Upregulation of lncRNA DANCR functions as an oncogenic role in non-small lung cancer by regulating miR-214-5p/CIZ1 axis.

OBJECTIVE: Lung cancer has an unfavorable prognosis due to the lack of efficient diagnostic and therapeutic strategies. Therefore, this study sought to figure out the effect of long non-coding RNA (lncRNA) DANCR on lung cancer progression. PATIENTS AND METHODS: LncRNA DANCR and miR-214-5p expressions in non-small cell lung cancer (NSCLC) were detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Function assays, including Cell Counting Kit-8 (CCK-8) and flow cytometric analysis were conducted to clarify the role of DANCR and miR-214-5p in the progression of NSCLC. Western blot, Dual-Luciferase reporter assay, and RNA immunoprecipitation assay (RIP) were performed to elucidate the underlying mechanism. RESULTS: LncRNA DANCR was upregulated in NSCLC. The knockdown of lncRNA DANCR inhibited cell proliferation and accelerated cell apoptosis in NSCLC. LncRNA DANCR interacted with miR-214-5p. MiR-214-5p over-expression partially reversed the regulatory effects of DANCR on proliferation and apoptosis in NSCLC. In addition, CIZ1 was the downstream gene binding miR-214-5p. LncRNA DANCR could regulate the miR-214-5p/CIZ1 axis. CONCLUSIONS: Downregulation of lncRNA DANCR inhibited cell proliferation and induced cell apoptosis in NSCLC by regulating the miR-214-5p/CIZ1 axis. LncRNA DANCR may act as an oncogene and promote the progression of NSCLC.

Repairing effect of Schwann cells combined with mesenchymal stem cells on optic nerve injury in rats.

OBJECTIVE: To investigate the effect and the underlying mechanisms of combined transplantation of Schwann cells (Scs) and Mesenchymal stem cells (MSCs) on optic nerve injury in rats. MATERIALS AND METHODS: A total of 160 normal healthy adult rats were randomly divided into 4 groups: optic nerve injury group, optic nerve injury + Sc transplantation group, optic nerve injury + MSC transplantation group and optic nerve injury + Sc + MSC transplantation group. The optic nerve in the left eye of each rat was damaged via clamping to establish a model of optic nerve injury, and the right eye was used as self-control. Scs + MSCs, Scs alone, MSCs alone and normal saline were injected into the vitreous space, respectively. After the treatment, the optic nerve tissues were collected and subjected to hematoxylin-eosin (HE) staining. Next, the morphologic and pathological changes of rats in each group were observed. Retrograde labeling was utilized to count the number of retinal ganglion cells (RGCs) in the optic nerve tissues. The apoptosis of RGCs was detected using flow cytometry. Western blot was carried out to measure the protein expression level of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax). The expression and distribution of brain-derived neurotrophic factor (BDNF) and growth associated protein-43 (GAP-43) in the optic nerve of rats in each group were detected by immunohistochemistry. RESULTS: Transplantation of Scs and MSCs could maintain the morphological structures of the retina and optic nerve of rats, increase the amount of RGCs in optic nerve tissues, reduce the apoptosis of RGCs, promote the expression of the Bcl-2 protein and decrease the expression of Bax protein. In addition, our joint transplantation strategy also showed an important role in repairing optic nerve injury by clearly promoting the secretion and expression of BDNF and GAP-43, which indicated a better curative effect than that of separate application of Scs or MSCs. CONCLUSIONS: Therapy with combined use of Scs and MSCs has a significant therapeutic effect in repairing optic nerve injury.

Higher RABEX-5 mRNA predicts unfavourable survival in patients with colorectal cancer.

