Expression and prognostic significance of cyclin D1, RBL2/p130 and MCM7 in hepatocellular carcinoma

AIM: To investigate the expression and prognostic significance of cyclin D1, retinoblastoma-like protein 2(RBL2/p130) and minichromosome maintenance protein 7(MCM7) in hepatocellular carcinoma(HCC). METHODS: The expression of cyclin D1, RBL2/p130 and MCM7 in 44 HCC specimens, 26 adjacent noncancerous cirrhotic liver specimens and 18 normal liver specimens were detected by the method of immunohistochemistry. The correlations of cyclin D1, RBL2/p130 and MCM7 with the clinical parameters of HCC patients were analyzed. RESULTS: The positive expression rates of cyclin D1 and MCM7 in HCC were 68.2% and 72.7%, higher than those in normal livers and adjacent noncancerous cirrhotic livers(P<0.01). The positive expression rate of RBL2/p130 in HCC was 34.1%, lower than that in normal livers and adjacent noncancerous cirrhotic livers(P<0.01). The expression of MCM7 in HCC was positively correlated with the expression of cyclin D1(r=0.349, P<0.05), and negatively correlated with the expression of RBL2/p130(r=-0.421, P<0.01). The expression of cyclin D1 in HCC was negatively correlated with the expression of RBL2/p130(r=-0.435, P<0.01). The expression of MCM7 and cyclin D1 was associated with the integrity of tumor capsule, differentiation degree and TNM stage(P<0.05). The expression of MCM7 was also associated with tumor size and the value of alpha fetoprotein(AFP). The tumor size, the expression levels of MCM7 and cyclin D1 were correlated with the prognosis determined by multivariable analysis. The patients with positive expression of MCM7 and cyclin D1 had lower survival rate than those with negative expression(P<0.05). The patients with positive expression of RBL2/p130 had higher survival rate than those with negative expression(P<0.05). CONCLUSION: The abnormal expression of cyclin D1, RBL2/ p130 and MCM7 plays an important role in the development of HCC, indicating that monitoring their expression in HCC patients may be helpful to the judgment of prognosis.     

Monocrotophos augments the early alterations in lipid profile and organ toxicity associated with experimental diabetes in rats

The present study was undertaken to investigate the potential of monocrotophos (MCP), a widely used organophosphorus insecticide, to induce hyperglycemia and its interactive ability to accentuate the early diabetic outcomes and associated complications in experimentally rendered diabetic rats. Rats were rendered diabetic with an acute dose (60 mg/kg b.w, i.p) of streptozotocin. MCP was orally administered at a sublethal dose (1/20 LD_50, 0.9 mg /kg b.w./d, 5 days) to both normal and diabetic rats. While MCP per se moderately increased (by 25%) the blood glucose levels in normal rats, it significantly aggravated the hyperglycemic outcome in diabetic rats (56% above diabetic rats). Further, experimentally-induced diabetes was associated with only a marginal increase in total and HDL-cholesterol levels, while serum triglyceride levels were significantly enhanced. Although MCP per se did not affect these parameters, it caused a marked increase in triglyceride levels in serum of diabetic rats (54% above diabetic control). Furthermore, MCP-induced higher activity levels of serum transaminases viz., ALT and AST (51% and 71% higher as compared to diabetic control) suggestive of enhanced hepatic damage. Collectively, our findings provide evidence that MCP on repeated exposure has the propensity to augment the secondary complications associated with diabetes in a rat model.     

朗德鹅与籽鹅、莱籽鹅杂交的产肝性能观察

随机选择朗德鹅与籽鹅、莱籽鹅的杂交鹅雏各50只,待养至4月龄后达成年体重时,每组选择20只,进行为期3～4周的填饲试验。测量填饲肥肝平均重、平均耗料量和肝料比。结果表明：朗德×菜籽鹅的平均肝重为669．00g,极显著高于朗德×籽鹅（P〈0．01）,较朗德×籽鹅、菜籽鹅与籽鹅分别提高了163．09g、289．00g和365．85g,即分别提高了32．24％、76．05％和120．68％;同时降低了单位肥肝的饲料消耗量,肝料比为1：33．71。因此,从肥肝重和耗料上综合评价得出,朗德×菜籽鹅组的填饲效果最佳,是生产肥肝最好的杂交组合。     

Astragaloside Ⅳ inhibits JAK2/STAT3 signaling pathway and alleviates severe acute pancreatitis-associated acute liver injury in rats

