Overexpression of soybean R2R3-MYB transcription factor, GmMYB12B2, and tolerance to UV radiation and salt stress in transgenic Arabidopsis.

MYB, v-myb avian myeloblastosis viral oncogene homolog, proteins play central roles in plant stress response. Previously, we identified a novel R2R3-MYB transcription factor, GmMYB12B2, which affected the expression levels of some key enzyme genes involved in flavonoid biosynthesis in transgenic Arabidopsis. In the present study, we analyzed the expression levels of GmMYB12B2 under salt, low temperature, drought, abscisic acid (ABA), and ultraviolet (UV) radiation treatments in soybean using semi-quantitative reverse transcription polymerase chain reaction. The expression of GmMYB12B2 was drastically induced by UV irradiation and salt treatment, but no response was detected under low temperature, drought, and ABA stresses. A detailed characterization of the GmMYB12B2 overexpression lines revealed that GmMYB12B2 might be involved in response of plants to UV radiation and salt stresses. Transgenic Arabidopsis lines constitutively expressing GmMYB12B2 showed an increased tolerance to salt and UV radiation treatment compared with wild-type plants. The expression levels of certain salt stress-responsive genes, such as DREB2A and RD17, were found to be elevated in the transgenic plants. These results indicate that GmMYB12B2 acts as a regulator in the plant stress response.

Expression of EpCAM and Wnt/ ß-catenin in human colon cancer.

The aims of this study were to explore the correlation between the expression of EpCAM and the Wnt/ß-catenin pathway in human colon cancer and its clinical significance for the evaluation of cancer prognosis. Samples from colon cancer, para-carcinoma, or benign intestinal tissue from individual patients (50) and from normal intestinal mucosal tissues (20) were obtained from the Pathology Department of the Shandong Province Binzhou People's Hospital (Shandong, China). Immunohistochemistry was used to detect the expression levels of EpCAM and ß-catenin proteins in these tissues, and the prognoses of the patients from whom the samples were derived were determined on follow-up examination. The corresponding in vitro mechanistic siRNA experiments were subsequently performed in the human colon cancer cell line HCT116 to observe the regulatory effects of silencing EpCAM expression on the Wnt/ß-catenin pathway. From these analyses, we determined that the expression levels of EpCAM and ß-catenin were higher in cancer tissues compared with other tissues from the same patient, and that the expression of EpCAM and Wnt/ß- catenin in colon cancers were positively correlated. The prognostic analysis showed an inverse correlation between EpCAM and Wnt/ß- catenin expression and patient prognosis. A further examination of cellular mechanisms confirmed that the silencing of EpCAM led to decreased expression of Wnt/ß-catenin, and thus reduced proliferation and increased the apoptosis ratio in the cells. These results suggest that suppression of EpCAM might be a new approach for treating colon cancer.

Meta-analysis of constitutive QTLs for disease resistance in maize and its synteny conservation in the rice genome.

We collected data regarding 340 disease resistance quantitative trait loci (QTLs) from the maize genomic database (MaizeGDB). We constructed an integrated linkage map and analyzed this map by using the BioMercator 2.1 software with IBM2 2008 Neighbors genetic linkage map as a reference. We used a meta-analysis method to identify five "consensus" synthetic resistance QTLs located on maize chromosomes 1, 3, 6, and 10, with map intervals of 5.14, 9.00, 28.50, 1.73, and 33.34 cM, respectively. The gene and marker sequences within the five "consensus" QTLs were downloaded from the MaizeGDB website. We identified eight resistance gene analogs (RGAs), through comparison of these sequences with the resistance genes of other members of Poaceae by using the online BLASTx software. On the basis of comparative mapping between the maize genetic map and the rice physical map, 54 rice and 44 maize resistance genes were projected onto the maize IBM2 2008 Neighbors genetic linkage map by using a synteny conservation approach. Additionally, 15 resistance genes in the "consensus" QTL regions were found in two "consensus" QTLs on chromosomes 3 and 6; these resistance genes included rp3, scmv2, wsm2, RG3, RG16, RG36, RG51, RG53, scmv1, mdm1, RG5, RG8, RG10, RG14, and RG29. Our results provide valuable information for fine-mapping QTL, gene cloning, and molecular breeding for resistance in maize.

Correlation between serum YKL-40 levels and albuminuria in type 2 diabetes.

