ObjectiveTo examine the effects of temsirolimus, an inhibitor of mammalian target of rapamycin, on bladder cancer cell lines T24 and BIU-87 in vitro and in vivo for purpose of evaluating the probability of mTOR targeted therapy for bladder cancer.MethodsAfter being treated by a different concentration of temsirolimus, T24 and BIU-87 cells were tested by MTT assay for cell proliferation activity.Cell cycle and apoptosis analysis were performed with flow cytometer. Wound scratch assay was used for cell migration activity and transwell motility assay. Western blot analysis was used to test the mTOR phosphorylation. Subcutaneous inoculation of 6-week-old nude mice was performed using 1 × 106 T24 cells in 50％ matrigel for both control (n = 10) and temsirolimus (n = 10) groups. The volume of tumors was examined and then the expression of Ki-67 was detected by immunohistochemistry.ResultsTemsirolimus significantly inhibited proliferation of T24 and BIU-87 cells in a dose- and time-dependent manner. After administration of temsirolimus on T24 and BIU-87 cell lines for 24 h, the rate of wound healing in 0 nmol/L groups were (88.9 ± 14. 1 ) ％ and ( 83.6 ± 16.3)％ , which were higher than in the 5 nmol/L groups, which were (42.7 ± 11.6) ％ and ( 36.9 ± 9.7 ) ％ ( P ＜ 0.05 ). In the transwell motility assay, the number of cells in the 0 nmol/L group was 26.5 ± 5.8 and 28.2 ± 4.6, which was higher than in the 5 nmol/L group ( 19.0 ±3. 8 and 21.3 ± 5.1, respectively) (P ＜ 0. 05). When temsirolimus was administered on T24 and BIU-87 cell lines for 48 h the percentages of cells delayed in phase G0/G1 in 5 nmol/L group were ( 77.46 ±6.11)％ and (73. 39 ± 4. 94)％ respectively, and higher than in the 0 nmol/L group, which were (65.99 ±5.01 )％ 、(60.15 ±3.98)％ (P ＜0.05). There was no statistically significant difference in the apoptosis rate between the two groups (P ＞ 0.05 ). In Western blot analysis, the ratios of p-mTOR/β-actin were 0.92 ±0.09 and 1.01 ± 0.08 in 0 nmol/L group, and higher than in the 5 nmol/L group (0.47 ±0.05、0.04 ±0. 01 ) (P ＜ 0.05 ). After administration of temsirolimus for 21 days, the tumor volume in nude mice in the control group were 351.1 ± 139.9 mm3 , which was larger than 351.1 ± 139.9 mm3 in the temsirolimus group ( P ＜ 0.05 ). The positive rate of Ki-67 expression was ( 67.3 ± 8.4 ) ％ in the control group, which was higher than in the temsirolimus group ( 35.5 ± 6.7 ) ％ ( P ＜ 0.05 ).ConclusionsThis study provides in vitro and in vivo evidence that temsirolimus may inhibit the viability of bladder cancer cells and temsirolimus could be exploited as a potential therapeutic strategy in bladder cancer.

To study the effects of Qinggan Huoxue Recipe (QGHXR), the compound traditional Chinese herbal medicine, and its separated recipes on the expression of tumor necrosis factor-alpha (TNF-alpha) mRNA and serum TNF-alpha content in rats with alcoholic liver injury (ALI).

Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.

Objective To determine the main CT features and the key points of differential diagnosis of multilocular cystic renal cell carcinoma (MCRCC) classified according to 2004 WHO pathological diagnostic criteria. Methods According to the criteria, 40 patients were divided into two groups: MCRCC group and other subtypes of cystic renal cell carcinoma (CRCC). The CT findings were evaluated and compared between two groups for cystic content, wall, septum, nodularity, calcification and enhancement. ROC curve was used to determine the cut-off value of the possible CT feature which could distinguish MCRCC from other subtypes of CRCC. Results Seventeen cases of MCRCC group and 23 cases of CRCC group were included in this study according to the diagnostic criteria. MCRCC appeared as a well defined multilocular cystic mass with thin wall and sepia and no expansile solid nodules. Thickness of cystic wall and/or septum is was main CT findings to distinguish MCRCC from other subtypes of CRCC (P ＜ 0.01 ). The cut-off value of the thickness was 6 mm and its sensibility, specificity was 89% ,75% respectively. Conclusion Cystic wall and/or septum with a thickness of less than 6 mm are the main CT findings to dis tinguish MCRCC from other subtypes of CRCC.

