Search

QTLs for yield-related traits in tetraploid cotton have been widely mapped, but QTLs introduced from diploid species into tetraploid cotton background remain uninvolved. Here, a stable introgression line with the traits of small boll and seed on Chr. A12, IL197 derived from Gossypium hirsutum (2n = AADD = 52) × Gossypium arboreum (2n = AA = 26), was employed to construct the F

Environmental changes and human activities have deteriorated the quality of groundwater, which is an important source of freshwater in coastal areas. The Jiangsu Coastal Zone (JCZ), which is a typical area of the eastern China coastal zone (ECCZ), has a great demand for clean water resources due to its dense population. The groundwater in the JCZ is affected by both human activities and seawater intrusion. However, research on heavy metals in the groundwater of the JCZ is limited. This study investigated the spatial distribution characteristics and influencing factors of heavy metals in coastal groundwater of Jiangsu Province and conducted a health risk assessment (HRA). Relatively high concentrations of Cu, Cd, Pb, Co, Zn, and Ba existed in the northern JCZ, while As and B predominated in the central JCZ. The main heavy metal pollutants in the groundwater are B and As, with mean values at 0.61 mg/L and 0.02 mg/L, exceeding the standard rate reaching 48.28% and 18.07% respectively. The HRA results showed that B had the largest hazard quotient (HQ), accounting for 50.22% of the total HQs, and As was attributed to the pollutant with the largest cancer risk (CR), accounting for 99.74% of the total CRs. According to the results of the correlation analysis, heavy metals in the groundwater of JCZ mainly originated from industrial pollution, seawater intrusion, and mineral dissolution. Seawater intrusion increases the content of As and B in groundwater, leading to higher health risks. Therefore, the government should strengthen the supervision of seawater intrusion by implementing more effective water resource management policies, or adopting engineering measures such as installing subsurface physical barriers to prevent and control seawater intrusion.

Background The oncogene, malignant T-cell-amplified sequence 1 (MCTS1), has been found to be highly expressed in a variety of cancer cell lines. It has been shown to be involved in cell cycle progression and to confer a growth advantage for lymphomas and breast cancer. Nevertheless, the role of MCTS1 in contributing to the development of oral cancer remains elusive. Methods We analyzed the gene expression profiles of MCTS1 in normal oral keratinocytes and cancerous cells. Cellular proliferation, invasion, and migration experiments were performed to detect the effect of MCTS1 on the biological evolution of oral cancer. The in vitro results were verified by the in vivo lymphatic metastasis test. The underlying mechanism of MCTS1 in promoting oral cancer invasion and metastasis correlated with the epithelial-mesenchymal transition (EMT) process as revealed by western blotting. Results The results showed that MCTS1 was aberrantly expressed in oral cancer cells. MCTS1 overexpression significantly promoted tumor cell growth, proliferation, migration, and invasion. MCTS1-mediated lymphatic metastasis was verified in vivo using an intraplantar tumor model. Biomarkers associated with EMT progression were positively or negatively regulated upon knockdown or overexpression of MCTS1, respectively. Conclusions Higher MCTS1 expression in oral cancer may be connected with an unfavorable prognosis due to involvement of MCTS1. MCTS1 potentiates the growth and proliferation of oral cancer cells and subsequent metastasis by regulating cell cycle and modifying the EMT process. Keywords Oral cancer; oncogene; malignant T-cell-amplified sequence 1 (MCTS1); metastasis; invasion.

Gastric cancer is the third leading cause of cancer death worldwide. In this study, we tried to clarify the function of KLF5 in gastric cancer. Copy number variation (CNV) and the expression of KLF5 were interrogated in public datasets. The clinical significance of KLF5 amplification and gene expression in gastric cancer were evaluated. The function of KLF5 in cell proliferation was studied in gastric cancer cell lines and organoids. We found that KLF5 amplification mainly occurred in the chromosome instable tumors (CIN) and was significantly associated with TP53 mutation. In addition, higher KLF5 expression correlated with more locally invasive gastric cancer and higher T stage. Next, a KLF5 gene expression signature was curated. The genes in the signature were involved in cell development, cell cycle regulation, cell death, suggesting potential roles played by KLF5. Functional studies using siRNAs revealed that KLF5 was essential for the proliferation of gastric cancer cells. Finally, using gastric organoid models, we revealed that the proliferation of organoids was significantly inhibited after the down regulation of KLF5. Our study revealed that KLF5 was amplified and over-expressed in gastric cancer, and it may play an oncogene-like role in gastric cancer by supporting cell proliferation.

