To rapidly and accurately manipulate genome such as gene deletion, insertion and site mutation, the whole genome of a very virulent strain Md5 of Marek's disease virus (MDV) was inserted into bacterial artificial chromosome (BAC) through homogeneous recombination. The recombinant DNA was electroporated into DH10B competent cells and identified by PCR and restriction fragment length polymorphism analysis. An infectious clone of Md5BAC was obtained following transfection into chicken embryo fibroblast (CEF) cells. Furthermore, a lorf10 deletion mutant was constructed by two step Red-mediated homologous recombination. To confirm the specific role of gene deletion, the lorf10 was reinserted into the original site of MDV genome to make a revertant strain. All the constructs were rescued by transfection into CEF cells, respectively. The successful packaging of recombinant viruses was confirmed by indirect immunofluorescence assay. The results of growth kinetics assay and plaques area measurement showed that the lorf10 is dispensable for MDV propagation in vitro. Overall, this study successfully constructed an infectious BAC clone of MDV and demonstrated its application in genome manipulation; the knowledge gained from our study could be further applied to other hepesviruses.
Animals ; Chick Embryo ; Chickens ; Chromosomes, Artificial, Bacterial ; DNA, Recombinant ; Herpesvirus 2, Gallid/genetics* ; Marek Disease

Objective To establish the three-dimensional (3D) finite element model of unicompartmental knee arthroplasty (UKA) with 3° and 7° posterior tibial slope at different knee flexion angles, and to study biomechanical properties and prosthetic wear of the knee joints with two types of posterior tibia slope and their effects on knee function. Methods Combining CT and MRI images of human knee joints with the 3rd-generation Oxford prosthesis, the finite element UKA model with 3° and 7° posterior tibia slope were established. The 1 kN load was applied to center point of the medial and lateral condyles of the femur to simulate the standing load of human body. The maximum stresses and distributions of the prosthesis and articular cartilage at different knee flexion angles were analyzed. ResultsThe maximum stress of the meniscus liner with 3° posterior tibia slope at 0°, 30°, 60°, 90°, 120° knee flexion angles increased by 28.06%, 68.99%, 19.45%, 21.06% and 53.38%, the distribution area was concentrated from the side of the meniscus liner to the central area, and the stress concentration was obvious at 120° knee flexion. The maximum stress of prosthesis with 3° posterior tibia slope was greater than that with 7 ° posterior tibia slope. The expansion of stress concentration area would cause wear and loosening of the prosthesis, contact stress and concentration area of the articular cartilage would subsequently increase with posterior tibia slope increasing, and stress concentration would be more obvious at high knee flexion angles. Conclusions Tibial prosthesis has the higher stress and greater wear under the condition of 3° posterior tibia slope than 7° posterior tibia slope. The research findings provide theoretical basis for the UKA design in clinic.

Objective To apply the Chinese version of Care Partner-Frailty Index-Comprehensive Geriatric Assessment (CP-FI-CGA) in Taiyuan part of elderly patients and study theirs frailty conditions. Methods To Choose the Chinese version of CP-FI-CGA questionnaire, patients′ general information questionnaire and Clinical Frailty Scale to evaluate 385 patients and analyze the results statistically. Results Of 385 patients, female patients were 166 cases (43.12%);the frailty index score was (0.318 ± 0.165) points, the CFS was 5.044 ± 1.483. Single factor analysis showed that age, marital status, the kinds of medication which the patients used, how much help the patient required, the condition of social support, and the sleep state, these six factors had statistical significance (Z=-7.292, Z=-1.994, χ2=27.726, Z=-9.688,χ2=8.117,χ2=53.477, all P<0.01). Multiple linear regression analysis showed that age, the kinds of medication which the patients used, how much help the patient required and the sleep state, these four factors were independent factors (model R=0.610, R2=0.372; adjusted R2=0.362, F=37.241, P< 0.01). Conclusions CP-FI-CGA questionnaire can accurately estimate the frailty degree by evaluating patients′each system and can be promoted in the clinical geriatric ward and nursing home, etc.

