Objective To investigate the eftect of VEGF gene transtection on endothelial progenitor cell derived from murine bone marrow.Methods Wistar rat's bone marrow was obtained, mononuclear cell isolated,and endothelial progenitor cells(EPS)were cultured in EGM-2MV.EPCs were identified by immunocytochemistry and electron microscope.EPCs were transfected by liposome mediated pcDNA3.0-hVEGF165.VEGF protein level was determined in the cultural medium supernatant after VEGF transfection by ELISA.Cultural medium supernatant was used to co-culture with ECV304,VEGF protein activity was evaluated by MTT.EPCs expression of vWF,VEGF,FLK-1 was detected by immunocytochemistry.Results EPCs were effectively enriched by EGM-2MV,and the EPCs obtained express the typical cell surface markers such as CD34,CD133,FLK-1.The concentration of VEGF protein in supernatant reaches 1280 pg/ml in the 7th day after pcDNA3.0-hVEGF transfection.No influence of EPCs proliferation could be found after transfeetion.The cell surface marker expression of VEGF,FLK-1, vWF became higher with time,and the ratios of positive cell were 88.52%,82.65% and 95.97% respectively.Conclusions pcDNA3.0-hVEGF165 transfeet EPCS mediated by liposome could excrete a high concentration of functional VEGF protein.It is helpful for EPC to maintain the characters of endothelial cell after VEGF gene transfection and differentiate to mature endothelial cell.

Objective To investigate the influence of atorvastatin (Lipitor) on the expression of programmed cell death 4 (PDCD4) in human CD4+T lymphocytes in vitro. Methods Human CD4+T cells obtained from healthy individuals were activated with PHA and treated with atorvastatin. The mRNA and protein expression levels of PDCD4 were detected by real-time PCR and western-blot respectively. Results The stimulation of PHA obviously increased the mRNA and protein expression of PDCD4 and the secretion of those serum cytokines. The expression of PDCD4 and the production of serum TNF-α were significantly decreased, whereas the serum levels of IL-10 were significantly increased after treated by different concentration of atorvastatin. The serum secretion of TNF-α was positive correlation with the expression of PDCD4 through the linear related analysis (r=0.782, P<0.01), and the secretion of IL-10 was negative correlation with the expression of PDCD4 (r=-0.653, P<0.05). Conclusion The anti-inflammatory effects of atorvastatin are mediated by down regulating the expression of PDCD4 in CD4+T cells.

Objective To reconstruct skin defects with local flaps after resection of tumors. Methods From October 2006 to December 2009, medial canthal defects in 15 patients were repaired with local flaps in the hospital. The size of the defects varied from 1 cm × 1 cm to 1.8 cm × 2.0 cm. According to the size and location of the defects, different local flaps such as glabellar flaps ,rotation flaps, advancement flaps, nasolabial flaps and combination of these local flaps were selected and designed to repair the skin defects. Results Of the 15 patients, 12 received the reconstruction of skin defects with single flap, the other 3 with two flaps; totally,18 local flaps were designed and applied. All the flaps survived with primary healing postoperatively. A followup in 11 patients for 6 to 12 months showed that the color and texture of flaps were similar to those of surrounding tissues and incision line scar was inconspicuous. Medial canthal contours were restored without distortion of surrounding structures. Satisfactory function and aesthetic outcomes were achieved. Conclusions In the repair of inner canthus skin defects, to maintain and restore the "cosmetic unit" integrity of inner canthus by using suitable local flaps would lead to a satisfying restoration of function and appearance.

Objective To analyze the clinical features of patients with traumatic brain injury in the Ya' an earthquake and discuss the treatment experiences.Methods Medical records of 69 patients admitted from April 2013 to May 2013 because of traumatic brain injury in the Ya' an earthquake were collected.Retrospective review was performed for age,gender,causes of injury,time from injury to hospitalization,types of injury,associated injury,treatment methods and outcomes.Results There were 47 males and 22 females.Forty-two patients (61％) were injured from falling objects.Fifty-eight patients (84％) were sent to the West China Hospital within 72 hours postinjury.Twenty-two cases (32％) sustained associated injuries.Twenty-nine patients (42％) were critically injured.Twenty-four patients underwent operation at the local hospital and twelve patients had operation at our hospital.Outcome measure using GOS one month after treatment showed 55 favorable recovery,5 moderate disability,4 severe disability,and 5 coma.Conclusions Main cause of injury is hit by falling objects during the Ya' an earthquake.Majority of the patients obtained effective treatment in the time window.GCS in combination with patients' general condition used in casualty triage and critical patients charged by neurosurgeons and treated with the cooperation of multiple disciplinary teams are helpful to successful treatment.

