Genomic characterization reveals distinct mutational landscapes and therapeutic implications between different molecular subtypes of triple-negative breast cancer.

Triple-negative breast cancer (TNBC) has high heterogeneity, poor prognosis, and limited treatment success. Recently, an immunohistochemistry-based surrogate classification for the "Fudan University Shanghai Cancer Center (FUSCC) subtyping" has been developed and is considered more suitable for clinical application. Seventy-one paraffin-embedded sections of surgically resected TNBC were classified into four molecular subtypes using the IHC-based surrogate classification. Genomic analysis was performed by targeted next-generation sequencing and the specificity of the subtypes was explored by bioinformatics, including survival analysis, multivariate Cox regression, pathway enrichment, Pyclone analysis, mutational signature analysis and PHIAL analysis. AKT1 and BRCA1 mutations were identified as independent prognostic factors in TNBC. TNBC molecular subtypes encompass distinct genomic landscapes that show specific heterogeneities. The luminal androgen receptor (LAR) subtype was associated with mutations in PIK3CA and PI3K pathways, which are potentially sensitive to PI3K pathway inhibitors. The basal-like immune-suppressed (BLIS) subtype was characterized by high genomic instability and the specific possession of signature 19 while patients in the immunomodulatory (IM) subtype belonged to the PD-L1 ≥ 1% subgroup with enrichment in Notch signaling, suggesting a possible benefit of immune checkpoint inhibitors and Notch inhibitors. Moreover, mesenchymal-like (MES) tumors displayed enrichment in the receptor tyrosine kinase (RTK)-RAS pathway and potential sensitivity to RTK pathway inhibitors. The findings suggest potential treatment targets and prognostic factors, indicating the possibility of TNBC stratified therapy in the future.

[Effect and mechanism of acupoint injection on influenza A virus induced pneumonia in mice].

OBJECTIVE: To investigate the effect and mechanism of acupoint injection with 0.1% vitamin C+vitamin B complex solution (VC+VBCo) at "Tiantu" (CV 22), "Quchi" (LI 11) and "Zusanli" (ST 36) in mouse model of pneumonia induced by influenza A virus (A/PR/8/34 [H1N1], PR8). METHODS: Sixty male ICR mice were randomized into 6 groups, i.e. control group, model group, acupoint injection group, intraperitoneal injection group, non-target point group and ribavirin group, 10 mice in each one. Except the control group, the pneumonia models were induced by slow nasal dripping PR8 virus in the other groups. On the 2nd day of experiment, VC+VBCo solution, 40 µL was injected at "Tiantu" (CV 22), "Quchi" (LI 11, left) and "Zusanli" (ST 36, left) in the acupoint injection group; VC+VBCo solution, 120 µL was injected intraperitoneally in the intraperitoneal injection group; VC+VBCo solution, 40 µL was injected at non-target acupoints (0.5 cm away from "Tiantu" [CV 22] to the left side, "Quchi" [LI 11, left] and "Zusanli" [ST 36, left]) in the non-target point group; and ribavirin solution, 120 µL was injected intraperitoneally in the ribavirin group. The intervention was delivered once daily, for consecutive 7 days. Three parallel experiments were undertaken. The mean death rate and survival time were assessed in each group, the body mass and lung index were compared among groups. Using HE staining, the morphology of lung tissue was observed; and with real-time fluorescence quantitative PCR, viral load in lung tissue was detected. The concentrations of inflammatory factors (tumor necrosis factor α [TNF-α], interleukin [IL]-1ß, IL-10) were detected in lung tissue of each group using ELISA; and those of oxidative stress markers (superoxide dismutase [SOD], glutathione peroxidase [GSH-Px], malondialdehyde [MDA]) were detected with chemiluminescence method. RESULTS: Compared with the control group, the body mass was decreased and lung index was increased in the model group (P<0.01). In comparison with the model group, body mass was increased in the acupoint injection group (P<0.05), lung index was reduced in the acupoint injection group the and ribavirin group (P<0.05); the mean death rate was decreased and the mean survival time prolonged in the mice of the acupoint injection group (P<0.01, P<0.05); and the mean death rate was reduced in the mice of the ribavirin group (P<0.05). In the model group, the alveolar structure was not integral, the alveolar septum was thickened, inflammatory cells were infiltrated and red blood cells exudated seriously (P<0.01). Compared with the model group, in the acupoint injection group and the ribavirin group, the alveolar structure was integral, the thickened alveolar septum was alleviated; and the infiltration of inflammatory cells and the exudation of red blood cells were reduced remarkably. The viral load was reduced in the mice of the ribavirin group when compared with the model group (P<0.01). Compared with the control group, the concentrations of TNF-α, IL-1ß and MDA in lung tissue were increased and those of IL-10, SOD and GSH-Px were reduced in the model group (P<0.01). In the acupoint injection group and the ribavirin group, the concentrations of TNF-α, IL-1ß and MDA were reduced in lung tissue and those of IL-10, SOD and GSH-Px were increased (P<0.05, P<0.01) when compared with the model group. CONCLUSION: Acupoint injection with VC+VBCo solution may alleviate inflammatory responses and oxidative stress in lung tissue of the PR8-induced pneumonia mice, improve survival rate and prolong the survival time in the case of no effect of the viral load.

