Blue-emitting tryptophan-protected gold nanoclusters acted as a sensitive nanosensor for fluorescence sensing and visual imaging detection of furaltadone.

Herein, blue-emitting gold nanoclusters (Au NCs) were carried out through tryptophan as the protecting and reducing agents. In aqueous solution of Au NCs@tryptophan, the addition of furaltadone guaranteed the interaction of furaltadone with tryptophan around Au NCs. The propinquity of furaltadone to Au NCs caused that the fluorescence of Au NCs was weakened by furaltadone based on the inner filter effect (IFE). Under the optimal measurement conditions, the logarithm of relative fluorescence intensity of Au NCs@tryptophan was linearly carried out with the furaltadone amount increasing from 0.5 to 100 µM, the corresponding detection limit was 0.087 µM. The fluorescence change of Au NCs@tryptophan displayed excellent selectivity and sensitivity for furaltadone than other possible substance in the human body. In view of Au NCs@tryptophan, the as-performed fluorescence nanosensor suggested outstanding ability for furaltadone sensing in real samples. Obviously, this nanoprobe of furaltadone could implement the naked-eye visual fluorescence determination of furaltadone.

Preparation of blue fluorescent copper nanoclusters for sensitive and selective sensing of apigenin in pharmaceutical samples.

One-pot means was performed for the rapid preparation of copper nanoclusters (Cu NCs), which were employed as a fluorescence system for the sensitive apigenin measurement in pharmaceutical samples. Herein, CuCl2 aqueous solution was reduced to Cu NCs through ascorbic acid and the Cu NCs were protected through trypsin under 65 ℃ for 4 h. The entire preparation process was rapid, facile and environmentally friendly. The trypsin-capped Cu NCs were demonstrated through ultraviolet-visible spectroscopy, fluorescence spectroscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy and fluorescence lifetime, respectively. The Cu NCs revealed blue fluorescence with emission wavelength around 465 nm under the excitation wavelength of 380 nm. The fluorescence weakening feature of Cu NCs with apigenin was observed. On this basis, a facile and sensitive turn-off fluorescent nanoprobe for the sensing of apigenin in real samples was developed. The logarithm of relative fluorescence intensity revealed a good linear relationship with apigenin contents from 0.5 µM to 300 µM with the detection limit of 0.079 µM. The Cu NCs-based fluorescent nanosensor have been employed to measure the apigenin amounts in real samples such as medical saline, bovine and human serum. The results revealed excellent potential of this Cu NCs-based fluorescent nanoprobe for the convention computation of apigenin amounts in real samples.

Selective determination of ellagic acid in aqueous solution using blue-green emissive copper nanoclusters.

Development of beneficial sensors to analyze ellagic acid concentrations is of great importance for food safety and human health. Herein, a facile and fast fluorescent probe was carried out for the excellently selective and sensitive measurement of ellagic acid in real samples through histidine protected copper nanoclusters (histidine@Cu NCs) as a nanosensor. This as-developed histidine@Cu NCs were performed through UV-vis absorption spectroscopy, fluorescence spectroscopy, transmission electron microscopy, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy and fluorescence lifetime analysis. The TEM image revealed that this nanomaterial had spherical features with the average diameter of 2.5 ± 0.05 nm. The blue-green fluorescence of this Cu NCs was found under the UV light. Meanwhile, the maximum excitation and emission wavelength were located at 387 nm and 488 nm. After addition of ellagic acid, the fluorescence of histidine@Cu NCs was slowly weakened with excellent linear range of 0.5-300 µM and detection limit of 0.077 µM. The fluorescence weakening mechanism of this nanosensor were attributed to the inner filter effect (IFE) and static quenching. Finally, this as-established analysis platform was successfully employed to measure ellagic acid in real samples.

Fluorescent folic acid-capped copper nanoclusters for the determination of rifampicin based on inner filter effect.

