A Pilot Study of Quantitative Loop-mediated Isothermal Amplification-guided Target Therapies for Hospital-acquired Pneumonia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is important to achieve the definitive pathogen identification in hospital-acquired pneumonia (HAP), but the traditional culture results always delay the target antibiotic therapy. We assessed the method called quantitative loop-mediated isothermal amplification (qLAMP) as a new implement for steering of the antibiotic decision-making in HAP.</p><p><b>METHODS</b>Totally, 76 respiratory tract aspiration samples were prospectively collected from 60 HAP patients. DNA was isolated from these samples. Specific DNA fragments for identifying 11 pneumonia-related bacteria were amplified by qLAMP assay. Culture results of these patients were compared with the qLAMP results. Clinical data and treatment strategies were analyzed to evaluate the effects of qLAMP results on clinical data. McNemar test and Fisher's exact test were used for statistical analysis.</p><p><b>RESULTS</b>The detection of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Stenotrophomonas maltophilia, Streptococcus pneumonia, and Acinetobacter baumannii by qLAMP was consistent with sputum culture (P > 0.05). The qLAMP results of 4 samples for Haemophilus influenzae, Legionella pneumophila, or Mycoplasma pneumonia (MP) were inconsistent with culture results; however, clinical data revealed that the qLAMP results were all reliable except 1 MP positive sample due to the lack of specific species identified in the final diagnosis. The improvement of clinical condition was more significant (P < 0.001) in patients with pathogen target-driven therapy based on qLAMP results than those with empirical therapy.</p><p><b>CONCLUSION</b>qLAMP is a more promising method for detection of pathogens in an early, rapid, sensitive, and specific manner than culture.</p>

In vitro simulation on haemodynamics of mural coronary artery / 医用生物力学

Objective To study the effect of myocardial bridge oppression on blood flow, positive pressure, circumferential stress and shear stress of the coronary artery. Methods The original myocardial bridge simulative device was greatly improved to be able to measure multi-hemodynamic parameters, such as normal stress, circumferential stress and shear stress, so as to exactly simulate real blood dynamics environment with the common effect of several stresses, and comprehensively investigate the relationship between hemodynamics and atherosclerosis of mural coronary artery under the combined effects of several stresses. Results The results from the myocardial bridge simulative device indicated that the hemodynamic abnormalities were mainly located in the proximal end of mural coronary artery, and the mean and oscillation values of normal stress at the proximal end were increased by 27.8% and 139%, respectively, showing a significant increase with the intensification of myocardial bridge oppression. Conclusions It is myocardial oppression that causes the hemodynamic abnormity of proximal coronary artery, which is quite important for understanding the hemodynamic mechanism of coronary atherosclerotic diseases and valuable for studying pathological effects and treatments of the myocardial bridge in clinic.

Efficacy of autologous bone marrow-derived cells transfer for patients with chronic ischemic heart disease: a meta-analysis / 中华心血管病杂志

<p><b>OBJECTIVE</b>We aimed to perform a meta-analysis of clinical trials on the efficacy of autologous bone marrow-derived cells (BMCs) transfer for patients with chronic ischemic heart disease.</p><p><b>METHODS</b>We searched MEDLINE, EMBASE, and Cochrane database through September 2009. Eligible studies were randomized controlled trials of autologous BMCs infusion in patients with chronic ischemic heart disease. We gathered information about left ventricular ejection fraction (LVEF), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and death, and did a random-effect meta-analysis to obtain summary effect estimates for outcomes. The pooled analyses were performed and forest plots were generated with RevMan 5.0 software. Heterogeneity was assessed by meta-regression with STATA 10.0 software. Additionally, subgroup analysis was performed to compare the effect of intracoronary BMCs transfer with intramyocardial cell injection on LVEF.</p><p><b>RESULTS</b>Eleven trials with 490 participants were identified. There were 268 patients in BMCs group, and 222 in control group. In control group, the patients received saline injection or autologous plasma injection or no injection. BMCs transfer was performed via intracoronary transfer or intramyocardial injection. Compared with controls, BMCs transfer significantly improved LVEF by 4.63% (95%CI 2.42 to 6.84; P < 0.01). BMCs transfer was also associated with significant reductions in LVEDV (standardized mean difference -0.55, 95%CI -0.94 to -0.17, P = 0.005) and LVESV (standardized mean difference -0.45, 95%CI -0.73 to -0.17, P = 0.002). In addition, BMCs treatment was associated with a significant effect on death (OR 0.42, 95%CI 0.18 to 1.01, P = 0.05). Subgroup analysis indicated that intramyocardial cell injection was preferred due to its more significant improvement of LVEF than intracoronary cell therapy. Meta-regression suggested the existence of a negative association between baseline LVEF and LVEF change.</p><p><b>CONCLUSION</b>BMCs infusion is associated with a significant improvement in LVEF, and an attenuation of left ventricular remodeling.</p>

