DETECTION OF RENIN mRNA IN ALDOSTERONE-PRODUCING ADENOMAS BY POLYMERASE CHAIN REACTION

1. Ribonucleic acid (RNA) was extracted from two normal human adrenal cortices and from five aldosterone-producing adenomas (APA). 2. The five APA could be categorized, on the basis of in vivo aldosterone responsiveness to angiotensin infusion and upright posture, into two APA responsive to both stimuli, two responsive only to angiotensin infusion, and one unresponsive to either stimulus. 3. RNA was reverse transcribed and coamplified by polymerase chain reaction (PCR) with an internal standard of renin complementary DNA (cDNA) containing a 60 base pair insertion. Renin mRNA in the APA was compared with normal adrenals. 4. Renin mRNA was greater than normal in the two APA responsive to both stimuli and less than, or similar to normal, in the two APA responsive only to angiotensin infusion. Renin mRNA was also less than, or similar to normal, in the APA unresponsive to either stimulus. 5. These findings support a possible role for adrenal renin in the development and biochemical behaviour of angiotensin-responsive APA.     

CORTISOL PRODUCTION BY ALDOSTERONE-PRODUCING ADENOMAS IN VITRO

1. In vitro short-term production of cortisol by dispersed tumour and non-tumorous adrenal cortical cells was measured with and without added angiotensin II (AII) or adrenocorticotrophin (ACTH) in adrenals removed from five patients with primary aldosteronism. 2. Aldosterone-producing adenomas (APA) were classified as angiotensin responsive (AII-R) or angiotensin unresponsive (AII-U) based on pre-operative behaviour in vivo. 3. Cortisol was produced by both tumour and cortex in vitro without stimulation, and significantly more cortisol was generated by the cortex. 4. Addition of AII significantly increased cortisol production by both tumour and cortex to an equal extent. 5. Addition of ACTH also significantly increased cortisol production by both tumour and cortex, but tumours were more responsive than cortex. The response to ACTH exceeded the response to angiotensin in both tumour and cortex. 6. There was no obvious difference between AII-R and AII-U APA in terms of cortisol production.     

Effects of Matrix Metalloproteinase (MMP)-2 Polymorphisms on Responsiveness to Antihypertensive Therapy of Women with Hypertensive Disorders of Pregnancy

Imbalanced matrix metalloproteinase (MMP) expression, including MMP‐2, has been demonstrated in pre‐eclampsia. However, little is known about the effect of polymorphisms in MMP‐2 gene on hypertensive disorders of pregnancy. We examined whether two functional MMP‐2 polymorphisms (g.‐1306C>T and g.‐735C>T) are associated with pre‐eclampsia and/or gestational hypertension and whether these polymorphisms affect therapeutic responses in women with these conditions. We studied 216 healthy pregnant women (HP), 185 patients with gestational hypertension (GH) and 216 patients with pre‐eclampsia (PE). They were stratified as responsive or non‐responsive to antihypertensive therapy according to clinical and laboratorial parameters of therapeutic responsiveness. Genomic DNA was extracted from whole blood and genotypes for g‐1306C>T and g.‐735C>T polymorphisms were determined by real‐time PCR using Taqman allele discrimination assays. Haplotype frequencies were inferred using the PHASE 2.1 program. The distributions of MMP‐2 genotypes and haplotypes were similar in HP, GH and PE patients (p > 0.05). In addition, we found no significant differences in MMP‐2 genotype or haplotype frequencies when GH or PE patients were classified as responsive or non‐responsive to antihypertensive therapy (p > 0.05). Our results suggest that MMP‐2 polymorphisms do not affect the susceptibility to hypertensive disorders of pregnancy. In parallel, MMP‐2 polymorphisms apparently do not affect the responsiveness to antihypertensive therapy of women with these hypertensive disorders of pregnancy.     

