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Long non-coding RNAs (lncRNAs) play critical roles in the various physiological processes of insects. The wing is a successful adaptation allowing insects to escape from unfavorable environments, while information on lncRNAs related to wing development is limited. In this study, we constructed 12 libraries from two RNA-seq comparisons: 4th instar winged nymphs versus winged adults and 4th instar wingless nymphs versus wingless adults in the brown citrus aphid Aphis citricidus, to identify the wing development-associated lncRNAs. A total of 2914 lncRNAs were identified and 50 lncRNAs were differentially expressed during the 4th instar winged nymphs to winged adults transition, and 28 lncRNAs changed during the 4th instar wingless nymphs to wingless adults transition. The differentially expressed lncRNAs were grouped into six clusters according to the expression patterns in the combined two-winged morphs. lncRNA Ac_lnc54106.1 was up-regulated during 4th instar winged nymphs to winged adults transition, but a lack of change during the 4th instar wingless nymphs to wingless adults transition implied a critical role in the specific regulation of wing development. RNA interference of Ac_lnc54106.1 resulted in malformed wings. Targets prediction, expression patterns, and RNAi assay results showed that Ac_lnc54106.1 may target the PiggyBac transposable element-derived protein 4 (PGBD4) gene, decrease expression of the canonical wing development-related genes, and finally regulate wing development. The systematic identification of lncRNAs in an aphid increases our understanding of how non-coding RNA mediates the wing plasticity of insects.
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Afídeos/crescimento & desenvolvimento , Afídeos/genética , Regulação da Expressão Gênica no Desenvolvimento , RNA Longo não Codificante/genética , Asas de Animais/crescimento & desenvolvimento , Animais , Perfilação da Expressão Gênica , Ninfa/genética , Ninfa/crescimento & desenvolvimento , RNA Longo não Codificante/metabolismoRESUMO
Cutaneous blastomycosis is endemic to North America and is often caused by dimorphic fungi with spores that are inhaled, inoculated spores, or hyphae that infect immunosuppressed and healthy people. It is sporadic and described as a universal imitator with morphological manifestations as erythema, nodules, and ulcers. Our case demonstrated a 69-year-old female bitten by her pet dog who was then diagnosed with cutaneous blastomycosis through social history and detailed laboratory examinations. She experienced a prolonged failure with antibacterial treatment, negative stool and tissue culture, and chronic inflammatory cell infiltrates on tissue pathology. High-throughput sequencing was performed and showed evidence of Blastomyces dermatitidis aetiology. Photodynamic therapy combined with oral itraconazole was administered, and the patient recovered in a short time. Our case presents inoculated cutaneous blastomycosis and a treatment approach in which photodynamic therapy combined with oral itraconazole significantly reduced the duration of disease treatment and affords a promising choice for the treatment of cutaneous blastomycosis.
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Blastomicose , Fotoquimioterapia , Animais , Antifúngicos/uso terapêutico , Blastomyces , Blastomicose/tratamento farmacológico , Cães , Feminino , Humanos , Itraconazol/uso terapêutico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/uso terapêuticoRESUMO
A cascade alkynylation and selective hydrogenation catalyzed by covalent binaphthyl-stabilized palladium nanoparticles has been established, providing a novel and highly efficient methodology for accessing Z and Z,Z-selective phosphinyl [3]dendralenes with broad functional group tolerance and good yields. This strategy achieves the first cascade reactions of alkynylation and hydrogenation with chemoselectivity modulated by catalyst loading.
RESUMO
A cost-effective system is revealed here to construct polyfunctionalized 2H-pyran cores containing phosphinyl groups with an organobase as a catalyst. Good to excellent yields were obtained under the optimized reaction conditions, with a broad substrate scope tolerated.