Empirical validation of the psychological concept of a perceived feeling of 'energy': Advancement into the study of positive psychology.

The paradigm of positive psychology, significant in nature, helps to explain the proactivity and motivation of human agency, such as a secondary school student's state of autonomy, confidence, and personal resolve to strive for optimal learning and/or non-learning experiences. Our recent research development, in tandem with other scholars' inquiries, has focused on one aspect of positive psychology-namely, a person's achievement of 'optimal best', which reflects the maximization of his/her state of functioning (e.g., cognitive functioning). Capitalizing on our previous research, we develop a psychological concept that we term as a 'perceived feeling of energy'. A perceived feeling of energy (e.g., a perceived feeling of liveliness) is proposed to act as a 'motivational engine', or as a central driver, which then could predict and enhance a person's achievement of optimal best. Six hundred and twenty-seven university students (N = 438 women, 189 men) responded to a suite of self-report questionnaires. Structural equation modelling (SEM) techniques were used to test a conceptual model, where we focused on the antecedent (i.e., the direct impact of self-efficacy on a perceived feeling of energy) and consequence of a perceived feeling energy (i.e., the impact of a perceived feeling of energy on personal resolve, and the sustaining of optimal best). Analysis of results showed support for our original hypothesized model-for example: self-efficacy as an antecedent of energy and the central role of the energy as a predictor and potential mediator of future outcomes.

Biliary stent combined with iodine-125 seed strand implantation in malignant obstructive jaundice.

BACKGROUND: Malignant obstructive jaundice is mainly caused by cholangiocarcinoma. Only a few patients are indicated for surgical resection, and the 3-year survival rate is < 50%. For patients who are not eligible for surgery, biliary stent placement can relieve biliary obstruction and improve liver function and quality of life. However, restenosis after biliary stents has a poor prognosis and is a clinical challenge. Biliary stent combined with iodine-125 (125I) seed implantation can prolong stent patency and improve survival. AIM: To evaluate the safety and efficacy of biliary stent combined with 125I seed strand implantation in malignant obstructive jaundice. METHODS: We enrolled 67 patients between January 2016 and June 2018 with malignant obstructive jaundice and randomized them into a biliary stent combined with 125I seed strand treatment (combined) group (n = 32) and biliary stent (control) group (n = 35). All patients underwent enhanced computed tomography and magnetic resonance imaging and were tested for biochemical and cancer markers. Twelve patients underwent pathological examination before surgery. All patients were followed up by telephone or clinical visit. Postoperative liver function improvement, postoperative complications, stent patency time, and survival time were compared between the two groups. Prognostic risk factors were evaluated. RESULTS: Technical success was achieved in all patients in both groups. Postoperative liver function improved significantly in all patients (total bilirubin, direct bilirubin, alanine aminotransferase, and aspartate aminotransferase decreased significantly in all patients, the P values were less than 0.05). There was no significant difference in preoperative or postoperative indexes between the two groups for changes in total bilirubin (P = 0.147), direct bilirubin (P = 0.448), alanine aminotransferase (P = 0.120), and aspartate aminotransferase (P = 0.387) between the two groups. The median stent patency time of the combined group was 9.0 ± 1.4 mo [95% confidence interval (CI): 6.3-11.8 mo], which was significantly longer than the that of the control group (6.0 ± 0.3 mo, 95%CI: 5.5-6.5 mo, P = 0.000). The median survival time of the combined group was 11.0 ± 1.4 mo (95%CI: 8.2-13.7 mo), which was significantly longer than that of the control group (7.0 ± 0.3 mo, 95%CI: 6.4-7.6 mo, P = 0.000). Location of obstruction and number of stents were independent risk factors affecting prognosis. CONCLUSION: Biliary stent combined with 125I seed strand implantation is safe and effective in malignant obstructive jaundice and improves stent patency time and median survival time.

