The dysfunction of hormone-sensitive lipase induces lipid deposition and reprogramming of nutrient metabolism in fish.

Hormone-sensitive lipase (HSL) is one of the rate-determining enzymes in the hydrolysis of TAG, playing a crucial role in lipid metabolism. However, the role of HSL-mediated lipolysis in systemic nutrient homoeostasis has not been intensively understood. Therefore, we used CRISPR/Cas9 technique and Hsl inhibitor (HSL-IN-1) to establish hsla-deficient (hsla-/-) and Hsl-inhibited zebrafish models, respectively. As a result, the hsla-/- zebrafish showed retarded growth and reduced oxygen consumption rate, accompanied with higher mRNA expression of the genes related to inflammation and apoptosis in liver and muscle. Furthermore, hsla-/- and HSL-IN-1-treated zebrafish both exhibited severe fat deposition, whereas their expressions of the genes related to lipolysis and fatty acid oxidation were markedly reduced. The TLC results also showed that the dysfunction of Hsl changed the whole-body lipid profile, including increasing the content of TG and decreasing the proportion of phospholipids. In addition, the systemic metabolic pattern was remodelled in hsla-/- and HSL-IN-1-treated zebrafish. The dysfunction of Hsl lowered the glycogen content in liver and muscle and enhanced the utilisation of glucose plus the expressions of glucose transporter and glycolysis genes. Besides, the whole-body protein content had significantly decreased in the hsla-/- and HSL-IN-1-treated zebrafish, accompanied with the lower activation of the mTOR pathway and enhanced protein and amino acid catabolism. Taken together, Hsl plays an essential role in energy homoeostasis, and its dysfunction would cause the disturbance of lipid catabolism but enhanced breakdown of glycogen and protein for energy compensation.

PDK inhibition promotes glucose utilization, reduces hepatic lipid deposition, and improves oxidative stress in largemouth bass (Micropterus salmoides) by increasing pyruvate oxidative phosphorylation.

In omnivorous fish, the pyruvate dehydrogenase kinases (PDKs)-pyruvate dehydrogenase E1α subunit (PDHE1α) axis is essential in the regulation of carbohydrate oxidative catabolism. Among the existing research, the role of the PDKs-PDHE1α axis in carnivorous fish with poor glucose utilization is unclear. In the present study, we determined the effects of PDK inhibition on the liver glycolipid metabolism of largemouth bass (Micropterus salmoides). DCA is a PDK-specific inhibitor that inhibits PDK by binding the allosteric sites. A total of 160 juvenile largemouth bass were randomly divided into two groups, with four replicates of 20 fish each, fed a control diet and a control diet supplemented with dichloroacetate (DCA) for 8 weeks. The present results showed that DCA supplementation significantly decreased the hepatosomatic index, triglycerides in liver and serum, and total liver lipids of largemouth bass compared with the control group. In addition, compared with the control group, DCA treatment significantly down-regulated gene expression associated with lipogenesis. Furthermore, DCA supplementation significantly decreased the mRNA expression of pdk3a and increased PDHE1α activity. In addition, DCA supplementation improved glucose oxidative catabolism and pyruvate oxidative phosphorylation (OXPHOS) in the liver, as evidenced by low pyruvate content in the liver and up-regulated expressions of glycolysis-related and TCA cycle/OXPHOS-related genes. Moreover, DCA consumption decreased hepatic malondialdehyde (MDA) content, enhanced the activities of superoxide dismutase (SOD), and increased transforming growth factor beta (tgf-ß), glutathione S-transferase (gst), and superoxide dismutase 1 (sod1) gene expression compared with the control diet. This study demonstrated that inhibition of PDKs by DCA promoted glucose utilization, reduced hepatic lipid deposition, and improved oxidative stress in largemouth bass by increasing pyruvate OXPHOS. Our findings contribute to the understanding of the underlying mechanism of the PDKs-PDHE1α axis in glucose metabolism and improve the utilization of dietary carbohydrates in farmed carnivorous fish.

Epigenetic Suppression of GADs Expression is Involved in Temporal Lobe Epilepsy and Pilocarpine-Induced Mice Epilepsy.

