Dietary glutathione supplementation enhances antioxidant activity and protects against lipopolysaccharide-induced acute hepatopancreatic injury and cell apoptosis in Chinese mitten crab, Eriocheir sinensis.

Eriocheir sinensis (E. sinensis) is an important aquaculture species in China. However, deteriorating water environments lead to oxidative stress in these crabs, which subsequently reduces their quality and yield. Glutathione (GSH) is an endogenous antioxidant that is used to mitigate oxidative stress. However, whether dietary GSH can enhance the resistance of E. sinensis to oxidative stress remains unclear. Herein, crabs were fed dietary GSH (the basal diet was supplemented with 0, 300, 600, 900, and 1200 mg/kg diet weight of GSH) for up to 3 weeks and, then, challenged with lipopolysaccharide (LPS; 400 µg/kg body weight). After 6 h, their hepatopancreas were sampled. Diet supplementation with 600 and 900 mg/kg diet weight GSH not only increased the content of GSH in the hepatopancreas, but also enhanced the activities and mRNA expressions of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and glutathione-S-transferase (GST) (P < 0.05), compared to that in control crabs challenged with LPS alone. Diet supplementation with 600 or 900 mg/kg GSH also significantly increased the enzyme activities of GSH reductase and γ-glutamyl cysteine synthetase (γ-GCS) in LPS-treated crabs. Haematoxylin-eosin (HE) staining, electron microscopy, and flow cytometry were used to examine the structure and subcellular structure of and apoptosis in the hepatopancreas. The histopathology and sub-microstructure analysis results also showed that diet supplementation with 600 or 900 mg/kg GSH significantly alleviated damage in crabs challenged with LPS and decreased reactive oxygen species (ROS) levels and cell apoptosis ratios in the hepatopancreas, compared to the LPS-treated crabs. To further understand the effect of dietary GSH on LPS-induced apoptosis, the activities and gene or protein expressions of apoptosis-related factors were evaluated. As a result, diet supplementation with 600 or 900 mg/kg GSH significantly decreased the activities of caspases-3, -8, and -9 and inhibited the relative expression of caspase-3 and -8 but increased the expression of B-cell lymphoma-2 (bcl-2) and B-cell lymphoma-2-associated X inhibitor (bax inhibitor) in crabs challenged with LPS. This treatment further significantly downregulated the relative protein levels of caspase-3, -8, -9 and Bax and upregulated those of Bcl-2 in crabs challenged with LPS. However, treatment with 1200 mg/kg GSH caused the opposite effects. Overall, our results reveal that appropriate diets supplemented with 600 or 900 mg/kg GSH could enhance the antioxidant capacity and anti-apoptotic mechanisms in crabs after LPS injection, thereby providing a theoretical basis for the application of dietary GSH in E. sinensis.

Dietary reduced glutathione supplementation can improve growth, antioxidant capacity, and immunity on Chinese mitten crab, Eriocheir sinensis.

Eriocheir sinensis is an important aquaculture species in China, and its yield and quality are threatened by oxidative stress caused by deteriorating water conditions. Reduced glutathione (GSH) is an endogenous antioxidant, but whether dietary GSH can increase the resistance of E. sinensis to environmental stress remains unclear. Therefore, in this study, crabs were fed with dietary GSH (0, 300, 600, 900, and 1200 mg/kg diet weight) for up to 10 weeks to determine the effects of different dietary GSH concentrations on growth, antioxidant capacity, and immunity of E. sinensis. The results showed that the weight gain rate and survival rate increased significantly as dietary GSH levels increased from 0 to 900 mg/kg, but decreased at 1200 mg/kg. Compared with the control group, the diet supplemented with 900 mg/kg GSH not only increased the concentration of GSH in the haemolymph and hepatopancreas, but also enhanced the activity of glutathione peroxidase (GSH-Px) (p < 0.05). Diets supplemented with 600 or 900 mg/kg GSH significantly increased the enzymes activities of superoxide dismutase (SOD), lysozyme (LZM), alkaline phosphatase, and acid phosphatase, and significantly decreased the content of malondialdehyde. To understand the changes in the activity of these enzymes further, the expression of related genes was detected. Diets supplemented with 600 or 900 mg/kg GSH significantly upregulated the genes expressions of cytosolic manganese SOD, mitochondrial manganese SOD, copper, zinc-SOD, GSH-Px, LZM, and prophenoloxidase activating factor, and significantly down regulated the expression of Toll-like receptor 1, Toll-like receptor 2, Dorsal, and the myeloid differentiation factor 88. However, a diet supplemented with 1200 mg/kg GSH decreased those positive indicators. Overall, our results demonstrated that an appropriate diet supplemented with 600 or 900 mg/kg GSH enhances antioxidant capacity and immunity, which will enhance the general health of E. sinensis.

