Purpose To investigate the expression of Netrin-1 in ovarian serous carcinoma and its c1inicopatho1ogica1 significance. Methods Netrin-1 protein expression was assessed by immunohistochemica1 method in tissue specimens from 20 cases of benign ovari-an serous cystadenomas,13 cases of border1ine ovarian serous neop1asms and 32 cases of ovarian serous carcinomas( OSC). Results The positive proportion of Netrin-1 protein in OSC tissues was significant1y higher than those in border1ine and benign ovarian serous ne-op1asms(P<0. 01). The expression of Netrin-1 in OSC tissues was associated with tumor grade and c1inica1 stage(P<0. 05),but not associated with age,site,tumor size or 1ymph node metastasis( P>0. 05). Kap1an-Meier ana1ysis showed that the 5-year surviva1 rate of patients with Netrin-1 over-expression was significant1y 1ower than that of patients with 1ower expression( P<0. 05 ). Conclu-sions The high expression of Netrin-1 in OSC tissues indicates that Netrin-1 p1ays an important ro1e in cancer pathogenesis and deve1-opment and it may be a new assistant marker for prognosis of OSC.

Objective To investigate the role of nuclear factor-kappa B(NF-?B)/inhibitor protein-kappa B(I?B) signal pathway in viral transactivation of transcription in steroid responsive simple nephrotic syndrome(SRSNS).Methods Children with SRSNS(inclu-ding active stage and remissive stage) were examined,and were compared to children with nephritic nephrosis,secondary glomerular di-seases,bronchiolitis and healthy children.Electro-mobility shift assays,reverse transcription-polymerase chain reaction(RT-PCR) and enzyme-linked immunosorbent assay(ELISA) were used to detect the activity of NF-?B,the gene expression of respiratory tract viruses (including respiratory syncytial virus and influenza virus) in peripheral blood mononuclear cells(PBMCs) and the levels of viral antibody in plasma,respectively.The protein levels of I?B? and IL-8 were measured through Western blot and ELISA in SRSNS at active stage and healthy children.Results Compared with SRSNS at remissive stage and other groups,the activity of NF?B in SRSNS at active stage was much higher.And there was a positive linear correlation trend between the activity of NF-?B and the gene expression of respiratory tract viruses in SRSNS at active stage.With healthy children,the level of IL-8 in plasma from SRSNS at active stage was significantly increased.There was a positive correlation between the activity of NF?B and the level of IL-8(r=0.88 P

Objective To explore the value of manganese-enhanced MRI in locating the rat visual nuclei.Methods The visual nuclei of thirty-six rats were located by 3 different ways including individual MEMRI locating (group A,n= 1 6),anatomical atlas locating (group B,n=1 6)and direct puncture by using the data obtained in MEMRI (group C,n=4).After unilateral intra-vitreal injection of MnCL2 (30 mmol/L×3 μL)in group A,the brain MRI was performed 24 h later.The location coordinate of lateral geniculate nucleus (LGN) and superior colliculus (SC)were recorded individually.The nuclei injections (3% fluorogold solution,1 μL)were performed by using different location coordinate in groups A and B.The rat’s retinas were examined under fluorescence microscope 5 days later,and the results were compared between the two groups.After brain nucleus puncture injection (30 mmol/L MnCL2 solution,0.5 μL),MRI was performed 1 h later in group C.Results The success rate was 93.8% (1 5/1 6)in group A,and 65.5% (10/1 6)in group B.The difference between groups was statistically significant (P<0.05).All the injection locations of C group were agreed with atlas.Conclusion MEMRI in the visual nucleus stereotactic can improve the accuracy of location.

