Objective To investigate the effect of S100A4 silencing on tumor related gene COX-2,bcl-2,Surviving,MMP-9 mRNA expressions of pancreatic cancer BxPC-3,AsPC-1 cells,and explore their relationship.Methods Small interfering RNA interfering S100A4 gene (siRNA-S100A4) was applied to transfect human pancreatic cancer BxPC-3,AsPC-1 cells,and nonhomologous siRNA-C was used as negative control,and cells without transfection were used as control group.The expressions of S100A4,COX-2,Survivin,MMP-9,bcl-2 mRNA after interference were detected by using RT-PCR.Results S100A mRNA expressions of BxPC-3's control group,siRNA-C group,siRNA-S100A4 group were 0.661 ± 0.023,0.659 ± 0.043,0.379 ± 0.039,and expressions of COX-2 mRNA were 0.760 ± 0.026,0.830 ± 0.017,0.443 ±0.006,and expressions of Survivin mRNA were 0.948 ± 0.049,0.909± 0.081,0.068 ± 0.006,and expressions of bcl-2 mRNA were 0.462 ±0.018,0.421 ±0.049,0.184 ±0.025,and expressions of MMP-9 mRNA were 0.813 ± 0.008,0.908 ± 0.063,0.246 ± 0.027.S100A mRNA expressions of AsPC-I's control group,siRNA-C group,siRNA-S100A4 group were 0.641 ± 0.042,0.626-± 0.053,0.320 ± 0.081,and expressions of COX-2 mRNA were 0.727 ± 0.021,0.743 ± 0.025,0.560 ± 0.035,and expressions of Survivin mRNA were 0.994 ± 0.032,0.984 ± 0.049,0.063 ± 0.005,and expressions of bcl-2 mRNA were 0.458 ±0.004,0.537 ± 0.046,0.181 ± 0.007; and expressions of MMP-9 mRNA were 0.698 ± 0.011,0.718 ± 0.073,0.199± 0.013.The expressions of S100A,COX-2,Survivin,bcl-2,MMP-9 mRNA in groups with siRNA-S100A4 transfection were significantly lower than those of siRNA-C group and control group (P ＜0.01),but the difference between siRNA-C group and control group was not statistically significant.Conclusions S100A4 plays a role in the pathogenesis of pancreatic cancer through up-regulation of COX-2,Survivin,bcl-2,MMP-9 expressions.

Objective To provide theoretical and scientific evidences for plague control,through understanding the F1 antibody level distribution and affected factor of population having healed from plague from plague natural focus of Rat.flavipectus in Yunnan Province.Methods The places and population investigated were chosen according to plague surveillant data in Yunnan Province from 1986 to 2005,using caso-control study and quesfionary. All samples were detected by indirect hemagghtination(IHA),including 248 serum samples from population having heaIed from phsue in 23 counties as case group.295 senlm samples from healthy population inoculated with EV vaccine in 7 counties as artificial immunization group, and 235 serum samples from healthy population not inoculated it in a non-plagued foei county as negative comparison group,with the diagnosing standard for positive titor being not less than 1:20.Results(①The difference WSS statistically significant(X2=44.80,P<0.05)between plague and non-plagued foci with F1 antibody positive rates being 22.10%(120/543)and 0(0/235),respectively.② The F1 antibodv positive rate of case group,35.89%(89/248)and geometric mean titer(GMT)1:84,was higher than that of artiIicial immunization group,which was 10.51%(31/295)and with GMT 1:34,respectively,the difference being statistically significant(X2=50.41,P<0.0125);the positive rate of case group wgs hisher than the neganve comparison group,the difference being statistically significant(X2=103.39,P<0.0125):the posifive mte of artificial immunization group was higher than the negative comparison group,the difference being statisticallv significant(X2=26.23,P<0.0125).③The differences were not statistically significant in the F1 antibedy positive rates of case group for age,sex,nation and occupation(X2=1.88,2.01,5.46,0.04,P>0.05).④The difference was not statistically significant in 89 plague patients with positive F1 antibody at the time of onset and rehabilitation(t= 1.23,P>0.05).Conclusions ①Plague FI antibody in people distributes the sanle a8 the plague natural focus of Rat.flavipectus does in Yunnan Province.②For naturally infected plague patients,only 1/3 popuhtion get long- term immunity,and still 2/3 can be infected again.The protecting rate and effect of naturally acquired immunity due to infection of plague are better than amfieially acquired immunity from inoculation of EV vaccine.③For the population having healed from plague,the positive rotes of FI antibody are not affected by age,sex,nation and occupation,however for those whose plague F1 antibody is still positive after some time,the titer will remain or even increase.

