1.Passage of superparamagnetic iron oxide labeled rat bone marrow mesenchymal stem cells in vitro: Do iron particles change with passage?
Hua YANG ; Xiaoming ZHANG ; Hong JIANG ; Yang SHAO ; Nanlin ZENG
Chinese Journal of Tissue Engineering Research 2009;13(49):9705-9708
BACKGROUND: As magnetic resonance (MR) contrast, a large number of clinical and experimental researches have been done on superparamagnetic iron oxide (SPIO), while the report on the labeled cell passaged cells is rare.OBJECTIVE: MR imaging was performed to the labeled bone mesenchymal stem cells (BMSCs) and its passaged cells in vitro, in order to establish the base of monitoring magnetic labeled BMSCs with magnetic resonance imaging (MRI) in vivo.DESIGN, TIME AND SETTING: The cytological in vitro experiment was performed at the Imaging Research Institute and Institute of Rheumatology and Immunology of North Sichuan Medical College Hospital from June 2006 to January 2007. MATERIALS: Two clean female albino rats (Animal Center, North Sichuan Medical College), SPIO (Schering AG,Germany) were used in this study.METHODES: Bilateral femur and tibia bone marrow was extracted from rats. BMSCs were harvested and purified using the adherent method, and then labeled with 600 μL ferric oxide-polylysine compound (42 mg/L iron concentration) in vitro. MAIN OUTCOME MEASURES: Cell maker-positive rate and the MR signal intensity were respectively measured to the labeled cells and its passaged cells under the inverted microscope and magnetic resonance imaging. RESULTS: Following Prussian blue staining, labeling rate of SPIO labeled cells at the first passage was 100%. With increased passage, the labeling rate was reduced from the first to fifth passages. Compared with non-labeled PBS control, there was no significant difference in signal intensity in the first and second passages cells, but the signal intensity percentage was gradually decreased with signal intensity of increased cell passage from the third passage. Cell labeling rate was negatively correlated with T2~*WI signal intensity (r=-0.986 6, P <0.005). CONCLUSION: The iron particles in the magnetic labeled cells can be passaged to the offspring cells, and can be monitored in a certain period of time with MRI in vitro. These results firstly introduced that SPIO-labeled cell iron particles can decrease with cell passage.
3.Endoscopic CO2 laser surgery with postcricoid advancement flap for glottic stenosis.
Xiao-Mei SU ; Ping-Jiang GE ; Shao-Hua CHEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2011;46(1):69-70
Adult
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Female
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Glottis
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Humans
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Laryngeal Mucosa
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transplantation
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Laryngoscopy
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Laryngostenosis
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surgery
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Lasers, Gas
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Male
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Middle Aged
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Reconstructive Surgical Procedures
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methods
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Young Adult
4.Superparamagnetic iron oxide labeled mesenchymal stem cells and their magnetic resonance imaging in vitro
Hua YANG ; Xiaoming ZHANG ; Yang SHAO ; Hong JIANG ; Zhaohua ZHAI
Journal of Third Military Medical University 2003;0(17):-
