Disruption of Planar Cell Polarity Pathway Attributable to Valproic Acid-Induced Congenital Heart Disease through Hdac3 Participation in Mice / 中华医学杂志(英文版)

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 0.05); VPA exposure dramatically decreased the expression of Vanlg2/Scrib together with Hdac activity (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.

Diagnostic value of of Peptest in laryngopharyngeal reflux disease / 医学研究生学报

Objective At present,there are relatively few studies on the application of pepsin in the diagnosis of laryngopha -ryngeal reflux disease(LPRD)in China.This paper aimed to evaluate the application value of Peptest in the diagnosis of LPRD by measuring saliva/sputum pepsin in patients with chronic pharyngitis. Methods From January to June 2017,35 patients with chronic pharyngitis treated in our department were selected to undertake electronic laryngoscopy and evaluated by reflux finding score(RFS) and reflux symptom index(RSI)scoring.The patients'saliva/sputum samples were collected three times for pepsin concentration de-tection(PEPTEST).The first collection was before mouthwash in the morning,the second was 1h after lunch,and the third was the time of evident symptoms.Comparative research was carried out on the above results. Results The average RSI score and RFS score were11.31±6.21 and 5.97±1.98.The pepsin content in morning saliva/sputum[26.80(0,109.80)ng/mL]was significantly lower than that at 1 h after lunch[89.00(16.0,254.8)ng/mL]and that at the time of obvious symptoms[105.70(34.3,254.8)ng/mL](P<0.05).The area under ROC of Peptest in the morning[0.759(0.602, 0.917)],1h after lunch[0.824(0.670,0.978)], at the time of ob-vious symptoms were all greater than the combination of RSI and RFS, representing significant statistical differences(P<0.05). Conclusion Compared with traditional method(a combination of RSI and RFS),Peptest has more important clinical value in the diag-nosis of LPRD.

Effect of Histone Deacetylase Inhibition on the Expression of Multidrug Resistance-associated Protein 2 in a Human Placental Trophoblast Cell Line / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>HDACs inhibition could up-regulate placental MRP2 expression in vitro, and HDAC1 was probably to be involved in this process.</p>

Perinatal integrative intervention for critical pulmonary artery valve stenosis / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the effect of different operation time to percutaneous balloon pulmonic valvuloplasty (PBPV) to critical pulmonary valve stenosis (CPS).</p><p><b>METHOD</b>Twenty-one infants (age ≤ 60 days at operating day) suffered from CPS, diagnosed by fetal echocardiogram and confirmed by echocardiography after birth, were enrolled in this case-control-study with written informed consent during April 2007 to December 2011. Of the 21 cases, 7 had prenatal diagnosis in our prenatal diagnosis center (prenatal group, Pre) and 14 were referred from other hospitals, who were divided into postpartum group A (Post A, referred within 28 days after birth) and postpartum group B (Post B, referred 29 to 60 days after birth). To Pre-group, the integrative interventional protocol was cautiously made by the consultative specialists, including intrauterine diagnosis, perinatal care and urgent PBPV soon after birth. To Post-group, emergency PBPV was preformed after the referral. Tei index of right ventricular and pressure-gradient (PG) between right ventricular and pulmonary artery were measured before and at different time points one year after PBPV.</p><p><b>RESULT</b>The values of SpO2 in Pre-group ranged from 82%-92% (86.57% ± 5.34%) under the state of continuous intravenous infusion of alprostadil. PBPV was successfully preformed within 3-6 days after birth. The values of SpO2 increased to 97.33% ± 1.15% post procedure. The values of PG pre- and post- procedure were (86.34 ± 11.77) mm Hg and (31.43 ± 8.46) mm Hg respectively. Preoperative RV Tei-index was 0.68 ± 0.05, it decreased rapidly after procedure, and recovered to normal one month after procedure. Only one case showed restenosis seven months after procedure and repeated PBPV. Fourteen referral cases (6 cases in Post A group and 8 cases in Post B group, accompanied in 1 and 3 cases with heart failure), the values of SpO2 ranged from 83%-91% under state of continuous intravenous infusion of alprostadil. And the operating time was 10-57 days after birth. The values of SpO2 recovered to normal post procedure, and heart failure alleviated. Increased preoperative RV pressure obviously decreased significantly post-procedure. And increased Tei-index declined gradually, at one-year follow-up, the value of Tei-index in Post A group recovered to normal, whereas that of Post B was (0.51 ± 0.06), compared to Pre and Post A groups, the difference was significant (P < 0.05) . One case showed restenosis nine months after procedure and repeated PBPV was performed. The hypoxic exposure durations were (4.43 ± 0.68) , (16.33 ± 4.46) , (41.25 ± 9.19) , respectively, and the difference among the three groups was significant (P < 0.05).</p><p><b>CONCLUSION</b>To the fetuses with definite prenatal diagnosis of critical pulmonary valve stenosis, preoperative general condition can be adjusted to more suitable for emergency operation. Early PBPV can achieve shorter hypoxic exposure and better recovery of right ventricular function post procedure. Perinatal integrated intervention for CPS can significantly improve the prognosis and quality of life in this patient population.</p>

