Objective To develop a model of be nzene induced aplastic anemia in mice and to observe pathology of aplastic anemi a induced by benzene;Methods we tried to establish a model of benzene induced aplastic anemia in mice,then we studied pathology of aplastic anemia and the pr evention of amifostine on the model.(1) CD1 male mice were injected subcutaneou sly with benzene +corn oil three times a week,and randomly assigned to four gro ups according to the dose of benzene administrated:B1group(0.5ml/kg),B2group( 1.0ml/kg),B3group(1.5ml/kg),B4 group(2.0ml/kg),C group (injected intraperi toneally with 200mg/kg amifostine 30min before benzene administrated on the basi s of the aplastic anemia model as amifostine treated group),while A group treat ed with only corn oil as control,all the groups were complement with corn oil t o 4ml/kg;Before and 2 days after benzene were administrated 10,15,20 and 25 t imes respectively,all the groups mice were killed respectively ,the parameters tested were as following:general condition,peripheral blood smear,histopatho logy of bone marrow 、liver,spleen and PCNA,TUNEL.Results Compared with the control group,some parameters were changed in each model group,but only the pa rameters in B4 group were significantly different from the control group.After 25 times of benzene administration,compared with the control group (A group),B4 group mice showed 18.51% body and 63.86% spleen weight loss (P

Amifostine ; pharmacology ; Anemia, Aplastic ; chemically induced ; pathology ; prevention & control ; Animals ; Benzene ; toxicity ; Dose-Response Relationship, Drug ; Male ; Mice

BackgroundThe study of myopia development is always the hotspot worldwide. Recently,scientist found that some growth factor secreted by retinal nerve epithelium cells and retinal pigment epithelium(RPE)cells are associated with the development of myopia. Whenever, the absorption of RPE cells to different wave-length lights is different. ObjectiveThis study was to investigate the effects of different wave-lengths lights on the proliferation of human RPE cells, and explore the influence of different wave-lengths lights on RPE cells secreting hepatocyte growth factor(HGF) ,basic fibroblast growth factor(bFGF) and transforming growth factor-β(TGF-β).Methods The fourth to fifth passages of human embryonic RPE cells were exposed to blue light( λ =480 nm),red light( λ =775 nm) and white light. The cells of control group were harvested in normal condition. The proliferation and growth of RPE cells were assayed by MTT,and the ultrastructure of cells was examined under the transmission electron microscopy at 48 hours after light exposure of RPE cells. Enzyme linked immunosorbent assay(ELISA) was adopted to determine the concentrations of HGF,bFGF and TGF-β in the culture medium in 12,24,48,72 hours. The expression of HGF mRNA in RPE cells was detected by RT-PCR. This study was approved by Ethic committee of Fudan University. ResultsThe A490 values of the cells exposed to blue light,red light,white light and white light were 0. 0218±0. 0014 ;0. 0353±0. 0025 ;0. 0371 ±0. 0024 and 0. 0445 +0. 0046 respectively with the significant difference among 4 groups ( F =12. 579, P＜0.05 ), and A490 value in blue light group, red light group were significantly lower than that of the control group ( t =2.043 ; t =2.024, P＜0.05 ). ELISA showed that the concentrations of HGF and TGF-β in culture medium were evidently elevated as the prolongation of light exposure in various light exposure groups in 72 hours(HGF) and 48 hours(TGF-β) compared with 12 hours with a predominating rise in the control group. The statistically significant differences were found in the concentrations of HGF and TGF-β between control group and blue light group or red light group in the( all P＜0. 05 ). The bFGF level was decreased with the time increase of various light exposure with the significant differences in 72 hours compared with 12 hours( P＜0.05 ). RT-PCR revealed the considerable difference about expression of HGF mRNA in RPE cells among these four groups( P＜0. 05 ), and the lest expression in HGF mRNA was in the blue light group compared with control group( t =3. 972,P＜0.05 ). Thinning of the chromatin, decreasing of organelle and loss of cellular membrane were seen in the blue light group, but no obvious change of ultrastructure of human embryo RPE cells was found in the ret and white light groups. ConclusionsThe irradiation of different wave-length light can effect the growth and proliferation and secretion of HGF,bFGF and TGFβ in human RPE cells in vitro,implying myopia formation is associated to exposure of different wave-length light.

