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1.
Mitochondrial DNA B Resour ; 6(11): 3134-3136, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34660891

RESUMO

We report complete chloroplast genome (plastome) sequences of Stylidium debile (150,105 bp) and Stylidium petiolare (150,998 bp). Both plastomes had the typical quadripartite structure, with large single-copy (LSC) and small single-copy (SSC) regions separated by two inverted repeat (IR) regions. Both plastomes have lost the rps19 and ycf15 CDS genes, and had infA-like, rps22-like, and rps7-like pseudogenes. Moreover, IR regions were expanded by having trnH GUG tRNA and the rps22-like pseudogene. Plastome phylogenomic analyses showed that the two Stylidium species formed a monophyletic clade (BS = 100), sister to the Argophyllaceae (BS = 86/83). Sequence differences between the two Stylidium plastomes were 5011 sites, including 2166 variable sites and 2845 indels, with the petA-psbJ spacer the most variable region, followed by the trnK UUU-matK intron and trnG UUG-rps16 spacer.

3.
Adv Rheumatol ; 61(1): 59, 2021 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-34538280

RESUMO

OBJECTIVE: To more precisely estimate the association between the tumor necrosis factor ligand superfamily member 4 (TNFSF4) gene polymorphisms and systemic lupus erythematosus (SLE) susceptibility, we performed a meta-analysis on the association of the following single nucleotide polymorphisms (SNPs) of TNFSF4 with SLE: rs1234315, rs844648, rs2205960, rs704840, rs844644, rs10489265. METHODS: A literature-based search was conducted using PubMed, MEDLINE, Embase, Web of Science databases, and Cochrane Library databases to identify all relevant studies. And the association of TNFSF4 gene polymorphisms and SLE susceptibility was evaluated by pooled odds ratio (OR) with 95% confidence interval (CI). RESULTS: The meta-analysis produced overall OR of 1.42 (95% CI 1.36-1.49, P < 0.00001), 1.41 (95% CI 1.36-1.46, P < 0.00001) and 1.34 (95% CI 1.26-1.42, P < 0.00001) for the rs2205960, rs1234315 and rs704840 polymorphisms respectively, confirming these three SNPs confer a significant risk for the development of SLE. On the other hand, the meta-analysis produced overall OR of 0.92 (95% CI 0.70-1.21, P = 0.54) for the rs844644 polymorphism, suggesting no significant association. And no association was also found between either rs844648 1.11 (OR 1.11, 95% CI 0.86-1.43, P = 0.41) or rs10489265 (OR 1.17, 95% CI 0.94-1.47, P = 0.17) polymorphism with SLE susceptibility, respectively. CONCLUSIONS: Our meta-analysis demonstrated that the TNFSF4 rs2205960, rs1234315 and rs844840 SNPs was significantly associated with an increased risk of SLE.

4.
J Control Release ; 337: 557-570, 2021 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-34371056

RESUMO

Current clinical agents for autoimmunity disorders treatment often cause substantial adverse effects and safety concerns, owing to non-specific immune modulation. Due to the prominent contribution of effector T cells in pathogenesis of rheumatoid arthritis (RA) and inflammatory bowel disease (IBD), and preferential location of co-stimulatory receptor-ligand pair OX40-OX40L at the inflamed sites, selectively targeting autoaggressive T cells by blockade OX40-OX40L, might represent an alternative strategy. Herein, we developed a new strategy to antagonize OX40-OX40L interaction by engineering a cell membrane derived nanovesicles (NVs) expressing OX40 receptors (OX40 NVs), and explored their potential for autoimmune disorders therapy. OX40 NVs showed specific binding capability to inflamed HUVECs in vitro, it also possessed distinct arthritic-targeting capacity in RA inflamed joints, and preferential accumulation in IBD inflamed colon. OX40 NVs efficiently suppressed the progression of both RA and IBD diseases through reducing CD4+OX40+ T cells population, and proinflammatory cytokines (i.e., TNF-α and IL-1ß), while reinforcing Tregs immune-suppressive effect, with superior therapeutic efficacy than anti-OX40L. Additionally, dexamethasone (DEX) loading can further enhance the potential of OX40 NVs for RA treatment. Owing to their preferential localization to inflamed sites, and potent immune-suppression ability, targeting OX40-OX40L blockade by OX40 NVs for autoimmune therapy is highly promising.

