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Objective@#To investigate the effects of electrochemically dealloying of Ti6Al4V abutments on human gingival fibroblasts (HGFs) and to provide experimental evidence for surface modification of implant abutments.@*Methods@#The samples were divided into an NC group (negative control, no other treatment on a smooth surface), an NM-1 group (nanomesh-1, electrochemical dealloying treatment in 1 mol/L NaOH 1 h on 2 V voltage), and an NM-2 group (nanomesh-2, electrochemical dealloying treatment in 5 mol/L NaOH 1 h on 2 V voltage). The surface morphologies of the samples and the adhesion of HGFs on the sample surfaces were observed with scanning electron microscopy (SEM). The surface hydrophilicities of the samples were measured with a contact angle measuring instrument. The proliferation of HGFs on the different samples were evaluated with CCK-8, and the expression of adhesion-related genes, including collagen Ⅰ (COL1A1), collagen Ⅲ (COL3A1), fibronectin 1 (FN1), focal adhesion kinase (FAK), vinculin (VCL), integrin α2 (ITGA2), and integrin β1 (ITGB1), on the different samples was measured with qRT-PCR. The expression of vinculin on the surfaces of HGFs was observed via confocal laser scanning microscopy (CLSM) after immunofluorescent staining. Collagen fiber secretion and syntheses of HGFs from different samples were evaluated via Sirius red staining.@*Results@#SEM revealed the formation of ordered and uniform three-dimensional mesh structures on the surfaces of the NM-1 and NM-2 groups, with grid diameters of approximately 30 nm for the NM-1 group and approximately 150 nm for the NM-2 group. Compared with that of the NC group, the water contact angles of the NM-1 group and NM-2 groups were significantly lower (P<0.000 1). Cell proliferation in the NM-1 group was significantly greater than that in the NC group (P<0.01). Moreover, there was no significant difference in the water contact angles or cell proliferation between the NM-1 group and the NM-2 group. SEM revealed that HGFs were adhered well to the surfaces of all samples, while the HGFs in the NM-1 and NM-2 groups showed more extended areas, longer morphologies, and more developed pseudopodia than did those in the NC group after 24 h. qRT-PCR revealed that the expression levels of the adhesion-related genes COL1A1, COL3A1, FN1, FAK and VCL in the NM-1 group were significantly greater than those in the NC and NM-2 groups (P<0.01). The expression of vinculin protein in the NM-1 group was the highest, and the number of focal adhesions was greatest in the NM-1 group (P<0.01). The results of Sirius red staining showed that the NM-1 group had the highest secretion and syntheses of collagen fibers (P<0.000 1).@*Conclusion@#The three-dimensional nanomechanical structure of Ti6Al4V modified by electrochemical dealloying promoted the adhesion, proliferation, collagen fiber secretion and syntheses of HGFs, and electrochemical dealloying of Ti6Al4V with a grid diameter of approximately 30 nm obviously promoted HGF formation.
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Octapeptin has strong antibacterial activity against Gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, while it also has activity against some Gram-positive bacteria. This study used natural octapeptin A3 and B3 as lead compounds for structural modification. Twenty-one peptide derivatives (including A3 and B3) containing eight amino acid residues were prepared by solid-phase synthesis, and evaluated for antibacterial activity and renal cytotoxicity. Among them, three compounds 6, 7 and 17 exhibited broad-spectrum antibacterial activity and significantly enhanced the activity for Gram-positive bacteria while maintaining the activity of Gram-negative bacteria. Several compounds improved the activity for Pseudomonas aeruginosa. Compound 7 was active against all test strains and had relatively low renal cytotoxicity. The results provide a basis for the further development of novel polypeptide antibiotics.
