OBJECTIVEEndothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.

METHODSRAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.

RESULTSIncubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.

CONCLUSIONActivation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.

Animals ; Aorta ; cytology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Arecoline ; Carbachol ; Cell Cycle ; Endoplasmic Reticulum ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Homocysteine ; adverse effects ; Mitochondria ; metabolism ; Rats ; Receptors, Muscarinic ; metabolism

Vascular endothelium plays an important role in regulating vascular homeostasis. Over the past years, it has become clear that endothelial dysfunction is a key event of pathophysiological changes in the initiation and progression of injuries induced by extreme environmental factors. The present review summarizes current understanding of vascular endothelial dysfunction induced by hypoxia, cold and heat, and provides the information for prevention and treatment of environmental exposure injuries.
Endothelium, Vascular ; physiopathology ; Environment ; Humans ; Hypoxia ; physiopathology ; Temperature ; Vascular System Injuries

Objective To explore the effects of upper limb robot-assisted therapy on motor function and activities of daily living in con-valescent stroke patients. Methods From June to September, 2016, 12 chronic stroke patients at their first-ever stroke were enrolled and ran-domized into experimental group (n=6) and control group (n=6). Both groups received routine rehabilitation. Additional robot-assisted thera-py was provided to the experimental group, and additional repetitive movement training was provided to the control group, 20 minutes a day, five days a week for four weeks. Fugl-Meyer Assessment-Upper Extremities (FMA-UE), modified Ashworth Scale (MAS) and Func-tional Independent Measure (FIM) were used to assess the motor function of the upper limbs and hands, the muscular tension of shoulder and elbow, and activities of daily living (ADL) before and after treatment. Results After treatment, the scores of FMA-UE and FIM were bet-ter in both groups (Z>2.032, P<0.05), and no significant difference was found between two groups (t<0.723, P>0.05), however, the scores were a little bit higher in the experimental group than in the control group. After treatment, for the experimental group, the MAS scores of shoulder abduction/adduction and elbow flexion/extension improved (Z>2.121, P<0.05);for the control group, the MAS scores of shoulder abduction/adduction improved (Z>2.000, P<0.05), but the MAS scores of elbow flexion/extension were not significantly different (Z<1.890,P>0.05). There was no significant difference in the MAS scores of shoulder abduction/adduction and elbow flexion/extension between two group (Z<1.734, P>0.05). The moving trail recorded by the computer, gradually became a regular pattern from the mass, saying the motor control ability became better. Conclusion Upper limb robot-assisted therapy can promote the recovery of the motor function of upper limbs and ADL in convalescent stroke patient, similar to the repetitive movement training.

Objective To investigate the change of serum Tg levels of DTC patients with positive stimulated Tg (Tg ≥ 10.00 μg/L),negative 131I-diagnostic whole body scan(Dx-WBS) and no distant metastasis 6 months after initial 131I therapy.Methods Fifty-six DTC patients (20 males,36 females,average age 43.11 (21-70) y) with intermediate or low risk of recurrence according to American Thyroid Association (ATA) guideline were enrolled into the retrospective study.All patients were grouped according to stimulated Tg level after initial 131I therapy:group with positive Tg (Tg+ group,n =19) and group with negative Tg (Tgˉ group,n=37).Changes of suppressed Tg at 1 year and 2.5 years (Tg1ysup and Tg2.5ysup) after initial therapy were compared between the two groups.Serum TSH level,TgAb level,neck ultrasound and chest CT results were also evaluated.The two-sample t test and x2 test were used for statistical analysis with SPSS 17.0.Results Stimulated Tg and Tglysup levels in Tg+ group were remarkably higher than those in Tgˉ group:(24.27±4.10) μg/L vs (2.73±3.01) μg/L,t=7.191,P＜0.05(6 months after initial 131I therapy) ; (2.21±0.55) vs (0.48±0.10) μg/L,t=3.102,P＜0.05(1 year after initial 131I therapy),respectively.In Tg+ group,suppressed Tg level decreased with time in 68.4％ (13/19) of patients,of whom the Tg2.5ysup level was much lower than Tglysup level ((0.53±0.15) μg/L vs (1.38±0.50) μg/L).Tg2.5sup level in Tg+ group became comparable to that in Tgˉ group ((1.44±0.52) μg/L vs (0.38±0.07) μg/L; t =2.001,P＞0.05).In each group,one case of recurrence with suppressed Tg of 1.4 μg/L and 0.1 μg/L respectively,was observed using neck ultrasound after 2 years of follow-up.Conclusions Serum Tg levels decreased with time for Tg+/131I-Dx-WBS-DTC patients with intermediate or low risk of recurrence.It might not be necessary to follow up these patients with Tg and 131 I-DxWBS after 6 months of initial 131I therapy.

