AIM: To investigate the expression of vascular endothelial growth factor receptor-3 (VEGFR-3) and CD31 in relation to metastasis in ovarian epithelial tumors. METHODS: VEGFR-3 and CD31 expression were examined by immunohistochemical methods, VEGFR-3 positive microlymphatic count (MLC) and microvessel density (MVD) were assessed by the image analysis. RESULTS: Both MLC and MVD in ovarian epithelial carcinomas were higher than those in benign tumors(MLC, P 0 05). CONCLUSIONS: VEGFR-3 and CD31 expression correlate significantly with metastasis, MLC and MVD might predict peritumoral lymphangiogenesis and angiogenesis, which may be as a biologic marker for metastasis in ovarian epithelial tumors. [

Objective To investigate the effect of Helicobacter pylori ( Hp) infection on the growth and nutritional status in children.Methods A total of 174 children with Hp infections were collected from Jinhua Hospital of Zhejiang University during March 2010 to September 2012, and 100 healthy children were also enrolled as the controls.The differences in age and gender between Hp-infected group and control group were not significant.All Hp-infected children were given first-line anti-Hp therapy and followed-up for two years.t test, repeated measure ANOVA and LSD test were used to analyze the growth and nutritional status between Hp infected children and healthy controls, as well as between HP-infection eradication group and relapse group.Results Among 174 Hp-infected children, 2 were diagnosed as true precocious puberty, 6 abandoned treatment and 8 were lost to follow-up.Among 158 children who completed the study, Hp infection was eradicated in 128 (eradication group), and relapsed in 30 (relapse group).The height, weight, peripheral levels of hematoglobin ( Hb) , Albumin ( Alb) , blood urea nitrogen ( BUN) , Fe and Zn in 158 Hp-infected children at the baseline were significantly lower than those in the healthy control group (t=2.674, 1.657, 12.709, 3.662, 4.227, 4.210 and 14.820, all P <0.05).The height, weight, peripheral levels of Hb, Alb, BUN, Fe and Zn in eradication group were increased in 1-and 2-year of the follow-up (F=8.350, 14.998, 50.875, 37.584, 22.701, 8.295 and 41.791, all P<0.01), while there were no significant increase in the levels of Hb, Alb, BUN and Fe in the relapse group (F=1.826, 1.659, 2.613 and 2.495, all P>0.05).In the second year of the follow-up, the increases of Alb, BUN, Fe and Zn in eradication group were significantly higher than those in the relapse group ( t=7.86, 5.17, 8.80, 5.92, 2.17 and 7.28, all P <0.05).Conclusion Hp infection may affect the growth and nutritional status of children, and the eradication of Hp infection may help to improve the development and nutritional status of the children.

Objective To study the transfeetion efficiency and safety of liposome microbubble(LM)on red fluorescent protein(RFP)in vitro and in vivo under ultrasound mediated gene transfection(USMGT)conditions.Methods Plasmids containing RFP were added to cultured Hela cells followed by ultrasound (US)exposure with LM.Different concentration of LM,US intensity and exposure time were optimized.Transfection efficiency was evaluated by fluorescent microscopy and FACS.Cell viability was verified by propidium iodide assay.In transplanted tumors in vivo study,LM and plasmid(P)were injected into the nude mice followed by US exposure(P+LM+US group).Nude mice undergoing plasmid injection alone(P group),plasmid injection and US exposure(P+US group)and plasmid and LM injection(P+LM group)were used as controls.Frozen section and histological examination were conducted and RFP expression was evaluated.Results LM and US exposure significantly increased transfeetion efficiency in cultured Hela cells (P＜ 0.01).Transfection efficiency was the most prominent under the condition of US intensity of 1.0 W/cm2 with 6%LM,duration 3 min.No apparent cell damage was found in the all groups.In transplanted tumors,strong RFP was seen in P+LM+US group.It was significantly higher than in any other groups(P＜0.0 1).No tissue damage was seen histologically.Conclusions LM could enhance USMGT effectively without causing any apparently adverse effect in vitro and in vivo.This method would be a novel,effective,safe non-viral gene transfection method and provide an alternative to current clinical gene therapy.

