Negative regulation of signal transducer and activator of transcription-3 signalling cascade by lupeol inhibits growth and induces apoptosis in hepatocellular carcinoma cells

Siveen, K S,Nguyen, A H,Lee, J H,Li, F,Singh, S S,Kumar, A P,Low, G,Jha, S,Tergaonkar, V,Ahn, K S,Sethi, G Nature Publishing Group 2014 The British journal of cancer Vol.111 No.7

<P><B>Background:</B></P><P>Constitutive activation of signal transducer and activator of transcription signalling 3 (STAT3) has been linked with survival, proliferation and angiogenesis in a wide variety of malignancies including hepatocellular carcinoma (HCC).</P><P><B>Methods:</B></P><P>We evaluated the effect of lupeol on STAT3 signalling cascade and its regulated functional responses in HCC cells.</P><P><B>Results:</B></P><P>Lupeol suppressed constitutive activation of STAT3 phosphorylation at tyrosine 705 residue effectively in a dose- and time-dependent manner. The phosphorylation of Janus-activated kinases (JAKs) 1 and 2 and Src was also suppressed by lupeol. Pervanadate treatment reversed the downregulation of phospho-STAT3 induced by lupeol, thereby indicating the involvement of a phosphatase. Indeed, we observed that treatment with lupeol increased the protein and mRNA levels of SHP-2, and silencing of SHP-2 abolished the inhibitory effects of lupeol on STAT3 activation. Treatment with lupeol also downregulated the expression of diverse STAT3-regulated genes and decreased the binding of STAT3 to VEGF promoter. Moreover, the proliferation of various HCC cells was significantly suppressed by lupeol, being associated with substantial induction of apoptosis. Depletion of SHP-2 reversed the observed antiproliferative and pro-apoptotic effects of lupeol.</P><P><B>Conclusions:</B></P><P>Lupeol exhibited its potential anticancer effects in HCC through the downregulation of STAT3-induced pro-survival signalling cascade.</P>

Effect of Microbial Phytase on Performance, Nutrient Absorption and Excretion in Weaned Pigs and Apparent Ileal Nutrient Digestibility in Growing Pigs

Zeng, Z.K.,Piao, X.S.,Wang, D.,Li, P.F.,Xue, L.F.,Salmon, Lorraine,Zhang, H.Y.,Han, X.,Liu, L. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.8

Two experiments were conducted to evaluate the efficacy of Trichoderma reesei derived phytase for pigs fed diets with fixed calcium to total phosphorus ratios (1.5:1). In Exp. 1, 280 weaned pigs (initial BW of $10.32{\pm}1.94$ kg) were allocated to one of five dietary treatments on the basis of weight and gender in a randomized complete block design. Treatments were the low phosphorus (0.6% Ca, 0.4% total P and 0.23% available P) diets supplemented with 0, 250, 1,000, or 2,000 FTU phytase/kg of diet and a positive control diet (PC; 0.85% Ca, 0.58% total P and 0.37% available P). The treatments were applied to seven pens with eight pigs per pen, half male and half female. In Exp. 2, six barrows fitted with ileal T-cannula (initial BW = $35.1{\pm}1.6$ kg) were assigned to three dietary treatments with a double $3{\times}3$ Latin square design. The dietary treatments were the low-phosphorus diet (0.53% Ca, 0.34% total P and 0.14% available P), the low phosphorus diet plus 1,000 FTU phytase/kg and a positive control diet (0.77% Ca, 0.50% total P and 0.30% available P). In Exp. 1, there were linear increases (p<0.01) in weight gain, phosphorus absorption, bone strength, calcium and phosphorus content of fat-free dried bone and plasma phosphorus concentrations with increasing dose rate of phytase. The performance of pigs fed the diets with 250, 1,000, or 2,000 FTU of phytase/kg did not differ from pigs fed the PC diet. Pigs fed diets with 1,000 or 2,000 FTU of phytase/kg did not differ from pigs fed the PC diet in bone characteristics. The apparent digestibility of dry matter, crude protein, ash and energy was not affected by dietary treatment. However, pigs fed the PC diet excreted more fecal phosphorus (g/d, p<0.01) and fecal phosphorus per BW gain (g/kg) than pigs fed the diets with phytase. Phytase linearly decreased (p<0.01) fecal phosphorus excreted per BW gain (g/kg), plasma calcium concentration as well as plasma and bone alkaline phosphatase activity. In Exp. 2, phytase supplementation in the low-P diet increased (p<0.05) the apparent ileal digestibility (AID) of Ca, P, leucine, lysine, phenylalanine, alanine and cysteine, tended to AID of crude protein, isoleucine, threonine, asparagine and serine. In conclusion, the novel phytase originated from Trichoderma reesei is effective in releasing Ca, P, and amino acids from corn soy based diet for pigs.

