RESUMO
Objective: To introduce a new self-developed irrigation device(SID) that does not employ a sheath or an irrigation-suction system and evaluate to its efficiency in transcanal endoscopic ear surgery (TEES) for attic cholesteatoma. Methods: 38 patients who were subjected to TEES for attic cholesteatoma between October 2019 to June 2021 were included in this study, including 17 males and 21 females with an average age of (38.6±11.9) years. SID and underwater continuous drilling were used during operation. Width of endoscope and irrigation speed were measured when SID was applied. The operating time, surgical view and complications were compared between two groups. Results: The width of the endoscope was 3.5-4.6 mm in diameter and the irrigation speed was 20-40 ml/min when SID was used. SID cleaned the lens at the tip of the endoscope and created a clear field of view during TEES. The operation time was (86.6±18.1) min. The skin of the external ear canal was found injured during operation in 3 patients, but there were no complications such as necrosis of the flap, stenosis of external ear canal, sensorineural hearing loss, facial paralysis and cerebrospinal fluid leakage. Conclusions: SID is simple and enhances the efficacy of TEES, providing a new irrigation choice in TEES for attic cholesteatoma.
Assuntos
Colesteatoma da Orelha Média , Procedimentos Cirúrgicos Otológicos , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Colesteatoma da Orelha Média/cirurgia , Orelha Média/cirurgia , SucçãoAssuntos
Fígado Artificial , Trombocitopenia , Anticoagulantes/efeitos adversos , Arginina/análogos & derivados , Heparina/efeitos adversos , Humanos , Ácidos Pipecólicos/uso terapêutico , Terapia de Salvação , Sulfonamidas , Trombocitopenia/induzido quimicamente , Trombocitopenia/tratamento farmacológico , Resultado do TratamentoRESUMO
Objective: To evaluate the results of butterfly cartilage myringoplasty for anterior quadrant tympanic perforation under endoscope. Methods: Thirty-eight patients with anterior quadrant tympanic perforations who were subjected to endoscopic butterfly cartilage myringoplasty from April 2016 to October 2018 were included in this study, including 16 males and 22 females, with an average age of (34.5±14.2) years. The patients were reviewed retrospectively, and the pre-and post-operative pure tone audiometry (PTA) thresholds, pre-and post-operative air-bone gaps (ABG), post-operative graft success rates and complications were evaluated. SPSS 23.0 was used to analyze data. Results: Mean post-operative follow-up duration was (9.4±3.1) months (range 6-18 months). The graft survival rate was 94.7% (36/38) . The preoperative and postoperative mean PTA was (30.9±8.9) dB HL and (21.4±7.7) dB HL respectively. Preoperative and postoperative mean ABG was (18.4±6.3) dB and (10.8±6.0) dB respectively. There was significant difference between pre-and postoperative PTA and ABG (t=5.353 and 4.162, P<0.05 for both). A postoperative ABG reduction of (8.3±1.5) dB was reached. Two (4.7%) patients had postoperative myringitis, two (4.7%) had recurrent perforation, and one (2.4%) had lateral healing of transplanted tympanic membrane in the postoperative follow-ups. No intratympanic cholesteatoma was observed. Conclusions: Endoscopic butterfly inlay myringoplasty is a reliable, minimally invasive alternative method to repair anterior tympanic membrane perforations, with high closure rate and low risk of complications.
Assuntos
Miringoplastia , Perfuração da Membrana Timpânica , Adulto , Cartilagem , Endoscópios , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Membrana Timpânica , Perfuração da Membrana Timpânica/cirurgia , Adulto JovemRESUMO
Objective: To introduce a self-developed bone dust collector designed by the authors and evaluate its efficiency in mastoid obliteration following mastoidectomy. Methods: Consecutive patients, from April 2017 to March 2018, who prepared to receive mastoidectomy were randomly divided into two groups, and in each group the bone dust was harvested by self-developed bone dust collector or by conventional used method respectively in mastoidectomy. The amount of the harvested bone dust and the time consumed in the collecting procedure were compared between two groups. The infection of the bone dust after mastoid obliteration was also evaluated during follow up. Results: 33 patients were recruited in bone dust collector group, and 31 patients in conventional method group.There is no significance of difference between two groups in sex ratio, age and pneumatization of mastoid cells (P>0.05 for all). The median amount of bone dust harvested by bone dust collector was significantly larger than that collected by conventional method (1.8 g vs 1.1 g, P<0.05). The median time spent in bone dust collector group was significantly shorter than that spent in conventional method group (4 minutes vs 6 minutes, P<0.05). No bone dust infection was found in the follow-up in all patients. Conclusion: The present self-developed bone dust collector is a easy and useful apparatus which can significantly improve the efficiency of collecting bone dust in mastoidectomy.
