AIM: DNA vaccines expressing protective domain of surface protective antigen A（spaA）of Erysipelothrix rhusiopathiae have been relatively ineffective at generating high-titer, long-lasting, neutralizing antibodies in murine models. METHODS: This paper report using a DNA vaccine expressing a fusion of the spaA protein and various elements, such as a secretion leader sequence from the highly expressed human gene encoding α1-antitrypsin (AAT), a highly soluble and stably folded domain from the rat cartilage oligomerization matrix protein (COMP), and three copies of the complement component, C3d3, to enhance the titers of neutralizing spaＡ-specific antibody. RESULTS: Analysis of titers of the antibody raised in vaccinated mice at different time points indicated that immunizations with the DNA expressing pcDNA3-AAT-COMP-spaAN-3C3d((pcD-ACSC)) had higher titers than pcDNA3-spaA_N(pcD-S) at weeks 4. Furthermore, the immune protective efficacy of the spaA-chimeras was demonstrated by lethal challenge with a virulent homologous strain 1249 against immunized mice. CONCLUSION: These results suggest that using a plasmid vector containing a strong heterologous signal sequence that mediate efficient antigen secretion in vivo and a fused piece of sequence improving antigens solubility, as well as C3d3, genetic adjuvant, could enhance the antibody responses level. This approach might be an efficient way to improve the antibody level of spaA_N DNA vaccination.

Objective To observe the effect of cinobufotalin freeze-dry powder on heart rate ( HR ) and electrocardiogram ( ECG) of SD rats and to provide experimental basis for monitoring its adverse effect on heart in clinical application. Methods The drug was administered into external jugular vein at constant speed throughout the whole experiment;standard-Ⅱ limb lead monitored the HR and ECG, and then the changes in HR and ECG before and after administration of cinobufotalin were compared. Results Thirty minutes after administration of cinobufotalin injection and cinobufotalin freeze-dry powder at middle dose and high dose, HR of the rats was significantly increased as compared with blank control group[(469±40) bpm, (466±29) bpm and (484±40) bpm vs. (411±17) bpm] (P<0. 05), but soon afterwards, it returned to normal. A small number of rats also developed arrhythmia. In addition, administration of cinobufotalin significantly shortened the P-R interval[blank control group: (46. 90±3. 90) ms, cinobufotalin injection administration group: (39. 70± 2. 54) ms;middle dose of cnobufotalin freeze-dry powder administration group: (37. 70±3. 77) ms; high dose of cinobufotalin freeze-dry powder administration group:(39. 30±7. 12) ms] and Q-T interval[blank control group:(61. 29±9. 46) ms;middle dose of cinobufotalin freeze-dry powder administration group: (55. 13±4. 67) ms; high dose of cinobufotalin freeze-dry powder administration group:(51.75±11. 53) ms] (P<0. 05), but QRS and S-T were unchanged (P>0. 05). Conclusion Cinobufotalin freeze-dry powder has some side effects on rat heart and can increase HR, even lead to arrhythmia.

Objective To summarize complications and early clinical effect of less invasive stabilization system and the femoral condylar support plates in treatment of AO type C distal femoral fractures. Methods We reviewed 46 patients who had internal fixation of AO type C distal femoral fractures. Of all 46 patients, 25 were with less invasive stabilization sys-tem and 21 were with femoral condylar support plates fixation. Comparative analysis was performed using intraoperative in-dex, postoperative complications and the Evanich score at follow-up. Results All 46 patients were followed up with a mean time of 19.6 months after surgery. The difference in incision length, blood loss, fracture healing time was significant between the 2 groups(P＜0.05)but not in the duration of operations and hospital stays(P>0.05). The statistical signifi-cance was also found in the total incidence of postoperative complications and the Evanich score at the last follow-up(P＜0.05). Conclusion Patients with less invasive stabilization system fixation had the characteristics of less trauma, shorter fracture healing time, less postoperative complications and better functional recovery compared with femoral condylar sup-port plates. Less invasive stabilization system had became an ideal internal fixation in treatment of AO type C distal femoral fractures.