Development of excellent sensors to determine trace concentrations of rifampicin is of intense importance for medicine analysis and human health. Herein, a facile and green fluorescent probe was established for the determination of rifampicin by using folic acid protected copper nanoclusters (FA-Cu NCs). Many characterization methods were applied for the analysis of the as-prepared FA-Cu NCs including UV-visible absorption spectra, fluorescence spectra, Fourier-transform infrared spectroscopy (FT-IR), transmission electron microscope (TEM), fluorescence lifetime and X-ray photoelectron spectroscopy (XPS). The TEM image suggested that the as-prepared FA-Cu NCs were highly dispersed. The as-synthesized FA-Cu NCs emerged blue fluorescence under UV light and demonstrated maximum emission wavelength at 446 nm under the maximum excitation wavelength of 358 nm. After the addition of rifampicin, the FL intensities of FA-Cu NCs were uncommonly quenched. The related experimental data intimated that the quenching mechanisms were assumed to the inner filter effect (IFE) and static quenching. The as-proposed probe platform displayed an obvious linear relationship with rifampicin concentrations varying from 0.5 to 100 µM, and the corresponding detection limit (LOD) was 0.073 µM (S/N = 3). Finally, the as-established detection platform was successfully employed to analyze trace concentrations of rifampicin in real samples.

One-step synthesis of blue emission copper nanoclusters for the detection of furaltadone and temperature.

Polyvinyl pyrrolidone (PVP), playing roles as a templating agent, can be applied to prepare blue-emitting copper nanoclusters (Cu NCs@PVP) on the basis of a rapid chemical reduction synthesis method. The Cu NCs@PVP displayed a blue emission wavelength at 430 nm and the corresponding quantum yield (QY) could reach 10.4%. Subsequently, the as-synthesized Cu NCs@PVP were used for the trace analysis of furaltadone based on the inner filter effect (IFE) between Cu NCs@PVP and furaltadone, which caused the fluorescence to be effectively quenched. Additionally, this proposed determination platform based on the Cu NCs@PVP for furaltadone sensing possessed an excellent linear range from 0.5 to 100 µM with a lower detection limit of 0.045 µM (S/N = 3). Meanwhile, the Cu NCs@PVP also could be applied for the sensing of temperature. Furthermore, the practicability of the sensing platform has been successfully verified by measuring furaltadone in real samples, affirming its potential to increase fields for the determination of furaltadone.

The distinct role of orbitofrontal and medial prefrontal cortex in encoding impulsive choices in an animal model of attention deficit hyperactivity disorder.

Attention deficit hyperactivity disorder (ADHD) is a complex neurodevelopmental disorder affecting up to 5% of children worldwide. The lack of understanding of ADHD etiology prevented the development of effective treatment for the disease. Here, using in vivo electrophysiology recordings, we have recorded and analyzed the neuronal encoding of delay discounting behavior in prefrontal and orbitofrontal cortex of spontaneously hypertensive rat (SHR). We found that in the presence of rewards, neurons in the orbitofrontal cortex (OFC) were activated regardless to the value of the rewards and OFC neurons in SHR exhibited significantly higher rates of neuronal discharging towards the presence of rewards. While in the medial prefrontal cortex (mPFC), neurons of SHR responded similarly in the presence of large rewards compared with control rats whereas they displayed higher firing rates towards smaller rewards. In addition, the reward-predicting neurons in the OFC encodes for value of rewards in control animals and they were strongly activated upon receiving a small immediate reinforcer in the SHR whereas the reward-predicting neurons in the mPFC neurons generally did not respond to the value of the rewards. Our study characterized the neuronal discharging patterns of OFC and mPFC neurons in the SHR and the control animals and provided novel insights for further understanding the neuronal basis of ADHD pathology.

One facile fluorescence strategy for sensitive determination of baicalein using trypsin-templated copper nanoclusters.

