In order to improve the expression efficiency of recombinant IL~6 in eukaryotic cells, we constructed a new vector pcDIIL-6 to express IL-6 in eukaryotic cells on the basis of an eukaryotic vector pcDNA3IL-6 by introducin g the first intron of human?-globin gene into the downstream of CMV promoter in the pcDNASIL - 6. The IL-6 level expressed by pcDIIL - 6 is 7. 5 times higher than that of pcDNA3IL - 6. On the other hand , we studied the influence of pAdVAntage vector co - transfection on the expression level of IL-6 in eukaryotic cells and found the level of IL-6 is 3. 6 times higher than that of pcDNA3IL - 6. The results we got are beneficial to express cytokines with high efficiency in eukaryotic cells.

Objective To investigate the effects of extracorporeal membrane oxygenation (ECMO) in severe cardiopaths with crisis. Methods Fourteen cases of severe eardiopaths with crisis were treated by ECMO with V A- ECMO technique, whose pump was centrifugal pump and whose tubes was spread byheparin. The cases included 8 fulminating myocarditis (FM) cases with ventricular arrhythmias or/and acute heart failure or/and cardiac shock and 6 acute myocardial infarction cases with pump failure or cardiac shock,in whom 10 cases with cardiorespiratory resuscitation. Haemodynamics and blood gas analysis, and so on were measured before and after treatment. Results The support time of ECMO was 3-106 h, mean (32.4±27.6) h. After ECMO mean arterial blood pressure (MAP), left ventricular ejection fraction (LVEF), left ventricular end-diastolic dimension (LVEDd), partial pressure of arterial oxygen, saturation of blood oxygen improved (P < 0.05 or < 0.01 ), negative value of base excess decreased significantly( P < 0.01 ). MAP had notchanged before and after stopping ECMO [ (80.02±10.20) wan Hg (1 mm Hg = 0.133 kPa) vs (76.34±9.15) mm Hg] (P > 0.05), however, LVEF and LVEDd improved continually (P <0.05). Conclusion ECMO can provide oxygen supply and stable circulation volume for severe cardiopaths with crisis to recover cardiorespiratory function or save valuable time to treat primary disease.

Contents in orthopaedic are independent relatively and finding out the internal relations during them is helpful to improving the effect of orthopaedic teaching.Transfer theory is important tools for clinical teaching practice,and finding out the common characteristics between the orthopaedic chapters is primary for the theory.This research focuses on the following fields:fractures,nerve injury,infection,tumor and deformity.After the common characteristics between these chapters were analyzed and discussed,we concluded that the transfer theory is helpful in orthopaedic teaching practice,especially for students' comprehension and memory,but still we should avoid some negative effects in teaching process.

The evaluation systems for chronic venous insufficiency of lower extremities play a very important role in standardizing the diagnosis and evaluating the curative effect in patients with the disease. This paper gives an introduction to the CEAP classification system, VSS evaluation system and chronic venous insufficiency questionnaire.

Phytoene synthase is a rate-limiting enzyme in the pathway of carotenoid biosynthesis.To aim at establishing a transformation of Ginseng callus cells, elevating the nutritive value through encouraging the composition of corresponding carotenoid,taking Ginseng callus cells as acceptor,psy gene were transformed into cells via Agrobacterium-mediated. The factors affecting genetict ransformation were also investigated seperately from concentrations of A.tumefaciens, age of Ginseng callus cells, infection time and co-culture time. PCR,PCR-Southern and RT-PCR analysis from the transferred gene plants proved that psy gene has been transferred into Ginseng callus cells and can be expressed. The content of ?-carotene was increased by an average of 26 times.A base of improving and raising the contents of carotenoid in the Ginseng callus cell was established.

Objective To establish a real-time fluorescence quantitative methylation assay to investigate the methylation status of GSH-sulphur-transferase P1(GSTP1) gene promoter region in hepatocellular carcinoma(HCC) and to investigate whether which can be used as the early diagnostic indicator of HCC .Methods Ninety-five serum samples were collected from 40 patients with HCC ,30 patients with liver cirrhosis and 25 individuals with healthy physical examination as controls .The methylation level of GSTP1 gene in these serum samples were quantitatively determined by using the real-time fluorescence quantitative methylated spe-cific PCR technique .The receiver-operation characteristic(ROC) curves were adopted to evaluate its diagnostic value for HCC .Re-sults The methylation quantitative level of GSTP1 gene in HCC serum was significantly higher than that in the healthy controls (P<0 .05) .The ROC curve analysis demonstrated that the methylation quantitative analysis of GSTP1 gene could efficiently distin-guish HCC and cirrhosis from healthy controls (AUC=0 .8641) .With the methylation rate of 2% as the critical value for diagno-sing HCC ,its diagnostic specificity was 87 .5% ,the sensitivity was 69 .6% ;the combination detection of serum GSTP1 gene methy-lation and serum AFP could increase the detection rate of HCC to 75% .Conclusion The real-time fluorescence quantitative methyl-ation assay can accurately quantify the methylation level of serum GSTP1 gene ,which has certain application value for the early di-agnosis of HCC .

