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Objective To evaluate the performance portable analyzer and reagent disc kit for liver function tests .Methods Ac‐cording to EP10‐A2 document of Clinic and Laboratory Standards Institute ,specimens with high ,medium and low concentrations were measured continuously in a specific order for 5 days .The bias and the total imprecision were calculated ,and the intercept ,slope rate ,non linearity ,carryover contamination and drift were analyzed by using multiple regression analysis .Results The bias and pre‐cision of specimens with high ,medium and low concentrations were all within acceptable ranges .No significant difference was showed in intercept ,slope rate ,non linearity ,carryover contamination and drift(P>0 .05) .Conclusion Portable analyzer and rea‐gent disc kit for liver function tests could be with fine precision ,fine linearity ,low carryover contamination and good stability ,and could meet clinical application requirement .
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Objective To investigate the molecular mechanisms of PLCεin regulating the invasion and migration of human bladder cancer cells in vitro.Methods After cells treated with recombinant adenovirus , the migratory/in-vasive abilities of T24 cells were explored by wound healing and Transwell chamber cell migration and invasion as -say;RT-PCR was used to detect the mRNA levels of PLCε;The protein levels of PLCε,PKCα,PKCβ, TBX3 and E-cadherin were determined by Western blot;QRT-PCR was used to detect the mRNA levels of TBX3 and E-cad-herin.Results It was confirmed by digesting and sequencing that the recombinant adenovirus had been constructed successfully .The expression of PLCε mRNA and PLCε protein were both decreased after the infection of Ad-shPLCε.Wound healing and Transwell chamber cell migration/invasion assay showed that Ad-shPLCε treatment could inhibit the migratory and invasive activity of bladder cancer cells(P<0.05).The results of Western blot indicated that the expression of PKCα/βin membrane decreased ( P<0.05 ) , and phosphorylation level of PKCαand PKCβwas reduced .QRT-PCR and Western blot analysis demonstrated that the expression level of TBX 3 de-creased , but the expression level of E-cadherin increased .Conclusions PLCε shRNA can inhibit migratory and invasive ability of bladder cancer cells through PKCα/β/TBX3/E-cadherin pathway .
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Objective To develop improved enzymatic creatinine(Cr)assay reagents (self-R&D),and to investigate their appli-cation on serum detection by comparing with imported commercial Cr reagents(enzymatic Cr reagents from Toyobo)Methods En-zymatic method was used to evaluate the effect of every component and different concentrations of reagents on Cr assay by detecting the alteration of absorbance of Cr before or after the reaction.Meanwhile,the blank absorbance and analysis sensitivity of self-R&D and imported reagents,the technical indicators of precision,linearity,as well as method comparison of self-R&D reagents,were de-tected on the same automatic biochemical analyzer.Results The blank absorbance of self-R&D reagents was 0.009,and the detec-tion sensitivity was 0.13,better than that of imported Cr reagents.The coefficient of variation (CV)of high and low values of ser-um of self-R&D reagents were 1.5%,and 1.1%,respectively.The linear range was 0-2 850 μmol/L and the method comparison result was Y =0.98X +1.15 (r =0.999).The expected bias was less than the allowable error region.Using relative deviation≥10% as an index to evaluate the existence of significant interference,it shows that 35 mmol/L of creatine,3.42 mmol/L of biliru-bin,0.03 g/L of vitamin C,5 g/L of hemoglobin and 1450 FTU chyle in both low and high concentration serum samples did not interfere with the test result.Conclusion The quality of self-R&D reagents was good,and there was a good relativity between self-R&D reagents and imported Cr reagents with excellent quality.This indicates the self-R&D reagents could satisfy the application requirements of the clinics.
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Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.
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Objective To evaluate urinary vascular endothelial growth factor (VEGF) in the diagnosis of bladder cancer. Methods VEGF in urine was measured by enzyme linked immunosorbent assay in 33 cases of transitional cell bladder carcinoma,10 benign urological conditions and in 10 normal individuals. Results The mean urinary VEGF level in 33 patients with bladder cancer was ( 309.8 ? 86.6 )ng/g Cr, significantly higher than in the 20 controls which was (107.6?35.4)ng/g Cr ( P
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Objective:To produce and identify antiCK20 monoclonal antibody.Methods:Lymphocytes from the spleen of mice being immunonized by CK20 antigen were fused with the myeloma cell line(SP2/0) using PEG4000.Hybridodma cells were established by selective growth of the fusion cells in the HT medium,and the presence of antiCK20 antibody was screened by inderect ELISA,and the clonality was achieved by limiting dilution.We have incubated cloning cells into mouse abdominal cavity to produce ascitic fluid contained monoclonal antibody.Chromatography with SPA-Sepharose CL-4B affinity column were emploied to isolate the monoclonal antibody from ascitic fluid.Finally,the antibody were tested the activities and sentivities,isoforms and titer through Western blot,two directions immuning diffusion of agar and ELISA.Results:Only one hybridoma cell line,secreating McAb against CK20,had been established.The modal number of chromosome is 101(99-103).The results of identifications showed that the antibodies kept high activitis and sensitivitis in detecting sample.The titer of ascitic fluid and the McAb purified are 1∶10~6 equally.The immunoglobulin of the McAb is classified as IgG1.Conclusion:AntiCK20 monoclonal antibody have been produced succesfully with high sensitive and active and was named L20031030.