Objective To investigate the related mechanism of ligamentum flavum (LF) hypertrophy in diabetic pa-tients with lumbar spinal canal stenosis ( LSCS ) .Methods Twenty-four diabetes mellitus patients [ DM (+) ] and twenty normoglycemic patients [ DM (-) ] with LSCS were enrolled in this study .Sorbitol in LF was analyzed using D-Sorbitol/Xylitol test kit .The thickness of LF was measured by CT .The structure of LF was observed after HE and Masson's trichrome staining .The cell cycle and proliferation of fibroblastic cell NIH 3T3 line cultured in high glucose were analyzed .Sorbitol of NIH3T3 was detected under different backgrounds in vitro, normal glucose , high glucose and high glucose burdened with aldose reductase inhibitor ( ARI) , Epalrestat .The expression of inflammatory factors was detected by qPCR and Western blot under above different backgrounds .Results LF of diabetic patients exhibi-ted significantly higher level of sorbitol and pro-inflammatory cytokines , TGF-βand of CD68-positive staining than that of the normoglycemic subjects ( P<0.01 ) .The diabetic LF was significantly thicker than that of the controls , and showed evidence of degeneration .The high glucose-cultured fibroblasts exhibited significantly higher levels of sorbitol , pro-inflammatory factors , and TGF-βcompared to the low glucose-cultured cells , and these levels were dose-dependently reduced by treatment with the aldose reductase inhibitor (P<0.05).Conclusions Sorbitol level of the LF is significantly increased in the DM patients with LSCS .Increased sorbitol recruites inflammatory factors and fibrogenic-related factor TGF-βin LF of DM patients with LSCS which may contributes to the LF hypertrophy .

BACKGROUND:Many scholars have experimental y confirmed the obvious effect of mesenchymal stem cells to repair radiation injury. OBJECTIVE:To preliminarily investigate the mechanism of human umbilical cord mesenchymal stem cells promoting the healing of combined radiation-wound skin injury and whether they possess tumorigenicity in vitro. METHODS:Fifteen Sprague-Dawley rats were randomly divided into three groups, five rats in each group. The right buttock of rats (2.5 cm×2.0 cm) was irradiated with 40 Gyβ-rays produced by a linear accelerator, in which a round wound with a diameter of 1.5 cm was made. After 12 hours of modeling, human umbilical cord mesenchymal stem cells at three concentrations (5.0×106, 1.0×107 and 2.0×107 ) were injected through tail vein of rats, and luciferin (20 mg/kg) was injected intraperitoneal y. celldistribution in vivo was traced using IVIS in vivo imaging system. The ability of human umbilical cord mesenchymal stem cells to form colonies was observed using the colony formation assay with soft agar. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells injected through tail vein of rats were mostly gathered in the lungs. cells were accumulated in the injured site of rats injected with 2.0×10 7 human umbilical cord mesenchymal stem cells;however, the fluorescence signal was not observed in the injured site of rats injected with 5.0×106 and 1.0×107 human umbilical cord mesenchymal stem cells. The other results indicated human umbilical cord mesenchymal stem cells of low dose, medium dose and high dose had no colony formation on soft agar, but the tumor cells had a great ability to form colony. These findings indicate that human umbilical cord mesenchymal stem cells promote healing combined radiation-wound skin injury by local migration and exhibit no tumorigenicity in vitro in a short period.

