Correlated Secondary Ion Mass Spectrometry-Laser Scanning Confocal Microscopy Imaging for Single Cell-Principles and Applications / 分析化学

Secondary ion mass spectrometry ( SIMS) as a powerful surface analysis technique has been widely applied in semiconductor industry and geology research. Recently, with the development of instrumental technology, SIMS is attracting more and more attention in life sciences. SIMS can provide surface MS spectra, 2D/3D chemical images and depth profiling of substances simultaneously. The minimal lateral resolution of 2D SIMS imaging is 80 to 100 nm, and the longitudinal resolution of 3D SIMS imaging is about 1-5 nm. However, due to lack of specific ions to render the structures of organelles, SIMS imaging for single cells still has great challenges. Optical microscopy, in particular laser scanning confocal microscopy ( LSCM) , has been emerged to be an indispensable technique for single cell imaging and can obtain high spatial 2D/3D imaging to visualize the structures of organelles. Thus, the combinational use of SIMS and LSCM, which takes advantages of SIMS for molecular imaging and LSCM for morphological imaging, has greatly extended the application of SIMS imaging and ensured its accuracy at single cells level, providing novel insights into better understanding of the biological events inside cells. In this review, we focus on the development and application of SIMS imaging and the correlated SIMS and LSCM imaging in the research of cell biology and drug discovery. We anticipate that the combinational use of SIMS and LSCM imaging has promising future in biomedicine and life sciences.

Association of higher resistin levels with inflammatory activation and endothelial dysfunction in patients with essential hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Some studies have shown that serum resistin levels increase in hypertensive patients. Whether the increase of resistin is related to inflammatory or vascular endothelial function is still unknown. We investigated the relationship of increased resistin levels to inflammatory factors and circulating biomarkers of vascular endothelial function in hypertensive patients.</p><p><b>METHODS</b>One hundred and forty-four nondiabetic patients with new onset, hypertension were recruited. Blood pressure, blood glucose, insulin, resistin, tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), von Willebrand factor (vWF), endothelin-1 (ET-1) and nitric oxide (NO) were measured. The homeostasis model assessment, insulin resistance index (HOMA-IR) was calculated. Patients were divided into two groups according to the median level of resistin. Cytokine levels and indicators of vascular endothelial function were compared. Multiple linear regression was used to determine factors influencing resistin.</p><p><b>RESULTS</b>Serum resistin ranged from 2.57 ng/ml to 20.18 ng/ml in hypertensive patients. High resistin group (> 8.36 ng/ml) had higher levels of TNF-α, IL-6, vWF and ET-1 but lower level of NO compared with low resistin group (P < 0.01). Resistin was positively correlated with body mass index, systolic blood pressure, HOMA-IR, low-density lipoprotein cholesterol, TNF-α and ET-1 but negatively correlated with NO (all P < 0.05). Multiple linear regression analysis revealed that HOMA-IR, TNF-α, NO and ET-1 are independent predictors of resistin with standardized regression coefficients of 0.625, 0.368, -0.260 and 0.222, respectively (all P < 0.01).</p><p><b>CONCLUSIONS</b>We conclude that higher resistin levels are associated with inflammatory activation and endothelial dysfunction, because patients with essential hypertension have increased TNF-α, IL-6, vWF and ET-1 and decreased NO. Moreover, the statistical association of resistin with TNF-α, NO and ET-1 suggests involvement of resistin in the progression of hypertension by influencing inflammation and endothelial function.</p>

