Electrocardiography ; Female ; Fetal Hypoxia ; complications ; Humans ; Infant, Newborn ; Myocardial Infarction ; etiology ; therapy ; Myocardium ; metabolism ; pathology ; Treatment Outcome

Background Studies showed that microRNA (miR)-375 suppresses the growth,apoptosis,migration and adhesion of tumor cells,and it plays a regulation to the changes of vascular endothelial growth factor (VEGF) in tumor tissue to arrest neovascularization.However,whether miR-375 intervenes the formation of new blood vessel in eyes is unelucidated.Objective This study was to explore the effects of miR-375 on human retinal capillary endothelial cells (HRCECs) function induced by hypoxia.Methods HRCECs were cultured using IMDM and divided into normal control group,CoCl2 model group,CoCl2 +miR-375 mimic group,CoCl2 +miR-375 mimic control group,CoCl2+miR-375 inhibitor group and CoCl2 +miR-375 inhibitor control group,and hypoxia cell models were created by adding 200 μmol/L CoCl2.MiR-375 and frizzled 4 (FZD4) small interfering RNA (siRNA) were transfected into the cells by 50 nmol/L miRNA lipidosome for 48 hours.The proliferation of the cells was detected by MTT assay;migrated number of the cells was examined by Transwell chamber assay;ELISA was employed to detect the concentrations of VEGF and VE-cadherin in the medium;the expression of β-catenin,cyclinD1,matrix metalloproteinase-2 (MMP2) and VEGF proteins were analyzed by Western blot;tube length of vessel formation was evaluated by Matrigel assay.Cultured cells were divided into normal control group,CoCl2 model group,CoCl2 +mock group and CoCl2 + FZD4 siRNA group,the relative expression of FZD4,a miR-375 targeted gene,was detected by luciferase reporter.Results The relative expression of miR-375 mRNA was significantly increased in the CoCl2 +miR-375 mimic group compared with the CoCl2 + miR-375 mimic control group and reduced in the CoCo2 + miR-375 inhibitor group compared with the CoCo2 + miR-375 inhibitor control group (t =-19.237,8.764,both at P＜0.01),with a higher transfected efficacy for miR-375.The cell proliferative fold,migrated cell number,VEGF and VE-Cadherin contents in the medium and the tube length were significantly different among the CoCl2 model group,CoCl2 +miR-375 mimic group,CoCl2 +miR-375 mimic control group,CoCl2 +miR-375 inhibitor group and CoCl2 +miR-375 inhibitor control group (F =24.324,26.776,14.113,19.225,15.040,all at P＜0.001),and those in the CoCl2 +miR-375 mimic group were evidently reduced in the CoCl2 +miR-375 mimic group compared with the CoCl2 +miR-375 mimic control group,while those in the CoCl2 +miR-375 inhibitor group were considerably elevated in comparison with the CoCl2 +miR-375 inhibitor control group (all at P＜0.01).The expressions of β3-catenin,cyclinD1,MMP2 and VEGF protein were significantly different among the normal control group,CoCl2 model group,CoCl2 +miR-375 mimic group,CoCl2+miR-375 mimic control group,CoCl2 +miR-375 inhibitor group and CoCl2 +miR-375 inhibitor control group (F=11.753,13.283,16.770,10.334,all at P＜0.001).In addition,the cell proliferative fold,migrated cell number and the tube length were significantly increased in the CoCl2 model group and CoCl2+mock group,and those in the CoCl2+FZD4 siRNA group were decreased in comparison with the CoCl2 +mock group (all at P＜0.05).Conclusions MiR-375 inhibits the growth,migration and tube formation ability of HRCECs in hypoxic status probably by regulating the activation of Wnt pathway via directly targeting FZD4.