Assuntos
Poeira , Processo Mastoide/cirurgia , Mastoidectomia/instrumentação , Manejo de Espécimes/instrumentação , Feminino , Humanos , MasculinoRESUMO
Graphitization occurs during the long-term service of a diamond-like carbon (DLC) modified artificial joint. Then, DLC wear debris, which are carbon particles with different sp2/sp3 ratios and sizes ranging from the nano- to micro-meter scale produced. In this paper, to promote the application of DLC coating for artificial joint modification, the cytotoxicity of DLC debris (nano-carbon particles, NCs) with different sp2/sp3 ratios was studied. The microstructure and physical characteristics of NCs with different sp2/sp3 ratios were investigated by Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), Transmission Electron Microscope (TEM) and Dynamic Light Scattering (DLS). Meanwhile, osteoblasts and macrophages were applied to characterize the cytotoxicity of the NCs. In vitro cytotoxicity assay results indicated that cells incubated with NCs of different sp2/sp3 ratios had greater osteogenic capacity, and these particles caused a weaker immune response in comparison with CoCrMo particles. Taken together, the results indicated that NCs with different sp2/sp3 ratios presented a good cytocompatibility than CoCrMo particles. But no significant differences were observed among NCs with different sp2/sp3 ratios. The better cytocompatibility of NCs is mainly attributable to their surface charge.
Assuntos
Carbono/toxicidade , Nanoestruturas/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Interleucina-6/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos , Microscopia Eletrônica de Transmissão , Modelos Teóricos , Nanoestruturas/química , Osteoblastos/efeitos dos fármacos , Espectroscopia Fotoeletrônica , Células RAW 264.7 , Fator de Necrose Tumoral alfa/metabolismoRESUMO
While a diamond-like carbon (DLC)-coated joint prosthesis represents the implant of choice for total hip replacement in patients, it also leads to concern due to the cytotoxicity of wear debris in the form of graphite nanoparticles (GNs), ultimately limiting its clinical use. In this study, the cytotoxicity of various GN doses was evaluated. Mouse macrophages and osteoblasts were incubated with GNs (<30 nm diameter), followed by evaluation of cytotoxicity by means of assessing inflammatory cytokines, results of alkaline phosphatase assays, and related signaling protein expression. Cytotoxicity evaluation showed that cell viability decreased in a dose-dependent manner (10-100 µg ml-1), and steeply declined at GNs concentrations greater than 30 µg ml-1. Noticeable cytotoxicity was observed as the GN dose exceeded this threshold due to upregulated receptor of activator of nuclear factor kB-ligand expression and downregulated osteoprotegerin expression. Meanwhile, activated macrophage morphology was observed as a result of the intense inflammatory response caused by the high doses of GNs (>30 µg ml-1), as observed by the increased release of TNF-α and IL-6. The results suggest that GNs had a significant dose-dependent cytotoxicity in vitro, with a lethal dose of 30 µg ml-1 leading to dramatic increases in cytotoxicity. Our GN cytotoxicity evaluation indicates a safe level for wear debris-related arthropathy and could propel the clinical application of DLC-coated total hip prostheses.