Groundwater is the primary source of water for domestic use and agricultural irrigation in Jiaodong Peninsula. This study collected 80 groundwater samples from Jiaodong Peninsula to characterize groundwater hydrogeochemical processes and the suitability of groundwater for domestic use and agricultural irrigation. The groundwater of Jiaodong Peninsula was categorized as slightly alkaline freshwater, with a Piper diagram classifying most samples as SO4·Cl-Ca·Mg and HCO3-Ca·Mg types. Major ions were Ca2+, Na+, SO42−, and HCO3−. The major processes driving the hydrochemistry of groundwater were identified as water-rock interactions as well as evaporation. The dissolution of silicate and cation exchange were the predominant hydrogeochemical processes responsible for groundwater chemistry. Four water samples showed seawater intrusion and some indicated pollution from anthropogenic activities such as industry, agriculture, and domestic sewage discharge. Overall, it was found that groundwater in most areas of Jiaodong Peninsula is suitable for domestic use and agricultural irrigation.

Hepatocellular carcinoma (HCC) is one common malignancy. The authors previously demonstrated that miR-570 regulates the development of HCC. This study detected the effect of miR-570 on cell apoptosis, angiogenesis, T cell activation, and proliferation in a tumorigenicity assay in nude mice. miR-570 mimics and negative control (NC) were transfected into SMMC7721 cells, and then, the cells were subcutaneously injected in the right flank in nude mice. Six weeks later, the dissected tumors and peripheral blood were collected. Tumor weight and volume were measured, and expression of miR-570 and apoptosis-related gene Bax/Bcl-2 was detected by quantitative real-time polymerase chain reaction. Hematoxylin and eosin, immunohistochemistry of CD31 and vascular endothelial growth factor (VEGF), TUNEL assay, and flow cytometry detection of CD4 and CD8 in peripheral blood were performed. miR-570 mimics suppressed tumor growth compared with the NC, with decreases in tumor weight and tumor volume. Very few CD31 and VEGF were found in tumor sections in miR-570 mimics group. Bax level was significantly increased, while Bcl-2 level was significantly downregulated. Significant lower ratio of CD3+CD4+ T cells and higher ratio of CD8+IFN-γ+ T cells were found in peripheral blood and tumor tissues in miR-570 mimics than NC. Collectively, miR-570 plays an important role in the proliferation, angiogenesis, and immune escape of HCC, which might be potential diagnostic and predictive biomarkers.

Several microRNAs (miRNAs) have been closely correlated with the development of hepatocellular carcinoma (HCC). However, the involvement of miR-300 in the development of HCC remains unknown. This study elucidated the potential molecular mechanisms of miR-300 in the modulation of the epithelial-mesenchymal transition (EMT) and invasion of HCC. The expression levels of miR-300 in HCC cells and clinical samples were detected by quantitative real-time PCR and in situ hybridization. The in vitro function of miR-300 in HCC was evaluated using a migration/invasion assay. Quantitative real-time PCR, western blotting, immunofluorescence and immunohistochemistry were used to validate the roles of miR-300 and FAK/PI3K/AKT in EMT progression. A dual-luciferase reporter assay was performed to confirm the target gene. miR-300 was down-regulated in HCC and significantly correlated with a poor prognosis in HCC patients. The down-regulation of miR-300 increased the invasiveness of the HCC cells, and promoted the EMT in both HCC tissues and HCC cells. In contrast, up-regulation of miR-300 led to the opposite results. Ectopic overexpression of miR-300 reversed TGF-β1-induced EMT in SMMC-7721 cells, and according to a dual-luciferase reporter assay and rescue assay, miR-300 inhibits the EMT-mediated migration and invasion of HCC cells via the targeted modulation of FAK and the downstream PI3K/AKT signaling pathway. miR-300 targeting modulates FAK, and the PI3K/AKT signaling pathway inhibits the EMT and suppresses the migration and invasion of HCC cells. Thus, miR-300 represents a promising therapeutic target for HCC.

