Changes of hydrogen sulfide and heme oxygenase-1 in blood of children with congenital heart disease and pulmonary hypertension before and after surgery

AIM: To evaluate the changes and clinical significance of hydrogen sulfide (H2S) and heme oxygenase-1 (HO-1) in patients with congenital heart disease (CHD) and pulmonary hypertension (PH). METHODS: Forty-eight patients with CHD and left-to-right shunt were selected randomly from cardiac surgery in the Fourth Hospital of Hebei Medical University. Forty-eight patients were divided into 3 groups according to the pulmonary artery systolic pressure (PASP) by Doppler echocardiography before operation: group A was normal (PASP<30 mmHg, n=15); group B was mild PH (PASP 30-49 mmHg, n=15); group C was moderate-severe PH (PASP≥50 mmHg, n=18). The radical operation was performed successfully under complex intravenous anesthesia in all 48 patients. The radial arterial blood was collected immediately before operation, 1 h and 24 h after operation. The concentration of plasma H2S was measured by optical absorbance method at 670 nm and HO-1 activity in serum was detected by dual wavelength spectrophotometer at different time points. RESULTS: Plasma level of H2S in all groups after operation was higher than that in the same group before operation. Significant difference of the plasma H2S level was observed between pre-operation and 24 h post-operation in the same group (P<0.05). The plasma level of H2S in group C and group B at time points of pre-operation, 1 h post-operation or 24 h post-operation was significantly lower than that in group A, the plasma level of H2S in group C was significantly lower than that in group B. Compared to the values in each group before operation, the activity of serum HO-1 among groups after operation was not obviously and statistically different (P>0.05). No obvious difference of HO-1 activity in blood among 3 groups was observed (P>0.05). There was a negative correlation between PASP and the level of H2S in 3 groups at time points of pre-operation, 1 h and 24 h post-operation (pre-operation, r=-0.66, P<0.01; 1 h post-operation, r=-0.458, P<0.01; 24 h post-operation, r=-0.730, P<0.01). No correlation between PASP and HO-1 activity in blood serum was found. CONCLUSION: H2S plays an important role in the formation of PH and remodeling of pulmonary vasculature. There is no correlation between PASP and HO-1 activities in blood, but HO-1 may play an indirect action in the formation of PH and remodeling of pulmonary vasculature. Measuring the level of H2S may be a reliable method to follow the change of pulmonary pressure and worsened PH.     

Change of intestinal flora and its relationship with IL-23/IL-17 axis in patients with ulcerative colitis

AIM:To investigate the change of intestinal flora distribution and its relationship with interleukin-23 (IL-23)/IL-17 axis in ulcerative colitis (UC) patients. METHODS:The fresh fecal samples from 20 patients with active UC and 20 healthy controls were collected. The distribution of the flora was analyzed by direct smear and traditional bacterial culture. The changes of bacteria were detected by real-time PCR. The hemoglobin, albumin, erythrocyte sedimentation, and C-reactive protein levels were tested routinely. Both normal and damaged mucosal tissues of UC patients were examined and obtained by colonoscopy, and further assessed by Mayo scoring, Baron grading and HE staining. The expression of IL-17 and IL-23 was observed by immunohistochemistry and Western blot. RESULTS:(1) The degree of flora imbalance in active UC patients was higher than that in the healthy controls (P<0.05). (2) The results of aerobic culture showed that the number of Escherichia coli in the UC patients was significantly lower than that in the normal controls (P<0.01), while Enterococcus was increased obviously (P<0.01). The results of anaerobic culture revealed that the numbers of Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients were significantly decreased (P<0.01). (3) Quantitative analysis of target bacteria showed that the relative quantification of Escherichia coli, Bacteroidetes, Bifidobacterium bifidum and Lactobacilli in the UC patients was significantly lower than that in the normal subjects, and the number of Enterococcus was significantly increased (P<0.01). (4) Compared with control group, no significant change of hemoglobin in the UC patients was ovserved, albumin was significantly decreased (P<0.05), but erythrocyte sedimentation and C-reactive protein levels were elevated obviously (P<0.01). (5) The Mayo score, Baron grade, and histopathological score were all increased (P<0.01). (6) High IL-17 and IL-23 expression levels were detected in the UC patients (P<0.01). (7) Correlation analysis showed that the average absorbance values of IL-17 and IL-23 expression were positively correlated with Baron grade (r=0.717, P=0.02; r=0.849, P=0.016) and pathological score (r=0.660, P=0.03; r=0.675, P=0.032). Meanwhile, the average absorbance value of IL-23 expression was negatively correlated with the number of Escherichia coli (r=-0.699, P=0.025), and positively correlated with Enterococcus (r=0.872, P=0.010). Furthermore, the average absorbance value of IL-17 expression was positively correlated with Enterococcus (r=0.764, P=0.046), and both of them were not correlated with other bacteria. CONCLUSION:Obvious flora imbalance exists in active UC patients, changed intestinal microflora is closely related with the degree of inflammation. IL-23/IL-17 axis, as a key factor in the development of UC, may be related to the changes of intestinal microflora. The interaction between intestinal microflora and IL-23/IL-17 axis plays an important role in the pathogenesis of UC.     