Objective To explore the changes and significance of serum tumor necrosis factor-?(TNF-?),interleukin-1?(IL-1?),IL-6 and IL-10 in rats with acute necrotizing pancreatitis(ANP).Methods Sixty-four Wistar rats were randomly divided into 2 groups: sham operation group(SO group n=32) and ANP group(n=32).The ANP model was established by using retrograde injection of Sodium Taurocholate into cholangiopancreatic duct.The changes of serum endotoxin(ET),TNF-?,IL-1?,IL-6 and IL-10 in different groups and different time points were observed.Results The levels of serum ET,TNF-?,IL-1?,IL-6 and IL-10(except IL10 of ANP group in 16h) in rats of ANP group were significantly higher than those of SO group(P

Objective To probe into the clinical value of left atrial(LA) volme tracking method (LAVT) on the assessment of left atrial function in patients with hypertension. Methods Thirty-one patients with hypertension and 31 control subjects were involved. Images of apical two-chamber and four-chamber views were obtained by two-dimensional echocardiography. LA maximal volume (LAVmax), LA minimal volume (LAVmin), LA presystolic volume (LAVpre), systolic LA filling rate (dv/dtS), early diastolic LA emptying rate (dv/dtE) and late diastolic LA emptying rate (dv/dtA) were derived using LAVT. LA passive emptying volume(LAVp), LA passive emptying fraction(LAVpEF), LA active emptying volume(LAVa), LA active emptying fraction (LAVaEF), LA total emptying volume (LAVt), LA total emptying fraction(LAVtEF) were calculated. All volume measurements were corrected to body surface area (LAVI). Results There were significant differences in the LAVImax, LAVImin, LAVIpre, LAVpEF, LAVIa, LAVIt,dv/dtS, dv/dtA between hypertensive patients and control subjects(P <0.05~0.001), but no significant differences were found in the LAVp, LAVaEF, LAVtEF,dv/dtE between the two groups. The LAVImax has shown significant positive correlation with LAVIp, LAVIa and LAVIt(r = 0.588~0.812, P <0.001). LAVaEF has shown positive correlation with the LAVtEF(r = 0.833, P<0.001), whereas LAVpEF has not(r = 0.420, P <0.01). There was a significant positive correlation with LAVImax and LAVIpre(r = 0.908, P <0.001), LAVIpre and LAVIa(r = 0.689, P <0.001) ,dv/dtE and LAVIp(r = 0.690, P <0.001),dv/dtA and LAVIa(r = 0.600, P <0.001). Conclusions In hypertensive patients,left atrial reservoir function and booster pump function increase, while left atrial conduit function decreases. LAVT has a potential ability to evaluate left atrial function.

To investigate the effects of fluoride on the expression of thyroglobulin (TG),thyroid peroxidase(TPO),odium iodide symporter (NIS) genes in FRTL cells,FRTL cells cultured in vitro were treated in logarithmic phase with sodium fluoride at different concentration of 20.0,10.0,5.0,2.5,1.25 mg/L,respectively.After 72 h,the cells were collected and semi-quantitative RT-PCR for TG mRNA,TPO mRNA and NIS mRNA and β-actin was performed.The results showed that compared with the control cells,the expression levels of TG gene in FRTL cells treated with lower concentration of sodium fluoride increased compensatively,but decreased significantly (P<0.05) with higher concentration(>5.0 mg/L);the RT-PCR products of TPO and NIS in FRTL cells treated with all concentration of sodium fluoride reduced,some of them were significant (P<0.05).It is concluded that fluoride can reduce the expression of TG,TPO,NIS genes in thyroid cells,conseguently cause dysfunction" of thyroid in uptaking and unilizing of iodine,and synthesizing,storing,secreting of thyroid hormones.

