Inhibition of glucose production and stimulation of bile flow by R (+)‐α‐lipoic acid enantiomer in rat liver

Abstract: Aims/Background: R (+)‐α‐lipoic acid (RLA) has been suggested for the treatment of liver diseases, but has also been shown to improve glucose utilization in diabetic patients. Because detailed information of RLA action on carbohydrate metabolism in intact liver is lacking, we examined concentration‐dependent effects of RLA on hepatic glucose production. Methods: RLA (10^?6?10^?3 mol L^?1) or buffer (control) was infused in isolated livers of fasted rats during recirculating perfusion for 90 min (n = 4–6/group). Hepatic glucose and lactate fluxes and bile secretion were continuously monitored. Results: RLA reduced lactate (10 mmol L^?1)‐dependent glucose production in concentration‐dependent fashion (R = ? 0.780, P < 0.001) by up to 67% compared with control (0.36 ± 0.02 µmol min^?1 g^?1). In parallel, RLA dose dependently decreased lactate uptake (R = ? 0.592, P < 0.001) also by up to 67% (control: 0.58 ± 0.08 µmol min^?1 g^?1). RLA (10^?4 mol L^?1 and 10^?3 mol L^?1) stimulated bile flow by ～ 20 and ～ 50%, respectively (P < 0.02 vs. control). After 10^?3 mol L^?1 RLA infusion, liver glycogen was ～ 3 fold higher (5.2 ± 1.1 vs. control: 1.8 ± 0.2 µmol g^?1, P < 0.002). Also at low lactate concentrations (1 mmol L^?1), 10^?3 mol L^?1 RLA reduced glucose production by ～ 53% and lactate uptake by ～ 60%, but stimulated bile secretion by ～ 50% (P < 0.05). Conclusion: RLA reduces hepatic glucose release by inhibiting lactate‐dependent glucose production in a concentration‐dependent fashion.     

Impact of non‐alcoholic fatty liver disease on microalbuminuria in patients with prediabetes and diabetes

Background: It is unknown whether microalbuminuria is associated with non‐alcoholic fatty liver disease (NAFLD) among patients with prediabetes and type 2 diabetes mellitus (DM). This study investigated the association of NAFLD with microalbuminuria among patients with prediabetes and diabetes. Methods: We evaluated 1361 subjects who had an abnormal oral glucose tolerance test (OGTT) on routine screening. All participants were divided into two groups, prediabetes and newly diagnosed type 2 DM, and the association of NAFLD with metabolic parameters on microalbuminuria was analysed. Results: The patients with NAFLD had higher prevalence rates of microalbuminuria (6.3% vs 19%; P= 0.001 in prediabetes, 4.5% vs 32.6%; P < 0.001 in diabetes) and also had a greater albumin‐to‐creatinine ratio (14.6 ± 52.0 µg/mg Cr vs 27.7 ± 63.9 µg/mg Cr; P= 0.051 in prediabetes, 11.4 ± 21.4 µg/mg Cr vs 44.7 ± 76.4 µg/mg Cr; P < 0.001 in diabetes) than those without NAFLD. The logistic regression analysis showed that NAFLD was associated with increased rates of microalbuminuria (odds ratio 3.66; 95% confidence interval (CI) 1.31–10.20, P= 0.013 in prediabetes, odds ratio 5.47; 95% CI 1.01–29.61, P= 0.048 in diabetes), independently of age, sex, body mass index, waist circumference, liver enzymes, lipid profiles, HbA1c, insulin resistance as estimated by homeostasis model assessment, hypertension, smoking status and the metabolic syndrome. Conclusions: The results of our study revealed a strong relationship between microalbuminuria and NAFLD in the patients with prediabetes and newly diagnosed diabetes. Further studies are required to confirm whether NAFLD is a predictor of the development of microalbuminuria in patients with prediabetes and diabetes.     

Effect of apoE/ATP‐containing liposomes on hepatic energy state

Background/Aims: ATP‐containing liposomes partially prevent ATP depletion in the cold‐stored liver. As hepatocytes can specifically bind apoE, we investigated whether the addition of apoE to large (200 nm) ATP‐containing liposomes increases their uptake by the liver and further improves hepatic energy stores. Methods: Livers from fasted male Hartley guinea‐pigs (231±3 g) were perfused for 90 min under our standard conditions (Control, n=6) or after a single bolus addition of plain liposomes (Lip, n=6), ATP (5 μmol)‐containing liposomes (ATP‐Lip, n=6) or apoE/ATP‐containing liposomes (0.8 or 8 mg apoE/g phospholipids; apoE1‐Lip and apoE10‐Lip, respectively, n=6 in each group). Liposome uptake and its impact on energy and nitrogen metabolism were studied. Results: At its highest concentration, apoE significantly increased liposome uptake (apoE10‐Lip:?9.17±0.69 vs apoE1‐Lip:?6.18±0.44 vs ATP‐Lip:?6.40±0.88 nmol min^?1 g^?1; P<0.05). This was associated with a significant increase in intrahepatic ATP (apoE10‐Lip: 1033±137 vs apoE1‐Lip: 811±98 and ATP‐Lip: 648±36 nmol g^?1; P<0.05), which was restored to its level in non‐perfused livers. Hepatic viability and nitrogen metabolism were not affected. Conclusions: Hepatic ATP content being a key factor in the maintenance of liver graft function, apoE/ATP‐containing liposomes should be useful in liver preservation for transplantation.     

