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1.
Insects ; 14(1)2023 Jan 12.
Article in English | MEDLINE | ID: mdl-36662000

ABSTRACT

Plutella xylostella is a typical phototactic pest. LW-opsin contributes to the phototaxis of P. xylostella, but the expression changes of other genes in the phototransduction pathway caused by the mutation of LW-opsin remain unknown. In the study, the head transcriptomes of male G88 and LW-opsin mutants were compared. A GO-function annotation showed that DEGs mainly belonged to the categories of molecular functions, biological processes, and cell composition. Additionally, a KEGG-pathway analysis suggested that DEGs were significantly enriched in some classical pathways, such as the phototransduction-fly and vitamin digestion and absorption pathways. The mRNA expressions of genes in the phototransduction-fly pathway, such as Gq, ninaC, and rdgC were significantly up-regulated, and trp, trpl, inaD, cry1, ninaA and arr1 were significantly down-regulated. The expression trends of nine DEGs in the phototransduction pathway confirmed by a RT-qPCR were consistent with transcriptomic data. In addition, the influence of a cry1 mutation on the phototaxis of P. xylostella was examined, and the results showed that the male cry1 mutant exhibited higher phototactic rates to UV and blue lights than the male G88. Our results indicated that the LW-opsin mutation changed the expression of genes in the phototransduction pathway, and the mutation of cry1 enhanced the phototaxis of a P. xylostella male, providing a basis for further investigation on the phototransduction pathway in P. xylostella.

2.
Insect Sci ; 30(4): 1035-1045, 2023 Aug.
Article in English | MEDLINE | ID: mdl-36380712

ABSTRACT

Cryptochrome 1 (CRY1) functions as a light-responsive photoreceptor, which is crucial for circadian rhythms. The identity and function of CRY1 in Plutella xylostella remain unknown. In this study, cry1 was cloned and identified in P. xylostella. Then, a cry1-knockout strain (Cry1-KO) of P. xylostella with a 2-bp deletion was established from the strain Geneva 88 (G88) using the CRISPR/Cas9 technology. No daily temporal oscillation of cry1 was observed in G88 and Cry1-KO, and cry1 mean daily transcription of Cry1-KO was lower than that of G88. Both G88 and Cry1-KO demonstrated rhythmic locomotion under the light/dark condition with Cry1-KO being more active than G88 in the daytime, whereas Cry1-KO completely lost rhythmicity under constant darkness. The developmental period of pre-adult of Cry1-KO was longer than that of G88; the lifespan of the Cry1-KO male adult was shorter than that of G88; the fecundity of Cry1-KO was lower than that of G88; and Cry1-KO showed lower intrinsic rate of increase (r), net reproduction rate (R0 ), finite increase rate (λ), and longer mean generation time (T) than G88. Our results indicate that cry1 is involved in the regulation of locomotor circadian rhythm and development in P. xylostella, providing a potential target gene for controlling the pest and a basis for further investigation on circadian rhythms in lepidopterans.


Subject(s)
Circadian Rhythm , Cryptochromes , Male , Animals , Cryptochromes/genetics , Circadian Rhythm/genetics , Transcription Factors
3.
Front Psychol ; 13: 793483, 2022.
Article in English | MEDLINE | ID: mdl-35360560

ABSTRACT

Student-teacher relationships (STRs) have been examined by many studies. However, an omission still exists, the existing scales are not appropriate for studying STRs in private colleges because of the special character of these schools. This paper presents the development and validation of Private-College Student-Teacher Relationship Scale (PCSTRS), the first instrument to evaluate student-teacher relationships (STRs) in private colleges. The PCSTRS has six dimensions: trust, interaction, intimacy, care, approval, and comfort. In our main study, the validity and reliability of the six-factor PCSTRS model were demonstrated. The result of internal consistency coefficient indicated the high reliability of the scale, and the result of concurrent validity indicated the significant correlational relationships between the PCSTRS with other STR measures. In supplementary study, the PCSTRS was administered to 360 participants to confirm the applicability of PCSTRS and investigate the relation of STRs and students' traits, performance, and wellbeing, as well as the differences between the private school and the public school in this relation; the analyses revealed that there were significant differences in trust, intimacy, and care between private and public colleges; positive correlations were found between STRs and self-esteem, self-efficacy, academic performance, extracurricular activity involvement, and subjective wellbeing. Present research firstly develops the PCSTRS, examined the reliability and validity, and provides the proposed nomological network among related constructs.

