[Efficacy and safety of percutaneous closure of ventricular septal rupture after acute myocardial infarction: a clinical study of 69 cases].

Objective: To investigate the efficacy and safety of percutaneous closure of ventricular septal rupture (VSR) after acute myocardial infarction (AMI) and the risk factors of all-cause mortality at 30 days after operation. Methods: This is a retrospective case series study. A total of 69 patients with post-AMI VSR, underwent percutaneous closure of VSR from October 2013 to May 2020 in Department of Cardiology of Henan Provincial People's Hospital and Department of Cardiology of Central China Fuwai Hospital, were included. Patients were divided into survival group (53 cases) and non-survival group (16 cases) according to the status at 30 days after operation. Clinical data were collected and analyzed during hospitalization. Telephone follow-up was performed 30 days after operation. The primary safety endpoint was occlusion failure and all-cause mortality at 30 days post operation. The secondary safety endpoint was the operation related or non-operation related complications. Efficacy endpoint included NYHA classification of cardiac function, index measured by right heart catheterization and echocardiography. Multivariate logistic regression was performed to analyze the risk factors of all-cause mortality at 30 days after operation. Results: A total of 69 patients, aged 67 (64, 71) years, including 42 women (60.9%), were enrolled in this study. All-cause death occurred in 16 patients (23.2%), including 13 in-hospital death and 3 death during follow-up. There were 4 cases of closure failure (5.8%). Among the 65 patients with successful closure, 12 (18.5%) experienced operation-related complications, among which 8 (12.3%) experienced valve injury. The mortality was significantly higher in patients with operation-related complications than that in patients without operation-related complications (41.7% (5/12) vs. 13.2% (7/53), P = 0.022). One case received percutaneous closure of VSR and PCI, this patient experienced new-onset AMI immediately post procedure and died thereafter (1.5%). One case (1.5%) developed multiple organ failure and 2 cases (3.1%) developed gastrointestinal bleeding post operation. All of the 65 patients with successful occlusion completed postoperative echocardiography, 56 patients completed cardiac function assessment at discharge, and 53 patients who survived up to 30 days post discharge completed clinical follow up by telephone. The NYHA cardiac function at discharge and 30 days after operation were significantly improved as compared to that before operation (P<0.001), the ratio of NYHA Ⅰ and Ⅱ patients was significantly higher post operation at these two time points as compared to baseline level (76.8% (43/56) vs. 23.1% (15/65), P<0.001, 77.4% (41/53) vs. 23.1% (15/65), P<0.001). The pulmonary circulation/systemic circulation blood flow ratio (Qp/Qs), pulmonary artery systolic pressure (PASP) and left ventricular end-diastolic diameter (LVDd) were decreased, aortic systolic pressure (ASP) and left ventricular ejection fraction (LVEF) were increased post operation (P<0.05). Multivariate logistic regression analysis showed that WBC>9.8×109/L (OR=20.94, 95%CI 1.21-362.93, P=0.037) and NT-ProBNP>6 000 ng/L (OR=869.11, 95%CI 2.93-258 058.34, P=0.020) were the independent risk factors of mortality at 30 days. Conclusions: Percutaneous closure in VSR after AMI is safe and effective. The increase of WBC and NT-ProBNP are the independent risk factors of all-cause mortality at 30 days after operation.

Analyses of the Tympanic Membrane Impulse Response Measured with High-Speed Holography.

The Tympanic Membrane (TM) transforms acoustic energy to ossicular vibration. The shape and the displacement of the TM play an important role in this process. We developed a High-speed Digital Holography (HDH) system to measure the shape and transient displacements of the TM induced by acoustic clicks. The displacements were further normalized by the measured shape to derive surface normal displacements at over 100,000 points on the TM surface. Frequency and impulse response analyses were performed at each TM point, which enable us to describe 2D surface maps of four new TM mechanical parameters. From frequency domain analyses, we describe the (i) dominant frequencies of the displacement per sound pressure based on Frequency Response Function (FRF) at each surface point. From time domain analyses, we describe the (ii) rising time, (iii) exponential decay time, and the (iv) root-mean-square (rms) displacement of the TM based on Impulse Response Function (IRF) at each surface point. The resultant 2D maps show that a majority of the TM surface has a dominant frequency of around 1.5 kHz. The rising times suggest that much of the TM surface is set into motion within 50 µs of an impulsive stimulus. The maps of the exponential decay time of the IRF illustrate spatial variations in damping, the least known TM mechanical property. The damping ratios at locations with varied dominant frequencies are quantified and compared.

