Search

Fluorescence imaging with one-nanometer accuracy (FIONA) is a simple but useful technique for localizing single fluorophores with nanometer precision in the x-y plane. Here a summary of the FIONA technique is reported and examples of research that have been performed using FIONA are briefly described. First, how to set up the required equipment for FIONA experiments, i.e., a total internal reflection fluorescence microscopy (TIRFM), with details on aligning the optics, is described. Then how to carry out a simple FIONA experiment on localizing immobilized Cy3-DNA single molecules using appropriate protocols, followed by the use of FIONA to measure the 36 nm step size of a single truncated myosin Va motor labeled with a quantum dot, is illustrated. Lastly, recent effort to extend the application of FIONA to thick samples is reported. It is shown that, using a water immersion objective and quantum dots soaked deep in sol-gels and rabbit eye corneas (>200 µm), localization precision of 2-3 nm can be achieved.

Background: The clinical implications of emergent HIV drug resistance on samples with low-level viraemia (LLV <1000 copies/ml) remain unclear. We undertook the present analysis to evaluate the impact of emergent HIV drug resistance at LLV on the risk of subsequent virologic failure., Methods: One thousand, nine hundred and sixty-five patients had genotype results at LLV. Risk of virologic failure (≥1000 copies/ml) after LLV was evaluated by Kaplan-Meier analysis and Cox proportional hazards regression. Resistance was assessed using the Stanford algorithm or virtual phenotypes. Patients were grouped into four susceptibility categories ('GSS' or 'vPSS') during LLV, corresponding to the number of 'active' drugs prescribed: <1; 1-1.5; 2-2.5; and ≥3., Results: A total of 1702 patients with follow-up on constant therapy were eligible for analysis. Participants excluded due to changing therapy or loss to follow-up before their next observation had mostly similar characteristics to included participants. There was a 'dose-dependent' increase in the hazard ratio for virologic failure with susceptibility categories at LLV. Compared with a GSS of at least 3, hazard ratios for virologic failure were 1.4 for GSS 2-2.5; 2.0 for GSS 1-1.5; and 3.0 for GSS less than 1 (P < 0.001). Numerous sensitivity analyses confirmed these findings., Conclusion: Our results demonstrate that emergent HIV drug resistance at LLV is strongly associated with subsequent virologic failure. Furthermore, we uncovered a 'dose-dependent' increase in the hazard ratio for virologic failure with decreasing GSS estimated at the time of LLV. On the basis of these findings, we propose that resistance genotyping be encouraged for HIV-infected individuals on antiretroviral therapy experiencing low-level viraemia.

This study was undertaken to evaluate antihyperlipidaemic and in vitro antioxidant activity of ethanolic extract of Flos populi (EFP). The results demonstrated that EFP contains 78.5% flavonoids and 10.4% phenolics, and it exhibited free radical-scavenging activity on 2,2-diphenyl-picrylhydrazyl (IC50 36.40 ± 0.62 μg/mL) and high reducing power (EC50 206.32 ± 1.6l μg/mL) under in vitro chemical assays. And irrespective of prophylactic administration or remedial administration, in high fat diet-induced hyperlipidaemic mice, oral treatment with EFP produced a decrease in the levels of serum total cholesterol, triglycerides, low-density lipoprotein-cholesterol and increase in high-density lipoprotein-cholesterol. In a word, in high fat diet-fed hyperlipidaemic mice, EFP (100, 200 and 400 mg/kg) significantly altered the plasma lipid levels to near normal. These results support a potential effect of EFP in cardiovascular disease.

The ultrasound-assisted extraction of phenolic compounds from Epimedium brevicornu Maxim was modeled using response surface methodology. A Central Composite Design (CCD) was employed to optimize three extraction variables, including ethanol concentration (X 1), extraction time (X 2), and ratio of aqueous ethanol to raw material (X 3), for the achievement of high extraction yield of the phenolic compounds. The optimized conditions are X 1 of 50% (v/v), X 2 of 27.5 min, and X 3 of 250 mL/g. Under these conditions, the experimental yield is 4.29 ± 0.033% (n = 3). The antioxidant activity was evaluated using the DPPH assay and ferric-reducing antioxidant power (FRAP). And it indicates that the phenolic compounds from Epimedium brevicornu Maxim possess significant antioxidant activity. HPLC analysis reveals that the main phenolic compound in the extract product was identified as gallic acid, catechin (Cianidanol), p-hydroxybenzoic acid, vanillic acid, caffeic acid, ferulaic acid, rutin, benzoic acid, and quercetin.