AIM: To investigate the effect of astragaloside Ⅳ on severe acute pancreatitis (SAP)-associated acute liver injury in the rats and to explore the underlying mechanisms. METHODS: Male Sprague-Dawley (SD) rats (n=96) were randomly divided into sham-operated group, SAP model group, astragaloside Ⅳ treatment group and AG490 treatment group. SAP model was induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the biliopancreatic duct. The rats in astragaloside Ⅳ treatment group were intraperitoneally injected with 20 mg/kg astragaloside Ⅳ, while the rats in AG490 treatment group were injected with 8.0 mg/kg AG490 2 h before sodium taurocholate injection. The rats in sham-operated group and model group received the same volume of saline. The rats were sacrificed at 12 h, 18 h and 24 h after the treatment. The levels of ascites, serum amylase, ALT and AST were detected after the blood samples were collected by the puncture through inferior vena cava. The serum levels of TNF-α, IL-6 and IL-1β were also examined by ELISA. Furthermore, HE staining was used to observe the liver pathological changes, and the protein levels of p-JAK2 and p-STAT3 in the liver were evaluated by Western blot. RESULTS: Compared with sham-operated group, the levels of ascites, serum amylase, ALT, AST, IL-6, TNF-α and IL-1β in the rats in model group were significantly increased, while they were decreased in the rats in astragaloside Ⅳ treatment group and AG490 treatment group compared with the rats in model group. Meanwhile, the phosphorylation levels of JAK2 and STAT3 was significantly increased in model group compared with sham-operated group. The rats in astragaloside Ⅳ treatment group and AG490 treatment group both had a better improvement in the liver injury and lower phosphorylation levels of JAK2 and STAT3.CONCLUSION: Astragaloside Ⅳ exerts a protective effect on pancreatitis-associated acute liver injury in the rats possibly via inhibiting JAK2/STAT3 signaling pathway.     

Alterations in enzyme activities in vital organs of triploid female catfish Heteropneustes fossilis (Bloch)

Triploid Heteropneustes fossilis (Bloch) showed sterility and higher growth potential than the normal diploid fish. Activities of some metabolic enzymes such as cytosolic NADP-malate dehydrogenase (NADP-MDH), mitochondrial NAD-malate dehydrogenase (NAD-MDH) and glutamate pyruvate transaminase (GPT) were evaluated in liver, brain and kidney along with glucose-6-phosphate dehydrogenase (G-6-P D) in ovary of female triploid catfish. Activities of these enzymes showed distinct seasonal periodicity, mostly with highest activities in prespawning and spawning periods, in both diploid and triploid catfish but differed in magnitude. In triploid liver, GPT showed higher activity than the diploid counterpart in prespawning and spawning periods. On the contrary, mitochondrial NAD-MDH and cytosolic NADP-MDH in this organ showed a consistent lower activity than the diploid in all stages or in some stages of reproductive cycle respectively. Interestingly, none of the enzymes in brain and kidney of triploid female catfish showed significant changes in comparison to the diploid counterpart. The triploid ovary maintained a significantly lower level of G-6-P D activity throughout the resting, preparatory and pre-spawning periods compared to the diploid ovary. Lower level of malic enzymes (NAD-MDH and NADP-MDH) in liver and G-6-P D in ovary are in close synchrony with lower level of estradiol-17β in plasma of female triploids as found in earlier study.     

Utilization of Dietary β-Carotene and Retinol Acetate by Goslings and Young Geese

One-day-old and 3-month-old geese were fed different concentrations of g -carotene (5, 10, 50 or 150 mg kg -1 feed) or retinol acetate (10 000, 20 000 or 40 000 IU kg -1 feed) for about 3 weeks to assess their ability to convert g -carotene to vitamin A. Blood serum concentrations of g -carotene, retinol, f -tocopherol, triacylglycerides and cholesterol, as well as liver concentrations of g -carotene, retinol, triacylglycerides and cholesterol, were included as response factors. g Carotene was not detectable in serum and liver. Serum and liver retinol concentrations were positively correlated with dietary concentrations of g -carotene and retinol acetate ( R 2 = 0.84 and 0.95, respectively). In goslings 1 mg of g -carotene corresponded to 63 IU retinol and in young geese to 1216 IU, equivalent to 3.8% and 72.9%, respectively, of the theoretical value of 1667 IU retinol. Cholesterol concentrations in blood serum and liver were not affected ( P > 0.05) by the dietary levels of g -carotene and retinol acetate. The concentration of f -tocopherol in blood serum decreased with increasing dietary levels of g -carotene and retinol acetate. In the goslings the concentration of triacylglycerides in serum and liver was significantly ( P < 0.05) influenced by the intake of g -carotene, but not by the intake of retinol acetate. In the young geese the serum concentration of triacylglycerides decreased ( P < 0.05) with increasing dietary levels of g -carotene, but was not affected by the dietary concentrations of retinol acetate. The concentration of triacylglycerides in liver tissue of young geese was not affected by the dietary concentration of either g -carotene or retinol acetate ( P > 0.05).     

