Objective: To screen for miRNAs potentially involved in the pathogenesis of renal ischemia/reperfusion injury. Methods: Renal ischemia reperfusion injury model was established with SD rats. Twelve hours after reperfusion, blood urea nitrogen and serum creatinine were determined and miRNA expression in the kidney was detected using miRNA microarray. Bioinformatics methods were used for a preliminary analysis of potential targets for differentially expressed miRNAs. Results: Blood urea nitrogen and serum creatinine were both elevated 12 h after reperfusion. miRNA microassay showed 36 aberrantly expressed miRNAs, with 15 miRNAs having an expression level higher than 2 folds. Results of real-time PCR were generally in accordance with the microarray results. The elevated miRNAs included miR-290, miR-894, miR-292-5p, miR-327, miR-374, miR-98, miR-352, miR-132, miR-146b and miR-196a; and the down-regulated miRNAs included miR-145, miR-329, miR-375, miR-140 * and miR-29a. Bioinformatics showed that these miRNAs were related to inflammation, cell death and proliferation, angiogensis and fibrosis. Conclusion: Several miRNAs are aberrantly expressed during renal ischemia/ reperfusion injury, which may influence renal injury through regulating inflammation, cell death and proliferation, angiogensis and fibrosis, but the exact mechanism remains to be further investigated.

OBJECTIVETo study value of velocity vector imaging (VVI) in assessment of normal left ventricular diastolic function and the corresponding reference values.

METHODSNinety-seven healthy subjects were selected by combined clinical, ultrasound, and NT-proBNP examinations. Using a Siemens Acuson Sequoia C512 echocardiograph, VVI was adopted to examine the myocardial early diastolic velocity (E) of the septal, lateral, anterior, inferior, anterior septum, and posterior wall of the left ventricle at the level of mitral annulus. The images were analyzed for VVI and the mean diastolic velocity (E(m)) and hence the E/E(m) ratio was calculated based on the offline workstation interface.

RESULTSThe reference range of E/E(m) ratio derived from the data of the 97 healthy subjects was (unilateral boundaries with 95% limit) and (1, 22.935), was (1, 22.300) in male subjects and (1, 24.766) in female subjects. The reference E/E(m) range was (0, 22.413) in male subjects under 50 years of age, (1, 24.766) in female subjects under 50 years, (1, 22.300) in male subjects over 50 years, and (1, 24.766) in female subjects over 50 years.

CONCLUSIONVVI is a good method for non-invasive evaluation of the left ventricular diastolic function and provides an accurate and reliable means for clinical assessment of the left ventricular diastolic function.

Adolescent ; Adult ; Aged ; Blood Flow Velocity ; physiology ; Diastole ; physiology ; Echocardiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Reference Values ; Ventricular Function, Left ; physiology ; Young Adult

OBJECTIVETo investigate the effect of therapeutic ultrasound-induced microbubble destruction on the microcirculation of rat skeletal muscle.

METHODSThirty SD rats were randomized into 5 groups (n=6), namely normal saline, microbubble, ultrasound, high-energy ultrasound microbubble and low-energy ultrasound microbubble groups. Before and after the treatments, the diameter and blood flow velocity in the microvessels in the skeletal muscle were measured, and the structural changes of the injured microvessels observed by electron microscopy.

RESULTSMicrobubble cavitation did not produce significant effect on the mean arterial pressure and diameter of microvessels in rat skeletal muscle (P>0.05), but the blood flow velocity was obviously lowered and blood flow volume reduced in the microvessels. The reduction of the flow velocity and blood flow volume and their subsequent recovery were associated with ultrasound energy, and in the low ultrasound energy group, the flow velocity and blood flow volume in the of venules recovered obviously after about 15 min, which, however, took approximately 1 h for the arterioles. In contrast, recovery of the flow velocity and blood flow volume in the microvessels took more than 2 h in the high ultrasound energy group. Cavitation resulted in endothelium cell rupture, widening of the endothelial interspace and entry of the red blood cells into the extravascular tissues as revealed by electron microscopy, but no rupture of the lining endothelium was observed 2 h after the treatment.

CONCLUSIONSEndothelium cell rupture induced by microbubble cavitation may affect the local microcirculation, and lower ultrasound energy exposure is associated with milder endothelial injury and more rapid recovery.

