Background Herpetic stromal keratitis (HSK) is an immunopathologic eye disorder mediated by CD4+ T lymphocytes.Interleukin-9 (IL-9) is a cytokine linked to the process of many immune inflammatory diseases,but whether IL-9 is involved in the immunopathology of HSK remains unclear.Objective This study was to investigate the expression of IL-9 in murine cornea during the development of HSK and its relationship with the degree of HSK.Methods One hundred and eighty clean BALB/c mice were divided into the normal control group (20 mice) and experimental group (160 mice).HSK models were established by scratching on the surface of the cornea followed by inoculation of 1 × 106 plague forming unit(PFU) herpes simplex virus type 1 (HSV-1) KOS strain.Change of ocular surface was examined under the slit lamp biomicroscopy before and I day,3,5,7,8,10,14,21 days after inoculation of HSV-1.The corneas of mice were collected on the time points mentioned above.The relative expression level of IL-9 mRNA in the cornea of mice was detected by real-time PCR.Immunocytochemical localization of IL-9 protein in murine cornea was viewed by a confocal microscope after preparation of the corneal cryostat sections.All experimental manipulations were undertaken in accordance with the institutional guidelines for the care and use of laboratory animals.Results Punctiform,dendriform or geographic defect in corneal epithelium were seen 3 days and healing 6 days after inoculation of HSV-1.However,edema and opacity of the corneal stroma appeared 7 days and peaked 14 days following the inoculation.Real-time PCR assay showed that very little of IL-9 mRNA (A260/A280) was expressed in the cornea in the mice of the control group.But in 1 day,3,5,7,8,10 and 14 days,the relative level of IL-9 mRNA was 6.37±0.45,5.66±0.53,3.93±0.35,3.62±0.34,3.23±0.18,2.57±0.14,2.19±0.20,with a significant difference among the various time points (F=92.764,P=0.000),and IL-9 mRNA in 1 day,3,5,7,8,10 and 14 days after inoculation was significantly higher than before inoculation (P＜0.05);while no markedly difference was found in IL-9 mRNA expression between the 21-day group after inoculation and the normal control group (P =0.340).IL-9 protein was expressed in the epithelial layer,stromal layer and endothelial cell layer of the corneas,with a stronger green fluorescence in the HSK mice,but no expression of IL-9 protein was seen in the control mice.Conclusions HSK model can be successfully created by corneal scratching and inoculation of HSV-1 KOS strain in BALB/c mouse.IL-9 is involved in the pathogenesis and development of HSK mediated by immunoresponse.

A miniaturized potentiometric label-free immunosensor based on the standard complementary metal-oxide-semiconduction transistor(CMOS) process and micro fabrication technique was developed to monitoring diabetes, which could detect the concentrations of glycated hemoglobin (HbA1c) and hemoglobin. This immunosensor includes a micro field-effect transistor based sensor chip integrated with signal readout circuit and a disposable probe electrode. The micro sensor chip was designed by our lab and fabricated by Chartered Semiconductor, Singapore. The disposable probe electrode, which was integrated with sensitive electrodes array and micro reaction pool, was deposited on polyester plastic based on micro fabrication techniques. Antibody of HbA1c and hemoglobin were immobilized on the electrode based on self assemble monolayer and gold nanoparticles. The characteristics of the electrode during modification were studied by cyclic voltammetry and electrochemical impedance technique. The response characteristic of the immunosensor was detected. HbA1c from 4 to 24 mg/L and hemoglobin from 60 to 180 mg/L can be detected by this immunosensor.

