r diagnosing the disease to combine pathology, immunohistochemistry and SYT-SSX gene detection.

Objective To analyze the prognostic factors in patients with cervical squamous cell carcinoma of stage Ⅰb and Ⅱa treated by surgery,and to investigate their guid roles in available post-operation adjuvant therapy. Methods The clinicopathologic records of 306 patients with cervical squamous cell carcinoma of stage Ⅰb and Ⅱa who underwent radical hysterectomy and pelvic lymphadenectomy were retrospectively analyzed, and the prognostic factors were explored by univariate and multivariate methods. Independent prognostic factors were identified by COX proportional hazards regression model. Results The overall 5-year survival rate of these 306 patients was 78.1%. In univariate survival analysis, the poor prognostic factors included poor differentiation, positive pelvic lymph nodes, deep stromal invasion, parametrial extension, tumor size≥4 cm, and lymph vascular space involvement (P

Objective To evaluate the relevance of MSCT pulmonary function parameters and pulmonary function test (PFT) parameters, and to define the reference value of MSCT pulmonary function parameters. Methods Thirty male volunteers received clinical PFT and MSCT scan. MSCT scan was perfomed at the end of the maximum inspiratory and maximum expiratory. All data were analyzed with the lung analysis software of computer-aided inspection system correlatedly with pulmonary function parameters. Results The lung volume at full inspiratory volume (Vin) and full expiratory volume (Vex) in MSCT scan had good correlation with total lung capacity (TLC) and residual volume (RV) (r=0.90, P<0.01; r=0.74, P<0.01). Vex/Vin was correlated with RV/TLC (r=0.74, P<0.01), and Vin-Vex was correlated with MVC (r=0.85, P<0.01). In inspiration, the average lung density was (-879.51±32.82) HU, the density per unit volume was (0.12±0.03) g/cm3, while in expiratory they were (-688.14±62.38) HU and (0.31±0.06) g/cm3. Conclusion MSCT pulmonary function tests with the analysis software of computer-aided inspection system have good correlation with PFT.

ObjectiveTo evaluate the clinical feasibility of low dose MSCT for quantitative assessment of pulmonary function in smokers.MethodsOne hundred and forty-six patients with chronic objective pulmonary disease ( COPD ) including 109 smokers ( 74.6％ ) and 37 non-smokers ( 25.3％ )underwent pulmonary function test and low-dose MSCT scan.All data were analyzed using computer-aided lung anlysis software.Pulmonary function parameters from low-dose MSCT were compared between smokers and non-smokers and also compared with pulmonary function test in non-smokers ( Pearson test).Results In smokers,the average volume at full inspiratory phase (Vin) was (5125 ± 862 ) ml,mean lung attenuation was ( - 902 ± 26 ) HU,mean lung density was (0.0984 ± 0.0260 ) g/cm3,emphysema volume was (2890 ±1370) ml.The average volume at full expiratory phase (Vex) was (2756 ±1027) ml,mean lung attenuation was ( -811 ±62) HU,mean lung density was (0.1878 ±0.0631 ) g/cm3,emphysema volume was (685 ±104) ml.In non-smokers,the average Vin was (3734 ± 759) ml,mean lung attenuation was ( -876 ±40) HU,mean lung density was (0.1244 ±0.0401 )g/cm3,emphysema volume was ( 1503 ± 1217) ml.The average Vex was ( 1770 ± 679 ) ml,mean lung attenuation was ( - 765 ± 56 ) HU,mean lung density was (0.2360 ± 0.0563 ) g/cm3,emphysema volume was ( 156 ± 45 ) ml.There were significant differences between smokers and non-smokers (P ＜0.01 ).The Vex/Vin was correlated with residual volume/total lung capacity ( RV/TLC,r =0.60,P＜0.01 ),and Vin was correlated with TLC ( r =0.58,P ＜ 0.01 ),Vex with RV ( r =0.59,P＜0.01 ).Pixel index ( PI ) -950in was correlated with FEV 1％ pre and FEV1/FVC％ ( r =- 0.53,- 0.62,respective,P ＜ 0.01 ),Pl-950ex was correlated with FEV1 ％ pre and FEV1/FVC％ ( r =-0.71,-0.77,respective,P＜0.0l).ConclusionLow-dose MSCT can be a potential imaging tool for quantitative pulmonary function assessment in smokes.

OBJECTIVETo establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis.

METHODSRun immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized.

RESULTSThe 2-DE patterns of uterine leiomyoma and myometrium with good quality were obtained.

CONCLUSIONWith optimal condition the two-dimensional electrophoresis of uterine leiomyoma can be obtained.

Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leiomyoma ; chemistry ; Myometrium ; chemistry ; Neoplasm Proteins ; analysis ; Proteome ; analysis ; Uterine Neoplasms ; chemistry

OBJECTIVETo establish a methotrexate (MTX)-resistant choriocarcinoma cell line and to determine its biologic characteristics.

METHODSMTX-resistant cell line (JAR/MTX) was derived from human choriocarcinoma cell line JAR by exposed to intermittently and progressively increasing concentration of MTX. Drug sensitivity was detected by MTT; P-gp GST-Pi and PCNA expressions were detected by immunohistochemistry. Cell apoptosis was detected by flow cytometry (FCM) with PI/Annexin V stain. Growth rates and human chorionic gonadotropin (HCG) production were also measured.

RESULTSJAR/MTX cell line was established with stable MTX-resistance (resistance index to MTX was 7.3) and cross-resistant to TAX and VCR. Growthrate of JAR/MTX was lower than that of parent cell line JAR. Expression level of PCNA in JAR/MTX was lower than that in JAR (3.09+/-0.42 compared with 3.72+/-0.35, P<0.05), while GST-pi expression was higher. No statistical difference of P-gp expression existed between two cell lines. JAR/MTX secreted more HCG than JAR every 10(5) cells secreted (95.7+/-5.4 compared with 41.3+/-2.8)mIU after 48 h(P<0.01). The flow cytometry showed that the spontaneous and MTX induced apoptosis in JAR/MTX was significantly lower than that in JAR P<0.05.

CONCLUSIONJAR/MTX cell line presented stable resistant to MTX and cross-resistant to TAX and VCR, which might sever as a model in study of drug resistance in choriocarcinoma.

Annexin A5 ; analysis ; Apoptosis ; Cell Adhesion ; Cell Division ; Cell Line, Tumor ; Choriocarcinoma ; drug therapy ; genetics ; pathology ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology

OBJECTIVETo study the expressions of VEGF/VEGFRs and activation of STATs in ovarian epithelial carcinoma, and to elucidate direct effect of VEGF on ovarian carcinoma cells.

METHODSTissue samples from 42 women with primary ovarian epithelial carcinoma (OVCA), 29 with begnin ovarian tumor (OVBT) and 11 with normal ovarian tissue (NOV) were collected. LSAB immunohistochemical staining was used to determine the expression of VEGF, VEGFR1, VEGFR2 and activated STATS (P-STAT1, P-STAT3, P-STAT5, P-STAT6) proteins.

RESULTS(1) Semi-quantitative scoring showed that VEGF expression in OVCA was significantly higher than that in OVBT and NOV (P < 0.01). Expressions of VEGFR1 and VEGFR2 were significantly elevated in OVCA, including tumor cells and stromal vascular endothelial cells (P < 0.01, compared with OVBT and NOV). There was no difference in VEGFRs expressions between OVBT and NOV. (2) In OVCA, tumor cells and endothelial cells expressed P-STAT3 and P-STAT5 at significantly higher levels than those in OVBT and NOV (P = 0.000). The staining of P-STAT1 and P-STAT6 was weak with no significant differences among OVCA, OVBT and NOV. (3) Expressions of VEGFR1 and VEGFR2 in endothelial cells were significantly correlated with P-STAT5 and P-STAT3, respectively (P = 0.006 and 0.001). In cancer cells, VEGF, VEGFR1 and VEGFR2 were all significantly correlated with P-STAT3 and P-STAT5 (P = 0.000), but not with P-STAT1 or P-STAT6.

CONCLUSIONVEGF affects ovarian carcinoma cells via VEGFRs, and STATs probably participate in intracellular signaling of VEGF.

Adult ; Aged ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Cystadenoma, Mucinous ; metabolism ; pathology ; Cystadenoma, Serous ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Endothelial Cells ; metabolism ; Female ; Humans ; Middle Aged ; Milk Proteins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; Ovary ; metabolism ; STAT3 Transcription Factor ; STAT5 Transcription Factor ; Signal Transduction ; Trans-Activators ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo investigate the expression of OPCML gene in ovarian epithelial carcinoma and determine the relationship between mRNA expression and methylation of their promoters.

METHODTwenty normal ovarian tissues and 89 ovarian epithelial tumor specimens (72 malignant, 17 benign), as well as 3 ovarian carcinoma cell lines (SKOV-3, CAOV3, and 3AO), were collected for detection of OPCML gene expression by reverse transcription-polymerase chain reaction and for detection of promoter methylation by restriction enzyme cut analysis from 7. 1999 to 7. 2003.

