Aged ; Aged, 80 and over ; Biomarkers, Tumor ; urine ; Carcinoma, Transitional Cell ; urine ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; RNA, Messenger ; urine ; Reverse Transcriptase Polymerase Chain Reaction ; Urinary Bladder Neoplasms ; urine

Background and Objective:Studies on survivin over the past 5 years have shown that survivin participates in the genesis of several human cancers,including bladder cancer.Recent studies have indicated that survivin splice variants appeared to have unique subcellular localizations and functions as well.This study was to explore the roles of survivin and its two splice variants survivin-2B and survivin-△Ex3 in transitional cell carcinoma of bladder (BTCC).Methods:The relative amount of survivin,survivin-2B, and survivin-△Ex3 mRNA of fresh carcinoma tissues from 60 patients with BTCC and 12 non-cancerous bladder tissues were detected by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR),and the relationships of their expression levels in different pathologic grades to clinical stages of bladder cancer were analyzed.The time of follow-up was 4-24 months.Results:Survivin,survivin-2B,and survivin-△Ex3 mRNA were detected in all BTCC tissues,and their relative expressions were 0.333±0.163,0.056±0.017, and 0.124±0.096,respectively. In the control group,three and four samples expressed survivin and survivin-△Ex3 mRNA respectively,and all samples expressed survivin-2B mRNA.The expressions of survivin and survivin-△Ex3 mRNA were positively correlated with the pathologic grades and clinical stages(0＜r'_s＜1,P＜0.05),however,survivin-2B mRNA was negativly correlated with those (-1＜r'_s＜0,P＜0.05).Conclusion:Detecting the expression levels of survivin and its two splice variants survivin-2B and survivin-△Ex3 mRNA in BTCC by real-time PCR could have potential values to evaluate tumor progression and recurrence rate.

BACKGROUNDBladder cancer is the most common type of urinary system tumours. It is frequently associated with genetic mutations that deregulate the cell cycle and render these tumours resistant to apoptosis. Survivin, a newly discovered member inhibitor of apoptosis protein (IAP) family in several human cancers, by inducing cell proliferation and inhibiting apoptosis is frequently activated in bladder cancer. We studied the influence of small interfering RNA (siRNA) targeting survivin on the biological behaviour of bladder cancer cells.

METHODSA double strand survivin target sequence specific siRNA was designed and synthesized. After transfection of bladder cancer cell line T24 by siRNA/liposome complex with increasing concentrations (50200 nmol/L), the transfectant cells were intratumourally injected at different doses (5 microg or 50 microg). The effects were measured in vitro and in vivo.

RESULTSThe selected siRNA efficiently down-regulated survivin mRNA expression in a dose and time dependent manner. The maximal effect was achieved at the concentration of 100 nmol/L, at which survivin expression level was down-regulated by 75.91%. The inhibition rate of cell growth was 55.29% (P < 0.01) and the markedly increased apoptotic rate was 45.70% (P < 0.01). In vivo intratumoural injection of 50 microg siRNA-survivin could notably prevent the growth of bladder cancer (P < 0.01) in xenografted animals.

CONCLUSIONThe application of siRNA-survivin could markedly inhibit survivin expression in bladder cancer cell line by inducing apoptosis and inhibiting the growth of the tumour. It may become a new gene therapy tool for bladder cancer.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Mice ; Mice, Inbred BALB C ; Microtubule-Associated Proteins ; analysis ; antagonists & inhibitors ; genetics ; Neoplasm Proteins ; analysis ; antagonists & inhibitors ; genetics ; Neoplasm Transplantation ; RNA, Small Interfering ; pharmacology ; therapeutic use ; Transfection ; Urinary Bladder Neoplasms ; pathology ; therapy

OBJECTIVETo investigate spike wave reduction in electrocorticography (EcoG) monitoring for evaluating the outcomes of epilepsy surgery.

METHODSThe epileptogenesis lesions in the target cortex was localized accurately using an EcoG monitoring system in 20 surgical patients with intractable EP. The spike numbers within 60 s were recorded before and after surgical resection of the epileptogenic focus. In cases where the spike number within 60 s was reduced by over 80% after the resection, the surgery was terminated, otherwise extended lesion resection, corpus callosotomy or multiple subpial transection (MST) was carried out with ECoG monitoring, and the spike number within 60 s was recorded. Antiepileptic drugs were routinely prescribed after the operations.