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Oral administration is the most convenient way of drug delivery， but due to the existence of intestinal barrier， the oral bioavailability of drugs is generally low， especially for drugs with low water solubility， poor permeability and macromolecules. For decades， researchers have demonstrated that nano-delivery system is one of the most effective strategies to solve this problem， but nano-delivery systems have shown limited improvement in the oral bioavailability of drugs. Therefore， researchers have proposed to use transporter-mediated nano-delivery systems to promote the oral absorption of drugs. The intestinal tract were highly expressed as a transporter for ingesting various nutrients（such as glucose， oligopeptides and bile acids）， which was an excellent target of oral drug delivery system. Its substrate were modified on the nano-delivery system， and the loaded drugs could cross the intestinal barrier and enter the systemic circulation more efficiently through the targeting effect of transporters. At present， more and more evidences supported the potential of transporters in the field of oral drug delivery system. Therefore， this paper reviewed the research on intestinal transporters-mediated nano-delivery system to promote oral absorption of drugs， including the distribution of intestinal transporters， three strategies of transporter substrate modification， the transport properties of different types of transporters and their effects of mediating the nano-delivery system for promoting the oral absorption of drugs or treating diseases， with the aim of providing an important theoretical reference for the development of intestinal targeted nano-delivery systems.
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RESUMEN Introducción: la anatomía arterial es la principal limitante para el abordaje aórtico endovascular estándar. Presentamos nuestra experiencia para la reparación endovascular de aneurismas aórticos complejos. Material y métodos: estudio observacional retrospectivo en pacientes con aneurismas complejos (yuxta/pararrenales y toracoabdominales) tratados en forma consecutiva mediante: endoprótesis fenestradas (FEVAR), ramificadas (BEVAR), con EndoAnchors (ESAR), o en chimenea (ChEVAR). La decisión de la técnica fue determinada con base en la anatomía arterial. Resultados: se evaluaron los últimos 50 procedimientos (6 mujeres; edad promedio 71,3 años; diámetro 69,6mm; 3 pacientes con aneurismas complicados), de los cuales 22 recibieron FEVAR (2,8 fenestraciones / paciente), 11 BEVAR, 11 ESAR y 6 ChEVAR (1,8 chimeneas /paciente). La tasa de éxito técnico fue del 100% (ausencia de endoleak I o III con permeabilidad adecuada de los vasos viscerales). A 30 días 3 pacientes fallecieron (6%). Durante el seguimiento, 5 pacientes presentaron oclusión de la arteria renal, repermeabilizada en 4. Cuatro pacientes desarrollaron un endoleak tipo IA (3 ESAR secundarios y un ChEVAR), un paciente un endoleak IC y un cuarto uno IIIB (22%, 3 de los 11 ESAR, ninguno de los FEVAR industriales). En el análisis de supervivencia, la supervivencia global fue del 88,6% al año, y libre de reoperación del 86,5%. Conclusiones: se trata de la primera publicación en nuestro medio que muestra un enfoque global del paciente con un aneurisma de aorta complejo, de acuerdo con sus características anatómicas. Estas tecnologías ya desempeñan un papel primario en el tratamiento de estos pacientes.
ABSTRACT Background: Arterial anatomy is the main limiting factor for standard endovascular aortic (EVAR) approach. We present our experience for endovascular repair of complex aortic aneurysms. Material and Methods: This is a retrospective observational study in patients with complex aneurysms (juxta/pararenal and thoracoabdominal) treated consecutively with: fenestrated (FEVAR), branched (BEVAR), EndoAnchors (ESAR), or chimney (ChEVAR) stents. The decision of the technique was determined based on the arterial anatomy. Results: The last 50 procedures were evaluated (6 women; mean age 71.3 years; diameter 69.6 mm; and 3 patients with complicated aneurysms), among whom 22 received FEVAR (2.8 fenestrated stents/patient), 11 BEVAR, 11 ESAR and 6 ChEVAR (1.8 chimney stents/patient). Technical success rate was 100% (absence of type I or III endoleak with adequate patency of the visceral vessels). Three patients died within the first 30 days (6%). During follow-up, 5 patients presented with renal artery occlusion, treated successfully in 4 cases. Four patients developed type IA endoleak (3 secondary ESAR and one ChEVAR), one patient IC endoleak and almost a quarter of cases type IIIB endoleak (22%, 3 out of 11 patients receiving ESAR, none of the industrial FEVAR group). In survival analysis, overall survival analysis was 88.6% at one year, and 86.5% of cases were free from reoperation. Conclusions: This is the first publication in our setting that shows a global approach to the patient with complex aortic aneurysm, according to the anatomical characteristics. These technologies already play a primary role in the treatment of these patients.