RESULTSAmong ovarian epithelial carcinoma 19.4% expressed OPCML mRNA, while 85% of normal ovarian tissue and 76.5% of benign ovarian tumor. The ratio of expression of OPCML mRNA in ovarian epithelial carcinoma was significantly lower than those of normal (chi2 = 30.108, P = 0.0000) and benign tumors (chi2 = 21.162, P = 0.000). No OPCML mRNA expression was found in SKOV-3 and CAOV3, but was found in 3AO. Methylations were detected in 44.4% of cancer cells promoter, while 0% in normal ovarian tissue and benign ovarian tumors. The ratio of methylation of ovarian epithelial carcinoma was significantly higher than those of normal (chi2 = 13.630, P = 0.0000) and benign tumors (chi2 = 11.797, P = 0.000). Methylation was found in SKOV-3 and CAOV3, but not in 3AO. The relationship between gene expression and promoter methylation was correlated (r = 11.589, P = 0.002), especially at Hap I1 site (r = 11.640, P = 0.004). Methylation was also found in SKOV-3 and CAOV3 cell lines, but not in 3AO cell line.

CONCLUSIONDeletion of OPCML gene exists in ovarian epithelial carcinoma cell. The gene promoter methylations, especially Hap II motif, may be one of pathways that contribute the inhibition of OPCML expression.

Adult ; Aged ; Cell Adhesion Molecules ; genetics ; Cell Line, Tumor ; CpG Islands ; genetics ; DNA Methylation ; Female ; GPI-Linked Proteins ; Gene Deletion ; Humans ; Middle Aged ; Ovarian Neoplasms ; genetics ; pathology ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the activation pattern of signal transducers and activators of transcription (STAT) induced by vascular endothelial growth factor (VEGF) in CD34+ hematopoietic progenitor cells, and gain an insight into the molecular mechanism and signal transduction pathway of VEGF that has an effect on CD34+ hematopoietic progenitor cells.

METHODSAfter isolated from umbilical cord blood by using a high-gradient magnetically activated cell sorting system (MACS), CD34+ cells were stimulated by VEGF (50 ng/ml) for different time (0, 15, 30, 45, 60, 90 min) to detect the tyrosine phosphorylation and nuclear translocation of STAT-3 and STAT-5 with Western blot and immunocytochemistry methods. The expression of VEGF receptor-2 (VEGFR2) on the membrane of CD34+ progenitor cells was examined by immunocytochemistry. ATWLPPR, an effective peptide screened from phage epitope library by affinity for membrane-expressed VEGFR2 and blocking the binding of VEGF to VEGFR2, was used to determine whether the activation of STAT pathway induced by VEGF was blocked.

RESULTSTyrosine phosphorylation of STAT-3 and STAT-5 was undetectable in unstimulated CD34+ cells, but was evident at 15 min in response to VEGF stimulation. VEGF resulted in a rapid and transient tyrosine phosphorylation of STAT-3 and STAT-5. The maximal tyrosine phosphorylation was catched at 30 and 45 min, respectively (P = 0.0001), and returned to basal levels at 90 min. Immunocytochemistry confirmed that increased phosphorylated STAT-3 was translocated into the nuclei at 30 min (P = 0.0001), and mainly in cytoplasms again at 90 min after stimulation with VEGF. However, compared with unstimulated CD34+ cells, there was only increased phosphorylation of STAT-5 appeared mainly in cytoplasms, but no significant nuclear translocation was found after stimulation with VEGF (P > 0.05). The presence of VEGFR2 was confirmed using anti-VEGFR2 antibody staining by immunocytochemistry, moreover, the phosphorylation of STAT-3 and STAT-5 failed to be activated by the co-culture with ATWLPPR and VEGF, suggesting that activation of the STAT pathway be specifically mediated by VEGFR2 in CD34+ progenitor cells.

CONCLUSIONSSTAT signaling pathway participates in the signal transduction of VEGF via VEGFR2 in CD34+ hemopoietic progenitor cells.

Adult ; Antigens, CD34 ; metabolism ; DNA-Binding Proteins ; Endothelium, Vascular ; drug effects ; metabolism ; Female ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; metabolism ; physiology ; Humans ; Milk Proteins ; Phosphorylation ; Pregnancy ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; STAT3 Transcription Factor ; STAT5 Transcription Factor ; Signal Transduction ; Trans-Activators ; metabolism ; Transcription, Genetic ; Tyrosine ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVESTo investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.

METHODSOne hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.

RESULTS4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).

CONCLUSIONSIn ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.

Adaptor Proteins, Signal Transducing ; Base Pair Mismatch ; Carrier Proteins ; Chromosomal Instability ; Cystadenocarcinoma, Mucinous ; genetics ; DNA Methylation ; DNA Repair ; DNA, Neoplasm ; genetics ; DNA, Satellite ; Female ; Genes, Neoplasm ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics