Effects of exogenous melatonin and photoperiod on sexual maturation in pullets.

Hy-Line Gray commercial pullets were maintained under 8-h photoperiods, 16-h photoperiods and 16-h photoperiods supplemented with a diet containing 20 or 200 mg/kg melatonin (MEL) to investigate the role of MEL in sexual development. A total of 256 Hy-Line Gray commercial pullets were placed, four birds to a cage, in four similar light-proof rooms (8-h photoperiod) at 6 weeks of age. At 70 day, three rooms containing a total of 192 birds were transferred to a 16-h photoperiod, whereas 64 birds were maintained under the 8-h photoperiod. Diets containing MEL at 20 and 200 mg/kg were fed to birds in two of the rooms under 16-h photoperiods. Birds maintained under an 8-h photoperiod matured 11.25 day later than those maintained under a 16-h photoperiod (p < 0.05). The group of birds receiving 20 mg/kg MEL matured 1.19 day later than those maintained under the 16-h photoperiod and 10.06 day earlier than those maintained under the 8-h photoperiod. The group of birds receiving 200 mg/kg MEL matured 3.13 day later than those maintained under a 16-h photoperiod and 8.12 day earlier than those maintained under an 8-h photoperiod. The average body weight of birds maintained under the 8-h photoperiod was greater than that of birds maintained under the 16-h photoperiod (p < 0.05) and was similar between the different MEL groups. The abdominal fat weight was lower in 16L:8D group compared with 8L:16D group (p < 0.05). The concentrations of follicle-stimulating hormone, luteinizing hormone, oestrogen and insulin did not differ significantly among the groups. The melatonin concentration in 200 mg/kg melatonin group was higher than that observed in the other groups; however, this concentration did not differ significantly (p > 0.05). These data suggest that the birds did not perceive the final 8-h photoperiod as being part of the night when they were given the MEL diets; continuously high plasma MEL was not observed in birds that responded as if they were in constant darkness. However, the later maturity of the groups administered MEL diets compared with the groups maintained under a constant 16-h photoperiod clearly indicated that MEL has some influence on the sexual maturity of pullets.

Molecular characterization of tumor necrosis factor receptor-associated factor 6 (TRAF6) in pearl oyster Pinctada martensii.

Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a key signaling adaptor molecule for tumor necrosis factor receptor superfamily and Toll-like receptor/interleukin-1 receptor family members. It signals the upstream receptors and is involved in a wide range of biological functions, such as immunity and bone metabolism. In this report, the TRAF6 gene from the pearl oyster Pinctada martensii (designated as PmTRAF6) was identified and characterized. The obtained full-length PmTRAF6 cDNA was 2273 bp, containing a 5'-untranslated region (UTR) of 297 bp, a 3'-UTR of 128 bp with a 42-bp poly (A) tail, and an open reading frame of 1848 bp that encoded 616-amino acid residues. The deduced protein sequence of PmTRAF6 contained a conserved TRAF family motif including a RING-type zinc finger, two TRAF-type zinc fingers, and a coiled-coil region followed by one meprin and TRAF homology domain. Multiple-sequence alignment indicated that TRAF6 was highly conserved among species, and PmTRAF6 showed 53% sequence identity to Azumapecten farreri and Mizuhopecten yessoensis. Furthermore, an amino acid sequence containing a low-complexity region was inserted in the TRAF6s from mollusk. Quantitative real-time polymerase chain reaction analysis demonstrated that PmTRAF6 was constitutively expressed in all tissues studied, with the most abundant mRNA expression in hepatopancreas and gill in P. martensii. After lipopolysaccharide stimulation, the expression of PmTRAF6 mRNA was dramatically upregulated. These results suggested that the obtained PmTRAF6 was a member of the TRAF6 family and perhaps involved in the innate immune response of pearl oyster.

Effect of dietary arginine and N-carbamoylglutamate supplementation on reproduction and gene expression of eNOS, VEGFA and PlGF1 in placenta in late pregnancy of sows.

The objectives of this study were to investigate the potential mechanisms of dietary arginine (Arg) and N-carbamoylglutamate (NCG) supplementation on reproductive performance of sows. Twenty-seven crossbred (Landrace×Large White) sows with similar body weight and parity at day (90±1) of gestation were assigned randomly into 3 groups (n=9) control group, Arg group, NCG group, and fed with the following diets: a control diet, and the control diet supplemented with 1.0% Arg or 0.1% NCG. Litter size was recorded. Blood samples were obtained for biochemical analyses. Placenta chorioallantoic membrane tissue collected immediately after birth to preserve in RNA stabilizer for mRNA analysis of endothelial nitric oxide synthase (eNOS), endothelial growth factor a (VEGFA) and placenta growth factor 1 (PlGF1) by real time-PCR. The results showed that compared with the control group, the average birth weight of all piglets born alive were 16.2% and 14.3% higher in the Arg and NCG groups (P<0.05), respectively; plasma VEGFA was higher in the Arg group (P<0.05). The expression of VEGFA in the allantochorion tissue of the NCG-supplemented group was higher (P<0.01), and tended to be higher in the Arg-supplemented group (0.05

Dietary supplementation with Astragalus polysaccharide enhances ileal digestibilities and serum concentrations of amino acids in early weaned piglets.