Changes in endothelin-1 and atrial natriuretic factor and the effect of atrial natriuretic factor on the change of endothelin-1 in asthmatic guinea pigs

AIM: To investigate the possible role of endothelin(ET-1) in asthma pathogenesis and the effect of atrial natriuretic factor (ANF) on changes of ET-1. METHODS: Measuring the contents of endothelin-1(ET-1),atrial natriuretic factor(ANF),cGMP in plasma and bronchoalveolar lavage fluid (BALF) of guinea pigs. RESULTS: The contents of ET-1, ANF and cGMP in asthma group were higher than that of the control group; There was a significant negative correlation between ET-1 and ANF( r=-0.638,P <0.05) in plasma of the asthma group, and a significantly negative correlation between ET-1 and ANF( r=-0.921,P <0.01) in BALF of the asthma group. There was a significantly positive correlation between ANF and cGMP( r=0.848,P <0.01) in plasma of the asthma group,and a significantly positive correlation between ANF and cGMP ( r=0.831,P <0.01) in BALF of the asthma group. The levels ET-1 in the asthma+rat ANF(rANF) group were lower than those in the asthma group,and the levels of cGMP in the asthma+rat ANF(rANF) group were higher than those in the asthma group after ceasing to infuse rANF for guinea pigs for 30 minutes.CONCLUSION: ET-1 might play an important role in the pathogenesis of asthma.ANF might inhibit production of ET-1.     

Effect of Herba Taxilli extracts combined with miR-375 on viability and apoptosis of osteoarthritic chondrocytes

AIM To study the effects of extracts of Herba Taxilli (Sangjisheng, SJS) on the viability and apoptosis of osteoarthritic chondrocytes and the underlying mechanism. METHODS Human primary osteoarticular chondrocytes (RPOC) were divided into control group, interleukin-1β (IL-1β) group, IL-1β+low-dose extracts of SJS (SJS-L) group, IL-1β+medium-dose extracts of SJS (SJS-M) group, IL-1β+high-dose extracts of SJS (SJS-H) group, IL-1β+anti-miR-NC group, IL-1β+anti-miR-375 group, IL-1β+SJS-H+miR-NC group, IL-1β+SJS-H+miR-375 group. The cell viability was measured by MTT assay, apoptosis was analyzed by flow cytometry, miR-375 expression was detected by qPCR, and the protein levels of cyclin D1, P21, Bcl-2, Bax and caspase-3 were determined by Western blot. RESULTS Compared with control group, the viability of RPOC at 24 h, 48 h and 72 h and the protein expression levels of cyclin D1 and Bcl-2 were significantly decreased (P<0.05), the protein levels of P21, Bax and caspase-3, the apoptotic rate and the expression level of miR-375 were remarkably increased in IL-1β group(P<0.05). Compared with IL-1β group, the cell viability at 24 h, 48 h and 72 h and the protein expression of cyclin D1 in the RPOC were greatly increased (P<0.05), while the expression of P21 was significantly decreased in IL-1β+SJS-M group and IL-1β+SJS-H group(P<0.05).The apoptotic rate, Bax, caspase-3 protein and miR-375 expression were obviously decreased (P<0.05), and Bcl-2 protein level was significantly increased in IL-1β+SJS-H group compared with IL-1β group(P<0.05). Compared with IL-1β+anti-miR-NC group, the expression of miR-375, the protein levels of P21, Bax, caspase-3 and the apoptotic rate in the RPOC of IL-1β+anti-miR-375 group were markedly decreased (P<0.05), while the cell viability at 24 h, 48 h and 72 h and the protein levels of cyclin D1 and Bcl-2 were significantly increased (P<0.05). Over-expression of miR-375 reversed the effects of extracts of SJS on the viability and apoptosis of RPOC with IL-1β stimulation. CONCLUSION The extracts of Herba Taxilli promotes the viability and inhibits apoptosis of RPOC treated with IL-1β, which is related to the regulation of miR-375 expression.     