Objective To report 1 case with renal immunoglobulin light and heavy chain deposited disease (IgG-κ light chain and γ1 heavy chain).Methods The clinical manifestations,serum immunofixation electrophoresis,light and heavy chain abnormalities of blood and urine,bone marrow biopsy and renal biopsy data laboratory data were recorded and analyzed.Results A 63-year-old woman presented with massive proteinuria,microscopic hematuria,hypertension,anemia and serum IgG-κ light chain.Bone marrow aspirate revealed 4％ plasma cells.Renal biopsy revealed nodular glomerulopathy with congo red staining-negative.Immunofluorescence showed that κ light chain and IgG1 (γ1 heavy chain) were deposited along the glomerular basement membranes (GBM) and tubular basement membranes (TBM).Electron-microscopy revealed electrondense deposits that delineate the outer aspect of TBM and endothelial aspect of GBM.Conclusion The diagnosis of renal immunoglobulin light and heavy chain deposited disease (IgG-κ light chain and γ1 heavy chain) should be addressed combine with clinical and is pathology,especially with immune pathological examination.

Adolescent ; Adult ; Child ; Female ; Heart Failure ; blood ; Humans ; Male ; Middle Aged ; Myocardium ; metabolism ; Serum Response Factor ; metabolism

Animals ; Female ; Injections, Intraventricular ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; Mice, Inbred Strains ; Toluene ; administration & dosage ; toxicity

Objective To assess left atrial reservoir function based on left atrial filling volume,the left atrial expansion index and left atrial ejection fraction (LAEF) were measured in patients with left ventricular non-compaction cardiomyopathy (LVNC),using real-time three-dimensional echocardiography (RT3DE),to determine the value for LVNC diagnosis.Methods The study included 20 patients diagnosed with LVNC,including 3 patients,aged 4-16 years;8 patients,aged 17-35 years;9 patients,aged 36-55 years;and 20 healthy age-matched control subjects.All patients underwent two-dimensional echocardiography and RT3DE.Results Two-dimensional echocardiography showed no differences between groups (P ＞ 0.05).RT3DE analysis showed that the average left atrial filling volume/BSA,left atrial expansion index,and LAEF were significantly higher in the LVNC group.Conclusion LVNC is associated with increased left atrial reservoir function.RT3DE shows promise for the diagnosis of LVNC.

Objective To investigate the correlations between plasma levels of carnitine(CT),free fat acid(FFA) and insulin resisitance in children with simple obesity.Methods Fifty-six children diagnosed with simple obesity were enrolled as study group(obesity group),36 healthy children as control group.The concentration of plasma level of insulin was measured by radioimmunoassay(RIA),CT was measured by high performance liquid chromatography(HPLC),FFA and triglyceride(TG) were measured by enzymatic-colorimetric assay.Body mass index(BMI) and waist to hip ratio(WHR) were caculated.Insulin sensitivity index (InSI) and insulin resistance index (InRI) were cacula-ted by homeostasis model assessment of insulin resistance (HOMA-IR).SPSS 13.0 software was used to analyze the data.Results The concentration of CT in plasma was (43.67?12.75) ?mol/L in obesity group,(58.31?21.25) ?mol/L in control group,respectively.There was a significant difference between 2 groups (t=2.566 P

AIM:To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tis-sues and the corresponding paratumorous tissues collected from 63 patients.The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured .The cell viability was measured by CCK-8 assay.Flow cytometry was used to monitor the changes of cell cycle and apoptosis .The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS:The expressed level of miRNA-363 was lower , and the expression level of SOX 4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues .A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed .The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated , with significant difference as compared with the cells transfected with miRNA-NC and control cells .The viability of MG-63 cells was inhibited, the cell cycle was arrested in G0/G1 phase, and the cell apoptosis was increased by transfec-tion with miRNA-363 mimics.The relative protein expression levels of SOX 4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group , but the relative expression levels of miRNA-363 had no significant difference .Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CON-CLUSION:The expression level of miRNA-363 is low in human osteosarcoma tissue .miRNA-363 may inhibits the viabili-ty of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.