Two experiments were conducted to evaluate the effects of dietary supplementation with Astragalus polysaccharide (APS) on growth performance, apparent ileal digestibilities (AID) of amino acids (AA), and their serum concentrations in early weaned piglets. In Exp. 1, 60 pigs were weaned at 21 days of age (BW 7.35 +/- 0.23 kg) and allocated to three treatments (20 pigs/treatment), representing supplementing 0.0% (control), 0.02% colistin (antibiotic), or 0.1% APS to a corn- and soybean meal-based diet. Average daily gain (ADG), average daily feed intake (ADFI), and feed/gain ratio (F/G) were measured weekly. Blood samples were obtained from five pigs selected randomly from each treatment for the measurement of serum free AA concentrations on days 7, 14, and 28. In Exp. 2, 12 pigs were weaned at 21 day of age (BW 7.64 +/- 0.71 kg), assigned to three treatment groups as in Exp. 1, and surgically fitted with a simple T-cannula at the terminal ileum. Ileal digesta samples were obtained for the measurement of AID of AA on days 7, 14 and 28. Dietary APS did not affect ADFI, but enhanced (P < 0.05) ADG by 11 and 4.4%, and improved F/G by 5.6 and 8.4%, respectively, compared with the control and antibiotic groups. Addition of APS to the diet increased AID and serum concentrations of most nutritionally essential and non-essential AA (including arginine, proline, glutamate, lysine, methionine, tryptophan, and threonine) on days 14 and 28. Circulating levels of total AA were affected by the age of pigs and treatment x time interaction. Collectively, these findings indicate that APS may ameliorate the digestive and absorptive function and regulate AA metabolism to beneficially increase the entry of dietary AA into the systemic circulation, which provide a mechanism to explain the growth-promoting effect of APS in early weaned piglets.

Dietary supplementation with Chinese herbal powder enhances ileal digestibilities and serum concentrations of amino acids in young pigs.

This study was designed to determine the effect of ultra-fine Chinese herbal powder as a dietary additive on serum concentrations and apparent ileal digestibilities (AID) of amino acids (AA) in young pigs. In Experiment 1, 60 Duroc x Landrace x Yorkshire piglets weaned at 21 days of age were randomly assigned to one of three treatments, representing supplementation with 0 or 2 g/kg of the powder, or 0.2 g/kg of colistin (an antibiotic) to corn- and soybean meal-based diets (n = 20 per group). Blood samples from five piglets per group were collected on days 7, 14, and 28 to determine serum AA concentrations. In Experiment 2, 12 barrows with an average initial body weight of 7.64 kg were randomly assigned to one of the three dietary treatments, followed by surgical placement of a simple T-cannula at the terminal ileum. All of the diets contained 0.1% titanium oxide as a digestibility marker. The samples of terminal ileal digesta were collected on day 7 for determining AID of AA. Results show that dietary supplementation with the herbal powder increased (P < 0.05) serum concentrations and AID of most AA by 10-50% and 10-16%, respectively. As an indicator of improved intestinal function, AID values of calcium were also enhanced in piglets supplemented with the herbal powder. Dietary supplementation of colistin increased serum concentrations and AID values of some AA by 8-44% and 10-15%, respectively, in comparison with the non-supplemented group. These novel findings demonstrate that the herbal powder can enhance the digestibility of dietary protein and the intestinal absorption of AA into the systemic circulation in post-weaning pigs, therefore providing a new mechanism for its growth- and immunity-promoting efficacy.

Effect of dietary oligochitosan supplementation on ileal digestibility of nutrients and performance in broilers.

The effect of dietary chitosan oligosaccharides (COS) supplementation on ileal digestibilities of nutrients and performance in broilers was assessed by feeding graded levels (0, 50, 100, 150 mg/kg) of COS. Two thousand four hundred male commercial Avian broilers (1-d-old) were assigned randomly to 5 dietary treatment groups (60 birds per pen with 8 pens per treatment). Diet A was a typical corn- and soybean meal-based diet supplemented with 6 mg/kg of an antibiotic flavomycin (positive control). Diet B was the basal diet without any supplement. Diets C, D, and E were formulated by adding 50, 100, and 150 mg/kg of COS to the basal diet, respectively. On the morning of d 21 and 42, 64 birds (8 per pen with 8 pens per treatment) from the growth trial for each age group were killed by cervical dislocation for determination of the ileal digestibilities of nutrients. Dietary supplementation with COS and antibiotic enhanced (P < 0.05) the ileal digestibilities of DM, Ca, P, CP, and all amino acids (except for alanine in the 21-d-old birds or phenylalanine, glutamate, and glycine for the 42-d-old birds). Feed efficiency was improved (P < 0.05) in response to dietary supplementation of an antibiotic or COS (150 mg/kg for d 1 to 21, and 100 and 150 mg/kg for d 21 to 42). The results demonstrate for the first time to our knowledge that dietary COS supplementation was effective in increasing the ileal digestibilities of nutrients and feed efficiency in broilers. Our findings may explain a beneficial effect of COS on chicken growth performance.

