RESUMO
The CRISPR system is an adaptive immune system found in prokaryotes that defends host cells against the invasion of foreign DNA1. As part of the ongoing struggle between phages and the bacterial immune system, the CRISPR system has evolved into various types, each with distinct functionalities2. Type II Cas9 is the most extensively studied of these systems and has diverse subtypes. It remains uncertain whether members of this family can evolve additional mechanisms to counter viral invasions3,4. Here we identify 2,062 complete Cas9 loci, predict the structures of their associated proteins and reveal three structural growth trajectories for type II-C Cas9. We found that novel associated genes (NAGs) tended to be present within the loci of larger II-C Cas9s. Further investigation revealed that CbCas9 from Chryseobacterium species contains a novel ß-REC2 domain, and forms a heterotetrameric complex with an NAG-encoded CRISPR-Cas-system-promoting (pro-CRISPR) protein of II-C Cas9 (PcrIIC1). The CbCas9-PcrIIC1 complex exhibits enhanced DNA binding and cleavage activity, broader compatibility for protospacer adjacent motif sequences, increased tolerance for mismatches and improved anti-phage immunity, compared with stand-alone CbCas9. Overall, our work sheds light on the diversity and 'growth evolutionary' trajectories of II-C Cas9 proteins at the structural level, and identifies many NAGs-such as PcrIIC1, which serves as a pro-CRISPR factor to enhance CRISPR-mediated immunity.
Assuntos
Bactérias , Bacteriófagos , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Bactérias/virologia , Bactérias/genética , Bactérias/imunologia , Bacteriófagos/genética , Bacteriófagos/imunologia , Chryseobacterium/genética , Chryseobacterium/imunologia , Chryseobacterium/virologia , Proteína 9 Associada à CRISPR/química , Proteína 9 Associada à CRISPR/genética , Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Sistemas CRISPR-Cas/imunologia , Clivagem do DNA , Loci Gênicos/genética , Modelos Moleculares , Domínios ProteicosRESUMO
The emergence of CRISPR-Cas systems has accelerated the development of gene editing technologies, which are widely used in the life sciences. To improve the performance of these systems, workers have engineered and developed a variety of CRISPR-Cas tools with a broader range of targets, higher efficiency and specificity, and greater precision. Moreover, CRISPR-Cas-related technologies have also been expanded beyond making cuts in DNA by introducing functional elements that permit precise gene modification, control gene expression, make epigenetic changes, and so on. In this review, we introduce and summarize the characteristics and applications of different types of CRISPR-Cas tools. We discuss certain limitations of current approaches and future prospects for optimizing CRISPR-Cas systems.
Assuntos
Proteínas Associadas a CRISPR/genética , Sistemas CRISPR-Cas , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Edição de Genes , Animais , Proteínas Associadas a CRISPR/metabolismo , Difusão de Inovações , HumanosRESUMO
Cytosine base editors (CBEs) generate C-to-T nucleotide substitutions in genomic target sites without inducing double-strand breaks. However, CBEs such as BE3 can cause genome-wide off-target changes via sgRNA-independent DNA deamination. By leveraging the orthogonal R-loops generated by SaCas9 nickase to mimic actively transcribed genomic loci that are more susceptible to cytidine deaminase, we set up a high-throughput assay for assessing sgRNA-independent off-target effects of CBEs in rice protoplasts. The reliability of this assay was confirmed by the whole-genome sequencing (WGS) of 10 base editors in regenerated rice plants. The R-loop assay was used to screen a series of rationally designed A3Bctd-BE3 variants for improved specificity. We obtained 2 efficient CBE variants, A3Bctd-VHM-BE3 and A3Bctd-KKR-BE3, and the WGS analysis revealed that these new CBEs eliminated sgRNA-independent DNA off-target edits in rice plants. Moreover, these 2 base editor variants were more precise at their target sites by producing fewer multiple C edits.
