Exogenous citric acid enhances drought tolerance in tobacco (Nicotiana tabacum).

Organic acids play a pivotal role in improving plant response to long-term drought stress. External application of organic acids has been reported to improve drought resistance in several species. However, whether organic acids have similar effects in tobacco remains unknown. A screening study of the protective function of organic acids in tobacco and understanding the underlying molecular mechanism would be useful in developing a strategy for drought tolerance. Several physiological and molecular adaptations to drought including abscisic acid, stomatal closure, reactive oxygen species homeostasis, amino acid accumulation, and drought-responsive gene expression were observed by exogenous citric acid in tobacco plants. Exogenous application of 50 mm citric acid to tobacco plants resulted in higher chlorophyll content, net photosynthesis, relative water content, abscisic acid content and lower stomatal conductance, transpiration and water loss under drought conditions. Moreover, reactive oxygen species homeostasis was better maintained through increasing activity of antioxidant enzymes and decreasing hydrogen peroxide content after citric acid pretreatment under drought. Amino acids involved in the TCA cycle accumulated after external application of citric acid under drought stress. Furthermore, several drought stress-responsive genes also dramatically changed after application of citric acid. These data support the idea that external application of citric acid enhances drought resistance by affecting physiological and molecular regulation in tobacco. This study provides clear insights into mechanistic details of regulation of amino acid and stress-responsive gene expression by citric acid in tobacco in response to drought, which is promising for minimizing growth inhibition in agricultural fields.

Extending chemotherapy with capecitabine following CAPOX chemotherapy improves survival of Stage 3 gastric carcinoma after radical surgery: a 5-year analysis.

OBJECTIVE: To assess the effectiveness and safety of treatment consisting of extending chemotherapy (ECT) with capecitabine following capecitabine plus oxaliplatin (CAPOX) chemotherapy for stage 3 gastric carcinoma (GC) after D2 gastrectomy. PATIENTS AND METHODS: In this retrospective study, we included 214 patients with stage 3 GC who underwent D2 gastrectomy between January 2012 and April 2014. The CAPOX regimen chemotherapy was administrated to all of the patients as adjuvant therapy. The CAPOX regimen consisted of capecitabine (1000 mg/m2, in 2 divided doses for 14 d) and oxaliplatin (130 mg/m2 given on Day 1), repeated every 21 d for 8 cycles. Following CAPOX chemotherapy, 102 of these patients received extending chemotherapy (the ECT group) with capecitabine, whereas 112 patients (the control group) received no ECT. The ECT consisted of capecitabine (1000 mg/m2, in 2 divided doses for 14 d), repeated every 21 d for 8 cycles at most. The chemotherapy was discontinued if unacceptable toxicity or disease progression occurred or upon the request of the patient. All cases were followed up, and overall survival (OS), recurrence-free survival (RFS), and toxicities were compared. RESULTS: The ECT group exhibited a distinctly higher 5-year OS (p=0.0468) and RFS (p=0.0483) than those of the control group. The incidence of hand-foot syndrome was markedly greater in the ECT group (p=0.0043). No toxicity-related death occurred. CONCLUSIONS: Extending chemotherapy with capecitabine following the CAPOX regimen chemotherapy provides significant survival benefit for stage 3 GC after D2 gastrectomy.

Effect of Sevelamer on Calciprotein Particles in Hemodialysis Patients: The Sevelamer Versus Calcium to Reduce Fetuin-A-Containing Calciprotein Particles in Dialysis (SCaRF) Randomized Controlled Trial.

