Effect of L158,809 and Cilazapril on the Expression of TGF-β_1 and Secretion of Extracellular Matrix Proteins in Cultured Human Mesangial Cells

Objective: To explore the effect of L158, 809 (angiatensin Ⅱ receptorMockers, ARBs) and Cilazapril (Angiotensin converting enzyme inhibitor, ACEI) on the expression oftransforming growth factor-β_1 (TGF-β_1) and secretion of fibronectin, laminin and type Ⅳcollagen from the cultured human mesangial cells . Methods: Human mesangial cells were cultured indifferent glucose (5.6 mmol/L and 30 mmol/L) and agents (1, 10, 100 and 500 μmol/L) concentrations. The proliferation of mesangial cells were detected at 24, 48 and 72 h . Then the mesangial cellsare divided into four groups, low glucose (5.6 mmol/L) control group, high glucose (30 mmol/L)control group , L158, 809 (10 μmol/L) group and cilazapril (10 μmol/L) group. Forty- eight hourslater, the expression of TGF-β_1 were detected by RT-PCR. Concentrations of TGF-β_1 ,fibronection, laminin and type Ⅳ collagen in the su-pematants of the, mesangial cells weredetermined by EUSA and radioimmunoassay methods. Results: Compared with low glucose control group,the mesangial cells under high glucose medium show excessive proliferation and more TGF-β_1,fibronectin, laminin and type Ⅳ collagen in the supernatant. The expression of TGF-β_1 mRNA wasalso significantly increased under high glucose. The levels of TGF-β_1 and ECM (extracellularmatrix) proteins in the L158, 809 group and cilazapril group are obviously lower than that of thehigh glucose control group. The expression of TGF-β_1 mRNA was markedly decreased in the L158, 809group and cilazapril group compared with that of high glucose control group . Conclusion: Highglucose stimulated the cultured human mesangial cells to excessively proliferate, express TGF-β_1and secrete ECM proteins, and the high glucose-indeced changes were suppressed by either L158, 809and cilazapril.

SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway

While severe acute respiratory syndrome coronavirus （SARS-CoV）~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 （ACE2）, the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research.

Cardiovascular and cerebrovascular outcomes in elderly hypertensive patients treated with either ARB or ACEI

Background Although angiotensin converting enzyme inhibitors （ACEI） and angiotensin receptor blockers （ARB） are equally important in the treatment of hypertension, there is less evidence whether they have equal cardiovascular and cerebrovascular protective effects, especially in elder hypertensive patients. This study aims to clarify this unresolved issue. Methods This cross-sectional study included clinical data on 933 aged male patients with hypertension who received either an ARB or ACEI for more than two months between January 2007 and May 2011. The primary outcome was the composite of cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke. The secondary endpoints were unstable angina, new atrial fibrillation, and transient ischemic attack. Results The median follow-up time was 24 months. Age, drug types, cerebral infarction history, renal dysfunction history were the independent predictors of the primary endpoint. The risk of an occurrence of a primary endpoint event was higher in the ARB group than the ACEI group [P = 0.037, hazard ratios （HR）： 2.124, 95% confidence interval （95% CI）： 1.048-4.306]. The Kaplan-Meier method also suggests that the rate of primary endpoint occurrence was higher in the ARB group than the ACEI group （P = 0.04）. In regard to the secondary endpoints, there were no significant differences between the two treatment arms （P = 0.137, HR： 1.454, 95% CI： 0.888-2.380）. Patient age and coronary heart disease history were independent predictors of the secondary endpoint. Conclusion ACEI were more effective than ARB in reducing cardiovascular and cerebrovascular morbidity and mortality in aged patients with hypertension.

Inappropriate use of digoxin in patients presenting with digoxin toxicity

Background Digoxin remains widely used today despite its narrow therapeutic index and toxicity. The objective of this study was to investigate the percentage of inappropriate use of digoxin and long-term outcomes of elderly patients hospitalized for digoxin toxicity. Methods The study included 99 consecutive patients hospitalized for digoxin toxicity. The other study criteria for the inappropriate use of digoxin was regarded if participants having depressed left ventricular systolic function （ejection fraction 〈 45%） who were not on optimal medical therapy including beta-blocker and angiotensin-converting-enzyme inhibitor therapy or if participants having permanent AF who were not on optimal beta-blocker therapy. Results Appropriate digoxin usage was confirmed in 33 of patients in spite of its narrow therapeutic index. A total of 16 of 99 patients died, with a mean follow-up time of 22.1 ± 10.3 months. Conclusions Contrary to popular belief, the rate of inappropriate digoxin usage remains high. On account of its narrow therapeutic index and toxicity, digoxin should be used more carefully according to the current evidence and guidelines.

