[Effect of histone acetylation/deacetylation imbalances on key gene of planar cell polarity pathway].

OBJECTIVE: To investigate the effect of histone acetylation/deacetylation imbalances on embryonic hearts of mice and its effect on key genes of planar cell polarity (PCP) pathway-Vangl2, Scrib and Rac1 in H9C2 cells. METHODS: Forty pregnant C57/B6 mice were randomly assigned into three groups: blank group (n=10), vehicle group (n=10), and valproic acid (VPA)-treated group (n=20). In the VPA-treated group, VPA, a histone deacetylase (HDAC) inhibitor, was administered to each individual dam intraperitoneally at a single dose of 700 mg/kg on embryonic day 10.5 (E10.5). The vehicle and blank groups received equivalent saline or no interventions, respectively. Dams were sacrificed on E15.5, and death rates of embryos were evaluated. Subsequently, embryonic hearts of survival fetus were removed to observe cardiac abnormalities by hematoxylin-eosin (HE) staining. H9C2 cells were cultured and allotted to the blank, vehicle, and VPA-treated groups: the VPA treated group received VPA exposure at concentrations of 2.0, 4.0 and 8.0 mmol/L; the vehicle and blank groups received equivalent saline or no interventions, respectively. HDAC1-3 as well as Vangl2, Scrib and Rac1 mRNA and protein expression levels were determined by quantitative real-time PCR and Western blot, respectively. The total HDAC activity was analyzed by colorimetric assay. RESULTS: The fetus mortality rate after VPA treatment was 31.7%, with a significantly higher rate of cardiac abnormalities in comparison with the controls (P<0.05). In comparison with the blank and vehicle groups, HDAC1 mRNA was significantly increased at various concentrations of VPA treatment at all time points of exposure (P<0.05), together with a reduction of protein level after 48 and 72 hours of exposure (P<0.05). The inhibition of HDAC2 mRNA after various concentrations of VPA incubation was pronounced at 24 hours of exposure (P<0.05), while the protein levels were reduced at all time points (P<0.05). HDAC3 mRNA was prominently induced by VPA (4.0 and 8.0 mmol/L) at all time points of treatment (P<0.05). In contrast, the protein level was inhibited after VPA treatment (P<0.05). In comparison with the blank and vehicle groups, Vangl2 mRNA as well as Scrib mRNA/protein expression levels were markedly reduced after 48 and 72 hours of VPA treatment (P<0.05), together with a reduction of protein level in Vangl2 at 72 hours (P<0.05). Compared with the blank and vehicle groups, a significant repression in the total HDAC activity was observed in the VPA-treated group at concentrations of 4.0 and 8.0 mmol/L after 24 hours of treatment (P<0.05), and the effect persisted up to 48 and 72 hours, exhibiting pronounced inhibition at all concentrations (P<0.05). CONCLUSIONS: VPA might result in acetylation/deacetylation imbalances by inhibiting HDAC1-3 protein expression and total HDAC activity, leading to the down-regulation of mRNA and protein expression of Vangl2 and Scrib. This could be one of the mechanisms contributing to congenital heart disease.

[Changes in electrocardiographic parameters in children with complete left bundle branch block early after transcatheter closure of ventricular septal defect].