Recent studies have shown that histone acetylation is involved with the regulation of enzyme glutamate decarboxylases (GADs), including GAD67 and GAD65. Here, we investigated the histone acetylation modifications of GADs in the pathogenesis of epilepsy and explored the therapeutic effect of a novel second-generation histone deacetylase inhibitor (HDACi) JNJ-26481585 in epilepsy animals. We revealed the suppression of GADs protein and mRNA level, and histone hypoacetylation in patients with temporal lobe epilepsy and pilocarpine-induced epilepsy mice model. Double-immunofluorescence also indicated that the hypoacetyl-H3 was located in hippocampal GAD67/GAD65 positive neurons in epilepsy mice. JNJ-26481585 significantly reversed the decrease of the GAD67/GAD65 both protein and mRNA levels, and the histone hypoacetylation of GABAergic neurons in epilepsy mice. Meanwhile, single-cell real-time PCR performed in GFP-GAD67/GAD65 transgenic mice demonstrated that JNJ-26481585 induced increase of GAD67/GAD65 mRNA level in GABAergic neurons. Furthermore, JNJ-26481585 significantly alleviated the epileptic seizures in mice model. Together, our findings demonstrate inhibition of GADs gene via histone acetylation plays an important role in the pathgenesis of epilepsy, and suggest JNJ-26481585 as a promising therapeutic strategy for epilepsy.

A questionnaire study on attitudes toward birth and child-rearing of university students in Japan, China, and South Korea.

This study examines the attitudes of young Japanese, Chinese, and South Koreans toward birth and child-rearing. The survey targeted four-year university students (n＝1,668) who responded to an anonymous survey using self-report questionnaires between December 2012 and April 2013. The collection rates were 72.5%, 94.7%, and 96.5% for the Japanese, Chinese, and South Korean students, respectively. Correlations among the respondents' attributes, medical and scientific literacy levels, and views of preferred qualities of children were analyzed using chi-square test, supplemented by residual analysis (significance level set at p＜0.05). Participants were asked whether they were willing to use the following methods for obtaining preferred qualities in their children:(1) choosing a spouse (43.2%, 72.6%, and 85.1% of the Japanese, Chinese, and South Koreans, respectively, agreed);(2) using a sperm bank (cryobank) (5.8%, 60.1%, and 81.7% of the Japanese, Chines, and South Koreans, respectively, agreed);and (3) using an egg cell bank (ova bank or cryobank) (5.3%, 47.2%, and 70.3% of the Japanese, Chinese, and South Koreans, respectively, agreed). The proportion of affirmative responses (indicating "eugenic inclination") to these statements was significantly higher among the Chinese and South Korean participants than their Japanese counterparts (p＜0.001). Significant differences were also found in the attitudes of the 3 groups toward methods for obtaining the preferred qualities for their children:prenatal diagnosis, pre-implantation diagnosis, the environment during pregnancy, and child-rearing.

Dietary docosahexaenoic acid reduces fat deposition and alleviates liver damage induced by D-galactosamine and lipopolysaccharides in Nile tilapia (Oreochromis niloticus).

Liver health is important to maintain survival and growth of fish. Currently, the role of dietary docosahexaenoic acid (DHA) in improving fish liver health is largely unknown. This study investigated the role of DHA supplementation in fat deposition and liver damage caused by D-galactosamine (D-GalN) and lipopolysaccharides (LPS) in Nile tilapia (Oreochromis niloticus). Four diets were formulated as control diet (Con), Con supplemented with 1 % DHA, 2 % DHA and 4 % DHA diets, respectively. The diets were fed to 25 Nile tilapia (2.0 ± 0.1 g, average initial weight) in triplicates for four weeks. After the four weeks, 20 fish in each treatment were randomly selected and injected with a mixture of 500 mg D-GalN and 10 µL LPS per mL to induce acute liver injury. The results showed that the Nile tilapia fed on DHA diets decreased visceral somatic index, liver lipid content and serum and liver triglyceride concentrations than those fed on the Con diet. Moreover, after D-GalN/LPS injection, the fish fed on DHA diets decreased alanine aminotransferase and aspartate transaminase activities in the serum. The results of liver qPCR and transcriptomics assays together showed that the DHA diets feeding improved liver health by downregulating the expression of the genes related to toll-like receptor 4 (TLR4) signaling pathway, inflammation and apoptosis. This study indicates that DHA supplementation in Nile tilapia alleviates the liver damage caused by D-GalN/LPS through increasing lipid catabolism, decreasing lipogenesis, TLR4 signaling pathway, inflammation, and apoptosis. Our study provides novel knowledge on the role of DHA in improving liver health in cultured aquatic animals for sustainable aquaculture.

[Effects of bushen yinao tablet on physiology and cerebral gene expression in senescence-accelerated mice].

