Effect of feeding linseed diet on testis development, antioxidant capacity, and epididymal cauda sperm concentration in Chinese Hu lamb.

Polyunsaturated fatty acids (PUFAs) are essential for mammalian testis development and sperm function. However, PUFAs that are contained in linseed oil are easily oxidized in the diet and biohydrogenated in the rumen. In this study, we investigated the effect of linseed as a source of PUFAs on the antioxidant capacity and testis development in Hu lamb. Seventy-five 3-month-old lambs were randomly assigned to three groups. Within each treatment group, 25 lambs were allocated to five pens (five lambs per pen). The lambs in the control group were fed a control diet without linseed for 42 days from D22 to D63. Group I (BS28) was fed a control diet from D22 to D35 and 8% linseed diet from D36 to D63. Group II (BS42) was fed an 8% linseed diet for 42 days from D22 to D63. After 63-day feeding trial, all lambs except the heaviest and lightest in each pen were humanely slaughtered and investigated. Results revealed that feeding linseed did not affect the body weight, scrotal circumference, and testis weight, whereas feeding linseed for 42 days increased the epididymis weight (37.85 ± 1.61 g vs. 32.09 ± 1.06 g, P < 0.05) compared with the control group. Feeding lambs with linseed for 42 days also significantly upregulated the expression of antioxidative (glutathione peroxidase 4 and copper-zinc superoxide dismutase), steroidogenesis (3ß-hydroxysteroid dehydrogenase and steroid acute regulatory protein), and PUFA metabolism-related genes (fatty acid desaturase 2 and elongation of very long-chain fatty acid protein 2) and proliferating cell nuclear antigen mRNA (P < 0.05). It also increased the relative expression of mitochondrial DNA (P < 0.05), total antioxidant capacity (0.230 ± 0.019 mmol/mgprot vs. 0.175 ± 0.011 mmol/mgprot, P < 0.05), and superoxide dismutase (1661.467 ± 147.117 U/mgprot vs. 1158.891 ± 98.850 U/mgprot, P < 0.05) in testicular tissue but decreased the cholesterol concentration (0.331 ± 0.073 mmol/mgprot vs. 0.671 ± 0.092 mmol/mgprot, P < 0.05) compared with the control group. Therefore, feeding lambs with linseed for 42 days stimulated seminiferous tubule development and increased the number of Sertoli cells (20.71 ± 0.89 vs. 17.6 ± 0.73, P < 0.05), epididymal cauda lumina diameter (638.26 ± 22.32 µm vs. 444.41 ± 34.80 µm, P < 0.05), and the number of sperm in the epididymal cauda (68.91 ± 7.06 × 108/g vs. 36.61 ± 7.50 × 108/g). All these results suggested that feeding linseed in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and PUFA metabolism-related genes; increased the antioxidant capacity in lamb's testis; and contributed to testis development and spermatogenesis.

Growth performance, rumen fermentation, bacteria composition, and gene expressions involved in intracellular pH regulation of rumen epithelium in finishing Hu lambs differing in residual feed intake phenotype.

The objective of this study was to evaluate the effect of residual feed intake (RFI) on rumen function in finishing lambs. A total of 60 male Hu lambs (average initial BW = 25.2 ± 2.5kg) were used and were offered a pelleted high-concentrate diet, of which the forage to concentrate ratio was 25:75. Individual feed intake was recorded over a period of 42 d, then 10 lambs with the lowest RFI and the highest RFI were selected, respectively. The rumen fluid used for fermentation variables and relative abundance of bacteria measurement was obtained on d 10 and 20 after RFI measurement. At the end of this experiment, the selected lambs were slaughtered and rumen epithelium and liver tissues were collected for RNA extraction. Low-RFI lambs had lower ( < 0.01) DMI and greater ( < 0.05) G:F than the high-RFI ones, while the RFI groups did not differ in ADG and BW ( > 0.05). Additionally, RFI was positively ( = 0.57; < 0.01) correlated with DMI and negatively ( = -0.53; < 0.05) correlated with G:F. Total VFA and individual VFA decreased ( < 0.05) over time. The concentrations of total VFA, acetate, valerate, isobutyrate, isovalerate, and rumen pH ( > 0.05) were not affected by RFI classification. Nonetheless, low-RFI group lambs had a greater ( < 0.05) concentration of propionate, a lower ( < 0.05) concentration of butyrate, and a lower ( < 0.05) acetate to propionate ratio compared with the high-RFI group. There was a significant ( < 0.05) effect of RFI on the relative abundance of and . The relative abundance of , , and decreased ( < 0.05) over time in high-RFI group. And the relative abundance of in high-RFI group was greater ( < 0.05) than its low-RFI counterpart. Furthermore, RFI had no effect ( > 0.05) on gene expression associated with intracellular pH regulation (, , , , , , , and ) in rumen epithelium and ß-hydroxybutyrate metabolism () in both rumen epithelium and liver tissues. In conclusion, even though low-RFI lambs had lower DMI, however, the number of was lower. Additionally, there was no difference in gene expressions level associated with intracellular pH regulation in rumen epithelium between RFI groups.

