Diabetes mellitus is a common disease of serious harmful to human health,but its pathogenesis is not entirely clear.Mitochondria are the important organelles to generate energy in eukaryocytes,and play a pivotal role in the regulation of reactive oxygen species generation,intracellular calcium homeostasis,and apoptosis signal transduction.The possible causes of mitochondrial dysfunction include oxidative stress,Ca2+ disturbances,reduction of mitochondrial biosynthesis,opening of mitochondrial permeability transition pore,mitochondrial DNA mutations,and etc..Many studies demonstrate that mitochondrial dysfunction is associated with β-cell dysfunction of type 1 diabetes mellitus and insulin resistance of type 2 diabetes mellitus.Moreover,mitochondrial dysfunction plays important roles in the development of diabetic cardiomyopathy.This present article reviewed the current status of studies on the relationship of mitochondrial dysfunction and diabetes mellitus and diabetic cardiomyopathy.It is very important to understand and study mitochondrial dysfunction and its important roles in diabetes mellitus and diabetic cardiomyopathy in order to clarify the pathogenesis of diabetes mellitus and explore new approaches of prevention and treatment for diabetes mellitus.

Objective To investigate the effects of cold exposure with different time courses on cardiovascular function and skeletal muscle contraction in Sprague-Dawley (SD) rats.Methods The male SD rats were randomly divided into control,12-h cold,24-h cold,and 48-h cold groups.The rats in all cold group were maintained in 4 ℃ environment for indicated time and allowed free access to standard rat chow and water ad libitum.After the animal model was established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Lanendorff heart persusion was used to detect some parameters of cardiac function,such as coronary flow (CF),mean left ventricular systolic pressure (mLVSP),mean left ventricular diastolic pressure (mLVDP) and maximum ascending and declining rate of left ventricular pressure (± dp/dt max) ; and then the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes of above cardiac function indexes were recorded again.Organ bath method was employed to detect contraction response to potassium chloride (KCl) or phenylephrine (PE) and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings.The twitch tension,titanic tension,and fatigue test of soleus and EDL were examined by electrical stimulation to reflect contractive function of skeletal muscle.Results The coronary flow (CF) were significantly reduced in 24-h and 48-h cold groups compared to control group [(9.0 ± 1.7) ml/g and (9.8 ± 1.3) ml/g vs (11.6 ± 1.2) ml/g,P ＜ 0.05] ; 48-h cold exposure also showed significantly increased mLVSP (P ＜0.01) and mLVP (P ＜ 0.05) of rats.After ischemia-reperfusion,the heart re-beat time was shorter in all cold exposure groups (all P ＜ 0.01),mLVP and mLVDP were higher in 24-h and 48-h cold group than control group (all P ＜ 0.01).The contraction of aortic rings responses to 60 mmol/L KCl [(0.63 ±0.13) g and (0.79 ±0.11)g vs (1.28±0.17)g,P ＜0.01] and 1 μmol/L PE [(0.62 ±0.18)g and (0.68 ±0.13)g vs (1.25 ± 0.18)g,P ＜ 0.01] were decreased while the endothelium-dependent relaxation responses to low concentration ACh (3 × 10-8 ～ 3 × 10-7) were increased in 24-h and 48-h cold groups relative to control group (all P ＜ 0.05).The 48-h cold also reduced titanic tension of SOL and EDL in comparison with control [(4.14 ± 0.62) N/cm2 vs (5.50 ± 0.75) N/cm2,P ＜ 0.05 and (3.00 ± 0.57) N/ cm2 vs (4.70 ± 0.85) N/cm2,P ＜ 0.01].Conclusions 24 ～ 48 h cold exposure could reduce CF but enhance the tolerance of heart from rats to ischemia reperfusion injury and also decrease the contraction of aortic rings response to KCl or PE but increase the sensitive responses to ACh of aortic rings from rats.The 48-h cold could decrease the contraction of skeletal muscle elicited by electrical stimulation.

