1.Operating principle and structure of automatic system in laboratory
Wencai LAN ; Feng LAN ; Di PAN
Chinese Medical Equipment Journal 1993;0(05):-
This article introduces the principle and the structure of streamline testing based on the newly developed laboratory automation system in our hospital.An overview of the functions and the technical structure of each module within streamline testing are provided.The streamline procedure and the information transmission between modules are also briefly described.
2.Suppression effect of RNA interference on expression of EGFR of SKOV3 cell line
lan, DAI ; wen, DI ; chuan-wei, DING
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(01):-
Objective To investigate the suppression effect of RNA interference(RNAi) on the expression of epidermal growth factor receptor(EGFR) of ovarian cancer SKOV3 cell line. Methods SKOV3 cells were transfected with synthetic EGFR sequence-specific dsRNA by lipofectamin 2000.The expression level of EGFR mRNA and protein were detected by RT-PCR and Western blotting,respectively.The proliferation of transfected SKOV3 cells was detected by WST-1 cell proliferation assay.The abilities of adhesion,migration and invasion were detected by cell adhesion assay,cell migration assay and Matrigel invasion assay,respectively. Results The suppression rates on EGFR mRNA and protein by sequence-specific dsRNA-EGFR were 73.65% and 58.94%,respectively.The cell proliferation was inhibited by 22.54%,35.76% and 31.07%,respectively at 24,48 and 72 h after transfection.The adhesion of sequence-specific dsRNA transfected SKOV3 cells for 30 min and 90 min were reduced by 12.11% and 18.66%,respectively.The migration and invasion of SKOV3 cells were decreased by 25.47% and 22.08%,respectively. Conclusion The down-regulation of EGFR expression of ovarian cancer by sequence-specific dsRNA can lead to the suppression of proliferation,adhesion,migration and invasion of ovarian cancer cells.
3.Occupational security awareness and occupational protection skills among community nurses
Lan-Di WU ; Wei-Li HAN ;
Chinese Journal of General Practitioners 2005;0(12):-
Objective This investigation was to define the occupational security-related factors in community nurses and effective strategies to improve their service awareness and occupational protection ability.Methods A questionnaire was used for 1702 community nurses in Guangzhou to learn occupational security-related factors and occupational protection skills.Results Self-report results found that 21.3% (362/1702) nurses fully understood the occupational security-related factors,12.0% (204/1702) nurses knew the requirement for occupational security,and 12.3% (209/1702) nurse could provide occupational protection.Conclusions These results suggested that most community nurses showed poorer occupational protection awareness and ability.Thus,occupation safety management regulations,standardized performance procedure and emergency response plan should be put forward,and further community nurse training programme need to be initiated.
4.Ulinastatin attenuates paraquat-induced myocardial injury
Li LI ; Min DI ; Chao LAN ; Changhua SUN
Chinese Journal of Emergency Medicine 2012;21(11):1192-1197
Objective To investigate the effect of ulinastatin (UTI) on endogenous antioxidant systems in the process of myocardial oxidative stress injury induced by paraquat (PQ),so as to elucidate UTI protecting cadiocytes against PQ-induced injury.Methods Thirty Japan white rabbits were randomly (random number) divided into A,a sham control group; B,PQ intoxication model group and C,D,E,three UTI groups as per different dosages of UTI given.PQ (30 mg/kg) was administered by intraperitoneal route to the rabbits of PQ intoxication model and rabbits of three UTI groups.UTI in doses of 15 000,30 000,50 000 U/kg was administered intravenously every day to the rabbits of three UTI groups respectively for 7 days after PQ intoxication.After