Controlling nutritional status (CONUT) score-based nomogram to predict overall survival of patients with HBV-associated hepatocellular carcinoma after curative hepatectomy.

PURPOSE: As a novel immune-nutritional biomarker, the controlling nutritional status (CONUT) score has been reported to predict outcomes in cancer patients. We aimed to elucidate the prognostic value of preoperative CONUT score and construct a CONUT score-based nomogram to predict individual survival of patients with hepatitis B viral (HBV)-associated hepatocellular carcinoma (HCC) after curative hepatectomy. METHODS: Preoperative CONUT score was retrospectively calculated in 380 HBV-associated HCC patients undergoing radical resection between 2006 and 2012. Patients were assigned to two groups: CONUT-low ( < 2) and CONUT-high ( ≥ 2), according to the optimal cut-off value determined using receiver operating characteristic analysis. Associations of CONUT score with oncological outcomes were evaluated. The Cox proportional hazard model was used to identify predictors of survival and a new nomogram was developed based on the independent prognostic factors for overall survival (OS). RESULTS: The CONUT score exhibited a higher area under the curve value than the other immune-nutritional parameters. The CONUT-high group had significant poorer OS and recurrence-free survival compared with CONUT-low group (P < 0.001 and P = 0.016, respectively). Multivariate analyses identified CONUT score, liver cirrhosis, tumor size and differentiation as independent prognostic factors for OS. And the nomogram based on these four variables had superior discriminative ability to predict survival compared with other conventional staging systems. CONCLUSIONS: Preoperative CONUT score is an effective independent predictor of OS in patients with resected HBV-related HCC. This novel nomogram based on CONUT may provide accurate and individualized survival prediction for HCC patients undergoing surgical resection.

[Association between Lipoprotein(a) and the characteristics of left main coronary artery plaque in patients with stable angina pectoris].

Objective: To investigate the relationship between Lipoprotein (LP) (a) level and the characteristics of tissue components of left main coronary artery (LMCA) plaque. Methods: A total of 102 patients with stable angina pectoris who underwent percutaneous coronary intervention (PCI) in the People's Hospital of Henan Province from June 2010 to October 2016 were included. We performed intravascular ultrasound-virtual histology (IVUS-VH) to their LMCAs and evaluated the tissue characteristics, and the blood level of total cholesterol (TC), triacylglycerol (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), ApoB, ApoA1, LP(a) were measured. According to the value of their LP(a) level they were divided into 2 groups (high LP(a) group (>300 mg/L) (n=35) and low LP(a) group (≤300 mg/L) (n=67)), then the relationship between the above lipid values and the tissue characteristics of the LMCA plaque in the patients were evaluated. Results: Patients with a high LP(a) had a larger percentage of fibrolipid volume and a smaller percentage fibrous volume compared to patients with a normal LP(a) (25%±5% vs 13%±6%, P<0.01 and 50%±8% vs 61%±9%, P<0.01). Using multivariate linear regression analysis after adjustment for the above-mentioned confounding factors, LP(a) had a significantly positive correlation with fibrolipid volume percentage (r=0.645, ß=0.29, P<0.01), and had a negative correlation with fibrous volume percentage (r=-0.467, ß=-0.32,P<0.01), suggesting that the LP(a) was associated with the vulnerability of the LMCA plaque. Conclusion: For the patients with stable angina pectoris, the LP(a) has a significantly positive correlation with the percentage of fibrolipid volume and a negative correlation with the percentage of fibrous volume, suggesting that the LP(a) could predict the vulnerability of the LMCA plaque.

[Effect of miR-19b on the proliferation and apoptosis of P19CL6 cells during the late-stage of cardiac differentiation].