4.
Front Plant Sci ; 13: 1002606, 2022.
Article in English | MEDLINE | ID: mdl-36605957

ABSTRACT

Huanglongbing (HLB), or citrus greening disease, has complex and variable symptoms, making its diagnosis almost entirely reliant on subjective experience, which results in a low diagnosis efficiency. To overcome this problem, we constructed and validated a deep learning (DL)-based method for detecting citrus HLB using YOLOv5l from digital images. Three models (Yolov5l-HLB1, Yolov5l-HLB2, and Yolov5l-HLB3) were developed using images of healthy and symptomatic citrus leaves acquired under a range of imaging conditions. The micro F1-scores of the Yolov5l-HLB2 model (85.19%) recognising five HLB symptoms (blotchy mottling, "red-nose" fruits, zinc-deficiency, vein yellowing, and uniform yellowing) in the images were higher than those of the other two models. The generalisation performance of Yolov5l-HLB2 was tested using test set images acquired under two photographic conditions (conditions B and C) that were different from that of the model training set condition (condition A). The results suggested that this model performed well at recognising the five HLB symptom images acquired under both conditions B and C, and yielded a micro F1-score of 84.64% and 85.84%, respectively. In addition, the detection performance of the Yolov5l-HLB2 model was better for experienced users than for inexperienced users. The PCR-positive rate of Candidatus Liberibacter asiaticus (CLas) detection (the causative pathogen for HLB) in the samples with five HLB symptoms as classified using the Yolov5l-HLB2 model was also compared with manual classification by experts. This indicated that the model can be employed as a preliminary screening tool before the collection of field samples for subsequent PCR testing. We also developed the 'HLBdetector' app using the Yolov5l-HLB2 model, which allows farmers to complete HLB detection in seconds with only a mobile phone terminal and without expert guidance. Overall, we successfully constructed a reliable automatic HLB identification model and developed the user-friendly 'HLBdetector' app, facilitating the prevention and timely control of HLB transmission in citrus orchards.

5.
Pest Manag Sci ; 77(7): 3519-3528, 2021 Jul.
Article in English | MEDLINE | ID: mdl-33837633

ABSTRACT

BACKGROUND: Opsins are crucial for animal vision. The identity and function of opsins in Plutella xylostella remain unknown. The aim of the research is to confirm which opsin gene(s) contribute to phototaxis of P. xylostella. RESULTS: LW-opsin, BL-opsin and UV-opsin, were identified in the P. xylostella genome. LW-opsin was more highly expressed than the other two opsin genes, and all three genes were specifically expressed in the head. Three P. xylostella strains, LW-13 with a 13-bp deletion in LW-opsin, BL + 2 with a 2-bp insertion in BL-opsin, and UV-29 with a 5-bp insertion and a 34-bp deletion in UV-opsin, were established from the strain G88 using the CRISPR/Cas9 system. Among the three opsin-knockout strains, only male and female LW-13 exhibited weaker phototaxis to lights of different wavelengths and white light than G88 at 2.5 lx due to defective locomotion, and LW-13 was defective to sense white, green and infrared lights. The locomotion of LW-13 was reduced compared with G88 at 2.5, 10, 20, 60, 80, 100, and 200 lx under the green light, but the locomotion of LW-13 female was recovered at 80, 100 and 200 lx. The defective phototaxis to the green light of male LW-13 was not affected by light intensity, while the defective phototaxis to the green light of female LW-13 was recovered at 10, 20, 60, 80, 100, and 200 lx. CONCLUSION: LW-opsin is involved in light sensing and locomotion of P. xylostella, providing a potential target gene for controlling the pest. © 2021 Society of Chemical Industry.