Assuntos
Citidina Desaminase/genética , Citosina , Edição de Genes/métodos , Antígenos de Histocompatibilidade Menor/genética , Oryza/genética , Citosina/química , Genes de Plantas , Humanos , Mutação , RNA Guia de Cinetoplastídeos/química , RNA de Plantas/química , Reprodutibilidade dos TestesRESUMO
BACKGROUND: Venous thromboembolism (VTE) is a common postoperative complication in patients undergoing surgery for gastric cancer (GC). Although VTE incidence may vary among cancers, guidelines rarely stratify preventive methods for postoperative VTE by cancer type. The risk of VTE in patients undergoing surgery for GC remains unclear. METHODS: A systematic review and meta-analysis was undertaken to determine the risk of VTE after GC surgery and discuss the clinical value of pharmacological thromboprophylaxis in these cases. Medline, Embase, Web of Science, and Cochrane Library databases were searched for articles published from their inception to September 2022. RESULTS: Overall, 13 studies (111,936 patients) were included. The overall 1-month incidence of VTE, deep vein thrombosis (DVT), and pulmonary embolism (PE) after GC surgery was 1.8% (95% CI, 0.8-3.1%; I²=98.5%), 1.2% (95% CI, 0.5-2.1%; I²=96.1%), and 0.4% (95% CI, 0.1-1.1%; I²=96.3%), respectively. The prevalence of postoperative VTE was comparable between Asian and Western populations (1.8% vs. 1.8%; P > 0.05). Compared with mechanical prophylaxis alone, mechanical plus pharmacological prophylaxis was associated with a significantly lower 1-month rate of postoperative VTE and DVT (0.6% vs. 2.9% and 0.6% vs. 2.8%, respectively; all P < 0.05), but not PE (P > 0.05). The 1-month postoperative incidence of VTE was not significantly different between laparoscopic and open surgery (1.8% vs. 4.3%, P > 0.05). CONCLUSION: Patients undergoing GC surgery do not have a high risk of VTE. The incidence of VTE after GC surgery is not significantly different between Eastern and Western patients. Mechanical plus pharmacological prophylaxis is more effective than mechanical prophylaxis alone in postoperative VTE prevention. The VTE risk is comparable between open and laparoscopic surgery for GC.
Assuntos
Embolia Pulmonar , Neoplasias Gástricas , Tromboembolia Venosa , Humanos , Tromboembolia Venosa/epidemiologia , Tromboembolia Venosa/etiologia , Tromboembolia Venosa/prevenção & controle , Anticoagulantes/uso terapêutico , Neoplasias Gástricas/cirurgia , Neoplasias Gástricas/complicações , Embolia Pulmonar/epidemiologia , Embolia Pulmonar/etiologia , Embolia Pulmonar/prevenção & controle , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Fatores de RiscoRESUMO
BACKGROUND: The migration of lymphocytes shares many similarities in mode and mechanism with the metastasis of lung cancer tumor cells. But changes in the expression of lymphocyte migration regulation related proteins in urine exosomes remain unclear. This study is to investigate the expression changes of lymphocyte migration regulation related proteins in urine exosomes of lung cancer patients, and further verify their correlation with the development and progression of lung cancer. METHODS: Urine exosomes were collected from lung cancer patients and healthy people aged 15-79 years. Mass spectrometry was used to screen and explore the expression changes of lymphocyte migration regulation related proteins in healthy people of different ages. Enzyme-linked immunosorbent assay and western blotting were used to detect the expression changes of lymphocyte migration regulation related proteins in lung cancer patients. RESULTS: Analyzing the data of urine exosome proteomics, a total of 12 lymphocyte related proteins were identified, 5 of which were lymphocyte migration regulation related proteins. Among these proteins, WASL and STK10 proteins showed a gradual decrease in expression with age, and WNK1 protein showed a gradual increase. Lung cancer patients had reduced expression of WASL and increased expression of STK10 and WNK1 proteins in urine exosomes compared to normal people. Urine exosome WASL, STK10, and WNK1 were diagnosed with lung cancer, with a combined AUC of 0.760. CONCLUSIONS: Lymphocyte migration regulation related proteins were differentially expressed in the urine exosome of lung cancer patients, and WASL, STK10 and WNK1 may serve as potential biomarkers for lung cancer diagnosis.