Aims To investigate the therapeutic potential of itraconazole in oral squamous cell carcinoma (OSCC) and its molecular mechanism. Materials and methods The in vitro anti-cancer effects of itraconazole was determined by CCK-8 assay and colony formation assay. Transwell and wound healing assays were used to examine cell invasion and migration. The in vivo therapeutic efficacy of itraconazole was assessed by OSCC patient-derived xenograft (PDX) model. Western blot was performed to explore the anti-cancer mechanism. Key findings Itraconazole inhibited cell proliferation and colony formation of OSCC cells in a time and concentration dependent manner; induced cell cycle arrest and apoptosis, as well as inhibited cell invasion and migration. In the OSCC PDX model, itraconazole impeded tumor growth, reduced Ki-67 expression and induced apoptosis. Itraconazole downregulated the protein expression of Hedgehog pathway to inhibit proliferation and migration of oral squamous cell carcinoma cells, which can be revised by recombinant human sonic hedgehog protein (rSHH). Significance Itraconazole showed anti-cancer effects on OSCC via inhibiting the Hedgehog pathway.

Th17 activity has been implicated in systemic lupus erythematosus (SLE), which is a systemic autoimmune disease with a typical clinical manifestation of lupus nephritis (LN). Retinoic acid receptor-related orphan receptor gamma t (RORγt) has been shown to be important for Th17 differentiation. In this study, we evaluated the inhibition of RORγt activity by 3β-acetyloxy-oleanolic acid (AOA), a small molecule isolated from the root of Symplocos laurina, a traditional herb belonging to South China. We demonstrated that AOA can inhibit RORγt activity and prevent SLE pathogenesis in a pristane-induced LN model. The results showed that AOA decreased RORγt transcription activity in a reporter assay and prevented Th17 differentiation in vitro. In vivo studies showed that AOA treatment decreased serum anti-dsDNA antibody and alleviated renal pathologic damage as well as antibody complex accumulation in the pristane-induced LN model. These results demonstrated that AOA can improve the clinical manifestation of LN, indicating potential application in SLE therapy.

Fibre strength (FS) is an important quality attribute in the modern textile industry, which is genetically controlled by quantitative trait loci (QTLs). Fine-mapping stable QTLs for FS to identify candidate genes would be valuable for uncovering the genetic basis of fibre quality traits in cotton. Here, a single segment introgression line, IL-D2−2, from the cross of (TM-1×TX-1046) reported in our previous studies, was found to have significantly improved FS compared with the recurrent parent TM-1. To fine-map the QTLs of the FS, we further crossed IL-D2−2 with its recurrent parent TM-1 to produce F2 and F2:3 populations. QTL analysis and substitution mapping showed qFS-Chr. D02 was anchored into a 550.66 kb-interval between two markers, INTR1027 and JESPR-231. This interval contained 67 genes, among which 27 genes related to cell-wall synthesis were selected to conduct qRT-PCR. The results revealed seven genes were expressed significantly differently during the fibre secondary-wall-thickening stage (10–25 days post-anthesis), three being upregulated and four downregulated in IL-D2−2. Both GH_D02G2269 (UDP-glucosyl transferase 84B1) and GH_D02G2289 (unknown function (DUF869)) with nonsynonymous SNPs in IL-D2−2 had significantly downregulated expression, suggesting they were candidates for qFS-Chr. D02. This research provides information about marker-assisted selection for cotton fibre strength improvement.

Previous studies examining whether autophagy has a protective or deleterious role during ischemia/reperfusion (I/R) injury have reported a varying role in different organs and remains a matter of debate. The aim of the current study was to explore the role of autophagy in hindlimb I/R injury in a murine model. An increase in apoptosis was observed in vitro, in C2C12 myoblast cells, following hypoxia/reoxygenation (H/R), while downregulation of autophagic flux was induced by chloroquine as compared with the vehicle group under hypoxia and H/R conditions. In vivo, an increase in severe damage of gastrocnemius muscles was observed in the I/R and ischemia groups compared with the control group, was more severe in the I/R group compared with the ischemia group. Electron microscopy revealed an increased number of autophagosomes in the ischemia group, whereas a reduced number was detected in the I/R group. Following administration of rapamycin, the infarct size ratio and cell apoptosis was significantly reduced, while the amount of autophagosomes significantly increased in the ischemic phase. In conclusion, autophagy is upregulated in the ischemia phase and downregulated in the reperfusion phase. Notably, upregulation of autophagy via rapamycin intervention during ischemia alleviated skeletal muscle damage, suggesting a potential protective role during hindlimb I/R injury.