OBJECTIVE: The aim of the present study was to clarify the expression pattern and prognostic role of RABEX-5 mRNA in colorectal cancer (CRC) patients. PATIENTS AND METHODS: RABEX-5 mRNA levels in 187 CRC were examined by Real-time polymerase chain reaction. Then, the association of RABEX-5 mRNA levels with clinicopathological features was analyzed. Survival curves were plotted using the Kaplan-Meier method and differences in survival rates were analyzed using the log-rank test. The influence of each variable on survival was examined by the Cox multivariate regression analysis. RESULTS: RABEX-5 mRNA expression was significantly upregulated in CRC tissues compared with the adjacent noncancerous tissues (p < 0.01). By statistical analyses, high RABEX-5 mRNA expression was observed to be closely correlated with histology/differentiation (p = 0.010), N classification (p = 0.004), and TNM stage (p = 0.004). Kaplan-Meier curves showed that patients with high RABEX-5 mRNA expression showed unfavorable overall survival (OS) than the low RABEX-5 mRNA expression group (p < 0.001). Finally, univariate and multivariate analyses showed that RABEX-5 mRNA expression was an independent predictor of overall survival (p < 0.05). CONCLUSIONS: RABEX-5 mRNA may be a promising biomarker for the detection and prognosis evaluation of CRC.

Gastrodin suppresses BACE1 expression under oxidative stress condition via inhibition of the PKR/eIF2α pathway in Alzheimer's disease.

The expression of ß-site APP-cleaving enzyme 1 (BACE1) is increased in the brain of late-onset sporadic Alzheimer's disease (AD) and oxidative stress may be the potential cause of this event. The phenolic glucoside gastrodin (Gas), a main component of a Chinese herbal medicine Gastrodia elata Blume, has been demonstrated to display antioxidant activity and suppresses BACE1 expression. However, the mechanisms by which Gas suppresses BACE1 expression are not clear. Morris water maze test was performed to assess the effect of Gas treatment on memory impairments in Tg2576 mice. The level of oxidative stress in the brain of Tg2576 mice was determined by measuring the superoxide dismutase (SOD) activity, catalase (CAT) activity, and the levels of malondialdehyde (MDA) and ROS. In vivo and in vitro, we detected the expression levels of BACE1, pPKRThr446, PKR, pPERKThr981, PERK, peIF2αSer51, and eIF2α using western blot analysis. We found that Gas improved learning and memory abilities of Tg2576 transgenic mice and attenuated intracellular oxidative stress in hippocampi of Tg2576 mice. We discovered that the expression levels of BACE1, activated PKR (pPKRThr446) and activated eIF2α (peIF2αSer51) were elevated in the brains of Tg2576 mice and hydrogen peroxide (H2O2)-stimulated SH-SY5Y cells. Moreover, peptide PKR inhibitor (PRI) and Gas down-regulated BACE1 expression in Tg2576 mice and H2O2-stimulated SH-SY5Y cells by inhibiting activation of PKR and eIF2α. Gas alleviates memory deficits in mice and suppresses BACE1 expression by inhibiting the protein kinase/Eukaryotic initiation factor-2α (PKR/eIF2α) pathway. The research suggested that Gas may develop as an drug candidate in neurodegenerative diseases.

Rapid Muscle Relaxation in Siniperca chuatsi is Coordinated by Parvalbumin (PVALB) and MiR-181a.

Parvalbumins (PVALBs) are particularly abundant in the fast-contracting muscles and correlate positively with muscle relaxation speed in amphibians and fishes. MiRNAs play important roles in diverse biologic processes via binding to the 3' untranslated region (3'UTR) of the target mRNAs. In the study, four PVALB isoforms, named as PVALB1, 2, 3, and 4, were identified in the mandarin fish (Siniperca chuatsi) fast muscle and PVALB4 exhibited the highest expression level among them. By bioinformatics analysis, a putative miR-181a binding site in PVALB4 was detected and the direct interaction between miR-181a and PVALB4 was confirmed with the luciferase reporter assay. Further, when miR-181a was inhibited, it substantially increased PVALB4 mRNA expression level and the muscle relaxation rate in vivo. Taken together, the obtained results suggest that miRNA-181a/PVALB4 is an evolutionarily conserved miRNAtarget pair and their interaction is correlated with muscle relaxation rate in the mandarin fish. Therefore, the study revealed a novel molecular mechanism in the regulation of skeletal muscle relaxation in fish.

Association of methylenetetrahydrofolate reductase (MTHFR) gene polymorphism with ischemic stroke in the Eastern Chinese Han population.