Objective To investigate the value of time parameters(the transit time in coronary and the half filling time in myocardium)in assessing the blood flow of ischemia myocardium through establishing canine model of acute myocardial infarction.Methods Eighteen healthy anesthetized open-chest dogs were ligated the left anterior descending(LAD),3 hours later the contrast agent(C3F8)was injected into femoral vein in a fixed velocity to perform myocardial contrast echocardiography(MCE)examination.The first perfusion and the replenishment processes were recorded by real-time tri-plane MCE(RT-TP-MCE)and real-time single-plane MCE(RT-SP-MCE)respectively.both the two different processes were analyzed by the function Y=A ×(1-e-βt)+B.RT-SP-MCE used A(plat of peak intensity)and β(slope rate of the curve)values to quantitatively calculate the myocardial blood flow(MBF=A×β).The time point of the first microbubbles coming into the left ventricle was normalized to O in RT-TP-MCE,the time point of the first microbubbles coming into the myocardium was defined as the transit time in coronary(Tc),and the time point when microbubbles in myocardium achieved the 50% plat video intensity was defined as the half filling time in myocardium(Tm).Evans blue dye and TTC staining were performed to identify normal.ischemic and infarct myocardium.Results The time parameters of the two groups-normal myocardium and ischemic myocardium from RT-TP-MCE were:Tc(10.1±1.3)s and(20.4±7.1)s(P＜0.01),Tm(17.1±2.2)s and(39.7±8.8)s(P＜0.01).There were significantly negative correlation between the time parameters(Tc,Tm)and MBF from RT-SP-MCE in ischemic myocardium(r=-0.876,P＜0.01;r2=-0.894,P＜0.01).Conclusions The first perfusion imaging with RT-TP-MCE could be used to simultaneously,quantitatively evaluate the myocardial perfusion.The lower flow,the longer transit time in coronary and half filling time in ischemic myocardium.

Objective To investigate the elasticity of the radial artery wall in type 2 diabetes mellitus (T2DM) patients with elastosonography. Methods A total of 37 patients with T2DM and 42 normal subjects were studied with elastosonography. The systolic diameter (Ds) and diastolic diameter (Dd) of the radial artery were measured, and the strain ratio of the blood in the radial artery to the wall of the radial artery was calculated. Results The strain ratio of the radial artery in T2DM group was significantly higher than that in normal group (P＜0.05). There was no significant difference in Ds and Dd of the radial artery between T2DM group and the control group (P＞0.05). Conclusion The early change of the radial artery wall elasticity in patients with T2DM can be assessed with elastosonography.

Objective To evaluate the collateral circulation after canine acute coronary artery occlusion with contrast real-time three-dimensional echocardiography (RT-3DE). Methods Left anterior descending coronary artery was ligated in 12 healthy mongrel dogs. All dogs underwent myocardial contrast echocardiography (MCE) before ligation, immediately after ligation, at 30 min and 180 min after ligation respectively, and myocardial mass with collateral supply was calculated. Results Different degrees of collateral circulation were established in all dogs after left anterior descending coronary artery ligation. At 30 min after ligation, the myocardial mass with collateral supply was (9.65±2.90) g, while at 180 min after ligation were (12.58±3.98) g (P<0.01). Conclusion The coronary collateral circulation can be observed clearly and myocardial mass with collateral supply can be quantified accurately with contrast RT-3DE.

In this study, we evaluated the development potential of caprine mammary gland epithelial cells (CMGECs) after transfection and nuclear transfer into enucleated, ovulated oocytes. We first isolated CMGECs from udders of lactating goats which were transfected with expression plasmid for human lacterrin and selected by G418. Then we chose sixteen neomycin resistant lines and induced them with prolactin for the expression of human lactoferrin checked by Western blotting. The donor cells, expressing human lactoferrin of 75 kD, were fused and activated with enucleated ovulated oocytes. Pronuclear-stage reconstructed embryos were transferred into the oviducts of 16 recipient goats. There were fourteen (87.5%), thirteen (81.3%), and ten (62.5%) pregnancies confirmed pregnant by ultrasound on Day 30, 60, and 90, respectively. Three recipients carried the pregnancies to term and delivered one goat each. Nested PCR-RFLP analysis confirmed that all of the kids were clones of the donor cells. These results demonstrated that CMGECs after transfection remain totipotent for nuclear transfer.
Animals ; Cloning, Organism ; methods ; Epithelial Cells ; cytology ; Female ; Goats ; Humans ; Lactoferrin ; biosynthesis ; Mammary Glands, Animal ; cytology ; Nuclear Transfer Techniques ; Pregnancy ; Transfection