Dung-Shen Downregulates the Synergistic Apoptotic Effects of Angiotensin II Plus Leu 27-IGF II on Cardiomyoblasts.

BACKGROUND: Insulin growth factor II (IGFII) is expressed after ischemic stress in pig hearts and after myocardial infarction in humans. However, its receptor (IGFIIR) cannot be found in normal adult hearts. Moreover, a mouse IGFII overexpression model showed a heart and kidney hypertrophy phenomenon similar to Beckwith-Wiedemann syndrome in humans. The previous studies from our lab showed that an increase in AngII in H9c2 cells causes an elevation in IGFII and IGFIIR through MEK and JNK activation, leading to a rise in intracellular Ca(2+) ions, activation of calcineurin by PLC-ß3 via Gαq, insertion into mitochondrial membranes of BAD, and apoptosis via activation of caspases 9 and 3. Codonopsis pilosula (Dung-shen) has various uses in traditional Chinese medicine, including lowering blood pressure, and increasing red and white blood cell counts. METHODS: The purpose of our study is to investigate whether the addition of C. pilosula will attenuate the AngII plus Leu27-IGFII-induced apoptosis in H9c2 cardiomyoblast cells. RESULTS: From MTT [3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-tetrazolium bromide] results, it was revealed that AngII plus Leu(27)-IGFII significantly reduced cell viability, which was reversed by C. pilosula. Additionally, C. pilosula also reversed apoptosis (TUNEL staining) increased by AngII plus Leu27-IGFII. Up-regulation of caspase 3 by AngII plus Leu27-IGFII was attenuated by C. pilosula treatment, as shown in western blotting assay and immunofluorescence microscopy results. CONCLUSIONS: C. pilosula is able to suppress the apoptotic pathway enhanced by AngII plus Leu27-IGFII in myocardial cells. KEY WORDS: Angiotensin II; Apoptosis; Codonopsis pilosula; Leucine27-insulin like growth factor II; Mitochondrial outer membrane permeability.

MALDI-TOF mass spectrometry analysis of small molecular weight compounds (under 10 KDa) as biomarkers of rat hearts undergoing arecoline challenge.

CONTEXT: Statistical and clinical reports indicate that betel nut chewing is strongly associated with progression of oral cancer because some ingredients in betel nuts are potential cancer promoters, especially arecoline. Early diagnosis for cancer biomarkers is the best strategy for prevention of cancer progression. Several methods are suggested for investigating cancer biomarkers. Among these methods, gel-based proteomics approach is the most powerful and recommended tool for investigating biomarkers due to its high-throughput. However, this proteomics approach is not suitable for screening biomarkers with molecular weight under 10 KDa because of the characteristics of gel electrophoresis. OBJECTIVE: This study investigated biomarkers with molecular weight under 10 KDa in rats with arecoline challenge. MATERIALS AND METHODS: The centrifuging vials with membrane (10 KDa molecular weight cut-off) played a crucial role in this study. After centrifuging, the filtrate (containing compounds with molecular weight under 10 KDa) was collected and spotted on a sample plate for MALDI-TOF mass spectrometry analysis. RESULTS: Compared to control, three extra peaks (m/z values were 1553.1611, 1668.2097 and 1740.1832, respectively) were found in sera and two extra peaks were found in heart tissue samples (408.9719 and 524.9961, respectively). These small compounds should play important roles and may be potential biomarker candidates in rats with arecoline. DISCUSSION AND CONCLUSIONS: This study successfully reports a mass-based method for investigating biomarker candidates with small molecular weight in different types of sample (including serum and tissue). In addition, this reported method is more time-efficient (1 working day) than gel-based proteomics approach (5~7 working days).

Histone acetylation is essential for ANG-II-induced IGF-IIR gene expression in H9c2 cardiomyoblast cells and pathologically hypertensive rat heart.

