Temporal trends and rural-urban disparities in cerebrovascular risk factors, in-hospital management and outcomes in ischaemic strokes in China from 2005 to 2015: a nationwide serial cross-sectional survey.

BACKGROUND: Stroke is the leading cause of mortality in China, with limited evidence of in-hospital burden obtained from nationwide surveys. We aimed to monitor and track the temporal trends and rural-urban disparities in cerebrovascular risk factors, management and outcomes from 2005 to 2015. METHODS: We used a two-stage random sampling survey to create a nationally representative sample of patients admitted for ischaemic stroke in 2005, 2010 and 2015. We sampled participating hospitals with an economic-geographical region-stratified random-sampling approach first and then obtained patients with a systematic sampling approach. We weighed our survey data to estimate the national-level results and assess changes from 2005 to 2015. RESULTS: We analysed 28 277 ischaemic stroke admissions from 189 participating hospitals. From 2005 to 2015, the estimated national hospital admission rate for ischaemic stroke per 100 000 people increased (from 75.9 to 402.7, Ptrend<0.001), and the prevalence of risk factors, including hypertension, diabetes, dyslipidaemia and current smoking, increased. The composite score of diagnostic tests for stroke aetiology assessment (from 0.22 to 0.36, Ptrend<0.001) and secondary prevention treatments (from 0.46 to 0.70, Ptrend<0.001) were improved. A temporal decrease was found in discharge against medical advice (DAMA) (from 15.2% (95% CI 13.7% to 16.7%) to 8.6% (8.1% to 9.0%); adjusted Ptrend=0.046), and decreases in in-hospital mortality (0.7% in 2015 vs 1.8% in 2005; adjusted OR (aOR) 0.52; 95% CI 0.32 to 0.85) and the composite outcome of in-hospital mortality or DAMA (8.4% in 2015 vs 13.9% in 2005; aOR 0.65; 95% CI 0.47 to 0.89) were observed. Disparities between rural and urban hospitals narrowed; however, disparities persisted in in-hospital management (brain MRI: rural-urban difference from -14.4% to -11.2%; cerebrovascular assessment: from -20.3% to -16.7%; clopidogrel: from -2.1% to -10.3%; anticoagulant for atrial fibrillation: from -10.9% to -8.2%) and in-hospital outcomes (DAMA: from 2.7% to 5.0%; composite outcome of in-hospital mortality or DAMA: from 2.4% to 4.6%). CONCLUSIONS: From 2005 to 2015, improvements in hospital admission and in-hospital management for ischaemic stroke in China were found. A temporal improvement in DAMA and improvements in in-hospital mortality and the composite outcome of in-hospital mortality or DAMA were observed. Disparities between rural and urban hospitals generally narrowed but persisted.

LncRNA RMRP accelerates autophagy-mediated neurons apoptosis through miR-3142/TRIB3 signaling axis in alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a major neurodegenerative disorder. The functions of lncRNA RMRP have been characterized mainly in various human cancers. However, the functional network of RMRP in AD progression remains unknown. METHODS: Human serum samples, AD transgenic (Tg) mice as well as SH-SY5Y cells were used in this study. The RNA expression patterns of RMRP, miR-3142 and TRIB3 were assessed by quantitative real-time PCR (qRT-PCR). Levels of apoptosis- or autophagy-associated biomarkers and TRIB3 level were evaluated using immunohistochemistry (IHC), western blotting or immunofluorescence assays, respectively. Bioinformatics methods and luciferase assays were used to predict and validate the interactions among RMRP, miR-3142, and TRIB3. Flow cytometry, TUNEL staining and EdU assays were used to examine the apoptosis and proliferation of neurons, respectively. RESULTS: The elevated RMRP and TRIB3 expressions and activation of autophagy were observed in AD. Knockdown of RMRP restrained neuronal apoptosis and autophagy activation in vitro and in vivo. Interestingly, TRIB3 overexpression reversed the biological effects of RMRP silencing on Aß1-42-induced cell apoptosis and autophagy. Further mechanistic analysis showed RMRP acted as a sponge of miR-3142 to elevate TRIB3 level. CONCLUSION: These data illustrated that knockdown of RMRP inhibited autophagy and apoptosis via regulating miR-3142/TRIB3 axis in AD, suggesting that inhibition of RMRP maybe a therapeutic strategy for AD.