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BACKGROUND: Atrial tachyarrhythmia (AT) is a common complication in patients who have undergone a Fontan operation. In this study, we investigated whether abnormal Ca2+ handling contributes to the Fontan operation-related atrial arrhythmogenic substrate. METHODS: Mongrel dogs were randomly assigned to sham and Fontan groups. The Fontan operation model was developed by performing an atriopulmonary anastomosis. After 14 days, an electrophysiological study was performed to evaluate the AT vulnerability. Ca2+ handling properties were measured by loading atrial cardiomyocytes (CMs) with fura-2 AM. The L-type Ca2+ (ICa-L) and Na+-Ca2+ exchanger (INCX) currents of the CMs were recorded by the whole-cell patch-clamp technique. The key Ca2+ handling proteins expression was assessed by Western blotting. RESULTS: The AT inducibility was higher in the Fontan group than in the sham group (85.71 vs. 14.29%, P < 0.05). The Fontan operation resulted in decreased Ca2+ transient (CaT) amplitude and sarcoplasmic reticulum (SR) Ca2+ content, but in enhanced diastolic intracellular Ca2+ concentration and SR Ca2+ leak in the atrial CMs. The spontaneous CaT events, triggered ectopic activity and INCX density were increased, but ICa-L density was reduced in CMs from the Fontan atria (all P < 0.05). Additionally, the Fontan operation resulted in decreased SR Ca2+ ATPase expression and Cav1.2 expression, but in increased NCX1 and Ser2814-phosphorylated ryanodine receptor 2. The calmodulin-dependent protein kinase II expression and function were markedly enhanced in the Fontan atria. CONCLUSION: The Fontan operation caused atrial CM Ca2+ handling abnormalities that produced arrhythmogenic-triggered activity and increased vulnerability to AT in experimental Fontan dogs.
Assuntos
Fibrilação Atrial/etiologia , Cálcio/metabolismo , Técnica de Fontan/efeitos adversos , Miocárdio/metabolismo , Animais , Cães , Fura-2/análogos & derivados , Fura-2/farmacologia , Átrios do Coração , Modelos Animais , Miócitos Cardíacos/metabolismo , Técnicas de Patch-Clamp , Retículo Sarcoplasmático/metabolismo , Trocador de Sódio e Cálcio/metabolismoRESUMO
PURPOSE: Skin ulcer is a common type of disease affecting patients' health and quality of life, and bacterial infection increases the difficulty of its management. METHODS: The present study collected the results of bacterial culture sampled from the surface of 110 cases of skin ulcers at our hospital from January 2011 to December 2012. We analyzed the constituent ratios of ulcer surface bacteria, the change in the main infectious bacteria and the results of drug-sensitivity testing for common bacteria. In addition, the characteristics of bacterial infection of skin ulcers were summarized. RESULT: Of the 110 samples, 90 isolated bacteria were cultured. Sixty-one were Gram-negative bacteria, mainly comprising Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli. In addition, 23 isolates were Gram-positive bacteria, mainly comprising Staphylococcus aureus and Enterococcus faecalis. The probability of a negative bacterial culture in 2012 was significantly lower than that in 2011 (16.7% vs. 40.0%, p < 0.01). Moreover, the probability of P. aeruginosa infection in 2012 was significantly higher than that in 2011 (31.7% vs. 14.0%, p < 0.01). P. aeruginosa was resistant to seven commonly used antibiotics. Both K. pneumoniae and E. coli had higher resistance to ampicillin. E. cloacae were not sensitive to piperacillin/tazobactam. Acinetobacter baumannii was resistant to all the tested drugs. S. aureus, E. faecalis and Staphylococcus epidermidis had high resistance to clindamycin. There was other drug resistance to reflect the higher rate of skin bacterial resistance. CONCLUSION: Skin bacterial resistance rate is high. Gram-negative bacteria gradually account for the majority, and P. aeruginosa becomes the most important skin infection pathogen. These characteristics of bacterial infections of skin ulcers provide a significant reference for guiding the selection of antibiotics, better controlling infections of skin ulcers and accelerating the healing of skin ulcers.