Assuntos
Exossomos , Neoplasias Pulmonares , Humanos , Neoplasias Pulmonares/patologia , Exossomos/metabolismo , Pulmão/patologia , Biomarcadores/análise , Fatores de Transcrição/metabolismo , Proteínas Serina-Treonina Quinases/metabolismoRESUMO
Background Kidney renal clear cell carcinoma (KIRC) is one of the most common renal malignancies with a high mortality rate. Cuproptosis, a novel form of cell death, is strongly linked to mitochondrial metabolism and is mediated by protein lipoylation, leading to a proteotoxic stress response and cell death. To date, few studies have ellucidated the holistic role of cuproptosis-related genes (CRGs) in the pathogenesis of KIRC.Methods We comprehensively and completely analyzed the RNA sequencing data and corresponding clinical information from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. We screened for differentially expressed CRGs and constructed a prognostic risk model using univariate and multivariate Cox proportional regression analyses. Kaplan-Meier analysis was performed and receiver operating characteristic (ROC) curves were plotted to predict the prognosis of KIRC patients. Functional enrichment analysis was utilized to explore the internal mechanisms. Immune-related functions were analyzed using single-sample gene set enrichment analysis (ssGSEA), tumour immune dysfunction and exclusion (TIDE) scores, and drug sensitivity analysis.Results We established a concise prognostic risk model consisting of four CRGs (DBT, DLAT, LIAS and PDHB) to predict the overall survival (OS) in KIRC patients. The results of the survival analysis indicated a significantly lower OS in the high-risk group as compared to the patients in the low-risk group. The area under the time-dependent ROC curve (AUC) at 1, 3, and 5 year was 0.691, 0.618, and 0.614 in KIRC. Functional enrichment analysis demonstrated that CRGs were significantly enriched in tricarboxylic acid (TCA) cycle-related processes and metabolism-related pathways. Sorafenib, doxorubicin, embelin, and vinorelbine were more sensitive in the high-risk group.Conclusions We constructed a concise CRGs risk model to evaluate the prognosis of KIRC patients and this may be a new direction for the diagnosis and treatment of KIRC.
Assuntos
Apoptose , Carcinoma de Células Renais , Neoplasias Renais , Humanos , Carcinoma de Células Renais/tratamento farmacológico , Carcinoma de Células Renais/genética , Imunoterapia , Rim , Neoplasias Renais/tratamento farmacológico , Neoplasias Renais/genética , Prognóstico , CobreRESUMO
The monoamine oxidase-B (MAO-B) inhibitors with neuroprotective effects are better for Parkinson's disease (PD) treatment, due to the complicated pathogenesis of PD. To develop new hMAO-B inhibitors with neuroprotection, a novel series of 3,4-dihydrocoumarins was designed as selective and reversible hMAO-B inhibitors to treat PD. Most compounds showed potent and selective inhibition for hMAO-B over hMAO-A with IC50 values ranging from nanomolar to sub-nanomolar. Among them, compound 4d was the most potent hMAO-B inhibitor (IC50 = 0.37 nM) being about 20783-fold more active than iproniazid, and exhibited the highest selectivity for hMAO-B (SI > 270,270). Kinetic studies revealed that compound 4d was a reversible and competitive inhibitor of hMAO-B. Neuroprotective studies indicated that compound 4d could protect PC12 cells from the damage induced by 6-OHDA and rotenone. Besides, compound 4d did not exhibit acute toxicity at a dose up to 2500 mg/kg (po), and could cross the BBB in parallel artificial membrane permeability assay. More importantly, compound 4d was able to significantly prevent the motor deficits in the MPTP-induced PD model. These results indicate that compound 4d is an effective and promising candidate against PD.
Assuntos
Cumarínicos/química , Desenho de Fármacos , Intoxicação por MPTP/tratamento farmacológico , Inibidores da Monoaminoxidase/farmacologia , Monoaminoxidase/metabolismo , Animais , Indanos/farmacologia , Camundongos , Modelos Moleculares , Estrutura Molecular , Inibidores da Monoaminoxidase/química , Oxidopamina/toxicidade , Células PC12 , Conformação Proteica , Ratos , Rotenona/toxicidade , Relação Estrutura-AtividadeRESUMO
INTRODUCTION AND OBJECTIVE: The purpose of this study was to investigate the expression levels and prognostic roles of α-fetoprotein (AFP), carcinoembryonic antigen (CEA), and Ki67 in tumor tissues of intrahepatic cholangiocarcinoma (ICC) patients. PATIENTS OR MATERIALS AND METHODS: The study involved ninety-two ICC patients with complete clinicopathological data and follow-up information, who had previously undergone radical surgery. AFP, CEA, CD10, CD34, and Ki67 were detected in tumor tissues using immunohistochemistry. Statistical tests were used to identify independent risk factors and their associations with overall survival (OS) and disease-free survival (DFS). RESULTS: AFP, CEA and Ki67 were strongly correlated with prognosis. Univariate analysis indicated that higher AFP (Pâ¯=â¯0.002), CEA (Pâ¯<â¯0.0001), Ki67 (Pâ¯<â¯0.0001), CA19-9 (Pâ¯=â¯0.039), and CA12-5 (Pâ¯=â¯0.002), and larger tumor size (Pâ¯=â¯0.001), as well as more advanced tumor node metastasis (TNM) staging (Pâ¯<â¯0.0001) were all associated with worse OS. Meanwhile, higher AFP (Pâ¯=â¯0.002), CEA (Pâ¯=â¯0.001), and Ki67 (Pâ¯<â¯0.0001), as well as more advanced TNM staging (Pâ¯=â¯0.005) were associated with worse DFS. Multivariate analysis showed that higher AFP (HRâ¯=â¯2.004, 95%CI: 1.146-3.504 Pâ¯=â¯0.015), CEA (HRâ¯=â¯2.226, 95%CI: 1.283-3.861 Pâ¯=â¯0.004), and Ki67 (HRâ¯=â¯3.785, 95%CI: 2.073-6.909â¯Pâ¯<â¯0.0001), as well as more advanced TNM staging (HRâ¯=â¯2.900, 95%CI: 1.498-5.757 Pâ¯=â¯0.002) had associations with worse OS. Furthermore, higher AFP (HRâ¯=â¯2.172, 95%Cl: 1.291-3.654 Pâ¯=â¯0.003), CEA (HRâ¯=â¯1.934, 95%Cl: 1.180-3.169 Pâ¯=â¯0.009), and Ki67 (HRâ¯=â¯2.203, 95%Cl: 1.291-3.761 Pâ¯=â¯0.004) had associations with worse DFS. CONCLUSION: High AFP, CEA, and Ki67 are significant prognostic indicators in ICC patients, and can be used to evaluate ICC biological behavior and prognosis.