OBJECTIVE: To observe the changes in electrocardiographic parameters in children with complete left bundle branch block (CLBBB) after the transcatheter closure of simple ventricular septal defect (VSD). METHODS: A total of 21 children with CLBBB early after transcatheter closure of perimembranous VSD were recruited. Another 21 children without any type of arrhythmia after transcatheter closure of perimembranous VSD were enrolled as the control group. The sex, age, and the size of occluder were matched between the two groups. The changes in the following indices were compared between the two groups: left ventricular voltage, QT interval, corrected QT interval (QTc), QT dispersion (QTd), corrected QT dispersion (QTcd), JT dispersion (JTd), and corrected JT dispersion (JTcd) on the electrocardiogram before transcatheter closure and at 1, 3, 5, 30 days after transcatheter closure. RESULTS: Left ventricular voltage and JTcd changed with operation time in the CLBBB and control groups (P<0.05). There were interaction effects between time and grouping in the changes in left ventricular voltage and QTd (P<0.05). There was a significant difference in JTcd between the CLBBB and control groups (P<0.05). There was also a significant difference in left ventricular voltage between the CLBBB and control groups at 3 and 5 days after the transcatheter closure (P<0.05). CONCLUSIONS: There are significant differences in electrocardiographic left ventricular voltage and JTcd between VSD children with and without CLBBB early after transcatheter closure. JTcd might be useful in predicting the development of CLBBB early after transcatheter closure of VSD.

[Research advances on role of microRNAs in congenital heart diseases].

MicroRNAs (miRNAs) are a class of small non-coding RNAs, which mainly regulate gene expression through post-transcriptional process. They are highly conserved, tissue-specific and highly specific in miRNA-binding on 3'-untranslated regions. MicroRNAs have been identified as crucial regulators in myocardial cell proliferation, differentiation and apoptosis, migration of cardiac neural crest cells, cardiac morphogenesis and cardiac patterning processes, which may provide a new insight into the research on developmental mechanism of congenital heart diseases. The research on miRNAs in congenital heart diseases includes clinical research and animal experiments. This article reviews two types of research advances, the mechanism of congenital heart diseases, and the current status and limitation of the domestic reports.

[Significance of Tp-Te interval for risk stratification of ventricular premature contractions in children].

OBJECTIVE: To investigate the significance of Tp-Te interval for risk stratification of ventricular premature contractions (VPC) in children. METHODS: A total of 120 children with VPC were divided into benign VPC (n=40), organic disease (n=40) and ventricular parasystole groups (n=40) according to the etiology of VPC; another 40 children who underwent physical examination were selected as the normal control group. The four groups were compared in terms of Tp-Te intervals and Tp-Te/QT ratios in leads V3, V4 and V5. RESULTS: The Tp-Te interval in lead V3 was significantly longer in the organic disease group than in the other groups (P<0.05), the benign VPC group had a significantly shorter Tp-Te interval in lead V4 than the normal control and organic disease groups (P<0.05), and the organic disease group had a significantly longer Tp-Te interval in lead V5 than the benign VPC group (P<0.05). The Tp-Te/QT ratios in leads V3-V5 were significantly higher in the organic disease group than in the other groups (P<0.05). The Tp-Te/QT ratios in leads V4 and V5 showed significant differences between the ventricular parasystole and benign VPC groups (P<0.05). CONCLUSIONS: Tp-Te interval is susceptible to changes in heart rate, and it is of little value for the risk stratification of VPC in children. Tp-Te/QT ratio, however, may be used as an important non-invasive index for clinical risk stratification of VPC in children and is worthy of further study.

[Risk factors for accelerated junctional escape rhythm in children early after percutaneous ventricular septal defect closure].

OBJECTIVE: To identify the risk factors for accelerated junctional escape rhythm (AJER) in children early after percutaneous ventricular septal defect (VSD) closure. METHODS: A retrospective controlled study was conducted on 42 children who had AJER within one week after percutaneous VSD closure between January 2008 and October 2012. These subjects were compared with controls without AJER after VSD closure in terms of age, sex, diameter of VSD, occluder size, difference between occluder size and diameter of VSD, and distance between VSD and aortic valve ring. Risk factors for AJER were identified by logistic regression analysis. RESULTS: Compared with the control group, the AJER group had a longer distance betweenVSD and aortic valve ring, a larger diameter of VSD (basal diameter), a larger occluder size (waist diameter) , and a bigger difference between the waist diameter of occluder and diameter of VSD (P<0.05). Logistic regression analysis showed that distance between VSD and aortic valve ring (OR=1.813, P<0.05) and occluder size (OR=1.671, P<0.05) are primary risk factors for AJER. CONCLUSIONS: AJER early after percutaneous VSD closure is related to diameter of VSD, occluder size, difference between the waist diameter of occluder and diameter of VSD, and distance between VSD and aortic valve ring. The distance between VSD and aortic valve ring and occluder size are primary risk factors for AJER.