INTRODUCTION: Calciprotein particles (CPPs) are potentially modifiable mediators of phosphate toxicity in patients with kidney disease. We compared the effects of calcium carbonate (CC) and the non-calcium-based phosphate binder sevelamer on CPP levels in patients undergoing hemodialysis (HD). We hypothesized that treatment with sevelamer would achieve greater reductions in amorphous calcium phosphate-containing CPP (CPP-1) and hydroxyapatite-containing CPP (CPP-2) owing to reduced calcium loading and anti-inflammatory pleiotropic effects. METHODS: We conducted an open-label, randomized controlled trial (RCT) in which 31 stable prevalent HD patients were allocated to receive either sevelamer hydrochloride (SH), sevelamer carbonate (SC), or CC for 24 weeks. Dual primary endpoints were the between groups differences in serum CPP-1 and CPP-2 levels at 24 weeks in SH + SC-treated versus CC-treated patients. Effects on aortic pulse wave velocity (aPWV), inflammatory cytokines (interleukin-6 and -8), and effects across individual treatment arms were also assessed. RESULTS: Serum CPP-1, but not CPP-2, levels were lower in those randomly assigned to the sevelamer (SH + SC) group compared with the CC group at 24 weeks (-70%, 95% confidence interval [CI] -90% to -15%, P = 0.02). In subgroup analysis, this effect was confined to those receiving SC (-83.4%, 95% CI -95.7% to -36.8%, P = 0.01). aPWV and interleukin-8 levels were also lower in those who received sevelamer compared with CC at 24 weeks (-2.0 m/s, 95% CI -2.9 to -1.1; -57%, 95% CI -73% to -30%, respectively, both P = 0.01). Conventional markers of mineral metabolism remained stable across all treatment groups. DISCUSSION: Compared with treatment with CC, use of sevelamer for 24 weeks was associated with lower serum CPP-1 levels and a reduction in aPWV and systemic inflammation.

MiR-201-5p alleviates lipopolysaccharide-induced renal cell dysfunction by targeting NOTCH3.

OBJECTIVE: Lipopolysaccharide (LPS)-induced inflammation and dysfunction in the kidney may be the major risk factors for subsequent acute kidney injury (AKI). Previous studies have reported that up-regulation of notch receptor 3 (NOTCH3) expression is accompanied with renal epithelium and podocyte damage. Herein, we aimed to investigate whether NOTCH3 was involved in lipopolysaccharide (LPS)-induced AKI and renal cell dysfunction. MATERIALS AND METHODS: Septic mice were established using LPS (20 mg/kg) intraperitoneally. mRNA and protein expression in the kidney and renal cell was performed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blotting, respectively. Cell counting kit-8 (CCK8) and flow cytometry were used to measure cell viability and apoptosis, respectively. Bioinformatics algorithm and Luciferase reporter gene assay were performed to validate whether NOTCH3 was a direct target of miR-201-5p. RESULTS: Up-regulation of NOTCH3 and down-regulation of miR-201-5p were observed in the kidney of LPS-induced septic mice. LPS-stimulated TCMK-1 and MPC5 cells led to an increase in NOTCH3 and a decrease in miR-201-5p expression levels. Bioinformatics algorithm and experimental measurements validated that NOTCH3 was a direct target of miR-201-5p. Overexpression of miR-201-5p protected against LPS-induced renal cell growth inhibition, apoptosis and inflammatory response via the suppression of toll-like receptor 4 (TLR4)/NOTCH3 signaling pathway. CONCLUSIONS: The novel role of miR-201-5p via the inhibition of LPS-activated TLR4/NOTCH3 might provide a potential therapeutic strategy for the treatment of LPS-induced AKI.

[The expression and clinical progression of androgen, estrogen and prolactin receptor in laryngeal squamous cell carcinoma].

Objective:The aim of this study is to analyze the different expression and function of androgen receptors (AR) and estrogen receptor (ER-α) in laryngeal squamous cell carcinoma (LSCC). In combination with the expression of prolactin receptor (PRLR), we analyzed the prognostic impact of three receptors on the laryngeal carcinoma. Method:In this study, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PC) analysis were performed on the tumor tissues and adjacent normal tissues in 112 LSCC patients (95 males and 17 females). We found that hormone receptor expression is closely related to the clinical tumor lesions and overall survival data. Result:The expression of AR, ER-α and PRLR in tumor tissues were much higher than those in adjacent normal tissues (P>0.05) at both protein and mRNA levels. The higher PRLR level indicate poor survival in LSCC patients (P=0.02), while higher ER-α expression could influence the survival with considerable trend toward significance (P=0.06). Furthermore, the higher expression of ER-α in tumours were corresponding with PRLR cytoplasmic higher level expression (r=0.802, P=0.04). This mutual promoted effect between ER-α and PRLR possibly suggests potential mechanisms among those sex related hormone receptors in laryngeal cancer. Conclusion:It has become increasingly credible that the sex related hormone receptors play an important role in the development of LSCC.