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Antibacterianos/uso terapêutico , Bactérias/efeitos dos fármacos , Infecções Bacterianas/tratamento farmacológico , Úlcera Cutânea/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Úlcera Cutânea/microbiologia , Adulto JovemRESUMO
BACKGROUND: Helicobacter pylori (H. pylori) seem to involve in the etiology of chronic spontaneous urticaria (CSU). But studies of the pathogenic mechanism are very little. METHODS: In this study, we detected the serum-specific anti-H. pylori IgG and IgE antibodies in 211 CSU and 137 normal subjects by enzyme-linked immunosorbent assay (ELISA), evaluated the direct activation effects of H. pylori preparations and its protein components on human LAD2 mast cell line in vitro, and analyzed the specific protein ingredients and functions of the most effective H. pylori mixed protein component using liquid chromatography-mass spectrometry and ELISA assay. RESULTS: In CSU patients, the positive rate of anti-H. pylori IgG positive rate was significantly higher than that in normal controls, and the anti-H. pylori IgE levels had no statistical difference between H. pylori-infected patients with and without CSU. Further studies suggested that H. pylori preparations can directly activate human LAD2 mast cell line in a dose-dependent manner and its most powerful protein component was a mixture of 21-35 kDa proteins. Moreover, the 21-35 kDa mixed protein component mainly contained 23 kinds of proteins, which can stimulate the release of histamine, TNF-a, IL-3, IFN-γ, and LTB4 by LAD2 cells in a dose-dependent or time-dependent manner. CONCLUSIONS: A 21-35 kDa mixed protein component should be regarded as the most promising pathogenic factor contributing to the CSU associated with H. pylori infection.
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Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Helicobacter pylori/imunologia , Mastócitos/imunologia , Urticária/etiologia , Adolescente , Adulto , Idoso , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/química , Proteínas de Bactérias/química , Linhagem Celular , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Peso Molecular , Análise de Sequência de DNA , Soro/imunologia , Urticária/patologia , Adulto JovemRESUMO
OBJECTIVE: The purpose of this study was to evaluate the treatment outcome and safety of topical photodynamic therapy (PDT) with aminolevulinic acid (5-ALA) combined with surgical curettage for perianal condyloma acuminata. BACKGROUND DATA: Condyloma acuminata is the most common sexually transmitted disease, with a high relapse rate, especially in the perianal area. The outcomes of many treatment methods for perianal genital warts are not satisfactory. METHODS: In this study, 40 cases of perianal condyloma acuminata were treated with topical aminolevulinic acid photodynamic therapy (ALA-PDT) combined with curettage. First, the warts were removed with the patient under local anesthesia. The depth of the ablation and curettage reached the dermal layer. Then, the first session of PDT was performed after 2 days. Ten percent 5-ALA cream was applied to lesional skin with occlusive dressing for a 3-h incubation period and the lesions of warts were irradiated for 10 min by using a 635-nm laser beam of 177 mW/cm(2) intensity. The ALA-PDT was repeated after 1 and 3 weeks. Follow-up evaluation was performed at our outpatient clinic at 1 and 3 months after completion of therapy. Before treatment and at each follow-up visit, the lesions were photographed; the treatment outcome was evaluated as recurrence rate, and patient satisfaction was surveyed. RESULTS: After three PDT sessions following surgical curettage, all 40 patients were cured and there was no recurrence at 1 month off treatment. At 3 months off treatment, six cases relapsed, corresponding to a recurrent rate of 15%. The satisfaction rate of patients was 100% at 1 month and 95% at 3 months after treatment. CONCLUSIONS: Combination of surgical curettage and topical PDT has been proven to be a safe and effective procedure, and may offer a wide clinical application for the treatment of perianal condyloma acuminata.
