Objective To observe the expressions of DNA methyltransferases (DNMTs) 1,3a and 3b in retinoblastoma (RB).Methods Sixty-two RB samples and six normal retinas were studied,including 17 poorly differentiated and 45 well differentiated samples; 16 invasive and 46 non-invasive samples.The expressions of DNMT1,3a,and 3b,and Ki-67 were detected using immunohistochemical analysis.Brown staining of nuclei was considered to represent the positive stain for DNMT1,3a and 3b,and ki-67,blue staining as negative.The level of high expression of nuclear staining was,positive cells in DNMT1≥65 ％,in DNMT3a≥60％ and in DNMT3b≥40％.The correlations of DNMT1,3a and 3b expression in RB samples,and MIB-1 labeling index were analyzed.Results Viewed under the light microscope,negative expressions of DNMT1,3a and 3b were demonstrated in normal retinas,however,positive expression was observed in RB samples,with 100％ in DNMT1,98％ in DNMT3a and 92％ in DNMT3b.Comparing well differentiated RB samples with poorly differentiated samples,significant differences were found in high expression of DNMT1 (x2 =12.57,P＜0.05) and DNMT3a (x2 =10.54,P＜0.05) ; also in the positive cells of DNMT1 (U=179,P＜0.05) and DNMT3a (U=198,P＜0.05).No significant difference was found comparing high expression (x2=1.5,P＞0.05) and positive cells (U=307,P＞0.05) of DNMT3b.When comparing invasive tumor tissues with non-invasive tumors,significant differences were shown between high expression (x2 =4.72,P＜0.05) and positive cells comparing DNMT1 (U=236,P＜0.05).No significant difference was shown in high expression (x2=3.53,0.84; P＞0.05) in DNMT3a and DNMT3b,or in comparison with positive cells (U=338,257; P＞0.05).The expression of DNMTs was positively correlated with the MIB-1 labeling index in RB tissues (R2=0.554,0.376,0.219; P＜0.05).Conclusion There are high expressions of DNMT1,3a,and 3b in RB.

Objective To observe the expression and relationship of high-mobility group A(HMGA) 1,HMGA2,MIB-1 labeling index (LI) and let-7 in retinoblastoma (RB).Methods Forty-four RB samples were studied,including 11 poorly differentiated samples,33 well-differentiated samples; eight invasive and 36 non-invasive samples.The expression of HMGA1,HMGA2 and MIB-1 LI in RB were analyzed by immunohistochemitry.The HMGA1,HMGA2 were scored on a scale of 0 to high expression.0: no expression ; low: 1％ - 10 ％ ; medium: 11 ％ - 50 ％ ; high: ＞50 ％.The MIB LI were scored on a scale of 0to high expression.O: no expression;low: 1％ - 40％;high: ＞ 40％.Semiquantitative reverse transcription-polymerase chain reaction was used to assay the let-7 expression level: ≥ 80％ showed no significantly decreased expression; 60％ - 79％ showed medium decrease in expression; ＜ 60％ highly decreased in expression.Results In 44 RB samples,there were 14 cases with no HMGA1 expression (32％),11 cases with low expression (25％),10 cases with medium expression (23％),and nine cases with high expression (20％).Expression level of HMGA1 was significantly higher in poorly differentiated RB than in well-differentiated RB (x2 =11.3,P＜0.01) ; however,no statistically significant difference was found between invasive tumors and noninvasive tumors (x2 -5.9,P＞0.05).There were 11 cases with no HMGA2 expression (25％),11 cases with low expression-(25％),nine cases with medium expression (20％),and 13 cases with high expression (30％).Expression level of HMGA2 was significantly higher in poorly differentiated and invasive RB than in well differentiated and noninvasive RB respectively (x2=20.9,8.7; P＜0.05).There were 4 cases with no MIB-1 LI expression (9％),18 cases with low expression (41％),and 22 cases with high expression (50％).Expression level of MIB1 LI was significantly higher in poorly differentiated RB than in well-differentiated RB (t=5.2,P＜0.05).Higher expression of MIB-1 LI was found in invasive tumors than in noninvasive tumors,with no significant difference (t=-1.1,P＞0.05).Twenty seven cases had no significantly decreased expression of let-7 (61％).There were eight cases with medium decreased expression (18％) and nine cases with highly decreased expression (21％).Correlation analyses revealed that MIB1 LI expression significantly correlated with HMGA1and HMGA2 proteins (r=0.327,0.602; P＜0.05).A significantly inverse correlation existed between let-7 expression and HMGA1,HMGA2 proteins and MIB-1LI respectively (r=-0.247,-0.310,-0.392; P＜0.05).Conclusions Overexpression of HMGA1,HMGA2 and MIB-1 LI and down regulation of let-7 were demonstrated in RB.Supplying let-7 to RB cells can possibly inhibit HMGA1 and HMGA2 expression.