BACKGROUND: Recently, a few studies have reported the correlation between transforming growth factor-α (TGF-α) and graft-versus-host disease (GVHD); however, the combination of TGF-α with other cytokines in patients with chronic or acute GVHD requires further study.OBJECTIVE: To analyze the changes of serum tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and transforming growth factor-α (TGF-α) in leukemic patients after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and investigate the effects of these cytokines on different grades of GVHD.DESIGN: Case control study.SETTING: Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA; Department of Nuclear Medicine, the Second Affiliated Hospital, General Hospital of Chinese PLA.PARTICIPANTS: Forty-two leukemic patients (23 males and 19 females, 16-68 years old, mean age of 35 years) who underwent Allo-HSCT for the first time were selected from the Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA and Department of Transplantation, the 307 Hospital of Chinese PLA from June 2005 to June 2007. Twelve patients had acute granulocytic leukemia (AGL), fifteen patients had acute lymphocytic leukemia (ALL), and fifteen patients had chronic granulocytic leukemia (CGL). Among the 42 patients, 37 underwent peripheral blood transplantation and five received bone marrow transplantation. Twenty-one patients had acute GVHD (18 cases in grades Ⅰ-Ⅱ and three cases in grades Ⅲ-Ⅳ) after Allo-HSCT, but the other 21 patients did not. Fourteen patients had chronic GVHD (five cases of limited type and nine cases of extensive type), but the other 28 patients did not. An additional 30 healthy subjects (18 males and 12 females, 20-70 years old, mean age of 44 years) were collected as a normal control group. All patients provided confirmed consent, and the study was approved by the local ethics committee.METHODS: Levels of serum TNF-α, IL-4, and TGF-α in leukemic patients with Allo-HSCT and normal subjects were measured by radio-immuno-assay, the cytokines levels of the patients with/without acute GVHD, of those with/without chronic GVHD and of different grades of GVHD were compared.MAIN OUTCOME MEASURES: Comparisons of serum TNF-α, IL-4, and TGF-α among the groups.RESULTS: All 42 leukemic patients and 30 healthy subjects were included in the final analysis. Levels of TNF-α, IL-4, and TGF-α in patients with acute or chronic GVHD were significantly higher than those in the normal subjects (P<0.05-0.01). Levels of TNF-α and IL-4 in patients without acute GVHD were significantly higher than those in the normal subjects (P<0.01,0.05). Levels of TNF-α, IL-4, and TGF-α in patients with acute GVHD were significantly higher than those in patients without acute GVHD (P<0.05). Levels of TNF-α, IL-4, and TGF-α in patients with chronic GVHD were significantly higher than those in patients without chronic GVHD (P<0.05). Levels of serum TNF-α and TGF-α in patients with acute GVHD of grades Ⅲ-Ⅳ or chronic GVHD of extensive type were significantly higher than those in patients with acute GVHD of grades Ⅰ-Ⅱ or chronic GVHD of limited type (P<0.05-0.01).CONCLUSION: After Allo-HSCT, dynamically monitoring changes of levels of TNF-α, IL-4, and TGF-α may serve as a possible means of predicting the onset of acute or chronic GVHD and may contribute considerably to deciding clinical severity of GVHD.

Objective To investigate molecular epidemiology and antimicrobial susceptibility of Salmonella spp. isolates recovered from the stool samples of children with diarrhea. Methods Seventy-two isolates of Salmonella spp. were collected from children with diarrhea. The serum type of Salmonella spp.was determined by serology agglutinating method. Antimicrobial susceptibility was determined by K-B disk diffusion method and MICs of cefotaxime and ceftazidime were measured by agar dilution method for Salmonella spp. isolates. PCR and DNA sequencing were used for detecting ESBL, ISEcpl and AmpC genes; The transfer of cefotaxime resistance was determined by conjugation experiments. PFGE was performed for determining the homogeneity of the S. typhimurium isolates. Results A total of 72 isolates of Salmonella spp. were collected, among which S. typhimurium accounted for 86 % (62/72) and was the main serum type. S. typhimurium isolates and S. thompson isolates were often resistant to most of clinically used antimicrobial agents. Resistance of S. thompson isolates to ampicillin was the highest (90%, 56/62),followed by tetracycline (81%, 50/62), trimethoprim/sulfamethoxazole (74%, 46/62) and chloramphenicol (66%, 41/62). Seventeen S. typhimurium isolates (27%, 17/62) and two S. thompson isolates were resistant to cefotaxime. Forty-nine S. typhimurium isolates and two S. thompson isolates were positive for blaTEB-1b and resistant to ampicillin. Thirteen ESBL-producing S. typhimurium isolates (21%, 13/62) were positive for blaCTX-M (eight for blaCTX-M-14, three for blaCTX-M-15, one for blaCTX-M-55, one for both blaCTX-M-14 and blaCTX-M-55). All isolates harboring blaCTX-M genes were positive for upstream insert sequence ISEcpl. blaDHA-1was detected in a cefoxitin-resistant S. thompson isolate. Two main clones (PFGE type A and D) accounting for 19% (12/62) and 50% (31/62) respectively were found among 62 S. typhimurium isolates. Seven CTXM-producing isolates belonged to PFGE type D. Conclusions The multi-resistance to antimicrobial agents and high prevalence of blaCTX-M genes are found among S. typhimurium and S. thompson clinical isolates. blaCTX-M-55 is first found in S. typhimurium isolates and blaDHA-1 is found in S. thompson isolates. Clonal spread is responsible for the dissemination of S. typhimurium isolates.

