miR-129b suppresses cell proliferation in the human lung cancer cell lines A549 and H1299.

Lung cancer is one of the most prevalent malignant tumors, and is one of the primary causes of cancer-associated deaths. In 2002, an estimated 1.18 million lung cancer-associated deaths were recorded, accounting for 18% of cancer-related deaths and 2% of total mortality. Despite the great progress that has been made in lung cancer therapies, the mechanisms underlying lung cancer formation and development remain largely unknown. Meanwhile, the microRNA miR-129 has been shown to be involved in the formation of many types of cancer. Therefore, this study aims to investigate whether miR129b could suppress proliferation of lung cancer cell lines. NSCLC tissue samples were collected from the Department of Respiratory Medicine between April 2013 and December 2015. Ten normal health individuals were recruited as controls. Lung cancer cell lines A549 and H1299 were used to examine the suppressive effects of miR129b. Quantitative real-time PCR was used to detect miR129b expression. The MTT assay was used to analyze cell proliferation. Results indicated that miR-129b is down-regulated in lung cancer cell lines and NSCLC tissues. Furthermore, overexpression of miR-129b inhibited proliferation of lung cancer cells. In conclusion, miR-129b suppresses lung cancer cell proliferation, and can be a potential therapeutic target for treatment of lung cancers.

Microsatellite analysis of genetic diversity in quail populations from China

Polymorphism of three quail communities was analyzed by using 12 microsatellite markers in this paper, aiming to provide scientific references for the evaluation, protection and utilization of quail genetic resources in China. Results demonstrated that the number of observed alleles by 12 microsatellite markers ranges between 4~7. The average polymorphism information contents (PIC) of the Chinese yellow quail, the Chinese black quail and the Korean quail, as detected by 12 microsatellite markers, are 0.6853, 0.6401 and 0.6565,respectively, and average heterozygosity values are 0.7333, 0.6957 and 0.7111, respectively. This indicates that the Chinese yellow quail has the richest genetic polymorphism. According to cluster analysis, the Chinese black quail and the Korean quail have the smallest genetic distance (0.0628), which reflects that they have the closest genetic relationship. The genetic distance between the Chinese yellow quail and the Korean quail is 0.0951. Therefore, the Chinese black quail and the Korean quail are clustered together firstly, and then the Chinese yellow quail.(AU)

Microsatellite analysis of genetic diversity in quail populations from China

Polymorphism of three quail communities was analyzed by using 12 microsatellite markers in this paper, aiming to provide scientific references for the evaluation, protection and utilization of quail genetic resources in China. Results demonstrated that the number of observed alleles by 12 microsatellite markers ranges between 4~7. The average polymorphism information contents (PIC) of the Chinese yellow quail, the Chinese black quail and the Korean quail, as detected by 12 microsatellite markers, are 0.6853, 0.6401 and 0.6565,respectively, and average heterozygosity values are 0.7333, 0.6957 and 0.7111, respectively. This indicates that the Chinese yellow quail has the richest genetic polymorphism. According to cluster analysis, the Chinese black quail and the Korean quail have the smallest genetic distance (0.0628), which reflects that they have the closest genetic relationship. The genetic distance between the Chinese yellow quail and the Korean quail is 0.0951. Therefore, the Chinese black quail and the Korean quail are clustered together firstly, and then the Chinese yellow quail.

Development and application of Est-Ssr markers in quails

Aiming at accelerating the application of molecular markers in the genetic improvement of quails, six EST-SSR markers were successfully developed using a bioinformatics method. Polymorphisms of three quail populations (Chinese yellow quail, China black quail and Korean quail) were detected. The results showed that there were 2-6 alleles in six EST-SSR markers. Mean polymorphism information contents of Chinese yellow quails, Chinese black quails and Korean quails were determined as 0.5451, 0.4962 and 0.4937, respectively. Average heterozygosity valuesof 0.6134, 0.5759 and 0.5613 were calculated. Among the six EST-SSR markers, three were highly polymorphic, and the other three were moderately polymorphic. The newly-developed six EST-SSR markers may be used to determine the genetic diversity of quails. The six EST-SSR markers identified were related to carbohydrate metabolism and melanin synthesis, but their specific mechanisms need to be further analyzed.(AU)

Development and application of Est-Ssr markers in quails

Aiming at accelerating the application of molecular markers in the genetic improvement of quails, six EST-SSR markers were successfully developed using a bioinformatics method. Polymorphisms of three quail populations (Chinese yellow quail, China black quail and Korean quail) were detected. The results showed that there were 2-6 alleles in six EST-SSR markers. Mean polymorphism information contents of Chinese yellow quails, Chinese black quails and Korean quails were determined as 0.5451, 0.4962 and 0.4937, respectively. Average heterozygosity valuesof 0.6134, 0.5759 and 0.5613 were calculated. Among the six EST-SSR markers, three were highly polymorphic, and the other three were moderately polymorphic. The newly-developed six EST-SSR markers may be used to determine the genetic diversity of quails. The six EST-SSR markers identified were related to carbohydrate metabolism and melanin synthesis, but their specific mechanisms need to be further analyzed.