Objective To explore the optimal situation of labeling bone mesenchymal stem cells (BMSCs) with superparamagnetic iron oxides (SPIO) mediated by poly-L-lysine (PLL), and determine the most optimal protocol of magnetic resonance imaging according to the patterns of MR in vitro. Methods BMSCs were isolated from white rat and purified, incubated with SPIO-PLL complexes at the range of concentrations (0, 4.2, 8.4, 21, 42, 84 ?g Fe per ml medium). The labeling ratio and distribution of SPIO particles in BMSCs, and the morphological evidence of abnormal visualization were evaluated by Prussian blue staining, fluorescent microscope and electron microscopy. MTT growth curves and magnetic resonance imagings were obtained at the range of concentrations. Trypan blue exclusion test was performed to elevate the viability of BMSCs labeled with PLL at the range of concentrations (0, 0.05, 0.25, 0.5, 1.0, 5.0 ?g PLL per ml medium). Results The cellular labeling ratio was strongly correlated to the concentrations of SPIO (P
5.The Influence of Thy-1.1 Stem Cell Transplantation on Neointimal Formation of Injured Artery
Shao-Hong DONG ; Hua-Dong LIU ; Xin JIANG ;
Chinese Journal of Hypertension 2006;0(11):-
Objective To study the effect of Thy-1.1 stem cell transplantation on endothelial hyperplasia and restenosis.Methods Thirty 4-6 weeks male SD rats were sacrificed to obtain the Thy-1.1 stem cells.Carotid artery were injured by ballon in sixty female SD rat's were randomized to receive stem cell transplantation(5?10~6 Thy-1.1,n=30)or saline approach(n=30).About 5?10~6 Thy-1.1 stem cells were injected into the injured arter- y after carotid artery injury;while the control rats underwent carotid artery injury and was injected the same amount of saline.The animals were sacrificed,3,7,14,21 and 28 days after balloon denudation.The samples of carotid artery were harvested for pathological examination,RT-PCR and in situ hyhridzation(ISH)were used to detect the transplanted cells in the injured artery.Results The intimal thickness was thinner in stem cell transplantation group(I/M,Stem cell transplantation group:2.06?0.28 vs control group 2.42?0.19,P
6.Role of Matrix Metalloproteinase in Ventilator-Induced Lung Injury Inflammatory Reaction of Newborn Rabbits
shao-dong, HUA ; jiang, DU ; sheng-li, AN ; li-hua, YANG ; zhi-chun, FENG
Journal of Applied Clinical Pediatrics 2006;0(18):-
Objective To investigate the effect of matrix metalloproteinases(MMPs) in ventilator-induced lung injury inflammatory reaction of newborn rabbits.Methods Seventy-five newborn rabbits aged 1-5 days were randomly arranged to 3 groups:control group(n=3),other 72 rabbits designed by 2?2?3 factorial were divided into high concentration oxygen(100%)and low concentration oxygen(45%) groups,and each group was subdivided into 2 groups:High peak inspiratory pressure(PIP)group and low PIP group.All rabbits were done by mechanical ventilation and killed to obtain lung tissue at 1,3,6 hours after trailing respectively.The concentration of MMP-9 and MMP-2 in lung tissue bomogenate were detected by ELISA,at the same time the ratio of wet to dry was detected and pathological section was analyzed.Results 1.The concentration of MMP-2 in high concentration oxygen group,high PIP group and low PIP group increased compared with that in normal group,and there were significant differences.2.The mean concentration of MMP-9 was lower in high concentration oxygen group than that in normal control group,and there was significant difference between 2 groups.The mean concentration of MMP-9 in low concentration oxygen group was higher than that in normal group,and there was no significant difference between 2 groups.There was no significant difference in effect to mean concentration of MMP-9 by different PIP.3.There were positive correlation between MMP-2 and MMP-9,MMP-2 and the ratio of wet to dry in lung.Conclusions Within 6 hours in newborn rabbits by mechanical ventilation,ventilation with high concentration oxygen up-regulate MMP-2,but down-regulated MMP-9.The lungs stretch by mechanical ventilation increase MMP-9,but decrease MMP-2.Mechanical ventilation can affect the synthesis of MMPs,and MMPs plays an important role in ventilator-induced lung injury of newborn rabbits.
