Serum levels of vasoactive factors in HIV-infected pre-eclamptic women on HAART.

In South Africa, pre-eclampsia (PE) and human immunodeficiency virus (HIV) infection are major causes of pregnancy-related deaths. This study aimed to measure serum levels of endothelin-1 (ET-1), endothelial nitric oxide synthase (eNOS), soluble fms-like tyrosine kinase 1 (sFlt-1), soluble endoglin (sEng) and placental growth factor (PlGF) in HIV-infected highly active antiretroviral therapy (HAART)-treated and HIV-uninfected PE and normotensive women to ascertain if HIV/HAART alters their concentrations. Mean sFlt-1 levels were significantly up-regulated in the PE (HIV-uninfected 4.39 ± 1.29; HIV-infected 5.10 ± 1.10 ng/ml) compared to normotensive women (HIV-uninfected 2.59 ± 0.83; HIV-infected 2.20 ± 0.85 ng/ml). Mean PlGF levels were significantly lower in HIV-uninfected PE vs. HIV-infected normotensive women (29.69 ± 4.47 pg/ml vs. 32.86 ± 6.46 pg/ml; p = .002). In conclusion, PE women with HIV exhibited significantly low serum PlGF, ET-1 and eNOS levels. Infection with HIV may have further increased the sFlt-1 levels.IMPACT STATEMENTWhat is already known on this subject? In PE, the numerous identified local and circulating bioactive factors differed in concentrations when compared to normal pregnancy.What do the results of this study add? PE women with HIV exhibited significantly low serum PlGF, ET-1 and eNOS levels as well as increased levels of sFlt-1.What are the implications of these findings for clinical practice and/or further research? Understanding the link between PE, HIV and HAART during pregnancy will improve prognosis, management and treatment strategies for women clinically.

Identification and quantification of the quality markers and anti-migraine active components in Chuanxiong Rhizoma and Cyperi Rhizoma herbal pair based on chemometric analysis between chemical constituents and pharmacological effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Chuanxiong Rhizoma and Cyperi Rhizoma (CRCR), an ancient and classic herbal pair, has been used in herbal medicines for treating migraine, but its effective components are not clear. AIM OF THE STUDY: The present study aimed to identify and quantify the quality markers and anti-migraine active components in CRCR based on chemometric analysis between chemical constituents and pharmacological effects. MATERIALS AND METHODS: The HPLC fingerprints of eight batches of CRCR samples were obtained, and their characteristic common peaks were identified by HPLC-ESI-Q-TOF-MS/MS. The therapeutic effects of eight batches of CRCR samples on nitroglycerin-induced migraine rats were evaluated by migraine-related neurotransmitters and neuropeptides. Similarity analysis, hierarchical cluster analysis and principal component analysis were applied to screen the quality markers. Artificial neural network and partial least squares regression models were used to screen the anti-migraine compounds by correlating the chemical constituents in HPLC fingerprints and pharmacological indicators. RESULTS: Eighteen characteristic common peaks were found in the HPLC fingerprints, including eleven known compounds and seven unknown compounds. Ferulic acid (FA), senkyunolide I (SI), senkyunolide A (SA), 3-n-butylphthalide (NBP), Z-ligustilide (LIG), Z-3-butylidenephthalide (BDPH), nookatone (NKT), levistilide A (LA), α-cyperone (CYP) and other five unknown compounds (P1, P2, P7, P8 and P9) were identified as quality markers. SA, NBP, LIG, NKT, CYP and other three unknown compounds (P1, P4 and P9) can be considered as anti-migraine prototype compounds. The quality markers and anti-migraine active components were further quantified in CRCR extract, rat serum and cerebral cortex by UPLC-MS/MS, which gives a clue to track the dynamic changes of the contents of the main constituents. CONCLUSIONS: Our study explored the anti-migraine material basis, and could lay a foundation for the improvement of the quality control of CRCR in practice.

Effect of an aqueous extract of Averrhoa carambola L. on endothelial function in rats with ventricular remodelling.