Purification and Characterization of Angiotensin I Converting Enzyme Inhibition Peptides from Sandworm Sipunculus nudus

Three angiotensin I converting enzyme(ACE) inhibition peptides were isolated from sandworm Sipunculus nudus protein hydrolysate prepared using protamex. Consecutive purification methods, including size exclusion chromatography and reverse-phase high performance liquid chromatography(RP-HPLC), were used to isolate the ACE inhibition peptides. The amino acid sequences of the peptides were identified as Ile-Asn-Asp, Val-Glu-Pro-Gly and Leu-Ala-Asp-Glu-Phe. The IC_(50) values of the purified peptides for ACE inhibition activity were 34.72 μmol L^(-1), 20.55 μmol L^(-1) and 22.77 μmol L^(-1), respectively. These results suggested that S. nudus proteins contain specific peptides that can be released by enzymatic hydrolysis. This study may provide an experimental basis for further systematic research, rational development and clinical utilization of sandworm resources.

The impact of optimal medical therapy at discharge on mortality in patients with coronary artery disease

Objective To analyze the current usage of optimal medical therapy （OMT）, influencing factors, and the predictive value of OMT for all-cause mortality in coronary artery disease （CAD） patients with different subgroups. Methods A total of 3176 CAD patients confirmed by coronary angiography were included. OMT was defined as the combination of anti-platelet drugs, statins, beta blockers, and angiotensin converting enzyme inhibitors or angiotensin receptor blockers. Factors for OMT and its prognostic value were analyzed in CAD patients across different subgroups. Results Out of 3176 patients, only 39.8% （n = 1265） were on OMT at discharge. Factors associated with OMT at discharge were pre-admission OMT and discharge department. All-cause mortality occurred in 6.8% （n = 217） of patients. Multivariate analyses indicated that OMT was significantly associated with reduced all-cause mortality （HR： 0.65, 95% CI： 0.45~0.95; P = 0.025）. Sub-group analyses indicate that male acute coronary syndrome （ACS） patients were more likely to receive survival benefits with OMT at discharge. The positive impact of OMT at discharge was more apparent after 24 months, regardless of revascularization therapy. Four-drug combination of OMT was superior to 3-drug combination therapy in ACS patients but not in stable patients. Conclusions OMT was asso- ciated with significant improvement in survival in patients with CAD. The positive impact of OMT was distinct in the CAD patients with different characteristics.

Pharmarcogenetic Mechanism of ACE liD Polymorphism Adversely Responding to ACE Inhibitors in Regulating the ACE Promoter Activity in Neurons

The ACE （angiotensin converting enzyme） inhibitors are not only drugs widely prescribed drugs in cardiovascular diseases, but also potentially therapeutic agents in dementia. Based on the findings that the ACE inhibitors could activate the c-Jun N-terminal kinase signal to increase the ACE gene expression and that the Alu element of the human ACE gene involved in regulating ACE promoter activity, we aimed to investigate whether there are different pharmacogenetic responses of ACE I/D polymorphism to the ACE inhibitors in neurons. The three reporter vectors, pACEpro（0-SEAP, p-I-ACEpro-SEAP, and p-D-ACEpro-SEAP were used to examine the transcriptional activity of the vectors responding to the lisinopril treatment using a transient-transfection method in SH-SY5Y cells. Our results showed that lisinopril increased the promoter activity of an ACE gene by 16.7%. Additionally, we found the lisinopril enhanced the ACE promoter activity of the I-form vector by 17.2%, but adversely reduced that of the D-form vector by 16.8%, as compared with the respective control without the lisinopril treatment. Firstly, our findings had proved that the UD polymorphism of ACE gene contrarily responds to the ACE inhibitors in regulating the ACE expression in neurons, which provide a novel insight suggesting genetic testing to tailor the treatment regimens in AD （Alzheimer＇s disease） patients.