The IGF-II/mannose 6-phosphate receptor (IGF-IIR/Man-6-P) up-regulation correlates with heart disease progression and its signaling cascades directly trigger pathological cardiac hypertrophy, fibrosis, and cardiomyocytes apoptosis. IGF-IIR gene expression/ suppression is able to prevent myocardial remodeling. However, the regulating mechanisms for the IGF-IIR gene remain unclear. This study performed reverse transcriptase PCR (RT-PCR) and methylation-specific PCR (MS-PCR) to detect expression and DNA methylation of CpG islands within the IGF-IIR genomic DNA region. Our finding revealed that the IGF-IIR gene was up-regulated both in H9c2 cells treated with tumor necrosis factor-alpha (TNF-α), lipopolysaccharide (LPS), angiotensin II (ANGII) and inomycin, and age-dependently in spontaneously hypertensive rat (SHR) heart. For the DNA methylation study, although there were four CpG islands within IGF-IIR genomic regions, the DNA methylation distribution showed no change either in cells treated with ANGII or in the SHR heart. Using chemical inhibitors to individually block histone acetyltransferase (HAT) and histone deacetylase (HDAC) activity, we found that histone acetylation was essential for ANGII-induced IGF-IIR gene expression using RT-PCR and luciferase assay. The Chromatin immuno-precipitation assay indicated that acetyl-Histone H3 and acetyl-Histone H4 associated with the IGF-IIR promoter increased in the presence of ANGII, otherwise methyl-CpG binding domain protein 2 (MeCP2) is disassociated with this. Taken together, this study demonstrates that histone acetylation plays a critical role in IGF-IIR up-regulation during pathological cardiac diseases and might provide a targeting gene in transcriptional therapies for the failing heart.

Pro-inflammation and pro-fibrosis factors were highly induction in heart tissues of carotid arteries balloon-injured animal model.

To investigate the changes of cardiomyocyte inflammation and fibrosis factors in heart of carotid artery balloon injury inflammatory rat model. Using rat carotid artery balloon injury model to detect left ventricular characteristics at 2 h, 2 days and 14 days after surgery using hematoxylin-eosin (H&E) gross stain, Masson's trichome stain and Western blot analysis for inflammatory and fibrosis-induced factors, tumour necrosis factor α (TNFα), JNK1, P38α, connective tissue growth factor (CTGF), SP1 and transforming growth factor ß (TGFß) protein expressions. The rat carotid arteries were injured after 2 h, 2 days and 14 days. Balloon-angioplasty to H&E stain results showed the increasing trend of left ventricular wall at 2 h and 2 days; then, the left ventricular wall became thinner, and the left ventricular chamber became enlarged and dilated after 14 days of carotid artery balloon injury. In addition, the Masson's trichome stain results showed that the left ventricular section has fibrosis-related blue staining (collagen) at 2 and 14 days after rat carotid artery balloon injury, and became even more severe at 14 days. Furthermore, we observed the protein expression level changs, which include TNFα, JNK1, P38α, CTGF, SP1 and TGFß using Western blotting assay. All proteins were induced at 2 h, 2 days and then reached the maximal level at 14 days. The vessel inflammation was associated with cardiac inflammatory and fibrosis effects during or after carotid artery balloon injury.

Anti-inflammatory effects of probiotic Lactobacillus paracasi on ventricles of BALB/C mice treated with ovalbumin.

Lactic acid bacteria (LAB) are microorganisms that benefit animals with allergic diseases and intestinal disorders such as inflammatory bowel disease. We propose that LAB can prevent cardiomyocytes inflammation and apoptosis in BALB/c mice using an ovalbumin (OVA)-induced allergy. Thirty-nine male BALB/c mice were divided into five groups: normal control, allergy control and three allergy groups each treated with Kefir I (Kefir I), Kefir II (Kefir II) or GM080 products (GM080). The myocardial architecture and apoptotic molecules in the excised left ventricle from these mice were investigated and post-treatment effects were evaluated. The inflammatory pathway, including toll-like receptor 4 (TLR4), phospholate-Jun-N-terminal kinase (p-JNK), JNK1/2 and tumor necrosis factor- alpha (TNF-α) and the mitochondria-dependent apoptosis phospholate-p38 (p-p38), Bcl-2 associated agonist of cell death (Bad), Bcl-2 associated X (Bax) and activated caspase 3, were found to be significant- ly increased in the hearts of allergy mice. The expression of phospholate-nuclear factor-κB (p-NFκB), TNF-α, p-p38 and Bad protein products were reduced or retarded in the Kefir I- or II-treated allergy group. The GM080-treated allergy group exhibited significantly lower p-JNK, JNK1/2, phospholate- Ikappa B (p-IκB), Bax and Bad protein products than the Kefir I and Kefir II allergy groups. These results indicate that LAB can reduce inflammation and prevent apoptosis of cardiomyocytes in the heart of OVA-induced allergy mice.