The incidence of deacclimatization to high altitude syndrome (DAHAS) prevailed up to 80% in highland troops, and 100% in manual workers, and severe DAHAS could significantly affects patients' health, work and life. So it is imperative to develop effective prevention and treatment measures for DAHAS. The present review analyzes effective prophylactic and therapeutic measures against DAHAS, implemented at our hospital.
Acclimatization ; Altitude ; Altitude Sickness ; prevention & control ; therapy ; Humans

OBJECTIVEHigh altitue pulmonary edema (HAPE) impacts seriously people's health at high altitude. Screening of susceptibility genes for HAPE will be used for the evaluation and protection of susceptible people.

METHODSWe performed a genome-wide association study (GWAS) using Affymetrix SNP array 6.0 in 23 HAPE patients and 17 healthy controls. GO and Pathway analysis softwares were used to analyze and draw gene network.

RESULTSThirty-nine SNPs were found to be significantly different between case and control groups (P < 10(-4)). GO and Pathway analysis of 27 genes around the 39 SNPs indicated that these genes mainly participate in the regulating of cell proliferation, regulation of nitrogen compound metabolic process and G-protein coupled receptor protein signaling pathway and so on.

CONCLUSIONIt suggests that these SNPs and genes found in this study may be associated with the susceptibility of HAPE.

Adult ; Altitude Sickness ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Hypertension, Pulmonary ; genetics ; Polymorphism, Single Nucleotide ; Young Adult

The primary object of this fundamental research was to survey the synergistic cardiovascular effects of iptakalim, a novel ATP-sensitive potassium channel (K(ATP)) opener, and clinical first-line antihypertensive drugs, such as calcium antagonists, thiazide diuretics and β receptor blockers by a 2 x 2 factorial-design experiment. It would provide a theoretical basis for the development of new combined antihypertensive therapy program after iptakalim is applied to the clinic. Amlodipine besylate, hydrochlorothiazide and propranolol were chosen as clinical first-line antihypertensive drugs. Blood pressure, heart rate (HR) and cardiac functions were observed in anesthetized normal rats by an eight-channel physiological recorder. The results showed that iptakalim monotherapy in a low dose could produce significant antihypertensive effect. There was no interaction between iptakalim and amlodipine on the maximal changes of systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial blood pressure (MABP), the left ventricular systolic pressure (LVSP), and the left ventricular end-diastolic pressure (LVEDP) (P > 0.05). However, the effects of combination iptakalim/amlodipine on the maximal changes of SBP, DBP, MABP, LVSP and LVEDP were more obvious than those of iptakalim or amlodipine monotherapy. And there was strong positive interaction between iptakalim and amlodipine on the maximal changes of HR (P>0.05). According to the maximal changes of DBP, MABP, LVSP and LVEDP (P < 0.05) of combination iptakalim with hydrochlorothiazide, there was strong positive interaction between them. But there was no interaction between iptakalim and hydrochlorothiazide on the maximal drop of SBP and HR (P > 0.05). According to the maximal drops of DBP, MABP of combination iptakalim with propranolol, there was strong positive interaction between them (P < 0.05). But there was no interaction between iptakalim and propranolol on the maximal changes of SBP, LVSP, LVEDP and HR (P > 0.05). In conclusion, it was the first time to study the effects of amlodipine, hydrochlorothiazide or propranolol, which had different mechanisms of action from iptakalim, on cardiovascular effects of iptakalim in anesthetized normal rats. This study proved that the combination of iptakalim with hydrochlorothiazide or propranolol respectively had significant synergism on lowering blood pressure, while the combination of iptakalim/amlodipine had additive action on lowering blood pressure. Meanwhile the antihypertensive effect was explicit, stable and long-lasting. Iptakalim thus appears suitable for the clinical treatment of hypertensive people who need two or more kinds of antihypertensive agents.
Amlodipine ; pharmacology ; Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; drug effects ; Drug Synergism ; Heart Rate ; Hydrochlorothiazide ; pharmacology ; Hypertension ; Propranolol ; pharmacology ; Propylamines ; pharmacology ; Rats

OBJECTIVETo investigate the effects of non-neuronal muscarinic receptors (NNMR) stimulation on atherosclerosis and endothelial cells activation.

METHODSAtherosclerosis model was established in ApoE-/- mice by a high fat diet for 7 weeks. During the experimental periods, animals were received a low (7 mg/kg/d) or a high (21 mg/kg/d) dose of arecoline by gavage. At the termination of the treatments, serum total cholesterol and NO levels were measured, and the aorta morphology was analyzed by hematoxylin and eosin staining. The gene expression of monocyte chemoattractant protein-1 (MCP-1) and adhesion molecules in the thoracic aortas was determined by RT-PCR, and the MCP-1 protein expression and NF-κB activity were detected by Western blot analysis. NO production, MCP-1 secretion in cultured rat aortic endothelial cells (RAECs), and monocyte-endothelium adhesion assay were also performed after arecoline treatments.