Objective: To explore the effect of miR-19b on the function of P19CL6 cells and its molecular mechanism. Methods: Overexpression of miR-19b was carried out by transfecting miR-19b plasmid into the P19CL6 cells. MTT assay and flow cytometry were used to determine cell growth and apoptosis, respectively. Western blot was used to detect the expression level of Sox6 in P19CL6 cells. ELISA assay was used to detect the expression levels of apoptosis-related genes (Bax, Bcl-2) in P19CL6 cells at late-stage cardiac differentiation. Further online software TargetScan was used to predict the target genes of miR-19b and verified by dual luciferase reporter assay. Results: Our data showed that overexpression of miR-19b in P19CL6 cells significantly increased the cell growth rates and the apoptosis inhibition rates. The ratio of apoptosis-related proteins (Bax/Bcl-2) was significantly reduced. Results from the TargetScan and dual luciferase reporter showed that Sox6 is the direct target of miR-19b. Conclusions: We conclude that miR-19b might promote cell proliferation and inhibits cell apoptosis during the late-stage of cardiac differentiation by targeting Sox6 expression.

[Association between the ApoB/A1 ratio and the vulnerability of LMCA plaque in the patients with stable angina pectoris].

Objective: To investigate the relationship between ApoB/A1 ratio and the characteristics of tissue components of their left main coronary artery(LMCA)plaque. Methods: A total of 98 patients with stable angina pectoris who received chronic statin treatment underwentpercutaneous coronary intervention in the People's Hospital of Henan Province from June 2010 to June 2016 were included.We prospectively performed intravascular ultrasound virtualhistology (IVUS-VH) to their LMCA and evaluated the tissue characteristics, and the blood level of total cholesterol (TC) and low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), triglyceride(TG), LDL-C/HDL-C ratio, ApoB, ApoA1, ApoB/A1 ratio were measured, then the association of the tissue characteristics with the aboved lipids values were analyzed. Results: According to the median value of their ApoB/A1ratios (0.80), they were divided into 2 groups [high ApoB/A1 ratio (>0.80) (n=49) and low ApoB/A1 ratio (≤0.80) (n=49)]. The patients with a high ApoB/A1 ratio had alarger fibrolipid volume and a smaller fibrous volume compared to patients with a low ApoB/A1 ratio ( 17.5%±1.2% vs 9.0%±1.0%, P=0.03 and 55.1%±2.1% vs 63.9%±1.8%, P<0.01). Using multivariate linear regression analysis after adjustment for the above-mentioned confounding factors, the ApoB/A1 ratio had a significantly positive correlation with fibrolipid volume (r=0.445, ß=0.29, P=0.010)and had a negative correlation with fibrous volume (r=-0.567, ß=-0.32, P=0.011), suggesting that the ApoB/A1 ratio was associated with the vulnerability of the LMCA plaque. Conclusion: For the patients with stable angina pectoris and chronic treatment of statins, a high ApoB/A1 ratio is associated with a high percentage of fibrolipid volume and a low percentage of fibrous volume in LMCA lesions, suggesting that the ApoB/A1 ratios could predict the vulnerability of the LMCA plaque.

Calcitonin alleviates hyperalgesia in osteoporotic rats by modulating serotonin transporter activity.

Calcitonin may relieve pain by modulating central serotonin activity. Calcitonin partly reversed the hypersensitivity to pain induced by ovariectomy. This suggests that the anti-nociceptive effects of calcitonin in the treatment of osteoporosis may be mediated by alterations in neural serotonin transporter (SERT) activity. INTRODUCTION: This study used a rat model of osteoporosis to evaluate the role of the cerebral serotonin system in the anti-nociceptive effect of calcitonin, a drug used to treat post-menopausal osteoporosis. METHODS: Osteoporosis was induced in rats by ovariectomy (OVX). Rats were then randomized to the following four groups: sham operation, OVX, OVX plus calcitonin, or OVX plus alendronate. RESULTS: OVX led to alterations in bone micro-architecture; alendronate strongly reversed this effect, and calcitonin moderately reversed this effect. OVX increased hyperalgesia (determined as the time for hind paw withdrawal from a heat source); calcitonin reduced this effect, but alendronate had no effect. OVX increased the expression of c-Fos (a neuronal marker of pain) in the thalamus; calcitonin strongly reversed this effect, and alendronate moderately reversed this effect. OVX also reduced SERT but increased 5-HT1A receptor expression and activity; calcitonin aggravated this effect, but alendronate had no effect on recovery of SERT/5-HT1A activity and expression. CONCLUSIONS: Our study of a rat model of osteoporosis suggests that OVX-induced enhancement of the serotonergic system may protect against hyperalgesia. However, the anti-nociceptive effects of calcitonin in osteoporosis may be mediated by decreased neural SERT activity and increased activation of 5-HT1 receptors in the thalamus.