Isolated septic discitis associated with Streptococcus bovis bacteremia.

Streptococcus bovis is a Gram-positive coccus that can be found in the intestinal flora of healthy people; it is also associated with colon cancer and infective endocarditis. We report on a 79-year-old male who initially presented with acute-onset lower back pain. Streptococcus bovis was detected in repeated blood cultures, and magnetic resonance imaging of the lumbar spine revealed septic discitis of the L2-L3 intervertebral disc. Excision and debridement of the intervertebral disc was performed and a tissue culture tested positive for S. bovis. Repeat echocardiography and colonoscopy showed no signs of vegetation or tumor lesions, respectively. We diagnosed the patient with isolated septic discitis caused by S. bovis-induced bacteremia. The patient was discharged after six weeks of antibiotic therapy.

Lipopolysaccharide upregulates uPA, MMP-2 and MMP-9 via ERK1/2 signaling in H9c2 cardiomyoblast cells.

Upregulation of urokinase plasminogen activator (uPA), tissue plasminogen activator (tPA), and matrix metallopeptidases (MMPs) is associated with the development of myocardial infarction (MI), dilated cardiomyopathy, cardiac fibrosis, and heart failure (HF). Evidences suggest that lipopolysaccharide (LPS) participates in the inflammatory response in the cardiovascular system; however, it is unknown if LPS is sufficient to upregulate expressions and/or activity of uPA, tPA, MMP-2, and MMP-9 in myocardial cells. In this study, we treated H9c2 cardiomyoblasts with LPS to explore whether LPS upregulates uPA, tPA, MMP-2, and MMP-9, and further to identify the precise molecular and cellular mechanisms behind this upregulatory responses. Here, we show that LPS challenge increased the protein levels of uPA, MMP-2 and MMP-9, and induced the activity of MMP-2 and MMP-9 in H9c2 cardiomyoblasts. However, LPS showed no effects on the expression of tissue inhibitor of metalloproteinase-1, -2, -3, and -4 (TIMP-1, -2, -3, and -4). After administration of inhibitors including U0126 (ERK1/2 inhibitor), SB203580 (p38 MAPK inhibitor), SP600125 (JNK1/2 inhibitor), CsA (calcineurin inhibitor), and QNZ (NFkappaB inhibitor), the LPS-upregulated expression and/or activity of uPA, MMP-2, and MMP-9 in H9c2 cardiomyoblasts are markedly inhibited only by ERK1/2 inhibitors, U0126. Collectively, these results suggest that LPS upregulates the expression and/or activity of uPA, MMP-2, and MMP-9 through ERK1/2 signaling pathway in H9c2 cardiomyoblasts. Our findings further provide a link between the LPS-induced cardiac dysfunction and the ERK1/2 signaling pathway that mediates the upregulation of uPA, MMP-2 and MMP-9.

Down regulation of IGF-I and IGF-IR gene expression in right atria tissue of ventricular septal defect infants with right atria hypoxemia.

