemistry.Very high ionic strengths relative to cyt c concentration(I/c)induced a conformational change of cyt c on the nanostructure-coated electrode,from the native state to a partially unfolded one in 25 mmol/L phosphate buffer solution(pH 6.8).展开更多
Background:Tumor necrosis factor receptor-associated protein 1(TRAP1)plays a protective effect in hypoxic cardiomyocytes,but the precise mechanisms are not well clarified.The study is aimed to identify the mechanism o...Background:Tumor necrosis factor receptor-associated protein 1(TRAP1)plays a protective effect in hypoxic cardiomyocytes,but the precise mechanisms are not well clarified.The study is aimed to identify the mechanism of TRAP1 on hypoxic damage in cardiomyocytes.Methods:In this study,the effects of TRAP1 and cytochrome c oxidase subunit Ⅱ(COXⅡ)on apoptosis in hypoxia-induced cardiomyocytes were explored using overexpression and knockdown methods separately.Results:Hypoxia induced cardiomyocyte apoptosis,and TRAP1 overexpression notably inhibited apoptosis induced by hypoxia.Conversely,TRAP1 silencing promoted apoptosis in hypoxic cardiomyocytes.Further investigation revealed that the proapoptotic effects caused by the silencing of TRAP1 were prevented by COXⅡ overexpression,whereas COXⅡ knockdown reduced the antiapoptotic function induced by TRAP1 overexpression.Additionally,changes in the release of cytochrome c from mitochondria into the cytosol and the caspase-3 activity in the cytoplasm,as well as reactive oxygen species production,were found to be correlated with the changes in apoptosis.Conclusions:The current study uncovered that TRAP1 regulates hypoxia-induced cardiomyocyte apoptosis through a mitochondria-dependent apoptotic pathway mediated by COXⅡ,in which reactive oxygen species presents as an important component.展开更多
Cytochromes c covalently bind their heme prosthetic groups through thioether bonds between the vinyl groups of the heme and the thiols of a CXXCH motif within the protein.In Gramnegative bacteria,this process is catal...Cytochromes c covalently bind their heme prosthetic groups through thioether bonds between the vinyl groups of the heme and the thiols of a CXXCH motif within the protein.In Gramnegative bacteria,this process is catalyzed by the Ccm(cytochrome c maturation)proteins,also called System I.The Ccm proteins are found in the bacterial inner membrane,but some(CcmE,CcmG,CcmH,and CcmI)also have soluble functional domains on the periplasmic face of the membrane.Elucidation of the mechanisms involved in the transport and relay of heme and the apo-cytochrome from the bacterial cytosol into the periplasm,and their subsequent reaction,has proved challenging due to the fact that most of the proteins involved are membrane-associated,but recent progress in understanding some key components has thrown up some surprises.In this Review,we discuss advances in our understanding of this process arising from a substrate’s point of view and from recent structural information about individual components.展开更多
The genus Ochotona(pikas)is a clade of cold-tolerant lagomorphs that includes many high-elevation species.Pikas offer a unique opportunity to study adaptations and potential limitations of an ecologically important ma...The genus Ochotona(pikas)is a clade of cold-tolerant lagomorphs that includes many high-elevation species.Pikas offer a unique opportunity to study adaptations and potential limitations of an ecologically important mammal to high-elevation hypoxia.We analyzed the evolution of 3 mitochondrial genes encoding the catalytic core of cytochrome c oxidase(COX)in 10 pika species occupying elevations from sea level to 5000 m.COX is an enzyme highly reliant on oxygen and essential for cell function.One amino acid property,the equilibrium constant(ionization of COOH),was found to be under selection in the overall protein complex.We observed a strong relationship between the net value change in this property and the elevation each species occupies,with higher-elevation species having potentially more efficient proteins.We also found evidence of selection in low-elevation species for potentially less efficient COX,perhaps trading efficiency for heat production in the absence of hypoxia.Our results suggest that different pika species may have evolved elevation-specific COX proteins,specialization that may indicate limitations in their ability to shift their elevational ranges in response to future climate change.展开更多
Recently,a study of mimic enzyme has received more attentions.However,the investigation on the oxidoreductase activity of electron mediators in the biological respiratory chain is still rare.Herein,we found that cadmi...Recently,a study of mimic enzyme has received more attentions.However,the investigation on the oxidoreductase activity of electron mediators in the biological respiratory chain is still rare.Herein,we found that cadmium sulfide(CdS)nanorods can catalyze the formation of superoxide anions.Due to the role of the photo-generated holes and the nicotinamide adenine dinucleotide(NADH)oxidation promoted by superoxide anion(O_(2)^(•−)),the CdS exhibits NADH oxidase-like activity and can be coupled with dehydrogenase to realize the recycling of NADH.It is worth mentioning that the bio-electron acceptor,cytochrome c(Cyt c),as a chromogenic substrate,can accept electrons transferred from O_(2)^(•−),which demonstrates the Cyt c reductase-like activity of CdS under physiological pH conditions.For different substrates,O_(2)^(•−)induced from CdS show oxidizing capacity for NADH and reducing capacity for Cyt c,which provides a new perspective for the in-depth study of new nanozyme.展开更多