Systematic review on safety of intracoronary autologous bone marrow stem cells transfer in patients with acute myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To estimate the safety of intracoronary autologous bone marrow stem cells (BMSC) transfer in patients with acute myocardial infarction.</p><p><b>METHODS</b>A systematic literature search of PubMed, MEDLINE, Cochrane EBM, BIOSIS, EMBASE and Chinese Journal Full-text Database between January 1990 and May 2007, was performed. Inclusion criteria required that patients received intracoronary BMSC transfer after coronary reperfusion therapy for primary acute myocardial infarction; study design involved patient randomization and matching placebo group as well as detailed safety data with more than 3 months follow-up results.</p><p><b>RESULTS</b>A total of 5 trials with 620 patients were available for analysis. The pooled statistics showed similar results between BMSC and placebo groups in terms of occurrence of the individual clinical adverse events and the combined endpoint death, recurrence of myocardial infarction, or revascularization procedures. The combined endpoint death, recurrence of myocardial infarction, revascularization procedures, or rehospitalization for heart failure was significantly reduced in the BMSC group compared with the control group at more than one year follow-up (OR = 0.45, 95%CI 0.28 - 0.74, P = 0.002). Likewise, the occurrence of revascularization and the combined endpoint death, recurrence of myocardial infarction, or revascularization procedures were significantly reduced when BMSC transplantation was performed between 4 and 7 days after primary percutaneous coronary intervention (PCI) (OR = 0.60, 95%CI 0.37 - 0.97, P = 0.04; OR = 0.58, 95%CI 0.37 - 0.91, P = 0.02, respectively). In contrast, there was a significant increase in the combined endpoint revascularization and recurrence of myocardial infarction when BMSC transplantation was performed within 24 hours after PCI (OR = 2.56, 95%CI 1.03 - 6.34, P = 0.04).</p><p><b>CONCLUSIONS</b>Post PCI intracoronary autologous BMSC transplantation in patients with acute myocardial infarction is safe, especially in patients received BMSC transplantation between 4 and 7 days after primary PCI than patients received BMSC transplantation within 24 hours post PCI.</p>

Myocardial bridging detection by non-invasive multislice spiral computed tomography: comparison with intravascular ultrasound / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Invasive intravascular ultrasound (IVUS) is current diagnostic standard for myocardial bridging (MB). Non-invasive multislice computerized tomography coronary angiography (MSCT) technique has provided a good anatomical view of the tunnel artery now.</p><p><b>METHODS</b>A total of 51 consecutive patients with atypical or typical angina scheduled for IVUS were enrolled in this study and MSCT was performed 7 days before IVUS. Coronary imaging was quantified using IVUS and MSCT. Four main vessels (left main artery (LMA), left anterior descending (LAD), left circumflex (LCX), right coronary artery (RCA)) were examined.</p><p><b>RESULTS</b>Forty-one out of 51 (80%) patients received metaprolol (25 mg) before the MSCT scan and 25 of them were current beta-blocker users. The mean heart rate was (64 +/- 3) beats per minute. A total of 51 patients underwent IVUS examination (30 with MB and 21 without MB) were chosen for this study. Twenty-eight out of 30 MB cases were correctly diagnosed by MSCT and 2 patients with MB were not detected. Comparison with IVUS, the sensitivity of detection by MSCT was 93%, specificity was 100%. The lumen diameter of the tunnel artery derived from MSCT and IVUS significantly decreased from (2.9 +/- 0.3) mm to (2.4 +/- 0.4) mm (P < 0.001) and from (3.3 +/- 0.3) mm to (2.6 +/- 0.5) mm (P < 0.001), respectively. Minimal and maximal diameters of MB derived from MSCT were significantly smaller than those from IVUS ((2.4 +/- 0.4) mm vs (2.6 +/- 0.5) mm, P < 0.05 and (2.9 +/- 0.3) mm vs (3.3 +/- 0.3) mm, P < 0.05), respectively.</p><p><b>CONCLUSIONS</b>MSCT offers a reliable non-invasive method for MB in LAD and atherosclerosis diagnosis with diagnostic accuracy comparable with invasive IVUS.</p>

