Objective To observe the changes of neutrophil gelatinase associated lipocalin,NGAL after endoplasmic reticulum stress by thapsigargin in renal tubular epithelial cells.Methods The renal tubular epithelial cells were treated with thapsigargin to establish the model of endoplasmic reticulum stress (ER group).CHOP and GRP78 expression were detected by Western-blot to confirm endoplasmic reticulum stress in renal tubular epithelial cells by thapsigargin.NGAL expression were detected by Western-blot in ER group and control group(normal renal tubular epithelial cells).Results After treated with 2.5 μmol/L and 5 μmol/L thapsigargin for 4 hours and 8 hours respectively,endoplasmic reticulum stress in the renal tubular epithelial cells were significantly increased comparing with control group (0.585 ± 0.045 to 0.523 ± 0.030;0.785 ± 0.049 to 0.728 ± 0.064),which was approved by the increased expression of CHOP and GRP78 (P ＜ 0.05).The expression of NGAL was significantly increased in ER group (0.567 ± 0.024 to 0.826 ± 0.057,P ＜ 0.05) and it was corresponded with the changes of CHOP and GRP78,which increased significantly with the severity of endoplasmic reticulum stress.Conclusion Endoplasmic reticulum stress can be induced by thapsigargin in renal tubular epithelial cells.The expression of CHOP,GRP78 and NGAL were significantly increased correspondingly with the severity of endoplasmic reticulum stress.It is speculated that the expression of NGAL may be related with endoplasmic reticulum stress in tubular epithelial cells.

Alcoholism ; physiopathology ; prevention & control ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Male ; Maze Learning ; drug effects ; Memory Disorders ; prevention & control ; Prefrontal Cortex ; metabolism ; physiopathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Stilbenes ; pharmacology

Objective To establish a mouse model of prostate cancer expressing human PSCA for the development of new anti-tumor drugs or vaccines. Methods The total RNA of DU145 cells,a human prostate cancer cell line,was isolated by using TRIzol reagent according to the (RT-PCR),the first-strand cDNA was synthesized using the SuperScript First-Strand synthesis system. The human PSCA gene was amplified with the primers and cloned into the plasmid pcDNA3.1 to generate pcDNA-PSCA. DNA sequencing was used to confirm the constructs. The mouse prostate tumor cell line RM-1 cells,syngeneic to C57BL/6,were transfected with pcDNA-PSCA plasmids followed by selection using G418. RT-PCR analysis was performed to examine the validity of the constructs. Expression of PSCA on the cell surface was determined by staining with anti-PSCA antibody,and the anti-PSCA antibody was detected using an FITC-conjugated goat anti-rabbit IgG antibody,and analyzed by flow cytometry. 4-6-week-old male C57BL/6 mice purchased from the Laboratory Animals Center were inoculated with different amounts of RM-PSCA cells to search for suitable cell population which can form tumor in mouse,and the mice were monitored twice a week. The growth and the survival time of mice were measured,respectively. The tumor volume was measured by vernier caliper according to the formula:V=0.5a×b~2,where a and b are the long and short diameters of the tumor,respectively. Results The plasmid pcDNA-PSCA was successfully constructed and the PSCA was successfully expressed in RM-PSCA 7~# and RM-PSCA 28~# cells by RT-PCR and confirmed by flow cytometry. 1×10~5 RM-PSCA cells were sufficient to get tumor growth in 100% of inoculated mice. The tumor grew quickly and the volume of the tumor reached 12000 mm~3 within 34 days. All the mice died within 40 days and their mean survival time was 37 days. Conclusion A PSCA-expressing tumor model in mice has been successfully established. It can be used to evaluate the activities of drugs or vaccines.