Correlation between Fibrinogen and White Matter Hyperintensities among Nondiabetic Individuals with Noncardiogenic Ischemic Stroke.

OBJECTIVE: The aim of the present study is to confirm the correlation between fibrinogen and the severity of cerebral white matter hyperintensities (WMHs) among nondiabetic patients with noncardiogenic acute ischemic stroke. PATIENTS AND METHODS: A cross-sectional study of 170 consecutive patients with noncardiogenic acute ischemic stroke who underwent magnetic resonance imaging and vascular imaging was conducted. WMHs were classified into periventricular hyperintensity (PVH) and deep and subcortical WMH (DSWMH) using Fazekas rating scale. After adjustment for fibrinogen and other vascular risk factors, we determined which factors were independent of WMHs. RESULTS: After adjustment for the vascular risk factors, prior ischemic stroke (odds ratio [OR] 4.153, 95% confidence interval [CI] 1.077-16.020, P = .039), fibrinogen level (OR 2.114, 95% CI 1.034-4.322, P = .040), and glycosylated hemoglobin A1c (OR .633, 95% CI .423-.947, P = .026) were independently and positively associated with PVH (P < .05); prior ischemic stroke (OR 2.841, 95% CI 1.469-5.493, P = .002), lipoprotein(a) (OR 1.002, 95% CI 1.000-1.005, P = .047), and fibrinogen levels (OR 1.788, 95% CI 1.170-2.732, P = .007) were independently and positively associated with DSWMH (P < .05). CONCLUSIONS: Our study demonstrated that prior ischemic stroke and higher fibrinogen are associated with WMHs, regardless of PVH and DSWMH, in nondiabetic patients with noncardiogenic acute ischemic stroke. In addition, lipoprotein(a) might be an independent predictor of DSWMH in patients with noncardiogenic acute ischemic stroke.

Correlation between acute stroke-induced white matter lesions and insulin resistance.

The present study was to examine the relationship between white matter lesions (WMLs) and insulin resistance (IR) in patients with acute stroke and evaluate clinical prognosis.Around 200 patients with initial onset of acute stroke including 146 patients (73.0%) with WMLs and 54 patients (27%) without WMLs were analyzed by neuropsychological scales. Fasting blood glucose (FBG), fasting insulin, blood lipid, homocysteine (Hcy), high-sensitivity C-reactive protein (hs-CRP), creatinine, and uric acid, diabetes mellitus (DM), prediabetes (PD), and normal glucose (NG) were determined according to HbA1c levels. According to homeostasis model assessment (HOMA)-IR index of IR, HOMA-IR index ≥2.5 indicated IR, and HOMA-IR index < 2.5 represented noninsulin resistance (NON-IR).IR values and the proportion of patients with IR, HbA1c levels and the quantity of DM patients, the levels of low-density lipoprotein cholesterol, Hcy, and hs-CRP of patients with WMLs were significantly higher than those in patients without WMLs (all P < .05). OR value of IR exposure and WMLs was 1.862 (1.235-2.236). OR values of level 1, level 2, and level 3 WMLs were 1.632 (1.032-2.532), 1.328 (1.152-2.865), and 1.158 (0.639-3.526), respectively. Regarding WMLs patients, MoCA and MMSE scores were significantly decreased, and Hamilton Depression Scale scores were significantly increased (all P < .05). National Institutes of Health Stroke Scale and modified Rankin scale scores of patients with WMLs were significantly increased, and BI scores were significantly decreased (all P < .05).IR is intimately correlated with the WMLs of acute stroke. The incidence and severity of WMLs are significantly associated with cerebral arterial thrombosis, neuropsychology, and neurological scores.

Upregulation of miR-216a exerts neuroprotective effects against ischemic injury through negatively regulating JAK2/STAT3-involved apoptosis and inflammatory pathways.

