OBJECTIVETo explore the possible mechanism of the erector spinal muscles in idiopathic scoliosis by comparing the expression and localization of neuronal nitric oxide synthase (nNOS) and inducible nitric oxide synthase (iNOS) of the thoracic erector spinal muscles on convex side and concave side.

METHODSThe patient group comprised 8 females and 2 males who were scheduled for spinal surgery. The apex of scoliotic curve in these patients arose between T6 and T11. The mean age was 14.3 (range 12-17) years, and the mean Cobb angle was 57.7 degrees (range 45 degrees-85 degrees). Muscle biopsies were taken bilaterally during surgery from the superficial multifidus muscle at the apex of the curve between the 6th and 11th thoracic vertebral levels. Part of the tissue was fixed in formalin and stained with hematoxylin and eosin; the remaining tissue was snap frozen and processed for immunohistochemistry and Western blot. Immunocytochemistry for nNOS and iNOS were performed using the EnVision two-step method. Western blot was done with antibodys to nNOS and iNOS. Immunoreactive bands were visualized by enhanced chemiluminescence according to the manufacturer's specifications (Amersham Corp).

RESULTSnNOS protein in the erector spinal muscles was localized at the sarcolemma. Western blot demonstrated that nNOS protein expression in the concave side of erector spinal muscles is more than that in the convex side. A significant decrease in nNOS protein and activity was found on the convex side of erector spinal muscles from idiopathic scoliosis patients; There was a little immunoreactivity to iNOS in erector spinal muscles. There was little difference in iNOS protein expression between both sides of the curve. Western blot detected the same results.

CONCLUSIONThere is a greater expression of nNOS and iNOS on the concave side than on the convex side, suggesting nNOS and iNOS may play a role in the pathogenesis of idiopathic scoliosis.

Adolescent ; Child ; Female ; Humans ; Immunohistochemistry ; Male ; Muscle, Skeletal ; cytology ; enzymology ; Nitric Oxide Synthase ; analysis ; metabolism ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type II ; Scoliosis ; enzymology

BACKGROUNDThe form deprivation (FD) reduces spatial contrasts and induces myopia. Nitric oxide and cyclic guanosine monophosphate (cGMP) are involved in visual signal transmission. This study investigated changes in nitric oxide synthase (NOS) activity and cGMP concentration in ocular tissues in acute and chronic form deprivation myopia.

METHODSGuinea pigs had one eye covered by translucent glass for 7, 14 or 21 days. Untreated litter mates were used as controls. NOS activity and cGMP concentrations in the retinal, choroidal and scleral tissues of FD eyes and control eyes were analyzed by radioimmunoassay after various durations of FD. The expression of NOS subtypes was identified by immunohistochemistry.

RESULTSMyopia was successfully induced in FD eyes after 14 days. Compared with control groups, the retinal NOS activity and cGMP concentrations in the FD eyes significantly increased after 14 and 21 days while the retinal NOS activity in the FD eyes was transiently suppressed by 7 days of FD. The NOS activity and cGMP concentrations of choroid and sclera in the FD eyes were higher than in the control groups at 21 days. The three isoenzymes of nitric oxide synthase were detected in the ocular tissues of guinea pigs.

CONCLUSIONSThe NOS activity and cGMP concentrations were upregulated after chronic FD and the retinal NOS activity was transiently suppressed at acute FD. The function of elevated NOS activity may be mediated by cGMP.

Animals ; Cyclic GMP ; analysis ; Guinea Pigs ; Immunohistochemistry ; Myopia ; metabolism ; Nitric Oxide ; physiology ; Nitric Oxide Synthase ; metabolism ; Refractive Errors ; Retina ; metabolism

BACKGROUNDTo date, there have been no reports on altered nitric oxide (NO) content in ischemia/reperfusion with regard to in vivo preconditioning procedures. These studies are important for understanding the mechanisms of NO during early myocardial ischemic preconditioning. The aim of the present study was to investigate the mechanisms of NO during early myocardial ischemic preconditioning by measuring levels of NO and cyclic guanosine monophosphate (cGMP), as well as activity of nitric oxide synthase (NOS) in ischemia/reperfusion with respect to preconditioning in rats.

METHODSSixty-six female Sprague-Dawley rats were randomly divided into four groups: ischemic preconditioning group (IP), ischemia/reperfusion group (I/R), control group (CON), and preconditioning procedure group (PC). In the PC group, rats were further divided into PC1-, PC1 +, PC2-, PC2 +, PC3-, and PC3 + subgroups. Rats underwent left coronary artery occlusion and reperfusion, and subsequently, NOS activity and levels were assessed with spectrophotometric analysis. cGMP contents were measured with radioimmunoassay.

RESULTSThe level of NO and cGMP, as well as the activity of NOS, were significantly higher in the IP group compared to the I/R and CON groups (P < 0.05). During preconditioning prior to prolonged ischemia, NO and cGMP levels varied markedly with ischemia and reperfusion. The levels of NO repeatedly increased when the heart was exposed to three episodes of 5-minute ischemia, and were almost completely reversed during each reperfusion period. NO and cGMP levels were significantly different between the 5-minute period of ischemia and the same period of reperfusion during preconditioning.

