RESUMO
To investigate the effects of levothyroxine combined with methimazole on the clinical efficacy of hyperthyroidism treatment. A total of 102 patients with hyperthyroidism admitted to our hospital from January 2018 to June 2020 were selected and randomly assigned into the combination group (levothyroxine combined with methimazole) and the control group (methimazole treatment alone). 3 months after treatment, the two groups were compared with regard to clinical efficacy, changes in ultrasound findings, the thyroid hormones, and serum indexes and the adverse reactions rate. The combination group (98.04%) outperformed the control group (86.27%) in total effective rate, and the overall efficacy garnered the similar result. After treatment, the combination group showed advantages in thyroid hormone level, serum index level, thyroid volume, superior thyroid artery diameter, and maximum blood flow rate when compared with those of the control group (P<0.05). As for the adverse reactions rate, the combination group was superior to the control group (3.92%vs15.69%) (P<0.05). Levothyroxine combined with methimazole promotes the clinical efficacy of hyperthyroidism treating, reduces thyroid volume and the diameter of superior thyroid artery, enhances the patient's thyroid function and serum index, with higher safety profile.
Assuntos
Hipertireoidismo/tratamento farmacológico , Metimazol/uso terapêutico , Tiroxina/uso terapêutico , Adulto , Idoso , Antitireóideos/administração & dosagem , Antitireóideos/uso terapêutico , Quimioterapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tiroxina/administração & dosagem , Adulto JovemRESUMO
In this study, the mineral-weathering bacterium Pseudomonas azotoformans F77, which was isolated from the soil of a debris flow area, was evaluated for its weathering activity under direct contact with biotite or without contact. Then, biotite-weathering behaviors of strain F77, mutants that had been created by deleting the gcd and adh genes (which are involved in gluconic acid metabolism and pilus formation, respectively), and the double mutant F77ΔgcdΔadh were compared. The relative gene expression levels of F77 and its mutants F77Δgcd and F77Δadh were also analyzed in the presence of biotite. Direct contact with biotite increased Fe and Al release from the mineral in the presence of F77. All strains had similar abilities to release Fe and Al from the mineral except for F77Δgcd and F77Δadh Mobilized Fe and Al concentrations were decreased by up to 72, 26, and 87% in the presence of F77Δgcd, F77Δadh, and F77ΔgcdΔadh, respectively, compared to levels observed in the presence of F77 during the mineral-weathering process. Gluconic acid production was decreased for F77Δgcd and F77ΔgcdΔadh, while decreased cell attachment on the mineral surface was observed for F77Δadh, compared to findings for F77. The F77 genes involved in pilus formation and gluconic acid metabolism showed increased expression levels in the presence of biotite. The results of this study showed important roles for the genes involved in gluconic acid metabolism and pilus formation in mineral weathering by F77 and demonstrated the distinctive effect of these genes on mineral weathering by F77.IMPORTANCE Bacteria play important roles in mineral weathering and soil formation, although the molecular mechanisms underlying the interactions between bacteria and silicate minerals are poorly understood. In this study, the interactions between biotite and the highly effective mineral-weathering bacterium P. azotoformans F77 were characterized. Our results showed that the genes involved in gluconic acid metabolism and pilus formation play important roles in mineral weathering by F77. The presence of biotite could promote the expression of these genes in F77, and a distinctive effect of these genes on mineral weathering by F77 was observed in this study. Our results provide new knowledge and promote better understanding regarding the interaction between silicate minerals and mineral-weathering bacteria, as well as the molecular mechanisms involved in these processes.
Assuntos
Silicatos de Alumínio/metabolismo , Compostos Ferrosos/metabolismo , Minerais/metabolismo , Pseudomonas/metabolismo , Microbiologia do SoloRESUMO
Cell-to-cell communication precisely controls the creation of new organs during reproductive growth. However, the sensor molecules that mediate developmental signals in monocot plants are poorly understood. Here, we report that DWARF AND RUNTISH SPIKELET1 (DRUS1) and DRUS2, two closely related receptor-like kinases (RLKs), redundantly control reproductive growth and development in rice (Oryza sativa). A drus1-1 drus2 double knockout mutant, but not either single mutant, showed extreme dwarfism and barren inflorescences that harbored sterile spikelets. The gibberellin pathway was not impaired in this mutant. A phenotypic comparison of mutants expressing different amounts of DRUS1 and 2 revealed that reproductive growth requires a threshold level of DRUS1/2 proteins. DRUS1 and 2 maintain cell viability by repressing protease-mediated cell degradation and likely by affecting sugar utilization or conversion. In the later stages of anther development, survival of the endothecium requires DRUS1/2, which may stimulate expression of the UDP-glucose pyrophosphorylase gene UGP2 and starch biosynthesis in pollen. Unlike their Arabidopsis thaliana ortholog FERONIA, DRUS1 and 2 mediate a fundamental signaling process that is essential for cell survival and represents a novel biological function for the CrRLK1L RLK subfamily.