Effects of dietary supplementation with oridonin on the growth performance, relative organ weight, lymphocyte proliferation, and cytokine concentration in broiler chickens.

BACKGROUND: The present study was conducted to investigate the effects of oridonin (ORI) on growth performance, relative organ weight, lymphocyte proliferation, phagocytic function of neutrophils, and cytokine concentration in broiler chickens. A total of 240 one-day-old Arbor Acres male broilers were randomly assigned to four treatments with six replicate pens of 10 broiler chickens per pen. Broiler chickens were fed diets based on four levels of dietary ORI (0, 50, 80 and 100 mg/kg) for a 42-d feeding trial. The experimental diets were fed in three phases: 1 to 14 d, 15 to 28 d and 29 to 42 d. RESULTS: The results indicated that ORI has no influence on the growth performance (P > 0.05). However, ORI increased the relative weights of spleen and bursa, the number of proliferation peripheral blood T and B lymphocytes, the phagocytic rate of neutrophils, as well as the Interleukin-2 (IL-2), Interleukin-4 (IL-4) and tumor necrosis factor-a (TNF-a) serum concentrations in serum in broilers at days 14, 28 and 42 (P < 0.05). CONCLUSIONS: In conclusion, ORI can enhance immune function and resistance to disease in broiler chickens by stimulating T and B lymphocyte formation, division, and proliferation, as well as the modulation of Th1/Th2 cytokine secretion profiles.

Effect of dietary oridonin supplementation on growth performance, gut health, and immune response of broilers infected with Salmonella pullorum.

BACKGROUND: The effects of dietary supplementation of oridonin (ORI) on growth performance, cecal microbiota, epithelium development and antioxidant and immune parameters of broilers infected with S. pullorum were studied. A total of 300 1-d-old male chicks were selected and divided into 5 trial groups (6 replicates of 10 chickens): 1) nonchallenge control chicks (CON), 2) chicks treated with Salmonella Challenged Control (SCC), 3) chicks treated with S. pullorum challenge and 50 mg/kg ORI (O1), 4) chicks treated with S. pullorum challenge and 80 mg/kg ORI (O2), and 5) chicks treated with S. pullorum challenge and 100 mg/kg ORI (O3). RESULTS: The results showed that S. pullorum had no effect on the feed intake (FI), body weight gain (BWG) or feed conversion ratio (FCR) of broilers compared with the values measured for the CON group (P > 0.05). However, compared with the characteristics of CON, S. pullorum showed effects on the counts of Salmonella and Lactobacillus at 7 d and at 14 d (P < 0.05), on jejunal development at 7 d (P < 0.05), and on jejunal immunoglobulin A (IgA) concentration at 7 d (P < 0.05). The addition of 100 mg/kg ORI had the greatest effect on the counts of Lactobacillus and Salmonella in cecal content (P < 0.05), malonaldehyde (MDA) content in the jejunum (P < 0.05), villi height of the small intestine, and IgA concentrations in the jejunum (P < 0.05). CONCLUSIONS: The results suggest that ORI can improve Salmonella-induced immune responses and protect intestinal health, not only through its immune inhibitory properties but also through its multi-protective effects on gut health.

[Clinicopathological and prognostic analysis of primary clear cell carcinoma of the liver].