Subject(s)
Moths , Opsins , Phototaxis , Animals , CRISPR-Cas Systems , Clustered Regularly Interspaced Short Palindromic Repeats , Female , Gene Knockout Techniques , Male , Moths/genetics , Opsins/genetics
6.
Sci Rep ; 10(1): 11691, 2020 07 16.
Article in English | MEDLINE | ID: mdl-32678165

ABSTRACT

It has been shown that contrast-induced nephropathy (CIN) can be attenuated by the administration of PGE1. As an enzyme responsible for the production of PGE1, PTGS1 was confirmed in this study as a miR-512 target. Meanwhile, HULC has been identified as a competing endogenous RNA of miR-512. Therefore, in this study, we tested the diagnostic value of HULC and miR-512 in subjects with or without CIN. In addition, we evaluated the regulatory relationship among HULC, miR-512, PTGS1 and PGE1 in vitro. We enrolled 320 patients with coronary heart disease and divided them into a CIN group and a non-CIN group. Subsequently, we detected the differential expression of miR-512, HULC and PGE1 in the two groups. We also used a dual luciferase reporter assay to evaluate the regulatory relationship among HULC, miR-512, PTGS1 and PGE1 in THP-1 cells. In patients with CIN, the expression levels of HULC and PGE1 were lower, but the expression level of miR-512 was higher. MiR-512 could directly bind to and negatively regulate the expression of PTGS1 and HULC. The expression of HULC was positively correlated with the expression of PTGS1 and PGE1, while negatively correlated with the expression of miR-512. The findings of this study demonstrated that deregulation of lncRNA-HULC/miR-512/PTGS1/PGE1 might be involved in the pathogenesis of CIN.


Subject(s)
Alprostadil/metabolism , Contrast Media/adverse effects , Kidney Diseases/chemically induced , Kidney Diseases/metabolism , MicroRNAs/metabolism , RNA, Long Noncoding/metabolism , Signal Transduction/genetics , Aged , Apoptosis/genetics , Coronary Angiography/methods , Coronary Disease/surgery , Cyclooxygenase 1/genetics , Cyclooxygenase 1/metabolism , Female , Humans , Male , MicroRNAs/genetics , Middle Aged , Percutaneous Coronary Intervention/methods , RNA, Long Noncoding/genetics , THP-1 Cells , Transfection
7.
Arch Virol ; 164(5): 1335-1341, 2019 May.
Article in English | MEDLINE | ID: mdl-30848390

ABSTRACT

In infants, hepatitis B virus (HBV) infections are mainly acquired by mother-to-child transmission (MTCT). Current tests for the presence of HBV markers at birth can neither confirm nor exclude MTCT. The aim of this study was to find an early diagnostic marker of HBV MTCT. From 2011 to 2016, we studied a total of 5999 pregnant women who gave birth at our hospital in Shenzhen City, China. HBsAg-positive mothers and their offspring (n=386 pairs) were tested at birth for HBV markers, and 207 infants were followed up at 7-12 months after birth. The HBsAg-seropositive rate of the pregnant women was 12.5%. Additionally, 28.0%, 36.0%, 98.5% and 6.6% of umbilical cord (UC) blood samples of neonates were found to be positive for HBsAg, HBeAg, anti-HBc and HBV-DNA, respectively, whereas for neonatal femoral venous (FV) blood, the percentages were 16.2%, 38.0%, 98.8% and 2.6%, respectively. Mothers with high HBV DNA loads and those who were HBeAg positive were the most likely to have HBV-positive offspring. Immunoprophylaxis failed in five infants: the difference in median HBV DNA titer between UC blood from infants with and without HBV MTCT was statistically significant, and there was no significant difference in HBV DNA titer between UC blood and in peripheral blood of infants with HBV MTCT. In conclusion, we found that HBeAg positivity and high HBV loads are strong risk factors for MTCT of HBV and that the HBV DNA titer in the UC is a good predictor for HBV MTCT.