Transcatheter aortic valve implantation for high risk patients With severe aortic stenosis using the Edwards Sapien balloon‐expandable bioprosthesis: A single centre study with immediate and medium‐term outcomes

Background : Transcatheter aortic valve implantation (TCAVI) is an emerging alternative therapy to open‐heart surgery in high‐risk patients with symptomatic aortic stenosis. Methods : Between January 2007 and May 2009, 46 patients underwent TCAVI with the 23 mm or 26 mm Edwards Sapien bioprosthesis via either the transapical (TA‐AVI) or transfemoral (TF‐AVI) approach. All patients had an estimated operative mortality risk of >15%. Results : A total of 46 patients (30 TA‐AVI, 16 TF‐AVI) with a mean aortic valve area (AVA) of 0.63 ± 0.2 cm² and mean gradient of 54 ± 16 mm Hg were treated. Predicted operative mortality was 25.3% by logistic Euroscore and 8.7% by Society of Thoracic Surgeons risk score. Procedural success was 93% in the TA‐AVI group and 88% in the TF‐AVI group. There was one intraprocedural death in the TA‐AVI group. Overall 30‐day mortality was 6.5% (2‐TA‐AVI, 1‐TF‐AVI). Four patients (9.5%) died from noncardiac causes after 30 days. Successful TCAVI was associated with a significant increase in AVA from 0.6 ± 0.1 cm² to 1.6 ± 0.6 cm² in the TA‐AVI group and 0.6 ± 0.1 cm² to 1.4 ± 0.2 cm² in the TF‐AVI group at a mean follow up of 7.4 ± 4.4 and 8.3 ± 5.0 months, respectively. At discharge, there was significant improvement in AVA (P < 0.0001), transaortic mean gradient (P < 0.0001), and mitral regurgitation (P = 0.01). At medium term follow up, the valve area was maintained and there was significant improvement in NYHA class in both groups (P < 0.0001). Conclusion : At medium term follow‐up, both transcatheter approaches demonstrated good valve durability with no cardiac‐related mortality post hospital discharge. © 2009 Wiley‐Liss, Inc.     

Measurement of spleen volume is useful for distinguishing between simple steatosis and early‐stage non‐alcoholic steatohepatitis

Aim: Although non‐alcoholic fatty liver disease (NAFLD) is now a common cause of chronic liver disease, discriminating between simple steatosis and non‐alcoholic steatohepatitis (NASH), especially early‐stage NASH, remains difficult. We investigated the clinical usefulness of measuring the spleen volume as a marker of early‐stage NASH. Methods: We evaluated computed tomography (CT) images obtained in 84 patients with histologically diagnosed NAFLD (22 with simple steatosis, 62 with NASH with mild fibrosis [stages 1–2]). We defined the data obtained by the following formula as a spleen‐body index (SBI): SBI = maximal CT axial section area of the spleen (cm²)/body surface area (BSA) (cm²) × 10⁴. We compared the SBI between patients with simple steatosis and those with NASH with mild fibrosis. Results: The mean SBI of the simple steatosis group was 15.8 ± 3.9, while that of the NASH with mild fibrosis group was 18.7 ± 5.7. This difference between the two groups was significant (P = 0.0314). A multiple logistic regression analysis showed that the SBI was significantly correlated with the discrimination of simple steatosis and NASH with mild fibrosis. The area under the receiver–operator curve was 0.661 for distinguishing between simple steatosis and NASH with mild fibrosis (P = 0.026, 95% confidence interval = 0.532–0.789). Conclusion: Spleen enlargement may be a distinct feature of NASH, especially early‐stage NASH. SBI might be a non‐invasive and simple method of differentiating NASH and simple steatosis.     