RESULTSTwelve patients exhibited a spike wave reduction by over 80% after resection or extended resection of the lesions, including 4 with cavernomas in the nonfunctional area, who showed a spike wave reduction by over 80% after extended resection of the cortex around the tumor. The reduction was still less than 80% in 4 patients with hippocampal sclerosis and 3 with neurogliocytoma in the functional area after the operations. According to the Engel assessments, 13 cases were in level I, 3 cases in level II, 1 in level III, and 3 in level IV. Seventeen patients responded favorably to the treatment, with a total effective rate of 85%.

CONCLUSIONFor extra-temporal lobe epilepsy, a postoperative spike wave reduction beyond 80% indicate favorable outcome of the surgery, otherwise poor prognosis is expected. But in cases of temporal lobe epilepsy, no direct association is found between spike wave reduction and the prognosis of the patients.

Adolescent ; Adult ; Brain Mapping ; Cerebral Cortex ; physiopathology ; Child ; Child, Preschool ; Electroencephalography ; Epilepsy ; physiopathology ; surgery ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; Treatment Outcome ; Young Adult

Objective To describe the survival state and to investigate the risk factors of death on patients with subarachnoid hemorrhage (SAH). Methods Age, past history, number of encephalic region suffering SAH, laboratory examination indexes, therapeutic measures, complications and prognosis of 174 patients with SAH were followed-up and investigated. The survival states and risk factors of death of the patients with SAH were identified by both Kaplan-Meicr survival analysis and Cox proportional risk model. Results There were 10 patients (5.75%) losing follow-up investigation and 164 patients with SAH completed the follow-up investigation. 66 patients died and the longest follow-up invcstigation time was 5.64 years. The survival rates of 28 days, 1 year and 3-5 years were 70.60%,63.40% and 57.20% respectively. The treatment of nimotop, aneurysm occlusion treatment and aneurysm embolotherapy could decrease the death of SAH. At the same time, advanced age, the long time smoking, hyponatremia, the rising of leucocyte in acute stage, repeated hemorrhage and cerebral angio spasm were the independent risk factors to the death of patients. Conclusion Prognosis of patients with advanced age, the rising of leucocyte in acute stage, gastrointestinal blooding, hyponatremia, repeated hemorrhage and cerebral angio spasm were unfavorable. When giving patients with aneurysm, the aneurysm occlusion and embolotherapy and nimotop treatment, the death risk could be reduced.

OBJECTIVETo explore the molecular mechanisms involved in the patient with congenital FV deficiency.

METHODSActivity of FV was determined by biochemical method. The PCR products of FV gene was analysed by directly sequencing or sequencing after cloned into T-vector. The mutative FV gene was analysed by restriction enzyme analysis in the proband and her family members.

RESULTSA homozygous missense mutation G5729T resulting in Gly1880Val was revealed in the proband and confirmed in the family screening. Structure-function studies of the factor V mutants (Gly1880Val) demonstrated the importance of Gly1880 for structural stability of the Factor V.

CONCLUSIONG5729T mutation of FV gene is related to the pathogenesis of congenital FV deficiency.

Adult ; DNA Mutational Analysis ; Factor V ; genetics ; metabolism ; Factor V Deficiency ; blood ; congenital ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Polymerase Chain Reaction

OBJECTIVETo study the influence of small interfering RNA (siRNA) targeting survivin on the biological behavior of bladder cancer.

METHODSOne pair of survivin target sequence-specific siRNA was designed, then siRNA/liposome complex was used to transfect bladder cancer cell line-T24. The efficiency of transfection and the apoptosis were detected by flow cytometry. The transcriptional level of survivin was analyzed using real time PCR. DNA sequence corresponding to siRNA targeting survivin was cloned into pRNAT-U6.1/Neo to produce plasmid targeting surviving.

RESULTSThe ratio of T24 cells releasing fluorescence in total cells were 92.3%; siRNA-survivin efficiently down-regulated survivin expression (mRNA) in a dose-and time-dependent manner. Its maximal effect was achieved at the concentration of 100 nmol/L, at which survivin expression level was down regulated by 75.91%. Similar results were found in the inhibition ratio of cell growth, which was 55.29%(P< 0.01). Simultaneously the apoptotic rate was markedly increased, which was 45.70%(P< 0.01). After cutting the vector with Bam H I and Hin d III and ligating the vector with the insert by using T4 ligase, the recombinant vector was confirmed by restriction digestion and DNA sequencing.