OBJECTIVE: To study the effects of Bushen Yin' ao Tablet (BSYNT) on physiology and cerebral gene expression in senescence-accelerated mice (SAM). METHODS: The change of cerebral tissues mRNA expression in SAM was analyzed and compared by messenger ribonucleic acids reverse transcription differential display polymerase chain reaction (mRNA DDRT-PCR) between the medicated group and the control group. RESULTS: BSYNT could increase the level of hemoglobin (Hb) and amount of erythrocyte (RBC) of blood deficiency mice, improve the spatial learning and memory function and the escape response by conditional stimulus. In this study, 14 differential display bands had been discerned, and three of them had been sequenced. The sequence of the three fragments was similar to fatty acid binding protein 7, ubiquinol-cytochrome C reductase complex (7. 2 kD) and 60S ribosomal protein L21 respectively. And the homogeneity was 97% , 100% , and 99% , respectively. CONCLUSION: BSYNT has effect on the physiological changing of mice, and its effect on cerebral tissues mRNA expression maybe play an important role in anti-aging on the molecular level.

[Detecting multi-drug resistance of bladder cancer for the intravesical chemotherapy].

OBJECTIVE: To explore multi-drug resistance (MDR) of bladder cancer for the intravesical instillation. METHODS: Using immunohistochemical staining, in 44-case human bladder cancer cells, the expressions of P-glycoprotein (P-gp), glutathione S-transferase (GST-pi) and topoisomerase (TOPO-II), were detected to find out the resistance to drugs. RESULTS: P-gp had a higher expression in 54.5% cases. GST-pi had no or a lower expression in 65.9% cases. TOPO-II had a higher expression in 29.5% but a lower expression in 65.9% cases. CONCLUSION: Detecting the factors of MDR in bladder cancer cells could help to choose drugs for intravesical chemotherapy.

[Expression of FANCG gene in acute myeloid leukemia].

This study was purposed to investigate the relationship between expression of the FANCG gene and adult sporadic acute myeloid leukemia (AML), real-time PCR with SYBR Green I technique was used for detecting FANCG gene expression level in bone marrow mononuclear cells of 54 newly diagnosed AML patients, 46 AML patients in complete remission (CR) and 36 control samples. ß-actin gene was used as internal reference. Relative changes of FANCG gene expression level were detected by 2(-ΔΔCT) method in newly diagnosed AML patients and control samples, in newly diagnosed AML and patient in CR, as well as in AML patients in CR and control samples. The results showed that the relative expression level of FANCG mRNA was 0.56 ± 0.27 in newly diagnosed group, 0.75 ± 0.54 in AML CR group, and 0.85 ± 0.45 in control group. The expression level of FANCG mRNA in newly diagnosed group was significantly lower than that in control and AML CR groups (P < 0.05). There was no statistically significant deference in comparison of AML CR group with the control group (P > 0.05). It is concluded that the expression of FANCG gene decrease in the newly diagnosed AML patients. There is no significant difference between AML CR group and control group, which indicated that FANCG gene may be related with the onset and the prognosis of AML, and may provide a clinical value for evaluating effect of chemotherapy.

Sonochemical synthesis of silver nanorods by reduction of silver nitrate in aqueous solution.

The sonochemical synthesis of silver nanorods has been achieved by ultrasonic irradiation of the aqueous solution of silver nitrate, methenamine (HMTA) and poly (vinyl pyrrolidone) (PVP) for 60 min. The silver nanorods obtained have lengths of 4-7 microm and mean diameters of about 100 nm. The structures of the samples were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), selected area electron diffraction (SAED) and X-ray powder diffraction (XRD), and the chemical composition of the sample was examined by energy-dispersive X-ray spectrum (EDS). The effects of the irradiation time, the concentration of PVP and the reaction temperature on the morphology of silver nanorods were discussed, and the mechanism of the silver nanorods formation was tentatively inferred.

Synthesis of dendritic silver nanostructures by means of ultrasonic irradiation.

In this paper, a simple and effective route for the synthesis of silver dendritic nanostructures by means of ultrasonic irradiation has been developed. Well-defined silver dendritic nanostructures were obtained by sonicating the aqueous solution of 0.04mol/L silver nitrate with 4.0mol/L isopropanol as reducing agent and 0.01mol/L PEG400 as disperser for 2h. The effects of the irradiation time, the concentration of Ag(+) and the molar ratio of PEG to AgNO(3) on the morphology of silver nanostructures were discussed. The structures of the obtained samples were characterized by transmission electron microscopy (TEM), selected area electron diffraction (SAED) and X-ray powder diffraction (XRD), and the chemical composition of the dendrites was examined by energy-dispersive X-ray spectrum (EDS).