Physiological Effects and Transcriptomic Analysis of sbGnRH on the Liver in Pompano (Trachinotus ovatus).

Pompano (Trachinotus ovatus) is one of the important economic marine fishes in the south coast of China. At present, the research on the basic biology of pompano is relatively weak, which has seriously affected the development of this economic important fish. The liver is an important digestive and metabolic organ of fish which plays an important regulatory role in its growth and development. It is necessary to clarify the effects of sea bream gonadotropin releasing hormone (sbGnRH) on liver physiology and metabolic enzyme activity. The effects of sbGnRH peptides (10 ng/gbw) on the physiological and biochemical indices and metabolic enzyme activities of pompano liver were studied. It was found that after injection of 10 ng/gbw sbGnRH peptides, the contents of albumin, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, glucose, creatine kinase, iron, magnesium, aspartate aminotransferase, alanine aminotransferase and creatinine increased, while of cholesterol and calcium contents decreased. The activities of amylase, lipase, pyruvate kinase, acyl CoA oxidase, superoxide dismutase, phospholipid hydroperoxide glutathione peroxidase, catalase, glucose-6-phosphate dehydrogenase, fatty acid synthase and lipoprotein lipase increased, while the activities of malic enzyme, carnitine acyl, carnitine translocation, acetyl CoA carboxylase and malondialdehyde decreased. Three hours after the injection of different concentrations of sbGnRH peptides (0 and 10 ng/gbw), the transcriptome sequences of the two groups of livers were sequenced. After quality control and removal of some low-quality data, clean reads of 21,283,647、19,427,359、21,873,990、21,732,174、23,660,062 and 21,592,338 were obtained respectively. In this study, 99 genes were screened and identified as differentially expressed genes, including 77 up-regulated genes and 22 down-regulated genes. According to the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathway analyses, these pathways and the typical genes involved can be divided into cellular processes, environmental information processing, genetic information processing, diseases, metabolism and organismal systems. The results from this study provide a the oretical basis for studying the effects of sbGnRH on the physiology, biochemistry and metabolic enzyme activities of liver in pompano.

Identification and functional characterization of gonadotropin -releasing hormone in pompano (Trachinotus ovatus).

Gonadotropin-releasing hormone (GnRH) is an important neuropeptide in the reproductive system. Although GnRH analogues have been used to artificially spawn pompano (Trachinotus sp.), the native forms of GnRH have not been described in this species. In this study three GnRH subtypes [sea bream GnRH (sbGnRH), chicken GnRH-Ⅱ (cGnRH-Ⅱ) and salmon GnRH (sGnRH)] were identified in pompano (Trachinotus ovatus). cgnrh-Ⅱ and sgnrh were mainly expressed in the brain of male and female fish, showing a tissue-specific expression pattern, while sbgnrh was expressed at different transcriptional levels in all tested tissues. In vivo injection experiment showed that sbGnRH significantly increased fsh and lh genes expression in a dose-dependent manner, but a high concentration of sbGnRH could desensitize the expression of lh. High concentrations of cGnRH-Ⅱ and sGnRH could induce the expression of fsh and lh. In addition, the results of in vitro incubation experiments showed that the high concentration of sbGnRH peptide could induce the expression of fsh and lh, while cGnRH-Ⅱ and sGnRH peptides could only induce the expression of fsh. 17ß-estradiol (E2) and 17α-methyltestosterone (MT) significantly inhibited sbgnrh mRNA expression in a dose-dependent manner, but did not affect the expression of cgnrh-Ⅱ and sgnrh mRNA. sbGnRH is the main GnRH subtype in pompano. E2 and MT can play a negative role in the regulation of sbgnrh. This study provides a theoretical basis for the reproductive endocrinology of pompano.

[Study on the association of IL-10-592 polymorphism with susceptibility to hepatitis B viral infection in Han, Yi and Yao ethnic groups in Guizhou province].

OBJECTIVE: To investigate the association of IL-10 gene promoter polymorphism with susceptibility to hepatitis B viral infection in Han, Yi and Yao ethnic groups from Guizhou province. METHODS: Five hundred volunteers from Guizhou province were selected to undertake PCR-RFLP for detection of IL-10 gene promoter -592 polymorphism. RESULTS: The genotypic distributions of IL-10-592 were 32.53%-51.43% (AA), 40.74%-54.82% (AC), 5.79%-18.52% (CC) whereas the allelic frequencies were 59.94%-72.38% for the A allele, and 27.62%-40.06% for the C allele in Han, Yi and Yao ethnic from Guizhou. The distributions of allele and genotype frequencies of IL-10-592 were statistically different between Yao ethnic in Libo and Yi ethnic in Qianxi, Yao ethnic in Libo and Han ethnic in Libo,Yi ethnic in Qianxi and Yi ethnic in Weining,Yi ethnic in Weining and Han ethnic in Libo (P < 0.05). IL-10-592 polymorphism was associated with HBV infection in Yi ethnic in Qianxi and the whole population. CONCLUSION: IL-10-592 gene polymorphisms influenced the susceptibility to HBV infection in Han, Yao, Yi sub-populations in Guizhou. Result of the study suggested that IL-10-592 gene polymorphisms might serve as a risk factor to HBV infection.