Objective To investigate the change of serum alkaline phosphatase (ALP) level in patients with acute cerebral infarction (ACI) and its clinical significance.Methods A total of 400 patients with ACI (ACI group) and 240 gender-and age-matched healthy controls during the same period were enrolled in this study.Serum ALP,total protein,alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and other indicators of liver function,serum creatinine (Cr),blood urea nitrogen (BUN) and other indexes of renal function were determined.ACI patients were divided into three subgroups according to the largest diameter of infarction slice on the diffusion weighted imaging (DWI):small area group (＜1.5cm),middle area group (1.5-4.0cm) and large area group (＞4.0cm).The degree of neurologic impairment at admission and the degree of nerve function recovery at discharge were assessed by National Institute of Health stroke scale (NHISS) and modified-Rankin Scale (mRS) in all patients.The relationship of serum ALP level with infarction area,NHISS score and mRS was analyzed.Results Serum ALP level was higher in ACI group than in control group [(80.1±24.4)U/L vs.(75.5±21.5)U/L,t=-2.471,P=0.014].Serum ALP level was elevated along with the increase of infarction areas in ACI group [(78.0 ± 23.3)U/L,(79.4±24.9) U/L vs.(87.7±26.0) U/L,F=4.374,P=0.013].The correlation analysis showed that serum ALP level was positively correlated with infarction area (r=0.118,P=0.019) and NHISS score (r=0.114,P=0.023),but no significant relationship was found with mRS (r=0.071,P=0.08).Multivariate regression analysis revealed that patients with high serum ALP level (＞86 U/L) had 1.691-fold higher risk for cerebral infarction than did those with low serum ALP level(＜67 U/L) (OR=1.691,95％CI:1.103～2.594) after adjusting for confounding variables including age,gender,hypertension,diabetic mellitus,heart disease (coronary heart disease and atrial fibrillation),et al.Receiver operating curves (ROC) analysis revealed that the area under the curve of serum ALP level for predicting ACI was 0.557 (95％CI:0.512～0.602,P=0.016),and its sensitivity and specificity were 43.1％ and 67.6 ％ respectively when serum ALP level was 81.5 U/L.Conclusions Serum ALP level is increased in ACI patients,which has positive correlations with cerebral infarction areas and NHISS scores.In the future,maybe,we could assess ACI by monitoring serum ALP levels or improve the prognosis of ACI patients by decreasing serum ALP levels in the future.

Based on the plasmon coupling effect in gold nanoparticles core-satellite nanostructures linked by thymine(T)-rich DNA hybridization and the specific Hg2+-mediated T-Hg2+-T base pair, a novel localized surface plasmon resonance (LSPR) optical fiber sensor was proposed and developed for Hg2+ detection in water.The Hg2+-induced conformational change in T-rich DNA sequence inhibited the DNA hybridization reaction, weakened the plasmon coupling effect and leaded to the change of LSPR resonance wavelength.The concentration of Hg2+ was quantitatively determined by the resonance wavelength redshift.The linear range of Hg2+ detection was about 5-150 nmol/L with LOD about 3.4 nmol/L.The specificity of the sensor was proved great by evaluating the response to other heavy metal ions such as Zn2+, Mg2+, Pb2+ and so on.This sensor was applied in environmental water detection by standard addition method,with the RSD less than 4.8% and recoveries of 94.2%-105.4%.

Objective To observe the effects of nebulized nitric oxide donors on pulmonary and systemic hemodynamics during pulmonary hypertension induced by hypoxia (FiO_2=0.1,for 1 hour)in newborn piglets. Methods Twenty-four anesthetized and mechanically ventilated piglets were randomly divided into four groups:(1)group S (n=6),control group;(2)group C (n=6),receiving saline after hypoxia;(3)group NTG,receiving nebulized nitroglycerin after hypoxia;(4)group SNP,receiving nebulized sodium nitroprusside after hypoxia. Mean arterial pressure (MAP)and mean pulmonary arterial pressure (MPAP)were monitored continuously. Arterial blood gas analysis was determined at baseline,1 hour after hypoxia,termination of nebulization and 0.5 h after nebulization. After nebulization,blood methemoglobin (MetHb)and plasma NO level were measured.Results Acute hypoxia resulted in a significant increase in MPAP. At 1 hour after hypoxia,MPAP in group C,group NTG and group SNP were significantly higher than that in group S(P0.05).After nebulization,there were no significant differences in plasma NO level and blood MetHb level between groups. No rebound pulmonary hypertension was observed after withdrawal of nebulization. Conclusion Both NTG and SNP nebulization could be used as selective pulmonary vasodilators for acute hypoxic pulmonary hypertension.

Objective To explore the experiences of patients with abnormal extubation of PICC tubes.Methods Fifteen patients with abnormal extubation of PICC tubes participated in the semi-structure interviews.Data were analyzed by Nancy's phenomenological procedure.Results The patients experienced complicated mood which combined negative experience and positive experience,with complainant,helpless,worry,fear for negative experience,relieved,secure for positive experience.Conclusions Patients with abnormal extubation possessed negative experience.Suggestions should be given to nursing staff to communicate with patients before the extubation in order to reduce the dispute of nurses and patients.At the same time,we should summarize the reasons and analyze the factors about the abnormal extubation,and take the targeted intervention measures in the clinical work to decrease the incidence rate of PICC extubation.