Changes and clinical implications of plasma ANP,BNP and CNP levels in type 2 diabetic patients with vascular complications

AIM:To examine the changes and clinical implications of the plasma atrial natriuretic peptide (ANP), brain natriuretic peptide(BNP) and C-type natriuretic peptide(CNP) levels in type 2 diabetic patients with vascular complications. METHODS:9 subjects without diabetes, 34 diabetic patients without vascular complications and 23 diabetic patients with vascular complications were enrolled in the study and categorized into three groups. Plasma proANP, BNP fragment and N-terminal pro-CNP (NT-proCNP) were measured by enzyme linked immunosorbent assay (ELISA). The changes and associations of the plasma natriuretic peptide levels among three groups were analyzed. RESULTS:Compared with those in the controls and diabetics without vascular complications, the plasma proANP and BNP fragment levels in diabetic patients with vascular complications increased significantly (P<0.01). However, the plasma NT-proCNP level in this group decreased significantly (P<0.01). Nevertheless, no significant differences in the plasma proANP, BNP fragment and NT-proCNP levels among three subgroups (diabetics patients with macrovascular and/or microvascular complications) were observed (P>0.05). The plasma ANP level of diabetic patients with vascular complications was corelated significantly with the BNP levels (r=0.309, P<0.05). Furthermore, the ANP and BNP levels were correlated significantly with the CNP level (r=-0.374, P<0.05; r=-0.653, P<0.01). CONCLUSION:Measurement of the plasma ANP, BNP and CNP levels together may be a simple, cost-effective and reliable way to screen for diabetic patients with vascular complications.     

Effects of combination treatment with perindopril and losartan on left ventricular remodeling and cardiac function in patients with myocardial infarction

AIM: Inhibiting the renin-angiotensin-aldosterone system prevents left ventricular (LV) remodeling after myocardial infarction (MI). This study was designed to assess the effects of a combination of perindopril and losartan on LV remodeling, cardiac function and serum procollagen type Ⅲ aminoterminal peptide (PⅢNP) levels in patients with acute MI.METHODS: Patients with anterior MI were divided into 3 groups: MI+perindopril, MI+losartan, and MI+perindopril+losartan. After successful intervention therapy, perindopril 2-4 mg/d or losartan potassium 25-50 mg/d or combination of the both were administered. All patients took aspirin, clopidogrel and statins, while some of the patients were treated with beta-blockers, nitrate and a platelet glycoprotein IIb/Ⅲa receptor antagonist. Three months later, LV dimensions and LV ejection fraction (LVEF) were measured by ultrasonography. Plasma brain natriuretic peptide (BNP), serum C-reactive protein (CRP) and PⅢNP levels were evaluated with ELISA or RIA.RESULTS: Baseline characteristics of the 3 groups were the same. All patients showed decreased CRP, increased BNP and PⅢNP levels, and LV dilation and dysfunction after treatment for three months. Compared with the 2 single therapy groups, patients in the combination group showed significantly lower CRP, BNP and PⅢNP levels, less LV dilation and higher LVEF. Serum PⅢNP level was positively correlated with CRP level and LV end-diastolic volume index(r=0.597 and r=0.543, respectively,both P<0.01), and negatively correlated with LVEF(r=-0.565, P<0.01).CONCLUSION: For patients with AMI, combination of perindopril and losartan significantly inhibited LV remodeling and improved LV function. Inhibition of myocardial interstitial fibrosis might be part of the mechanism.     

Levels of CGRP and ET-1 in plasma of pulmonary artery and thoracic aorta and in the extractives of lung and ventricular tissues of the chronic hypoxic rats

AIM AND METHODS: To explore the effects calcitonin gene-related peptide (CGRP) and endothelin-1(ET-1) on the mechanisms of hypoxic pulmonary hypertension (HPH), the contents of CGRP and ET-1 in plasma of pulmonary artery and thoracic aorta and in extractives of lung and ventricular tissues of the chronic hypoxic rats were determined by radioimmunoassay. The changes of their hemodynamic indices and right heart hypertrophy index were monitored simultaneously. RESULTS: The level of pulmonary artery plasma CGRP was significantly higher than that of thoracic aorta plasma, but just the reverse was ET-1 or the ratio of ET-1 and CGRP in control rats( P<0.01). Compared with controls, the level of pulmonary artery plasma CGRP was gradually reduced in all hypoxic rats, but ET-1 was enhanced after 7 and 14 days of hypoxia and was decreased after 21 days of hypoxia. With prolonging time exposed to hypoxia, the level of thoracic aorta plasma CGRP was markedly increased in hypoxic animals compared with controls ( P<0.05), the positive correlation significantly with increased pulmonary arterial pressure( r = -0.896, P =0.05), but the lower level of thoracic aorta plasma ET-1 showed or negative correlation with pulmonary arterial pressure. CONCLUSIONS: These data suggest that the unbalance of effects of CGRP and ET may plays an important role in regulating the resistance of pulmonary circulation and has close relation with the formation of HPH.     