5.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(4): 539-542, 2021 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-34323027

RESUMO

Pharmaceutical sciences and pharmaceutical materials are a discipline that studies drug dosage forms, formulation technology, and drug release patterns and mechanisms. This article briefly discusses the fundamental research and application of pharmaceutical sciences/pharmaceutical materials, summarizes the hot research topics of pharmaceutical sciences/pharmaceutical materials in China over the past 20 years, and analyzes the international status, achievements, and strengths and weaknesses of research in pharmaceutical sciences/pharmaceutical materials in China. Suggestions were made for frontier research directions that should be prioritized in the field of pharmaceutical sciences and pharmaceutical materials in the next decade in China, and potential breakthrough that may be achieved was discussed, which may provide the basis and references for future work in the relevant areas.


Assuntos
Preparações Farmacêuticas , China , Previsões
6.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(4): 543-547, 2021 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-34323028

RESUMO

The transformersome is a new kind of lipoid drug delivery carrier. It has a number of excellent properties, including deformability, pressure permeability, and amphiphilicity. It has been widely used in the field of percutaneous and oral administration of medication. However, due to factors concerning its formulation, the stability and effectiveness of intravenous injection and other systemic routes of administration of transfersomes should be carefully examined. As an alternative, the formulation can be enhanced or improved in order to better exploit the strengths and avoid the weaknesses. Because of its deformability, transfersome may have distinctive potential strengths in the penetration of physiological barriers, for example, the blood-brain barrier, and in the research and development of transdermal immunization vaccines. This review has summarized five aspects of transfersomes, including the main properties, the formulation and process influencing factors, evaluation methods, main administration routes, and problems. Herein, we have given some examples and analysis, summarized the research achievements and assessd prospects for future development.


Assuntos
Sistemas de Liberação de Medicamentos , Lipossomos , Administração Cutânea , Portadores de Fármacos , Pesquisa , Pele
7.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(4): 630-636, 2021 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-34323042

RESUMO

Objective: To prepare encapsulated clopidogrel bisulfate (CLP) liposomes so as to deal with the poor water solubility of CLP, and to provide the experimental basis for the development of CLP formulations for intravascular injection. Methods: CLP-loaded liposomes were prepared using thin film hydration/sonication method and pH gradient active drug loading technology. Then, the morphology, particle size, encapsulation efficiency, drug loading capacity, Zeta potentials and in vitro release behavior were characterized. Bilateral renal arteries of Sprague-Dawley (SD) rats were clamped with micro-artery clamps to establish the model of renal ischemia-reperfusion injury (IRI) in male SD rats. The study aimed to preliminarily investigate the therapeutic effect of CLP-loaded liposome pretreatment on renal IRI in rats. Results: It was found that the optimal formulation and preparation technology of CLP liposomes were as follows: the CLP-to-phospholipid weight ratio of 1∶10, phospholipid-to-cholesterol ratio of 6∶1, octadecylamine-to-CLP ratio of 1.2∶1, PEG 400-to-CLP ratio of 1∶1, and incubation at 50 ℃ for 40 min. Then, following ultrasonication of 100 W efficiency at 5-second intervals for 20 times, CLP loading was conducted using 5 mL of 0.1 mol/L citric acid buffer at pH 3.0. Liposome samples were prepared with the film dispersion method, and the pH value was adjusted to 7.5 through pH gradient active drug loading technology. The CLP-loaded liposomes obtained in this way had a rounded shape, good dispersity, an average particle size of (134.13±2.60) nm, polydispersity index (PDI) of 0.25±0.02, and a Zeta potential of (2.12±0.23) mV. The encapsulation efficiency was found to be (98.66±0.14)%, and the drug loading capacity was (7.47±0.01)%. The in vitro release results showed that 66.24% of CLP was released cumulatively within 72 h. Preliminary efficacy experiments showed that animals pretreated with CLP-loaded liposomes had lower serum levels of blood urea nitrogen and creatinine compared to the levels of IRI model rats without any pretreatment. Conclusion: CLP-loaded liposomes were successfully prepared, which might provide the experimental foundation for the future development of CLP formulations for injection.