A recent study reported that miR-570 was the most important microRNA in the microRNA gene networks of alcoholic liver disease that has the potential of progressing to hepatocellular carcinoma. However, litter is known regarding the expression and specific function of miR-570 in the progression of hepatocellular carcinoma, especially its molecular mechanisms by which miR-570 exerts its functions and modulates the malignant phenotypes of hepatocellular carcinoma cells. Here, we observed that miR-570 was highly expressed in hepatocellular carcinoma cell lines (Bel-7404, Huh-7, and HepG2), while B7-H1 was lowly expressed, compared to nonmalignant cell line (L-02 and HL-7702). Transfection of miR-570 mimics or knockdown of B-H1 suppressed the expression of B7-H1, which promotes cell apoptosis and inhibits the cell proliferation and invasion. Using a dual-luciferase reporter system, we verified that B7-H1 is a direct target of miR-570. The overexpression of B7-H1 reversed the inhibition of proliferation and invasion by miR-570. In addition, miR-570 suppressed tumorigenicity in vivo. Hence, our observation confirmed that miR-570 works as proliferation and metastatic suppressor in hepatocellular carcinoma cells through directly targeting B7-H1 in hepatocellular carcinoma cell and rationally presents that miR-570 has the potential to be a useful clinical noninvasive diagnostics or predictive marker in human hepatocellular carcinoma.

Autoimmune lymphoproliferative syndrome (ALPS) is an incurable disease mainly caused by the defect of Fas-mediated apoptosis and characterized by nonmalignant autoimmune lymphoproliferation. Stabilizedβ-catenin could not only potentiate Fas-mediated T cell apoptosis via upregulating the expression of Fas on activated T cells, but also potentiate T cell apoptosis via intrinsic apoptotic pathway. In the present study, we introducedβ-catTgintolpr/lprmice and aimed to explore the potential role of stabilizedβ-catenin (β-catTg) in the development of ALPS-like phenotypes oflpr/lprmice. We found that the total splenocyte cells and some compositions were slightly downregulated inβ-catTglpr/lprmice, especially the CD4 and CD8 TEMcells were significantly reduced. Meanwhile, stabilizedβ-catenin obviously decreased the numbers of spleen TCRβ+CD4−CD8−T (DNT) cells, and the levels of some serum proinflammatory factors also were lowered inβ-catTglpr/lprmice. Beyond that, stabilizedβ-catenin slightly lowered the levels of the serum autoantibodies and the scores of kidney histopathology ofβ-catTglpr/lprmice compared withlpr/lprmice. Our study suggested that stabilizedβ-catenin ameliorated some ALPS-like symptoms oflpr/lprmice by potentiating Fas-independent signal-mediated T cell apoptosis, which might uncover a potential novel therapeutic direction for ALPS.

Osteosarcomas are common bone malignancies in children and adolescents. LDOC1 (leucine zipper, down-regulated in cancer 1), a tumor suppressor, is down-regulated in many cancers. In this study, we investigated the role of LDOC1 in tumor metastasis and its prognostic significance in osteosarcomas. We established osteosarcoma cells stably expressing LDOC1, driven by an HIV-based lentiviral system. We investigated the impact of LDOC1 on migration and invasion abilities in these cells using a transwell assay. LDOC1-associated changes in expression of metastasis-promoting genes were analyzed with a quantitative real-time polymerase chain reaction primer array. A xenograft tumor model (n = 7 mice/group) was used to assess the effect of LDOC1 on osteosarcoma metastasis in vivo. The overall survival and disease-free survival of osteosarcoma patients (n = 74) were analyzed retrospectively based on immunohistochemical analysis of LDOC1 levels in tumors and Kaplan–Meier analysis. LDOC1-expressing osteosarcoma cells displayed decreased migration and invasion in vitro. The quantitative real-time polymerase chain reaction primer array data showed that increased LDOC1 expression up-regulated many metastasis-suppressor genes. In the xenograft model, micro-computed tomography imaging data indicated that increased LDOC1 expression is associated with weaker lung metastasis ability. The Wnt5a signaling pathway promotes osteosarcoma metastasis; LDOC1 expression decreased Wnt5a levels in osteosarcoma cells. Kaplan–Meier analysis showed that higher LDOC1 expression was associated with improved osteosarcoma patient overall survival and disease free survival (p = 0.022). Our data show that LDOC1 is a tumor suppressor in osteosarcoma, and that it regulates metastasis of osteosarcoma cells. Furthermore, LDOC1 might be a valuable prognostic marker in osteosarcomas.