FAMILIAL HYPERALDOSTERONISM TYPE II: FIVE FAMILIES WITH A NEW VARIETY OF PRIMARY ALDOSTERONISM

1. Thirteen patients from five families had Familial Hyperaldosteronism Type II (FH-II), a new variety of familial primary aldosteronism not suppressible with dexamethasone that often involves adrenocortical adenoma formation. 2. Five patients had solitary aldosterone-producing adenomas, three had bilateral autonomous overproduction of aldosterone, and in five the subtype is yet to be determined. 3. Comparing FH-II patients with 88 patients with primary aldosteronism of other causes revealed no differences in mean age at presentation or at onset of hypertension, sex incidence, lowest recorded serum potassium, plasma aldosterone, plasma renin activity or adenoma size. 4. Analysis of DNA in peripheral blood of patients with FH-II, their affected and unaffected relatives, and in removed tumours is in progress in order to determine the underlying genetic defect(s) in FH-II, perhaps an abnormality in the P-450_aldo gene (CYP11B2). 5. It is recommended that hypertensive relatives of patients with primary aldosteronism should have measurements of the aldosterone/renin ratio.     

RENIN AND ANGIOTENSIN-CONVERTING ENZYME GENOTYPES IN PATIENTS WITH ESSENTIAL HYPERTENSION AND LEFT VENTRICULAR HYPERTROPHY

1. The associations between left ventricular hypertrophy (LVH) and specific alleles of the renin and angiotensin-converting enzyme (ACE) genes were studied in patients with essential hypertension and normal blood pressure. 2. LVH was present in 42% of those with essential hypertension (n= 72) and 17% of those with normal blood pressure (n= 44). 3. The frequency of each renin allele was the same in hypertensive and in normotensive patients. Renin allele frequencies were also the same for those with LVH and those with normal cardiac mass. When only hypertensives were considered, renin alleles were in the same proportion for the groups with and without LVH. Similarly, ACE alleles were not associated with essential hypertension nor with elevated cardiac mass. 4. We conclude that, in this population, variations in the renin or ACE genes do not contribute significantly to the development of LVH or to essential hypertension.     

ALDOSTERONE-PRODUCING ADENOMAS RESPONSIVE TO ANGIOTENSIN POSE PROBLEMS IN DIAGNOSIS

1. A subgroup of patients with aldosterone-producing adenoma (APA) have been identified who lack many of the biochemical features regarded as characteristic of APA and used to distinguish APA from bilateral adrenal hyperplasia. 2. In these patients, aldosterone is responsive to infused angiotensin II (angiotensin-responsive APA), which explains their uncharacteristic responses to upright posture, saline infusion and fludrocortisone acetate administration. 3. The angiotensin-responsiveness of these patients may derive from the contralateral adrenal gland, since renin levels are less completely suppressed in angiotensin-responsive APA than in angiotensin-unresponsive APA. 4. However, while the excretion of 18-oxo-cortisol was consistently increased in angiotensin-unresponsive APA, it was normal in angiotensin-responsive APA, consistent with biochemical and biosynthetic distinctiveness residing in the tumours. 5. Angiotensin-responsive APA should always be considered as an alternative diagnosis to bilateral hyperplasia causing primary aldosteronism.     

Intravitreal bevacizumab and dexamethasone implant for treatment of chronic diabetic macular edema

Objective: To evaluate anatomical and functional outcomes of intraviteal bevacizumab (IVB) in patients with chronic diabetic macular edema (DME), and the effectivity and safety of dexamethasone implant in those unresponsive to regular IVB treatment. Methods: Thirty-five eyes of 35 patients (16 male and 19 female) with chronic DME (central foveal thickness (CFT) >?275?μm, duration >?6 months) received three injections of 2.5?mg IVB with six-week intervals. At 18 weeks, dexamethasone implant was applied to patients unresponsive to IVB. Main outcomes were the change in best corrected visual acuity (BCVA), CFT and ocular and systemic adverse effects for both drugs. The patients responsive to IVB were followed up for 36 weeks and those patients receiving dexamethasone implant were followed up for 24 weeks postoperatively. Results: At 18 weeks, the mean BCVA (0.68?±?0.40 logMAR, p?=?0.45) and CFT (453?±?169?μm, p?=?0.58) did not show any significant change compared to baseline (0.74?±?0.42 logMAR and 521?±?151?μm, respectively). In 20 patients (%57.1) responsive to IVB, the CFT was significantly improved from 12 to 36 weeks with the mean value of 295?±?42 μ (p?=?0.01). However, no significant difference was observed for BCVA during this period (p?=?0.17). Dexamethasone was implanted in 15 eyes (42.8%) unresponsive to IVB at 18 weeks. Statistically significant improvements were observed in BCVA (at postoperative 4 and 12 weeks) and CFT (at postoperative 4, 12 and 24 weeks). In addition, both parameters significantly worsened at 24 weeks compared to 12 weeks (p?<?0.001 and p?=?0.01, respectively). Conclusions: Patients with chronic DME should be followed in accordance with a fixed treatment protocol combining anti-VEGF and steroid treatments.     