OBJECTIVE: Ischemic stroke remains a significant cause of death and disability in industrialized nations. Janus tyrosine kinase (JAK) and signal transducer and activator of transcription (STAT) of the JAK2/STAT3 pathway play important roles in the downstream signal pathway regulation of ischemic stroke-related inflammatory neuronal damage. Recently, microRNAs (miRNAs) have emerged as major regulators in cerebral ischemic injury; therefore, the authors aimed to investigate the underlying molecular mechanism between miRNAs and ischemic stroke, which may provide potential therapeutic targets for ischemic stroke. METHODS: The JAK2- and JAK3-related miRNA (miR-135, miR-216a, and miR-433) expression levels were detected by real-time quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blot analysis in both oxygen-glucose deprivation (OGD)-treated primary cultured neuronal cells and mouse brain with middle cerebral artery occlusion (MCAO)-induced ischemic stroke. The miR-135, miR-216a, and miR-433 were determined by bioinformatics analysis that may target JAK2, and miR-216a was further confirmed by 3' untranslated region (3'UTR) dual-luciferase assay. The study further detected cell apoptosis, the level of lactate dehydrogenase, and inflammatory mediators (inducible nitric oxide synthase [iNOS], matrix metalloproteinase-9 [MMP-9], tumor necrosis factor-α [TNF-α], and interleukin-1ß [IL-1ß]) after cells were transfected with miR-NC (miRNA negative control) or miR-216a mimics and subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) damage with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, annexin V-FITC/PI, Western blots, and enzyme-linked immunosorbent assay detection. Furthermore, neurological deficit detection and neurological behavior grading were performed to determine the infarction area and neurological deficits. RESULTS: JAK2 showed its highest level while miR-216a showed its lowest level at day 1 after ischemic reperfusion. However, miR-135 and miR-433 had no obvious change during the process. The luciferase assay data further confirmed that miR-216a can directly target the 3'UTR of JAK2, and overexpression of miR-216a repressed JAK2 protein levels in OGD/R-treated neuronal cells as well as in the MCAO model ischemic region. In addition, overexpression of miR-216a mitigated cell apoptosis both in vitro and in vivo, which was consistent with the effect of knockdown of JAK2. Furthermore, the study found that miR-216a obviously inhibited the inflammatory mediators after OGD/R, including inflammatory enzymes (iNOS and MMP-9) and cytokines (TNF-α and IL-1ß). Upregulating miR-216a levels reduced ischemic infarction and improved neurological deficit. CONCLUSIONS: These findings suggest that upregulation of miR-216a, which targets JAK2, could induce neuroprotection against ischemic injury in vitro and in vivo, which provides a potential therapeutic target for ischemic stroke.

Effects of LRRK2 Inhibitors on Nigrostriatal Dopaminergic Neurotransmission.

INTRODUCTION: Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most prevalent cause of familial and sporadic Parkinson's disease (PD). Because most pathogenic LRRK2 mutations result in enhanced kinase activity, it suggests that LRRK2 inhibitors may serve as a potential treatment for PD. To evaluate whether LRRK2 inhibitors are effective therapies for PD, it is crucial to know whether LRRK2 inhibitors will affect dopaminergic (DAergic) neurotransmission. However, to date, there is no study to investigate the impact of LRRK2 inhibitors on DAergic neurotransmission. AIMS: To address this gap in knowledge, we examined the effects of three types of LRRK2 inhibitors (LRRK2-IN-1, GSK2578215A, and GNE-7915) on dopamine (DA) release in the dorsal striatum using fast-scan cyclic voltammetry and DA neuron firing in the substantia nigra pars compacta (SNpc) using patch clamp in mouse brain slices. RESULTS: We found that LRRK2-IN-1 at a concentration higher than 1 µM causes off-target effects and decreases DA release, whereas GSK2578215A and GNE-7915 do not. All three inhibitors at 1 µM have no effect on DA release and DA neuron firing rate. We have further assessed the effects of the inhibitors in two preclinical LRRK2 mouse models (i.e., BAC transgenic hG2019S and hR1441G) and demonstrated that GNE-7915 enhances DA release and synaptic vesicle mobilization/recycling. CONCLUSION: GNE-7915 can be validated for further therapeutic development for PD.