In order to demonstrate the effect of the sixth axial ligand on the biological funtionof heme proteins, we have determined the formal potentials and electron transfer numbersof imidazole (Im), 1-methylimidazole (1-MeI...In order to demonstrate the effect of the sixth axial ligand on the biological funtionof heme proteins, we have determined the formal potentials and electron transfer numbersof imidazole (Im), 1-methylimidazole (1-MeIm) and 1-ethylimidazole (1-EtIm) complexes ofcytochrome c(cyt c) with an optically transparent thin-layer electrode.展开更多
Human and animal hairs have been used in forensic investigations for over a century.Hair is stable under adverse natural conditions;hence,it is often recovered at the crime scene,and it is necessary to determine wheth...Human and animal hairs have been used in forensic investigations for over a century.Hair is stable under adverse natural conditions;hence,it is often recovered at the crime scene,and it is necessary to determine whether the hair is of human or animal origin.Morphological and genetic characteristics are useful to differentiate human hair from animal hair.In the present study,we analyzed the distinguishing characteristics of hair of various species.In addition,we explore species identification by cytochrome c oxidase I mitochondrial gene analysis.We confirm that both the microscopic and molecular analyses of hairs are useful in forensic investigations.展开更多
RNA analysis offers many potential applications in forensic science,and molecular identification of body fluids by analysis of cell‑specific RNA markers represents a new technique for use in forensic cases.However,due...RNA analysis offers many potential applications in forensic science,and molecular identification of body fluids by analysis of cell‑specific RNA markers represents a new technique for use in forensic cases.However,due to the nature of forensic materials that often admixed with nonhuman cellular components,human‑specific RNA quantification is required for the forensic RNA assays.Quantification assay for human RNA has been developed in the present study with respect to body fluid samples in forensic biology.The quantitative assay is based on real‑time reverse transcription‑polymerase chain reaction of mitochondrial RNA cytochrome c oxidase subunit I and capable of RNA quantification with high reproducibility and a wide dynamic range.The human RNA quantification improves the quality of mRNA profiling in the identification of body fluids of saliva and semen because the quantification assay can exclude the influence of nonhuman components and reduce the adverse affection from degraded RNA fragments.展开更多
Benthodytes tetrapapillata sp.nov.was collected from a seamount located on the Caroline Ridge at a depth of 2289 m,during the cruise of R/V Kexue in June 2019.We provided detailed descriptions of external and deposits...Benthodytes tetrapapillata sp.nov.was collected from a seamount located on the Caroline Ridge at a depth of 2289 m,during the cruise of R/V Kexue in June 2019.We provided detailed descriptions of external and deposits morphology.The phylogenetic analyses based on cytochrome c oxidase I(COI)and a concatenated dataset of 16S and COI genes showed that the new species belonged to Benthodytes that is not monophyletic.Both details of morphological comparisons and molecular analyses confirmed that Benthodytes tetrapapillata sp.nov.is a new psychropotid species.A state of main morphological characters in valid species of Benthodytes is also provided in this study.展开更多
Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan c...Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.展开更多
The sea star Asterias amurensis is widely viewed as a severe“marine pest”because of its broad feeding habits.Over the past few decades,A.amurensis undergoes massive and sporadic population outbreaks worldwide,causin...The sea star Asterias amurensis is widely viewed as a severe“marine pest”because of its broad feeding habits.Over the past few decades,A.amurensis undergoes massive and sporadic population outbreaks worldwide,causing extensive economic and ecological losses to the local aquaculture industry and marine ecosystem.Understanding the genetic diversity and population structure of A.amurensis can provide vital information for resource management.By analyzing the polymorphism of the mitochondrial cytochrome C oxidase subunit I(COI)gene and ten simple sequence repeat(SSR)microsatellites markers,the genetic diversity and population structure of A.amurensis of four populations along the northern coast of China was uncovered.A total of 36 haplotypes were identified,and a main haplotype was found in four populations.The Qingdao(QD)population displayed the highest genetic diversity among all the populations.The AMOVA and pairwise F_(st)showed that there was small but statistically significant population differentiation among the four populations,especially between QD and Weihai(WH).Moreover,the principal component analysis(PCA)and admixture analysis showed that several individuals in Yantai(YT)and Dalian(DL)had little genetic association with other individuals.Overall,this study provided useful information of the genetic diversity and population structure of A.amurensis and will contribute to the resource management of A.amurensis in China.