The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

OBJECTIVETo retrospectivly compare the clinical efficacy of dynamic hip screw (DHS) with proximal femoral nail anti-rotation (PFNA) for the treatment of unstable intertrochanteric fractures in the elderly.

METHODSTotally 92 elderly patients with unstable intertrochanteric fractures were treated with DHS [including 27 males and 23 females with a mean age of (72.5 +/- 5.3) years old] and PFNA [including 22 males and 20 females with a mean age of (72.8 +/- 5.8) years old] from August 2008 to August 2012. The data of operation time,blood loss (obvious and hidden blood loss), bedridden time, down load time, postoperative complications and Harris hip function score were recorded and compared.

RESULTSBoth of two groups were followed-up for 10 to 18 months with an average of 13.5 months. PFNA was implanted with a significantly smaller incision and shorter clinical healing time, less blood loss,while hidden blood loss were more. Postoperative complications, therapeutic effects and Harris score in PFNA group were better than that of DHS group.

CONCLUSIONFor treatment of senile patients with unstable intertrochanteric fractures, PFNA was superior to DHS in reducing complication rates, recovering hip joint, while DHS could reduce perioperative blood loss in treating type II a, II b and III fracture.

Aged ; Aged, 80 and over ; Bone Nails ; Bone Screws ; Case-Control Studies ; Female ; Femur ; surgery ; Fracture Fixation, Internal ; instrumentation ; Hip Fractures ; surgery ; Humans ; Internal Fixators ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective To explore the surgical methods for the treatment of the total anomalous pulmonary venous connection (TAPVC) and choices to prevent related complications.Methods We analyzed retrospectively the clinical data of 24 cases with TAPVC admitted to our hospital from Jan 2006 to Dec 2011,including 15 male and 9 females with the age range of 50 d-14 years.There were 10 cases younger than six months,accounting for 41.7％ (10/24).The average body weight was (9.30 ± 3.96) kg.There were 8 cases 33.3％ (8/24) had a body weight of below 10.00 kg.Among the patients,16 cases (66.7％,16/24) were supracardic type,6 (25.0％) were cardiac type,and 2 (8.30％) were intracardiac type.For the treatment of the upracardiac type,5 cases were treated through the right atrium and interatrial septum incision path;Eleven cases were through the left atrial anastomosis.For the treatment of the cardiac type,the right atrial incision was used for coronary vein antrum isolation,and the patch was carefully packaged to separate the coronary sinus openings into the left atrial side.For the 2 cases of the intracardiac type,heart was slightly lift towards the right,and the venous anastomosis was performed for the left atrial posterior wall and the summary vein,and the vertical vein was then ligated.Results No surgery-related death occurred.Reoperation was performed for 1 patient occurred pulmonary edema due to pulmonary venous obstruction induced two days post-surgery.Condition was improved after the extension of left atrial side as the patient was found to have anastomotic stenosis.Postoperative arrhythmia were observed in 7 cases (29.2％,7/24),including 3 nodal arrhythmia (12.5％,3/24) and 4 (16.7％,4/24) atrial arrhythmia.Patients were followed up for 4-24 months.All children during the follow-up period were in good condition.They had significantly improved activity tolerance compared with pre-surgery.Chest X-ray showed clear markings free of congestion.Conclusion Appropriate surgical approach and routes could help improve the success rate of surgery treatment of TAPVC and reduce postoperative complications,thus achieving good therapeutic effect.

Microbiology is an important,fundamental and obligatory course in contemporary life science.This article introduces that teaching group of microbiology in Nankai University realizes transformation of teaching center,fully embodies the modernization of teaching notion and gives full play to students' main effect practically by adhering to teaching reform as center,optimizing teaching method as measure,communicating in and after class and using multi-media and teaching web.Therefore,teaching system is established to adapt to modern teaching notion and eventually microbiology course becomes a cultivation platform to foster elites with both solid fundamental theory and innovating mind.

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective To investigate the variance of cost-effectiveness when treat acute myocardial infaretion of different severe extents with different pattern.Methods Acute myocardial infarction patients were selected from emergency eommand center of Guangzhou from October 2003 to December 2005.These patients wew assigned by the center to First-Class Hospitals at Grade 3 and First-class Hospital at Grade 2,and were followed up after 6 months after post-discharge.Cost in hospital and mortality in hospital were registered.The health of all patients were quantificated using SF-36.According to the assigned hospitals,the patients were divided into single infarction group and complex infarction group.Cost in hospital,mortality in hospital,short-term quality of life were compared between the them.Results Compared with and First-Class Hospital at Grade 2 (101 cases),the single infarction patients in First-Class Hospitals at Grade 3 had higber costs in hospital (P=0.016),better society function,affection role,mental health and health status (P