Genome-wide gene expression analysis of peripheral leukocytes in relation to the male predominance of Guillain-Barre syndrome: differential gene expression between male and female patients.

BACKGROUND: Guillain-Barre syndrome (GBS) fulfils most of the clinical features of an autoimmune disease except for its male predominance. No previous studies have evaluated the differential genome-wide expression between male and female GBS patients. OBJECTIVE: This study sought to identify differences between male and female GBS patients in the gene expression profiles of peripheral leukocytes. METHODS: We downloaded gene chip data-sets pertaining to peripheral leukocyte samples from GBS patients using the gene expression omnibus (submitted by Chang et al.) and applied hierarchical cluster analysis to detect whether there was a gender difference in genome-wide gene expression levels. Then, we identified the sexually differentially expressed genes using a bioinformatic approach and applied enrichment analysis to the gene ontology and Kyoto Encyclopaedia of Genes and Genomes terms to identify significant pathways related to these genes. RESULTS: We observed gender stratification among GBS patients. Twenty genes were expressed more highly in male patients and were enriched for functions, such as macrophage differentiation, leukocyte migration, bladder cancer, pathogenic Escherichia coli infection. In female patients, 62 genes were more highly expressed and were enriched for responses to viral infection and defence, retinoic acid-inducible gene I (RIG-I)-like receptors, cytoplasmic DNA sensing. Matrix metalloproteinase 9 (MMP9) seem to play an important role in the male predominance of GBS. CONCLUSIONS: This study demonstrated gender differences in the genome-wide gene expression of patients with GBS. Bioinformatic approaches offer new means for identifying candidate genes and pathways relevant to the pathophysiology of GBS.

Reduction of α-dystroglycan expression is correlated with poor prognosis in glioma.

Dystroglycan (DG), a multifunctional protein dimer of non-covalently linked α and ß subunits, is best known as an adhesion and transduction molecule linking the cytoskeleton and intracellular signaling pathways to extracellular matrix proteins. Loss of DG binding, possibly by degradation or disturbed glycosylation, has been reported in a variety of cancers. DG is abundant at astroglial endfeet forming the blood-brain barrier (BBB) and glia limitans; so, we examined if loss of expression is associated with glioma. Expression levels of α-DG and ß-DG were assessed by immunohistochemistry in a series of 78 glioma specimens to determine the relationship with tumor grade and possible prognostic significance. α-DG immunostaining was undetectable in 44 of 49 high-grade specimens (89.8%) compared to 15 of 29 low-grade specimens (51.72%) (P<0.05). Moreover, loss of α-DG expression was an independent predictor of shorter disease-free survival (DFS) (hazards ratio (HR) = 0.142, 95% confidence interval (CI) 0.033-0.611, P=0.0088). Reduced expression of both α-DG and ß-DG was also a powerful negative prognostic factor for DFS (HR=2.556, 95% CI 1.403-4.654, P=0.0022) and overall survival (OS) (HR=2.193, 95% CI 1.031-4.666, P=0.0414). Lack of α-DG immunoreactivity is more frequent in high-grade glioma and is an independent predictor of poor clinical outcome. Similarly, lack of both α-DG and ß-DG immunoreactivity is a strong independent predictor of clinical outcome.

Relationship between serum VEGF level and radiosensitivity of patients with nonsmall cell lung cancer among asians: a meta-analysis.