Assuntos
Materiais Revestidos Biocompatíveis/toxicidade , Grafite/toxicidade , Prótese Articular , Nanopartículas/toxicidade , Animais , Carbono/química , Células Cultivadas , Materiais Revestidos Biocompatíveis/administração & dosagem , Materiais Revestidos Biocompatíveis/química , Diamante/química , Relação Dose-Resposta a Droga , Grafite/administração & dosagem , Grafite/química , Humanos , Mediadores da Inflamação/metabolismo , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Teste de Materiais , Camundongos , Nanopartículas/administração & dosagem , Nanopartículas/química , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteogênese/efeitos dos fármacos , Falha de Prótese , Células RAW 264.7RESUMO
Diamond-like carbon (DLC) films are potential candidates for artificial joint surface modification in biomedical applications, and the influence of the structural features of DLC surfaces on cell functions has attracted attention in recent decades. Here, the biocompatibility of DLC films with different structures was investigated using macrophages, osteoblasts and fibroblasts. The results showed that DLC films with a low ratio of sp(2)/sp(3), which tend to have a structure similar to that of diamond, led to less inflammatory, excellent osteogenic and fibroblastic reactions, with higher cell viability, better morphology, lower release of TNF-α (tumor necrosis factor-α) and IL-6 (interleukin-6), and higher release of IL-10 (interleukin-10). The results also demonstrated that the high-density diamond structure (low ratio of sp(2)/sp(3)) of DLC films is beneficial for cell adhesion and growth because of better protein adsorption without electrostatic repulsion. These findings provide valuable insights into the mechanisms underlying inhibition of an inflammatory response and the promotion of osteoblastogenesis and fibrous propagation, and effectively build a system for evaluating the biocompatibility of DLC films.
Assuntos
Tecnologia Biomédica/métodos , Diamante/química , Diamante/farmacologia , Adsorção , Animais , Bovinos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Mediadores da Inflamação/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/ultraestrutura , Camundongos , Microscopia de Força Atômica , Microscopia de Fluorescência , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Soroalbumina Bovina/química , Análise Espectral RamanRESUMO
BACKGROUND: The patatin-like phospholipase 3 (PNPLA3) rs738409 gene polymorphism is an important genetic determinant of non-alcoholic fatty liver disease (NAFLD). However, the associations between liver fat and metabolic traits in rs738409 G allele carriers and the allelic influence on this association have not been fully studied. AIM: To investigate the influence of the PNPLA3 gene polymorphism on the association of liver fat with serum metabolic factors and carotid atherosclerosis. METHODS: Liver fat was measured by quantitative ultrasound in 4300 subjects in the Shanghai Changfeng community and analysed for its association with obesity and metabolic factors in individuals with the PNPLA3 CC, CG and GG genotypes. RESULTS: Non-alcoholic fatty liver disease occurred in 37.9% and 28.8% of the subjects with the GG and CC genotypes respectively (P < 0.001). Liver fat was significantly associated with body mass index, waist circumference, serum triglycerides, high-density lipoprotein cholesterol, fasting blood glucose and insulin in the PNPLA3 rs738409 G allele carriers (P < 0.001). Compared with the CC homozygotes, the GG homozygotes presented higher liver fat and liver fibrosis scores despite their better metabolic status (comparison of regression line slopes, P < 0.05). An increase in liver fat was accompanied by a significant increase in the average and maximum carotid intima-media thickness in subjects with the PNPLA3 CC genotype but not in those with the GG genotype. CONCLUSIONS: PNPLA3 rs738409 G allele carriers were found to be more susceptible to the metabolic-related hepatic steatosis, and developed NAFLD and liver fibrosis despite presenting relatively better metabolic statuses and lower risks for carotid atherosclerosis.