Assuntos
Metabolismo dos Carboidratos/genética , Oryza/genética , Proteínas de Plantas/genética , Receptores Proteína Tirosina Quinases/genética , Sequência de Aminoácidos , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Morte Celular/genética , Flores/enzimologia , Flores/genética , Flores/ultraestrutura , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas , Immunoblotting , Hibridização In Situ , Microscopia Confocal , Microscopia Eletrônica , Oryza/enzimologia , Fosfotransferases/genética , Fosfotransferases/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Interferência de RNA , Receptores Proteína Tirosina Quinases/metabolismo , Reprodução/genética , Homologia de Sequência de Aminoácidos , Amido/metabolismoRESUMO
BACKGROUND: Previous studies have suggested that patients with diabetes mellitus (DM) have higher prevalence of atherosclerotic cardiovascular disease (ASCVD), and plasma levels of free fatty acids (FFAs) are a useful marker for predicting ASCVD. We hypothesized that FFAs could predict both coronary and carotid lesions in an individual with type 2 DM (T2DM). The present study, hence, was to investigate the relation of plasma FFA level to the presence and severity of coronary and carotid atherosclerosis in patients with T2DM. METHODS: Three hundred and two consecutive individuals with T2DM who have received carotid ultrasonography and coronary angiography due to chest pain were enrolled in this study. Plasma FFAs were measured using an automatic biochemistry analyzer. Coronary and carotid severity was evaluated by Gensini score and Crouse score respectively. Subsequently, the relation of FFA levels to the presence and severity of coronary artery disease (CAD) and carotid atherosclerotic plaque (CAP) in whole individuals were also assessed. RESULTS: Increased plasma FFA levels were found in the groups either CAD or CAP compared to those without. Patients with higher level of FFAs had a higher CAD (89.9%) and elevated prevalence of CAP (69.7%). And also, patients with higher level of FFAs had a higher Gensini and Crouse scores. Multivariate regression analysis showed that FFA levels were independently associated with the presence of CAD and CAP (OR = 1.83, 95%CI: 1.27-2.65, P = 0.001; OR = 1.62, 95%CI: 1.22-2.14, P = 0.001, respectively). The area under the curve (AUC) was 0.68 and 0.65 for predicting the presence of CAD and CAP in patients with DM respectively. CONCLUSIONS: The present study firstly indicated that elevated FFA levels appeared associated with both the presence and severity of CAD and CAP in patients with T2DM, suggesting that plasma FFA levels may be a useful biomarker for improving management of patients with T2DM.
Assuntos
Biomarcadores/sangue , Doenças das Artérias Carótidas/diagnóstico , Doença da Artéria Coronariana/diagnóstico , Diabetes Mellitus Tipo 2/complicações , Ácidos Graxos não Esterificados/sangue , Índice de Gravidade de Doença , Doenças das Artérias Carótidas/sangue , Doenças das Artérias Carótidas/etiologia , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/etiologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
OBJECTIVES: In order to continuously promote the rational clinical application of antibacterial drugs and provide data support for hospital scientific management decision-making, we evaluated the use rate of antibiotics in outpatients in 2010 - 2018. MATERIALS AND METHODS: A retrospective analysis of the use rate of antibiotics in outpatients from January 2010 to October 2018 was performed in our hospital. We analyzed the antibacterial use rate in each month in outpatients and evaluated the changes in the antibacterial use rate in outpatients over the past 9 years. Autoregressive integrated moving average (ARIMA) model was established and verified, and we predicted the trends of antibacterial use rate in outpatients from November 2018 to March 2019. RESULTS: The level of clinical rational drug use management in our hospital is continually improving. The average monthly use rate of antibiotic drugs in outpatients in 2010 - 2013 decreased from 13.04 to 12.28% in 2014 - 2018 (p = 0.002). The goodness-of-fit of the ARIMA model is high. The verification results show that the model can accurately predict the changing trend. The average absolute error of the actual value and the fitted value of the antibiotic drug use rate was 0.76% from January to October 2018. The established ARIMA model can better simulate the trend of antibacterial use rate in outpatients. CONCLUSION: The model can be applied to the short-term prediction and dynamic analysis of antibacterial use rate in outpatients, helping hospitals to reasonably evaluate and monitor the use of clinical antibiotics and provide decision-making services for hospital management.