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Ácido Aminolevulínico/uso terapêutico , Canal Anal/patologia , Condiloma Acuminado/tratamento farmacológico , Condiloma Acuminado/cirurgia , Curetagem/métodos , Fármacos Fotossensibilizantes/uso terapêutico , Administração Tópica , Adulto , Estudos de Coortes , Terapia Combinada , Condiloma Acuminado/diagnóstico , Feminino , Seguimentos , Humanos , Masculino , Estudos Prospectivos , Medição de Risco , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
OBJECTIVES: To evaluate the safety and effect of systemic administration of photosensitizer Photocarcinorin (PsD-007) in the treatment of oral florid papillomatosis (OFP). BACKGROUND: OFP is a rare and refractory disease characterized by multiple papillomatous lesions. The outcomes of many methods for OFP are not satisfactory. MATERIALS AND METHODS: Photosensitizer Photocarcinorin (5 mg/kg) was intravenously injected into a 59-year-old woman with widespread OFP, and the lesions were irradiated for 10 min by using a 635-nm laser beam of 177 mW/cm(2) intensity and 106.2 J/cm(2) energy density. The therapy was performed once a month for 3 months in total. RESULTS: After three courses, the small lesions disappeared, and the larger ones were reduced by ≥80%. The adverse effects were minimal. CONCLUSIONS: Systemic PDT is an effective and successful treatment modality for OFP, especially in elderly patients.
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Hematoporfirinas/uso terapêutico , Neoplasias Bucais/tratamento farmacológico , Papiloma/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: To evaluate the safety and efficacy of topical aminolevulinic acid (ALA) photodynamic therapy (PDT) in the treatment of verruca planae. METHODS: A total of 18 patients with facial verruca planae was treated with ALA-PDT. A 10% ALA emulsion was applied on facial lesions for a 4-h incubation period after azone infiltrating for 30 min. Skin lesions of verruca planae were irradiated with 120 J/cm(2) using a 635-nm laser for 10 min. Each patient received at least two sessions at intervals of 15 days. Those with residual lesions after the first session were treated with a third PDT. Before treatment and at each return visit, the lesions were photographed. Patient satisfaction was investigated and therapeutic effect was evaluated at 6 months after termination of the therapy. RESULTS: Fifteen days after the first topical ALA-PDT, 10 patients achieved complete response and 7 patients showed improvement. Complete clinical response was observed in 17 patients after two sessions. At 6 months after termination, only one case relapsed among the complete clinical response patients, and the patient satisfaction rate was 88.89%. CONCLUSION: PDT, utilizing the topical application of 10% ALA, has a good result and excellent cosmetic effects for the treatment of verruca planae.
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Ácido Aminolevulínico/administração & dosagem , Dermatoses Faciais/terapia , Fotoquimioterapia , Fármacos Fotossensibilizantes/administração & dosagem , Verrugas/terapia , Administração Tópica , Adolescente , Adulto , Feminino , Humanos , MasculinoRESUMO
OBJECTIVES: The purpose of this study was to observe the improvements of port-wine stains (PWSs) under photodynamic therapy (PDT) and to further evaluate the safety and efficacy of PDT in the dermatology clinic for the treatment of PWSs. METHODS: Total of 75 PWS patients were treated with PDT. The PWS lesions were exposed to the copper vapor laser after intravenous injection of photosensitizer (Photocarcinorin, PsD-007). The lesions were photographed before each session and the therapeutic effects were evaluated by physicians based on the improvements in color, thickness and area of the photographed lesions. A follow-up visit was carried out after 2 months. The improvement rates were classified into 5 grades for quantitation of the therapeutic effects. A decision of termination or continuation of the treatment was then made according to lesion changes. RESULTS: Patients with PWSs responded remarkably to PDT. The complete clinical remission rate was 57.33% and the effective rate was up to 94.67% in no more than four courses of treatment. Among the completely responded patients, 20.93% were treated with no more than two courses. CONCLUSION: PDT is safe and highly effective for the treatment of PWSs in the dermatology clinic.
