TaCOLD1 defines a new regulator of plant height in bread wheat.

Plant height is among the most important agronomic traits that influence crop yield. However, in addition to the Rht-1 alleles, the molecular basis of plant height in bread wheat remains largely unclear. Based on wheat gene expression profiling analysis, we identify a light-regulated gene from bread wheat, designated as TaCOLD1, whose encoding protein is homologous to cold sensor COLD1 in rice. We show that TaCOLD1 protein is localized to the endoplasmic reticulum (ER) and plasma membrane. Phenotypic analyses show that overexpression of a mutated form of TaCOLD1 (M187K) in bread wheat cultivar Kenong199 (Rht-B1b) background resulted in an obvious reduction in plant height. Further, we demonstrate that the hydrophilic loop of TaCOLD1 (residues 178-296) can interact with TaGα-7A (the α subunit of heterotrimeric G protein) protein but not TaGα-1B, and the mutation (M187K) in TaCOLD1 remarkably enhances its interaction with TaGα-7A. Physical interaction analyses show that the C-terminal region of TaGα-7A, which is lacking in the TaGα-1B protein, is necessary for its interaction with TaCOLD1. Intriguingly, the C-terminal region of TaGα-7A is also physically associated with the TaDEP1 protein (an atypical Gγ subunit). Significantly, we discover that TaCOLD1 and mTaCOLD1 (M187K) can interfere with the physical association between TaGα-7A and TaDEP1. Together, this study reveals that TaCOLD1 acts as a novel regulator of plant height through interfering with the formation of heterotrimeric G protein complex in bread wheat and is a valuable target for the engineering of wheat plant architecture.

The role of plasma membrane H(+) -ATPase in jasmonate-induced ion fluxes and stomatal closure in Arabidopsis thaliana.

Methyl jasmonate (MeJA) elicits stomatal closure in many plant species. Stomatal closure is accompanied by large ion fluxes across the plasma membrane (PM). Here, we recorded the transmembrane ion fluxes of H(+) , Ca(2+) and K(+) in guard cells of wild-type (Col-0) Arabidopsis, the CORONATINE INSENSITIVE1 (COI1) mutant coi1-1 and the PM H(+) -ATPase mutants aha1-6 and aha1-7, using a non-invasive micro-test technique. We showed that MeJA induced transmembrane H(+) efflux, Ca(2+) influx and K(+) efflux across the PM of Col-0 guard cells. However, this ion transport was abolished in coi1-1 guard cells, suggesting that MeJA-induced transmembrane ion flux requires COI1. Furthermore, the H(+) efflux and Ca(2+) influx in Col-0 guard cells was impaired by vanadate pre-treatment or PM H(+) -ATPase mutation, suggesting that the rapid H(+) efflux mediated by PM H(+) -ATPases could function upstream of the Ca(2+) flux. After the rapid H(+) efflux, the Col-0 guard cells had a longer oscillation period than before MeJA treatment, indicating that the activity of the PM H(+) -ATPase was reduced. Finally, the elevation of cytosolic Ca(2+) concentration and the depolarized PM drive the efflux of K(+) from the cell, resulting in loss of turgor and closure of the stomata.

MIR156-Targeted SPL9 Is Phosphorylated by SnRK2s and Interacts With ABI5 to Enhance ABA Responses in Arabidopsis.

The miR156-targeted SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors play key roles in regulating plant development, but little is known about their function in abscisic acid (ABA) signaling. Here, we report that the miR156-targeted SPLs enhance ABA responses and contribute to the inhibition of pre-harvest sprouting. We find that SPL9 directly activates the expression of ABA responsive genes through binding to their promoters. SPL9 was further shown to physically interact with ABSCISIC ACID INSENSITIVE 5 (ABI5), a master transcription factor in ABA signaling, thus promoting its association with the promoters of ABA responsive genes. Furthermore, we reveal that the protein kinases SnRK2s interact with and phosphorylate SPL9, which is essential for its role in the activation of ABA responses. Together, our results disclose a SnRK2s-SPLs-ABI5 regulatory module in ABA signaling in Arabidopsis.

Significance of myeloid antigen expression in precursor T lymphoblastic lymphoma.

