Analysis of microbial diversity in the root of Astragalus mongholicus.

The dry root of Astragalus mongholicus has therapeutic effects such as tonifing the middle - jiao, replenishing qi, solidifing the surface, promoting diuresis, dispelling sepsis outwards and nourishing muscle. There are some slices having black spots after slicing the root of astragalus. The diversity of endophytic fungi between slices with black spots and normal slices was analysed in this paper. The endophytic fungal sequences obtained by high-throughput sequencing were 298,044 and 297,396, and the 116 OTU subsets obtained after clustering belonged to 3 phyla, 9 classes, 22 orders, 38 families and 46 genera. The dominant classes were Eurotiomycetes and Leotiomycetes. The dominant order is Eurotiales and Helotiales. The dominant families are Helotiales_fam_Incertae_sedis and Aspergillaceae. The dominant genera are Cadophora and Aspergillus. There are some peculiar fungal flora in both normal slices and spotted slices. The study on endophytic fungi diversity of astragalus slices will provide some help for drug development of this plant.

Analysis of microbial diversity in the root of Astragalus mongholicus / Análise da diversidade microbiana em raízes de astrágalo

The dry root of Astragalus mongholicus has therapeutic effects such as tonifing the middle - jiao, replenishing qi, solidifing the surface, promoting diuresis, dispelling sepsis outwards and nourishing muscle. There are some slices having black spots after slicing the root of astragalus. The diversity of endophytic fungi between slices with black spots and normal slices was analysed in this paper. The endophytic fungal sequences obtained by high-throughput sequencing were 298,044 and 297,396, and the 116 OTU subsets obtained after clustering belonged to 3 phyla, 9 classes, 22 orders, 38 families and 46 genera. The dominant classes were Eurotiomycetes and Leotiomycetes. The dominant order is Eurotiales and Helotiales. The dominant families are Helotiales_fam_Incertae_sedis and Aspergillaceae. The dominant genera are Cadophora and Aspergillus. There are some peculiar fungal flora in both normal slices and spotted slices. The study on endophytic fungi diversity of astragalus slices will provide some help for drug development of this plant.

A raiz seca de astrágalo tem a função de tonificar o zhongjiao, qi benéfico, superfície sólida, diurético, remoção externa de veneno e fortalecimento muscular. Depois de cortar o astrágalo em fatias finas, algumas fatias têm pontos pretos. Neste trabalho foi analisada a diversidade de fungos endofíticos em fatias de ponto negro versus fatias normais. O sequenciamento de alto rendimento resultou em 314.998 e 315.051 sequências de fungos endofíticos, respectivamente, que após agrupamento resultaram em 116 subgrupos de OTU, pertencentes a três filos, nove classes, 22 ordens, 38 famílias e 46 gêneros. As classes dominantes são Eurotiomycetes e Leotiomycetes. A ordem principal é euclides e hilllows. As famílias predominantes foram aspergilaceae e aspergilaceae. Os gêneros predominantes foram leucostilla e aspergillus. Tanto as seções normais como as secções manchadas apresentam uma flora fúngica distinta. O estudo da diversidade de fungos endofíticos em fatias de astrágalo pode contribuir para o desenvolvimento de fármacos para esta planta.

Identification of genes related to floral organ development in pak choi by expression profiling.