Left ventricular hypertrophy in relation to systolic blood pressure and the angiotensin converting enzyme I/D polymorphism in Chinese

Objective There is little population-based data on the prevalence and the environmental or genetic determinants of left ventricular hypertrophy （LVH） in China. The purpose of this paper is to study LVH in relation to systolic blood pressure and the angiotensin converting enzyme （ACE） insertion/deletion（I/D） polymorphism in Chinese. Methods We recorded 12- lead ECG （CardioSoft, v4.2） in 1365 residents in the Jingning County, Zhejiang Province, China. LVH was defined according to the gender-specific Sokolow-Lyon and Cornell product ECG criteria. Results Regardless of whether the Sokolow-Lyon or Cornell product ECG criteria was used, the prevalence of LVH （20.7% and 4.8%, respectively） significantly （P〈0.0001） increased with male gender （odds ratio [OR] 2.33 and 7.15） and systolic blood pressure （per 10 mm Hg increase, OR 1.46 and 1.33）. If the Sokolow-Lyon criteria was used, the prevalence of LVH was also influenced by alcohol intake （OR 1.44, P=-0.03） and body mass index （OR 0.83, P=0.0005）. The association between the Sokolow-Lyon voltage amplitude and the ACE I/D polymorphism was dependent on antihypertensive therapy （P=0.01）. In 1262 untreated subjects, but not 103 patients on antihypertensive medication, the ACE DD compared with II subjects had significantly higher Sokolow-Lyon voltage amplitudes （29.8：-0.6 vs. 28.0-3：0.5 mV, P=-0.02） and higher risk of LVH （OR 1.74, 95% CI： 1.12-2.69, P=-0.01）. Conclusion LVH is prevalent in Chinese, and is associated with systolic blood pressure and the ACE D allele. The genetic association might be modulated by antihypertensive therapy（J Geriatr Cardio12009; 6：131-136）.

Nursing care in old patients with heart failure： current status and future perspectives

The prognosis of heart failure （HF） patients has significantly improved in the last 20 years, given the advent of therapies such as angiotensin-converting enzyme （ACE） inhibitors/angiotensin receptor blockers （ARBs）, beta-block- ers, and implantable cardioverter-defibrillator （ICD）/cardiac resynchronization therapy devices （CRT）. In addition to these promising therapies, proper nursing-care is also important. Here we summarize recent progress of nursing-care strategies in older HF patients, including routine nursing care, transitional care model, self-care, and role of exercise training in old patients with HF.

A population-based case-crossover study of polyethylene glycol use and acute renal failure risk in the elderly

AIM:To evaluate the possibility of an association between polyethylene glycol(PEG) and acute renal failure(ARF) in elderly patients using a health insurance claims database.METHODS:We conducted a population-based casecrossover study using information obtained from Korean Health Insurance Review and Assessment Service(HIRA) claims from January 1,2005 to December 31,2005(Seoul,Korea).The study population consisted of elderly patients who received PEG prior to experiencing their first ARF-related hospitalization from April 1,2005 to December 31,2005.For each patient,one case and two control periods were matched.PEG use in a 2-or 4-wk window period prior to hospitalization for ARF was compared with PEG use in two earlier 2-or 4-wk control window periods.Conditional logistic regression analysis was used to estimate odds ratios(ORs) and 95% CI,adjusting for concomitant uses of diuretics,angiotensin converting enzyme inhibitors,non-steroidal anti-inflammatory drugs,antibiotics,anti-cancer drugs,and contrast media.RESULTS:Within the HIRA database which contained 1 093 262 elderly patients,1156 hospitalized ARF cases were identified.Among these cases,PEG was prescribed to 17(1.5%) patients before hospitalization.The adjusted ORs when applying the 2-and 4-wk window periods were 0.4(95% CI:0.03-5.24) and 2.1(95% CI:0.16-27.78),respectively.CONCLUSION:No increased risk of ARF was found in elderly PEG users.However,based on the limited number of study subjects,further analysis should be performed to confirm these results.

Effect of captopril or verapamil on intracellular sodium in cultured vascular smooth muscle cells

The effects of captopril (Cap) and verapamil (Ver)alone and in combination on intracellular Na+ concentration ([Na+]i) in cultured aortic smooth muscle cells (ASMC) of rabbits was evaluated by a direct measurement of [Na+]i with fluorescent dye sodium-binding benzofuran isophthalate (SBFI) combined with digital image. [Na+]i in resting cells was found to be 11.9 ± 0. 7 mmol/L. Angiotensin II (Ang-II, 0.1-10μmol/L) induced an increase of [Na+]i in concentration-dependent manner. Ver (0.1-10μmol/L) inhibited Ang-II (1 μmol/L)-induced increase in [Na+]i, while Cap enhanced Ang-II-induced increase in [Na+]i at 10μmol/L but not at 0.1-1μmol/L. Ver (0.1-1μmol/L)abolished enhancement of Ang-II-induced increase in [Na+]i by Cap. Thus, the inhibition of Capenhanced [Na+]i by Ver may provide a new hypothesis for the underlying molecular mechanism of synergistic effect of the combination of Ca2+ antagonists and angiotensinconverting enzyme inhibitors in controlling blood pressure.

