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

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 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

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

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 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

The aim of this study was to investigate the role of mesenchymal stem cells in the hematopoietic reconstitution of patients who had received haploidentical allogeneic hematopoietic stem cell transplantation (hi-allo-HSCT). 15 patients who underwent treatment with both MSCs and HSCs, were selected as study group, while 20 patients receiving only HSCT were taken as control. Bone marrow samples were obtained from iliac crest aspirates at several times after HSCT for the isolation, purification and expansion of MSCs. The confluent ratio and time were measured and compared with those of the control. The peripheral blood samples were obtained from patients, then absolute neutrophil and platelet counts were assayed. From day 4 before transplantation to day 28 after transplantation, serum was obtained every four days from patients of the two groups, and then 3 cytokines as SDF-1alpha, TPO and IL-11 were detected by ELISA. The results indicated that as compared with the control group, the ratio of primary confluent layer formation of MSCs in study group was obviously higher (27.3%) (p < 0.01), and the confluence time in culture was significantly less (p < 0.05). In the study group, the concentration of SDF-1alpha amounted to peak value (2975.19 +/- 681.56 pg/ml) on the 8th day after HSCT, which was obviously higher than that before HSCT (2403.70 +/- 522.39 pg/ml, p < 0.05), whereas in the control, the concentration of highest point of SDF-1alpha reached to peak valve (2280.60 +/- 701.25 pg/ml) on the 16th day after HSCT, which was less than that before HSCT (2701.46 +/- 483.21 pg/ml, p < 0.05). The concentration of TPO and IL-11 was higher in study group compared with the control from day 16 to 28 after HSCT (p < 0.05). It is concluded that the transfusion of MSCs combined with hi-all-HSCT may improve the injured state of the hematopoietic microenvironment in bone marrow of patients during allo-HSCT.
Adolescent ; Adult ; Bone Marrow ; metabolism ; pathology ; Chemokine CXCL12 ; metabolism ; Child ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic System ; Humans ; Interleukin-11 ; metabolism ; Mesenchymal Stem Cell Transplantation ; methods ; Middle Aged ; Thrombopoietin ; metabolism ; Young Adult

OBJECTIVETo study the prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi.

METHODSA total of 7407 people with Zhuang nationality, aged 16 and over were surveyed. Questionnaire was administered to each subject under survey, while X-ray and serum rheumatoid factor were carried out to all those who gave positive answers. Physical examinations were done to the suspicious cases by experts on rheumatism.

RESULTSwere compared with those in 6826 people of Han ethnicity living in the same district.

RESULTSThe prevalence rates of rheumatoid arthritis in Zhuang nationality population was 0.27% when comparing to the Han population which was 0.28% (P > 0.05). Rates of awareness on rheumatoid arthritis in Zhuang and Han population were 5.0% and 10.5% (P > 0.05). After the diagnosis of rheumatoid arthritis was made and among patients who had received the treatment, the rates were 0% vs. 5.25%.

CONCLUSIONThe prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi was not significantly different than that in Han ethnic group. However, the rates on awareness and the treatment of rheumatoid arthritis were still under satisfaction.

Adult ; Aged ; Arthritis, Rheumatoid ; epidemiology ; Asian Continental Ancestry Group ; ethnology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence

BACKGROUNDHumoral immunity is an important factor for long-term survival of renal allograft. Here we performed a four-year follow-up to explore the clinical significance of monitoring anti-human leukocyte antigens (HLA) and anti-major histocompatibility complex class I-related chain A (MICA) antibody expression after kidney transplantation.

METHODSWe obtained serial serum samples from 84 kidney transplant patients over a four-year period. All patients were followed up at least 6 months after transplantation and had at least two follow-up points. Anti-HLA and anti-MICA antibody titres and serum creatinine (SCr) levels were evaluated at each follow-up. Patients were divided into 4 groups: HLA(+) MICA(-), HLA(-)MICA(+), HLA(+)MICA(+) and HLA(-)MICA(-). The impact of post-transplant antibody level on kidney allograft function was evaluated.

RESULTSAntibodies were detected in 38.1% (32/84) of the renal allograft recipients. HLA, MICA and HLA+MICA expression was observed in 18.89%, 14.44% and 5.93% of the recipients respectively. The most frequent anti-HLA and anti-MICA specific antibodies identified were A11, A24, A29, A32, A33, A80; B7, B13, B37; DR17, DR12, DR18, DR52, DR53, DR1, DR4, DR9, DR51; DQ7, DQ4, DQ8, DQ2, DQ9, DQ5, DQ6 and MICA02, MICA18, MICA19, MICA07, MICA27. As the time after transplantation elapsed, more recipients developed de novo antibody expression. Total 11.91% (10/84) of the recipients had de novo antibody expression during the follow up. The average level of SCr and the percentage of recipients with abnormal allograft function were significantly higher in recipients with anti-HLA and/or anti-MICA antibody expression than those without. The appearance of anti-HLA and anti-MICA antibody expression always preceded the increase in SCr value.

CONCLUSIONSAnti-HLA and anti-MICA antibody expression has predictive value for early and late allograft dysfunction. The presence of donor specific antibody is detrimental to graft function and graft survival.

Female ; Follow-Up Studies ; Graft Survival ; HLA Antigens ; immunology ; Histocompatibility Antigens Class I ; immunology ; Humans ; Isoantibodies ; analysis ; Kidney Transplantation ; Male ; Minor Histocompatibility Antigens