Pak choi is a highly nutritious vegetable that is widely grown in China, Southeast Asia, and other parts of the world. Because it reproduces by seed, it is very important to understand the mechanism of floral organ development. Therefore, using the Chinese cabbage genome as a reference, this study analyzed the expression profiles of shoot apex genes at flower bud differentiation stages 1 and 5, in order to identify genes related to floral organ development. The results showed that the proportion of mapped genes was high, with 84.25 and 83.80% of clean reads from the two sample saligned to the reference genome, respectively. A total of 525 differentially expressed genes (DEGs) were identified, 224 of which were upregulated and 301 were downregulated. The expression levels of genes homologous to Chinese cabbage flowering genes were also analyzed at stages 1 and 5; the expression levels of Bra012997 (ap1), Bra000393 (SOC1), and Bra004928 (SOC1) were significantly upregulated at stage 5, suggesting that these three genes positively regulate floral development in pak choi. DEGs involved in floral organ development were analyzed with homologous genes from Arabidopsis thaliana; the homologous genes Bra029281 (AGL42), Bra026577 (ARPN), Bra022954 (SPL3), Bra029293 (ARF2), Bra007978 (AtRLP12), Bra033221 (SPL8), Bra008037 (LOX4), Bra001598 (IAA19), Bra003892 (PATL1), Bra038778 (AT4G21323), Bra025315 (KLCR2), and Bra013906 (DTX35) are directly related to floral organ development in Arabidopsis, suggesting that these genes have corresponding functions during flower organ development in pak choi, and could be candidates for further genetic research. These results provide a foundation for research on the molecular mechanism of flower organ development in pak choi and other Brassica rapa vegetables.

Microbial diversity in raw milk and traditional fermented dairy products (Hurood cheese and Jueke) from Inner Mongolia, China.

Hurood cheese (HC) and Jueke (Jk) are 2 traditional fermented dairy products produced from raw milk (RM) in the Inner Mongolia region of China. They have a long history of production and consumption. The microbial compositions of RM, HC, and Jk vary greatly, and are influenced by their geographical origins and unique processing methods. In this study, 2 batches of RM, HC, and Jk samples were collected (April and August 2015) from the Zhenglan Banner, a region located in the southern part of Inner Mongolian belonging to the Xilingol league prefecture. The bacterial and fungal diversities of the samples were determined by 16S rRNA and 18S rRNA gene sequence analysis, respectively. A total of 112 bacterial and 30 fungal sequences were identified, with Firmicutes and Ascomycota being the predominant phyla for bacteria and fungi, respectively. Lactococcus and Lactobacillus were identified as the main bacterial genera, whereas Kluyveromyces was the predominant fungus identified in the 3 dairy products. Different bacterial and fungal compositions were observed in RM, HC, and Jk samples collected at different times. These results suggested that time of production may be an important factor influencing the microbial diversity present in RM, HC, and Jk.

MYBA2 gene involved in anthocyanin and flavonol biosynthesis pathways in grapevine.

MYBA2 transcription factor (Myb-related gene) affects the coloring in grapevine berry and plays an important role in the biosynthesis of anthocyanin. The MYBA2 gene was cloned from Vitis vinifera L. cv. Cabernet Sauvignon and polyclonal antibodies for VvmybA2 were prepared. The VvmybA2 gene expression patterns were observed in seven tissues (the leaf, stem, flower, bud, root, berry, and tendril) and during the berry development stage at transcriptional and translational levels, respectively. The results indicated that the expression of VvmybA2 was approximately 11-fold higher in the berry than that in the other six tissues, and increased rapidly from 60 days after full bloom reaching a maximum on day 80. Furthermore, both the anthocyanin content and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT) gene expression levels increased rapidly 60 days after full bloom. Moreover, correlation analysis indicated that the transcriptional and translational expression levels of the VvmybA2 gene were significantly positively correlated with not only UFGT and DFR genes but also with the anthocyanin content during berry development. These results suggested that VvmybA2 could not only regulate the transcription of both UFGT and DFR but also is involved in the expression of the UFGT gene associated with color determination in grape berries.

Association between TNF-α rs1799724 and rs1800629 polymorphisms and the risk of Crohn's disease.