the rabbits were sacrificed with 10% chloral hydrate,left ventricles of rabbits were taken.Histomorphological changes of myocardium were observed by using HE staining.Hydroxyproline was measured by alkaline hydrolysis method,myocardial oxidative stress evaluated by 4-hydroxy-2-nonenal (4-HNE) immunohistochemistry staining,transcription levels of transforming growth factor-β1 (TGF-β1),adiponectin,peroxisome proliferator-activated receptor α (PPAR-α),AMP-activated protein kinase-β1 (AMPK-β1),uncoupling protein-1 (UCP-1) were assayed with reverse transcriptionpolymerase chain reaction (RT-PCR).The differences between two groups were analyzed by t test ; for multiple comparisons,data were analyzed by One-Way ANOVA followed by Dunnett' s post hoc test; for two and multiple ranked data comparisons,Mann-Whitney U test and Kruskal-Wallis test were used,and Spearman rank correlation analysis was used to investigate the correlation between UTI dose and transcription levels of endogenous antioxidant.Results Compared with group A,left ventricular cadiocytes in group B became swelled and disarranged,concentration of tissue HYP was elevated to (2.37 ± 0.49),P =0.001;scores of 4-HNE and TGF-β1 mRNA expressions were all upregulated (both P =0.001).Compared with group B,disordered myocytes in UTI treated groups tended towards normal,4-HNE scores in group D and group E descended respectively to (1.83 ± 0.53) and (1.70 ± 0.47),both P =0.001,HYP concentration reduced to (3.51 ±0.39) μg/mg and (3.29 ±0.37) μg/mg (P =0.002; P =0.001),and expressions of TGF-β1 mRNA in these two groups were decreased (both P =0.001).Transcription levels of four endogenuous antioxidant components,adiponectin,PPAR-α,AMPK-β1 and UCP-1,were all augmented in the group E with high dose UTI.The mRNA expressions of these four components positively correlated with the dose of UTI in the myocardium of PQ intoxicated rabbits.Conclusions UTI could protect myocardia against PQ-induced injury by increasing endogenous antioxidant systems.
5.Optimization of radiological protection of patients undergoing digital radiography
Menglong ZHANG ; Yuting DUAN ; Yujun XU ; Di LAN ; Liangan ZHANG
Chinese Journal of Radiological Medicine and Protection 2013;(2):183-186
Objective To identify the necessity to carry out optimization procedure in routine digital radiography (DR) by evaluating changes of patient radiation dose and image waste ratio before and after optimization.Methods Two hundred patients with near-standard body build were enrolled in the study.Half of them undertook routine examination,and the others undertook the examination with bestlyoptimized protocol.The dose-area product (DAP) and entrance surface dose (ESD) were recorded.The image waste ratios in 2 groups were calculated and the reasons for image waste were analyzed.The radiation dose and image waste ratio before and after optimization were compared.Results The ESD,DAP and image waste ratio in bestly-optimized radiography were significantly lower than those in non-optimized radiography (z =9.31,16.22,P<0.05; x2 =36.5,P < 0.05).Conclusion Using the bestlyoptimized digital radiography,the patient radiation dose and image waste ratio are effectively reduced.
6.Researching on fingerprint of Inulacappa by HPLC.
Di-fei-fei XIONG ; Di ZHU ; Dan TAN ; Yan-yu LAN ; Lin ZHENG ; Ai-min WANG
China Journal of Chinese Materia Medica 2015;40(3):480-483
OBJECTIVEThis study is to establish the fingerprint and find out the common chromatographic peaks of Inula cappa by HPLC.
METHODThe HPLC analysis was performed on an Agilent Eclipse Plus C18 column (2.1 mm x 150 mm, 1.8 μm) with 0.1% fomic acid aqueous solution-0.1% fomic acid acetonitrile solution as mobile phase at a flow rate of 0.3 · mL(-1) · min(-1); The detective wavelength is 325 nm; The column temperature is 45 °C.
RESULTThe results indicated that 5 of 17 common peaks were identified . The similarity about 10 groups of Inulacappais is over 0.95.