Effects of Adding Super Dose Phytase to the Phosphorus-deficient Diets of Young Pigs on Growth Performance, Bone Quality, Minerals and Amino Acids Digestibilities

Zeng, Z.K.,Wang, D.,Piao, X.S.,Li, P.F.,Zhang, H.Y.,Shi, C.X.,Yu, S.K. Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.2

Two experiments were conducted to evaluate the efficacy of feeding an Escherichia coli (E. coli) derived phytase to pigs fed P deficient, corn-soybean meal diets. In Exp. 1, one hundred and twenty crossbred piglets ($9.53{\pm}0.84$ kg) were allocated to one of five treatments which consisted of four low P diets (0.61% Ca, 0.46% total P and 0.24% non-phytate P) supplemented with 0, 500, 1,000, or 20,000 FTU/kg E. coli phytase as well as a positive control formulated to be adequate in all nutrients (0.77% Ca, 0.62% total P and 0.42% non-phytate P). The treatments were applied to six pens with four pigs per pen for 28 days. In Exp. 2, ten crossbred pigs ($19.66{\pm}1.16$ kg) fitted with ileal T-cannula were used in a nutrient balance study. The pigs were assigned to treatments similar to those used in Exp. 1 in a doubly replicated $5{\pm}4$ incomplete Latin square design (5 diets with 4 periods). Each period consisted of a 5-d adjustment period followed by a 3-d total collection of feces and urine and then a 2-d collection of ileal digesta. Supplementation with phytase linearly increased (p<0.05) weight gain, feed intake, feed efficiency, bone breaking strength and fat-free dry and ash bone weight. There were linear increases (p<0.01) in the apparent ileal digestibility (AID) of DM, GE, CP, Ca, total P, inositol hexaphosphate ($IP_6$) and some AA with increasing dose of E. coli phytase. Pigs fed 20,000 FTU/kg had a greater (p<0.05) AID of IP6 (80% vs 59% or 64%, respectively) than pigs fed diets with 500 or 1,000 FTU/kg phytase. There were linear increases (p<0.05) in the total tract digestibility of Ca, total P, Na, K, Mg, and Zn as well as in the retention of Mg and Zn with increased phytase dose. The retention and utilization of Cu, and the total tract digestibility of CP and Cu quadratic increased (p<0.05) with increased phytase dose. In conclusion, supplementation of 500 FTU of phytase/kg and above effectively hydrolyzed phytate in low-P corn-soybean diets for pigs. In addition, a super dose of phytase (20,000 FTU/kg) hydrolyzed most of the IP6 and consequently further improved mineral use, protein utilization and performance.

Effects of Fermented Potato Pulp on Performance, Nutrient Digestibility, Carcass Traits and Plasma Parameters of Growing-finishing Pigs

Li, P.F.,Xue, L.F.,Zhang, R.F.,Piao, Xiangshu,Zeng, Z.K.,Zhan, J.S. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.10

A total of 629 Duroc${\times}$Landrace${\times}$Large White crossbred pigs were utilized in three experiments (Exp. 1, 222 pigs weighing $25.6{\pm}2.0\;kg$ BW; Exp. 2, 216 pigs weighing 5$6.2{\pm}4.3\;kg$ BW; Exp. 3, 191 pigs weighing $86.4{\pm}4.6\;kg$ BW) conducted to determine the effects of fermented potato pulp on performance, nutrient digestibility, carcass traits and plasma parameters in growingfinishing pigs. Each experiment lasted 28 d. The pigs were assigned to one of two corn-soybean meal-based diets containing 0 or 5% fermented potato pulp. The inclusion of fermented potato pulp increased weight gain (p<0.05) in experiments 1 and 2 and increased feed intake (p<0.05) in experiment 2. Feed conversion was improved (p<0.05) in experiment 2 and showed a tendency to improve (p<0.10) in experiments 1 and 3 when pigs were fed fermented potato pulp. Fermented potato pulp increased (p<0.05) dry matter digestibility in experiments 1 and 3 and energy digestibility in experiment 2. Feeding fermented potato pulp decreased plasma urea nitrogen (p<0.05) and alanine aminotransferase (p<0.05) in experiments 1 and 2, while plasma aspartate aminotransferase was decreased (p<0.05) in experiment 3. Dietary fermented potato pulp did not affect the carcass characteristics of finishing pigs. Feeding fermented potato pulp reduced (p<0.05) fecal ammonia concentration in all three experiments. In conclusion, feeding growing-finishing pigs diets containing 5% fermented potato pulp improved weight gain and feed conversion without any detrimental effects on carcass traits. The improvements in pig performance appeared to be mediated by improvements in nutrient digestibility.