展开更多
OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cy...OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cyt-C)and apoptosis-inducing factor(AIF)in chronic heart failure(CHF)rats.METHODS:Sixty-five rats were randomly divided into normal group(n=10)and model-I group(n=55).After modeling,CHF rats in model-I group were divided into model group,moxibustion group,benazepril group,moxibustion plus benazepril group(abbreviated as aibei group,the same below),10 rats in each group.Echocardiogram index was examined by echocardiography.Hemodynamic indices were measured by rat cardiac function meter.Serum B-type brain natriuretic peptide(BNP)was detected by enzymelinked immunosorbent assay.Myocardial cells apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining.Pathological changes of myocardial tissues were observed by hematoxylin and eosin staining.The expression levels of Cyt-C and AIF in myocardial tissues were detected by Western blot.RESULTS:Compared with normal group,ejection fraction and left ventricular diameter shortening rate in model-Ⅰgroup were significantly reduced,myocardial cells of rats in model group exhibited unclear transverse striations,cells swellings and vacuoles,cardiac functions were deteriorated,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly increased.Compared with model group,myocardial cells of rats in moxibustion group,benazepril group,and aibei group were dyed more evenly,muscle fibers were arranged relatively neatly,cardiac functions were improved,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly decreased.Compared with aibei group,cardiac functions were worsened,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were increased.CONCLUSION:Moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril could improve CHF better than moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)or benazepril alone.The mechanisms might be that they can inhibit the expressions of Cyt-C and AIF,and inhibit the apoptosis of cardiomyocytes.展开更多
文摘The generation of cellular energy in the form of ATP occurs mainly in mitochondria by oxidative phosphorylation.Cytochrome c oxidase(CytOx),the oxygen accepting and rate-limiting step of the respiratory chain,regulates the supply of variable ATP demands in cells by“allosteric ATP-inhibition of CytOx.”This mechanism is based on inhibition of oxygen uptake of CytOx at high ATP/ADP ratios and low ferrocytochrome c concentrations in the mitochondrial matrix via cooperative interaction of the two substrate binding sites in dimeric CytOx.The mechanism keeps mitochondrial membrane potentialΔΨm and reactive oxygen species(ROS)formation at low healthy values.Stress signals increase cytosolic calcium leading to Ca^2+-dependent dephosphorylation of CytOx subunit I at the cytosolic side accompanied by switching off the allosteric ATPinhibition and monomerization of CytOx.This is followed by increase ofΔΨm and formation of ROS.A hypothesis is presented suggesting a dynamic change of binding of NDUFA4,originally identified as a subunit of complex I,between monomeric CytOx(active state with highΔΨm,high ROS and low efficiency)and complex I(resting state with lowΔΨm,low ROS and high efficiency).
文摘The extraction of cytochrome C was carried out by means of phase transfer technique with three different reverse micellar systems, i.e. , a CTAB micellar solution in n butyl alcohol chloroform(volume ratio 4∶1), an AOT micellar solution in isooctane and a SDSS D 2EHPA micellar solution in isooctane. The extraction mechanisms were studied. The results show that the extraction mechanisms for the same proteins with different types of reverse micellar systems can be distinct. The extraction of cytochrome C with CTAB and SDSS D 2EHPA reverse micellar systems are carried out according to the mechanism of electrostatic interaction. However, in the extraction of cytochrome C with the AOT reverse micellar system, the electrostatic interaction between the protein and the surfactant is not important.
基金supported by the National Natural Science Foundation of China(Grant No.21675176)the Natural Science Foundation of Hubei Province(Grant No.2014CFA025)the Preferred Research Foundation for the Returned Overseas Scholars from Ministry of Human Resources and Social Security of the People’s Republic of China(Grant No.BZY14036)for financial supports.