Clinical-Pathologic Analysis of Breast Cancer With PIK3CA Mutations in Chinese Women.

PURPOSE: Mutations of PIK3CA have recently been shown to play an important role in the pathogenesis and progression of breast neoplasms. The prevalence of PIK3CA in Chinese breast cancer patients may be underestimated. Therefore, we investigated the distribution of somatic PIK3CA mutation in Chinese breast cancer patients and explored their role in tumor phenotypes. METHODS: Mutational analysis of PIK3CA was done in 113 primary breast cancers of Chinese women used Amplification refractory mutation system (ARMS). The relationship of PIK3CA mutations with several clinicopathologic characteristics was analyzed. RESULTS: PIK3CA gene mutation was identified in 43(38.05%) cases and has a more significant difference between exon 9 and 20. HER2 gene amplification was 32.6% in 43 cases of PIK3CA mutation, but 37.1% in 70 cases of non-mutation (χ2 = 0.245, P > 0.05). There was no significant correlation of the age distribution, lymph node status, histological tumor grading, ER and/or PR and P53 between 2 groups (P > 0.05). CONCLUSION: A high frequency of somatic PIK3CA mutation was detected in Chinese breast cancer patients, especially in exon 20. The relationship between PIK3CA gene mutation and clinical pathological features of breast cancer needs to be further studied in a large series of patients. PIK3CA mutations seem to have the potential to be used in target treatment and as an indicator of prognosis.

Advancing the Study of Positive Psychology: The Use of a Multifaceted Structure of Mindfulness for Development.

Positive psychology, as a distinctive paradigm, focuses on the remedy of pathologies and, by contrast, the promotion of positive experiences and conditions in life (e.g., encouraging a state of flourishing). Positive psychology, in its simplistic form, may provide evidence and insightful understanding into the proactivity of human agency (Seligman, 1999; Seligman and Csíkszentmihályi, 2000). Drawing from this emphasis, we have developed the theory of optimization, which attempts to explain the achievement of optimal functioning in life (e.g., optimal cognitive functioning: academic performance). By the same token, in the course of our research development into the theory of optimization, we have also delved into a comparable theoretical orientation, namely: the multifaceted nature of mindfulness, consisting of three interrelated components - the psychological component of mindfulness, the philosophical component of mindfulness, and the spiritual component of mindfulness. This conceptualization of mindfulness is rather unique for its incorporation of both Western and Eastern knowledge, philosophical viewpoints, and epistemologies into one holistic framework. The main premise of this conceptual analysis article is to advance the study of positive psychology by specifically introducing our recently developed model of mindfulness, in this case, the multifaceted structure of mindfulness with its three distinct components. Importantly, we make attempts to highlight the significance of this multifaceted model by situating it within the theory of optimization for academic learning. Using philosophical psychology and personal-based teaching and research reasoning, we provide a valid rationale as to how aspects of our proposed model of mindfulness (e.g., reaching a state of enlightenment) could act to facilitate and optimize a person's state of functioning (e.g., cognitive functioning). Moreover, we posit that our rationale regarding mindfulness as a potential "optimizing agent" for the purpose of optimal functioning could, indeed, emphasize and reflect the salient nature of positive psychology. In other words, we contend that an explanatory account of mindfulness from the perspectives of Confucianism and Buddhism could, in this analysis, coincide with and support the meaningful understanding and appreciation for the study of positive psychology in educational and non-educational contexts. We conclude the article by exploring the complex issue of methodology - that is, for example, how would a researcher measure, assess, and/or empirically validate the multifaceted nature of mindfulness?