We investigated the associations between 2 major tumor necrosis factor-α (TNF-α) polymorphisms, rs1799724 C>T and rs1800629 G>A, and the susceptibility to Crohn's disease (CD) using a meta-analysis framework. The PubMed, EBSCO, Ovid, Wiley, Web of Science, WANFANG, and VIP databases (last updated search in October 2014) were comprehensively searched for relevant published studies. The studies retrieved from database searches were filtered based on inclusion and exclusion criteria, and the resultant data extracted from the selected studies were analyzed using the Comprehensive Meta-analysis 2.0 software. Eleven case-control studies, containing 2000 CD patients and 3499 healthy controls, were identified as relevant to this meta-analysis. Data extracted from these 11 studies were analyzed to understand the role of the 2 TNF-α polymorphisms in CD. We found that the TNF-α rs1799724 C>T polymorphism increased the susceptibility to CD (allele model: OR = 1.293, 95%CI = 1.090-1.534, P = 0.003; dominant model: OR = 1.258, 95%CI = 1.031-1.534, P = 0.024). In contrast, we found no significant association between the TNF-α rs1800629 G>A polymorphism and CD susceptibility (allele model: OR = 1.005, 95%CI = 0.864-1.170, P = 0.945; dominant model: OR = 0.962, 95%CI = 0.809-1.145, P = 0.667). This meta-analysis showed that the TNF-α rs1799724 C>T polymorphism is associated with CD susceptibility, while the TNF-α rs1800629 G>A polymorphism appeared to have no correlation with the susceptibility to CD.

Effect of NB-UVB on levels of MCP-1 and CCR6 mRNA in patients with psoriasis vulgaris.

The aim of this study is to explore the effect of narrow band ultraviolet B (NB-UVB) on the chemokine receptor CCR6 mRNA levels in patients with psoriasis. Psoriasis area and severity index (PASI) values were recorded before and after the treatment with NB-UVB phototherapy of 30 psoriasis vulgaris patients. The reverse transcription-polymerase chain reaction method was used to detect the expression level of CCR6 mRNA in peripheral blood mononuclear cells, and compared with 30 healthy subjects. The PASI value of the 30 psoriasis vulgaris patients decreased significantly after 15 iterations of phototherapy treatment (P < 0.01). The expression level of CCR6 mRNA in psoriasis patients was significantly higher than in the healthy controls (P < 0.01), while the expression level of CCR6 mRNA decreased significantly after phototherapy (P < 0.01). Reduction of CCR6 level may be one of the mechanisms through which NB-UVB can treat psoriasis.

Accumulation of flavanols and expression of leucoanthocyanidin reductase induced by postharvest UV-C irradiation in grape berry.

To examine the effect of postharvest ultraviolet C (UV-C) irradiation on flavanol polyphenol accumulation in the grape berry, we investigated total flavanol polyphenol content, the enzyme activity of leucoanthocyanidin reductase (LAR), and transcription of Vv lar1 and Vv lar2 using spectrophotometry, real-time polymerase chain reaction, and western blot analysis in 5-year-old Vitis vinifera L. cv. Cabernet Sauvignon plants. Our results indicated that the accumulation of flavanol polyphenol reached its highest value when exposed to UV-C irradiation for 30 min. Additionally, UV-C irradiation induced the transcription of Vv lar1 and Vv lar2 and the synthesis of LAR1 and LAR2 proteins, resulting in increased accumulation of flavanol polyphenol in the grape berry. Moreover, these effects were associated with the length of time of UV-C irradiation.

Analysis of regulatory T cells frequency in peripheral blood and tumor tissues in papillary thyroid carcinoma with and without Hashimoto's thyroiditis.

PURPOSE: Regulatory T cells (Treg) suppress the immune reaction. The aim of the present study was to investigate the clinicopathologic significance and roles of Treg in papillary thyroid carcinoma (PTC) patients with and without Hashimoto's thyroiditis. METHODS: Flow cytometry was used to detect the percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells in peripheral blood. FoxP3+ Treg were detected by immunohistochemistry in the tumor tissues. RESULTS: The percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells was significantly higher in PTC patients than that in multinodular goiter (MNG) patients. There were large numbers of tumor-infiltrating FoxP3+ Treg in primary PTC and metastatic lymph nodes tissues; however, there was no FoxP3 expression in the MNG tissues. Higher percentage of Treg both in peripheral blood and tumor tissues was associated with extrathyroidal extension and lymph nodes metastasis. The percentage of CD4+CD25+CD127low/- Treg among CD4+ T cells in peripheral blood of PTC patients with Hashimoto's thyroiditis (HT) was significantly lower, whereas the infiltration of FoxP3+ Treg in tissues of PTC with HT tended to be increased. CONCLUSIONS: We concluded that the percentage of Treg increased in peripheral blood as well as in the tumor tissues of PTC patients compared with that of MNG patients. The high percentage of Treg was associated with aggressiveness. There may be a compensatory expansion of Treg at the sites of inflammation in tissues of PTC with HT contributing to the immune response suppression.