Objective To assess the advantages of multi-source RF transmission(MT) for balanced fast field echo(BFFE) cardiac cine imaging.Methods Fifteen volunteers were scanned on a clinical 3.0 T MR system equipped with MT.Acquisition of B1 maps with and without MT were followed by axial and four chambers BFFE cine imaging for all subjects with different transmission modes[single source RF transmission (ST),MT and MT with allowable shortest TR(MTS)].The B1 field uniformity and contrast to noise ratio (CNR) were quantitatively analyzed.The B1 field uniformity was evaluated by the pixel values within the ROI and was compared with Student t test.CNR was defined as follows:(SIblood-SImyocardium)/ 0.5 ×(SDblood + SDmyocardium).CNR were tested with one-way ANOVA for three groups comparision and with LSD for inter-group comparison.Image qualities were blindly assessed by 2 readers with a 4-score scale.Global image quality scores were tested for statistical significance by using the nonparametric paired sample Friedman rank test for three groups and the Wilcoxon signed rank test for further inter-group comparison.The Kappa statistics was used to assess interreader agreement.Results B1 homogeneity was significantly improved for images with MT compared with ST under the condition of other parameters unchanged (t =21.632,P ＜0.01).In left ventricular,CNR of blood and myocardium was improved from 10.8 ±2.2 for ST to 14.4 ± 2.4 for MT and 14.1 ± 2.2 for MTS.In fight ventricular,it was improved from 7.7 ± 1.2 for ST to 12.2 ± 1.4 for MT and 12.0 ± 1.2 for MTS.The differences were statistically significant (F =11.617,61.472,P ＜0.01).The images of MT and MTS groups demonstrated greater CNR versus the ST images.There was no statistical difference between latter two groups.The image scores of Reader 1 and 2 were 4.60 ± 1.18 and 4.73 ± 1.10 for ST,6.53 ± 1.19 and 6.67 ± 1.29 for MT and 6.73 ± 1.03 and 6.73 ± 0.88 for MTS respectively.There were statistically differences among three groups (x2 =23.577,24.275,P ＜ 0.01).The image quality was improved on BFFE images using MT and MTS technology compared to ST technology,and there were also significant differences.The interreader agreement between two readers was good [K=0.643,0.722 and 0.814(P ＜0.05) for ST,MT,and MTS group respectively].Conclusions MT technology significantly improves B1 field uniformity,increases CNR and reduces artifact on BFFE cardiac cine images.The TR is decreased within the normal SAR ranges,and thus scanning speed is increased.

Objective To evaluate the application value of real-time three-dimensional echocardiography (RT-3D TEE) in nonvalvular atrial fibrillation patients after transcatheter left atrial appendage closure (LAAC) with the Amplatzer Cardiac Plug (ACP).Methods The two-dimensional transesophageal echocardiography (2D TEE) and RT-3D TEE were performed in selected patients to measure the diameter of left atrial appendage ostium and landing zone,to observe left atrial appendage morphology and lobular distribution.These were also performed to guide the whole process of transcatheter LAAC with ACP,which included the atrial septal puncture,sheathing canal cruise,occluder device implantation,and verifying the stability of occluder device and releasing the device.It involved observing ACP occluder morphology,location,stability,surrounding residual shunt,whether influencing the mitral valve and left superior pulmonary vein flow,and pericardial effusion.Results A total of 15 patients (100％) successfully underwent LAAC with the ACP.The maximum diameter of left atrial appendage ostium by 2D-TEE measurement during operation was (20.5 ± 2.9)mm,located at 135°.The diameter of left atrial appendage landing zone was (17.1 ± 2.8) mm,located at 45°;(18.0 ± 4.0) mm,located at 90°;(22.1 ± 4.7)mm,located at 135°,respectively.The left atrial appendage morphology:2 had one leaf and 13 had two leaves or more,including 2 cases of bifoliate short neck shape.In 15 patients,6 cases of cauliflower type,2 cases of wind sock type,3 cases of chicken wing type,2 cases of cactus type and 2 cases of complex type.The proximal left atrial appendage morphology:3 cases of boot type,2 cases of wide mouth type,6 cases of narrow mouth type,2 cases of straight tube type,and 2 cases of bifoliate short neck type.There was no obvious residual shunt in all the patients at immediately post-operation.Conclusions In the transcatheter LAAC with the ACP,RT-3D TEE has important application value in the preoperative selection of patients,the choice of appropriate occluder,guidance of full-process monitoring during operation,the postoperative effect evaluation and so on.