BACKGROUND:Intramedul ary nails and locking plates are widely used for two-part proximal humerus fracture. Which is better for two types of implants in patients remains controversial. OBJECTIVE:To determine the clinical outcome of intramedul ary nails versus locking plates for two-part proximal humerus fracture according to Cochrane Meta analysis. METHODS:We searched PubMed, SCI, Embase, the Cochranel Library and CBMdisc, VIP information, Wanfang Database, and CNKI for randomized control ed trials and quasi-randomized control ed trials on intramedul ary nails and locking plates for two-part proximal humerus fracture. RevMan 5.2 software was used to analyze operation time, intraoperative blood loss, fracture healing time, postoperative complications (heterotopic ossification, pain, screw penetration, necrosis of humeral head), and Constant Score. RESULTS AND CONCLUSION:Six articles of clinical control ed trials were included with 259 patients. 131 patients received intramedul ary nails, and 128 patients received locking plates. Meta-analysis displayed that no significant difference in fracture healing time, heterotopic ossification, pain, necrosis of humeral head and Constant Score was detected between intramedul ary nails and locking plates for two-part proximal humerus fracture. Operation time, intraoperative blood loss, and screw penetration were significantly less in the fixation with intramedul ary nails than that in locking plates (P<0.05). These findings suggested that compared with locking plates, intramedul ary nails method for two-part proximal humerus fractures could reduce screw penetration.

Objective:To prepare and identify the mouse anti-human monoclonal antibodies ( mAbs) using leukocytes as im-munogens. Methods: The mice were immunized using human peripheral blood leukocytes. Then, use of B lymphocyte hybridoma technology preparation of mAbs,followed screening by immunocytochemistry and limited dilution. The secreted mAbs were identified by immunoprecipitation,mass spectrometry,Western blot,ELISA and immunohistochemistry. Results:The 35 positive polyclonal cells were obtained,of which 11 strains secreted mAbs against S100A9. And one strain was used to prepare monoclonal antibody. The purified mAb against S100A9 were purified and identified as IgG1 subtype,with the titer,purity and affinity constant was 1∶3. 18×105,95% and 3. 54×108 L/mol,respectively. This mAb generally had 0. 12% crossed reactivity to S100A8 ,and showed little or no cross reactivity to S100A12 and S100A13. The prepared monoclonal antibodies can specifically recognizes the S100A9 antigen in human breast cancer tissues. Conclusion:Successful preparation of mAb against S100A9,which can secrete specific mAb against S100A9 protein with high titers and specificity have been established successfully,which laid the foundation for the immunology application.

BACKGROUND:Some control studies attempt to answer the advantages and disadvantages of high-dose methylprednisolone sodium succinate therapy for acute spinal cord injury in adults, but have arrived at different conclusions. OBJECTIVE:To explore the therapeutic efficacy of high-dose methylprednisolone sodium succinate therapy on acute spinal cord injury in adults by Meta analysis. METHODS:PubMed, Embase, Cochranel Library, CBMdisc, VIP and WanFang Databases were searched by computer, and relevant Chinese and English orthopedic journals were retrieved by hand. Controled trials related to high-dose methylprednisolone sodium succinate therapy of acute spinal cord injury in adults were included. The methodology quality of included trials was criticaly assessed. RevMan 5.0 software was used for data analysis. RESULTS AND CONCLUSION:Nine clinical controled trials were included. Meta-analysis results showed that compared with the conventional therapy, the neurological recovery rate after 24 hours of administration, pneumonia incidence and gastrointestinal reactions increased significantly after high-dose methylprednisolone sodium succinate therapy. However, there were no statistical differences in the rate of urinary tract infection, nonunion rate and stress ulcer incidence between these two therapies. These findings indicate that the high-dose methylprednisolone sodium succinate therapy on acute spinal cord injury in adults has better outcomes in neurological function recovery, but can lead to higher incidence of lung infection and gastrointestinal reactions. Therefore, lung infection and gastrointestinal reactions should be avoided as much as possible during the course of treatment.

Objective To determine the prevalence of diabetes mellitus and related risk factors among residents in the urban and rural area of Chengdu.Methods A cluster sampling was used to establish a study population of inhabitants aged 30 to 70.Totally,1 847 participants were enrolled in this study.Questionnaire including general information and dietary information in the past year was used to collect related data.Height,body weight and oral glucose tolerance test (OGTT) were measured.Factor analysis was used to analyze the dietary pattern while multivariate unconditional logistic regression used for risk factors in total population,urban and rural residents,respectively.Results The population standardized prevalence rates of diabetes in the overall,urban and rural residents were 20.2％,28.7％,11.1％,respectively.Among total population,middle-aged (OR=2.337,95％CI:1.305-4.185) and the elderly (OR=5.990,95％ CI:3.389-10.586) residents had higher diabetes risk than the younger ones.Administrators (OR=1.434,95％ CI:1.000-2.057) and ordinary clerks (OR=2.870,95％ CI:1.653-4.980) were more vulnerable to diabetes than peasants.Similarly,middle-aged (OR=2.973,95％ CI:1.101-8.031) and elderly (OR=5.972,95％ CI:2.267-15.730) turned out to be more predisposed than young people in the urban area.Compared with peasants,ordinary clerks (OR=2.196,95％CI:1.213-3.975) seemed to be more liable to diabetes.In the mral areas,dietary pattern with higher energy and protein (OR=1.404,95％ CI:1.113-1.772) could be subject to diabetes.Conclusion The prevalence of diabetes in Chengdu was relatively high.Age,career and dietary pattern are mainly risk factors.The factors in different districts are vaious.Intervention on nutrition should be different,area-wise.

