OBJECTIVE: To investigate the vascular damage effects and possible mechanism of acute exposure to ozone (O) in male Wistar rats. METHODS: One hundred and twenty male Wistar rats were randomly divided into six groups, 20 in each group. The experimental animals were placed in a gas poisoning cabinet, the control group was exposed to filtered air, and the treatment group was exposed to ozone at concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, 2.0 ppm, and 4.0 ppm, respectively, for 4 hours. Arterial blood pressure data were obtained by PC-lab medical physiological signal acquisition system. Blood rheology indicators and blood biochemical indicators were detected by Tianjin Dean Diagnostic Laboratory. Serum endothelin-1 (ET-1), homocysteine (HCY), von Willebrand factor (vWF), 8-hydroxydeoxyguanosine (8-OhdG), interleukin (IL-6) and tumor necrosis factor alpha (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA) microplate assay. Oxidative stress indicators superoxide dismutase (SOD) activity and malondialdehyde (MDA) were determined by xanthine oxidase method, thiobarbituric acid (TBA) method, reduced glutathione (GSH) and nitric oxide (NO) were tested by using microplate colorimetry. Paraffin sections were prepared from thoracic aorta tissue, and vascular structure was observed by HE staining. RESULTS: Acute exposure to 0.12 ppm ozone could cause a significant increase in arterial systolic blood pressure (SBP). Exposure to different concentrations of ozone could cause a significant increase in plasma viscosity, and the K value of the ESR equation was significantly increased in the 1.0 ppm ozone exposure group. Both the relative and reduced viscosities were significantly reduced at ozone concentrations of 0.5 ppm and 4.0 ppm, while the red blood cell deformation index was increased significantly at ozone concentrations of 0.12 ppm, 0.5 ppm, 1.0 ppm, and 2.0 ppm. Acute ozone exposure resulted in the decrease of total cholesterol content. The content of high-density lipoprotein cholesterol (HDL-C) was significantly reduced in the 0.12 ppm ozone exposure group. When the ozone concentration was higher than 1.0 ppm, the body may also had an inflammatory reaction (increased TNF-α) and oxidative stress (increased MDA, decreased GSH). Acute exposure to ozone could lead to elevated levels of ET-1 in the blood, with significant differences in the 4.0 ppm concentration group, while HCY levels were decreased firstly and then increased, reaching the highest in the 1.0 ppm concentration group. No obvious pathological changes were observed in the thoracic aorta. CONCLUSION Acute ozone exposure can affect arterial blood pressure, blood rheology and cholesterol metabolism in rats. The possible mechanism is that ozone exposure leads to inflammatory reaction and oxidative stress reaction, causing vascular endothelial function damage, and vascular endothelial cells increase with ozone exposure concentration.
Animals ; Blood Vessels ; injuries ; Deoxyguanosine ; analogs & derivatives ; blood ; Endothelin-1 ; blood ; Homocysteine ; blood ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Ozone ; toxicity ; Rats ; Rats, Wistar ; Superoxide Dismutase ; analysis ; Tumor Necrosis Factor-alpha ; blood ; von Willebrand Factor ; analysis

OBJECTIVETo evaluate the efficacy and safety of Tongxinluo Capsule (, TXL) for patients with cardiac syndrome X (CSX).

METHODSRandomized controlled trials (RCTs) regarding TXL in the treatment of CSX were searched in Chinese Biomedicine Literature Database, China National Knowledge Infrastructure, Chinese Scientific Journal Database, Wanfang Database, PubMed, EMBASE, Cochrane Central Register of Controlled Trial, websites of the Chinese and International Clinical Trial Registry platform up to June 30, 2015. The intervention was either TXL alone or TXL combined with conventional treatment, while the control intervention was conventional treatment with or without placebo. Data extraction, methodological quality assessment and data analyses were performed according to the Cochrane criteria. The primary outcome was a composite event of death, acute myocardial infarction (AMI), angina requiring hospitalization, revascularization, and heart failure. The secondary outcome measures were angina symptom improvement, electrocardiograph (ECG) improvement, and serum endothelin-1 (ET-1) level. The adverse events were also recorded. RevMan 5.3 software was applied for data analyses.