Subject(s)
Antibodies, Viral/blood , Hepatitis B Antibodies/blood , Hepatitis B e Antigens/blood , Hepatitis B virus/isolation & purification , Hepatitis B/diagnosis , Biomarkers , DNA, Viral/blood , Early Diagnosis , Female , Hepatitis B/epidemiology , Hepatitis B/transmission , Hepatitis B Core Antigens/immunology , Hepatitis B Surface Antigens/blood , Hepatitis B virus/genetics , Hepatitis B virus/immunology , Humans , Infant , Infant, Newborn , Infectious Disease Transmission, Vertical , Pregnancy , Pregnancy Complications, Infectious , Viral Load
8.
Nanoscale ; 8(13): 7155-62, 2016 Apr 07.
Article in English | MEDLINE | ID: mdl-26965185

ABSTRACT

This paper presents a novel method for enhancing the electroluminescence (EL) efficiency of ten-period silicon-rich oxide (SRO)/SiO2 superlattice-based light-emitting diodes (LEDs). A hydrogen ion beam (HIB) was used to irradiate each SRO layer of the superlattices to increase the interfacial roughness on the nanoscale and the density of the Si nanocrystals (Si NCs). Fowler-Nordheim (F-N) tunneling was the major mechanism for injecting the carriers into the Si NCs. The barrier height of the F-N tunneling was lowered by forming a nano-roughened interface and the nonradiative Pb centers were passivated through the HIB treatment. Additionally, the reflectance of the LEDs was lowered because of the nano-roughened interface. These factors considerably increased the slope efficiency of EL and the maximum output power of the LEDs. The lighting efficiency increased by an order of magnitude, and the turn-on voltage decreased considerably. This study established an efficient approach for obtaining bright Si NC/SiO2 superlattice-based LEDs.

9.
Heart Rhythm ; 12(10): 2132-40, 2015 Oct.
Article in English | MEDLINE | ID: mdl-25981147

ABSTRACT

BACKGROUND: Complete atrioventricular block (cAVB) has been deemed a rare complication after transcatheter closure for ventricular septal defect (VSD). However, this serious event appears to be underrecognized and is worth being investigated further. OBJECTIVES: To determine the incidence and predisposing factors of cAVB associated with closure of VSD using a modified double-disk occluder (MDO). METHODS: From December 21, 2001 to December 31, 2014, 1046 patients with perimembranous ventricular septal defect underwent percutaneous closure using the MDO. Electrocardiography was evaluated before the procedure, within 1 week after the procedure, and then at 1, 3, 6, and 12 months and every year thereafter. Other baseline and procedural parameters were also evaluated and a comparison between patients requiring pacemakers and those not suffering from cAVB was done. RESULTS: cAVB occurred in 17 patients (1.63%) after the procedure. Of the 17 patients, 8 underwent permanent pacemaker (PPM) implantation. The cAVB occurred within 30 days after the procedure in 14 patients and after 1 year in 3 patients. In comparison patients aged ≤18 years, patients aged >18 years were more prone to cAVB (P = .025). Logistic regression revealed no significant parameter to predict later requirement for PPM. CONCLUSIONS: The incidence of cAVB after transcatheter closure of VSD was acceptable, as part of the cAVB population recovered after administration of corticosteroid and application of a temporary pacemaker. Late cAVB (>1 year) appears to make it more difficult to restore normal conduction block. Because of the recurrence of cAVB, life-long follow-up with periodic electrocardiography examination may be mandatory.


Subject(s)
Atrioventricular Block/etiology , Cardiac Catheterization/methods , Cardiac Surgical Procedures/adverse effects , Heart Septal Defects, Ventricular/surgery , Septal Occluder Device/adverse effects , Adolescent , Adult , Aged , Atrioventricular Block/diagnosis , Atrioventricular Block/physiopathology , Cardiac Catheterization/adverse effects , Cardiac Surgical Procedures/instrumentation , Child , Child, Preschool , Echocardiography , Electrocardiography , Female , Follow-Up Studies , Humans , Infant , Male , Middle Aged , Prosthesis Failure , Retrospective Studies , Time Factors , Treatment Outcome , Young Adult
10.
Tumour Biol ; 34(1): 415-20, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23108892