Dissecting the mechanism of Solanum lycopersicum and Solanum chilense flower colour formation.

Flowers are the defining feature of angiosperms, and function as indispensable organs for sexual reproduction. Flower colour typically plays an important role in attracting pollinators, and can show considerable variation, even between closely related species. For example, domesticated tomato (S. lycopersicum) has orange/yellow flowers, while the wild relative S. chilense (accession LA2405) has bright yellow flowers. In this study, the mechanism of flower colour formation in these two species was compared by evaluating the accumulation of carotenoids, assessing the expression genes related to carotenoid biosynthetic pathways and observing chromoplast ultrastructure. In S. chilense petals, genes associated with the lutein branch of the carotenoid biosynthetic pathway, phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), lycopene ß-cyclase (LCY-B), ß-ring hydroxylase (CRTR-B) and ε-ring hydroxylase (CRTR-E), were highly expressed, and this was correlated with high levels of lutein accumulation. In contrast, PDS, ZDS and CYC-B from the neoxanthin biosynthetic branch were highly expressed in S. lycopersicum anthers, leading to increased ß-carotene accumulation and hence an orange/yellow colour. Changes in the size, amount and electron density of plastoglobules in chromoplasts provided further evidence of carotenoid accumulation and flower colour formation. Taken together, these results reveal the biochemical basis of differences in carotenoid pigment accumulation and colour between petals and anthers in tomato.

Development of eight novel microsatellite markers for Huoyan geese.

In this study, we isolated microsatellite DNA from the Huoyan goose genome with magnetic beads. As a result, 150 positive clones were identified, and 148 microsatellites were found. Among the 148 microsatellites, 69.6% were perfect, 17.6% were imperfect, and the rest were compound type (12.8%). Twenty microsatellite primers were used to screen 90 individuals from 3 Huoyan goose populations. Eight loci were polymorphic with a low number of alleles (2 to 4). The observed and expected heterozygosities ranged from 0.3556 to 1 and from 0.2923 to 0.6868, respectively. All the 8 polymorphic loci were in Hardy-Weinberg equilibrium. These molecular markers will be useful for future studies on population genetic structure and conservation genetics in Huoyan geese.

Comparative study between Ephedra sinica Stapf and Fructus Schisandrae Chinensis on ET-1 and 6-keto-prostaglandin F1α in rats with idiopathic pulmonary fibrosis.

The aim of this study was to evaluate differences in the effects of Ephedra sinica Stapf and Fructus Schisandrae Chinensis on angiogenesis in the treatment of bleomycin-induced rat idiopathic pulmonary fibrosis. The rat models were created using bleomycin. The animals were divided into six groups: model, control, Ephedra alone, Schisandrae alone, combination of Ephedra and Schisandrae, and hydrocortisone alone. The treatments were administered for 28 days. After 7 and 28 days, the rats were sacrificed for pathological morphology examination, microvascular density determination, and angiogenesis-related cytokine examination. The Ephedra and hydrocortisone groups demonstrated significantly reduced alveolitis and pulmonary fibrosis grades compared with the model group (P < 0.05). The number of blood vessels in the Ephedra group was higher than that in the Schisandrae and combination therapy groups. At 7 days, the expression level of endothelin (ET)-1 in the model group was significantly higher than that in the normal group (P < 0.01). The level of 6-keto-prostaglandin F1α (6-keto-PGF1α) in the treatment group increased, and there were significant differences between the Ephedra group and the combination therapy and normal groups (P < 0.05). Ephedra inhibited the increase in the lung coefficient. The combination therapy prevented pulmonary artery injury and angiogenesis of the arteries by reducing the level of ET-1 and promoting the level of 6-keto-PGF1α in the blood. Ephedra and Schisandrae prevented alveolitis and the development of pulmonary fibrosis.