RESULTSTwelve RCTs (696 patients) were included. Compared with conventional treatment, the addition of TXL to conventional treatment showed some benefits on relieving angina symptoms [risk ratio (RR): 1.46, 95% confidence interval (CI) (1.25, 1.71), P<0.01], and improving ECG [RR: 1.45, 95% CI (1.21, 1.74), P<0.01]. The pooled result did not support a benefit of TXL on reducing the incidence of primary outcome [RR: 0.20, 95% CI (0.02, 1.61), P=0.13]. In addition, TXL decreased serum ET-1 concentration of CSX patients [standardized mean number:-1.63, 95% CI (-2.29,-0.96), P<0.01]. No serious adverse events were reported.

CONCLUSIONSTXL documents potential benefits on attenuating angina symptoms, improving ECG and decreasing serum ET-1 level for CSX patients. However, more rigorous RCTs with high quality are needed to confirm its efficacy and safety.

Capsules ; Cardiovascular Diseases ; diagnostic imaging ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Electrocardiography ; Endothelin-1 ; blood ; Humans ; Middle Aged ; Outcome Assessment (Health Care) ; Publication Bias ; Syndrome

This study aimed to investigate the antihypertensive effect and possible mechanism of Dendrobium officinale flos on hypertensive rats induced by high glucose and high fat compound alcohol. The hypertensive models were successfully made by high-glucose and high-fat diet, with gradient drinking for 4 weeks, and then divided into model control group, valsartan (5.7 mg·kg⁻¹) positive control group and D. officinale flos groups (3，1 g·kg⁻¹). After 6 weeks of treatment, the blood pressure of rats was measured regularly. After the last administration, endothelin-1 (ET-1), thromboxane B₂ (TXB₂), prostacyclin (PGI₂) and nitric oxide (NO) were tested. Endothelial nitric oxide synthase (eNOS) expression and lesion status in thoracic aorta were detected. The vascular endothelium dependent dilation of the thoracic aorta was detected by the isolated vascular loop tension test. The results showed that D. officinale flos could significantly reduce systolic blood pressure and mean arterial pressure in hypertensive rats, inhibit the thickening of thoracic aorta and the loss of endothelial cells, reduce plasma content of ET-1 and TXB₂, and increase the content of PGI₂ and NO. After long-term administration, vascular endothelium dependent dilation of the thoracic aorta was significantly increased, and could be blocked by the eNOS inhibitor (L-NAME) and increase the expression of eNOS. Therefore, D. officinale flos has an obvious antihypertensive effect on high glucose and high fat compound alcohol-induced hypertensive rats. Its mechanism may be correlated with the improvement of vascular diastolic function by protecting vascular endothelial cells, and finally resist hypertension.
Animals ; Antihypertensive Agents ; pharmacology ; Blood Pressure ; Dendrobium ; chemistry ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Endothelin-1 ; blood ; Endothelium, Vascular ; drug effects ; Epoprostenol ; blood ; Glucose ; Hypertension ; chemically induced ; drug therapy ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; T-Box Domain Proteins ; blood ; Vasodilation

OBJECTIVETo observe the effect of electroacupuncture (EA) stimulation on the expressions of angiotensinogen (AGT), angiotensin II type 1 receptor (AT1R), endothelin-1 (ET1), and endothelin A receptor (ETAR) mRNA in spontaneously hypertensive rat (SHR) aorta.

METHODSEighteen male SHRs were randomly divided into three groups, an SHR group, an SHR Baihui (DU 20) and Zusanli (ST 36) acupoint (SHR-AP) group, and an SHR non-acupoint (SHR-NAP) group, with 6 rats in each group. Six Wistar rats were used as a control. Rats in the SHR-AP group were stimulated by DU 20 and ST 36 acupoints, both of which were connected with EA. EA was handled one time every Monday, Wednesday and Friday, for total 24 times (8 weeks). SHRNAP rats were acupointed at a 15°angle flat into 0.5 cm to two points, which were 1 and 2 cm from rail tip separately. EA parameters were the same as the SHR-AP rats. SHR control rats and Wistar rats were fixed without EA. Real-time quantitative polymerase chain reaction (PCR) was used to measure AGT, AT1R, ET1, and ETAR mRNA expression in rat aorta.