Herein, we established a fluorescent detection platform for baicalein (Bai) based on copper nanoclusters, which were prepared by using copper sulfate as the precursor, trypsin (Tryp) as the template and hydrazine hydrate as the reducing agent. The entire preparation and testing process were rapid, facile and green. Many characterization methods, such as UV-vis absorption spectroscopy, fluorescence spectroscopy, fourier transform infrared spectroscopy (FT-IR), fluorescence lifetime, transmission electron microscope (TEM) and X-ray photoelectron spectroscopy (XPS), were applied for the analysis of trypsin-templated copper nanoclusters (Cu NCs@Tryp). The Cu NCs@Tryp released green fluorescence at maximum emission wavelength of 457 nm under maximum excitation wavelength of 377 nm. More importantly, the fluorescence of Cu NCs@Tryp was efficiently quenched by Bai. According to this phenomenon, a facile, rapid and selective turn-off fluorescence probe for Bai sensing was developed. Under the optimized testing conditions, the ln(F0/F) value and concentration of Bai displayed excellent linear relationship changing from 0.5 to 60 µM (R2 = 0.9969), and the detection limit was 0.078 µM. Furthermore, the Cu NCs@Tryp has been successfully employed to measure the amount of Bai in bovine serum samples with satisfactory recoveries.

circ­Grm1 promotes pulmonary artery smooth muscle cell proliferation and migration via suppression of GRM1 expression by FUS.

Pulmonary arterial hypertension is a progressive and fatal disease. Recent studies suggest that circular RNA (circRNAs/circs) can regulate various biological processes, including cell proliferation. Therefore, it is possible that circRNA may have important roles in pulmonary artery smooth muscle cell proliferation in hypoxic pulmonary hypertension (HPH). The aim of the present study was to determine the role and mechanism of circRNA­glutamate metabotropic receptor 1 (circ­Grm1; mmu_circ_0001907) in pulmonary artery smooth muscle cell (PASMC) proliferation and migration in HPH. High­throughput transcriptome sequencing was used to screen circRNAs and targeted genes involved in HPH. Cell Counting Kit­8 (CCK­8), 5­ethynyl­2­deoxyuridine and wound healing assays were employed to assess cell viability and migration. Reverse transcription­quantitative PCR and western blotting were used to detect target gene expression in different groups. Bioinformatical approaches were used to predict the interaction probabilities of circ­Grm1 and Grm1 with FUS RNA binding protein (FUS). The interactions of circ­Grm1, Grm1 and FUS were evaluated using RNA silencing and RNA immunoprecipitation assays. The results demonstrated that circ­Grm1 was upregulated in hypoxic PASMCs. Further experiments revealed that the knockdown of circ­Grm1 could suppress the proliferation and migration of hypoxic PASMCs. Transcriptome sequencing revealed that Grm1 could be the target gene of circ­Grm1. It was found that circ­Grm1 could competitively bind to FUS and consequently downregulate Grm1. Moreover, Grm1 could inhibit the function of circ­Grm1 by promoting the proliferative and migratory abilities of hypoxic PASMCs. The results also demonstrated that circ­Grm1 influenced the biological functions of PASMCs via the Rap1/ERK pathway by regulating Grm1. Overall, the current results suggested that circ­Grm1 was associated with HPH and promoted the proliferation and migration of PASMCs via suppression of Grm1 expression through FUS.

The Influence of Bosentan on MicroRNA-27a/PPARγ/ET-1 Signaling Pathway in Pulmonary Artery Hypertension.

Pulmonary artery hypertension (PAH) is a common and serious disease which is characterized by pulmonary vascular remodeling. Bosentan (BST) is the first approved oral targeted drug of endothelin-1 (ET-1) receptor antagonists for the treatment of PAH. MicroRNA-27a (miR-27a) and peroxisome proliferator-activated receptor γ (PPARγ) were found to be related to the pathogenesis of PAH. To further explore the signal transduction mechanism of BST in the treatment of PAH, we examined the effects of BST on endothelin receptors, miR-27a, and PPARγ. Meanwhile, the influence of miR-27a in the formation and development of PAH was discussed. Our results demonstrated that during the pathophysiology of PAH, miR-27a, PPARγ, and ET-1 were cross-inhibited, which indicated that the miR-27a/PPARγ/ET-1 signaling pathway was dysregulated; in addition, BST could competitively bind to ET-1 receptors and inhibit the miR-27a/PPARγ/ET-1 signaling pathway, thereby delaying the proliferation of PASMCs and affecting the development of PAH. Our results give a new understanding of the pathogenesis and treatment of PAH and provide more reliable evidence for the application of BST in the treatment of PAH in the clinic.