ABSTRACT

In this study, we investigated whether COL4A3 mRNA expression levels were associated with clinical outcomes after treatment with a combination of gemcitabine (Gem)/cis-diamminedichloroplatinum(II) (CDDP) regimen for patients with advanced stage non-small cell lung cancer (NSCLC). Response and survival were correlated with the level of COL4A3 expression in 58 patients with advanced (stage IIIb or IV) NSCLC treated as part of a multicenter randomized trial with Gem 1,250 mg/m(2) on days 1 and 8 plus CDDP 100 mg/m(2) on day 1 every 3 weeks. mRNA was isolated from paraffin-embedded pretreatment primary tumor specimens, and relative expression levels of COL4A3/ß-actin were measured using a quantitative reverse transcription-PCR (Taqman) system. COL4A3 expression was detectable in all tumors. There were no significant differences in COL4A3 levels by gender, age, performance status, weight loss, or tumor stage. The overall response rate was 45.8 %. There were no significant associations between COL4A3 expression and response. Median overall survival was significantly longer in patients with low COL4A3 expression tumors compared to patients with high expression tumors. COL4A3 expression, Eastern Cooperative Oncology Group performance status, and presence of weight loss were significant prognostic factors for survival in a Cox proportional hazards multivariable analysis. These data suggest that COL4A3 expression is a predictive factor for survival after CDDP/Gem therapy in advanced NSCLC. Although there was a trend toward decreased response with high COL4A3 mRNA levels, this difference failed to reach statistical significance. This result may reflect the impact of Gem and the requirement for COL4A3 expression for CDDP/Gem synergism or may be attributable to the relatively small patient sample size in this study. Prospective studies of COL4A3 as a predictive marker for activity of CDDP-based regimens in NSCLC are warranted.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Autoantigens/metabolism , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/metabolism , Cisplatin/therapeutic use , Collagen Type IV/metabolism , Deoxycytidine/analogs & derivatives , Lung Neoplasms/drug therapy , Actins/analysis , Adult , Aged , Autoantigens/genetics , Biomarkers, Tumor/metabolism , Carcinoma, Non-Small-Cell Lung/mortality , Collagen Type IV/genetics , Deoxycytidine/therapeutic use , Female , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/mortality , Male , Middle Aged , Prognosis , RNA, Messenger/genetics , RNA, Messenger/metabolism , Treatment Outcome , Gemcitabine
11.
Zhonghua Xin Xue Guan Bing Za Zhi ; 40(4): 298-301, 2012 Apr.
Article in Chinese | MEDLINE | ID: mdl-22801307

ABSTRACT

OBJECTIVE: To evaluate the safety and efficacy of transcatheter closure of ruptured aneurysm of the sinus of Valsalva (RAVS) using a domestic made small-waist ventricular septal defect occluder. METHODS: Between September 2005 and December 2010, transcatheter closure of RAVS was randomly performed in 7 patients of (43.5 ± 13.0) years old using domestic made small-waist ventricular septal defect occluder in our department. RAVS was diagnosed by color Doppler echocardiography. The size of the small-waist double-disk occlude selected was 4 to 6 mm larger than the narrowest diameter of the opening of aneurysm. After establishment of the arteriovenous wire loop, the device was deployed by retrograde venous approach in all patients. All patients were followed up in terms of rhythm change, residual shunt, shape of occlude, and possible valve regurgitation by echocardiography for 6 to 12 months. RESULTS: All RAVS were confirmed by aortography. Aortography showed rupture of right coronary sinus into the right ventricle in 6 patients and non-coronary sinus ruptured into right ventricle in another patient. The estimated size of the defect was 4 to 7 (5.0 ± 1.3) mm by aortogram. Ventricular septal defect was evidenced in 2 patients. All defects were successfully occluded without complication. Two patients with ventricular septal defect received combined percutaneous ventricular septal defect closure. After transcatheter RAVS occlusion, pulmonary artery systolic pressure decreased from (35.6 ± 12.7) mm Hg (1 mm Hg = 0.133 kPa) to (27.4 ± 6.2) mm Hg (P < 0.05), and mean pulmonary artery pressure decreased from (21.9 ± 8.0) mm Hg to (16.1 ± 5.3) mm Hg (P < 0.05). Cardiac murmur disappeared right after successful occlusion. Echocardiography detected a trace of residual shunt in one patient after occlusion which disappeared the next day. There was no aortic regurgitation, hemolysis and arrhythmia during hospitalization. There was no device embolization, infective endocarditis, right heart failure and death during the 6 to 12 months follow-up. CONCLUSION: Transcatheter closure of RAVS with the domestic made small-waist ventricular septal defect occluder is safe and effective.