[Newest Reaserch Advance on Targeting Glycolysis Pathways in Acute Myeloid Leukemia --Review].

Metabolic reprogramming plays an important role in the occurrence, drug resistance and recurrence of acute myeloid leukemia (AML). Warburg effect is considered to be the most fundamental metabolic alternation in the process of tumor transformation. The enhancement of glycolytic activation was being found in most malignancies including AML, which promotes anabolism and closely relates with the occurrence and development of tumors. Each metabolic process of glycolysis pathway is catalyzed by specific enzymes. The dependence of AML cells on glycolysis makes targeting inhibition of related catalytic enzymes and interference with glycolysis process a promising therapeutic option. In this review, the recent advances in glycolysis pathway-related targeted therapy in AML are summarized.

Predicting and enhancing students' positive emotions: An empirical study from a Taiwanese sociocultural context.

Predicting and enhancing positive emotions, reflecting the tenets of positive psychology, is of considerable importance for educators. Positive emotions may consist of a person's indication of happiness, joy, and love. In school and university settings, as research has shown, positive emotions play a pivotal role in helping students adjust, make friends, and engage in proactive social relationships with others. It is imperative for us to consider the design and development of educational programs that could assist in the facilitation of positive emotions. The present study involved examination of an inquiry into the prediction of positive emotion of university students, via means of data drawn from Taiwan. The main question for consideration is to determine the extent to which both social (e.g., the social milieu) and personal (e.g., personal resolve) influences could predict positive emotions. Structural equation modelling yielded support for our proposed a priori model: (a) the direct predictive effects of the social milieu, personal resolve, relating to others, and academic liking experience, and (b) the potential mediating roles of relating to others, and academic liking experience. Overall, the concept of emotions plays a central role and is accounted for by different types of personal and social contextual influences.

Achieving optimal best practice: An inquiry into its nature and characteristics.

The study of optimal best practice within the context of academia has produced both empirical and theoretical contributions. Optimal best practice, also coined as optimal functioning, is concerned with a person's personal best-that is, "what is the best that I can do for this academic subject?" Research in the social sciences has, to date, explored different types of optimal best-physical, cognitive, emotional, and social. What is of considerable interest, as a related matter, is a question of how a person reaches and experiences a level of optimal best practice. Recent research development, for example, has explored various conceptualizations of optimal best practice-for example, one distinctive theoretical model, the Framework of Achievement Bests [1, 2], makes a concerted effort to explore the underlying process of optimization-that is, in this case, how an optimal level of best practice is achieved. The present study, as detailed below, investigates via means of non-experimental data a theoretical model pertaining to the achievement of optimal best practice. This examination, we postulate, would enable us to add clarity and provide additional theoretical insights the operational nature of the process of optimization. The operational nature of optimization, as described in our recent research [1, 3], emphasizes three major tenets: (i) the main sources of a person's optimal best practice, (ii) the potential 'optimizing' influences of three comparable agencies on the achievement of optimal best practice (i.e., personal resolve, social relationship, and personal self-efficacy), and (iii) the impact of optimal best practice on future adaptive outcomes (i.e., academic striving and personal well-being). We explored this topic via means of the use of a non-experimental, correlational design with participants drawn from Taiwanese university students (N = 1010). Structural equation modelling (SEM) produced evidence, which empirically supported existing research [1, 3] and substantiated our knowledge of the concept of optimal best practice. Evidence established from the present study has also assisted us to identify one pervasive issue, which we call for further research development-namely, to consider, design, and develop an appropriate methodological approach that would enable researchers to accurately measure and assess the process of optimization. Finally, in terms of teaching and learning, we acknowledge that our research investigation has provided some insights into potential educational practices for implementation.

Understanding levels of best practice: An empirical validation.