Impact of mean fasting glucose over the first 72 hours on in-hospital outcomes of patients with ST-segment elevation myocardial infarction / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the impact of mean fasting glucose over the first 72 hours after admission on in-hospital outcomes in patients with ST-segment elevation myocardial infarction (STEMI).</p><p><b>METHODS</b>The data of 357 non-diabetic patients hospitalized with STEMI were collected from the database of Sun Yat-sen Memorial Hospital, affiliated to Sun Yat-sen University between January 2006 and April 2009. The patients were categorized into 3 groups according to mean fasting glucose over the first 72 hours after admission: < 5.6 (n = 165), 5.6 - 7.0 (n = 122) and > 7.0 mmol/L (n = 70). Clinical characteristics, therapeutic approaches and the incidence of heart failure, malignant arrhythmias, and death during hospitalization were compared among groups. Multivariate logistic regression analysis was performed to determine the association between risk factors and in-hospital outcomes. Receiver-operator characteristic (ROC) curve was generated to assess the power of mean fasting glucose on predicting in-hospital death.</p><p><b>RESULTS</b>Age, past history of infarction and early revascularization therapy were similar among groups. Heart rate on admission, white blood cell count, peak CK-MB level, and proportion of extensive anterior infarction were increased in proportion to higher mean fasting glucose levels. Higher mean fasting glucose levels were associated with increased risk of reduced left ventricular ejection fraction, heart failure characterized by higher Killip class, and malignant arrhythmias. After multivariate adjustment, mean fasting glucose remained to be an independent risk factor for increased in-hospital death of patients with STEMI (OR = 1.31, 95%CI: 1.10 - 1.57; P = 0.003). Mean fasting glucose had the higher area under the ROC curve than admission glucose or fasting glucose after admission based on single measurement (0.758, 0.674 and 0.717; P < 0.001).</p><p><b>CONCLUSION</b>Mean fasting glucose during first 72 hours after admission is an independent predictor for in-hospital death and complications in patients with STEMI, which is superior to admission glucose or fasting glucose after admission based on single measurement in predicting in-hospital outcomes.</p>

Regulation of resting intracellular Ca(2+) concentration of ventricular myocytes by beta(3)-adrenoceptor and its signaling pathway in rats with experimental heart failure / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of beta(3)-adrenoceptor (AR) in regulating resting intracellular Ca(2+) concentration of the ventricular myocytes and investigate the signaling pathway in rats with experimental heart failure.</p><p><b>METHODS</b>Rat models of experimental heart failure were established by ligation of the anterior descending artery, and the myocytes were isolated by enzymatic digestion. The resting intracellular Ca(2+) concentration was determined using laser scanning confocal microscopy (LSCM) in the cells stimulated with 1 micromol/L BRL37344 (a selective beta(3)-AR agonist) alone or in combination with PTX, L-NAME, or methylene blue.</p><p><b>RESULTS</b>In the ventricular myocytes from normal control rats, BRL373444 reduced the resting intracellular Ca(2+) concentration of by 45.5%, while the reduction increased to 59.4% in the cells from rats with heart failure. In combination with L-NAME (10 micromol/L), methylene blue (10 micromol/L), and PTX (2 microg/ml), BRL373444 caused a reduction in resting intracellular Ca(2+) concentration of the ventricle myocytes from normal control rats by 10.1%, 16.9%, and 15.4%, respectively in control group, while the rate was 16.9%, 19.3%, and 11.7% in the heart failure group.</p><p><b>CONCLUSIONS</b>Beta(3)-AR agonist can decrease the resting intracellular Ca(2+) concentration of the ventricular myocytes, but the reduction is smaller in cells from rats with heart failure than in cells of normal rats. This effect is mediated through the PTX-NOS-NO pathway.</p>

Clinical analysis of inappropriate shock of implantable cardioverter defibrillators / 中华内科杂志

Objective Implantable cardioverter defibrillator(ICD)can effectively treat lifethreatening ventricular arrhythmias.The most common side effect is inappropriate discharge.This study analyzes the incidence and causes of inappropriate discharges of ICD in our hospital.Methods Forty.threepatients implanted with ICD in our hospital from November 2001 to October 2007 were involved in our study.Patients were followed-up regularly.All episodes recorded and stored in the ICD were analyzed.Results Seven of the 43 patients underwent ninety-six inappropriate discharges.Inappropriate discharges in six patients were caused by supraventricular tachyarrhythmias(SVT).In one patient the discharge was caused by noise.Most inappropriate discharges occurred in the first year after implantation.The history of atrial fibrillation before implantation is an independent predictor of inappropriate discharges.Conclusions The incidence of inappropriate discharge is 16.3%in our study and the most common cause is SVT.Most inappropriate discharges occur in the first year after implantation.Patients with atrial fibrillation history have a higher risk of inappropriate discharges.