Genome-wide association study in a Chinese population identifies a susceptibility locus for type 2 diabetes at 7q32 near <i>PAX4</i>

Ma, R. C. W.,Hu, C.,Tam, C. H.,Zhang, R.,Kwan, P.,Leung, T. F.,Thomas, G. N.,Go, M. J.,Hara, K.,Sim, X.,Ho, J. S. K.,Wang, C.,Li, H.,Lu, L.,Wang, Y.,Li, J. W.,Wang, Y.,Lam, V. K. L.,Wang, J.,Yu, W.,Ki Springer-Verlag 2013 Diabetologia Vol.56 No.6

<P><B>Aims/hypothesis</B></P><P>Most genetic variants identified for type 2 diabetes have been discovered in European populations. We performed genome-wide association studies (GWAS) in a Chinese population with the aim of identifying novel variants for type 2 diabetes in Asians.</P><P><B>Methods</B></P><P>We performed a meta-analysis of three GWAS comprising 684 patients with type 2 diabetes and 955 controls of Southern Han Chinese descent. We followed up the top signals in two independent Southern Han Chinese cohorts (totalling 10,383 cases and 6,974 controls), and performed in silico replication in multiple populations.</P><P><B>Results</B></P><P>We identified <I>CDKN2A/B</I> and four novel type 2 diabetes association signals with <I>p</I> < 1 × 10<SUP>−5</SUP> from the meta-analysis. Thirteen variants within these four loci were followed up in two independent Chinese cohorts, and rs10229583 at 7q32 was found to be associated with type 2 diabetes in a combined analysis of 11,067 cases and 7,929 controls (<I>p</I><SUB>meta</SUB> = 2.6 × 10<SUP>−8</SUP>; OR [95% CI] 1.18 [1.11, 1.25]). In silico replication revealed consistent associations across multiethnic groups, including five East Asian populations (<I>p</I><SUB>meta</SUB> = 2.3 × 10<SUP>−10</SUP>) and a population of European descent (<I>p</I> = 8.6 × 10<SUP>−3</SUP>). The rs10229583 risk variant was associated with elevated fasting plasma glucose, impaired beta cell function in controls, and an earlier age at diagnosis for the cases. The novel variant lies within an islet-selective cluster of open regulatory elements. There was significant heterogeneity of effect between Han Chinese and individuals of European descent, Malaysians and Indians.</P><P><B>Conclusions/interpretation</B></P><P>Our study identifies rs10229583 near <I>PAX4</I> as a novel locus for type 2 diabetes in Chinese and other populations and provides new insights into the pathogenesis of type 2 diabetes.</P><P><B>Electronic supplementary material</B></P><P>The online version of this article (doi:10.1007/s00125-013-2874-4) contains peer-reviewed but unedited supplementary material, which is available to authorised users.</P>

Effects of Keratinase on Performance, Nutrient Utilization, Intestinal Morphology, Intestinal Ecology and Inflammatory Response of Weaned Piglets Fed Diets with Different Levels of Crude Protein

Wang, D.,Piao, X.S.,Zeng, Z.K.,Lu, T.,Zhang, Q.,Li, P.F.,Xue, L.F.,Kim, S.W. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.12