Assuntos
Antibacterianos/uso terapêutico , Prescrições de Medicamentos/estatística & dados numéricos , Modelos Estatísticos , Pacientes Ambulatoriais , China , Previsões , Humanos , Incidência , Estudos RetrospectivosRESUMO
Bacteria play important roles in mineral weathering and soil formation. However, little is known regarding the interactions between biotite and Arthrobacter strains. In this study, the mineral-mineral activities of the Arthrobacter pascens F74 isolated from a weathered rock surface were evaluated for its weathering behavior under direct contact and no contact with biotite. No contact was obtained by using dialysis bags. When directly in contact with biotite, Al and Fe concentrations increased by 9- to 47-fold compared with the controls in the presence of strain F74. Furthermore, strain F74 increased mobilized Al by 106% to 175% and Fe by 29% to 123% under direct contact than under no contact conditions. During biotite dissolution, significantly higher cell numbers and lower pH in the culture medium were observed in the presence of strain F74 under direct contact conditions than under no contact conditions. Significantly higher gluconic acid concentration and glucose dehydrogenase activity were found under direct contact conditions than under no contact and no biotite conditions. Scanning electron microscopy analysis showed cell adhesion on the biotite surface. These results demonstrated that strain F74 behaved differently with respect to biotite-weathering effectiveness and mechanisms under different contact conditions. The results also suggested that direct contact between biotite and strain F74 was important for the production of gluconic acid, cell adhesion on the mineral surface, and the mineral dissolution of the strain.
Assuntos
Silicatos de Alumínio/química , Arthrobacter/metabolismo , Compostos Ferrosos/química , Gluconatos/análise , Minerais/química , Alumínio/química , Glucose 1-Desidrogenase/metabolismo , Concentração de Íons de Hidrogênio , Ferro/química , Microscopia Eletrônica de Varredura , Microbiologia do SoloRESUMO
BACKGROUND: High-molecular-weight glutenin subunits (HMW-GS) play important roles in the elasticity of dough made from wheat. The HMW-GS null line is useful for studying the contribution of HMW-GS to the end-use quality of wheat. METHODS: In a previous work, we cloned the Glu-1Ebx gene from Thinopyrum bessarabicum and introduced it into the wheat cultivar, Bobwhite. In addition to lines expressing the Glu-1Ebx gene, we also obtained a transgenic line (LH-11) with all the HMW-GS genes silenced. The HMW-GS deletion was stably inherited as a dominant and conformed to Mendel's laws. Expression levels of HMW-GS were determined by RT-PCR and epigenetic changes in methylation patterns and small RNAs were analyzed. Glutenins and gliadins were separated and quantitated by reversed-phase ultra-performance liquid chromatography. Measurement of glutenin macropolymer, and analysis of agronomic traits and end-use quality were also performed. RESULTS: DNA methylation and the presence of small double-stranded RNA may be the causes of post-transcriptional gene silencing in LH-11. The accumulation rate and final content of glutenin macropolymer (GMP) in LH-11 were significantly lower than in wild-type (WT) Bobwhite. The total protein content was not significantly affected as the total gliadin content increased in LH-11 compared to WT. Deletion of HMW-GS also changed the content of different gliadin fractions. The ratio of ω-gliadin increased, whereas α/ß- and γ-gliadins declined in LH-11. The wet gluten content, sedimentation value, development time and stability time of LH-11 were remarkably lower than that of Bobwhite. Bread cannot be made using the flour of LH-11. CONCLUSIONS: Post-transcriptional gene silencing through epigenetic changes and RNA inhibition appear to be the causes for the gene expression deficiency in the transgenic line LH-11. The silencing of HMW-GW in LH-11 significantly reduced the dough properties, GMP content, wet gluten content, sedimentation value, development time and stability time of flour made from this wheat cultivar. The HMW-GS null line may provide a potential material for biscuit-making because of its low dough strength.