Hepatic carcinoma-associated fibroblasts induce IDO-producing regulatory dendritic cells through IL-6-mediated STAT3 activation.

Although carcinoma-associated fibroblasts (CAFs) in tumor microenvironments have a critical role in immune cell modulation, their effects on the generation of regulatory dendritic cells (DCs) are still unclear. In this study, we initially show that CAFs derived from hepatocellular carcinoma (HCC) tumors facilitate the generation of regulatory DCs, which are characterized by low expression of costimulatory molecules, high suppressive cytokines production and enhanced regulation of immune responses, including T-cell proliferation impairment and promotion of regulatory T-cell (Treg) expansion via indoleamine 2,3-dioxygenase (IDO) upregulation. Our findings also indicate that STAT3 activation in DCs, as mediated by CAF-derived interleukin (IL)-6, is essential to IDO production. Moreover, IDO inhibitor, STAT3 and IL-6 blocking antibodies can reverse this hepatic CAF-DC regulatory function. Therefore, our results provide new insights into the mechanisms by which CAFs induce tumor immune escape as well as a novel cancer immunotherapeutic approach (for example, targeting CAFs, IDO or IL-6).

Optimization of a lensless digital holographic otoscope system for transient measurements of the human tympanic membrane.

In this paper, we propose a multi-pulsed double exposure (MPDE) acquisition method to quantify in full-field-of-view the transient (i.e., >10 kHz) acoustically induced nanometer scale displacements of the human tympanic membrane (TM or eardrum). The method takes advantage of the geometrical linearity and repeatability of the TM displacements to enable high-speed measurements with a conventional camera (i.e., <20 fps). The MPDE is implemented on a previously developed digital holographic system (DHS) to enhance its measurement capabilities, at a minimum cost, while avoiding constraints imposed by the spatial resolutions and dimensions of high-speed (i.e., >50 kfps) cameras. To our knowledge, there is currently no existing system to provide such capabilities for the study of the human TM. The combination of high temporal (i.e., >50 kHz) and spatial (i.e., >500k data points) resolutions enables measurements of the temporal and frequency response of all points across the surface of the TM simultaneously. The repeatability and accuracy of the MPDE method are verified against a Laser Doppler Vibrometer (LDV) on both artificial membranes and ex-vivo human TMs that are acoustically excited with a sharp (i.e., <100 µs duration) click. The measuring capabilities of the DHS, enhanced by the MPDE acquisition method, allow for quantification of spatially dependent motion parameters of the TM, such as modal frequencies, time constants, as well as inferring local material properties.

Cerebral klotho protein as a humoral factor for maintenance of baroreflex.

The klotho protein produced by the choroid plexus is known as a humoral factor in central nervous system. Many hormones affecting the baroreflex sensitivity have been introduced in the brain. However, role of klotho in the baroreflex sensitivity is still unknown. Recently, mutations in the klotho gene have been linked to cardiovascular diseases in both animals and human subjects. Also, silencing of brain klotho has been reported to enhance cold-induced elevation of blood pressure. Thus, we investigated the role of klotho in maintenance of central cardiovascular reflex sensitivity. Male Wistar Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs) were used. Either klotho shRNA or scramble shRNA was also ICV-infused into the brains of WKY rats to investigate the role of klotho in brain. Recombinant klotho or rat IgG was infused into the cerebral paraventricle (ICV) of SHRs for further understanding the role of klotho in hypertension. The baroreflex sensitivity was detected using the challenge with a depressor dose of sodium nitroprusside (SNP, 50 µg/kg) or with a pressor dose of phenylephrine (PE, 8 µg/kg). We found that silencing of klotho expression in the brain decreased the baroreflex sensitivity in WKY rats. Also, modulation of the blood pressure for one week altered the cardiovascular homeostasis and resulted in an increased expression of klotho in medulla oblongata. Moreover, the baroreflex sensitivity was restored in SHRs that received recombinant klotho through ICV brain. Thus, klotho is involved in the maintenance of baroreflex sensitivity in the brain.