This meta-analysis aims to evaluate the relationships between serum vascular endothelial growth factor (VEGF) level and radiosensitivity in patients with nonsmall cell lung cancer (NSCLC) among Asians. We searched CISCOM, CINAHL, Web of Science, PubMed, Google Scholar, EBSCO, Cochrane Library, and CBM databases from their inception through October 1, 2013. Meta-analysis was performed using the STATA 12.0 software. Fourteen clinical studies were included in this meta-analysis, including five case-control studies and nine cohort studies. Our meta-analysis results revealed that levels of serum VEGF in NSCLC patients were higher than that of healthy controls. There was a significant difference in serum VEGF levels between before and after radiotherapy in NSCLC patients. Further, we found significant differences in serum VEGF levels between effective and noneffective clinical response groups pre- and postradiotherapy. Serum VEGF levels showed no significant associations with tumor-node-metastasis (TNM) stage and histologic grade in NSCLC patients. NSCLC patients with positive VEGF expression had shorter overall survival than those with negative VEGF expression. Our meta-analysis suggests that serum VEGF level may be a useful biomarker in predicting radiosensitivity and prognosis of NSCLC patients among Asians.

Role of Ras, ERK, and Akt in glucocorticoid-induced differentiation of embryonic rat somatotropes in vitro.

This study investigated the roles of Ras, ERK, and Akt in the glucocorticoid-induced differentiation of growth hormone-producing pituitary cells in vitro. Pituicytes isolated from day-18 rat embryos were cultured with 50 mM dexamethasone in addition to specific inhibitors of Ras (manumycin; 0.5, 5, 50 nM), ERK (U0126, 10 µM), or Akt (LY294002, 25 µM). Differentiation was assessed using immunofluorescent staining of intracellular growth hormone. Radioimmunoassay and Western blot analyses were used to determine levels of secreted and intracellular growth hormone, respectively. Manumycin reduced the fraction of growth hormone-positive cells and dexamethasone-induced growth hormone secretion in a dose-dependent manner (both P < 0.001). In the absence of dexamethasone, LY294002 and U0126 did not alter the fraction of growth hormone-positive cells or intracellular growth hormone protein expression or secretion. Both LY294002 and U0126 alone significantly attenuated the fraction of dexamethasone-treated GH-positive cells and the secretion of GH compared to those of cells treated only with dexamethasone (50 nM for 44 h or 48 h) (all P < 0.05). Dexamethasone treatment alone did not change GH protein levels. Treatment of cells with a combination of LY294402 and U0126 significantly attenuated the fraction of dexamethasone-treated GH-positive cells, GH protein levels, and GH secretion compared to cells treated with dexamethasone alone (all P < 0.05). Moreover, dexamethasone-induced phosphorylation of GTP-Ras, ERK, and Akt was significantly attenuated by exposure to the respective inhibitors (P < 0.05). Taken together, our results indicate that Ras, ERK, and Akt are key effectors in the glucocorticoid-induced differentiation of growth hormone-secreting cells.

Therapeutic regimens and prognostic factors of brain metastatic cancers.

OBJECTIVE: This work aims to investigate the therapeutic regimen of brain metastatic cancers and the relationship between clinical features and prognosis. METHODS: Clinical data of 184 patients with brain metastatic cancers were collected and analysed for the relationship between survival time and age, gender, primary diseases, quantity of brain metastatic foci, their position, extra cranial lesions, and therapeutic regimens. RESULTS: The average age of onset was 59.1 years old. The median survival time (MST) was 15.0 months, and the patients with breast cancer as the primary disease had the longest survival time. Females had a longer survival time than males. Patients with meningeal metastasis had extremely short survival time. Those with less than 3 brain metastatic foci survived longer than patients with more than 3. The MST of patients receiving radiotherapy only and the patients receiving chemotherapy only were all 10.0 months while the MST of patients receiving combination therapy was 16.0 months. Multiple COX regression analysis demonstrated that gender, primary diseases, and quantity of brain metastatic foci were independent prognostic factors for brain metastatic cancers. CONCLUSIONS: Chemotherapy is as important as radiotherapy in the treatment of brain metastatic cancer. Combination therapy is the best treatment mode. Male gender, brain metastatic cancers originating in the gastrointestinal tract, more than 3 metastatic foci, and involvement of meninges indicate a worse prognosis.

Poly[tetra-kis(µ(4)-4,6-dimethyl-5-nitro-benzene-1,3-dicarboxyl-ato-κO:O:O:O)bis-(pyridine-κN)dizinc].