AIM:To investigate the effects of down-regulated miR-9 expression on the proliferation , invasion and migration of nasopharyngeal carcinoma (NPC) cells.METHODS:Human NPC CNE1 and CNE2 cells were transfect-ed with the inhibitor of miR-9 by Lipofectamine to down-regulate the expression of miR-9, and the cells transfected with an inhibitor control were also set up .The cell proliferation and cell cycle were evaluated by CCK-8 assay and flow cytometry . The cell invasion and migration abilities were detected by Transwell invasion and wound -healing assays .Immunoblotting was applied to analyze the levels of the proteins .RESULTS:Compared with control group , inhibition of miR-9 expression in the NPC cells by transfection of the miR-9 inhibitor significantly decreased the proliferation ability (P<0.05).The per-centages of the cells in G 0/G1 phase [ CNE2: ( 57.96 ±1.39 )% vs ( 47.93 ±1.76 )%, P<0.05; CNE1: ( 51.24 ± 0.88)%vs (48.29 ±0.39)%, P<0.05] were significantly increased.The migration distances [CNE2: (186.50 ± 7.94)μm vs (247.56 ±15.56)μm, P<0.05;CNE1:(139.06 ±16.73)μm vs (230.66 ±14.27)μm, P<0.01] and the invasion ability of the CNE2 cells (43.00 ±3.17 vs 65.80 ±5.20, P<0.01) were also significantly inhibited .Moreo-ver, the tumor cells transfected with the inhibitors produced lower β-catenin.CONCLUSION:Inhibition of miR-9 expres-sion suppresses the proliferation , invasion and migration of nasopharyngeal carcinoma cells .

Objective: To compare the application of Sofren injection in respiratory department before and after the intervention to provide reference for clinical application and pharmacy management.Methods: A retrospective study was conducted on the medical records of inpatients treated with Sofren injection, and then intervene the found problems.Comparative study of Sofren injection in respiratory department before and after the intervention was performed.Results: Among the 584 cases in the retrospective study, those with off-indications were 438 ones (75%), those with inappropriate solvents were 555 ones (95.03%), and those with improper course were 196 ones (33.56%).After the intervention in the clinical practice, the improvement rate of improper solvents was 98.70%, that of off-indications was 29.85%, and that of unsuitable course was 77.71%.Conclusion: Through comparative study on the main problems before the intervention and the effective performance of specific solutions, the application of Sofren injection in respiratory department is greatly improved, which provides reference for the special management of other drugs.

OBJECTIVETo evaluate effect of root canal curvature and location of the fragment on the removal of broken file from root canal.

METHODSSixty extracted mandibular premolars were divided equally into six groups according to location of fragment (2 mm or 8 mm below root canal orifice) and root canal curvature (20 degrees, 30 degrees or 40 degrees). Broken files were removed using ultrasonic tips combined with dental operating microscope. Number of successfully removed case and operating time were recorded. Pre- and post-operative digital radiographs were input into image analyzing software to calculate diameter variance of root canal at the level of tip of broken file.

RESULTSAll the 6 groups of broken files were successfully removed with out perforation. Among same fragment location groups, diameter variance was increased in greater curvature groups and the difference was significant (P < 0.05), while operation time was not significantly different (P > 0.05). Among same root canal curvature groups, the deeper the fragment, the longer operation time, difference being significant (P < 0.05), while difference of diameter variance was not significant (P > 0.05).

CONCLUSIONThe more curvature of root canal, the more dentine removal amount at the level of broken file tip. Location of fragment has no effect on dentine removal amount. Safe straight-line access and adequate thickness of dentine are essential to removal of broken file from root canal.