Assuntos
Neoplasias dos Ductos Biliares/sangue , Ductos Biliares Intra-Hepáticos , Antígeno Carcinoembrionário/sangue , Colangiocarcinoma/sangue , Antígeno Ki-67/sangue , alfa-Fetoproteínas/metabolismo , Idoso , Neoplasias dos Ductos Biliares/diagnóstico , Neoplasias dos Ductos Biliares/mortalidade , Biomarcadores/sangue , Colangiocarcinoma/diagnóstico , Colangiocarcinoma/mortalidade , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Taxa de SobrevidaRESUMO
BACKGROUND: The aim of this study was to investigate the prognostic value of arginase-1 (Arg-1) and glypican-3 (GPC-3) in patients with intrahepatic cholangiocarcinoma (ICC). METHODS: Two hundred and thirty-seven patients with ICC were included in this study. All patients had undergone radical surgery and had complete clinical information. Immunohistochemistry was used to assess the levels of Arg-1 and GPC-3 in ICC tissues. Univariate and multivariate analyses were conducted to identify independent risk factors in ICC. The relationship between Arg-1 and GPC-3 levels and patient survival was determined using the Kaplan-Meier method. RESULTS: High Arg-1 and GPC-3 expression levels were associated with poor prognosis in patients with ICC, and they could be as new prognostic biomarkers in ICC. CONCLUSION: Arg-1 and GPC-3 can serve as independent prognostic biomarkers in ICC.
Assuntos
Neoplasias dos Ductos Biliares , Colangiocarcinoma , Arginase , Neoplasias dos Ductos Biliares/cirurgia , Colangiocarcinoma/cirurgia , Glipicanas , Humanos , PrognósticoRESUMO
Androgen receptor (AR) can suppress hepatocellular carcinoma (HCC) invasion and metastasis at an advanced stage. Vasculogenic mimicry (VM), a new vascularization pattern by which tumour tissues nourish themselves, is correlated with tumour progression and metastasis. Here, we investigated the effect of AR on the formation of VM and its mechanism in HCC. The results suggested that AR could down-regulate circular RNA (circRNA) 7, up-regulate micro RNA (miRNA) 7-5p, and suppress the formation of VM in HCC Small hairpin circR7 (ShcircR7) could reverse the impact on VM and expression of VE-cadherin and Notch4 increased by small interfering AR (shAR) in HCC, while inhibition of miR-7-5p blocked the formation of VM and expression of VE-cadherin and Notch4 decreased by AR overexpression (oeAR) in HCC. Mechanism dissection demonstrated that AR could directly target the circR7 host gene promoter to suppress circR7, and miR-7-5p might directly target the VE-cadherin and Notch4 3'UTR to suppress their expression in HCC. In addition, knockdown of Notch4 and/or VE-cadherin revealed that shVE-cadherin or shNotch4 alone could partially reverse the formation of HCC VM, while shVE-cadherin and shNotch4 together could completely suppress the formation of HCC VM. Those results indicate that AR could suppress the formation of HCC VM by down-regulating circRNA7/miRNA7-5p/VE-Cadherin/Notch4 signals in HCC, which will help in the design of novel therapies against HCC.
