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.

Adolescent ; Child ; Child, Preschool ; Humans ; Infant ; Infant, Newborn ; Rickets ; prevention & control ; Vitamin D ; therapeutic use

Objective To evaluate the value of color Doppler flow imaging (CDFI) in the diagnosis of elderly femoral arterial pseudo aneurysm and treating with ultrasound-guided compression after percutaneous transluminal coronary angioplasty (PTCA). Methods Twenty-six elderly patients who were found a mass and/or vascular murmur in the puncture region were examined by CDFI, and the results of CDFI were proven by clinic and MRI. All pseudo aneurysms were repaired by CDFI guidance to compress simply and/or injecting directly batroxobin into the pseudo aneurysm lumen combined with compression.Results Pseudo aneurysms were diagnosed definitely by CDFI in 26 cases,and the accuracy rate was 100%. Twenty-two cases were repaired with CDFI guidance to compress simply and percutaneously injection of batroxobin into the pseudo aneurysm lumen combined with compression occurred in 3 cases. One case was cured by surgery after failing to be repaired with CDFI guidance. Conclusions CDFI has an important diagnosis value of pseudo aneurysm after PTCA. The method of CDFI guidance compression and batroxobin injection for repairing pseudo aneurysm is simple, safe and effective.

Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P ＜ 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P ＞0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P ＜0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P ＜0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P＜0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.

Humans ; Hypertrophy ; diagnostic imaging ; Infant, Newborn ; Liver ; diagnostic imaging ; Male ; Tomography, X-Ray Computed

BACKGROUND:Chitin has been found to be a good biomaterial, but research on chitin carrying corneal epithelial cel s for rabbit corneal epithelial injury is little reported. OBJECTIVE:To investigate the repair outcomes of chitin hybrid membrane carrying corneal epithelial cel s in the rabbit corneal epithelial injury.METHODS:Eighteen New Zealand white rabbits were enrol ed and made into left corneal epithelial injury models, and then randomized into two groups and treated with chitin hybrid membrane carrying corneal epithelial cel s (experimental group) and chitin hybrid membrane (control group), respectively. The damage area, histological changes and ultrastructure of the cornea were observed at 1, 3, and 7 days after implantation. RESULTS AND CONCLUSION:Damage area of the cornea in the experimental group was significantly less than that in the control group at 1 and 3 days after implantation (P<0.05), and the cornea in both two groups healed wel at 7 days after implantation. At 7 days after implantation, in both two groups, the corneal epithelium with six layers adhered to the corneal stroma closely, which was repaired completely and regularly. Comparatively speaking, the cornea in the experimental group possessed smooth outer layer. Besides, in the experimental group, the hexagonal corneal epithelial cel s arranged closely with flat surface;while the hexagonal corneal epithelial cel s in the control group showed no smooth surface and gaps between cel s. These results indicate that chitin hybrid membrane carrying corneal epithelial cel s promotes the repair of rabbit corneal epithelial injury.

BACKGROUND:Chitosan nanoparticles-encapsuled sodium hyaluronate is an effective drug for the burned cornea. OBJECTIVE:To verify the effect of sodium hyaluronate/chitosan nanoparticles on the neovascularization in burned cornea. METHODS:Thirty Sprague-Dawley rats were randomly divided into three groups, and the model of burned cornea caused by base was established in the rats of model and experimental groups, fol owed by respectively treated with 10μL sodium hyaluronate/chitosan nanoparticle suspension and normal saline, once daily, for consecutive 4 weeks. Rats only given normal saline were used as controls. Four weeks later, the dynamic growth of newly formed blood vessels in the cornea was observed using silt lamp. The levels of tumor necrosis factor-αand interleukin-6 were detected by ELISA, histological changes of the cornea were observed by hematoxylin-eosin staining, and the mRNA expression levels of vascular endothelial growth factor and cyclooxygenase 2 were detected by real-time PCR. RESULTS AND CONCLUSION:Compared with the control group, the area of the newly formed blood vessel and the levels of tumor necrosis factor-α, vascular endothelial growth factor and cyclooxygenase 2 were significantly increased in the model group (P<0.05, P<0.01). In the experimental group, al above indicators were significantly lower than those in the model group (P<0.05). There were a large number of inflammatory cel s and neovascularization in the model group, but only few inflammatory cel s in the experimental group. These results show that sodium hyaluronate/chitosan nanoparticles can inhibit the neovascularization in the burned cornea.

