Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.

Objective To explore the distribution of arsenic speciafion and to estimate the effect of arsenic on glutathione(GSH)levels in the blood and liver of mice exposed to different concentrations of inorganic AsⅢ through drinking water.Methods Mice drank water containing arsenite at concentrations of iAsⅢ of 0(contr01),25,50,100 ms/L for 6 weeks.Blood and liver were sampled to asses$the levels of inorganic arsenic(iAs),monomethylarsenic acid(MMA),dimethylarsenic acid(DMA)by the method of hydride generation trapping and ultra-hypothermia coupled with atomic absorption spectrometry,and the level of GSH by the method of 5,5'-Dithio-bis (2-Nitrobenzoic acid).Results Leveh of iAs.MMA and DMA in blood and in liver increased along with the increase of iAs concentrations in drinking water.Primary methylated index(PMI)and secondary methylation index (SMI)of liver and blood were significantly higher in exposed groups than those in control group(P＜0.05).SMI of liver in 50 mg/L exposed group[(50.45±2.94)%]was significantly higher than those in 25 mg/L and 100 mg/Lgroups[(41.68±7.09)%and(41.19±8.87)%,respectively],the difference being statistically significant(P＜0.05).The ratio of iAs.MMA and DMA in blood and liver in exposed group were 2:3:5 and 4:3:3,the percentage of level of organic arsenic(MMA+DMA)were 80%and 60%.GSH in blood and liver in exposed group decreased along with iAs concentrations in drinking water and had significant differences compared with those in control group (P＜0.05).However,levels of GSH in liver and blood did not differ significantly between exposed groups and control group(P＞0.05).Conclusions Membolism of iAs in liver is maximized when the iAs concentrations in drinking water increases to a certain level.However,the percentage of arsenic speciation in blood is different from that in liver,suggesting that other organs and tissues may be capable of methylation of inorganic arsenic.The level of GSH in liver and blood in mice is a good mark tO reflect the toxicity of arsenic.

Objective To investigate the effect of exogenous kallikrein on apoptosis of the neurons aroundthe cerebralinfarctareain rats. Methods Thirty rats wjth cerebral infarction induced by middle cerebral artery occlusion(MCAO)were assigned randomly into 3 groups(n=10),namely the blank control group,saline group,and pAdCMV-HTK group.In the pAdCMV-HTK group,kallikrein gene was delivered into the cerebral ischemie lesion via a replication-defective adenovims using stereotaetic injection technique, and the expression of exogenous kallikrein was detected immunohistoehemically.TUNEL staining was performed to evaluate the neuronal apoptosis around the infarct area,and RT-PCR used to detect the mRNA expressions ofbcl-2,bax and caspase-3 in the brain tissues. Results At 24 h aftertreatment there were some HTK expressed cells found in group C and peal(at 72 h after treatment.While compare with group B and group C,there existed significant difference(112±6.1,68±4.2,59±3.9,P<0.05).At 72 h after treatment,the NSS of group C was significantly lower than that ofgruop B and A(6.70±0.16,8.13±0.16,7.93±0.20,P<0.05);7 days after the treatment,the difference was more significant(5.14±0.18,7.82±0.14,7.91±0.10,P<0.01).Apoptotic cells were mostly seen around the infarct area.The ratsinpAdCMV-HTK group showed significantly reduced number of cells positive for TUNEL staining as compared to those in the saline and blank control groups at 3 days(10.1±0.9,16.7±1.1,and 20.4±0.8,respectively)and 7 days after the treatment(15.2±1.2,33.6±1.3,and 28.8±1.7,respectively)(P<0.05).The mRNA levels ofbc1-2.bax and caspasc-3 were elevated in all the groups at 24 h,peaked at 72 h,and decreased gradually till 7 days alter the treatment.Compared with those in the other two groups，bcl-2 mRNA level in the pAdCMV-HTK group increased slightly P>0.05) while bax and caspase-3 mRNA levels decreased markedly(P<0.05) 72 h and 7 days after the treatment.Conclusion Kallikrein can inhibit neuronal apoptosis around the cerebral infarct and improve the neurological fimction of rats following cerebral infarction probably by reducing the expressions of such apoptotic factors as bax and caspase-3.