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Aim: This study sought to investigate the impact of training programs on hypertensive patients' awareness and practice of lifestyle modifications in out-patient clinics of the University College Hospital (UCH) Ibadan, Nigeria. Methodology: The study design was a cross-sectional survey conducted at the out-patient clinics of the University College Hospital (UCH), Ibadan, Nigeria. This setting was chosen due to the accessibility of hypertensive patients receiving regular care and follow-up. The sample size comprised 274 hypertensive patients attending the out-patient clinics for regular hypertension management. A simple random sampling method was employed to select study participants, ensuring a fair representation of the target population. A structured questionnaire was used to collect data from the participants and results were analysed using SPSS. Results: Findings indicate that the training programs greatly increased awareness of hypertension and lifestyle factors affecting hypertension for most participants (74.42%), with all participants recommending the programs. A significant majority of participants (95.74%) reported being on hypertension medication, while nearly all (99.22%) had received some form of training or counselling on lifestyle modification for hypertension management. Notably, all participants reported having made lifestyle changes following the training programs, with the majority rating the effectiveness of the programs as highly effective (57.36%). Further analysis indicated a statistically significant relationship between the effectiveness of the training programs and the participants' age, sex, educational level, and occupation. The most frequent lifestyle modification post-training was dietary changes, with 54.65% of participants reporting that they always engage in a healthy diet for hypertension management. However, the frequency of engagement in other lifestyle modifications, such as regular physical activity and weight management, was not as high. The majority of participants reported significant improvements in hypertension symptoms following the lifestyle modifications. Conclusion: These findings demonstrate that targeted training programs can significantly enhance hypertensive patients' awareness and practice of beneficial lifestyle modifications, thus improving health outcomes. Further, the study underscores the need for such training programs to be tailored according to the age, sex, educational level, and occupation of the patients to maximize their impact.
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Background: Obesity is described as a build-up of abnormal or excessive fat that risks life. A body mass index (BMI) is a screening tool for overweight and obesity. For a long time, BMI has been used as a marker to measure obesity. However, a significant limitation of using BMI is its failure to differentiate between a high body fat content and preserved or increased lean mass, especially in patients or subjects with a BMI <30 kg/m2. Normal weight obese (NWO) is such individuals who have high fat content but a normal BMI. To identify such individuals and to save them from morbidity associated with obesity, this study was carried out. Aims and Objectives: The objective of the study was to estimate the prevalence of NWO in healthy young adults (aged between 18 and 30 years) and to suggest the methods of primary prevention for obesity. Materials and Methods: In this observational cross-sectional study, the 324 subjects were selected randomly from the residents, students, and employees of Swami Vivekananda Subharti University. Subjects were chosen according to the study’s inclusion and exclusion criteria and also, based on the self-structure questionnaire. A general and systemic examination was carried out. The body composition of these subjects was done by bioelectric impedance analyzer BODY STAT QUAD SCAN 4000. Data were analysis using R-software (1.25 version), and unpaired t-test was applied. P < 0.05 was taken as significant in this study. Results: The prevalence of NWO in males was found to be 24.6% and females to be 13.4%. The body composition parameters of NWO and NWNO differed significantly. Conclusion: The increase rate of NWO is reason for concern and routine screening of body composition parameters should be done at health setups to identify that these NWO individuals and timely interventions can be made.