Objective To investigate the alternation of insulin -like growth factor -Ⅱ( IGF-Ⅱ) gene promoter P4 methylation status in hepatocellular carcinoma ( HCC) and explore its relationship with expression of P4 mRNA levels.Methods Liver specimens of 43 patients with HCC and normal liver specimens of 9 control patients were collected in operation .Tissue DNA and total RNA were extracted from these specimens .IGF-Ⅱ P4 methylation status and P4 mRNA expression levels were detected .Results (1)The incidence of IGF -ⅡP4 methyl-ation in HCC group was significantly lower than that in normal liver specimens (16.28%vs 88.89%,χ2 =19.12,P<0.01).(2)The expression level of IGF -ⅡP4 mRNA in HCC group was significantly higher than that that in normal liver specimens[(0.96 ±0.74) vs (0.25 ±0.19),t=5.48,P<0.01].(3)In HCC group,the IGF-Ⅱ P4 mRNA expression level with hypomethylation gene was significantly higher than that without hypomethylation gene [(1.18 ± 0.76) vs (0.32 ±0.27),t=5.28,P<0.01].Conclusion The hypomethylation alternation of IGF -Ⅱ P4 gene promoter which is accomplished by up -regulate P4 mRNA expression has a close relationship with HCC .

BACKGROUND: There are many drawbacks with hormone replacement therapy for menopausal syndrome. The blood levels of 5-HT and norepinephrine are lower. Fluoxetine hydrochlorde(ProzacR) is a selective serotonin reuptake inhibitor which is widely used in treating anxiety and depression,OBJECTIVE: To evaluate the effect of combined antidepressant and estrogen therapy compared to estrogen alone in the treatment of perimenopausal syndrome in this prospective open study.DESIGN: Randomized comparative study.SETTING: Department of Obstetrics and Gynecology of the First Affiliated Hospital of Chongqing Medical University PARTICIPANTS: From November 2003 to December 2004, 60 female patients with diagnosed menopausal syndrome of 3-12 month duration, aged (46±3) years, from Department of Obstetrics and Gynecology of the First Affiliated Hospital of Chongqing Medical University were enrolled into the study after giving their informed consents. The patients were randomly divided into two equal groups with Group 1 (n=30) receiving a combination of antidepressant + estrogen and Group 2 (n=30) receiving estrogen only.METHODS: Patients in Grgup 1 received fluoxetine hydrochloride (ProzacR) 20 mg orally every morning plus oral estradiol 1 mg once every two weeks for 2 months. Patients in Group 2 received only oral estradiol 1 mg once every two weeks for two months. Patients were not taking any other drug during the treatment period. At the end of two month treatment all patients were evaluated with the following 3 assessment tools: ①female menopausal symptom evaluation with the following 4 categories: Complete symptom relief, markedly improved, improved and no effect. Overall efficacy was defined as 50% symptom improvement. ② Hamilton Depression Scale which reflects energy level and psychosomatic factor of sleep and anxiety. ③Menopause index: Which are description of clinical evaluation and adverse effects; this would help to assess the safety of using both drugs in treating the menopausal syndrome.MAIN OUTCOME MEASURES: female menopausal symptom assessment, hamilton depression scale, and menopause index.RESULTS: ① In female menopausal symptom assessment group 1 showed better results in the complete relief and markedly improved scores. ②Hamilton Depression Scale group 1 also showed better scores than Group2(In Groupl, the scores at week 1 to 8 were 25,18,15,10,8,5,5,4 respectively ,in Group 2, the scores at week 1 to 8 were 25,17,15,14,13,12,13,13 respectively). ③ Group 3 showed a significant better score in the menopausal index with improvement in sleep disorder, anxiety and depression than Group 2 (In Group 1 the scores at week 1-8 were 32,22.5,15,15,14,15,15,14 respectively,In group 2, the scores at week 1 to 8were 33,21,16,14,13,12,13,13 respectively) ,there was no significant difference of incidence of adverse events as compared with Group 3 .CONCLUSION: Combined therapy of fluoxetire hydrochlarde(PROZAC)plus estrogen showed better efficacy in the treatment of menopausal syndrome than estrogen alone.

Objective To analyze the thin-section CT features of fissure-attached nodules in asymptomatic patients.Methods The baseline and follow-up CT images of 120 asymptomatic patients with fissure-attached nodules were retrospectively analyzed.The thin-section CT findings include size,shape,density,interface,and the relationship with pulmonary fissure.Results There were 1 67 fissure-attached nodules of the 120 patients.The mean length of nodules was 5.46 mm±2.276 mm (2 - 1 5 mm).104(104/1 67,61.1%)nodules were less than 5 mm in length.1 63 (1 63/1 67,97.6%)nodules were typical and 4 (4/1 67,2.4%)nodules were atypical.All these nodules manifested as homogenous,solid,smooth interface,and widely adjacent to the fissure on CT ima-ges.Among those nodules,1 64(1 64/1 67,98.2%)nodules were triangular(88/1 67,52.7%)or oval(76/1 67,45.5%).126(126/1 67, 75.4%)nodules were abutting the major fissure.The angle between nodule and fissure was obtuse in 1 64(1 64/1 67,98.2%)fissure-attached nodules.During follow-up,4(4/1 67,2.4%)nodules increased in size and the corresponding volume doubling time (VDT) range was 863-4 228 days.However,no obvious malignant signs were found.Conclusion The thin-section CT images of fissure-attached nodules in asymptomatic patients showed the benign characteristics,most of which less than 5 mm and demonstrated trian-gle or oval,smooth interface and homogenous solid density.