Ventricular remodelling leads to cardiomyocyte hypertrophy, myocardial fibrosis, endothelial vasoactive substance changes and endothelial dysfunction. Our purpose was to research the effect of an aqueous extract of Averrhoa carambola L. (AEA) on endothelial function in rats with ventricular remodelling induced by isoprenaline. Rats were subjected to injection of isoprenaline and administration of various drugs. Vasoactive substances were measured, and the ventricular remodelling index was detected by the weighing method. Immunohistochemical analysis, pathological examination, Western blot and Masson's trichrome staining were performed. After AEA administration, the levels of transforming growth factor-ß (TGF-ß), angiotensin II (AngII), inducible NO synthase (iNOS), endothelin-converting enzyme (ECE), and endothelin 1 (ET-1); the ventricular remodelling index; and the collagen volume fraction were decreased, while the levels of total NO synthase (tNOS) and endothelial NO synthase (eNOS) were increased. The pathological examination results showed that apoptosis, fibrosis, necrosis and inflammatory infiltration of myocardial tissue were attenuated by AEA treatment. AEA might alleviate ventricular remodelling in rats by maintaining the balance of vasoactive substances and the function of the vascular endothelium.

Marcadores de estrés oxidativo en pacientes con esclerosis múltiple durante un brote y su remisión / Oxidative stress markers in patients with multiple sclerosis during an outbreak and its remission / Marcadores de estresse oxidativo em pacientes com esclerose múltipla durante um surto e sua remissão

La esclerosis múltiple remitente-recurrente (EM-RR) es una enfermedad desmielinizante del sistema nervioso central. A fin de entender la asociación del estrés oxidativo a nivel periférico con la recaída de la enfermedad se determinaron los niveles de marcadores de estrés oxidativo en plasma de pacientes en la recaída o brote y una semana después de la misma. Se analizaron muestras de 60 personas (20 pacientes con recaída, 20 pacientes sin recaída y 20 controles sanos). Se cuantificaron mediante métodos espectrofotométricos las actividades enzimáticas de óxido nítrico sintasa (ONS), glutatión peroxidasa (GPx), los niveles de lipoperóxidos y nitritos-nitratos y la fluidez de membrana. En el brote de la enfermedad aumentan significativamente los niveles de las actividades enzimáticas de ONS y GPx y los niveles de nitritos-nitratos y lipoperóxidos (p<0,01 en todos los casos), al ser comparados con los de individuos sanos. Dichos parámetros disminuyeron significativamente una semana después de iniciado el brote. Además, los parámetros evaluados se mantuvieron elevados en pacientes que no experimentaron un brote de la enfermedad cuando se los comparó con individuos sanos. La fluidez de membrana en los pacientes con y sin brote fue similar a la de los controles. En conclusión, el estrés oxidativo es un componente importante en los pacientes con esclerosis múltiple.

Recurrent-remitting multiple sclerosis (RR-MS) is a demyelinating disease of the central nervous system. In order to understand the association of oxidative stress at the peripheral level with the relapse of the disease, the levels of oxidative stress markers in plasma of patients in the relapse or outbreak and one week after relapse were determined. Samples of 60 subjects were analyzed (20 patients in relapse, 20 patients without relapse, and 20 healthy controls). The enzymatic activities of nitric oxide synthase (NOS), glutathione peroxidase (GPx), lipoperoxides and nitrite-nitrate levels and membrane fluidity were quantified by spectrophotometric methods. In relapse, the levels of enzymatic activities of NOS and GPx, and the levels of lipoperoxides and nitrites-nitrates were significantly increased (p<0.01, in all cases), compared with healthy individuals. These parameters decreased significantly 1 week after the start of the outbreak. In addition, the parameters evaluated remained high in patients who did not experience an outbreak of the disease compared to healthy subjects. The membrane fluidity in the patients with and without outbreak was similar to that of the controls. In conclusion, oxidative stress is an important component in patients with multiple sclerosis.