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Resumen En los últimos años, Colombia registra un aumento tanto de sobrepeso como de obesidad, aspecto que produce el establecimiento de modelos complejos de intervención que generen cambios de actitud, compromiso en las personas y el mejoramiento de hábitos alimenticios y de actividad física. Este estudio pretendió comparar la efectividad entre dos componentes psicológicos (estrategias conductuales tradicionales y un entrenamiento en aceptación y compromiso), sumados al entrenamiento físico concurrente y la educación nutricional (junto con una dieta hipocalórica), sobre variables antropométricas, la aptitud física, los aspectos de la conducta alimentaria y la sintomatología de salud mental. El diseño de investigación fue de caso único ABA (línea base-intervención-línea base) para cada condición de tratamiento. La muestra estuvo conformada por cinco mujeres y un hombre, entre los 33 y 49 años de edad, con preobesidad y obesidad tipo I, asignados entre los grupos de coaching conductual y entrenamiento en aceptación y compromiso. Los hallazgos muestran un mejoramiento en: composición corporal, resistencia aeróbica, fuerza prensil, compromiso y fusión cognitiva frente a situación de alimentación y actividad física en dos participantes de la estrategia de aceptación y compromiso, mientras que hubo mayor variación en los resultados de los participantes de coaching conductual. Este es un pilotaje de un estudio con mayor capacidad de inferencia.
Abstract There is an increase in both overweight and obesity in the world, aspect that tends for the approach of integral models of intervention, that generate changes in attitude, and commitment in people towards their eating habits and physical activity, that counteract a sedentary lifestyle and the influence of counterproductive processed foods for energy balance. This study aimed to compare the effectiveness between two psychological components (traditional behavioral strategies and a 6-session acceptance and commitment training), added together, each one: a) concurrent physical training (strength/endurance) with bioadaptation process, b) hypocaloric diet and c) Nutritional education. Measurements of anthropometric variables were obtained, physical fitness and mental health symptoms, while the average of steps was measured during the whole process, constituting this part in a single ABA case design for each treatment condition. The sample consisted of five women and one man, between 33 and 49 years of age, with pre-obesity and type I obesity, assigned in a non-random way between the behavioral coaching and acceptance and commitment training groups. In physical training, priority was initially given to the conditional capacity of strength, being an incident in the mobilization of large muscle groups (multi-joint) in order to generate a greater caloric expenditure, directly influencing exercises that optimize postural hygiene by working in the core zone and complementing training with synergistic or accompanying biarticular exercises in the movement of daily physical activities, subsequently the execution of aerobic resistance in search of the increase in the maximum oxygen consumption capacity ( max), ending with the implementation of vital flexibility exercises for physical exercise releasing muscle tension, favoring conditional physical ability. Regarding the intervention of the nutritional component, the most important input to begin to identify eating habits and subsequently modify them was the 24-hour reminder, applying in the first week of the baseline and the reversal phase. The nutritional intervention consisted of: (a) education so that participants understood and generated greater adherence to the nutritional proposal and (b) hypocaloric diet, reducing daily intake between 500 and 600 kilocalories per day. A follow-up anthropometric assessment was also carried out towards the third week of the intervention phase to obtain feedback on the incidence of their behavior on the anthropometric variables. Regarding the record of the number of steps, both in the baseline and reversion phases the monitor screens were covered, contrary to the intervention phase, since its observation allows self-monitoring towards the weekly personal goal of steps that was established and accompanied by a professional, who in turn generated a graph with the results of the week and gave feedback to each participant. The findings show an improvement in: body composition, aerobic endurance (VO2 max) and grip strength, being the main factor of result in the realization of the physical exercise. Likewise, an increase in the number of steps from the baseline is identified in both groups (X = 9 228.07; S = 1 985.89) to the intervention phase (X = 9 734.22; S = 1 660.86) and a decrease in reversion (X = 9 099.88; S = 2 328.69). In the acceptance and commitment group, no increase in mental health symptoms was identified, increased or sustained cognitive restriction and decreased scores for uncontrolled eating and emotional eating, while there was greater variation in the results of the behavioral coaching participants. This is a pilot of a study with greater capacity for inference.
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Treating obesity in children and adolescents is not a cakewalk. Asking them directly to get up and run, or just serving them a four-quadrant plate won’t serve the purpose at all. An obese child is already under the repercussions of oppressive remarks by the society. If at home as well, their mental milieu is not taken care of, there are chances of them being trapped in the shackles of depression. To avoid such drastic consequences, one must be prudent enough, while conversing with their children/patients regarding their weight. Motivation enhancement is very crucial in childhood obesity management, as the required changes in diet, behavior and physical activity are difficult to carry out with an insufficient level of motivation.1 The motivational hacks presented here might be helpful for parents and health care professionals in convincing their children/patients to stride through the road to health.