In the title complex, [Zn(2)(C(10)H(7)NO(6))(2)(C(5)H(5)N)(2)](n), the repeat unit is a centrosymmetic tetra-carboxyl-ato-O,O'-bridged dimer in which each Zn(II) atom is five-coordinated by four O atoms from different dianionic 4,6-dimethyl-5-nitro-iso-phthalate ligands [Zn-O = 2.0283 (18)-2.0540 (19) Å] and one N atom from a pyridine mol-ecule [Zn-N = 2.030 (2) Å] in the axial site of a slightly distorted square-pyramidal coordination sphere. The Zn⋯Zn separation is 2.9750 (6) Å. The complex dimers are extended into a two-dimensional polymeric structure parallel to (100) through bridges provided by the second carboxyl-ate group of the ligand.

MMP-mediated cleavage of beta-dystroglycan in myelin sheath is involved in autoimmune neuritis.

Alpha-/beta-dystroglycans (DG) located at the outmost layer of myelin sheath play a critical role in its formation and stability in the peripheral nerve system. The demyelination of nerve fibers is present in autoimmune neuritis, however, it is not known about the molecular mechanisms underlying this pathological process. In an animal model of experimental autoimmune neuritis, we observed that beta-DG cleavage was associated with the demyelination of peripheral nerves. The neuritis and beta-DG cleavage were accompanied by matrix metalloproteinase (MMP)-2/-9 over-expressions and attenuated by captopril, a MMP inhibitor. The blockade of MMPs also improves clinical signs. Our results reveal a crucial role of MMP-mediated beta-DG cleavage in autoimmune neuritis, such as Guillain-Barre' syndrome, and bring insights into therapeutic strategies for autoimmune diseases.

Clinical study of arterial anatomic variations for transradial coronary procedure in Chinese population.

BACKGROUND: The radial artery is currently regarded as a useful approach for coronary intervention procedures. Adequate anatomical information of the radial artery should be helpful in performing transradial coronary procedures. Few data about the Chinese population have been obtained in this field. Therefore, we tried to evaluate the incidence and clinical significance of anomalous patterns, and their influence on the intervention procedure. METHODS: In an estimated sample of 3000 cases, radial artery and subclavical artery angiography were performed after insertion of the sheath and coronary angiography (CA). The evaluable data including branch anomaly, tortuosity of the radial artery and procedural characteristics were analyzed. The procedure success was defined as CA or percutaneous coronary intervention (PCI) completed with the initial radial artery approach without changing to other routes. RESULTS: In this study, 1897 cases of CA was undertaken and 1103 cases of CA combined with PCI were performed. The success rate of transradial intervention (TRI) was 96.6% (2899/3000). The approach in 44 cases was changed to the contralateral radial artery and 57 cases were changed to the brachial artery or femoral artery due to failure with the initial radial artery approach. The angiography of the upper limb artery was performed in all cases. Anatomic variations of upper limb arteries were noted in 610 patients (20.3%), which included tortuous configurations of the radial artery (5.0%), hypoplasias (2.2%), radioulnar loop (1.1%), abnormal origin of the radial artery (7.7%), stenosis of radial artery (1.4%), a tortuous configuration of the brachial artery (0.9%), a tortuous configurations of the subclavian artery (1.9%), lusoria subclavian artery (0.1%), and subclavian artery occlusion (0.03%). The procedural success rate in the normal population was higher than in the variation group (97.6% vs 93.0%, P < 0.001). In addition, other procedural outcomes and incidence of complications except radial artery occlusion were also significantly superior to variation group. CONCLUSIONS: Anatomic variations of the radial artery were common, making up an important limitation in the trans-radial approach. Selection of appropriate instruments and understanding some tips and tricks were helpful to overcome the obstacles and effectively reduce the learning curve.

Database searching and accounting of multiplexed precursor and product ion spectra from the data independent analysis of simple and complex peptide mixtures.