Autologous hematopoietic cell transplantation for multiple myeloma patients with renal insufficiency: a center for international blood and marrow transplant research analysis.

Autologous hematopoietic cell transplantation (AHCT) in multiple myeloma (MM) patients with renal insufficiency (RI) is controversial. Patients who underwent AHCT for MM between 2008 and 2013 were identified (N=1492) and grouped as normal/mild (⩾60 mL/min), N=1240, moderate (30-59), N=185 and severe RI (<30), N=67 based on Modification of Diet in Renal Disease. Multivariate analyses of non-relapse mortality (NRM), relapse, PFS and overall survival (OS) were performed. Of the 67 patients with severe RI, 35 were on dialysis prior to AHCT. Patients received melphalan 200 mg/m2 (Mel 200) in 92% (normal/mild), 75% (moderate) and 33% (severe) RI; remainder received 140 mg/m2 (Mel 140). Thirty four of 35 patients with severe RI achieved post-AHCT dialysis independence. The 5-year PFS for normal, moderate and severe RI was 35 (95% CI, 31-38)%, 40 (31-49)% and 27 (15-40)%, respectively, (P=0.42); 5-year OS for normal, moderate and severe RI was 68 (65-71)%, 68 (60-76)% and 60 (46-74)%, respectively, (P=0.69). With moderate RI, 5-year PFS for high-dose melphalan 140 mg/m2 was 18 (6-35)% and for Mel 200 was 46 (36-57)% (P=0.009). With severe RI, 5-year PFS Mel 140 was 25 (11-41) % and for Mel 200 was 32 (11-58)% (P=0.37). We conclude that AHCT is safe and effective in patients with MM with RI.

Comparison of effectiveness of two commonly used two-handed mask ventilation techniques on unconscious apnoeic obese adults.

BACKGROUND.: Mask ventilation and tracheal intubation are basic techniques for airway management and mutually inclusive rescue measures to restore ventilation. The aim of this study was to compare the effectiveness of mask ventilation between two commonly used techniques of two-handed mask ventilation in obese unconscious apnoeic adults. METHODS.: Eighty-one obese adults received mask ventilation after induction using C-E clamp and modified V-E clamp techniques in a randomized crossover manner. Mechanical ventilation was provided using a pressure-control mode, at a rate of 10 bpm, with an inspiratory-to-expiratory time ratio of 1:2 and a pre-set plateau airway pressure of 20 cm H 2 O. The primary outcome was expired tidal volume. RESULTS.: The BMI for the subjects was 37 ( sd 4.9) kg m -2 . The failure rates for mask ventilation (tidal volume≤anatomical dead space) were 44% for the C-E technique and 0% for the V-E technique ( P <0.001). Tidal volume was significantly lower for the C-E than the V-E technique [371 ( sd 345) vs 720 (244) ml, P <0.001]. The peak airway pressures were 21 ( sd 1.5) cm H 2 O for the C-E technique and 21 (1.3) cm H 2 O for the V-E technique. CONCLUSIONS.: Mask ventilation using the modified V-E technique is more effective than with the C-E technique in unconscious obese apnoeic adults. Subjects who fail ventilation with the C-E technique can be ventilated effectively with the V-E technique. CLINICAL TRIAL REGISTRATION.: NCT02580526.

[Preliminary findings for metabolite profiles of papillary thyroid carcinoma and laryngeal squamous cell carcinoma].