Assuntos
Carcinoma Hepatocelular/etiologia , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/etiologia , Neoplasias Hepáticas/metabolismo , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Receptores Androgênicos/metabolismo , Antígenos CD/metabolismo , Biomarcadores , Caderinas/metabolismo , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/patologia , MicroRNAs/genética , Interferência de RNA , RNA Circular/genética , Receptor Notch4/metabolismo , Transdução de SinaisRESUMO
A series of new ferulic acid derivatives were designed, synthesized and evaluated as multi-target inhibitors against Alzheimer's disease. In vitro studies indicated that most compounds showed significant potency to inhibit self-induced ß-amyloid (Aß) aggregation and acetylcholinesterase (AChE), and had good antioxidant activity. Specifically, compound 4g exhibited the potent ability to inhibit cholinesterase (ChE) (IC50, 19.7â¯nM for hAChE and 0.66⯵M for hBuChE) and the good Aß aggregation inhibition (49.2% at 20⯵M), and it was also a good antioxidant (1.26 trolox equivalents). Kinetic and molecular modeling studies showed that compound 4g was a mixed-type inhibitor, which could interact simultaneously with the catalytic anionic site (CAS) and the peripheral anionic site (PAS) of AChE. Moreover, compound 4g could remarkably increase PC12 cells viability in hydrogen peroxide-induced oxidative cell damage and Aß-induced cell damage. Finally, compound 4g had good ability to cross the BBB using the PAMPA-BBB assay. These results suggested that compound 4g was a promising multifunctional ChE inhibitor for the further investigation.
Assuntos
Doença de Alzheimer/tratamento farmacológico , Anticoagulantes/uso terapêutico , Ácidos Cumáricos/química , Ácidos Cumáricos/síntese química , Simulação de Acoplamento Molecular/métodos , Doença de Alzheimer/patologia , Anticoagulantes/farmacologia , Desenho de Fármacos , Humanos , Ligantes , Modelos MolecularesRESUMO
Biothiols, including cysteine (Cys), homocysteine (Hcy), glutathione (GSH) and H2S, play important roles in human physiological processes. However, it is a great difficulty to distinguish biothiols from each other because of their similar chemical properties. Based on Nile red, we have designed and synthesized a near-infrared fluorescent probe for discriminating Cys/Hcy from GSH/H2S by a dual-channel detection method. Using an ether bond, near-infrared Nile red was attached to 7-nitrobenzofurazan to construct the probe. Due to the photo-induced electron transfer, the probe showed almost no fluorescence from the green to red emission band. But upon the addition of Cys (0-150 µM) or Hcy (0-200 µM), the probe exhibited a noteworthy fluorescence "turn-on" signal in two unique emission bands (Green and Red) with a fast response (within 30 min). In contrast, the probe displayed an increase in fluorescence only in the red channel when encountering GSH (0-70 µM) or H2S (0-50 µM), and GSH/H2S could be tested respectively by different response time. The limit of detection was calculated to be 0.09 µM (Cys), 0.30 µM (Hcy), 0.24 µM (GSH), and 0.04 µM (H2S), respectively (based on S/N = 3). The desirable dual-channel detection could be achieved in serum samples and living cells. Moreover, the probe could be applied for bioimaging in mice, which indicated its potential application in the clinic.
Assuntos
Cisteína/análise , Corantes Fluorescentes/química , Glutationa/análise , Homocisteína/análise , Sulfeto de Hidrogênio/análise , 4-Cloro-7-nitrobenzofurazano/análogos & derivados , 4-Cloro-7-nitrobenzofurazano/síntese química , 4-Cloro-7-nitrobenzofurazano/toxicidade , Animais , Linhagem Celular Tumoral , Fluorescência , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/toxicidade , Humanos , Concentração de Íons de Hidrogênio , Limite de Detecção , Camundongos Nus , Imagem Óptica/métodos , Oxazinas/síntese química , Oxazinas/química , Oxazinas/toxicidade , Espectrometria de FluorescênciaRESUMO
Based upon the intramolecular charge transfer (ICT) mechanism, a novel ratiometric fluorescent probe EB was developed to detect SO32-/HSO3-. The probe displayed both colorimetric and ratiometric responses toward SO32-/HSO3-. It displayed a quick response (within 60 s), good selectivity and high sensitivity (a detection limit of 28 nM) towards SO32-/HSO3-. The SO32-/HSO3- sensing mechanism was confirmed as the Michael addition reaction by ESI-MS. Moreover, the probe could be applied to measure the level of sulfite in real samples, like sugar and chrysanthemum, and it could also be used to detect SO32-/HSO3- in HepG2 cells through confocal fluorescence microscopy, which proved its practical application in clinical diagnosis.