A new thermophilic fungus J18 isolated from the soil samples was identified as Paecilomyces thermophila. This strain produced effectively xylanase utilizing several lignocellulosic materials in the solid-state fermentation (SSF) , and wheat straw was the best carbon source. The results of single-factor-experiment showed that the wheat straw of particle size 0. 3 mm ~ 0.45 mm, initial moisture content of 83% , initial pH of 7. 0 and cultivation temperature of 50℃were the optimal conditions for xylanase production. Under the optimized conditions, it produced 18 580 U/g dry substrate after 8 days of cultivation. Therefore, xylanase production by Paecilomyces thermophila in SSF possess great potential for commercial applications.

Background Researches demonstrated that CpG ODN,a immunostimulatory sequences,has preventing and treating effect on allergic conjunctivitis caused by protein allergen.However,its effect on allergic conjunctivitis caused by fungal allergen is unclear. Objective This study aimed to investigate into whether the Th1-Th2 switching immunostimulatory CpG ODN could reverse the response in the murine allergic conjunctivitis model caused by aspergillus fumigatus. Methods A mixture of spores and hyphae of aspergillus fumigatus strain was used to induce allergic conjunctivitis in male BALB/C mice aged 6-8 weeks.This experiment was designed into preventive or therapeutical treatment program.Under both settings,allergic conjunctivitis of the animals were treated with CpG ODN,nonstimulatory GpC ODN or PBS.After the last challenge with the allergen,the clinical symptoms of the animals were scored based on the criteria of Magone.The animals were sacrificed and the histopathological examination of conjunctiva was performed.Expression of TLR4 mRNA in conjunctiva was analyzed by real-time PCR assay.The responsiveness and populations of lymphocytes in spleen and draining lymph nodes were analyzed by flow cytometry.The use complied with the Standard of Association for Research in Vision and Ophthalmology. Results In the prevention mode.CpG ODN decreased subconjunctival infiltration compared with GpC ODN and PBS groups with the average neutrophil count index(21.25 ±11.59/section,30.75 ±11.44 section and 69.00±9.90/section,respectively).Expression of TLR4 mRNA was up-regulated significantly by CpG ODN.The clinical scores for CpG ODN group were insignificantly lower than those in GpC ODN group and PBS group(P＞0.05).In the therapeutic mode,compared with GpC ODN and PBS groups,the allergic symptom score in CpG ODN group manifested significantly lower(t=4.000.t=2.750,P＜0.01)and showed fewer cellular infiltration(t=4.870,t=3.829,P＜0.01)and higher expression of TLR4 mRNA(P＜0.01).In cultured splenic and draining lymph node cells,increased percentages of CD4+ CD25+ and CD4+ CD25+ CD69+ in CpG ODN group were observed compared with control groups(|P＜0.05). Conclusion CpG ODN can relieve aspergillus fumigatus-induced allergic conjunctivitis via either subconjunctival injection or topical application by upregulating expression of TLR4 and activating Treg lymphocytes.

OBJECTIVETo observe clinical efficacy in treating cutaneous deficiency of traumatic shank amputation stump with full-thickness skin graft combined with vacuum sealing drainage.

METHODSFrom September 2009 to December 2012, 15 patients with cutaneous deficiency of traumatic shank amputation stump were treated with full-thickness skin graft combined with vacuum sealing drainage. Among patients, there were 11 males and 4 females with an average age of 41.5 (ranged from 25 to 62) years old. Ten cases were caused by traffic accident and 5 cases were caused by heavy object, 9 cases on left and 6 cases on right. Six patients with smashed wound were treated with debridement and amputation, combined with vacuum aspiration in-emergency; 9 patients caused by infection and necrosis were treated with debridement and amputation, combined with vacuum aspiration, and full-thickness skin graft were performed at stage II. The skin defect area of residual limbs ranged from 40 cm x 20 cm to 25 cm x 15 cm.

RESULTSAll patients were followed up from 3 months to 1 year. Full-thickness skin graft of residual limbs were survived,and obtained satisfactory walking function with prosthetic. Residual skin increased thicken, wearproof without rupture and pain.

CONCLUSIONFull-thickness skin graft combined with vacuum sealing drainage in treating cutaneous deficiency of traumatic shank amputation stump could reserve the length of residual limbs, increase survival rate of skin graft with less scar of survival skin, get good wearability and it is conducive to prosthetic wear. It is a simple and easy treatment method.