Objective:To identify distinct metabolite profiles of papillary thyroid cancer (PTC) and laryngeal squamous cell carcinoma (LSCC).Method:Tumor and adjacent non-tumor specimens were collected from 57 PTC and 33 LSCC patients. Distinct metabolite profiles of tissues were examined using a combination of gas chromatography-time-of-flight mass spectrometry and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. The data were analyzed with multivariate statistical analysis to compare the distinct metabolite profiles and related pathways of these three tumors.Result:A panel of 46 and 41 differentially expressed metabolites were identified in tumor and adjacent non-tumor tissues of PTC and LSCC, respectively. Increased glycolysis, amino acids metabolism, one carbon metabolism and tryptophan metabolism were found in these two types of tumor tissues compared to adjacent non tumor tissues. Purine and pyrimidine metabolism was significantly elevated in PTC and LSCC tumor tissues, while taurine and hypotaurine were only higher in PTC tumor tissues. The fatty acid metabolism was detected at lower level in both PTC and LSCC tumor tissue.Conclusion:PTC and LSCC tumor tissues not only have common metabolic signatures characterized by increased glycolysis, amino acids metabolism, one carbon metabolism and tryptophan metabolism, but also have distinct metabolic signatures. It is helpful to understand the nature of these two tumors.

A Novel Cooling Device for Targeted Brain Temperature Control and Therapeutic Hypothermia: Feasibility Study in an Animal Model.

BACKGROUND: Therapeutic hypothermia (i.e., temperature management) is an effective option for improving survival and neurological outcome after cardiac arrest and is potentially useful for the care of the critically ill neurological patient. We analyzed the feasibility of a device to control the temperature of the brain by controlling the temperature of the blood flowing through the neck. METHODS: A lumped parameter dynamic model, with one-dimensional heat transfer, was used to predict cooling effects and to test experimental hypotheses. The cooling system consisted of a flexible collar and was tested on 4 adult sheep, in which brain and body temperatures were invasively monitored for the duration of the experiment. RESULTS: Model-based simulations predicted a lowering of the temperature of the brain and the body following the onset of cooling, with a rate of 0.4 °C/h for the brain and 0.2 °C/h for the body. The experimental findings showed comparable cooling rates in the two body compartments, with temperature reductions of 0.6 (0.2) °C/h for the brain and 0.6 (0.2) °C/h for the body. For a 70 kg adult human subject, we predict a temperature reduction of 0.64 °C/h for the brain and 0.43 °C/h for the body. CONCLUSIONS: This work demonstrates the feasibility of using a non-invasive method to induce brain hypothermia using a portable collar. This device demonstrated an optimal safety profile and represents a potentially useful method for the administration of mild hypothermia and temperature control (i.e., treatment of hyperpyrexia) in cardiac arrest and critically ill neurologic patients.

[The characteristic of metabolics of female papillary thyroid carcinoma and its relationship with primary tumor size and cervical lymph node metastases].

OBJECTIVE: To analyze the metabolic profiles of the female papillary thyroid carcinoma (PTC) and the relationship between the metabolic profiles and primary tumor size and cervical lymph node metastasis using a metabolomics approach. METHODS: Forty-three cases of female PTC were enrolled in this study. Gas chromatography-time-of-flight mass spectrometry and ultra performance liquid chromatography-time of flight-mass spectrometry were employed for analyzing metabolic profiles of tumor tissues and adjacent normal tissues in the female PTC. Cases were divided into Group T1 (tumor size≤2.0 cm) and Group T2 (tumor size>2.0 cm) according to the tumor size, and divided into Group N- (with negative cervical lymph node) and Group N+ (with positive cervical lymph node) according to the cervical lymph node conditions. We compared the metabolomic profiles between these groups. RESULTS: A panel of 46 differentially expressed metabolites was identified in the PTC specimens, compared with normal tissues. Increased metabolism of amino acid, purine and pyrimidine, tryptophan acid, one carbon, glycolysis, taurine and hypotaurine, and fatty acid were found in PTC tumors tissues. Amino acids, purine and pyridine, tryptophan, and carbon metabolism increased significantly in the tumor tissues of Group T2 compared with Group T1, while glycolysis, amino acid, purine and pyridine, tryptophan, and carbon metabolism increased significantly in the specimens of Group N+ . CONCLUSION: Distinct metabolic profiles were identified in the female PTC tissues, which were related to the primary tumor size and cervical lymph node metastases.