7.Analysis of clinical factors related to genotype B and C chronic hepatitis B infection in Zhenjiang area
Chun-Ming LI ; Yu-Hua GONG ; You-Wen TAN ; Jiang-Bo SHAO ; Yuan-Hai ZHANG ;
Chinese Journal of Infectious Diseases 2007;0(11):-
Objective To compare the differences of clinical characteristics between genotype B and C chronic hepatitis B(CHB)patients and to summarize clinical factors related to genotype C hepa- titis B virus(HBV)infection.Methods Seventy eight CHB patients who were diagnosed with genotype B or C infection by liver puncture biopsy and genotyping were enrolled.Their serum HBV DNA levels were detected.Severe hepatitis,liver cirrhosis,hepatocellular carcinoma and HBeAg positive rate were analyzed to determine the pathologic inflammation and fibrosis degree of liver tissue.Chi square test and Logistic multiple regression analysis were employed for the statistical analysis.Results The serum albumin and pre-protein were lower in genotype C CHB patients than that in genotype B.The alanine aminotrans- ferase,total bilirubin and prothrombin time were higher in genotype C CHB patients than that in genotype B.The rates of genotype C patients increased significantly with the grade of liver necroin- flammation progressing from GO to G4(1.8%,11.1%,20.4%,33.3%,33.3%) and the stage of liver fibrosis progressing from SO to S4(5.6%,5.6%,14.8%,33.3%,40.7%),but the rates of genotype B patients did not change significantly with the grade of liver necroinflammation(16.7%, 25.0%,25.0%,20.8%,12.5%)and stage of liver fibrosis progressing(16.7%,29.2%%,20.8%, 16.7%,16.7%).There was statistical significance in grades of liver necroinflammation(X~2= 11.49,P=0.022)and stages of liver fibrosis(X~2=13.56,P=0.006)between genotype B and gen- otype C patients.The rates of genotype C CHB patients were higher than,similar with and lower than the rates of genotype B patients of HBV DNA level above 1.0?10~6 copy/mL,between 5.0?10~2-1.0?10~6 copy/mL and under 5.0?10~2 copy/mL,respectively(51.8% vs 12.5%,35.2% vs 45.8% and 13.0% vs 41.7%).There was statistical significance of HBV loads between genotype B and genotype C patients(X~2=13.25,P=0.001).HBeAg positive rate in genotype C patients was significantly higher than that in genotype B patients(61.1% vs 25.0%,X~2=8.67,P=0.003).The rates of decompensated cirrhosis,compensated cirrhosis and no-cirrhosis in genotype C patients were higher than,similiar with and lower than the rates in genotype B patients,respectively(40.7% vs 4.2%,22.2% vs 20.8% and 37.0% vs 75.0%).There was statistical significance of the rate of cirrhosis between genotype B and genotype C patients (X~2=12.47,P=0.002).Conclusions The degree of liver necroinflammation and fibrosis,the HBeAg positive rate and the incidence of cirrhosis are all related with genotype C HBV infection.
8.Significance of Computed Tomography in Diagnosis of Bronchial Foreign Body in Children
yan, SUN ; shao-hua, WANG ; rong-jun, LIN ; guang-feng, JIANG ; ying, TIAN ; zhi-jun, CHEN
Journal of Applied Clinical Pediatrics 2006;0(16):-
Objective To investigate clinical significance of computed tomography (CT) scan in diagnosis of bronchial foreign body in children.Methods Twenty-one suspected children with bronchial foreign body were studied with spiral CT cross-section scan and coronal reconstruction and diagnosis was confirmed with bronchoscopy.Results The foreign body was displayed in all of 21 cases. CT scan showed foreign body was located in right main bronchial 12 cases, right middle bronchial 1 case, right inferior lobar bronchial 2 cases and left main bronchial 6 cases. Foreign bodies were extracted with bronchoscopy.Conclusion CT scan can display and locate accurately foreign body in bronchial of children,and has very important diagnostic value in patients having atypical histories, clinical and radiological findings.
9.SCREENING AND CHARACTERIZATION OF A PHENOL-DEGRADING BACTERIUM
Li-Hua PAN ; Shao-Tong JIANG ; Peng-Da LIU ; Hui-Xing LI ;
Microbiology 1992;0(05):-
A strain ph 16 , that could effectively degrade phenol,was isolated from sewer sludge of printing and dyeing plant. The preliminary identification sugg ested that the strain belongs to Micrococcus sp. The strain could resist to phenol up to 1.5 g/L. The efficient biodegradation of phenol occurred when th e strain was cultured in the medium (pH 7.0) containing 1.0 g/L phenol under 35℃, wh er e the highest degradation rate reach 99.6% after 36 hours. This strain, when t re ated with some heavy metal ions such as Hg +、Co 2+ and Ag 2+ , showe d the significant inhibition of phenol degradation by 74.2%~100%. The kinetic s of phenol degradation during culture of the strain was also explored.
10.Establishment and optimization of real-time PCR with SYBR Green Ⅰ for quantification of survivin
Li-Hua HU ; Zheng-Jiang CHENG ; Yi-Rong LI ; Shao-Jun HUANG ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.