Induction of dendritic cells from intra-operative lost blood and their effects on CIK against liver cancer cells in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the practical possibility of inducing dendritic cells (DCs) from mononuclear cells in the lost blood during operation of hepatocellular carcinoma (HCC) patients, and attempted to find a new source of precursor cells for the personalized immunotherapy based on DCs.</p><p><b>METHODS</b>Collected lost blood during hepatectomy from 9 HCC patients and human cord blood from 8 cases of healthy donors undergoing caesarean section. Their mononuclear cells were divided into monocytes and nonadherent lymphocytes. RhGM-CSF and rhIL-4 were administered to induce the monocytes differentiation into DCs, and then loaded with different antigens (lysate antigen of autologous liver cancer cells and cell line SMMC-7721 cells). The lymphocytes were induced into cytokine-induced killer cells (CIK) with IL-2, CD3-Ab, gamma-IFN and PHA. MTT assay was performed to detect the proliferation rate of T lymphocytes mediated by DC and the cytotoxicity of CIK to liver cancer cells.</p><p><b>RESULTS</b>DCs induced from monocytes of the intra-operative lost blood possessed typical morphology and phenotypes. Compared with the DCs from cord blood, the DCs from intra-operative lost blood expressed lower level of surface markers, but both could effectively induce proliferation of CIK and enhance the cytotoxicity of activated CIK against liver cancer cells at similar levels. When the DCs from lost blood and their counterpart from cord blood were both loaded with autologous tumor cell antigen, the proliferation rates of CIK were (388.9 +/- 137.3)% and (315.1 +/- 44.5)%, respectively, and the killing rates against tumor cells were (87.1 +/- 8.0)% and (90.0 +/- 5.1)%, respectively. When the two similar DC groups were loaded with lysate antigen of SMMC-7721 cells, the proliferation rates of CIK were (239.9 +/- 48.7)% and (226.3 +/- 32.3)%, respectively, and the killing rates against tumor cells were (76.4 +/- 7.9)% and (81.1 +/- 4.3)%, respectively. There were no significant differences between those two DC groups. The data also showed that the proliferation and cytotoxicity of CIK induced by DCs loaded with autologous antigen were higher than that of DCs loaded with SMMC-7721 antigen.</p><p><b>CONCLUSION</b>Mononuclear cells separated from intra-operative lost blood of HCC patients can be induced into mature DCs, which can effectively activate CIK and significantly increase its killing effect on the liver cancer cells, and may become a new source of DCs to study and develop vaccines for clinical application.</p>

Resuits of the national surveillance on endemic fluorosis in Fengshun County of Guangdong Province in 2005-2006 / 中国地方病学杂志

Objective To investigated to implement condition of preventive measure and control effect for endemic fluorosis in Fengshun County from 2005 to 2006.Methods It was investigated according to the National Surveillance Program of Endemic Fluorosis.Hupo,Daizai and Anquan Villages of Tangxi Town in Fengshun County were selected as monitoring spots.The usage of reforming water facilities,fluoride content in drinking water and urine of children aged 8-12 years and the prevalence rate of dental fluorosis of children were investigated.Resul tsin 2005 and 2006.a total of 18 reforming water facilities were surveyed and six of which were damaged or out of service.In 2005,the fluoride content in drinking water in the 3 villages was 2.10,1.22 and 0.15 mg/L The prevalence rate of dental fluorosis of children aged 8-12 years was 54.23%(64/118),38.91%(79/203) and 9.10%(6/66).The urine fluoride content of children was 0.95,0.90 and 1.05 mg/L,respectively.In 2006,the fluoride content in drinking water in Hupo,Daizai and Anquan Village was 2.01,1.57 and 0.21 mg/L.The prevalence rate of dental fluorosis of children aged 8-12 years was 26.47%(27/102),12.50%(23/184)and 6.15%(4/65),respectively.The urine fluoride content of children was 0.97,0.61 and 0.59 mg/L.Conclusions The outcome of surveillance data in Fengshun County has reached the sanle level as that of non-disease area.However,the management of reforming water facilities should be improved.