Effects of Xinkang recipe on myocardial miR-25-3p expression and SERCA2a activity in heart failure rats

AIM: To investigate the effects of Xinkang recipe on myocardial miR-25-3p expression and sarcoplasmic reticulum calcium ATPase 2a (SERCA2a) activity in heart failure rats. METHODS: Male SD rats were randomly divided into normal group, sham group, model group, Xinkang recipe group (Xinkang group), and captopril group. The heart failure rat model was induced by intraperitoneal injection of doxorubicin. Distilled water, Xinkang recipe and captopril were administrated by gastric gavage for 35 d, respectively. The indexes of cardiac function and plasma level of brain natriuretic peptide (BNP) were measured. The SERCA2a activity was determined by the inorganic phosphorus method. The myocardial protein expression of SERCA2a and phospholamban (PLB) was detected by Western blot. The myocardial expression of miR-25-3p was detected by stem-loop RT-qPCR. RESULTS: Cardiac output (CO), left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) in Xinkang group and captopril group were significantly higher while the plasma levels of BNP were significantly lower than those in model group (P<0.01). The myocardial expression levels of miR-25-3p in Xinkang group and captopril group were significantly lower while the myocardial protein le-vels of SERCA2a and PLB were significantly higher than those in model group (P<0.01). The SERCA2a/PLB ratio and SERCA2a activity in Xinkang group were significantly higher than those in model group (P<0.05), and no significant change was observed between captopril group and model group. CONCLUSION: Xinkang recipe therapy may improve cardiac function in heart failure rats, which may be related to inhibiting the expression of miR-25-3p, increasing the SERCA2a/PLB ratio and enhancing SERCA2a activity in the myocardium.     

Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

Relationship between Th1/Th2 balance and CD28/CTLA-4 expression on peripheral blood T cells in patients with systemic lupus erythematosus

AIM: To investigate the pattern of Th1/Th2 balance in systemic lupus erythematosus(SLE) patients and the relationship between CD28/CTLA-4(cytotoxic T-lymphocyte antigen-4) molecule expression and Th1/Th2 balance.METHODS: Eighteen SLE patients met the ARA 1997 updated SLE criteria were selected in the study. According to Bombardier's SLEDAI criteria, all patients were classified into two groups: active group(12 cases) and static group(6 cases). Fourteen normal individuals, matched for age and sex of the patients, served as controls. The peripheral blood mononuclear cells(PBMCs) were isolated by density gradient centrifugation and cultured in RPMI-1640 culture medium. After treated with PMA(5 μg/L) and ionomycin(500 μg/L) for 72 h, the PBMCs were collected, the contents of IFN-γ and IL-10 in the supernatant of cultured PBMCs were detected using enzyme linked immunosorbent assay(ELISA). The expression of CD28 and CTLA-4 molecules on T cells were detected by flow cytometric technique with double staining by FITC or PE labeled monoclonal antibodies. RESULTS: The level of IL-10 was higher in the PBMCs of active and static SLE patients(351.29 ng/L±153.31 ng/L and 319.37 ng/L±153.39 ng/L) than that in controls(254.48 ng/L±120.69 ng/L), but the difference did not reach statistical significance(P>0.05). The level of IFN-γ was significantly lower in the PBMCs of active SLE patients(25.76 ng/L±16.09 ng/L) than that in controls(50.71 ng/L±27.92 ng/L, P＜0.05). The ratio of IL-10/IFN-γ was significantly higher in active SLE patients(18.74±13.77) than that in controls(6.66±4.95, P<0.05). Either before or after culture, the expression of CD28 molecule on CD3⁺and CD8⁺ T cells from all SLE patients was not remarkably different from that in the cells of controls. Before culture, the expression of CTLA-4 molecule on CD3⁺T cells of active SLE patients(0.79%+0.37%) was significantly lower than that in the cells of controls(1.31%+0.61%, P<0.05). After culture, the expression of CTLA-4 molecule on CD3⁺ T cells of SLE patients was still lower than that in the cells of normal controls without statistical significance(P＞0.05).The expression level of CD28 molecule on CD3⁺ or CD8⁺ T cells in active SLE patients and controls was not correlated with the levels of IFN-γ and IL-10 in the supernatants(P＞0.05). The level of CTLA-4 molecule expression on CD3⁺ T cells of active SLE patients was positively correlated with IFN-γ level(r=0.681, P<0.05), while was negatively correlated with IL-10 levels(r=-0.624,P＜0.05) and the ratio of IL-10/IFN-γ(r=-0.738, P<0.01). The level of CTLA-4 molecule expression on CD3⁺ or CD8⁺ T cells of controls showed no correlation with IFN-γ levels, while showed negative correlations with IL-10 level(r=-0.587, P<0.05; r=-0.563, P<0.05, respectively).CONCLUSION: There is a bias in the differentiation of Th0 cells towards Th2 in SLE patients. CTLA-4 probably plays an important role in this mechanism through suppressing the signal transmitted by CD28.     