Quantum dots are promising candidates for single molecule imaging due to their exceptional photophysical properties, including their intense brightness and resistance to photobleaching. They are also notorious for their blinking. Here we report a novel way to take advantage of quantum dot blinking to develop an imaging technique in three-dimensions with nanometric resolution. We first applied this method to simulated images of quantum dots and then to quantum dots immobilized on microspheres. We achieved imaging resolutions (fwhm) of 8-17 nm in the x-y plane and 58 nm (on coverslip) or 81 nm (deep in solution) in the z-direction, approximately 3-7 times better than what has been achieved previously with quantum dots. This approach was applied to resolve the 3D distribution of epidermal growth factor receptor (EGFR) molecules at, and inside of, the plasma membrane of resting basal breast cancer cells.

Objective: Examine the short-term (acute) effects of methylphenidate (MPH) on math performance in children with attention-deficit hyperactivity disorder (ADHD) and what factors predict improvement in math performance., Method: One hundred ninety-eight children with ADHD participated in a double-blind, placebo-controlled, randomized crossover MPH trial. Math response to MPH was determined through administration of math problems adjusted to their academic level during the Restricted Academic Situation Scale (RASS). Student t tests were conducted to assess change in math performance with psychostimulants. Correlation between change on the RASS and change on the math performance was also examined. Linear regression was performed to determine predictor variables., Results: Children with ADHD improved significantly in their math with MPH (P < 0.001). The degree of improvement on the RASS (which evaluates motor activity and orientation to task) and on math performance on MPH was highly correlated. A child's age at baseline and Wechsler Individual Achievement Test (WIAT)-Numerical Operations standard scores at baseline accounted for 15% of variances for acute math improvement., Conclusions: MPH improves acute math performance in children with ADHD. Younger children with lower math scores (as assessed by the WIAT) improved most on math scores when given psychostimulants., Clinical Trial Registration Number: NCT00483106.

Doubly doped YAG:Ce3+, Pb2+ phosphor was obtained by doping YAG:Ce3+ phosphor with PbO. Compared with the emission spectra of YAG:Ce3+ phosphor without PbO, the Ce3+ emission band of YAG:Ce3+, Pb2+ shifts to longet wave, which can enhance the red component of spectrum. Meanwhile, the intensity of Ce3+ emission is increased by 10% when the concentration of PbO is 5%, SEM image indicates that PbO may act as a flux. Temperature-dependent emission spectra of YAG: Ce3+, Pb2+ phosphor show a better thermal quenching characteristics than YAG:Ce3+ phosphor.