Recent research has explored the nature of the theoretical concept of optimal best practice, which emphasizes the importance of personal resolve, inner strength, and the maximization of a person's development, whether it is mental, cognitive, social, or physical. In the context of academia, the study of optimal functioning places emphasis on a student's effort expenditure, positive outlook, and determination to strive for educational success and enriched subjective well-being. One major inquiry closely associated with optimal functioning is the process of optimization. Optimization, in brief, delves into the enactment of different psychological variables that could improve a person's internal state of functioning (e.g., cognitive functioning). From a social sciences point of view, very little empirical evidence exists to affirm and explain a person's achievement of optimal best practice. Over the past five years, we have made extensive progress in the area of optimal best practice by developing different quantitative measures to assess and evaluate the importance of this theoretical concept. The present study, which we collaborated with colleagues in Taiwan, involved the use of structural equation modeling (SEM) to analyze a cohort of Taiwanese university students' (N = 1010) responses to a series of Likert-scale measures that focused on three major entities: (i) the importance of optimal best practice, (ii) three major psychological variables (i.e., effective functioning, personal resolve, and emotional functioning) that could optimize student' optimal best levels in academic learning, and (iii) three comparable educational outcomes (i.e., motivation towards academic learning, interest in academic learning, and academic liking experience) that could positively associate with optimal best practice and the three mentioned psychological variables. Findings that we obtained, overall, fully supported our initial a priori model. This evidence, in its totality, has made substantive practical, theoretical, and methodological contributions. Foremost, from our point of view, is clarity into the psychological process of optimal best practice in the context of schooling. For example, in relation to subjective well-being experiences, how can educators optimize students' positive emotions? More importantly, aside from practical relevance, our affirmed research inquiry has produced insightful information for further advancement. One distinction, in this case, entails consideration of a more complex methodological design that could measure, assess, and evaluate the impact of optimization.

[Clinical analysis of trigemino-cardiac reflex during surgery for patients with skull base tumors].

OBJECTIVE: To investigate the clinical features, risky factors and outcome of the trigemino-cardiac reflex (TCR) during surgery for skull base tumors. METHODS: Two hundred and sixty-two neurosurgical patients with skull base tumors underwent general anesthesia and open surgery from October 2009 to December 2011 in department of neurosurgery of Beijing Tiantan Hospital. The occurrence of TCR and the type of tumor, the surgical approach as well as the postoperative complication relative to TCR was evaluated retrospectively. RESULTS: Seventeen patients occurred TCR events intraoperatively (6.5%). There were 8 men and 9 women with an average age of 40.5 years. Eleven of them (64.7%) underwent schwannoma surgery. Regarding with the surgical procedure, the suboccipital retrosigmoidal approach and the middle fossa transtentorial approach were most commonly associated with TCR in this series (88.2%). The heart rate and blood pressure returned to the patient's normal baseline level after cessation of the surgical manipulation. There was no TCR-relative complication in cardiovascular system. The postoperative course is uneventful in all 17 patients. CONCLUSIONS: TCR may occur during surgery for skull base tumor, especially when performing schwannoma surgery and suboccipital retrosigmoidal or middle fossa transtentorial approach. Accurate recognition and management of TCR during skull base surgery often carry on favorable outcome.

M9, a novel region of amino-Nogo-A, attenuates cerebral ischemic injury by inhibiting NADPH oxidase-derived superoxide production in mice.

AIMS: In acute stroke, neurological damage is due to oxidative stress and neuronal apoptotic death. This study investigated whether Nogo-A 290-562 residues region (M9), fused to the transduction domain of the HIV trans-activator (TAT) protein, is neuroprotective against cerebral ischemia and the mechanisms. METHODS: Transient focal cerebral ischemia was induced by middle cerebral artery occlusion in male C57BL/6J mice. TAT-M9, its mutation or vehicle was applied via intraperitoneal injection at the onset of reperfusion. The neurobehavioral scores, infarction volumes, neuronal apoptosis, and the ratio of Bax/Bcl-2 were evaluated. Malondialdehyde (MDA), reactive oxygen species (ROS) levels, and NADPH oxidase activation were measured in the presence or absence of the NADPH oxidase inhibitor apocynin or activator tetrabromocinnamic acid (TBCA). RESULTS: Immunofluorescence results confirmed that TAT-M9 was transduced into brain parenchyma, and it significantly improved neurological behavior, reduced infarct volumes, protected neuronal cells from apoptosis, inhibited activation of NADPH oxidase, and decreased MDA and ROS contents. Furthermore, apocynin imitated the beneficial effects of TAT-M9, while TBCA abolished them. CONCLUSIONS: Our results demonstrate that TAT-M9 administration attenuates cerebral ischemia by inhibiting NADPH oxidase-mediated oxidative damage and neuronal apoptosis in mice. TAT-M9 may be a potential treatment for cerebrovascular disease.

[Effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels in a rabbit model of lung ischemia-reperfusion injury].

OBJECTIVE: To study the effects of limb ischemia preconditioning on pulmonary free radicals and cytokine levels during lung ischemia-reperfusion injury in rabbits. METHODS: Eighteen healthy rabbits were randomly divided into three groups: control group (group C, n = 6), ischemia/reperfusion group (group I/R, n = 6), limb ischemia preconditioning group (group L, n = 6). At the end of experiments, the wet to dry-weight ratio (W/D), activities of superoxide dismutase (SOD) and myeloperoxidase (MPO), levels of malondialdehyde (MDA) and the contents of cytokines (TNF-α, IL-6, IL-8 and IL-10) were determined in lung tissues. Protein levels of bronchoalveolar lavage fluid and serum were measured to calculate the lung permeability index. Pathologic changes of lung tissues were also observed. RESULTS: Compared to the group I/R, the lung tissue W/D ratio, MPO activity, lung permeability index, MDA and the cytokines (TNF-α, IL-6 and IL-8) levels were significantly decreased in group L (P < 0.05), while the SOD activity (P < 0.05) and IL-10 contents were significantly increased (P < 0.01). There was no statistical difference in the changes of the above parameters between group L and group C (P > 0.05). The morphologic damages were significantly reduced in group L than that in group I/R. CONCLUSION: Limb ischemia preconditioning has protective effect against lung ischemia-reperfusion injury, which may at least in part through inhibiting the release of oxygen-derived free radicals and pro-inflammatory cytokines (TNF-α, IL-6, IL-8) and increasing the production of anti-inflammatory cytokine IL-10.

Soybean GmPHD-type transcription regulators improve stress tolerance in transgenic Arabidopsis plants.

BACKGROUND: Soybean [Glycine max (L.) Merr.] is one of the most important crops for oil and protein resource. Improvement of stress tolerance will be beneficial for soybean seed production. PRINCIPAL FINDINGS: Six GmPHD genes encoding Alfin1-type PHD finger protein were identified and their expressions differentially responded to drought, salt, cold and ABA treatments. The six GmPHDs were nuclear proteins and showed ability to bind the cis-element "GTGGAG". The N-terminal domain of GmPHD played a major role in DNA binding. Using a protoplast assay system, we find that GmPHD1 to GmPHD5 had transcriptional suppression activity whereas GmPHD6 did not have. In yeast assay, the GmPHD6 can form homodimer and heterodimer with the other GmPHDs except GmPHD2. The N-terminal plus the variable regions but not the PHD-finger is required for the dimerization. Transgenic Arabidopsis plants overexpressing the GmPHD2 showed salt tolerance when compared with the wild type plants. This tolerance was likely achieved by diminishing the oxidative stress through regulation of downstream genes. SIGNIFICANCE: These results provide important clues for soybean stress tolerance through manipulation of PHD-type transcription regulator.

An R2R3-type transcription factor gene AtMYB59 regulates root growth and cell cycle progression in Arabidopsis.

MYB proteins play important roles in eukaryotic organisms. In plants, the R1R2R3-type MYB proteins function in cell cycle control. However, whether the R2R3-type MYB protein is also involved in the cell division process remains unknown. Here, we report that an R2R3-type transcription factor gene, AtMYB59, is involved in the regulation of cell cycle progression and root growth. The AtMYB59 protein is localized in the nuclei of onion epidermal cells and has transactivation activity. Expression of AtMYB59 in yeast cells suppresses cell proliferation, and the transformants have more nuclei and higher aneuploid DNA content with longer cells. Mutation in the conserved domain of AtMYB59 abolishes its effects on yeast cell growth. In synchronized Arabidopsis cell suspensions, the AtMYB59 gene is specifically expressed in the S phase during cell cycle progression. Expression and promoter-GUS analysis reveals that the AtMYB59 gene is abundantly expressed in roots. Transgenic plants overexpressing AtMYB59 have shorter roots compared with wild-type plants (Arabidopsis accession Col-0), and around half of the mitotic cells in root tips are at metaphase. Conversely, the null mutant myb59-1 has longer roots and fewer mitotic cells at metaphase than Col, suggesting that AtMYB59 may inhibit root growth by extending the metaphase of mitotic cells. AtMYB59 regulates many downstream genes, including the CYCB1;1 gene, probably through binding to MYB-responsive elements. These results support a role for AtMYB59 in cell cycle regulation and plant root growth.