RESULTSEA stimulation significantly reduced rat aorta vascular AGT, ET1, ETAR and AT1R mRNA expressions in the SHR-AP and SHR-NAP groups (P <0.01). Among these four genes, AT1R mRNA expression was significantly lower in the SHR-AP than in the SHR-NAP group (P <0.01).

CONCLUSIONEA could reduce the AT1R mRNA expression in SHR-AP rat aorta, indicating a potential mechanism for the hypotensive effects of EA.

Angiotensinogen ; genetics ; metabolism ; Animals ; Aorta ; metabolism ; physiopathology ; Blood Pressure ; Electroacupuncture ; Endothelin-1 ; genetics ; metabolism ; Gene Expression Regulation ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats, Inbred SHR ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Receptor, Endothelin A ; genetics ; metabolism

OBJECTIVETo observe the effect of Tripterygium glycosides (TG) on the expression of hypoxia-inducible factor-1α and endothelin-1 in the kidney of diabetic rats and explore the possible mechanism underlying the protective effect of TG against diabetic nephropathy.

METHODSSixty 8-week-old male SD rats were randomly divided into normal control group (n=10) and streptozotocin-induced diabetes mellitus (DM) model group (n=50). The diabetic model rats were then randomly divided into DM group, low-dose (8 mg/kg) and high-dose (16 mg/kg) TG treatment groups, and Irbesartan (50 mg/kg) treatment group. After 8 weeks, the levels of blood glucose (BG), 24-h urine protein (24 h Upro), serum creatinine (Scr) and blood urea nitrogen (BUN) were measured. The pathological changes in the renal tissues were examined by optical microscopy, and the mean glomerular area (MGA) and mean glomerular volume (MGV) were measured with pathological image analysis. Immunohistochemical and Western blotting were used to detect the expression of HIF-1α and ET-1 protein in the renal tissue, and their mRNA expressions were detected using real-time fluorescence quantitative PCR.

RESULTSHIF-1α and ET-1 expression increased in the kidney of diabetic rats. Compared with the diabetic model rats, the rats receiving TG and Irbesartan treatment showed decreased levels of Scr, BUN, 24h Upro, MGA and MGV, improved renal histopathology, and reduced expression of HIF-1α and ET-1 mRNA and protein in the renal tissue. These changes were more obvious in high-dose TG treatment group. Correlation analysis showed that the expression of HIF-1α was positively correlated with that of ET-1, and they were both positively correlated with kidney weight index (KW/BW), 24 h Upro, MGA, and MGV.

CONCLUSIONHIF-1α and ET-1 are overexpressed in the kidney of diabetic rats. TG can improve kidney damage in diabetic rats and delay the development of diabetic nephropathy by inhibiting the HIF-1α and ET-1 expression.

Animals ; Biphenyl Compounds ; pharmacology ; Blood Glucose ; Blood Urea Nitrogen ; Creatinine ; blood ; Diabetes Mellitus, Experimental ; metabolism ; Endothelin-1 ; metabolism ; Glycosides ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Kidney ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tetrazoles ; pharmacology ; Tripterygium ; chemistry

OBJECTIVETo examine the effect of H2S donor, sodium hydrosulfide (NaHS), on ET-1 level in plasma and aorta in rats with atherosclerosis (AS).

METHODThirty male rats, weighting 200-220 g, were randomly divided into AS, AS+NaHS and control groups, n = 10 in each group.Rats were given a single dose of vitamin D3 (700 000 U/kg) in the first three days and fed with a high-cholesterol diet for 8 weeks to induce AS. Rats in AS+NaHS group were intraperitoneally injected with an H2S donor NaHS, at a dose of 56 µmol/(kg·d) for 8 weeks. At the end of the experiment for 8 weeks, all the rats were sacrificed. The plasma was collected and the aorta and coronary tissues were isolated. The atherosclerotic lesions in both aorta and coronary arteries were detected using oil red O method. H2S concentration in plasma was determined with sulfide-sensitive electrode method. ET-1 levels in plasma and aorta were calculated by radioimmunoassay kit and the localization of ET-1 in the aorta was detected by immunohistochemistry. Plasma nitric oxide synthase (NOS), endothelial NOS (eNOS), inducible NOS (iNOS) were detected with colorimetry.