[The influence of inhibiting notch signal on pulmonary vascular remodeling induced by PDGF].

OBJECTIVE: To explore the influence of inhibiting Notch signal on pulmonary vascular remodeling induced by PDGF. METHODS: Vessel strips taken from healthy Wistar rats were cultured together with extrogenous PDGF, the potent smooth muscle cell proliferation stimulators, for 7 days. Some vessel strips were cultured with PDGF and gamma-secretase inhibitor DAPT, a Notch signaling inhibitor for 7 days. Vascular wall thickness, PCNA and caspase-3 positive cell rate were examined in vessel strips. The alterations of Notch 1 to 4 receptor and HERP1, 2 mRNA were discerned by FQ-PCR to observe the influence of DAPT on Notch signal. At the same time, above indexes, which were related with pulmonary vascular remodeling, were measured too. RESULTS: PDGF stimulation in the cultured normal pulmonary arteries resulted in vascular medial thickness increase for about 50%, accompanied by significant increase in PCNA positive cell rate and decrease of caspase-3 positive cell rate. When DAPT were added to inhibit Notch signaling, the expression of HERP1, 2 mRNA decreased, the degrees of PDGF induced vascular medial thickness and PCNA positive cell rate increase as well as caspase-3 positive cell rate decrease were all attenuated notably. CONCLUSION: Inhibiting Notch signal induced by gamma-secretase inhibitor lead to the suppression of pulmonary vascular remodeling induced by PDGF, suggesting inhibition of Notch signal pathway might be a novel strategy in the intervention of pulmonary hypertension.

[Effect of inhibition of Notch signal on pulmonary vascular remodeling induced by angiotensin â ¡].

OBJECTIVE: It is known that Notch signal is very important to vascular remodeling during the process of embryonic development, vessel repair and tumor growth, but there are few studies about pulmonary vascular remodeling in pulmonary hypertension. This study was to explore the effect of inhibiting Notch signal on pulmonary vascular remodeling induced by angiotensin II. METHODS: Vessel strips taken from healthy Wistar rats were co-cultured with extrogenous angiotensin II and the potent smooth muscle cell proliferation stimulators for 7 days. Vascular wall thickness, proliferating cell nuclear antigen (PCNA) positive cell rate and caspase-3 positive cell rate were examined in vessel strips. Then some vessel strips were cultured with angiotensin II and γ-secretase inhibitor DAPT, a Notch signaling inhibitor for 7 days. The levels of Notch 1 to 4 receptor and HERP1/2 mRNA were ascertained by FQ-PCR. RESULTS: Angiotensin II stimulation in the cultured normal pulmonary arteries resulted in an increase in the vascular medial thickness by nearly 50%, and a significant increase in the PCNA positive cell rate and a decrease in the caspase-3 positive cell rate. DAPT treatment did not result in the alterations of Notch 1 to 4 receptor levels, but decreased remarkably HERP1 and HERP2 mRNA expression. DAPT treatment also decreased angiotensin II-induced vascular medial thickness and PCNA positive cell rate and increased caspase-3 positive cell rate. CONCLUSIONS: Inhibiting Notch signal by γ-secretase inhibitor may lead to the suppression of pulmonary vascular remodeling induced by angiotensin II, suggesting that the inhibition of Notch signal pathway might be a novel strategy for the treatment of pulmonary hypertension.

[Echocardiography diagnosis of fetal cardiac structural abnormalities].