[Observation of the biological characterizations of nasopharyngeal epithelial cells by EB virus infection in early phase of immortalization].

The multi-stage cell model of the nasopharyngeal carcinoma development in vitro by Epstein-Barr virus transformation is beneficial for the elucidation of the mechanism of nasopharyngeal cancer. To observe the biological changes of primary human nasopharyngeal epithelial cells in early phase of immortalization, in this study, we have detected the morphological changes and the expression profile of senescence-associated beta-galactosidase (SA-beta-Gal) in primary culture. In addition, the expression of EB virus latent membrane protein 1 (LMP1) and the growth curve of primary cells were also detected. Our results showed a low percentage of cells infected with EB virus expressing SA-beta-Gal activity at the late primary culture. In morphology, the cells also formed multilayer foci, and the cell population doubling time was showed. These results demonstrated that the nasopharyngeal epithelial cells by EB virus infection have passed through the senescence and entered the early phase of immortalization. These cells have some of the transformed characteristics. Our results provided the data for further study on the mechanism of immortalization and the establishment of human nasopharyngeal epithelial cell line.

Nucleotide sequence analysis of a transforming gene isolated from nasopharyngeal carcinoma cell line CNE2: an aberrant human immunoglobulin kappa light chain which lacks variable region.

A transforming gene, designated Tx, was isolated from a human nasopharyngeal carcinoma (NPC) cell line CNE2 by transfection and molecular cloning techniques. The Tx gene was analyzed using computer-based bioinformatics and compared with the known sequences in EMBL and GenBank databases. We found that Tx contains human immunoglobulin kappa light chain constant region, five intact joining regions J1-J5, five recombination signal sequences and an N-segment besides classic regulatory sequences such as TATA boxes, CAAT boxes, poly A signals, etc. Interestingly, Tx also contains several binding sites for nuclear transcription factors such as NF-kappaB, NF-IL6, TFIID, etc. In conclusion, there are only several base pairs mutations or deletions compared with normal Ig K JC gDNA fragment. In all, Tx is an aberrant human immunoglobulin kappa light chain that contains the constant region, five joining regions, which lacks the variable regions.

[The effects of exogenous p16 expression on CDK4, Cyclin D1 and pRb in nasopharyngeal carcinoma cell lines].

OBJECTIVE: To investigate the effect of exogenous p16 on Cyclin D1, CDK4 and pRb, and to explore the mechanism of the growth suppression of p16 in nasopharyngeal carcinoma cell lines. METHODS: The curve of cell growth rate in three kinds of HNE1, # 3-2 and # 4-2 cell lines was analyzed and their double time was compared. Then the distribution of the cell cycle was detected by flow cytometry. The expression of p16 and the effect of exogenous p16 expression on CDK4, Cyclin D1 and pRb are studied by means of Western Blot. RESULTS: As compared with HNE1, # 3-2 and # 4-2 showed a longer double time(23.4 h vs 28.8 h, 31.2 h). # 4-2 showed a significant accumulation of cells in G0/G1 phase(P < 0.01) and decreasing in S phase(P < 0.05) while HNE1 and # 3-2 had no obvious difference(P > 0.05). Cyclin D1 expression was upregulated in # 4-2 but downregulated in # 3-2 by exogenous expressed p16. No obvious difference on CDK4 expression was found. Hypophosphorylated pRb was detected in three cell lines. The expression was stronger in # 4-2, and # 3-2 than that in HNE1 and Hela. Hyperphosphorylated pRb was also detected in HNE1. CONCLUSION: Exogenous p16 expression may arrest cell cycle in G0/G1 phase and suppress cell growth. The major mechanism is not to regulate the level of the expression of CDK4. There might be a threshold in p16 regulating Cyclin D1 expression. However, the final result contributes to the inhibition of pRb phosphorylation.