Sodium Channel Current In Hypertrophied Myocardiocytes in Spontaneously hypertensive Rat / 中华高血压杂志

Objective To study the change in sodium channel current(ⅠNa)in the developement of the left ventricular hypertrophy in spontaneously hypertensive rat(SHR).Methods ⅠNa and membrane capacitance(MC)were measured in myocytes of 10,24 and 34-week SHR by the whole-cell patch-clamp technique.Systolic blood pressure(SBP)and left ventricular mass index(LVMI)were detemined.10-week old Wistar rats served as controls.Results LVMI and MC of SHR were significantly higher than those of Wistar rat(P0.05),however the density of ⅠNain 34-week SHR were higher than those of Wistar rat(-18.3?1.9)vs(-15.3?2.0)pA/pF,P

Establishment and evaluation of the diagnostic kit for anti-HIV1/2 antibody and P24 antigen / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish and evaluate an Enzyme Immunoassay diagnostic kit combined with anti-HIV1/2 antibody and P24 antigen for shortening the examination window period of HIV infection in HIV laboratory diagnosis.</p><p><b>METHODS</b>The enzyme-linked reaction plates was coated by anti-HIV P24 monoclonal antibody and HIV 1/2 antigen. Labeling HIV1/2 antigen and anti-HIV P24 polyclonal antibody with horseradish peroxidase, setup an integrated ELISA kit for detecting anti-HIV-1/2 antibody and HIV P24 antigen, and evaluate the specificity and sensitivity of this kit.</p><p><b>RESULTS</b>The sensitivity of testing P24 antigen was up to 0.2 ng/ml. 78 serum samples of patients with AIDS, 85 serum samples of healthy people were compared with Abbott EIA kit, the coincidence was 100%. 12 051 sera from normal persons and patients were examined, the sensitivity of 100 %and specificity of 99.62 %, respectively.</p><p><b>CONCLUSION</b>The anti-HIV1/2 antibody and HIV P24 antigen can be measured at the same time using this EIA kit, while the examination window period of HIV infection is shortened. Thus, the method is suitable for laboratory diagnosis and epidemiological investigation.</p>

Relationship between severity of coronary arterial lesions and pulse pressure in smokers / 中国医师进修杂志

0.05). In the non-smoking group, the pulse pressures of patients with coronary heart disease were significantly higher than those of subjects with negative angiographic results. Among the three subgroups with different severity of coronary arterial lesion, the pulse pressure of 3-vessel diseases was higher than those of both 2-vessel and 1-vessel subgroup. The pulse pressure of subjects with negative angiographic results in smoking group was significantly higher than that in non-smoking group [(56.1?17.2) mm Hg vs (50.9?11.4) mm Hg,1 mm Hg=0.133 kPa P

Effects of FKBP12.6 gene transfection on the contractility of ventricular myocytes of rats with heart failure / 解放军医学杂志

Objective To investigate the effects of FKBP12.6 gene transfection on the contractility of ventricular myocytes of rats with heart failure.Methods Rat model of heart failure was established by a surgical operation of abdominal aortic stenosis.The enzymatic hydrolysis was employed to isolate the rats' ventricular myocytes.Mediated by adenovirus the FKBP12.6 gene was transfected into ventricular myocytes.Ad-GFP infected cardiac myocytes from heart failure rats and normal rats were used as control.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) were used to detect the specific overexpression of FKBP12.6.Transfected ventricular myocytes were loaded with the membrane-permeant Ca2+ dye X-rhod-1-AM.Under the condition of field stimulation,the laser confocal microscope in line-scan and plane-scan mode was used to detect the Ca2+ transients and myocytes contraction.Results The mRNA and protein levels of FKBP12.6 in the myocytes of rats with heart failure were significantly lower than that in normal myocytes(0.47?0.08 vs 0.96?0.12,0.25?0.04 vs 0.48?0.07,P