Assuntos
Lipase/genética , Proteínas de Membrana/genética , Hepatopatia Gordurosa não Alcoólica/genética , Obesidade/fisiopatologia , Idoso , Alelos , Glicemia , Índice de Massa Corporal , Pesos e Medidas Corporais , Espessura Intima-Media Carotídea , China , HDL-Colesterol/sangue , Feminino , Genótipo , Humanos , Fígado/patologia , Masculino , Pessoa de Meia-Idade , Fosfolipases , Polimorfismo Genético , Fatores de RiscoRESUMO
Epigenetic modifications of the genome, such as histone H2A variants, ensure appropriate gene activation or silencing during oogenesis and preimplantation embryo development. We examined global localization and expression of the histone H2A variants, including H2A.Bbd, H2A.Z and H2A.X, during mouse oogenesis and preimplantation embryo development. Immunocytochemistry with specific antibodies against various histone H2A variants showed their localization and changes during oogenesis and preimplantation development. H2A.Bbd and H2A.Z were almost absent from nuclei of growing oocytes (except 5-day oocyte), whereas H2A.X was deposited in nuclei throughout oogenesis and in preimplantation embryos. In germinal vesicle (GV) oocyte chromatin, H2A.Bbd was detected as a weak signal, whereas no fluorescent signal was detected in GV breakdown (GVBD) or metaphase II (MII) oocytes; H2A.Z showed intense signals in chromatin of GV, GVBD and MII oocytes. H2A. Bbd showed very weak signals in both pronucleus and 2-cell embryo nuclei, but intense signals were detected in nuclei from 4-cell embryo to blastula. The H2A.Z signal was absent from pronucleus to morula chromatin, whereas a fluorescent signal was detected in blastula nuclei. Our results suggest that histone H2A variants are probably involved in reprogramming of genomes during oocyte meiosis or after fertilization.
Assuntos
Blastocisto/metabolismo , Desenvolvimento Embrionário/genética , Expressão Gênica , Histonas/genética , Oogênese/genética , Animais , Feminino , Histonas/metabolismo , Imuno-Histoquímica , Meiose , Camundongos , Gravidez , Transporte ProteicoRESUMO
Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.
Assuntos
Ácido 3-Hidroxibutírico/química , Materiais Biocompatíveis/química , Caproatos/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Ácido 3-Hidroxibutírico/uso terapêutico , Materiais Biocompatíveis/uso terapêutico , Osso e Ossos/fisiologia , Caproatos/uso terapêutico , Cartilagem/fisiologia , Liofilização , Humanos , Músculo Liso/fisiologia , Regeneração , Propriedades de SuperfícieRESUMO
Development and selection of an ideal scaffold is of importance for tissue engineering. Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a biocompatible bioresorbable copolymer that belongs to the polyhydroxyalkanoate family. Because of its good biocompatibility, PHBHHx has been widely used as a cell scaffold for tissue engineering. This review focuses on the utilization of PHBHHx-based scaffolds in tissue engineering. Advances in the preparation, modification, and application of PHBHHx scaffolds are discussed.
Assuntos
Humanos , /química , Materiais Biocompatíveis/química , Caproatos/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , /uso terapêutico , Materiais Biocompatíveis/uso terapêutico , Osso e Ossos/fisiologia , Caproatos/uso terapêutico , Cartilagem/fisiologia , Liofilização , Músculo Liso/fisiologia , Regeneração , Propriedades de SuperfícieRESUMO
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Assuntos
Humanos , Diferenciação Celular/genética , Condrogênese/genética , Sangue Fetal/citologia , Células-Tronco Mesenquimais/citologia , Fatores de Transcrição SOX9/genética , Agrecanas/biossíntese , Western Blotting , Cartilagem/metabolismo , Proliferação de Células/genética , Condrócitos/metabolismo , Colágeno Tipo II/biossíntese , Citometria de Fluxo , Proteínas de Fluorescência Verde , Regulação da Expressão Gênica/fisiologia , Células Endoteliais da Veia Umbilical Humana/citologia , Imuno-Histoquímica , Imunofenotipagem , Cultura Primária de Células , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Engenharia Tecidual , TransfecçãoRESUMO
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Assuntos