Subject(s)
Aneurysm, Ruptured/surgery , Cardiac Catheterization/instrumentation , Cardiac Catheterization/methods , Heart Aneurysm , Adult , Aged , Female , Humans , Male , Middle Aged , Septal Occluder Device , Sinus of Valsalva/pathology , Treatment Outcome , Young Adult
12.
Materials (Basel) ; 5(6): 1040-1054, 2012 Jun 07.
Article in English | MEDLINE | ID: mdl-28817023

ABSTRACT

The δ phase of plutonium with the fcc structure exhibits an unusual negative thermal expansion (NTE) over its narrow temperature range of stability, 593-736 K. An accurate description of the anomalous high-temperature volume effect of plutonium goes beyond the current capability of electronic-structure calculations. We propose an atomistic scheme to model the thermodynamic properties of δ-Pu based on the two-state model of Weiss for the Invar alloys, inspired by the simple free-energy analysis previously conducted by Lawson et al. The two-state mechanism is incorporated into the atomistic description of a many-body interacting system. Two modified embedded atom method potentials are employed to represent the binding energies of two competing electronic states in δ-Pu. We demonstrate how the NTE takes place in δ-Pu by means of Monte Carlo simulations implemented with the two-state mechanism.

14.
Case Rep Vasc Med ; 2011: 508171, 2011.
Article in English | MEDLINE | ID: mdl-22937464

ABSTRACT

A 19-year-old male patient presented cyanosis and dyspnoea because of the presence of multiple pulmonary arteriovenous fistulas resulting in oxygen desaturation. The CTA revealed that intestinal and splenic venous blood bypasses the liver and drains into the inferior vena cava. This is the first reported case of hepatopulmonary syndrome caused by congenital extrahepatic portosystemic shunt in which intestinal and splenic venous blood bypasses the liver and drains into the inferior vena cava.

15.
Zhonghua Xin Xue Guan Bing Za Zhi ; 38(4): 321-5, 2010 Apr.
Article in Chinese | MEDLINE | ID: mdl-20654076

ABSTRACT

OBJECTIVE: To evaluate the efficacy and adverse effects of transcatheter closure of perimembranous ventricular septal defect (pmVSD) with modified double-disk occluder device (MDVO). METHODS: Clinical data including clinical examination, electrocardiography daily after the procedure for a week, chest-X-rays and TTE before discharge and at 3-5 days after the procedure were analyzed from 604 patients underwent percutaneous closure of a pmVSD with MDVO at our department between December 2001 and December 2008. RESULTS: Procedure was successful in 576 out of 604 patients (95.4%) and 583 VSD occluders were placed. Endocarditis, thromboembolism, or deaths were not observed after procedure. Conduction block occurred in 81 patients (56 RBBB, 14 LBBB) and transient nonparoxysmal ventricular tachycardia in 31 patients after the procedure. Complete heart block occurred in 11 patients, 9 of them recovered in 3 weeks, permanent pacemaker was implanted in 2 patients (one had transient III degrees AVB before the procedure, the other underwent simultaneous closure of ventricular septal defect and atrial septal defect). Trivial/small residual shunts were found in 69 patients (12.0%). The residual shunts disappeared in 31 patients and remained unchanged in 38 patients (6.6%) 7 days after procedures. Aortic regurgitation developed in 5 patients (2 trivial/small, 3 small/moderate), and tricuspid regurgitation was present in 35 patients (32 trivial/small, 3 moderate). Five patients developed haemolysis (device retrieved via catheter in 1 patient due to persistent haemolysis, the other 4 patients recovered 3-14 days post procedure). Pseudoaneurysm of femoral artery occurred in 1 patient, and disappeared by pressure dressing. Device was successfully replaced in 2 patients with either device embolization (n = 1) or device misplacement (n = 1) after device retrieval by catheter. CONCLUSION: It is safe and effective to close congenital perimembranous ventricular septal defect with domestic-made occluder device.