A esclerose múltipla recorrente-remitente (EM-RR) é uma doença desmielinizante do sistema nervoso central. Para compreender a associação do estresse oxidativo a nível periférico com a recaída da doença foram determinados os níveis de marcadores de estresse oxidativo em plasma de doentes na recaída ou surto e uma semana após a recaída. Foram analisadas a amostras de 60 pessoas (20 pacientes com recaída, 20 pacientes sem recaída e 20 controles saudáveis). As atividades enzimáticas de óxido nítrico sintase (ONS), glutationa peroxidase (GPX), os níveis de lipoperóxidos e nitritos-nitratos e a fluidez de membrana foram quantificadas por métodos espectrofotométricos. No surto da doença aumentam em forma significativa os níveis da atividade enzimática de ONS e GPX, e os níveis de nitritos-nitratos e lipoperóxidos (p<0,01 em todos os casos), em comparação com os indivíduos saudáveis. Esses parâmetros diminuíram significativamente uma semana após o início do surto. Além disso, os parâmetros avaliados permaneceram elevados em pacientes que não experimentaram um surto da doença quando comparados com indivíduos saudáveis. A fluência de membrana nos pacientes com e sem surto foi semelhante à dos controles. Em conclusão, o estresse oxidativo é um componente importante nos pacientes com esclerose múltipla.

Serum levels of total antioxidant status, nitric oxide and nitric oxide synthase in minor recurrent aphthous stomatitis patients.

BACKGROUND: Recurrent aphthous stomatitis (RAS) is one of the most common inflammatory ulcerative conditions of oral cavity with uncertain etiology. Several studies have reported that oxidative stress may be associated with RAS. The aim of this study was to compare the serum levels of total antioxidant status (TAS), nitric oxide (NO) and nitric oxide synthase (NOS) in minor RAS (MiRAS) patients with healthy individuals and determine the possible association of MiRAS with the 3 physiological parameters mentioned above. METHODS: Ninety patients with idiopathic MiRAS and 90 race-, age- and sex-matched healthy individuals were included in this study. All these subjects were allocated to 3 groups: MiRAS patients in the active stage (Group A); the same MiRAS patients in Group A in the inactive stage (Group B); healthy individuals without MiRAS (Group C). Serum levels of TAS, NO and NOS were determined by the spectrophotometric method. Independent sample t test and paired t test were performed for statistical evaluation. RESULTS: Serum TAS level of Group A was significantly decreased than that of Group C, whereas the serum level of NO was significantly higher in Group A as compared to Group C (P < .05). The serum levels of TAS and NO in Group B were no significant differences when compared with those in Group A or Group C. No significant differences in NOS activities were also found between the 3 groups (P > .05). CONCLUSIONS: MiRAS is associated with decreased TAS and increased NO levels, but NOS may not play an important role in the aetiopathogenesis.

Comparison and analysis of the therapeutic effect of different statins in the treatment of atherosclerosis.

Studies have confirmed that lipid-lowering drugs can effectively control the morbidity and mortality of cardiovascular and cerebrovascular diseases and statins are the most widely used. The aim of this study is to investigate the effect and mechanism of different statins on atherosclerotic patients. The patients were randomly divided into 4 groups according to the digital method, and the patients were treated with conventional therapy, simvastatin treatment, pravastatin treatment, atorvastatin treatment. It is concluded that statins are safe, effective and reliable for the treatment of atherosclerosis, and worthy of clinical promotion. The results also showed that after 6 months of taking statins, the levels of NO and NOS increased, the thickness of carotid intima-media became thinner and the plaque score decreased. This study provides a basis for elucidating the role of statins in the body.

Reduction in nitric oxide bioavailability shifts serum lipid content towards atherogenic lipoprotein in rats.

Nitric oxide (NO) is major endothelial relaxing factor and reduction in its bioavailabilty has been linked to hypertension. Furthermore, high lipid content is a strong risk factor predisposing to cardiovascular diseases. The principal focus of this study was to investigate the effect of blockade of nitric oxide synthase (NOS) on serum lipid content in rats. Male Wistar rats (150-170 g, n = 15) were randomly divided into two groups designated control (n = 5), and L-Name group (n = 10) and were gavage with distilled water and 60 mg/kg of L-NAME respectively daily for three weeks. After 3 weeks, the L-NAME group was sub-divided into two sub-groups (n = 5 each): L-NAME (60 mg/kg of L-NAME), and L-NAME plus ramipril (LR) (60 mg/kg of L-NAME plus 20 mg/kg of ramipril) and were treated daily for another three weeks. The blood pressure (BP) of the conscious rats was measured by tail-cuff method at the onset, at the third and at the sixth weeks of the experiment; while serum lipid contents and NO were measured at the third and sixth weeks. At the end of the experiment blood sample was drawn by ocular puncture for evaluation of lipid profile and NO, and the animals were later euthanized by overdose of anaesthesia. Data were analyzed using ANOVA at p < 0.05. There was a significant increase in BP, triglyceride, total cholesterol, low density lipoprotein-cholesterol, and atherogenic indices in L-NAME group compared to the control and LR group (p < 0.05); NO and high density lipoprotein-cholesterol was significant lower in the L-NAME group compared to control and LR (p < 0.05). In conclusion, reduction in NO bioavailability alters lipid metabolism, which was rectified by ramipril.