We explored the correlation between serum YKL-40 levels and albuminuria in type 2 diabetes mellitus (T2DM) and its clinical significance. This study used a cross-sectional survey method. According to the American Diabetes Association 2007 Clinical Practice Recommendations, 738 patients with T2DM were divided into three groups: a normoalbuminuria group [albumin-to-creatinine ratio (ACR) <30 µg/mg, N = 360], a microalbuminuria group (ACR 30-300 µg/mg, N = 246), and a macroalbuminuria group (ACR ≥ 300 µg/mg, N = 332). The serum YKL-40 levels were determined by a quantitative sandwich enzyme-linked immunosorbent assay in all the cases and in 210 control subjects. Serum YKL-40 levels were significantly higher in the T2DM group vs the control group (P < 0.05), the macroalbuminuria group vs the microalbuminuria group (P < 0.05), and the microalbuminuria group vs the normoalbuminuria group (P < 0.05). Serum YKL-40 levels correlated with ACR in all participants. Significant correlation of YKL-40 was found with ACR, 2-h plasma glucose, glycated hemoglobin, fasting blood glucose, homeostatic model assessment of insulin resistance index, systolic blood pressure, duration, diastolic blood pressure, age, triglycerides, and high-density lipoprotein cholesterol (r-values: 0.713, 0.524, 0.515, 0.467, 0.438, 0.409, 0.407, 0.374, 0.112, 0.097, and -0.123, respectively). ACR correlated with serum YKL-40 levels (Beta = 0.555, P < 0.001). YKL-40 may be involved in the occurrence and development of diabetic nephropathy and would be useful as a new marker for the disease.

Genetic diversity of different populations and improved growth in the F1 hybrids in the swimming crab (Portunus trituberculatus).

The swimming crab, Portunus trituberculatus, is widely distributed throughout the coastal waters of Asian-Pacific nations and is an important economic species in this region. The aquaculture of swimming crabs has been plagued by problems associated with low growth rates, poor flesh quality, and weak disease resistance. To overcome these problems, selective breeding programs have been suggested as a means of genetically improving these traits in stock populations. In this study, we evaluated the genetic differentiation of 3 different geographical populations (Zhoushan: S; Laizhou Bay: L; and Haizhou Bay: H) using 8 polymorphic microsatellite loci. Nine strains of first filial generation were obtained, with 3 geographically populations as parental stock. We assessed the growth and survival rates of the F1 generation to identify new strains or breeds showing improvements in these economically important traits. Our results indicated that pairwise FST among populations was significantly higher than 0 (P = 0.0000) for every population pair, ranging from 0.0810 to 0.1083 for 3 different geographical populations. We observed significant heterosis for the growth and viability (survival) traits, although some strains (crossbred combinations) showed evidence of hybrid weakness in some growth measurements. One particular strain ("SL") outperformed other combinations, displaying the greatest extent of heterosis over the growth and viability (survival) traits. These results indicate that hybridization may be used to increase the performance of P. trituberculatus in aquaculture.

MTA1 promotes cell proliferation via DNA damage repair in epithelial ovarian cancer.

We examined whether metastasis-associated gene 1 (MTA1) promotes cell proliferation via DNA damage repair in ovarian cancer. MTA1 was successfully down-regulated using small interfering RNA in the epithelial ovarian cancer cell lines SKOV-3 and OVCAR-3. Cell growth was evaluated through MTT and colony formation assays. Fluorescence-activated cell sorting analysis was used to evaluate the distribution of cells in the cell cycle, and cytotoxicity assays were performed to study cell sensitivity to cisplatin. A neutral comet assay was used to measure levels of ionizing radiation-induced DNA damage in SKOV-3 cells, and Western blot analyses were carried out to examine the expression of key proteins involved in DNA damage repair pathways. MTA1 knockdown markedly inhibited cell growth and led to S phase cell cycle arrest. In addition, MTA1 depletion conferred sensitivity of ovarian cancer cells to cisplatin. Moreover, MTA1 depletion increased the level of ionizing radiation-induced DNA damage and caused irreparable damage, which was illustrated by a remarkable increase and persistent existence of a comet tail as well as protein expression levels of γH2AX, pRPA, and pChk1, all of which play critical roles in DNA repair. Thus, MTA1 promotes the proliferation of epithelial ovarian cancer cells by enhancing DNA repair.

Adjacent SNPs in the transcriptional regulatory region of the FADS2 gene associated with fatty acid and growth traits in chickens.