文摘Noncovalent interactions between ligands and targeting proteins are essential for understanding molecular mechanisms of proteins.In this work,we investigated the interaction of Cytochrome c(Cyt c)with maltoligosaccharides,namely maltose(Mal Ⅱ),maltotriose(Mal Ⅲ),maltotetraose(Mal Ⅳ),maltopentaose(Mal Ⅴ),maltohexaose(Mal Ⅵ)and maltoheptaose(Mal Ⅶ).Using electrospray ionization mass spetrometry(ESI-MS)assay,the 1:1 and 1:2 complexes formed by Cyt c with maltoligosaccharide ligand were observed.The corresponding association constants were calculated according to the deconvoluted spectra.The order of the relative binding affinities of the selected oligosaccharides with Cyt c were as MalⅢ>MalⅣ>MalⅡ>MalⅤ>MalⅥ>MalⅦ.The results indicated that the stability of noncovalent protein complexes was intimately correlated to the molecular structure of bound ligand.The relevant functional groups that could form H-bonds,electrostatic or hydrophobic forces with protein’s amino residues played an important role for the stability of protein complexes.In addition,the steric structure of ligand was also critical for an appropriate interaction with the binding pocket of proteins.
基金Key medical and health project of nanjing military region
文摘Objective: To study the effect of pioglitazone on mitochondrial Ca2+ and cytochrome c (cyt c) in early diabetic rats, and to explore its mechanism of protecting kidney. Methods: 72 DM rats were randomly divided into negative control group (NC group), negative control+ pioglitazone intervention group (NCP group), diabetes group (DM group) and diabetes +pioglitazone intervention group (DMP group). In NCP and DMP groups, pioglitazone was administered to the stomach, blood glucose, renal mass index, 24-h urine protein, glomerular morphologic indice, glomerular base-membrane thickness and other indexes were measured, and the contents of Ca2+ and Cyt C in renal tissue were measured. Results: (1) the renal metabolism index, glomerular morphologic indice, glomerular base-membrane thickness of DM group was significantly higher than that of NC group (P < 0.01). The renal metabolism index, glomerular morphologic indice, glomerular base-membrane thickness of DMP group was significantly lower than that of DM group, the difference between the two groups was significant(P<0.05). (2) in DM group, mitochondrial Ca2+ and cytoplasmic cytc were higher than those in NC group, while mitochondrial cytc was lower than that in control group. There was significant difference between the two groups (P < 0.05). In DMP group, mitochondrial Ca2+ and cytoplasmic cytc were lower than those in DM group, while mitochondrial cytc was higher than that in control group. There was significant difference between the two groups (P <0.05). Conclusion: Pioglitazone treatment can reduce the release of mitochondrial cytochrome C and maintain mitochondrial calcium homeostasis.
基金the Natural Science Fund of Shandong Province, No.Y2001C04
文摘BACKGROUND: It has been demonstrated that adenosine can induce glial cell to release cytochrome C,enhance expression of apoptotic gene bax,inhibit anti-apoptotic gene bcl-2,and activate caspase-3 to apoptosis;Whereas inosine can inhibit neuronal apoptosis which is similar to bil-2.OBJECTIVE: To observe the affects of inosine on neuronal apoptosis and expression of cytochrome C mRNA in rats after focal cerebral ischemia/reperfusion,and analyze the pathway of its neuroprotective effect.DESIGN: A randomised controlled animal trial.SETTINGS: Department of Neurology,Rongcheng Second People's Hospital;Department of Neruology,Affiliated Union Hospital,Tongji Medical College,Huazhong University of Science and Technology.MATERIALS: Sixty-eight rats,weighing 230-280 g and clean grade,were used.TdT-mediated dUTP-biotin nick end labeling(TUNEL)and cytochrome C mRNA in situ hybridization kits and DAB staining kit were purchased from Wuhan Boster Biological Co.,Ltd;Inosine injection[200mg(2ml)each] from Qingdao First Pharmaceutical Factory.METHODS:The experiment was accomplished in the animal experimental center in Tongji Medical College of Huazhong University of Science and Technology from December 2003 to June 2005.①Sixty-four rats were made into focal ischemia by middle cerebral artery occlusion(MCAO)with a nylon monofilament suture.The successfully induced rats were assigned to inosine group(n=32)and model group(n=32)at random.Rats in the inosine group were intraperitoneally administrated with inosine in dose of 100mg/kg preoperatively.twice a day,7 days in all.The rats in the control group were injected with the same dose of saline solution by the similar way preoperatively.Each group was randomized into ischemia/reperfusion 2,6,12,24 hours,2,3,7 and 14 days subgroups consisted of 4 rats.The other 4 rats were taken as the sham-operated group,the rats were given the same treatment except for not introduced the filament into the external carotid artery stump.and brain tissue was removed at 2 hours of reperfusion. ②In situ hybridization was performed to examine the expression of cytochrome C mRNA while TUNEL staining was made to characterize apoptosis.