Objective To investigate the effects of fasting with different time courses on the contraction of skeletal muscle and cardiovascular function in Sprague-Dawley (SD) rats.Methods The healthy and male SD rats were randomly divided into (1) control,(2) 12 h fasting,(3) 24 h fasting,and (4) 48 h fasting groups.After animal models of fasting with different time courses were established,the heart,thoracic aorta,soleus,and extensor digitorum longus (EDL) were rapidly isolated from rats under anesthesia and then fixed into the respective organ perfusion system that was connected to physiological recorders and data acquisition system.Langendorff heart perfusion was used to detect some parameters of cardiac function,including left ventricular systolic pressure (LVSP),left ventricular end-diastolic pressure (LVEDP),and maximum ascending and declining rate of left ventricular pressure (± dP/dt max).Then,the heart was suffered from 30 min ischemia followed by 60 min reperfusion,the changes in above cardiac function parameters were recorded again.The contraction response to potassium chloride or phenylephrine and endothelium-dependent relation responses to acetylcholine (ACh) of aortic rings were measured with organ bath.The changes of twitch tension and titanic tension of soleus and EDL were examined to reflect contractive function of skeletal muscle.Results The + dP/dt max was significantly reduced in 24 h and 48 h fasting groups compared to the control (P ＜ 0.01).After ischemia-reperfusion,the heart re-beat time was significantly longer [(24.3 ± 7.0)min vs (14.0 ± 2.9)min,P ＜0.05],both mLVDP and mLVP were significantly higher in 48 h and 24 h fasting groups than control group (P ＜ 0.05),and those changes did not normalized until re-perfusion 60 min.The 48 h fasting increased significantly endothelium-dependent relaxation of aortic rings response to low concentration acetylcholine (P ＜ 0.05),but did not affect the maximum relaxation of aortic rings response to high concentration acetylcholine and contraction of aortic rings responses to potassium chloride and phenylephrine.The 48 h fasting also enhanced significantly titanic tension of soleus compared to the control [(7.01 ± 1.22) N/cm2 vs (5.5 ± 0.75) N/cm2 for48 h-Fasting vs Control,P ＜0.05].Conclusions The 24 ～48 h fasting may induce inhibition of cardiac function,while fasting 48 h can increase the sensitivity of aortic rings responses to ACh and contraction function of soleus in healthy and male rats.

[ ABSTRACT] AIM:To observe the effect of endogenous nitric oxide synthase ( NOS) inhibitor asymmetric dimeth-ylarginine ( ADMA ) and its signaling pathways on NO levels and skeletal muscle contractility in 4-week running rats. METHODS:The 4 weeks running rat model was established.The twitch tension, tetanic tension and the fatigue test of sole-us muscle induced by electrical stimulation ex vivo were detected.The ATP content, mitochondrial DNA levels and the mR-NA expression of peroxisome proliferator-activated receptor γcoactivator-1α(PGC-1α), nuclear respiratory factor (NRF) were measured to reflect the mitochondrial function and biosynthesis in the skeletal muscle.Serum ADMA concentration was detected by high performance liquid chromatography.The endogenous ADMA enzymes PRMT1 and 2 subtypes of ADMA me-tabolism enzyme DDAH, 3 subtypes of NOS protein expression in the skeletal muscle were determined by Western blot.NOS activity and nitric oxide ( NO) content were analyzed by colorimetric method.RESULTS: Compared with normal control group, the twitch tension, tetanic tension and the anti-fatigue capability of soleus muscle in running group were significantly enhanced, ATP content, mitochondrial DNA content and the mRNA expression of PGC-1αand NRF were significantly in-creased (P<0.01).In addition, the protein expression of constitute type NOS (cNOS) and NOS activity were significantly increased (P<0.01), but the increase in NO content was relatively smaller in soleus muscle in exercise group (P<0.05). Moreover, serum ADMA concentration in running group was increased, while the DDAH2 expression in skeletal muscle was decreased.CONCLUSION:Short-term endurance exercise enhances the twitch tension, tetanic tension and fatigue resist-
ance of soleus muscle.The mechanism may be that increased cNOS expression feedbacks to increase ADMA concentration, thus maintaining the increase in NO synthesis at a relatively low level, and resulting in promoting skeletal muscle mitochon-dria biosynthesis and mitochondrial function.

