Shu-Ling Hsieh

Immune response of white shrimp, Litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus

In the present study, we investigated immunological changes in viral-infected white shrimp, Litopenaeus vannamei. White shrimp were infected with white spot syndrome virus (WSSV) or co-infected with WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV) as detected by polymerase chain reaction (PCR). The complete (100%) mortality rate of shrimp was caused by viral infection due to immune parameters being suppressed including decreases in phenoloxidase activity, total hemocyte counts, differential hemocyte counts, and the gene expressions of prophenoloxidase and peroxinectin. In addition, increases in lipopolysaccharide and beta-1,3-glucan-binding protein of hemocytes and the hepatopancreas, and respiratory bursts per cell, and a decrease in superoxide dismutase were found in viral-infected shrimp, which may have been related to the defense against viral infection.

The increase of immunity and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii by feeding with selenium enriched-diet.

The effects of inorganic selenium (Se) (sodium selenate, SSe) and organic selenium (seleno-l-methionine, MSe) supplementation on the immune response, antioxidant status, and disease resistance of the giant freshwater prawn, Macrobrachium rosenbergii, were studied. Five experimental diets, including a control diet (without Se enrichment), 0.5 mg (kg diet)(-1) of MSe, 1 mg (kg diet)(-1) of MSe, 0.5 mg (kg diet)(-1) of SSe, and 1 mg (kg diet)(-1) of SSe, were used. After 75 days of culture, prawn fed the Se-enriched diets had lower mortality compared to that of prawn fed the control diet after being challenged by the pathogen, Debaryomyces hansenii. No significant differences in the total hemocyte count, superoxide dismutase activity, or clearance efficiency of prawn were recorded among the control and treated groups. Significantly increased phenoloxidase and phagocytic activities in prawn fed the Se-enriched diets were found compared to the controls. Respiratory bursts of prawn fed both forms of 1 mg Se (kg diet)(-1) significantly increased compared to control prawns. For the antioxidant status analysis, glutathione peroxidase, glutathione reductase, and glutathione s-transferase of prawn fed the SSe-enriched diet at 1 mg (kg diet)(-1) were significantly increased. The results indicated that the cheaper selenium, SSe is recommended to be added in prawn feed at the concentration of 0.5 mg resulting in 1.5 mg SSe (kg diet)(-1) increased prawn immunity and disease resistance against the pathogen, D. hansenii.

Effects of hot-water extract of Toona sinensis on immune response and resistance to Aeromonas hydrophila in Oreochromis mossambicus

Respiratory burst, lysozyme and phagocytic activities, and immunoglobulin levels in response to the pathogen Aeromonas hydrophila were examined in tilapia (Oreochromis mossanbicus, 10.65 +/- 2.5 g) injected individually with hot-water extract of Toona sinensis at 4 or 8 microg g(-1). Tilapia receiving the hot-water extract of T. sinensis at either dose had significantly increased respiratory burst, phagocytic activity and lysozyme activity towards A. hydrophila by 1 and 2 days post injection. No significant differences in total immunoglobulin levels were observed among the tilapia that received the two different doses of hot-water extract of T. sinensis at 4 and 8 microg g(-1). In another experiment, a Tilapia that had been injected with hot-water extract of T. sinensis was challenged with A. hydrophila at 5 x 10(7) colony-forming units (cfu) fish(-1). The survival of tilapia that received the hot-water extract of T. sinensis at 8 microg g(-1) was significantly higher than fish that received phosphate buffered saline and the control fish after 2 days, and at the termination of the experiment (7 days after the challenge). It was concluded that the hot-water extract of T. sinensis at 8 microg g(-1) or less had increased the immune response and resistance to A. hydrophila infection in tilapia.

Effects of Rutin from Toona sinensis on the immune and physiological responses of white shrimp (Litopenaeus vannamei) under Vibrio alginolyticus challenge.

Rutin is a bioflavonoid with strong antioxidant activity. To investigate the regulatory roles of rutin in various functions in crustaceans, we examined physiological (haemolymph glucose, lactate, and lipid) and innate non-specific immune responses (total haemocyte count (THC), phenoloxidase activity (PO), respiratory bursts (release of superoxide anion, O(2)(-)) and superoxide dismutase (SOD) activity) to the pathogen Vibrio alginolyticus in white shrimp (Litopenaeus vannamei) that were individually injected with rutin extracted from Toona sinensis at 10, 20, or 50 microg g(-1). Results showed that PO activity and respiratory burst of L. vannamei increased obviously (P<0.05) when injected with rutin at a dose of 20 and 50 microg g(-1) after 12 and 24 h, respectively. Both the THC and SOD activities of experimental and control groups revealed no significant difference at all doses. L. vannamei injected with rutin at either dose maintained lower glucose, lactate, and lipid levels in response to V. alginolyticus challenge after 12-36, 24-48, and 24-60 h, respectively. The survival rate of L. vannamei that received rutin at either dose was significantly higher than that received saline after 48-72 h. It was, therefore, concluded that the immune ability and resistance against V. alginolyticus infection of L. vannamei receiving rutin at > or = 10 microg g(-1) increased.