RESULTAS plaque area in root of aorta of rats in AS group, AS+NaHS group and control group were (11.6±3.3)%, (1.6±1.1)%, (0.0±0.1)% respectively. The difference in AS plaque area in root of aorta among the three groups was statistically significant (F=97.675, P < 0.05). AS plaque area in coronary artery of rats in AS group, AS+NaHS group and control group were (21.4±5.7)%, (4.8±2.5)%, (0.0±0.0)% respectively. The difference in AS plaque area in coronary artery among the three groups was statistically significant (F=97.519, P < 0.05). Plasma H2S level in rats of AS group ((22.0±3.1) µmol/L) was significantly lower than that of control group ((27.9±1.0) µmol/L) and AS+NaHS group ((33.3±6.2) µmol/L, all P < 0.05). Compared with control group ((70.0±10.7) ng/L), plasma ET-1 in rats of AS group ((89.6±14.2) ng/L) and AS+NaHS group ((93.1±15.5) ng/L, P both < 0.05) were increased. However, there was no significant difference in plasma ET-1 content in rats between AS+NaHS group and AS group (P > 0.05). Compared with control group ((3.8±1.2) ng/g), ET-1 content in aorta in rats of AS group ((11.9±4.9) ng/g) and AS+NaHS group ((8.2±2.5) ng/g, both P < 0.05) were increased, and ET-1 content in aorta in rats of AS+NaHS group was decreased compared with AS group (P < 0.05). Immunochemistry results showed that ET expression in cytoplasm in aortic endothelial cells in rats of AS group was strengthened, while ET expression in rats of control group and AS+NaHS group was weak. NOS activity of rats in control group, AS group and AS+NaHS group was (25.4±5.6), (51.8±10.0) and (27.6±6.5) U/ml, eNOS activity (15.3±6.2), (4.5±2.7) and (8.7±3.9) U/ml, and iNOS activity (9.9±4.0), (47.3±10.7) and (19.0±5.2) U/ml, respectively.Differences among the three groups were statistically significant (NOS activity: F=37.231, P < 0.05, eNOS activity: F=14.600, P < 0.05, and iNOS activity: F=72.131, P < 0.05).

CONCLUSIONH2S donor NaHS reduced the AS plaque in AS rats. The mechanisms might involve the protective effect of H2S on the vascular endothelial cell, decreasing ET-1 production in aortal endothelium of atherosclerotic rats.

Animals ; Aorta ; metabolism ; pathology ; Atherosclerosis ; metabolism ; pathology ; Coronary Vessels ; pathology ; Disease Models, Animal ; Endothelin-1 ; blood ; metabolism ; Hydrogen Sulfide ; pharmacology ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Random Allocation ; Rats ; Sulfides ; pharmacology

Recent studies showed that pathology of alcoholic encephalopathy was associated with cerebral vascular damage. TMP (tetramethyl- pyrazine) is widely used in the treatment of cerebrovascular diseases, however, it has not been reported whether TMP can relieve alcohol-induced cerebral vascular damages. The study was performed to investigate the learning and memory, cerebrovascular pathological changes and the expressions of vascular endothelial growth factor (VEGF) and serum levelsofendothelin-1 (ET-1) in the rat model of chronic alcoholic encephalopathy, and explore the effects of TMP intervention on alcoholic encephalopathy. In the present study, the rat model of chronic alcoholic encephalopathy was established by the gavage administration of alcohol; the learning and memory ability was tested by Morris water maze; the expression of VEGF was measured by RT-PCR and Western blot; and the serum levels of ET-1 was measured by radioimmunoassay. We found that alcohol intoxication impaired learning and memory, induced VEGF overexpression and increased ET 1 concentrations. TMP intervention improved learning abilities, increased the VEGF expression and reduced ET-1 level. These results indicate that TMP exhibits therapeutic effects on chronic alcoholic encephalopathy.
Alcohol-Induced Disorders, Nervous System ; complications ; drug therapy ; physiopathology ; Animals ; Cerebrovascular Circulation ; drug effects ; Disease Models, Animal ; Endothelin-1 ; blood ; Learning ; drug effects ; Male ; Memory ; drug effects ; Pyrazines ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; analysis ; Vasodilator Agents ; pharmacology ; therapeutic use

