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

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.

10.3969/j.issn.1000-8179.2013.12.005

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

A transmissible gastroenteritis virus strain was isolated from suspect samples in Sichuan province and identified by ST cell culture, direct fluorescent antibody test (FA), neutralization test (NT), TME examination and some other methods, then it was named SC-Y. The isolated strain could produce obvious cytopathic effects (CPE), The TCID50 was 10(-3.664)/0.05 mL, The neutralization index is 52.5. cDNA fragments covering the complete genome were amplified by the long reverse transcription PCR. The amplified fragments were further cloned and sequenced. The genome of SC-Y strain was assembled by BioEdit. The length of complete genome was 28590 nucletides, and was composed of 7 ORFs, which was flanked by untranslated regions (UTRs) with 315 bases at the 5'-end and 277 bases at the 3'-end. Phylogenetic analysis based on genome suggested that SC-Y might belong to same subgroup with Purdue strain.
Animals ; Base Sequence ; DNA, Viral ; chemistry ; Fluorescent Antibody Technique, Direct ; Microscopy, Electron ; Neutralization Tests ; Phylogeny ; Swine ; Transmissible gastroenteritis virus ; classification ; genetics ; isolation & purification ; ultrastructure

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.

Objective To evaluate the clinical efficacy and safety of domestic Sirolimus-eluting stents (Firebird) and imported Zotalimus-eluting stents ( Resolute) in the treatment of patients with unprotected left main coronary artery disease ( ULMCA) . Methods We retrospectively enrolled 76 patients with ULMCA treated by percutaneous coronary intervention (PCI) under the guidance of IVUS in our hospital. According to the different stents used in the procedure, the patients were divided into two groups: Domestic Sirolimus-Eluting Stents group (Firebird group, n = 42) and Imported Zotarolimus-Eluting Stents group (Resolute group, n = 34) . We analyzed the baseline characteristics, coronary artery lesion characteristics, stenting strategies and any changes in left ventricular ejection fraction ( LVEF) in both groups and investigated the long-term clinical outcomes. Results There were no significant differences in the baseline characteristics, the SYNTAX scores of the coronary artery lesion and the rate of complete revascularization between the two groups. Compared with that in Firebird group, there were more cases involving the distal left main (79. 4% vs. 45. 2% , P ﹤ 0. 05) and more patients using two stents strategies (29. 4% vs. 7. 1% , P ﹤ 0. 05) in the Resolute group. The change in LVEF post-PCI had no difference between the two groups. The patients were followed up for a mean of (23. 3 ± 10. 7) months. During the follow-up period, the occurrence of MACCE had no significant difference between the two groups. In the Firebird group, there were one sudden cardiac death, one nonfatal myocardial infarction, one stroke and five patients with recurrence of angina pectoris. In the Resolute group there were one sudden cardiac death, one target lesion revascularization and four patients with recurrence of angina pectoris. Conclusions Compared with the imported Zotalimus-Eluting Stents, the domestic Sirolimus-Eluting Stents are safe and effective in the treatment of patients with unprotected left main lesions under the guidance of IVUS. The two kinds of stent showed similar long-term clinical outcomes.

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.