Objective To investigate the effects ofkallikrein gene transfer on microvascularproliferation around the cerebral infarct and on the recovery of regional cerebral blood flow (rCBF)following ischemia/reperfusion injury in rats. Methods The rats with cerebral ischemia/reperfusioninjury induced by middle cerebral artery occlusion (MCAO) were randomly assigned into blank controlgroup, saline group, and pAdCMV-HTK treatment group and received corresponding injections into thetissues around the infarct area. Each group was divided into 3 subgroups (n=10) for observation at 12, 24and 72 h after the treatment. The neurological deficits of the rats before and after the treatment wereevaluated using neurological severity scores (NSS), and the expressions of exogenous human tissuekallikrein (HTK) and vascular endothelial growth factor (VEGF) in the brain tissues were detectedimmunohistochemically. TIC staining was performed to measure the changes in the infarct size.14C-iodoantipyrine tracing technique was used to define the rCBF in the rats. Results Compared tothe blank control group, the cerebral infarct size was significantly reduced in pAdCMV-HTK group 24 hafter the treatment, and was further reduced at 72 h (P<0.05). At 24 h after the treatment, the NSS inpAdCMV-HTK group was significantly lower than that in the blank euntrol and saline groups (P<0.05),and was further reduced at 72 h (P<0.01). After MCAO, the VEGF-positive cells were found mostly inthe cortex and the white matter around the infarct area. The expression of VEGF in pAdCMV-HTK groupwas markedly higher than that in the other two groups at 12, 24, and 72 h after the treatment (P<0.05). Inall the 3 groups, the rCBF around the infarct was slightly decreased as compared to that in thecontralateral hemisphere, pAdCMV-HTK slightly increased the rCBF 12 h after the injection (P>0.05),and significant increase in the rCBF occurred 24 h and 72 h after the injection (P<0.05). ConclusionKallikrein gene transfer following cerebral ischemia/reperfusion injury promotes vascular proliferationaround the infarct and increases the rCBF to reduce the infarct volume and attenuate neurological deficitsin rats.

Objective To summarize our experience with intraoperative neural electrophysiological monitoring during spinal cord surgery. Methods The clinical data of 11 patients undergoing spinal cord surgery with intraoperative neural electrophysioiogical monitoring were retrospectively reviewed, and the monitoring was performed by recording the motor-evoked potential (MEP), somatosensory evoked potential (SEP), and evoked electromyography (EMG). Results Subtotal resection of the intramedullary cystic lesion was performed in 1 case and partial resection of the intramedullary tumor in another. In 9 cases of tethered spinal cord syndrome, obvious improvement was obtained in 8 cases, and the other 1 case showed no obvious changes in the symptoms after the operation. In all the 11 cases, the spinal cord remained intact and its function was totally preserved without damage of the eonus medullaris or the cauda equine. Conclusion Combined monitoring of MEP, SEP, and evoked EMG during spinal cord surgery is useful for protecting the spinal cord and the nerves roots, and may enhance the detection of the tethered tissue and ensure better safety of operations.

Objective To explore the regularity of time-dependent changes in morphology and biomechanical properties of brain tissues in pigs,and value the feasibility of deducing the postmortem interval (PMI).Methods Brain tissues were taken from 42 pigs and kept in an artificial climate chamber with the temperature of 25 ℃ and humidity of 75％.The samples were collected from telencephalon at sequential time intervals (0,12,24,36,48,60 h;n =6) according to the principle of predefined time,position,direction,ratio,quantity and shape.The samples fixed with formaldehyde were then immediately tested by mechanical testing machine to obtain their biomechanical parameters and the histological sections were prepared.Results With the extension of PMI (0-60 h),brain tissues gradually became discolored,weak,mudding and liquefied under the influence of autolysis and putrefaction.Both clearance area of the white matter and its integrated optical density (IOD) significantly increased during 0-48 h.Biomechanical properties of brain tissues including the limit load,average force,elastic modulus and fracture energy all presented a declining tendency at the interval of 12-60 h.The limit load was considered highly statistically significant,and statistical differences were found in average force,elastic modulus and fracture energy.Conclusions There exists a significantly negative structure-activity relationship between the morphology of brain tissues and biomechanical properties.The limit load of postmortem brain tissues in 60 h is the optimum in the window period,which can be used as a new method for estimating PMI.