Pulmonary arterial hypertension (PAH) causes right ventricular failure due to a gradual increase in pulmonary vascular resistance. The purposes of this study were to confirm the engraftment of human umbilical cord blood-mesenchymal stem cells (hUCB-MSCs) placed in the correct place in the lung and research on changes of hemodynamics, pulmonary pathology, immunomodulation and several gene expressions in monocrotaline (MCT)-induced PAH rat models after hUCB-MSCs transfusion. The rats were grouped as follows: the control (C) group; the M group (MCT 60 mg/kg); the U group (hUCB-MSCs transfusion). They received transfusions via the external jugular vein a week after MCT injection. The mean right ventricular pressure (RVP) was significantly reduced in the U group after the 2 week. The indicators of RV hypertrophy were significantly reduced in the U group at week 4. Reduced medial wall thickness in the pulmonary arteriole was noted in the U group at week 4. Reduced number of intra-acinar muscular pulmonary arteries was observed in the U group after 2 week. Protein expressions such as endothelin (ET)-1, endothelin receptor A (ERA), endothelial nitric oxide synthase (eNOS) and matrix metalloproteinase (MMP)-2 significantly decreased at week 4. The decreased levels of ERA, eNOS and MMP-2 immunoreactivity were noted by immnohistochemical staining. After hUCB-MSCs were administered, there were the improvement of RVH and mean RVP. Reductions in several protein expressions and immunomodulation were also detected. It is suggested that hUCB-MSCs may be a promising therapeutic option for PAH.
Animals ; Cytokines/metabolism ; Disease Models, Animal ; Endothelin-1/metabolism ; Fetal Blood/*cytology ; Gene Expression Regulation/drug effects ; Hemodynamics ; Humans ; Hypertension, Pulmonary/chemically induced/*therapy ; Hypertrophy, Right Ventricular/physiopathology ; Immunohistochemistry ; Lung/metabolism/pathology ; Male ; Matrix Metalloproteinase 2/metabolism ; *Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells/*cytology/metabolism ; Monocrotaline/toxicity ; Nitric Oxide Synthase Type III/metabolism ; Pulmonary Artery/pathology ; Rats ; Rats, Sprague-Dawley ; Receptor, Endothelin A/metabolism

OBJECTIVETo investigate the effects of different anesthesia depth on stress response in elderly patients undergoing elective laparoscopic surgery for colorectal cancer.

METHODSA total of 105 ASA I-III patients aged 60-91 years undergoing elective laparoscopic surgery for colorectal cancer with general anesthesia were randomized into 3 groups, namely group A with a target Narcotrend index (NI) maintained at D0 level, group B with a NI at D2 level, and group C with a NI at E1 level. The anesthetics (profopol and remifentanil) were adjusted according to Narcotrend monitoring results to maintain the specified anesthesia depth. The patients' heart rate (HR) and mean artery pressure (MAP) were recorded before anesthesia (T0), before intubation (T1), immediately after intubation (T2), at 2 min before pneumoperitoneum (T3), 2 min after pneumoperitoneum (T4), at the end of the surgery (T5) and extubation (T6). Serum levels of cortisol, adrenocorticotropic hormone (ACTH), endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and C-reactive protein (CRP) were measured by standard ELISA and radioimmunoassay before anesthesia (Ta), at the end of the surgery (Tb) and 1 day after the surgery (Tc).