The changes of intrarenal endothelin system in the course of congestive heart failure induced by rapid right ventricular pacing in dogs / 中国病理生理杂志

AIM: To investigate the changes of intrarenal endothelin system in the course of congestive heart failure. METHODS: A canine congestive heart failure model induced by rapid right ventricular pacing was used in the present study. Twenty-one mongrel dogs divided randomly into 3 groups: control, congestive heart failure 2 weeks(CHF2) and congestive heart failure 4 weeks (CHF4). The severity of heart failure was evaluated by means of hemodynamic measurement. The concentration of plasma endothelin was detected via RIA, and the expression of endothelin was detected by RT-PCR. RESULTS: The concentration of plasma endothelin in both of CHF2 and CHF4 elevated significantly. In CHF2, the expression of endothelin receptor B(ETB) in renal medulla increased significantly. And in CHF4, the expression of preproendothelin, endothelin receptor A(ETA) and ETB increased both in renal cortex and medulla. Furthermore, in cortex, the expression of ETA increased more significantly than ETB, while in medulla, ETB expressed much more than ETA. CONCLUSION: The changes of renal endothelin system expression plays a role in the regulation of water and electrolyte balance during the progress of congestive heart failure.

Effects of FKBP12.6 overexpression on the performance of sarcoplasmic reticulum in ventricular myocytes of rats with heart failure / 中国病理生理杂志

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

Roles of Rho-kinase in rat cardiac fibroblasts proliferation and collagen synthesis induced by angiotensin Ⅱ / 中国病理生理杂志

AIM: To investigate the role of Rho-kinase signal pathway in rat cardiac fibroblasts (CFBs) proliferation and collagen synthesis induced by angiotensinⅡ (AngⅡ). METHODS: CFBs of neonatal Sprague-Dawley (SD) rats were isolated with the method of trypsin digestion and differential anchoring velocity. The CFBs were stimulated with AngⅡto induce fibrosis. Proliferation of CFBs was observed by MTT coloricmetric assay. Synthesis of collagen was detected by the hydroxyproline. The expression of Rho-kinase mRNA was examined using RT-PCR analysis. The extent of phosphorylation of myosin-binding subunit (MBS-P) of myosin phosphatase was quantified by Western blotting analysis, which was used to evaluate the activity of Rho-kinase.RESULTS: (1) Stimulation of neonatal SD rat CFBs with AngⅡ (10-7 mol/L) significantly increased CFBs proliferation and collagen synthesis (P

Expression of endothelin system in pulmonary tissue in a canine pacing-induced congestive heart failure model / 中国病理生理杂志

AIM: To evaluate expression of the components of endothelin system in pulmonary tissue in a canine pacing-induced congestive heart failure model. METHODS: Twenty-one dogs were divided into 3 groups received 2 (pacing 2 group) or 4 weeks (pacing 4 group) rapid right ventricular pacing, respectively, whereas the other group consisted of 7 dogs received sham-operation as control group. Haemodynamic parameters were detected via left and right heart catheterization. Plasma endothelin-1 was determined by means of RIA. In addition, RT-PCR was used to quantify expression of mRNA of components of endothelin system using ?-actin as internal control. One-way ANOVA and linear regression analysis were used for statistical study. RESULTS:Plasma endothelin-1 increased significantly in heart failure animals. The ratio of preproET-1 to ? actin mRNA was significantly increased from 0.14?0.06 in control group to 0.35?0.08 in pacing 2 group and 0.53?0.08 in pacing 4 group ( P