Ferritin administration effectively enhances immunity, physiological responses, and survival of Pacific white shrimp (Litopenaeus vannamei) challenged with white spot syndrome virus

We examined the physiological (hemolymph glucose, lactate, and lipid) and innate non-specific immune responses (total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (release of superoxide anion, O(2)(-)) and superoxide dismutase (SOD) activity) to white spot syndrome virus (WSSV) in white shrimp (Litopenaeus vannamei) that were individually injected with 0.1, 0.5, and 1 ng g(-1) ferritin. Results showed that the THC, PO activity, and respiratory bursts of L. vannamei obviously increased (p < 0.05) 12 h after being injected with any dose of ferritin. However, the THC, PO activity, and respiratory bursts of L. vannamei that had received 0.5 and 1 ng g(-1) ferritin were significant higher than those of the other groups at 36-60, 60-72, and 36-60 h, respectively. SOD activities of L. vannamei 12 h after receiving 0.1, 0.5, and 1 ng g(-1) ferritin were significantly higher than those receiving saline. L. vannamei injected with ferritin at any dose maintained lower glucose, lactate, and lipid levels in response to WSSV challenge after 12-36, 24-48, and 36-60 h, respectively. The survival of shrimp that had received 0.5 and 1 ng g(-1) ferritin was significantly higher than that of shrimp that received saline and of control shrimp after 72 h. The ferritin messenger RNA transcripts of shrimp that had received 0.5 and 1 ng g(-1) ferritin were significantly higher than that of shrimp that received saline after 36 h. It was, therefore, concluded that the immune ability and resistance against WSSV infection increased in L. vannamei that had received > 0.5 ng g(-1) ferritin. Ferritin does play important roles in the innate immunity of the white shrimp. We observed higher SOD activities of L. vannamei that had received 0.1, 0.5, and 1 ng ferritin after 12 h than those that had received only saline (control), and the high SOD expression remained at the same levels even after 72 h of treatment.

Effects of the water extract of Gynura bicolor (Roxb. & Willd.) DC on physiological and immune responses to Vibrio alginolyticus infection in white shrimp (Litopenaeus vannamei)

Gynura bicolor (Roxb. & Willd.) DC is widely distributed in certain areas of Asia and is very popular in vegetarian cuisine in Taiwan. To investigate the regulatory roles of G. bicolor in various functions in crustaceans, we examined innate non-specific immune responses (including total hemocyte count (THC), phenoloxidase activity (PO), respiratory bursts (RBs), and superoxide dismutase (SOD) activity), physiological responses (including haemolymph glucose, lactate, and lipids), and gene expressions (including prophenoloxidase (proPO), lipopolysaccharide- and b-1,3-glucan-binding protein (LGBP), and peroxinectin (PE) mRNA transcripts) to the pathogen Vibrio alginolyticus in white shrimp (Litopenaeus vannamei) that were individually injected with the water extract from G. bicolor at 2, 4, and 8 μg g(-1). Results indicated that PO, RBs, SOD activity, proPO, LGBP, and PE mRNA transcripts of shrimps receiving the water extract of G. bicolor at 2, 4, and 8 μg g(-1) significantly increased after challenge with V. alginolyticus for 96 h. However, no significant difference in the THC was seen at any dose. L. vannamei injected with the water extract of G. bicolor at all doses respectively maintained lower glucose, lactate, and lipid levels in response to V. alginolyticus challenge at 12-36, 24-36, and 24-48 h. Survival rates at 24-72 h of L. vannamei that received G. bicolor at any dose was significantly higher than those of shrimp that received saline. It was concluded that the water extract of G. bicolor can maintain physiological homeostasis and enhance immunity against V. alginolyticus infection in L. vannamei.

Effects of Panax notoginseng on the Metastasis of Human Colorectal Cancer Cells

The goal of this study was to investigate the effect of the Panax notoginseng ethanol extract (PNEE) on the regulation of human colorectal cancer (CRC) metastasis. The migratory, invasive, and adhesive abilities and the expression of metastasis-associated regulatory molecules in cultured human CRC cells (HCT-116) treated with the PNEE were analyzed in this study. The migratory and invasive abilities of HCT-116 cells were reduced after PNEE treatment. The incubation of HCT-116 cells with the PNEE for 24 h decreased MMP-9 expression and increased E-cadherin expression compared with the control group. The adhesion reaction assay indicated that treatment with the PNEE led to significantly decreased HCT-116 adhesion to endothelial cells (EA.hy926 cells). The integrin-1 protein levels in HCT-116 cells were significantly decreased following treatment with the PNEE. Similarly, the protein levels of E-selectin and intercellular adhesion molecule-1 (ICAM-1) were significantly decreased by treatment of the EA.hy926 endothelial cells with PNEE. A scanning electron microscope (SEM) examination indicated that HCT-116 cells treated with LPS combined with the PNEE had a less flattened and retracted shape compared with LPS-treated cells, and this change in shape was found to be a phenomenon of extravasation invasion. The transepithelial electrical resistance (TEER) of the EA.hy926 endothelial cell monolayer increased after incubation with the PNEE for 24 h. A cell-cell permeability assay indicated that HCT-116 cells treated with the PNEE displayed significantly reduced levels of phosphorylated VE-cadherin (p-VE-cadherin). These results demonstrate the antimetastatic properties of the PNEE and show that the PNEE affects cells by inhibiting cell migration, invasion, and adhesion and regulating the expression of metastasis-associated signaling molecules.