Delta-6 fatty acid desaturases are rate-limiting desaturases involved in metabolic processes of fatty acids, and they are encoded by the FADS2 gene. In the current study, an F2 resource population of Gushi chickens crossed with Anak broilers was used to investigate the genetic effects of the chicken FADS2. Two adjacent single nucleotide polymorphisms (SNPs) (g.4290C>G and g.4291C>A) were identified in the transcriptional regulatory region of the FADS2 gene by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and created restriction site-PCR-RFLP. Associations between the two SNPs with chicken fatty acid contents and growth traits were determined using linkage disequilibrium, haplotype construction, and association analysis. The two SNPs and their haplotype combinations were significantly associated with linoleic acid (C18:2), α-linolenic acid (C18:3), arachidonic acid (C20:4), body weight (BW)2, BW4, BW6, shank girth (SG)4, and breast bone length 4 (P<0.05). These results suggested that the SNPs of the FADS2 gene affected the content of essential fatty acid in muscle, and played a role in the early-stage growth rate of chickens.

Effect of MPO/H2O2/NO(-) system on nitric oxide-mediated modification of TTR amyloid and serum TTR in FAP ATTR Val30Met patients.

Amyloid deposits consist of protein fibrils and amorphous material, and this deposition is related to oxidative stress. Previously, we demonstrated the presence of high-density lipoproteins and/or lipids in amyloid deposits of familial amyloid polyneuropathy patients. In this study, the presence of myeloperoxidase (MPO) in amyloid deposits was demonstrated using immunohistochemical staining. In contrast, normal surrounding tissues were consistently negative for MPO. Nitrotyrosine was present in amyloid deposits after being exposed to the MPO/H2O2/NO(-) system by immunohistochemical staining, and the oxide mediated modification of serum transthyretin (TTR) was observed upon exposure to the MPO/H2O2 system using two-dimensional gel electrophoresis and TTR Western blotting. This observation revealed that the TTR amyloid deposits and serum TTR were oxidized by the MPO/H2O2/NO(-) system. Nitric oxide-mediated modification of TTR may play a role in amyloidogenesis in vivo.

Effects of mycophenolate mofetil on the expression of monocyte chemoattractant protein-1 and fibronectin in high glucose cultured human mesangial cells.

The effects of high glucose on the expression of monocyte chemoattractant protein-1 (MCP-1) and the main component of the extracellular matrix, fibronectin (FN), were explored in human mesangial cells (HMCs), along with the intervention effects of mycophenolate mofetil (MMF) on these indicators. Cultured HMCs were divided into five groups: 1) normal control group (5 mM glucose); 2) high glucose group (30 mM glucose); 3) mannitol osmotic pressure control group (5 mM glucose + 25 mM mannitol); 4) high glucose + MMF-10 group (30 mM glucose + 10 µg/mL MMF); 5) high glucose + MMF-100 group (30 mM glucose + 100 µg/mL MMF). At 24, 48, and 72 h, reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay methods were used to detect the effects of MMF on MCP-1 mRNA and protein and FN expression in HMCs under high glucose conditions. MCP-1 mRNA and protein expressions and FN secretion significantly increased in HMCs of the high glucose group compared with the normal control group (P < 0.01), with the highest expression observed at 48 h. MMF could reduce the MCP-1 mRNA and protein and FN expression levels (P < 0.01), and the inhibition occurred in a dose- and time-dependent manner (P < 0.05). In conclusion, MMF could inhibit MCP-1 expression and the secretion of FN, indicating that it may delay the progression of glomerulosclerosis and interstitial fibrosis in diabetic nephropathy to ultimately achieve protective effects on the kidney.

Isolation and characterization of polymorphic microsatellite markers from Coilia ectenes.

Coilia ectenes (Jordan and Seale 1905) is an important anadromous species that is an important resource at risk of extinction because of over-fishing, pollution, and coastal construction. To evaluate the genetic diversity of C. ectenes for use in breeding programs, elite microsatellite-enriched libraries were constructed and novel microsatellite markers were developed, and applied to genetically detect wild populations. Out of 92 randomly selected and sequenced clones, 89 contained a CA or GA repeat motif. Twenty-two pairs of primers were designed to investigate the polymorphism and genetic structure of a wild population collected from the Yellow River estuary, China. It was found that 2 loci were monomorphic and 20 loci were polymorphic. The number of alleles per polymorphic loci ranged from 3 to 13, with an average of 7.9. The expected heterozygosity per locus ranged from 0.05 to 0.89, with an average of 0.68. The isolated polymorphic markers are expected to be of use in future genetic breeding programs for C. ectenes, and in the assessment of genetic variation within this species.