Subject(s)
Cardiac Catheterization , Heart Septal Defects, Ventricular/therapy , Adolescent , Adult , Aged , Balloon Occlusion , Child , Child, Preschool , Echocardiography , Female , Humans , Infant , Male , Middle Aged , Treatment Outcome , Young Adult
16.
Circ Res ; 103(6): 591-7, 2008 Sep 12.
Article in English | MEDLINE | ID: mdl-18688046

ABSTRACT

Endothelium-derived nitric oxide (NO) is an important regulator of vascular function. NO is produced by endothelial NO synthase (eNOS), whose expression is downregulated by tumor necrosis factor (TNF)-alpha at the posttranscriptional level. To elucidate the molecular basis of TNF-alpha-mediated eNOS mRNA instability, eNOS 3' untranslated region (3'-UTR) binding proteins were purified by RNA affinity chromatography from cytosolic fractions of TNF-alpha-stimulated human umbilical vein endothelial cells (HUVECs). The formation of 3'-UTR ribonucleoprotein complexes, with molecular weight of 52 and 57 kDa, was increased by TNF-alpha. Matrix-assisted laser desorption ionization time-of-flight mass spectrometric analysis of the 52-kDa protein identified 3 peptides that comprise the peptide sequence of translation elongation factor 1-alpha 1 (eEF1A1). In HUVECs, TNF-alpha rapidly increased eEF1A1 expression, which is maximal after 1 hour and persists for up to 48 hours. RNA gel mobility-shift and UV cross-linking assays indicated that recombinant glutathione S-transferase-eEF1A1 fusion protein specifically binds to a UC-rich sequence in the 3'-UTR of eNOS mRNA. In addition, the domain III of eEF1A1 mediates the binding of eNOS 3'-UTR in eEF1A1. Overexpression of eEF1A1 markedly attenuated the expression of eNOS and luciferase gene fused with eNOS 3'-UTR in both COS-7 cells and bovine aortic endothelial cells (BAECs). Furthermore, adenovirus-mediated overexpression of eEF1A1 increased eNOS mRNA instability, whereas knockdown of eEF1A1 substantially attenuated TNF-alpha-induced destabilization of eNOS mRNA and downregulation of eNOS expression in HUVECs. These results indicate that eEF1A1 is a novel eNOS 3'-UTR binding protein that plays a critical role in mediating TNF-alpha-induced decrease in eNOS mRNA stability.


Subject(s)
Down-Regulation/physiology , Nitric Oxide Synthase Type III/antagonists & inhibitors , Nitric Oxide Synthase Type III/genetics , Peptide Elongation Factor 1/physiology , RNA Stability/genetics , Tumor Necrosis Factor-alpha/physiology , Amino Acid Sequence , Animals , COS Cells , Cattle , Cells, Cultured , Chlorocebus aethiops , Humans , Molecular Sequence Data , Nitric Oxide Synthase Type III/metabolism , Peptide Elongation Factor 1/genetics , Protein Biosynthesis/physiology
19.
J Clin Endocrinol Metab ; 92(5): 1875-80, 2007 May.
Article in English | MEDLINE | ID: mdl-17299069

ABSTRACT

CONTEXT: Obestatin, a sibling of ghrelin derived from preproghrelin, opposes ghrelin's effects on food intake. Plasma obestatin profiles in relation to ghrelin have not been fully investigated in human obesity. OBJECTIVE: We hypothesize that obesity might present with imbalance of circulating ghrelin and obestatin levels. PARTICIPANTS AND SETTING: Sixteen obese (eight men, aged 58.8 +/- 4.9 yr; eight women, aged 59.9 +/- 9.6 yr) and 14 normal-weight individuals (seven men, aged 52.7 +/- 5.9 yr; seven women, aged 56.1 +/- 4.9 yr) were evaluated at the in-patient department of Changhai Hospital, Shanghai, China. MAIN OUTCOME MEASURES: Total plasma ghrelin and obestatin levels, 1 h before and 2 h after breakfast, were measured by RIA. RESULTS: Both preprandial plasma ghrelin levels (P < 0.01) and obestatin levels (P < 0.01) were lower in the obese compared with normal-weight controls. However, unexpectedly, the ratio of preprandial ghrelin to obestatin was higher in obese compared with normal-weight controls (P < 0.01) even after adjustment for gender and age (P < 0.01). The ratio of postprandial ghrelin to obestatin was decreased both in obese (P < 0.05) and controls (P < 0.01) compared with their preprandial levels. There were no significant differences in the ratio of postprandial ghrelin to obestatin between obese and normal-weight controls. Body mass index was positively correlated with and was a significantly independent determinant of the preprandial ghrelin to obestatin ratio. CONCLUSION: Circulating preprandial ghrelin to obestatin ratio is elevated in human obesity. We suggest that high preprandial ghrelin to obestatin ratio may be involved in the etiology and pathophysiology of obesity.