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Hematoporfirinas/farmacologia , Fotoquimioterapia/métodos , Mancha Vinho do Porto/tratamento farmacológico , Adolescente , Adulto , Criança , Pré-Escolar , Estudos de Coortes , Estética , Dermatoses Faciais/diagnóstico , Dermatoses Faciais/tratamento farmacológico , Feminino , Seguimentos , Humanos , Injeções Intravenosas , Masculino , Satisfação do Paciente , Fotografação , Mancha Vinho do Porto/diagnóstico , Estudos Retrospectivos , Índice de Gravidade de Doença , Resultado do Tratamento , Adulto JovemRESUMO
By comparing the drug distribution of breviscapine administered intranasally, orally and intrgvenous injected in rats' brain. After 0.4 mg x kg(-1) breviscapine was given by tail vein, intranasal and gastric perfusion administration to SD rats, cerebrospinal fluid was obtained by erebellomedllery cisternal puncture at different times. 125I labeling was used to determine the drug content of cerebrospinal fluid, cerebrum, cerebellum, medulla oblongata, olfactory region, olfactory bulb and blood in rats. AUCs were calculated by trapezoidal rule. The results showed that AUCs(0-240 min) (microg x min x g(-1)) of brain tissues were 11.686 +/- 1.919, 5.676 +/- 1.025, 7.989 +/- 0.925, 7.956 +/- 1.159, 17.465 +/- 2.136, 24.2 +/- 2.906 and 78.51 +/- 12.05, respectively, in the intranasal administration group; while those in the tail vein administration groups were 6.79 +/- 0.661, 6.251 +/- 0.40, 10.805 +/- 1.161, 9.146 +/- 1.04, 9.892 +/- 1.532, 7.871 +/- 0.842 and 173.91 +/- 10.02; and oral administration group were 0.868 +/- 0.167, 1.708 +/- 0.266, 2.867 +/- 0.725, 2.067 +/- 0.313, 1.361 +/- 0.308, 1.206 +/- 0.255 and 45.2 +/- 7.52, respectively. AUCs(0-240 min) of the brain tissues after oral, tail vein and intranasal administration were 22.29%, 29.18%, 95.49% of that of blood, respectively, it means that the absorption rate and drug distribution in the brain tissues after intranasal administration were higher than those of oral and tail vein administration. It is worth to investigate further the pharmacodynamic relationship.
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Encéfalo/metabolismo , Flavonoides/farmacocinética , Administração Intranasal , Administração Oral , Animais , Área Sob a Curva , Cerebelo/metabolismo , Cérebro/metabolismo , Sistemas de Liberação de Medicamentos , Erigeron/química , Flavonoides/administração & dosagem , Flavonoides/sangue , Flavonoides/líquido cefalorraquidiano , Injeções Intravenosas , Masculino , Bulbo/metabolismo , Bulbo Olfatório/metabolismo , Condutos Olfatórios/metabolismo , Plantas Medicinais/química , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
OBJECTIVE: To observe the influence of Wnt-1 recombinant adenovirus on differentiation tendency of human epidermal stem cells. METHODS: Wnt-1 recombinant adenovirus was transduced into hESCs (E group), while normal hESCs were used as control (C) group. The diameter, proliferation,and labeling molecular expression of hESC were determined. The content of MMP-2 and MMP-7 in supernate were also assayed. RESULTS: There was no obvious difference in diameter of hESC between two groups. The density of hESC in E group was (1.45 +/- 0.09) x 10(5)/mL, which was obviously higher than that in C group [(1.18 +/- 0.10) x 10(5)/mL, P < 0.05]. There were no obvious differences in expression of markers between two groups,including keratin 5 (KS), K6, K7, KS, K14, CD44, carcinoembryonic-like antigen (CEAA), ER, PR (P > 0.05) ,while the expression of K 10 was different among groups [(60 +/- 3)% in E group, 0 in C group], also K18 [(34.3 +/- 2.1)% in E group vs. (13.8 +/- 1.7)% in C group, P < 0.05], and K19 [(17.1 +/- 1.8)% in E group vs. (24.4 +/- 1.5)% in C group, P < 0.05].The contents of MMP-2 and MMP-7 in E group were higher than those in C group (P < 0.01). CONCLUSION: Wnt-1 recombinant adenovirus can induce the differentiation of hESCs to glandular epithelium-like cells.