Objective To investigate the application of real-time three-dimensional transesophageal echocardiography (RT-3D TEE) in quantification of the aortic anuulus diameter in patients with severe aortic stenosis.Methods RT-3D TEE image of aortic root was get perioperatively in 65 patients with sever aortic valve stenosis during the surgery procedure.The aortic annulus was reconstructed in the quantitative analysis software to generate the parameters of 3-dimensional area based aortic anuulus diameter(3D-AAAD) and 3-dimensional circumference based aortic anuulus diameter(3D-C-AAD).The parameters was compared with intraoperative aortic anuulus diameter(IO-AAD),which was get by the standard cylindrical valve sizer.Results The three dimensional form of the aortic annulus was related to the number of the valves.The value of 3D-C AAD and 3D-A-AAD were (22.4 ± 2.1) mm,(23.4 ± 1.9) mm,no significant difference could be detected between them and the IO-AAD (P ＜ 0.05).The 3D-C-AAD had a better agreements with IO-AAD than 3D-A-AAD.Conclusions RT-3D TEE can reconstruct the three dimensional sharp of aortic anuulus,and can be used as quantitative tools to calculate its diameter.

Objective To evaluate the usefulness of real-time three-dimensional transesophageal echocardiography (RT-3D TEE) in the perventricular device occlusion of the ventricular septal defect (VSD).Methods Sixty patients underwent perventricular device occlusion of VSD were divided into two groups.Group A:30 patients,3D images of VSD were obtained intraoperatively by RT-3D TEE.3D images were analyzed and the maximal and minimum area and maximal diameter of VSD were measured in the cardiac cycle.The size of selected VSD occluder was the diameter calculated from the maximal area based on the formula πD2/4 and plus 1～2 mm.Simultaneously,two dimensional transesophageal echocardiography (2D TEE) images of VSD were obtained and measured on standard views.Group B:other 30 patients,intraoperative 2D TEE was performed to measure the diameter of VSD on standard views.The selection VSD occluder was based on the maximal diameter plus 1～2 mm.Results The VSDs shape showed oval,class round and irregular by RT-3D TEE imaging.There was a significant difference between the maximal area and smallest area of VSD appeared in the cardiac cycle obtained by RT-3D TEE (P<0.01).There was a significant difference between the maximal diameter by measurement of RT-3D TEE and 2D TEE (P<0.01).Of all 30 patients with evaluation of VSD size based on the RT-3D TEE,1.1 times attempts of device occlusion were performed on one patient (33/30).Of the other 30 cases with evaluation of VSD size by 2D TEE,2 cases were failed with device occlusion because of instant residual shunt.Of the remaining 28 cases,1.43 attempts of device occlusion were performed on one patient (40/28).Conclusions RT-3D TEE can accurately reveal the size of VSD,and aid in the selection of VSD occluder in the device occluding procedure of VSD.

Objective To study the the feasibility of real-time 3D transesophageal echocardiography (RT-3D TEE) to measure the distance between the coronary ostial and the junction of left ventricle and artery.Methods The RT-3D TEE image of the aortic root was collected intraoperative in 65 cases who were refered to our center for valve replacement.The reconstruction of the aortic annulus was done in the QLAB quantitative analysis software and the height parameter of the annulus could be generated.Meanwhile the coronary artery position was observed and the distance between the coronary ostial and the junction of left ventricle and artery was measured respectively.Results In the 65 cases,49 cases clearly showed bilateral coronary ostial,three cases only showed the left coronary ostial,six cases only showed the right coronary ostial,seven cases of both sides of the coronary ostial were unclear.The shows rate of right coronary ostial was 84.61％ (55/65),with the left coronary ostial 81.54％ (52/65).The measured value of the L-AA in systole and diastole were (15.5 ± 1.9)mm,(12.8 ± 1.5)mm,the R-AA systole and diastole measured values were (13.7 ± 1.8)mm,(12.4 ± 1.7)mm,respectively,there was significant difference between systole and diastole respectively.The aortic annulus height (H) got by 3D reconstruction of the aortic annulus was lower than ventricular-arterial junction to the coronary artery distance in both systole and diastole.Conclusions Not only can RT-3D TEE reconstruct the annulus,but also can show and measure the distance between the coronary ostial and the junction of left ventricle and artery respectively.

