Objective To investigate the situation of cognitive demands for disease of discharged patients with T tube,in order to adopt a more rational and effective interventions to meet the needs of patients.Methods 60 patients were investigated with the questionnaire survey designed to gauge patients'cognitive demands for disease and health knowledge.Results The patients had higher cognitive demands for T tube retention time,how to make self-observation and influence of T tube retention on normal work and life,among these patients,they laid different particular emphasis to some items between different sexes.Conclusions Nurses can provide patients with comprehensive,prioritized and individualized health guidance according to cognitive demands of patients,so that the health education quality and work efficiency can be improved.

Objective To investigate the testing capabilities of clinical transfusion laboratories in medical institutions in Beijing for the regulatory authorities to formulate administrative policies in this regard.Methods Experts assigned by Beijing Clinical Transfusion Quality Control Center made on-site inspections at the transfusion laboratories in medical institutions using quality control products.They recorded the complete testing process of the operators as well as the instruments,detection reagents in use and the testing results,with statistics and analysis made to the data so collected.Results The pass rate of these on-site inspections was lower than that of the external quality assessment.Some laboratories failed to complete the testing of the quality control products in time and the actual operations in some laboratories were inconsistent to the guidelines.55.9％ of level Ⅰ hospitals and 25.6％ of level Ⅱ hospitals were found with insufficient and inadequate instruments and process layout to meet the needs of clinical blood transfusion.Some of the technicians were found without sufficient trainings in their professional knowledge and basic skills,resulting in their poor competence against emergency cases and weakness in independent problem solving.In addition,the records of detection process and results were found to be substandard.Conclusions Transfusion laboratories in Beijing need to improve their testing capabilities in general.

Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.

ObjectiveTo investigate the event-based prospective memory (EBPM) and time-based prospective memory (TBPM) in patients with Alzheimer' s disease (AD). MethodsTwenty patients with AD, 20 adults with amnesia mild cognitive impairment (aMCI) and 30 healthy adults with matched age and education level were assessed with a battery of neuropsychological tests including EBPM and TBPM tasks.ResultsCompared with healthy elders and patients with aMCI on performance of PM (2. 23 + 0. 77,4.83 ±1.09;1.00±1.03,3. 10 ± 1.52) and episodic memory(0. 70 ±0. 12,0.66 +0. 16;0.45 ±0.07,0.54±0. 10), AD patients were all impaired in PM and episodic memory(0.20 +0.41,2.05 ± 1.43;0.33±0. 12,0.32±0. 10), and were impaired in EBPM more significantly (t=-2.792, P＜0.01;t =-10. 761 ,P ＜0. 01 ). ConclusionsThese results suggest that AD patients show deficits of PM, but their EBPM is impaired more significantly. EBPM impairment may be an early diagnostic of AD.

Objective To study the effects VEGF small interfering RNA (siRNA) on chemosinsitivity of human BxPC3 cell and its mechanism. Methods BxPC3 cells were divided into single BxPC3 cell group,lipofection group, scrambled siRNA transfection (200 nmol/L) group, VEGF siRNA transfection group.VEGF siRNA (5, 10, 20, 100,200 nmol/L) was used to transfect BxPC3 cells. Expressions of VEGF mRNA and protein were determined by real-time PGR and ELISA assay, respectively. MTT was performed to examine the inhibitory effects of gemcitabine on BxPC3 cells of each group. The phosphorylated-Akt protein was evaluated by Western blotting. Results After VEGF siRNA transfection, the expression of VEGF mRNA and protein in BxPC3 cells was down-regulated in a dose-and time-dependent manner, but there was no effect on BxPC3 cells proliferation. After 0. 2 μmol/L of gemcitabine treatment for 48 h, the inhibitory rates were ( 16.9 ±0.3)%, (17.3 ±0.3)%, (28.8 ±0.4)%, (52.2 ±0.3)%, (75.4 ±0.4)% in BxPC3 cell group,lipofection group, 5,10,20 nmol/L VEGF siRNA transfection group, and the inhibitory effects were correlated with siRNA concentration ( r = 0. 928 ). The phosphorylated-Akt protein was reduced significantly in siRNA transfected cells. Conclusions VEGF gene plays an important role in BxPC3 cells resistence to chemotherapy through inhibiting Akt phosphorylation.