Adult ; Benzamides ; pharmacology ; Humans ; Hypereosinophilic Syndrome ; drug therapy ; genetics ; Imatinib Mesylate ; Male ; Mutation ; Oncogene Proteins, Fusion ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; mRNA Cleavage and Polyadenylation Factors

Adult ; Anemia, Hemolytic ; chemically induced ; Female ; Humans ; Pancreatitis ; chemically induced ; Pesticides ; poisoning

Objective To detect the clinical features and prognostic factors of nasal cavity malignancy with breast metastasis.Methods 846 Patients with nasal cavity malignancy from January 1999 to December 2011 were enrolled,the clinical and pathological features,clinical diagnostic methods and prognostic factors for breast metastasis patients were analyzed.Results Six female cases (median age 25) were diagnosed with breast metastasis,including 3 rhabdomyosarcoma and 3 olfactory neuroblastoma; consisting 0.7％ of the total 846 cases of primary nasal malignancy group.The metastasis were more likely to be multiple breast lesions with/without metastasis in other site.For the primary tumor,five patients received 66-72 Gy/30-33f of radical radiotherapy,one patient with rest rhabdomyosarcoma received 58 Gy of palliative radiation since breast metastasis was found after 14 Gy of radiation and breast mass resection were performed right after.For the breast metastasis,five of 6 patients received breast surgery,one patients with olfactory neuroblastoma received 6 cycles cyclophosphamide + adriamycin + vincristine chemotherapy.Median survival was 12.7 months.Conclusions For nasal cavity malignancy,breast metastasis more likely occur in younger female patients.Ultrasound may provide useful information in evaluating breast metastasis.Cases combined with metastasis except breast have unfavorable prognosis.

Objective To study the effect of DNA methylation regulation on the toxic effect of paraquat on the sensitized V79 cells t pretreated with 5-aza-2'-deoxycytidine.Methods V79 cells were treated by 5-aza-2'-deoxycytidine (5-Aza-2'-dc) for 12h,which is a DNA methylation inhibitor,and then treated with paraquat for 12h.The morphological changes of V79 cells were observed by microscopy and the cell viability was determined by MTT assay and trypan blue staining method.Results Microscopic examination showed that the combination of 5-Aza-2'-dc and paraquat had stronger effect in inhibiting the growth of V79 cells(the cells became smaller and poorer adhensive ability) than single 5-Aza-2'-dc or paraquat.MTT assay showed that cell viability in the combination group (54.47 ± 3.04) ％ was significantly lower than the 5-Aza-2'-dc group (95.52 ± 0.90) ％ and paraquat group (89.68 ± 4.26) ％ (P＜0.05).Trypan blue staining assay showed that the death rate of ceils in the combination group (53.58 ± 1.57) ％ was significantly higher than the 5-Aza-2'-dc group (7.44 ± 2.31) ％ and paraquat group (12.90 ± 1.21) ％ (P＜0.05) Conclusion 5-Aza-2'-dc promotes V79 cells damage caused by paraquat.