[Effect of fluid resuscitation for hemorrhagic shock on oxygenation of subjects in early period of first visit of high altitude area, an experimentation on dogs].

OBJECTIVE: To investigate the effect of fluid resuscitation on oxygenation of subjects with hemorrhagic shock in early period of first visit to area of high altitude, an experiment in dogs was performed. METHODS: A model of serious hemorrhagic shock was reproduced by exsanguination resulting in a lowering of mean arterial pressure to (35+/-5) mm Hg (1 mm Hg=0.133 kPa) maintain for 1 hour. Thirteen mongrel dogs were carried to an area of 3,780 metres above sea level from an area of 1,510 metres, and they were randomly divided into three groups, namely lactated Ringer's (LR) group, 6% hydroxyethyl starch (HES) group, and control group. The dogs in LR group were infused intravenously LR in 1.5 times the volume of blood loss; those in 6% HES group were given HES in equal volume. No fluid infusion was given in the control group. After 1 hour of resuscitation, LR was intravenously given at 5 ml.kg(-1).h(-1) in all groups as maintenance dose. The changes in oxygenation were observed. RESULTS: All animals in control group were dead after 2 hours. One hour after establishment of shock, the oxygen consumption (VO(2)), oxygen delivery (DO(2)), oxygen extraction ratio (O(2)ER), arterial oxygen saturation (SaO(2)) in two resuscitation groups were significantly lower than those before shock, but venous oxygen saturation (Sv(2)) and alveolar-arterial oxygen difference (A-aDO(2)) were significantly higher (all P<0.05). In LR group, the oxygenation parameters including VO(2), DO(2), O(2)ER, SaO(2) after 2 hours of resuscitation were significantly higher than those 1 hour after shock, while A-aDO(2) was significantly lower (all P<0.05); and in HES group, VO(2), DO(2), O(2)ER were significantly higher than those 1 hour after shock, while SvO(2) was significantly lower (all P<0.05). CONCLUSION: All dogs with serious hemorrhagic shock would die of hemorrhagic shock in the early period of entering a high altitude area if fluid resuscitation is denied. Two hours after infusion of LR in 1.5 times of quantity of blood loss, oxygenation can be restored to expected normal values. Infusion of 6% HES with an equal volume of blood loss, oxygenation dose not reach expected level 2 hours after resuscitation.

Soybean GmMYB76, GmMYB92, and GmMYB177 genes confer stress tolerance in transgenic Arabidopsis plants.

MYB-type transcription factors contain the conserved MYB DNA-binding domain of approximately 50 amino acids and are involved in the regulation of many aspects of plant growth, development, metabolism and stress responses. From soybean plants, we identified 156 GmMYB genes using our previously obtained 206 MYB unigenes, and 48 were found to have full-length open-reading frames. Expressions of all these identified genes were examined, and we found that expressions of 43 genes were changed upon treatment with ABA, salt, drought and/or cold stress. Three GmMYB genes, GmMYB76, GmMYB92 and GmMYB177, were chosen for further analysis. Using the yeast assay system, GmMYB76 and GmMYB92 were found to have transactivation activity and can form homodimers. GmMYB177 did not appear to have transactivation activity but can form heterodimers with GmMYB76. Yeast one-hybrid assay revealed that all the three GmMYBs could bind to cis-elements TAT AAC GGT TTT TT and CCG GAA AAA AGG AT, but with different affinity, and GmMYB92 could also bind to TCT CAC CTA CC. The transgenic Arabidopsis plants overexpressing GmMYB76 or GmMYB177 showed better performance than the GmMYB92-transgenic plants in salt and freezing tolerance. However, these transgenic plants exhibited reduced sensitivity to ABA treatment at germination stage in comparison with the wild-type plants. The three GmMYB genes differentially affected a subset of stress-responsive genes in addition to their regulation of a common subset of stress-responsive genes. These results indicate that the three GmMYB genes may play differential roles in stress tolerance, possibly through regulation of stress-responsive genes.