CONCLUSIONSNO plays an important role during early phase myocardial ischemic preconditioning in rats. NO and cGMP could be triggers and mediators of early phase myocardial ischemic preconditioning. Altered NOS activity following ischemic stress could be the primary inducer of higher NO levels detected. NO and cGMP fluctuations might be the trigger for protection during early phase myocardial ischemic preconditioning.

Animals ; Cyclic GMP ; analysis ; Female ; Ischemic Preconditioning, Myocardial ; Nitric Oxide ; analysis ; physiology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship of nitric oxide (NO) and nitric oxide synthase (NOS) with varicocele (VC) infertility.

METHODSFifty-three infertile men, 21 with varicocele and 32 with subvaricocele, were enrolled as Group 1, 29 infertile patients with oligoasthenozoospermia but without varicocele as Group 2 and 28 normal fertile controls as Group 3. The NO content and NOS activity in the seminal plasma and peripheral blood were measured by nitric acid reductase method, and the semen parameters of VC determined by computer-assisted semen analysis (CASA).

RESULTSSignificant differences were noted between Group 1 and the other two in the NO content and NOS activity in the seminal plasma (P < 0.05) but not in the peripheral blood (P > 0.05). In Group 1, the NO content and NOS activity were increased in both the seminal plasma and peripheral blood with the enhanced diameter of the varicose spermatic vein, with a significant difference only in the seminal plasma between the varicocele and subvaricocele patients (P < 0.05), and the same increase was observed with decreased sperm concentration (> or = 20 x 10(6)/ml and < or = 10 x 10(6)/ml) and motility (> or = 50% and < or = 25%), with significant differences (P < 0.05).

CONCLUSIONNO plays an important role in the VC-induced decrease of seminal quality. For the diagnosis of VC, the determination of the NO content and NOS activity in the seminal plasma is of more significance than that in the peripheral blood, and the earlier the determination, the greater its clinical value for both the diagnosis and treatment of VC.

Adult ; Humans ; Infertility, Male ; etiology ; metabolism ; Male ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Semen ; cytology ; metabolism ; Semen Analysis ; Sperm Count ; Varicocele ; complications ; metabolism ; Young Adult

OBJECTIVETo explore the mischief mechanism of oxidative stress in the varicocele (VC).

METHODSSerum was taken from the spermatic and peripheral veins on ligation of the internal spermatic veins in 28 infertile males with VC. Experimental VC was established in male rats by partial ligation of the left renal vein. And testis tissue was taken three months after operation. The nitric oxide(NO), nitric oxide synthetase (NOS), xanthine oxidase (XO), lactic acid(Lac) and lactic dehydrogenase (LDH) in the serum of 28 infertile males with VC and the testis tissue of the VC rats were detected by spectrophotometry.

RESULTSNO, NOS, XO and Lac in the serum of internal spermatic veins in the infertile males with VC were significantly higher than in the serum of peripheral veins in the VC patients (P < 0.01, P < 0.05). LDH was lower than that in peripheral serum. NO and XO of the left testis tissue in the VC rats were higher compared with the control group (P < 0.01). Lac in the left testis of the VC rats was lower than that in the control group rats (P < 0.01).

CONCLUSIONNO, NOS and XO in the serum of the VC patients and in the testis tissues of the VC rats were increased, and Lac and LDH were changed obviously, which might not only disturb spermatogenesis, but also inhibit sperm motility. Therefore they might be one of the causes of infertility in VC patients.

Adult ; Animals ; Humans ; Infertility, Male ; etiology ; Male ; Nitric Oxide ; analysis ; Nitric Oxide Synthase ; analysis ; Oxidative Stress ; Rats ; Rats, Wistar ; Varicocele ; complications ; metabolism ; Xanthine Oxidase ; analysis

OBJECTIVETo probe into the effect of acupuncture at "Sishencong" (EX-HN 1) on sleep and the mechanism.

METHODSForty-eight white mice were randomly divided into a Sishencong group, a Zusanli group, a model group and a blank control group, 12 mice in each group. In the mice of the first 3 groups, sleep rhythm disorders was induced by repeatedly regular intraperitoneal injection of small dose of Caffeine, and the mice of the first two groups were treated by acupuncture at "Sishencong" (EX-HN 1) and "Zusanli" (ST 36), respectively. Living state and, circadian activities were observed, and nitric oxide synthase (NOS) activity and nitric oxide(NO) content in the brain were detected.

RESULTSAcupuncture at "Sishencong" (EX-HN 1) or "Zusanli" (ST 36) could significantly increase NOS activity and NO content in the brain, with acupuncture at "Sishencong" (EX-HN 1) being significantly better than acupuncture at "Zusanli" (ST 36) (P < 0.01).