The association between the MTHFR genetic polymorphism and ischemic stroke has been reported by a number of investigators. However, the results have been controversial and conflicting. The aim of this study was to explore the association between the MTHFR variants C677T and A1298C and the risk of ischemic stroke in an Eastern Chinese Han population. A total of 199 patients with ischemic stroke and 241 controls were recruited. Genotyping of the MTHFR C677T and A1298C polymorphisms was carried out using the Taqman 7900HT Sequence Detection System. The overall estimates (odds ratio: OR) for the allele (C) and genotype (AC+CC) of the A1298C polymorphism were 1.57 [95% confidence interval (CI) = 1.16-2.10], and 2.36 (95%CI = 1.39-4.00), respectively, establishing significant association of the MTHFR A1298C polymorphism with ischemic stroke. In contrast, there were no statistically significant differences compared to controls between MTHFR C677T polymorphic variants in the association ischemic stroke risk. Furthermore, haplotype-based analysis demonstrated that compared with the C-677-A-1298 haplotype, the C-677-C-1298 and T-677-C-1298 haplotypes showed significant increased risk of ischemic stroke (OR = 1.56; 95%CI = 1.07- 2.2; P = 0.02; OR = 1.76; 95%CI = 1.17-2.65; P < 0.01, respectively). We concluded that the A1298C polymorphism and the haplotypes C-677-C-1298 and T-677-C-1298 in MTHFR might modulate the risk of ischemic stroke in the Eastern Chinese Han population.

miR-143: a novel regulator of MyoD expression in fast and slow muscles of Siniperca chuatsi.

The fast and slow skeletal muscle fibers in fish are separated to a much greater degree than in mammals. MyoD is required for the maintenance of normal fiber type balance in muscles. So far, the upstream regulatory factors of MyoD in terms of controlling muscle phenotype are poorly understood. In the present study, we used Siniperca chuatsi as a model system and demonstrated that miR-143 expression was negatively correlated with MyoD expression in the fast and slow muscles of S. chuatsi. The luciferase reporter assay further verified the direct interaction between the miR-143 and MyoD. The miR-143 suppression also led to the significant increase in MyoD and fast myosin heavy chain gene expression in vivo. Taken together, our studies indicated that miRNA participates in controlling the performance of different muscle fiber types in vertebrates.

Differential activity of GSK-3 isoforms regulates NF-κB and TRAIL- or TNFα induced apoptosis in pancreatic cancer cells.

While TRAIL is a promising anticancer agent due to its ability to selectively induce apoptosis in neoplastic cells, many tumors, including pancreatic ductal adenocarcinoma (PDA), display intrinsic resistance, highlighting the need for TRAIL-sensitizing agents. Here we report that TRAIL-induced apoptosis in PDA cell lines is enhanced by pharmacological inhibition of glycogen synthase kinase-3 (GSK-3) or by shRNA-mediated depletion of either GSK-3α or GSK-3ß. In contrast, depletion of GSK-3ß, but not GSK-3α, sensitized PDA cell lines to TNFα-induced cell death. Further experiments demonstrated that TNFα-stimulated IκBα phosphorylation and degradation as well as p65 nuclear translocation were normal in GSK-3ß-deficient MEFs. Nonetheless, inhibition of GSK-3ß function in MEFs or PDA cell lines impaired the expression of the NF-κB target genes Bcl-xL and cIAP2, but not IκBα. Significantly, the expression of Bcl-xL and cIAP2 could be reestablished by expression of GSK-3ß targeted to the nucleus but not GSK-3ß targeted to the cytoplasm, suggesting that GSK-3ß regulates NF-κB function within the nucleus. Consistent with this notion, chromatin immunoprecipitation demonstrated that GSK-3 inhibition resulted in either decreased p65 binding to the promoter of BIR3, which encodes cIAP2, or increased p50 binding as well as recruitment of SIRT1 and HDAC3 to the promoter of BCL2L1, which encodes Bcl-xL. Importantly, depletion of Bcl-xL but not cIAP2, mimicked the sensitizing effect of GSK-3 inhibition on TRAIL-induced apoptosis, whereas Bcl-xL overexpression ameliorated the sensitization by GSK-3 inhibition. These results not only suggest that GSK-3ß overexpression and nuclear localization contribute to TNFα and TRAIL resistance via anti-apoptotic NF-κB genes such as Bcl-xL, but also provide a rationale for further exploration of GSK-3 inhibitors combined with TRAIL for the treatment of PDA.