BACKGROUND: Our previous studies showed serum insulin-like growth factor-I (IGF-I) concentrations significantly decreased in infants with congenital ventricular septal defect (VSD) and that they were associated with increased concentrations of growth hormone. In order to confirm the relationship between IGF-I axis and VSD, we further compared the IGF-I and insulin-like growth factor-I receptor(IGF-IR) gene expressions in the cardiac tissue of VSD infants. METHODS: Right atrium biopsies of 27 infants were studied. Five infants not having VSD were classified as controls (Group I). Twenty VSD patients were then divided into 2 groups according to their shunting magnitude index (level of pulmonary vascular resistance compared with systemic vascular resistance, Qp/Qs). VSD patients with minor shunts (Qp/Qs<1.7) were classified as Group II; VSD patients with larger shunts (Qp/Qs> or =2) as Group III. Besides, seven tetralogy of fallot (TOF) with shunt (Qp/Qs>4) infants were classified as the Group IV. A non-radioactive DIG-RNA probe detection system, western blotting and immunohistochemistry were used to detect the gene expression levels and protein products of IGF-I and IGF-IR in the right atrium samples of VSD infants. RESULTS: The relative protein levels of IGF-I were 0.96+/-0.05, 0.43+/-0.03, 0.15+/-0.04, 0.12+/-0.03 and IGF-IR were 0.80+/-0.08, 0.57+/-0.03, 0.38+/-0.02, 0.24+/-0.04 in the right atrium of 4 group patients. The relative mRNA levels of IGF-I were 0.95+/-0.01, 0.41+/-0.03, 0.29+/-0.05, 0.15+/-0.01 and IGF-IR were 0.85+/-0.05, 0.56+/-0.03, 0.17+/-0.01, 0.18+/-0.01, respectively. There was a significantly greater but more gradual decrease in protein levels and in mRNA levels of IGF-I and IGF-IR in Group II (p<0.05), Group III and IV (p<0.01) than in Group I. The results of immunohistochemistry also demonstrated a similar decrease in VSD patients. In addition, the decrease of mRNA and protein levels in IGF-I/IGF-IR of VSD patients show related to the saturation of oxygen in the right atrium and the ratio of systolic right ventricular pressure to left ventricular pressure. CONCLUSION: We further confirmed the down regulation of IGF-I/IGF-IR in cardiac tissue of VSD infants and the decrease to be associated with shunt magnitude and the severity of hypoxemia in the cardiac chamber of VSD.

Homocysteine and other biochemical parameters in Type 2 diabetes mellitus with different diabetic duration or diabetic retinopathy.

BACKGROUND: Cardiovascular risk-related markers in type 2 diabetes mellitus (DM) have not been well understood. METHODS: Serum and urine samples for biochemical and immunologic analysis were collected from 204 normal subjects and 257 type 2 DM patients, the latter of which were further classified by different diabetic duration with or without retinopathy. RESULTS: Glycosylated hemoglobin A1c, triglyceride, lipase, free fatty acid, albumin creatinine ratio (ACR), lactate dehydrogenase (LDH) and homocysteine were significantly increased in DM patients, whereas high density lipoprotein cholesterol and bilirubin were significantly decreased in DM patients, compared with normal subjects. Lipid profiles, ACR, bilirubin, uric acid, creatine kinase, and hsCRP were not changed in DM patients with different diabetic duration or diabetic retinopathy. Lactate dehydrogenase in DM patients with duration >20 years and homocysteine in patients with duration >10 years was significantly higher than those with duration <5 years. Homocysteine was significantly increased in DM patients with retinopathy, compared with DM patients without retinopathy. CONCLUSION: The increased triglyceride, lipase, free fatty acid, albumin creatinine ratio, lactate dehydrogenase and homocysteine as well as decreased high density lipoprotein cholesterol and anti-oxidative bilirubin in DM patients should be cautious and considered as risks for increasing DM complication. Homocysteine might be associated with longer diabetic duration and microvascular complication of retinopathy in diabetes.

Long-term efficacy and safety of very-low-dose amiodarone treatment for the maintenance of sinus rhythm in patients with chronic atrial fibrillation after successful direct-current cardioversion.

BACKGROUND: Long-term maintenance of sinus rhythm after successful conversion of chronic atrial fibrillation (CAF), often ameliorates patients' symptoms, reduces the risk of ischemic stroke and improves cardiovascular hemodynamics. This prospective study aims to evaluate the long-term efficacy and safety of very low-dose amiodarone (100 mg daily) for the maintenance of sinus rhythm after successful direct-current (DC) cardioversion in patients with CAF and rheumatic heart disease (RHD) post intervention. METHODS: This study was a randomized prospective trial. One day after successful DC cardioversion (remained normal sinus rhythm) in patients with CAF and RHD post intervention for more than six months and adequate anticoagulation, all were randomly administered either amiodarone 200 mg daily in group A or amiodarone 100 mg daily in group B. RESULTS: A total of 76 patients (40 men and 36 women) were examined from February 1998 to December 1999. The mean age of the patients was (66 +/- 10) years, and the mean follow-up was (67 +/- 8) months (range 61 to 84 months). Actuarial rates of the maintenance of sinus rhythm were similar in the two groups after 5 years of follow-up. Four patients (11%) in group A but none in group B experienced significant adverse effects that necessitated withdrawal of amiodarone. No death occurred during the study period. CONCLUSION: A very low dose of amiodarone results in adequate long-term efficacy and is safe for maintaining sinus rhythm in patients with CAF and RHD post intervention after successful DC cardioversion.