Hyperinsulinemia induced immune maturation of human monocyte derived dendritic cells: bridging between diabetes and atherosclerosis / 中华心血管病杂志

<p><b>OBJECTIVE</b>Dendritic cells an hyperinsulinemia are both implicated in the pathogenesis of atherosclerosis. The aim of this study is to explore the effect of high concentration of insulin on the maturation of monocyte-derived dendritic cells (MoDCs) and related signal transduction pathways.</p><p><b>METHODS</b>Human monocytes were purified (over 98%) using Anti-CD14 micro-beads and cultured for 5 days with DC Cellgro medium containing rhGM-CSF (100 microg/L) and rhIL-4 (20 microg/L). Immature DC were then incubated with insulin of various concentrations (0, 1, 10, 100 nmol/L) for 24 hours in the presence or absence of LY294002 (PI3K inhibitor) or PD98059 (MAPK inhibitor). Immunophenotypic expression of CD86 and CD83 were detected using flow cytometry. Endocytosis function of the MoDCs was evaluated using FITC-Dextran and MoDCs secretion IL-12, IFN-gamma and TNF-alpha were measured by ELISA.</p><p><b>RESULTS</b>Insulin induced significantly higher CD83 and CD86 expressions on MoDCs in a dose-dependent manner. The endocytosis function of MoDCs were significantly inhibited and cytokine secretions of IL-12, IFN-gamma and TNF-alpha significantly increased by 10 nmol/L and 100 nmol/L insulin. These effects could be blocked by the LY294002 and PD98059.</p><p><b>CONCLUSION</b>Hyperinsulinemia contributed to atherosclerosis via stimulating immune maturation of MoDCs via both PI3K and MAPK pathways.</p>

Evaluation of myocardial viability with 201Tl/18F-FDG DISA-SPECT technique in patients with acute myocardial infarction after emergent intracoronary autologous bone marrow mononuclear cells transplantation / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the myocardial viability with (201)Tl/(18)F-FDG DISA-SPECT technique in patients with acute myocardial infarction underwent emergent intracoronary autologous bone marrow mononuclear cells (BM-MNC) transplantation.</p><p><b>METHODS</b>Patients with first acute myocardial infarction underwent emergent percutaneous coronary intervention (PCI) were randomized in a 1:1 ratio to either intracoronary transplantation of autologous BM-MNC (n = 20) or to sodium chloride concluding heparin (control, n = 20) via a micro infusion catheter group immediately after PCI. Change in global left ventricular function (LVEF measured by echocardiography) and the myocardial viability detected by (201)Tl/(18)F-FDG DISA-SPECT from baseline and 6-months post transplantation were analyzed.</p><p><b>RESULTS</b>Left ventricular ejection fraction (LVEF) was improved in both groups and the absolute increase (DeltaLVEF) in BM-MNC group was significantly higher than that in control group (7.6% +/- 2.8% vs. 3.0% +/- 2.8%, P < 0.001). In addition, the absolute decrease of myocardial infusion defect detected by (201)Tl SPECT was more significant in BM-MNC group than that in control group (6.7% +/- 3.0% vs. 2.6% +/- 2.6%, P < 0.001) and the number of mismatched segments (indicating viable myocardium) detected by (18)F-FDG SPECT in border zone was also significantly higher in BM-MNC group than that in control group.</p><p><b>CONCLUSION</b>Improved myocardial viability and reduced myocardial infusion defect post emergent intracoronary transplantation of autologous BM-MNC in patients with acute myocardial infarction could be detected by (201)Tl/(18)F-FDG DISA-SPECT technique.</p>

Diagnostic value of multidetector spiral computed tomography in identifying the composition of coronary atherosclerotic plaques / 中华心血管病杂志

<p><b>OBJECTIVE</b>To estimate the feasibility of 16-multidetector spiral computed tomography (16-MDCT) on detecting coronary plaques in comparison with intravascular ultrasound (IVUS).</p><p><b>METHODS</b>Sixty-eight patients suspected of coronary heart diseases were examined by 16-MDCT, quantitative coronary angiography (QCA) and IVUS. Coronary stenosis was defined as lumen diameter reduction (DS) >or= 50%. Hounsfield units (HU) were used to determine different types of plaques: soft plaque (<or= 50 HU), fibrous plaque (50 - 119 HU) and calcified plaque (>or= 120 HU).</p><p><b>RESULTS</b>Compared to QCA, the sensitivity and the specificity for patients with DS >or= 50% were 91.8% (112/122) and 97.8% (556/568) respectively, the positive and negative predictive value were 90.3% (112/124) and 98.2% (556/566) respectively. In 96 plaques evaluated both by 16-MDCT and IVUS, 20 and 21 soft plaques, 37 and 36 fibrous plaques, 39 and 38 calcified plaques were identified by 16-MDCT and IVUS respectively. HU value of soft (11 +/- 36), fibrous (83 +/- 20), and calcified (292 +/- 80) plaques were significantly different (P < 0.05).</p><p><b>CONCLUSIONS</b>Noninvasive 16-MDCT could correctly estimate coronary stenosis and coronary plaques compositions.</p>