Relationship between the changes of angiotensin Ⅱ and vascular endothelial growth factor in plasma and the occurrence of diabetic nephropathy

AIM:To measure the concentrations of angiotensinⅡ(ATⅡ) and vascular endothelial growth factor(VEGF) in plasma of diabetics and investigate the relationship between ATⅡ and VEGF and the role of VEGF in occurrence and development of diabetic nephropathy(DN). METHODS:98 type Ⅱ diabetics were divided into two groups: 43 were in diabetic nephropathy group and 55 in non-diabetic nephropathy (NDN) group, and 25 healty persons were in control group. We measured the levels of ATⅡ, VEGF, glycosylated hemoglobinC (GHbAlc) in plasma and urinary microalbumin(UmALB), respectively.RESULTS:The levels of ATⅡ, VEGF, HbAlc and UmALB in DN group and NDN group were increased significantly compared with control group(P＜0.01); The concentrations of ATⅡ, VEGF and UmALB in DN group were obviously higher than that in NDN group(P＜0.01), but there was no significant difference of GHbAlc between DN and NDN; There was a positive correlation between ATⅡ and VEGF(r=0.465,P＜0.05), and VEGF was positively correlated with UmALB(r=0.540,P＜0.01). CONCLUSION:The levels of ATⅡand VEGF increased significantly in plasma of diabetics, and ATⅡ and VEGF may involve in the occurrence and development of diabetic nephropathy.     

Diagnostic value of Th17 cells in symptom severity and prognosis of chronic obstructive pulmonary disease

AIM: To observed the correlation between Th17 cell level and the symptom severity and prognostic factors of chronic obstructive pulmonary disease (COPD), and to explore the clinical application value of Th17 cell level in assessing the prognosis of patients with COPD. METHODS: The patients with diagnosed COPD (n=110) in our hospital during May 2013 to December 2014, and 40 healthy subjects were enrolled in the study. According to the Global Initiative for Chronic Obstructive Lung Disease (GOLD), the COPD patients were divided into group A (low risk, less symptoms), group B (low risk, more symptoms), group C (high risk, less symptoms) and group D (high risk, more symptoms), which were given inhaled corticosteroid/long-acting β₂ agonist or corticosteroid/long-acting β₂ agonist+long-acting antimuscarinic agent treatment for 3 months. The proportion of Th17 cells, cytokines (IL-17 and IL-6), the COPD assessment test (CAT) score, age, body mass index, pulmonary function and the times of acute exacerbation of COPD in previous 1 year were observed before and after treatment. The correlation analysis between the level of Th17 cells and other clinical characteristics was performed. RESULTS: Th17 cell, IL-17 and IL-6 levels in COPD group were significantly higher than those in control group (P < 0.05). With the increase in the severity of COPD symptoms, Th17 cells, cytokines (IL-17 and IL-6) and CAT score in groups B and D were significantly higher than those in groups A and C (P < 0.05). The univariate analysis showed that the levels of Th17 cells in groups B and D before treatment were positively correlated with the CAT score (P < 0.05), which were negatively correlated with FEV₁ , FEV₁% Pred, FVC and FVC% Pred. The levels of Th17 cells were not correlated with the CAT score, FEV₁, FEV₁% Pred, FVC and FVC% Pred in groups A and C. The levels of Th17 cells after treatment were positively correlated with the CAT score, which were negatively correlated with FEV₁ , FEV₁% Pred, FVC and FVC% Pred (P < 0.05). CONCLUSION: The peripheral Th17 cell level has a good correlation with IL-17, IL-6, CAT score and pulmonary function in COPD patients, suggesting a potential value to predict the symptom severity and prognosis of COPD.     