Expression of MMP-1, -2, and -8 in longissimus dorsi muscle and their relationship with meat quality traits in cattle.

The extracellular matrix (ECM) is the major macromolecule in skeletal muscle, which affects meat quality greatly. The remodeling of the ECM is mainly regulated by matrix metalloproteinases (MMPs). The expression patterns of MMP-1, -2, and -8 in longissimus dorsi muscle were explored using quantitative real-time polymerase chain reaction. The results show that the expression of MMP-1, -2, and -8 decreased significantly from 135 days of pregnancy to postnatal 30 months. While the expression of MMP-1, -2, and -8 showed no significant relationships with intramuscular fat contents, MMP-1 and -2 showed significant negative correlations with the shearing force of the longissimus dorsi muscle in cattle. The expression of MMP-1 also showed a significant negative correlation with cooking loss and a positive correlation with water holding capacity. The expression levels of MMP-1 and -2 were usually higher in fat than in skeletal muscle tissue. The expression of MMP-8 was significantly higher in the mammary fat pad and the longissimus dorsi muscle than in all other tissues. This study indicates that the remodeling of the ECM has important effects both on the development of postnatal skeletal muscle and on meat quality.

Expression of ADAMTS4 and ADAMTS5 in longissimus dorsi muscle related to meat tenderness in Nanyang cattle.

The ADAMTS4 and ADAMTS5 are secreted proteases, which can cleave aggrecan, brevican and versican to regulate rebuilding of the extracellular matrix. We analyzed the ADAMTS4 and ADAMTS5 gene expression patterns in longissimus dorsi muscle at intervals from 135 days fetal age to 30 months old by qRT-PCR in Nanyang cattle. Expression of ADAMTS4 was significantly higher in 135 and 185-day-old fetuses than at other stages, while expression of ADAMTS5 decreased during development. The promoter regions of ADAMTS4 and ADAMTS5 were cloned and the transcription factor binding sites were analyzed with bioinformatic methods. Twelve and six potential transcription factor binding sites were found in the promoter regions of ADAMTS4 and ADAMTS5 genes, respectively. Three transcription factors (MZF1, C/EBPb, and NF-kap) were selected to analyze the expression pattern during the development of the longissimus dorsi muscle. MZF1 was significantly co-expressed with ADAMTS4, while C/EBPb expression was significantly negatively associated with that of ADAMTS4. We concluded that the expression of ADAMTS4 is positively regulated by MZF1 and negatively regulated by C/EBPb. We examined the relationships of ADAMTS4 and ADAMTS5 expression with tenderness of longissimus dorsi muscle; ADAMTS4 was significantly and negatively correlated with meat tenderness. We conclude that ADAMTS4 participates in the regulation of muscle development in cattle.

Separation and identification of Musa acuminate Colla (banana) leaf proteins by two-dimensional gel electrophoresis and mass spectrometry.

To establish a proteomic reference map of Musa acuminate Colla (banana) leaf, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 44 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. Three spots that were not identified by MALDI-TOF MS analysis were identified by searching against the NCBInr, SwissProt, and expressed sequence tag (EST) databases. We identified 41 unique proteins. The majority of the identified leaf proteins were found to be involved in energy metabolism. The results indicate that 2D-PAGE is a sensitive and powerful technique for the separation and identification of Musa leaf proteins. A summary of the identified proteins and their putative functions is discussed.

Isolation and characterization of nucleotide-binding site and C-terminal leucine-rich repeat-resistance gene candidates in bananas.

Commercial banana varieties are highly susceptible to fungal pathogens, as well as bacterial pathogens, nematodes, viruses, and insect pests. The largest known family of plant resistance genes encodes proteins with nucleotide-binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. Conserved motifs in such genes in diverse plant species offer a means for the isolation of candidate genes in banana that may be involved in plant defense. Six degenerate PCR primers were designed to target NBS and additional domains were tested on commercial banana species Musa acuminata subsp malaccensis and the Musa AAB Group propagated in vitro and plants maintained in a greenhouse. Total DNA was isolated by a modified CTAB extraction technique. Four resistance gene analogs were amplified and deposited in GenBank and assigned numbers HQ199833-HQ199836. The predicted amino acid sequences compared to the amino acid sequences of known resistance genes (MRGL1, MRGL2, MRGL3, and MRGL4) revealed significant sequence similarity. The presence of consensus domains, namely kinase-1a, kinase-2 and hydrophobic domain, provided evidence that the cloned sequences belong to the typical non-Toll/interleukin-1 receptor-like domain NBS-LRR gene family.