SOX2 promotes dedifferentiation and imparts stem cell-like features to pancreatic cancer cells.

SOX2 (Sex-determining region Y (SRY)-Box2) has important functions during embryonic development and is involved in cancer stem cell (CSC) maintenance, in which it impairs cell growth and tumorigenicity. However, the function of SOX2 in pancreatic cancer cells is unclear. The objective of this study was to analyze SOX2 expression in human pancreatic tumors and determine the role of SOX2 in pancreatic cancer cells regulating CSC properties. In this report, we show that SOX2 is not expressed in normal pancreatic acinar or ductal cells. However, ectopic expression of SOX2 is observed in 19.3% of human pancreatic tumors. SOX2 knockdown in pancreatic cancer cells results in cell growth inhibition via cell cycle arrest associated with p21(Cip1) and p27(Kip1) induction, whereas SOX2 overexpression promotes S-phase entry and cell proliferation associated with cyclin D3 induction. SOX2 expression is associated with increased levels of the pancreatic CSC markers ALDH1, ESA and CD44. Importantly, we show that SOX2 is enriched in the ESA(+)/CD44(+) CSC population from two different patient samples. Moreover, we show that SOX2 directly binds to the Snail, Slug and Twist promoters, leading to a loss of E-Cadherin and ZO-1 expression. Taken together, our findings show that SOX2 is aberrantly expressed in pancreatic cancer and contributes to cell proliferation and stemness/dedifferentiation through the regulation of a set of genes controlling G1/S transition and epithelial-to-mesenchymal transition (EMT) phenotype, suggesting that targeting SOX2-positive cancer cells could be a promising therapeutic strategy.

Systematic identification and differential expression profiling of MicroRNAs from white and red muscles of siniperca chuatsi.

MicroRNAs (miRNAs) participate in the regulation of myogenesis and muscle physiological function. Most skeletal muscles in vertebrates contain a mixture of fibertypes. So far, the regulatory mechanism of the miRNA in terms of controlling muscle phenotype is poorly understood. In the present study, we use Siniperca chuatsi as a model system and demonstrate that miRNAs are involved in regulating the physiological processes and metabolism of different muscle fibers in vertebrates. The miRNA transcriptomes of the white muscle, red muscle, and five other tissues from Siniperca chuatsi were profiled using Solexa deep sequencing. We characterized 186 conserved miRNAs and 3 novel miRNAs from the two small RNA libraries of white and red muscles. Among the 155 miRNAs overlapped between the two libraries, we identified 60 significantly expressed miRNAs between the two types of muscle fibers. Using integrative miRNA target-prediction and network-analysis approaches, an interaction network of differentially expressed and muscle-related miRNAs and their putative targets were constructed. Sch-miR-181a-5p that could act to control the performance of the different muscle fiber types by targeting the myostatin gene was identified.

320-detector CT coronary angiography with prospective and retrospective electrocardiogram gating in a single heartbeat: comparison of image quality and radiation dose.

OBJECTIVES: To compare the image quality, radiation dose and diagnostic accuracy of 320-detector CT coronary angiography with prospective and retrospective electrocardiogram (ECG) gating in a single heartbeat. METHODS: Two independent reviewers separately scored image quality of coronary artery segment for 480 cardiac CT studies in a prospective group and a retrospective group (240 patients with a heart rate <65 beats per minute in each group). The two groups matched well for clinical characteristics and CT parameters. There was good agreement for image quality scores of coronary artery segment between the independent reviewers (κ=0.73). Of the 7023 coronary artery segments, the image quality scores of the prospective group and retrospective group were not significantly different (p>0.05). The mean radiation dose was 10.0 ± 3.5 mSv (range 6.2-21.6 mSv) for prospective ECG gating at 65-85% of R-R interval (the interval between the R-wave of one heartbeat to the R-wave of the next). The mean radiation dose for retrospective ECG-triggered modulated scans was 23.2 ± 3.4 mSv (range 17-27.4 mSv). The mean radiation dose was 57% lower for prospective gating than for retrospective gating (p<0.01). RESULTS: Compared with coronary angiography, the results for prospective vs retrospective ECG gating were 92% vs 90% for sensitivity (p=0.23), 89% vs 91% for specificity (p=0.19), 90% vs 93% for positive predictive value (p=0.25) and 92% vs 95% for negative predictive value (p=0.21) for lesions with ≥50% stenosis, respectively. CONCLUSION: 320-detector CT coronary angiography performed with prospective ECG gating has similar subjective image quality scores, but a 57% lower radiation dose than retrospective ECG gating in a single heartbeat.