OBJECTIVE: To investigate the application of echocardiography diagnosis of fetal cardiac structural abnormalities. METHODS: The echocardiography findings of 9 352 fetus were studied. RESULTS: A total of 472 cases showed cardiac structural abnormalities, including 7 cases of ventricular septal defect, 53 cases of atrioventricular septal defect, 49 cases of atrial septal defect, 26 cases of tetralogy of Fallot, 21 cases of persistent truncus arteriosus, 20 cases of Ebstein's anomaly and 206 cases of other cardiac abnormalities. There were 17 cases with cardiac arrhythmia, 9 with heart failure, and 5 with hydrops. CONCLUSIONS: Fetal echocardiography is a promising diagnostic tool for prenatal evaluation of cardiac structural abnormalities. The echocardiography diagnosis and evaluation for fetal congenital cardiovascular malformations is the foundation of the guidance and monitoring in intrauterine fetal cardiac intervention.

Incomplete Kawasaki disease complicated with acute abdomen: A case report.

BACKGROUND: Kawasaki disease (KD) is an acute systemic vasculitis characterized by unknown etiology. CASE SUMMARY: A 4.5-year-old boy developed an acute abdomen during the onset of incomplete KD. He still had persistent abdominal pain after undergoing exploratory laparotomy and appendectomy. Ultrasound examination at early onset revealed a giant coronary artery aneurysm. The patient developed a myocardial infarction and heart failure accompanied by respiratory and cardiac arrest. He underwent coronary artery revascularization and coronary artery bypass graft using an autologous internal mammary artery. After the operation, the cardiac output increased, and the symptoms of heart failure resolved. Follow-up evaluation at 1 mo after operation showed that the patient's cardiac function had restored to New York Heart Association standard Grade I heart failure, and normal growth was obtained. CONCLUSION: Coronary artery revascularization and coronary artery bypass graft is an effective method for treating myocardial ischemia in children with KD complicated with giant coronary artery aneurysm . Nevertheless, some issues still need specific attention.

[Changes of MMP-2,9 and TIMP-1 expressions in rats with pulmonary arterial hypertension after captopril and losartan interventions].

OBJECTIVE: To determine the effect of captopril and losartan on the expressions of matrix metalloproteinase-2,9 (MMP-2,9) and metalloproteinase-1 (TIMP-1) in rats with pulmonary arterial hypertension, and the mechanisms of captopril and losartan in intervening the development of pulmonary arterial hypertension. METHODS: Forty male Spraque-Dawley rats were divided into 4 groups randomly: pulmonary arterial hypertension (created by pneumonectomy plus MCT injection) model group (PAH Model), PAH model treated with captopril [PAH+Cap 10 mg/(kg x d)], losartan group [PAH+Los 15 mg/(kg x d)] and normal control group(Control). The mPAP, weight ratio of RV to LV+S, neointima formation, relative thickness of small pulmonary arteries, and degree of muscularization of non-muscular arterioles were measured at day 35. The expression of SM-a-actin in the PASMC was determined by immunochemistry stain. The expressions of MMP-2, 9, TIMP-1 and MMP-2, 9, TIMP-1 mRNA in the pulmonary tissues were determined by immunohistochemistry and FQ-PCR respectively. The enzymatic activity of MMP-2, 9 was measured by Gelatin zymography. RESULTS: Pneumonectomy plus MCT injection induced severe pulmonary arterial hypertension characterized by neointimal formation. Captopril or losartan suppressed the increase of mPAP, right ventricle weight, thickness of small pulmonary arteries and muscularization of peripheral pulmonary arterioles in the rats with PAH (P < 0.05). The PAH model group had higher expressions of MMP-2, 9, TIMP-1 mRNA and enzymatic activity of MMP-2, 9 in lung tissue than the other groups (P < 0.05). Captopril intervention had similar effects as losartan intervention. CONCLUSION: The captopril and losartan induced attenuation of PAH and pulmonary vascular remodeling is likely to be associated with the regulation of the expressions of MMP-2, 9, TIMP-1.

[Effect of polycythemia on hypoxia induced pulmonary hypertension and pulmonary vascular remodeling in rats].