Three-dimensional morphological measurements of the utricle, saccule and stapes footplate / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To determine the optimal drill area on the footplate with the 3D measurements of the stapes and the vestibular end organs.</p><p><b>METHODS</b>Four temporal bones were extracted from the fresh cadavers and undecalcified polymer-embedded. After serially sectioning, image processing and the 3D precisely reconstruction, a local Cartesian coordinates was established in which the tympanic surface of the footplate was supposed to be XY plane and the Z coordinate axis passed through the central point of the footplate and was vertical to the XY plane. The configurations of the utricle and saccule were delineated quantitatively, and then any distance between one point on the surface of the footplate and another point on the surface of the utricle or saccule and its orientation can be measured.</p><p><b>RESULTS</b>There was a "V" shaped cleft between the utricle and the saccule. The angle of the" V" shaped cleft was 50.31 +/- 19.90 (17.00 - 68.00) degrees. The apex of the cleft directed anterosuperiorly and approached the footplate center, while beneath the posteroinferior part of the footplate was an open and deep area. The vertical distance from the center point of the footplate to the vestibular end organs was (2.20 +/- 0.548) mm, the maximum of 3.0 mm and the minimum of 1.6 mm.</p><p><b>CONCLUSIONS</b>The posterior and inferior quadrant of the footplate may be the optimal drill area for the fenestra.</p>

Involvement of p53-dependent pathway in the antiproliferative activity of emodin in human smooth muscle cell / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate whether p53 pathway participates in the effect of emodin on vascular smooth muscle cell proliferation.</p><p><b>METHODS</b>The effects of emodin on vascular smooth muscle cell proliferation were evaluated by cell count, senescent-associated beta-galactosidase staining, and annexin V staining. DNA synthesis was determined by (3)H-thymidine corporation, cell cycle was analyzed by FACS, the p53 protein level was measured by Western blot and cDNA expression array technology was used to demonstrate the effect of emodin on the simultaneous expression of a large number of genes in cultured vascular smooth muscle cells.</p><p><b>RESULTS</b>Emodin at 1.6-3.1 microg/ml inhibited VSMC growth, at 6.3-12.5 microg/ml promoted VSMC aging and induced VSMC apoptosis at 25.0 microg/ml 24 hours after exposure. Unscheduled DNA synthesis, which was a sensitive indicator for DNA injury, was observed in VSMC following 24 hours emodin exposure. The mRNA and protein levels of p53 were up-regulated in a concentration-dependent manner. Proliferation/carcinogenesis-related genes were down-regulated and other genes related to cell senescence, apoptosis, and DNA damage/repair were up-regulated in VSMC after exposure to emodin for 24 hours. Emodin readily permeated VSMC membrane and mostly located in the cytoplasm and few of them in the nucleus.</p><p><b>CONCLUSIONS</b>The p53 pathway in VSMC was activated post emodin exposure in a concentration-dependent manner and which might be responsible for the observed antiproliferative effects of emodin in vascular smooth muscle cells.</p>

Effects and mechanism of hyperglycemia on development and maturation and immune function of human monocyte derived dendritic cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>Dendritic cells play an important role in the pathogenesis of atherosclerosis. To explore the effects of hyperglycemia on the maturation and immune function of human monocyte derived dendritic cells (MDCs).</p><p><b>METHODS</b>Immature MDCs were cultured in RPMI1640 medium with either 5.5 mmol/L D-glucose (NG), 25 mmol/L D-glucose (HG) or 5.5 mmol/L D-glucose + 19.5 mmol/L mannitol (HM) in the absence or presence of 30 mmol/L N-acetylcysteine [NAC, a reactive oxygen species inhibitor (ROS)] for 48 hours. FACS was used to investigate the MDCs immunophenotypic expression. Immune function was evaluated by allogeneic mixed T lymphocyte reaction and measurement of cytokine levels from culture supernatants. Intracellular ROS production in MDCs was also measured by 2', 7'-dichlorodihydrofluorescein (DCF, 10 micromol/L) fluorescence using confocal laser-scanning microscopy techniques.</p><p><b>RESULTS</b>Compared with NG and HM treated MDCs, the expression of maturation markers such as CD1a, HLA-DR, CD83, CD86 were significantly upregulated, allogeneic T cells proliferation as well as the cytokines secretions (IL-2, IL-12, IL-10 and IFN-gamma) significantly increased in HG treated MDCs. Intracellular ROS production in MDCs was also significantly increased and all these stimulatory effects of HG could be partially attenuated by NAC.</p><p><b>CONCLUSION</b>High glucose promote the maturation of MDCs and augment their capacity to stimulate T-cell proliferation and cytokine secretions at least in part through enhancing intracellular ROS generation. These stimulating effects of high glucose on MDCs maturation may be one of the mechanisms of accelerated atherosclerosis found in patients with diabetes.</p>