Objective To detect the expression of BRAF V600E mutant protein in cutaneous malignant melanoma (CMM),and to evaluate the sensitivity and specificity of immunohistochemistry (IHC) in detecting BRAF V600E mutation.Methods IHC with an anti-BRAF V600E monoclonal antibody was performed to detect the expression of BRAF V600E mutant protein in paraffin-embedded tissue sections from 103 patients with CMM and 40 patients with nevus.Statistical analysis was carried out with SPSS software version 17.0,and the expression rate of BRAF V600E mutant protein was compared by chi-square test.Results The expression rate of BRAF V600E mutant protein in the CMM patients was 20.4％ (21/103),significantly higher than that in the nevus patients (5.0％ (2/40),x2 =5.06,P ＜ 0.05).Significant differences were observed in the expression rate of BRAF V600E mutant protein between CMM patients of different age groups (29.8％ (14/47) in patients aged ＜ 60 years vs.12.5％ (7/56) in those aged ≥ 60 years,P ＜ 0.05) and nationality (30.2％ (13/43) for Uygur nationality vs.13.3％ (8/60) for Han nationality,P ＜ 0.05),as well as among CMM lesions from different anatomical sites (13.6％ (6/42) in acral sites vs.11.8％ (4/29) in mucous membrane vs.45.8％ (11/32) in non-acral sites,P ＜ 0.05) and of different Clark levels (8.6％ (4/42) for grade Ⅰ-Ⅲ vs.12.4％ (17/61) for grade Ⅳ-Ⅴ,P＜ 0.05),but not between male and female CMM patients or between CMM patients with lymph node metastasis and those without (both P ＞ 0.05).IHC with the anti-BRAF V600E antibody showed a sensitivity of 100％ (15/15) and a specificity of 98.5％ (65/66) in detecting BRAF V600E mutation.Conclusions The expression of BRAF V600E mutant protein is up-regulated in CMM lesions,and CMM patients of Uygur nationality seems to have a higher expression rate than those of Han nationality.IHC appears to be an accurate and rapid method to detect V600E BRAF mutation.

Objective To construct and identity pSIREN-HIF-1α/shRNA expression vector in order to make foundation of gene therapy for further exploration of RNA interference to nasopharyngeal darcinoma. Methods According to HIF-1αcDNA gene sequence in the gene bank (NM_001530/NM_181054), a pair of 60 nt oligonucleotides each containing the sites of restriction endonuclease at both ends,were designed and synthesized by Reynolds design principles. Oligonucleotides were annealed and ligated with linedrized RNAi-Ready pSIREN-RetroQ-ZsGreen.Transfected into JM109, the recombinants were finally sequenced and identified by 1%agarose gel electrophoresis. Results The size of the target gene fragment amplified by PCR was 470 bp and in accordance with the expected result.pSIREN-HIF-1α was successfully constructed and identitfied by 1%agarose gel electrophoresis.Sequence analysis of inserted fragment revealed the same sequence as synthesized shRNA Oligonucleotides. Conclusion pSIREN-HIF-1α /shRNA expression vector has been successfully constructed, and can make the foundation of research using liposome packaging transfectiing nasopharyngeal darcinoma cell for the next step .

Amifostine ; pharmacology ; Animals ; Benzene ; toxicity ; Blood ; drug effects ; Blood Cell Count ; Male ; Mice ; Random Allocation

Objective To investigate the effects of different doses of pituitary adenylate cyclase- activating polypeptide(PACAP)on the functional and morphological outcome in a mice model of Parkinson' s disease(PD)rendered by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Methods Male mice were treated with PACAP 0.02, 0.20 or 2.00 ?g by iv bolus for 7 days after MPTP was administered, and were compared with the saline-treated mice.The immunohistochemistry and Western blot were used to detect the alterations of PD biomarker including tyrosine hydroxylase(TH), dopamine transporter(DAT)and vesicular monoamine transporter2(VAMT2).In addition, monoamine neurotransmitters in the striatum of mice were measured by the high performance liquid chromatography (HPLC).Results TH immunohistochemistry indicated that the number of TH-positive neurons in the substantia nigra was increased in all PACAP-treated mice(PACAP(0.02 ?g/d)group was 93.33?4.87, F=85.85,P