Assuntos
Carbocianinas/química , Corantes Fluorescentes/química , Dióxido de Enxofre/química , Linhagem Celular Tumoral , Colorimetria/métodos , Fluorescência , Células Hep G2 , Humanos , Limite de Detecção , Sensibilidade e Especificidade , Sulfitos/químicaRESUMO
BACKGROUND: The goal of this meta-analysis is to explore the overall efficacy as well as the safety of anterior versus posterior approach for the therapy of patients with multilevel cervical spondylotic myelopathy based on qualified studies. METHODS: Three electronic databases, PubMed, Cochrane, Embase were searched updated to January 2018 to identify all relevant and qualified studies using the index words. The qualified studies were including prospective or retrospective comparative studies. Relative risk (RR) and mean difference (MD) along with 95% confidence interval (95% CI) were used to analyze the main outcomes. RESULTS: In this meta-analysis, there were a total of 24 studies with 959 patients in the anterior approach group and 1072 patients in the posterior approach group. The final results showed, in comparison of the posterior approach group, the anterior approach group significantly increased the JOA score (SMD: 0.36, 95% CI 0.10-0.62), the operation time (WMD: 49.87, 95% CI 17.67-82.08), and the neurological recovery rate (WMD: 10.55, 95% CI 3.99-17.11) with higher complication rate (RR: 1.53, 95% CI 1.24-1.89). Besides, there was no significant difference of the blood loss (SMD: - 0.40, 95% CI - 1.12 to 0.32) and ROM (SMD: - 0.28, 95% CI - 0.78 to - 0.22) between posterior approach group and anterior approach group. CONCLUSIONS: Studies found a significant increase of JOA score as well as neurological recovery rate by the anterior approach treatment when compared with posterior approach treatment. However, increased operation time and complications could also occur through the anterior approach treatment. More high-quality randomized controlled trials with larger sample size, multi-centric and longer follow-ups are needed to support our current conclusions.
Assuntos
Vértebras Cervicais/cirurgia , Procedimentos Ortopédicos , Doenças da Medula Espinal/cirurgia , Humanos , Duração da Cirurgia , Procedimentos Ortopédicos/efeitos adversos , Procedimentos Ortopédicos/métodos , Procedimentos Ortopédicos/estatística & dados numéricos , Complicações Pós-OperatóriasRESUMO
OBJECTIVE: To explore the mechanism of acupuncture in regulating chronic inflammation through dopamine in rats with chronic obstructive pulmonary disease (COPD). METHODS: 32 SD rats were randomly divided into control, model, sham acupuncture and acupuncture groups (n=8) . COPD condition was induced by eight-week exposure to cigarette smoking and lipopolysaccharides (LPS) of the rats, except for those in the control group. From the beginning of the 7th week, the acupuncture group received bilateral electroacupuncture on the Zusanli (ST-36), while the sham acupuncture group received bilateral electroacupuncture on the non-points, 30 min/time, 1/day, for 2 weeks prior to exposure to cigarette smoking. Post treatment changes in plasma dopamine and inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-8 (IL-8)], lung function [total lung capacity (TLC), functional residual capacity (FRC), the 50 µs forced expiratory volume (FEV) vs. forced vital capacity (FVC)( FEV50/FVC), the 100 µs FEV vs. FVC (FEV100/FVC), total airway resistance (RL), lung dynamic compliance (Cdyn)], and the ratio of total alveolus area to tissue area (A/t) and cell counts in the alveolar lavage fluid (BALF) were measured. Pearson correlations between plasma dopamine and the above indicators were calculated. RESULTS: Acupuncture increased plasma dopamine and improved the inflammatory factors, lung function, A/t and BALF cell counts. Compared with the model rats, the rats that received acupuncture had higher levels of TNF-α and IL-1ß, A/t and BALF cell counts, and lung function (FEV50/FVC, FEV100/FVC, RL, Cdyn) (P<0.05). The effects of acupuncture were superior on the ST-36 points compared with the non-points. Significant correlations between lung function (FRC, RL, Cdyn) and inflammatory factors (TNF-α, IL-1ß, IL-6, IL-8) were found (P<0.001) . TLC was correlated with IL-8, IL-1ß and A/t (P<0.05). Plasma dopamine was correlated with FRC, TLC, FEV50/FVC, FEV100/FVC (P<0.05). CONCLUSION: Acupuncture can alleviate inflammation, improve lung function and raise plasma dopamine level in COPD rats, and the effect of acupuncture on lung function may be related to reducing inflammatory factors and increasing dopamine level.