BACKGROUND:Intramedul ary nail and traditional steel plate fixation have been widely used in the treatment of distal tibial (extra-articular) fractures, but these two kinds of methods have shortcomings. Some control studies attempt to answer the advantages and disadvantages of the issue on the treatment of extra-articular distal tibial fractures with intramedul ary nail and steel plate fixation, but the conclusions are different.
OBJECTIVE:To determine the efficiency and safety of intramedul ary nail and plate fixation in the treatment of extra-articular distal tibial fractures by Meta analysis.
METHODS:PubMed, Embase, Cochrane Library, CBMdisc, Weipu information database and Wanfang were searched by computer, and relevant Chinese and English orthopedic journals were hand-searched. Control ed trials related to intramedul ary nail and plate fixation in the treatment of extra-articular distal tibial fracture were included. The quality of trials was critical y assessed. RevMan 5.0 software was used for data analysis.
RESULTS AND CONCLUSION:Four articles were included. Meta analysis showed that compared with the plate fixation, the malunion rate of the intramedul ary nail for extra-articular distal tibial fractures increased. There was no statistical difference in the rate of nonunion rate, delayed fracture healing, and infection rate after operation. These indicate that intramedul ary nail and plate fixation both have good effects on fracture healing.

Background and objective Th17 cells are important T helper cells, which are characterized by their production of IL-17. hT17 cells play an important role in host defense against microbial infections, autoimmune diseases and cancer. hTe aim of this study is to investigate the percentage of hT17 in peripheral blood lymphocyte and the level of IL-17 in serum and cerebrospinal lfuid (CSF) in patients with brain metastases from lung cancer. Methods Twenty-two patients with brain metastases from lung cancer and 20 health controls were analyzed. hTe percentage of hT17 cell was detected with lfow cytometry using CD3+CD4+IL-23R+marker, the level of IL-17 was measured with ELISA method. Results hTe percentage of hT17 cells in patients with brain metastases from lung cancer was 4.65%±0.72%, which was remarkably higher than that in con-trols (2.71%±0.54%, P=0.04). hTere was no signiifcant difference between non-small cell lung cancer (NSCLC) patients and small cell lung cancer (SCLC) patients. Serum IL-17 was remarkably increased in patients with brain metastases from lung can-cer (117.4±16.43 pg/mL vs 72.55±8.19 pg/mL, P=0.02). No signiifcant difference of the serum IL-17 was observed between NSCLC and SCLC patients. hTe level of IL-17 in CSF from patients with brain metastases from lung cancer was signiifcant higher than that from lung cancer patients without brain metastases (73.21±7.52 pg/mL vs 50.25±8.04 pg/mL, P=0.04). Con-clusion hT17 cells and IL-17 increase in patients with brain metastases from lung cancer. It may involve in the pathogenesis of brain metastases from lung cancer.

Objective To explore the CT findings, the clinicopathological features of giant lymph node hyperlasia and its combined paraneoplastic pemphigus. Methods The clinical features,the imaging and the follow-up data of 19 patients surgically confirmed as giant lymph node hyperlasia were analyzed. Results Clinically, the majority of patients not accompanied with paraneoplastic pemphigus abnormal pulmonary,showed asymptomatic, and only a very small number had lower abdominal discomfort or pain.The patients associated with paraneoplastic pemphigus and abnormal pulmonary manifested the special clinical manifestations, CT finding, pathological characteristics. Histopathologically, the hyaline-vascular type was found in 18 cases and mixed type in 1 case. CT scanning showed that the lesions in 18 patients appeared as larger(2.5-15 cm in diameter), solitary, cylindrical soft-tissue masses with marked enhancement. One case was presented as multiple enlarge lymph node at left neck (1.5-5 cm in diameter). Seventeen of 19 cases were smooth at the edge, and ten cases were uniform in density. The calcification was characterized of an arborizing and (or) flocculent pattern and central location in 7 cases, of which, 1 case circumferential distribution and 5 cases scattered with multiple spots or strip. Shapes were cylindrical and spherical or elliptical. All patients with giant lymph node hyperlasia showed marked enhancement after contrast administration at arterial phase and delay scan. Conclusion CT scanning is an effective method in diagnosis, guiding surgery and evaluating prognosis of giant lymph node hyperlasia, especially dynamic contrast-enhanced and delayed CT scanning. It is the critical way for patients complicated with paraneoplastic pemphigus and abnormal pulmonary cases, to diagnose early and resect tumor-like lesions in vivo.