Expression of putative stem marker nestin and CD133 in advanced serous ovarian cancer.

It is hypothesized that "cancer stem cells" are responsible for the resistance to chemotherapy of cancer cells in ovarian cancers. The objective of the studies was to explore if the stem cell biomarkers could be used to predict the tumor chemotherapy-resistance in serous ovarian cancer patients. Expression of two putative stem cell markers CD133 and nestin, and vascular epithelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) were detected in 123 cases of advanced serous ovarian cancer specimens by immunohistochemistry. To estimate intra-tumoral microvessel density (MVD), CD34 immunostaining was also performed. CD133 and nestin were defined to be positive in 35.0% and 32.5% of the serous ovarian carcinoma tissues, respectively. It was observed that overexpression of nestin but not CD133 was associated with the cisplatin-based chemotherapy resistance and shorter overall survival of the patients, and nestin was found to be an independent prognostic factor. Moreover, positive nestin expression also correlated to increased expression of EGFR and VEGF, and elevated MVD in tumors. The results of this study suggest that serous ovarian cancers with high expression level of nestin represent an aggressive malignant phenotype associated with poor prognosis, and treatment targeted the nestin positive cancer cells might be a promising therapeutic strategy for this subgroups.

Predicting mortality from HIV-associated Pneumocystis pneumonia at illness presentation: an observational cohort study.

BACKGROUND: Although the use of antiretroviral therapy has led to dramatic declines in AIDS-associated mortality, Pneumocystis pneumonia (PCP) remains a leading cause of death in HIV-infected patients. OBJECTIVES: To measure mortality, identify predictors of mortality at time of illness presentation and derive a PCP mortality prediction rule that stratifies patients by risk for mortality. METHODS: An observational cohort study with case note review of all HIV-infected persons with a laboratory diagnosis of PCP at San Francisco General Hospital from 1997 to 2006. RESULTS: 451 patients were diagnosed with PCP on 524 occasions. In-hospital mortality was 10.3%. Multivariate analysis identified five significant predictors of mortality: age (adjusted odds ratio (AOR) per 10-year increase, 1.69; 95% CI 1.08 to 2.65; p = 0.02); recent injection drug use (AOR 2.86; 95% CI 1.28 to 6.42; p = 0.01); total bilirubin >0.6 mg/dl (AOR 2.59; 95% CI 1.19 to 5.62; p = 0.02); serum albumin <3 g/dl (AOR 3.63; 95% CI 1.72-7.66; p = 0.001); and alveolar-arterial oxygen gradient >or=50 mm Hg (AOR 3.02; 95% CI 1.41 to 6.47; p = 0.004). Using these five predictors, a six-point PCP mortality prediction rule was derived that stratifies patients according to increasing risk of mortality: score 0-1, 4%; score 2-3, 12%; score 4-5, 48%. CONCLUSIONS: The PCP mortality prediction rule stratifies patients by mortality risk at the time of illness presentation and should be validated as a clinical tool.

Characterization and application of two novel monoclonal antibodies against 2IgB7-H3: expression analysis of 2IgB7-H3 on dendritic cells and tumor cells.

2IgB7-H3 has recently been identified as a new member of the B7 family. Its expression at the protein level remains largely unknown due to the lack of the specific monoclonal antibody (mAb). To characterize the expression of 2IgB7-H3, we newly generated two mouse antihuman 2IgB7-H3 mAbs (4H7 and 21D4). We found the constitutive expression of 2IgB7-H3 on a series of tumor cell lines. Furthermore, the expression was examined on monocyte-derived dendritic cells (Mo-DCs) and DCs from CD34(+) hematopoietic progenitor cells (HPC) by means of mAb staining. The results showed that 2IgB7-H3 was expressed on Mo-DCs at a high and stable level during differentiation in vitro. With the maturation of DCs from CD34(+) HPCs, the expression of the molecule was upregulated. However, the 2IgB7-H3 was not expressed on fresh isolated T and B lymphocytes, monocytes, or CD34(+) HPCs. These results suggested that 2IgB7-H3 may be a valuable surface antigen for the detection of DCs.