Assuntos
Pão , Glutens/metabolismo , Proteínas de Plantas/metabolismo , Triticum/metabolismo , Pão/normas , Culinária , Metilação de DNA , Farinha , Glutens/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Interferência de RNA , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Triticum/genéticaRESUMO
The plastidic glutamine synthetase isoform (GS2) plays a key role in nitrogen (N) assimilation. We introduced TaGS2-2Abpro::TaGS2-2Ab, the favourable allele of TaGS2-2A in the winter wheat (Triticum aestivum) variety Ji5265. Transgenic expression of TaGS2-2Ab significantly increased GS2 abundance and GS activity in leaves. Two consecutive field experiments showed the transgenic lines had higher grain yield, spike number, grain number per spike and 1000-grain weight than did the wild type under both low N and high N conditions. Analysis of N use-related traits showed that transgenic expression of TaGS2-2Ab increased root ability to acquire N, N uptake before and after flowering, remobilization of N to grains and N harvest index. Measurement of chlorophyll content and net photosynthesis rate in flag leaves during grain filling stage revealed that the transgenic lines had prolonged leaf functional duration as compared with the wild type. These results suggest that TaGS2 plays important role in N use, and the favourable allele TaGS2-2Ab is valuable in engineering wheat with improved N use efficiency and grain yield.
Assuntos
Produção Agrícola , Glutamato-Amônia Ligase/genética , Nitrogênio/metabolismo , Plantas Geneticamente Modificadas/genética , Triticum/genética , Clorofila/metabolismo , Produção Agrícola/métodos , Glutamato-Amônia Ligase/metabolismo , Fotossíntese , Folhas de Planta/enzimologia , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Plastídeos/genética , Característica Quantitativa Herdável , Triticum/crescimento & desenvolvimento , Triticum/metabolismoRESUMO
Chronic kidney disease (CKD) and related inflammatory responses stimulate protein-energy wasting, a complication causing loss of muscle mass. Primarily, muscle wasting results from accelerated protein degradation via autophagic/lysosomal and proteasomal pathways, but mechanisms regulating these proteolysis pathways remain unclear. Since dephosphorylation of FoxOs regulates ubiquitin/proteasome protein metabolism, we tested whether a novel nuclear phosphatase, the small C-terminal domain phosphatase (SCP) 4, regulates FoxOs signaling and, in turn, muscle wasting. In cultured mouse myoblast cells, SCP4 overexpression stimulated proteolysis, while knockdown of SCP4 prevented the proteolysis stimulated by inflammatory cytokines. SCP4 overexpression led to nuclear accumulation of FoxO1/3a followed by increased expression of catabolic factors including myostatin, Atrogin-1, and MuRF-1, and induction of lysosomal-mediated proteolysis. Treatment of C2C12 myotubes with proinflammatory cytokines stimulated SCP4 expression in an NF-κB-dependent manner. In skeletal muscle of mice with CKD, SCP4 expression was up-regulated. Similarly, in skeletal muscle of patients with CKD, SCP4 expression was significantly increased. Knockdown of SCP4 significantly suppressed FoxO1/3a-mediated expression of Atrogin-1 and MuRF-1 and prevented muscle wasting in mice with CKD. Thus, SCP4 is a novel regulator of FoxO transcription factors and promotes cellular proteolysis. Hence, targeting SCP4 may prevent muscle wasting in CKD and possibly other catabolic conditions.