Objective Aimed to clarify the molecular mechanism after subarachnoid hemorrhage （SAH） by investigating the expression of tight junction protein Claudin-5 and ZO-1 and the effects of SP600125 on them. Methods Seventy-five male Sprague Dawley rats （300 to 350 g） were randomly divided into sham,SAH,SAH ＋ DMSO （dimethyl sufoxide） solution,SAH ＋SP600125 （C-Jun N-terminal kinase inhibitor）10 mg/kg,and SAH ＋SP600125 30 mg/kg groups. The standard endovaseular perforation was performed to produce experimental SAH. The JNK inhibitor SP600125 was intraperitoneally administered at 1 hour before and 6 hours after SAH. Results At 24 hours after SAH,signs of microvessels injury were observed in brain cortex. Compared with the sham group,expression of Claudin-5 and ZO-1 was sig- nificantly decreased （P 〈 0.05 ）. JNK inhibitior SP600125 suppressed the decrease of Claudin-5 and ZO-1 expression, attenuated blood-brain barrier disruption in rats after SAH. Conclusions The blood-brain barrier disruption is an important mechanism of early brain injury after SAH. JNK inhibitor SP600125 improves neurological outcomes and provides neuropmtecfion against acute events after SAH such as bloodbrain barrier disruption and cell apoptosis.

MicroRNA has speciifc biological and expression properties in transcription level, which has more potential than genomic DNA in the identification for forensic body fluid, species and degraded crime biological materials. Here we introduce the speciifc research of forensic body lfuid identiifcation, research strategies and applied perspectives with forensic miRNA, expecting to provide the application and study of miRNA analysis for reference.

Objective To evaluate the short-term efficacy of total hip arthroplasty combined with subcutaneous osteotomy in the treatment of CroweⅣ hip dysplasia (DDH).Methods From March 2012 to March 2015,14 patients (16 hips) underwent total hip arthroplasty with femoral distraction osteotomy S-ROM femoral stem prosthesis.And we observed its recent efficacy.Results All patients underwent S-ROM prosthesis.The patients underwent transverse osteotomy of the femoral trochanter.The osteotomy length was 2.0-3.5 cm.The average follow-up time was 19 months.And no complications such as dislocation,vascular nerve injury,deep vein thrombosis and infection were observed during the follow-up.The average Harris scores improved from 42.3 preoperatively to 90.4 postoperatively at 9 months after the operation.The average lengths of preoperative limb shortening and postoperative limb shortening were 6.4 cm and 4.3 cm respectively.The X-ray films showed no dislocation of acetabulum and femoral prosthesis.Bone healing was achieved at 6 months after osteotomy.Conclusion This method could be a good choice for Crowe Ⅳ developmental dysplasia.The short-term efficacy is satisfactory.