OBJECTIVE: To investigate the clinicopathological features and prognostic factors of primary clear cell carcinoma of the liver (PCCCL). METHODS: The clinical data of 41 PCCCL patients who underwent hepatic resection for PCCCL from October 1998 to June 2012 in our department were retrospectively analyzed. There were 31 male and 10 female patients. The median age was 56 years (range, 25 to 80 years), and the diagnosis was confirmed by postoperative pathological examination. The data of 106 well or moderately differentiated non-clear cell hepatocellular carcinoma (HCC) patients and 86 poorly differentiated non-clear cell HCC patients who underwent hepatic resection in the same period in our hospital in the same period were compared. The χ(2) test or Fischer's exact test, as appropriate, was used to compare group frequencies. Survival analysis was estimated by Kaplan-Meier method. Cox proportional hazards model was used in multivariate analysis. RESULTS: The proportion of fibrous capsule formation in the PCCCL tumors (46%, 19/41) was significantly higher than that of the other two groups (P < 0.05), whereas the PCCCL group had a lower rate of intravascular tumor embolus (2/41) and vascular invasion (1/41) (P < 0.05). The median survival time of PCCCL group was 65 months, the 1-, 3-, 5-year survival rates for PCCCL patients were 90.2%, 67.4% and 42.0%, significantly better than that of poor differentiated NCCHCC group's (82.9%, 33.3%, 7.2%, P < 0.01). However, there were no statistic significant differences between PCCCL group and well or moderately differentiated NCCHCC group (84.7%, 55.7%, 34.4%, P > 0.05). Tumor capsule formation was an independent favorable prognostic factor. In contrast, preoperative serum α-fetoprotein (AFP) level and hepatitis B virus infection were independent unfavorable prognostic factors for PCCCL. CONCLUSIONS: PCCCL is a rare, low degree malignant pathological subtype of HCC. Surgical resection may achieve favorable prognosis and even long-term survival.

Recovery of methane and acetate during ex-situ biogas upgrading via novel dual-membrane aerated biofilm reactor.

Biological biogas upgrading has been well-proven to be a promising approach for renewable bioenergy recovery, but hydrogen (H2)-assisted ex-situ biogas upgrading is hindered by a large solubility discrepancy between H2 and carbon dioxide (CO2). This study established a new dual-membrane aerated biofilm reactor (dMBfR) to improve the upgrading efficiency. Results showed that dMBfR operated at 1.25 atm H2 partial pressure, 1.5 atm biogas partial pressure, and 1.0 d hydraulic retention time could significantly improve the efficiency. The maximum methane purity of 97.6%, acetate production rate of 34.5 mmol L-1d-1, and H2 and CO2 utilization ratios of 96.5% and 96.3% were achieved. Further analysis showed that the improved performances of biogas upgrading and acetate recovery were positively correlated with the total abundances of functional microorganisms. Taken together, these results suggest that the dMBfR, which facilitates the precise CO2 and H2 supply, is an ideal approach for efficient biological biogas upgrading.

[Surgical management of nodular hyperplasia tumor-like hepatic lesions: a study of 72 cases].

OBJECTIVE: To explore and improve the diagnosis and treatment of benign hyperplasia tumor-like hepatic lesion. METHODS: The clinical data of 72 patients who had undergone hepatic resection for benign non-cystic hepatic lesions between January 1987 and December 2010 were analyzed retrospectively. There were 46 male and 26 female patients. The median age was 49 years (ranging 15 to 72 years), and diagnosis were confirmed by postoperative pathological examination. Thirty-four cases had symptoms, such as abdominal discomfort in right upper quadrant, fever, fatigue. All the cases had undergone hepatic resection; totally 78 lesions were removed. The surgical procedure including hepatic lobectomy for 13 cases, hepatic segmentectomy for 19 cases and non-anatomy resection for 39 cases. RESULTS: The finally diagnosis included focal nodular hyperplasia in 47 cases, adenomatous hyperplasia in 3 cases, hepatic dysplastic nodule in 3 cases, inflammatory pseudotumor in 3 cases, hepatic granuloma in 4 cases, nodular cirrhosis in 3 cases, hepatitis nodule in 6 cases, nodular regenerative hyperplasia in 1 case, lymphoid hyperplasia in 1 case. The postoperative complication rate was low (19.4%, 14/72). The follow-up period was 6 to 96 months. There was no mortality caused by lesion. One patient developed recurrence after 3 years. CONCLUSIONS: It is suggested that symptomatic lesions, lesions when malignancy cannot be excluded, and lesions which have canceration tendency, just like adenomatous hyperplasia, dysplastic nodule, and nodular cirrhosis regenerative hyperplasia need surgical resection. Operation is not necessary for other nodular hyperplasia lesions if the diagnoses are identified.