Objective To investigate the prevalence and molecular epidemiology of clinical isolates of plasmid-mediated 16S rRNA methylases-producing Klebsiella pneumoniae. Methods From January 2006 and September 2007, 337 non-replicate clinical isolates of Klebsiella pneumoniae were consecutively collected from inpatients in a teaching hospital in Wenzhou, China. All of the isolates were identified by the automated microbiology systems. MICs of amikacin, gentamicin and tobramycin were determined by agar dilution method. The isolates were investigated for the presence of ESBLs by the CLSI-recommended confirmatory tests. PCR was used to detect 16S rRNA methylase genes, ESBL genes and class Ⅰ integrase gene. The homology of these isolates was analyzed by pulse-field gel electrophoresis (PFGE). Results Sixty-four ( 19. 0% ), 28 ( 8. 3% ) and 55 ( 16. 3% ) of 337 isolates were resistant to gentamicin, amikacin and tobramycin, respectively. Twenty-one (6. 2% ) isolates carried 16S rRNA methylase genes (3 for armA, 13 for rmtB, 5 for both armA and rmtB) and high-level resistant to gentamicin, amikacin and tobramycin ( MICs ≥256 mg/L). Nineteen of 21 isolates with 16S rRNA methylase genes were ESBL producers, blaCTX-M-14-like, blaCTX-M-like and blaSHV-12-like were predominant genotypes of ESBLs. The plasmids of 13 isolates were transferred into the recipients E. co/iJ53. PCR and sequence analysis revealed that blaCTX-M-14-like,blaCTX-M-15-like and blaSHV-12-like were co-transferred with the armA or the rmtB to the recipients. All transconjugants harbored intll and blaTEN-1. Of the 21 isolates, 14 patterns were obtained by PFGE. Conclusion Both horizontal gene transfer and clonal spread were responsible for the dissemination of the rmtB or the armA gene in Klebsiella pneumoniae.

OBJECTIVETo explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.

METHODSThe mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.

RESULTSThe expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.

CONCLUSIONSThe recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Colonic Neoplasms ; genetics ; pathology ; therapy ; Diphtheria Toxin ; biosynthesis ; genetics ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; Genomic Imprinting ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor II ; genetics ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo study the sterilizing effect and mechanism of electrolyzed-oxidizing water (EOW) and electrolyzed-reductive water (ERW) for Bacillus subtilis var. niger (ATCC9372) and Escherichia coli (8099).

METHODSThe generations of EOW and ERW were made in the ion membrane electrolysis cell. The sterilization manner was the suspension quantitative germicidal test.

RESULTSThe killing rate of EOW for Bacillus subtilis var. niger was 99.59% in 30 minutes and the killing logarithm value was 2.38 log cfu/ml; the killing rate of ERW for Bacillus subtilis var. niger was 94.62% in 60 minutes and the killing logarithm value was 1.27 log cfu/ml; the killing rate of ERW for Escherichia coli was 100% in 30 minutes and the killing logarithm value was 8.26 log cfu/ml. When the available chlorine content (ACC) value in EOW was 74.90 mg/L and killing time was 30 minutes, the killing rate for Bacillus subtilis var. niger was 99.89% and the killing logarithm value was 2.67 log cfu/ml. When the ACC value was 6.82 mg/L, the killing rate for Bacillus subtilis var. niger was 83.30% and the killing logarithm value was 0. 78 log cfu/ml under the same time. When the oxidizing-reductive potential (ORP) and pH values of EOW were 1138 mV and 2.24 respectively, the killing rate for Bacillus subtilis var. niger was 99.99%. When the ORP and pH values of EOW were 883 mV and 5. 43 respectively, the killing rate of Bacillus subtilis var. niger was 99.73%. When the ORP value of ERW is -918 mV, the sterilizing rate for Bacillus subtilis var. niger was 94.62%; when the ORP value is -155 mV, the sterilizing ratio was only 40.19%.

CONCLUSIONIt indicates that the sterilizing mechanism of EOW is mainly chemical processes (ACC), while the physical factors are auxiliary. The sterilizing mechanism of ERW is physics sterilizing that the mainly factor is ORP.

