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.
Objective To analyze the curative effect of nitric oxide (NO) and bosentan on treatment of the interruption of aortic arch (IAA) with ventricular septal defect (VSD) and serious pulmonary hypertension (SPH). Methods Thirty-two children with IAA and VSD combined SPH from January 2015 to May 2017 confirmed by cardiac CT and ultrasound in Children’s Hospital of Hebei Province were enrolled including 17 males and 15 females, aged 1.10-4.30 months (mean, 2.71±0.98 months) and weighing 3.33-6.10 kg (mean, 4.57±0.88 kg). The 32 children were randomly divided into two groups (n=16 in each), a NO group and a bosentan group. All the patients underwent interruption of aortic arch and ventricular septal defect repair. When patients returned to cardiosurgery intensive care unit (CSICU) half an hour later, patients in the NO group inhaled NO 20 ppm for 36 h and those in the bosentan group were given bosentan by nasogastric feeding 15 mg, twice a day. The cardic index, pulmonary/systemic pressure ratio, oxygenation index at 3 h, 6 h, 12 h, 24 h, 36 h after surgery were evaluated, and the differences between the two groups were compared. Results The pulmonary/systemic pressure ratio in the two groups increased at first and then decreased, while oxygenation index in the two groups decreased at first and then increased, and the differences in the same groups at the adjacent time points were statistically significant (P<0.05). The cardiac index in the two groups decreased at first and then increased, the differences in the same groups at the adjacent time points were statistically significant, except for 6 h and 12 h after surgery in the bosentan group (P>0.05). At postoperative 6 h, 12 h, the oxygenation index in the NO group was significantly higher than that in the bosentan group, and the pulmonary/systemic pressure ratio in the NO group was less than that in the bosentan group (P<0.01). The cardiac index in the NO group was higher than that of the bosentan group after 6 h, 12 h, 24 h of operation, which were statistically significant (P<0.05), and the cardic index of children in the NO group was greatly higher than that in the bosentan group after 12 h of surgery (P<0.01); at the same time point, the corresponding indexes were not statistically significant between the two groups (P>0.05). Conclusion NO inhalation in the treatment of IAA with VSD and SPH in children with early postoperative SPH is better than the bosentan, but in the late postoperative period, the effect is similar.
【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.
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.
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.
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.
ObjectiveThrough measuring fractional exhaled nitric oxide (FeNO) and eosinophil levels of peripheral blood in chronic obstructive pulmonary disease (COPD) patients with different phenotype of acute exacerbation frequency, to predict the therapeutic effect of glucocorticoid therapy and guide the clinical treatment of different subtypes patients with acute exacerbations of COPD.MethodsA total of 127 patients with acute exacerbation of COPD in Suining Central Hospital from February 2017 to October 2019 were recruited. They were divided four groups according to the number of acute exacerbations in the past one year and the treatment scheme, ie. a frequent acute exacerbation with glucocorticoid treatment group (34 cases), a frequent acute exacerbation with non-glucocorticoid treatment group (31 cases), a non-frequent acute exacerbation with glucocorticoid treatment group (30 cases), and a non-frequent acute exacerbation with non-glucocorticoid treatment group (32 cases). FeNO value, eosinophil ratio in peripheral blood, COPD assessment test (CAT) score, and interleukin-8 (IL-8) concentration were measured before and on the 10th day of treatment, and the differences within group and between groups before and after treatment were compared.ResultsCAT score, FeNO, eosinophil ratio and IL-8 level in the four groups were significantly improved on the 10th day after treatment (all P<0.05). The declines of FeNO value, eosinophil ratio, and IL-8 level on the 10th day of treatment compared with those before treatment in the frequent acute exacerbation with glucocorticoid treatment group and the frequent acute exacerbations with non-glucocorticoid treatment group were larger than those in the non-frequent acute exacerbation with glucocorticoid treatment group and the non-frequent acute exacerbation with non-glucocorticoid treatment group (all P<0.05). The declines of FeNO value, blood eosinophil ratio and IL-8 level in the frequent acute exacerbation with glucocorticoid treatment group were also statistically significantly larger than those in the frequent acute exacerbations with non-glucocorticoid treatment group (all P<0.05). The improvement of CAT score in the frequent acute exacerbation with glucocorticoid treatment group was greater than that in other three groups (all P<0.05). There was no significant difference in CAT score between the non-frequent acute exacerbation with glucocorticoid treatment group and the non-frequent acute exacerbation with non-glucocorticoid treatment group (P>0.05).ConclusionsThe degree of airway inflammation is more obvious in patients with frequent acute exacerbation phenotype of COPD. FeNO value can reflect the level of airway inflammation in patients with frequent acute exacerbation of COPD and evaluate the response to glucocorticoid therapy.