Two experiments were conducted to investigate the in vitro ability of keratinase to hydrolyze soybean glycinin and ${\beta}$-conglycinin and to evaluate the in vivo effects of keratinase when included in corn-soybean diets with different levels of crude protein and fed to nursery pigs. In experiment 1, a saturated keratinase solution (1 ml) was added to two blank controls of either glycinin or ${\beta}$-conglycinin resulting in the hydrolysis of 94.74% glycinin and 88.89% ${\beta}$-conglycinin. In experiment 2, 190 pigs (8.3${\pm}$0.63 kg BW) were allotted to one of four treatments in a 2${\times}$2 factorial arrangement on the basis of body weight, and sex was balanced among the pens. The effects of crude protein (19 vs. 22%) and keratinase (0 vs. 0.05%) were studied. Each treatment was applied to six pens with seven (two pens) or eight pigs per pen. Pigs were fed the experimental diets for 21 d. Weight gain and feed conversion ratio were improved (p<0.05) with keratinase supplementation while feed intake was reduced (p<0.05). Keratinase supplementation increased (p<0.05) the apparent total tract digestibility of dry matter, energy, crude protein and phosphorus. Keratinase supplementation also increased n-butyric acid in the cecum and colon, lactobacilli and total anaerobe counts in the colon as well as the ratio of villus height to crypt depth in the ileum. Additionally, fecal score, ammonia nitrogen and branch chain volatile fatty acids in the colon, E. coli and total aerobe counts in the colon, crypt depth in the jejunum and ileum as well as serum interleukin-1 and interleukin-6 concentrations were also decreased (p<0.05) by keratinase supplementation. A reduction in dietary crude protein decreased (p<0.05) colon ammonia nitrogen concentration and cecal propionic acid and branch chain volatile fatty acid concentrations. In addition, cecal E. coli counts, colon total anaerobe counts, ileal crypt depth, and serum interleukin-1 and interleukin-6 concentrations were also decreased (p<0.05) with the reduction of dietary crude protein. With the exception of fecal scores, there were no significant interactions between crude protein and keratinase. This study provides evidence that dietary keratinase supplementation improved nursery pig performance by improving intestinal morphology and ecology, thus improving nutrient digestibility and alleviating the inflammatory response.

Effects of Dietary Electrolyte Balance on Growth Performance, Nitrogen Metabolism and Some Blood Biochemical Parameters of Growing Rabbits

Li, J.W.,Wang, X.P.,Wang, C.Y.,Zhu, Y.L.,Li, F.C. Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.12

The effects of different dietary electrolyte balance (DEB) on growth performance, nitrogen (N) metabolism and some blood biochemical parameters were investigated in 2 to 3 months old growing rabbits. A total of 150 growing rabbits of 2 months age were randomly divided into five groups according to average body weight, with 30 rabbits in each group. The DEB levels of the five experimental diets were -154, -3.16, +201, +347, and +500 meq/kg of dry matter (DM), respectively. There was a 7-d adaptation period and a 23-d experimental period. The results showed that the DEB levels had a quadratic affect on the average daily feed intake (ADFI) (p<0.001). The greatest ADFI was achieved when the DEB level was +201 meq/kg DM. Fecal N (FN) content linearly decreased (0.047), while digestible N (DN), retained N (RN), efficiency of intake N converted into digestible N (DN/IN) and the efficiency of intake N converted into retained N (RN/IN) linearly increased with the DEB increase (0.020, 0.004, 0.021, and 0.049, respectively). Serum phosphorus (P) ion content linearly increased with the DEB increase (p = 0.036). The DEB had a quadratic relationship with serum anion gap (AG) (p = 0.002) and serum parathyroid hormone (PTH) content (p = 0.016). The DEB levels quadratically affected base excess (BE) in the plasma (p<0.001). In conclusion, the DEB unaffected growth performance but affected feed intake, N metabolism and some blood biochemical parameters of growing rabbits.

Growth Performance, Meat Quality and Fatty Acid Metabolism Response of Growing Meat Rabbits to Dietary Linoleic Acid

Li, R.G.,Wang, X.P.,Wang, C.Y.,Ma, M.W.,Li, F.C. Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.8

An experiment was conducted to determine the effects of different amounts of dietary linoleic acid (LA) on growth performance, serum biochemical traits, meat quality, fatty acids composition of muscle and liver, acetyl-CoA carboxylase (ACC) and carnitine palmitoyl transferase 1 (CPT 1) mRNA expression in the liver of 9 wks old to 13 wks old growing meat rabbits. One hundred and fifty 9 wks old meat rabbits were allocated to individual cages and randomly divided into five groups. Animals in each group were fed with a diet with the following LA addition concentrations: 0, 3, 6, 9 and 12 g/kg diet (as-fed basis) and LA concentrations were 0.84, 1.21, 1.34, 1.61 and 1.80% in the diet, respectively. The results showed as follows: the dietary LA levels significantly affected muscle color of LL included $a^*$ and $b^*$ of experimental rabbits (p<0.05). The linear effect of LA on serum high density lipoprotein cholesterol was obtained (p = 0.0119). The saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs) contents of LL decreased and the polyunsaturated fatty acids (PUFAs) content of LL increased with dietary LA increase (p<0.0001). The PUFA n-6 content and PUFA n-3 content in the LL was significantly affected by the dietary LA levels (p<0.01, p<0.05). The MUFAs content in the liver decreased and the PUFAs contents in the liver increased with dietary LA increase (p<0.0001). The PUFA n-6 content and the PUFA n-6/n-3 ratio in the liver increased and PUFA n-3 content in the liver decreased with dietary LA increase (p<0.01). The linear effect of LA on CPT 1 mRNA expression in the liver was obtained (p = 0.0081). In summary, dietary LA addition had significant effects on liver and muscle fatty acid composition (increased PUFAs) of 9 wks old to 13 wks old growing meat rabbits, but had little effects on growth performance, meat physical traits and mRNA expression of liver relative enzyme of experimental rabbits.