Adipokines in type 1 diabetes after successful pancreas transplantation: normal visfatin and retinol‐binding‐protein‐4, but increased total adiponectin fasting concentrations

Objective In type 1 diabetes mellitus (T1DM), the release of many hormones, not only from beta‐cells, but also from adipocytes (adipokines) may be altered. After successful pancreas–kidney‐transplantation (PKTx), T1DM patients can revert to a nondiabetic metabolism, but it is unclear whether alterations of adipokines are still present after PKTx. Design, patients and measurements Concentrations of adipokines [visfatin, retinol‐binding protein‐4 (RBP‐4), adiponectin, high molecular weight (HMW) adiponectin] were measured at fasting in 10 PKTx and in 19 T1DM. Nondiabetic healthy controls (CON, n = 9) and six nondiabetic patients after kidney transplantation (KTx) were examined as control groups. In PKTx, KTx and CON, indices of insulin sensitivity (OGIS) and beta cell function (adaptation index, AI) were calculated from 75 g oral glucose tolerance test (OGTT) data. Results Fasting serum visfatin (T1DM: 56 ± 4 μg/l, PKTx: 42 ± 6 μg/l, KTx: 39 ± 3 μg/l, CON: 40 ± 3 μg/l) and RBP‐4 (T1DM: 490 ± 26 μg/l, PKTx: 346 ± 39 μg/l, KTx: 401 ± 13 μg/l, CON: 359 ± 36 μg/l) was increased by 40% and 36%, respectively (each P < 0·03) in T1DM only. Levels were positively correlated with HbA1c in all subjects (visfatin: r = 0·43, P < 0·004; RBP‐4: r = 0·46, P < 0·03). Fasting plasma adiponectin was 80% higher in T1DM and in PKTx (T1DM: 18 ± 2 mg/l, PKTx: 18 ± 3 mg/l, KTx: 12 ± 3 mg/l, CON: 10 ± 1 mg/l; P < 0·04) and was positively correlated with diabetes duration (r = 0·37, P < 0·02). HMW/total adiponectin ratio was increased in T1DM (P < 0·02). PKTx displayed a normoglycaemic metabolism as insulin sensitive as CON, but AI was lower than in CON and KT (P < 0·01). Conclusions T1DM after successful PKTx show normal fasting visfatin and RBP‐4 levels and HMW‐adiponectin/adiponectin‐ratio, which are elevated in T1DM, whereas total adiponectin levels are similarly increased in T1DM and PKTx patients.     

Longitudinal evolution of vertically HIV/HCV–co‐infected vs HCV–mono‐infected children

HIV co‐infection has been suggested to play a deleterious role on the pathogenesis of liver fibrosis among vertically HCV‐infected children. The aim of this study was to describe the longitudinal evolution of vertically acquired HIV/HCV co‐infection in youths, in comparison with HCV infection alone. This was a retrospective, multicentre study including vertically HIV/HCV–co‐infected patients and age‐ and sex‐matched vertically HCV–mono‐infected patients. Progression to advanced liver fibrosis, defined as F3 or more by elastography or METAVIR biopsy staging, and response to treatment were compared by means of univariate and multivariate regression analyses and Cox regression models. Sixty‐seven co‐infected patients were compared with 67 matched HCV–mono‐infected patients. No progression to advanced liver disease was observed during the first decade. At a median age of 20.0 [19.0, 22.0] years, 26.7% co‐infected vs 20% mono‐infected had progressed to advanced fibrosis (P = .617). Peg‐IFN/RBV for HCV treatment was given to 37.9% vs 86.6% (P‐value < .001). At treatment initiation, co‐infected patients were older (16.9 ± 4.1 vs 11.7 ± 4.5 years, P < .001), and 47.1% vs 7.1% showed advanced fibrosis (P < .003), with no differences in hard‐to‐treat genotype distribution. Sustained viral response was comparable between groups (43.5% vs 44.0%, P = .122). In vertically HIV/HCV–co‐infected patients, the progression to liver fibrosis was rare during childhood. At the end of adolescence, over 25% of patients displayed advanced liver disease. Response to Peg‐IFN/RBV was poor and comparable in both groups, supporting the need for fast access to early treatment with direct‐acting antivirals against HCV for vertically co‐infected patients.     

Fibrosis evolution in chronic hepatitis B e antigen‐negative patients across a 10‐year interval