Sedanolide induces autophagy through the PI3K, p53 and NF-κB signaling pathways in human liver cancer cells.

Sedanolide (SN), a phthalide-like compound from celery seed oil, possesses antioxidant effects. However, the effect of SN on cell death in human liver cancer cells has yet to be determined. In this study, cell viability determination, monodansylcadaverine (MDC) fluorescent staining and immunoblot analysis were performed to determine autophagy induction and autophagy-induced protein expression changes via molecular examination after human liver cancer (J5) cells were treated with SN. Our studies demonstrate that SN suppressed J5 cell viability by inducing autophagy. Phosphoinositide 3-kinase (PI3K)-I, mammalian target of rapamycin (mTOR) and Akt protein levels decreased, whereas PI3K-III, LC3-II and Beclin-1 protein levels increased following SN treatment in J5 cells. In addition, SN treatment upregulated nuclear p53 and damage-regulated autophagy modulator (DRAM) and downregulated cytosolic p53 and Tp53-induced glycolysis and apoptosis regulator (TIGAR) expression in J5 cells. Furthermore, the cytosolic phosphorylation of inhibitor of kappa B (IκB) and nuclear p65 and the DNA-binding activity of NF-κB increased after SN treatment. These results suggest that SN induces J5 cell autophagy by regulating PI3K, p53 and NF-κB autophagy-associated signaling pathways in J5 cells.

Antimelanogenic, Antioxidant and Antiproliferative Effects of Antrodia camphorata Fruiting Bodies on B16-F0 Melanoma Cells

Antrodia camphorata is a fungus that is endemic to Taiwan, and its fruiting body has been used as a folk medicine for the prevention or treatment of diverse diseases. The present study is aimed at investigating the antimelanogenesis and antioxidation effect of the ethanolic extract of Antrodia camphorata fruiting body (EE-AC), as well as its antiproliferation effects in B16-F0 melanoma cells. Regarding antimelanogenic effects, EE-AC had effective cupric ions reducing capacity and expressed more potent inhibitory effect than kojic acid on mushroom tyrosinase activity. Moreover, EE-AC significantly inhibited cellular tyrosinase activity and the melanin content in B16-F0 cells at 12.5 μg/mL concentration without cell toxicities. Regarding antioxidant effects, EE-AC exhibited potent DPPH radical- and SOD-like-scavenging activities. Regarding antiproliferative effects, EE-AC exhibited a selective cytotoxic effect and markedly inhibited the migration ability of B16-F0 cells. EE-AC increased the population of B16-F0 cells at sub-G1 phase of the cell cycle. EE-AC also caused the increase of early apoptotic cells and chromatin condensation, which indicated the apoptotic effects in B16-F0 cells. We demonstrated that EE-AC possessed antimelanogenic, antioxidant and anti-skin cancer actions. The results would contribute to the development and application of cosmetics, healthy food and pharmaceuticals.

Chemopreventive Potential of 2,3,5,4′-Tetrahydroxystilbene-2-O-β-D-glucoside on the Formation of Aberrant Crypt Foci in Azoxymethane-Induced Colorectal Cancer in Rats

2,3,5,4′-Tetrahydroxystilbene-2-O-β-D-glucoside (THSG) has been shown to have antioxidative and anti-inflammatory effects. Oxidative and inflammatory reactions are related to the development of colorectal carcinoma (CRC). In the present study, we characterized the preventive activities of THSG on colon carcinogenesis using the azoxymethane- (AOM-) mediated rat colon carcinogenesis model. F344 male rats were randomly divided into 5 groups (untreated and AOM model rats treated with or without THSG at 30, 150, or 250 mg/kg) after which the numbers of aberrant crypt foci (ACF) were assessed in the colon tissues of all rats. The expressions of nuclear factor-κB (NF-κB), cyclooxygenase-2 (COX-2), matrix metalloproteinase proteins (MMPs), and carcinoembryonic antigen (CEA) were measured as effective early predictors of CRC using western blot analysis. Treatment with THSG (150 or 250 mg/kg) induced a 50% reduction in total colonic ACF formation (P < 0.05). Furthermore, our results revealed a downregulation of CEA and NF-κB protein levels in the reduced number of ACF elicited by treatment with THSG, whereas levels of COX-2 and MMPs proteins were not changed. Collectively, THSG may be a promising natural lead compound or drug candidate for treating early phases of CRC.