Objective To explore the effect and potential mechanism of suppressors of cytokine signaling 3 (SOCS3) knockout on cell viability and apoptosis of retinal ganglion cells (RGCs) after optic nerve injury.Methods The optic nerve transection was used to construct optic nerve injury model of rats,and RGCs were isolated after optic nerve injury.The experimental animals were divided into optic nerve transection injury (ONT) group and sham-operation (Sham) group.The expression of SOCS3 in RGCs was detected by Western blot and RT-PCR in each group.Subsequently,SOCS3 siRNA was transfected into RGCs of Sham group and ONT group,and the experimental were further subdivided into blank control group,negative control group and SOCS3 silence groups.Cell viability was measured by CCK8 and MTr methods.Apoptosis was detected by Hoechst 33342 staining and flow cytometry.Furthermore,the mTOR siRNA and SOCS3 siRNA were co-transfected into RGCs,and cell viability and apoptosis were detected.Results The expression of SOCS3 was dramatically increased at 3 days after injury in the ONT group when compared with Sham group (P =0.049),and it showed an increased tendency gradually along with the extension of injury time.Compared with the blank control in the ONT group,SOCS3 silence markedly promoted cell viability [(49.47 ± 7.35) ％ vs.(73.24 ± 8.70) ％],reduced cell growth inhibition [(27.25 ±0.75)％ vs.(10.96 ± 1.07)％] and apoptosis [(23.06 ± 1.43)％ vs.(10.65 ± 1.77)％].The result of Hoechst 33342 staining indicated that SOCS3 silence ameliorated the cell apoptosis induced by ONT.In addition,SOCS3 silence significantly improved pS6 expression at 2 weeks after injury,and mTOR and SOCS3 co-silence reduced cell viability,increased cell growth inhibition and apoptosis compared with SOCS3 silence group after injury.Conclusion SOCS3 silence promotes injury-induced cell viability of RGCs and suppresses injury-induced apoptosis of RGCs via up-regulating mTOR activity in the later period of injury.

Objective To study on ERCP findings correlating with tumor size and staging of pancre- atic cancer(PC).Methods We selected 35 patients with PC who were identified by surgery and/or pathol- ogy and had undergone ERCP with clear imaging findings.The lengths of pancreatic duct and common bile duct stricture as well as the diameter of upstream dilated pancreatic or bile duct were measured on ERCP films.The ralationship among ERCP findings and tumor size and JPS T-stage of PC were analysed by correla- tion analysis.Results Of the 35 patients with PC,ERCP showed the pancreatic duct stricture in 25 and cut- off of pancreatic duct in 10(7 of them,the lesion lengths were revealed after a guide wire advanced beyond the obstructive site)and common bile duct stricture in 31.The correlation coefficients between tumor size and lengths of pancreatic duct or common bile duct stricture were 0.709(P=0.0001)and 0.540(P=0.0053) respectively.The correlation coefficient between JPS T-stage and lengths of pancreatic duct stricture was 0.514(P=0.0052).Conclusion The length of pancreatic duct stricture in ERCP was positively correlated with the tumor size and staging of PC.But it is not consistent when the length of stricture is rather short.The lengths of bile duct stricture also have some effects on predicting the tumor size of pancreatic head cancer.

Objective To investigate the feasibility of three-dimensional contrast-enhanced ultrasonic cholangiography(3D-CEUSC) in isolated porcine liver specimens.Methods The lobes of 5 isolated porcine livers were taken as scanning units and the lobular bile branches were cannulated.3D-CEUSC was performed after the administration of diluted ultrasonic contrast agents(SonoVue) through the cannula.The images were assessed in term of maximum visualization of branching orders and image quality.The coincidence of morphous was compared between 3D-CEUSC and cholangiagraphy using fluoroscopy.Results Fourteen lobular biliary branches were cannulated successfully in 5 isolated porcine livers.The maximum visualization of branching orders from the third- to first-order branches were 42.9% (6/14) ,42.9% (6/14), 14.2% (2/14) in sequence.The image quality was 21.4% (3/14) graded as well, 57.2% (8/14) as moderate, and 21.4% (3/14) as poor.Compare with cholangiagraphy using fluoroscopy, the coincidence of morphous was 42.9%(6/14) for well,42.9% (6/14) for moderate,and 14.2% (2/14) for poor.Conclusions 3D-CEUSC could be a new imaging method to evaluate the biliary anatomy.

Objective To assess the value of three-dimensional contrast-enhanced ultrasound (3D-CEUS) in evaluation origin of segment Ⅳ feeding artery in living liver donors. Methods 3D-CEUS were used to examine 20 living liver donors. The origin of segment Ⅳ feeding artery were evaluated by two radiologists who blinded to other imaging information. The findings obtained on operation and (or) CTA were used as the reference standard. Results Segment Ⅳ feeding artery originating from left artery, both left and right artery, proper hepatic artery were demonstrated on 3D-CEUS in 13, 1 and 2 patients,respectively. 3D-CEUS accurately depicted the origin of segment Ⅳ feeding artery in 16(80%) patiens.Conclusions 3D-CEUS could be a useful method to demonstrate the origin of segment Ⅳ feeding artery in preoperative evaluation for LDLT.

efore graft harvesting in living donor liver transplantation.