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Adenoviridae/genética , Diferenciação Celular , Células Epiteliais/citologia , Células-Tronco/citologia , Proteína Wnt1/genética , Linhagem Celular , Células Epiteliais/virologia , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 7 da Matriz/metabolismoRESUMO
OBJECTIVE: This study was designed to compare clinical efficacy of segmental pulmonary vein ablation (SPVI), amiodarone or amiodarone plus losartan on sinus rhythm maintenance in patients with lone paroxysmal atrial fibrillation (PAF). METHODS: Patients with lone PAF were treated with amiodarone alone (A, n = 52), segmental pulmonary vein isolation (SPVI, n = 51), or amiodarone plus losartan (AL, n = 51). The primary endpoint of this study was the incidence of symptomatic atrial tachyarrhythmia (> 30 s) documented by 12 lead ECG or Holter during 12 months follow-up period. RESULTS: During follow-up, AF was documented in 24 patients (46.2%) in A group, 11 patients (21.6%) in SPVI group and 12 (23.5%) in AL group (P < 0.05 vs. A group). The Kaplan-Meier survival analysis demonstrated a significant equally reduction in AF recurrence in SPVI and AL groups (P = 0.009, log-rank test and P = 0.018, log-rank test, respectively) compared with A group. The hazard ratio for AF recurrence in patients treated with SPVI and amiodarone plus losartan was 0.41 (95% CI 0.200 to 0.848, P = 0.016) and 0.46 (95% CI 0.225 to 0.953, P = 0.036), respectively. Incidences of major adverse cardiac events were similar among the groups (9.6% in A, 3.9% in SPVI and 7.8% in AL group, P > 0.05). CONCLUSION: The results of this study suggest that the segmental pulmonary vein isolation and amiodarone plus losartan are superior to amiodarone alone for preventing AF recurrence in patients with lone PAF.
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Amiodarona/uso terapêutico , Fibrilação Atrial/terapia , Losartan/uso terapêutico , Idoso , Antiarrítmicos/uso terapêutico , Fibrilação Atrial/prevenção & controle , Ablação por Cateter/métodos , Terapia Combinada , Seguimentos , Humanos , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do TratamentoRESUMO
AIM: To clone HPV58 E6 gene and analyze the HLA-DQB1 03-restricted T cell epitopes on E6 protein. METHODS: Total genome DNA was isolated from a cervical cancer sample. The HPV58 E6 gene was amplified by PCR and inserted into pGEM-T Easy vector. The recombinant plasmid was identified by restriction endonuclease analysis and sequencing. HLA-DQB1 03-restricted T cell epitopes on E6 protein were predicted and analyzed by the position-specific scoring matrix, support vector machine theory and prediction algorithm for proteasomal cleavages. RESULTS: A HPV58 E6 gene was successfully cloned and submitted to GenBank (EF060239). Epitope 47(FADLRIAYRDGNPFA) and Epitope 102(RCIICQRPLCPQEKK) were theoretic HLA-DQB1 03-restricted T cell epitopes on E6 protein. CONCLUSION: These two epitopes could serve as candidates for screening and identification of the vaccine against HPV58 infection in the further study.