Objective: To investigate the factors associated with the age of natural menopause and menopausal symptoms in a large population of Chinese middle-aged women., Study Design: In this cross-sectional study, a total of 20,275 women (40-65 years) attending health screening in Jiangsu Province of China were enrolled. A structured questionnaire was used to collect data of demographics, menopausal status, chronic diseases, reproductive history, etc. Also we evaluated the severity of menopausal symptoms by Kupperman menopause index (KMI)., Main Outcome Measure: Menopausal age and scorings of Kupperman menopause index., Results: The overall median age at natural menopause was 50 years. Lower educational level, poor economic status, lower body mass index (BMI), age at menarche less than 14 years, nulliparity and smoking were associated with earlier onset of natural menopause (P<0.05). The most frequently symptoms in postmenopausal women were sexual problems (57.05%), muscle/joint pain (53.29%) and insomnia (51.02%), while fatigue, insomnia and muscle/joint pain were predominant symptoms in pre- and peri-menopausal women. After adjusting for confounding factors, logistic regression analysis revealed that women with poor educational background, low income, divorce, higher BMI, higher parity, smoking and chronic diseases presented higher KMI scores (P<0.05)., Conclusion: The study provided an estimate of median age at natural menopause in Chinese women. The main factors contributing to earlier onset of menopause and severity of menopausal symptoms were lower educational level, poor economic status, and smoking. Thus, this study provides important insights for physicians to prevent and treat menopause related symptoms., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Objective: To explore the significance of assessing asthma control by high-resolution computed tomography (HRCT) and biological markers in induced sputum., Methods: Forty-eight patients with asthma (asthma group) and 10 healthy subjects (control group) were retrospectively analyzed. The asthma patients were divided into 4 groups based on severity: 6 with near-fatal attacks, 12 with severe, 14 with moderate and 16 with mild asthma. These patients received step therapy for 6 months based on the guidelines for the prevention and treatment of asthma. After achieving asthma control or partial control, HRCT, lung function and cytokine levels in induced sputum were measured. The ratio of wall area to total airway area (WA%), the ratio of 2 airway wall thickness to outer diameter (2T/D) and lung densities in both the inspiratory and expiratory phases were measured. Matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinases-1 (TIMP-1), and transformation growth factor-β(1) (TGF-β(1)) levels in the sputum were assessed by enzyme-linked immunosorbent assay., Results: There were significant differences in forced vital capacity and forced expiratory volume in 1 second as the percentage of predicted value (FVC% and FEV(1)%, respectively), the ratio of FEV(1)/FVC, and diffusing capacity of the lung for carbon monoxide (D(LCO)) among groups (F = 5.526, 15.064, 16.326, 2.945, respectively, P < 0.05). Sputum levels of MMP-9, TIMP-1 and TGF-β(1) were significantly increased in the near-fatal asthma, severe asthma, moderate asthma and mild asthma groups [MMP-9: (80 ± 16), (70 ± 9), (59 ± 6), and (52 ± 7) µg/L, respectively; TIMP-1: (212 ± 95), (258 ± 167), (28 ± 51), and 98 ± 60 µg/L, respectively; TGF-β(1): (586 ± 81), (513 ± 54), (401 ± 45) and (351 ± 57) µg/L, respectively]compared with the control group [MMP9: (46 ± 5) µg/L; TIMP: (19 ± 13) µg/L; and TGF-β(1): (258 ± 29) µg/L]. These parameters were progressively increased in the asthma groups with the severity of disease (F = 11.179, 49.914, 9.286, respectively, P < 0.05). The ratio of MMP-9/TIMP-1 in sputum was decreased in the near-fatal attack, severe, moderate and mild asthma groups (0.50 ± 0.28, 0.34 ± 0.13, 0.53 ± 0.22, and 0.87 ± 0.75, respectively) compared with the control group (2.93 ± 1.13). The MMP-9/TIMP-1 ratio in the severe asthma group was lowest among the asthma groups (F = 43.335, P < 0.05). 2T/D and WA% were higher in both the near-fatal asthma group (0.51 ± 0.01 and 0.75 ± 0.01, respectively) and the severe asthma group (0.53 ± 0.03 and 0.77 ± 0.03, respectively) as compared to the moderate asthma group (0.43 ± 0.04 and 0.67 ± 0.04, respectively) or the mild group (0.42 ± 0.04 and 0.66 ± 0.04, respectively). 2T/D and WA% were higher in the asthma groups than in the control group (0.35 ± 0.03 and 0.57 ± 0.04, respectively), (F = 40.224, 41.294, respectively, P < 0.05). Lung densities in both the inspiratory and expiratory phases were lower in the near-fatal attack group as compared to those in the other asthma groups or the control group; and the lung density differences between the two phases in the near-fatal attack group were smaller than those in the other asthma groups or the control group (F = 5.048, 13.247, 11.541, respectively, P < 0.05). 2T/D and WA% were correlated positively with MMP-9, TIMP-1 and TGF-β(1) levels, but negatively with the MMP-9/TIMP-1 ratio, respectively., Conclusions: HRCT and biological markers in induced sputum could be used to accurately evaluate asthma control. These findings suggest that the severity of asthma, especially, near-fatal attack of asthma, is correlated not only with the degree of airway remodeling, but also with the degree of air trapping.

Background: VEGF is a prime mediator of tumorigenesis and metastasis. Various studies assessing the prognostic value of VEGF in patients with esophageal cancer remain controversial. This study aims to comprehensively and quantitatively summarize the evidence on the suitability of VEGF to predict patients' survival., Methods: Searches were applied to PubMed and EMBASE until December 31, 2011, without language restrictions. Studies were assessed for quality using REMARK (Reporting recommendations for tumor MARKer prognostic studies). Data were collected comparing overall survival in patients with high VEGF level with those with low level. We conducted a systematic review of 31 studies (n = 2,387 patients) and completed a meta-analysis of 30 studies (n = 2,345 patients) that correlated VEGF levels with overall survival. Data were synthesized with HRs., Results: The estimated risk of death was 1.82-fold greater in patients with high VEGF expression [95% confidence interval (CI), 1.58-2.08]. The heterogeneity was not significant (P = 0.130) between studies. High VEGF expression was associated with worse survival in esophageal squamous cell carcinoma (HR, 1.81; 95% CI, 1.57-2.10) and there was no significance in between-study heterogeneity (P = 0.185). Data collected were not sufficient to determine the prognostic value of VEGF in patients with esophageal adenocarcinoma., Conclusions: In this meta-analysis, elevated VEGF expression was associated with poor survival in patients with esophageal cancer but not esophageal adenocarcinoma., Impact: These results support further investigation of VEGF expression for predicting poor survival in patients with esophageal carcinoma and may have implications for treatments directed at inhibiting VEGF-mediated angiogenesis., (©2012 AACR)