Ginkgetin ameliorates experimental atherosclerosis in rats.

Atherosclerosis is a common disease seriously detrimental to human health. Natural products are important sources of therapeutic candidates for atherosclerosis. We here evaluated the effects of ginkgetin on experimental atherosclerosis in rats and explored the underlying mechanisms. Atherosclerosis was induced by high-fat diet for 12 weeks combined with single intraperitoneal injection of vitamin D3 in rats. The atherosclerotic rats were then treated with ginkgetin at 25, 50 and 100 mg/kg/d or simvastatin at 2 mg/kg/d for 8 weeks. Blood and thoracic aortas were collected for analyses of histopathology, lipid deposition, serum biochemistry, matrix metalloproteinases (MMPs), and nitric oxide (NO)/NO synthase (NOS) system. We found that ginkgetin improved thoracic aortic intima structure, reduced intima-media thickness and intima/media ratio, and attenuated lipid deposition in aorta of atherosclerotic rats. Ginkgetin also decreased the serum levels of total cholesterol, triglyceride and low-density lipoprotein cholesterol, but restored the serum levels of high-density lipoprotein cholesterol in atherosclerotic rats. Additionally, ginkgetin reduced the mRNA and protein expression of MMP-2 and MMP-9 in thoracic aortas of rats with atherosclerosis. Further examinations showed that ginkgetin increased the NO and NOS levels in serum and thoracic aortas. Ginkgetin also unregulated the expression of endothelial NOS and downregulated the expression of inducible NOS at both mRNA and protein levels in thoracic aortas of atherosclerotic rats. Altogether, ginkgetin showed therapeutic effects on experimental atherosclerosis associated with improving lipid profile and modulating the MMPs and NO/NOS systems in rats. Ginkgetin could be a promising candidate for the treatment of atherosclerosis.

The effect of 131I-induced hypothyroidism on the levels of nitric oxide (NO), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), total nitric oxide synthase (NOS) activity, and expression of NOS isoforms in rats.

Accumulating evidence has shown that hypothyroidism affects the cardiovascular system, significantly increasing the incidence of cardiovascular diseases. In the present study we investigated the effect of radioactive iodine (I-131)-induced hypothyroidism on several parameters of vascular function, such as nitric oxide (NO), total nitric oxide synthase (NOS) activity and expression of NOS isoforms, as well as on interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) as indicators of inflammation, in rats. A dose of 150 µCi of 131-I was determined as optimal for establishing the model of hypothyroidism in rats. After administration of 131-I, at the end of month 1, 2, and 4 (n = 3 for each time point), NO, IL-6, and TNF-α in the serum and total NOS activity in the aorta were determined in 150 µCi group, compared to controls. The mRNA and protein expression of endothelial, neuronal, and inducible NOS (eNOS, nNOS, and iNOS) in the rat aorta was also estimated, using quantitative reverse transcription polymerase chain reaction and Western blot, respectively. The levels of IL-6 and TNF-α increased in 150 µCi group; the results were significant at the end of month 2 and 4 for IL-6, and at all time points for TNF-α. The levels of NO decreased significantly at the end of month 2 and 4 in 150 µCi group. The total NOS activity increased significantly in 150 µCi group, at all three time points. Significant changes in the mRNA and protein expression of all three NOS isoforms were observed in 150 µCi group compared to controls. NO, IL-6, TNF-α levels and NOS activity and expression are altered in hypothyroid state, and the underlying mechanism should be further investigated.

In Vivo Therapeutic Effect of Vaccinium Meridionale Swartz in Ischemia-Reperfusion Induced Male Albino Rats.