Soybean WRKY-type transcription factor genes, GmWRKY13, GmWRKY21, and GmWRKY54, confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants.

WRKY-type transcription factors have multiple roles in the plant defence response and developmental processes. Their roles in the abiotic stress response remain obscure. In this study, 64 GmWRKY genes from soybean were identified, and were found to be differentially expressed under abiotic stresses. Nine GmWRKY proteins were tested for their transcription activation in the yeast assay system, and five showed such ability. In a DNA-binding assay, three proteins (GmWRKY13, GmWRKY27 and GmWRKY54) with a conserved WRKYGQK sequence in their DNA-binding domain could bind to the W-box (TTGAC). However, GmWRKY6 and GmWRKY21, with an altered sequence WRKYGKK, lost the ability to bind to the W-box. The function of three stress-induced genes, GmWRKY13, GmWRKY21 and GmWRKY54, was further investigated using a transgenic approach. GmWRKY21-transgenic Arabidopsis plants were tolerant to cold stress, whereas GmWRKY54 conferred salt and drought tolerance, possibly through the regulation of DREB2A and STZ/Zat10. Transgenic plants over-expressing GmWRKY13 showed increased sensitivity to salt and mannitol stress, but decreased sensitivity to abscisic acid, when compared with wild-type plants. In addition, GmWRKY13-transgenic plants showed an increase in lateral roots. These results indicate that the three GmWRKY genes play differential roles in abiotic stress tolerance, and that GmWRKY13 may function in both lateral root development and the abiotic stress response.

The soybean Dof-type transcription factor genes, GmDof4 and GmDof11, enhance lipid content in the seeds of transgenic Arabidopsis plants.

Soybean is one of the most important leguminous seed crops among the oil crops. Although the pathways for lipid biosynthesis have been identified, the factors that regulate the biosynthetic pathways at the transcriptional level are largely unknown. Here, we report our findings on the involvement of soybean Dof-type transcription factor genes in the regulation of the lipid content in soybean seeds. We identified 28 Dof-type transcription factor genes in soybean plants, and these genes displayed diverse patterns of expression in various organs. Seven flower/pod-specific genes and one constitutively expressed gene were further investigated. The proteins encoded by these seven genes were localized in the nucleus, and exhibited different abilities for transcriptional activation and DNA binding. Two genes, GmDof4 and GmDof11, were found to increase the content of total fatty acids and lipids in GmDof4 and GmDof11 transgenic Arabidopsis seeds. We also found that the 1000-seed weight was increased in the GmDof4 and GmDof11 transgenic plants. Using microarray and DNA binding analysis, we found that the two Dof-like proteins, GmDof4 and GmDof11, activated the acetyl CoA carboxylase gene and long-chain-acyl CoA synthetase gene, respectively, by direct binding to the cis-DNA elements in their promoter regions. In addition, both proteins downregulated the storage protein gene, CRA1, through direct binding. These results suggest that the two GmDof genes may augment the lipid content of soybean seeds by upregulating genes that are associated with the biosynthesis of fatty acids.

Cloning and comparative analysis of the gene encoding diacylglycerol acyltransferase from wild type and cultivated soybean.

Diacylglycerol acyltransferase (DGAT), as an important enzyme in triacylglycerol synthesis, catalyzes the final acylation of the Kennedy pathway. In the present study, the GmDGAT gene was cloned from Glycine max by using AtDGAT as a query to search against the soybean EST database and the rapid amplification of cDNA ends (RACE) method. Allelic genes were also isolated from 13 soybean accessions and the divergence of the deduced amino acid sequences were compared. The comparison reveals that although GmDGAT is a highly conserved protein, several differences of insertion/deletion were identified in the N-terminal region of the GmDGATs from various soybean accessions. In the C-terminal regions, a single amino acid mutation specific to both G. max and G. soja was also found. The GmDGAT genomic sequences were further cloned and the number and size of exons in the DGAT genomic sequence were very similar among different plant species, whereas the introns were more diverged. These results may have significance in elucidating the genetic diversity of the GmDGAT among the soybean subgenus.