Three xanthones and a benzophenone from Garcinia mangostana.

Investigation of the constituents of Garcinia mangostana has led to the isolation of four new compounds: three minor xanthones, garcimangosone A (1), garcimangosone B (2), and garcimangosone C (3), and a benzophenone glucoside, garcimangosone D (4). The structures of these four compounds were established by spectral (NMR and MS) and chemical methods.

Anti-hepatitis B virus effects of wogonin isolated from Scutellaria baicalensis.

By using an HBV-producing cell line (MS-G2) in vitro culture system, we found that wogonin isolated from Scutellaria baicalensis can suppress HBV surface antigen production (P < 0.001) without evidence of cytotoxicity. By assaying the endogenous HBV DNA polymerase activity, we found that both the relaxed circular and the linear forms of HBV DNA are significantly reduced in the wogonin-treated group. Wogonin deserves to be further evaluated for the treatment of human HBV infection.

Dihydrophenanthrenes from Spiranthes sinensis.

Six novel dihydrophenanthrene derivatives, sinensols A-F (1-6), were isolated from the aerial parts of Spiranthesis sinensis. Their structures were determined on the basis of various spectroscopic data, in particular those yielded by MS and 2D NMR techniques.

A new anti-HBeAg lignan, kadsumarin A, from Kadsura matsudai and Schizandra arisanensis.

A new C18 dibenzocyclooctadiene lignan, kadsumarin A (1) was isolated from Kadsura matsudai Hayata and Schizandra arisanensis Hayata. The anti-HBeAg test revealed that kadsumarin A had activity at a concentration of 40 micrograms/ml (= 90.1 microM). Its structural elucidation by spectral analysis was discussed in this note.

Anti-tumor effects of d-dicentrine from the root of Lindera megaphylla.

d-Dicentrine, a naturally occurring aporphine type isoquinoline alkaloid, isolated from the root of Lindera megaphylla Hemsl. (Lauraceae), was evaluated for its potential anti-cancer activity. We found d-dicentrine significantly inhibited the growth of human hepatoma cell line HuH-7 by delaying its doubling time in tissue culture. An in vitro colony forming assay showed that d-dicentrine decreased the colony formation efficiency in both hepatoma cell lines, HuH-7 and MS-G2, used in our study. Biosyntheses of the macromolecules DNA and RNA were also strongly inhibited. An MTT assay in 21 tumor cell lines also revealed that d-dicentrine was most cytotoxic to esophageal carcinoma HCE-6, lymphoma cell lines Molt-4 and CESS, leukemia cell lines HL60 and K562, and hepatoma cell line MS-G2. An in vitro tumor growing assay in the Severe Combined immunodeficiency (SCID) mice showed that intraperitoneal injection of d-dicentrine at the dose of 100 micrograms twice a week for 4 weeks significantly inhibited the tumor incidence of leukemia cell line K562 in SCID mice. All these data indicated that d-dicentrine has potential anti-tumor applications.

Osthole increases glycosylation of hepatitis B surface antigen and suppresses the secretion of hepatitis B virus in vitro.

During screening of Chinese herbal medicines for the activities against hepatitis B virus (HBV), a known pure compound, osthole, was found to inhibit the secretion of HBV surface antigens in vitro. The secretion of hepatitis B surface antigen (HBsAg) in culture medium of MS-G2 and HuH-7 cells transfected with HBV DNA decreased by 60% to 70% after osthole treatment, without any detectable cytotoxic effects. Both 42-nm Dane particles and 22-nm subviral HBsAg particles were significantly reduced as revealed by density gradient analysis of viral particles and the endogenous; HBV DNA polymerase assay. The production of naked HBcAg particles was not changed by osthole treatment. The steady-state level of HBV-specific messenger RNA (mRNA) was not altered by osthole treatment, as revealed by Northern blot analysis, Immunoprecipitation of culture medium showed that the ratio of glycosylated versus nonglycosylated HBsAg increased after osthole treatment. Kinetic studies revealed that osthole increased the glycosylation rate of HBsAg, thus altering the ratio of glycosylated to unglycosylated HBsAg. The increase of glycosylation of HBsAg by osthole was blocked by tunicamycin, an inhibitor of protein glycosylation. We conclude that osthole suppresses the secretion of HBV in cell culture probably because of its novel function in increasing the glycosylation of HBsAg.

[A report of fertilizer-spreading experiment of Scutellaria baicalensis Georgi].

The result shows that all the three kinds of fertilizers nitrogen, phosphorus and potassium can markedly raise the production of Scutellaria baicalensis. Both single spreading and mixed spreading help to enhance the yield, but mixed spreading gives higher yield--an increase of 26% over the non-spreading.