CONCLUSIONThe application of siRNA-survivin can markedly inhibit survivin expression in bladder cancer cell line, induce apoptosis and inhibit the growth of the tumor. It may be a new gene therapy tool for bladder cancer. The successful construction of the siRNA expressing plasmid will facilitate the application of RNA interference technique, and lay a foundation for further studies on the function of surviving.

Cell Line, Tumor ; Genetic Vectors ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Urinary Bladder Neoplasms ; genetics ; pathology

OBJECTIVETo investigate the effects of the downregulated expression of the prostate androgen regulated (PAR) gene on the cell cycle and apoptosis of PC3 cells as well as on the expression level of Bcl-2/Bax.

METHODSAfter transfecting PC3 cells with small interfering RNA (siRNA) targeting PAR, we detected the inhibitory effect of PAR depletion on the proliferation of the PC3 cells by MTT assay, determined their apoptosis by flow cytometry, and measured the expression levels of Bcl-2 and Bax by Western blot.

RESULTSThe expression of PAR was suppressed by siRNA, the G2-M phase PC3 cells were increased to (29.95 +/- 3.25)%, and the apoptosis of the cells was enhanced to (20.61 +/- 2.73)%, with statistically significant difference from the control group (P < 0.01). Western blot showed a decreased expression of Bcl-2, an increased expression of Bax, and an elevated ratio of Bax to Bcl-2.

CONCLUSIONDownregulation of the PAR expression increases the Bax/Bcl-2 ratio and Bax expression, and thus induces the G2-M phase arrest and apoptosis of PC3 cells.

Apoptosis ; Cell Cycle Checkpoints ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Prostate ; metabolism ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the expressions of endothelial nitric oxide synthase (eNOS) and connexin 43 (Cx43) in the penile tissue of rats with diabetes mellitus induced erectile dysfunction (DMED) and their correlation with DMED.

METHODSSD rat models of DM were established by intraperitoneal injection of alloxan, and 8 weeks later, apomorphine was administered to induce ED in the DM models. The expressions of eNOS and Cx43 were measured by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively.

RESULTSAlloxan did not influence the expressions of eNOS mRNA and Cx43 mRNA in the penile tissue. Compared with the DM models, the expression of eNOS mRNA significantly decreased in the DMED group (0.155 +/- 0.157 vs 0.508 +/- 0.242, P < 0.01), while that of Cx43 mRNA markedly increased (0.993 +/- 0.157 vs 0.545 +/- 0.138, P < 0.01), with a negative correlation between the two expressions (r = -0.987). The same results were shown by immunohistochemistry in the penile smooth muscle cells of the DMED rats.

CONCLUSIONThe decrease of eNOS expression in the penile tissue might play a key role in the development of ED in diabetic patients, while the accompanying compensative elevation of the Cx43 level has yet to be further studied.

Animals ; Connexin 43 ; biosynthesis ; genetics ; Diabetes Mellitus, Experimental ; complications ; Erectile Dysfunction ; etiology ; physiopathology ; Immunohistochemistry ; Male ; Nitric Oxide Synthase Type III ; biosynthesis ; genetics ; Penis ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the enhancing effect of all-trans retinoic acid (ATRA) on the bystander effect of the herpes simplex virus thymidine kinase(HSV-TK)/ganciclovir (GCV) against androgen unresponsive prostate cancer.

METHODSThe bystander effect of the HSV-TK/GCV system was measured by methyl thiazolyl tetrazolium (MTT) assay on PC-3 cells before and after ATRA treatment. The growth and the histopathology of transplant tumors were observed in 4 groups of nude mice with prostate cancer.

RESULTSATRA augmented significantly the bystander effect of the HSV-TK/GCV system by reducing TK positive PC-3 cells from 50% to 30% (P < 0.05). HSV-TK showed an inhibiting effect, while ATRA with the HSV-TK/GCV system produced significant effect on prostate cancer 1 week earlier than the former (P < 0.05).

CONCLUSIONATRA can argument the in vivo and in vitro bystander effect of the HSV-TK/GCV system in the treatment of androgen unresponsive prostate cancer.

Animals ; Antineoplastic Agents ; pharmacology ; Bystander Effect ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; methods ; Humans ; Male ; Mice ; Mice, Nude ; Prostatic Neoplasms ; genetics ; pathology ; therapy ; Reverse Transcriptase Polymerase Chain Reaction ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; metabolism ; Tretinoin ; pharmacology ; Xenograft Model Antitumor Assays ; methods