Assuntos
Lipossomos , Animais , Clopidogrel , Masculino , Tamanho da Partícula , Ratos , Ratos Sprague-Dawley , Solubilidade
8.
Biomaterials ; 275: 120902, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34087588

RESUMO

Activated fibroblasts are critical contributors to renal interstitial fibrosis thus becoming the cellular target for fibrosis treatment. Previously, microRNA 29 b (miR-29 b) is shown to be down-regulated in various animal models of renal fibrosis. Herein, we describe a facile strategy to achieve localized and sustained delivery of therapeutic microRNA to the kidney via a host-guest supramolecular hydrogel. Specifically, cationic bovine serum albumin is used to complex with miR-29 b to afford nanocomplexes (cBSA/miR-29 b), which is proven to specifically inhibit fibroblast activation in a dose-dependent manner in vitro. Following unilateral ureteral obstruction in mice, a single injection of the hydrogel loaded with cBSA/miR-29 b in vivo, significantly down-regulated proteins and genes related to fibrosis for up to 21 days without affecting the normal liver or kidney functions. Overall, the localized delivery of cBSA/miR-29 b via a host-guest supramolecular hydrogel represents a safe and effective intervention strategy to delay and reverse the progression of interstitial renal fibrosis.


Assuntos
Hidrogéis , Nefropatias , MicroRNAs , Obstrução Ureteral , Animais , Preparações de Ação Retardada , Fibrose , Rim/patologia , Nefropatias/patologia , Nefropatias/terapia , Camundongos , Camundongos Endogâmicos C57BL , MicroRNAs/uso terapêutico , Obstrução Ureteral/patologia , Obstrução Ureteral/terapia
9.
Sci Total Environ ; 789: 147915, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34049145

RESUMO

Tenebrio obscurus (Coleoptera: Tenebrionidae) larvae are capable of biodegrading polystyrene (PS) but their capacity for polyethylene (PE) degradation and pattern of depolymerization remains unknown. This study fed the larvae of T. obscurus and Tenebrio molitor, which have PE degrading capacity, two commercial low-density PE (LDPE) foams i.e., PE-1 and PE-2, with respective number-average molecular weights (Mn) of 28.9 and 27.3 kDa and weight-average molecular weights (Mw) of 342.0 and 264.1 kDa, over a 36-day period at ambient temperature. The Mw of residual PE in frass (excrement) of T. obscurus, fed with PE-1 and PE-2, decreased by 45.4 ± 0.4% and 34.8 ± 0.3%, respectively, while the respective decrease in frass of T. molitor was 43.3 ± 0.5% and 31.7 ± 0.5%. Data analysis showed that low molecular weight PE (<5.0 kDa) was rapidly digested while longer chain portions (>10.0 kDa) were broken down or cleaved, indicating a broad depolymerization pattern. Mass balance analysis indicated nearly 40% of ingested LDPE was digested to CO2. Antibiotic suppression of gut microbes in T. molitor and T. obscurus larvae with gentamicin obviously reduced their gut microbes on day 15 but did not stop depolymerization because the Mn, Mw and size- average molecular weight (Mz) decreased. This confirmed that LDPE biodegradation in T. obscurus was independent of gut microbes as observed during previous PS degradation in T. molitor, suggesting that the intestinal digestive system could perform LDPE depolymerization. High-throughput sequencing revealed significant shifts in the gut microbial community during bran-fed and unfed conditions in response to LDPE feeding in both Tenebrio species. The respective predominant gut genera of Spiroplasma sp. and Enterococcus sp. were observed in LDPE-fed T. molitor and T. obscurus larvae.


