Objective To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion. Methods Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics. Results The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining. Conclusions The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.
ObjectiveTo explore the predictive value of fractional exhaled nitric oxide (FeNO) in the treatment response of adult asthmatic patients. Methods64 adult outpatients with asthma from Peking Union Hospital between March and September 2013 were recruited in the study. All patients completed asthma control test (ACT) together with exhaled nitric oxide (FeNO) and pulmonary function test. Then the patients were classified into a higher FeNO group (n=33) and a normal FeNO group (n=31) according to FeNO level. All patients accepted regular inhaled ICS/LABA treatment (salmeterol and fluticasone 50/250). Three months later all patients reaccepted ACT,FeNO and pulmonary function test. ResultsThe ACT score increased in all patients,and was significantly higher in the higher FeNO group than that in the normal FeNO group[22.07±5.49 vs. 19.23±5.48,t=2.893,P<0.05]. The complete control rate of the higher FeNO group was higher than that in the normal FeNO group (42.42% vs. 19.35%,χ2=3.960,P<0.05). The FEV1 and FEV1%pred of two groups both increased significantly (P<0.05),but there was no significant difference between two groups (P>0.05). Correlation analysis showed that FeNO and the declined rate of FeNO was negatively correlated with the ACT score(r=-0.302,P<0.05;r=0.674,P<0.01) and positively correlated with the improvement of ACT score (r=0.514,P<0.01;r=0.674,P<0.01). No significant correlation was found between FeNO and FEV1 or FEV1%pred. ConclusionThe effect of ICS/LABA therapy is better for asthma patients with higher FeNO. FeNO can be used for predicting the response to ICS/LABA therapy in patients with asthma and guiding the treatment.
Objective To investigate the effect of inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine on pancreas islets cultured with cytokines TNF-α and IL-1β in rats. Methods Islets isolated from Wistar rats were purified and cultured. According to whether cytokines TNF-α, IL-1β and aminoguanidine were added into the medium respectively or not, islets were divided into 4 groups: cultured with islet only was taken as blank control group, cultured with TNF-α+IL-1β as cytokine group, cultured with aminoguanidine as aminoguanidine group, and cultured with TNF-α+IL-1β and aminoguanidine as aminoguanidine+cytokine group. NO level in culture medium and iNOS activity in islets tissue (Test Kit), apoptosis (TUNEL method) and viability of islets cell (acridine orange/ethidium bromide stain), and the function of islets (insulin release test) were measured. Results Compared with blank control group, the activity of iNOS in islet tissue and level of NO in culture medium increased, and the mass mortality and apoptosis appeared in islet cells, while insulin secretion decreased in cytokine group (P＜0.01). Compared with cytokine group, the activity of iNOS 〔（3.17±0.51） U/ml vs. （38.93±4.72） U/ml〕 and level of NO 〔（50.5±10.4） μmol/L vs. （313.0±35.4） μmol/L〕 decreased, the survival 〔（72.73±3.14）% vs. （57.07±5.07）%〕 increased and the apoptosis rate 〔（20.11±8.48）% vs. （41.17±6.87）%〕 decreased, the insulin secretion (secretion index: 3.50±0.27 vs. 1.96±0.19) improved； There were all significant differences in 2 groups (P＜0.01). Conclusion The iNOS inhibitor aminoguanidine could prevent the islet from the damage of iNOS/NO, alleviate the impairment of cytokines to islets, and ameliorate the survival and function of islets.
Objective To assess the variation and its significance of messenger ribonucleic acid(mRNA) expression of endothelial nitric oxide synthase (eNOS) in allografts of common carotid transplantation model in white rabbits. Methods To establish an animal model of common carotid transplantation in vivo, 30 rabbits were divided into four groups with random number table. Group A (n=3): autografts; group B (n=9): allografts with the least treated; group C (n=9): allografts treated by penicillin/streptomycin and preserved under room temperature; group D (n=9): allografts treated by penicillin/streptomycin and cryopreserved in liquid nitrogen. All the transplanted grafts were harvested 1-3 weeks later, then compared and evaluated the histomorphological variation and eNOS mRNA expression. Results The vascular structures of autografts in group A were kept approximately normal, only a few infiltration of inflammatory cells could be found. The structural variations of allografts in other trial groups behaved similarly as, intima proliferation in the 1st week, intima hyperplasia in the 2nd week, and both intima and media hypertrophy in the 3rd week. And also there seemed that luminal thrombosis could be found in all the allografts. Allografts in group B were destructed utmost the worst in all the groups. The expression of eNOS mRNA in allografts of group B was significantly less than that in other groups (Plt;0.05). Conclusion The down-regulation of eNOS mRNA expression might lead to intima hyperplasia and thrombosis of allografts.