Highly sensitive and selective fluorescence sensing of nitrofurantoin based on water-soluble copper nanoclusters.

In this contribution, dopamine-protected copper nanoclusters as a novel fluorescent nanosensor was employed to detect nitrofurantoin (NFT) for the first time, which were prepared by using dopamine as the stabilizing agent and sodium borohydride (NaBH4) and hydrazine hydrate (N2H4·H2O) as the reducing agents. A series of methods were used to analyze the structure and optical properties of as-prepared Cu NCs, such as UV-Vis absorption spectroscopy, fluorescence spectroscopy, Fourier transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). It possessed high dispersion, excellent water solubility, good photostability and strong green fluorescence under UV lamp (365 nm). Significantly, the FL intensities of Cu NCs were quenched with addition of NFT. The analytical method possessed good linear relationship between the relative fluorescence intensity (F0/F) and the NFT concentrations (range from 5 to 120 µM), and the limit of detection (LOD) could reach 0.73 µM. The fluorescence detection mechanisms were attributed to the static quenching and inner filter effect (IFE). In addition, this proposed fluorescence sensor has been successfully used for the detection of NFT in bovine serum samples.

Anatomical Findings and Short-term Efficacy of Fascial Anatomy-guided Infrapyloric Lymphadenectomy in Laparoscopic Radical Gastrectomy for Gastric Cancer.

OBJECTIVE: The aim was to investigate the anatomical layers of the specific fascia involved in infrapyloric lymphadenectomy in laparoscopic radical gastrectomy for gastric cancer and to analyze the short-term efficacy of an anatomy-guided surgical approach. MATERIALS AND METHODS: On the basis of many years of clinical practice in fascial anatomy-guided laparoscopic radical gastrectomy for gastric cancer, we proposed anatomical considerations for infrapyloric lymphadenectomy in this procedure and investigated the anatomy of the mesentery and mesenteric fusion in this region, including the specific starting and ending points and the plane of the operation. We also retrospectively analyzed the clinical data of 265 patients who underwent fascial anatomy-guided infrapyloric lymphadenectomy in laparoscopic radical gastrectomy for gastric cancer from January 2015 to January 2019 and compared the short-term efficacy between the fascial anatomy-guided laparoscopic infrapyloric lymphadenectomy plus mesogastric excision group and the laparoscopic infrapyloric lymphadenectomy group. RESULTS: Extensive mesenteric fusion and folds exist in the infrapyloric region of the stomach, and removal of the medial fold (medial leg) and lateral fold (lateral leg) of the infrapyloric mesogastrium during surgery is easily missed, resulting in incomplete removal of the infrapyloric mesogastrium. Baseline data were comparable between the laparoscopic infrapyloric lymphadenectomy plus mesogastric excision group and the laparoscopic infrapyloric lymphadenectomy group. The mean operative time for infrapyloric lymphadenectomy, the number of positive lymph nodes harvested in the infrapyloric region, and the number of patients with mesenteric metastasis in the infrapyloric region were not significantly different (P>0.05). The number of harvested lymph nodes was higher in the laparoscopic infrapyloric lymphadenectomy plus mesogastric excision group than in the laparoscopic infrapyloric lymphadenectomy group (5.09±3.30 vs. 4.13±2.90, P<0.05), and intraoperative blood loss was lower in the former group than in the latter group (5.89±3.78 vs. 25.21±11.24 mL, P=0.000). CONCLUSION: Fascial anatomy-guided laparoscopic infrapyloric lymphadenectomy enables systematic and complete removal of the lymph nodes and mesentery of the infrapyloric region with less intraoperative blood loss.