CD133-positive cancer stem cells in colon cancer are resistant to conventional chemotherapy. The aim of the present study was to investigate the effect of celecoxib, a COX-2 inhibitor, on CD133 expression in HT29 and DLD1 cells. HT29 and DLD1 cells were treated with celecoxib using different concentrations and duration. CD133 expression was detected by flow cytometry, Western blotting, immunofluorescence, and quantitative real-time PCR. Wnt signaling pathway activity was measured by luciferase assay and gene expression changes were monitored using microarray analysis. HT29 cells showed significantly decreasing levels of CD133 expression with increasing concentrations of or duration of exposure to celecoxib. CD133 mRNA relative expression in HT29 and DLD1 cells also decreased with drug exposure. Furthermore, Wnt activation in HT29 and DLD1 cells decreased with celecoxib treatment. Gene expression microarray showed stemness genes, including Lgr5, Oct4, Prominin-1, Prominin-2, CXCR4, E2F8, CDK-2, were downregulated and differentiation genes, including CEACAM5, GDF, ADFP, ICAM1, were upregulated. Our results show that CD133 expression was downregulated by celecoxib through inhibition of the Wnt signaling pathway, which may be lead to cell differentiation.

Using synthetic systems to mimic natural enzymes with high catalytic activity and distinct substrate selectivity has been a challenge for the last several decades. Hemin, the catalytic center for many protein families including cytochromes, peroxidases, myoglobins and hemoglobins, can catalyze a variety of oxidation reactions like peroxidase enzymes.[1] However, direct application of hemin as an oxidation catalyst is of significant challenge because of its molecular aggregation in aqueous solution to form catalytic inactive dimers and oxidative self-destruction in the oxidizing media, which causes passivation of its catalytic activity.[2] A potential solution to this problem is to synthetically modify the porphyrin structure to achieve a variety of iron porphyrin derivatives for improved catalytic activity or stability.[3] An alternative approach is to use high surface area materials such as zeolites, nanoparticles, silica or natural clay to support hemin to achieve improved stability or activity in epoxidation or other reactions in organic solutions.[4] For reactions in aqueous solutions, hydrogel-embedded hemin[5] or more elaborate hemin complex obtained by conjugating with specific antibodies[6] have shown activity significantly better than free molecules, which is, however, still orders of magnitude inferior to natural enzymes, not to mention the difficulties in the synthesis of such kinds of complex hemin conjugates. Therefore, the discovery and development of novel materials as supports to achieve biomimetic catalysts with enzyme-like activity is highly desired.

The aim of the present study is to explore possible role of miR-221 in the pathogenesis of HCC. Matched HCC and adjacent non-cancerous samples were assayed for the expression of miR-221 and three G1/S transition inhibitors: p27Kip1, p21WAF1/Cip1and TGF-β1 by in situ hybridization and immunohistochemistry respectively. p27Kip1 is one of miR-221’s proven targets. Real time qRT-PCR was used to investigate miR-221 and p27Kip1 transcripts in different clinical stages. Western blotting was used to analyze the expression levels of p27Kip1 protein in different clinical stages. In result, miR-221 and TGF-β1 are frequently up-regulated in HCC, while p27Kip1 and p21WAF1/Cip1 proteins are frequently down-regulated. Moreover, miR-221 and p27Kip1’s expression correlated with metastasis and miR-221’s expression also correlated with tumor size. Both of p21WAF1/Cip1and TGF-β1’s expression correlated with tumor differentiations. miR-221’s upregulation and p27Kip1’s downregulation were significantly associated with tumor stages and metastasis. In conclusion, miR-221 is important in tumorigenesis of HCC, possibly by specifically down-regulating p27Kip1, a cell-cycle inhibitor. These results indicate miR-221 as a new therapeutic target in HCC.