转哺乳动物cyp2e1基因矮牵牛耐甲醛胁迫的生理机制

细胞色素P450 CYP2E1酶主要存在于哺乳动物肝细胞中,在代谢异源有机物方面起着重要作用。前期研究发现,转cyp2e1矮牵牛显著提高了对甲醛的抗性。以转cyp2e1矮牵牛为试验材料,分析其对甲醛胁迫响应的相关生理指标。结果显示,在甲醛胁迫下,转cyp2e1矮牵牛细胞中的MDA含量低于转gus和野生型矮牵牛,SOD和POD活性均高于转gus和野生型,乙醇脱氢酶(ADH)活性稍有增强,且消耗更多的谷胱甘肽。此外,在甲醛胁迫下,转cyp2e1矮牵牛的IAA、ZRs和ABA含量呈现下降而GA含量呈现上升趋势;但转gus和野生型矮牵牛IAA、ZRs和ABA含量呈现上升而GA含量呈现下降趋势。转cyp2e1矮牵牛在含有50 mg·L-1甲醛的处理液中孵育72 h后,处理液中甲醛含量接近为0;而转gus和野生型处理液中仍有近50%的甲醛。     

Effect of non-invasive ischemic preconditioning on nitric oxide/endothelin-1 imbalance and gas exchange impairment following limb ischemia reperfusion: a clinical study

AIM: To investigate the effects of non-invasive ischemic preconditioning on nitric oxide (NO)/endothelin-1 (ET-1) imbalance and gas exchange impairment following limb ischemia reperfusion in patients undergoing unilateral lower extremity surgery with tourniquet. METHODS: Twenty-seven patients aged 25-65 years, whose tourniquets duration varied from 1 h to 1.5 h and matched American Society of Anesthesiologists Physical Status Ⅰ-Ⅱ, were randomized into two groups: a control group (n=14) and a ischemic preconditioning group (IPC group, n=13) in which patients received three cycles of 5 min of ischemia/5 min of reperfusion before tourniquet inflation. Radial arterial blood gas, plasma malondialdehyde (MDA) and NO, serum ET-1 and interleukin-6 (IL-6) were measured just before tourniquet inflation(T0), 1 h after inflation(T1), and 0.5 h(T2), 2 h(T3), 6 h(T4), 24 h(T5) after tourniquet deflation. Meanwhile NO/ET-1 ratio, alveolar-arterial oxygen gradient (PA-aDO2) and intrapulmonary shunt (Qs/Qt) were calculated. RESULTS: In control group, arterial partial pressure of oxygen (PaO2) were decreased, while PA-aDO2 and Qs/Qt were increased significantly at T4 compared to the baselines at T0 (P<0.01). Plasma NO levels and NO/ET-1 ratios decreased gradually after tourniquets deflation and statistical significances were observed at T3 (P<0.01) with a valley at T4 (P<0.01) and recovered to baselines at T5. Serum ET-1, IL-6 and plasma MDA began to increase remarkably after T3 (P<0.05 or P<0.01), peaked at T4 and dropped slightly at T5. The changes above-mentioned could be well attenuated by the application of IPC (P<0.05 or P<0.01) except PaO2 (P>0.05). CONCLUSION: Clinical application of unilateral tourniquet within safe time limit (1.5 h) may lead to limb ischemia reperfusion and further pulmonary gas exchange impairment, which could be partially attenuated by the application of IPC via alleviating NO/ET-1 imbalance.     

Subclasses of serum HDL in middle and old aged patients with hyperlipidemia in Chengdu City

