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.

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.

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.

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 investigate the effects of bone marrow stem cells autologous transplantation after acute myocardial infarction on local mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1and VCAM-1.Methods Acute myocardial infarction models were established in 10 swine,which were randomized divided into placebo group (5 swine) and transplantation group (5 swine).After six months,hearts of placebo group and early transplantation group were harvested and mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1 and VCAM-1 in infarcted myocardial region was detected.Results The mRNAs of SDF-1,Kit ligand,E-selection,ICAM-1 and VCAM-1 were expressed in the infarcted myocardial region of transplantation group six months after myocardial infarction,but it was not found in the placebo group.ConclusionsBone marrow autologous transplantation after acute myocardial infarction can promote local mRNA expression of SDF-1,Kit ligand,E-selectin,ICAM-1 and VCAM-1,which may be due to the paracrine effects of stem cells.

Objective To evaluate the current situation of two-way referral system by qualitative study.Methods The qualitative study was carried out in Department of General Internal Medicine of Peking Union Hospital and 4 clinics under Beijing Gaobeidian Community Health Center from June to December 2009.In-depth interview was conducted on 6 patients,who were referring from community health center to tertiary hospital and 16 general practitioners in the community.The method of thematic framework was used for data analysis.Results Only 1 of the 6 patients,who made the appointment,completed the two-way referral.From the view of the patients,the interrupt of the two-way referral was due to long-time waiting,inconvenient transportation to the hospital,availability of specialists,unsmooth process of referral and unsatisfied doctors.However,the community health service staff thought that difficulties of the two-way referral originated from the system,doctors,patients,and doctor-patient relationship.Sometimes patients distrusted doctors both in hospital and in community; communications between doctors or between doctors and patients were less efficient; patients had multiple choice for their primary care and they may go to any hospital freely bypass community-based referral system; patients had excessive expectation for medicine and doctors; some patients paid less attention to their own health,and so on.For improvement of the two-way referral system,the community health service staff proposed concrete measures in terms of relevant policy,the hospital,community health service,the doctors and patients; they emphasized the importance of upgrading their own communication ability and professional skills for primary care.Conclusions Many factors may influence successful implement of two-way referral,so it is necessary to improve the relevant policy and procedures and to upgrade medical staff in community.