Effects of Selenium Supplementation on the Ion Homeostasis in the Reproductive Organs and Eggs of Laying Hens Fed With the Diet Contaminated With Cadmium, Lead, Mercury, and Chromium.

The objective of this study was to explore the toxic effects of different heavy metals in combination with their deposition and ion homeostasis in the reproductive organs and eggs of laying hens, as well as the alleviating action of selenized yeast. A total of 160 Lohmann pink-shell laying hens (63-week-old) were randomly allocated into four treatments with 10 replicates of four hens each. The four dietary treatments were the corn-soybean meal basal dietary (control; CON); the CON dietary supplemented with 0.4 mg/kg selenium from selenized yeast (Se); the CON dietary supplemented with 5 mg/kg Cd + 50 mg/kg Pb +3 mg/kg Hg + 5 mg/kg Cr (HEM), and the HEM dietary supplemented with 0.4 mg/kg selenium from selenized yeast (HEM+Se). The dietary HEM significantly increased Cd, Pb, and Hg deposition in the egg yolk and ovary, and Cd and Hg deposition in the oviduct and in the follicular wall (p < 0.05). The HEM elevated Fe concentration in the egg yolk, ovary, and oviduct (p < 0.05). The HEM decreased Mn concentration in the egg yolk, Fe, Mn, and Zn concentrations in the egg white, Cu concentration in the ovary, Mg concentration in the oviduct, as well as Ca, Cu, Zn, and Mg concentrations in the follicular walls (p < 0.05). Dietary Se addition elevated Se concentration in the egg yolk, oviduct, and follicular walls and Mg concentration (p < 0.05) in the oviduct, whereas it reduced Fe concentration in the oviduct compared with the HEM-treated hens. Some positive or negative correlations among these elements were observed. Canonical Correlation Analysis showed that the concentrations of Pb and Hg in the egg yolk were positively correlated with those in the ovary. The concentration of Cd in the egg white was positively correlated with that in the oviduct. In summary, dietary Cd, Pb, Hg, and Cr in combination caused ion loss and deposition of HEM in reproductive organs of laying hens. Dietary Se addition at 0.4 mg/kg from selenized yeast alleviated the negative effects of HEM on Fe and Mg ion disorder in the oviduct and follicle wall of hens.

Diverse immune responses to HBV surface epitope variants after vaccine booster in adolescents immunized in infancy.

OBJECTIVES: We aimed to investigate immunity against hepatitis B surface antigen (HBsAg) mutants before and after boosters in adolescents who had lost antibodies against HBsAg (anti-HBs) despite neonatal vaccination. METHODS: We recruited 142 participants from 15 to 21 years old who had received complete vaccination in infancy but became anti-HBs-negative. Cellular immunity to HBsAg and T-cell epitope variants was assessed before and after boosters. Antibody affinity to variants was assessed after boosters. RESULTS: After one dose of booster, 12 out of 140 (8.6%) participants remained anti-HBs-negative. Anti-HBs titres were higher in those participants <17 (geometric mean, 337.9 ± 10.9 vs. 157.4 ± 16.6 mIU/mL, p = 0.012). Before the booster, HBsAg-specific cell proliferation was present in 58 out of 64 (90.6%) participants. The proliferation response rates to T-cell epitopes were 37.8% and 72.6% (p < 0.001) before and after the booster, respectively. The stimulation index improved from 1.25 ± 1.70 to 2.53 ± 2.32 (p < 0.001) for various T-cell epitopes. HBsAg-specific interleukin (IL)-5- and interferon (IFN)-γ-secreting T-cells were enhanced (45 ± 10 and 50 ± 4 to 420 ± 328 and 355 ± 424 spot-forming cells/106 cells, respectively, p < 0.001). IFN-γ-secreting T cells to epitope 16-33 containing R24K and the antibody affinity to sG145R were still significantly lower than to the wild type. CONCLUSIONS: In immunized adolescents who lost anti-HBs, around 10% also lost immune memory. Cellular immunity to some T-cell epitope variants improved after the booster. Antibody affinity to sG145R and the IFN-γ-secreting cell response to some epitope 16-33 variants were still impaired even after booster administration.

Hepatocarcinogenesis in transgenic mice carrying hepatitis B virus pre-S/S gene with the sW172* mutation.