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Amplamente utilizados para substituir dentes perdidos, os implantes dentários nos últimos anos vêm apresentando tecnologias como superfícies com micro e nanotopografia e ajustes nas composições químicas, dentre outros, para melhorar a osseointegração e reduzir o prazo de tratamento, permitindo, assim, carga funcional imediata ou precoce em pacientes com densidade óssea reduzida. Vários métodos são aplicados com intuito de modificar a superfície do implante, como jateamento com areia, corrosão ácida, oxidação anódica, tratamento com flúor, usinagem, pulverização de plasma de titânio e revestimento de fosfato de cálcio; esses métodos podem aumentar notavelmente a área de superfície quando operada a técnica adequadada de modificação, quer por procedimento de adição ou subtração. Tais modificações promovem superfícies rugosas, as quais aumentam a porcentagem de contato osso-implante (BIC) durante o processo de cicatrização óssea inicial. Os principais benefícios da modificação da superfície são melhorar a molhabilidade (hidrofilicidade), adesão e fixação de células a implantes e proliferação celular. Dentre os tratamentos de superfície de implantes dentários destaca-se o jateamento de areia com granulação grossa e ataque-ácido com HCL/H2SO4 (SLA) em altas temperaturas, o revestimento de superfície do implante com hidroxiapatita, oxidação anódica e o duplo ataque ácido. O objetivo deste trabalho é realizar uma revisão de literatura discutindo a importância e eficácia desses métodos para a osseointegração e, por conseguinte, para a redução do período de tratamento.
Widely used to replace lost teeth, dental implants have been presenting technologies such as surfaces with micro and nano topography and adjustments in chemical compositions, among others, to improve osseointegration and reduce treatment time, thus allowing immediate or early functional load in patients with reduced bone density. Several methods are applied to modify the implant surface, such as sandblasting, acid corrosion, anodic oxidation, fluoride treatment, machining, titanium plasma spraying, and calcium phosphate coating; these methods can notably increase the surface area when the appropriate modification technique is operated, either by the addition or subtraction procedure. Such modifications promote rough surfaces, which increase the percentage of bone-implant contact (BIC) during the initial bone healing process. The main benefits of surface modification are to improve wettability (hydrophilicity), adhesion and attachment of cells to implants, and cell proliferation. Among the surface treatments for dental implants, sandblasting with large grit and acid-etching with HCL/H2SO4 (SLA) stands out at high temperatures. The surface coating of the implant with hydroxyapatite, anodic oxidation, and double acid-etching. This work aims to conduct a literature review discussing the importance and effectiveness of these methods for osseointegration and, therefore, for reducing the treatment period.
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Surface Properties , Therapeutics , Dental Implants , OsseointegrationABSTRACT
The occurrence of gastric cancer is closely related to environmental, genetic, and epigenetic factors. Currently, RNA modification is a research frontier and hotspot in the field of epigenetics. With the advancements in analytical chemistry and high-throughput sequencing technologies, new technologies and methods of exploring RNA modification are constantly being presented. Numerous studies have confirmed the involvement of RNA modifications in the occurrence and development of various diseases. Recent studies have shown that RNA modifications such as m6A, m5C, and ac4C regulate the malignant progression of various tumors, including gastric cancer, liver cancer, colorectal cancer, and leukemia. This article systematically reviews the research status and mechanism of different RNA modifications in the occurrence and development of gastric cancer, as well as discusses its potential value in the diagnosis and treatment of gastric cancer.
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N 6-methyladenosine (m 6A) is the most abundant epigenetic modification in eukaryotic messenger RNA (mRNA), which could be catalyzed by m 6A methyltransferase (Writers), recognized by methylation recognition enzymes (Readers), and removed by demethylase (Erasers). RNA splicing, translation, and stability could be modulated by m 6A methylation modification. The m 6A methylation modification is involved in the biological regulation of a variety of important functional genes in cellular activities. Importantly, abnormal m 6A modification affects the occurrence, development, metastasis and recurrence of tumors. Ionizing radiation can affect the level of m 6A and m 6A methylation-related enzymes. Recently, m 6A methylation is reported to regulate the efficacy of tumor radiotherapy by affecting DNA damage and radiosensitivity of tumor cells. In addition, ionizing radiation can also affect the level of m 6A modification in normal cells to regulate the progress of radiation-induced injuries. This review summarizes the research progress on the roles of m 6A methylation in tumor radiosensitivity and radiation-induced injuries, with the aim of providing novel strategies for the development of clinical tumor radiosensitizers and radioprotective agents.