BACKGROUND AND OBJECTIVE: Precursor T lymphoblastic lymphoma (T-LBL) is a highly aggressive lymphoma. Myeloid antigen expression was found in some of the patients, and its clinical significance is worth studying. This study was to compare the clinical features, short-term efficacy and survival of T-LBL patients with or without myeloid antigen expression so as to evaluate its prognostic significance. METHODS: Forty-five T-LBL patients, with a median age of 14 years, were treated at Sun Yet-sen University Cancer Center between January 2000 and July 2008. These patients were divided into myeloid antigen-positive group (My(+) group) and myeloid antigen-negative group (My(-) group) based on the flow cytometric (FCM) analysis in bone marrow or pleural fluid. Myeloid antigen expression and its correlation with the short-term efficacy and overall survival were assessed in the two groups. RESULTS: There were 18 patients (40.0%) in the My(+) group and 27 (60.0%) in the My(-) group. The myeloid antigen expression was negatively correlated with the initial level of lactate dehydrogenase (LDH), but not with other clinical features. The remission rate was lower in the My(+) group than in the My(-) group (38.8% vs. 70.3%, P = 0.028). The 2-year overall survival rate was lower in the My(+) group than in the My(-) group (51.9% vs. 78.7%, P = 0.036). By age subgroup analysis, there were no differences in response and survival rate among children and adolescents with or without myeloid antigen expression. But the remission rate and the 2-year overall survival rate were significantly lower in adult patients with myeloid antigen expression than in patients without it. Univariate and multivariate analysis demonstrated that age and myeloid antigen expression were adverse prognostic factors. CONCLUSION: Myeloid antigen expression is a predictor of a poor response to chemotherapy, and adverse prognostic factor in adult T-LBL, but not in children with T-LBL.

Lgf-YL-9 induces apoptosis in human epidermoid carcinoma KB cells and multidrug resistant KBv200 cells via reactive oxygen species-independent mitochondrial pathway.

Pyrazolon derivatives were reported to have cytotoxicity to some tumour cells. In the present study, we investigated the effect of Lgf-YL-9 on cytotoxicity and cell apoptosis in human epidermoid carcinoma drug-sensitive parental KB cells and multidrug resistant (MDR) KBv200 cells. Lgf-YL-9 exhibited potent cytotoxicity not only to KB cells but also to KBv200 cells, and the IC(50) were 3.81 and 3.45 microg/mL in KB cells and KBv200 cells, respectively. Importantly, Lgf-YL-9 effectively inhibited tumour growth of KB cell xenografts in nude mice. Lgf-YL-9-induced cell apoptosis was confirmed by chromatin condensation, DNA fragmentation, Annexin-V and propidium iodide (PI) double-staining assay and poly(ADP-ribose) polymerase (PARP) cleavage. Furthermore, Lgf-YL-9-mediated apoptosis in KB cells and KBv200 cells was accompanied by the loss of mitochondrial membrane potential (DeltaPsi(m)), the release of cytochrome c, and the activation of caspases-3, -7, and -9, but not by intercalating to DNA. Although Lgf-YL-9-induced apoptosis was associated with the decrease of DeltaPsi(m), reactive oxygen species (ROS) reduction was interestingly observed in both cell lines. The data suggest that Lgf-YL-9 has similar cytotoxicity to drug-sensitive parental KB cells and MDR KBv200 cells. Lgf-YL-9-induced apoptosis is involved in a new ROS-independent mitochondrial dysfunction pathway, but not in intercalating to DNA.

Heterotrimeric G-proteins involved in the MeJA regulated ion flux and stomatal closure in Arabidopsis thaliana.

Heterotrimeric G-proteins play an important role in plant signalling pathways. The plant hormone methyl jasmonate (MeJA) can induce stomatal closure in many plant species. The signal cascade in MeJA-induced stomatal closure has been studied previously. However, the function of G proteins in this process has not yet been evaluated. In this study, the stomatal movement induced by MeJA in the wild-type Arabidopsis thaliana (L. Heynh.) (WS), Gα subunit loss-of-function mutant gpa1-1 and gpa1-2 guard cells were measured. Further, the transmembrane ion flux (H+, Ca2+ and K+) and reactive oxygen species (ROS) experiments were performed in guard cells from WS, GDP-ß-S pre-treated WS, gpa1-1 and gpa1-2 using non-invasive micro-test technique (NMT) and confocal technique. It was observed that the MeJA-induced stomatal closure was abolished in guard cells of gpa1 mutants. GDP-ß-S pre-treatment and gpa1 mutants impaired the MeJA-activated H+ efflux, Ca2+ influx and K+ efflux. The accumulation of ROS in gpa1-1 and gpa1-2 guard cells was also lower than that in WS guard cells under MeJA treatment. These results suggested that Gα subunits are involved in regulating the signal events in JA signal pathway and stomatal closure.

Similar metabolic changes induced by HIPVs exposure as herbivore in Ammopiptanthus mongolicus.