Hepatitis B virus (HBV) carrying the rtA181T/sW172* mutation conferred cross-resistance to adefovir and lamivudine. Cell-based and clinical studies indicated that HBV carrying this mutation had an increased oncogenic potential. Herein, we created transgenic mouse models to study the oncogenicity of the HBV pre-S/S gene containing this mutation. Transgenic mice were generated by transfer of the HBV pre-S/S gene together with its own promoter into C57B6 mice. Four lines of mice were created. Two of them carried wild-type gene and produced high and low levels of HBV surface antigen (HBsAg) (TgWT-H and L). The other two carried the sW172* mutation with high and low intrahepatic expression levels (TgSW172*-H and L). When sacrificed 18 months after birth, none of the TgWT mice developed hepatocellular carcinoma (HCC), whereas 6/26 (23.1%) TgSW172*-H and 2/24 (8.3%) TgSW172*-L mice developed HCC (TgWT vs TgSW172*; P=0.0021). Molecular analysis of liver tissues revealed significantly increased expression of glucose-regulated protein 78 and phosphorylated extracellular signal-regulated kinases 1 in TgSW172* mice, and decreased expression of B-cell lymphoma-extra large in TgSW172*-H mice. Higher proportion of apoptotic cells was found in TgSW172*-H mice, accompanied by increased cyclin E levels, suggesting increased hepatocyte turnover. Combined analysis of complimentary DNA microarray and microRNA array identified microRNA-873-mediated reduced expression of the CUB and Sushi multiple domains 3 (CSMD3) protein, a putative tumor suppressor, in TgSW172* mice. Our transgenic mice experiments confirmed that HBV pre-S/S gene carrying the sW172* mutation had an increased oncogenic potential. Increased endoplasmic reticulum stress response, more rapid hepatocyte turnover and decreased CSMD3 expression contributed to the hepatocarcinogenesis.

Hepatic expression of MxA and OAS1 in an ex vivo liver slice assay independently predicts treatment outcomes in chronic hepatitis C.

Antiviral effect of interferon is mediated by the expression of interferon-stimulated genes (ISGs). However, because of the difficulty in obtaining paired liver biopsies before and after interferon treatment, the key ISGs expressed in human hepatocytes and responsible for interferon-based antiviral activities in chronic hepatitis C remain illusive. Prior to a standard course of peginterferon plus ribavirin therapy, 104 patients underwent a liver biopsy. A small piece of the liver biopsy sample from each patient was submitted for ex vivo tissue culture in the presence or absence of interferon. Hepatic expression of 8 ISGs was detected by RT-PCR. The ISG expression patterns and clinicopathological variables were correlated with subsequent treatment outcomes. Multivariate logistic regression analysis showed that hepatic MxA expression (P = 0.008) and leucocyte count (P = 0.040) independently predicted the end of therapeutic virological response, while hepatic OAS1 expression (P = 0.003), genotype 1 (P = 0.002), HCV-RNA level (P = 0.007), AST/ALT ratio (P = 0.004) and leucocyte count (P = 0.034) independently predicted the sustained virological response. Immunohistochemistry analysis showed that interferon-induced OAS1 expression localized to the hepatocytes. In conclusion, hepatic MxA and OAS1 expression predicted, respectively, the end of therapeutic and sustained virological responses in interferon-based treatment of chronic hepatitis C.

Acyl-coenzyme A:cholesterol acyltransferase inhibition ameliorates proteinuria, hyperlipidemia, lecithin-cholesterol acyltransferase, SRB-1, and low-denisty lipoprotein receptor deficiencies in nephrotic syndrome.

BACKGROUND: Nephrotic syndrome (NS) is associated with hyperlipidemia, altered lipid regulatory enzymes and receptors, and increased risk of progressive renal and cardiovascular diseases. Acyl-coenzyme A:cholesterol acyltransferase (ACAT) catalyzes intracellular esterification of cholesterol and plays an important role in production of apolipoprotein B-containing lipoproteins, regulation of cholesterol-responsive proteins, and formation of foam cells. Because hepatic ACAT-2 is markedly upregulated in NS, we tested the hypothesis that inhibition of ACAT may improve cholesterol metabolism in NS. METHODS AND RESULTS: Rats with puromycin-induced NS were treated with either the ACAT inhibitor CI-976 or placebo for 2 weeks. Normal rats served as controls. Plasma lipids, renal function, and key lipid regulatory factors were measured. Untreated NS rats showed heavy proteinuria; hypoalbuminemia; elevated plasma cholesterol, triglyceride, LDL, VLDL, and total cholesterol-to-HDL cholesterol ratio; increased hepatic ACAT activity, ACAT-2 mRNA, and ACAT-2 protein; and reduced LDL receptor, HDL receptor, otherwise known as scavenger receptor B-1 (SRB-1) and plasma lecithin-cholesterol acyltransferase (LCAT). ACAT inhibitor reduced plasma cholesterol and triglycerides, normalized total cholesterol-to-HDL cholesterol ratio, and lowered hepatic ACAT activity without changing ACAT-2 mRNA or protein. This was accompanied by near normalizations of plasma LCAT, hepatic SRB-1, and LDL receptor and a significant amelioration of proteinuria and hypoalbuminemia. CONCLUSIONS: Pharmacological inhibition of ACAT reverses NS-induced LDL receptor, HDL receptor, and LCAT deficiencies; improves plasma lipid profile; and ameliorates proteinuria in nephrotic animals. Further studies are needed to explore the effect of ACAT inhibition in nephrotic humans.