Rosiglitazone is effective to improve renal damage in type-1-like diabetic rats.

A marked decrease of klotho expression was observed in the kidney of streptozotocin-induced diabetic rats (STZ rats) showing diabetic nephropathy. It has been documented that klotho is the target gene of PPARγ. However, the effect of PPARγ agonist on klotho expression in kidney of STZ rats remains obscure. Thus, we used rosiglitazone (TZD) as PPARγ agonist to investigate the effect on renal dysfunction in STZ rats. Treatment of TZD reversed the lower levels of PPARγ, klotho, and FGFR1 expressions in kidneys of STZ rats without the correction of hyperglycemia. Also, renal functions and structural defeats were improved by TZD treatment. Taken together, oral administration of TZD may improve STZ-induced diabetic nephropathy due to restoration of the expression of klotho axis through an increase in PPARγ expression without changing blood glucose in rats.

Decrease of FGF receptor (FGFR) and interstitial fibrosis in the kidney of streptozotocin-induced diabetic rats.

Fibrosis is the final disorder of end-stage renal disease. Activation of fibroblast growth factor (FGF) 23-klotho axis could suppress renal fibrosis in mice. Also, a marked decrease of klotho expression was observed in the kidney of streptozotocin-induced diabetic rats (STZ rats). However, relation of FGF in renal fibrosis remained unclear. This study was aimed to screen the effect of hyperglycemia on FGF receptor (FGFR) and fibrosis in kidney of rats with diabetic nephropathy and investigate this potential mechanism in cultured Madin-Darby Canine Kidney (MDCK) epithelial cells. STZ rats were used to treat with insulin or phloridzin at the dose sufficient to correct hyperglycemia for understanding the changes of renal dysfunction. The cultured MDCK cells were also used to treat with high glucose, hydrogen peroxide, or tiron in addition to transfection of siRNA to silence the klotho. Both insulin and phloridzin reversed fibrosis and FGFR expressions in kidney of STZ rats. It was confirmed in high glucose-exposed MDCK cells. However, klotho failed to modify the level of FGFR in MDCK cells. Meanwhile, FGFR was restored by tiron in MDCK cells and in diabetic rats without changing blood glucose. In conclusion, interstitial fibrosis and decreased FGFR expression are observed in the kidney of diabetic rats. This change is reversed by tiron without the correction of blood glucose. Also, klotho has no effect on expression of FGFR. Thus, decrease of oxidative stress is useful for the recovery of FGFR expression and improvement of renal fibrosis in type-1 like diabetic rats.

Decrease of hyperglycemia by syringaldehyde in diabetic rats.

Syringaldehyde is one of the active principles from the stems of Hibiscus taiwanensis (Malvaceae) that has been mentioned to lower hyperglycemia. However, the potential mechanisms for this action of syringaldehyde remain obscure. In the present study, we used streptozotocin to induce diabetic rats (STZ-diabetic rats) as type 1-like diabetic rats and fed fructose-rich chow to rats as type 2-like diabetic rats. Then, we performed the postprandial glucose test and applied the hyperinsulinemic euglycemic clamp to investigate the actions of syringaldehyde. Also, the changes of gene expressions of enzyme relating to glucose homeostasis in muscle and liver were characterized. Syringaldehyde significantly decreased the postprandial plasma glucose in rats, while the plasma insulin was not modified by syringaldehyde. The glucose infusion rate (GIR) in fructose chow-fed rats using hyperinsulinemic euglycemic clamp was markedly improved by syringaldehyde. Additionally, repeated administration of syringaldehyde for 3 days in STZ-diabetic rats resulted in a marked reduction of phosphoenolpyruvate carboxykinase (PEPCK) expression in liver and an increased expression of glucose transporter subtype 4 (GLUT 4) in skeletal muscle. Our results suggest that syringaldehyde may increase glucose utilization to lower hyperglycemia in diabetic rats.

Insulin resistance induced by zymosan as a new animal model in mice.

Insulin resistance (IR) is known as a main problem in diabetic disorders. Some animal models for research in IR have been mentioned. Each model shows merit with some disadvantages. Thus, a new animal model for IR is required. The present study used zymosan, a mixture of cell-wall particles from the yeast named Saccharomyces cerevisiae, to establish a new model of IR in mice. Also, we compared the difference of this model with fructose-rich chow-induced model and found some merits of this model. Moreover, we identified that this model induced by zymosan is reversible and IR can be reversed gradually after termination of treatment. Taken together, we suggest zymosan as a useful agent to induce IR through inflammatory pathway in mice.