Assuntos
Besouros , Microbioma Gastrointestinal , Tenebrio , Animais , Larva , Polietileno
10.
Acta Pharm Sin B ; 11(3): 835-847, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33777685

RESUMO

Localized delivery, comparing to systemic drug administration, offers a unique alternative to enhance efficacy, lower dosage, and minimize systemic tissue toxicity by releasing therapeutics locally and specifically to the site of interests. Herein, a localized drug delivery platform ("plum‒pudding" structure) with controlled release and long-acting features is developed through an injectable hydrogel ("pudding") crosslinked via self-assembled triblock polymeric micelles ("plum") to help reduce renal interstitial fibrosis. This strategy achieves controlled and prolonged release of model therapeutics in the kidney for up to three weeks in mice. Following a single injection, local treatments containing either anti-inflammatory small molecule celastrol or anti-TGFß antibody effectively minimize inflammation while alleviating fibrosis via inhibiting NF-κB signaling pathway or neutralizing TGF-ß1 locally. Importantly, the micelle-hydrogel hybrid based localized therapy shows enhanced efficacy without local or systemic toxicity, which may represent a clinically relevant delivery platform in the management of renal interstitial fibrosis.

11.
Acta Pharmacol Sin ; 42(7): 1040-1054, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33772141

RESUMO

Nano drug-delivery systems (DDS) may significantly improve efficiency and reduce toxicity of loaded drugs, but a few nano-DDS are highly successful in clinical use. Unprotected nanoparticles in blood flow are often quickly cleared, which could limit their circulation time and drug delivery efficiency. Elongating their blood circulation time may improve their delivery efficiency or grant them new therapeutic possibilities. Erythrocytes are abundant endogenous cells in blood and are continuously renewed, with a long life span of 100-120 days. Hence, loading nanoparticles on the surface of erythrocytes to protect the nanoparticles could be highly effective for enhancing their in vivo circulation time. One of the key questions here is how to properly attach nanoparticles on erythrocytes for different purposes and different types of nanoparticles to achieve ideal results. In this review, we describe various methods to attach nanoparticles and drugs to the erythrocyte surface, and discuss the key factors that influence the stability and circulation properties of the erythrocytes-based delivery system in vivo. These data show that using erythrocytes as a host for nanoparticles possesses great potential for further development.

12.
Mitochondrial DNA B Resour ; 6(1): 293-294, 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33553648

RESUMO

Aletris megalantha F. T. Wang & Tang is an herbal plant species endemic to Yunnan Province of China. Its complete plastid genome sequence was 154,704 bp in length, with a large single-copy (LSC) region of 83,265 bp, a small single-copy (SSC) region of 18,127 bp, and a pair of inverted repeat regions (IRs) of 26,656 bp. The whole plastid genome encoded 132 genes, including 85 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The overall GC content of A. megalantha plastid genome was 37.4%. Maximum likelihood phylogenetic analysis based on 14 taxa indicated that A. megalantha is evolutionarily close to A. spicata.

13.
Adv Rheumatol ; 61: 59, 2021. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1339072

RESUMO

Abstract Objective: To more precisely estimate the association between the tumor necrosis factor ligand superfamily member 4 (TNFSF4) gene polymorphisms and systemic lupus erythematosus (SLE) susceptibility, we performed a metaanalysis on the association of the following single nucleotide polymorphisms (SNPs) of TNFSF4 with SLE: rs1234315, rs844648, rs2205960, rs704840, rs844644, rs10489265. Methods: A literature-based search was conducted using PubMed, MEDLINE, Embase, Web of Science databases, and Cochrane Library databases to identify all relevant studies. And the association of TNFSF4 gene polymorphisms and SLE susceptibility was evaluated by pooled odds ratio (OR) with 95% confidence interval (CI). Results: The meta-analysis produced overall OR of 1.42 (95% CI 1.36-1.49, P < 0.00001), 1.41 (95% CI 1.36-1.46, P < 0.00001) and 1.34 (95% CI 1.26-1.42, P < 0.00001) for the rs2205960, rs1234315 and rs704840 polymorphisms respectively, confirming these three SNPs confer a significant risk for the development of SLE. On the other hand, the meta-analysis produced overall OR of 0.92 (95% CI 0.70-1.21, P = 0.54) for the rs844644 polymorphism, suggesting no significant association. And no association was also found between either rs844648 1.11 (OR 1.11, 95% CI 0.86-1.43, P = 0.41) or rs10489265 (OR 1.17,95% CI 0.94-1.47, P = 0.17) polymorphism with SLE susceptibility, respectively. Conclusions: Our meta-analysis demonstrated that the TNFSF4 rs2205960, rs1234315 and rs844840 SNPs was significantly associated with an increased risk of SLE.