Blue-emitting glutathione-capped copper nanoclusters as fluorescent probes for the highly specific biosensing of furazolidone.

Herein, a facile, straightforward and green method was developed to prepare copper nanoclusters by using glutathione (GSH) as the protecting agent and ascorbic acid as the reducing agent. The glutathione-templated copper nanoclusters (GSH-Cu NCs) were characterized through fluorescence spectroscopy, UV-vis absorption spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and fluorescence lifetime analysis. The as-synthesized Cu NCs showed blue fluorescence with a peak centered at 426 nm. The Cu NCs had excellent water solubility, stability and dispersibility. Based on the inner filter effect and static quenching mechanism, Cu NCs were employed to detect furazolidone in bovine serum samples. Under optimal detection conditions, a good linear relationship was observed between F0/F and the furazolidone concentration from 0.05 to 60 µM. The detection limit (LOD) was 0.012 µM. Furthermore, the fluorescence probe was successfully used in the quantification of furazolidone in bovine serum samples. In addition, this analytical method provides a rapid, easy and ultrasensitive fluorescence platform for the detection of furazolidone.

MiR-1/133 attenuates cardiomyocyte apoptosis and electrical remodeling in mice with viral myocarditis.

BACKGROUND: The role of miR-1 and miR-133 in regulating the expression of potassium and calcium ion channels, and mediating cardiomyocyte apoptosis in mice with viral myocarditis (VMC) is investigated herein. METHODS: Male Balb/c mice were randomly divided into groups: control group, VMC group, VMC + miR-1/133 mimics group, or VMC + miR-1/133 negative control (NC) group. VMC was induced with coxsackievirus B3 (CVB3). MiR-1/133 mimics ameliorated cardiac dysfunction in VMC mice and was compared to the VMC+NC group. RESULTS: Hematoxylin and eosin staining showed a well-arranged myocardium without inflammatory cell infiltration in the myocardial matrix of the control group. However, in the VMC and VMC+NC groups, the myocardium was disorganized and swollen with necrosis, and the myocardial matrix was infiltrated with inflammatory cells. These changes were alleviated by miR-1/133 mimics. TUNEL staining revealed decreased cardiomyocyte apoptosis in the VMC + miR-1/133 mimics group compared with the VMC group. In addition, miR-1/133 mimics up-regulated the expression of miR-1 and miR-133, the potassium channel genes Kcnd2 and Kcnj2, as well as Bcl-2, and down-regulated the expression of the potassium channel suppressor gene Irx5, L-type calcium channel subunit gene a1c (Cacna1c), Bax, and caspase-9 in the myocardium of VMC mice. MiR-1/133 also up-regulated the protein levels of Kv4.2 and Kir2.1, and down-regulated the expression of CaV1.2 in the myocardium of VMC mice. CONCLUSIONS: MiR-1 and miR-133 decreased cardiomyocyte apoptosis by mediating the expression of apoptosis-related genes in the hearts of VMC mice.

Ascorbic acid stabilised copper nanoclusters as fluorescent sensors for detection of quercetin.

In this report, green-emitting fluorescence copper nanoclusters (Cu NCs) were synthesized using ascorbic acid as reducing agent and protecting agent. The ascorbic acid capped Cu NCs (AA-Cu NCs) were characterized using fluorescence spectroscopy, UV-vis absorption spectroscopy, Fourier Transform Infrared Spectroscopy (FT-IR), Transmission Electron Microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). The analysis data demonstrated that the AA-Cu NCs were highly dispersed with an average diameter of 2 nm. The as-prepared Cu NCs possessed good water solubility, excellent photostability and displayed excitation-dependent fluorescence characteristics. More importantly, the fluorescence intensity of AA-Cu NCs was linearly quenched in the presence of quercetin from 0.7 to 50 µM and the detection limit (LOD) was 0.19 µM. Finally, the fluorescence sensor was successfully employed to detect quercetin in bovine serum samples.

New pathogenic variant of BMPR2 in pulmonary arterial hypertension.