Electrolysis ; Sterilization ; methods ; Water ; chemistry

OBJECTIVETo evaluate the detection and accuracy of fetal echocardiography for congenital heart defects among high-risk populations.

METHODSA prospective observational study of prenatal diagnosis of congenital heart disease was conducted in two tertiary obstetrics and gynecology hospitals between January 2003 and December 2004. Consecutive fetuses at risk of congenital heart disease underwent detailed fetal echocardiography during the study period. B-mode and colour/pulsed Doppler flow imaging were used in all cases. Follow-up was sought for all pregnancies. Indications for referral, maternal and gestational age at diagnosis, as well as prenatal and postnatal diagnosis were recorded prospectively. By comparing prenatal and postnatal diagnoses, sensitivity, specificity, and predictive values were estimated.

RESULTSA series of 2063 high-risk fetuses underwent detailed fetal echocardiography during the study period. The mean gestational age at examination was 26.5 weeks, ranging from 16 to 42 weeks. The most common indications for fetal echocardiography were advanced maternal age (31.7%), fetal arrhythmias (13.5%) and maternal infections (10.4%). Forty-three cases of fetal congenital heart disease were detected. The mean gestational age at prenatal diagnosis was 27.3 weeks ranging from 16 to 40 weeks. There were 3 false-negatives and 1 false-positive. The sensitivity, specificity, positive and negative predictive values were 92.1%, 99.9%, 97.2%, and 99.8%, respectively. Diagnostic accuracy was 86.1%. A cardiac defect suspected on routine prenatal sonography accounted for the highest proportion of abnormal cases (67.4%). As for pregnancy outcome, there were 24 (52.1%) terminations; 2.2% died in utero, 13% postnatally, and 28.3% survived.

CONCLUSIONS(1) Fetal congenital heart disease can be identified reliably by prenatal echocardiography. (2) Possible congenital heart disease or suspected heart defect noted on a screening obstetric sonogram is an important indication for fetal echocardiography. (3) A sequential segmental approach is critical for correct evaluation of the cardiac malformation. (4) The outcome of the patients with congenital heart disease is poor and a multidisciplinary approach is needed to the parental counseling and perinatal management planning.

China ; epidemiology ; Early Diagnosis ; Female ; Fetal Heart ; abnormalities ; diagnostic imaging ; pathology ; Fetus ; abnormalities ; Follow-Up Studies ; Gestational Age ; Heart Defects, Congenital ; diagnostic imaging ; epidemiology ; pathology ; Humans ; Mass Screening ; Pregnancy ; Prospective Studies ; Sensitivity and Specificity ; Ultrasonography, Prenatal

OBJECTIVETo evaluate the prenatal diagnostic accuracy of fetal echocardiography for congenital heart defects.

METHODSFetal echocardiographic databases from 2001 to 2007 were searched for patients with a prenatal diagnosis of congenital heart defect, medical records were obtained and the prenatal echocardiographic findings were correlated with postnatal echocardiography results or autopsy findings, if the pregnancy was terminated or the fetus died in utero.

RESULTSPrenatal diagnosis of congenital heart defects was made in 113 pregnancies at a mean gestational age of 26.8 weeks. Pathology or postnatal echocardiography was available in 79 cases (70%) and the accuracy of prenatal diagnosis was 86% (68/79). Prenatal diagnosis was accurate in 24 of 31 patients (77%) with conotruncal malformations, 26 of 27 patients (96%) with septal defects, 9 of 10 patients (90%) with valve abnormalities, and 5 of 6 patients (83%) with univentricular hearts. There were 4 false-positives and the positive predictive value was 95% (75/79).

CONCLUSIONFetal echocardiography is a reliable tool for prenatal diagnosis of congenital heart defects despite limitations for correctly diagnosing some specific fetal heart defects.

Echocardiography ; Female ; Fetal Heart ; diagnostic imaging ; Heart Defects, Congenital ; diagnostic imaging ; Humans ; Pregnancy ; Pregnancy Trimester, Second ; Pregnancy Trimester, Third ; Retrospective Studies ; Ultrasonography, Prenatal