To measure nanometric features with super-resolution requires that the stage, which holds the sample, be stable to nanometric precision. Herein we introduce a new method that uses conventional equipment, is low cost, and does not require intensive computation. Fiduciary markers of approximately 1 µm x 1 µm x 1 µm in x, y, and z dimensions are placed at regular intervals on the coverslip. These fiduciary markers are easy to put down, are completely stationary with respect to the coverslip, are bio-compatible, and do not interfere with fluorescence or intensity measurements. As the coverslip undergoes drift (or is purposely moved), the x-y center of the fiduciary markers can be readily tracked to 1 nanometer using a Gaussian fit. By focusing the light slightly out-of-focus, the z-axis can also be tracked to < 5 nm for dry samples and <17 nm for wet samples by looking at the diffraction rings. The process of tracking the fiduciary markers does not interfere with visible fluorescence because an infrared light emitting diode (IR-LED) (690 and 850 nm) is used, and the IR-light is separately detected using an inexpensive camera. The resulting motion of the coverslip can then be corrected for, either after-the-fact, or by using active stabilizers, to correct for the motion. We applied this method to watch kinesin walking with ≈ 8 nm steps.

Objective: Crocin has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of crocin against human colon cancer cells in vitro., Methods: Cell proliferation was examined using MTT assay and the cell cycle distribution fractions were analyzed using flow cytometric analysis after propidium iodide staining. Apoptosis was detected using the TUNEL Apoptosis Detection Kit with laser scanning confocal microscope. DNA damage was assessed using the alkaline single-cell gel electrophoresis assay, while expression levels of p53, cdk2, cyclin A and P21 were examined by Western blot analysis., Results: Treatment of SW480 cells with crocetin (0.2, 0.4, 0.8 mmol/L) for 48 h significantly inhibited their proliferation in a concentration-dependent manner. Crocetin (0.8 mmol/L) significantly induced cell cycle arrest through p53-independent mechanisms accompanied by P21 induction. Crocetin (0.8 mmol/L) caused cytotoxicity in the SW480 cells by enhancing apoptosis and decreasing DNA repair capacity in a time-dependent manner., Conclusions: This report provides evidence that crocetin is a potential anticancer agent, which may be used as a chemotherapeutic drug.

The Eu2+ and Dy3+ ion co-doped Sr3Al2O6 phosphor powders with long afterglow were prepared with high temperature solid-state reaction. The phase and the spectra properties of the material were characterized by X-ray diffraction (XRD) and fluorescence spectrophotometer. It was found that the sample is composed of pure Sr3Al2O6 phase. Furthermore, the emission peak of 537 nm under 360 nm excitation and that of 590 nm excited by 468 nm-light were obtained, respectively, and it is more interesting that the emission peaks were at 537 and 590 nm under 394 nm excitation. The effects of different excitation wavelengths on the emission spectrum were explained reasonably by the effect of nephelauxetic effect and crystal field. It revealed that the two types of luminescence with different color were caused by the differences of the center of gravity of the 5d excited state energy level and the split range of 5d energy level.

The structure of the title compound, C(19)H(19)NO(2), contains a seven-membered ring, which is fused to one five- and one six-membered ring, and carries a tolyl substituent. The two benzene rings are oriented relative to each other at a dihedral angle of 86.90 (7)°. In the crystal, mol-ecules are linked by weak inter-molecular C-H⋯O hydrogen bonds.