Herbivore-induced plant volatiles (HIPVs) are important compounds to prim neighboring undamaged plants; however, the mechanism for this priming process remains unclear. To reveal metabolic changes in plants exposed to HIPVs, metabolism of leaves and roots of Ammopiptanthus mongolicus seedlings exposed to HIPVs released from conspecific plants infested with larvae of Orgyia ericae were analyzed together with control and infested seedlings using nuclear magnetic resonance (NMR)-based metabolic technology and multi variate data analysis. Results presented showed that HIPVs exposure led to similar but specific metabolic changes compared with those induced by infestation in both leaves and roots. Furthermore, both HIPVs exposure and herbivore attack resulted in metabolic changes involving a series of primary and secondary metabolites in both leaves and roots. Taken together, these results suggested that priming of yet-damaged plants may be achieved by reconfiguring metabolic pathways in leaves and roots to make similar concentrations for all metabolites as those in seedlings infested. Therefore, we propose that improved readiness of defense induction of primed plants toward subsequent herbivore attack may be based on the similar metabolic profiling induced by HIPVs exposure as those caused by herbivore.

[Bone marrow immunophenotypes of 112 cases of lymphoid system malignant diseases].

BACKGROUND & OBJECTIVE: Diagnosis of lymphocytic leukemia and non-Hodgkin's lymphoma (NHL) is based on bone marrow morphology. Immunophenotyping will make diagnosis more precise through analyzing the origin and differentiation status of tumor, which is necessary for treatment and prognosis prediction. This study was to analyze the immunophenotypic characteristics of lymphocytic leukemia and NHL with bone marrow involvement using flow cytometry (FCM). METHODS: Bone marrow specimens from 112 patients with lymphocytic leukemia or NHL with bone marrow involvement were detected by FCM using antibodies of T, B and myeloid cell series. Using CD45/SSC gating strategy, the samples were analyzed with 5 parameters (FSC, SSC, McAb1-FITC, McAb2-PE, CD45-cytochrome). RESULTS: In 45 cases of precursor B lymphoblastic leukemia/lymphoma (B-ALL/LBL), the antigens were mainly CD19, CD10, TdT, CD34, HLA-DR, and CD20. In 32 cases of precursor T lymphoblastic leukemia/lymphoma (T-ALL/LBL), the antigens were mainly CD7, CD5, cytoplasmic (Cy)CD3, TdT, CD34, surface CD3 (sCD3), and HLA-DR. Of the 77 cases of precursor ALL/LBL, 28(36.4%) expressed myeloid-associated antigens, such as CD13 and CD33; 9 (20.0%) cases of B-ALL/LBL coexpressed CD20 and CD34; 28(87.5%) cases of T-ALL/LBL coexpressed cyCD3 and TdT. Among the 35 cases of mature B-cell malignancies, 17 cases of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) mainly expressed CD19, CD20, CD5, HLA-DR, with coexpression of CD19 and CD5; 4 cases of diffuse large B-cell lymphoma (DLBCL) mainly expressed CD19, CD20, CD10, and HLA-DR; 3 cases of Burkitt's lymphoma (BL) mainly expressed CD19, CD10, CD20, and sIgM; 1 case of mantle cell lymphoma (MCL) expressed CD5, CD19, CD20, and HLA-DR. Among the 10 mature T-cell malignancies, 5 cases of unspecialied peripheral T-cell lymphoma (PTCL) mainly expressed sCD3, CD5 and CD7, CD4 or CD8; 1 case of anaplastic large cell lymphoma (ALCL) expressed sCD3 and HLA-DR; 4 cases of NK/T-cell malignancies expressed CD56 and HLA-DR, CD4 or CD8 or CD7. Mature lymphoid system malignancies didn't express early antigens, such as CD34 and TdT, but expressed myeloid-associated antigens, especially CD13 and CD33. CONCLUSION: Multiparameter FCM can not only provide data of cell lineage and differentiation status but also detect phenotypic aberrancies, which is helpful for minimal residual disease detecting.

[Cytokine responses after lobectomy: a prospective randomized comparison of video-assisted thoracoscopic surgery and minimal incision thoracotomy].

BACKGROUND & OBJECTIVE: The cytokine network plays a pivotal role in inducing acute-phase inflammatory and immunologic responses to surgical trauma. Whether lesser release of cytokines by mini-invasive operation can reduce acute-phase responses and better preserve immune functions needs to be explored. This prospective randomized study was to compare the effects of video-assisted thoracoscopic surgery (VATS) and minimal incision thoracotomy (MIT) on serum levels of cytokines after lobectomy for clinical early stage non-small cell lung cancer (NSCLC). METHODS: From Mar. 2004 to Dec. 2006, 47 consecutive patients with early stage NSCLC (tumor size was

[Analysis of serum T-lymphocyte subsets and NK cell activity in patients with hypopharyngeal squamous cell carcinoma].