AIM: To detect the change of composition and ratio of serum HDL subclasses and explore the relationship between these changes and the plasma lipid level in patients with hyperlipidemia. METHODS: The components of subclasses of serum HDL in 172 middle and old aged patients with hyperlipidemia and 115 healthy middle and old aged were determined by dimensional gel electrophoresis associated with immuno-blotting method. RESULTS: Compared to the healthy controls, the contents of pre β1-HDL，　HDL3b and HDL3a were significantly higher (P＜0.05 or P＜0.01), while that of HDL2b was significantly lower (P＜0.01) in middle and old aged patients with hyperlipidemia. The content of pre β1-HDL increased with age in healthy controls, whereas the HDL2b decreased. The content of pre β1-HDL was significantly higher (P＜0.05), while the HDL2b (P＜0.05) was significantly lower in men than in women in patients with hyperlipidemia and the healthy controls. In middle and old aged patients with hyperlipidemia, the content of pre β1-HDL was positively correlated with the serum TG, TC, apoB100, apoCⅡ, apoCⅢ, apoE and TG/HDL-C (r=0.432; r=0.243; r=0.341; r=0.259; r=0.335; r=0.308 and r=0.453, P＜0.05 or P＜0.01), while it was negatively correlated with HDL-C (r=－0.167, P＜0.05). The content of HDL2b was negatively correlated with TG, TC, apoCⅡ, apoCⅢ and TG/HDL-C (r=－0.296; r=－0.156; r=－0.182; r=－0.216; r=－0.203 and r=－0.313, P＜0.05 or P＜0.01), while it was positively correlated with HDL-C (r=0.124, P＜0.05). CONCLUSIONS: The particle of HDL in the middle and old aged patients with hyperlipidemia showed a general shift towards smaller size, which indicated that the reverse cholesterol transport might be weakened. Men had smaller HDL particle size than women.     

Effects of Tongxinluo on connexin 43 remodeling and ventricular arrhythmia after myocardial infarction in rats

AIM: To determine the effects of Tongxinluo(TXL) on connexin 43(Cx43) remodeling and ventricular arrhythmia(VA) after myocardial infarction(MI) in rats. METHODS: Male SD rats were randomly divided into sham-operated(sham) group(n=25) and operation group(n=75). The left anterior descending(LAD) was ligated in operated group, while the rats in sham group only underwent pericardiotomy. The rats in operation group which survived for 3 d after operation were randomly assigned to TXL group and MI group. The rats in TXL group was administrated with TXL(2 g·kg^-1·d^-1, intragastric administration) for 4 weeks, while normal saline was applied to the rats in sham group and MI group. The levels of interleukin-1β(IL-1β) and endothelin-1(ET-1) in the tissue from the border zone were measured by ELISA after treatment. The distribution and the mRNA and protein expression of Cx43 were detected by immunohistochemical staining, RT-PCR and Western blotting, respectively. The burst pacing was used to induce ventricular arrhythmia(VA). RESULTS: Compared with sham group, the levels of IL-1β and ET-1 and the incidence of VA were significantly increased, while the mRNA and protein expression of Cx43 was markedly reduced with irregular distribution in MI group(P<0.05). Compared with MI group, the levels of IL-1β and ET-1 and the incidence of VA were significantly reduced, while the expression of Cx43 at mRNA and protein levels was markedly increased with augmented linear distribution in the myocardial cell intercalated disc in TXL group(P<0.05). CONCLUSION: TXL reduces the incidence of VA after MI via inhibiting the Cx43 remodeling.     

Pathophysiologic changes of local ischemic coronary artery and cardiac muscle after ligating canine LAD

AIM: To evaluate the reliability of making a research model of coronary artery stenosis and local myocardial infarction reproduced in dog by ligating canine LAD. METHODS: We disparted 30 aged healthy cross-breed dogs ［(18.5±6.7) kg］ into three groups. The near part of the LAD through left minimal thoracic incision was ligated to interdict 25% (group A), 50% (group B), 75% (group C) of the flux, respectively. The changes of plasma endothelium-derived factors NO, ET-1, sP-selectin and CTnT were measured before ligation and at different time points after ligation. The expression of P-selectin gene in cardiac muscle was detected by Western blotting. The segments of distal parts of the ligated LAD were cut and pathological changes of the patches of topical cardiac muscle were observed by electronic microscope. RESULTS: After ligation, NO/ET-1, P-selectin and CTnT had significant changes in group B (P<0.05) and group C (P<0.01). The expression of P-selectin of cardiac muscle was highly up-regulated after ligating in B (50%) and C (75%) group, In C group animals, a typical far more intense expression pattern was found. Under electronic microscope, the endothelium and other structures of the LAD wall and ultrastructure of myocardial cells had obvious changes in later two groups, especially in group C. There were a typical stenosis of LAD and myocardial infarction. CONCLUSIONS: Ligating the LAD 75% severely damages the endothelial cell and cardiac muscle cells of local ischemic vessel and cardiac muscle, thus forms the typical local stenosis of coronary artery and myocardial infarction, such method is a safe and reasonable way for making a disease model for studying CABG in surgery.     

