IntroductionAvascular necrosis of femoral head (ANFH) is a progressive disease that often leads to hip joint dysfunction and even disability in young patients. Although the standard treatment, which is core decompression, has the advantage of minimal invasion, the efficacy is variable. Recent studies have shown that implantation of bone marrow containing osteogenic precursors into necrotic lesion of ANFH may be promising for the treatment of ANFH.MethodsA prospective, double-blinded, randomized controlled trial was conducted to examine the effect of bone-marrow buffy coat (BBC) grafting combined with core decompression for the treatment of ANFH. Forty-five patients (53 hips) with Ficat stage I to III ANFH were recruited. The hips were allocated to the control group (core decompression + autologous bone graft) or treatment group (core decompression + autologous bone graft with BBC). Both patients and assessors were blinded to the treatment options. The clinical symptoms and disease progression were assessed as the primary and secondary outcomes.ResultsAt the final follow-up (24 months), there was a significant relief in pain (P <0.05) and clinical joint symptoms as measured by the Lequesne index (P <0.05) and Western Ontario and McMaster Universities Arthritis Index (P <0.05) in the treatment group. In addition, 33.3% of the hips in the control group have deteriorated to the next stage after 24 months post-procedure, whereas only 8% in the treatment group had further deterioration (P <0.05). More importantly, the non-progression rates for stage I/II hips were 100% in the treatment group and 66.7% in the control group.ConclusionImplantation of the autologous BBC grafting combined with core decompression is effective to prevent further progression for the early stages of ANFH.Trial registrationClinicalTrials.gov identifier NCT01613612. Registered 13 December 2011.

Butyrate has been reported to promote the performance and growth of chickens. The specific roles and efficacy of different sources of butyrate remained unclear. Thus, the present study aimed to investigate and compare the effects of Clostridium butyricum (CB), sodium butyrate (SB), and butyric acid glycerides (tributyrin, BAG) on the reproductive performance, egg quality, intestinal health, and offspring performance of yellow-feathered breeder hens. A total of 300 Lingnan yellow-feathered breeder hens were assigned to five treatment groups: control (CL), 1×108CFU/kg CB (CBL), 1×109CFU/kg CB (CBH), 500mg/kg SB, and 300mg/kg BAG. Results showed that the laying performance and egg quality were increased by CBL, CBH, and BAG. Both CB treatments increased the hatchability of fertilized eggs. Maternal supplementation with both levels of CB significantly elevated the growth performance of offspring. Treatment with CBL, CBH, SB, and BAG all improved the oviduct-related variables and reduced the plasmal antioxidant variables. The CBH, CBL, and BAG treatments also improved the intestinal morphology to different degrees. Jejunal contents of IL-6 were decreased by CBH and BAG, while those of IL-4, IL-6, IL-1β, and IgY were decreased by SB. Transcripts of nutrient transporters in jejunal mucosa were also upregulated by CBH, CBL, and SB treatments and expression of Bcl-2-associated X protein was decreased by CBL, CBH, and BAG. In cecal contents, CBL increased the abundance of Firmicutes and Bacillus, while CBH decreased the abundance of Proteobacteria. Also, the co-occurrence networks of intestinal microbes were regulated by CBH and BAG. In conclusion, dietary inclusion of CB and BAG improved the reproductive parameters, egg quality, and intestinal morphology of breeders. CB also influenced the hatching performance of breeders and growth performance of the offspring, while SB improved the oviduct-related variables. These beneficial effects may result from the regulation of cytokines, nutrient transporters, apoptosis, and gut microbiota; high-level CB had more obvious impact. Further study is needed to explore and understand the correlation between the altered gut microbiota induced by butyrate and the performance, egg quality, intestinal health, and also offspring performance.

