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.

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.

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.

ObjectiveTo evaluate the security and effectiveness of minimal invasive fixation with 3D fluoro-images navigation in the management of acetabular fractures.MethodsFrom August 2008 to December 2010,20 patients with acetabular fractures were treated with percutaneous screw fixation under the guidance of 3D fluoro-images based on navigation system after closed reduction.There were 9 males and 11 females,aged 26-54 years old(mean,37.1±1.2 years).Fractures were caused by traffic accident in 15cases,and falling from height in 5 cases.According to AO classification,there were 2 cases of A1.1 type,3 cases of A2.2 type,4 cases of A2.3 type,2 cases of A3.2 type,2 cases of A3.3 type,2 cases of B1.1 type,2 cases of B2.2 type,1 case of B3.1 type,1 case of C2.1 type,1 case of C2.3 type.The interval from injury to hospitalization was 2-46 h (mean,8.6±0.2 h).After 3-14 days of skeletal traction through tibial tubercle,the operation was performed.ResultsTotally 46 screws were fixed.The average time for surgery was(22.6 ±1.2) min per screw.Forty-five screws were placed correctly with a successful rate of 97.8％,only one screw was reinserted for deviation.All the screws were checked by the 3D fluoro-images that they were not in the joint space after fixation.The result was same with CT scan.No incision problem and implant failure occurred.All 20 patients were followed up 12 to 28 months with an average of (22.8±1.7) months.At last follow-up,fracture union was achieved in all patients with satisfactory screw fixation.ConclusionThe minimal invasive fixation with 3D fluoro-images navigation makes the surgery for the nondisplaced acetabular fracture more precise and time-saving,minimize the surgery injury,and improves clinical results without an increasing rate of complications.

Objective To evaluate the effect and potential risks of two types of materials ( taitanum versus polylevolactic acid,PLLA) screws on fixation of distal tibiofibular syndesmosis by a prospective randomized comparative clinical study.Methods From January 2007 to May 2008,48 patients with ankle fractures combined with a verified syndesmotic rupture were randomly allocated to receive either bioabsorbable PLLA (n =28 ) or titanium (n =20) screw fixation.The patients were assessed in aspects of radiography,function recovery and complication during follow-up.Results The total followup rate was 98％.The demographic information of the two groups showed no statistical difference ( P ＞0.05).After 1-2 years of follow-up,the indexes including Baird score,total score,and Baird grade were not statistically different between the two groups ( P ＞ 0.05).At the end of follow-up,there was significant difference in regard of joint motion between the injured and uninjured ankle in both groups ( P ＜0.05).Moderate rejection occurred in two patients and tibiofibular heterotopic ossification in two in PLLA group.Conclusions There is no significant difference between the bioabsorbable and metallic screws with regard to ankle function recovery,but the bioabsorbable screw fixation can avoid a second operation for taking out the screw.However,the complication risk of the bioabsorbable screws should not be ignored,such as rejection reaction,heterotopic ossification.

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.

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.