AIM:To investigate the influence of calorie restriction ( CR) on contractility and mitochondrial bi-osynthesis in different types of rat skeletal muscles .METHODS:CR rat model was set up by feeding 60%normal food in-take of control rat every day for 8 weeks.Soleus (SOL) and extensor digitorum longus (EDL) were isolated under anesthe-sia.The twitch tension, titanic tension and fatigue resistance of SOL and EDL in response to electrical stimulation were measured to reflect the contractile function of the muscles .The copy number ratio of mitochondrial gene cytochrome C oxi-dase subunit I ( COX I) to nuclear gene β-actin was determined to evaluate the mitochondrial biosynthesis .ATP content was measured to mirror mitochondrial function .RESULTS:Compared with control group , CR for 8 weeks significantly in-creased twitch tension and titanic tension of both SOL and EDL , but only improved fatigue resistance in SOL .Markedly in-crease in ATP content in both skeletal muscles by CR intervention was observed , especially in SOL .Although CR activated AMP-activated protein kinase (AMPK) in both 2 muscles, up-regulation of mitochondrial biosynthesis and transcription of mitochondrial regulatory genes peroxisome proliferator-activated receptor γcoactivator 1α( PGC-1α) and nuclear respirato-ry factor ( NRF) was only observed in SOL .CONCLUSION:CR for 8 weeks enhanced the contractility of both rat SOL and EDL in response to electrical stimulation , especially in SOL composed of slow-twitch fibers.The mechanisms may be related to the activation of AMPK and the promotion of mitochondrial biosynthesis in SOL .

Aim This study was aimed to explore the influence and mechanism of the long-term exercise on skeletal muscle contraction and mitochondrial biosyn-thesis in different muscle fibers.Methods Soleus (SOL)and extensor digitorum longus (EDL)were i-solated from SD male rats with platform running train-ing for eight weeks.The changes of contractility under different electrical stimulation were observed, mito-chondrial biosynthesis,including ATP content,mito-chondrial DNA,the gene expression of PGC-1αand NRF were also detected.Results Long-term endur-ance exercise can improve twitch tension and titanic tension of SOL and EDL ,but only enhanced the fa-tigue resistance in SOL.ATP contents were significant-ly increased in the two types of muscles,but mtDNA content,PGC-1αexpression and NRF translation were only obviously enhanced in SOL,in accompanied with an increase in p-AMPK/AMPK protein ratio.Conclu-sion Long-term endurance exercise increased skeletal muscle contractility and improved the anti-fatigue abili-ty in SOL,which may be associated with increase in mitochondrial biosynthesis via activated AMPK-PGC-1αaxis.

Objective To investigate the effects of calorie restriction (CR) for 4 weeks on twitch tension,titanic tension,and fatigue contraction induced by electrical stimulation in different kind skeletal muscles from rats and explore the possible mechanisms.Methods The rat model of CR was established by a limitation of 40％ calorie intake of control rats for 4 weeks,and then oral glucose tolerance test (OGTT) was performed.The soleus (SOL) and extensor digitorum longus (EDL) were isolated under anesthetization to detect twitch tension,titanic tension,and fatigue contraction induced by electrical stimulation.Adenosine triphosphate (ATP) content was measured by fluorescent enzymatic methods to reflect mitochondrial function.The ratio of mitochondrial gene COX Ⅰ and nuclear gene β-actin copy number was analyzed to evaluate mitochondrial biogenesis.Furthermore,the transcriptions of peroxisome proliferatoractivated receptor γ coactivator-1 (PGC-1α) and nuclear respiratory factor 1 (NRF1) genes,expressions of phosphorylated adenosine 5'-monophosphate-activated protein kinase (AMPK) and nitric oxide synthase (NOS) proteins,and nitric oxide (NO) content were determined in skeletal muscle.Results The blood glucose level at 30 min and area under the curve of blood glucose levels at various time points during OGTT were significantly decreased in the CR group compared to the control group.The twitch tension [(2.5 ± 0.15)N/cm2 vs (1.24±0.12)N/cm2,(2.66 ±0.21)N/cm2 vs (1.69 ±0.17)N/cm2,P ＜ 0.05],titanic tension [(10.43 ± 0.36) N/cm2 vs (8.06 ± 0.19) N/cm2,(11.35 ± 1.02) N/cm2 vs (8.12 ± 0.23) N/cm2,P ＜ 0.05],and fatigue contraction force in SOL and EDL from CR rats were significantly increased in association with increases of ATP content [(34.82 ±4.31)) mnol/mg protein vs (15.32 ± 1.94) nmol/mg protein,(30.82 ± 2.15) nmol/mg protein vs (12.32 ± 0.97) nmol/mg protein,P ＜ 0.05] and mitochondrial biogenesis (2.75 ± 0.20 vs 1.52 ± 0.06,1.32 ± 0.10 vs 0.84 ± 0.11,P ＜ 0.05) compared to control rats.CR for 4 weeks upregulated the transcriptions of PGC-1α and NRF genes as well as the phosphorylation of AMPK protein in SOL but not in EDL.Furthermore,CR also enhanced NOS expression and NO content in both skeletal muscles.Conclusions CR for 4 weeks can strengthen the contractile function of SOL and DL from rats,and the underlying mechanisms might be related to the upregulation of PGC-1α transcription and AMPK activation,resulting in the enhances of mitochondrial biogenesis and mitochondrial function in skeletal muscles.