Objective: To investigate the benefits of long-term therapy with Xuezhikang, a cholestin extract, in combination with calcium channel blockers for improvement of left ventricular (LV) hypertrophy and function in patients with essential hypertension, as determined using echocardiography., Design: Fifty-five (55) hypertensive patients with normal blood low-density lipoprotein cholesterol (LDL-C) levels were randomly assigned to the Xuezhikang group (n = 28, 1200 mg/d of Xuezhikang) or the placebo group (n = 27, matched placebo). All of the patients were treated with extended-release nifedipine (20 mg twice daily). Thirty (30) normotensive subjects, matched for age and gender, were selected as a control group. Conventional echocardiography and tissue Doppler imaging were used to measure the left ventricle (LV) wall thickness and LV diastolic function at weeks 0, 24, and 72 during the period of observation. The serum levels of lipids, carboxy-terminal propeptide of procollagen type I (PIP), and C-reactive protein (CRP) were determined as well., Results: The hypertensive patients had significantly elevated PIP and CRP levels in serum, increased LV wall thickness, and impaired LV diastolic function compared with the normotensive subjects (0.01 < p < 0.05). Compared with the placebo group, the transmitral flow velocities (E/A ratio) (1.11 +/- 0.36 versus 0.85 +/- 0.24, p < 0.01) and the myocardial motion velocities (Em/Am ratio) at the septal mitral annulus (0.90 +/- 0.19 versus 0.70 +/- 0.18, p < 0.05) and the lateral mitral annulus (1.06 +/- 0.20 versus 0.86 +/- 0.14, p < 0.01) were significantly increased, while there was no significant change in the LV wall thickness after 72 weeks of therapy with Xuezhikang. The serum levels of PIP (0.43 +/- 0.13 ng/mL versus 0.51 +/- 0.20 ng/mL, p < 0.05) and CRP (0.32 +/- 0.13 mg/L versus 0.40 +/-0.17 mg/L, p < 0.05) were significantly reduced compared to placebo treatment. There was no significant correlation between changes in LV diastolic function and blood pressure or lipid profile with Xuezhikang therapy., Conclusion: Long-term therapy with Xuezhikang improved LV diastolic function, probably mediated through antifibrotic and anti-inflammatory effects and independent of blood pressure and lipid profiles in patients with essential hypertension.

IgG3 antibody reaction to soluble antigens prepared from schistosomula (SSA), adult worms (SAWA) and eggs (SEA) in laboratory-bred Microtus fortis (Mf), BALB/c mice and Kunming (Km) mice challenged by cercariae of Schistosoma japonicum was detected by indirect ELISA. The effect of purified IgG3 antibody on in vitro killing schistosomula and protecting mice from infection of S. japonicum was evaluated. The IgG3 antibody level in Mf against SSA and SAWA increased significantly by 79.6 percent and 49.6 percent after the fourth week of challenge infection, but no significant increase in BALB/c mice. Purified IgG3 antibody from laboratory-bred Mf and wild Mf effectively killed schistosomula, and that of the wild Mf induced higher worm-reduction rate. The death rate of schistosomula due to IgG3 antibody purified from sera of laboratory-bred Mf and wild Mf was 2.35 and 5.88 times as high as that of Km mice respectively. The results suggest that IgG3 antibody from Microtus fortis may play an important role in immunity against S. japonicum.

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day. Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury (HgT), combinative mercury (HgC) and gaseous mercury (HgG), and content of total organic carbon (TOC), but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average HgT of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with HgC in profile 1992 and 2000, and HgT was significantly correlated with HgG in profile 1996. HgG/HgT ratio in profile samples decreased in the order of (HgG/HgT)1992 > (HgG/Hg)1996 >> (Hg/HgT)2000. A simple outline of Hg release in landfill could be drawn: with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf > root > stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants (e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum) are capable of enriching more Hg than herbaceous plants.

Objective: To investigate whether statins have effect on diastolic function of both left and right ventricles in hypertensive patients., Methods: This is a randomized, mono-blind, placebo-controlled study. 120 systemic hypertensive (HT) patients with normal or slightly elevated cholesterol were randomized to placebo or Xuezhikang (1200 mg/d) for 24 weeks. Extended-release nifedipine was administrated to the HT patients. 30 healthy volunteers served as controls. Plasma were obtained at baseline and 24 weeks after Xuezhikang therapy. Cholesterol and carboxy-terminal peptide of procollagen type I (PIP) were measured. Early diastolic velocity (Em) and late diastolic velocity (Am) were obtained from right atrioventricular ring and left atrioventricular ring with pulsed wave tissue Doppler imaging., Results: The levels of plasma PIP were higher in HT patients. After 24 weeks, the levels of plasma LDL-C, TC and PIP were significantly lower in Xuezhikang group than those in placebo group; Systolic and diastolic pressure were decreased both in placebo group and Xuezhikang group meanwhile pulse pressure was decreased and Em/Am ratio at left atrioventricular ring was higher in Xuezhikang group as compared with those in placebo group., Conclusion: In systemic hypertensive patients, Xuezhikang exerts a beneficial effect on diastolic function of left ventricule via controlling blood pressure, lowering blood lipid and inhibiting myocardial fibrosis.