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Alphapapillomavirus/genética , Epitopos de Linfócito T/análise , Epitopos de Linfócito T/imunologia , Antígenos HLA-DQ/análise , Antígenos HLA-DQ/imunologia , Proteínas Virais/genética , Adulto , Alphapapillomavirus/imunologia , Sequência de Aminoácidos , Sequência de Bases , Linhagem Celular Tumoral , Clonagem Molecular , Epitopos de Linfócito T/química , Feminino , Antígenos HLA-DQ/química , Humanos , Dados de Sequência Molecular , Vacinas contra Papillomavirus/análise , Vacinas contra Papillomavirus/imunologia , Plasmídeos/genética , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Complexo de Endopeptidases do Proteassoma/metabolismo , Alinhamento de Sequência , Proteínas Virais/imunologiaRESUMO
OBJECTIVE: To observe the histological characteristics of constructed basement membrane in tissue-engineered skin. METHODS: Forskins from circumcision in normal children were obtained with informed consent of the parents, and then the epidermal keratinocytes (KC) and dermal fibroblasts (Fb) were isolated with trypsin and collagenase D digestion in sequence. Tissue engineered skin with composite chitosan was maintained in a submerged state for 3 days, and then at the air-liquid interface. The tissue-engineered skins were fixed in neutral formalin and then embedded in paraffin after culture for 7, 10 and 15 days, respectively for immunohistological examination of the basement membrane component,including the condition of collagen type IV (COL-IV), collagen type VII (COL-VII), and laminin (LN). RESULTS: HE staining showed that the keratinocytes formed a fine stratified squamous epithelium with the presence of basal, spinous, granular and corneous cell layers, and there was various amount of cells in flat and fusiform shape in each layer. It was found that a regular red staining strip situated at the dermal epidermal junction. Positive staining of collagen IV, collagen VII as well as LN was observed by immunohistological examination. CONCLUSION: The results suggest that the composite chitosan tissue engineered skin has a good prospect for clinical use because it presents a perfect reconstruction of basement membrane.
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Membrana Basal/citologia , Pele Artificial , Engenharia Tecidual/métodos , Células Cultivadas , Criança , Quitosana/metabolismo , Colágeno Tipo IV/metabolismo , Colágeno Tipo VII/metabolismo , Humanos , Laminina/metabolismo , Técnicas de Cultura de ÓrgãosRESUMO
To investigate the possibility of hair follicle reformation induced by dermal papilla cells in vivo and in vitro. Dermal papilla cells, dermal sheath cells obtained from human scalp skin by enzyme digestion were mixed with collagen to form mesenchymal cell-populated collagen gels. Superior and inferior epithelial cells and bulb matrical cells were then cultured on these gels by organotypic culture to recombine bilayer artificial skins. Dermal papilla cells and outer root sheath keratinocytes were mingled together and transplanted under subcutaneous tissue of the dorsal skin of nude mice. The results of histologic examination was observed with HE stain. These recombinants by organotypic culture all reformed bilayer structure like nature skin. Hair follicle-like structure reformation was found in dermal sheath cell-populated collagen gel when combined with superior or inferior epithelial cells. Dermal papilla cells also induced superior and inferior epithelial cells to form hair follicle on nude mice. Low passage dermal papilla cells mixed with hair follicle epithelial cells reformed many typical hair follicle structures and produced hair fibres after transplantation on nude mice. The dermal part of hair follicle, such as dermal papilla cells and dermal sheath cells, has the ability to induce hair follicle formation by interaction with the epithelial cells of hair follicle.
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Transplante de Células/fisiologia , Derme/citologia , Folículo Piloso/crescimento & desenvolvimento , Couro Cabeludo/citologia , Animais , Comunicação Celular , Técnicas de Cultura de Células , Transplante de Células/métodos , Células Cultivadas , Folículo Piloso/anatomia & histologia , Histocitoquímica , Humanos , Camundongos , Camundongos Nus , Couro Cabeludo/fisiologia , Pele Artificial , Engenharia Tecidual/métodosRESUMO
OBJECTIVE: The purpose of the present study was to evaluate the clinical efficacy of perindopril or losartan in combination with low-dose amiodarone on maintenance of sinus rhythm in patients with idiopathic paroxysmal atrial fibrillation (PAF). METHODS: One hundred and eighty-one patients with idiopathic PAF were included in the study and randomly divided into three groups: group 1 (amiodarone group, n = 61) was treated with amiodarone alone, group 2 (amiodarone plus losartan, n = 59) was treated with amiodarone and perindopril in combination, and group 3 (amiodarone plus perindopril group, n = 61) was treated with amiodarone and perindopril in combination. The left atrial diameter (LAD) was measured with transthoracic echocardiogram at before and after 6, 12, 18 and 24-month of treatment. The duration of observation was up to two years and the primary end point of the study was the first recurrence of AF. RESULTS: During the 6 month following up, there was no difference in LAD among the three groups. After 12 months, LAD in group 1 was significantly larger than group 2 and group 3 (P < 0.05). At 7th-month, the sinus rhythm maintenance of group 1 was lower significantly than group 2 and group 3. At the end of the study, the maintenance of sinus rhythm in group 2 and group 3 was higher significantly than in group 1 (83.05% and 80.33% vs 59.01%, P < 0.05), nevertheless, there was no significant difference between group 2 and group 3. CONCLUSIONS: The results of this study suggest that the combination of amiodarone with angiotensin converting enzyme inhibitor perindopril or with angiotensin II receptor antagonist losartan are more effective than amiodarone alone in sinus rhythm maintenance for idiopathic PAF. ACEI and ARB can inhibit the enlargement of left atrium and reduce recurrence rate in patients with idiopathic PAF.