Assuntos
Fatores de Transcrição Forkhead/metabolismo , Músculo Esquelético/enzimologia , Fosfoproteínas Fosfatases/metabolismo , Insuficiência Renal Crônica/complicações , Síndrome de Emaciação/etiologia , Animais , Humanos , Masculino , Camundongos Endogâmicos C57BL , Proteínas Musculares/metabolismo , Proteólise , Insuficiência Renal Crônica/enzimologia , Proteínas Ligases SKP Culina F-Box/metabolismo , Proteínas com Motivo Tripartido/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Síndrome de Emaciação/enzimologiaRESUMO
Skeletal muscle atrophy is an important clinical characteristic of chronic kidney disease (CKD); however, at present, the therapeutic approaches to muscle atrophy induced by CKD are still at an early stage of development. Resveratrol is used to attenuate muscle atrophy in other experimental models, but the effects on a CKD model are largely unknown. Here, we showed that resveratrol prevented an increase in MuRF1 expression and attenuated muscle atrophy in vivo model of CKD. We also found that phosphorylation of NF-κB was inhibited at the same time. Dexamethasone-induced MuRF1 upregulation was significantly attenuated in C2C12 myotubes by resveratrol in vitro, but this effect on C2C12 myotubes was abrogated by a knockdown of NF-κB, suggesting that the beneficial effect of resveratrol was NF-κB dependent. Our findings provide novel information about the ability of resveratrol to prevent or treat muscle atrophy induced by CKD.
Assuntos
Proteínas Musculares/metabolismo , Atrofia Muscular/metabolismo , Atrofia Muscular/prevenção & controle , Insuficiência Renal Crônica/tratamento farmacológico , Insuficiência Renal Crônica/metabolismo , Estilbenos/administração & dosagem , Proteínas com Motivo Tripartido/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Animais , Relação Dose-Resposta a Droga , Camundongos , Camundongos Endogâmicos C57BL , Atrofia Muscular/etiologia , NF-kappa B/metabolismo , Insuficiência Renal Crônica/complicações , Resveratrol , Transdução de Sinais/efeitos dos fármacosRESUMO
As a potential hemoglobin (Hb)-based oxygen carrier (HBOC), the PEGylated Hb has received much attention for its non-nephrotoxicity. However, PEGylation can adversely alter the structural and functional properties of Hb. The site of PEGylation is an important factor to determine the structure and function of the PEGylated Hb. Thus, protection of some sensitive residues of Hb from PEGylation is of great significance to develop the PEGylated Hb as HBOC. Here, Cys-93(ß) of Hb was conjugated with 20 kDa polyethylene glycol (PEG20K) through hydrazone and disulfide bonds. Then, the conjugate was modified with PEG5K succinimidyl carbonate (PEG5K-SC) using acylation chemistry, followed by removal of PEG20K Hb with hydrazone hydrolysis and disulfide reduction. Reversible conjugation of PEG20K at Cys-93(ß) can protect Lys-95(ß), Val-1(α) and Lys-16(α) of Hb from PEGylation with PEG5K-SC. The autoxidation rate, oxygen affinity, structural perturbation and tetramer instability of the PEGylated Hb were significantly decreased upon protection with PEG20K. The present study is expected to improve the efficacy of the PEGylated Hb as an oxygen therapeutic.
Assuntos
Cisteína/química , Hemoglobina A/química , Hemoglobina A/metabolismo , Oxigênio/metabolismo , Polietilenoglicóis/química , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Dicroísmo Circular , UltracentrifugaçãoRESUMO
Cytokinin plays an important role in plant development and stress tolerance. Studies of Arabidopsis (Arabidopsis thaliana) have demonstrated that cytokinin acts through a two-component system that includes a histidine (His) kinase, a His phosphotransfer protein (HP), and a response regulator. Phylogenetic analyses have revealed the conservation of His kinases but lineage-specific expansion of HPs and response regulators in rice (Oryza sativa). However, whether the functions of rice HPs have diverged remains unknown. In this study, two rice authentic HPs (OsAHP1 and OsAHP2) were knocked down simultaneously via RNA interference (RNAi), and the transgenic OsAHP-RNAi plants exhibited phenotypes expected for a deficiency in cytokinin signaling, including dwarfism with reduced internode lengths, enhanced lateral root growth, early leaf senescence, and reduced tiller numbers and fertility under natural conditions. The OsAHP-RNAi seedlings were also hyposensitive to exogenous cytokinin. Furthermore, OsAHP-RNAi seedlings were hypersensitive to salt treatment but resistant to osmotic stress relative to wild-type plants. These results indicate that OsAHPs function as positive regulators of the cytokinin signaling pathway and play different roles in salt and drought tolerance in rice.