Serum MMP-9 activity as a diagnosing marker for the developing heart failure of post MI patients.

Myocardial infarction (MI) is the result, in mostly cases, of the destabilization and rupture of atherosclerotic lesions. The destruction of cardiac tissue resulting from myocardial ischemia could further result in heart failure. It has been suggested that plaque instability may be mediated by matrix metalloproteinase (MMP) family. Studies have identified increased MMP-2 and MMP-9 in human platelets, and acute myocardial infarction patients with elevated MMP-2 and MMP-9 levels. However, the alteration of MMP-2 and MMP-9 from post MI left ventricle remodeling to heart failure remains to be clarified. The purpose of this study is to investigate the serum concentrations and activities of MMP-2 and MMP-9 in the developing heart failure from post MI patients. Twenty eight patients with MI without heart failure (Killip FC I) (group A; compensated) and twenty seven MI patients with heart failure (Killip II-III) (group B; decompensated) were collected to evaluate the serum levels and activities of MMP-2 and MMP-9 by ELISA and Zymography, respectively. It was observed that the both serum levels and activities of MMP-9 significantly increased (P < 0.01) in decompensated group compared to compensated group, but there was no significant difference of serum MMP-2 levels and activities between two groups. The highly elevated serum MMP-9 concentration of decompensated patients is not related with inflammatory or localized infarct area of myocardium and the real mechanisms remain to be revealed. We suggest that the increase of MMP-9 levels and activity may be used as a new marker to diagnose the development of heart failure in patients with post MI, and provide the therapeutic implications in the future.

ALDH1A3, the Major Aldehyde Dehydrogenase Isoform in Human Cholangiocarcinoma Cells, Affects Prognosis and Gemcitabine Resistance in Cholangiocarcinoma Patients.

PURPOSE: Intrahepatic cholangiocarcinoma is a fatal primary liver cancer resulting from diagnosis at an advanced stage. Understanding the mechanisms of drug resistance and metastasis of cholangiocarcinoma may improve the disease prognosis. Enhanced aldehyde dehydrogenase (ALDH) activity is suggested to be associated with increased drug resistance and the metastasis. This study aims to investigate the roles of the ALDH isoforms in cholangiocarcinoma. EXPERIMENTAL DESIGN: Aldefluor assays, RT-PCR, and Western blot analysis were used to identify the major ALDH isoforms contributing to Aldefluor activity in human cholangiocarcinoma cell lines. We manipulated isoform expression in HuCCT1 cells to elucidate the role of ALDH1A3 in the malignant progression of these cells. Finally, we used immunohistochemical staining to evaluate the clinical significance of ALDH1A3 in 77 hepatectomized cholangiocarcinoma patients and an additional 31 patients with advanced cholangiocarcinoma who received gemcitabine-based therapy. RESULTS: ALDH(high) cholangiocarcinoma cells not only migrated faster but were more resistant to gemcitabine. Among the 19 ALDH isoforms studied, ALDH1A3 was found to be the main contributor to Aldefluor activity. In addition, we also found that knockdown of ALDH1A3 expression in HuCCT1 cells markedly reduced not only their sensitivity to gemcitabine, which might be attributed to a decreased expression of ribonucleotide reductase M1, but also their migration. Most importantly, this enzyme was also identified as an independent poor prognostic factor for patients with intrahepatic cholangiocarcinoma, as well as a prognostic biomarker of gemcitabine-treated patients. CONCLUSIONS: ALDH1A3 plays an important role in enhancing malignant behavior of cholangiocarcinoma and serves as a new therapeutic target. Clin Cancer Res; 22(16); 4225-35. ©2016 AACR.

Second-hand smoke-induced cardiac fibrosis is related to the Fas death receptor apoptotic pathway without mitochondria-dependent pathway involvement in rats.