Molecular characterization and structure analysis of RPL10/QM-like protein from the red drum Sciaenops ocellatus (Sciaenidae).

The QM-like gene encodes a ribosomal protein L10. Besides housekeeping roles in protein synthesis, QM-like proteins have multiple extraribosomal functions during cell growth, cell differentiation and apoptosis. We obtained the full-length cDNA of QM-like protein (designated as SoQM) from the salt water game fish Sciaenops ocellatus, using RACE-PCR. The sequence consists of 740 bp, encoding 215-amino acid residues with 24.60 kDa. The AA sequence of the SoQM protein contains a series of functional motifs that belong to the QM family signature, which is conserved among different species. The SoQM gene contains five introns and six exons. The expression pattern of SoQM as determined by RT-PCR indicated that SoQM mRNA was expressed in all tissues tested, including brain, gill, head-kidney, intestine, stomach, heart, spleen, blood, muscle, and gonads. The phylogenetic tree constructed with MEGA 4.0 showed that SoQM clusters together with that of other fish. It was found that the sequences of the SoQM gene are highly conserved, suggesting the fundamental and critical functions of SoQM in S. ocellatus. The three-dimensional structure of the SoQM protein core domain (4~169) was predicted by the Swiss-Model program. Compared with QM proteins in other species, the main structure of SoQM protein was conserved, while the C-terminal domain was different from other QM-like proteins. Prediction of the three-dimensional structure of SoQM would provide valuable insight into the molecular basis of protein function, allowing an effective design of experiments, such as site-directed mutagenesis, studies of disease-related mutations or structure-based design of specific inhibitors.

Mutant K-Ras increases GSK-3ß gene expression via an ETS-p300 transcriptional complex in pancreatic cancer.

Glycogen synthase kinase-3 beta (GSK-3ß) is overexpressed in a number of human malignancies and has been shown to contribute to tumor cell proliferation and survival. Although regulation of GSK-3ß activity has been extensively studied, the mechanisms governing GSK-3ß gene expression are still unknown. Using pancreatic cancer as a model, we find that constitutively active Ras signaling increases GSK-3ß gene expression via the canonical mitogen-activated protein kinase signaling pathway. Analysis of the mechanism revealed that K-Ras regulates the expression of this kinase through two highly conserved E-twenty six (ETS) binding elements within the proximal region. Furthermore, we demonstrate that mutant K-Ras enhances ETS2 loading onto the promoter, and ETS requires its transcriptional activity to increase GSK-3ß gene transcription in pancreatic cancer cells. Lastly, we show that ETS2 cooperates with p300 histone acetyltransferase to remodel chromatin and promote GSK-3ß expression. Taken together, these results provide a general mechanism for increased expression of GSK-3ß in pancreatic cancer and perhaps other cancers, where Ras signaling is deregulated.

The unique localization of ZFP185 at uropod of mouse T lymphocytes.

Zinc finger protein 185 (ZFP185), which contains a conservative Lin-l 1, Isl-1 and Mec-3 (LIM) domain at its C-terminus, is a member of mouse LIM protein family. Although brahma-related gene 1(BRG1), the upstream regulation gene of ZFP185, is essential in the development and activation of T lymphocytes, there is no report on the involvement of ZFP185 in lymphocyte activation. The expression of ZFP185 protein was up-regulated significantly in T lymphocytes during their activation by phorbol 12-myristate 13-acetate (PMA)/ionomycin, compared with non-activated T lymphocytes, as determined by western blot assays. However, over-expression of ZFP185 in EL4 cells showed no significant effect on the cell proliferation. Importantly, expression of ZFP185-enhanced green fluorescent protein fusion protein in EL4 cells showed that ZFP185 uniquely localized in the uropod of lymphocytes as observed with a two-photon laser scanning confocal microscope. The direct interaction between ZFP185 and actin was supported by co-immunoprecipitation and immunofluorescence assays. The special distribution of ZFP185 in uropod indicates the potential involvement of ZFP185 in T-cell morphology and movement.