OBJECTIVE: To investigate the effect of polycythemia on hypoxia induced pulmonary hypertension and pulmonary vascular remodeling in rats. METHODS: The healthy female Sprague-Dawley rats were randomly divided into 3 groups: normoxia control group (C group), hypoxia group (H group), hypoxia + different doses of human recombine hemopoietin (rEPO) group. All rats in hyoxia groups were exposed to hypoxia, 8 hours every day, for 21 days. The rEPO groups were injected sc with different doses of rEPO (300 U/kg, 600 U/kg, 900 U/kg, 1200 U/kg) thrice weekly. Blood samples were taken for the measurement of RBC, Hb, Hct, plasma EPO concentration, whole blood/plasma viscosities, the animals were then catheterized to record mean pulmonary arterial pressure (mPAP) and demised to calculate the ratio [RV/(LV+S)]. Percentage of vascular wall thickness and muscularization of non-muscular pulmonary arteriole were examined microscopically. RESULTS: (1) As the dosage of exogenous rEPO increased, blood concentration of EPO increased correspondingly, as RBC, Hb, Hct and whole blood/plasma viscosities increased in various degrees. (2) There was positive correlation between whole blood viscosity and Hct at both high and low shears and linear correlation between mPAP and whole blood viscosity at high shear. (3) The degree of pulmonary hypertension, reflected by mPAP increased in accordance to rEPO dosage increment. However, the extent of pulmonary vascular remodeling alleviated somehow as the rEPO dose increased and so did right ventricular hypertrophy. CONCLUSION: Polycythemia induced by exogenous EPO increases the blood viscosity and the pulmonary vascular resistance, which contributes to the formation of hypoxia induced pulmonary hypertension.

[Effect of resveratrol on myocardial fibrosis in mice with chronic viral myocarditis].

OBJECTIVE: Some research has shown that resveratrol can ameliorate myocardial injury and improve cardiac function in mice with acute viral myocarditis (VMC), and can inhibit cardiac fibroblast proliferation and myofibroblast differentiation in vitro. This study was designed to investigate whether resveratrol has similar effects in the mouse model of chronic VMC. METHODS: One hundred mice were inoculated with 0.3 mL of Coxsackievirus B3 1*106 TCID50. Thirty days later, the survivors (n=62) were used as a model of chronic VMC, and were randomly assigned to 4 groups: untreated VMC, and low- (10 mg/kg), middle- (100 mg/kg) and high-dose (1 000 mg/kg) resveratrol-treated VMC (once daily, for 30 days). Ten mice which received neither Coxsackievirus B3 nor resveratrol treatment served as the control group. After 30 days of resveratrol treatment, the mice were sacrificed. Serum concentrations of collagenous pre-peptides (PINP, PICP and PIIINP) were assessed using ELISA. Hematoxylin-eosin staining, picrosirius red staining and circularly polarized light were used to examine the histochemistry of myocardial collagen. RESULTS: The myocardial collagen volume fraction in the high-dose (0.74+/-0.19) and the middle-dose (1.07+/-0.12) resveratrol-treated VMC groups was significantly lower than that in the untreated VMC (2.33+/-0.18) and the low-dose resveratrol-treated VMC (2.17+/-0.19) groups (P<0.05). Compared with the untreated VMC group, serum concentrations of PICP and PIIINP in the high-dose and the middle-dose resveratrol-treated VMC groups were significantly reduced (P<0.05), while PINP concentrations increased significantly (P<0.05). CONCLUSIONS: Resveratrol can inhibit hyperplasia of myocardial collagen in the mouse model of chronic VMC, acting as an effective anti-fibrotic agent in the myocardium.

[The research of animal model setting-up in human intra-utero fetal cardiac intervention].