Exposure to environmental tobacco smoke has been epidemiologically linked to heart disease among nonsmokers. However, the molecular mechanism behind the pathogenesis of cardiac disease is unknown. In this study, we found that Wistar rats, exposed to tobacco cigarette smoke at doses of 5, 10, or 15 cigarettes for 30 min twice a day for 1 month, had a dose-dependently reduced heart weight to body weight ratio and enhanced interstitial fibrosis as identified by histopathologic analysis. The mRNA and activity of matrix metalloprotease-2 (MMP-2), representing the progress of cardiac remodeling, were also elevated in the heart. In addition, we used reverse-transcriptase polymerase chain reaction and Western blotting to demonstrate significantly increased levels of the apoptotic effecter caspase-3 in treated animal hearts. Dose-dependently elevated mRNA and protein levels of Fas, and promoted apoptotic initiator caspase-8 (active form), a molecule of a death-receptor-dependent pathway, coupled with unaltered or decreased levels of cytosolic cytochrome c and the apoptotic initiator caspase-9 (active form), molecules of mitochondria-dependent pathways, may be indicative of cardiac apoptosis, which is Fas death-receptor apoptotic-signaling dependent, but not mitochondria pathway dependent in rats exposed to second-hand smoke (SHS). With regard to the regulation of survival pathway, using dot blotting, we found cardiac insulin-like growth factor-1 (IGF-1) and IGF-1 receptor mRNA levels to be significantly increased, indicating that compensative effects of IGF-1 survival signaling could occur. In conclusion, we found that the effects of SHS on cardiomyocyte are mediated by the Fas death-receptor-dependent apoptotic pathway and might be related to the epidemiologic incidence of cardiac disease of SHS-exposed nonsmokers.

17beta-estradiol reduces cardiac hypertrophy mediated through the up-regulation of PI3K/Akt and the suppression of calcineurin/NF-AT3 signaling pathways in rats.

This study was designed to determine the effects of 17beta-estradiol (E2) in overcoming the cardiac over-loading and cardiac fibrosis in rats. E2 (100 ng/kg) or oil was applied in female Sprague-Dawley rats with or without bilateral ovariectomy and with or without coarctation of the abdominal aorta after 4 or 8 days. By post-operative day 4, the heart weight, the left ventricular weight, the latent form of MMP-2 in rat hearts with or without the ovary intact had significantly increased while these changes were reversed after E2 treatment. Although animals with the ovaries intact overcame the hypertrophic effects and the consumption of MMP-2, these effects were not restored in ovariectomized animals in which more fibrosis could be found by day 8. Among the IGF-I signaling, the levels of IGF-I, the activities of PI3K-Akt for cardiomyocyte survival, and MEK-ERKs for non-cardiomyocyte proliferation pathways had significantly increased by day 4. These increasing trends were enhanced by E2 treatment. However, down-regulation was only observed on day 8 in ovariectomized animals. Similarly, elevated expressions of the steady-state mRNA of IGF-I, IGF-IR, and Cox vb were observed on day 4 in animals with the ovaries intact and these expressions were enhanced by E2 treatment. In contrast, down-regulation on day 8 in ovariectomized animals was not enhanced by E2. The calcineurin/NFAT-3 pathway was suppressed on day 4 but was elevated on day 8 in ovariectomized animals. These findings indicate that signaling pathways may be plausible mechanisms for the cardiac protective effects of E2 administration.

The novel heterocyclic trioxirane [(1,3,5-tris oxiran-2-yl)methyl)-1,3,5-triazinane-2,4,6-trione (TATT)] exhibits a better anticancer effect than platinum-based chemotherapy by induction of apoptosis and curcumin further enhances its chemosensitivity.

