Objective To explore the relation ship between expression of survivin gene in leukemia cells and its clinical effects, and to study the mechanism of survivin resist-apoptosis function in leukemia.Methods Survivin expression was detected by Western blots analysis and expressions of Fas and Caspase were examined by immunohistochemistry in 18 leukemia patients.Results Thirteen cases in peripheral blood mononuclear cell survivin positive expression was detected in 18 leukemia patients(72.2%), but no survivin expression in 10 normal persons. There were significant difference of survivin expression in ALL/ANLL patients groups and different WBC groups(P

Objective To investigate the effects of recombinate mouse granulocyte-macrophage-colony-stimulating factor(GM-CSF) alone with mesenchymal stem cells(MSCs) on injured lung of rots after exposure to hyperoxia. Method Mouse MSCs were separated, cultured, amplificated, identified and labeled with 4', 6-diamidino-2-phenylindole(DAPI). Thirty-two 3-day-old Sprague Dawley(SD) rats from four litters were randomly divided(random number) into four groups, namely hypemxia exposured + GM-CSF + MSCs group(group A), hyperoxia exposured + GM-CSF group( group B), hyperoxia exposured + MSCs group( group C) and hypemxia exposured group(group D). All rats were placed in a closed Plexiglas chamber with a minimal flow in and out, providing six to seven exchanges of the chamber volume per hour and maintaining O_2 levels above 95%. Seven days lair,all of them were taken out of the chamber. Rats in group A were treated with 5 x 10~4 MSCs intraperitoneally alone with 9 μg/kg GM-CSF subcutaneously, rats in group B received 9 μg/kg GM-CSF subcutaneously, rats of group C were treated with 5 x 10~4 MSCs intraperitonealiy and rats of group D were administrated with phosphatebuffered solution(PBS). Three days latter, all animals were sacrificed by an injection of 120 mg/kg sodium pentobarbital, perfused with cold 0.9% NaCl, and the left lungs were removed. The upper lobe of them were grinded to make tissue homogenates used for ELASA, and the lower lobes of them were made into frozen sections for fluorescence microscope. The right lungs were fixed in situ for 2 hours by intratracheal instillation of 10% neutral formalin and then were still fixed for additional 24 hours. Sagittal sections of paraffin-embedded middle lobe and upper lobe of left hmg tissue were used for Immunohistochemistry and histological study respectively, Immunohistochemistry was used to detect the expression of telomerase reverse transcrip tase(TERT) grade. Results Among 4groups, there were significantly differences in radical alveolar counts(RAC), TNF-α and IL-β1 in tissue homogenates( P < 0. 01 ). Compared with group D, increase in RAC and the decrease in both TNF-α and IL-β1 were found in other groups, and furthermore there were obvious differences in those changes between group A and group B as well as between group A and group C. There were significant differences in TERT(P<0.01) among four groups. The TERT grade in group A and group B were increased more markedly. DAPI-positive cells were found in group A and group C with significantly differences(6.23 ± 1.88, 5.10 ± 0.91, t = 1.53, P<0.05).Conclusions The protective effects of GM-CSF/MSCs on injuried lungs of new born rats after exposure to hyperoxia may be associated with the increase in the proliferation of stem cells improving the local micro-environment of lung tissue. This protective effects against lung injury induced by hyperoxia exposure may be attributed to the synergism between GM-CSF and MSCs.

Objective To explore the relationship between bronchial asthma severity and the levels of nerve growth factor(NGF) in induced sputum.Methods One hundred and seventy children with asthma were divided into two groups, including 87 cases of chronic duration group and 83 cases of acute phase group.Before treatment lung function test was conducted, induced sputum was taken immediaitely, the levels of NGF were measured by enzyme-linked immunosorbent assay and Eosinophils (EOS)in induced sputum were count.Comparative analysis was conducted.Results The levels of NGF,FEV1 ,FVC,PEF and EOS in duration group were significantly elevated compared with acute phase group (all P ＜ 0.05).Significant differences in NGF, FEV1, FVC, PEF and Eos were found by comparison of different extent in chronic duration group and acute phase group (all P ＜ 0.05), the levels of NGF were positive correlated with illness severity (all P ＜ 0.05) in children with afsthma in chronic duration and acute phase, the levels of NGF were positive correlation with FEV1, FVC, PE-Fand EOS (all P ＜ 0.05) in children with asthma in chronic duration, the levels of NGF were positive correlation with FEV1 and PEF in children with Asthma in acute phase,but the levels of NGF has no correlation with FVC and EOS (all P ＞ 0.05) in children with Asthma in acute phase.Conclusion The levels of NGF in induced sputum of children with asthma in acute phase are markedly elevated compared with children with asthma in chronic duration, the levels of NGF are positive correlated with illness severity in children with asthma in chronic duration and acute phase.