Objective To evaluate the correlation between thin-section CT morphologic features and pathological vascular invasion in the clinical stage ⅠA peripheral lung adenocarcinoma.Methods The preoperative thin-section CT images,clinical and pathological characteristics of 102 nodules in 98 patients were retrospectively analyzed and divided into two groups according to pathological vascular invasion.Variables including the diameter of tumor(T) and consolidation part (C) in the mix ground glass nodule (mGGN),C/T ratio,morphology and pathological classification were analyzed by two-independent samples Mann-Whitney U test or t test and x2 test or Fisher exact probability test.Variables with significant difference in the univariate analyses were entered into multivariate analysis to explore predictors for vascular invasion.The correlation between these measurements and vascular invasion was evaluated by a ROC analysis.Results All of 102 nodules,36 had pathological vascular invasion and 66 without.The numbers of two groups with speculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation were 30,30;26,26;35,3;35,39;32,39 and the mediandiameter of tumors were 2.1(1.0-3.0),1.2(0.5-3.0) cm respectively.By univariateanalysis,spiculation,air bronchogram,abnormal vein,vascular convergence,pleural indentation,and diameter of tumor were significantly associated with pathological vascularinvasion (P＜0.01).By multivariate logistic analysis,the statistically significant difference were found in speculation and abnormal vein (OR=33.867,8 161.063,P＜0.05).The ROC analysis showed the area under curve,cut-off value,sensitivity and specificity ofthe diameter of tumor and consolidation part of mGGN and C/T ratio were 0.784,2.05 cm,66.7％,78.8％;0.886,1.08 cm,82.4％,90.0％ and 0.861,0.65,79.4％,85.0％ respectively in differentiating tumors with pathological vascular invasion.Conclusion The spiculation sign and abnormal vein are independent predictors for vascular invasion in stage ⅠA peripheral lung adenocarcinoma.

OBJECTIVE: To explore a research path and statistical analytical method for cost-utility analysis suitable for gallstone surgery patients.METHODS: A decision-making model for the treatment of cholelithiasis surgery was established;the costs(C) and utility values at each nodal point(? U) were collected,with the results subjected to sensitivity analysis and threshold analysis.RESULTS: The cost-utility ratios(CUR) of laparoscopic operation and open surgery were 4 605.6(? 30%) yuan/QALY and 5 979.7(? 30%) yuan/QALY,respectively.One dimension sensitivity analysis of each( every) parameter reveals that the former CUR value is greater than the latter one.C1 and ? U1 showed the biggest impact on the CUR of laparoscopic operation,while C5 and ? U5 did on the CUR of open surgery.When C1=10 205.6 yuan,or ? U1=1.303 or C5=10 979.8 yuan or ? U5=3.095,CUR of laparoscopic operation was equal to CUR of open surgery,which means that the conversion of threshold was achieved.CONCLUSION: The designed research path and statistic analytic methods are suitable for the cost-utility analysis of cholelithiasis surgery patients.

Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

Air ; analysis ; Chromatography, Gas ; Propane ; analogs & derivatives ; analysis ; Reproducibility of Results ; Workplace

Objective To construct the RNA interference eukaryotic expression vector targeting human centromere protein-I (CENP-I) and to observe its effect on the growth of human embryo kidney 293 cells (HEK 293). Methods The expression vectors of pGenesil-1/CENP-I-siRNA-1. pGenesil-1/CENP-I-siRNA-2 and pGenesil-1/CENP-I-siRNA-3 were constructed by gene recombination and then were transfected into the HEK293 cells by liposome. The expressions of CENP-I at the protein and mRNA levels were detected by Western blotting and fluorescence quality PCR (FQ-PCR). The effective vector and the best transfection time were selected. The growth and the cell cycle of the transfected cells were assessed by MTT assay and flow cytometry. Giemsa was used to stain the transfected cells to calculate the mitotic index. Results Sequence-specific siRNAs targeting CENP-I significantly down-regulated the expression of CENP-I in HEK293 cells. The recombinant plasmid of pGenesil-1/CENP-I-siRNA-3 was the effective vector. After transfecting for 72 h the best inhibited efficiency was achieved. In CENP-I-siRNA transfected cells,the rate of cell growth was decreased markedly. Cells at G 2/M phase and the mitotic index were increased conspicuous compared with the cells transfected with the blank vector or untransfected. Conclusion Down-regulation of CENP-I in HEK293 cells by sequence specific siRNA delays the cell growth and postpones the cell division.