A RENIN-SECRETING TUMOUR SENSITIVE TO CHANGES IN CENTRAL BLOOD VOLUME (PRESUMABLY VIA SYMPATHETICS) BUT NOT TO CIRCULATING ANGIOTENSIN II

1. A 17 year old female presented with severe hypertension, hypokalaemia and elevated levels of plasma renin activity due to a renin-secreting tumour. 2. Renin was responsive to posture, low sodium diet, saline infusion and frusemide, but relatively unresponsive to raising or lowering circulating levels of angiotensin II. 3. Renal venous renin levels lateralized to the side of the tumour with good contralateral suppression when measured with control of posture and avoidance of prior stimulation, with and without angiotensin converting enzyme inhibition. 4. Levels of atrial natriuretic peptide were elevated and responsive to posture, saline infusion and angiotensin infusion. 5. The tumour was evident on computerized tomography, but not on intravenous pyelography or renal angiography. 6. Responsiveness of renin secretion to normal stimuli in reninoma may make diagnosis difficult, and renal vein sampling under controlled conditions is necessary.     

HAEMODYNAMIC, RENAL AND HORMONAL RESPONSES TO ENALKIREN IN FOUR PATIENTS WITH POST-SURGICAL OLIGURIA

1. The haemodynamic and hormonal responses of four patients with acute post-surgical oliguria (urine output <0.5 mL/kg perh) were measured in response to the renin inhibitor enalkiren. Enalkiren was infused at 0.01 up to 0.1 mg/kg perh for up to 4h. 2. Enalkiren infusion was associated with a progressive fall in blood pressure, clinically significant in three of the four patients. Systemic vascular resistance fell in proportion to blood pressure fall. Cardiac output and pulse rate remained unchanged. Effective renal plasma flow rose in all four cases (236 ± 19 to 327± 38). There was no change in urine flow rate, or urinary sodium excretion. 3. Plasma renin activity (ng angiotensin I/mL perh) fell from 1.9 ± 0.5 to 0.02 ± 0.01 (P<0.04), plasma angiotensin II (pg/mL) fell from 104 ± 93 to 7.7 ± 1.5, and plasma aldosterone (ng/dL) fell from 32 ± 8 to 21 ± 9 (P= 0.03) at the highest infusion dose. 4. Enalkiren inhibited plasma renin activity with reduced plasma angiotensin II and aldosterone concentrations. This was associated with vasodilation, reduced blood pressure and maintained cardiac output. There was no beneficial effect on renal function in these patients with post-surgical oliguria.     