Objective To identify the impact of age and gender on cardiac structure and left ventricular function in normal Chinese by echocardiography. Methods Cardiac structure, valve flow velocity and cardiac function were measured by echocardiography in 15 692 healthy volunteers. Subjects were grouped by age at 5 years interval in population older than 5 years. Children under 5 years were divided into 3 age groups(<1 years,1-3 years,4-5 years). Hierarchical cluster analyses were performed for ages, based on indexes of cardiac structure and function respectively. Results Six groups (< 1 years, 1-3 years,4-5 years, 6-10 years, 11-20 years, ≥21 years) were generated after the age hierarchical cluster analyses based on index of cardiac structure. Four groups (≤30 years, 31-50 years, 51-80 years,≥81 years) were generated based on spectral current flow. Six groups (< 1 years,1-3 years,4-5 years, 6-10 years, 11-15 years, ≥16 years) were generated baaed on left ventricular systolic function and five groups (≤15 years, 16-30 years, 31-50 years, 51-80 years, ≥81 years) were generated based on left ventricular diastolic function. Cardiac structure index were similar between male and female in age groups ≤ 10 years and significantly lower in females than males in age groups ≥ 11 years (P < 0.05). Valve flow velocity was similar between male and female in various age groups (P >0.05). Left ventricular systolic function was similar between male and female in age groups ≤10 years but was significantly higher in males than females in age groups ≥11 years(all P <0.05). Left ventricular diastolic function was similar between female and male in various age groups (P > 0.05) and equally decreased with aging in both female and male subjects. Conclusions The cardiac development in Chinese population can be divided in 6 phases and becomes stable in subjects older than 21 years, left ventricular systolic function becomes stable in subjects older than 16 years and the left ventricular diastolic function declines physiologically with aging.

Objective:To investigate the technique and efficacy of PTES for treatment of L5/S1 disc herniation.Methods:PTES was performed on 52 cases of L5/S1 herniations without spinal instability and central spinal canal stenosis,including 24 cases of high iliac crest,from November 2012 to April 2013.The operation duration,frequency of intraoperative fluoroscopy,blood loss and hospitalization days were recorded.Leg pain was evaluated by using the visual analog scale(VAS)Preoperatively and immediately,1 week,1 month,2 months,3 months,6 months,1 year and 2 years after surgery.The results were determined to be excellent,good,fair,or poor according to the Macnab classification,and complications were observed at 2-year follow-up.Objective:The mean operation duration was(56.3 ±11.5)min per segment.The median frequency of intraoperatively fluoroscopy was 5(3-14)times.The median blood loss was 5(2-20)mL.The median hospital stay was 3(2-4)days.The average postoperative follow-up was(26.2±2.0)months.The median preoperative VAS score of leg pain was 9(6-10),1(0-3)immediately after the operation and 0(0-3)2 years after operation,and the differences were statistically significant(P<0.001).There were 3 cases of lower limb rebound pain 1 week after operation,which were relieved within 2 months after operation.The rate of excellent and good curative effect was 98.1％(51/52)2 years after operation.No complications such as nerve injury,infection,abdominal organ damage and rupture of large vessels occurred.No recurrence occurred.Conclusions:PTES for L5/S1 disc herniation including the cases with high iliac crest is an easy,effective and safe technique.The method has the advantages of simple positioning,easy puncture,simple steps and less fluoroscopy,and the learning curve is not steep for surgeons.