The degree of liver fibrosis in chronic hepatitis B (CHB) infection influences outcome and management. Existing data describing the long‐term dynamic changes of liver fibrosis are limited. This study aimed to evaluate the evolution of liver fibrosis in CHB across a 10‐year period. CHB patients with liver stiffness measurement (LSM) by transient elastography 10 years ago were recruited for follow‐up LSM. Fibrosis stages were classified according to EASL‐ALEH guidelines. Fibrosis progression/regression was arbitrarily defined as ≥1 fibrosis stage change from baseline. A total of 459 hepatitis B e antigen (HBeAg)‐negative patients (224 untreated, 235 treated with nucleos(t)ide analogues [NAs]) were recruited. The mean age at baseline LSM was 41.7 ± 9.0 years (56.2% male). Over 10 years, the proportion of patients with advanced fibrosis/cirrhosis significantly reduced from 16.3% to 5.7% (P < 0.001). Fibrosis progression and regression were observed in 8.7% and 37.5%, respectively. No treatment with NAs (OR 2.259, 95% confidence interval [CI]: 1.032‐4.945), metabolic syndrome (OR 4.379, 95% CI: 1.128‐16.999) and hepatic steatosis (OR 7.799, 95% CI: 2.271‐26.776) was associated with fibrosis progression. Liver stiffness decline demonstrated positive correlation with the time after HBsAg seroclearance (r = ?0.50, P < 0.001). Median liver stiffness was higher both at baseline (14.0 vs 6 kPa, P < 0.001) and 10 years (9.1 vs 4.9 kPa, P < 0.001) in patients with cirrhosis‐related complications/hepatocellular carcinoma compared with those without. In conclusion, CHB‐related liver fibrosis changed dynamically across 10 years. Metabolic syndrome and hepatic steatosis were associated with fibrosis progression, while antiviral therapy was associated with fibrosis regression. Patients with HBsAg seroclearance demonstrated time‐dependent decline in liver stiffness.     

Plasma soluble tumour necrosis factor‐α receptor 2 is elevated in obesity: specific contribution of visceral adiposity

Objective We examined the obesity phenotype most strongly associated with increased plasma concentrations of sTNFR2, and compared which of the two markers, TNF‐α or sTNFR2, better predicts indices of plasma glucose‐insulin homeostasis. Design, patients and measurements Plasma sTNFR2 levels were measured in a sample of 287 healthy nondiabetic men [age: 43·9 ± 8·0 years (mean ± SD)], covering a wide range of adiposity values (BMI: 29·0 ± 4·4 kg/m²; waist girth: 100·0 ± 11·7 cm). Results Plasma sTNFR2 levels correlated positively and significantly with BMI (r = 0·36; P < 0·0001), fat mass (r = 0·42; P < 0·0001), waist girth (r = 0·38; P < 0·0001) as well as with visceral (r = 0·37; P < 0·0001) and subcutaneous adipose tissue (AT) (r = 0·40; P < 0·0001) areas measured by computed tomography. Two subgroups (n = 27 in each group) of overweight men (BMI ≥25 kg/m²) were individually matched for similar BMI values, but with markedly different levels of visceral AT (< or ≥130 cm²) and then compared with a control group of 46 lean subjects (with both BMI <25 kg/m² and visceral AT <130 cm²). This analysis revealed that men characterized by high levels of visceral AT had significantly higher concentrations of sTNFR2 compared with obese men with low visceral AT (1861 ± 457 pg/ml vs. 1722 ± 400; P < 0·05) and with lean controls (1570 ± 291 pg/ml; P < 0·001). Whereas subjects classified across tertiles of TNF‐α levels showed no difference in glucose tolerance and insulin levels, subjects in the upper tertile of plasma sTNFR2 levels were characterized with the highest plasma insulin concentrations during the OGTT and had the highest area under the curve of insulin concentrations. Conclusions These results indicate that sTNFR2 levels are more closely related to abdominal AT accumulation than to total adiposity. Furthermore, plasma concentrations of sTNFR2 are independently related to plasma glucose‐insulin homeostasis beyond the known contribution of visceral adiposity.     

Role of angiotensin‐1 receptor blockade in cirrhotic liver resection

Background: The regeneration capacity of cirrhotic livers might be affected by angiotensin‐1 (AT1) receptors located on hepatic stellate cells (HSC). The effect of AT1 receptor blockade on microcirculation, fibrosis and liver regeneration was investigated. Materials and methods: In 112 Lewis rats, cirrhosis was induced by repetitive intraperitoneal injections of CCl₄. Six hours, 3, 7 and 14 days after partial hepatectomy or sham operation, rats were sacrificed for analysis. Animals were treated with either vehicle or 5 mg/kg body weight losartan pre‐operatively and once daily after surgery by gavage. Microcirculation and portal vein flow were investigated at 6 h. The degree of cirrhosis was assessed by Azan Heidenhein staining, activation of HSC by desmin staining, apoptosis by ssDNA detection and liver regeneration by Ki‐67 staining. Changes in expression of various genes important for liver regeneration and fibrosis were analysed at 6 h and 3 days. Haemodynamic parameters and liver enzymes were monitored. Results: Losartan treatment increased sinusoidal diameter, sinusoidal blood flow and portal vein flow after partial hepatectomy (P<0.05), but not after sham operation. AT1 receptor blockade resulted in increased apoptosis early after resection. HSC activation was reduced and after 7 days, a significantly lower degree of cirrhosis in resected animals was observed. Losartan increased the proliferation of hepatocytes at late time‐points and of non‐parenchymal cells early after partial hepatectomy (P<0.05). Tumour necrosis factor (TNF)‐α was significantly upregulated at 6 h and stem cell growth factor (SCF) was downregulated at 3 days (P<0.05). Conclusion: Losartan increased hepatic blood flow, reduced HSC activation and liver fibrosis, but interfered with hepatocyte proliferation after partial hepatectomy in cirrhotic livers.     