ZNF185, an actin-cytoskeleton-associated growth inhibitory LIM protein in prostate cancer.

We have recently identified ZNF185 as a gene that is downregulated in prostate cancer (PCa), in part via epigenetic alteration, and maybe associated with disease progression. In this study, we cloned the ZNF185 cDNA from normal human prostate tissues and investigated its biological function. We show that ZNF185 is a novel actin-cytoskeleton-associated Lin-l 1, Isl-1 and Mec-3 (LIM) domain-containing protein that localizes to F-actin structures, and is enriched at focal adhesions. We find that the NH(2)-terminal region, which we designate the actin-targeting domain, facilitates ZNF185 binding to actin in vitro and is both necessary and sufficient to mediate actin-cytoskeleton targeting of ZNF185, whereas the LIM domain, which is localized in the COOH-terminus is dispensable for this phenomenon. Interestingly, ectopic expression of full-length ZNF185, but not a mutant lacking the actin-targeting domain, could suppress proliferation and anchorage-independent growth of PCa cells. Together, our data suggest that ZNF185 may function as a tumor-suppressor protein by associating with the actin-cytoskeleton.

QTL mapping of ten agronomic traits on the soybean ( Glycine max L. Merr.) genetic map and their association with EST markers.

A set of 184 recombinant inbred lines (RILs) derived from soybean vars. Kefeng No.1 x Nannong 1138-2 was used to construct a genetic linkage map. The two parents exhibit contrasting characteristics for most of the traits that were mapped. Using restricted fragment length polymorphisms (RFLPs), simple sequence repeats (SSRs) and expressed sequence tags (ESTs), we mapped 452 markers onto 21 linkage groups and covered 3595.9 cM of the soybean genome. All of the linkage groups except linkage group F were consistent with those of the consensus map of Cregan et al. (1999). Linkage group F was divided into two linkage groups, F1 and F2. The map consisted of 189 RFLPs, 219 SSRs, 40 ESTs, three R gene loci and one phenotype marker. Ten agronomic traits-days to flowering, days to maturity, plant height, number of nodes on main stem, lodging, number of pods per node, protein content, oil content, 100-seed weight, and plot yield-were studied. Using winqtlcart, we detected 63 quantitative trait loci (QTLs) that had LOD>3 for nine of the agronomic traits (only exception being seed oil content) and mapped these on 12 linkage groups. Most of the QTLs were clustered, especially on groups B1 and C2. Some QTLs were mapped to the same loci. This pleiotropism was common for most of the QTLs, and one QTL could influence at most five traits. Seven EST markers were found to be linked closely with or located at the same loci as the QTLs. EST marker GmKF059a, encoding a repressor protein and mapped on group C2, accounted for about 20% of the total variation of days to flowering, plant height, lodging and nodes on the main stem, respectively.

A salt-responsive receptor-like kinase gene regulated by the ethylene signaling pathway encodes a plasma membrane serine/threonine kinase.

NTHK1 is a salt-inducible ethylene receptor gene in tobacco. Transgenic tobacco plants for this gene show reduced ethylene sensitivity. Using cDNA microarray analysis, we were able to identify those genes that have different expression levels between NTHK1 transgenic plants and wild-type plants under salt stress conditions. One of these, AtLecRK2, which encodes a receptor-like kinase with an extracellular lectin-like domain, was characterized in detail in the present study. AtLecRK2 contains a signal peptide, an extracellular lectin-like domain, a single transmembrane domain and a cytoplasmic protein kinase domain. AtLecRK2 is transcribed in the root, flower and leaf but not in the stem. In wild-type Arabidopsis, salt stress induced the transcription level of AtLecRK2, whereas in the transgenic NTHK1 Arabidopsis induction of the AtLecRK2 transcript was inhibited and retarded. AtLecRK2 was constitutively overexpressed in the ethylene-overproducer mutant, eto1-1, and could be induced by ethylene. However, in the ethylene-insensitive mutant, ein2-1, the salt-induced expression pattern of AtLecRK2 was the same as that in wild-type plants. The results demonstrate that the induction of AtLecRK2 in response to salt stress is regulated by the ethylene signaling pathway. The induction was inhibited by the ethylene receptor, NTHK1, while it was independent of EIN2. The kinase activity of AtLecRK2 was also studied. We found that that AtLecRK2 can be autophosphorylated and has serine/threonine kinase activities. The subcellular localization of AtLecRK2-GFP in onion epidermal cells indicates that AtLecRK2 is localized on the plasma membrane.