Activation of receptors δ (PPARδ) by agonist (GW0742) may enhance lipid metabolism in heart both in vivo and in vitro.

It has been documented that cardiac agents may regulate the lipid metabolism through increased expression of PPARδ in cardiac cells. However, the effect on lipid metabolism by direct activation of PPARδ is still unknown. The present study applied specific PPARδ agonist (GW0742) to investigate this point in the heart of Wistar rats and in the primary cultured cardiomyocytes from neonatal rat. Expressions of PPARδ in the heart and cardiomyocytes after treatment with GW0742 were detected using Western blots. The fatty acid (FA) oxidation and the citric acid (TCA) cycle related genes in cardiomyocytes were also examined. In addition, PPARδ antagonist (GSK0660) and siRNA-PPARδ were employed to characterize the potential mechanisms. After a 7-day treatment with GW0742, expressions of PPARδ in the heart were markedly increased. Increased expressions of FA oxidation and TCA cycle related genes were also observed both in vivo and in vitro. This action of GW0742 was blocked by GSK0660 or by siRNA-PPARδ. The obtained results show that activation of PPARδ by GW0742 is responsible for the increase of FA oxidation and TCA cycle related genes in hearts. Role of PPARδ in the regulation of lipid metabolism in heart is then established.

Improvement of obesity by activation of I1-imidazoline receptors in high fat diet-fed mice.

Imidazoline I1-receptor (I1R) is known to regulate the blood pressure, and rilmenidine, as the agonist, is used to treat hypertension in clinics. However, the role of I1R in obesity is still unclear. In the present study, we investigated the changes of obesity by activation of I1R in high fat diet (HFD)-fed mice. Chronic administration of rilmenidine into HFD-fed mice for 8 weeks significantly reduced body weight, which was reversed by efaroxan at the dose sufficient to block I1R. Also, rilmenidine significantly decreased the energy intake of HFD-fed mice. This reduction of energy intake was abolished by efaroxan at the same dosing for blockade of I1R. However, hypothalamic I1R protein expression in HFD-fed mice was markedly lower than that in normal chow-fed mice. In addition, epididymal white adipose tissue (eWAT) cell size in HFD-fed mice was decreased by rilmenidine via the activation of I1R. Moreover, effect of rilmenidine on appetite disappeared in db/db mice. Taken together, we suggest that rilmenidine can improve obesity in HFD-fed mice through an activation of I1R to ameliorate energy intake and eWAT accumulation.

Characterization of the specificity of imidazoline I-1 receptor antibody for subtype of imidazoline receptors in vitro.

Specific antibodies are essential in the study of receptor protein. Gene matching shows that Nischarin (NISCH) is a mouse homologue of human imidazoline receptor antisera-selective (IRAS) protein, a viable candidate for imidazoline I-1 receptor. However, selectivity of this antibody against imidazoline I-2 or imidazoline I-3 receptors remained obscure. At first, an intracerebroventricular (ICV) injection of anti-NISCH antibody blocked the blood pressure lowering action of rilmenidine (I-1 receptor agonist) in spontaneous hypertensive rat (SHR). However, the same injection of anti-NISCH antibody showed no effect in SHR treated with clonidine (α2 agonist). In order to clarify the selectivity of anti-NISCH antibody for each subtype of imidazoline receptors, this anti-NISCH antibody was subjected to the lysate of organs isolated from Wistar rats including cortex, hippocampus, cerebellum, and brain stem as central nervous tissues, and heart, liver, pancreas, skeletal muscle, kidney, prostate, and bladder as peripheral tissues. The results show that anti-NISCH antibody positively reacted with all tissues including heart, pancreas, skeletal muscle, kidney and bladder by Western blot analysis. Also, the blotting spots for anti-NISCH antibody show a concentration-dependent manner. Moreover, anti-NISCH antibody blocked the action of glucose uptake induced by 2-BFI (I-2 receptor agonist) in L6 cells. Taken together, the obtained data suggest that anti-NISCH antibody can be used not only for imidazoline I-1 receptor but also for I-2 and I-3 subtypes in immunoassays.