Objective: To investigate whether xuezhikang has additional beneficial effect on lipid peroxidation, myocardial fibrosis, inflammation and blood pressure control in hypertensive patients without sever hyperlipidemia treated with extended-released nifedipine., Methods: This is a randomized, single-blind, placebo-controlled study, 100 patients with primary hypertension (HT) and normal or lightly elevated cholesterol level were randomized to receive placebo (n=49) or xuezhikang (1200 mg/d, n=51) for 24 weeks on top of extended-released nifedipine (20 mg, bid), 30 healthy volunteers served as controls. Plasma was obtained at baseline and 24 weeks after therapy. Lipids, C-reactive protein (CRP), malondialdehyde (MDA), superoxide dismutase photo-inhibition rate (SOD-PR) and type I collagen carboxypropeptide (PIP) were measured., Results: Normal blood pressure (<140/90 mm Hg) was found in 49 out of 51 xuezhikang treated patients and in 40 out of 49 placebo treated patients (P<0.01) after 24 weeks therapy. Plasma CRP, MDA, SOD-PR and PIP were significantly higher in HT patients than those in normal controls. Plasma CRP, MDA, PIP and SOD-PR were significantly decreased in xuezhikang group while remained unchanged in placebo group after 24 weeks treatment., Conclusion: In patients with primary hypertension without severe hyperlipidemia treated with nifedipine, xuezhikang treatment exerts additional beneficial effects including better blood pressure control, endothelial function improvement, lipid oxidation loading attenuation and anti-inflammation.

In order to probe to the factors for mercury accumulation to natural surface coating, we study the concentration of mercury in Huangpu River surface coatings developed for different depth, different grown days and different seasons and analysis mercury forms of occurrence. The result show the organic matter in biofilm is much high (7.5%-14.5%). Growth time, depth and seasons are all important factors for mercury accumulation to surface coating. Illumination and temperature was stronger and the growth day was longer, The concentration of mercury in biofilm was higher. The mercury forms of occurrence in surface coating were mainly residual matter bound mercury (41.98%) and organic acids bound mercury (27.89%). The percentage of carbonate bound mercury (10.17%) and ion exchangeable mercury (5.33%) was little.

Cytotoxic T lymphocytes (CTLs) specific for the Epstein-Barr virus (EBV) latent membrane protein 2 (LMP2) antigen are important reagents for the treatment of some EBV-associated malignancies, such as EBV-positive Hodgkin's disease and nasopharyngeal carcinoma. However, the therapeutic amount of CTLs is often hampered by the limited supply of antigen-presenting cells. To address this issue, an artificial antigen-presenting cell (aAPC) was made by coating a human leukocyte antigen (HLA)-pLMP2 tetrameric complex, anti-CD28 antibody and CD54 molecule to a cell-sized latex bead, which provided the dual signals required for T cell activation. By co-culture of the HLA-A2-LMP2 bearing aAPC and peripheral blood mononuclear cells from HLA-A2 positive healthy donors, LMP2 antigen-specific CTLs were induced and expanded in vitro. The specificity of the aAPC-induced CTLs was demonstrated by both HLA-A2-LMP2 tetramer staining and cytotoxicity against HLA-A2-LMP2 bearing T2 cell, the cytotoxicity was inhibited by the anti-HLA class I antibody (W6/32). These results showed that LMP2 antigen-specific CTLs could be induced and expanded in vitro by the HLA-A2-LMP2-bearing aAPC. Thus, aAPCs coated with an HLA-pLMP2 complex, anti-CD28 and CD54 might be promising tools for the enrichment of LMP2-specific CTLs for adoptive immunotherapy.