Based on microfabrication technology and electrochemical modification method, a micro electrochemical sensor for nitrate ( NO-3 ) determination was developed. A micro sensor chip with working electrode and counter electrode was used as the signal convertor of the sensor. The area of the micro working_electrode was only 1 mm2 . As an electrocatalysis sensitive material, copper was electrodeposited onto the working electrode by square_wave pulse current electrodeposition method. The morphologies and components of freshly deposited materials were examined by scanning electron microscopy ( SEM ) and X_ray diffraction ( XRD) to explore key factors that affected the electrocatalytic ability of the deposited copper layer for reducing nitrate ions. The experimental results revealed that under the optimal conditions, the deposited copper layer was macroporous and had a larger effective surface area that could serve as a more effective electrocatalyst in facilitating nitrate reduction. Electrochemical response of the macroporous copper layer was characterized by linear sweep voltammetry in acidic supporting electrolytes ( pH=2 ) . The electroanalytical results showed that the modified microsensor had marked sensitivity for standard nitrate samples within the concentration range from 12. 5 to 3000 μmol/L (in the range of 12. 5-200 μmol/L yielded straight line:y1=-0. 1422x-10. 326, R12=0. 9976, while in the range of 200-3000 μmol/L yielded straight line: y2=-0. 0984x-22. 144, R22=0. 9927) with a detection limit of 2 μmol/L (S/N=3). The developed electrochemical microsensor was also employed for nitrate determination in water samples collected from lakes and rivers near the city of Beijing. The results were in good agreement with the data given by qualified water quality detection institute, with the deviations from 3 . 9% to 15 . 4%.

A rapid LAMP detection method with primers designed on genus-specific region identified in the gyrA gene was established in this assay. All four Campylobacter jejuni from different sources were detected positive and fourteen non Campylobacter bacteria were negative, which shows excellent specificity of the primers. Compared with plate count and PCR method, the LAMP method and the PCR method had equal sensitivity, which were three orders of magnitude higher than plate count. In this assay, we also found out that the treatment of DNase could reduce the dead bacteria DNA effectively. The LAMP detection on chicken indicated relatively good result on detection of Campylobacter jejuni combining with treatment of DNase.

Natural products are important sources for the discovery of anti-tumor drugs. Evodiamine is the main alkaloid component of the traditional Chinese herb Wu-Chu-Yu, and it has weak antitumor activity. In recent years, a number of highly active antitumor candidates have been discovered with a significant progress. This article reviews the research progress of evodiamine-based antitumor drug design strategies, in order to provide reference for the development of new drugs with natural products as leads.

Objective To assess the expression pattern of protease-activated receptor 2 (PAR2) in human keratinocytes and to characterize its biological functions in the regulation of skin barrier.Methods Primary human keratinocytes and human N/TERT keratinocytes were used as the subject of this study.The expression and distribution of PAR2 in the keratinocytes were analyzed by using immunoflorescence staining and Western blot.Two different PAR2 agonists,trypsin and a PAR2-activating peptide (AP),as well as a PAR2-antagonistic peptide (H2N-FSLLRY-COOH) and a control peptide were used to induce the activation of PAR2 in the keratinocytes.Then,a fluorescence-based calcium mobilization assay was performed to evaluate the biological function of PAR2.Data were statistically analyzed by one-factor analysis of variance.Results Under normal culture conditions,PAR2 was weakly expressed in keratinocytes,and the expression was unaffected by culture medium composition or culture duration.Calcium mobilization was induced by trypsin of 50-250 nmol/L and the PAR2-activating peptide in a dose-and time-dependent pattern.The maximal activation of PAR2 was observed in keratinocytes treated with the PAR2 agonist HAN-SLIGKV-COOH of 75-250 μmol/L.The PAR2-antagonistic peptide (H2N-FSLLRY-COOH) obviously suppressed the increase in calcium mobilization induced by trypsin,while the control peptide PAR-RAP showed no inductive effect on the PAR2 activation based on the absence of calcium mobilization.The substrate-induced calcium release was complete within 250 seconds,and peaked at 50 seconds after the initial trypsin or PAR-AP stimulation.Moreover,the activation of PAR2 was accompanied by an increase in ERK phosphorylation and elicitation of MAPK signaling pathway in keratinocytes.Conclusions Human keratinocytes positively express PAR2,which can be activated by trypsin and PAR2-activating peptides,and the activation of PAR2 may influence the physiological function of keratinocytes by inducing intracellular calcium release.