③The t test was used to compare the difference of measurement data.MAIN OUTCOME MEASURES:①Neuronal apoptosis in the different regions of the ischemic brain tissue;②Expression of cytochrome C mRNA in the different regions at different time points after MCAO.RESULTS:All the 68 rats were involved in the analysis of results.①Neuronal apoptosis:A small number of TUNEL-positive cells were detected in the sham-operated brain and non-ischemic brain.The number of apoptotic cells in the ischemic cortex peaked at 24 hours of reperfusion[(72.00±1.98)cells]and that in the striatum peaked at 2 days[(94.75±3.57)cells],then decreased to the level of sham-operated group at 14 days.Inosine could reduce apoptotic cells from 12 hours to 7 days of reperfusion as compared with the model group (t:6.19-26.67,P<0.01).②Cytochrome C mRNA expression:There was weak expression of cytochrome C mRNA in both sham-operated brain and contralateral brain. Cytochrome C was detected at 2 hours of reperfusion in ischemic brain[(25.75±3.50),(39.75±2.49)cells],and strongly increased to a peak at 12 hours and 24 hours of reperfusion in cortex and striatum[(122.50±6.69), (119.25±5.12)cells], respectively.Furthermore,inosine could significantly decrease cytochrome C expression in cortex at 12 hours to 14 days of reperfusion after ischemic reperfusion and that in striatum at 12 hours to 3 days(t=8.67-43.26,P<0.01).CONCLUSION:Inosine can exert a neuroprotective effect by inhibiting apoptosis and cytochrome C mRNA expression.
文摘Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase ( COX) activity in mouse Sertoli cell line TM4. Methods The coding region of CoxTa2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. PCR product was
基金the National Key Research and Development Program of China(No.2019YFA0709202)Natural Science Foundation of Jilin Province(No.20220101055JC)+1 种基金the International Cooperation Project of Jilin Scientific and Technological Development Program(No.20190701059GH)the National Natural Science Foundation of China(No.31301177).
文摘Most nanozyme research is limited to oxidase and peroxidase.Here,we reported the N,P,or S doped carbon nanotubes(CNTs)for enzyme mimics of nicotinamide adenine dinucleotide(NADH)oxidase and cytochrome c(Cyt c)reductase.Through the doping of N element,the NADH oxidase-like activity of CNTs is highly improved,the maximum initial velocity for N doped CNT(N-CNT)is increased by 4.28 times compared to that before the modification.Through the analysis of NADH oxidation products,we found that biologically active NAD+was produced,the oxygen was selectively reduced to water or hydrogen peroxide,which is consistent with natural NADH oxidase.Furthermore,we found for the first time that carbon nanotubes can promote the transfer of electrons from NADH to Cyt c,thereby can mimic the properties of Cyt c reductase.
基金This work was supported by the National Basic Research Program of China(No.2010CB934700)the National Natural Science Foundation of China(Nos.20973019,50902007,and 50725208).
文摘Monodisperse TiO2 nanoparticles and urchin-like hierarchical TiO2 nanospheres assembled with ultrathin quantum nanowires(about 2 nm)have been synthesized by a simple template-free wet chemical method.The morphology,structure,and crystallinity of the TiO2 nanomaterials were investigated by field emission scanning electron microscopy(FESEM),X-ray diffraction(XRD),and high resolution transmission electron microscopy(HRTEM).Electrochemical measurements with the hierarchically nanostructured TiO2 nanospheres as an electrode showed much better reversibility for direct electrochemistry of cytochrome c(cyt c)and much higher sensitivity than for an electrode composed of the monodisperse TiO2 nanoparticles.The excellent performance of the hierarchical TiO2 nanospheres may result from a quantum size effect,and their favorable nanostructure(with the presence of an abundance of both uniform macropores and mesopores),excellent structural stability and high specific surface area.The relative ionic strength had significant effect on the direct electrochemistry.Very high ionic strengths relative to cyt c concentration(I/c)induced a conformational change of cyt c on the nanostructure-coated electrode,from the native state to a partially unfolded one in 25 mmol/L phosphate buffer solution(pH 6.8).
基金supported by the National Natural Science Foundation of China(NSFC)(Grant No:81101426,81571898).