Objective In clinical practice,ultrasound-guided fine-needle aspiration biopsy (US-G FNAB) is the gold standard in diagnosing the pathological nature of undetermined thyroid nodules before operation.Re---sults of FNAB may vary substantially among operators with different skills.The aim of this study is to evaluate whether operator experience in US-G FNAB influences the rate of diagnostic procedures.Methods A total of 600 consecutive US-guided FNAB done by a single radiologist with 22-or 27-gauge needles from Feb.2008 to Apr.2012 were retrospectively analyzed.All specimens were prepared and fixed without the cytologist on site and were subsequently analyzed by 2 expert cytologists.The procedures were chronologically divided into 6 groups and further stratified according to the overall (A),solid nodules (B),mixed nodules (C),pure cystic nodules (D),acellular sample(E),heavily blood-stained sample(F),exclusively colloid material(G),incorrect slide fixation(H) and they were classified as diagnostic or no-diagnostic.Results The rate of no-diagnostic procedures for each group in learning curves were reported as:① Among A,34％ in group 1,15％ in group 2,18％in group 3,10％ in group 4,7％ in group 5,and 8％ in group 6.② Among B,4％ in group 1,1％ in group 2,0％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.② Among C,10％ in group 1,1％ in group 2,4％ in group 3,0％ in group 4,1％ in group 5,and 0％ in group 6.④ Among D,20％ in group 1,13％ in group 2,14％ in group 3,10％ in group 4,6％ in group 5,and 8％ in group 6.⑤ Among E,25％ in group 1,15％ in group 2,15％ in group 3,10％ in group 4,6％ in group 5,and 8％ in group 6.⑥ Among F,4％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.⑦ Among G,1％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,1％ in group 5,and 0％ in group 6.⑧ Among H,4％ in group 1,0％ in group 2,1％ in group 3,0％ in group 4,0％ in group 5,and 0％ in group 6.Among the 600 procedures,there were 2 cases of vertigo and 4 cases of moderate bleeding at the biopsy site,all of which resolved spontaneously.No major complications were recorded.Conclusions The rate of no-diagnostic US-guided FNAB is heavily dependent on the operator's experience.We estimate that at least 200 procedures are needed in order to achieve the levels of diagnostic accuracy reported in the literature.We therefore suggest operators to get specific training before routinely perform this procedure in clinical practice.

Objective To evaluate the clinical application of sweep pattern visual evoked potential in assessing visual acuity (SPVEP-A)of amblyopic children.Design Prospective clinical study.Participants 80 patients(148 case)of amblyopic children and 26 children(52 case)with normal vision.Methods The correlation were analysed of SPVEP-A and E acuity in 80 amblyopic children and of SPVEP-A and PVEP-A in 26 children of normal visual.SPVEP-A was recorded with UTAS-E 3000(LKC)using gratings of 25 differ- ent spatial frequency from 0.2 to 18.8cpd as stimulus.The response were averaged and DFT on the monitor display.SPVEP-A was de- termined by extrapolate to 0 response amplitude.Both transient PVEP-A and SPVEP-A were examined in 26 normal children.The ob- jective visual acuity in PVEP were determined with the highest spatial frequency which evoked a recognizable response.Main Out- come Measures SPVEP-A,PVEP-A and E acuity.Results SPVEP-A were from 0.35 to 0.9 among 80 patients with corrected E acu- ity from 0.1 to 0.8.The correlation coefficient of SVEP-A-I with E acuity was 0.602 .The correlation coefficient of SPVEP-A.between reproducibility detection was 0.448.The correlation between SPVEP-A and E acuity in amblyopic slightly was higher than middle and severe amblyopia(r=0.773 vs r=0.590).In 26 normal children a correlation coefficient of 0.679 was obtained between E acuity anti PVEP-A but 0.424 between E acuity and SPVEP-A.Higher evaluation or lower evaluation of E acuity was found in SPVEP-A in this study.Acuity was upostimated in lower scope of E acuity and underestimated in higher scope of E acuity.The reproducibility and vari- ation were not satisified.Conclusion The sweep VEP is a objective method of amblyopic screening.There is significant positive corre- lation between SPVEP-A and E,but its variability is more great,so we should improve its stability and positive rate uheriorly.

In order to observe the replication and expression of HGV RNA genome in HepG2 cells and establish a cell model of HGV infection, HGV RNA genome was prepared in vitro and transfected HepG2 cells with lipofec-tamin. HGV RNA-positive supernatants were used to infect fresh HepG2 cells. RT-PCR, immunohistochemistry and Western blot assays were carried out to detect the replication and expression of HGV in HepG2 cells. Both positive and negative strands of HGV RNA could be detectable in cell culture supernatants and cells at 24h post-transfection. During the culture periods of 90 days, the cells were maintained by changing the medium every 3 or 5 days, and cultured for more than 20 passages. Both strands of HGV could be detectable in culture supernatants and cells. Immunohistochemistry and Western blot results also confirmed that HGV E2 protein could be expressed in the infected HepG2 cells. HGV RNA could also be detectable in the frozen-thawed HepG2 cells infected with HGV RNA genome. Therefore, HGV RNA genome can replicate and express in HepG2 cells, this HGV RNA genome transfected cells model could be used as a cell model in the studies of replication and infection of HGV.

Objective To compare the anti-inflammatory effects between methotrexate(MTX) and Yangqixue Qufengshi Recipe(YQXQFSR) on mice arthritis induced by collagen II.Methods The model of rheumatoid arthritis was induced by using collagen II in C_(57)BL/6?DBA/1 mice.The rats with arthritis were treated with MTX or YQXQFSR respectively.The indexes such as the onset day of collagen induced arthritis(CIA),the level of CII-reactive antibodies and the pathological scores of CIA were assessed.Results The onset day of CIA and the level of CII-reactive antibodies were not influenced in the treatment of mice arthritis with MTX and YQXQFSR respectively.But MTX could alleviate the damage of cartilage [(1.60?1.51) vs(3.56?1.33) scores,P

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2％ trypsin and 0.133％ collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1％ fibronectin and cultured in endothelial cell-specialized medium supplemented with 10％ fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.