Assuntos
Citocininas/metabolismo , Oryza/metabolismo , Oryza/fisiologia , Transdução de Sinais , Estresse Fisiológico , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Citocininas/farmacologia , Citosol/efeitos dos fármacos , Citosol/metabolismo , Histidina , Oryza/genética , Oryza/crescimento & desenvolvimento , Pressão Osmótica/efeitos dos fármacos , Fenótipo , Fosforilação/efeitos dos fármacos , Desenvolvimento Vegetal/efeitos dos fármacos , Proteínas de Plantas , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/efeitos dos fármacos , Plantas Geneticamente Modificadas , Interferência de RNA/efeitos dos fármacos , Plântula/efeitos dos fármacos , Plântula/fisiologia , Transdução de Sinais/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Estresse Fisiológico/efeitos dos fármacosRESUMO
BACKGROUND: Atherosclerotic cardiovascular disease is one of the major diseases that seriously impacts human health. Combined drug therapy may be efficacious in delaying the occurrence of cardiovascular events. AIM: The current study was designed to investigate whether combined use of probucol (an anti-oxidant agent) with cilostazol (a platelet aggregation inhibitor) would increase the inhibitory effect of statins (a lipid-lowering agent) on atherosclerosis in moderately hypercholesterolemic rabbits. METHODS AND RESULTS: Thirty Japanese white rabbits were fed with a high cholesterol diet for 12 weeks, which was supplemented with either 0.005% atorvastatin alone or 0.005% atorvastatin plus 0.3% probucol and 0.3% cilostazol. Except for high-density lipoprotein cholesterol, no difference was found in plasma lipids among vehicle, statin, and the combined treatment group. However, atherosclerotic lesions were significantly reduced by statin treatment compared with vehicle. Moreover, we found that the anti-atherogenic effect of statin was further enhanced by the combined treatment, which was due to increased anti-inflammatory and anti-oxidant properties. CONCLUSIONS: These data demonstrated that combined drug treatment exhibits potent athero-protective effects via pleiotropic functions, such as anti-inflammatory and anti-oxidative stress, which is independent of the lipid-lowering effect.
Assuntos
Aterosclerose/complicações , Aterosclerose/tratamento farmacológico , Atorvastatina/uso terapêutico , Hipercolesterolemia/complicações , Hipercolesterolemia/tratamento farmacológico , Probucol/uso terapêutico , Tetrazóis/uso terapêutico , Animais , Aorta/efeitos dos fármacos , Aorta/patologia , Aterosclerose/sangue , Aterosclerose/patologia , Atorvastatina/farmacologia , Biomarcadores/metabolismo , Cilostazol , Quimioterapia Combinada , Hipercolesterolemia/sangue , Inflamação/patologia , Lipídeos/sangue , Masculino , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Probucol/farmacologia , Coelhos , Tetrazóis/farmacologiaRESUMO
OBJECTIVE: To investigate the effects of emodin on aquaporin 5 (AQP5) expression in rats with sepsis-induced acute lung injury. METHODS: We divided 60 adult male Sprague-Dawley rats, weighing 200-230 g, into four groups: control, sham surgery, model and emodin groups (n = 15 for each). We created a sepsis model with cecal ligation and puncture; the sham surgery group had their cecums replaced after exposure outside the abdominal cavity. Each group was further divided into three subgroups (n = 5 for each) and expressions of AQP5 mRNA and proteins in lung tissue were measured by real-time fluorescence polymerase chain reaction and western blot at 6,12 and 24 h after surgery. RESULTS: AQP5 expression did not change over time in the control group and sham surgery group, but decreased over time in the model group. The lowest expression was found in 12-h subgroup, which significantly differed from the 6-h subgroup (P < 0.01). Compared with the model group, AQP5 expression in the emodin group was significantly higher in all the subgroups (all P < 0.01). Expressions in the 12-h subgroup were the highest, and significantly differed from the other subgroups. We found that lung tissue damage, such as pulmonary edema, alveolar damage and the exudation of red blood cells in pulmonary interstitium and alveolar, was significantly milder in the emodin group under light microscope than the model group. CONCLUSION: AQP5 expression was significantly down-regulated in rats with sepsis-induced acute lung injury induced by cecal ligation and puncture. Early prophylactic use of emodin can significantly enhance the AQP5 expression, thus effectively reducing the degree of pulmonary edema in septic rats.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Lesão Pulmonar Aguda/genética , Aquaporina 5/genética , Emodina/administração & dosagem , Sepse/complicações , Lesão Pulmonar Aguda/etiologia , Lesão Pulmonar Aguda/metabolismo , Animais , Aquaporina 5/metabolismo , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Humanos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