OBJECTIVE: To research the techniques and correlated issues during analogous procedure of human intro-utero fetal cardiac intervention in animal model, so as to impulse the clinical utilizing of cardiac intervention in human fetus with severe cardiovascular malformations. METHODS: Eight bigeminal pregnant ewes in latter 2nd-trimester and 3rd- trimester were taken into the research of fetal cardiac intervention. Under continuous ultrasound guidance, advanced an cannula and stylet needle through the maternal abdomen, uterine wall, and fetal lamb chest wall and into the fetal LV, then imitate human balloon aortic valvuloplasty at valve ring level. Probed into aspects of animal preparation, position of fetal lambs, paracentesis point selecting, main points of intervention, ultrasound utilizing, fetal lambs incubation and protection to placenta as well as umbilical cord. RESULTS: Eight pregnant ewes were all survival after procedure, 2 was executed after fetus' death, the other 6 continued gestation until spontaneous vaginal delivery after an uneventful pregnancy. After parturition the 6 ewes were in good condition. There were no nick infection, chorioamnionitis and other complications. Eight/sixteen fetal lambs were undergone intro-utero cardiac intervention. The values of body weight and Hct of lambs which were executed, pre-term and full-term delived were (1.77 +/- 0.14) kg vs. (2.15 +/- 0.23) kg vs. (2.41 +/- 0.19) kg and 29%o-33% vs. 27%-35% vs. 37%-41%. In autopsy, hydropericardium was found in 4/8 with 1-2.5 mL and 1/8 with 5 mL. Besides 1/8 with interventricular septum centesis injury, there was no centesis damage to endocardium, valve, chordae tendineae, papillary muscles and sortic tunics intima. Whereas there was no centesis injury to lung, liver and chylostomach, no trace of ericardium and thoracic infection. CONCLUSIONS: Animal model of intra-utero fetal cardiac intervention was setted up successfully, the experiences in this study was worth to be payed attention to in human fetal cardiac intervention.

Disruption of Planar Cell Polarity Pathway Attributable to Valproic Acid-Induced Congenital Heart Disease through Hdac3 Participation in Mice.

BACKGROUND: Valproic acid (VPA) exposure during pregnancy has been proven to contribute to congenital heart disease (CHD). Our previous findings implied that disruption of planar cell polarity (PCP) signaling pathway in cardiomyocytes might be a factor for the cardiac teratogenesis of VPA. In addition, the teratogenic ability of VPA is positively correlated to its histone deacetylase (HDAC) inhibition activity. This study aimed to investigate the effect of the VPA on cardiac morphogenesis, HDAC1/2/3, and PCP key genes (Vangl2/Scrib/Rac1), subsequently screening out the specific HDACs regulating PCP pathway. METHODS: VPA was administered to pregnant C57BL mice at 700 mg/kg intraperitoneally on embryonic day 10.5. Dams were sacrificed on E15.5, and death/absorption rates of embryos were evaluated. Embryonic hearts were observed by hematoxylin-eosin staining to identify cardiac abnormalities. H9C2 cells (undifferentiated rat cardiomyoblasts) were transfected with Hdac1/2/3 specific small interfering RNA (siRNA). Based on the results of siRNA transfection, cells were transfected with Hdac3 expression plasmid and subsequently mock-treated or treated with 8.0 mmol/L VPA. Hdac1/2/3 as well as Vangl2/Scrib/Rac1 mRNA and protein levels were determined by real-time quantitative polymerase chain reaction and Western blotting, respectively. Total HDAC activity was detected by colorimetric assay. RESULTS: VPA could induce CHD (P < 0.001) and inhibit mRNA or protein expression of Hdac1/2/3 as well as Vangl2/Scrib in fetal hearts, in association with total Hdac activity repression (all P < 0.05). In vitro, Hdac3 inhibition could significantly decrease Vangl2/Scrib expression (P < 0.01), while knockdown of Hdac1/2 had no influence (P > 0.05); VPA exposure dramatically decreased the expression of Vanlg2/Scrib together with Hdac activity (P < 0.01), while overexpression of Hdac3 could rescue the VPA-induced inhibition (P > 0.05). CONCLUSION: VPA could inhibit Hdac1/2/3, Vangl2/Scrib, or total Hdac activity both in vitro and in vivo and Hdac3 might participate in the process of VPA-induced cardiac developmental anomalies.