Assuntos
Terapia por Acupuntura , Dopamina/sangue , Pulmão/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/terapia , Pontos de Acupuntura , Animais , Eletroacupuntura , Interleucinas/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismoRESUMO
A series of new coumarin-dithiocarbamate hybrids were designed and synthesized as multitarget agents for the treatment of Alzheimer's disease. Most of them showed potent and clearly selective inhibition towards AChE and MAO-B. Among these compounds, compound 8f demonstrated the most potent inhibition to AChE with IC50 values of 0.0068⯵M and 0.0089⯵M for eeAChE and hAChE, respectively. Compound 8g was identified as the most potent inhibitor to hMAO-B, and it is also a good and balanced inhibitor to both hAChE and hMAO-B (0.114⯵M for hAChE; 0.101⯵M for hMAO-B). Kinetic and molecular modeling studies revealed that 8g was a dual binding site inhibitor for AChE and a competitive inhibitor for MAO-B. Further studies indicated that 8g could penetrate the BBB and exhibit no toxicity on SH-SY5Y neuroblastoma cells. More importantly, 8g did not display any acute toxicity in mice at doses up to 2500â¯mg/kg and could reverse the cognitive dysfunction of scopolamine-induced AD mice. Overall, these results highlighted 8g as a potential multitarget agent for AD treatment and offered a starting point for design of new multitarget AChE/MAO-B inhibitors based on dithiocarbamate scaffold.
Assuntos
Inibidores da Colinesterase/química , Inibidores da Colinesterase/farmacologia , Cumarínicos/química , Cumarínicos/farmacologia , Inibidores da Monoaminoxidase/química , Inibidores da Monoaminoxidase/farmacologia , Tiocarbamatos/química , Tiocarbamatos/farmacologia , Acetilcolinesterase/metabolismo , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/enzimologia , Animais , Linhagem Celular , Inibidores da Colinesterase/toxicidade , Cumarínicos/toxicidade , Desenho de Fármacos , Feminino , Humanos , Masculino , Camundongos , Simulação de Acoplamento Molecular , Monoaminoxidase/metabolismo , Inibidores da Monoaminoxidase/toxicidade , Tiocarbamatos/toxicidadeRESUMO
Novel hybrids with MAO and Aß (1-42) self-aggregation inhibitory activities were designed and synthesized with the employment of indazole moiety and resveratrol. The biological screening results indicated that most compounds displayed potent inhibitory activity for Aß (1-42) self-aggregation, and obvious selective inhibition to MAO-B. Among these compounds, compound 6e was the most potent inhibitor not only for hMAO-B (IC50â¯=â¯1.14⯵M) but also for Aß (1-42) self-aggregation (58.9% at 20⯵M). Molecular modeling and kinetic studies revealed that compound 6e was a competitive MAO-B inhibitor, which can occupy the active site of MAO-B, and interact with Aß (1-42) via π-π and cation-π stacking interactions. In addition, compound 6e had no toxicity on PC12 cells and could cross the BBB. Collectively, all these results suggested that compound 6e might be a promising multi-target lead compound worthy of further investigation.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Desenho de Fármacos , Indazóis/química , Inibidores da Monoaminoxidase/química , Fragmentos de Peptídeos/antagonistas & inibidores , Multimerização Proteica/efeitos dos fármacos , Resveratrol/análogos & derivados , Animais , Domínio Catalítico , Sobrevivência Celular/efeitos dos fármacos , Curcumina/farmacologia , Humanos , Indanos/farmacologia , Indazóis/síntese química , Indazóis/toxicidade , Iproniazida/farmacologia , Cinética , Simulação de Acoplamento Molecular , Monoaminoxidase/química , Inibidores da Monoaminoxidase/síntese química , Inibidores da Monoaminoxidase/toxicidade , Ratos , Resveratrol/síntese química , Resveratrol/toxicidadeRESUMO
A series of novel alkyl amine-substituted icariside II (ICA II) derivatives were synthesized by Mannich reactions at the 6-C position (compounds 4aâ»d) and changing the carbon chain length at the 7-OH position (compounds 7aâ»h), and their in vitro antitumor activity towards human breast cancer lines (MCF-7 and MDA-MB-231) and human hepatoma cell lines (HepG2 and HCCLM3-LUC) were evaluated by the MTT assay. Compared with ICA II, most of the twelve derivatives showed good micromole level activity and a preliminary structure-activity relationship (SAR) for the anticancer activity was obtained. Compound 7g showed the most potent inhibitory activity for the four cancer cell lines (13.28 µM for HCCLM3-LUC, 3.96 µM for HepG2, 2.44 µM for MCF-7 and 4.21 µM for MDA-MB-231), which was 2.94, 5.54, 12.56 and 7.72-fold stronger than that of ICA II. The preliminary SAR showed that the introduction of a alkyl amine substituent at 6-C was not favorable for the anticancer activity, while most of the 7-O-alkylamino derivatives exhibited good antitumor activity and the anticancer activity 7-O-alkylamino derivatives were influenced by the alkyl chain length and the different terminal amine substituents. Furthermore, the effects of compound 7g on apoptosis and cell cycle of MCF-7 cells were further investigated, which showed that compound 7g triggered apoptosis and arrested the cell cycle at the G0/G1 phase in MCF-7 cells. Our findings indicate that compound 7g may be a promising anticancer drug candidate lead.
