Transcriptomic analysis of the HPG axis-related tissues reveals potential candidate genes and regulatory pathways associated with testicular size in Hu sheep.

Testicular size is an excellent proxy for selecting high-fertility rams. The hypothalamus-pituitary-gonadal (HPG) axis plays an important role in regulating reproductive capacity in vertebrates, while key genes and regulatory pathways within the HPG axis associated with testicular size remain largely unknown in sheep. This study comprehensively compared the transcriptomic profiles in the hypothalamus, pituitary and testis of rams after sexual maturity between the large-testis group (LTG, testicular weight = 454.29 ± 54.24 g) and the small-testis group (STG, testicular weight = 77.29 ± 10.76 g). In total, 914, 795 and 10518 differentially expressed genes (DEGs) were identified in the hypothalamus, pituitary and testis between LTG and STG, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these DEGs were mainly involved in the biological processes of reproduction, biological regulation, and development process. Notably, the neuroactive ligand-receptor interaction and cAMP signaling pathways, commonly enriched by the DEGs in the hypothalamus and pituitary between two groups, were considered as two key signal pathways regulating testicular development through the HPGs axis. Weighted gene co-expression network analysis (WGCNA) identified two modules that were significantly associated with testicular size, and 97 key genes were selected with high module membership (MM) and gene significance (GS) in these two modules. Finally, a protein-protein interaction (PPI) network was constructed, and ten genes with the highest degree were represented as hub genes, including FOS, NPY, SST, F2, AGT, NTS, OXT, EDN1, VIP and TAC1. Taken together, these results provide new insights into the molecular mechanism underlying the HPG axis regulating testicular size of Hu sheep.

Dietary supplementation of grape seed tannin extract stimulated testis development, changed fatty acid profiles and increased testis antioxidant capacity in pre-puberty hu lambs.

Grape seed tannin extract (GPE) from wine grape pomace has many effective anti-oxidative effects and is used as a promising natural feed additive in the animal feed industry. This study investigated the effect of GPE as a source of tannin on the antioxidant capacity and testis development in Hu lambs. Twenty-seven 3-month-old ram lambs were randomly assigned to three groups. For each treatment group, nine lambs were allocated to nine pens (one lamb per pen). The lambs in the control group were fed a control diet without GPE for 61 days from D21 to D80. Group I (TAN1) was fed with 0.36% GPE diet, and Group II (TAN2) was fed with 0.72% GPE diet. After an 81-day feeding trial, all lambs except the heaviest and lightest in each group were humanely slaughtered and investigated. Results showed that feeding GPE did not affect the body weight, average daily gain, dry matter intake, scrotal circumference, and testis index. Meanwhile, feeding with 0.36% GPE diet increased testis weight, testis volume, and epididymis weight (P ≤ 0.05) compared with those of the control, but no difference was found between TAN1 and TAN2 groups. Copper-zinc superoxide dismutase (Cu-ZnSOD), steroid acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), follicle-stimulating hormone receptor (FSHR), elongation of very long chain fatty acid protein 2 (ELOVL2), fatty acid desaturase (FADS2), and proliferating cell nuclear antigen (PCNA) mRNA in TAN1 and TAN2 groups were significantly up-regulated (P < 0.05). GPE also markedly increased the antioxidant status of testis. Compared with the control group, the treatment groups showed significantly increased superoxide dismutase (SOD) activity (314.23 ± 18.64 U/mg prot in control, 505.22 ± 63.47 U/mg prot in TAN1 and 587.88 ± 55.94 U/mg prot in TAN2, P < 0.05) and total antioxidant capacity (T-AOC) (98.23 ± 18.99 U/g prot in control, 202.15 ± 34.19 U/g prot in TAN1 and 189.57 ± 18.95 U/g prot in TAN2, P < 0.05). Consuming 0.72% GPE also changed the fatty acid profiles in testis with increased C15:1, C22:6n3, and total n-3 fatty acids (P < 0.05) but decreased C22:5n3 (P < 0.05). Therefore, feeding lambs with GPE stimulated testis seminiferous tubule development and increased the number of Sertoli cells (10.56 ± 0.44 in control, 14.10 ± 0.57 in TAN1 and 13.60 ± 0.42, P < 0.05), and seminiferous tubule diameter (109.30 ± 4.56 µm in control, 164.49 ± 5.37 µm in TAN1 and 146.56 ± 4.53 µm in TAN2, P < 0.05). These results suggested that feeding GPE in the early reproductive development stage of lambs upregulated the expression of antioxidative, steroidogenesis, and polyunsaturated fatty acid metabolism-related genes, changed the fatty acid profiles, increased the antioxidant capacity in lamb's testis, and contributed to testis development and spermatogenesis.

