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

OBJECTIVESTo determine the expression of nitric oxide synthase (NOS) in testis and to investigate the effects of NO on the reproductive function of testis.

METHODSTestes of adult male Sprague-Dawley rats were fixed in 4% paraformaldehyde. The paraffin sections were made as routine. Immunohistochemical ABC method was used to observe the localization of NOS.

RESULTSEndothelia NOS (eNOS), neuronal NOS (nNOS) and inductive NOS (iNOS) were all expressed in Leydig cells. Only eNOS was expressed in peritubular myoid cells, endothelial and smooth muscle cells of blood vessel, while only nNOS expressed in tunica adventitia of testicular blood vessels. The reactive substance distributes in cytoplasm with negative nuclei. Immunoreactivity for eNOS, nNOS and iNOS in all spermatogenic cells was negative.

CONCLUSIONSThree kinds of NOS were all expressed in testis and the distribution of different NOS had a little difference.

Animals ; Immunohistochemistry ; Male ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; analysis ; Rats ; Rats, Sprague-Dawley ; Testis ; enzymology

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

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

The expression of both constitutive and inducible forms of nitric oxide synthase (NOS) was investigated by immunohistochemical staining of formalin-fixed paraffin-embedded sections in normal and Listeria monocytogenes-infected brains of goats. In normal control goats, a small number of neurons showed immunoreactivity of both iNOS and nNOS, and the number of iNOS-positive neurons was higher than the number of nNOS-positive neurons. In natural listeriosis, listeria antigens were easily immunostained in the inflammatory cells of microabscesses. In this lesion, the immunoreactivity of iNOS in neurons was more intense than the control, but nNOS was not. In microabscesses, nNOS was weakly visualized in macrophages and neutrophils, while iNOS was expressed in macrophages, but not in neutrophils. These findings suggest that normal caprine brain cells, including neurons, constitutively express iNOS and nNOS, and the expressions of these molecules is increased in Listeria monocytogenes infections. Furthermore, inflammatory cells, including macrophages, expressing both nNOS and iNOS may play important roles in the pathogenesis of bacterial meningoencephalitis in goat.
Animals ; Brain/cytology/*enzymology ; Glial Fibrillary Acidic Protein/analysis ; Goat Diseases/*enzymology ; Goats ; Immunohistochemistry ; Listeria Infections/enzymology/*veterinary ; Neurons/*enzymology ; Nitric Oxide Synthase/*analysis ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type II

OBJECTIVETo investigate protein and mRNA expression of nitric oxide synthase (NOS) isoforms I and III in rat penile tissues.

METHODSPenile tissues from male SD rats were investigated. The protein expression of NOS I and III in rat penis was confirmed by immunoprecipitation and Western blot. Gene expression of both NOS I and III was identified by reverse transcription-polymerase chain reaction.

RESULTSThe mRNA and protein products of NOS I and III are both expressed in normal rat penile spongium tissues. But, NOS III mRNA and protein products are more highly expressed than those of NOS.

CONCLUSIONSThese findings support the possibility that penile erection is regulated by different NOS isoforms in rat penile tissues.

Animals ; Blotting, Western ; Male ; Nitric Oxide Synthase ; analysis ; genetics ; Nitric Oxide Synthase Type I ; Nitric Oxide Synthase Type III ; Penis ; enzymology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

PURPOSE: This study was performed to investigate the expression pattern of inducible nitric oxide synthase (iNOS) in osteosarcoma and to determine whether its expression correlates with metastatic potential and prognosis. MATERIALS AND METHODS: Pathologic tissues obtained from 48 patients with primary osteosarcoma were examined immunohistochemically.Stained tissues were categorized into three groups (negative, mild, strong), according to expression intensity, and matched with clinical data. Results were analysed statistically. RESULTS: iNOS was expressed in 18 of 48 samples (38%). Among the iNOS positive cases, mild expression was observed in 9 patients (18%). There was statistically significant correlation between mild iNOS expression and metastasis (p=0.019). By multivariate analysis mild iNOS expression turned out to be a significant prognostic factor for reduced survival (Risk ratio 4.399, 95% CI 1.694-11.421: p=0.02). CONCLUSION: Mild expression of iNOS was significantly related to metastasis in osteosarcoma and can be considered a significant prognostic factor.
Humans ; Immunohistochemistry ; Multivariate Analysis ; Neoplasm Metastasis ; Nitric Oxide Synthase Type II* ; Osteosarcoma* ; Prognosis* ; Survival Analysis

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

BACKGROUNDIn China the ginseng root began to be used in medicine over 2000 years ago. Ginsenosides are the most important component isolated from ginseng. The authors investigated the effect of ginsenoside Rg1 on the spectrum of gene expression in the endothelial cells stimulated by TNF-alpha and further explored the potential molecular mechanism of endothelial protection by ginsenoside Rg1.

METHODSNitric oxide (NO) production in the cultured human umbilical vein endothelial cells (HUVECs) was measured by using an NO assay kit. A home-made oligonucleotide microarray containing approximately 400 cardiovascular disease-related genes was constructed. The alteration of the spectrum of gene expression induced by ginsenoside Rg1 in HUVECs which were activated by TNF-alpha were detected by oligonucleotide microarray analysis.

RESULTSNO production in HUVECs was decreased significantly after TNF-alpha treatment, while pretreatment with ginsenoside Rg1 enhanced NO production in TNF-alphastimulated HUVECs. Ginsenoside Rg1 affected the expression levels of genes involved in vascular constriction, cell adherence, coagulation, cell growth and signal transduction in TNF-alphastimulated HUVECs.

CONCLUSIONSGinsenoside Rg1 could enhance NO production and the expression of eNOS mRNA in TNF-alpha stimulated HUVECs. Ginsenoside Rg1 regulated sets of genes in endothelial cells and protected endothelial cells from TNF-alpha activation. Microarray analysis provided us with valuable insights into the atheroprotective mechanism by gingsenoside Rg1.

Endothelial Cells ; drug effects ; physiology ; Gene Expression ; drug effects ; Ginsenosides ; pharmacology ; Humans ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; analysis ; Nitric Oxide Synthase Type III ; Oligonucleotide Array Sequence Analysis ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the effects of super painkiller on the sexual function of male Wistar rats with diabetes.

METHODSThirty-two Wistar rats at 3-5 months of age were divided into two groups at random: 22 in the diabetes group, and 10 in the control group. After 72 hours, the former was further divided into 2 subgroups: non-treatment and super painkiller treatment. In 5 weeks, blood glucose was determined. After the apomorphines experiment, the rats were killed, blood taken from the vein, homogenate prepared from the isolated testis tissue and the level of NO and NOS in the serum and tissue homogenate surveyed, separately.

RESULTSCompared with the control group, serum NO and NOS of the diabetic rats dropped sharply. Compared with the non-treatment group, the serum NO and NOS of the super painkiller treatment group were very high and the difference was significant (P < 0.01). Apomorphine injection showed that the times of penis erection of the treated rats were more than those of the non-treatment group (P < 0.01) , but the difference was not significant compared with the control (P > 0.05).

CONCLUSIONSuper painkiller has sure effect of reducing blood glucose, with a certain curative value for sexual and reproductive malfunction of diabetic rats.

Animals ; Diabetes Mellitus, Type 1 ; complications ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Nitric Oxide ; analysis ; Nitric Oxide Synthase ; analysis ; Rats ; Rats, Wistar ; Sexual Dysfunctions, Psychological ; drug therapy