Assuntos
Aminas , Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Flavonoides/síntese química , Flavonoides/farmacologia , Aminas/química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Flavonoides/química , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
OBJECTIVE: To investigate the effects and the underlying mechanisms of ShenFu Injection on paclitaxel-induced peripheral neuropathy. METHODS: Twenty-eight adult male Wister rats were randomized into 4 groups (n=7) : control group, paclitaxel group, paclitaxel combined with low or high dose of ShenFu Injection groups. Rats were intraperitoneally injected with paclitaxel 8 mg/kg every 4 d for a total of 4 doses except control group. From Day 1 of the experiment (injection),low dose (4 mL/kg) and high dose (8 mL/kg) of Shenfu Injection were intraperitoneally injected daily in the combination groups for a total of 21 d respectively,while normal saline (NS) was injected in control group in the same way instead. Mechanical withdraw threshold (MWT) and thermal withdraw latency (TWL) of rats' hind paw were measured before (0 d) and after the first injection (6 d,14 d). The level of nerve growth factor (NGF) in the serum was measured at 22 d before the euthanasia,and the ultrastructure of the sciatic nerve was observed with transmission electron microscope. RESULTS: The MWT and TWL of 14 d in paclitaxel group significantly increased compared with those of 0 d and control group ( P<0.05). The combination of paclitaxel with ShenFu Injection,especially the high dose ( P<0.05),significantly reduced the MWT and TWL when compared to paclitaxel group at 14 d. Compared with simultaneous control group,there was no remarkably increased MWT and TWL in the low and high dose of ShenFu Injection (P>0.05) . Compared with control group,the serum NGF level significantly decreased ( P<0.05) in paclitaxel group,while the serum NGF level in low and high dose of ShenFu Injection groups were higher than paclitaxel group,particularly in the high dose group ( P<0.05). When compared to control group,the sciatic nerve fiber structure in the paclitaxel group was generally damaged,including myelin sheath swelling,fragmentation and vacuolization,endoplasmic reticulum swelling and matrix structure disorder in Schwann cells. The structural damages were mitigated in the low dose and high dose groups,especially the latter one,when compared to the paclitaxel group. CONCLUSION: Shenfu Injection can reduce the peripheral neurotoxicity of paclitaxel by promoting the expression of NGF in serum.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Paclitaxel/efeitos adversos , Nervo Isquiático/efeitos dos fármacos , Animais , Masculino , Neurotoxinas/efeitos adversos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/ultraestruturaRESUMO
OBJECTIVE: To explore the curative effect of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKIs) combined with Traditional Chinese Medicine (TCM) versus single EGFR-TKIs for Advanced non-small-cell lung cancer (NSCLC). METHODS: A total of 59 NSCLC patients with EGFR mutation were divided (2:1) into treatment group and control group. Patients in treatment group (39 cases) take EGFR-TKIs plus TCM and control group (20 cases) take EGFR-TKIs. Analysis the progression-free survival (PFS), disease control rate (DCR) and treatment-related adverse events of two groups. RESULTS: The DCR of the treatment group and control group was 94.1% and 84.2% respectively (P=0.24). In the total population, PFS was 12.1 months in treatment group and 9.1 months in control group [hazard ratio (HR) 0.46; 95%CI 0.23-0.9; P=0.025]. Among patients with exon 19 deletion (19-del), PFS between treatment group and control group was 10.5 months and 9.5 months respectively (P=0.17). For patients with exon Leu858Arg point mutation (L858R), PFS was significantly longer with treatment group than withcontrol group (median 13.2 months vs. 7.8 months; HR 0.32, 95%CI 0.10-0.97; P=0.046). Grade 3-4 treatment-related adverse events were less common withtreatment-group (8.33 %) than control group (15.00%) (P=0.65). CONCLUSION: For NSCLC patients with EGFR mutation, EGFR-TKIs combined with TCM has a certain effect to prolong PFS, especially for the patients with L858R.