Objective: To establish an enzyme linked immunosorbent assay(ELISA)for erythropoietin(EPO) in serum, and observe its clinical application value thereof. Methods: Prepare the EPO polyclonal antibody, wash the plate with isopropyl alcohol, and then choose the suitable concentration of the antibody, enzyme labeled antibody, and antigen. After the reaction, check the sensitivity, recovery, specificity and stability of the method. The serum samples of anaemia and breast carcinoma individuals were detected. The results of radioimmunodetection were compared with that of normal control group. Results: The immo-assay plate showed strong adherence to proteins. The optimal concentrations of the antibody, enzyme labelled antibody and antigen were 1∶1 000, 1∶6 000 and 1∶800 separately. The sensitivity was 0.46 U/L. The cross-reaction with growth hormone and ferritin was low. The mean recoveries of samples with high and low concentrations were 96.3%, 97.3% respectively. The coefficients of variation of intra-assay and inter-assay were just 8.31% and 7.82%, and the stability was good. The EPO levels were higher in anaemia and breast carcinoma groups than that of normal group. There was no significant difference between the results of the radioimmunodetection and ELISA. Conclusion: The double-antibody sandwich ELISA method was established for EPO in serum, which had certain clinical application value.

Objective To analyze the validity and safety of using a novel posteromedial approach whose L-shaped incision is reversed in treatment of posterior fractures of the tibial plateau. Methotis Anatomical research was performed on 4 fresh-frozen cadaveric specimens with 8 normal knees. All specimens were dissected by the posteromedial reversed L-shaped approach. Parameters of the important structures were measured. Results two important structures were involved in the novel approach. Including medial saral artery and vein and inferior medial genicular artery. The average distance between the eruptive site of medial sural artery and articular surface was 45.29 mm, and the average length ofmedial sural artery was 36.28 mm. The average distance between the eruptive site of inferior medial genicular artery and articular surface was 10.12 mm. The main posterior structures of the proximal tibia were all surveyed by the single approach, including the posterior bi-condyles and the tibial insertion of the posterior crueiate ligament. Conclusion The posteromedial reversed L-shaped approach is an effective approach for treating posterior fractures of the tibial plateau or posterior injuries of the proximal tibia, because it is less invasive, highly safe, simple in anatomy, direct and sufficient in exposure.

Objective To study the impact of the peripheral neurotomy method combined with rehabilitation exercise on gross motor function and balance function of children with cerebral palsy.Methods Matched pair design was used in the study.Thirty children with spastic cerebral palsy hospitalized from May 2011 to Nov.2013 suffering from peripheral neurotomy were assigned as study group,who were conscious and treated by peripheral neurotomy com bined with rehabilitation exercise.In the meanwhile,30 children inpatients at the same time were employed as the control group,who were similar in age and symptoms,with the same gender and the same Gross motor function classification (GMFCS) grade,and the controls were treated only with ordinary rehabilitation exercise.Modified Ashworth score,Gross Motor Function Measure (GMFM) D and E regions scores,physicians rating scale score in the 2 groups were recorded after treatment for 2,4,6,8,10 weeks.Repeated measure analysis of variance method was introduced to analyze the differences between the 2 groups in improved Ashworth muscle tension,gait improvement,gross motor function and balance function.Results The study group was superior over the control group in muscle tension reduction (F =8.177,P =0.006) and gait improvement(F =24.284,P =0.000).The 2 groups were not different statistically in D region evaluation of GMEM (F =0.072,P =0.790) and E region evaluation of GMFM (F =0.000,P =0.985) ; For Berg balance scale,the 2 groups had also no difference(F =0.150,P =0.700).Conclusions Both peripheral neurotomy method combined with rehabilitation exercise and the ordinary rehabilitation exercise method both could improve the gross function and the balance function.Furthermore,the former was superior over the latter in muscle tension reduction and gait improvement.

Objective To analyze the potential factors facilitating post-pyloric placement of spiral naso-jejunum tube in critically ill patients.Methods A retrospective study was carried out in patients requiring enteral nutrition (EN) from Apr 2005 through Dec 2011 in Intensive Care Unit (ICU).Severity of illness was assessed with APACHE Ⅱ score (acute physiology and chronic health evaluation Ⅱ).A selfpropelled spiral naso-jejunum tube was placed and observed for 24 hours.The forward movement and place of the tube tip was checked by bedside X-ray.The APACHE Ⅱ score,therapeutic measures,agents administered within 24 hours after tube insertion were recorded.The patients were divided into the success group and the failure group identified by bedside X-ray whether the tube tip entered into jejunum or not.Univariate analysis and multivariate Logistic regression analysis were used to find out the potential factors impacting on the success or failure in post-pyloric placement of naso-jejunum tube.Results A total of 508 patients composed of 337 male and 171 female,and aged (62.0 ± 19.2) years with APACHE Ⅱ score of (21.9 ± 7.3) were enrolled for study.The placement was successful in 205 (40.4％) of 508 patients.Univariate analysis showed that APACHE Ⅱ score ≥ 20,sedatives and analgesics,catecholamines,prokinetics,artificial airway and mechanical ventilation were potential factors facilitating the post-pyloric placement of naso-jejunum tube.Multivariate logistic regression identified that APACHE Ⅱ score ≥ 20,sedatives and analgesics and prokinetics were independent factors facilitating the post-pyloric placement of naso-jejunum tube.Conclusions The success rate of self-propelled spiral nasojejunal tubes insertion was relatively low.The prokinetics contributed higher success rate of naso-jejunum tube placement than factors of APACHE Ⅱ score ≥ 20,sedative and analgesic,catecholamine drugs,artificial airway and mechanical ventilation.There were no effects of age and gender on the placement of naso-jejunum tube.

onary artery was turtuous and dilated in 4 cases. Conclusion 64 MSCT is emerging as an essential imaging tool for detecting early anomalous origin of the left coronary artery from the pulmonary artery with high resolution and significance.