Effects of microinjection of melatonin and its receptor antagonists into anterior hypothalamic area on blood pressure and heart rate in rats.

AIM: To examine the effects of microinjection of melatonin and its receptor antagonists into the anterior hypothalamic area (AHA) on blood pressure (BP) and heart rate (HR) in normotensive and stress-induced hypertensive rats. METHODS: Melatonin and its receptor antagonists were microinjected into the AHA, then BP, mean arterial pressure (MAP), and HR were synchronously recorded. RESULTS: Microinjection of melatonin produced a fall in MAP. Prazosin, an antagonist of melatonin ML2 receptor, could not antagonize the depressive response induced by melatonin. While luzindole, a competitive antagonist of melatonin ML1 receptor, was able to almost completely prevented the depressive response induced by injection of melatonin. CONCLUSION: Melatonin acts as a hypotensive factor and the effects are mainly due to activation of ML1 receptors in rat brain, and the AHA may be one of the important central areas where melatonin can exert modulatory effects on BP and HR.

X-ray structure of azide-bound fully oxidized cytochrome c oxidase from bovine heart at 2.9 A resolution.

Two azide ions were identified, one between the Fe and Cu atoms in the O(2)-reduction site and the other at the transmembrane surface of the enzyme, in the crystal structure of the azide-bound form of bovine heart cytochrome c oxidase at 2.9 A resolution. Two geometries, a mu-1,3 type geometry between the Fe and Cu atoms and a terminal geometry on the Fe atom, are equally possible for an azide ion in the O(2)--reduction site. The other azide molecule was hydrogen bonded to an amide group of an asparagine and a hydroxyl group of tyrosine in a mu-1,1 type geometry. The antisymmetric infrared bands arising from these azide ions, which show essentially identical intensity [Yoshikawa & Caughey (1992), J. Biol. Chem. 267, 9757-9766], strongly suggest terminal binding of the azide to Fe. The electron density of all three imidazole ligands to Cu(B) was clearly seen in the electron-density map of the azide-bound form of bovine heart enzyme, in contrast to the crystal structure of the azide-bound form of the bacterial enzyme [Iwata et al. (1995), Nature (London), 376, 660-669], which lacks one of the three imidazole ligands to Cu(B).

Redox-coupled crystal structural changes in bovine heart cytochrome c oxidase.

Crystal structures of bovine heart cytochrome c oxidase in the fully oxidized, fully reduced, azide-bound, and carbon monoxide-bound states were determined at 2.30, 2.35, 2.9, and 2.8 angstrom resolution, respectively. An aspartate residue apart from the O2 reduction site exchanges its effective accessibility to the matrix aqueous phase for one to the cytosolic phase concomitantly with a significant decrease in the pK of its carboxyl group, on reduction of the metal sites. The movement indicates the aspartate as the proton pumping site. A tyrosine acidified by a covalently linked imidazole nitrogen is a possible proton donor for the O2 reduction by the enzyme.

[Relation between the incidence of ectopic pregnancy and the use of intrauterine devices].

The relationship between the incidence of ectopic pregnancy (EP) and the use of IUD was examined by the method of analytical epidemiological study. 10,843 women of child-bearing age from the west district of Beijing were investigated. The conclusions were: (1) The incidence of EP in IUD users was 0.91/1,000 women year; and the EP incidence in women not using any contraceptives was 2.23/1,000 women year. (2) The incidence of EP in women with IUD inserted within 2 years was significantly higher than that in women using IUD longer than 2 years, that is 5.64/1,000 women year and 0.47/1,000 women year respectively. (3) The probability of suffering from EP for accidental pregnancy in IUD users was 4.84%, which was much higher than that of women not using any contraceptives (0.20%). (4) The risk of EP for IUD users with pelvic inflammatory disease was 6.64 times compared to those without pelvic inflammatory disease. (5) EP was not related to previous cesarean section or induced abortion.