Carbonyl cyanide 3-chlorophenylhydrazone induced the imbalance of mitochondrial homeostasis in the liver of Megalobrama amblycephala: A dynamic study.

Carbonylcyanide-3-chlorophenylhydrazone (CCCP) is a protonophore, which causes uncoupling of proton gradient in the inner mitochondrial membrane, thus inhibiting the rate of ATP synthesis. However, this information is manly derived from mammals, while its effects on the mitochondrial homeostasis of aquatic animals are largely unknown. In this study, the mitochondrial homeostasis of a carp fish Megalobrama amblycephala was investigated systematically in a time-course manner by using CCCP. Fish was injected intraperitoneally with CCCP (1.8 mg/kg per body weight) and DMSO (control), respectively. The results showed that CCCP treatment induced hepatic mitochondrial oxidative stress, as was evidenced by the significantly increased MDA and PC contents coupled with the decreased SOD and MnSOD activities. Meanwhile, mitochondrial fission was up-regulated remarkably characterized by the increased transcriptions of Drp-1, Fis-1 and Mff. However, the opposite was true for mitochondrial fusion, as was indicative of the decreased transcriptions of Mfn-1, Mfn-2 and Opa-1. This consequently triggered mitophagy, as was supported by the accumulated mitochondrial autophagosomes and the increased protein levels of PINK1, Parkin, LC3-II and P62 accompanied by the increased LC3-II/LC3-I ratio. Mitochondrial biogenesis and function both decreased significantly addressed by the decreased activities of CS, SDH and complex I, IV and V, as well as the protein levels of PGC-1ß coupled with the decreased transcriptions of TFAM, COX-1, COX-2 and ATP-6. Unlikely, DMSO treatment exerted little influence. Overall, CCCP treatment resulted in the imbalance of mitochondrial homeostasis in Megalobrama amblycephala by promoting mitochondrial oxidative stress, fission and mitophagy, but depressing mitochondrial fusion, biogenesis and function.

Effects of oridonin on immune cells, Th1/Th2 balance and the expression of BLys in the spleens of broiler chickens challenged with Salmonella pullorum.

The aim of this study was to investigate the effects of oridonin (ORI) on the immune cells, Th1/Th2 balance and the expression of B lymphocyte stimulator (BLys) in the spleens of broilers infected with Salmonella pullorum. In a completely randomized design, 300 one-day-old AA male broilers were divided to 5 treatments. The groups included a noninfection control (CON) group receiveed a basal diet; a S. pullorum infect control group received the basal diet; and S. pullorum infect group receiveed the basal diet plus 50, 80, and 100 mg/kg ORI, respectively. The results showed that Salmonella challenge increased the relative weights of the spleen, white blood cell counts, lymphocyte and heterophil percentage, H/L ratio, the concentration and mRNA levels of spleen proinflammatory mediators, including tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-4, IL-6, and IL-10, as well as the anti-inflammatory target Blys (P < .05), and modulated the Th1/Th2 balance (P < .05). ORI pretreatment decreased the relative weight of the spleen and inhibited the release and expression of these proinflammatory mediators and the anti-inflammatory target BLys. The results suggested that ORI supplementation may have immunosuppressive and multiple modulation effects on activated microglia through modulation of the Th1/Th2 balance and BLys expression.