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.

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 establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.

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.

BACKGROUND:Travoprost can increase human ciliary muscle cell interspace, decrease uveoscleral outflow resistance and then decrease intraocular pressure. But whether this action pathway is conducted by enhancing the expression of matrix metalloproteinase (MMP) in the ciliary muscle cells remains unclear.OBJECTIVE:To observe the effect of travoprost on the expression of MMP-2 in the human ciliary muscle cells.DESIGN:Controlled observation analysis.SETTING:Zhongshan Ophthalmic Center,Sun Yat-sen University.MATERIALS:This study was Carried out in the Zhongshan Ophthalmic Center,Sun Yat-sen University between August 2005 and April 2006.Donor was from the unilateral eyeball of a youth patient,who was dead within one hour,had no any disease (informed consent was obtained from the relatives) in the Zhongshan Ophthalmic Center, Sun Yat-sen University.Rabbit anti.human MMP-2 polyclonal antibody (Boster Bioengineering Co.,Ltd,Wuhan),and travoprost (86610F,0.004% solution,ALCON company.USA) were used in this study.METHODS: Experimental intervention: 1μmol/L travoprost was added into bovine serum-free medium of human ciliary muscle cell, serving as experimental group,and meanwhile,the cells which were not interfered by drugs were taken as control group.In the experimental group,cells were harvested 6, 12,and 24 hours after travoprost being added.Experimental evaluation:MMP-2 gene and protein expressions in the human ciliary muscle cells in each group were detected by RT-PCR and ELISA methods.The activity of MMP-2 in each group was detected by Zymography technique.MAIN OUTCOME MEASURES:MMP-2 mRNA expression in the human ciliary muscle cell,MMP-2 protein expression and MMP-2 activity in the extracellular fluid.RESULTS:①In the experimental group, at 6,12 and 24 hours after travoprost being added,the relative expression of MMP-2 mRNA was gradually increased (F=236.959,P＜0.01).②In the experimental group,at 6,12 and 24 hours after travoprost being added,MMP-2 protein was also gradually increased with time (F=38.110,P＜0.01).③Zymography technique detection showed that in the experimental group,at 6,12 and 24 hours after travoprost adding,MMP-2 activity was gradually enhanced with time (F=74.348,P＜0.01).CONCLUSION:After human ciliary muscle cell cultured in vitro being subjected to the intervention of travoprost.MMP-2 expression is gradually increased with action time of travoprost, and meanwhile MMP-2 activity is also gradually enhanced.

BACKGROUND: Cristata L flavonoid is a kind of plant estrin, which possesses multiple physiological function, has no toxicity and adverse effect, and is effective in treating and preventing osteoporosis.OBJECTIVE: To study the effect of cristata L flavonoid on bone morphogenetic protein, urine inorganic salt and content of lysozyme of rats with diabetes mellitus (DM).DESIGN: Randomized grouping design and controlled study.SETTING: Pharmaceutical Laboratory of Xinxiang Medical College.MATERIALS: The experiment was completed in the Xinxiang Medical College from September to December 2003. Totally 24 health male SD rats were randomly divided into three groups: normal control group, diabetes mellitus (DM) group, DM + cristata L flavonoid group with 8 in each group.were injected intraperineally with 60 mg/kg streptozotocin, and 48-72 hours later, blood of rear caudal vein was collected to measure total blood glucose.Rats were determined as DM if the blood glucose was ≥ 16.7 mmol/L; oth erwise, 60 mg/kg streptozotocin was injected once more. After modeling,cristata L flavonoid and rats in normal control group were given the same was measured with atomic absorbency method and content of urine sodium histochemistry of bone morphogenic protein-2 (BMP2) in bone was detected with streptavidin tagged by peroxidase and immunohistochemic expression lysozyme reflected reabsorption function of renal tubule was measured with with t-test. MAIN OUTCOME MEASURES: Comparison between content of calcium, sodium, kalium in urine and lysozyme and immunohistochemic expression of BMP2 10-week intervention later.calcium and sodium in urine in DM model group were obviously higher than those in normal control group, but content of kalium in urine was obviously lower (P ＜ 0.05-0.01). Contents of calcium in urine in DM +cristata L flavonoid group were obvioualy lower than those in DM model group and normal control group were obviously higher than those in DM model group, and content of lysozyme in urine in those groups was obviously lower than that in DM model group (P ＜ 0.05-0.01).CONCLUSION: Expression of BMP2 of DM rats is decreased, but after supplement of cristata L flavonoid compound, the expression is increased.In addition, output of urine calcium and sodium in DM rats is decreased,and reabsorption function of renal tubule is increased.

To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; growth & development ; physiology ; ultrastructure ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Humans ; Virus Release ; Virus Replication

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