Study on Inducing of Cord Blood Derived Endothelial Progenitor Cells into Endothelial Cells / 实用儿科临床杂志

Objective To induce cord blood derived endothelial progenitor cells(EPCs)into endothelial cells,and investigate the feasibility of these cells as the seed cells of tissue engineering cardiovascular replacement.Methods Mononuclear cells were isolated from fresh cord blood by 6%HES and density gradient centrifugation.Isolated cells were cultured in medium supplemented with vascular endothelial growth factor(VEGF).Attached cells were identied by morphology,immunofluorescence staining and flow cytometry.Results The percentage of mononuclear cells isolated from fresh cord blood was(3.4?2.1)?10~7/mL.The morphology of attached cells changed while being cultured and inducted,from small-sized round cells to spindle-like cells,to a typical cobblestone morphology,and the total number of cells was 10~7.After 7 days of culture,immunofluorescence staining showed that the vWF and VEGFR-2 were expressed.Compared with the original,cell markers CD_133 decreased(3.11%?1.05%) to(0.09%?0.02%),P

Upregulating the expression of angiogenesis-related genes by transplanting autologous mononuclear bone marrow cells into myocardial infarction scar and the periphery / 中华心血管病杂志

<p><b>OBJECTIVE</b>To detect the regulation of angiogenic genes involved in the processes of collateral development.</p><p><b>METHODS</b>Myocardial infarction (MI) scar was induced by cryoinjury in New Zealand rabbits. Four weeks after MI, 24 hours before cell transplantation, bone marrow was aspirated from the right thigh bone and mononuclear bone marrow cells (BMCs) were isolated by Ficoll density gradient centrifugation. Then the mononuclear BMCs (n = 8) or IMDM culture medium (n = 8) were transplanted into infarction scar and the periphery. Four weeks after mononuclear BMCs transplantation, DNA microarray analysis was performed to detect the regulation of angiogenesis-related genes in infarction scar and the periphery. And the differences of angiogenic genes expression were compared among several important growth factors by Western blot.</p><p><b>RESULTS</b>DNA microarray analysis showed the detail regulation of genes involved in the angiogenic processes. There were 15 genes upregulated over 3 times in the infarction scar. In addition, we also found more genes are involved in the process of angiogenesis in its periphery than in the infarction scar (40 genes vs. 15 genes). Western bolt analysis further demonstrated that mononuclear BMCs transplantation was capable of increasing the levels of VEGF, FGF and Angiopoietin-I expression in the infarction scar and its periphery, compared with the control group, P < 0.05.</p><p><b>CONCLUSION</b>These findings indicate that the natural angiogenic processes leading to collateral development are extremely complex, since many kinds of bone marrow-derived growth factors involved in the processes after mononuclear BMCs transplantation into infarction sites.</p>

A simple and rapid method for propagation and purification of the peripheral blood gammadeltaT cells / 中国实验血液学杂志

The purpose of this study was to set up an approach for expansion of the peripheral blood gammadeltaT cells from normal subjects in order to explore the characteristics of gammadeltaT cells. Peripheral blood mononuclear cells (PBMNC) were separated from 5 - 10 ml peripheral blood and stimulated by the low molecular peptide derived from Mycobacterium tuberculosis (MTb-Ag), and expanded in rIL-2-containing medium. The relative amount of gammadeltaT cells were measured by anti TCR gammadelta-PE staining and flow cytometry. The Cytotoxicity were detected by gammadeltaT assay. The results showed that after stimulation and expansion for 10 days, gammadeltaT cells increased to 69.2% of the total PBMNC and demonstrated significant cytotoxicity against K562 cells. In conclusion, this is a simple, rapid and specific method for expansion of peripheral blood gammadeltaT cells in vitro.