ABSTRACT
As a reversible and dynamic epigenetic marker, N6-adenylate methylation (m6A) modification is the most common mRNA modification in eukaryotes. This paper briefly described how m6A can influence RNA splicing, stability, and translation after transcription, and then participate in a variety of signaling pathways and biological and pathological processes, regulating cell proliferation, apoptosis, epithelial mesenchymal transformation (EMT) processes, and tumor invasion and metastasis. In addition, according to current studies, m6A methyltransferases (writers) are believed to promote EMT and tumor development, and readers and erasers both promote and inhibit EMT in different research objects. In this review, we summarized the mechanism of m6A modification and its role in cell transformation, and pointed out the direction of disease treatment.
ABSTRACT
Background Occupational and environmental particulate matter may cause fibrosis, accompanied by RNA m6A modification changes. Neodymium oxide (Nd2O3) can cause mouse lung fibrosis, which contains a large number of fibroblasts. Objective To investigate m6A modification of tumor necrosis factor receptor-associated protein 6/nuclear factor-κB (TRAF6/NF-κB) signaling pathway in fibrosis of human embryonic lung fibroblasts induced by Nd2O3, and identify the key m6A modification sites of TRAF6. Methods Designed concentrations of Nd2O3 (0, 1.563, 3.125, 6.25, 12.5, 25, 50, 100, and 200 mg∙L−1) were infected with HELF cells for 24 and 48 h, and cell viability was detected to determine exposure time and dose. Measurements included indicators of fibrosis [hydroxyproline (HYP) and transforming growth factor-β1 (TGF-β1)], m6A methylation level, methyltransferases (METTL3 and METTL14), demethylases (FTO and ALKBH5), reading proteins (YTHDC2 and YTHDF2), fibrosis-associated genes (collagen-І, vimentin, and α-SMA), and proteins related to signaling pathway (TRAF6, NFKB1, P65, and P-P65). The enrichment of m6A in TRAF6 mRNA was measured by methylated RNA immunoprecipitation-quantitative real-time PCR (MeRIP-qPCR). Results The results of cell viability indicated that 6.25, 12.5, 25 mg∙L−1 Nd2O3 and 48 h exposure time were used for subsequent experiments. After 48 h exposure, compared with the control group, the HYP level in the 25 mg∙L−1 Nd2O3 group was increased, and the levels of TGF-β1 in the 6.25, 12.5, and 25 mg∙L−1 Nd2O3 groups were increased (P<0.05); the overall m6A methylation levels of HELF cells in the 12.5 and 25 mg∙L−1 Nd2O3 groups were increased (P<0.05). At mRNA level, compared with the control group, the mRNA expression levels of methyltransferases METTL3 and METTL14 (6.25, 12.5, and 25 mg∙L−1 Nd2O3) were increased (P<0.05); the mRNA expression level of reading protein YTHDF2 (6.25, 12.5, and 25 mg∙L−1 Nd2O3) was increased (P<0.05), while the mRNA expression level of YTHDC2 (25 mg∙L−1 Nd2O3) was decreased (P<0.05); the mRNA expression levels of demethylases FTO (12.5 and 25 mg∙L−1 Nd2O3) and ALKBH5 (25 mg∙L−1 Nd2O3) were decreased (P<0.05); the mRNA expression levels of fibrosis-related genes vimentin, α-SMA, and collagen-Ⅰ (6.25, 12.5, and 25 mg∙L−1 Nd2O3) were increased (P<0.05); the mRNA expression levels of pathway-related genes TRAF6 (25 mg∙L−1 Nd2O3) and NFKB1 (12.5 and 25 mg∙L−1 Nd2O3) were increased (P<0.05). At protein level, compared with the control group, the expression levels of methyltransferases METTL3 (25 mg∙L−1 Nd2O3) and METTL14 (12.5 and 25 mg∙L−1 Nd2O3) were increased (P<0.05); the expression level of reading protein YTHDF2 (12.5 and 25 mg∙L−1 Nd2O3) was increased, while the expression level of YTHDC2 (25 mg∙L−1 Nd2O3) was decreased (P<0.05); the expression level of demethylase FTO (25 mg∙L−1 Nd2O3) was decreased (P<0.05); the expression level of fibrosis-associated protein vimentin was increased at 25 mg∙L−1 Nd2O3, and the expression levels of α-SMA and collagen-Ⅰ were increased at 12.5 and 25 mg∙L−1 Nd2O3 (P<0.05); the expression levels of TRAF6 and P-P65 were increased at 25 mg∙L−1 Nd2O3 (P<0.05). The MeRIP-qPCR results showed that compared with the control group, the concentrations of m6A in all Nd2O3 groups were significantly increased (P<0.05). Conclusions Upon exposure of HELF cells to Nd2O3, the alterations in fibrosis-related indexes increase the expression of some m6A methylases and decrease the expression of demethylases, thereby increasing the m6A methylase level, and may promote the progression of fibrosis by activating the TRAF6/NF-κB signaling pathway.