Reversibility of chronic experimental syndrome X by diet modification.

This study was designed to examine whether abnormalities that comprise the metabolic syndrome, including insulin resistance, hyperinsulinemia, hypertension, hyperlipidemia, and obesity, are reversible by diet. Female Fischer rats were placed on either a high-fat, refined-carbohydrate (HFS) diet or low-fat, complex-carbohydrate (LFCC) diet for a period of 20 months. After 20 months, a group of HFS rats were switched to the LFCC diet (HFS/LFCC) for a period of 2 months. Skeletal muscle glucose transport, plasma insulin, systolic blood pressure, and plasma lipids were measured in all groups after 22 months. Energy intake and body weight were measured weekly. In the HFS group, insulin-stimulated glucose transport was significantly reduced (67+/-4 versus 98+/-4 pmol. mg(-)(1). 15 s(-)(1)), whereas plasma insulin (300+/-49 versus 82+/-8 pmol/L), blood pressure (147+/-4 versus 123+/-4 mm Hg), plasma triglycerides (2.58+/-0.31 versus 0.39+/-0.04 mmol/L), LDL cholesterol (C) (3.45+/-0.40 versus 0.89+/-0.06 mmol/L), LDL-C to HDL-C ratio (2.9+/-0.1 versus 2.2+/-0.1), VLDL-C (1.53+/-0.23 versus 0.37+/-0.07 mmol/l), Total-C (5.56+/-0.58 versus 1.49+/-0.10 mmol/L), and body weight (360+/-11 versus 260+/-5 g) were all significantly elevated compared with the LFCC. Energy intake did not differ significantly; however, the LFCC had a much poorer feed efficiency. Conversion to a LFCC diet for 2 months led to normalization of glucose transport, blood pressure, plasma insulin, and VLDL-C and significant amelioration of obesity and other lipid abnormalities. These results demonstrate that syndrome X induced by an inappropriate diet is reversed with implementation of a low-fat, unrefined-carbohydrate diet without caloric restriction and suggest that diet may be a possible treatment for multiple simultaneous cardiovascular risk factors.

Bounded diffusion for multiscale edge detection using regularized cubic B-spline fitting.

This paper shows that in edge detection the regularization factor alpha is a better scale parameter than the standard deviation (sigma) of the Gaussian pre-filter. The alpha scale space, which exhibits the evolutionary behaviour of an edge in various scales, is the basis for the design of a multiscale edge detector (MRCBS). In MRCBS, the scale is determined adaptively according to the local noise level; the thresholds which control the amount of edge details are adjusted according to the scale; and the anisotropic diffusion is applied in the finest scale to further suppress noise.

Effect of heparin on mesangial cell growth and gene expression of matrix proteins.

BACKGROUND: Mesangial cell (MC) proliferation and matrix expansion are characteristics of many glomerulopathies. Heparin has been shown to inhibit MC proliferation in vitro and mitigate cell proliferation, matrix expansion, proteinuria, renal insufficiency, and hypertension in experimental glomerulonephritis and subtotal renal ablation. We examined the effect of standard heparin on MC proliferation and matrix protein expression in vitro which necessarily excludes the confounding influences of haemodynamic, inflammatory, haemostatic, and various other processes that are present in vivo. METHODS: Gene expression and release of fibronectin (FN), collagen IV and laminin by cultured rat MC were tested in the presence and absence of heparin. In addition the effect of transforming growth factor-beta1 (TGF-beta1) on the gene expression of those matrix proteins was assessed. RESULTS: Within a 3-1000 microg/ml concentration range, heparin inhibited gene expression and release of FN by 10% fetal calf serum (FCS)-stimulated MC in a concentration-dependent manner. At concentrations of 300 and 1000 microg/ml, heparin inhibited fibronectin mRNA levels in TGF-beta1 (6 ng/ml) stimulated cells. However, heparin had no effect on gene expression or release of collagen IV or laminin under these conditions. Heparin markedly inhibited 10% FCS-stimulated MC proliferation in a concentration-dependent manner. CONCLUSIONS: Heparin inhibited MC growth and fibronectin production. These effects may, in part, account for the reported beneficial effects of heparin on the course of renal disease in experimental animals.