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A variety of full 2ʹ-F/OMe-modified siRNAs were designed and synthesized, and the activity against hepatocellular carcinoma Huh-7 and HepG2 cells was evaluated. K&A DNA/RNA H-8 synthesizer was used to synthesize siRNAs, and neutral cytidinyl lipid DNCA mixed with cationic lipid CLD were used to transfect siRNA. By RT-qPCR and CCK-8 assay, the target gene silence and the proliferation of Huh-7 and HepG2 cells were detected. The siRNAs loading into Ago2 protein was detected by RNA-binding protein immunoprecipitation. Drug uptake and cell apoptosis were detected by flow cytometry, and the expression of PLK1 protein was detected by Western blot. Partial full 2ʹ-F/OMe modified siRNAs, especial siPLK1A3, increased the uptake of Huh-7 cells, enhanced their binding to Ago2 and gene silencing activity, down-regulated PLK1 protein, as well as induced more Huh-7 cell apoptosis and proliferation inhibition activity. It provides important data for the development of novel siRNA modification patterns and anti-HCC formulations.
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A dry suspension of Indigo Naturalis (IN) based on lactose-IN composite particles was designed by powder modification technology to meet the clinical needs of IN. The contact angle was used as an evaluation index to investigate the effects of the type of modifier lactose, the amount of lactose, and the co-grinding time of lactose and IN on the hydrophilicity of IN. The difference between IN before and after modification was compared through physical properties such as particle size and scanning electron microscope, as well as hydrophilic properties such as surface free energy and multiple light scattering. The optimal process of lactose-IN composite particles is as follows: after lactose is ground alone for 2 minutes, it is co-ground with IN at a ratio of 1∶1 for 6 minutes. The results of the investigation of powder properties show that the particle size d0.9 of IN is reduced from 112.75 μm to 87.30 μm after modification. The BET and Langmuir specific surface areas decreased by 8.661 m2·g-1 and 12.512 m2·g-1, respectively. SEM shows that lactose is attached to the surface of modified IN (MIN); surface element analysis shows that Si, Ca, and Mg elements of MIN are smaller than IN, and O elements are larger. The infrared spectrum shows that the MIN possesses the characteristic peaks of both IN and lactose. Compared MIN with IN, the contact angle and the non-polar surface free energy decreased by 35.1° and 9.975 mJ·m-2, respectively; the polar surface free energy and the surface free energy increased by 36.956 and 26.950 mJ·m-2, respectively. The results of multiple light scattering showed that the light transmittance of MIN was 35% lower than that of IN, and the backscattered light intensity was increased by about 25%. Only one excipient was used to successfully prepare IN dry suspension with good wettability and suspending property, which provided a basis for the development of new preparations of IN.
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Objective@#To investigate GATA3 expression and the regulatory mechanism of m6A modification in the re- sponse of alveolar epithelial cells to radiation, and to provide a new therapeutic target for radiation-induced lung injury based on its pathogenesis.@*Methods@#Human lung epithelial cell line (A549) and mouse lung epithelial cell line (MLE-12) were exposed to X-ray irradiation with a single dose of 10 Gy (dose rate 1 Gy/min) and 6 Gy (dose rate 0.75 Gy/min), respect- ively. The expression of VIRMA gene (RNA methylase) was inhibited by lipofection of A549 cells and MLE-12 cells with shRNA-VIRMA plasmid and siRNA-VIRMA interfering fragment, respectively. Quantification of m6A RNA methylation was performed by colorimetry. Changes in the expression of mRNAs of VIRMA, GATA3, and epithelial-mesenchymal transition (EMT) markers in irradiated A549 and MLE-12 cells were determined by qRT-PCR. Changes in the expression of VIRMA, GATA3, and EMT marker proteins in irradiated A549 and MLE-12 cells were determined by Western blot.@*Results@#Radiation up-regulated the expression of methylase VIRMA in A549 and MLE-12 cells, which in turn enhanced the m6A of total RNA and the expression of GATA3 gene and protein, resulting in EMT. Furthermore, in A549 and MLE-12 cells, interference of the VIRMA gene significantly reduced the expression of GATA3 gene and protein and the expression of EMT-related molecules.@*Conclusion @#Radiation induces m6A modification in alveolar epithelial cells, which up-regu- lates the expression of GATA3 gene and induces EMT, thus playing an important role in the process of radiation-induced lung injury.