Assuntos
Amiodarona/uso terapêutico , Antiarrítmicos/uso terapêutico , Fibrilação Atrial/tratamento farmacológico , Losartan/uso terapêutico , Perindopril/uso terapêutico , Adulto , Idoso , Quimioterapia Combinada , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
BACKGROUND: Dermal papilla cells (DPC) are a group of mesenchyme-derived cells at the base of the hair follicle, where they regulate and control hair follicle growth through the expression and secretion of cytokines. Nevertheless, the role of DPC derived chemokines and other cytokines in the hair follicle biology remain speculative. In this study, we investigated the expression of basic fibroblast growth factor (bFGF), endothelin-1 (ET-1) and stem cell factor (SCF) in different passages of cultured DPC and their effects on the biological behaviour of DPC. METHODS: The expression of bFGF, ET-1 and SCF in different passages of cultured DPC and their possible effects on the biological behavior of DPC are investigated using in situ hybridization and immunochemistry. In addition, we performed transplantation of hair follicle cells into nude mice. The cultured DPC, dermal sheath cells and fibroblast of human scalp, respectively, were mixed with cells of the hair follicle epithelium in different ratios, and then were cultured in hair follicle organotypic cultures or implanted into the subcutis of nude mice. RESULTS: The expression of ET-1 and SCF in early passages of cultured DPC became stronger, but turned weaker and even negative in late passages (> 6 passages). Hair follicle-like structures were formed after DPC combined with the cells of hair follicle epithelium cells in hair follicle organotypic cultures. When hair follicle organotypic cultures were implanted into the subcutis of nude mice, the relative intact hair follicles were formed. After the transplantation of hair follicle cells into the nude mice, the hair follicle-like structure was formed in the group that contained DPC mixed with hair follicle epithelium cells. However, no hair follicles were formed in the other two groups. It was found that the higher the expression of ET-1 and SCF in DPC, the stronger the ability of DPC to induce hair follicle regeneration. CONCLUSIONS: The cultured DPC can induce hair follicle regeneration and sustain hair growth in vivo and in vitro. Moreover, the expression of ET-1 and SCF is correlated with the ability of DPC inducing hair follicle regeneration.
Assuntos
Folículo Piloso/fisiologia , Regeneração , Pele/citologia , Animais , Células Cultivadas , Endotelina-1/análise , Fator 2 de Crescimento de Fibroblastos/análise , Humanos , Imuno-Histoquímica , Hibridização In Situ , Camundongos , Camundongos Nus , Pele/química , Fator de Células-Tronco/análiseRESUMO
OBJECTIVE: To investigate the expression of bFGF, ET-1 and SCF in different passages of cultured dermal papilla cells (DPC), and their possible effect on biological behaviour of DPC. METHODS: The expression of bFGF, ET-1 and SCF in different passages of cultured DPC was detected by immunocytochemistry and in situ hybridization. RESULT: The expression of ET-1 and SCF in early passages of cultured DPC was stronger, but became negative in late passages (>6 passages). The stronger the expression of ET-1 and SCF in DPC, the higher ability of DPC to induce hair follicle regeneration. CONCLUSION: The expression strength of ET-1 and SCF is related to the ability of DPC inducing hair follicle regeneration.