Down-regulation of hepatic LDL receptor expression in experimental nephrosis.

Nephrotic syndrome (NS) is invariably associated with elevation of plasma total and LDL cholesterol concentrations. The present study was carried out to test the hypothesis that nephrotic LDL hypercholesterolemia is, in part, due to acquired LDL receptor (LDLR) deficiency. To this end, hepatic LDLR mRNA (Northern blot analysis) and protein mass (Western blot analysis) were measured longitudinally before and during the course of puromycin-induced NS. In addition, the rate of LDLR gene transcription by isolated hepatic nuclei was determined using nuclear run-on assay. Hepatic LDLR mRNA remained virtually unchanged during the 30-day course of the study period. However, after an insignificant rise on day 5, LDLR protein mass gradually declined to a level which was significantly below the baseline values (P < 0.05 ANOVA). This was accompanied by a normal rate of LDLR mRNA synthesis excluding impaired gene transcription as a cause. The fall in hepatic LDLR protein was associated with a marked rise in plasma total and LDL cholesterol concentrations but no rise in hepatic tissue cholesterol concentration. The latter observation is indicative of impaired hepatic cholesterol uptake and provides functional evidence for the demonstrated acquired LDLR deficiency in the NS animals. Likewise, our findings elucidate the molecular basis of the previously reported impaired LDL clearance in NS. In conclusion, severe hypercholesterolemia in rats with experimental NS is associated with and perhaps, in part, is due to down-regulation of LDL receptor expression.

Gene expression of hepatic cholesterol 7 alpha-hydroxylase in the course of puromycin-induced nephrosis.

Cholesterol conversion to and biliary excretion of bile acids represents the principal pathway of cholesterol catabolism in mammals. Cholesterol 7 alpha-hydroxylase (Ch-7 alpha-H) is the first and the rate limiting step in bile acid production. Recently, Ch-7 alpha-H enzymatic activity has been shown to be normal in rats with established puromycin aminonucleoside-induced nephrosis (NS). To our knowledge, the gene expression of Ch-7 alpha-H in NS has not been investigated. We measured hepatic Ch-7 alpha-H mRNA and protein (by Northern and Western blot analyses) in rats at baseline and longitudinally during the course of induction and chronic phase of puromycin (PAN) induced NS. Groups of placebo-treated (controls) and diet-induced hypercholesterolemic (DHC) rats were included for comparison. The NS and DHC animals exhibited severe hypercholesterolemia of similar magnitude. Hepatic Ch-7 alpha-H transcript and protein remained virtually unchanged throughout the study period in the NS group. In contrast, Ch-7 alpha-H gene expression was markedly up-regulated in the DHC group. These observations suggest that hepatic Ch-7 alpha-H gene expression may be inappropriately low for the degree of the associated hypercholesterolemia in the NS group. It should be noted, however, that hepatic tissue cholesterol concentration was normal in the NS group and greatly increased in the DHC group. This can account for the disparity in Ch-7 alpha-H mRNA levels between the two groups since intracellular rather than extracellular cholesterol modulates Ch-7 alpha-H gene expression. In conclusion, the present study revealed that hepatic Ch-7 alpha-H gene expression remains unchanged during the course of PAN-induced NS in rats. It thus appears that generation and maintenance of hypercholesterolemia in this model of NS does not involve significant alteration of Ch-7 alpha-H gene expression.

Hepatic HMG-CoA reductase gene expression during the course of puromycin-induced nephrosis.