CONCLUSIONThis method is able to be a scientific basis of quality assessment according to its convenient and reliable.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Inula ; chemistry
7.The expression and clinical significance among CD44 +/CD24 -,E-cadherin and ivm entin in non-sma ll cell lung canc er
Xueqing GUAN ; Leiguang YE ; Yue GAO ; Xun QIU ; Lan WANG ; Di WANG ; Baogang LIU
Practical Oncology Journal 2014;(4):315-320
Objective To detect the expression of CD 44 +/CD24 -and E-cadherin and vimentin in non-small cell lung cancer and its clinical prognostic relevance .Methods By immunohistochemistry , detecting , 500 cases of non-small cell lung cancer E -cadherin ,Vimentin and CD44 +/CD24 -expressions′correlation ,we analyzed the relationship with non -small cell lung cancer .Results In non-small cell lung cancer , vimentin was related to tissue differentiation,TNM staging(P<0.05);there was relationship between CD44 +/CD24 -and EMT by E-cadherin expression(P <0.05);CD44 +/CD24 -expression was related to lymph node metastasis , differentiation(P<0.05);their expressions were not obviously related to age ,sex,smoking history,clinical patho-logical type .The survival time of CD44 +/CD24 -high expression in NSCLC was significantly shorter than lower expression in NSCLC(P<0.05).Conclusion EMT malignant phenomenon raises cancer stem cells ,which can lead to the development of malignant cells poorly differentiated trends ,they are not an isolated process;important role in the development of EMT and CD 44 +/CD24 -union occurs in non -small cell lung cancer and prognosis with patients ,which provides a potential breakthrough in the treatment of cancer prevention and treatment targets .
8.Correlation of LIPC C480T (rs1800588) polymorphism with lipid metabolism in Han Chinese population
Xiaohua TAN ; Chunhong DI ; Baodong WANG ; Qiang MENG ; Xianhong HUANG ; Man LAN ; Lei YANG
Chinese Journal of Laboratory Medicine 2012;35(10):905-911
Objective To investigate the association of the LIPC-C480T (rs1800588) and lipid levels and dyslipidemia in different age-and-sex groups in Han Chinese population.Methods The serum TC,TG,LDL-C and HDL-C were detected by automatic biochemical analyzer in 2420 health adults (1527 men and 893 women).The genotypes of rs1800588 were detected by M ALDI-TOF MS.According to the age difference (≤44,45-59 and ≥60-year-old),the total samples were divided to young (241 men and 201 women),middle-aged (652 men and 360 women) and older (634 men and 332 women) groups.The effects of genotypes on 4 serum lipid indicators in each age-and-gender group were analyzed by one way analysis of variance (ANOVA),and the odd risk of genotypes on dyslipidemia was estimated by binary Logistic regression analysis.The P value less than 0.05 was considered statistically significant.Results The frequence of allele T for LIPC rs1800588 in this population is 39.4%.In each age group the lipid parameters are quite different between males and females.Compared with those with CC genotype,middle-aged and elder men with CT or TT genotype have higher TC and HDL-C levels,and elder men with TT genotype also have higher TC level ; young women bearing CT genotype have higher TC level,and the CT and TT genotypes have higher HDL-C levels,middle aged women with CT or TT genotype have higher TC and TG levels,and CT genotype also have higher HDL-C level,the elder women with TT genotype have higher HDL-C level.Compared with those CC genotype individuals,the risk for mixed hyperlipidemia and hypercholesterolemia increases 2.318 folds (P =0.004) and 2.571 folds (P < 0.001) respectively,while the risk for low HDL-C decreases 1.908 folds (P =0.029) for TT genotypes individuals among elder males; the hypercholesterolemia risk increasc 1.688 (P =0.036) and 2.099 times (P =0.040) in CT and TT genotypes respectively,and the risks for hypertriglyceridemia and mixed hyperlipidemia are 2.060 (P =0.038) and 2.381 (P =0.019) times higher than those with CC genotype among middle-aged females.Conclusions The LIPC rs1800588 site associates with the lipid levels and dyslipidmia risk in Han Chinese in an age-and-sex model.This SNP site has higher impact on lipid levels and dyslipidemia among elder males and middle-aged females,and the T allele is the risk factor.