This study was aimed to investigate the cardioprotective and antioxidant effect of Vaccinium meridionale Swartz in ischemia-induced male albino Wistar strain rats. Rats were grouped into 5 of 6 numbers each. Group I served as a sham, group II served as control and group III, IV, and V served for 1, 10, and 25 mg/kg/d of an extract of Vaccinium meridionale Swartz for 15 consecutive days of treatment. Serum marker enzymes, lipid peroxidation, and myeloperoxidase were increased, whereas antioxidant enzymes were reduced in control due to injury. Increased phenol and anthocyanin contents and increased free radical scavenging activity was noted following treatment. Serum marker enzymes, necrosis, and lipid peroxidation, were reduced, whereas antioxidant enzymes and reduced glutathione were increased. Nitric oxide synthase and Akt expression were also increased following treatment. Taking all these data together, it is suggested that Vaccinium meridionale Swartz may be a potential therapeutic agent for the treatment of ischemic injury.

Spectrophotometric assays for measuring redox biomarkers in blood and tissues: the NADPH network.

Nicotinamide adenine dinucleotide (NAD+/NADH) along with its phosphorylated form (NADP+/NADPH) are two molecules ubiquitously present in all organisms, and they play key roles as cofactors in fundamental catabolic and anabolic processes, respectively. The oxidation of NADPH to NADP+ initiates a cascade of reactions, where a network of molecules is implicated. The molecules of this cascade form a network with eminent translational potential in redox metabolism. A special point of interest is that spectrophotometric assays have been developed both for NADH/NADPH and the molecules directly regulated by them. Therefore, crucial molecules of the NADPH-dependent redox network can be measured, and the results can be used to assess the bioenergetic and/or oxidative stress status. The main aim of this review is to collectively present the NADPH-related molecules, namely NADPH, NADH, NAD+ kinase, NADPH oxidase, peroxiredoxin, thioredoxin, thioredoxin reductase, and nitric oxide synthase, that can be measured in blood and tissues with the use of a spectrophotometer, which is probably the most simple, inexpensive and widely used tool in biochemistry. We are providing the researchers with reliable and valid spectrophotometric assays for the measurement of the most important biomarkers of the NADPH network in blood and other tissues, thus allowing the opportunity to follow the redox changes in response to a stimulus.

Anticoccidial effects of areca nut (Areca catechu L.) extract on broiler chicks experimentally infected with Eimeria tenella.

To study the anticoccidial effects of areca nut extract (ANE) on coccidiosis, 270 one-day old Wenchang broiler chicks were divided into six equal groups, each with three replicate cages (n = 15 per cage). The six groups were the blank control group (BC), negative control group (NC), positive control group (PC), and three ANE-treated groups. The birds in the three control groups (BC, NC and PC) were fed a basal diet without ANE supplementation. The birds in the three ANE-treated groups were fed a basal diet supplied with ANE at 100 (T1), 200 (T2), or 300 (T3) mg/kg feed. At 15 days of age, the birds in the NC, PC and the three ANE groups were challenged orally with 1 × 105Eimeria tenella oocysts per chick. At 48 h after oocysts inoculation, the birds in group PC were supplied diclazuril with drinking water for 5 days. The results showed that ANE and diclazuril significantly improved feed intake and body weight gain (P < 0.05) relative to the NC group. Both ANE and diclazuril significantly (P < 0.05) reduced OPG on day 4-9 post-inoculation (p.i.) relative to the NC group. Coccidial infection damaged the integrity of the cecal mucosa and thickened cecal tunica muscularis. ANE and diclazuril mitigated the mucosal damage caused by coccidial infection. Diet ANE supplementation reduced the cecal lesion scores compared to the NC group (P < 0.05). ANE and diclazuril increased nitric oxide (NO) levels at 3 days p.i., but reduced NO levels at 6 days p.i. (P < 0.05) compared to the NC group. Diet ANE supplementation increased the concentration of interleukin 2 (IL-2) in infected chicken relative to the NC group. The current results showed the anticoccidial properties, and beneficial effect on intestinal mucosa damage of ANE in broiler chicks challenged with coccidiosis.

Oxidative and Nitrosative Stress as Well as the Tryptophan Catabolites Pathway in Depressive Disorders.