RESULTSHR and MAP in group A increased significantly at T2, T4, and T6 compared to those at T0 (P<0.05), and were higher than those in group B and group C (P<0.05). The MAP in all the 3 groups all decreased at T1 and T3 (P<0.05 or P<0.01), and was markedly lower in group C than in groups A and B (P<0.05). The incidence of hypertension was significantly higher in group A than in groups B and C (P<0.05), while the incidence of hypotension was much higher in group C (P<0.01). There were no obvious differences in serum levels of cortisol, ACTH, CRP, IL-6, TNF-a, or ET-1 among the groups at Ta (P>0.05). The serum levels of ACTH in the 3 groups all significantly increased at Tb and Tc (P<0.01). CRP, IL-6 and TNF-a levels in group A were increased at Tb and Tc (P<0.05 or P<0.01) and significantly higher than those in groups B and C (P<0.05 or P<0.01). Cortisol in groups A and B increased at Tb and Tc (P<0.05) to a significantly higher level than that in group C (P<0.01). ET-1 level in group C at Tb and Tc was lower than those in groups A and B (P<0.05 or P<0.01).

CONCLUSIONSMaintaining the anesthesia depth for a NI at the D2 and E1 level can both attenuate the stress response in elderly patients undergoing laparoscopic surgery for colorectal cancer, but the hemodynamic stability can be better at a D2 level.

Adrenocorticotropic Hormone ; blood ; Aged ; Aged, 80 and over ; Anesthesia, General ; methods ; Blood Pressure ; C-Reactive Protein ; metabolism ; Colorectal Neoplasms ; surgery ; Elective Surgical Procedures ; Endothelin-1 ; blood ; Heart Rate ; Humans ; Hydrocortisone ; blood ; Interleukin-6 ; blood ; Laparoscopy ; Middle Aged ; Piperidines ; administration & dosage ; Propofol ; administration & dosage ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the relationship between non-alcoholic steatohepatitis (NASH) and atherosclerosis in young and middle-aged patients, and to provide clinical evidence that will aid in prevention and prediction of development of atherosclerosis or cardiovascular diseases in patients with non-alcoholic fatty liver disease (NAFLD).

METHODSFifty-one patients with biopsy-proven NAFLD (18 to 60 years in age) were divided into two groups: cases with simple non-alcoholic fatty liver (NAFL, n = 11) and cases with NASH (n = 40). All subjects underwent physical examination and anthropometric measurements. Fasting serum was assayed by blood biochemistry. Insulin resistance was estimated by the homeostatic model assessment index (HOMA-IR). Serum levels of high-sensitivity C-reactive protein (hs-CRP), soluble intercellular adhesion molecule-1 (sICAM-1), and endothelin-1 (ET-1) were detected by enzyme-linked immunosorbent assay. Carotid intima-media thickness (CIMT) was estimated by carotid ultrasound. Brachial-ankle pulse wave velocity (baPWV) and ankle-brachial index were estimated using a volume-plethysmographic apparatus. Data for the two groups were summarized as mean +/- SD or interquartile range and intergroup differences were evaluated by paired t-test or Wilcoxon test, with two-sided P-values less than 0.05 indicating significance.

RESULTSThe serum levels of hs-CRP, sICAM-1, and ET-1 were significantly higher in the NASH group than the NAFL group (all P less than 0.001). In addition, the CIMT and baPWV were significantly higher in the NASH group than the NAFL group (both P less than 0.05). The HOMA-IR was also significantly higher in the NASH group than the NAFL group (P less than 0.001).

CONCLUSIONLiver inflammation and insulin resistance may play important, and possibly collaborative, roles in promoting arterial endothelial dysfunction and atherosclerosis in NAFLD patients. NASH patients, especially those who are young and middle-aged, may benefit from early monitoring and prevention strategies to help decrease the risk of developing severe cardiovascular diseases.

Adolescent ; Adult ; Atherosclerosis ; pathology ; C-Reactive Protein ; metabolism ; Carotid Arteries ; diagnostic imaging ; Carotid Intima-Media Thickness ; Endothelin-1 ; blood ; Female ; Humans ; Insulin ; blood ; Insulin Resistance ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; blood ; diagnostic imaging ; Young Adult