14.
PhytoKeys ; 157: 27-41, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32934446

RESUMO

The former genus Bournea is endemic to China, including two species, has been under consideration for incorporation into the expanded genus Oreocharis s.l. in Gesneriaceae. The phylogenetic tree inferred from two DNA sequences (trnL-F and ITS) showed that this genus is deeply nested into Oreocharis s.l. However, the new tree from seven ones (atpB-rbcL, ndhH-rps15-ycf1, rpl132, trnC-trnD, trnL-F, trnT-trnL of chloroplast DNA and ITS regions) revealed that Bournea is the sister group of other of Oreocharis s.l. Furthermore, Bournea is morphologically different from other Oreocharis based on existing data. We suggest keeping Bournea as an independent genus in Gesneriaceae.

15.
Front Plant Sci ; 11: 793, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32636856

RESUMO

The liquorice genus, Glycyrrhiza L. (Leguminosae), is a medicinal herb with great economic importance and an intriguing intercontinental disjunct distribution in Eurasia, North Africa, the Americas, and Australia. Glycyrrhiza, along with Glycyrrhizopsis Boiss. and Meristotropis Fisch. & C.A.Mey., comprise Glycyrrhiza s.l. Here we reconstructed the phylogenetic relationships and biogeographic history in Glycyrrhiza s.l. using sequence data of whole chloroplast genomes. We found that Glycyrrhiza s.l. is sister to the tribe Wisterieae and is divided into four main clades. Clade I, corresponds to Glycyrrhizopsis and is sister to Glycyrrhiza sensu Meng. Meristotropis is embedded within Glycyrrhiza sensu Meng, and these two genera together form Clades II-IV. Based on biogeographic analyses and divergence time dating, Glycyrrhiza s.l. originated during the late Eocene and its most recent common ancestor (MRCA) was distributed in the interior of Eurasia and the circum-Mediterranean region. A vicariance event, which was possibly a response to the uplifting of the Turkish-Iranian Plateau, may have driven the divergence between Glycyrrhiza sensu Meng and Glycyrrhizopsis in the Middle Miocene. The third and fourth main uplift events of the Qinghai-Tibetan Plateau may have led to rapid evolutionary diversification within Glycyrrhiza sensu Meng. Subsequently, the MRCA of Clade II might have migrated to North America (G. lepidota) via the Bering land bridge during the early Pliocene, and reached temperate South America (G. astragalina) by long-distance dispersal (LDD). Within Clade III, the ancestor of G. acanthocarpa arrived at southern Australia through LDD after the late Pliocene, whereas all other species (the SPEY clade) migrated to the interior of Eurasia and the Mediterranean region in the early Pleistocene. The MRCA of Clade IV was restricted in the interior of Eurasia, but its descendants have become widespread in temperate regions of the Old World Northern Hemisphere during the last million years.