Diferenciação Celular/genética , Condrogênese/genética , Sangue Fetal/citologia , Células-Tronco Mesenquimais/citologia , Fatores de Transcrição SOX9/genética , Agrecanas/biossíntese , Western Blotting , Cartilagem/metabolismo , Proliferação de Células/genética , Condrócitos/metabolismo , Colágeno Tipo II/biossíntese , Citometria de Fluxo , Regulação da Expressão Gênica/fisiologia , Proteínas de Fluorescência Verde , Células Endoteliais da Veia Umbilical Humana/citologia , Humanos , Imuno-Histoquímica , Imunofenotipagem , Cultura Primária de Células , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Engenharia Tecidual , TransfecçãoRESUMO
MicroRNAs (miRNAs) are potent regulators of gene transcription and posttranscriptional processes. The majority of miRNAs are localized within intronic regions of protein-coding genes (host genes) and have diverse functions in regulating important cellular processes in animals. To date, few plant intronic miRNAs have been studied functionally. Here we report a comprehensive computational analysis to characterize intronic miRNAs in rice and Arabidopsis. RT-PCR analysis confirmed that the identified intronic miRNAs were derived from the real introns of host genes. Interestingly, 13 intronic miRNAs in rice and two in Arabidopsis were located within seven clusters, of which four polycistronic clusters contain miRNAs derived from different families, suggesting that these clustered intronic miRNAs might be involved in extremely complex regulation in rice. Length analysis of miRNA-carrying introns, promoter prediction and qRT-PCR analysis results indicated that intronic miRNAs are coexpressed with their host genes. Expression pattern analysis demonstrated that host genes had a very broad expression spectrum in different stages of development, suggesting the intronic miRNAs might play an important role in plant development. This comparative genomics analysis of intronic miRNAs in rice and Arabidopsis provides new insight into the functions and regulatory mechanisms of intronic miRNAs in monocots and dicots.
Assuntos
Arabidopsis/genética , Genoma de Planta/genética , Íntrons/genética , MicroRNAs/genética , Oryza/genética , Arabidopsis/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Análise de Sequência com Séries de Oligonucleotídeos , Oryza/crescimento & desenvolvimento , RNA de Plantas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Especificidade da EspécieRESUMO
AIMS/HYPOTHESIS: Insulin activates insulin receptor protein tyrosine kinase and downstream phosphatidylinositol-3-kinase (PI3K)/Akt signalling in muscle to promote glucose uptake. The insulin receptor can serve as a substrate for the protein tyrosine phosphatase (PTP) 1B and T cell protein tyrosine phosphatase (TCPTP), which share a striking 74% sequence identity in their catalytic domains. PTP1B is a validated therapeutic target for the alleviation of insulin resistance in type 2 diabetes. PTP1B dephosphorylates the insulin receptor in liver and muscle to regulate glucose homeostasis, whereas TCPTP regulates insulin receptor signalling and gluconeogenesis in the liver. In this study we assessed for the first time the role of TCPTP in the regulation of insulin receptor signalling in muscle. METHODS: We generated muscle-specific TCPTP-deficient (Mck-Cre;Ptpn2(lox/lox)) mice (Mck, also known as Ckm) and assessed the impact on glucose homeostasis and muscle insulin receptor signalling in chow-fed versus high-fat-fed mice. RESULTS: Blood glucose and insulin levels, insulin and glucose tolerance, and insulin-induced muscle insulin receptor activation and downstream PI3K/Akt signalling remained unaltered in chow-fed Mck-Cre;Ptpn2(lox/lox) versus Ptpn2(lox/lox) mice. In addition, body weight, adiposity, energy expenditure, insulin sensitivity and glucose homeostasis were not altered in high-fat-fed Mck-Cre;Ptpn2(lox/lox) versus Ptpn2(lox/lox) mice. CONCLUSIONS/INTERPRETATION: These results indicate that TCPTP deficiency in muscle has no effect on insulin signalling and glucose homeostasis, and does not prevent high-fat diet-induced insulin resistance. Thus, despite their high degree of sequence identity, PTP1B and TCPTP contribute differentially to insulin receptor regulation in muscle. Our results are consistent with the notion that these two highly related PTPs make distinct contributions to insulin receptor regulation in different tissues.