Objective To compare the value of fractional exhaled nitric oxide ( FENO) measurement and leukotriene D4 bronchial provocation test ( LTD4-BPT) in diagnosis and evaluation of asthma. Methods 20 uncontrolled,22 partially controlled, 20 controlled asthmatics, and 21 normal subjects were enrolled in the study. Measurement of FENO was performed followed by LTD4-BPT. The distribution characteristics and relationship of both results were analyzed, and the diagnostic value was compared using receiver operation characteristic ( ROC) curve.Results FENO was above 25. 0ppb in 80. 7% of the asthmatics. The proportion of asthmatics with FENO between 26.0ppb and 49.0ppb was larger in the uncontrolled and partially controlled subjects than that in the controlled subjects. Both the median and interquartile range of cumulative dosage ( PD20FEV1-LTD4) were much higher in the controlled asthmatics as compared with the uncontrolled and partially controlled asthmatics. The area under the ROC curve ( AUC) for PD20FEV1-LTD4 [ AUC: 0.914, 95% CI: ( 0.855, 0.974) ] was larger than that of FENO [ AUC: 0.820, 95% CI: ( 0.718, 0.921) ] . Higher sensitivity ( 0.8570 vs. 0.8065) and specificity ( 0.9048 vs. 0.7619) were in favor of PD20 FEV1 -LTD4 ≤ 4.800 nmol as compared with FENO ≥ 26.0ppb being the positive threshold. Conclusion Compared with FENO measurement, LTD4-BPT has higher sensitivity and specificity and is of higher diagnostic value for asthma.
Medical nitric oxide (NO) flow control system plays an important role in lowering pulmonary hypertension. The design requirements, overall scheme, delivery system and hardware circuits of a medical NO flow control system were introduced in this paper. Particularly, we proposed the design of NO delivery system and hardware circuits in detail. To deliver nitric oxide of a variable concentration, the designed system needs to work with a ventilator. The system can adjust and monitor the inhaled nitric oxide concentrations and send out sound and light alarms when the inhaled nitric oxide concentrations are out of the set range. To validate reliability and efficacy, we measured specifications such as linearity, stability and response time of the proposed NO flow control system by continuously administering nitric oxide into inspiratory circuit to deliver nitric oxide of variable concentrations to a test lung. The experiments showed that these specifications can meet the desired requirements.
【Abstract】Objective To investigate the expression of inducible nitric oxide synthase (iNOS) and p53 protein in hepatocellular carcinoma (HCC) and their relationship with angiogenesis. Methods Immunohistochemical method and image analysis technique were used to detect the expression of iNOS and p53 protein in tumor tissue sections of 59 HCC patients. Microvessel density (MVD) was counted by immunohistochemical staining with anti-CD34 antibody.Results ①The expression rates of iNOS and p53 were 81.4%(48/59), 64.4%(38/59) in HCC patients, respectively. The expression intensities of iNOS and p53 were 5 635±1 287, 3 352±873 in HCC patients, respectively. ②MVD was 32.5±2.73 in the tumor tissue of HCC patients. ③The expression of iNOS was correlated with the expression of p53 and MVD in HCC patients (P＜0.05); The expression of p53 was also correlated with the MVD in HCC patients (P＜0.05). Conclusion iNOS and p53 are highly expressed in HCC and may play a key role in angiogenesis of HCC.