RESULTSArecoline efficiently decreased atherosclerotic plaque areas, increased serum nitric oxide (NO) content, suppressed the mRNA and protein expression of MCP-1, and modulated the IκB-α degradation and P65 phosphorylation in the aortae of ApoE-/- mice. Furthermore, arecoline promoted NO production and suppressed MCP-1 secretion in cultured RAECs after ox-LDL exposure, and either atropine or NG-nitro-L-arginine methylester could abrogate these effects. Arecoline also significantly inhibited the adherence of U937 monocytes to the ox-LDL injured human umbilical vein endothelial cells, which could be abolished by atropine.

CONCLUSIONOur results indicate that arecoline attenuates the progression of atherosclerosis and inhibits endothelial cells activation and adherence by stimulating endothelial NNMR. These effects, at least in part, are due to its modulation on NF-κB activity.

Animals ; Aorta ; cytology ; Apolipoproteins E ; Arecoline ; pharmacology ; Atherosclerosis ; physiopathology ; prevention & control ; Cell Adhesion Molecules ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Disease Progression ; Endothelial Cells ; cytology ; drug effects ; Endothelium, Vascular ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; I-kappa B Proteins ; metabolism ; Lipoproteins, LDL ; Mice ; Mice, Knockout ; Monocytes ; cytology ; NF-KappaB Inhibitor alpha ; Nitric Oxide ; blood ; Nitroarginine ; pharmacology ; Rats ; Receptors, Muscarinic ; physiology ; Transcription Factor RelA ; metabolism

AIMTo compare the differences of pharmacological characteristics of the endothelial target for acetylcholine (ETA) between rat aorta and tail artery.

METHODSDifferences in the endothelium-dependent relaxation induced by acetylcholine (ACh: 10(-8) - 10(-4) mol/L) were studied using isolated rat tail artery helical strips and aortic rings, so that the pharmacological characteristics of ETA in small artery can be observed.

RESULTSACh-induced endothelium-dependent relaxation was observed both in rat tail artery strips and in aortic rings precontracted with potassium chloride (60 mmol/L) in a concentration-dependent manner. In tail artery this effect was partially blocked by L-N(omega)-Nitro-arginine methyl ester (L-NAME: 10(-4) mol/L) or methylene blue (MB: 10(-5) mol/L), together with indomethacin (Indo: 10(-4) mol/L), but in aorta it was completely blocked by L-NAME or MB.

CONCLUSIONIt is different of the pharmacological characteristics of ETA between big artery and small artery. A non-NO and non-PGI2 relaxing factor, together with nitric oxide (NO) and prostacyclin (PGI2), mediates endothelium-dependent vasorelaxation induced by ACh in small artery, but NO may be the principal endothelial vasodilator substance in big artery.

Acetylcholine ; pharmacology ; Animals ; Aorta ; drug effects ; Arteries ; drug effects ; Endothelium, Vascular ; drug effects ; physiology ; In Vitro Techniques ; Male ; Rats ; Rats, Wistar ; Vasodilation ; drug effects

AIMIn order to evaluate the regulatory effects of nucleotides and adenosine on ATP-sensitive potassium channel (K(ATP)) in artery smooth muscles, the effects of them on vascular relaxation induced by K(ATP) opener pinacidil(Pin) were investigated.

METHODSThe isolated endothelium- denuded aorta rings were preincubated with nucleotides or nucleotides and glibenclamide (Gli) for 10 min, the vascular relaxation induced by Pin in aorta precontracted with 20 mmol x L(-1) KCl was observed.

RESULTSAfter the isolated endothelium-denuded aorta rings were preincubated with ATP, ADP, UDP, GTP and adenosine (Ade) 100 micromol x L(-1) respectively, the vascular relaxation induced by Pin was changed as following: (1) ATP could inhibit the K(ATP) activation by Pin and enhance the blockade of K(ATP) by Gli. (2) ADP could inhibit the K(ATP) activation by Pin and attenuate the blockade of K(ATP) by Gli. (3) The regulatory effect of Ade on K(ATP) was similar with that of ADP. (4) UDP could enhance the K(ATP) activation by Pin and attenuate the blockade of K(ATP) by Gli. (5) GTP could enhance the K(ATP) activation by Pin, but had no effects on the blockade of K(ATP) by Gli.

CONCLUSIONNucleotides and adenosine, related to energy metabolism, could modulate the functions of K(ATP) in vascular smooth muscle. But their pharmacological characteristics were different.

Animals ; Aorta ; cytology ; drug effects ; Glyburide ; pharmacology ; In Vitro Techniques ; KATP Channels ; metabolism ; Male ; Muscle, Smooth, Vascular ; drug effects ; Nucleotides ; pharmacology ; Pinacidil ; pharmacology ; Rats ; Rats, Wistar ; Vasodilator Agents ; pharmacology