Characteristics of changes in urinary NGAL, KIM-1 and IL-18 in Phytolaccae Radix-induced renal injury in rats and significance of combined detection / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the characteristics of changes in neutrophil gelatinase-associated lipocalcin (NGAL), kidney injury molecule-1 (KIM-1) and interleukin-18 (IL-18) in Phytolaccae Radix-induced kidney injury in rats and the significance of the combined detection.</p><p><b>METHOD</b>Wistar rats were divided into three groups: high and low dose (crude drug 40, 20 g x kg(-1) x d(-1)) Phytolaccae Radix decoction groups and the control group, and orally administrated with distilled water or equal volume of Phytolaccae Radix decoction for 35 consecutive days. Their blood and urine samples were collected on day 7, 14, 21, 28, 35,42. The anatomical analysis was conducted for each group. The contents of serum total protein (TP), albumin (ALB), blood urea nitrogen (BUN), creatinine (CR) and urinary TP and ALB were detected-by means of biochemical analyzer. The concentrations of urinary NGAL, KIM-1 and IL-18 were measured by enzyme-linked immunosorbent assay (ELISA). The morphological changes of renal pathology were observed by light or electron microscopy. The curve areas of various serum or urine indexes and the combined detection were compared by receiver operating characteristic curve (ROC curve).</p><p><b>RESULT</b>Rats were given Phytolaccae Radix decoction at the doses of 40, 20 g crude drug/kg daily for 35 consecutive days to induce kidney injury characterized by the degeneration of renal tubular epithelial cell and protein cast. The injury was partially reversible during the recovery period. Compared with the control group, the content of serum BUN, CR and urinary TP in each dose group mostly showed a downward trend. On day 21, the content of urinary ALB obviously increased till the end of administration. The contents of urinary NGAL, KIM-1 and IL-18 began increasing on day 7. Since day 14, high and low dose groups showed significant difference (P<0.01). The high dose group even showed notable changes during the recovery period. According to ROC analysis, the curve areas of NGAL, KIM-1 and IL-18 were 0.846, 0.837 and 0.863 (P <0.01), respectively, much higher than that of BUN and CR. The area of the combined detection was up to 0.947.</p><p><b>CONCLUSION</b>Urinary NGAL, IL-18 and KIM-1 could forecast and indicate the occurrence and development of renal injury to some degree, and show higher sensitivity and site specificity. The combined detection could further improve the test efficiency.</p>

Preconditioning of stem cells for the treatment of myocardial infarction / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>Poor stem cell survival is one of the obstacles for cell regeneration therapy post myocardial infarction (MI) and responsible for unsatisfactory therapeutic effectiveness. Various approaches to improve the status of these cells and increase cell survival have become research foci. The following article is a mini-review on the utilization of cell preconditioning for stem cell survival.</p><p><b>DATA SOURCES</b>The data used in this review were mainly from the articles in Medline and PubMed published from 1990 to 2010. The search terms included "preconditioning, stem cell and myocardial infarction".</p><p><b>STUDY SELECTION</b>Original articles and critical reviews selected were relevant to the review's theme.</p><p><b>RESULTS</b>The harsh ischemic and inflammatory microenvironment in the infarcted myocardium offers a significant challenge to the transplanted donor stem cells. Survival of stem cells following transplantation is affected by many factors, such as limited blood supply, nutritional deficiency, hypoxia, oxidative stress, and inflammation. Preconditioning methods have potent cytoprotective effects, which enables cells to maintain a "standby state" through programmed initiation of cell survival pathways.</p><p><b>CONCLUSIONS</b>The findings suggest that cell preconditioning can be used as an effective anti-apoptotic strategy and enable cells to withstand and survive the harsh environment after transplantation.</p>

Plasma endothelin level in hypertensive patients receiving standard anti-hypertensive therapy with or without statins / 中华心血管病杂志

<p><b>OBJECTIVE</b>To observe the association between plasma endothelin (ET) concentration and blood pressure level in essential hypertensive (EH) patients with or without complications and possible impact of statins on ET concentration.</p><p><b>METHODS</b>From Sep 2007 to Mar 2009, 149 patients with EH were analyzed [44 EH, 40 EH complicated by left ventricular hypertrophy (EH-LVH), 36 EH complicated by atrial fibrillation (EH-AF), and 29 EH complicated by lacunar infarction (EH-LI)], 30 healthy persons were selected as controls. EH patients were randomly divided into routine treatment group (calcium antagonists, ACEI, diuretics, beta-receptor blocker for 8 weeks) and simvastatin intervention group (routine treatment + simvastatin 40 mg/d for 8 weeks), plasma ET concentrations before and after drug intervention were measured.</p><p><b>RESULTS</b>(1) ET concentration was higher in EH group than that in control group [(71.42 +/- 6.62) pg/ml vs. (45.52 +/- 8.28) pg/ml, P < 0.01]. ET concentration was higher in EH-LVH group, EH-AF group and EH-LI group than that in EH group [(97.67 +/- 10.53) pg/ml, (102.15 +/- 12.96) pg/ml, (103.49 +/- 9.91) pg/ml vs. (71.42 +/- 6.62) pg/ml, P <0.01]. The degrees of elevated blood pressure was positively correlated with ET concentrations(all P < 0.001). (2) The left atrial diameters of EH-AF group were positively correlated with ET concentration (r = 0.684, P < 0.001). The left ventricular mass index of EH-LVH group were positively correlated with ET concentration (r = 0.545, P < 0.001). (3) The percentages of class 3 hypertension in EH-LVH group, EH-AF group and EH-LI group were higher than that in EH group (57.5%, 50.0%, 62.1% vs. 25.0%, all P < 0.05). (4) Blood pressure in class 3 hypertension patient treated with simvastatin decreased more significantly than that in routine treatment group (P < 0.05). (5) ET concentration of class 2 hypertension patient treated with simvastatin decreased significantly than that in routine treatment group (P < 0.05). ET concentrations of class 3 hypertension patient treated with simvastatin and routine treatment patient decreased significantly after treatment (P < 0.05), and the former was lower (P < 0.05).</p><p><b>CONCLUSION</b>The level of ET were positively correlated with the severity of EH. Simvastatin could decrease the ET levels of patients with EH and blood pressure levels of patients with class 3 hypertension. It suggested that therapeutic alliance of antihypertensive drugs and statins could be benefit to patients with EH.</p>