ABSTRACT
Background Arsenic can be toxic to human by triggering oxidative stress, which is companied by epigenetic modifications. Objective To investigate the modification of N6-methyladenosine (m6A) in human embryonic lung fibroblasts (HELF) during oxidative stress induced by sodium arsenite (NaAsO2). Methods HELF cells were treated by designed concentrations of NaAsO2 (0, 2.5, 5, 10, and 20 μmol·L−1) for 48 h. Cell viability was detected by 3-(4,5-dimethylthia zol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium (MTS) method; the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) as well as the content of malondialdehyde (MDA) were detected with corresponding kits; the level of m6A methylation in total RNA was detected by enzyme-linked immunosorbent assay; the mRNA expressions of m6A modified enzymes were detected by real-time fluorescence quantitative PCR, including methyltransferase-like 3 (METTL3), methyltransferase-like 14 (METTL14), Wilms' tumor 1-associated protein (WTAP), fat mass and obesity-associated protein (FTO), alkB family of Fe(II)/α-ketoglutarate-dependent dioxygenases 5 (ALKBH5), YTH domain containing protein 2 (YTHDC2), YTH domain family protein 2 (YTHDF2), and YTH domain family protein 3 (YTHDF3); the protein expressions of METTL3, FTO, YTHDC2, YTHDF3, and nuclear factor erythroid 2-related factor 2 (NRF2) were detected by Western blotting. The enrichment of m6A in NRF2 mRNA was detected by RNA methylated immunoprecipitation combined with real-time fluorescence quantitative PCR (MeRIP-qPCR). Results After the 0, 2.5, 5, 10, and 20 μmol·L−1 NaAsO2 treatment, the MTS results showed that compared with the control group, the cell viability of the 20 μmol·L−1 group decreased to 84% (P<0.05). The colorimetry results showed that compared with the control group, the activities of T-SOD in the 10 and 20 μmol·L−1 groups decreased (P<0.05); the activities of GSH-Px in the 2.5 and 10 μmol·L−1 groups decreased (P<0.05); the contents of MDA in the 10 and 20 μmol·L−1 groups increased. The results of enzyme-linked immunosorbent assay showed that the overall m6A methylation levels in the 0, 2.5, 5, 10, and 20 μmol·L−1 groups were (0.193 ± 0.023)%, (0.247 ± 0.021)%, (0.253 ± 0.006)%, (0.233 ± 0.006)%, and (0.262 ± 0.010)%, respectively, and compared with the control group, the m6A methylation levels in all the NaAsO2 treated groups increased (P<0.05). The real-time fluorescence quantitative PCR results showed that compared with the control group, the mRNA relative expression level of METTL3 decreased in the 2.5, 10, and 20 μmol·L−1 groups (P<0.05); the mRNA relative expression level of FTO decreased in the 20 μmol·L−1 group; the mRNA relative expression level of YTHDC2 increased in the 10 and 20 μmol·L−1 groups (P<0.05); the mRNA relative expression level of YTHDF3 increased in the 2.5, 10, and 20 μmol·L−1 groups (P<0.05). The Western blotting results showed that compared with the control group, the relative protein expression of METTL3 decreased in the 10 and 20 μmol·L−1 groups; the relative protein expression of FTO decreased in the 5 and 20 μmol·L−1 groups; the relative protein expression of YTHDC2 decreased in the 20 μmol·L−1 group (P<0.05); the relative nuclear protein expression of NRF2 decreased in the 10 and 20 μmol·L−1 groups (P<0.05). The MeRIP-qPCR results showed that m6A enrichment was significantly increased in the 20 μmol·L−1 NaAsO2 exposure group compared with the control group (P<0.05). After over-expression of FTO, the mRNA and protein relative expression levels of FTO and the relative expression level of nuclear protein of NRF2 in the FTO group were higher than those in the control group (P<0.05); the mRNA and protein relative expression levels of FTO in the NaAsO2 + FTO group and the nuclear protein expression level of NRF2 were higher than those in the NaAsO2 group (P<0.05). Conclusion In the process of oxidative stress induced by NaAsO2, m6A methylation level, m6A modified enzymes, m6A modification of NRF2 mRNA, and NRF2 expression could change in HELF cells.