Adult ; Amputation Stumps ; surgery ; Female ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; methods ; Skin ; injuries ; Skin Transplantation

OBJECTIVETo explore the relationship of bone cement distribution and the puncture angle in the treatment of thoracolumbar compression fractures with unilateral percutaneous kyphoplasty (PKP).

METHODSThe clinical data of 37 patients with thoracolumbar osteoporotic compression fractures underwent PKP between January 2013 to March 2014 were retrospectively analyzed, all punctures were performed unilaterally. There were 6 males, aged from 65 to 78 years old with an average of (71.83 ± 6.15) years; and 31 females, aged from 57 to 89 years old with an average of (71.06 ± 7.89) years. Imaging data were analyzed and puncture angle and puncture point were measured before operation. According to the measured data, the puncture were performeds during the operation. Distribution area of bone cement were calculated by X-rays data after operation. The effect of bone cement distribution on suitable puncture angle was analyzed; VAS score was used to evaluate the clinical effects.

RESULTSThe puncture angle of thoracic vertebrae in T8-T12 was from 28° to 33° with an average 30.4°; and the puncture angle of lumbar vertebrae in L1-L5 was from 28° to 35° with an average of 31.3°. Postoperative X-rays showed the area ratios of bilateral bone cement was 0.97 ± 0.15. Bilateral diffuse area were basic equal. Postoperative VAS score decreased significantly (1.89 ± 1.29 vs 7.03 ± 1.42).

CONCLUSIONThrough measure imaging data before operation with PKP,the puncture point and entry point can be confirmed. According the measured data to puncture during operation, unilateral puncture can reach the distribution effect of the bilateral puncture in the treatment of thoracolumbar compression fractures.

Aged ; Aged, 80 and over ; Bone Cements ; Female ; Fractures, Compression ; surgery ; Humans ; Kyphoplasty ; methods ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Spinal Puncture ; methods ; Thoracic Vertebrae ; injuries ; surgery

Objective To investigate the effects of large dose of atorvastatin on the expression of Sprouty-1 in CD4 + T lymphocytes from unstable angina patients during perioperative period of PCI.Methods A total of 52 unstable angina patients enrolled were divided randomly (random number) into large-dose atorvastatin (80 mg/d,n =26) pretreated group and moderate-dose atorvastatin (20 mg/d,n =26) pretreated group.Circulating CD4 + T cells were obtained by magnetic cell sorting system (MACS) before PCI and 18-24h after PCI.For detecting the gene expression,the reverse transcription fluorescent quantitative polymerase chain reaction (RT-PCR) was used to measure the expression of Sprouty-1 mRNA in CD4 + T lymphocyte.The level of Sprouty-1 protein was detected with Western blot analysis and IL-2 was quantified by enzyme-linked immunosorbent assay (ELISA).Results ①Compared with large-dose group before PCI,the expression of Sprouty-1 mRNA and Sprouty-1 protein levels were dramatically increased in large-dose group after PCI (P ＜ 0.05).②Compared with moderate-dose group before PCI,the expression of Sprouty-1 mRNA and protein levels were slightly increased in moderate-dose group after PCI,but there was no statistical significance (P ＞ 0.05).③Compared with large-dose group before PCI,the serum level of IL-2 was decreased in large-dose group after PCI (P ＜ 0.05).Whereas the serum level of IL-2 was slightly increased in moderate-dose group after PCI compared to moderate-dose group before PCI,there was still no statistical significance (P ＞ 0.05).Conclusions Large-dose atorvastatin pretreatment reduced post-PCI myocardial inflammation through up-regulating the expression of Sprouty-1 mRNA and level of Sprouty-1 protein in CD4 + T lymphocytes.

Objective To explore the application of the tracking scan technique in 16-slice spiral CT angiography, in order to improve the quality of CTA. Methods Three hundred patients who were divided into three groups randomly underwent CTA in tracking, testing and estimation scan respectively with GE LightSpeed 16-slice spiral CT. The data of all patients were transmitted to the workstation (AW4.2) and reconstructed. The quality of all images were evaluated by three experienced doctors with double-blind method and divided into four grades (A, B, C and D) from optimal to poor. Results The rate of grade A, B, C and D was 89.00%, 7.00%, 4.00% and 0 respectively for tracking scan, 70.00%, 11.00%, 15.00% and 4.00% respectively for testing scan, while 61.00%, 13.00%, 21.00% and 5.00% respectively for estimation scan. Conclusion Tracking scan technique is superior to testing and estimation scan in image quality of 16-slice spiral CT angiography.