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@#At present,malignant tumor has become one of the public problems that seriously threaten human health. In addition to surgery,radiotherapy,chemotherapy,targeted therapy and other methods,with the development of molecular biology,immunotherapy has also developed rapidly,becoming an emerging method of cancer treatment. The most commonly used immune cells in clinical treatment are DC,NK,CIK,CTL and chimeric antigen receptor T cell(CAR-T). Among them,CAR-T technology is the initial technology for global research,while due to its off-target,neurotoxicity,transfection vector defects and other problems,it also has certain limitations in clinical application. T cell antigen coupler modified T cell(TAC-T)technology is a new technology developed on the basis of CAR-T,which uses natural T cell receptor(TCR)to modify T cells and retarget the antigen of cancer cells. In this paper,the research status of CAR-T technology and the research progress of TAC-T technology are reviewed in order to provide reference for further study on the mechanism of TAC-T technology and its safety of clinical application.
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@#Objective To optimize the expression of recombinant human growth hormone-Fc(rhGH-Fc)fusion protein in CHO cells in order to obtain better glycosylation ratio and lower content of highmannose.Methods CHO cells expressing rhGH-Fc were cultured in a 7 L bioreactor.The glycosylation modifications of rhGH-Fc were adjusted by improving the composition of feeding media(using three commercial media:Gly-1:EX-CELL Glycosylation Adjust,Gly-2:SHEFF-CHO PLUS PG ACF and Gly-3:EfficientFeed C + AGT Supplement & GlycanTune C + Total Feed),and the glycosylation type and proportion of the target proteins were analyzed by mass spectrometry.Results The G0F(main glycosylation types:G0,G1 and G2;F:fucose)of Gly-1,Gly-2 and Gly-3 were 32.89%,58.66% and 33.28%,the G1F were 31.39%,18.03%and 34.90%,and the G2F were 31.39%,18.03% and 34.90%,respectively.Gly-1 and Gly-3 made the target protein contain less G0F while more G2F;Gly-3 feeding scheme-showed less high mannose modification than the other two schemes.Conclusion Gly-1 medium changed the glycosylation modification from G0F to G1F and G2F,while Gly-2 medium changed that from G2F and G1F to G0F.However,Gly-3 medium changed the glycosylation modification from G0F to G1F and G2F,and the contentof high mannose was less than 5%,which may have a better effect on modifying glycosylation type and proportion of the target protein.
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As a type of extracellular vesicles, exosomes are released by living cells and contain diverse bioactive molecules, including nucleic acids, proteins, lipids and metabolites. They play an important role in various physiological and pathological processes by a special intercellular communication medium. As endogenous vesicles, exosomes also have the advantages of systemic circulation stability, good biocompatibility and specific targeting of tissues and cells, as well as they are promising candidates for drug delivery system. The production mechanism of exosomes describe was summarized, the methods of extraction and separation the application and mechanism of exosomes in immune and inflammation-related diseases, cardiovascular system diseases, nervous system diseases, tumors, etc. were reviewed. The engineering modifications of exosomes in high targeting properties based on the drug delivery were overviewed. Exosomes support the diagnosis and prognostic assessment of multiple diseases, which have broad application prospects as a very potential safe and specific endogenous nano-drug carrier.