PEGylation, including nonspecific and site-directed approaches, is a well-established and validated strategy to increase the stability, in vivo plasma retention time, and efficacy of protein pharmaceutics together with a reduction in immunogenicity and hydrophobicity. Site-directed conjugation by PEG-aldehyde is the most widely used method for N-terminal modification; however, the generation of multimodified products is inevitable because of lysine chemistry, which always leads to difficulties in purification and quantification. In this study, we developed a specific PEGylation strategy through the periodation of the N-terminus of interferon beta-1b (IFN-ß-1b) followed by the coupling of PEG-hydrazide. The prolonged elimination half-life and significantly diminished immunogenicity of the PEG-hydrazide-modified protein indicated the development of an effective process for improving the pharmacology and immunogenicity of IFN-ß-1b. We further conducted comparisons on the selectivity, velocity, yield, and pharmacokinetics of the two methods. The results demonstrated that the hydrazide-based conjugation was a highly specific coupling reaction that only produced homogeneous N-terminal mono-PEGylated conjugate but also generated heterogeneous multimodified products in the aldehyde-based process. In addition, a better PEGylation yield was found for the hydrazide conjugation compared with that of the aldehyde strategy. These investigations supply a practical approach for the site-specific modification of proteins with an N-terminal serine or threonine to achieve improved homogeneity of the conjugates as well as enhanced pharmacological properties.
Assuntos
Hidrazinas/química , Interferon beta/química , Polietilenoglicóis/química , Animais , Chlorocebus aethiops , Humanos , Interferon beta-1b , Interferon beta/biossíntese , Interferon beta/isolamento & purificação , Modelos Moleculares , Estrutura Molecular , Ratos , Células VeroAssuntos
Antígeno Carcinoembrionário/metabolismo , Neoplasias do Sistema Digestório/cirurgia , Linfonodo Sentinela/diagnóstico por imagem , Cirurgia Assistida por Computador/métodos , Neoplasias Colorretais/cirurgia , Neoplasias do Sistema Digestório/diagnóstico por imagem , Fluorescência , Humanos , Neoplasias Pancreáticas/cirurgia , Neoplasias Gástricas/cirurgiaRESUMO
BACKGROUND: 2-Phenylethanol (2-PE) is one of the most widely used spices. Recently, 2-PE has also been considered a potential aviation fuel booster. However, the lack of scientific understanding of the 2-PE biosynthetic pathway and the cellular response to 2-PE cytotoxicity are the most important obstacles to the efficient biosynthesis of 2-PE. RESULTS: Here, metabolic engineering and tolerance engineering strategies were used to improve the production of 2-PE in Komagataella phaffii. First, the endogenous genes encoding the amino acid permease GAP1, aminotransferase AAT2, phenylpyruvate decarboxylase KDC2, and aldehyde dehydrogenase ALD4 involved in the Ehrlich pathway and the 2-PE stress response gene NIT1 in K. phaffii were screened and characterized via comparative transcriptome analysis. Subsequently, metabolic engineering was employed to gradually reconstruct the 2-PE biosynthetic pathway, and the engineered strain S43 was obtained, which produced 2.98 g/L 2-PE in shake flask. Furthermore, transcriptional profiling analyses were utilized to screen for novel potential tolerance elements. Our results demonstrated that cells with knockout of the PDR12 and C4R2I5 genes exhibited a significant increase in 2-PE tolerance. To confirm the practical applications of these results, deletion of the PDR12 and C4R2I5 genes in the hyper 2-PE producing strain S43 dramatically increased the production of 2-PE by 18.12%, and the production was 3.54 g/L. CONCLUSION: This is the highest production of 2-PE produced by K. phaffii via L-phenylalanine conversion. These identified K. phaffii endogenous elements are highly conserved in other yeast species, suggesting that manipulation of these homologues might be a useful strategy for improving aromatic alcohol production. These results also enrich the understanding of aromatic compound biosynthetic pathways and 2-PE tolerance, and provide new elements and strategies for the synthesis of aromatic compounds by microbial cell factories.
