Generation and analysis of expressed sequence tags from a cDNA library of the fruiting body of Ganoderma lucidum

<p>Abstract</p> <p>Background</p> <p>Little genomic or trancriptomic information on <it>Ganoderma lucidum </it>(<it>Lingzhi</it>) is known. This study aims to discover the transcripts involved in secondary metabolite biosynthesis and developmental regulation of <it>G. lucidum </it>using an expressed sequence tag (EST) library.</p> <p>Methods</p> <p>A cDNA library was constructed from the <it>G</it>. <it>lucidum </it>fruiting body. Its high-quality ESTs were assembled into unique sequences with contigs and singletons. The unique sequences were annotated according to sequence similarities to genes or proteins available in public databases. The detection of simple sequence repeats (SSRs) was preformed by online analysis.</p> <p>Results</p> <p>A total of 1,023 clones were randomly selected from the <it>G</it>. <it>lucidum </it>library and sequenced, yielding 879 high-quality ESTs. These ESTs showed similarities to a diverse range of genes. The sequences encoding squalene epoxidase (SE) and farnesyl-diphosphate synthase (FPS) were identified in this EST collection. Several candidate genes, such as <it>hydrophobin</it>, <it>MOB2</it>, <it>profilin </it>and <it>PHO84 </it>were detected for the first time in <it>G</it>. <it>lucidum</it>. Thirteen (13) potential SSR-motif microsatellite loci were also identified.</p> <p>Conclusion</p> <p>The present study demonstrates a successful application of EST analysis in the discovery of transcripts involved in the secondary metabolite biosynthesis and the developmental regulation of <it>G. lucidum</it>.</p

Gender Gaps in the Measurement of Public Opinion About Homosexuality in Cross-national Surveys: A Question-Wording Experiment

Measures of attitudes towards homosexuality in cross-national studies have received criticism for not being ‘gender-sensitive’. The current study used a split-ballot design allowing for separate analyses of the attitudes towards ‘gay men and lesbian women’, ‘gay men’, and ‘lesbian women’ in a pooled sample of 3,381 participants from Great Britain, Hungary, and Portugal. Analyses controlling for sociodemographics showed that differences in attitudes towards male and female targets were generally small and did not interact with the gender of the rater. In addition, results showed that men’s attitudes towards homosexuality were more strongly related to their gender ideology than women’s attitudes. Implications of these findings for cross-national studies measuring attitudes towards homosexuality are discussed

Iron isotope fractionation during skarn Cu-Fe mineralization

Fe isotopes have been applied to the petrogenesis of ore deposits. However, the behavior of iron isotopes in the mineralization of porphyry-skarn deposits is still poorly understood. In this study, we report the Fe isotopes of ore mineral separations (magnetite, pyrite, chalcopyrite and pyrrhotite ) from two different skarn deposits, i.e., the Tonglvshan Cu-Fe skarn deposit developed in an oxidized hydrothermal system and the Anqing Cu skarn deposit developed in a reduced hydro-thermal system. In both deposits, the Fe isotopes of calculated equilibrium fluids are lighter than those of the intrusions responsible for the skarn and porphyry mineralization, corroborating the “light-Fe fluid” hypothesis. Interestingly, chalcopyrite in the oxidized-Tonglvshan skarn deposit has lighter Fe than chalcopyrite in the reduced-Anqing skarn deposit, which is best understood as the result of the prior precipitation of magnetite (heavy Fe) from the ore fluid in the oxidized-Tonglvshan systems and the prior precipitation of pyrrhotite (light Fe) from the ore fluid in the reduced-Anqing system. The δ 56Fe for pyrite shows an inverse correlation with δ 56Fe of magnetite in the Tonglvshan. In both deposits, the Fe isotope fractionation between chalcopyrite and pyrite is offset from equilibrium line at 350℃ and lies between the FeS-chalcopyrite equilibrium line and pyrite-chalcopyrite equilibrium line at 350℃. These observations are consistent with the FeS pathway towards pyrite formation. That is, Fe isotopes fractionation during pyrite formation depends on a path, from the initial FeS-fluid equilibrium towards the pyrite-fluid equilibrium due to the increasing extent of Fe isotopic exchange with fluids. This finding, together with the data from other deposits, allows us to propose that the pathway effect of pyrite formation in the Porphyry-skarn deposit mineralization is the dominant mechanism that controls Fe isotope characteristics