Subject(s)
Eating/physiology , Obesity/blood , Peptide Hormones/blood , Blood Cell Count , Body Mass Index , Female , Ghrelin , Hormones/blood , Humans , Male , Middle Aged , Postprandial Period/physiology
20.
Zhonghua Yi Xue Za Zhi ; 86(38): 2698-702, 2006 Oct 17.
Article in Chinese | MEDLINE | ID: mdl-17199981

ABSTRACT

OBJECTIVE: To study the effects of fusion gene encoding the hVEGF(165) and fused hirudin on restenosis of injured carotid artery. METHODS: A fusion gene encoding hVEGF(165) and fused hirudin (hVEGF(165)-FH) was constructed and clone into the eukaryotic expression vector pcDNA3.0, thus constructing the plasmid VEGF(165)-FH/pcDNA3.0. Its activities to stimulate endothelial cell proliferation and to inhibit thrombosis were identified. Sixteen New Zealand rabbits underwent ligation of external carotid artery and a balloon was inserted into the common carotid artery for 30 minutes so as to construct model of restenosis of injured carotid artery. Then the rabbits were randomly divided into 4 equal groups. In the one-week and 3-week control groups, 400 microg of DNA of the plasmid pcDNA3.0 were transfused into the arterial lumen at the injured part immediately after the angioplasty, and 400 microg of DNA of the plasmid VEGF(165)-FH/pcDNA3.0 were transfused in the 1-week and 3-week experimental groups. One week and 3 weeks after the treatment peripheral blood samples were collected to detect the activated partial thromboplastin time (APTT), thrombin time (TT), and platelet aggregation rate, and then the rabbits underwent angiography to observe the situation of restenosis. Then the rabbits were killed to take out the injured part of artery to undergo pathological examination and Western blotting. RESULTS: The values of APTT, TT, and platelet aggregation rate were not significantly different among the 4 groups. Angiography conducted 1 and 3 weeks later showed that restenosis was significantly mild in the 2 experimental groups in comparison with the 2 control groups, and severe restenosis was seen in the 3-week control group. Western blotting showed that expression of specific fused protein could be found in the 1-week and 3-week experimental group, the amount of the latter group being less than that of the former group; however, no expression of specific fused protein was found in the 2 control groups. Pathological examination showed that the narrowing of lumen 1-week and 3-week experimental groups were 11.50% and 19.75%, both significantly milder than those of the 1-week and 3-week control groups (33.25% and 52.25% respectively, both f P < 0.05). VB staining showed that the (intima/media (I/M) ratio of the 1-week and 3-week experimental groups were 0.12 and 0.35 respectively, both significantly lower than those of the 2 control groups (0.50 and 1.07 respectively, both P < 0.05). CONCLUSION: Accelerating re-endothelialization and inhibiting thrombosis, the fused gene hVEGF(165)-FH effectively prevents restenosis after angioplasty, On the basis of endothelial repair, construction of fused genes with double even multiple targets may be a novel and potential therapeutic approach for restenosis after percutaneous coronary intervention.


Subject(s)
Carotid Stenosis/therapy , Genetic Therapy/methods , Hirudins/genetics , Vascular Endothelial Growth Factor A/genetics , Animals , Blotting, Western , Carotid Artery Injuries/complications , Carotid Stenosis/etiology , Hirudins/metabolism , Partial Thromboplastin Time , Rabbits , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Recombinant Fusion Proteins/physiology , Thrombin Time , Time Factors , Vascular Endothelial Growth Factor A/metabolism
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