16.
Shanghai Kou Qiang Yi Xue ; 29(2): 217-220, 2020 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-32626889

RESUMO

PURPOSE: To investigate the clinical effects of oral implant restoration in patients with dentition defects and the its impact on tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels in gingival crevicular fluid. METHODS: Eighty-four patients with dentition defects from May 2017 to August 1919 in People's Hospital of Shanghai Pudong District were enrolled and randomly divided into control group (n=42) and experimental group (n=42). Patients in the control group were repaired by routine methods,while those in the experimental group were treated with oral implant restoration. The effect of restoration was evaluated 6 months after treatment. The levels of TNF-α, IL-6 in the gingival crevicular fluid and dental function were compared between the 2 groups. The data were analyzed using SPSS 18.0 software package. RESULTS: The levels of TNF-α and IL-6 in the experimental group and the control group after treatment were significantly higher than those before treatment (P<0.05). The levels of TNF-α and IL-6 in the experimental group were significantly lower than those in the control group 6 months after treatment (P<0.05). The scores of dental function in the experimental group and the control group were significantly higher than those before treatment (P<0.05). The scores of retention, speech, chewing and aesthetics of the experimental group 6 months after treatment were significantly higher than the control group (P<0.05). The incidence of infection, pricking, post and core loosing and teeth missing in the experimental group was significantly lower than that of the control group (P<0.05). CONCLUSIONS: In the treatment of patients with dentition defects, implant restoration has little effect on the levels of TNF-α and IL-6 in gingival crevicular fluid, which is helpful to improve dental function and reduce the incidence of postoperative complications. Therefore, it is worthwhile to be popularized in clinical application.


Assuntos
Implantes Dentários , Líquido do Sulco Gengival , China , Dentição , Estética Dentária , Humanos , Interleucina-6 , Índice Periodontal , Fator de Necrose Tumoral alfa
17.
J Mater Chem B ; 8(10): 2139-2147, 2020 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-32090232

RESUMO

Due to the critical role of CD44 in mediating cell adhesion and migration, CD44-targeted drug delivery via hyaluronan has been extensively explored. Herein, cationic bovine serum albumin nanoparticles were assembled with hyaluronan (HA) of various molecular weights via simple electrostatic interaction to afford hierarchical nanoparticles (HNPs) with various size distributions and structures. Next, HNPs obtained using 49 kDa HA have been used to encapsulate paclitaxel (PTX-HNPs), which demonstrated selective lung accumulation due to both size effect and CD44-mediated targetability. Biodistribution studies showed that HNPs enhanced the lung specific accumulation of HNPs in the C57BL/6 mice melanoma lung metastasis model. In the antitumor studies, compared with the Taxol or bovine serum albumin nanoparticle (NP) groups, PTX-HNPs significantly inhibited B16F10 lung metastasis at a relatively low dose. Additionally, cell migration and invasion experiments in vitro further confirmed that PTX-HNPs significantly inhibited the migration of B16F10 cells compared to Taxol or paclitaxel-loaded NP groups. Overall, our results suggest that PTX-HNPs represent a highly promising strategy for the treatment of lung metastatic melanoma.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Ácido Hialurônico/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Melanoma/tratamento farmacológico , Nanopartículas/química , Paclitaxel/farmacologia , Soroalbumina Bovina/química , Animais , Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/química , Apoptose/efeitos dos fármacos , Cátions/síntese química , Cátions/química , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Ácido Hialurônico/química , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/secundário , Melanoma/patologia , Melanoma/secundário , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/patologia , Neoplasias Experimentais/secundário , Paclitaxel/química , Tamanho da Partícula , Soroalbumina Bovina/síntese química , Propriedades de Superfície , Células Tumorais Cultivadas
18.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(1): 81-86, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31950794