Objective To establish an automatic synthesis method for 11C-carfentanil (CFN) as an novel opiate receptor radioligand and study its biodistribution in rats. Methods 11C-Triflate-CH3 was bubbled into 0.5 mg precursor desmethyl-CFN (which was dissolved in 0.15 ml DMSO) to generate 11C-CFN in a V-tube at room temperature. Sep-Pak C2 column was used for purification of 11C-CFN, which was eluted by 3ml binary system aqueous solution, 10 ml water thrice, and then I ml ethanol. The biodistribution (% ID/g) of 11C-CFN in SD rats was studied. SPSS 13.0 was used for statistical analysis. Non-normal distribution data were analyzed using nonparametric test. Results The synthesis time for 11C-CFN was 20 min (end of bombardment, EOB). The synthesis yield was (35.5 ± 2.2) % on average (n = 12, uncorrected)with the radiochemical purity over 98%. Biodistribution study in rats showed that the tracer had a high brain uptake, rapid blood clearance, and a metabolic pathway via liver and kidney. The highest tracer uptake was in thalamus (4.26 ± 0.89) % ID/g and striatum (4.05 ± 1.08) % ID/g at 5 min after injection, followed by cerebral cortex (2.63±0.89) %ID/g, pons (2.26 ±0.57) % ID/g, hippocampus (2. 17 ±0.55) %ID/g and cerebellum (2. 15 ±0.39) %ID/g. Conclusions The automatic synthesis of 11C-CFN is fast and reliable, and this radioligand can be used for opiate receptor imaging.

Pseudorabies virus (PRV), an alpha-herpesvirus, has been used as a vector for live-viral animal vaccines. The recombinant PRV TK- / gE- / GP5+, which expressing GP5 of PRRSV, is developed based on the PRV genetic-depleting vaccine-virus strain, TK- / gE- /LacZ+. However, this strain stimulated poorly the vaccinated animals to produce neutralizing antibodies against PRRSV. In order to develop a booster specific immunized response of the PRV recombinant, the ORF5 gene of PRRSV TK- / gE- / LacZ+ was substituted by a modified ORF5 gene, ORF5m. The resultant recombinant PRV, TK- /gE- / GP5m+, was verified by PCR, Southern blotting and Western blotting. TK- / gE- / GP5m+ and TK- / gE- / GP5+ expressed GP5 proteins were inoculated into balb/c mice to evaluate their immunogenicity. The results demonstrated that the amount of neutralization antibodies and cell-immunity responses induced by TK- / gE- /GP5m+ against PRRSV were higher than that of TK- / gE- / GP5+. This study indicated that the new recombinant PRV expressing the modified GP5m protein is a candidate for the development of bivalent genetic engineering vaccines against PRRSV and PRV.
Animals ; Genetic Vectors ; Herpesvirus 1, Suid ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Porcine respiratory and reproductive syndrome virus ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Viral Envelope Proteins ; biosynthesis ; genetics ; Viral Vaccines ; genetics ; immunology

Objective To investigate the efficacy and tolerability of a recombinant human tumor necrosis factor:Fc fusion protein (rhTNFR:Fc,with a trade name of Yisaipu) in the treatment of moderate to severe psoriasis vulgaris.Methods A multicentre,randomized,double blind,and parallel-controlled trial was performed.One hundred and forty-four patients with moderate to severe psoriasis vulgaris from four centres were randomly assigned and treated with either once-weekly subcutaneous injection of rhTNFR:Fc (50 mg) or oral methotrexate (MTX)(7.5 mg) for 12 weeks.Patients were followed up at 2,4,8,12 weeks after the treatment.Results One hundred and twenty-four patients finished the 12-week course of treat- ment.At 12 weeks after the treatment,a 50%,75%,90% improvement in psoriasis area and severity index (PASI) was achieved by 86.11%,76.39%,52.78% respectively of rhTNFR:Fc-treated patients,and by 63.89%,44.44%,22.22% respectively in MTX-treated patients,and all the three improvement rates were of significant difference between the two groups of patients (all P0.05).Conclusion Compared with MTX,rhTNFR:Fc acts more quickly with a higher cure rate and less toxic reactions in the treatment of psoriasis vulgaris.

This study was purposed to investigate lrp16 gene expression in leukemia cell lines and bone marrow cells of leukemia patients and explore the relationship between lrp16 gene expression and development of leukemia. Reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to test the lrp16 mRNA expression in 4 leukemia cell lines, including K562 (CML), HL-60 (APL), MOLT4 (ALL) and U937 cell lines, as well as in bone marrow-derived cells from 115 patients with leukemia. The effect of lrp16 gene expression on genesis and progression of leukemia was analyzed according to clinicopathological features. The results indicated that positive expression of lrp16 mRNA was found in all 4 leukemia cell lines. For leukemia patients, the positive expression rate of lrp16 mRNA in all AML patients was 38% (16/42), in which the positive rates in AML patients with complete remission (CR) and AML patients without remission were 13% (4/30) and 100% (12/12) respectively. The positive expression rate of lrp16 mRNA in ALL patients was 38% (10/26), in which the positive rate in ALL patients with CR and ALL patients without remission were 16% (3/18) and 87% (7/8) respectively. The positive expression rate of lrp16 mRNA in CML patients was 36% (9/25), in which the positive rates in CML patients with CR and CML patients without remission were 20% (4/20) and 100% (5/5) respectively. The positive rate of lrp16 mRNA in CLL patients was 31% (7/22), in which the positive rate in CLL patients with CR and CLL patients without remission were 11% (2/17) and 100% (5/5) respectively. There was no difference of lrp16 gene expression between leukemia subtypes, but there was statistical significant difference in lrp16 gene expression between CR patients and non CR patients (p < 0.001). It is concluded that the lrp16 gene is a leukemic oncogene and closely relates to genesis and progression of leukemia, which may be an indicator for evaluating clinical efficacy of leukemia therapy.
Adolescent ; Adult ; Aged ; Bone Marrow ; metabolism ; pathology ; Female ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Young Adult