Role of TNF-α in renal damage in mice showing hepatic steatosis induced by high fat diet.

The present study was designed to investigate the role of TNF-α in renal damage observed in mice with hepatic steatosis. We induced hepatic steatosis in mice using high fat diet and treated mice with ectanercept at the dose sufficient to block TNF-α receptors or vehicle for 1 month. Plasma TNF-α, total cholesterol (TC), triglyceride (TG), LDL-cholesterol (LDL-C), and HDL-cholesterol (HDL-C) were determined at the end of this treatment. Renal damage was identified by histologic observation and the higher of serum blood urea nitrogen (BUN) and creatinine. Also, changes of PPAR-δ in kidney and renal mesangial cell (RMC) were analyzed using Western blot. Plasma TNF-α was markedly raised in mice showing hepatic steatosis. However, the levels of blood lipids (TC, TG, HDL-C, and LDL-C) and TNF-α were not modified by the treatment of etanercept although the hepatic steatosis has been improved. Etanercept shows renal protection from histological identification and recovery of serum BUN and creatinine levels. Moreover, restoration of PPAR-δ expression by etanercept was observed in mice kidney. Direct effect of TNF-α on PPAR-δ expression was also characterized in RMC cell. We suggest that renal damage in mice with hepatic steatosis is mainly induced by increase of TNF-α through the decrease of renal PPAR-δ. Etanercept could block TNF-α receptors to restore PPAR-δ and improve renal function in mice with hepatic steatosis.

Increase of peroxisome proliferator-activated receptor δ (PPARδ) by digoxin to improve lipid metabolism in the heart of diabetic rats.

Increase of peroxisome proliferator-activated receptor δ (PPARδ) expression by digoxin in the heart of diabetic rats has been documented. The present study investigated the mediation of PPARδ in lipid metabolism improved by digoxin in the heart of diabetic rats and in the hyperglycemia-treated cardiomyocytes using the primary cultured cardiomyocytes from neonatal rat. The lipid deposition within the heart section was assessed in diabetic rats by oil red O staining. The fatty acid oxidation genes in cardiomyocytes were also examined. Inhibitor of calcium ions and siRNA-PPARδ were employed to investigate the potential mechanisms. After a 20-day digoxin treatment, the PPARδ expression was elevated in hearts of diabetic rats while the cardiac lipid deposition was reduced. In neonatal cardiomyocytes, digoxin also caused an increase in expressions of PPARδ and fatty acid oxidation genes. But both actions of digoxin were blocked by BAPTA-AM to chelate calcium ions and by siRNA-PPARδ in cardiomyocytes. The obtained results show that increase of PPARδ by digoxin is related to regulation of fatty acid oxidation genes in cardiac cells mediated by calcium-triggered signals.

Mediation of AMP kinase in the increase of glucose uptake in L6 cells induced by activation of imidazoline I-2 receptors.

Recent work using radioactive tracer indicates that activation of imidazoline I2 receptor (I2R) by guanidinium derivatives may increase the glucose uptake in the skeletal muscle. However, the effect of I2R activation on nonradioactive glucose uptake is still unknown. The ability of glucose uptake in cultured L6 cells is then determined using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose (2-NBDG) as a fluorescence indicator. The changes in 5'-AMP-activated protein kinase (AMPK) expression were also identified by Western blot analysis. In the present study, 2-(2-benzofuranyl)-2-imidazoline (2-BFI) is used to stimulate I2R while 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside (AICAR) is applied to activate AMPK directly. Both compounds can increase 2-NBDG in L6 cells in a concentration-dependent manner. Meanwhile, compound C at concentrations sufficient to inhibit AMPK blocked this increase of glucose uptake by 2-BFI or AICAR. However, only 2-BFI-induced glucose uptake action was dose-dependently blocked by BU224, a specific I2R antagonist, in L6 cells. Moreover, AMPK phosphorylation was markedly increased by 2-BFI or AICAR in L6 cells. Similarly, only the effect of 2-BFI was attenuated by BU224 in L6 cells. Thus, we suggest that AMPK is mediated in I2R activation for increase of glucose uptake in the skeletal muscle cell and I2R will be a new target for diabetic therapy.