DILATED CARDIOMYOPATHY WITH HYPERTENSION: PREVALENCE AND RESPONSE TO HIGH-DOSE β1-ADRENOCEPTOR ANTAGONIST THERAPY

• 1 The aim of the present study was to investigate the prevalence of hypertension in patients with dilated cardiomyopathy (DCM) and to determine the tolerance and efficacy of a high dose of the β₁‐adrenoceptor antagonist metoprolol in the long‐term treatment of DCM patients.
• 2 The prevalence of hypertension in DCM patients (n = 362) and age‐matched controls (n = 401) was evaluated and compared. To investigate the effects of metoprolol, DCM patients were divided into hypertensive (DCM‐H) or normotensive (DCM‐N) subgroups. Metoprolol was administered at a starting dosage of 6.25 mg/day and increased gradually to 250 mg/day or the maximum tolerable dose. Blood pressure (BP), heart rate (HR), left ventricular (LV) end‐diastolic dimension (LVEDD), left atrial end‐diastolic dimension (LAEDD), LV ejection fraction (LVEF), LV posterior wall thickness (LVPWT) and ventricular septal thickness (VST) were determined at baseline and 6 and 12 months after metoprolol treatment.
• 3 The prevalence of hypertension was significantly higher in DCM patients than in age‐matched controls (32.8 vs 20.1%, respectively; P < 0.01). Resting HR and a family history of hypertension were highest in the DCM‐H group. There were no significant differences in age, gender and occupation between the DCM‐H, DCM‐N and age‐matched control groups.
• 4 The tolerable dose for metoprolol was significantly higher in the DCM‐H group than the DCM‐N group (189.6 ± 14.8 vs 133.9 ± 12.0 mg/day, respectively; P < 0.05). Metoprolol significantly reduced BP and HR in the DCM‐H group and improved LVEDD, LAEDD and LVEF in all DCM patients, with a greater effect seen in the DCM‐H group.
• 5 In conclusion, DCM patients have a higher prevalence of hypertension than the general population. Patients in the DCM‐H subgroup were characterized by a higher resting HR and a family history of hypertension and were more tolerant of and more responsive to metoprolol treatment. These data suggest that this subgroup of DCM patients could have higher sympathetic nerve activity and is suitable for treatment with a higher dose of metoprolol.

     

ACUTE CHANGES IN BLOOD PRESSURE AND VASOACTIVE HORMONES AFTER CAPTOPRIL IN HYPERTENSIVE PATIENTS

1. The acute effects of captopril on blood pressure, renin, angiotensin, bra-dykinin and catecholamines were examined in patients with essential (n= 10, Group 1), accelerated (n=6, Group 2) and renal hypertension (n= 14, Group 3). 2. Blood pressure showed little change in Group 1, fell significantly in Group 2, with a marked fall in Group 3. Heart rate did not change. 3. The fall in blood pressure was positively correlated with pretreatment renin and angiotensin II. 4. Angiotensin II fell, with reciprocal increases in renin and angiotensin I. No changes occurred in bradykinin or catecholamines. 5. The rise in renin after captopril was greatest in renovascular hypertension which may prove useful as a screening test for such patients.     

Gene–Gene Interactions Among PRKCA,NOS3 and BDKRB2 Polymorphisms Affect the Antihypertensive Effects of Enalapril

Protein kinase C (PKC) signalling is critically involved in the control of blood pressure. Angiotensin‐converting enzyme inhibitors (ACEi) affect PKC expression and activity, which are partially associated with the responses to ACEi. We examined whether PRKCA (protein kinase C, alpha) polymorphisms (rs887797 C>T, rs1010544 T>C and rs16960228 G>A), or haplotypes, and gene–gene interactions within the ACEi pathway affect the antihypertensive responses in 104 hypertensive patients treated with enalapril as monotherapy. Patients were classified as poor responders (PR) or good responders (GR) to enalapril if their changes in mean arterial pressure were lower or higher than the median value, respectively. Multi‐factor dimensionality reduction was used to characterize interactions among PRKCA, NOS3 (nitric oxide synthase 3) and BDKRB2 (bradykinin receptor B2) polymorphisms. The TC+CC genotypes for the rs1010544 polymorphism were more frequent in GR than in PR (p = 0.037). Conversely, the GA+AA genotypes for the rs16960228 polymorphism, and the CTA haplotype, were more frequent in PR than in GR (p = 0.040 and p = 0.008, respectively). Moreover, the GG genotype for the PRKCA rs16960228 polymorphism was associated with PR or GR depending on the genotypes for the rs2070744 (NOS3) and rs1799722 (BDKRB2) polymorphisms (p = 0.012). Our results suggest that PRKCA polymorphisms and gene–gene interactions within the ACEi pathway affect the antihypertensive responses to enalapril.     