The presence of vector backbone sequences and selectable marker genes in transgenic plants has been the key concern for biosafety. A direct solution is to totally avoid the use of vector backbone sequences and selectable marker genes from the beginning of transgenic plant generation. In this study, the ovary-drip method was established and optimized. The key features of this method focused on the complete removal of the whole styles, and the subsequent application of a DNA solution directly to the ovaries. A vector backbone-free and selectable marker-free linear GFP cassette (Ubi-GFP -nos) was transformed into maize via the ovary-drip method. PCR analysis showed that suitable maize variety was 9818 and optimal transformation time was 18-20 h after pollination, which produced the highest PCR positive frequency (3.01%). Southern blotting analysis showed that the transgenic plants had simple integration patterns (1-2 bands). GFP transcription was de-tected by RT-PCR analysis. Green fluorescence was observed in roots and immature embryos of transgenic plants by a fluorescence microscopy.

The pollen-tube pathway is feasible to transform vector- and selectable marker-free linear gene cassettes into plants to address the biosafety issues. However, its transformation frequency is low and the screening of selectable marker-free transformants by PCR analysis is time-consuming and expensive. In this study, a linear GFP cassette (Ubi-GFP-nos) flanked by 25bp T-DNA borders was transformed into maize via the pollen-tube pathway. The forepart of each maize ear was divided into five segments (segments I-V) at an interval of two rows of kernels. The segments that were most likely to contain transgenic kernels were identified by monitoring GFP expression in the immature embryos. A total of 21 ears were transformed with the linear GFP cassette. Seven out of 19 ears exhibited positive GFP expression in the immature embryos. Transgenic kernels were primarily identified in segments III and IV. A total of 121 plants derived from kernels located within segments III and IV of the remaining two ears were screened by PCR analysis. Six plants (4.96%) showed the presence of the GFP cassette. Southern blot analysis showed that the transgenic plants had simple integration patterns. The identification of transgenic kernels would facilitate PCR screening for marker-free transgenic plants.

Although nicotine is known to protect against β-amyloid (Aβ)-induced neurotoxicity, the effect of nicotine on colchicine-induced neurotoxicity remains unknown. Colchicine is a microtubule-interfering agent and is able to induce neural apoptosis. Here we investigated whether nicotine exhibits similar neuroprotective effects and the mechanism against colchicine-induced neurotoxicity of the primarily cultured cortical neurons. In this study, we investigated the effect of nicotine on the protection of neurons against colchicine damage and evaluated the associated intracellular signaling pathways. Nicotine-induced protection was blocked by an α7 nicotinic acetylcholine receptors (nAChRs) antagonist and a phosphatidylinositol 3-kinase (PI3K) inhibitor. These results suggest that the neuroprotective effects of nicotine are mediated by the α7 nAChRs and PI3K-Akt signaling pathway. In addition, we reveal that blockade of p38 and JNK (c-Jun N-terminal kinase) signaling increased Akt signaling, thus enhancing the survival of cell treatment with colchicine. On the other hand, inhibition of constitutively active Akt enhanced p38 or JNK signaling phosphorylation. These data suggested that crosstalk between PI3K Akt and p38 or JNK signaling pathways contributed to nicotine against colchicine-induced cytotoxicity.

