Identification of Driver Genes in Primary Liver Cancer by Integrating NGS and TCGA Mutation Data

Background: This study is aimed towards an exploration of mutant genes in primary liver cancer (PLC) patients by using bioinformatics and data mining techniques. Methods: Peripheral blood or paraffin-embedded tissues from 8 patients with PLC were analyzed using a 551 cancer-related gene panel on an Illumina NextSeq500 Sequencer (Illumina). Meanwhile, the data of 396 PLC cases were downloaded from The Cancer Genome Atlas (TCGA) database. The common mutated genes were obtained after integrating the mutation information of the above two cohorts, followed by functional enrichment and protein-protein interaction (PPI) analyses. Three well-known databases, including Vogelstein’s list, the Network of Cancer Gene (NCG), and the Catalog of Somatic Mutations in Cancer (COSMIC) database were used to screen driver genes. Furthermore, the Chi-square and logistic analysis were performed to analyze the correlation between the driver genes and clinicopathological characteristics, and Kaplan-Meier (KM) method and multivariate Cox analysis were conducted to evaluate the overall survival outcome. Results: In total, 84 mutation genes were obtained after 8 PLC patients undergoing gene mutation detection with next-generation sequencing (NGS). The top 100 most mutate gene data from PLC patients in TCGA database were downloaded. After integrating the above two cohorts, 17 common mutated genes were identified. Next, 11 driver genes were screened out by analyzing the intersection of the 17 mutation genes and the genes in the three well-known databases. Among them, RB1, TP53, and KRAS gene mutations were connected with clinicopathological characteristics, while all the 11 gene mutations had no relationship with overall survival. Conclusion: This study investigated the mutant genes with significant clinical implications in PLC patients, which may improve the knowledge of gene mutations in PLC molecular pathogenesis.

Leptin effects on the hypothalamic-pituitary-gonadal axis by activating proopiomelanocortin neurons Correlation between regulatory effects and duration

BACKGROUND：Leptin preserves reproductive functions by stimulating hypothalamic-pituitary-gonadal axis activities at different levels.Some interneurons play an important role in leptin regulation of the gonadal axis.It remains uncertain whether leptin regulates reproductive functions by activating proopiomelanocortin （POMC） neurons.OBJECTIVE：To investigate leptin effects on secretory function of the neuroendocrine-gonadal axis by activating POMC neurons and to observe and verify the relationship between leptin effects and various time points.DESIGN,TIME AND SETTING：The randomized,controlled,animal study was performed at the Basic Institute of Chengde Medical University and the Research Room of Reproductive Immunology of National Research Institute for Family Planning from June to September 2008.MATERIALS：Leptin （Peprotech,USA）,a-melanocyte stimulating hormone and rabbit anti-POMC polyclonal antibody （SC-20148） （Santa Cruz Biotechnology,USA）,follicle stimulating hormone,luteinizing hormone,and gonadotropin releasing hormone enzyme-labeled immunosorbent assay （ELISA） kit （ADL,USA） were used in the present study.METHODS：A total of 60 healthy,female,adult,Wistar rats received 17 （3-estradiol for 5 consecutive days at 15 days after ovariectomy.The rats were randomly assigned to physiological saline （n= 35）,leptin （n = 35）,and a-melanocyte stimulating hormone （n = 20） groups.MAIN OUTCOME MEASURES：Changes in gonadotropin releasing hormone,luteinizing hormone,and follicle stimulating hormone were compared following intraventricular injection of physiological saline,leptin,and a-melanocyte stimulating hormone at various time points.Changes in POMC mRNA and protein expression in the hypothalamus were measured following physiological saline and leptin injection via the lateral ventricle.RESULTS：Compared to the physiological saline group,leptin and a-melanocyte stimulating hormone affected secretion in the hypothalamus-pituitary axis.Leptin affected secretion of gonadotropin releasing hormone,luteinizing hormone,and follicle stimulating hormone,whereas a-melanocyte stimulating hormone inhibited secretion of these hormones.Compared to the physiological saline group,POMC mRNA expression was significantly increased in the hypothalamus at 2 and 4 hours after leptin injection （P〈 0.05）,but expression recovered to physiological saline group levels at 6 hours after injection （P 〉 0.05）.POMC protein expression was significantly increased in the hypothalamus at 4 and 6 hours after leptin injection （P〈 0.05）.CONCLUSION：Leptin affects secretory function of the neuroendocrine-gonadal axis through combined effects on POMC neurons and other pathways.Results suggested that the regulatory effects of POMC neurons were later compared to other neurons.

Effect of peroxiredoxin 6 on total and progressive motility of human spermatozoa after cryopreservation

Sperm cryopreservation is useful in assisted reproductive technology and male fertility preservation.However,freezing and thawing significantly reduces the total and progressive motility of human spermatozoa.In the present study,we explored the effects of peroxiredoxin 6(PRDX6)on total and progressive motility of human spermatozoa after cryopreservation.Semen samples of 20 males with normal parameters were collected and frozen in media supplemented with different concentrations of PRDX6(0 mM,10−5 mM,10−7 mM,and 10−9 mM,respectively).Postthaw total and progressive motility of sperms were measured.The results showed that in comparison with 0 mM,the concentrations of 10−5 mM,10−7 mM,and 10−9 mM of PRDX6 all significantly improved total motility and progressive motility of sperms(p<0.05).The 10−7 mM of PRDX6 showed the best performance.In conclusion,the supplementation of PRDX6 helps to maintain the total and progressive motility of human spermatozoa.