EFFECTS OF COMBINATION THERAPY WITH PERINDOPRIL AND LOSARTAN ON LEFT VENTRICULAR REMODELLING IN PATIENTS WITH MYOCARDIAL INFARCTION

• 1 Inhibiting the renin–angiotensin–aldosterone system prevents left ventricular (LV) remodelling after myocardial infarction (MI).
• 2 The present study was designed to assess the effects of a combination of perindopril and losartan on LV remodelling, cardiac function and serum procollagen type III amino terminal peptide (PIIINP) levels in patients with acute MI.
• 3 Patients with anterior MI were divided into three groups: (i) MI + perindopril; (ii) MI + losartan; and (iii) MI + perindopril + losartan. After successful intervention therapy, perindopril (2–4 mg daily), losartan potassium (25–50 mg daily) or their combination were administered. All patients received aspirin, clopidogrel and statins, and some patients were given beta‐blockers, nitrate and a platelet glycoprotein IIb/IIIa receptor antagonist. Three months later, LV dimensions and LV ejection fraction (LVEF) were measured by ultrasonography. Plasma B‐type natriuretic peptide (BNP), serum C‐reactive protein (CRP) and PIIINP levels were evaluated using enzyme‐linked immunosorbent assay or radioimmunoassay.
• 4 The baseline characteristics of the three groups were the same. Three months after the initiation of therapy, all patients showed decreased CRP, increased BNP and PIIINP levels and LV dilation and dysfunction. Compared with the two monotherapy groups, patients in the combination group showed significantly lower CRP, BNP and PIIINP levels, less LV dilation and higher LVEF. Serum PIIINP levels were positively correlated with CRP levels (r = 0.597; P < 0.01) and LV end‐diastolic volume index (r = 0.543; P < 0.01) and were negatively correlated with LVEF (r = –0.565; P < 0.01).
• 5 For patients with acute MI, combination treatment with perindopril and losartan significantly inhibited LV remodelling and improved LV function. Inhibition of myocardial interstitial fibrosis may be part of the underlying mechanism.

     

The association study of three FYN polymorphisms with prophylactic lithium response in bipolar patients

FYN belongs to the protein kinase family that phosphorylates NMDA receptor subunits, participating in the regulation of ion transmission and BDNF/TrkB signal transduction pathway. Lithium inhibits glutamatergic transmission via NMDA receptors, exerting neuroprotective effect against excitotoxicity. The aim of this study was to find possible association of three polymorphisms of FYN gene with prophylactic lithium response in the group of bipolar patients. We analyzed 101 bipolar patients treated with lithium carbonate for 5–27 years (mean 15 years). Twenty‐four patients were identified as excellent lithium responders (ER), 51 patients as partial responders (PRs), and 26 patients were non‐responders. Genotypes of the three analyzed polymorphisms were established by PCR‐RFLP. Statistical analysis was done with Statistica. No significant differences in genotype distribution and allele frequencies were observed between T/G and A/G FYN polymorphisms and lithium response. We observed a trend toward association of TT genotype and T allele of T/C polymorphism with worse lithium response. The results of the study demonstrated only marginal association between FYN polymorphisms and prophylactic lithium response in bipolar patients. The results are discussed in light of our previous studies on FYN gene in bipolar illness and BDNF gene in lithium response. Copyright © 2009 John Wiley & Sons, Ltd.     

NUDT15c.415C>T和TPMT*3C基因多态性检测方法的比较与临床应用

目的 建立准确、快速、经济的方法,检测NUDT15 c.415C>T和TPMT*3C基因多态性,探讨临床应用价值。方法 收集2017年5月-2018年5月期间福建汉族患者服用硫唑嘌呤2周以上的血清样本,提取DNA或白细胞后分别采用PCR-RFLP法、PCR-Sanger测序法和荧光定量PCR法对NUDT15 c.415C>T和TPMT*3C进行基因多态性分型,比较这3种方法的准确性、简便性及经济性。根据白细胞值分组,结合临床资料,探讨基因多态性等因素与硫唑嘌呤致白细胞减少的相关性。结果 共纳入129例患者,其中硫唑嘌呤致白细胞减少15例（11.6%）。3种方法的基因多态性检测结果一致,TPMT*3C未发现突变纯合子。携带NUDT15c.415C>T突变等位基因者服用硫唑嘌呤致白细胞减少的风险高于携带野生等位基因者（OR=6.2,95%CI：2.5~15.4,P=0.000 054）,而携带TPMT*3C突变等位基因者与野生等位基因者出现白细胞减少比例并无显著性差异（P=0.393）。NUDT15c.415C>T基因多态性预测白细胞减少敏感度为53.3%,特异度为85.1%,ROC曲线AUC为0.69。结论 3种方法都可用于临床检测NUDT15 c.415C>T和TPMT*3C基因多态性。PCR-RFLP法不需要专用试剂盒,也不需要昂贵的仪器设备,成本较低,过程简单,易于操作,特别适合条件有限的单位开展工作。福建汉族患者在服用硫唑嘌呤前进行NUDT15c.415C>T基因多态性检测比TPMT*3C更具临床价值。     