Bicuspid ; Dental Pulp Cavity ; Dentin ; Humans ; Root Canal Preparation ; Root Canal Therapy

Objective To express human-mouse chimeric antibody against anthrax protective anti-gen and to analyze its biological activities. Methods A new mammalian bipromoter expression vector was constructed with dihydrofolate reduetase(DHFR) gene as the selection and complication marker. First, the light and heavy chain variable region gene of the monoclonal antibody 5E1 were cloned by RT-PCR, at the same time the human IgG1 heavy chain constant region gene and kappa type constant region gene were cloned. Next, the human-mouse chimeric antibody genes were synthesized by fusion PCR. Then, the hu-man-mouse chimeric antibody gene were inserted into MCS of pSecTag and B1 to construct pSecTag-5E1L and B1-5E1H, respectively. Finally, heavy chain expression cassette excised from the B1-5E1H with Bgl Ⅱ/BamH Ⅰ was further cloned into the Bgl Ⅱ site of the pSecTag-5E1L to construct pSecTag-5E1. Plasmid pSecTag-5E1 was transfected into CHO(dhfr) engineering cells and high production cell clones that were screened by enhancing MTX concentration. After collecting medium and purifying chimeric antibody with af-finity chromatogram, purified chimeric antibody was analyzed by SDS-PAGE, Western blot. Results A sta-ble and high production cell line was acquired at MTX concentration 5×10~(-8) mol/L. Conclusion The hu-man-mouse chimeric antibodies were successfully expressed in CHO cells.

Objective To analyze and compare the imaging features of chromophobe cell renal carcinoma(CCRC)with pathologic findings in order to improve the diagnostic accuracy.Methods The data of CT and MRI of 12 patients with CCRC were reviewed retrospectively.Ten patients underwent CT examination,including precontrast scan,the contrast eortieomedullary phase scan and the parenchymal phase scan(one patient without corticomedullary phase scan).Two patients underwent MR examination including precontrast T_1WI,T_2WI and enhanced T_1WI of the corticomedullary phase and the parenchymal phase.Results Four lesions located in left kidney and eight in right kidney.Maximum diameter of lesions ranged from 24 mm to 125 mm,average 56.7 ram.Homogenous density was observed in six lesions of ten on unenhanced CT scan and five lesions had homogenous enhancement on enhanced CT scan,which was due to the less incidence of necrosis,liquefaction and hemorrhage on pathologic findings.Nine Lesions showed hyperdense compared with renal medulla but the density was lower than renal cortex on the corticomedullary phase.The enhanced degree was positively correlated with microvessel density(MVD).All ten lesions became hypodense compared with renal medulla on the parenchymal phase scan.Central stellate scar was found in two big lesions and psudocapsula were observed in four lesions confirmed by pathology.Two patients underwent MRI examination.Compared with medulla,the two lesions showed hyperintense on unenhanced T_1WI and obviously hypointense on unenhaneed T_2WI.The enhancement pattern of them was similar to CT. Conclusion The imaging features of CCRC,such as homogeneity,special enhancement pattern and distinct hypointensity on T_2WI,help to differentiate CCRC from other renal tumors.

OBJECTIVETo evaluate the nerve remodeling induced by 48 hours right atrial pacing in a canine model.

METHODSRapid right atrial pacing (600 beats/min) was performed in 6 mongrel dogs of either sex for 48 hours to induce sustained atrial fibrillation (AF). Six dogs without pacing served as controls. Cardiac nerves were immunocytochemically stained using anti-growth-associated protein 43 (GAP43) and anti-choline acetyltransferase (CHAT) antibodies to compare nerve sprouting and pneumogastric nerve remodeling between the 2 groups.

RESULTSIn dogs with AF, the GAP43-positive and CHAT-positive nerve densities in the left atrium, left auricular appendage, right atrium and right auricular appendage were significantly higher than in control animals (all P < 0.05). Moreover, nerve density was significantly higher in the right atrium than in the left atrium in dogs with AF. Microscopic examinations revealed an inhomogeneous distribution of cardiac nerves.

CONCLUSIONSignificant nerve sprouting and pneumogastric nerve remodeling were evidenced in the right and left atrium in a canine model of sustained AF induced by 48 hours right atrial pacing.

Animals ; Atrial Fibrillation ; physiopathology ; Cardiac Pacing, Artificial ; Disease Models, Animal ; Dogs ; Female ; Heart Atria ; innervation ; physiopathology ; Male ; Vagus Nerve ; physiopathology