The heterocyclic trioxirane compound [1,3,5-tris((oxiran-2-yl)methyl)-1,3,5-triazinane-2,4,6-trione (TATT)] is a synthetic compound which has been used as an experimental anticancer agent in human clinical trials. Curcumin, an active natural compound in turmeric and curry, is an ingredient commonly used in the traditional diet of many Asian countries. In the present study, we observed that TATT exhibited a better anticancer effect on chemoresistant human colorectal cancer HT-29 cells and displayed less cytotoxicity on normal human umbilical vein endothelial cells, compared with FDA-approved anticancer drugs (cisplatin, carboplatin, or oxaliplatin) using MTT assay. TATT also induced a stronger apoptotic effect than that seen with the three studied anticancer drugs, as characterized by externalization of phosphatidylserine using flow cytometry. Administration of caspase 8-specific inhibitor (z-IETD-fmk) and mitochondrial permeability transition pore inhibitor (cyclosporin A) demonstrated that TATT-induced apoptosis proceeded via both extrinsic and intrinsic signaling pathways. It is noteworthy that coadministration of curcumin further significantly increased TATT-induced cytotoxicity, externalization of phosphatidylserine (representing early apoptosis), and the percentages of cells at the sub-G1 phase (representing late apoptosis), producing an additivity and/or synergistic effect, and vice versa. Suppression of nuclear NF-κB was involved in curcumin-enhanced chemosensitivity of TATT. Overall, our data indicate that TATT exerts a chemotherapeutic effect on colorectal cancer cells and coadministration of curcumin enhances the treatment effect of TATT.

Garlic oil attenuates the cardiac apoptosis in hamster-fed with hypercholesterol diet.

Hypercholesterolemia is a well established risk factor for cardiac cell apoptosis. The purpose of this study is to evaluate the effects of garlic oil on cardiac apoptosis induced by a hypercholesterol diet. Twenty-four male Golden-Syrian hamsters at 3 months of age were randomly divided into three groups, control, cholesterol and garlic oil groups received a chow diet, chow diet with 2% cholesterol, and chow diet with 2% cholesterol and 1% garlic oil for 8 weeks, respectively. The TUNEL-positive apoptotic cells, and several apoptotic proteins were significantly induced in the excised left ventricle in cholesterol group, whereas significant reduction was observed in cholesterol plus garlic oil group. The IGFI receptor dependent survival pathway was inhibited in cholesterol group whereas it was obviously reversed in cholesterol plus garlic oil group. Our results suggest that administration of garlic oil shows protective effects on cardiac apoptosis in rats with high cholesterol intake.

Dung-shen (Codonopsis pilosula) attenuated the cardiac-impaired insulin-like growth factor II receptor pathway on myocardial cells.

Previous studies from our lab showed that increase in AngII in H9c2 cells causes elevated IGFII and IGFIIR through MEK and JNK, leading to rise in intracellular calcium, calcineurin activation by PLC-ß3 via Gαq, insertion into mitochondrial membranes of Bad, and apoptosis via caspases 9 and 3. Codonopsis pilosula is traditionally used to lower blood pressure. The purpose of our study is to investigate if C. pilosula attenuates AngII plus Leu(27)-IGFII-induced calcium influx and apoptosis in H9c2 cardiomyoblasts. C. pilosula significantly attenuated AngII induced IGFIIR promoter activity. Leu(27)-IGFII was applied to enhance the AngII effect. C. pilosula also reversed Ca(2+) influx, MOMP and apoptosis increased by AngII plus Leu(27)-IGFII. Molecular markers in IGFIIR apoptotic pathway (IGFIIR, calcineurin, etc.) and IGFIIR-Gαq association were downregulated by C. pilosula. However, p-Bad(Ser136) and Bcl-2 were increased. Therefore, C. pilosula suppresses AngII plus Leu(27)-IGFII-induced IGFII/IGFIIR pathway in myocardial cells.

The long-term effect of statins on the risk of new-onset diabetes mellitus in elderly Taiwanese patients with hypertension and dyslipidaemia: a retrospective longitudinal cohort study.

