Adult ; Humans ; Male ; Neck Injuries ; complications ; Pharynx ; injuries ; Trachea ; injuries

AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.

Objective To establish a new method for rapid detection of electrolytes based on micro spectrometer and dry reagent.Methods The liquid electrolyte reagent was lyophilized for the preparation of reagentin powder form,which was then sealed into a detector cup of micro spectrometer for later experiment.During determination,the detector cup,in which the specimens and diluents were added to the dry reagent,was put into the detector slot of micro spectrometer,the contents in the detector cup were then well mixed by the magnetic stirring system of micro spectrometer and incubated for 6min with the heating system.Afterwards,the A values of potassium ion,sodium ion and chloride ion were detected respectively at 620nm,405nm and 456nm following the reaction principles of turbidimetry,enzymatic method and chemical method.Based on the findings of those detections mentioned above,the performance of the electrolytic dry reagent was evaluated,and the results were then statistically analyzed.Results The linear range of each parameter could meet the demand for clinical analysis,and the dry reagents had good reaction stability for 90 days after being lyophilized,with the intra-assay coefficient variation(CV) less than 4%,inter-assay CV less than 5%,and the recovery rate from 95% to 105%.No obvious interference was observed in the determination results under the circumstance that the serum TB was less than 290.40?mol/L and the TG was less than 11.20mmol/L.The detection results by this method were well correlated with that of dry chemical analysator VITRO S-250(R≥0.98).Conclusion The method is accurate and reliable in determining the serum electrolytes,and its process is simple process and convenient to carry out.Therefore,it can satisfy the condition for field aid as well as primary care.

Objective To evaluate the effects of percutaneous embolectomy for treament of acute superior mesenteric artery embolism.Methods 34 cases(atrial fibrillation 14 cases;left atrium myxoma 2 cases;SMA thrombosis 15 cases and chronic mesenteric ischemia 3 cases)of acute mesenteric artery embolism were treated with percutaneous embolectomy using long sheath aspiration method and/or thrombolysis via catheterization of the SMA.Results Successful recanalizations were achieved in all of the 34 superior mesenteric arterial embolism including recovery in 31 cases,laparotomy for 2 cases,and 1 died within 24 hours.Conclusions Percutaneous embolectomy using long sheath aspiration is a simple and effective method for treatment of acute superior mesenteric artery embolism.A correct pathogenetie diagnosis is the key to improve the curative effect and avoid the severe complication.(J Intervent Radiol,2005,15:206-208)

Objective To evaluate the clinical effect of interventional therapy in treating intestinal ischemia of mesenteric venous thrombosis.Methods Twelve cases(male 7 cases,female 5 cases;ranging from 33 to 86 years of age)of mesenteric venous thrombosis(MVT)were treated with percutaneous transhepatic mesenteric venous thrombectomy and thrombolysis associated with papaverin perfusion via superior mesenteric artery.Results Seven of the 12 cases recovered;3 cases were undertaken laparotomy;2 died within 30 days respectively.No severe complications occurred in all of the 12 cases.Conclusions Interventional therapy of MVT is a safe and effective method with reduction of the mortality.(J Intervent Radiol,2006,15:202-205)

Background Experimental study showed that heparanase (HPA)is overexpressed in choroidal neovascularization,suggesting that it may play a role in the pathogenesis of angiogenesis.To certify HPA inhibitor suppress the formation and development of new blood vessel has an important significance for the treatment of choroidal neovascularization.Objective The aim of this study was to investigate the effect of HPA inhibitor on the proliferation of human umbilical vein vascular endothelial cell (UVEC) and the expression of HPA.Methods Hunan UVEC was primarily cultured and passaged and the third generation cells were used in the experiment.Phosphomannopentaose sulfate (PI-88) solution,a HPA inhibitor,was prepared with endothelial cell medium and the end concentrations were 400.00,200.00,100.00,50.00,25.00,12.50,6.25 mg/L respectively.The cells were treated with PI-88 solutions for 24,48 and 72 hours.The growth and proliferation of human UVEC were analyzed using MTT colorimetric assay at absorbance 570 nm.The expression of HPA in the cells was detected by immunochemistry in 48 hours after addition of PI-88.Results Cultured human UVEC showed the fusiform and polygon in shape.The A570 values of human CVEC were significantly different among various concentrations of PI-88 groups (F=2.721,P=0.053 ) and different action time (F=9.656,P =0.002).When PI-88 was administered for 24 hours,the A570 values of human UVEC were insignificantly altered in comparison with the one without PI-88 culture group (P＞0.05 ).However,in 72 hours after experiment,the A570 values were significantly declined as the PI-88 concentration was ＞50.00 mg/L ( P＜0.05 ).When PI-88 was administered for 48 and 72 hours,the A570 values of human UVEC were significantly higher than those of 24 hours in ＜50.00 mg/L groups (P＜0.01 ),but no statistical differences were seen in ＞100.00 mg/L groups among various time points (P＞0.05 ).HPA was intently expressed in the cytoplasm of human UVEC.However,at 48 hours after addition of ＞25.00 mg/L PI-88,the HPA expression was obviously weaker.Conclusions PI-88 can suppress the growth and proliferation of human UVEC at the dose-and time-dependent manner by downregulating the expression of HPA in the cells.

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.

Animals ; Female ; Male ; Rabbits ; Tumor Necrosis Factor-alpha ; analysis ; Vibration ; adverse effects

Objective To investigate the expression of T cell immunoglobulin domain and mucin domain (TIM)-3 and its ligand Galectin-9 in the peripheral blood of initial systemic lupus erythematosus (SLE)patients,and explore their effects on SLE.Methods The percentages of CD4+TIM-3+,CD8+TIM-3+cells from 33 SLE patients and 26 normal controls were detected by flow cytometry,and the Galectin-9 gene expression of PBMCs was determined by real-time PCR The SLE Disease Activity Index(SLEDAI),C3 level and lymphocyte count were evaluated.Mann-Whitney U test was used for independent samples analysis and Spearmen's test was used for correlation analysis.Results The percentages of CD4+TIM-3+ and CD8+TIM-3+ cells were markedly increased in SLE group than those of the control group(P<0.01).In particular,the CD4+TIM-3+,CD8+TIM-3+ level Was positively correlated with SLEDAI (r=0.517,P＜0.01;r=0.400,P＜0.05);but negatively correlated with C3(r=0.487,P＜0.05;r=0.395,P＜0.05).The Galectin-9 mRNA in SLE PBMCs was higher than that of the controls(P<0.05).Conclusion TIM-3-Galectin-9 pathway may be involved in T cell immune regulation of SLE,and is related to disease activity.

Humans ; Medicine, Chinese Traditional ; methods ; Tongue ; metabolism ; pathology