Objective To express recombinant human cardiotrophin 1 (huCT 1) protein with biological activity in E coli Methods huCT 1 open reading frame gene from plasmid pBSSK huCT1 was cloned into plasmid pMD18 T by PCR and T A cloning, and then cloned into prokaryotic GST fusion expression vector pGEX 2T to give pGEX2T huCT1 After pGEX2T huCT1 expression was induced by IPTG in E coli at different temperature and different time, the expression level and the proportion of the soluble protein were analyzed by SDA PAGE Then the soluble GST/huCT1 was purified by immobilized glutathione columns The GST fusion protein was cleaved by thrombin and purified again The recombinant huCT 1 with biological activity was identified according to the survival of axotomized sciatic motoneurons Results After the induction of pGEX2T huCT1 DH5? cells by IPTG at 29 ℃ for 4 h, the highest expression level of the recombinant protein was about 1/5 of total cell proteins, and the soluble portion was about 2/5 of fusion protein Purification of the soluble portion and thrombin cleaved fusion protein resulted in 85% and 80% purified recombinant GST fusion protein and huCT 1 protein, respectively Recombinant huCT 1 protected 55% motoneurons in spinal cord against sciatic axotomy in vivo in adult rats Conclusion Recombinant huCT 1 has biological activity in rat neurons

Objective To construct human cardiotrophin 1(huCT 1) adenovirus vector for central nervous system(CNS) gene therapy in vivo . Methods The huCT 1 and eGFP genes were cloned into shuttle plasmid pDC316 to construct pDC316 huCT 1 and pDC316 eGFP. Virus genome plasmids pBHGloxdeltaE1,3Cre and pHG140 were purified by CsCl banding certification. Recombinant replication defective adenovirus vectors AdCMV huCT1 and AdCMV eGFP were rescued in 293 packaging cells by co transfection and Cre mediated recombination of both plasmids pDC316 huCT1 and pBHGloxdelta1,3Cre. The insert gene and its expression were identified by PCR, RT PCR and immunohistochemistry after recombinant adenovirus transfected 293 cells and NIH 3T3 cells. Recombinant adenovirus vectors were purified by CsCl banding and titrated by plaque forming test. AdCMV huCT1 expression in vivo was analyzed by RT PCR and immunohistochemistry after transfection of the cervical spinal cord in adult rats. Results We have constructed two recombinant adenoviral vectors: AdCMV huCT1 and AdCMV eGFP, containing MCMV promoter, foreign DNA and SV40 PolyA with deletions of E1 and E3 regions. The positive huCT 1 mRNA and protein were identified in AdCMV huCT1 transfected NIH 3T3 cells and rat cervical spinal cord. The titer of virus stocks was generally up to 3.0?10 10 plaque forming units(pfu) per milliliter. Conclusion Recombinant purified AdCMV huCT1 vectors can be highly expressed in vitro and in vivo and is suitable for CNS gene therapy in vivo .

Objective To overcome the disadvantages of the traditional auscultating method and design the electro-auscultating visual system integrating the monitoring and recording functions.Methods The portable system consisted of the module pre-gathering and electro-auscultating heart-sound signal,the circuit of A/D converting and MCU,as well as the device of signal LCD-displaying in real-time and the USB mass storage device.Results Portable electro-auscultating system was realized,which integrated the functions of watching,monitoring and recording.Conclusion The portable system is more convenient for clinical doctors to get correct information from patients,benefiting to the clinical pathology-statistics and profound development of the related data.

OBJECTIVETo explore the effect of microneedle combined with Lauromacrogol on skin capillary network.

METHODS24 male Leghone (1.5-2.0 kg in weight) were randomly divided into three groups as group A (microneedle combined with Lauromacrogol), B (microneedle combined with physiological saline) , and C(control). The cockscombs were treated. The specimens were taken on the 7th, 14th, 21th , and 28th day postoperatively. HE staining, immunohistochemical staining and special staining were performed for study of the number of capillary and collagen I/III , as well as elastic fibers.

RESULTSThe color of cockscombs in group A became lightening after treatment. The number of capillary decreased as showing by HE staining. The collagen I and III in group B was significantly different from that in group A and C (P < 0.05). Special staining showed proliferation of elastic fibers in group B.

CONCLUSIONSIt indicates that microneedle combined with Lauromacrogol could effectively reduce the capillary in cockscomb without any tissue fibrosis. Microneedle can stimulate the proliferation of elastic fiber, so as to improve the skin ageing process.