BACKGROUND & OBJECTIVE: T-lymphocyte subsets and NK cell are the major forms of cellular immunity. Study of these 2 types of cells may lead to the better understanding of the function of cellular immunity in the onset and development of carcinoma. To a certain degree, there may be cellular immunodeficiency existing in patients with hypopharyngeal squamous cell carcinoma. This study was to investigate the cellular immunity function in these patients. METHODS: T-lymphocyte subsets and NK activity were determined by flow cytometry in 78 patients with hypopharyngeal squamous cell carcinoma. Blood samples of 20 non-tumor patients were used as control. RESULTS: The levels of CD4 lymphocyte subsets, CD4/CD8 ratio, and NK activity were lower in carcinoma group than in control group, but CD8 lymphocyte level was higher in carcinoma group. The levels of CD4 lymphocyte subsets, CD4/CD8 ratio, and NK activity were lower in T3-4 group than in T1-2 group, and lower in N+ group than in N0 group. The levels of CD4 lymphocyte subsets and CD4/CD8 ratio were decreased in the carcinoma with moderate or low differentiation (P<0.05). CONCLUSIONS: T-lymphocyte subsets and NK activity are inhibited, and the cellular immunology is suppressed in the patients with hypopharyngeal squamous cell carcinoma. Analyzing T-lymphocyte subsets and NK activity would be helpful to evaluate the cellular immunologic condition of these patients.

[Reversal of multidrug resistance of tumor cells by FG020327 and its mechanism].

BACKGROUND & OBJECTIVE: Over-expression of P-glyco-protein (P-gp) in tumor cells results in multidrug resistance (MDR), and failure of chemotherapy. Combined therapy of MDR-related cytotoxins plus MDR modulators is a promising strategy to overcome clinical MDR. This study was to explore MDR reversal activity of a novel compound FG020327, and its mechanism. METHODS: MTT assay was used to evaluate MDR reversal activity of FG020327 in 2 P-gp expressing tumor cell lines, KBv200 and MCF-7/ADR. Adriamycin (ADM) accumulation in MCF-7/ADR cells was detected by fluorescence spectrophotometry. The effect of FG020327 on P-gp function was showed by rhodamine 123 (Rh123) accumulation and efflux in KBv200 cells. RESULTS: FG020327 significantly enhanced sensitivity of MDR cells to anti- tumor drugs. Five mumol/L of FG020327 enhanced sensitivity of KBv200 cells to vincristine (VCR) by 44.9 folds, the reversal activity of which was 3 times that of verapamil (VRP). However, FG020327 had little effect on drug-sensitive MCF-7 cells and KB cells. FG020327 of 2.5, 5, and 10 mumol/L also enhanced ADM accumulation in MCF-7/ADR cells by 2.3, 2.7, and 3.7 folds, respectively, but didn't affect ADM accumulation in MCF-7 cells. FG020327 enhanced Rh123 accumulation in KBv200 cells,but not in KB cells. CONCLUSIONS: FG020327 is an efficient modulator. The reversal of drug-resistance by FG020327 is probably related to enhanced anti-tumor drug accumulation, and inhibition of P-gp function in MDR tumor cells.

[The value of multiparameter flow cytometry in diagnosis of lymphocytic leukemia and bone marrow involvement of non-Hodgkin's lymphoma].

BACKGROUND & OBJECTIVE: Lymphocytic leukemia and bone marrow involvement of non-Hodgkin's lymphoma (NHL) can be diagnosed by bone marrow morphology or lymph node biopsy combined with bone marrow examination. The data of original and differentiation status of tumor can be analyzed by immunophenotype of bone marrow. These are necessary for diagnosis and treatment of lymphocytic malignancy. This study was designed to investigate the value of multiparameter flow cytometry in diagnosis of leukemia and bone marrow involvement of NHL. METHODS: The samples from 11 cases of untreated leukemia bone marrow and 41 cases of untreated NHL with bone marrow involvement and 2 cases of bone marrow whose biopsy could not be obtained due to huge mass in mediastinum and abdomen were detected by multiparameter flow cytometry using antibodies of T, B, Myeloid cell series. Three-color staining was done by CD45 combined with two cell series or special phase antibodies. Using CD45/SSC set gate to identify blast cells from mature cells. The samples were analyzed using five parameters [forward scatter (FSC), side scatter(SSC), McAb1-FITC, McAb2-PE, and CD45-cychrome]. RESULTS: Immunophenotype and diagnosis of 11 cases of leukemia were further confirmed by flow cytometry (FCM). Of 41 cases of NHL with bone marrow involvement, 33 cases (80.5%) lymph nodes immunophenotype by pathology diagnosis were consistent with bone marrow immunophenotype by FCM, 8 cases (19.5%) were inconsistent, but right diagnosis were made by combining with clinical presentation, pathology, bone marrow morphology and FCM. Another 2 cases with huge mediastinal mass and abdominal mass were diagnosed as T-NHL and B-NHL by bone marrow morphology and FCM without lymph node biopsy. CONCLUSION: Multiparameter flow cytometry of bone marrow can further ascertain the diagnosis of leukemia and NHL with bone marrow involvement. It also gives us data of cells lineage and differentiation status for leukemia and NHL. It is helpful for diagnosis, differential diagnosis, and treatment.