Animals ; Blood Flow Velocity ; Blood Vessels ; pathology ; physiopathology ; Endothelial Cells ; pathology ; ultrastructure ; Female ; Male ; Microbubbles ; Microcirculation ; Microscopy, Electron ; Microspheres ; Muscle, Skeletal ; blood supply ; Rats ; Rats, Sprague-Dawley ; Ultrasonics

OBJECTIVETo assess the value of velocity vector imaging (VVI) and quantitative tissue velocity imaging (QTVI) in assessing left ventricular diastolic function of the dogs with acute myocardial ischemia.

METHODSSix healthy mongrel dogs were subjected to ligation of the left circumflex artery or left anterior descending artery to induce coronary artery stenosis of varying degrees. The mean peak diastolic velocity (Em) of the ventricular walls around the mitral annulus was recorded with VVI or QTVI in the coronary blood flow. The left ventricular end diastolic pressure (LVEDP) was measured with pigtail catheter in the left ventricle.

RESULTSAs the coronary blood flow decreased, LVEDP was gradually increased, and Em measured by VVI or QTVI were also gradually decreased. A good linear correlation was shown between Em measured by VVI or QTVI and LVEDP (r=-0.834, P<0.001, and r=-0.68, P<0.001, respectively). A significant difference was observed in the correlation coefficient between VVI and QTVI (Z=2.625, P=0.0087).

CONCLUSIONVVI and QTVI both provide good noninvasive means for measuring left ventricular diastolic function. VVI, a new echocardiographic modality without angular dependence, is better than QTVI in evaluating left ventricular diastolic function.

Animals ; Disease Models, Animal ; Dogs ; Echocardiography ; methods ; Male ; Myocardial Ischemia ; diagnostic imaging ; physiopathology ; Ventricular Function, Left

OBJECTIVETo investigate the impact of high-dose microbubbles induced by high mechanical index myocardial contrast echocardiography (MCE) on vascular permeability and its recovery time in rats.

METHODSThirty male Wistar rats were randomized into 4 MCE groups (groups A-D) and a control group. In the MCE groups, Evans blue was injected at 10 s before MCE (A), immediately after the end of MCE (B), and at 5 min (C) and 20 min after the end of MCE (D). In the control group, the microbubbles and Evans blue were injected at the end of a 5-min ultrasound exposure. All the rats were sacrificed 5 min after Evans blue injection, and the content of Evans blue in the myocardium and the percentage of Evans blue leakage area were determined.

RESULTSThe percentage of Evans blue leakage area in groups A, B and C were significantly higher than that in the control group (P<0.05), while the percentage was similar between group D and the control group (P>0.05). Evans blue contents in groups A and B were significantly higher than that in the control group (P<0.05), but groups C and D showed comparable contents with the control group E (P>0.05). No significant changes of the heart rates and premature beat number were observed during and after MCE in these groups (P>0.05).

CONCLUSIONHigh mechanical index MCE and a high contrast dose may induce increased microvascular leakage in rats, and the vascular permeability can recover in 20 min after MCE.

Animals ; Capillary Permeability ; drug effects ; Contrast Media ; pharmacology ; Coronary Vessels ; physiopathology ; Echocardiography ; Male ; Microbubbles ; Rats ; Rats, Wistar

OBJECTIVETo investigate the drag-reducing effect of polyethylene oxide (PEO) on the velocity of red blood cells in rat cremaster microcirculation.

METHODSBlood samples were collected from 6 Wistar male rats (100-110 g) via the post-orbital venous plexus. The red blood cells were separated by centrifugation and labeled by fluorescinisothiocyate (FITC). After successful establishment of cremaster model, the labeled red blood cells were injected into the jugular vein, and the microcirculation was observed and recorded under fluorescence microscope. The hemodynamic parameters and microcirculation video was recorded every 4 min since 4 min before PEO or normal saline injection. Both PEO (10 ppm) and normal saline was injected into the same rat in random sequence at a constant rate of 3.5 ml/h for 20 min followed by observation for another 20 min. The velocity of the labeled-red blood cells was determined by IPP 6.0 software.

RESULTSCompared with normal saline, PEO significantly increased the velocity of the red blood cells in the rat cremaster microcirculation (498.7-/+182.89 microm/s vs 773.54-/+308.27 microm/s, P=0.012). No significant changes in the heart rate and arterial blood pressure were observed during the experiment (P=0.836, P=0.420).