ABSTRACT
Objective:To analyze the risk factors of lower extremities deep vein thrombosis after thoracoscopic surgery in elderly patients with lung cancer, establish a nomogram prediction model and conduct internal verification.Methods:A total of 183 elderly patients with lung cancer who underwent thoracoscopic radical resection in Nanchong Central Hospital from February 2018 to February 2022 were selected as the study subjects. According to the presence or absence of deep venous thrombosis of the lower extremities within one month after operation, the patients were divided into lower extremities deep venous thrombosis group ( n=61) and non-deep lower extremities venous thrombosis group ( n=122) . Univariate and multivariate analyses of deep venous thrombosis of lower extremities after thoracoscopic surgery for lung cancer in the elderly were performed, and a nomogram prediction model was constructed according to the multivariate analysis results, and the model was verified. Results:There were statistically significant differences in smoking history ( χ2=13.40, P<0.001) , preoperative chemotherapy ( χ2=8.79, P=0.003) , surgical method ( χ2=7.97, P=0.005) , operation time ( t=7.23, P<0.001) , postoperative bed rest time ( t=10.40, P<0.001) , combined with diabetes ( χ2=6.37, P=0.012) , combined with hyperlipidemia ( χ2=9.58, P=0.002) , preoperative D-dimer ( t=13.08, P<0.001) , preoperative fibrinogen ( t=5.84, P<0.001) and preoperative platelet count ( t=7.01, P<0.001) between the lower extremity deep venous thrombosis group and the non-lower extremity deep venous thrombosis group. The results of multivariate logistic regression analysis showed that preoperative chemotherapy ( OR=2.45, 95% CI: 1.05-5.71, P=0.038) , surgical method ( OR=2.55, 95% CI: 1.14-5.73, P=0.023) , postoperative bed rest time ( OR=1.50, 95% CI: 1.24-1.81, P<0.001) , combined with diabetes ( OR=3.60, 95% CI: 1.05-12.33, P=0.042) , and preoperative D-dimer ( OR=1.01, 95% CI: 1.01-1.01, P<0.001) were all independent risk factors for lower extremity deep vein thrombosis in elderly patients with lung cancer after thoracoscopic surgery. The C-index of nomogram for predicting lower extremity deep vein thrombosis-related factors was 0.86 (95% CI: 0.81-0.93) . The calibration curve showed that the model had a good correlation in predicting lower extremities deep venous thrombosis. Conclusion:Preoperative chemotherapy, surgical method, postoperative bed rest time, combined with diabetes, and postoperative D-dimer level are influence factors for lower extremity deep vein thrombosis in elderly patients with lung cancer after thoracoscopic surgery. The nomogram prediction model established in this study has high accuracy and discrimination for the prediction of lower extremity deep vein thrombosis in elderly patients with lung cancer after thoracoscopic surgery.
ABSTRACT
Objective To explore the curative effect and safety of uterine artery embolization combined with hysteroscope in treating cesarean scar pregnancy. Methods 36 cesarean scar pregnancy patients (CSP diameter under six cm by ultrasound) from June 2014 to June 2015 were selected and studied. All patients received the UAE and arterial injection of Methotrexate (MTX), and while the blood beta-HCG fell to around 1 000 u/L and the lesions thickness to serous membrane were over 0.2 cm by ultrasound, pregnancy lesions were resected by hysteroscopy surgery. Results The UAE and arterial injection of Methotrexate about (20.12 ± 3.85) min. Blood beta-HCG level and lesion diameter both fell or decreased, each with [(17 902.74±11 818.23) u/L vs (12 842.73 ± 8 525.73) u/L and (3.65 ± 1.02) cm vs (3.12 ± 0.97) cm] (t = 8.58, P < 0.01, t = 2.26, P < 0.05). Hysteroscopy surgery cost about (19.13 ± 2.67) min, the amount of bleeding were (17.43 ± 7.28) ml. Postoperative blood beta-HCG decreased to (113.45 ± 36.14) u/L within 2~4 days, the patients were discharged. Conclusion Hysteroscopy surgery with pretreatment of UAE combined with MTX injection could gain satisfactory outcome for patients with CSP. It has high success rate, low blood loss, fewer adverse reactions satisfactory outcome.