9.Regulation of microRNA-199a on adhesion, migration and invasion ability of human endometrial stromal cells
Lan DAI ; Liying GU ; Jie ZHU ; Jun SHI ; Yao WANG ; Fang JI ; Wen DI
Chinese Journal of Obstetrics and Gynecology 2011;46(11):817-821
Objective To study the regulation of microRNA 199a (miR-199a) on adhesion,migration and invasion ability of human eutopic endometrial stromal cells (ESC) from patients with endometriosis.Methods ESC were transfected with miR-199a mimics or negative control (NC) RNA by lipofectamine 2000.The adhesion,migration and invasion ability of ESC were detected by cell adhesion assay,scratch assay,cell migration assay and matrigel invasion assay,respectively.Luciferase reporter assay was used to evaluate whether IKKβ was the target gene of miR-199a.The expression of ikappa B kinase beta (IKKβ),inhibitory kappa B alpha (IκB-α),phospho-IκB-α (p-IκB-α) and nuclear factor-kappa B (NF κB) protein were measured by western blot.Results ( 1 ) Adhesion potential:the adhesion inhibitory rates were ( 14 ± 4 )% in miR-199a group and 0 in control group,which showed significant difference (P<0.01 ).(2) Migration and invasion:in the scratch assay,ESC transfected with miR-199a exhibited a lower scratch closure rate than that of controls.In migration and invasion assays,the migration and invasion ability of miR-199a group were significantly decreased compared with those of NC group [ 130 ± 31 vs.247±36 (P<0.01); 63 ± 15 vs.133 ± 17 (P<0.01),respectively].(3) The luciferase activity of miR-199a group was significantly lowered than that of control group [ 0.160 ± 0.006 vs.0.383 ± 0.083 ( P <0.01 ) ].The protein levels of IKKβ,p-IκB-α,IκB-α and NF-κB of 0.350 ±0.195,0.443 ±0.076,1.970 ±0.486 and 0.454 ± 0.147 in miR-199a group were significantly different compared with the NC group in which the protein levels were set at 1.000 ( P < 0.01 ).Conclusions miR-199a can inhibit the adhesion,migration and invasion of the ESC.IKKβ is the target gene of miR-199a in ESC.One of the mechanisms of the inhibition effect is probably that miR-199a inhibits the activation of NF-κB signaling pathway by targeting IKKβ gene.
10.The effects of hydrogen sulfide on expression of HIF-1α mRNA in myocardium of rats after cardiopulmonary resuscitation
Di CHEN ; Hao PAN ; Chunwen LI ; Xiucai LAN ; Beibei LIU ; Guangtian YANG
Chinese Journal of Emergency Medicine 2011;20(7):687-691
Objective To investigate the changes of the expression of HIF-1α (hypoxia induciblefactor-1a) mRNA in myocardium of rats after cardiopulmonary resuscitation (CPR) and the intervention effects of exogenous hydrogen sulfide on it. Method Forty five male SD rats were randomly divided into three groups: control group ( n = 15 ), cardiopulmonary resuscitation group ( CPR group, n = 15 ) and NaHS + CPR group (n = 15 ) . Rats of control group were anesthetized and intubated without asphyxia and cardiac arrest. The rats of CPR group and NaHS + CPR group were operated to induce cardiac arrest by asphyxiation. In the rats of NaHS + CPR group, NaHS in dose of 50 ug/kg was administrated via the femoral venous line 1 minute before CPR. Hemodynamic was monitored continuously. The expression of HIF - lα mRNA in myocardium of rats in each group was determined by using RT-PCR and the activity of myeloperoxidase (MPO) in the myocardium of rats in each group was assayed by using a patent reagent box 6 h after CPR. The histopathological changes of myocardium were also observed. The t- test was used for statistical analysis. Results There was no statistically significant difference in hemodynamic changes between CPR group and CPR + NaHS group ( P > 0. 05 ) . When compared with the control group, the activity of MPO and the expression of HIF-1α mRNA in CPR group and CPR + NaHS group were both increased, and those increased in CPR + NaHS group was more significant (P < 0. 05) . When compared with CPR group, the expression of HIF-1α mRNA in CPR + NaHS group was higher, however, the activity of MPO in CPR + NaHS group was lower ( P < 0. 05) . There were various histopathological changes of myocardium of rats found in CPR group and CPR + NaHS group, and the damage of myocardium of rats in CPR group was more obvious than that in CPR + NaHS group. Conclusions The expression of HIF-1α mRNA in myocardium of rats was increased after CPR. Exogenous hydrogen sulfide can protect myocardium cells from resuscitation-perfusion injury, and the protection is associated with increase in expression of HIF 1 αmRNA.