OBJECTIVES: The aim of this study was to evaluate the variant frequency of pulmonary arterial hypertension-related genes and provide theoretical basis for genetic screening of patients with pulmonary arterial hypertension further. METHODS: Ten genes associated with pulmonary arterial hypertension were sequenced in 7 cases of idiopathic pulmonary arterial hypertension and 34 cases of congenital heart disease (CHD) associated with pulmonary arterial hypertension by next-generation high-throughput sequencing. Function prediction and gene variant amino acid conservation were carried out by bioinformatics software. Family study was performed on the patients with the variant. RESULTS: A new bone morphogenetic protein receptor type 2(BMPR2) variant (c.344T>C, p. F115S) was discovered in a girl who was diagnosed with idiopathic pulmonary arterial hypertension. Her second aunt and third aunt carried the same variant and were confirmed as patients with pulmonary arterial hypertension as well. No variants or single nucleotide polymorphisms were found in other pulmonary arterial hypertension-associated genes. CONCLUSIONS: BMPR2 variant is the most common variant of pulmonary arterial hypertension. Genetic screening of BMPR2 variant and family survey in patients with pulmonary arterial hypertension is suggested for the sake of definite cause and better treatment.

Vascular repair and anti-inflammatory effects of soluble epoxide hydrolase inhibitor.

Kawasaki disease (KD) is the leading cause of acquired heart disease in pediatric patients in developed countries. Coronary artery aneurysms and myocardial infarction may occur if the disease remains untreated. An estimated 10-20% of KD patients do not respond to intravenous gamma globulin (IVIG), and thus, alternative treatments are currently being investigated. Epoxyeicosatrienoic acids (EETs) are natural anti-inflammatory factors and angiogenic mediators degraded by soluble epoxide hydrolase (sEH). sEH inhibitory factors have been demonstrated to stabilize EET levels, inhibit inflammation and promote vascular repair in vivo. The present study aimed to determine whether an increase in EET levels induced by treatment with the sEH inhibitor 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA) promotes vascular repair in human coronary arterial endothelial cells (HCAECs) and reduces inflammation in a mouse model of KD induced by Lactobacillus casei cell wall extract. The effect of AUDA on vascular repair in HCAECs was assessed by using cell proliferation, migration, adhesion and tube formation assays, and the anti-inflammatory effect of AUDA in the mouse model of KD was determined by detecting the expression of matrix metalloproteinase (MMP)-9, tumor necrosis factor (TNF)-α and interleukin (IL)-1ß at the protein level via ELISA. The results demonstrated that AUDA increased the proliferation, migration, adhesion and tube formation ability of HCAECs in a dose-dependent manner. Furthermore, in the mouse model of KD, AUDA reduced the protein expression of MMP-9, IL-1ß and TNF-α, indicating that AUDA may alleviate inflammatory reactions in the coronary arteries of KD model mice. The present results also indicate that these effects may be exerted through the peroxisome proliferator activated receptor γ signaling pathway. Taken together, the present study supports the potential utility of AUDA in the treatment of KD.

Twenty-four-hour urine NE level as a predictor of the therapeutic response to metoprolol in children with recurrent vasovagal syncope.

BACKGROUND: Vasovagal syncope (VVS) is a heterogeneous disorder that creates challenges for treatment. Metoprolol is an important therapeutic option for children with VVS. AIMS: The study examined the predictive value of 24-h urine norepinephrine (NE) levels in the assessment of the therapeutic efficacy of metoprolol for recurrent VVS in children. METHODS: Thirty-eight children with recurrent VVS and 20 healthy children were enrolled in our study. Twenty-four-hour urine NE levels were measured by LC-MS-MS. VVS children were diagnosed by BHUTT and/or SNHUTT, and received metoprolol treatment for 3 months. Symptom scoring was utilized to evaluate the therapeutic effect. A ROC curve was used to investigate the predictive value of 24-h urine norepinephrine levels. RESULTS: There exists significant correlation between 24-h urine NE levels and supine systolic and diastolic blood pressures. The 24-h urine NE levels of responders (40.75 ± 12.86 µg/24 h) were higher than those of nonresponders (21.48 ± 6.49 µg/24 h), and there was a significant difference between the two groups (P < 0.001). A ROC curve of the predictive value of 24 h urine NE levels revealed that the area under the curve was 0.926. A cutoff value for 24-h urine NE level of 34.84 µg/24 h produced both high sensitivity (70%) and specificity (100%) in predicting the efficacy of metoprolol therapy for VVS. CONCLUSIONS: Patients with high 24-h urine NE levels have higher supine systolic and diastolic pressures and more effective responses to metoprolol. A 24-h urine norepinephrine level of > 34.84 µg/24 h was an indicator of the effectiveness of metoprolol therapy for VVS in children.