文摘Background:Tumor necrosis factor receptor-associated protein 1(TRAP1)plays a protective effect in hypoxic cardiomyocytes,but the precise mechanisms are not well clarified.The study is aimed to identify the mechanism of TRAP1 on hypoxic damage in cardiomyocytes.Methods:In this study,the effects of TRAP1 and cytochrome c oxidase subunit Ⅱ(COXⅡ)on apoptosis in hypoxia-induced cardiomyocytes were explored using overexpression and knockdown methods separately.Results:Hypoxia induced cardiomyocyte apoptosis,and TRAP1 overexpression notably inhibited apoptosis induced by hypoxia.Conversely,TRAP1 silencing promoted apoptosis in hypoxic cardiomyocytes.Further investigation revealed that the proapoptotic effects caused by the silencing of TRAP1 were prevented by COXⅡ overexpression,whereas COXⅡ knockdown reduced the antiapoptotic function induced by TRAP1 overexpression.Additionally,changes in the release of cytochrome c from mitochondria into the cytosol and the caspase-3 activity in the cytoplasm,as well as reactive oxygen species production,were found to be correlated with the changes in apoptosis.Conclusions:The current study uncovered that TRAP1 regulates hypoxia-induced cardiomyocyte apoptosis through a mitochondria-dependent apoptotic pathway mediated by COXⅡ,in which reactive oxygen species presents as an important component.
基金supported in PDB's lab by the BBSRC and an EPSRC studentship to EBS.EBS is currently supported by a Chinese Academy of Sciences Fellowship for Young International Scientists(No.2010Y2SB01)a grant from the National Natural Science Foundation of China(Grant No.31150110150).
文摘Cytochromes c covalently bind their heme prosthetic groups through thioether bonds between the vinyl groups of the heme and the thiols of a CXXCH motif within the protein.In Gramnegative bacteria,this process is catalyzed by the Ccm(cytochrome c maturation)proteins,also called System I.The Ccm proteins are found in the bacterial inner membrane,but some(CcmE,CcmG,CcmH,and CcmI)also have soluble functional domains on the periplasmic face of the membrane.Elucidation of the mechanisms involved in the transport and relay of heme and the apo-cytochrome from the bacterial cytosol into the periplasm,and their subsequent reaction,has proved challenging due to the fact that most of the proteins involved are membrane-associated,but recent progress in understanding some key components has thrown up some surprises.In this Review,we discuss advances in our understanding of this process arising from a substrate’s point of view and from recent structural information about individual components.
文摘The genus Ochotona(pikas)is a clade of cold-tolerant lagomorphs that includes many high-elevation species.Pikas offer a unique opportunity to study adaptations and potential limitations of an ecologically important mammal to high-elevation hypoxia.We analyzed the evolution of 3 mitochondrial genes encoding the catalytic core of cytochrome c oxidase(COX)in 10 pika species occupying elevations from sea level to 5000 m.COX is an enzyme highly reliant on oxygen and essential for cell function.One amino acid property,the equilibrium constant(ionization of COOH),was found to be under selection in the overall protein complex.We observed a strong relationship between the net value change in this property and the elevation each species occupies,with higher-elevation species having potentially more efficient proteins.We also found evidence of selection in low-elevation species for potentially less efficient COX,perhaps trading efficiency for heat production in the absence of hypoxia.Our results suggest that different pika species may have evolved elevation-specific COX proteins,specialization that may indicate limitations in their ability to shift their elevational ranges in response to future climate change.
基金supported by the National Key R&D Program of China(No.2019YFA0709202)MOST China(No.2016YFA0203200)+1 种基金the International Cooperation Project of Jilin Scientific and Technological Development Program(No.20190701059GH)the National Natural Science Foundation of China(Nos.21721003 and 31301177).
文摘Recently,a study of mimic enzyme has received more attentions.However,the investigation on the oxidoreductase activity of electron mediators in the biological respiratory chain is still rare.Herein,we found that cadmium sulfide(CdS)nanorods can catalyze the formation of superoxide anions.Due to the role of the photo-generated holes and the nicotinamide adenine dinucleotide(NADH)oxidation promoted by superoxide anion(O_(2)^(•−)),the CdS exhibits NADH oxidase-like activity and can be coupled with dehydrogenase to realize the recycling of NADH.It is worth mentioning that the bio-electron acceptor,cytochrome c(Cyt c),as a chromogenic substrate,can accept electrons transferred from O_(2)^(•−),which demonstrates the Cyt c reductase-like activity of CdS under physiological pH conditions.For different substrates,O_(2)^(•−)induced from CdS show oxidizing capacity for NADH and reducing capacity for Cyt c,which provides a new perspective for the in-depth study of new nanozyme.