De novo sequencing and analysis of the American ginseng root transcriptome using a GS FLX Titanium platform to discover putative genes involved in ginsenoside biosynthesis

<p>Abstract</p> <p>Background</p> <p>American ginseng (<it>Panax quinquefolius </it>L.) is one of the most widely used herbal remedies in the world. Its major bioactive constituents are the triterpene saponins known as ginsenosides. However, little is known about ginsenoside biosynthesis in American ginseng, especially the late steps of the pathway.</p> <p>Results</p> <p>In this study, a one-quarter 454 sequencing run produced 209,747 high-quality reads with an average sequence length of 427 bases. <it>De novo </it>assembly generated 31,088 unique sequences containing 16,592 contigs and 14,496 singletons. About 93.1% of the high-quality reads were assembled into contigs with an average 8-fold coverage. A total of 21,684 (69.8%) unique sequences were annotated by a BLAST similarity search against four public sequence databases, and 4,097 of the unique sequences were assigned to specific metabolic pathways by the Kyoto Encyclopedia of Genes and Genomes. Based on the bioinformatic analysis described above, we found all of the known enzymes involved in ginsenoside backbone synthesis, starting from acetyl-CoA via the isoprenoid pathway. Additionally, a total of 150 cytochrome P450 (CYP450) and 235 glycosyltransferase unique sequences were found in the 454 cDNA library, some of which encode enzymes responsible for the conversion of the ginsenoside backbone into the various ginsenosides. Finally, one CYP450 and four UDP-glycosyltransferases were selected as the candidates most likely to be involved in ginsenoside biosynthesis through a methyl jasmonate (MeJA) inducibility experiment and tissue-specific expression pattern analysis based on a real-time PCR assay.</p> <p>Conclusions</p> <p>We demonstrated, with the assistance of the MeJA inducibility experiment and tissue-specific expression pattern analysis, that transcriptome analysis based on 454 pyrosequencing is a powerful tool for determining the genes encoding enzymes responsible for the biosynthesis of secondary metabolites in non-model plants. Additionally, the expressed sequence tags (ESTs) and unique sequences from this study provide an important resource for the scientific community that is interested in the molecular genetics and functional genomics of American ginseng.</p

Association between daily screen time and risk of stroke among middle-aged and elderly people: research based on China health and nutrition survey

BackgroundWe aimed to explore the independent associations between screen time and the risk of stroke among Chinese adults based on the China Health and Nutrition Survey (CHNS).MethodsData on Chinese adults aged older than 40 years from the CHNS in during 2004–2009 were selected. A total of 4,587 individuals were included in 2009, including screen time and the risk of stroke. Simultaneously, we traced the previous screen time to 2004 for those with outcome measures in 2009 (n = 2,100). Basic information, lifestyle, and screen behavior were obtained through face-to-face interviews and self-completed questionnaires. Anthropometric data collected included blood pressure, body weight, height, hip circumference, and waist circumference. Fasting blood was obtained for measurements of lipid and glucose levels. Cross-sectional analysis and cohort analysis were both performed using multivariate logistic regression.ResultsOf all participants, 3,004 (65.49%) participants spent more than 2 h per day on screen time. Taking the men who spent less than 2 h on screen per day as reference, the crude odds ratio (OR) of the high risk of stroke was 1.53 [95% confidence interval (CI), 1.20–1.95] for the men who spent 2–3 h per day on screen and 2.37 (95% CI, 1.78–3.16) for the men who spent more than 3 h per day on screen. This difference remained significant after adjusting for confounding factors. No association was observed among women. However, in the cohort analysis with screen time in 2006 as the independent variable, the association between screen time and stroke risk was found both in men [OR, 1.83 (95% CI, 1.19–2.82)] and women [OR, 1.48 (95% CI, 1.10–1.99)]).ConclusionWe found that the high screen time was associated with an increased stroke risk, which was pronounced in men, warranting a universal need to limit screen time in order to improve health