[Stored autologous blood transfusion does not increase recurrence of prostate cancer after radical prostatectomy: A preliminary study].

OBJECTIVE: To analyze the association of stored autologous blood transfusion (SABT) with tumor recurrence in PCa patients after radical prostatectomy and explore the application of SABT in this surgical procedure. METHODS: Forty-five PCa patients underwent radical prostatectomy in our hospital in recent five years, of whom, 20 received SABT (group A) and the other 25 allogeneic blood transfusion (group B) intraoperatively. After surgery, we followed up the patients regularly for 3－66 months by examination of the levels of total PSA (tPSA) and free PSA (fPSA), digital rectal examination (DRE), and MRI to observe the biochemical recurrence of the tumor. We compared the data obtained between the two groups of patients. RESULTS: In group A, 8 cases were in stages T1a－T1b and 12 in stages T2a－T2c, and in group B, 14 cases were in stages T1a－T1b and 11 in stages T2a－T2c. The volume of transfused blood was 800 ml in group A and 400－1 200 ml in group B. No statistically significant differences were observed between the two groups in the operation time, intraoperative blood loss or postoperative Gleason scores (P > 0.05), nor in the tPSA level or the results of DRE and MRI at 12, 24, 36, 48 and over 48 months (P > 0.05). CONCLUSIONS: SABT is safe for PCa patients undergoing radical prostatectomy and does not increase the tumor recurrence rate after surgery.

Supplementation with dietary linseed oil during peri-puberty stimulates steroidogenesis and testis development in rams.

Omega-3 polyunsaturated fatty acids (ω-3 PUFAs), such as α-linolenic acid (ALA), eicosapentaenoic acid, and docosahexaenoic acid, are involved in male reproductive function. In this study, we investigated the effects of linseed oil (LO) as a source of ALA on the steroidogenesis and changes of testicular histology in rams. Sixteen 3-month old rams during peri-puberty were randomly assigned into two groups. Eight rams were assigned as the control group, and the other received LO (4% dry matter of total diet) as the LO treatment group. After an 81-day feeding trial, the rams were slaughtered and investigated. Results revealed that compared with control group, diet containing LO did not affect body weight (36.87 ± 0.53 kg vs. 37.65 ± 0.64 kg, respectively; P = 0.361), average daily gain (227.47 ± 5.82 g vs. 237.95 ± 9.22 g, respectively; P = 0.353) and epididymis weight (40.77 ± 4.41 g vs. 45.53 ± 4.01 g, respectively; P = 0.398), however, it up-regulated PUFAs metabolism and steroidogenesis-related genes mRNA expression (P < 0.05), and increased plasma estradiol concentration (14.88 ± 0.67 pg/mL vs. 19.50 ± 1.27 pg/mL, respectively; P < 0.05). Therefore, LO stimulated seminiferous tubule development and increased the number of Sertoli cells (19.17 ± 2.14 vs. 27.2 ± 2.39, respectively; P < 0.01), germ-cell layers, as well as testis weight (148.65 ± 22.66 g vs. 249.96 ± 30.63 g, respectively; P < 0.05). All these results suggested that LO can improve testis development during peri-puberty by regulating steroidogenesis in rams' testes.

[Studies on fingerprints of Centella asiatica by HPLC].

OBJECTIVE: To study the HPLC fingerprints and establish a sensitive and specific method for controlling the quality of Centella asiatica. METHODS: HPLC gradient elution was applied for the fingerprints of Centella asiatica. All 16 samples are collected from different habitats of China. The columni was Alltech C18 column (250 mm x 4.6 mm, 5 microm), mobile phase was acetonitrile-water, flow rate was 1.0 ml/min, wavelength was 205 nm. RESULTS: The fingerprint of Centella asiatica was established, 16 samples of different areas of Centella asiatica were detected. There were 15 common peaks in the HPLC fingerprints of Centella asiatica. By comparison with the reference standards and using LC-ESI-MS(n) to corroborate the structure, 5-10 peaks were identified as madecassoside, asiaticoside, quercetin, kaemperol, madecassic acid and asiatic acid respectively. After calculating the similarity of the HPLC fingerprints of 16 habitants, the similarity of different habitats has been bad quite. CONCLUSION: The method is accurate, reliable and good repeatability. This chromatographic fingerprint method can be used to controll the quality of Centella asiatica.