The aim of this paper is to elucidate the role of oxidative and nitrosative stress as well as the tryptophan catabolites pathway in the development of depression and the mechanism of action of antidepressant drugs, based on the available literature. According to the World Health Organization (WHO), an estimated 350 million people worldwide suffer from depression. The pathogenesis of depressive disorders has not been fully explained yet and several causes of this disease have been suggested. There is evidence for the involvement of several interconnected biochemical pathways, including oxidative and nitrosative stress as well as the tryptophan catabolites pathway. Studies to date indicate that patients with depression have lower levels of enzymatic and non-enzymatic elements of an antioxidant response and, at the same time, they display an increased amount of oxidative stress markers, when compared to healthy individuals. The development of depression is also associated with excessive activity of nitric oxide synthase. Furthermore, decreased levels of tryptophan and increased levels of its harmful catabolites, i.e. kynurenine and quinolinic acid, may lead to progression of the disease. Changes in these biochemical pathways can be used as risk factors for the development of depression and, in the future, they could be utilized as diagnostic biomarkers. Moreover, regulation of biochemical processes may contribute to the development of a new, effective and personalized antidepressant therapy.

[Effects of Different Acupuncture and Moxibustion Methods on Plasma NO, NOS and VIP Contents and the Expression of Colonic VIP Protein in Functional Constipation Rats].

OBJECTIVE: To compare the effects of filiform needling, electroacupuncture, and moxibustion on functional constipation in rats, and to explore the possible mechanism of these three different methods. METHODS: Male SD rats were randomly divided into blank control group (n=8), model group (n=11), medication group (n=8), filiform needling group (n=11), electroacupuncture group (n=11), and moxibustion group (n=11). Functional constipation model was established by intragastric administration with the suspension of loperamide hydrochloride, daily for six days in a week. One hour after each gavage, the medication group was treated with cisapride suspension, while the other three groups were treated with filiform needling, electroacupuncture, and moxibustion, respectively, at "Tianshu"(ST 25), "Shangjuxu" (ST 37) acupoints. The first defecation time was recorded. NO, NOS and VIP (vasoactive intestinal peptide) levels in plasma were detected by ELISA. Immuohistochemical and Western blot methods were applied to test VIP expression in the colonic tissue. RESULTS: Compared with the blank control group, the first defecation time was prolonged, and the NO, NOS and VIP contents in plasma were increased in the model group (P<0.05, P<0.01), accompanied with increased score of VIP in the smooth muscle of colonic wall(P<0.05)and elevated VIP expression in colonic tissue(P<0.01). Compared with the model group, the first defecation time was shortened in the medication, filiform needling, electroacupuncture, and moxibustion groups(P<0.05), while decreased NO, NOS and VIP contents in plasma (P<0.01,P<0.05), decreased score of VIP in the smooth muscle of colonic wall(P<0.05), and lower expression level of VIP in colonic tissue(P<0.01)were observed in the four treatment groups. In contrast, the plasma VIP content in the electroacupuncture group was lower than that in the medication group (P<0.01), while colonic VIP expression decreased in the moxibustion group(P<0.01). The NO, NOS and VIP contents in plasma and score of VIP in the smooth muscle of colonic wall in the electroacupuncture group were lower than those in the filiform needling group and the moxibustion group(P<0.01, P<0.05), but the colonic VIP expression in the moxibustion group was lower than that in the filiform needling group(P<0.05)and electroacupuncture group(P<0.01). CONCLUSIONS: All the three different kinds of acupuncture and moxibustion methods have positively regulatory effect on functional constipation; electroacupuncture is the best for the regulation of plasma NO, NOS and VIP contents, while moxibustion is the best for the regulation of VIP expression in colonic tissue.

Decreased serum L-arginine and L-citrulline levels in major depression.

RATIONALE: It has been suggested that endothelial dysfunction caused by a decreased endothelial production of nitric oxide (NO) may contribute to the consistently observed increased risk of developing cardiovascular disease (CVD) in physically healthy patients suffering from major depression (MD). NO is a gas synthesized from Larginine (a conditionally essential amino acid) and oxygen by endothelial nitric oxide synthase (eNOS). The end products of NO production include both NO and L-citrulline. NO is rapidly reduced to the anions nitrite and nitrate, classically referred to as NO metabolites. Their measurement has been used as a surrogate measurement for endothelial NO production. We and others have shown decreased levels of NO metabolites in the serum of MD patients. The mechanism of this decreased production of NO by the endothelium has not yet been elucidated. OBJECTIVES: The purpose of this study is to assess serum levels of L-arginine and L-citrulline in patients with MD. METHODS: Levels of L-arginine and L-citrulline were measured in 35 unmedicated physically healthy MD patients and 36 healthy controls (HCs). RESULTS: L-arginine and L-citrulline concentrations were significantly lower in MD patients than in healthy controls (L-arginine, 73.54 + 21.53 µmol/L and 84.89 + 25.16, p = 0.04 µmol/L and L-citrulline 31.58 + 6.05 µmol/L and 35.19 + 6.85 µmol/L, p = 0.03, respectively). CONCLUSIONS: The decrease in L-arginine levels in MD patients is a possible explanation for the decrease in NO metabolites in MD patients and therefore may contribute, through endothelial dysfunction, to the increased CV risk associated with MD.

