Objective To screen and identify proteins that interact with receptor for advanced glycation end products (RAGE) with T7 select phage display system. Method GST-tagged fusion protein of RAGE was coated on glutathione suporflow resin and binding screens were performed with human lung T7 phage library. Results After 6 rounds of biopanning, 64 independent plaques were scraped and processed with EDTA. The inserted fragments in these plaques were amplified by PCR and the PCR products were purified by a gel recovery method. The sequences of the insertions were identified and analyzed with BLAST program in GeneBank. Twenty-five clones of encoding proteins were found. Conclusions T7 select phage display system is a convenient, rapid and effective method for screening binding proteins. This study provides an approach to explore complicated functions and novel mechanisms of RAGE-related signal transduction pathways.

Objective To investigate the clinical characteristics of adult Wilms'tumor in Chinese adults and to improve the recognition of adult Wilms'tumor.Methods The clinical data of 6 Wilms'tumor patients treated in our hospital were reviewed. Of them, there were 4 male patients and 2 female patients with mean age of 31 years old. Four patients with abdominal and flank pain and 2 patients with abdominal masses were the main clinical manifestations. One case accompanied with hematuria and another one had fever.Tumor masses were palpable by physical examination in 4 patients. Five patients had percussion tenderness over kidney region and 1 case had abdominal tenderness. Six cases underwent urinalysis and red blood cell was positive in 2 cases. There were no abnormal results in other laboratory tests. Chest X-ray was routinely performed before operation. Six patients underwent ultrasonography and all showed hypoechoic masses (5.0cm×5.0 cm-22.0 cm×25.0 cm) with clear margin. Four cases examined by CT and all had solid masses with inhomogeneous density and clear margin. On enhanced CT scan, 1 of 4 cases demonstrated irregular enhancement of the mass with local hemorrhagic necrosis area. The CT value ranged from 11-40 HU, with an average value of 22.5 HU, and increased to 35-78 HU, with an average value of 63.5 HU after administration of contrast. Two cases of MRI revealed solid mass defects.Three cases underwent IVU examination and showed the enlargement of renal umbra as well as thinning of renal calices caused by the compression of tumor and 1 case showed hydronephrosis.Thus, 5 cases were diagnosed with renal tumor and 1 case was diagnosed with cavitas pelvis tumor before operation. Five patients successfully underwent nephrectomies and 1 patient accepted oophorotomy with no subsequent complication.Retroperitoneal lymph node dissection(RPLND) was performed at the time of radical nephrectomy or oophorotomy in all patients. The tumor was found to be about 5-25 cm in size during operation. One of the tumors had invaded pancreas tail, one had invaded to the lateral abdominal wall and one had invaded into the inferior caval vein. Renal hilar lymph node was found to be exceeded 1.5 cm×1.5 cm in 2 patients. The post-operative incisal surface of the tumor presented with gray-yellow color. Capsule was intact in 4 patients. Three cases had hemorrhagic necrosis in tumor center. All the specimens sent for the pathological examination. None received adjuvant therapy before operation. Four patients received chemotherapy plus radiotherapy after surgery and other 2 patients received chemotherapy only. All patients were followed up with routine lab tests, ultrasonography and chest X-ray.Results Six patients were staged according to the National Wilms'Tumor Study staging system as follows: 1 case in stage Ⅰ, 2 cases in stageⅡ, 2 cases in stage Ⅲ and 1 case in stage Ⅳ. Nephrectomies or oophorotomy were performed in all patients successfully. Tumor diameters were from 4.5-25.0 cm, in average 11.8 cm.All cases had pathological diagnosis of Wilm's tumor. Five cases were classified into favorable histologic type and 1 case was in unfavorable histologic type. Two patients with lymph node metastasis and 3 patients with local invasion were observed. One patient with pulmonary metastasis was recorded. Six patients were followed up for 1-7 years. Five patients survived with no evidence of cancer recurrence. One patient died of pulmonary metastasis 2 years after surgery.Conclusions The majority of adult Wilms'tumors are found with local invasion and metastasis. The incidence of hematogenous metastasis in adult Wilms'tumor is higher than other types of renal tumor as well. The early diagnosis and radical surgical treatment and adjuvant chemoradiotherapy could improve the survival rate.