Immediate and 18‐Month Outcome of Balloon Mitral Valvuloplasty: Comparison of Inoue and Multi‐Track System

Aims: To compare the immediate and 18‐month clinical and echocardiographic outcome of Inoue and multi‐track system for balloon mitral valvuloplasty (BMV). Methods: We included 78 consecutive patients with moderate to severe rheumatic mitral stenosis (MS) [mitral valve area (MVA) < 1.5 cm²] and clinically indicated BMV. The first 42 consecutive patients were assigned to Inoue BMV (group I), and the following 36 consecutive patients were assigned to multi‐track system (group M). Clinical and echocardiographic assessment was performed before, immediately after, 3 months after, and 18 months after the procedure. Results: The successful immediate result [MVA > 1.5 cm² and mitral regurgitation (MR) < II/IV] was achieved in 40 (95.23%) patients of group I and 34 (94.44%) patients of group M (P = 0.12). Immediately after BMV, MVA increased from 0.9 ± 0.4 to 1.7 ± 0.5 cm² in group I and from 0.8 ± 0.2 to 1.9 ± 0.3 cm² in group M (P < 0.01). Bilateral commissural splitting was significantly higher in group M (P < 0.01). This was associated with higher incidence of mild commissural mitral regurgitation. There were no significant differences of moderate to severe MR. Both procedure and fluoroscopy time were significantly shorter in group I (P < 0.001). Eighteen‐month clinical and echocardiographic evaluation was available for 66 (84.64%) patients with sustained immediate clinical and echocardiographic improvements. Conclusions: Both Inoue and the multi‐track balloon systems achieved successful immediate and 18‐month results. The multi‐track double balloon system produced significantly larger MVA, with better bilateral commissurotomy, yet with longer procedure and fluoroscopy times. (J Interven Cardiol 2012;25:47–52)     

Expansion of CD4+CD25+FoxP3+ regulatory T cells in hepatitis C virus‐related chronic hepatitis,cirrhosis and hepatocellular carcinoma

Aim: Regulatory T (Treg) cells may play a pivotal role in the persistence of hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC). Therefore, we examined their frequency in peripheral blood from patients with HCV‐positive chronic hepatitis (CH), cirrhosis (LC) and HCC. Methods: Treg cells were identified as CD4⁺, CD25⁺ and FoxP3⁺ T lymphocytes using three‐color FACS. The frequency of Treg cells was expressed as a percentage of the total CD4⁺ T lymphocytes, and the phenotype of Treg cells was examined using CD45RA. Results: Treg cells were significantly increased in CH (5.88 ± 0.19%, n = 76; P < 0.01), LC (6.10 ± 0.28%, n = 40; P < 0.001) and HCC (6.80 ± 0.30%, n = 57; P < 0.0001) compared to healthy control (5.13 ± 0.25%, n = 31). However, Treg cells were not increased with the progression of fibrosis or the grade of inflammations. Treg cells were slightly increased in early‐stage HCC (6.91 ± 0.40%) compared with advanced‐stage HCC (6.58 ± 0.39%), but these results were not statistically significant. In a serial examination, a distinct increase in Treg cells after local therapy for early‐stage HCC was a hallmark of early recurrence. Most expanded Treg cells in HCC were CD45RA^‐, suggesting that a memory‐type Treg population had differentiated in the periphery and not in the thymus. Conclusion: We observed an increase in Treg cells in HCV‐related chronic liver disease, particularly in HCC, and these cells were shown to be memory‐type Treg cells.     