Autoimmune-associated Congenital Heart Block: A New Insight in Fetal Life.

OBJECTIVE: Congenital heart block (CHB) is a rare but life-threatening disorder. More than half of CHB cases are associated with maternal autoimmune, which are termed as autoimmune-associated CHB. This review summarized the recent research findings in understanding autoimmune-associated CHB, discussed the current diagnostic approaches and management strategies, and summarized the problems and future directions for this disorder. DATA SOURCES: We retrieved the articles published in English from the PubMed database up to January 2017, using the keywords including "Autoimmune-associated", "Autoimmune-mediated", and "Congenital heart block". STUDY SELECTION: Articles about autoimmune-associated CHB were obtained and reviewed. RESULTS: Observational studies consistently reported that transplacental maternal antibodies might recognize fetal or neonatal antigens in various tissues and result in immunological damages, but the molecular mechanisms underlying CHB pathogenesis still need illuminated. Multiple factors were involved in the process of atrioventricular block development and progression. While several susceptibility genes had been successfully defined, how these genes and their protein interact and impact each other remains to be explored. With currently available diagnostic tools, fetal ultrasound cardiography, and fetal magnetocardiography, most of CHB could be successfully diagnosed and comprehensively evaluated prenatally. The efficacy of current approaches for preventing the progression and recurrence of CHB and other autoimmune-mediated damages was still controversial. CONCLUSIONS: This review highlighted the relationships between autoimmune injuries and CHB and strengthened the importance of perinatal management and therapy for autoimmune-associated CHB.

Effect of Histone Deacetylase Inhibition on the Expression of Multidrug Resistance-associated Protein 2 in a Human Placental Trophoblast Cell Line.

BACKGROUND: Placental multidrug resistance-associated protein 2 (MRP2), encoded by ABCC2 gene in human, plays a significant role in regulating drugs' transplacental transfer rates. Studies on placental MRP2 regulation could provide more therapeutic targets for individualized and safe pharmacotherapy during pregnancy. Currently, the roles of epigenetic mechanisms in regulating placental drug transporters are still unclear. This study aimed to investigate the effect of histone deacetylases (HDACs) inhibition on MRP2 expression in the placental trophoblast cell line and to explore whether HDAC1/2/3 are preliminarily involved in this process. METHODS: The human choriocarcinoma-derived trophoblast cell line (Bewo cells) was treated with the HDAC inhibitors-trichostatin A (TSA) at different concentration gradients of 0.5, 1.0, 3.0, and 5.0 µmol/L. Cells were harvested after 24 and 48 h treatment. Small interfering RNA (siRNA) specific for HDAC1/HDAC2/HDAC3 or control siRNA was transfected into cells. Total HDAC activity was detected by colorimetric assay kits. HDAC1/2/3/ABCC2 messenger RNA (mRNA) and protein expressions were determined by real-time quantitative polymerase chain reaction and Western-blot analysis, respectively. Immunofluorescence for MRP2 protein expression was visualized and assessed using an immunofluorescence microscopy and ImageJ software, respectively. RESULTS: TSA could inhibit total HDAC activity and HDAC1/2/3 expression in company with increase of MRP2 expression in Bewo cells. Reduction of HDAC1 protein level was noted after 24 h of TSA incubation at 1.0, 3.0, and 5.0 µmol/L (vs. vehicle group, all P < 0.001), accompanied with dose-dependent induction of MRP2 expression (P = 0.045 for 1.0 µmol/L, P = 0.001 for 3.0 µmol/L, and P < 0.001 for 5.0 µmol/L), whereas no significant differences in MRP2 expression were noted after HDAC2/3 silencing. Fluorescent micrograph images of MRP2 protein were expressed on the cell membrane. The fluorescent intensities of MRP2 in the control, HDAC2, and HDAC3 siRNA-transfected cells were week, and no significant differences were noticed among these three groups (all P > 0.05). However, MRP2 expression was remarkably elevated in HDAC1 siRNA-transfected cells, which displayed an almost 3.19-fold changes in comparison with the control siRNA-transfected cells (P < 0.001). CONCLUSIONS: HDACs inhibition could up-regulate placental MRP2 expression in vitro, and HDAC1 was probably to be involved in this process.

