Objectives:To investigate the effects of small interference RNA(siRNA) silencing Annexin Ⅱ on invasion and metastasis of human breast cancer cell MDA-MB-231.Methods:The chemically-synthetic siRNA duplexes targeting Annexin Ⅱ(Annexin Ⅱ-siRNA) was transiently transfected into MDA-MB-231 cells which have high metastatic potential by lipofectamin 2000.The transfection efficiency was observed by fluorescence microscopy.RT-PCR and western blot were used to semi-quantify the Annexin Ⅱ mRNA and protein levels.The malignant characters of transfected MDA-MB-231 cells including celluar proliferation rate,the activities of invasion and metastasis were analyzed by use of 3-(4,5-dimethylthiazol-2-yl)-5-diphenyl tetrazolium bromide(MTT) and millicell chamber assay,respectivilly.Results:Annexin Ⅱ-siRNA effectively inhibited Annexin Ⅱ mRNA and protein levels(P

Objective To establish a SYBR Green based real-time fluorescence quantitative PCR method for detecting human Annexin Ⅱ mRNA expression,and to detect the level of Annexin Ⅱ mRNA in human breast cancer cells MCF-7 and MDA-MB-231.Methods The specific primers were designed according to the conserved sequence of human Annexin Ⅱ gene.Total RNAs were extracted from human breast cancer cells(MCF-7,MDA-MB-231),then RNAs were transcribed reversely into cDNAs.The plasmid standards were constructed.The relative expression levels of human Annexin Ⅱ mRNA in human breast cancer cells were detected by this method.Results The square(r2 )of correlation coefficient of the standard curve in this method was 0.997,the melting curve analysis showed the single peak.The the intra-batch and inter-batch variable coefficients in the pGM-T Annexin Ⅱplasmid standard substance were 6.2%,7.8% and 9.1%,12.3% respectively.The further study indicated that AnnexinⅡ mRNA in MDA-MB-231 was higher than that in MCF-7(P<0.01).Conclusion The established SYBR Green real time fluorescence quan-titative PCR method for detecting human AnnexinⅡ is of good specificity and repeatability and can be used for quantitatively detec-ting AnnexinⅡ mRNA in breast cancer cells.

Objective To explore a stable and feasible method for the primary culture of breast cancer-associated fibroblast (CAF), and to offer the theoretical basis for the further studies about the role of CAF in the processing of breast cancer by analyzing the biological property of CAF. Methods CAF was primary cultured with tissue block enzymolytic method. Flow cytometry (FCM) was used to detect the cell cycle. The expression of Vimentin, α-SMA and Ki-67 were detected by cell immunochemistry method. Transwell assay was used to detect the invasion and migration capacity of CAF. Results The CAF was successfullyculturedbyconditionoptimization.ImmunohistochemicalresultsshowedthatVimentin, α-SMA and Ki-67 were highly expressed in CAF.Comparedwithfibroblastsfrombreastcancersidestissues,the proliferating ability and the invasion and migration capacities of CAF were enhanced (number of invasive cells: 79.7 ± 3.5 vs. 66.3 ± 1.5, number of migrated cells: 242.3 ± 3.1 vs. 218.3 ± 2.1, both P<0.05). Conclusion CAF cultured from breast cancer has better proliferating ability and strong invasion and migration capacities.

Objective To evaluate the performance of UW2000 automatic urine system .Methods Using UW2000 automatic u‐rine system ,to evaluate intra ,inter batch precision ,linear ,carryover ,and the accuracy of the urine visible components(RBC ,WBC);at the same time intra ,inter batch precision and accuracy of of urine dry chemistry component .Results The within‐run coefficients of variation(CV% ) of RBC in high ,middle and low‐value samples were 15 .6% ,6 .8% ,20 .3% ,WBC were 16 .4% ,10 .5% ,24 .6% , respectively .Inter‐run precision of variation(CV% ) of RBC in middle and low‐value were 12 .8% ,13 .2% ,WBC were 15 .3% , 22 .8% ,respectively .The correlation coefficient of RBC was 0 .991 and the linear range was 88-19 513/μL .The correlation coeffi‐cient of WBC was 0 .997 and the linear range was 7-8 254/μL .And carryover rate was 0 .00% for RBC and 0 .06% for WBC ;The coefficient of UW2000 and microscopy on RBC and WBC were 0 .992 ,0 .995 .Within‐run precision ,inter‐run precision and accuracy for urine dry chemical composition of high ,low controls were all within the requirements of standards .Conclusion UW2000 auto‐matic urine analyzer workstation was not only save manpower ,but also with high detection speed and save cost .Importantly ,the re‐sults are reliability .In addition ,it could meet the working requirement in the general hospital ,and has great application value .