The effects of acetaminophen combined with radiation on the radiosensitivity of irradiated human glioma cell progeny

Objective： To study the effects of acetaminophen （ACE） combined with radiation on the progeny of the human glioma cell line SHG-44, and to investigate if ACE may be an useful therapeutic radiosensitivity agent in the treatment of recurrent human glioma. Methods： A randomized, controlled experiment, was performed at the Department of Radiology Laboratory, the First Hospital Affiliated to Soochow University, between September 2004 and January 2006. Brain glioma SHG-44 cells were divided into three groups： SHG-44, SHG-44-10, and SHG-44-10 ＋ ACE cells groups. The SHG-44-10 cells group was irradiated with dose of 10 Gy by a linear accelerator （6 MVX）. It was passaged for 15 generations and cultured in RPMI-1640 culture media. Then SHG-44-10 ＋ ACE cells group was treated with ACE. Measures： Community re-double time, mean lethal dose （DO）, extrapolation number （N）, fraction surviving fraction irradiated by 2 Gy dose （SF2）, quasi-threshold dose （Dq）, and cell cycle. Results： The SF2 of the SHG-44, SHG-44-10, and SHG-44-10 ＋ ACE cells groups were 70.8%, 80.6% and 45.2%, respectively, with significance （P = 0.040）. The SHG-44-10 and SHG-44-10 ＋ ACE cells groups were irradiated with 8 Gy. After 12 hours, the G2/M ratio of the SHG-44-10 and SHG-44-10 ＋ ACE cells groups were indicating significantly higher ratio compared to pre-irradiated groups （P 〈 0.01）. After 24 hours, the G2/M ratio of the SHG-44-10 cells group decreased rapidly, while the ratio of the SHG-44-10 ＋ ACE cells group still maintained in high level. Conclusion： In the present study, Subtoxic dose of ACE increased the radiosensitivity of the progeny of irradiated human glioma cell. ACE may be an useful radiosensitivity agent in the treatment of recrudescent human malignant glioma.

CLINICAL OUTCOMES OF FIVE-YEAR FOLLOW-UP OF EARLY AND LONG-TERM TREATMENT WITH CAPTOPRIL ON THE PATIENTS WITH ACUTE MYOCARDIAL INFARCTION

Objective To investigate clinical outcomes of early and long-term treatment with captopril on patients with acute myocardial infarction (AMI) during a five-year follow-up. Methods In a randomi- zed trial, 822 patients (623 males, 199 females) with a first AMI with less 72h of symptoms were treated with captopril (treatment group, n=478, dosage from a first 6.25mg to 25mg t.i.d ) and conventional treatment (control group, n=344). Multivariable Cox’ regression were used to analyze relative risk of independent variables. Cumulative survival of both groups were calculated with Kaplan-Meier analysis and analyzed by using log-rank comparison. Results During the five-year follow-up, the age, Killip class (≥Ⅱ), anterior infarction, diabetes mellitus, and peak CPK increased relative risk of death after AML, but the effects of captopril, beta-blocker, antiplatelet drug, and thrombolytic therapy on the relative risk of death were contrary. The cumulative survival in different time during follow-up was higher in patients with captopril than controls (P< 0.001). Conclusion Early and long-term treatment with captopril was related to a beneficial outcome during the five-year follow-up after AMI.

Relationship between age and effect of early and long-term captopril treatment in patients with acute myocardial infarction

Abstract:Objective To analyse the relationship between age and treatment with captopril after acute myocardial infarction (AMI).Methods In a randomized trial, 822 patients with a first AMI received conventional medical treatment, including intravenous thrombolytic therapy and oral aspirin or metoprolol, and then were randomly allocated to captopril [dosage from the first 6.25?mg to 25?mg/t.i.d, 209 younger patients (≤64 years), 269 elderly patients (65-75 years)] or conventional treatment only (131 younger patients, 213 elderly). Survival in the four groups was calculated with the Kaplan-Meier method. Multivariate analysis was performed to understand the degree that multi-variables (including age) affect survival in patients taking captopril in the hospital or during long term follow-up.Results The survival of patients who took captopril correlated significantly with age (P<0.001). The survival of the elderly patients on captopril treatment did increase (P<0.0001), but not of the younger ones (P>0.05) during hospitalization. During follow-up, the survival of patients who took captopril correlated insignificantly with age (P>0.05), but both the elderly and the younger patients have good survival rates (all P<0.01) and lower cardiac events (all P<0.01) when they took captopril.Conclusions Captopril exerts a weak effect on the younger patients but a beneficial effect on the elderly patients during hospitalization after AMI. However, there is no difference between the younger and the elderly in the prognosis, both having good survival and lower cardiac events when they take captopril long term during follow-up.