Tramadol and its metabolites in rat urine were identified by LC-MS(n). Rat urine samples of 0-36 h were collected after ip 9.0 mg x kg(-1) tramadol, then the samples were enriched and purified through solid-phase extraction cartridge. Purified samples were analyzed by LC-MS(n). Possible metabolites were discovered by comparing the full scan and SIM chromatograms of the test samples with the corresponding blanks and analyzing the retention time, quasi-molecular ion and fragment ion of all chromatograms. Nine phase I metabolites and four phase II metabolites were identified in rat urine. One of the metabolites was found first time in living body. The metabolites were formed via the following metabolic pathways: O-demethylation, N-demethylation, hydroxylation, N-oxidation and conjugation. The method can be used to identify tramadol and its metabolites in other animals and human.

Objective To evaluate engraftment status of patients after allogeneic hematopoietic stem cell transplantation(Allo-HSCT) and prompt relapse of disease based on DNA polymorphism analysis technique.Methods Sixty-six cases were detected by DNA polymorphism analysis technique and 25 cases were monitored and analyzed dynamically during this period.Results After Allo-HSCT,48 patients obtained type of donors,but 13 patients did not; 5 patients showed mixed chimerism.Two cases of type of donors converted into mixed chimerism and 4 cases of mixed chimerism converted into type of donors after some time. The others' engraftment status did not change.Conclusion DNA polymorphism analysis technique can detect engraftment status of patients exactly, rapidly, which provides effective evidences of constitution for more clinical therapy projects.

An analytical method of ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC/PDA-QDa) for the qualitative and quantitative analysis of Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and cannabinol (CBN) in confiscated cannabis was developed.The seized cannabis was extracted in methanol by sonication.The binary mobile phase consisted of methanol (containing 0.1% formic acid) and water.After centrifugation, the supernatant was separated on Waters UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) with isocratic elution at a flow rate of 0.2 mL/min.The three cannabinoids were analyzed by photodiode array (PDA) detector at 220 nm and confirmed by mass spectrometer QDa.The correlation coefficient of standard curve for the three cannabinoids in linearity range was not less than 0.999, as well as the recoveries were 82%-102% with the relative standard deviations (RSD) of 0.36%-4.12% at three spiked levels.The method is specific, easy, quick and suitable for confirmation of the cannabinoids in seized cannabis.Cannabis plants in different areas were classified by their chemical phenotype as drug-type or fiber-type plants, taking into account the phenotypic index Δ9-THC, (Δ9-THC+CBN)/CBD, or the Δ9-THC/CBD and the (Δ9-THC+CBN)/CBD ratios.The analysis of the original composition of plant material is necessary for the detection and the quality control of cannabis plants.

AIM:To establish a simple and accurate HPLC method for quality evaluation and determination of 8 active constituents, i.e. Danshensu, Protocatechualdehyde,rosmarinic acid,Salvianolic acid B, Dihydrotanshinone, Cryptotanshinone, Tanshinone Ⅰand Tanshinone ⅡA in Radix Salviae Miltiorrhizae, at the same time, in order to study the correlations of the contents of the 8 constituents. METHODS: Detected at 286 nm, analytes were separated on a ODS C_ 18 column(150 mm?4.6 mm, 10 ?m) eluted gradually with methanol and 1% tetrafuran in 1% formic acid water solution(3∶97)on the increasement of methanol from 3% to 70% in the whole analysis. After determination, SPSS 13.0 software was used to calculate the correlation between the 8 constituents. RESULTS: Under the above conditions, HPLC method was constructed to quantify 8 constituents,and their correlation coefficients were calculated. CONCLUSION: The result shows that the contents of the 8 constituents in Salvia miltiorrhiza Bge. vary greatly from different origins; and there are significant correlations between the contents of hydrophilic constituents such as Salvianolic acid B,Danshensu,rosmarinic acid, so do hydrophobic constituents like Cryptotanshinone,Tanshinone Ⅰ,Tanshinone ⅡA,but there is no correlation between the hydrophilic constituents and hydrophobic constituents.

Objective To investigate the function of amnion endothelial cell in the differentiation of human umbilical cord blood stem cells into dopaminergic neurons.Methods Primary human amnion endothelial cells were separated and cultured in vitro;the conditioned medium(CM) was prepared through high speed centrifugation.The cord blood mesenchymal stem cells of P_1 passage were induced by the conditioned medium,and the mophology of cells was observed under the inverted phase contrast microscope.The expression of tyrosine hydroxylase(TH)and dopamine transportor(DAT) of the induced cord blood mesenchymal stem cells were detected by immunocytochemistry staining method and immunoblotting(Western blotting).Results The masculine rate of TH and DAT of the cord blood mesenchymal stem cells of P_1 passage which were induced by amnion endothelial cells conditioned medium was higher than that of the control group,with a significant difference(P