Objective To investigate the efficacy of low-dose inhaled nitric oxide (iNO) in the treatment of severe hypoxemia after Sun’s operation. Methods The clinical data of patients undergoing Sun’s operation for acute Type A aortic dissection in our hospital from January 2020 to June 2022 were retrospectively analyzed. Patients who received conventional treatment before November 2021 were enrolled as a control group. After November 2021, iNO was used in our hospital, and the patients who received iNO as an iNO group. The preoperative clinical baseline data, perioperative clinical data and oxygenation index were compared between the two groups. Results A total of 54 patients were included in the control group, including 45 males and 9 females, with an average age of 53.0±10.9 years. A total of 27 patients were included in the iNO group, including 21 males and 6 females, with an average age of 52.0±10.6 years. The preoperative body mass index of the two groups was greater than 25 kg/m2, white blood cell count, C-reactive protein were significantly higher than normal level, but there was no statistical difference between the groups (P＞0.05). There were no statistical differences in intraoperative data between the two groups (P＞0.05). The iNO group had significantly shorter duration of mechanical ventilation, postoperative ICU stay, and postoperative hospital stay than the control group (P<0.001). After 12 h of iNO treatment, hypoxic condition improved obviously, oxygenation indices in 12 h, 24 h, 36 h,48 h, 60 h and 72 h in the iNO group were significantly higher than those in the control group (P<0.05). Conclusion The treatment of severe hypoxemia after Sun’s surgery with low-dose of iNO is safe and effective, can significantly improve oxygenation function, and has significant advantages in shortening ventilator use time, postoperative ICU stay and postoperative hospital stay, but it is not significant in changing postoperative mortality.
Objective To investigate the relationship between delayed diagnosis time (time from symptom onset to diagnosis) in patients with chronic obstructive pulmonary disease (COPD) and the burden of type 2 inflammation (defined as the persistent inflammatory status assessed by blood EOS counts, EOS%, and Fractional exhaled nitric oxide(FeNO) among other biomarkers).MethodsThis study was a single-center, observational study that included patients with COPD first diagnosis at the respiratory outpatient department of our hospital from June 2023 to December 2024. Asthma-COPD overlap (ACO) were identified according to the 2017 Spanish COPD guidelines. Clinical data were collected, including gender, age, delayed diagnosis time, acute exacerbations in the past year, pulmonary function tests, exhaled nitric oxide (FeNO), and type 2 inflammatory markers such as blood eosinophil counts (EOS). The correlation between the delayed diagnosis time and type 2 inflammation burden, as well as its influencing factors, were analyzed. Results A total of 195 patients were included, with 98 cases of COPD and 97 cases of ACO. The mean delayed diagnosis time was 18.0 (2.8, 37.5) months for the overall patients, 24.0 (1.0, 60.0) months for COPD, and 16.5 (3.0, 36.0) months for ACO, with no significant difference between the COPD and ACO groups (P>0.05). The median blood EOS counts, EOS%, andFeNO levels were 180 cells/μL, 1.9%, and 18 ppb in the COPD group, respectively, compared to 350 cells/μL, 4.7%, and 28 ppb in the ACO group, indicating higher type 2 inflammation levels in the ACO group (all P<0.001). A significant correlations were found between the disease course and the blood EOS counts and EOS% of the patients (respectively r=0.159, 0.152, all P<0.05).FeNO levels showed no significant correlation with delayed diagnosis time of COPD (P>0.05). Patients with a history of asthma and acute exacerbations in the past year had longer delayed diagnosis time and higher peripheral blood eosinophil counts (all P<0.05). Binary logistic regression analysis revealed that BMI and delayed diagnosis time were independent influencing factors for blood EOS counts (all P<0.05). ConclusionDelayed diagnosis of COPD was associated with aggravated type 2 inflammatory burden. Clinical practice should emphasize early recognition of COPD symptoms and implement prompt therapeutic interventions.
Objective To study the effects of L-arginine （L-Arg） on cell proliferation, inducible nitric oxide synthase (iNOS) expression and cell cycle in human colon carcinoma cell line LS174 through nitric oxide (NO) pathway. Methods LS174 cells were cultured in medium with L-Arg at different concentrations for different times. MTT method was employed to evaluate the level of the cell proliferation. The production of NO in culture supernatants of LS174 cell was detected with enzyme reduction of nitrate. The distribution of the cell cycle was detected with the flow cytometry (FCM). The expression level of iNOS in the cells was determined by Western blot and SP immunocytochemical staining method. Results The growth of LS174 was promoted by the L-Arg at low concentration (0.125 mmol/L) and inhibited at high concentrations (0.5， 2， 8 and 32 mmol/L). The level of NO was increased with the increasing concentration of L-Arg in culture medium. To compare with the control group, the ratio of cells at S phase was increased after 48 hours’ treatments with high concentrations (0.5, 2, 8 and 32 mmol/L) of L-Arg (P＜0.05, P＜0.01); while there was no obvious difference after treatments with low concentration (0.125 mmol/L) of L-Arg (Pgt;0.05). With the increase of the concentration of L-Arg, the expression of iNOS was increased as compared with control group. The higher the concentration of L-Arg was, the better the effect. Conclusion L-Arg can induce the expression of iNOS resulting in increase the production of nitric oxide (NO). Low concentration of L-Arg can promote the growth of LS174 cells, while high concentration ones can inhibit growth and proliferation. The high concentration of L-Arg could induce S phase arrestion in the cell cycle.