Human astrocyte cells were used as a normal control A total of 4

Human astrocyte cells were used as a normal control. A total of 47 paraffin-embedded primary tumors and 11 normal brain tissue (internal decompression in cerebral trauma) samples and used for semiquantitative reverse transcription-PCR and immunostaining had been obtained from 58 patients (30 female and 28 male patients; median age of 45.5 with a range of 11 to 74 years) undergoing curative surgery at the First Affiliated Hospital of Soochow University (Suzhou, China). A total of 26 tumor biopsy specimens and 7 corresponding normal brain tissue samples stored in liquid nitrogen (14 female and 19 male patients; median age of 47.4

with a range of 13 to Elacridar molecular weight 79 years) had also been obtained earlier from patients undergoing curative surgery at 3-deazaneplanocin A datasheet the First Affiliated Hospital of Soochow University (Suzhou, China) with informed consent. Clinical stage was judged according to the 2007 WHO classification of tumors of the central nervous

system [16]. The use of all clinical materials in this study was approved by individual institutional Ethical Committees. Serum and cerebrospinal fluid samples Serum samples were obtained with written informed consent from 8 healthy individuals and from 12 spongioblastomas, 6 low-grade gliomas, and 20 benign tumor patients in their neuronal system, i.e. the pituitary tumor, meningioma, nerve sheath tumor, and acoustic nerve tumor. The median age of these samples (20 males and 26 females) was 50.1 with a range of 26 to 79 years. Cerebral fluid samples from a total of 36 cancer patients and 6 healthy control individuals were also selected with informed consent from 26 males and 16 females (median Cobimetinib in vitro age of 48.9 with a range of 26 to 79 years). These 36 cancer cases included 14 spongioblastomas, 11 low-grade gliomas, and 11 patients with benign tumor in the neuronal system (pituitary tumor, meningioma,

nerve sheath tumor, acoustic nerve tumor, etc.). The serum and cerebrospinal fluid samples in this study were obtained at the time of diagnosis, centrifuged, and the supernatants were stored in liquid nitrogen. RNA preparation and cDNA synthesis Total cellular RNAs from cell lines and tissues were extracted and purified by using the Trizol reagent (Invitrogen, Inc.) according to the protocol of the supplier. Before RNA extraction, individual tissue samples were preexamined by frozen section histologic examination to document the histopathologic appearance of the selleck chemicals llc specimen. About 10 μg total RNA from each sample was reversely transcribed to single-stranded cDNAs using random hexamers (Shanghai Sangon, Inc.) as primer and M-MLV reverse transcriptase (Promega, Inc.).

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