Tumour-suppressive microRNA-144-5p directly targets CCNE1/2 as potential prognostic markers in bladder cancer.

BACKGROUND: Analysis of a microRNA (miRNA) expression signature of bladder cancer (BC) by deep-sequencing revealed that clustered miRNAs microRNA (miR)-451a, miR-144-3p, and miR-144-5p were significantly downregulated in BC tissues. We hypothesised that these miRNAs function as tumour suppressors in BC. The aim of this study was to investigate the functional roles of these miRNAs and their modulation of cancer networks in BC cells. METHODS: The functional studies of BC cells were performed using transfection of mature miRNAs. Genome-wide gene expression analysis, in silico analysis, and dual-luciferase reporter assays were applied to identify miRNA targets. The association between miR-144-5p levels and expression of the target genes was determined, and overall patient survival as a function of target gene expression was estimated by the Kaplan-Meier method. RESULTS: Gain-of-function studies showed that miR-144-5p significantly inhibited cell proliferation by BC cells. Four cell cycle-related genes (CCNE1, CCNE2, CDC25A, and PKMYT1) were identified as direct targets of miR-144-5p. The patients with high CCNE1 or CCNE2 expression had lower overall survival probabilities than those with low expression (P=0.025 and P=0.032). CONCLUSION: miR-144-5p functions as tumour suppressor in BC cells. CCNE1 and CCNE2 were directly regulated by miR-144-5p and might be good prognostic markers for survival of BC patients.

[Pharmacokinetics and clinical studies of aztreonam in neonates and premature infants].

Pharmacokinetics and clinical evaluation of aztreonam (AZT) in the neonates and premature infants were studied with the following results: 1. Serum concentrations of AZT in 60 minutes intravenous drip infusion of AZT 20 mg/kg to 6 cases of neonates with 2 to 22 days of age were 45.5 +/- 0.87 micrograms/ml immediately after the completion of intravenous drip infusion, 37.8 +/- 1.62 micrograms/ml 1 hour after, 31.2 +/- 1.92 micrograms/ml 2 hours after and 19.7 +/- 2.36 micrograms/ml 4 hours after, respectively. Serum half-life was 3.61 +/- 0.53 hours on the average. 2. Urinary excretion rate 6 hours after intravenous drip infusion was 26.4 +/- 6.86% on the average. 3. Clinical evaluation was given to 1 sepsis case of 7-days of age and it was effective. There was no abnormal clinical or laboratory finding considered to be associated with AZT.

[Cord blood stem cell transplantation for a patient with acute myelogenous leukemia (M1)].

A five-year-old boy with acute myelogenous leukemia in relapse was treated by HLA-matched cord blood stem cell transplantation. The patient was preconditioned with 16 mg/kg of busulfan, 15 mg/kg of thiotepa and 90 mg/kg of cyclophosphamide and 2.45 x 10(7)/kg of cord blood mononuclear cells were infused to the patient on October 19th 1995 without the prophylaxis of graft-versus-host disease (GVHD). From the fifth day following the transplant, rG-CSF was administered at a dose of 300 micrograms/m2/day. Hematopoietic recovery was obtained as following; WBC over 1000/microliters was on +18 day, neutrophil over 500/microliters was on +20 day, reticulocyte over 20/1000 was on +28 day and platelet over 50 x 10(2) microliters was on +91 day. Engraftment was confirmed by DNA restriction fragment length polymorphism (VNTR) on +28 day. In spite of absence of prophylaxis of GVHD, the patient did not develop any signs of GVHD, and leukemia relapsed on +105 day. The patient died of leukemia relapse on +251 day. This is the first case of cord blood stem cell transplantation in Japan.