Objective: To develop a diagnostic kit for the detection of anti-Toxoplasma antibodies in human sera by immunoblot technique., Methods: The cytoplasm proteins of Toxoplasma gondii were extracted from collected ascites in Kunming strain mice infected by tachyzoites of Toxoplasma gondii (RH strain). Toxoplasma gondii soluble antigens were separated by SDS-PAGE electrophoresis and transferred to pyroxylin membrane. Using nonpoisonous tetramethylbenzidine (TMB) as horseradish peroxidase substrate for immunoblot, the optimal experimental conditions were selected through comparison of antigen preparation, reagents for blocking or washing, dilution concentration, reaction time, and frequencies of reacting band. Sensitivity, specificity, Youden index and stability were evaluated as the standard for the kit., Results: In 30 sera with anti-Toxoplasma [gM antibodies and 28 with IgG antibodies, the sensitivity for IgM and IgG antibody detection was 90.0% (27/30) and 85.7% (24/28) respectively, the specificity was all 100% in examining 40 healthy control sera, and the Youden index was 0.9 and 0.86 respectively. The kit was stable at 49 degrees C for 6 months., Conclusion: The immunoblot kit shows an easy operation, fast reaction and reliable result, and may be practical.

Aim: To study the effect of HLA-G1 molecule expressed by an endothelial cell line (ECV304) on the cytotoxic activity of allogeneic NK cells., Methods: ECV304 cells were transfected with recombinant plasmid pcDNA3-HLA-G1 by the liposome transfection, and the expressed HLA-G1 on the cell surface was detected by indirect immunofluorescent assay and flow cytometry. The cytotoxic activity of allogeneic NK cells against ECV304 cells was analyzed by the MTT method., Results: HLA-G1 was expressed on the surface of the transfected ECV304 cells. The specific lysis of NK cells against plasmid pcDNA3 transfected ECV304 was (50.6+/-18.1)%, while the specific lysis against pcDNA3-HLA-G1 transfected ECV304 was (29.7+/-11.4)%, which was significantly lower than the former (P<0.001)., Conclusion: HLA-G1 expressed by the ECV304 cells can inhibit cytotoxicity of allogeneic NK cells.

Levels of total mercury, soluble mercury and particle mercury in water of Huangpu River change greatly, their average values are (0.4 +/- 0.44) ng/mL, (0.27 +/- 0.42)ng/mL, (0.13 +/- 0.10) ng/mL respectively. Mercury in water is mainly in the form of soluble mercury. Average mercury content in sediment of Huangpu River is relative high and up to (204.03 +/- 97.41) ng/g, with a range of 70.52 ng/g - 387.30 ng/g. Mercury content is high in the middle reach of Huangpu River, especially in section of Xidu-Nanpu Bridge, and low in upstream and downstream. Distribution of mercury is hightly related with distribution of industry plants and farming. Locations with high mercury content in sediment are in the downstream of locations with high mercury content in water. Mercury (in sediments) is mainly in the form of residue, exchangeable ions, and humics-bound, seldom is in the form of carbonate-bound. Contrary to residue-bound mercury, exchangeable mercury is low in the middle reach, and high in upstream and downstream. There mainly are residue-bound mercury and little humics-bound mercury, exchangeable mercury, and carbonate-bound mercury in sediment in profile, and the residue-bound mercury increases irregularly with depth. Nearing the Mouth of Yangtze River, mercury in sediment becomes more active.

Samples of water, soil, plants, crops and animals collected from Wanshan mercury miner area, Guzhou Province, were analyzed with an AMA-254 liquid/solid mercury analyzer. It was shown that mercury pollution is mainly in soil and air; except water from a pool, no anomaly mercury is found in waters. Mercury content in soil is 24.31-347.52 mg.kg-1, two or three magnitude higher than the average value of Chinese soils; mercury content in plants is 0.47-331.4 mg.kg-1, and moss can enrich much mercury. Due to the long time of exploration and smelting, abundant mercury are accumulated in the local ecosystem, and healthy of local people are threatened seriously.

Human proinsulin C-peptide with C-terminal glutamine amide could be prepared through solid phase method by combining the gamma-carboxyl group of glutamic acid with the amino group of MBHA resin. The protecting groups were cleaved by HF. MBHA resin is relatively inexpensive. The new method is another way for the preparation of human proinsulin C-peptide. The preparation human proinsulin C-peptide of analogue using of PAM resin was also reported.

Objective: To evaluate the procedure to purify IgG antibodies from Microtus fotis serum., Methods: IgG antibodies from sera of three groups of Microtus fotis were purified by protein G or protein A affinity chromatography, their purity and binding capacity were compared., Results: The protein G affinity chromatography was more efficient than protein A affinity chromatography. The antibodies isolated from protein G affinity chromatography showed a higher purity and better activity than that from protein A affinity chromatography monitored by SDS-PAGE and ELISA. The ability of the purified IgG to bind the second antibodies were 8.5 times and 3.1 times that of non-IgG proteins and unpurified sera, respectively., Conclusion: The protein G affinity chromatography is a rapid, convenient and reliable procedure for Microtus fotis serum IgG purification.