[Frequencies of GSTM1, GSTT1, and GSTP1 polymorphisms in ethnic minority from Guizhou].

OBJECTIVE: To investigate the frequencies of GSTM1, GSTT1 and GSTP1 polymorphisms in Dong, Yi and Yao ethnic groups from Guizhou. METHODS: In 321 volunteers who were population-based, GSTM1 and GSTT1 polymorphisms were analyzed by a multiplex-PCR procedure, whereas GSTP1 polymorphism was analyzed by PCR-RFLP method. RESULTS: Null genotype for GSTM1 and GSTT1 was 59.6%-71.2% and 39.4%-72.5%, respectively. The genotypic distribution of GSTP1 was 63.3%-75% for AA, 23.2%-35.8% for AG, 0-1.9% for GG, whereas the allelic frequencies were 81.2%-86.6% for the A allele, and 13.4%-18.8% for the G allele. CONCLUSION: There is a significant relationship between GSTT1 frequencies and ethnic populations.

[Protection of Tianshen Yizhi Recipe against low expression of nicotinic receptor and neurotoxicity induced by beta-amyloid peptide].

OBJECTIVE: To investigate the inhibition effects of Tianshen Yizhi Recipe (TSYZR), a compound traditional Chinese herbal medicine, on decreased expression of nicotinic acetylcholine receptor (nAChR) and the neurotoxicity as well as lipid peroxidation induced by beta-amyloid peptide (Abeta) in human SH-SY5Y neuroblastoma cells. METHODS: The SH-SY5Y cells were treated by a certain concentration of TSYZR, and then exposed to Abeta(25-35). Methyl thiazolyl tetrazolium reduction assay was carried out to understand the influences of the drugs on cellular viability. Expressions of nAChR subunits (alpha3 and alpha7) at protein and mRNA levels were detected by Western-blotting and reverse transcription polymerase chain reaction, respectively. Lipid peroxidation was measured by thiobarbituric acid to observe the capacity of antioxidant of the drugs. RESULTS: TSYZR at a safe concentration could increase alpha7 protein in the cells, inhibit decreased expressions of alpha3 and alpha7 nAChR subunit proteins, prevent lower expression of alpha7 mRNA in SH-SY5Y cells induced by Abeta, reduce the neurotoxicity and lipid peroxidation resulting from Abeta, but had no significant effect on the lower expression of alpha3 mRNA. CONCLUSIONS: TSYZR can up-regulate the expression of alpha7 nAChR subunit protein and prevent decreased expressions of nAChRs and neurotoxicity as well as lipid peroxidation induced by Abeta. This drug may play an important therapeutic role in treatment of Alzheimer disease.

[Polymorphism analysis of seven Y-STR loci in Shui population in Guizhou, China].

The objective of this study was to investigate the polymorphism of seven Y-specific STR loci in Shui ethnic population of Guizhou, China, and to obtain the polymorphism information in this minority. One trinucleotide STR locus and six tetranucleotide STR loci were simultaneously amplified with fluorescently labeled primers, and genotypes were determined with ABI PRISM 377 DNA Sequencer. Allele frequencies, genetic diversity and haplotype diversity were calculated. Among 94 unrelated males, 6, 4, 6, 2, 3, 5, 4 alleles were observed in loci DYS19, DYS389 I, DYS389 II, DYS390, DYS391, DYS392 and DYS393, respectively. Altogether, 27 haplotypes were identified for the seven Y-STR loci. The genetic diversity values for each locus ranged from 0.124 (DYS389 I) to 0.630 (DYS19). The haplotype diversity value was 0.868. High haplotype diversities were found in Shui population of Guizhou. The study suggests that these seven Y-STR loci are valuable Y-specific markers for establishing a Y-STR database, understanding ethical origin and migrations and for personal identification.

[Y-SNP analysis of Bouyei, Gelao, Mulao, Maonan, and Zhuang populations in Guizhou].

To understand the patrilineal genetic structure of Baiyue ethnic group in Guizhou province, we studied the frequencies of Y-chromosome haplotypes which consisted of 10 single nucleotide polymorphisms (SNPs) by using the PCR-RFLP method. Five haplotypes were found in Baiyue ethnic group in Guizhou, among which H8 was the most common, whereas that of Miao in Guizhou tended to be H8, H11 and H12 haplotypes. Compared with Miao in Guizhou, the Guizhou Baiyue (excluding Sui and Dong) was significantly different and could be regarded as an independent ethnic group. Differences were also found in the same ethnic group among different areas.