CONCLUSIONPEO at an extremely low concentration can significantly increase the velocity of the red blood cells in rat cremaster microcirculation and produces no significant impact on heart rate and arterial blood pressure.

Animals ; Blood Flow Velocity ; drug effects ; Male ; Microcirculation ; drug effects ; physiology ; Muscle, Smooth ; blood supply ; Polyethylene Glycols ; pharmacology ; Rats ; Rats, Wistar ; Testis

OBJECTIVETo observe the effect of polyethylene oxide (PEO) solution at different concentrations on abdominal aortic blood flow and vascular resistance in rats and evaluate the safety and drag-reducing effect of PEO solution.

METHODSThirty-two rats were anesthetized and randomly divided into 4 groups. An ultrasonic flow probe was deployed on the abdominal aorta (5 mm above the common iliac artery) to measure the blood flow. The carotid artery pressure, iliac artery pressure, iliac vein pressure, central venous pressure (CVP) and ECG were also monitored. Saline or different concentrations of PEO [(1x10(-6)(low), 1x10(-5)(middle) and 5x10(-5)(high) g/ml)] were injected in the 4 groups of rats through the caudal vein at a constant rate of 5 ml/h for 20 min, and the changes of the vascular resistance was observed. RESULTS After injections of 1x10(-6) and 1x10(-5) g/ml PEO, the abdominal aortic flow increased significantly (P<0.05) while the vascular resistance was reduced (P(low)=0.052, P(middle)<0.001) as compared to those in the saline control group. Following the injection with 5x10(-5) g/ml PEO, the abdominal aortic flow increased to a threshold in the initial 4 min, after which it rapidly decreased to approach the baseline levels despite continuous infusion. Blood pressure remained stable after the injections except for 5x10(-5) g/mlPEO injection, which resulted in a reduction of the blood pressure by about 10 mmHg (P=0.014). The heart rate and CVP both underwent no significant changes following the injections.

CONCLUSIONThe drag-reducing effect of PEO is closely related to its concentration, and compared with 1x10(-6) g/ml, 1x10(-5) g/ml PEO more effectively increases the blood flow and decreases the resistance. The effectiveness and safety of EPO are attenuated at a concentration higher than 5x10(-5) g/ml.

Animals ; Aorta, Abdominal ; physiology ; Blood Flow Velocity ; drug effects ; Dose-Response Relationship, Drug ; Male ; Polyethylene Glycols ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Vascular Resistance ; drug effects

OBJECTIVETo explore the duration of enterovirus (EV) nucleotides positive in feces samples of hand-foot-mouth disease (HFMD) patients after recovery.

METHODSA consecutive 6-week follow up were carried out towards 49 cases of laboratory-diagnosed HFMD patients. A total of 5 - 8 g feces sample was collected from each patient once a week. The common EV nucleotides of HFMD were detected by RT-PCR method and analyzed by Kaplan-Meier Survival Analysis Method.

RESULTSThe subtypes of the 49 HFMD patients included 16 enterovirus 71 (EV71), 15 coxsackievirus A16 (CoxA16) and 18 EV; a six-week follow up was carried out among all of them. In the first week, one EV71 patient and two EV patients were lost; in the fourth week, one CoxA16 were lost; and in the fifth week, one EV71 patient was lost. During the consecutive 6-week follow-up, the positive rates of EV nucleotides among EV71 patients were 81.3%, 60.9%, 47.4%, 33.9%, 27.1% and 18.1% separately; and the positive rates in CoxA16 group were 93.3%, 73.3%, 53.3%, 33.3%, 16.7% and 8.3% respectively. In EV group, the positive rates of EV nucleotides were 44.4% and 7.4% in the first two weeks and then turned to negative in the next 4 weeks. There was significant statistical difference in positive rates of EV nucleotides among different patients (χ(2) = 11.78, P = 0.001); however, each group of HFMD patients showed a declined trend with the extension of time.

CONCLUSIONThe duration of EV nucleotides positive in feces samples of HFMD patients lasted for a long period since their recovery; and the positive results in EV71 and CoxA16 patients might last for 6 weeks.