Objective To review the diagnosis and treatment of squamous cell carcinoma of renal pelvis. Methods The clinical data from October 1991 to May 2009 of eight cases of squamous cell carcinoma of renal pelvis were reviewed and analyzed retrospectively. The symptoms of the patients were hematuria (eight cases), pain (seven cases) and abdominal mass (one case). All patients underwent B-ultrasound and IVU examination and four cases underwent CT scan. Three cases were diagnosed as having a tumor before surgery. Five cases were diagnosed as renal calculus, two of the five cases were diagnosed by intraoperative frozen section. Radical nephroureterectomy were performed in four cases, nephrectomy in three cases and palliative resection in one case. Results Histological classification revealed that six cases were moderately differentiated, one case was well differentiated and one case was poorly differentiated. Two cases had stage pT1/pT2 and six cases had stage pT3/pT4. 2 cases had regional lymph nodes metastasis. Seven cases were followed-up. All patients died of tumor recurrence or metastasis. The median tumor specific survive time was six months (range from two months to 42 months). Conclusions Squamous cell carcinoma of renal pelvis is often occurs concurrently with urolithiasis which could lead to difficulty in diagnose before operation. As the most of the patients were diagnosed with advanced stage disease, squamous cell carcinoma of renal pelvis tended to early recurrence and metastasis and the prognosis was very poor.

OBJECTIVETo evaluate the association of sperm mobility parameters assessed by computer-assisted sperm analysis (CASA) with the rates of normal fertilization, oocyte cleavage and excellent embryos in in vitro fertilization (IVF) cycles.

METHODSA total of 288 infertile women undergoing IVF cycles patients were divided into two groups according to the normal fertilization rate (≥50% and <50%), cleavage rate (≥90% and <90%), or excellent embryo rates (≥50% and <50%). The means of the sperm motility parameters analyzed by CASA twice before oocyte retrieval were recorded and analyzed using t-test in relation to the rates of normal fertilization, cleavage and excellent embryos in IVF cycles.

RESULTSThe mean curvilinear velocity (VCL), average path velocity (VAP), and amplitude of lateral head displacement (ALH) were significantly higher in women with a normal fertilization rate of ≥50% than in those with a normal fertilization rate of <50% (P<0.05). Women with an oocyte cleavage rate of ≥90% had significantly higher VCL and VAP than those with a cleavage rate of <90% (P<0.05). The VCL, straight line velocity (VSL), VAP, linearity, straightness, wobble coefficient, ALH, or beat-cross frequency showed no significant differences between women with excellent embryo rates of ≥50% and <50% (P>0.05).

CONCLUSIONThe sperm motility parameters assessed using CASA are associated with normal fertilization and oocyte cleavage rates but not with excellent embryo rate in IVF cycles.

Adult ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; therapy ; Male ; Middle Aged ; Semen Analysis ; Sperm Motility ; Therapy, Computer-Assisted

OBJECTIVETo construct eukaryotic expression vectors for different domains (V and VC1) of the extracellular region of the receptor of advanced glycation end products (RAGE) and investigate the roles of these domains in prostate cancer.

METHODSThe coding sequence of V and VC1 domains was amplified from the plasmid pcDNA3-HA-RAGE by PCR and cloned into the pcDNA3-HA vector following routine procedures. After identification by PCR and sequencing, the vectors including V and VC1 domains were transfected into PC-3 cells. Western blotting and immunofluorescence were used to detect the expression and distribution of the expressed products in transfected PC-3 cells.

RESULTSThe expression vectors containing V and VC1 domains of RAGE were successfully constructed as confirmed by PCR and DNA sequence analysis. The V and VC1 domains of RAGE were highly expressed and showed a cytoplasmic distribution in transfected PC-3 cells.

CONCLUSIONThe constructed eukaryotic expression vectors for V and VC1 domains of RAGE can be efficiently expressed in prostate cancer cells.

Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; Humans ; Male ; Plasmids ; Prostatic Neoplasms ; genetics ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; Transfection