[Analysis of polymorphisms in Y-DNA haplotypes and mtDNA haplogroups in Yao ethnic group from Guizho].

The frequencies of Y-chromosome haplotypes consisting of 12 single nucleotide polymorphisms (SNP) and mitochondrial DNA (mtDNA) haplotypes consisting of 9 SNPs were investigated using PCR-RFLP in 97 Yaos from Guizhou to study the patrilineal and matrilineal genetic structure and the origin of Yao Ethnic Group from Guizhou. Results showed that all 97 samples were classified into 4 Y-DNA haplotypes (H7, H8, H9 and H11). The major haplotype H7 of Hmong-Mien Population was highly prevalent (92.4%) in Yaos from Guizhou. Eight mtDNA haplotypes were identified through mtDNA analysis and all the haplotypes could be classified into 5 haplogroups (B4, B5, D4, D5 and N*) that were defined previously. The frequency of a 9-bp deletion in the human mtDNA Co II/tRNALys intergenic region was 58.2% in the 97 samples. These data suggest that the patrilineal genetic structure of Yao ethnic group from Guizhou was simple and Yaos from Guizhou had the typical genetic character of the Hmong-Mien Population and some admixture with other populations. The matrilineal genetic structure of Yaos from Guizhou was relatively complex and the 9-bp deletion was a characteristic genetic marker of the matrilineal genetic structure of Yaos from Guizhou.

[G6PD Gene Mutations in Shui people in Sandu of Guizhou].

UNLABELLED: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzymopathy. To date, about 126 mutations in the G6PD gene have been detected, among which 17 mutations were found in Chinese. The most common mutations are: 1376 G-->T and 1388 G-->A, both in exon 12; 95 A-->G in exon 2, which amounted to more than 50% of mutations representing various regions and ethnic groups in China. A large-scale screening and genotypic analysis was held in Shui people in Sandu of Guizhou. To investigate the incidence and the molecular basis of G6PD deficiency of Guizhou Shui people, NBT qualitative and G6PD/6PGD quantitative methods were used to detect G6PD deficiency in 1,090 Shui people from the general people belonging to Sandu of Guizhou. By means of mis-matched primers amplified the G6PD gene, the products were 234 bp, 280 bp and 345 bp in length, then restriction enzyme analysis was used to detect the most common Chinese G6PD mutations, 1376 G-->T, 1388 G-->A and 95 A-->G. The results showed that out of the 1,090 samples, 98 G6PD deficiency samples were found. The incidence of G6PD deficiency was 8.99%. 24 cases of 1376 G-->T, 12 cases of 1388 G-->A, 9 cases of 95 A-->G were detected. A sample with 1376 G-->T and 95 A-->G mutation was found in a girl. It was reported for the first time. IN CONCLUSION: 1376 G-->T, 1388 G-->A, 95 A-->G mutations are the common G6PD mutations in Shui people in Sandu of Guizhou. The results indicates that different national minorities of Chinese may originated from a common ancestor.

[The sequence polymorphism of Y-chromosome DNA and mtDNA of Dong Ethnic of Congjiang, Guizhou].

To study the patrilineal and matrilineal genetic structure and the origin of Dong Ethnic of Congjiang Guizhou. Study the distribution of Y-chromosome haplotypes which consisted of 10 SNPs of Y-DNA and mtDNA haplogroups consisted of 11 SNPs by using PCR-RFLP method. The result is three haplotypes H6,H11,H14 were detected, the frequency of H11 is 92.5%. Six haplogroups were identified by mtDNA analysis, 75% of the people can be identified. The patrilineal genetic structure of Dong of Guizhou is simple, Principle component indicated that the structure is closer to Zhuang-Dong branch of Sino-Tibetan language family. The matrilineal genetic structure of Dong of Guizhou is complicated.

[The distribution of Y-chromosome haplotypes of Shui ethnic in Sandu,Guizhou].

Non-recombination region of Y-chromosome is a useful marker in tracing evolutionary history of paternal lineage. In the present study, total 92 individuals from Shui ethnic group in Sandu Shui Ethnic Group Autonomous County of Guizhou Province were inspected with 11 SNP sites including M7, M9, M15, M45, M89, M95, M119,M122, M130, M134 and YAP on Y-chromosome.All the subjects were required to be unrelated and without intermarriage with other ethnic groups within three generations. The haplotypes were analyzed by PCR-RFLP method. Four haplotypes H5,H8,H9 and H11 were detected with frequencies of 0.054, 0.044, 0.315 and 0.587, respectively.Principle component indicated that the paternal lineage of Shui ethnic group is much closer to Li ethnic group of Hainan Province and Bouyei ethnic group of Guizhou Province,which belong to the group of Zhuang-Dong branch of Sino-Tibetan language family. In addition genetic study of Shui coincides with its linguistic distribution.