Night‐time sleep disturbance does not correlate with neuropsychiatric impairment in patients with cirrhosis

Background: Sleep–wake disturbances are common in patients with cirrhosis and are generally attributed to the presence of hepatic encephalopathy. Aim: To determine the relationship between sleep and neuropsychiatric disturbances in patients with cirrhosis. Methods: The study population comprised 87 patients, classified as neuropsychiatrically unimpaired or as having minimal/overt hepatic encephalopathy. Nineteen healthy volunteers served as controls. Validated questionnaires were used to assess sleep quality [Pittsburgh sleep quality index (PSQI)], day‐time sleepiness [Epworth sleepiness scale (ESS)] and diurnal preference. Health‐related quality of life (H‐RQoL) was assessed using the 36‐item short form health profile (SF‐36v1) and the chronic liver disease questionnaire. Results: Patients slept significantly less well than the healthy volunteers (PSQI score: 8.4 ± 4.9 vs. 4.6 ± 2.5, P<0.01) and had more pronounced day‐time sleepiness (abnormal ESS: 21 vs. 0%; χ²=3.8, P=0.05). No significant relationships were observed between sleep indices and the presence/degree of hepatic encephalopathy. H‐RQoL was significantly impaired in the patients (SF‐36v1 physical score: 36 ± 15 vs. 50 ± 10, P<0.001; SF‐36v1 mental score: 46 ± 11 vs. 50 ± 10, P<0.01); night‐time sleep disturbance was an independent predictor of poor H‐RQoL (P<0.01). Conclusions: Sleep–wake abnormalities are common in patients with cirrhosis; they significantly affect H‐RQoL but are not related to the presence of hepatic encephalopathy.     

Hepatoprotective effect of a curcumin/absinthium compound in experimental severe liver injury

OBJECTIVE: A preliminary in vitro study with hepatocyte culture showed that concentrations as low as 10 µg/mL of PN‐M001 are able to significantly mitigate CCl4 hepatocyte damage (P < 0.05) comparable to 100 µg/mL silymarin, and 100 µg/mL proved to be more protective than either silymarin 100 µg/mL or glycyrrhizin 10 µg/mL (P < 0.05). METHODS: Wistar rats were allocated into three groups: (A) 0.1 mL/100 g body weight (BW) mixture of CCl₄ in olive oil (1 : 1 v/v) subcutaneous injection twice daily for 4 weeks; (B) as A, plus oral administration of 50 mg/kg of K‐17.22 dissolved in 5% glucose; (C) as B but with PN‐M001 given 1 week after the first injection of CCl₄. Rats were killed at the end of the study and blood and liver samples were obtained. RESULTS: When compared with a control, group A showed a significant decrease of glutathione (GSH;>45%, P < 0.001) and oxidized GSH (GSSG; P < 0.01) liver content, a lower liver wet weight (P < 0.01) together with an increase of both transaminases (>15‐fold, P < 0.001) whereas groups B and C both showed only a mild increase in transaminases (<4‐fold, P < 0.05). Group A showed a significant decrease of Y‐protein fraction and of GST activity, as tested by both substrates (P < 0.01 vs control). However, both these parameters were reverted to normal by PN‐M001 (P < 0.05 vs A). CONCLUSIONS: These preliminary data suggest that PN‐M001 exerts a highly protective and prolonged effect (either preventive or therapeutic) on GSH depletion in CCl₄‐induced liver injury, which suggests its potential use in the clinical setting.     

Arterial duct stenting in low‐weight newborns with duct‐dependent pulmonary circulation

Objectives: To evaluate feasibility and results of arterial duct (AD) stenting in low‐weight newborns with congenital heart disease and duct‐dependent pulmonary circulation (CHD‐DPC). Background: AD stenting is nowadays considered a cost‐effective alternative to surgical shunt in CHD‐DPC. This option might be even more advisable in low‐weight neonates (<2.5 kg), who are at higher surgical risk and in whom stent redilation might adapt shunt magnitude to patient's growth. Methods: Between April 2003 and September 2010, 76 neonates with CHD‐DPC underwent AD stenting at our institution, as lower‐risk palliation with respect to surgical shunt. Procedural and follow‐up data of the 15 low‐weight newborns (2.0 ± 0.3 kg, median 2.2) (group I) were compared with the remaining normal‐weight newborns (3.5 ± 0.7 kg, median 3.2) (group II). Results: Feasibility, complication rate, and need for surgical shunt did not significantly differ between groups. Global X‐ray exposure was significantly higher in the low‐weight group (82 ± 108 vs. 30 ± 33 Gray/cm², P < 0.002), which maybe due to a longer angiographic presenting work‐up. In‐hospital mortality rate was 14.3% (vs. 1.9% in the group II, P = NS), although none of the fatalities was procedure‐related. During follow‐up, five patients (35.7% vs. 15.7% in the group II, P = NS) underwent stent redilation before surgical repair. At control angiography, the Nakata and McGoon indexes had significantly increased (P < 0.05 for both comparisons), without any significant difference with the group II (162 ± 52% vs. 144 ± 158% and 40 ± 17% vs. 42 ± 38%, P = NS). Conclusions: AD stenting is also feasible and effective in low‐weight newborns with CHD‐DPC, supporting the spontaneous improvement process or promoting a significant pulmonary artery growth. © 2011 Wiley‐Liss, Inc.     

