Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Gene Expression Regulation ; drug effects ; Humans ; NF-kappa B ; metabolism ; Scorpion Venoms ; pharmacology

10.3969/j.issn.1000-8179.2013.12.005

Objective To observe the effect of systemic chemotherapy on conditions of tumor infiltrating,metastasis and disease-specific survival (DSS) for advanced retinoblastoma (RB).Methods Forty-one patients with advanced RB who received enucleation were enrolled in this study.There were 26 males and 15 females,age at diagnosis was ranged from 2 to 72 months,with a mean of 23.08 months.There were 16 bilateral patients and 25 unilateral patients;13 group D eyes and 28 group E eyes.16 patients received enucleation as the primary treatment (operation group),25 eyes received chemotherapy before enucleation (chemotherapy group).There was no significant statistical difference between two groups for the gender,unilateral and bilateral,international staging or diagnostic age (P＞0.05).The histopathology report was performed to assess the risk of postoperative tumor-node-metastasis staging (pTNM) in each patient,and the extent of tumor invasion in the optic nerve,choroid and anterior chamber was divided into 3 levels of low risk,medium risk and high risk.Five deaths were all in the group E with chemotherapy before enucleation.Using R software survival analysis software package survfit function,the application of Kaplan-Meier estimation method,DSS of RB children was calculated from the time of diagnosis,up to the date of the death of patient.DSS differences between chemotherapy,operation group and eye removal time (more than 3 months,less than 3 months) in group E RB children were analyzed.Results The proportion of high risk pTNM stage in chemotherapy group was significantly lower than the operation group.But there was no significant difference between the two groups in the overall risk classification (x2 =3.130,P=0.077).For group D eyes,the overall risk classification in chemotherapy group was significantly lower than the operation group (x2 =5.870,P=0.015).There was no significant difference between the two groups in the overall risk of group E eyes (x2 =0.020,P=0.889).The DSS in chemotherapy group and operation group were 0.71 and 1.00,respectively;the difference was significant (x2 =3.700,P=0.05).The DSS in children whose enucleation delayed for more than 3 months and children whose enucleation performed within 3 months were 0.64 and 1.00,respectively;the difference was significant (x2 =4.800,P=0.028).Conclusion Systemic chemotherapy did not reduce the risk of tumor invasion and metastasis in patients with advanced RB.Instead,it will reduce the DSS in group E eyes of RB.

Acute Disease ; Humans ; Lumbar Vertebrae ; Male ; Middle Aged ; Osteochondroma ; diagnosis ; pathology ; therapy ; Spinal Neoplasms ; diagnosis ; pathology ; therapy

Objective To investigate the relation among the results of thyroid fine needle aspiration cytology(FNAC),thyroid ultrasonography(US)and histopathologic diagnosis about the thyroid nodules detected by physical examination,meanwhile to analyze the etiopathogenesis of the nodtries and to evaluate the risk of thyroid cancer and the clinical diagnostic value of FNAC.Methods The data of thyroid FNAC results of the thyrroid nodules detected by physical examination in 271 cases were analyzed and compared with thyroid US and histopathologie diagnosis.Resuit (1)The FNAC results showed that the incidences of malignant and suspected mali-gnant lesions were 1.48%and 5.90%respectively.The rate of benign lesions was 78.60%and that of goiter was 29.15%,Hashimoto's thyroiditis 26.57%and thyroid adenoma 15.13%.Benign lesions were more common than malignant ones.(2)Comparison of the FNAC and US results of the thyroid nodules showed that of 96 cases with single nodule the rates of malignancy and suspected malignancy were 3.12%and 7.29%,but of 137 cases with multiple nodules the rates of the two lesions were 0.73%and 6.57%.In 108 cases with smaller nodules(≤1.5 cm)the rate of malignancy and suspected malignaney found with FNAC were 0.93%and 7.41%,while in 125 cases with greater nodules (＞1.5 cm)the rate of the two lesions were 2.40%and 6.42%.In 99 solid nodules the rates of malignancy and suspected malignancy were 2.02%and 12.12%,while in 85 cystic or mixed nodules the rates of the two lesions were 2.35%and 2.35%.In the above-mentioned three groups,only the suspected malignaney rate in solid nodules was higher than these in cystic or mixed ones with significant difference (P=0.013).(3)As compared with the cytological and histological diagnoses in 24 cases,the diagnostic accuracy of FNAC was 75.00%and the rates of false positive and false negative were 25.00%and 0,respectively.Conclusions The common causes of the thyroid nodules detected in physieal examination are goiter,Hashimoto's thyroiditis and thyroid adenoma.FNAC is a reliable method to define the benign or malignant nature of thyroid nodules with a high diagnostic accuracy.US features of the nodule alone,no matter it is single,solid or of greater size do not sufficiently increase the incidence of thyroid carcinoma.

Objective:To investigate the role of transcription factor SP1 decoy oligodeoxynucleotides(ODN) on expression of ? Gal in SV-40-PED cells.Methods:Immortalized porcine aortic endothelial cells of the PED line were cultured and transfected with ?1,3galactosyltransferase(?1,3GT) specific decoy ODN.Cells transfected with mismatch ODN was used as negative controls.Twenty-six hours later the cells were collected.The expression of ? Gal was determined with fluorescence microscope and Western blot.The expression of ?1,3GT mRNA was examined by RT-PCR.Results:Fluorescence microscopy observed the decreased fluorescence of ? Gal after decoy ODN transfection.Western blot showed that the average absorbance of the PED cells transfected with decoy ODNs was(48.2?0.9).It is 52.6% of the mock group(P0.05).Conclusion:?1,3GT gene reduce actually occurs following transfection of decoy ODN.Porcine endothelial cells can be the targets of decoy ODN.