A novel database search algorithm is presented for the qualitative identification of proteins over a wide dynamic range, both in simple and complex biological samples. The algorithm has been designed for the analysis of data originating from data independent acquisitions, whereby multiple precursor ions are fragmented simultaneously. Measurements used by the algorithm include retention time, ion intensities, charge state, and accurate masses on both precursor and product ions from LC-MS data. The search algorithm uses an iterative process whereby each iteration incrementally increases the selectivity, specificity, and sensitivity of the overall strategy. Increased specificity is obtained by utilizing a subset database search approach, whereby for each subsequent stage of the search, only those peptides from securely identified proteins are queried. Tentative peptide and protein identifications are ranked and scored by their relative correlation to a number of models of known and empirically derived physicochemical attributes of proteins and peptides. In addition, the algorithm utilizes decoy database techniques for automatically determining the false positive identification rates. The search algorithm has been tested by comparing the search results from a four-protein mixture, the same four-protein mixture spiked into a complex biological background, and a variety of other "system" type protein digest mixtures. The method was validated independently by data dependent methods, while concurrently relying on replication and selectivity. Comparisons were also performed with other commercially and publicly available peptide fragmentation search algorithms. The presented results demonstrate the ability to correctly identify peptides and proteins from data independent acquisition strategies with high sensitivity and specificity. They also illustrate a more comprehensive analysis of the samples studied; providing approximately 20% more protein identifications, compared to a more conventional data directed approach using the same identification criteria, with a concurrent increase in both sequence coverage and the number of modified peptides.

The detection, correlation, and comparison of peptide precursor and product ions from data independent LC-MS with data dependant LC-MS/MS.

The detection, correlation, and comparison of peptide and product ions from a data independent LC-MS acquisition strategy with data dependent LC-MS/MS is described. The data independent mode of acquisition differs from an LC-MS/MS data acquisition since no ion transmission window is applied with the first mass analyzer prior to collision induced disassociation. Alternating the energy applied to the collision cell, between low and elevated energy, on a scan-to-scan basis, provides accurate mass precursor and associated product ion spectra from every ion above the LOD of the mass spectrometer. The method therefore provides a near 100% duty cycle, with an inherent increase in signal intensity due to the fact that both precursor and product ion data are collected on all isotopes of every charge-state across the entire chromatographic peak width. The correlation of product to precursor ions, after deconvolution, is achieved by using reconstructed retention time apices and chromatographic peak shapes. Presented are the results from the comparison of a simple four protein mixture, in the presence and absence of an enzymatically digested protein extract from Escherichia coli. The samples were run in triplicate by both data dependant analysis (DDA) LC-MS/MS and data-independent, alternate scanning LC-MS. The detection and identification of precursor and product ions from the combined DDA search results of the four protein mixture were used for comparison to all other data. Each individual set of data-independent LC-MS data provides a more comprehensive set of detected ions than the combined peptide identifications from the DDA LC-MS/MS experiments. In the presence of the complex E. coli background, over 90% of the monoisotopic masses from the combined LC-MS/MS identifications were detected at the appropriate retention time. Moreover, the fragmentation pattern and number of associated elevated energy product ions in each replicate experiment was found to be very similar to the DDA identifications. In the case of the corresponding individual DDA LC-MS/MS experiment, 43% of the possible detectable peptides of interest were identified. The presented data illustrates that the time-aligned data from data-independent alternate scanning LC-MS experiments is highly comparable to the data obtained via DDA. The obtained information can therefore be effectively and correctly deconvolved to correlate product ions with parent precursor ions. The ability to generate precursor-product ion tables from this information and subsequently identify the correct parent precursor peptide will be illustrated in a companion manuscript.

RAGE expression is up-regulated in human cerebral ischemia and pMCAO rats.