BACKGROUND: HMG-CoA reductase inhibitors (statins) have been linked to new-onset diabetes (NOD). Individual statins may differ in the extent to which they increase the risk for NOD; however, the effect of statins on the development of NOD in elderly hypertensive and dyslipidaemic patients has not been well studied. OBJECTIVE: The aim of this study was to investigate the relative risk for NOD among elderly (age ≥65 years) hypertensive and dyslipidaemic Taiwanese patients who received different statins. METHODS: This was a retrospective cohort study performed using data from claim forms provided to the central regional branch of the Bureau of National Health Insurance in Taiwan from July 2004 to December 2009. Prescriptions for statins before the index date were retrieved from a prescription database. We estimated the hazard ratios (HRs) of NOD associated with statin use. Non-diabetic subjects served as the reference group. RESULTS: A total of 2735 NOD cases were identified among 15,637 elderly hypertensive and dyslipidaemic patients during the study period. The risk of NOD after adjusting for sex, age, concomitant medication and mean dose of prescription was lower among users of atorvastatin (HR 0.77; 95% CI 0.71, 0.83) and rosuvastatin (HR 0.65; 95% CI 0.51, 0.82) than among non-users. Patients who took lovastatin (HR 1.38; 95% CI 1.26, 1.50) or simvastatin (HR 1.30; 95% CI 1.14, 1.48) were at higher risk of developing NOD than non-users. Pravastatin and fluvastatin were not associated with increased risk of NOD. CONCLUSIONS: The results of this study suggest that elderly hypertensive and dyslipidaemic patients who take atorvastatin or rosuvastatin are at lower risk of NOD. Lovastatin and simvastatin were associated with a significant increase in the risk of NOD.

Syncope with spontaneous coronary artery dissection in an elderly woman.

Diagnosis of spontaneous coronary artery dissection (SCAD) is challenging because of its rarity and uncertain etiology. It frequently occurs in young women during pregnancy and in the postpartum period, and rarely found in elder women with no history of cardiovascular disease or coronary risk factors. In this article we report a case of SCAD in a 75-year-old woman without traditional cardiovascular risk factors who presented with syncope and mild chest discomfort. There were no abnormal electrocardiographic changes and no elevated cardiac enzymes were detected. Computed tomography of brain revealed nothing abnormal. Coronary artery disease was suspected. Coronary angiogram revealed dissection in the middle left circumflex artery. The patient underwent percutaneous transluminal coronary angioplasty and was free of symptoms at 6-month follow-up. Our report suggests that emergency coronary angiography is indicated if syncope caused by coronary artery disease is suspected.

IGF-II and MMP9 as surgical repair indicators of ventricular septal defects.

OBJECTIVE: The insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-3 (IGFBP-3) and human growth hormone (h-GH) have been recognized as therapeutic targets for the heart disease therapy. The bioavailability and actions of insulin-like growth factors-II (IGF-II) and matrix metalloproteinase-9 (MMP9) are important for embryonic development and cardiomyocyte differentiation as well. However, the clinical manifestations following the change in the serum IGF-II and MMP9 in infants with isolated ventricular septal defect (VSD) undergoing surgical repair have not been clearly defined. STUDY DESIGN: Serum samples were collected from 72 infants: Twenty normal infants (group I) and 51 consecutive infants with echocardiography established isolated VSD (aged from 3 months to 1 year) were investigated. Among the 51 infants with VSD, 28 with shunt fraction, Qp/Qs < or = 1.5 were free of congestive heart failure symptoms (group II); 23 with shunt fraction, Qp/Qs > or = 2.0 were in congestive heart failure (group IIIa); and 23 of these 23 infants had undergone VSD repair 6 months before their second study (group IIIb). All insulin-like growth factors-II (IGF-II) and human growth hormone (h-GH), insulin like growth factor binding protein-3 (IGFBP-3) and its specific serum protease-MMP9 concentration were analyzed using ELISA and zymography, respectively. RESULTS: Serum IGF-II and MMP9 exhibited significant decreasing trends among the three groups and significantly lower concentrations of IGF-II, IGF-II/IGFBP-3 ratio and MMP9, were found only in the severe group whereas h-GH/IGF-II ratio became significantly higher in this group. Moreover, there were no significant differences in these parameters between the infants after surgical correction and the normal ones. CONCLUSIONS: The improvement in IGF-II and MMP9 serum concentration was identified in infants with VSD after surgical repair. These findings also indicate a significant relationship between IGF-II, MMP9 and VSD which might be used as diagnosis and prognosis indicators for this defect. Slight reductions in IGF-II/IGFBP3 ratio and slight increase in the h-GH/IGF-II ratio indicate mild VSD. The reductions in the MMP9, IGF-II, and IGF-II/IGFBP3 ratio plus high increase in the h-GH/IGF-II ratio indicate severe VSD.