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.

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

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.

To investigate the effect of CDR1/CDR2 or MDR1 genes overexpression on oxidative stress in Candida albicans,we evaluated the effect of H2O2 on cell viability in C.albicans overexpressing genes CDR1 /CDR2 or MDR1 and their parent strains.After establishing an oxidative stress model with H2O2,we detected reactive oxygen species (ROS),mitochondrial membrane potential (Δψm) and the expression of oxidative stress response-related genes (CAP1 and GRP2) and ROS clearance related-genes (SOD2 and SOD5).The results showed that C.albicans growth were inhibited by 100％ after the treatment of 5 mmol/L H2O2.HeO2 caused more ROS accumulation and Δψm reduction in parent strains than in CDR1/CDR2 or MDR1 genes overexpressed strains (P＜0.05).Compared to parent strains,the up-regulated expression of CAP1 and GRP2 were relatively less in CDR1/CDR2 or MDR1 genes overexpressed strains,moreover,the down-regulated expression of SOD2 and SOD5 were also relatively less in CDR1/CDR2 or MDR1 genes overexpressed strains (P＜0.05).In conclusion,the overexpression of CDR1/CDR2 and MDR1 genes could reduce the oxidative stress response and enhance the adaptability of C.albicans to oxidative stress.

OBJECTIVETo observe the effect of Chinese herbal extract HuNan A-1 (HNA-1) on the thymic output function in Simian immunodeficiency virus (SIV) chronically infected rhesus macaques.

METHODSEight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months, and then they were randomly divided into the treatment group and the control group, 4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage, once daily for 2 successive months, while those in the control group were administered with equal volume of normal saline by gastrogavage, once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts, as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles (TREC) were detected using real-time fluorescent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue, CD4 counts, naive CD counts, and TREC were analyzed.

RESULTSThere was no statistical difference in body weight, viral loads, absolute CD ratios between the two groups after treatment (P > 0.05). The altered TREC multiple showed an obvious decreasing tendency in the control group, while it showed an increasing tendency in the treatment group (P < 0.05). In both groups, destroyed structures of the thymus tissue could be seen, filled with pink unstructured material. Increased connective tissues, lowered connective cell density, and confused arrangement could also be seen in the two groups, with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum (r = 0.926, P = 0.001). Naive CD4 counts were positively correlated with CD4 counts (r = 0.961, P = 0.005).

CONCLUSIONSTREC content determination, as a marker of newly thymic emigrants, could be taken as a testing method for evaluating the thymic output function. Besides, HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents, naive CD4 counts, and pathologies of thymus tissues, especially in late infection stage.

Animals ; CD4 Lymphocyte Count ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Macaca mulatta ; Plant Extracts ; pharmacology ; Random Allocation ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; Simian Immunodeficiency Virus ; Thymus Gland ; drug effects ; Viral Load

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

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.

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.