Background/Aims: There has been increasing recent attention on the antioxidative capacity of equol. This study tested the effect of equol on oxidative stress induced by lipopolysaccharide (LPS) and regulation of immunity in chicken macrophages. Methods: Chicken HD11 macrophages were challenged with LPS (100 ng/mL) alone or with LPS (100 ng/mL) and (±)equol (10, 20, 40, 80, 160 μmol/L) together for 24h. Evaluated responses included the contents of malondialdehyde (MDA) and reduced glutathione (GSH), activities of total superoxide dismutase (T-SOD) and inducible nitric oxide synthase (iNOS), transcript abundance of superoxide dismutase 2 (SOD2), catalase (CAT), glutathione transferase (GST), Toll-like receptor 4 (TLR4), tumor necrosis factor alpha (TNFα) and interleukin-1 beta (IL-1β), and contents of the cytokines TNFα, IL-1β, interleukin-2 (IL-2) and interferon beta (IFNβ). Results: Exposure to LPS induced oxidative stress as contents of MDA increased and GSH decreased in LPS-treated cells (P < 0.05) compared to those in control cells. Compared to LPS alone, co-treatment with equol (20 μmol/L, 40 μmol/L or 80 μmol/L) reduced contents of MDA and increased those of GSH (both P < 0.05). Activity of T-SOD increased (P < 0.05) in cells treated with the higher contentration of equol (80 μmol/L or 160 μmol/L), however, all concentrations (20 μmol/L to 160 μmol/L) increased activity of iNOS (P < 0.05). The highest concentration of equol (160 μmol/L) increased SOD2 and GST transcripts (P < 0.05). Equol treatment increased transcripts of TLR4, TNFα and IL-1β (P < 0.05). And there were similar changes in contents of IL-1β, IL-2, IFNβ and TNFα in the cells (P < 0.05). Conclusions: It concluded that equol can protect chicken HD11 macrophages from oxidative stress induced by LPS through reducing lipid peroxidation products and enhancing contents of antioxidants, and activities of relevant antioxidase enzymes; effects were also seen in gene expression related to the immune response and increased contents of cytokines. The optimal concentration of equol on antioxidation and immune enhancement in chicken macrophages was 40 μmol/L.

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To investigate the prevalence and assess the zoonotic transmission burden of Cryptosporidium species in domestic pigeons in Guangdong Province, Southern China, 244 fecal samples were collected from four pigeon breeding farms between June 2012 and March 2013. Cryptosporidium oocysts were purified by Sheather's sugar flotation technique and characterized by DNA sequencing of small subunit ribosomal RNA (SSU rRNA) gene. Cryptosporidium species were determined by comparison of sequences with corresponding Cryptosporidium sequences in GenBank and phylogenetic analysis using neighbor-joining (NJ) in MEGA5.2. The overall prevalence of Cryptosporidium infection in domestic pigeons in Guangdong Province was 0.82% (2/244). Two Cryptosporidium species, namely Cryptosporidium baileyi and Cryptosporidium meleagridis, were identified in Huizhou and Chaozhou farm, respectively. These findings confirmed the existence of C. meleagridis infection in domestic pigeons in China for the first time and provided base-line information for further studies to evaluate the public health risk from pigeon to human.

BackgroundEvidence suggests that cytoglobin (Cygb) may function as a tumor suppressor gene.MethodsWe immunohistochemically evaluated the expression of Cygb, phosphatidylinositol-3 kinase (PI-3K), phosphorylated (p)-Akt, Interleukin-6 (IL-6), tumor necrosis factor-α (TNFα) and vascular endothelial growth factor (VEGF) in 88 patients with 41 high-grade gliomas and 47 low-grade gliomas. Intratumoral microvessel density (IMD) was also determined and associated with clinicopathological factors.ResultsLow expression of Cygb was significantly associated with the higher histological grading and tumor recurrence. A significant negative correlation emerged between Cygb expression and PI3K, p-Akt, IL-6, TNFα or VEGF expression. Cygb expression was negatively correlated with IMD. There was a positive correlation between PI3K, p-Akt, IL-6, TNFα and VEGF expression with IMD.High histologic grade, tumor recurrence, decreased Cygb expression, increased PI3K expression, increased p-Akt expression and increased VEGF expression correlated with patients’ overall survival in univariate analysis. However, only histological grading and Cygb expression exhibited a relationship with survival of patients as independent prognostic factors of glioma by multivariate analysis.ConclusionsCygb loss may contribute to tumor recurrence and a worse prognosis in gliomas. Cygb may serve as an independent predictive factor for prognosis of glioma patients.