Objective:To construct a short hairpin RNA(shRNA) eukaryotic expression vector of the survivin gene,and investigate its inhibitory effect on the survivin expression in human cervical cancer cells.Methods:We designed and synthesized 2 pairs of survivin-specific small interfering RNA primers(s1 and s2),cloned them into the eukaryotic expression vector pSilencer 2.1-U6 neo by DNA recombined techniques after annealing connection reaction,and transfected them respectively into human cervical cancer cell line HeLa using LipofectAMINE2000 after identificationby restrict endonuclease digestion and DNA sequencing.Then we selected the positive clones by G418 and detected the expression of survivin mRNA by semi-quantitative RT-PCR and that of the survivin protein by Western blot.Results:The survivin shRNA eukaryotic expression vectors pSilencer2.1-s1 and pSilencer2.1-s2 were successfully constructed.Positive clones were obtained by screening with G418 for 24 days,the survivin expression in HeLa cells decreased in different degrees after transfected with pSilencer2.1-s1 and pSilencer2.1-s2,and the latter showed a better interference efficiency.Conclusion:The shRNA eukaryotic expression vector of the survivin gene we constructed,with its improved interfering efficiency,has paved the way for further research on its role in regulating the biological behavior of cervical cancer cells.

Objective: To investigate the relationship between MALT1 mRNA expression in extranodal B cell lymphoma and the pathogenesis of MALToma and DLBCL, and to explore the effect of MALT1 mRNA expression on the clinicopatho-serve the expression of Bcl-2 and Ki-67 in 106 samples of extranodal B cell lymphoma.The mRNA of MALT1 was detected by RT-PCR.Clinicopathological data were reviewed.Follow-up and statistical analysis were performed.Results: Expression of MALT1 mRNA was statistically different between the cases with lymph node metastasis and those without lymph node mestastasis, and between staging Ⅰ-Ⅱ cases and stage Ⅲ-Ⅳ cases, The expression of MALT1 mRNA in cases with posi-tive expression of MALToma, DLBCL, and Bcl-2 was higher than in those without expression of the three indices (P<0.05).However, no significant difference was found in MALT1 mRNA expression between cases with Ki-67≥30% and those with Ki-67<30% (P>0.05).MALT1 mRNA-positive cases showed a worse survival status than MALT1 mRNA-negative cases (P<0.05).Conclusion: Expression of MALT1 gene is different between MALToma and DLBCL.Inhibition of apoptosis by Bcl-2 overexpression caused by activation of NF-κB may lead to the pathogenesis of MALToma and DLBCL.The expression of MALT1 mRNA is associated with the types of lymphoma, involvement of lymph node, stage of lymphoma, and patient sur-vival.The expression of MALT1 gene may be associated with poor prognosis of MALToma and DLBCL.

Objective To set a rapid,simple gene diagnosis method for Down syndrome.Methods Three short tandem repeats(D21S11,D21S1270,D21S1437)loci in or near Down syndrome critical region(DSCR) were analyzed and detected by polymerase chain reaction and DNA quantitative analysis in 11 core ancestry.Results There were four types by DNA quantitative analysis to different individuals at a short tandem repeats(STR) locus.In type one,a homozygote of one allelic gene was detected.In type two,a normal heterozygote of two allelic genes was found,the content or two DNA electrophoresis bands was approximately 1∶1.In type three,a Down syndrome patient of two allelic genes was discovered.The quantity of two electrophoresis bands was nearly 2∶1.In type four,the patient showed three DNA electrophoresis bands which the content was approximately 1∶1∶1.Conclusion A rapid gene diagnosis and prenatal diagnosis method for Down syndrome can be used for quantitative analysis of STR polymorphism loci.

AIM:To examine the difference of vascular remodeling between aorta and small artery in spontaneous hypertensive rats (SHR) and control rats.METHODS:Male SHR (20-week-old) were used as experiment group,and age matched male Wistar-Kyoto (WKY) rats were used as control group.The systolic blood pressure and body weight were measured once a week.At 43 weeks old,the rats were anaesthetized,blood samples were collected,and thoracic aorta and mesenteric small artery tissue were harvested.The morphological changes of the arterial tissue were observed with HE staining.The collagen and elastine fibers were detected by the Sirius red-Victoria blue staining.The protein expression of type Ⅰ and Ⅲ collagens were analyzed by confocal laser-scanning microscopy and Western blot.The changes of the vascular ultrastructure were imaged by transmission electron microscopy.The expression of proliferating cell nuclear antigen (PCNA) and the cell apoptosis in the arterial wall were examined by immunohistochemical method and TdT-mediated dUTP nick and labeling (TUNEL) detection.RESULTS:The inner diameter (ID) and luminal cross-sectional area (LCSA) of mesenteric small artery were decreased,whereas ratio of wall thickness (WT) to ID (WT/ID) and ratio of wall cross-sectional area (WCSA) to LCSA (WCSA/LCSA) were increased.Meanwhile,adventitia fibroblast migrated to the nedia,with overload collagens,especially collagen Ⅲ.Proliferation index (PI) and apoptotic index (AI) of the mesenteric small artery wall cells were increased.The ID,LCSA,WT/ID and WCSA/LCSA of the aorta were increased.Moreover,the vascular smooth muscle cells (VSMCs) showed hypertrophy and hyperplasia,with overload collagens.The PI and AI of the aortic wall cells were increased.CONCLUSION:The difference of vascular remodeling between the aorta and small artery is significant.The small artery mainly appears hyperplasia of matrix,especially the adventitial collagen Ⅲ.Meanwhile,the cell apoptosis in the small artery wall is increased.The aorta mainly appears hyperplasia and hypertrophy of media VSMCs.