Objective To observe the turnaround time(TAT) of biochemistry laboratory in a certain hospital of Chongqing city,and to improve the quality of the laboratory by shortening the TAT.Methods TAT was analyzed by analyzing the daily workload,average TAT and failure rate of outpatient clinics,outpatient emergency,inpatient clinics,and inpatient emergency subjects from 2013 to 2015.The reasons for TAT prolongation were analyzed.Results The biochemical test samples were 77 060,97 129 and 105 304 from 2013 to 2015,and the annual growth rate was 26.0％ and 8.4％ respectively.TAT of the routine outpatient department samples were (78.55nu48.47)、(69.18± 37.20)、(62.82 ±21.62)min,which decreased year by year,and the difference were statistically significant(P＜0.05),and the TAT of the outpatient emergency were (64.13 ± 31.16),(59.22 ± 23.51),(66.01±37.73)min.TAT of inpatient clinics were (92.34± 53.41),(95.03±55.73) and (122.92±78.94)min from 2013 to 2015,which increased year by year,and the difference were statistically significant(P＜0.05),and the TAT of the inpatient emergency were(65.29±36.06),(62.41±30.18),(61.48±30.12)min,which decreased year by year,and the difference were statistically significant (P＜0.05).The substandard rate of samples aforementioned were 0.04％,2.99％,0.63％ and 3.69％,respectively.Conclusion TAT increases with the samples increase,it is necessary that making sure staffs more responsible in daily work,optimizing the procedure of daily biochemistry tests,improving ability of serving for clinic and patients.

Objective To analyze the correlation between thrombelastography (TEG) and conventional coagulation indexes in breast cancer patients, and to compare the differences between the two methods in the detection of coagulation function in breast cancer patients.Methods Retrospectively analyzed the clinical data of180patients with breast cancer who were performed TEG, coagulation and blood test in the same day in our hospital from November 2016to May 2017.Linear correlation and regression analysis were performed among the parameters.The differences of positive rates of TEG parameters and conventional coagulation indexes were compared.Results The R value of TEG parameters of breast cancer patients was positively correlated with K and APTT, and negatively correlated withα-Angle, MA, CI, DD, FDPs.K was positively correlated with APTT and TT, and negatively correlated withα-Angle, MA, FIB, DD, FDPs and PLT.α-Angle was positively correlated with MA, CI, FIB, DD, FDPs and PLT.MA was positively correlated with CI, FIB, DD, FDPs and PLT.CI was positively correlated with FIB, DD, FDPs and PLT.α-Angle, MA and CI were all negatively correlated with APTT and TT, the difference were statistically significant (P＜0.05).A linear regression equation of TEG parameters and coagulation indexes was obtained.There was no significant difference between TEG detection positive rate and conventional coagulation test (P＞0.05).Conclusion There are significant correlation between the TEG parameters and routine coagulation or platelet, and the results are consistent.

Objective To analyze the correlation between thrombelastography(TEG)and coagulation, platelet count(PLT)in patients with malignant tumor.Methods Retrospectively analyzed the clinical data of 241 cases with tumor who were performed TEG,coagulation and blood test in the same day in Chongqing Cancer Institute from November 2016 to March 2017.Linear correlation and regression were carried out to an-alyze relationship among the parameters.The number of patients with positive blood clotting,PLT and TEG parameters were counted,and the χ2test was used to compare the difference between them.Using Mann-Whit-ney U test to compare the differences between multiple parameters of liver cancer,breast cancer,and pancreatic cancer.Results The R value of TEG parameters in patients with malignant tumor was positively correlated with APTT,negatively correlated with TT,DD and FDPs.K was positively correlated with APTT and TT, and negatively correlated with FIB and PLT.The relationship between α and FIB,PLT were positive,between APTT and TT were negative.MA was positively correlated with FIB and PLT,negatively correlated with TT, CI was positively correlated with FIB and PLT,and negatively correlated with APTT(P<0.05).The correla-tion between FIB,PLT and MA was the highest.And the linear regression equation of TEG parameters and coagulation indexes was obtained.The positive rate of TEG was lower than that of coagulation(P<0.05). Same certain differences of TEG and coagulation parameters were existed in liver cancer,breast cancer and pancreatic cancer patients.Conclusion TEG is significantly associated with PLT and conventional coagulation test,and the results of TEG and conventional coagulation test are consistent to a certain degree,but the overall agreement is generally not interchangeable.TEG might be play a complementary role with coagulation tests and platelet counts.And the TEG of different cancer types is not exactly the same as the coagulation parameters.