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Exosome is a kind of vesicle secreted by a variety of cells with lipid bilayer membrane structure， which has good biocompatibility， high targeting and high stability， and is a natural nanoscale drug carrier with great development potential in drug delivery system. In this paper， exosomes and their properties， exosome drug delivery pathways and methods， the design strategy of engineered exosome drug delivery systems for targeted disease therapy， and the application of exosome drug delivery systems in the treatment of a variety of diseases were reviewed. Exosome drug delivery pathways could be divided into two categories： exogenous and endogenous. Common exosome drug delivery methods included electroporation， co-incubation， and ultrasound. Engineered exosome drug delivery system can further improve drug loading and enhance drug targeting. The main way of engineering is to modify exosome surface through genetic engineering technology， physical modification， chemical modification， etc. Exosome drug delivery system provides a new idea for targeted therapy of arthritis， tumor， brain and other diseases.
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AIM: To investigate the role and mechanism of methyltransferase-like 3(METTL3)-mediated N6-methyladenosine(m6A)methylation modification in regulating biological activity of vascular endothelial cells in the pathogenesis of choroidal neovascularization.METHODS: Human umbilical vein endothelial cells(HUVEC)cultured in vitro were divided into the following groups: control group(normal culture), low density lipoprotein(LDL)group, fluorescence-labelled LDL(Dil-LDL)group, 12.5μg/mL and 25μg/mL oxidized LDL(ox-LDL)groups, 12.5μg/mL and 25μg/mL fluorescence-labelled ox-LDL(Dil-ox-LDL)groups, DMSO group, STM2457(METTL3 inhibitor)group, DAPT group; and monkey retina-choroidal endothelial cells(RF/6A)cultured in vitro were divided into control group, DMSO group, 12.5 μg/mL ox-LDL group, and DAPT group. Endocytosed lipoprotein level was examined through fluorescence microscopy. RNA m6A methylation level was detected through a dot blot assay. Protein and RNA levels of METTL3 or angiogenesis-related markers were measured through Western blot assays and real-time quantitative polymerase chain reaction(RT-qPCR), respectively. METTL3 expression and localization were investigated through immunofluorescence. Cell migratory and tube formation capacities were assessed through transwell migration and tube formation assays, respectively.RESULTS: Endocytosed lipoprotein levels in HUVECs exposed to Dil-LDL, 12.5μg/mL and 25μg/mL Dil-ox-LDL groups were significantly higher than those in the control group. 12.5μg/mL and 25μg/mL ox-LDL groups significantly increased m6A methylation(all P<0.05), METTL3 protein expression(all P<0.01), and cell migration and angiogenesis capacities(all P<0.01). METTL3 mRNA level was significantly unregulated in the 12.5μg/mL ox-LDL group(P<0.05). In comparison to the DMSO group, the addition of STM2457 caused significant decrease in m6A methylation level(P<0.05), expression of VEGF and other angiogenesis-related markers(all P<0.05), cell migration and angiogenesis capacities(all P<0.01)and the expression of NICD(P<0.05). However, there were no significant differences in METTL3 protein and mRNA levels(all P>0.05). The expression of VEGF and NICD(all P<0.05), as well as the ability of cell migration and angiogenesis of RF/6A, was all significantly decreased in the DAPT group compared to the DMSO group(all P<0.01).CONCLUSION: METTL3-mediated m6A methylation modification promotes angiogenesis in vascular endothelial cells via the Notch signaling pathway in the pathogenesis of choroidal neovascularization.
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@#Congenital cleft lip and/or palate (CL/P) is a common malformation of maxillofacial development. At present, it is believed that the etiology of congenital cleft lip and palate mainly results from genetic factors and environmental factors. Epigenetic changes induced by environmental factors may be the key factor in the occurrence of fetal congenital malformations. As one of the important epigenetic modifications, DNA methylation has been widely and deeply studied in many fields, but as a link between the individual and the environment, its application in CL/P is limited. Existing studies have shown that DNA methylation is closely related to the occurrence of cleft lip and palate. Stimulation of folate deficiency, smoking, pollutant exposure and other environmental factors can induce changes in the state of DNA methylation, thus affecting gene expression in the development of lip and palate and leading to the occurrence of deformities.