Child, Preschool ; Enterovirus ; genetics ; isolation & purification ; Feces ; virology ; Follow-Up Studies ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Nucleotides ; isolation & purification

ObjectiveTo detect the concentration and distribution of fluoride ions in osteoblasts exposed to fluoride in vitro culture,and to provide basic information for studying the effect of fluoride on osteoblast injury.MethodsIn vitro cultured osteoblasts were exposed to 0,5,10,20,40 mg/L fluoride for 3,10,30 d (n =6),respectively.Concentration and distribution of fluoride ions in the cytoplasm and the nucleus of these osteoblasts were determined by nuclear magnetic resonance spectroscopy.Results(①) After cultured for 3 d,fluoride ion content of the bone cytoplasm exposed to different concentrations of fluoride 0,5,10,20,40 mg/L were (0.83 ±0.65),(0.54 ± 0.23),(0.65 ± 0.77),(0.59 ± 0.87),(3.64 ± 1.21 )mg/L,respectively,and the values of exposed to 40 mg/L fluoride group was significantly higher than that of exposed to 0,5 mg/L groups (all P ＜ 0.05).(②)after cultured for 10 d,the composition of the fluoride ion in cytoplasm of exposed to fluoride 10,20,40 mg/L groups were (4.03 ± 1.23),(3.66 ± 0.98),(6.26 ± 2.10)mg/L,respectively,which were higher than that of exposed to 0,5 mg/L groups [(0.78 ± 0.75),(2.69 ± 0.89)mg/L,respectively,all P ＜ 0.05].Of fluoride 20,40 mg/L groups,the composition of the fluoride ion in nucleus were (1.63 ± 1.19),(2.17 ± 1.21 )mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.65 ± 0.46),(1.57 ± 0.33) mg/L,all P ＜ 0.05].(③)After cultured for 30 d,of the exposed to fluoride 10,20,40 mg/L groups,the composition of the fluoride ion in cytoplasm were (3.99 ± 0.84),(4.33 ± 1.67),(5.80 ± 1.38)mg/L,respectively,which were higher than that of 0,5 mg/L groups[(0.88 ± 0.44),(2.84 ± 0.43)mg/L,all P ＜ 0.05].The composition of the fluoride ion in nucleus of the fluoride 20,40 mg/Lgroups were (3.33 ± 1.46),(3.53 ± 1.22)mg/L,respectively,which were significantly higher than that of 0,5mg/L groups [(0.70 ± 0.66),(1.99 ± 0.76)mg/L,all P ＜ 0.05].ConclusionsWhen osteoblasts are exposed to fluoride environment,fluoride ions enter into the osteoblasts quickly,and quickly accumulate in the nucleus,showing a special affinity between fluoride and bone tissue.Intracellular fluoride ions increase with the increase of contact time and exposure dose.

OBJECTIVETo investigate the genetic abnormalities of fetuses with congenital heart diseases (CHD), and to provide guidance for the management of pregnancy and genetic counseling.

METHODSEighty-one fetuses with CHD detected by fetal echocardiography were analyzed by karyotyping after amniocentesis, cordocentesis or chorionic sampling. Then 22q11.2 deletion/duplication was detected by a competitive fluorescent multiplex short tandem repeat assay in 47 CHD fetuses without chromosomal abnormalities. With fluorescence in situ hybridization (FISH) using LSI dual color DNA probe, the deletion/duplication status was confirmed.

RESULTSThirty-four of 81 CHD fetuses had chromosomal anomalies, and 1 of the 47 CHD fetuses without chromosomal anomalies had duplication at chromosome 22q11. The incidence of aneuploidy associated CHD was 43.2%. The rate of chromosomal anomalies is higher in the cases associated with extra-cardiac anomalies than in that with isolated CHD (64.5% versus 28.0%). In the 35 fetuses with chromosomal abnormalities, 19 (54.3%) were trisomy 18.

CONCLUSIONChromosomal abnormalities occurred in 43.2% of CHD cases and trisomy 18 is the most common aneuploidy. The likelihood of chromosomal anomaly increases when there is extracardiac involvement. Testing for the 22q11.2 microdeletion/duplication is recommended in all CHD fetuses without chromosomal anomalies. It is important for the further management of pregnancy and genetic counseling.

Adult ; Amniocentesis ; methods ; Chromosome Aberrations ; chemically induced ; classification ; Female ; Fetal Development ; genetics ; Gestational Age ; Heart Defects, Congenital ; diagnostic imaging ; genetics ; Humans ; Karyotyping ; Pregnancy ; Trisomy ; physiopathology ; Ultrasonography, Prenatal