OBJECTIVE:To study the effects of paclitaxel combined with cisplatin on the proliferation,migration and invasion of thyroid cancer cells SW579 and its mechanism. METHODS:Cells were divided into blank control group,paclitaxel group (3μmol/L),cisplatin group(30 μmol/L),drug combination group(paclitaxel 3 μmol/L+cisplatin 30 μmol/L). 48 h after culture,the relative cell activity was measured by MTT assay. Cell cycle was detected by flow cytometry. Migration and invasion of cell was tested by Transwell assay. The expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN),protein kinase B(AKT),Cyclin D1,p27,matrix metalloproteinase(MMP)-2 and MMP-9 were detected by Western blot. RESULTS:Compared with blank control group,relative cell activity of all treatment groups were decreased;paclitaxel or plus cisplatin also made cell cy-cle arrest in G1 phase,and migration and invasion ability of cell were decreased;the expression of PTEN and p27 remarkably in-creased,while the expression of Cyclin D1,MMP-2,MMP-9 and phosphorylation of AKT were obviously reduced,with statisti-cal significance (P<0.05). Compared with single drug group,the effect of drug combination group strengthened,with statistical significance in above indicators(P<0.05). CONCLUSIONS:The inhibition effect of paclitaxel combined with cisplatin on the pro-liferation,migration and invasion of thyroid cancer cells SW579 cell will be strengthened,by a mechanism of up-regulating the ex-pression of PTEN and p27,down-regulating the expression of Cyclin D1,MMP-2 and MMP-9,inhibiting phosphorylation of AKT.

Objective To study effects of polo-like kinase-1(PLK1)small interfering RNA(siRNA)on proliferation and apoptosis of undifferentiated human thyroid cancer cells.Methods 5 PLK1 siRNA(S1,S2,S3,S4 and S5)were constructed and used to transfect human thyroid cancer cell line ARO.RT-PCR was employed to pick out the most effective siRNA,which was then used to transfect ARO cell.RT-PCR and western blot were used to detect PLK1 expression in thyroid cancer cells,which were divided into different groups.MTT assay was performed to examine the effects of PLK1 siRNA on thyroid cancer cells in all groups.Apoptosis of thyroid cancer cells was observed by caspase-3 activity and TUNEL.Results All the 5 siRNA down-regulated PLK1 mRNA expression.among which S4 showed the best effect.S4 transfection could obviously inhibit proliferation of thyroid cancer cells in dose and time dependent manner.Compared with control groups,caspase-3 activity of cancer cells in s4 transfeeted group increased significantly.The effect of S4 transfection was dose and time dependent.TUNEL results showed apoptosis of cancer cells transfected by S4 siRNA was obvious and apoptosis of cells was dose-dependent.Conclusions PLK1 may play an important role in proliferation of undifferentiated thyroid carcinoma.PLK1 siRNA transfection can inhibit proliferation of throid cancer cell through apoptosis induction.

Objective To study the effects of midkine(MK)gene small interfering RNA(siRNA)on adhesion and invasion of human breast cancer cells.Methods Real time PCR was used to evaluate MK mRNA expression in 7 human breast cancer cell lines Bcap-37,LCCI,MCF-7,MDA-MB-231,MDA-MB-435,MDA-MB-468,and ZR75-1.The cell line in which MK expression was the highest was transfected with different doses of MK siRNA.The expression of MK mRNA and protein was determined by real-time quantitative PCR and immunoflurescence staining.The cell adhesion was evaluated by MTT assay and invasion was examined by Boyden chamber method.Results Cell line MCF-7 expressed the highestlevel of MK mRNA in the 7 tested breast cancer cell lines.After being transfected with MK siRNA,MK mRNA and protein level of MCF-7 decreased in timeand dose-dependent manners.The adhesive and invasive ability of MCF-7 cell transfected with MK siRNA decreased in a dose dependent manner(P<0.01,P<0.01).Conclusions MK gene might play an important role in adhesion and invasion of human breast cancer cells.siRNA transfection could effectively inhibit adhesion,migration,and invasion of human breast cancer cell.

Objective To investigate the effects of S100A6 gene on invasion of human pancreatic cancer cell and possible mechanism. Methods Human pancreatic cancer BxPC3 cell line was transfected with small interfering RNA (siRNA) targeting S1006 gene, the mRNA and protein levels of S100A6 were determined by real time RT-PCR and Western blotting respectively. The invasion ability was evaluated by Transwell chamber. The matrix metalloproteinase-2 (MMP-9) activity of cancer cells was examined by gelatin zymography. Results The levels of mRNA and protein of S100A6 were greatly reduced in a dose and time dependent manner, the number of penetrating cells was greatly reduced in a dose dependent manner. The expression of S100A6 mRNA in 12.5 nmol/L of S100A6 siRNA transfected group decreased from ( 100 ±0.3)％ in control group to (15.3 ±0.2)％ ; while the expression of S100A6 protein decreased from (83.2 ±0. 18 ) ％ to ( 13.5 ± 0. 12) ％ ; the number of penetrating cells decreased from 44.5 ± 2.2 to 7.6 + 1.5 ( P ＜0. 01 ). The MMP-9 activity of siRNA group reduced significantly. Conclusions S100A6 siRNA can inhibit the invasion of pancreatic cancer cells through down-regulation of MMP-9.

ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.