Plasmid pLXSN-Mediated Adrenomedullin Gene Therapy for Cerebral Vasospasm Following Subarachnoid Hemorrhage in Rats.

BACKGROUND The aim of this study was to investigate the protective effect of ADM gene mediated by plasmid pVAX1 on cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH). MATERIAL AND METHODS The recombinant plasmid pVAX-ADM was successfully established, and 40 SD rats were randomly divided into normal saline, pVAX1, pVAX1-ADM low-dose, pVAX1-ADM mid-dose, and pVAX1-ADM high-dose groups. The circumference and diameter of basilar artery, diameter of middle cerebral artery and internal carotid artery, and thickness of basilar artery wall were observed. The levels of circulating endothelial cells (CEC) and levels of regional cerebral blood flow (rCBF) of the parietal cortex were detected at different time-points. The expression levels of serum ADM, ET-1, and NOS of each group and the neurological functions were compared. RESULTS The circumference and diameter of basilar artery and the diameter of the middle cerebral artery and internal carotid artery in pVAX1-ADM groups were significantly longer than those in the saline group and pVAX1 group (P<0.05), but the thickness of the basilar artery wall in pVAX1-ADM groups was significantly lower (P<0.05), and the levels of growth or decrease were both dose-dependent (P<0.05). Compared with the saline group and pVAX1 group, the expression levels of serum ADM, NOS, and rCBF in pVAX1-ADM groups were significantly higher (P<0.05), but the levels of serum ET-1 and CEC were significantly lower (P<0.05). The scores of neurobehavioral functions of pVAX1-ADM groups were significantly lower (P<0.05), and the scores were also dose-dependent (P<0.05). CONCLUSIONS The recombinant eukaryotic expression plasmid pVAX1-ADM can significantly relieve cerebral vasospasm, increase the expression of serum ADM and NOS, and decrease the expression of serum ET-1 in a rat model of CVS; it is dose-dependent and can also improve nervous system function.

Asymmetric dimethylarginine and all-cause mortality: a systematic review and meta-analysis.

Asymmetrical dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase (NOS), impairs the beneficial effect of NO. The predictive value of ADMA for all-cause mortality remains controversial, though it is important in the development of cardiovascular disease (CVD) and progression to dialysis in renal disease. This systematic review and meta-analysis was conducted to investigate the association between circulating ADMA and all-cause mortality. Studies with data pertinent to the association between circulating ADMA and all-cause mortality were reviewed and OR, HR or RR with 95% CI derived from multivariate Cox's proportional-hazards analysis were extracted. A total of 34 studies reporting 39137 participants were included in final analysis. The results demonstrated that circulating ADMA was independently associated with all-cause mortality (RR = 1.27, 95% CI: 1.20-1.34). The association was still statistically significant in patients with pre-existing renal disease (RR = 1.30, 95% CI: 1.19-1.43) and pre-existing CVD (RR = 1.26, 95% CI: 1.16-1.37). In those without pre-existing renal or CVD, ADMA also predicted all-cause mortality (RR = 1.31, 95% CI: 1.13-1.53). The present study suggests a positive association of circulating ADMA with all-cause mortality. Further studies are needed to investigate the effects of interventions on ADMA, and the value of ADMA as a biomarker.

Total Glucosides of Paeony Promote Intestinal Motility in Slow Transit Constipation Rats through Amelioration of Interstitial Cells of Cajal.