Adult peripheral blood-derived cells are able to differentiate into a variety of cell types, including nerve cells, liver-like cells and epithelial cells. However, their differentiation into retina-like cells is controversial. In the present study, transdifferentiation potential of human adult peripheral blood mononuclear cells into retina-like cells and integration into the retina of mice were investigated. Freshly isolated adult peripheral blood mononuclear cells were divided into two groups: cells in group I were cultured in neural stem cell medium, and cells in group II were exposed to conditioned medium from rat retinal tissue culture. After 5 days, several distinct cell morphologies were observed, including standard mononuclear, neurons with one or two axons and elongated glial-like cells. Immunohistochemical analysis of neural stem cell, neuron and retina cell markers demonstrated that cells in both groups were nestin-, MAP2 (microtubule-associated protein)- and GFAP (glial fibrillary acidic protein)-positive. Flow cytometry results suggested a significant increase in nestin-, MAP2- and CD16-positive cells in group I and nestin-, GFAP-, MAP2-, vimentin- and rhodopsin-positive cells in group II. To determine survival, migration and integration in vivo, cell suspensions (containing group I or group II cells) were injected into the vitreous or the peritoneum. Tissue specimens were obtained and immunostained 4 weeks after transplantation. We found that cells delivered by intravitreal injection integrated into the retina. Labelled cells were not detected in the retina of mice receiving differentiated cells by intraperitoneal injection, but cells (groups I and II) were detected in the liver and spleen. Our findings revealed that human adult peripheral blood mononuclear cells could be induced to transdifferentiate into neural precursor cells and retinal progenitor cells in vitro, and the differentiated peripheral blood mononuclear cells can migrate and integrate into the retina in vivo.

Insupported noble-metal catalysts the metal particles tend toaggregate during the reaction process, and the particle sizethus becomes larger which leads to lower catalytic activity.Additionally, the metal particles usually detach from thesupport when the corresponding catalyst is rubbed recipro-cally, which results in a sharp decrease of the active sites.Therefore, the design of an ideal nanostructure for supportednoble-metal catalysts that can overcome the above-men-tioned limits and, thus, display high stability, is a greatchallenge in this field.

A tissue culture-independent plant transformation method, called ovary-drip transformation, was established in which a minimal linear gene cassette [35S CaMV promoter, open reading frame of soluble modified green fluorescent protein (smGFP), and NOS terminator] was transformed into soybean. The method is characterized by directly dripping a DNA solution, which is supplemented with a surfactant, onto the ovary wound 6-8 h after self-pollination. The growth of the pollen tube was measured after self-pollination. The movement of smGFP across the passageway toward the embryo sac was monitored using fluorescein isothiocyanate-labeled DNA. The transformation frequency reached 3.2% by PCR analysis. Southern analysis of the primary transformants denoted the integration of a single site smGFP. The transgenic plants exhibited a high level of smGFP expression which was visible in the immature embryos of the transgenic soybean.

A minimal linear gene cassette (35S-phytase gene-nos) with T-DNA borders was acquired by PCR and directly introduced into soybean through the pollen tube pathway. A total of 13% of T(1 )plants were positive for phyA by specific PCR. Southern blot analyses showed that phyA insertions were harbored stably in T(2) progeny. Phytase expression level increased 2.5-fold over a 6-week period; its highest activity was 150 U/mg protein, compared to 56 U/mg protein in untransformed controls. Activity of phytase increased to 125 FTU/kg in T(3) transgenic seeds as compared to 64 FTU/kg in wild-type plants.

A novel thermostable phytase gene was cloned from Aspergillus fumigatus WY-2. It was 1459 bp in size and encoded a polypeptide of 465 amino acids. The gene was expressed in Pichia pastoris GS115 as an extracellular enzyme. The expressed enzyme was purified to homogeneity and biochemically characterized. The purified enzyme had a specific activity of 51 U/mg with an approximate molecular mass of 88 kDa. The optimum pH and temperature for activity were pH 5.5 and 55 degrees C, respectively. After incubation at 90 degrees C for 15 min, it still remained at 43.7% of the initial activity. The enzyme showed higher affinity for sodium phytate than other phosphate conjugates, and the K(m) and K(cat) for sodium phytate were 114 microM: and 102 s(-1), respectively. Incubated with pepsin at 37 degrees C for 2 h at the ratio (pepsin/phytase, wt/wt) of 0.1, it still retained 90.1% residual activity. These exceptional properties give the newly cloned enzyme good potential in animal feed applications.

s / Journal of Biotechnology 136S (2008) S236–S246 S239 Moschini, Lapan, 2006. “Grading, Minimum Quality Standards, and the Labeling of Genetically Modified Products”, Research Papers 12553, Iowa State University. Mussa, M., Rosen, S., 1978. Monopoly and product quality. Journal of Economic Theory 18, 301–317. Wilson, William W., Dahl, Bruce L., 2005. Costs and risks of testing and segregating genetically modified wheat. Review of Agricultural Economics 27 (2), 212–228. doi:10.1016/j.jbiotec.2008.07.506