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.

Objective To investigate the application of quantitative parameters associated with iodine concentration derived from iodine overlay image by dual-source dual-energy computed tomographic (CT) in differentiating benign and malignant thyroid nodules. Methods Seventy-eight patients (total 112 nodules, including 64 benign and 48 malignant nodules) with thyroid nodules who underwent plain scan (PS), arterial phase (AP) and venous phase (VP) enhanced scan by DSCT (80 kVp/ Sn140 kVp) were analyzed retrospectively. Iodine overlay images were obtained by the dual energy post-processing software. The mean iodine concentrations in the normal thyroid parenchyma (ICThy), the lesion (ICLes), and the carotid artery(ICCar) of PS, AP and VP were measured from iodine overlay images. The iodine concentration difference (ICD), the iodine concentration difference-to-normal parenchyma ratio (ICDNR) and the normalized iodine concentration ratio (NIC) were calculated. The two-sample t test was performed to compare quantitative parameters between the benign and malignant nodules. ROC curve with quantitative parameters of three phases was used to analyze the diagnostic efficiency of ICD, ICDNR, NIC and ICles. Results During
PS, mean ICDNR, ICD and ICLes of malignant nodules were respectively 1.04 ± 0.95, (2.20 ± 1.82) mg/ml, (-0.04 ± 1.65) mg/ml, ICDNR, ICD and ICLes of benign nodules were respectively 0.04 ± 0.41, (0.35 ± 0.97) mg/ml, (2.19 ± 0.55) mg/ml. ICDNR and ICD of malignant nodules were higher than benign nodules (t'=6.63, 6.39, P<0.05), while ICles of malignant nodules were lower than benign nodules (t=10.13, P<0.05). During AP , mean ICDNR, ICD, ICLes of malignant nodules were 0.39 ± 0.29, (2.23 ± 1.77) mg/ml, (3.81 ± 1.50) mg/ml, and benign nodules were 0.49 ± 0.22, (2.97 ± 1.91) mg/ml, (3.17 ± 1.64) mg/ml, respectively. ICDNR, ICD of malignant nodules were lower than benign nodules (t'=2.08, t=2.12;P<0.05),while ICles of malignant nodules were higher than benign nodules (t=2.12, P< 0.05). During VP, mean NIC of the malignant and benign nodules were 0.45 ± 0.21, 0.58 ± 0.37, respectively. NIC of malignant nodules were lower than benign nodules (t'=2.35, P< 0.05). AUC of ICDNR during PS was 0.892, the sensitivity was 83.3% and the specificity were 90.5%. Conclusion Quantitative parameters associated with iodine concentration by dual-source dual-energy CT may increase the efficiency and accuracy in differentiating benign and malignant thyroid nodules.

Objective To analyze the influencing factors of deep vein thrombosis (DVT ) in the patients with abdominal trau‐ma .Methods A total of 200 cases of patients with abdominal trauma in our hospital from June 2014 to June 2015 were selected and performed the color Doppler ultrasonography for determining whether DVT occurring .The clinical data were retrospectively ana‐lyzed .The influencing factors of DVT occurrence in the patients with abdominal trauma were analyzed .Results Among 200 cases of abdominal trauma ,56 cases appeared the symptoms and signs of muscular pressing pain ,swelling pain ,positive Homans sign ,su‐perficial varicose and skin temperature decrease within 7 d after abdominal trauma ,60 cases were diagnosed as DVT by color Doppler ultrasonography .The multivariate Logistic regression analysis results showed that advanced age ,high cholesterol level ,high D‐dimer level ,high blood urea nitrogen level ,complicating hypertension ,complicating diabetes ,complicating hyperlipidemia ,surgical history at preoperative 1 month ,lower abdominal trauma ,high score of trauma ,bedridden time more than 3 d and infection were the independent risk factors of DVT occurrence in abdominal trauma patients (P<0 .05) .Conclusion Clinic should perform the color Doppler ultrasound screening in the abdominal trauma patients with risk factors of DVT ,which is conducive to early discovery and early treatment of DVT .