ACE2/Ang-(1-7) signaling and vascular remodeling

The renin-angiotensin system(RAS)regulates vascular tone and plays a critical role in vascular remodeling,which is the result of a complex interplay of alterations in vascular tone and structure.Inhibition of the RAS has led to important pharmacological tools to prevent and treat vascular diseases such as hypertension,diabetic vasculopathy and atherosclerosis.Angiotensin converting enzyme 2(ACE2)was recently identified as a multifunctional monocarboxypeptidase responsible for the conversion of angiotensin(Ang)II to Ang-(1-7).The ACE2/Ang-(1-7)signaling has been shown to prevent cellular proliferation,pathological hypertrophy,oxidative stress and vascular fibrosis.Thus,the ACE2/Ang-(1-7)signaling is deemed to be beneficial to the cardiovascular system as a negative regulator of the RAS.The addition of the ACE2/Ang-(1-7)signaling to the complexities of the RAS may lead to the development of novel therapeutics for the treatment of hypertension and other vascular diseases.The present review considers recent findings regarding the ACE2/Ang-(1-7)signaling and focuses on its regulatory roles in processes related to proliferation,inflammation,vascular fibrosis and remodeling,providing proof of principle for the potential use of ACE2 as a novel therapy for vascular disorders related to vascular remodeling.

Correlativity between blood measures related to blood stasis blocking collaterals and gene expression of angiotensin-converting enzyme of renal cortex in diabetic rats and effect of stasis removing and collaterals dredging

OBJECTIVE: To investigate the correlativity between the changes of blood measures related to blood stasis blocking collaterals and gene expression levels of angiotensin-converting enzyme(ACE)and ACE2 ofrenal cortex in diabetic rats with stasis blocking collaterals syndrome, as well as the effectof stasis removing and collaterals dredging.METHODS: Male Sprague-Dawley rats were divided randomly into normal control group(C group),high-carbohydrate-fat control group(H group) and streptozotocin(STZ)-injecting group. The diabeticrats were induced from rats in the STZ-injecting groupby high-carbohydrate-fat diet combined with STZ intraperitoneal injection, with sustained high-carbohydrate-fat diet fed afterwards, and were further divided into model group(M group)and Chinese medicine of stasis removing and collaterals dredging group(Z group). Rats in the Z group were fed with stasis-removing-and-collaterals-dredging herbal granule suspension intragastrically daily for 16 weeks, while drinking water of corresponding volume was administrated to the rats in other groups. At the end of the 16 th week after successful establishment of models, the ultrastructures of glomeruli in different groups were detected by a transmission electron microscopy; and blood measures related to blood stasis blocking collaterals, including lipid profile and blood viscosity measures, were tested, as well as the relative gene expressions of ACE and ACE2.RESULTS: Changes in ultrastructures of glomeruli in the M group were characterized by lack of clarity in structure and occasional thickening of glomerular basement membrane and extensive fusion in foot processes. The correlation analysis showed that there were positive correlations between lipid profile, blood viscosity, and the ACE mRNA expression levels in the M group(P<0.05), except for cholesterol. And except for triglyceride, the blood measures were in negative correlation with the ACE2 mRNA expression levels in the M group(P<0.05).Compared with the C and H groups, the lipid profile, plasma viscosity and blood viscosity were significantly higher(P<0.01). All the above-mentioned measures were significantly improved in the Z group rats(P<0.05). ACE mRNA expression was significantly higher in the M group thanin the C group(P<0.05). ACE2 mRNA level was significantly lower in the M group than in the C and H groups(P<0.01)and its levelin the Z group was higher than that in the M group(P<0.01).CONCLUSION: Blood measuresrelated to blood stasis blocking collaterals had positive linear correlations with ACE mRNAexpression and negative linear correlations with ACE2 mRNA expression in the M group. Chinese recipe of stasis removing and collaterals dredging could play a renal protecting role for diabetic rats by reducing lipid profile and blood viscosity, down-regulating ACE mRNA expression and up-regulating ACE2 mRNA expression.