Steatosis correlates with hepatic expression of death receptors and activation of nuclear factor‐κB in chronic hepatitis C

Background: Steatosis is recognized as a predictor of the severity as well as the progression of fibrosis in chronic hepatitis C. The mechanisms that cause increased hepatocellular injury associated with steatosis remain largely unknown. Methods: We studied the correlation of hepatic expression of death receptors: Fas and tumour necrosis factor‐α receptor 1 (TNF‐R1), and downstream caspase (caspase‐3) with hepatic steatosis by immunohistochemical study in chronic hepatitis C and determined the role of nuclear factor‐κB (NF‐κB). Results: Ninety patients (49 males and 41 females, mean age of 50.5 ± 10.4 years, genotype 1 or 2) with chronic hepatitis C virus infection were recruited. The factors associated with steatosis grade were body mass index (P=0.004) and fibrosis stage (P=0.034). Moderate/severe steatosis was an independent variable associated with advanced fibrosis stage by stepwise logistic regression analysis. The expression of immunoreactivity for Fas, TNF‐R1 and active caspases‐3 in liver tissues was significantly correlated with the steatosis grade (P<0.001, P<0.001 and P<0.001 respectively). The extent of active caspases‐3 correlated significantly with the expression of Fas (r=0.659, P<0.001) and TNF‐R1 (r=0.617, P<0.001). NF‐κB p65 expression correlated significantly with the extent of Fas (r=0.405, P<0.001), TNF‐R1 (r=0.448, P=0.002) and active caspase‐3 (r=0.313, P=0.003), and correlated with steatosis grade (P<0.001) but not with inflammatory and fibrosis scores. Conclusion: Our observations suggest a mechanism whereby steatosis contributes to the progression of liver injury in chronic hepatitis C through upregulation of death receptors and activation of NF‐κB.     

Serum Wisteria floribunda agglutinin‐positive Mac‐2‐binding protein for patients with chronic hepatitis B and C: a comparative study

We compared Wisteria floribunda agglutinin‐positive Mac‐2‐binding protein (WFA⁺‐M2BP) levels between patients with chronic hepatitis B (n=249) and chronic hepatitis C (n=386) based on the degree of liver fibrosis. We examined WFA⁺‐M2BP levels in patients with F4 (cirrhosis), F3 or more (advanced fibrosis) and F2 or more (significant fibrosis) in the two groups. We further examined the relationship between five fibrosis markers and the degree of fibrosis. The WFA⁺‐M2BP values ranged from 0.25 cut‐off index (COI) to 12.9 COI in patients with hepatitis B and 0.34–20.0 COI in patients with hepatitis C (P<.0001). The median WFA⁺‐M2BP values in F4 in the two groups were 2.83 COI in patients with hepatitis B and 5.03 COI in patients with hepatitis C (P=.0046). The median WFA⁺‐M2BP values in F3 or more in the two groups were 1.79 COI in patients with hepatitis B and 3.79 COI in patients with hepatitis C (P<.0001). The median WFA⁺‐M2BP values in F2 or more in the two groups were 1.49 COI in the hepatitis B cohort and 3.19 COI in the hepatitis C group (P<.0001). Among five liver fibrosis markers, WFA⁺‐M2BP had the highest correlation coefficient (r_s=.629) in terms of correlation with the degree of fibrosis in the patients with hepatitis C and had the second highest r_s value (.415) in the hepatitis B group. Although WFA⁺‐M2BP could be a useful indicator of liver fibrosis, WFA⁺‐M2BP levels in the two groups significantly differed even in the same degree of fibrosis. Individual cut‐off values in each aetiology for the degree of fibrosis should be determined.     

Long‐Term Effects of Renal Sympathetic Denervation on Hypertensive Patients With Mild to Moderate Chronic Kidney Disease

Thirty patients who underwent percutaneous renal denervation, which was performed by a single operator following the standard technique, were enrolled in this study. Patients with chronic kidney disease (CKD) stage 2 (n=19), 3 (n=6), and 4 (n=5) were included. Data were obtained at baseline and at monthly intervals for the first 6 months. At 7 months, follow‐up data were collected bimonthly until month 12, after which data were collected on a quarterly basis. Baseline blood pressure values (mean±standard deviation) were 185±18/107±13 mm Hg in the office and 152±17/93±11 mm Hg through 24‐hour ambulatory blood pressure monitoring (ABPM). Three patients with stage 4 CKD required chronic renal replacement therapy (one at the 13‐month follow‐up and two at the 14‐month follow‐up) after episodes of acute renal injury; their follow‐up was subsequently discontinued. The office blood pressure values at the 24‐month follow‐up were 131±15/87±9 mm Hg (P<.0001, for both comparisons); the corresponding ABPM values were 132±14/84±12 mm Hg (P<.0001, for both comparisons). The mean estimated glomerular filtration rate increased from 61.9±23.9 mL/min/1.73 m² to 88.0±39.8 mL/min/1.73 m² (P<.0001). The urine albumin:creatinine ratio decreased from 99.8 mg/g (interquartile range, 38.0–192.1) to 11.0 mg/g (interquartile range, 4.1–28.1; P<.0001 mg/g). At the end of the follow‐up period, 21 patients (70% of the initial sample) were no longer classified as having CKD.     