Objective To analyze the incidence and cause of complications during and after interventional therapy for congenital heart disease (CHD). Methods From April 1986 to April 2009, 388 out of 6029 patients with CHD developed complications during and post interventional therapy, another 5 patients died post procedure, clinical data from these 393 patients were retrospectively analyzed. The patients with severe functional insufficiency requiring intervention or surgery during and after interventional therapy were classified as severe complications. Results The overall complication rate was 6. 44% [7.69% post atrial septal defect occlusion, 4.20% post patent ductus arteriosus (PDA) occlusion, 1.31% post percutaneous balloon pulmonary valvuloplasty, 14.94% post veatricular septal defect occlusion, 3.13% post percutaneous closure of aortopulmonary collaterals, 30.95% post catheter embolotherapy of pulmonary arteriovenous malformations, 12.50% post transcatheter closure of coronary artery fistulae, 20.00% post transcatheter closure of ruptured sinus of Valsava aneurysm, 66. 67% post percutaneous balloon aortic valvuloplasty]. The severe complication rate was 0.65% (39/6029). The procedure-related mortality rate was 0.08% (5/6029), 0.26% (2/761) post percutaneous balloon pulmonary valvuloplasty, 0.05% (1/2070)post PDA occlusion, 9.10% (1/11) post balloon atrial septostomy, 33.33% (1/3) post percutaneous balloon aortic valvuloplasty. Emergency Cardiovascular surgery rate was 0.22% (13/6029). Selective surgery was required in 0.13% (8/6029)of patients post procedure. Two patients (0.03%) received permanent pacemaker implantation. Conclusions The severe complications and mortality rate of interventional therapy for CHD are relative low. Post procedure follow-up is needed fro monitoring possible procedure-related complications.

Objective To evaluate the predictive value of admission plasma amino-terminal pro-B type natriuretic peptide(NT-proBNP)on in-hospital mortality in patients with decompensated heart failure.Methods Plasma NT-proBNP levels were measured in patients with decompensated heart failure within 24 hours after admission with ELISA method.The NT-proBNP levels were compared between survivals and dying patients in hospital.ROC analyses were performed to evaluate the predictive value of admission plasma NT-proBNP on in-hospital mortality and to identify the optimal NT-proBNP cut-point for predicting in hospital mortality.A binary logistic regress analyses was used to evaluate if NT-proBNP was an independent predictor for in-hospital mortality.Results A total of 804 patients with decompensated heart failure were enrolled in his study(293 valvular heart diseases,219 ischemic cardiomyopathy,141 dilated cardiomyopathy,14 hypertrophic cardiomyopathy,21 restrictive cardiomyopathy,39 hypertensive heart disease,41 chronic pulmonary heart disease and 36 adult congenital heart disease)and 96 patients were in class Ⅱ,450 in classⅢand 258 in cases Ⅳ according to NYHA Classification.During hospitalization,64 deaths were recorded and the on admission plasma NT-proBNP levels of patients died during hospitalization were significantly higher than those of survivals[4321.1(3063.8,6606.5)pmol/L VS.1921.6(873.9,3739.2)pmol/L,P＜0.01].Area under receiver operating characteristic curve(AUC)of NT-proBNP to predict in-hospital death was 0.772(95%CI:0.718-0.825,P＜0.01),the optimal plasma NT-proBNP cut-point for predicting in-hospital mortality Was 3500 pmol/L,with a sensitivity of 70.3%,a specificity of 72.0%,an accuracy of 71.9%.a positive predictive value of 17.8%and a negative predictive value of 96.6%.Patients whose NT-proBNP levels were equal or more than 3500 pmol/L had a much higher in hospital mortality(17.8%)compared with those with NT-proBNP levels of less than 3500 pmol/L (3.4%),P＜0.01.Binary logistic regress analyses demonstrated that admission plasma NT-proBNP,pneumonia,heart rate and NYHA class were independent predictors for in-hospital mortality in patients with decompensated heart failure(P＜0.05 or 0.01)and admission plasma NT-proBNP Was the strongest predictor for in-hospital mortality.Conclusions Admission plasma NT-proBNP level was an independent predictor for in-hospital mortality in patients with decompensated heart failure.The optimal NT-proBNP cut point for predicting in-hospital mortality was 3500 pmol/L in this patient cohort.