RESUMO
The latest developments in deep learning have demonstrated the importance of CT medical imaging for the classification of pulmonary nodules. However, challenges remain in fully leveraging the relevant medical annotations of pulmonary nodules and distinguishing between the benign and malignant labels of adjacent nodules. Therefore, this paper proposes the Nodule-CLIP model, which deeply mines the potential relationship between CT images, complex attributes of lung nodules, and benign and malignant attributes of lung nodules through a comparative learning method, and optimizes the model in the image feature extraction network by using its similarities and differences to improve its ability to distinguish similar lung nodules. Firstly, we segment the 3D lung nodule information by U-Net to reduce the interference caused by the background of lung nodules and focus on the lung nodule images. Secondly, the image features, class features, and complex attribute features are aligned by contrastive learning and loss function in Nodule-CLIP to achieve lung nodule image optimization and improve classification ability. A series of testing and ablation experiments were conducted on the public dataset LIDC-IDRI, and the final benign and malignant classification rate was 90.6%, and the recall rate was 92.81%. The experimental results show the advantages of this method in terms of lung nodule classification as well as interpretability.
Assuntos
Neoplasias Pulmonares , Nódulo Pulmonar Solitário , Tomografia Computadorizada por Raios X , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/classificação , Neoplasias Pulmonares/patologia , Tomografia Computadorizada por Raios X/métodos , Nódulo Pulmonar Solitário/diagnóstico por imagem , Aprendizado Profundo , Pulmão/diagnóstico por imagem , Interpretação de Imagem Radiográfica Assistida por Computador/métodos , Bases de Dados FactuaisRESUMO
In this study, the Pb-resistant Ensifer adhaerens strain S24, which contains quorum sensing (QS) systems responsible for N-acyl homoserine lactone (AHL) production, was investigated for QS system-mediated Pb stabilization and the underlying mechanisms. Whole-genome sequence analysis revealed the QS SinI/R and TraI/R systems in strain S24. Subsequently, strains S24 and the S24∆sinI/R, S24∆traI/R, S24∆traI/R/sinR, and S24∆sinI/R-traI/R/sinR mutants were constructed and compared for QS SinI/SinR-TraI/TraR system-mediated Pb stabilization in the solution and the mechanisms involved. After 5 days of incubation, strain S24 significantly decreased the Pb concentration in the Pb-contaminated solution compared with the mutants. The S24∆sinI/R-traI/R/sinR mutant exhibited reduced Pb stabilization and AHL activity than the other mutants. The S24∆sinI/R-traI/R/sinR mutant had significantly greater Pb concentrations in the solution and lower cell surface-adsorbed and extracellular precipitated Pb (PbS) contents as well as lower expression of H2S-producing genes of metC and sseA than did strain S24. Furthermore, the S24∆sinI/R-traI/R/sinR mutant displayed reduced interactions between the hydroxyl, amino, carboxyl, and ether groups and Pb, compared with strain S24. These findings implied the vital role of the SinI/SinR-TraI/TraR systems in strain S24 for Pb stabilization through enhanced cell surface adsorption and extracellular precipitation in Pb-polluted aquatic environments.
RESUMO
In this study, the cadmium (Cd)-tolerant Ensifer adhaerens strain NER9 with quorum sensing (QS) systems (responsible for N-acyl homoserine lactone (AHL) production) was characterized for QS system-mediated Cd immobilization and the underlying mechanisms involved. Whole-genome sequence analysis revealed that strain NER9 contains the QS SinI/R and TraI/R systems. Strains NER9 and the NER9∆sinI/R, NER9∆traI/R, and NER9∆sinI/R-traI/R mutants were constructed and compared for QS SinI/R and TraI/R system-mediated Cd immobilization in the solution and the mechanisms involved. After 24 h of incubation, strain NER9 significantly decreased the Cd concentration in the Cd-contaminated solution compared with the NER9∆sinI/R, NER9∆traI/R, and NER9∆sinI/R-traI/R mutants. The NER9∆sinI/R mutant had a greater impact on Cd immobilization and a lower impact on the activities of AHLs than did the NER9∆traI/R mutant. The NER9∆sinI/R mutant had significantly greater Cd concentrations and lower cell wall- and exopolysaccharide (EPS)-adsorbed Cd contents than did strain NER9. Furthermore, the NER9∆sinI/R mutant presented a decrease in the number of functional groups interacting with Cd, compared with strain NER9. These results suggested that the SinI/R system in strain NER9 contributed to Cd immobilization by mediating cell wall- and EPS-adsorption in Cd-containing solution.