Assuntos
Glucose/metabolismo , Músculos/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 2/deficiência , Proteína Tirosina Fosfatase não Receptora Tipo 2/fisiologia , Animais , Diabetes Mellitus Tipo 2/sangue , Teste de Tolerância a Glucose , Homeostase , Insulina/metabolismo , Resistência à Insulina , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 2/genética , Receptor de Insulina/metabolismo , Transdução de Sinais , Fatores de Tempo , Distribuição TecidualRESUMO
Typically, production of induced pluripotent stem cells requires direct contact with feeder cells. However, once the stem cells have reached the appropriate maturation point, it is difficult to separate them from feeder cells, which must be irradiated with γ-rays or treated with the antibiotic mitomycin-C. We used a microporous poly-membrane-based indirect contact co-culture system with mouse embryonic fibroblasts to induce mouse pluripotent stem cells without radiation or antibiotics. We found that induced pluripotent stem cells induced by this co-culture method had a reprogramming efficiency and time similar to those induced using traditional methods. Furthermore, strongly expressed pluripotent markers showed a normal karyotype and formation and contained all three germ layers in a teratoma.
Assuntos
Células-Tronco Embrionárias/fisiologia , Células-Tronco Pluripotentes Induzidas/fisiologia , Animais , Antígenos de Diferenciação/metabolismo , Diferenciação Celular , Técnicas de Cocultura/instrumentação , Técnicas de Cocultura/métodos , Células Alimentadoras , Fibroblastos/fisiologia , Proteínas de Homeodomínio/metabolismo , Células-Tronco Pluripotentes Induzidas/transplante , Cariótipo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Proteína Homeobox Nanog , Teratoma/patologia , Fatores de Transcrição/metabolismoRESUMO
Intrathymic injection is a common technique used for research concerning immunotolerance induction, gene therapy and T cell development in mice. Traditionally used protocols involve major surgery that exposes the thoracic cavity, which results in injury to the mice and increased risk of poor recovery and postsurgical complications such as infection. We introduce a simplified intrathymic injection technique that does not expose the thoracic cavity and virtually eliminates pain, distress and postoperative complications while maintaining high injection efficiency. The technique is suitable for both adult and neonatal mice.
Assuntos
Injeções/métodos , Timo/anatomia & histologia , Animais , Camundongos , Camundongos Endogâmicos C57BL , Agulhas , SeringasRESUMO
WHAT IS KNOWN AND OBJECTIVE: The Department of Health (DOH) in Taiwan issued the 'Guidelines for Benzodiazepine Use in Sedation and Hypnosis' in March 2004, which clearly stated that benzodiazepines (BZDs) should not be used alone for the treatment of depression. However, the extent to which clinicians comply with the BZD guidelines was not known. This study aimed to evaluate whether sole prescribing of BZDs for major depression decreased after the implementation of the BZD guidelines. METHODS: This was a retrospective longitudinal trend analysis by analyzing the Longitudinal Health Insurance Database (LHID) from September 2002 to September 2005. The LHID contains all claims data from a random sample of 1,000,000 beneficiaries of the universal National Health Insurance programme in Taiwan. The 3-year study period was divided equally into six periods, before and after the implementation of the guidelines respectively. For each period, the proportion of patients with major depression (ICD-9-CM code 296.2x, 296.3x) treated with BZDs without any concomitant antidepressant was calculated in order to conduct a trend analysis. RESULTS AND DISCUSSION: A total of 5463 prescriptions of BZDs solely used for major depression were observed in the entire study period. In more than 80% of the BZD prescriptions in which BZDs were used alone for major depression, they were prescribed at doses higher than one prescribed daily dose/defined daily dose and were supplied for more than 7 days. The number of outpatients with major depression ranged from 2137 to 3326 during the 12 periods. The proportion of depressed patients treated with BZDs alone per 3 months (i.e., the non-compliance rate) fluctuated from 6·7% to 9·4% before implementation of the guidelines, and from 8·0% to 9·4% after implementation, in outpatient settings. In addition, the guideline non-adherence rates in inpatient settings varied from 7·0% to 11·8% and from 7·8% to 12·6% before and after the implementation of the BZD guidelines respectively. Further trend analyses indicated that the implementation of the guidelines was not associated with a reduced rate of sole prescribing of BZDs for major depression in either inpatient (P = 0·083) or outpatient settings (P = 0·925). WHAT IS NEW AND CONCLUSION: The formulation and implementation of the BZD guidelines appear not to be associated with a reduced rate of sole prescribing of BZDs for major depression, and more comprehensive efforts are required.