Objective To evaluate the effect of type Ⅱ odontoid fracture,type Ⅰ Hangman fracture,C2-C3 disc injury on stability of C1-C2 and C2-C3 segments and investigate the clinical significance.Methods Ten fresh-frozen cadaveric cervical specimens (5 men and 5 women; 25-45 years of age,mean 35.7 years) were selected to test the stability of C1-C2 and C2-C3 segments in the settings of intact condition (control group),type Ⅰ Hangman fracture,type Ⅱ odontoid fracture,type Ⅰ Hangman fracture + type Ⅱ odontoid fracture,type Ⅰ Hangman fracture + type Ⅱ odontoid fracture + C2-C3 disc injury.Range of motion (ROM) and neutral zone (NZ) of those segments were measured.Results Compared with the intact condition,type Ⅰ Hangman fracture produced no significant variations in C1-C2 ROM in all loading modes and C2-C3 ROM during left and right lateral bending; type Ⅱ odontoid fracture produced no significant variations in C2-C3 ROM in all loading modes and C1-C2 ROM during left and right rotation; type Ⅰ Hangman fracture + type Ⅱ odontoid fracture revealed no significant variations in C1-C2 ROM during left and right rotation and C2-C3 ROM during extention; type Ⅰ Hangman fracture + type Ⅱ odontoid fracture + C2-C3 disc injury produced no significant variations in C1-C2 ROM during left and right lateral bending and extension-flextion and C2-C3 ROM in all loading modes (P ＜ 0.05).Conclusions Type Ⅰ Hangman fracture can reserve C1-C2 segmental left and right rotation and extension-flextion; type Ⅱ odontoid fracture can reserve C1-C2 segmental left and right lateral bending and extension-flextion; type Ⅰ Hangman fracture + type Ⅱ odontoid fracture + C2-C3 disc injury can reverse atlantoaxial rotationary stability and C1-C3 segmental stability in all directions.This study provides the biomechanical basis for clinical treatments and the related researches of internal fixation.

Objective To investigate the stress characteristics of atlanto-axial bony structure under conditions of anteflexion,posterior extension,lateral flexion,and rotation after artificial atlanto-odontoid joint arthroplasty using three-dimensional finite element method and to improve the orientation of artificial atlantoodontoid joint from perspective of stress.Methods A three-dimensional finite element model of prosthetic atlanto-odontoid joint arthroplasty was created from CT images of the artificial atlantoodontoid joint and cervical vertebrae using software Mimics,Freeform,and Ansys.Stress characteristics of the model dealt with proneness,posterior extension,lateral flexion,or rotation loads were observed.Biomechanical performance of the bony structure of the model was analyzed and the orientation in improving the prosthesis was discussed.Results Anteflexion loading produced a maximum stress of 0.138 ×l08 N/m2 at the junction of lateral mass and posterior arch of the atlas,and 0.201 × 108 N/m2 at axial nail hole,contact point of plates with the axis,and posterior arch of the axis.Posterior extension loading produced a maximum stress of 0.666 × 107 N/m2 at junction of lateral mass and posterior arch of the atlas and 0.254 × 108 N/m2 at arch of the axis.Besides,stress concentration occurred at atlantoaxis nail hole.Right bending produced a maximum stress of 0.124 × 108 N/m2 at nail hole of right mass of atlas and 0.178 × 108 N/m2 at right contact point of the axis with plates.Right rotation produced a maximum stress of 0.847 × 107 N/m2 at junction of lateral mass and posterior arch of the atlas and 0.170 × 109 N/m2 at contact point of the axis with plates.The finite element model comprised 28 620 nodes and 107 441 units and provided good defining of the structural properties of artificial atlanto-odontoid joint arthroplasty.Under different loading conditions,the stress was mainly distributed in contact point of the vertebral body with plates,nail holes,junction of lateral mass and posterior arch of the atlas,and axial pedicle.Conclusions Prosthetic atlanto-odontoid joint scatters a part of the stress and alters the stress distribution of the atlas and axis from the intact condition.Finite element method can obtain complete analysis of the stress distribution of the artificial atlanto-odontoid joint arthroplasty.

AIM:To select an optiomal formulation of liposome entrapped alkanin prepared with ethanol injection in the combination of orthogonal design and central composite design/Response surface method.METHODS:Five influencing factors,including ratio of alkanin to liposome and soyabean lecithin to cholesterol,volume ratio of ethanol phase to water phase,soybean concentration and mannitol concentration were chosen as indices by orthogonal design.Then the method utilized the central composite design,which fitted multivariate linear and second/third order nonlinear regression equation,to find out the optimal formulation.RESULTS:Third order regression equation gained made the good prediction about entrapment(85.4%)and alkanin load(5.16%),and exhibited no significant difference between theoretic value and measured value.CONCLUSION:The combination of orthogonal design with central composite design is a feasible and convenient method to optimize the prescription of alkanin-liposome.

BACKGROUND: Primate is the best animal to establish a model of spinal implants. However, ethics and cost limit its application. Mixed-breed dogs have similar anatomic structures as human. Moreover, it is easy to obtain with low cost, so it may replace primate to serve as models.OBJECTIVE: To determine the feasibility of canine lumbar spine to establish the anterior interfixation model following measurements of related anatomic data.METHODS: A total of 9 adult healthy dogs were selected. The transversal diameter, sagittal diameter and height of vertebral body and disc were measured, and the artificial vertebral body replacement was performed. Bone graft fusion was observed. RESULTS AND CONCLUSION: The transversal diameter, sagittal diameter and height of vertebral body and disc increasedgradually from L_(1) to L_(7), and the height was greatly exceeded its sagittal diameter. All dogs survived, but one was paraplegia. Allthe others stood and acted in 12 h to 72 h after operation. The fusion effect was proved to be good by imageology and histology. The establishment of models was simple and cost-effective, and the biocompatibility of bone tissues and implants, as well as thebone tissue ingrowth can be observed. The lumbar spine of dog can be used as an anterior interfixation model in vitro test.