【Abstract】ObjectiveTo investigate the protective effect of melatonin on renal injury induced by bile duct ligation in rats. MethodsSixtyfour rats were randomly divided into four experimental groups (n=16 rats per group): the control group (CN), sham operative group (SO), bile duct ligation group (BDL) and bile duct ligation melatonin treatment group (BDL+Mel). Obstructive jaundice was induced by common bile duct ligation. Plasma level of nitric oxide (NO), total bilirubin (TB), direct bilirubin (DB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and creatinine (Cr) were measured 4 d and 8 d after operation. NO and inducible nitric oxide synthase (iNOS) in renal tissue were detected at the same time point, too. Histopathological changes of kidneys were examined by HE staining. ResultsIn BDL group, the plasma levels of NO, TB, DB, ALT, AST, BUN and Cr were higher than those of SO group (P＜0.01）, and the level of NO and activities of iNOS in renal tissue were significantly increased (P＜0.01）. However, in BDL+Mel group, the plasma levels of NO, ALT, AST, BUN and Cr were lower than those of the BDL group (P＜0.01）, and the level of NO and activities of iNOS in renal tissue were significantly suppressed (P＜0.01）; histopathological changes of kidneys were improved.ConclusionAugmentation of iNOS activities and increasing of NO production in local tissue contributed to renal injury induced by bile duct ligation, and the mode of melatonin’s protective actions, at least in part, relates to interference with no pathways.
Objective To investigate the clinical characteristics of allergic and non-allergic asthma in Chinese adult asthmatic patients. Methods Consecutive treatment-naive adult outpatients with asthma were retrospectively analyzed in West China Hospital, Sichuan University from October 2014 to June 2016. The patients were classified into a non-allergic asthma (NAAS) group and an allergic asthma (AAS) group by skin prick test or antigen-specific IgE test. The differences between allergic and non-allergic asthma were compared in respect of gender, age, asthma control test (ACT) score, lung function, fractional exhaled nitric oxide (FeNO) level, body mass index (BMI), disease severity,etc. Results A total of 131 patients were enrolled in which 72 cases (54.96%) were allergic asthmatics and 59 cases (45.04%) were non-allergic asthmatics. The level of FeNO was statistically different (t=–2.762,P=0.007) between the NAAS group and the AAS group [(51.1±32.6)ppbvs. (69.1±41.7)ppb]. Seventeen cases of the NAAS group and 48 cases of the AAS group were complicated with rhinitis with statistically significant difference (χ2 =19.396,P=0.000). Airway limitation reversibility test showed that there were 37 cases in AAS and 20 cases in NAAS with no airway obstruction (NAO), 26 cases in AAS and 22 cases in NAAS with reversible airflow obstruction (RAO), 9 cases in AAS and 17 cases in NAAS with irreversible airflow obstruction (IAO), respectively, with statistically significant difference between two groups (Z=–2.461,P=0.014). There were 20 cases (33.9%) in NAAS and 37 cases (51.4%) in AAS with mild intermittent or persistent asthma, 18 cases (30.5%) in NAAS and 19 cases (26.4%) in AAS with moderate persistent asthma, 21 cases (35.6%) in NAAS and 16 (22.2%) in AAS with severe persistent asthma, respectively, with statistically significant difference (Z=–2.115,P=–0.034). The age, ACT score, FEV1%pred, and BMI had no statistical difference between two groups (allP>0.05). Conclusion Compared with allergic asthma, non-allergic asthma has less rhinitis, lower FeNO levels and higher prevalence of irreversible airflow obstruction.
ObjectiveTo evaluates the values of fractional exhaled nitric oxide (FENO) in the treatment of chronic cough prospectively.MethodsSubjects with chronic cough were recruited from the outpatient clinic of China-Japan Friendship Hospital. All the patients accepted FENO tests, sputum cell counts, pulmonary function tests, bronchial provocation tests, serum IgE, cough symptom scores and Leicester Cough Questionnaire before and after treatment of 4 weeks.ResultsThere were 29 patients with cough variant asthma (CVA), 19 patients with eosinophilic bronchitis (EB) and 39 patients with other causes. The baseline FENO level of the subjects whose coughs were relieved after inhaled corticosteroids (ICS) therapy of 4 weeks was (63±42) ppb, significantly higher than those with bad-response [(28±13) ppb, P<0.01]. The proportion of FENO decrease after ICS therapy was not only significantly related to the proportion of eosinophilic decrease (r=0.54, P<0.01), but also significantly related to the proportion of decrease of cough symptom scores (r=0.48, P<0.01). To distinguish the good responders from bad responders, the optimal baseline FENO cutoff value was 36 ppb, with sensitivity of 82%, specificity of 93%, positive predictive value of 94%, negative predictive value of 87%, accuracy of 83%.ConclusionsThere is a good relationship between the FENO decreasing levels after ICS therapy and the reliefs of cough symptoms in the CVA and EB patients. Chronic cough patients with FENO value more than 36 ppb are indicated to respond to ICS therapy.