Direct renin inhibition: an evaluation of the safety and tolerability of aliskiren

ABSTRACT

Background: Aliskiren, an antihypertensive drug approved in the United States and Europe, is the first in a new class known as direct renin inhibitors. Aliskiren has been evaluated for safety and tolerability in more than 6400 patients. It has demonstrated a favorable safety and tolerability profile alone or in combination with other drugs.

Objective: This article reviews the currently available safety and tolerability data for aliskiren.

Methodology: Using the search term aliskiren, MEDLINE (no timeframe set) and major cardiovascular congresses (2005–2008) were searched. Articles and abstracts with safety and drug interaction data were included.

Findings: Aliskiren may share common adverse effects observed with angiotensin-converting enzyme (ACE)-inhibitor and angiotensin receptor blocker (ARB) therapy. In placebo-controlled trials, those commonly reported for aliskiren at the approved dosage were headache, diarrhea, and fatigue, with incidences similar to those of placebo. Aliskiren has been well tolerated in black, geriatric, diabetic, or obese patients and patients with renal or hepatic impairment. Aliskiren neither inhibits nor induces the cytochrome P450 system; it does not inhibit P-glycoprotein, but is a substrate for this drug transporter. Adding a direct renin inhibitor to another renin–angiotensin–aldosterone system (RAAS) inhibitor may further improve cardiovascular outcomes, renal outcomes, or both, without increasing the incidence of adverse effects.

Conclusions: Aliskiren is well tolerated, has an adverse effect profile comparable to that of placebo, and has a low potential for drug interactions. Data from ongoing trials evaluating the effects of aliskiren on surrogate markers, morbidity, and mortality will further define the role of direct renin inhibition in the antihypertensive armamentarium.     

Fine Mapping of Calcineurin (PPP3CA) Gene Reveals Novel Alternative Splicing Patterns,Association of 5′UTR Trinucleotide Repeat With Addiction Vulnerability,and Differential Isoform Expression in Alzheimer's Disease

Fine mapping of calcineurin (PPP3CA) gene identified single nucleotide polymorphisms (SNPs) and simple sequence repeat polymorphisms that are associated with addiction vulnerability. A trinucleotide repeat marker, located in the 5′untranslated region (5′UTR) of the PPP3CA mRNA, exhibited significantly different genotype and allele frequencies between abusers and controls in the NIDA African–American sample. The polymorphism showed allelic-specific expression in mRNA extracted from postmortem brain specimens. Novel alternatively spliced isoforms of PPP3CA were identified and their expressions were found altered in brain regions of postmortem Alzheimer's disease patients. These data underscore the importance of calcineurin gene in the molecular mechanism of addiction and Alzheimer's diseases.     

SLCO1B1基因多态性与阿托伐他汀的安全性及有效性相关性分析

目的 研究高脂血症患者SLCO1B1基因多态性与阿托伐他汀安全性及有效性的相关性。方法 收集金华市人民医院2017年4月—2018年4月在门诊确诊为高脂血症患者的基本资料,测定纳入患者的SLCO1B1 c.388A>G和c.521T>C的基因多态性,定期随访受试者,并定期测定其甘油三酯、胆固醇、低密度脂蛋白胆固醇及肌酸激酶等相关实验室检查指标。结果 纳入患者SLCO1B1 c.388A>G和c.521T>C等位基因频率分别为72.8%和15.9%。随访期结束后不同基因型患者的血清血脂指标变化率无明显差异。SLCO1B1 c.521T>C基因多态性与阿托伐他汀的安全性有相关性（P=0.005）。结论 SLCO1B1c.388A>G基因多态性对阿托伐他汀降脂疗效及安全性无影响。SLCO1B1 c.521T>C基因多态性与阿托伐他汀的降脂疗效无相关性,但对其安全性有一定影响。