基金Project supported by the National Natural Science Foundation of China.
文摘In order to demonstrate the effect of the sixth axial ligand on the biological funtionof heme proteins, we have determined the formal potentials and electron transfer numbersof imidazole (Im), 1-methylimidazole (1-MeIm) and 1-ethylimidazole (1-EtIm) complexes ofcytochrome c(cyt c) with an optically transparent thin-layer electrode.
文摘Human and animal hairs have been used in forensic investigations for over a century.Hair is stable under adverse natural conditions;hence,it is often recovered at the crime scene,and it is necessary to determine whether the hair is of human or animal origin.Morphological and genetic characteristics are useful to differentiate human hair from animal hair.In the present study,we analyzed the distinguishing characteristics of hair of various species.In addition,we explore species identification by cytochrome c oxidase I mitochondrial gene analysis.We confirm that both the microscopic and molecular analyses of hairs are useful in forensic investigations.
基金The authors would like to thank the support from Beijing Municipal Natural Science Foundation(7,163,221)Ministry of Public Security of Material Evidence Identification Center(2017FGKFKT05)the Program for Young Innovative Research Team in China University of Political Science and Law(2014CXTD04,2016CXTD05).
文摘RNA analysis offers many potential applications in forensic science,and molecular identification of body fluids by analysis of cell‑specific RNA markers represents a new technique for use in forensic cases.However,due to the nature of forensic materials that often admixed with nonhuman cellular components,human‑specific RNA quantification is required for the forensic RNA assays.Quantification assay for human RNA has been developed in the present study with respect to body fluid samples in forensic biology.The quantitative assay is based on real‑time reverse transcription‑polymerase chain reaction of mitochondrial RNA cytochrome c oxidase subunit I and capable of RNA quantification with high reproducibility and a wide dynamic range.The human RNA quantification improves the quality of mRNA profiling in the identification of body fluids of saliva and semen because the quantification assay can exclude the influence of nonhuman components and reduce the adverse affection from degraded RNA fragments.
基金Supported by the National Key Research and Development Program of China (No.2021YFE0193700)the Strategic Priority Research Program of the Chinese Academy of Sciences (No.XDB42000000)+3 种基金the National Natural Science Foundation of China (No.41930533)the Biological Resources ProgramChinese Academy of Sciences (No.KFJ-BRP-017-38)the Major Scientific and Technological Projects of Hainan Province (No.ZDKJ2021036)。
文摘Benthodytes tetrapapillata sp.nov.was collected from a seamount located on the Caroline Ridge at a depth of 2289 m,during the cruise of R/V Kexue in June 2019.We provided detailed descriptions of external and deposits morphology.The phylogenetic analyses based on cytochrome c oxidase I(COI)and a concatenated dataset of 16S and COI genes showed that the new species belonged to Benthodytes that is not monophyletic.Both details of morphological comparisons and molecular analyses confirmed that Benthodytes tetrapapillata sp.nov.is a new psychropotid species.A state of main morphological characters in valid species of Benthodytes is also provided in this study.
基金Supported by the Zhejiang Provincial Key Research and Development Program (No.2021C02047)。
文摘Accurate species identification is a key component of biodiversity research.DNA barcoding is an effective molecular method used for fish species identification.We aimed to study the DNA barcoding of fish in Zhoushan coastal waters,explore the differences and applicability of two gene fragments(12S rRNA and COI)of DNA barcoding in fish species identification,and established a comprehensive fish barcoding reference database.Two hundred and eighty-seven captured fish samples from Zhoushan coastal waters were identified using morphological characteristics and DNA barcoding.A total of 26412S rRNA sequences(belonging to eight orders,31 families,55 genera,and 66 species)and 188 COI sequences(belonging to seven orders,30 families,48 genera,and 58 species)were obtained.The lengths of the 12S rRNA sequences ranged from 165 to 178 bp,and the guanine-cytosine(GC)content was 45.37%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.10%and 26.66%,respectively.The length of the COI sequence ranged 574–655 bp,and the content of GC was 45.97%.The average 12S rRNA interspecific and intraspecific genetic distances(K2P)were 0.16%and 27.45%,respectively.The minimum interspecific genetic distances of 12S rRNA and COI(1.23%and 1.86%)were both greater than their maximum intraspecific genetic distances(2.42%and 8.66%).Three molecular analyses(NJ tree,ABGD,and GMYC)were performed to accurately identify and delineate species.Clustering errors occurred when the 12S rRNA sequences were delimited using the NJ tree method,and the delimitation results of ABGD and GMYC are consistent with the final species identification results.Our results demonstrate that DNA barcoding based on 12S rRNA and COI can be used as an effective tool for fish species identification,and 12S rRNA has good application prospects in the environmental DNA(eDNA)metabarcoding of marine fish.