This experiment investigated the antioxidant effects of equol on oxidative stress induced by H2O2 in chicken intestinal epithelial cells (IEC). IEC, from Lingnan yellow broiler chick embryos at embryonic day 18, were cultured in Dulbecco's modified Eagle's medium/F12. Cells were pretreated with 0, 10, 100, or 500 nM equol for 24 h before exposure to 300 μM H2O2 during a further 24 h. Oxidative damage was assessed by photomicrographs of cells, measuring cell proliferation, malondialdehyde (MDA) content, and antioxidative capacity from cellular total superoxide dismutase (T-SOD) activity, as well as the relative expressions of Nrf2, Bcl-2, SOD-1, GSH-Px3, Claudin-1 Treatment with 300 μM H2O2 caused serious damage to cells, with fewer normal intestinal epithelial cells, revealed by photomicroscopy. Treatment with 300 μM H2O2 significantly decreased live cell numbers compared with controls and prior treatment with equol had no effect in offsetting this action of H2O2 (P > 0.05). Compared with the cells treated just with H2O2, pre-treatment with 10, 100 and 500 nM equol significantly enhanced T-SOD activity (P < 0.05), while 10 and 100 nM equol before H2O2 significantly enhanced T-SOD activity compared with the untreated controls (P < 0.05). In cells pre-treated with 100 nM equol, the relative abundance of Nrf2 transcripts increased from the controls (P < 0.05) but expressions of Bcl-2, GSH-Px3, or SOD-1 were unaffected (P > 0.05). Pre-treatment with 10 and 100 nM equol significantly increased the transcript abundance of Claudin-1 (P < 0.05). Equol is shown here to protect IECs from oxidative damage by promoting the expression of antioxidant genes, increasing the activities of antioxidant enzymes, and by enhancing antioxidant capacity; 100 nM equol appeared to be the most effective concentration.

Loss-of-function ethylene insensitive 2 (EIN2) mutations showed ethylene insensitivity in Arabidopsis, which indicated an essential role of EIN2 in ethylene signaling. However, the function of EIN2 in fruit ripening has not been investigated. To gain a better understanding of EIN2, the temporal regulation of LeEIN2 expression during tomato fruit development was analyzed. The expression of LeEIN2 was constant at different stages of fruit development, and was not regulated by ethylene. Moreover, LeEIN2-silenced tomato fruits were developed using a virus-induced gene silencing fruit system to study the role of LeEIN2 in tomato fruit ripening. Silenced fruits had a delay in fruit development and ripening, related to greatly descended expression of ethylene-related and ripening-related genes in comparison with those of control fruits. These results suggested LeEIN2 positively mediated ethylene signals during tomato development. In addition, there were fewer seeds and locules in the silenced fruit than those in the control fruit, like the phenotype of parthenocarpic tomato fruit. The content of auxin and the expression of auxin-regulated gene were declined in silenced fruit, which indicated that EIN2 might be important for crosstalk between ethylene and auxin hormones. doi: 10.1111/j. 1672-9072.2006.00366.x; available online at www.blackwell-synergy.com, www.jipb.netThe plant hormone ethylene has a profound impact on plant growth and development, including fruit ripening (Abeles et al. 1992). Most components of the ethylene signal-transduction pathway have been identified in Arabidopsis thaliana (Kieber 1997;Guo and Ecker 2004). Ethylene is perceived by a family of receptors, and then the signal is transduced to the constitutive triple response (CTR), a protein kinase as the next component of the signal transduction pathway. Ethylene insensitive 3 (EIN3), the downstream of CTR, ultimately leads to the regulation of gene expression (Guo and Ecker 2004). EIN2, which acts as the downstream of CTR1 and the upstream of EIN3, plays an essential role in ethylene signal transduction (Chen and Bleecker 1995;Shibuya et al. 2004). In addition, EIN2 is also a cross-talk point between multiple hormone signaling pathways (Alonso et al. 1999;Adams-Phillips et al. 2004).Ethylene has a great impact on climacteric fruit ripening by coordinating the expression of genes (Alexander and Grierson 2002). The tomato is a typical climacteric fruit whose ripening is regulated by ethylene. Never ripe (Nr) tomato mutants that contain a mutation in the ethylene-binding domain of the NR receptor produce non-ripening fruits (Wilkinson et al. 1995). The delaying of fruit ripening is also observed in tomato plants with reduced expression of the tomato EIN3, like genes (LeEIL1, LeEIL2, and LeEIL3) (Fu et al. 2005). In addition, the overexpression of LeEIL1 can partially restore ripening in the Nr tomato mutant (Chen et al. 2004). This suggests that all Silencing of LeEIN2 Alters Fruit Ripening 1479 components of ethylene signaling might play an...