ABSTRACT
On the background of internationalization for higher education,we have reformed the educational model for bachelor students majoring in public health laboratory science and quarantine.The exploration and pilot study are focused on educational principles,curriculum planning,teaching patterns,experiments and practice,and teachem training.The four teaching strategies have been firstly recommended as follows:laying the strongest foundation of chemistry,expanding quarantine-related curricula for increasing quarantine capability,making full use of strength of clinical laboratory medicine,emphasizing English application.The curricula structure is refined and more social supporting resources have been got to make globalization to cultivate more health inspection and quarantine talents with multidisciplinary knowledge,abilities and strong adaptability.
ABSTRACT
We reported a simple and fast fluorescence system based on quantum dots ( QDs ) to detect glutamate dehydrogenase ( GLDH) , which inverted glutamate to α-ketogrutarate using NAD+ as a coenzyme. The fluorescence of CdTe QDs was quenched by nicotinamide adenine dimucleotide ( NAD+) through an electron transfer pathway, and the quencher NAD+ could be consumed by adding NAD+-dependent enzymes and corresponding substrates. Based on this principle we introduced GLDH to consume NAD+ in the QDs/NAD+ system, leading to the enhancement of the fluorescence intensity of QDs, which was in proportional to the amounts of GLDH added. Using this fluorescence system, we measured GLDH in a wide concentration range from 10 U/L to 1000 U/L, which was of significance in clinical diagnosis of different kinds of liver diseases.
ABSTRACT
OBJECTIVE:To study the therapeutic efficacy of atorvastatin for patients with carotid atherosclerotic plaques complicating cerebral infarction and the possible mechanism.METHODS:A total of 150 patients with cerebral infarction coexisting carotid atherosclerotic plaques were randomized to either control group (routine therapy against platelet aggregation) or trial group (routine therapy against platelet aggregation plus atorvastatin 20 mg?d-1).The scores of carotid atherosclerotic plaques,levels of blood lipid and C-reactive proteins (CRP) were compared between the two groups.RESULTS:At 6 and 12 months of follow-up,the scores of carotid atherosclerotic plaques in the trial group were significantly less than in the control group(P
ABSTRACT
OBJECTIVE To explore the risk factors and control strategies of nosocomial infection in clinical immunology laboratory.METHODS The positive detection rates of HBV marker and antibodies to HAV,HCV,HEV,HIV,TP,and TB in total of 91 877 samples in clinical immunology laboratory during Jul 2004 and Jul 2005 were retrospectively surveyed and statistically analyzed.RESULTS The number of positive specimens of HBV marker and antibodies to HAV,HCV,HEV,HIV,TP,and TB were respectively 8 376,7,26,24,107,3,522,and 52 and the positive detection rates of these items were respectively 16.68%,0.43%,0.20%,2.22%,6.49%,0.03%,4.75%,and 4.61%.CONCLUSIONS It is very important to understand the infection sources and risk factors in clinical immunology laboratory in order to strengthen management of hospital infection and protection of occupational exposure.
ABSTRACT
Objectives:To study the clinical characteristics of myopia in children and the measures to prevent it. Methods:748 patients with myopia were analyzed by refraction examination. Results:By the classification of the causes of myopia, the simple myopia constitutes the majority. By the degrees of the disease, the number of mild myopia was the first and moderate myopia was the second. By the patterns,the amount of compound myopic astigmatism was the majority. The correction of simple myopia or mild and moderate myopia was the most effective way. Of the patients(4～14 years), the age of 12 years had the highest myopia rate. Conclusions:To pay great attention to the prevention works of myopia in grade school years. Correcting refraction exactly and on time is an effective method to slow down the progression of myopia in children.
ABSTRACT
0 05). The serum levels of both IL-8 and TNF (P
ABSTRACT
Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.