Animals ; Capillaries ; anatomy & histology ; Chickens ; Comb and Wattles ; blood supply ; drug effects ; Male ; Needles ; Polyethylene Glycols ; pharmacology ; Punctures ; instrumentation ; methods ; Random Allocation ; Skin Aging

Objective To establish a scientific and sensitive evaluation index system of nursing quality for cancer hospital. Methods The evaluation index system of nursing quality for cancer hospital was formulated based on literature,semi- structured interviews and expert group discussion. Then, developing the evaluation index system of nursing quality for cancer hospital by two rounds of Delphi consultation. Results The experts′ authority coefficient was 0.862. The nursing quality indicators included 5 first-level indicators,9 second-level indicators and 73 third-level indicators. The Kendall coordination coefficients of the importance of the three level indicators were 0.354,0.217,and 0.243, The Kendall coordination coefficients of the feasibility of the three level indicators were 0.234,0.313,and 0.339. Conclusions Scientific nature and concentration indicator system of nursing quality for cancer hospital was developed. It will be used to provide quantitative basis for the control of nursing quality in cancer hospital.

Objective To explore the effect of reconstructing unilateral cleft lip by changing the arc-shaped incision, combined with the 3D reconstruction of upper lip muscles.Methods Twenty unilateral cleft lip patients were treated by using a new surgical operation, the 3D reconstruction of upper lip muscle, to restore normal anatomy and stress of the mucous membrane, muscle and skin.Operation scar was designed for straight line, located on the philtral ridges of the contour line;phitrum and philtral ridges were rebuilt, and postoperative scar reduced.Results A lot of 20 patients had no local infection, hemorrhage, complex crack, and were stage I incision healing.Followed up for 1-8 months postoperatively, the patient's lip bow line continuity was good, with symmetrical shape and good phitrum and philtral ridges;scar was hidden on the philtral ridges of the contour line, and no obvious upper lip scar contracture found through the follow-up period.Conclusions This improved method is simple in the incision design, and less scar hidden on the philtral ridges of the contour line after operation, which can maximize the recovery of the appearance of nose and upper lip with satisfactory effect.It is a feasible improvement method of repairing unilateral cleft lip.

It is important to design and build a kinetic loading system for flexing movement of knee joint to study knee biomechanics. The system reported here includes driving device, control device, and flexion angle determination imaging system. The driving device was constructed with a stepper motor and a mechanical transmission with a serried of clamps, shanks and so on, and the driving device was controlled by the control device with micro-control unit, a computer and the serial 232. While the knee joint was driven to move by the stepper motor, the flexion angle of the knee was determined using imaging-based techniques. The system achieved accurate loading and control of speed, extent and duration of knee flexion, as well as fast and non-contract determination of flexion angle during knee flexing movement. The system is simple to build, easy to operate, highly accurate and reliable and it provides an important tool for the study of knee biomechanics, and potentially provides a tool for helping patients of knee surgery during their post operation recovery training.
Biomechanical Phenomena ; Equipment Design ; Humans ; Knee Joint ; physiology ; physiopathology ; Microcomputers ; Orthotic Devices ; Range of Motion, Articular ; physiology ; Stress, Mechanical ; Weight-Bearing ; physiology

OBJECTIVETo investigate the cytogenetic differences between children and adults with acute lymphoblastic leukemia (ALL) using eight-probe fluorescence in situ hybridization and karyotype analysis.

METHODSEight-probe (MYC, P16, E2A, TEL/AML1, BCR/ABL , MLL , IGH, and hyperdiploidy) fluorescence in situ hybridization and karyotype analysis were performed for 86 adults and 39 children with acute lymphoblastic leukemia.

RESULTSEight-probe fluorescence in situ hybridization showed significant differences in the positivity rate of TEL/AML1, BCR/ABL, and hyperdiploidy between adult patients and children with ALL. By karyotype analysis, the positivity rate of t(9;22) and hyperdiploidy differed significantly between the children and adult patients (P<0.05).

CONCLUSIONAdults and children with ALL have different expression profiles of the fusion genes. Eight-probe fluorescence in situ hybridization is time-saving, accurate and efficient in detecting common genetic abnormalities in ALL patients, and can be well complementary to karyotype analysis in clinical diagnosis of ALL.

Adolescent ; Adult ; Child ; Child, Preschool ; Cytogenetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Infant ; Karyotype ; Karyotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics ; Young Adult