OBJECTIVES: Using an atropine-diphenoxylate-induced slow transit constipation (STC) model, this study explored the effects of the total glucosides of paeony (TGP) in the treatment of STC and the possible mechanisms. STUDY DESIGN: A prospective experimental animal study. METHODS: The constipation model was set up in rats with an oral gavage of atropine-diphenoxylate and then treated with the TGP. The volume and moisture content of the faeces were observed and the intestinal kinetic power was evaluated. Meanwhile, the colorimetric method and enzyme linked immunosorbent assay (ELISA) were employed to determine the changes of nitric oxide (NO), nitric oxide synthase (NOS), vasoative intestinal peptide (VIP) and the P substance (SP) in the serum, respectively. The protein expressions of c-kit and stem cell factor (SCF) were assessed by immunohistochemical analysis and western blot, respectively, and the mRNA level of c-kit was measured by a reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The TGP attenuated STC responses in terms of an increase in the fecal volume and moisture content, an enhancement of intestinal transit rate and the reduction of NO, NOS and VIP in the serum. In addition, the c-kit, a labeling of interstitial cells of Cajal (ICC) increased at both protein and mRNA levels. SCF, which serves as a ligand of c-kit also increased at protein level. CONCLUSION: The analysis of our data indicated that the TGP could obviously attenuate STC through improving the function of ICC and blocking the inhibitory neurotransmitters such as NO, NOS and VIP.

Reversibility of endothelial dysfunction in diabetes: role of polyphenols.

The endothelium, a thin single sheet of endothelial cells, is a metabolically active layer that coats the inner surface of blood vessels and acts as an interface between the circulating blood and the vessel wall. The endothelium through the secretion of vasodilators and vasoconstrictors serves as a critical mediator of vascular homeostasis. During the development of the vascular system, it regulates cellular adhesion and vessel wall inflammation in addition to maintaining vasculogenesis and angiogenesis. A shift in the functions of the endothelium towards vasoconstriction, proinflammatory and prothrombic states characterise improper functioning of these cells, leading to endothelial dysfunction (ED), implicated in the pathogenesis of many diseases including diabetes. Major mechanisms of ED include the down-regulation of endothelial nitric oxide synthase levels, differential expression of vascular endothelial growth factor, endoplasmic reticulum stress, inflammatory pathways and oxidative stress. ED tends to be the initial event in macrovascular complications such as coronary artery disease, peripheral arterial disease, stroke and microvascular complications such as nephropathy, neuropathy and retinopathy. Numerous strategies have been developed to protect endothelial cells against various stimuli, of which the role of polyphenolic compounds in modulating the differentially regulated pathways and thus maintaining vascular homeostasis has been proven to be beneficial. This review addresses the factors stimulating ED in diabetes and the molecular mechanisms of natural polyphenol antioxidants in maintaining vascular homeostasis.

Platelet hyperaggregability in obesity: is there a role for nitric oxide impairment and oxidative stress?

Epidemiological evidence has shown that platelet activation markers are consistently elevated in obesity, contributing to its prothrombotic state. In order to improve the understanding of the regulation of platelet function in obesity, the aim of this study was to investigate the l-arginine-nitric oxide (NO) pathway in obese adults without other cardiovascular risk factor. Seventeen obese (body mass index [BMI] 35.9±1.0 kg/m(2) ) and eighteen age-matched normal weight subjects (BMI 22.0±0.6 kg/m(2) ) were included in this study. l-arginine influx was measured with incubation of l-[(3) H]-arginine. NO synthase (NOS) and arginase activities were determined by the citrulline assay and the conversion of l-[(14) C]-arginine to [(14) C]-urea, respectively. Cyclic guanosine monophosphate (cGMP) content was evaluated by enzyme-linked immunosorbent assay. In addition, the study analyzed: platelet aggregation; intraplatelet antioxidant enzymes, via superoxide dismutase (SOD) and catalase activities; and systemic levels of l-arginine, fibrinogen, and C-reactive protein (CRP). Obese patients presented a significant decrease of platelet l-arginine influx, NOS activity, and cGMP levels, along with platelet hyperaggregability. On the presence of NO donor, platelet aggregation was similar between the groups. The fibrinogen and CRP systemic levels were significantly higher and SOD activity was reduced in obesity. No significant differences were observed in plasma levels of l-arginine and intraplatelet arginase and catalase activities between groups. The diminished NO bioavailability associated with inflammatory status and impaired enzymatic antioxidant defence may contribute to future cardiovascular complications in obesity.