Integrase plays a critical role in the recombination of viral DNA into the host genome. Therefore, over the past decade, it has been a hot target of drug design in the fight against type 1 human immunodeficiency virus (HIV-1). Bovine immunodeficiency virus (BIV) integrase has the same function as HIV-1 integrase. We have determined crystal structures of the BIV integrase catalytic core domain (CCD) in two different crystal forms at a resolution of 2.45 Å and 2.2 Å, respectively. In crystal form I, BIV integrase CCD forms a back-to-back dimer, in which the two active sites are on opposite sides. This has also been seen in many of the CCD structures of HIV-1 integrase that were determined previously. However, in crystal form II, BIV integrase CCD forms a novel face-to-face dimer in which the two active sites are close to each other. Strikingly, the distance separating the two active sites is approximately 20 Å, a distance that perfectly matches a 5-base pair interval. Based on these data, we propose a model for the interaction of integrase with its target DNA, which is also supported by many published biochemical data. Our results provide important clues for designing new inhibitors against HIV-1.
Animals ; Catalytic Domain ; genetics ; Cattle ; DNA ; genetics ; DNA, Viral ; HIV-1 ; genetics ; metabolism ; Humans ; Immunodeficiency Virus, Bovine ; enzymology ; genetics ; Integrases ; chemistry ; genetics ; metabolism

Edong Healthcare Group is cited as an example to summarize its practice of centralized purchasing of supplies in the recent two years.In addition to a success in its process and mechanism, the authors analyzed some challenges for the purpose of furthering the group-based reform of public hospitals.

OBJECTIVES: Our previous study has verified that high level of SET and MYND domain-containing protein 2 (SMYD2) plays an important role in acquiring aggressive ability for liver cancer cells in hepatocellular carcinoma. MiR-200b as a tumor suppressor gene involves in a variety of cancers. This study aims to investigate the correlation between miR-200b and SMYD2 in hepatocellular carcinoma and the underlying mechanism. METHODS: Firstly, the levels of SMYD2 and miR-200b in hepatocellular carcinoma tissues and matched adjacent non-tumor liver tissues were tested with real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Secondly, we evaluated the interaction between miR-200b and SMYD2 using dual-luciferase reporter assay. Thirdly, we elucidated the effect of miR-200b on SMYD2 and its downstream targets p53/CyclinE1. Finally, we silenced SMYD2 in hepatocellular carcinoma cell lines to investigate its effect on tumor proliferation and cell cycle progression, and further confirmed the correlation among SMYD2 and p53/CyclinE1. RESULTS: Compared with the matched adjacent non-tumor liver tissues, miR-200b was obviously decreased, and SMYD2 was significantly increased in hepatocellular carcinoma (both P<0.05). Spearman's rank correlation revealed that miR-200b expression was negatively correlated with SMYD2 (P<0.01). Computer algorithm and dual-luciferase reporter assay revealed that miR-200b directly targeted and suppressed SMYD2 in HEK 293T cells. The down-regulated miR-200b expression promoted hepatoma cell proliferation (P<0.05) and increased SMYD2 expression(P<0.01), while the up-regulated expression of miR-200b had an opposite effect. The knockdown of SMYD2 suppressed the proliferation of MHCC-97L cells (P<0.01), down-regulated CyclinE1, and up-regulated p53 expression (both P<0.05). CONCLUSIONS MiR-200b is involved in hepatocellular carcinoma progression via targeting SMYD2 and regulating SMYD2/p53/CyclinE1 signaling pathway and may be used as a potential target for hepatocellular carcinoma treatment.
Humans ; Carcinoma, Hepatocellular/pathology* ; Tumor Suppressor Protein p53/metabolism* ; MicroRNAs/metabolism* ; Cell Line, Tumor ; Signal Transduction ; Liver Neoplasms/pathology* ; Cell Proliferation/genetics* ; Histone-Lysine N-Methyltransferase/metabolism*