Increased production and depressed catabolism of lipoproteins play major roles in the pathogenesis of hypercholesterolemia of nephrotic syndrome (NS). However, the effect, if any, of NS on cholesterol biosynthetic capacity is uncertain. We examined the gene expression of hepatic 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoAR, the rate limiting step in cholesterol biosynthesis) during the induction and chronic phase of puromycin (PAN)-induced NS in rats. The rats were randomized to NS (given i.p. puromycin aminonucleoside 130 mg/kg on day 1 and 60 mg/kg on day 14) and placebo-treated control groups. Subgroups of animals were sacrificed at days 5, 10, 20 and 30. The liver was harvested between 7 and 9 p.m. for measurements of HMG-CoAR and actin mRNAs, HMG-CoAR enzymatic activity and microsomal cholesterol concentration. In separate experiments, subgroups of animals with chronic NS (day 30) were studied in fed and 20-hour fasting states. A marked but transient rise in hepatic HMG-CoAR mRNA and HMG-CoAR enzymatic activity was observed following the onset and exacerbation of proteinuria within a few days after each puromycin injection. On each occasion, HMG-CoAR fell to the baseline level despite persistent severe hypercholesterolemia. In an attempt to examine the possible acute effect of PAN per se, experiments were repeated before and at short intervals (8 and 24 hr) after puromycin injection when proteinuria was absent and the drug exposure prominent. The HMG-CoAR mRNA and activity were virtually unchanged during this period, suggesting the lack of an acute effect of puromycin. Twenty-hour fasting led to a marked rise in HMG-CoAR mRNA and activity in animals with chronic NS but not in the controls. Microsomal cholesterol remained unchanged and comparable in the two groups at all points. Thus, the marked but transient rise in hepatic HMG-CoAR gene expression observed during the induction phase and with fasting during the chronic phase of PAN-induced NS may contribute to the generation and maintenance of hypercholesterolemia in this animal model.

The effects of calcium-regulating-hormone on BMC in patients with CRF.

By means of single photon absorptiometry, bone material content (BMC) on centerbrachium was detected in 34 patients with chronic renal failure (CRF) and 34 sex-age-matched normal controls, and their serum parathyroid hormone (PTH), calcitonin (CT) were also measured by radioimmunoassay. It was found that (1) the mean BMC was obviously lower in patients with CRF than that in the normal controls; (2) PTH and CT obviously rose in patients with CRF; (3) the decrease of BMC was strongly correlated with that of creatinine clearance (Ccr). Furthermore, the more Ccr decreases, the more PTH in blood increases; no relations have been found between BMC and the serum levels of CT. The authors suggest that the development of osteoporosis with CRF is due to the secondary increase of PTH in blood but the effect of increased serum CT levels was not so obvious. Therefore, lowering blood PTH concentrations by parathyroidectomy or renal transplantation may be beneficial to curing osteoporosis of patients with CRF.

K-fold Crossvalidation in Canonical Analysis.

A computer-assisted, K-fold crossvalidation technique is discussed within the framework of canonical correlation analysis of randomly generated data sets. Results of the analysis suggest that this technique of multi-crossvalidation can be an effective method to reduce the contamination of canonical variates and canonical correlations by sample-specific variance components.

[Effect of radix Angelicae sinensis on serum gastrin levels in patients with cirrhosis].

Acute and chronic effect of Radix Angelicae Sinensis (RAS) on serum gastrin levels in patients with cirrhosis were investigated. The results showed that after intravenous perfusion of RAS, serum gastrin levels of inferior vana cava, hepatic and peripheral veins were significantly decreased. After long-term administration of the agent, the level fell nearly to that of control subjects. It is suggested that the effect of reducing serum gastrin level by RAS may improve portal hemodynamics and be beneficial for portal hypertensive gastroduodenal mucosal lesions in cirrhosis.

[A quantitative diagnosis of chronic gastritis].

To explore a definite diagnostic criterion of chronic gastritis, we undertook a quantitative study of the gastric biopsy specimens taken from 131 subjects. Seven stages were classified from normal gastric mucosa to mild, moderate and severe superficial and atrophic gastritis. The results shown the number of inflammatory cell was much greater in the mild superficial gastritis (128 +/- 84.37/HP) than in normal gastric mucosa (22 +/- 10.54HP, P less than 0.001). In these stages, the numbers of pyloric and fundic gland progressively decreased, the size of the gastric gland gradually enlarged and the percentage of intestinal metaplasia by degrees increased. According to these, we establish the quantitative diagnostic criterion of chronic gastritis.

[Effect of pituitrin or phentolamine alone or in combination on WHVP and systemic hemodynamics in patients with liver cirrhosis]. / Wirkung von Pituitrin und Phentolamin allein oder in Kombination auf WHVP und die systemische Hämodynamik bei Patienten mit Leberzirrhose.

We observed the effect of pituitrin and phentolamine alone or in combination on wedged hepatic venous pressure (WHVP) and systemic hemodynamics in 28 patients with cirrhosis. The results showed that either of these drugs used separately could lead to reduction in WHVP, each of them could also cause by-effects on systemic hemodynamics. When pituitrin in combination with phentolamine was administered, no change could be found in inferior vena cava pressure, mean arterial pressure, pulse rate and cardiac index. This suggested that pituitrin in combination with phentolamine could not only efficaciously decrease WHVP, but also counteract side effects on systemic hemodynamics of each other and improve hepatic microcirculation. Our study provided evidence for the usefulness of the combination of the two drugs in controlling bleeding from esophagus varices.