Effects and mechanism of the selective COX‐2 inhibitor,celecoxib, on rat colitis induced by trinitrobenzene sulfonic acid

OBJECTIVE: To investigate the action of celecoxib (a selective COX‐2 inhibitor) in a rat model of colitis induced by trinitrobenzene sulfonic acid (TNBS). METHODS: Rats were randomized into four groups. Colitis was induced in groups 1 and 2 by intracolonic administration of TNBS (25 mg/mL) in 50% ethanol (0.25 mL). The rats in group 1 received oral celecoxib (1.25 mg/kg) and those in group 2 received distilled water (1 mL/0.3 kg), beginning 3 h before induction of colitis and continuing twice daily thereafter for up to 7 days. The rats in group 4 received oral celecoxib (1.25 mg/kg) twice daily for 7 days and those in group 3 were healthy controls. All rats that survived 7 days were killed and both the severity of colonic mucosal damage and the prostaglandin E₂ (PGE₂) concentrations of the colonic mucosa were assessed. RESULTS: The colonic mucosal damage scores for groups 1 and 2 were 11.15 ± 3.30 and 8.50 ± 2.82, respectively, both of which were significantly higher than the score for the healthy controls (0.62 ± 0.09; P < 0.01, P < 0.01). The score of group 1 was significantly higher than that of group 2 (P < 0.05). No difference was found between the scores of groups 3 and 4. The mucosal concentrations of PGE₂ in groups 1 and 2 were 12.00 ± 4.33 pg/µg and 17.20 ± 9.62 pg/µg, respectively, both of which were significantly higher than the concentration in the healthy controls (6.02 ± 3.39 pg/µg; P < 0.05, both). The PGE₂ concentration of group 1 was decreased significantly compared with that of group 2 (P < 0.05). No difference was found between groups 3 and 4. CONCLUSION: The results suggest that treatment with celecoxib exacerbates inflammation‐associated colonic injury in experimental colitis induced by TNBS. This preliminary study shows that the mechanism is related to suppression by the COX‐2 inhibitor of the PG derived from COX‐2, but further study is needed to identify if there are other related mechanisms.     

Trichosanthin‐induced apoptosis in gastric cancer is related to the downexpression of bcl‐2

OBJECTIVE : To study: (i) the induction of apoptosis in gastric cancer cells by trichosanthin; and (ii) the relationship between apoptosis and the expression of bcl‐2. METHODS : During in vitro experiments, morphological studies and the terminal deoxynucleotidyl transferase‐mediated dUTP–digoxigenin nick end‐labeling (TUNEL) method were used to detect apoptosis in gastric adenocarcinoma cell line SGC‐7901 before and after trichosanthin treatment. An immunohistochemical staining method and northern blot hybridization were used to detect the expression of the apoptosis‐related gene bcl‐2 before and after trichosanthin treatment. RESULTS : When SGC‐7901 cells were treated with trichosanthin (0.1 μg/mL, 36 h), they presented some typical apoptotic morphological changes that were observed by fluorescent staining. These morphological changes included nuclear condensation and nucleosomal fragments forming a lunate body under the nuclear membrane. When SGC‐7901 cells were treated with trichosanthin (0.1 μg/mL) for 36, 42 or 48 h, TUNEL staining revealed a significant increase in the apoptotic index (AI), from 3.78 ± 1.11%, 3.98 ± 1.12% and 3.85 ± 1.08%, to 11.30 ± 2.33%, 10.22 ± 2.00% and 11.18 ± 1.85% (P < 0.01), respectively. When SGC‐7901 cells were treated with trichosanthin (0.1 μg/mL, 32 h), immunohistochemical staining revealed a decreased expression of the bcl‐2 protein product: the staining density decreased from ++/+++ to –/+ (P < 0.01). When SGC‐7901 cells were treated with trichosanthin (0.1 μg/mL, 24 h), northern blot hybridization showed a decreased expression of bcl‐2 RNA: hybridization decreased from 35.19 ± 2.34 to 22.27 ± 3.90 (P < 0.01). CONCLUSIONS : Trichosanthin is able to induce apoptosis in gastric cancer. The apoptosis may be mediated by the downexpression of the apoptosis‐related gene bcl‐2.