Effects of Different Fat Supplements on Growth and Hepatic Lipids and Fatty Acids in Male Mink

Male mink kits (n=10 for each group) were fed diets supplemented with different fats for 12 weeks (September-November). The levels of digestible fat (8%) and energy content (7 MJ kg-1) of the diets were equal. The supplements used were beef pork fat, mink fat, broiler offal, rainbow trout offal, capelin oil, soybean oil and linseed oil. The growth and hepatic lipids (analysed by a thin-layer chromatography- flame ionization detection analyser) and fatty acid composition (analysed by gas-liquid chromatography) were studied. The pattern of weight gain of the mink fed the beef pork diet differed from that of the other mink. These kits reached high but delayed weight maxima compared with the other mink but then during November they lost weight rapidly. In liver, both the capelin oil- and linseed oil-fed mink had large concentrations of total lipids and triacylglycerols. The mink fed capelin oil were significantly heavier. The fatty acid analyses of hepatic total lipids and phospholipids revealed that the f -linolenic acid (18:3 n-3) of linseed oil was not efficiently metabolized to longer chain and more unsaturated fatty acids important for cellular membranes. It is discussed that 18:3 n-3 may not be as valuable for growing mink kits as the polyunsaturated fatty acids of the fish oils.     

Ischemic preconditioning protects against hepatic ischemia-reperfusion injury by attenuating leukotriene C4 production

AIM：To investigate the effect of ischemic preconditioning (IP) on leukotriene C4 (LTC4) generation during hepatic ischemia-reperfusion(I/R) injury.METHODS：Adult male Sprague-Dawley rats were randomly divided into sham (control) group, I/R group and IP+I/R group. The rat liver was subject to 60 min of partial hepatic ischemia followed by 5 h of reperfusion. LTC4 content was measured by reversed-phase high-performance liquid chromatography (RP-HPLC). The activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was determined, and the changes of histological structures were observed to assess the tissue injury. The reduced glutathione (GSH) level in the liver tissues was also examined by biochemical methods.RESULTS：IP markedly decreased LTC4 content in rat livers compared with I/R group. The activity of serum ALT and AST in I/R group and IP+I/R group was higher than that in control group. Compared with I/R group, the levels of serum ALT and AST were markedly decreased, and the level of GSH in hepatic tissues was elevated in IP+I/R group. In addition, the structural damage of the rat liver tissues in IP+I/R group displayed slighter than that in I/R group.CONCLUSION：IP treatment reduces LTC4 production accompanied with improving the liver function and histological structure during I/R injury, suggesting that the beneficial effects of IP may be involved in its repressing LTC4 generation during hepatic I/R injury.     

Glucose-regulated protein 78 promote cardiomyocyte apoptosis in liver cirrhotic rats

AIM: To explore the cardiomyocyte apoptosis induced by glucose-regulated protein 78/immunoglobulin heavy chain-binding protein (GRP78/BiP) in liver cirrhotic rats and its mechanism. METHODS: The liver cirrhotic rat model was established with multiple pathogenic factors, and sampled at the time points of 4 weeks, 6 weeks and 8 weeks. In experiment 1, the heart was collected and weighed, the thickness of the left ventricular wall was measured, and the ratios of the left ventricular wall thickness to the heart weight, and the heart weight to the body weight were calculated. In experiment 2, TUNEL was used to detect the apoptotic cardiomyocytes,and the protein levels of GRP78/BiP, CCAAT/enhancer-binding protein homologous protein/growth arrest and DNA damage-inducible protein 153 (CHOP/GADD153), caspase-12, nuclear factor-κB p65 (NF-κB p65) and B-cell lymphoma/leukemia-2 (Bcl-2) in the myocar-dium were detected by the method of immunohistochemistry. RESULTS: Compared with the normal myocardium, significant larger ratios of the left ventricular wall thickness to the heart weight and the heart weight to the body weight, a significant increase in the cardiomyocyte apoptosis, and a significant larger positive expression index of GRP78/BiP in the hearts 8 weeks after modeling were observed. The protein levels of CHOP/GADD153 and caspase-12 were gradually increased during the development of liver cirrhosis and were significantly increased at 8 weeks. The positive expression of NF-κB p65 and Bcl-2 showed consistent changes, and were markedly higher at 4 weeks than those at the other time points. The positive expression index of GRP78/BiP was positively correlated with the apoptotic index, and the levels of CHOP/GADD153 and caspase-12. The positive expression index of CHOP/GADD153 was negatively correlated with NF-κB p65 and Bcl-2. CONCLUSION: Elevated expression of GRP78/BiP may play a crucial role in the pathogenesis of liver cirrhotic cardio-myopathy mediated by endoplasmic reticulum stress.