Clinical report about the effectiveness of andriol, tamoxifen, vitamin A and lecithin on low acrosome

BackgroundAcrosin is the most important proteolytic enzyme in fertilization, acrosome dysfunction affects fertilization. Recently, our group found that the treatment of andriol, tamoxifen, vitamin A and lecithin helped to improve acrosome function.AimsTo assess the effectiveness of andriol, tamoxifen, vitamin A and lecithin on low acrosome.MethodsSeventy-eight infertility males in my hospital were enrolled. Kenedy test was used to test acrosome function. Andriol, tamoxifen, vitamin A and lecithin were used for one to four months.ResultsSperm concentration (t=5.05, P=0.000) raised while seminal volume (t=1.93, P=0.057), normal morphology (t=0.24, P=0.811) and progressive sperm (t=2.14, P=0.036) did not show statistical difference after treatment. Acrosin was (37.62±11.07)μIU/106 sperm before treatment and raised to (52.68±15.68)μIU/106 sperm after treatment and showed statistical difference (t=8.18, P=0.00). In all the 78 males, the acrosin of seven men (11 per cent) bellowed after treatment while the acrosin of 71 males (91 per cent) raised and ten couples (13 per cent) got pregnant.ConclusionThe combined treatment of andriol, tamoxifen, vitamin A and lecithin were effective on low acrosin

Effectiveness of two kind of medicine on non-obstructive azoospermia

ObjectiveTo assess the effectiveness of medicine on non-obstructive azoospermia (NOA). Cases32 male patients were enrolled from March 2012 to March 2016, with age range of 23–41 years old and infertility time of 1–3 years.Methods According to “WHO Human Semen Detection and Processing Experiment Manual";;;;;; fifth edition, semen routine was based on manual detection and serum hormones based on chemiluminescence detection, semen volume, sperm density, forward moving sperm ratio. If sperm was detected in semen for more than two consecutive times, the treatment was considered effective and was included into the statistical analysis. The detection parameters took the first test data and the last test data. Treatment Andriol and tamoxifen were used for idiopathic NOA while aescuven forte and aspirin used for NOA with varicocele (including two varicocelectomy). The time was four months.ResultsOf the 32 patients, 13 underwent testicular biopsy, pathological results showed six patients (46 per cent) with hypospermatogenesis, five patients (39 per cent) with maturation arrest, and two patients (15 per cent) with sertoli cell syndrome. Nine patients (28 per cent) had varicocele; two patients (6 per cent) underwent surgical treatment. After treatment, sperm was detected in 20 patients (63 per cent), seven of them (22 per cent) had detectable sperm in semen after one month of treatment; five (19 per cent) had sperm after two months of treatment; eight (25 per cent) had sperm after three months of treatment. Among the 23 patients with idiopathic azoospermia, 13 patients had detectable sperm in semen after treatment (56 per cent). The level of hormone gradually increased in 15 patients, rapidly increased in four patients, but four patients had no obvious change of hormone level. Among the nine patients with varicocele, three persons (including one patient undergoing surgery) had normal sperm density after 3–4 months of treatment, continuing the treatment for another 2–3 months, wife of the patients gained pregnancies. After treatment, seven patients had gradually decreasing serum level of FSH, LH and T while two patients had increased serum level of hormones. Excluding the three pregnancy patients, it was showed that the sperm density increased (t=-3.342, p=0.004) after treatment, so did the forward moving sperm ratio (t=-4.037, p=0.001), but semen volume had no significant changes (t=-1.698, p=0.109). ConclusionThe treatment of varococele is important for the NOA with varicocele while andriol and tamoxifen is other method for the treatment of idiopathic NOA