[The analysis of T cell receptor Vbeta gene usage of cytolytic T-lymphocytes in coxsackievirus B3-induced myocarditis].

OBJECTIVE: To analyze the T cell receptor (TCR) Vbeta gene usage of the cytolytic T-lymphocytes which result in immune-mediated myocyte injury in Coxsackievirus B3-induced myocarditis. METHODS: An experimental murine model with a myocarditic variant of Coxsackievirus B, type 3 has been developed with newborn Balb/c male mice. Mice were killed on day 7 after virus inoculated, which was the time when T cells show significant differentiation. With immunoadsorption, ACTLs, VCTLs, MCTLs are respectively isolated from mesenteric lymph nodes of Balb/c mice suffered from viral myocarditis. T-lymphocytes prepared from mesenteric lymph nodes of uninfected Balb/c mice served as control group. RT-PCR was performed with standard method. The Vbeta gene usage was analyzed by running the electophoresis of PCR products on agarose gel. RESULTS: T cells of control group expressed all Vbeta genes of 20 families. But the TCR VP gene usage of CTLs in experimental groups was markedly restricted. ACTL expressed predominantly Vbeta6, Vbeta8.1, Vbeta8.2, Vbeta8.3, MCTL expressed predominantly Vbeta5.1, Vbeta8.1, Vbeta8.2, Vbeta8.3 and VCTL expressed predominantly Vbeta7, Vbeta8.1, Vbeta8.2, Vbeta8.3. CONCLUSIONS: The TCR Vbeta gene usage of CTLs which result in immune-mediated myocyte injury in Coxsackievirus B3-induced myocarditis is markedly restricted in cells stimulated by specific antigen.

Efficacy of prenatal diagnosis of major congenital heart disease on perinatal management and perioperative mortality: a meta-analysis.

BACKGROUND: There is no consensus on the effectiveness of prenatal diagnosis except for hospitalized outcomes. Hence, a meta-analysis of published literature was conducted to assess the effect of prenatal diagnosis. METHODS: Literature review has identified relevant studies up to December 2013. A meta-analysis was performed according to the guidelines from the Cochrane review group and the PRISMA statement. Studies were identified by searching PubMed, Embase, the Cochrane Central Register of Controlled Trials and World Health Orgnization clinical trials registry center. Meta-analysis was performed in a fixed/random-effect model using Revman 5.1.1 according to the guidelines from the Cochrane review group and the PRISMA guidelines. RESULTS: The results from 13 cohort studies in 12 articles were analyzed to determine the optimal treatment with the lower rate of perioperative mortality in prenatal diagnosis. The superiority of a prenatal diagnosis has been proven because the surgical procedure could be done in the early neonatal period (95% CI, -0.76, -0.40). The prenatal diagnosis has also remarkably reduced the preoperative and postoperative mortality rates in cases of transposition of the great arteries (95% CI=0.06, 0.80; 95% CI=0.01, 0.82, respectively), as well as the overall results with all subtypes (95% CI=0.18, 0.94; 95% CI=0.46, 0.94, respectively). CONCLUSIONS: Prenatal diagnosis is effective in perinatal management with an earlier intervention for major congenital heart disease, but only results in a reduced perioperative mortality in cases of transposition of the great arteries. Further investigations are required to evaluate the effect of prenatal diagnosis on life quality during a long-term follow-up.