Relationship between endothelial dysfunction and renin-angiotensin-aldosterone system under the condition of fatigue stress and effect of herbs to dredge collaterals

AIM: To investigate the mechanisms and relationship among endothelial dysfunction, renin-angiotensin-aldosterone system (RAAS) and family of interleukins under the condition of excessive fatigue, by using the rats with fatigue stress. METHODS: Healthy male Wistar rats were randomly divided into control group, homocysteine (HCY) group, fatigue stress group, renshen group, shuangshen group and tongxinluo group. Radioimmunoassay was carried out to detect plasma renin activity (PRA), angiotensin II (AngⅡ), aldosterone (ALD), endothelin (ET), thromboxane-2 (TXA2), prostaglandin I2 (PGI2) in plasma and interleukin-1β, 2, 6, 10 (IL -1β, IL -2, IL -6) in sera. ELISA was used to detect NE and IL -10. The content of nitric oxide (NO) in sera was also detected. The bioinformatical analysis was used to determine the relationship between RAAS and endothelial dysfunction. RESULTS: Compared with control group, the levels of ET-1 and TXA2 in fatigue stress group were significantly increased (P<0.01, P<0.05), but the content of PGI2 and NO was significantly decreased (P<0.01, P<0.01). Compared with control group, the renin activity in plasma of animals in fatigue stress group was significantly decreased (P<0.01), the AngⅡ, IL -1β, IL -6 level was significantly increased compared with the control group (P<0.01, P<0.01), and was significantly increased compared with the HCY group (P<0.05, P<0.01, P<0.01). The NE level showed the tendency of decrease in different degree. After the intervention of three kinds of herbs to dredge collaterals, the ET-1, AngⅡ, IL -1β, IL-6 level in plasma was decreased in different degree (P<0.05, P<0.01, P<0.01), and at the same time, the contents of NO and NE level were significantly increased (P<0.05). The ALD level in tongxinluo groups was apparently higher than that in control group and the fatigue stress group (P<0.05). The bioinformatics analysis showed that Ang II and ET, IL-1; PGI2 and ALD; NO and ALD composed of three subsystems and interrelated according to the principle of optimality of complex system, and gradual change regulation was also observed in fatigue stress group. However, in control group, HCY group and tongxinluo group, the same interrelation among subsystems was not existed. CONCLUSION: In a state of long-term excessive fatigue, vascular endothelial dysfunction may be induced, and is related with renin-angiotensin-aldosterone system imbalance and serious turbulence of the autonomic dysfunction. Herbs to dredge collaterals could improve it significantly. The results suggest that bearing excessive fatigue and pressure in long-term may be the potential risk factors to induce vascular endothelial dysfunction and further result in cardio-cerebro vascular diseases, Tongluo therapy may be one of the useful ways to prevent such diseases.     

Chronic inhibition of cilazapril on pulmonary vascular and myocardial cell proliferation in hypoxic rats

AIM: To explore the mechanism of cilazapril inhibiting proliferation of pulmonary vascular and myocardial cells in hypoxic rats. METHODS: 30 male Wistar rats were used and divided into three groups: normal control (group A)， intermittent hypoxia for 4 weeks (group B) and intermittent hypoxia for 4 weeks plus cilazapril treatment (group C). The cell proliferation and structural remodeling in pulmonary vasculature and myocardium during hypoxia were studied by biochemical analysis, radioimmunoassay, immunohistochemistry, terminal deoxyuridine tripnosphate nick end labeling and correlated with hemodynamic. RESULTS: (1) The mean pulmonary artery pressure (mPAP) and the right ventricle to left ventricle plus ventricular septum ratio (R/L±S) were significantly higher in the hypoxic rat than that in control animals, while increased thickness of the pulmonary vascular wall and vascular lumen with decrease in the caliber as well as myocardial hypertrophy were observed in hypoxic rats. (2) The proliferative index (PI) of pulmonary arteria and myocardium was significantly higher in group B and C than that in group A. The distribution of ET-1 positive cells was seen in pulmonary arterial wall and cardiomyocytes. The ET-1 immunoreactivity was group B>group C>group A by turns. (3) The concentrations of plasma endothelin-1 (ET-1) and angiotensin converting enzyme (ACE) were significantly higher in group B than that in group A. However, the ET-1 and ACE were significantly lower in group C than those in group B. (4) The ET-1 and ACE had a significant positive correlation with R/L+S, mPAP and PI, respectively. The multivariate linear regression analysis revealed that ET-1 and ACE were major factor affecting PI. CONCLUSION: The pulmonary vascular and myocardial structural remodeling are one of the pathogenesis accompanied with excessive cell proliferation in hypoxic pulmonary hypertension (PH). Cilazapril effectively prevents and treats the hypoxic PH by inhibiting cell proliferation and structural remodeling of pulmonary circulation, as induced by ET-1 and ACE.