CONCLUSIONAcupuncture at Sishencong (EX-HN 1) treats insomnia and regulates sleep and many mental and nervous symptoms, which are carried out partially through increasing NOS activity and NO content, promoting normal function of brain tissues.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Brain ; metabolism ; Disease Models, Animal ; Female ; Male ; Mice ; Nitric Oxide ; analysis ; Nitric Oxide Synthase ; metabolism ; Sleep Wake Disorders ; physiopathology ; psychology ; therapy

BACKGROUNDThis study was designed to investigate the potential pathogenicity of Mycoplasma penetrans (M. penetrans) and its molecular mechanisms responsible for the induction of iNOS gene expression in mouse macrophages stimulated by lipid-associated membrane proteins (LAMPs) prepared from M. penetrans.

METHODSMouse macrophages were stimulated with M. penetrans LAMPs to assay the production of nitric oxide (NO). The expression of inducible nitric oxide synthase (iNOS) was detected by RT-PCR and Western blotting. The activity of nuclear factor kappaB (NF-kappaB) and the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappaB, on the production of nitric oxide and the expression of iNOS were also assessed in mouse macrophages treated with M. penetrans LAMPs by indirect immunofluorescence and Western blotting.

RESULTSM. penetrans LAMPs stimulated mouse macrophages to produce nitric oxide in a dose- and time-dependent manner. The mRNA and protein levels of iNOS were also upregulated in response to LAMP stimulation and inhibited by PDTC treatment. M. penetrans LAMPs were found to trigger NF-kappaB activation, a possible mechanism for the induction of iNOS expression.

CONCLUSIONThis study demonstrated that M. penetrans may be an important etiological factor of certain diseases due to the ability of M. penetrans LAMPs to stimulate the expression of iNOS, which is probably mediated through the activation of NF-kappaB.

Animals ; Bacterial Proteins ; pharmacology ; Cells, Cultured ; Enzyme Induction ; Lipoproteins ; pharmacology ; Membrane Proteins ; pharmacology ; Mice ; Mycoplasma penetrans ; chemistry ; NF-kappa B ; metabolism ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; biosynthesis ; Nitric Oxide Synthase Type II ; RNA, Messenger ; analysis

OBJECTIVEImmunohistochemistry using antibodies to dystrophin is the pathological basis for the differential diagnosis of Duchenne and Becker muscular dystrophy (DMD and BMD). In rare cases, however, labelling dystrophin on sarcolemma is equivocal and similar to that observed in controls. This makes the diagnosis of BMD difficult. This study aimed to explore the diagnostic value of neuronal nitric oxide synthase (nNOS) antibody for clinically suspected BMD.

METHODSImmunohistochemical staining was performed on muscle specimens of 5 cases of BMD with positive expression of Dys-C (3 cases had a confirmed diagnosis of BMD, 2 cases were clinically suspected as BMD) by using dystrophin and nNOS antibodies. Normal muscle specimens from the children with fracture were used as controls.

RESULTSCompared with the controls, the expression of Dys-R, Dys-C and Dys-N was markedly reduced and nNOS was not expressed on sarcolemma in the three cases of definitely diagnosed BMD. The two cases of clinically suspected as BMD had a complete absence of sarcolemmal nNOS, even if had a similar expression of dystrophin on sarcolemma to the controls.

CONCLUSIONSnNOS antibody staining can be used for a definite diagnosis in children with clinically suspected BMD who have the almost normal expression of dystrophin.

Child ; Child, Preschool ; Dystrophin ; analysis ; chemistry ; Humans ; Immunohistochemistry ; Infant ; Muscular Dystrophy, Duchenne ; diagnosis ; metabolism ; Nitric Oxide Synthase Type I ; analysis

Angiotensins ; analysis ; Animals ; Animals, Newborn ; Biomarkers ; analysis ; Endothelin-1 ; analysis ; Hypertension, Pulmonary ; drug therapy ; metabolism ; physiopathology ; Lung ; chemistry ; pathology ; Magnesium Sulfate ; pharmacology ; Nitric Oxide Synthase ; analysis ; Nitric Oxide Synthase Type II ; Swine ; Vasomotor System ; chemistry

OBJECTIVETo study the levels of nitric oxide synthase (NOS) and nitric oxide (NO) in patients with coronary heart disease (CHD), and the factors affecting them.

METHODSConcentrations of NO and the activities of NOS in plasma in 136 patients and 206 controls using the corresponding Kits were measured. The data were analyzed using covariance and multiple linear regression analysis with SAS 8.1.

RESULTSThe levels of NO [(217.05 +/- 153.31) micromol/L] and NOS [(14.09 +/- 7.14) U/ml] in patients were significantly higher than that in the healthy controls [(140.69 +/- 90.96) micromol/L, (7.75 +/- 3.79) U/ml, respectively, P < 0.01]. Smoking and drinking were the independent risk factors for NOS, while sex was the independent risk factor for NO (P < 0.01).

CONCLUSIONSThe levels of plasma NO and NOS are closely related with coronary heart disease.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Coronary Disease ; blood ; Female ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Nitric Oxide ; blood ; Nitric Oxide Synthase ; blood ; Plasma ; metabolism