To determine whether the receptor for advanced glycation endproducts (RAGE) contributes to cerebral ischemia, we evaluated RAGE expression in human cerebral ischemia and a model of permanent middle cerebral artery occlusion (pMCAO) in rats. Biopsy specimens were obtained from 12 patients with unilateral cerebral infarction. For the pMCAO model, the middle cerebral artery (MCA) of Sprague-Dawley (SD) rats was permanently occluded. Immunohistochemistry and Western blotting were used to measure RAGE expression in the ischemic hemisphere relative to the normal hemisphere. PC12 cells subjected to oxygen and glucose deprivation (OGD) were used to evaluate the role of RAGE in cell injury. As expected, cerebral ischemia patients expressed elevated levels of RAGE in the ischemic hemisphere. In 1 and 2 days pMCAO rats, levels of RAGE were higher in the ischemic hemisphere relative to the non-ischemic hemisphere, and expression was primarily located in the penumbra of the ischemic hemisphere. In PC12 cells, levels of RAGE increased after 7h of OGD culture. Notably, blockade of RAGE with a selective RAGE antibody in vitro reduced the cytotoxicity caused by OGD. The present data suggest that RAGE is up-regulated in human cerebral ischemia and pMCAO rats, suggesting a role for RAGE in brain ischemia.

[Efficacy of gemcitabine with concurrent radiotherapy on stage III,inoperable non-small cell lung cancer].

BACKGROUND & OBJECTIVE: Radiotherapy is an effective treatment for lung cancer. It is still uncertain whether gemcitabine can improve the efficacy of radiotherapy on lung cancer. This study was to investigate the efficacy of gemcitabine with concurrent radiotherapy on stage III, inoperable non-small cell lung cancer (NSCLC), and observe the adverse events and long-term survival of the patients. METHODS: Sixty patients were enrolled and divided into trial group (30 cases) and control group (30 cases). In trial group, the patients received weekly administration of gemcitabine (400 mg/m(2)) with concurrent radiotherapy at a total dose of 60-66 Gy (2.0 Gy dose fraction per day, 5 days per week). In control group, the patients received only radiotherapy at the same dosage. The responses were evaluated according to WHO criteria. The efficacy, adverse events and long-term survival between the 2 groups were compared with Chi(2) test. The long-term survival was also estimated by Kaplan-Meier method. RESULTS: Fifty-eight patients finished the trial. The follow-up rate was 96.7%. The response rate was 70.0% in trial group and 60.0% in control group (P>0.05). There was no significant difference in the incidence of hematologic and non-hematologic adverse events between the 2 groups (P>0.05). The 1-, 2-, and 3-year survival rates were 77.7%, 58.6%, 26.4% in trial group, and 70.3%, 30.1%, 16.1% in control group (P>0.05). CONCLUSIONS: In this study, gemcitabine with concurrent radiotherapy, as compared with radiotherapy alone, prolonged the survival of stage III, inoperable non-small cell lung cancer patients, but the improvement is not significant. In both groups, the adverse events are tolerable.

[Immunoregulative effect of NK1.1+ cells on the development of experimental autoimmune myasthenia gravis].

AIM: To investigate the effect and immuoregulative mechanisms of NK1.1(+) cells on the development of experimental autoimmune myasthenia gravis (EAMG). METHODS: The NK1.1(+) cells were depleted by intraperitoneal (i.p.) administration of anti-mouse NK1.1 mAb to C57BL/6 mice. Mice were immunized subcutaneously with AChR in CFA. The incidence and severity of EAMG was determined according to the Lennon disease symptoms grading. IFN-gamma and IL-4 in MNCs culture medium were measured by ELISA. AChR IgG of serum was measured by radioimmunoassay. In some experiments, the anti-IFN-gamma mAb was injected (i.p.) to neutralize IFN-gamma. RESULTS: The onset of EAMG was delayed and the severity was decreased obviously in NK1.1(+) cell-depleted mice. However, depletion of NK1.1(+) cells after immunization had no impact on the development of EAMG. Depletion of NK1.1(+) cells could significantly reduce the expression of AChR-specific antibody as well as the production of IFN-gamma. The development of EAMG and production of AChR specific Ab in NK1.1(+) cell-depleted mice were decreased obviously after treating with anti-IFN-gamma antibody. CONCLUSION: NK1.1(+) cells are involved in the early EAMG, and NK1.1(+) cells could enhance the production of IFN-gamma released by AChR-specific T cells as well as the AChR-specific antibodies, which may enhance the outcome of EAMG.