Objective To investigate the feasibility and methodology of detecting the microcirculation of hilar bile duct wall with contrast-enhanced ultrasound (CEUS). Methods Sixteen patients after orthotropic liver transplantation(OLT) were studied. The dosage of contrast agent(SonoVue) was set for 2 groups: 1.5 ml and 2.4 ml. Each group was composed of 8 patients. The imaging quality of CEUS were retrospectively analyzed and divided into 3 grades: good, ordinary and bad. There were 2 patients received retransplantation. Their morphologic features of hilar bile duct wall in baseline ultrasound (US) and CEUS were compared with those in pathological examination. Results Biliary perfusion could be displayed by CEUS continuously,real-timely and clearly. The imaging quality was good in 8 cases(50%), ordinary in 6 cases (37. 5%) and bad in 2 cases(12. 5%). There was no significant difference between the imaging quality of 1. 5 ml and that of 2. 4 ml group ( P = 0. 78). Conclusions The imaging quality of hilar bile duct wall in CEUS is good enough to detect its microcirculation. There is no significant difference between the imaging quality of 1. 5 ml and that of 2.4 ml group.

Objective To investigate the role of intra-operative and percutaneous three-dimensional contrast-enhanced ultrasonic cholangiography (3D-CEUSC) in the diagnosis of biliary anomalies. Methods Diluted SonoVue, an ultrasonic contrast agent, was injected into the cystic duct of 13 living liver donors via a cannula for intra-operative 3D-CEUSC, while injection via a T tube or a PTCD tube were done in 14 patients for percutaneous 3D-CEUSC. The maximum branching order of the intra-hepatic bile ducts, the percentages of display of the first-, second-, and third-order intra-hepatic bile ducts, and the rates of detection of biliary anatomical anomalies were evaluated. Results The median of the maximum branching order of the intra-hepatic bile ducts shown was fourth-order on intraoperative 3D-CEUSC and third-order on percutaneous 3D-CEUSC, respectively. The difference was significant (P=0.01). From the first- to the third-order of intra-hepatic bile ducts, the percentages of display for intra-operative 3D-CEUSC were 100% (23/23), 96.2% (50/52) and 82.7% (86/104),respectively, while they were 100% (24/24), 94.6% (54/56) and 60. 7% (68/112) for percutaneous 3D-CEUSC. The differences were not significant when comparing the first- and the second-order branches of intra-hepatic bile ducts as shown either on intra-operative or percutaneous 3D-CEUSC (P=1).The percentages of display of the third-order branches using intra-operative 3D-CEUSC was higher than that of percutaneous 3D-CEUSC (P＜0. 0001). Three biliary anatomical anomalies were detected by intra-operative 3D-CEUSC and 4 anomalies were detected by percutaneous 3D-CEUSC. All of these anomalies were confirmed by X-ray cholangiography. Conclusion Both intra-operative and percutaneous 3D-CEUSC could diagnose biliary anatomical anomalies accurately. These procedures may become important techniques to evaluate the biliary anatomy in hepatobiliary surgery pre-operatively or intraoperatively.

Objective To assess the value of three-dimensional contrast-enhanced ultrasonography (3D-CEUS) in evaluation of hepatic arteries variants.Methods Both two-dimensional contrast-enhanced ultrasonography (2D-CEUS) and 3D-CEUS were used to assess 30 patients including living donor candidates and patients with upper abdomen tumors.The reference standard was operation or CTA or DSA or MRA,and the accuracy for detecting hepatic artery variants provided by the two methods was evaluated.Arterial anatomic types were defined by using Michels classification.Results The total accuracy for detecting hepatic artery anatomy types by 2D-CEUS was 40.0% (12/30),while 83.3% (25/30) by 2D-CEUS.For convention anatomy types the accuracy on 2D-CEUS and 3D-CEUS were 40.9%(9/22)and 90.9%(20/22),respectively.The difference was statistically significant (P<0.05).For anatomy variants types the accuracy on 2D-CEUS and 3D-CEUS were 37.5%(3/8)and 62.5%(5/8),respectively.No significant difference between these two methods was observed.Conclusion 3D-CEUS was a new method in diagnosis of hepatic arteries anatomy types with practical clinical value in evaluation of the living liver donors.