Association between the promoter methylation of the TBX20 gene and tetralogy of fallot.

OBJECTIVES: To investigate the association between promoter methylation of the TBX20 gene and tetralogy of Fallot (TOF)． Methods. The methylation level of TBX20 promoter regions in 23 patients with TOF and five controls were analyzed through bisulfite sequencing polymerase chain reaction. Meanwhile, the expression of TBX20 mRNA was measured using real time fluorescence quantitative polymerase chain reaction. RESULTS: The region -400 to -48 in the TBX20 promoter consisting of 42 CpG sites was predicted to contain multiple transcription factor binding sites. In this study, the overall methylation level in this region was lower in patients with TOF than in the controls (P = .035). Among the 42 CpG sites, the methylation percentages of the CpG 26 site in the TOF cases were lower than those in the controls (P = .016). The mRNA expression of TBX20 in the right ventricular outflow tract myocardium was increased in TOF cases in contrast to those in the controls (P < .001). The methylation levels in TOF cases were correlated with mRNA expression values (r = -0.81, P < .001). CONCLUSION: The downregulated methylation level at TBX20 promoter may be responsible for the elevated mRNA expression levels in patients with TOF. The abnormal methylation status of the TBX20 promoter may contribute to the pathogenesis of TOF.

Urotensin II contributes to collagen synthesis and up-regulates Egr-1 expression in cultured pulmonary arterial smooth muscle cells through the ERK1/2 pathway.

AIM: The objective of this study was to investigate the effects of urotensin II (UII) treatment on the proliferation and collagen synthesis of cultured rat pulmonary arterial smooth muscle cells (PASMCs) and to explore whether these effects are mediated by mitogen-activated protein kinase (MAPK) signaling pathways and early growth response 1 (Egr-1). METHODS: The proliferation of cultured PASMCs stimulated with different doses of UII was detected by BrdU incorporation. The mRNA expression levels of procollagen I (procol I), procollagen III (procol III), extracellular regulated protein kinase 1/2 (ERK1/2), stress-stimulated protein kinase (Sapk), p38 MAPK (p38), and Egr-1 mRNA in cultured PASMCs after treatment with UII, the UII-specific antagonist urantide, and the ERK1/2 inhibitor PD98059 were detected by real-time polymerase chain reaction (PCR), and the protein expression levels of procol I, procol III, phosphorylated (p)-ERK1/2, p-Sapk, p-p38, and Egr-1 were detected by Western blotting. RESULTS: Treatment with UII increased the proliferation of cultured PASMCs in a dose-dependent manner (P<0.05). However, treatment with urantide and PD98059 inhibited the promoting effect of UII on PASMC proliferation (P<0.05). Real-time PCR analysis showed that UII up-regulated the expression of procol I, procol III, ERK1/2, Sapk, and Egr-1 mRNA (P<0.05), but not p38 mRNA. However, the up-regulating effect of UII was inhibited by PD98059 and urantide. Western blotting analysis showed that UII increased the synthesis of collagen I, collagen III, p-ERK1/2, p-Sapk, and Egr-1, and these effects also were inhibited by PD98059 and urantide (P<0.05). CONCLUSIONS: Egr-1 participates in the UII-mediated proliferation and collagen synthesis of cultured rat PASMCs via activation of the ERK1/2 signaling pathway.