基金Supported by the International Science Partnership Program of the Chinese Academy of Sciences(No.133137KYSB20200002)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA23050304)the Natural Science Foundation of Shandong Province(Nos.ZR2021MC151,ZR2021QD158)。
文摘The sea star Asterias amurensis is widely viewed as a severe“marine pest”because of its broad feeding habits.Over the past few decades,A.amurensis undergoes massive and sporadic population outbreaks worldwide,causing extensive economic and ecological losses to the local aquaculture industry and marine ecosystem.Understanding the genetic diversity and population structure of A.amurensis can provide vital information for resource management.By analyzing the polymorphism of the mitochondrial cytochrome C oxidase subunit I(COI)gene and ten simple sequence repeat(SSR)microsatellites markers,the genetic diversity and population structure of A.amurensis of four populations along the northern coast of China was uncovered.A total of 36 haplotypes were identified,and a main haplotype was found in four populations.The Qingdao(QD)population displayed the highest genetic diversity among all the populations.The AMOVA and pairwise F_(st)showed that there was small but statistically significant population differentiation among the four populations,especially between QD and Weihai(WH).Moreover,the principal component analysis(PCA)and admixture analysis showed that several individuals in Yantai(YT)and Dalian(DL)had little genetic association with other individuals.Overall,this study provided useful information of the genetic diversity and population structure of A.amurensis and will contribute to the resource management of A.amurensis in China.
基金Supported by National Natural Science Foundation of China:Mechanism of Protective Effects of Moxibustion on Regulating m TOR Signaling Pathway and Inhibiting Autophagy in Rats(No.81574084)Key Research and Development Program of Anhui Province:Mechanism and Clinical Application of Moxibustion in that Treatment of Chronic Heart Failure(No.202004j07020045)+1 种基金Open Fund Project of Key Laboratory of Xin’an Medical Education Ministry:Effect of Moxibustion on Myocardial Cell Energy Metabolism in Chronic Heart Failure Patients Based on Xin’an Physician’s Theory of Strengthening the Foundation and Cultivating the Original(No.2020xayx07)Key Natural Science Projects of Anhui Provincial Education Department:Mechanism of Moxibustion Against CHF Fibrosis Based on mi R-21/PTEN/m TOR Signaling Pathway-mediated Regulation of Autophagy in Myocardial Cells by circ PAN3(No.KJ2021A0570)。
文摘OBJECTIVE:To observe the effects of moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril on myocardial cells apoptosis index,the expression levels of apoptosis-related proteins cytochrome c(Cyt-C)and apoptosis-inducing factor(AIF)in chronic heart failure(CHF)rats.METHODS:Sixty-five rats were randomly divided into normal group(n=10)and model-I group(n=55).After modeling,CHF rats in model-I group were divided into model group,moxibustion group,benazepril group,moxibustion plus benazepril group(abbreviated as aibei group,the same below),10 rats in each group.Echocardiogram index was examined by echocardiography.Hemodynamic indices were measured by rat cardiac function meter.Serum B-type brain natriuretic peptide(BNP)was detected by enzymelinked immunosorbent assay.Myocardial cells apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining.Pathological changes of myocardial tissues were observed by hematoxylin and eosin staining.The expression levels of Cyt-C and AIF in myocardial tissues were detected by Western blot.RESULTS:Compared with normal group,ejection fraction and left ventricular diameter shortening rate in model-Ⅰgroup were significantly reduced,myocardial cells of rats in model group exhibited unclear transverse striations,cells swellings and vacuoles,cardiac functions were deteriorated,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly increased.Compared with model group,myocardial cells of rats in moxibustion group,benazepril group,and aibei group were dyed more evenly,muscle fibers were arranged relatively neatly,cardiac functions were improved,serum BNP level,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were significantly decreased.Compared with aibei group,cardiac functions were worsened,myocardial cells apoptosis index,and the expression levels of Cyt-C and AIF were increased.CONCLUSION:Moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)combined with benazepril could improve CHF better than moxibustion at bilateral Feishu(BL13)and Xinshu(BL15)or benazepril alone.The mechanisms might be that they can inhibit the expressions of Cyt-C and AIF,and inhibit the apoptosis of cardiomyocytes.