Chronic morphine treatment and withdrawal induce up-regulation of c-Jun N-terminal kinase 3 gene expression in rat brain.

Chronic opiate applications produce long-term impacts on many functions of the brain and induce tolerance, dependence, and addiction. It has been demonstrated that opioid drugs are capable to induce apoptosis of neuronal cells, but the mechanism is not clear. c-Jun N-terminal kinase 3 (JNK3), specifically expressed in brain, has been proved to mediate neuronal apoptosis and is involved in opiate-induced cell apoptosis in vitro. The present study investigated the effect of opioid administration on expression of JNK3, an important mediator involved in apoptosis of neurons, in rat brain. Our results showed that single or chronic injection of morphine resulted in a 45-50% increase in the level of JNK3 mRNA in frontal cortex, while no significant change was detected in other brain regions such as thalamus, hippocampus and locus coeruleus. Similar to what was observed after the acute or chronic morphine administration, no significant change in JNK3 expression was detected in locus coeruleus following cessation of the chronic morphine administration. However, interestingly, sustained elevation of JNK3 expression peaked on day 14 after cessation of morphine treatment was observed in the brain regions such as hippocampus and thalamus, where acute or chronic morphine treatment did not cause any significant change in JNK3 gene expression. The increased JNK3 mRNA in these brain areas returned to the control levels in 28 days following cessation of chronic morphine treatment. Taken together, these results demonstrated for the first time that the expression of JNK3 gene is regulated by opioids and that chronic opioid administration and withdrawal could induce sustained elevation of JNK3 mRNA in many important brain areas. The changes in JNK3 gene expression in brain induced by chronic opioid treatment may play a role in opioid-induced apoptosis and neurotoxicity.

An EREBP/AP2-type protein in Triticum aestivum was a DRE-binding transcription factor induced by cold, dehydration and ABA stress.

We characterize one transcription factor of DRE-binding proteins (TaDREB1) that was isolated from a drought-induced cDNA library of wheat (Triticum aestivum L.). TaDREB1 contains one conserved EREBP/AP2 domain, and shows similarity with Arabidopsis thaliana DREB family members in both overall amino-acid sequences and the secondary structure arrangement within the DNA-binding motifs. In yeast one-hybrid system, TaDREB1, can specially activate the genes fused with the promoter containing three tandemly repeated copies of the wild-type DRE sequence: TACCGACAT. In different wheat cultivars, the Ta DREB1 gene is induced by low temperature, salinity and drought; and the expression of Wcs120 that contains DRE motifs in its promoter is closely related to the expression of TaDREB1. These results suggest that TaDREB1 functions as a DRE-binding transcription factor in wheat. We also observed the dwarf phenotype in transgenic rice (T0) overexpressing TaDREB1.

A soybean gene encoding a proline-rich protein is regulated by salicylic acid, an endogenous circadian rhythm and by various stresses.

A cDNA clone of a soybean gene encoding a proline-rich protein (PRP) was characterized and designated SbPRP ( Soy bean Proline- rich Protein). The SbPRP protein is a putative bimodular protein of 126 amino acids with a proline-rich domain and a hydrophobic cysteine-rich domain plus a signal peptide at the N terminal. Southern analysis indicates the presence of a single copy of the SbPRP gene in the soybean genome. The SbPRP gene expression was investigated and the results demonstrate that it accumulates in leaves and epicotyls of soybean seedlings, but not in cotyledons, hypocotyls and roots. The SbPRP mRNA was also expressed in response to salicylic acid and virus infection. In addition, the SbPRP gene transcription was regulated by circadian rhythm, salt stress, drought stress and plant hormones. These results indicate that the SbPRP gene might play a role in plant responses to multiple internal and external factors.