Effect of Feeding Organic Acid With or Without Enzyme on Intestinal Microflora, Intestinal Enzyme Activity and Performance of Weaned Pigs

Li, Defa,Liu, S.D.,Qiao, S.Y.,Yi, G.F.,Liang, C.,Thacker, P. Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.3

Ninety-six, 35 day old, crossbred pigs, were fed either a basal diet based on com, soybean meal, fish meal and whey or one of three similar diets supplemented with either 0.5% organic acid or enzyme both alone and in combination. Neither organic acid nor enzyme produced any significant (p<0.05) improvements in daily gain or feed conversion. Organic acid, both alone and in combination with enzyme, significantly (p=0.04) decreased the pH in the lower colon. None of the treatments produced any effects on E. Coli or Lactobacillus numbers in any part of the gastrointestinal tract. Feeding enzyme increased trypsin (p=0.01), chymotrypsin (p=0.03) and amylase (p=0.08) levels in the jejunum. Chymotrypsin levels were higher (p=0.04) in the ileum of pigs fed enzyme. Serum glucose levels were lower (p=0.01) on day 14 in pigs fed enzyme either alone or in combination with acid. Under the conditions of this experiment (10% dietary whey, pigs weaned at 35 days of age), neither organic acid nor enzyme were effective in improving starter pig performance. Therefore, there would appear to be little justification for the routine inclusion of these products in diets fed to pigs weaned at 35 days or later.

Intramuscular Administration of Zinc Metallothionein to Preslaughter Stressed Pigs Improves Anti-oxidative Status and Pork Quality

Li, L.L.,Hou, Z.P.,Yin, Y.L.,Liu, Y.H.,Hou, D.X.,Zhang, B.,Wu, G.Y.,Kim, S.W.,Fan, M.Z.,Yang, C.B.,Kong, X.F.,Tang, Z.R.,Peng, H.Z.,Deng, D.,Deng, Z.Y.,Xie, M.Y.,Xiong, H.,Kang, P.,Wang, S.X. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.5

This study was conducted to determine the effects of exogenous zinc-metallothionein (Zn-MT) on anti-oxidative function and pork quality. After feeding a corn-soybean meal-based diet for two weeks, 48 pigs ($Duroc{\times}Landrace{\times}Chinese\;Black Pig$) were assigned randomly to four groups. Pigs in Group 1 were maintained under non-stress conditions, whereas pigs in Groups 2, 3 and 4 were aggressively handled for 25 min to produce stress. Pigs in Groups 1, 2, 3, and 4 received intramuscular administration of saline (control group; CON), 0 (negative control group; NCON), 0.8 (low dose group; LOW), and 1.6 (high dose group; HIGH) mg rabbit liver Zn-MT per kg body weight, respectively. Pigs were slaughtered at 3 and 6 h post-injection. Zn-MT treatment increased (p<0.05) the activities of superoxide dismutase (SOD) and glutathione-peroxidase (GSH-PX) while decreasing the concentration of malondialdehyde (MDA) in liver. These responses were greater (p<0.05) at 6 h than at 3 h post Zn-MT injection. Zn-MT treatment increased (p<0.05) hepatic SOD mRNA levels in a time and dose-dependent manner and decreased (p<0.05) serum glutamate-pyruvate transaminase and lactate dehydrogenase activities (indicators of tissue integrity). Zn-MT administration decreased (p<0.05) lactate concentration and increased (p<0.05) pH and water-holding capacity in the longissimus thorasis meat. Collectively, our results indicate that intramuscular administration of Zn-MT to pre-slaughter stressed pigs improved tissue anti-oxidative ability and meat quality.