Genetic differentiation between natural and hatchery populations of Manila clam (Ruditapes philippinarum) based on microsatellite markers.

Manila clam (Ruditapes philippinarum) is one of the major aquaculture species around the world and supports an important segment of the aquaculture industry in China. In this study, we used ten microsatellite markers to detect genetic diversity within six R. philippinarum populations and genetic differentiation between them. A total of 109 alleles were detected across all loci. Compared to wild populations (N(A) = 8.4-9.1 alleles/locus, H(E) = 0.75-0.77, H(O) = 0.67-0.73), hatchery stocks showed less genetic variation as revealed in lower number of alleles and lower heterozygosity (N(A) = 7.4-7.5 alleles/locus, H(E) = 0.72-0.75, H(O) = 0.68-0.70), indicating that a bottleneck effect has occurred in hatchery history. Significant genetic differentiation was observed between cultured stocks (P < 0.05), and between cultured and wild populations (P < 0.05). Phylogenetic analysis showed a clear separation of the northern three populations and the southern three populations, suggesting that geographically separated populations of R. philippinarum could be genetically differentiated with limited genetic information exchanged between them. The information obtained in this study indicates that the northern and southern populations of R. philippinarum should be managed separately in hatchery practices for the preservation of genetic diversity in wild populations.

Expression of a GDP-L-galactose phosphorylase-like gene in a Chinese wild Vitis species induces responses to Erysiphe necator and defense signaling molecules.

Using rapid amplification of cDNA ends, a full-length cDNA sequence of a GDP-L-galactose phosphorylase-like gene was isolated from leaves infected by Erysiphe necator in the Chinese wild (Vitis pseudoreticulata) clone, 'Baihe-35-1', an E. necator-resistant genotype. The full-length cDNA, designated as VpVTC, comprised 1943 bp and putatively encodes a 453-amino acid polypeptide containing an HIT motif. The deduced amino acid sequence showed high similarity with that of VTC genes from other plants. The expression of VpVTC, determined by reverse transcriptase-polymerase chain reaction, was induced by E. necator and defense signaling molecules, including salicylic acid, methyl jasmonate, and ethephon, in 'Baihe-35-1', the V. quinquangularis genotype 'Shang-24', and the E. necator-susceptible V. pseudoreticulata genotype, 'Hunan-1'. Transcript levels of VpVTC correlated well with the degree of disease resistance in the 3 genotypes. Maximum induction of VpVTC by E. necator (>7-fold at 96 h post-inoculation) occurred in 'Baihe-35-1', which also showed the fastest response to signaling molecules. Upregulating the expression of VpVTC in 'Baihe-35-1' resulted in a gradual increase in the ascorbic acid concentration of leaves inoculated with E. necator. Furthermore, VpVTC was expressed in leaves, stems, inflorescence, tendrils, and fruit at all developmental stages, with the highest level occurring in fruit 35 days after flowering.

Genetic analysis of differentiation of T-helper lymphocytes.

In the human immune system, T-helper cells are able to differentiate into two lymphocyte subsets: Th1 and Th2. The intracellular signaling pathways of differentiation form a dynamic regulation network by secreting distinctive types of cytokines, while differentiation is regulated by two major gene loci: T-bet and GATA-3. We developed a system dynamics model to simulate the differentiation and re-differentiation process of T-helper cells, based on gene expression levels of T-bet and GATA-3 during differentiation of these cells. We arrived at three ultimate states of the model and came to the conclusion that cell differentiation potential exists as long as the system dynamics is at an unstable equilibrium point; the T-helper cells will no longer have the potential of differentiation when the model reaches a stable equilibrium point. In addition, the time lag caused by expression of transcription factors can lead to oscillations in the secretion of cytokines during differentiation.