Objective To investigate the immunophenotypic characteristics of acute myeloid leukemia (AML)and its relationship with chemotherapy response.Methods The immunophenotyping was performrd by flow cytometry in 96 newly diagnosed AML patients.Results In 96 patients with AML,the myeloid antigens were mainly expressed including CD13(94/96,97.92％),CD117(86/96,89.58％),CD33(79/96,82.29％)respectively.The positive expression of stem/progenitor cell differentiation antigens were CD38 (92/96,95.83％),HLA-DR(72/96,75.00％)and CD34(60/96,62.50％).While CD34 and HLA-DR were fewly expressed in M3.The CD14 was only expressed in M4 and M5.The lymphocytic antigens had positive expression in the patients with AML,in which CD4 was 37.5％(36/96),CD7 was 22.92％(22/96)and CD2 was 10.42 ％(10/96).In 46 patients for treatment response analysis,total CR rate was 67.39％(31 /46).There was no significant difference of CR rates between the antigens positive group and negative group including CD33,CD117,CD11b,CD14,CD64,CD4,CD34 and HLA-DR.However,the CR rate was significantly lower in CD7 positive cases than CD7 negative ones,with statistically significance difference(P=0.01).Conclusions CD13,CD33and CD117 are the most common antigens in AML.The patients with the positive expression of CD7 have poor response to chemotherapy in AML.

Objective:To Construct 3-D standard external nasal morphological database for nasal prostheses.Methods:12 plaster models of 6 types of external nose were prepared and scanned by a lasser scanner.The data documents were primarily saved as .asc format. The digital noses were partitioned to six areas by the nasal anatomical features: Nasal bridge area,nasal tip and collumella area,left and right dorsal areas,left and right ala nostril areas, the surface model of each area was constructed and connected to a group. With adding the background color, the surface model of external nose was visualized from any observing angles; then the nasal length and width were measured and the original data were replaced by HAN standard data.12 digital nose models with the same size were obtained and the documents were saved by each type with both .asc format and .igs format.Results:Point-cloud data and surface model data of 6 types of standard external digital noses were obtained.Conclusion:The nasal morphological database may be used as the foundation for CAD/CAM technique preparation of nasal prostheses

0.05),20%(P

Objective:To explore the regulatory role of exosomes in calcification of rat vascular smooth muscle cells (VSMC) induced by high phosphorus.Methods:VSMC (A7r5 cells) were cultured in vitro and randomly divided into three groups: normal phosphorus group (0.9 mmol/L), high phosphorus group (2.6 mmol/L) and high phosphorus exosomes induction group (i.e. the exosomes extracted from high phosphorus group were added to VSMC in normal culture). Until the 7th day of culture, the culture medium of normal phosphorus group and high phosphorus group obtained during the change of cell culture medium was collected, and the precipitate was obtained by ultracentrifugation and suspended by phosphate buffered saline. The protein content of the precipitate was determined by BCA protein quantitative method. The precipitates were identified. The structure and size of exosomes were observed by transmission electron microscope. The exosomes marker proteins tumor susceptibility gene 101 protein (TSG101) and CD9 were detected by Western blotting. The miRNA in exosomes was extracted, and the expression of related miRNA (miR-30b, miR-204, miR-211) were observed by real-time quantitative PCR. After 7 days of cultivation, the exosomes uptake process of VSMC in high phosphorus exosomes induction group was observed. The calcium deposition was detected by Alizarin stain, and the calcium content was detected by O-cresol complex copper. The content of alkaline phosphatase was detected by colorimetry. The protein expression of Runx2 was quantified by Western blotting. Results:(1) The precipitate obtained by ultracentrifugation of the cell culture fluid was identified as exosomes by electron microscopy morphology. Western blotting confirmed that the expression of the exosomal marker proteins TSG101 and CD9 were positive. (2) The exosomes were rich in miRNAs. The expression of miR-30b, miR-204, miR-211, which negatively regulated the transcription of Runx2, was significantly down-regulated in the high-phosphorus group compared with the normal group ( P<0.05). (3) After culturing rat VSMC with high phosphorus for 7 days, calcium salt deposition was obvious. Compared with the normal phosphorus group, calcium content and alkaline phosphatase activity were significantly increased (both P<0.05), and Runx2 expression was also significantly increased ( P<0.05). (4) Added the obtained high-phosphorus exosomes to the normal cultured VSMC, the exosomes could be taken up by VSMC and successfully induced VSMC calcification. The levels of cell calcification indicators and Runx2 expression were significantly increased. Conclusions:High phosphorus induces calcification of VSMC and promotes the increase of Runx2 expression. The mechanism may be realized by releasing exosomes from VSMC to transmit cell signals.