The colorectal carcinoma (CRC)-associated CO17-1A/GA733 antigen (Ag) has been the target of a phase II/III randomized trial of passive immunotherapy with monoclonal antibody CO17-1A (Ab1), and phase I active immunotherapy trials with polyclonal anti-idiotypic antibodies (Ab2) mimicking the CO17-1A or GA733 epitope of the Ag. However, monoclonal rat Ab2 BR3E4 directed against Ab1 CO17-1A was superior to polyclonal Ab2 in inducing antigen-specific humoral and cellular immune responses in mice and rabbits. Various forms of Ab2 BR3E4, i.e., BR3E4-F(ab')2 precipitated with aluminum-hydroxide (alum), BR3E4-F(ab')2 coupled to KLH and precipitated or non-precipitated with alum, and BR3E4-IgG in alum or incomplete Freund's adjuvant were compared for their capacity to induce in rabbits anti-anti-idiotypic antibodies (Ab3) that specifically bind to the CO17-1A Ag. BR3E4-F(ab')2 coupled to KLH and precipitated with alum was shown to induce the highest Ab3 titers, followed by Ab2 BR3E4-IgG in alum. Therefore Ab2 BR3E4 as intact IgG (IgG group) or as F(ab')2 coupled to KLH (KLH group), was administered in a phase I trial to 45 patients with CRC, stage Dukes'D (UICC stage IV), with the goal to modulate patients' immune responses to their tumors. Fifteen of 23 patients in the IgG group developed Ab3 binding specifically to Ab2, and in four of these patients the Ab3 also specifically bound to Ag-positive CRC cells. Lymphoproliferative responses to Ab2 and/or GA733-2E Ag stimulation were observed in three of these patients. Eighteen of the 22 KLH group patients tested developed Ab3 and the Ab3 bound specifically to CRC cells in eight patients. Five of the 15 KLH group patients tested developed lymphoproliferative responses to Ab2 and/or GA733-2E Ag. Thus, there was a trend for the KLH group demonstrating higher immune response rates than the IgG group. Clinical responses were rare in these patients with liver metastases.

OBJECTIVE:The purpose of this article is to realize the auto-control of the injection of radiographic drugs in terms of the principle in remote controlling.METHODS:30 cases of sialaden diseases was studied using the auto sialographic instrument by remote controlling,compared to 67 cases studied using sialography by hand-controlling.RESULTS:The auto-sialographic instrument by remote controlling realize the telecontrol of the injection of the sialographic drugs and auto-injection,simplify the working procedure,and prevent medical staffs from X-ray radiation at work.CONCLUSION:The auto sialographic instrument by remote controlling can not only bring much convenience to medical staffs,but also improve the conformation to clinical diagnosis.

Objective: To investigate activation of nuclear factor-kappa B(NF-kappa B) in pulmonary intravascular macrophage (PIM) stimulated with lipopolysaccharide (LPS) and effects of anti-inflammatory drugs dexamethasone (DEX) and aspirin (ASA) on the process., Methods: PIMs isolated from three healthy pigs were cultured and divided into four groups: Control group; LPS- stimulated group; DEX- or ASA-treated group. The NF-kappa B activity of nuclear protein extract from the PIMs and the concentration of tumor necrosis factor alpha (TNF-alpha) in the supernatant were measured by electrophoretic mobility shift assay (EMSA) and radioimmunoassay (RIA), respectively., Results: The activity of NF-kappa B significantly increased from 0.5 hr to 4 hr after LPS stimulation (P < 0.01); The level of TNF-alpha in the supernatant elevated markedly from 1 hr to 2 hr after LPS stimulation (P < 0.01); positive correlation was found between the NF-kappa B activity and the TNF-alpha concentration at 1 hr after LPS stimulation (r = 0.991, P < 0.01). Compared with LPS-stimulated group, both NF-kappa B activity and TNF-alpha concentration were significantly lowered in DEX- or ASA-treated groups (P < 0.01)., Conclusions: LPS might activate NF-kappa B in the PIMs, and induce the increase of transcription and expression of TNF-alpha gene; Both DEX and ASA could inhibit the activation of NF-kappa B and reduce the release of TNF-alpha.