RESUMO

Objective: To preliminarily investigate the differences of protein composition between immature dendritic cells (DC2.4) and their derived exosomes (DC-Exo) using a relatively rapid and sample-saving method based on nano-flow liquid chromatography tandem mass spectrometry (nanoLC-MS/MS). Methods: The supernatant of DC2.4 cells culture medium was collected and gradient centrifugation was applied to primarily extract and isolate DC-Exo; then sucrose density gradient ultracentrifugation was adopted to purify the DC-Exo. Bradford protein assay was used to determine the total protein content of the purified DC-Exo, and dynamic light scattering and transmission electron microscope were conducted to characterize the morphology and size distribution of the DC-Exo. Afterwards, protein samples including DC2.4 cells and DC-Exo were prepared by FASP enzymolysis method. Samples were performed nanoLC-MS/MS assay. The µLPickUp sample loading mode was used and only 1 µg of protein sample was required for each assay. The phase of Transport liquid and Micro A were both 0.05% trifluoroacetic acid-2% acetonitrile (ACN) aq. ( V/ V). Acclaim ® PepMap RSLC column was used to separate sample compositions and the gradient elute was adopted where the mobile phase consisted of (A) 0.1% formic acid (FA) and (B) 0.08% FA-80% ACN aq. ( V/ V) with flow rate of 0.3 µL/min. Positive APCI nanospray interface was used and "one-drive-ten" schema was set to collect primary information. The collected data was then searched and matched based on Uniport Mouse Fasta file as protein database in this case, and the re-annotated data was further sorted out and analyzed. Results: In the current study, relatively high yield of DC-Exo samples with sizes of 40-200 nm were obtained. The lyophilized protein samples prepared by FASP method could be loaded directly after redissolution, and only 1 µg of protein sample is required. The annotated results showed that DC2.4 cells contained 998 kinds of proteins, among which 227 were highly expressed and 535 were unique; while DC-Exo contained only 348 types of proteins, among which 18 were uniquely and highly expressed. There were 306 kinds of consensus proteins in both DC2.4 cells and DC-Exo, among them 7 kinds were highly expressed. Conclusion: The nanoLC-MS/MS method developed in this study only requires very small amount of protein samples, and it could primarily differentiate the protein compositions between DC2.4 cells and their derived exosomes rapidly.


Assuntos
Cromatografia Líquida , Células Dendríticas , Exossomos , Proteínas , Espectrometria de Massas em Tandem , Animais , Células Dendríticas/química , Exossomos/química , Camundongos , Proteínas/química
19.
J Mater Chem B ; 8(3): 578-579, 2020 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-31930251

RESUMO

Correction for 'A fast-dissolving microneedle array loaded with chitosan nanoparticles to evoke systemic immune responses in mice' by Zhilin Li et al., J. Mater. Chem. B, 2020, DOI: 10.1039/c9tb02061f.

20.
J Mater Chem B ; 8(2): 216-225, 2020 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-31803892

RESUMO

Microneedle (MN) arrays offer an alternative approach to hypodermic injection via syringe needles. In this work, polyvinylpyrrolidone (PVP)-based fast dissolving MN arrays were developed in which the needle tips were loaded with chitosan nanoparticles (NPs) for coencapsulation of a model antigen, ovalbumin (OVA), and an adjuvant, CpG oligodeoxynucleotides (CpG). After insertion into the skin, these MN arrays fully dissolved within 3 min to release antigen and adjuvant co-loaded NPs rapidly in the epidermal layer. Positively charged chitosan was proven to be an excellent carrier for negatively charged OVA and CpG, which formed nanocomplexes via simple electrostatic interactions and greatly enhanced the uptake efficiency of OVA in DC2.4 dendritic cells. Vaccination studies in mice further demonstrated that chitosan NPs effectively accumulated in peripheral lymph nodes, thus inducing greatly enhanced immune responses compared to those of free OVA. The antibody dose-response curve further demonstrated that MN immunization achieved comparable levels of immune responses as compared to conventional subcutaneous injections in a more convenient and less invasive way. Overall, a PVP-based fast dissolving MN array with chitosan NPs represents a promising and robust platform system for efficient transcutaneous vaccine delivery.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Imunização/métodos , Injeções Intradérmicas/métodos , Microinjeções/métodos , Oligodesoxirribonucleotídeos/farmacologia , Ovalbumina/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/farmacologia , Administração Cutânea , Animais , Antígenos/administração & dosagem , Antígenos/farmacologia , Quitosana/química , Feminino , Camundongos , Nanopartículas/química , Oligodesoxirribonucleotídeos/administração & dosagem , Ovalbumina/administração & dosagem
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