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1.
J Cardiothorac Surg ; 17(1): 119, 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35578261

RESUMO

OBJECTIVE: Survival from out-of-hospital cardiac arrest (OHCA) often depends on the effective and immediate use of automated external defibrillators (AEDs). Given that there have been few studies about AED use in China, the purpose of this study is to investigate the knowledge and attitudes regarding AED use among the Chinese public, then provide an effective suggestion for AED education strategies and legislation. METHOD: The online survey was conducted among Chinese participants of the First Aid eLearning courses in June 2020. RESULT: A total of 2565 (95.00%) surveys were completed, only 23.46% of respondents with non-medical related respondents reported having attended previous AED training courses. Regarding the basic knowledge of AEDs, few respondents (12.28%, n = 315) could answer all four questions correctly. 95.67% (n = 2454) were willing to learn AED use. Even if without the precondition of being skilled in AEDs, the female was more likely to rescue OHCA patients than the male (p = 0.003). Almost all respondents (96.65%) showed a strong willingness to rescue OHCA patients with training in using AEDs. The top four barriers to rescuing OHCA patients were lack of practical performing ability (60.47%), fear of hurting patients (59.30%), inadequate knowledge of resuscitation techniques (44.19%), and worry about taking legal responsibility (26.74%). CONCLUSION: Our study reflects a deficiency of AED knowledge among the general public in China. However, positive attitudes towards rescuing OHCA patients and learning AED use were observed, which indicates that measures need to be taken to disseminate knowledge and use of AEDs.

2.
Bioact Mater ; 16: 173-186, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35386316

RESUMO

Multifunctional scaffolds with host defense peptides designed for regenerative endodontics are desirable nanobiotechnological tools for dentistry. Here, different scaffolds were tested for use during the pulp revascularization process, including poly(vinyl alcohol)-PVA hydrogels or resins, collagen hydrogels and poly(vinyl alcohol) PVA/Chitosan (PVA/CS) nanofibers. Based on time to degradation (21 days), nanofibers were chosen to be incorporated with ciprofloxacin and IDR-1002 (each at 50 mg/g). Nanofibers containing ciprofloxacin and IDR-1002 had anti-biofilm activity against Enterococcus faecalis, Staphylococcus aureus and a multispecies oral biofilm, besides anti-inflammatory activities. The in vivo subcutaneous tissue response to tooth fragments filled with nanofibers demonstrated a pulp-like tissue formation, when compared to empty teeth fragments. Thus, we designed a strong antimicrobial, immunomodulatory and regenerative candidate for pulp revascularization and regeneration procedures.

3.
BMC Vet Res ; 18(1): 90, 2022 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-35255906

RESUMO

BACKGROUND: Infectious bronchitis virus (IBV) leads to huge economic losses in the poultry industry worldwide. The high levels of mutations of IBV render vaccines partially protective. Therefore, it is urgent to explore an effective antiviral drug or agent. The present study aimed to investigate the in vivo anti-IBV activity of a mixture of plant essential oils (PEO) of cinnamaldehyde (CA) and glycerol monolaurate (GML), designated as Jin-Jing-Zi. RESULTS: The antiviral effects were evaluated by clinical signs, viral loads, immune organ indices, antibody levels, and cytokine levels. The infection rates in the PEO-M (middle dose) and PEO-H (high dose) groups were significantly lower than those in the prevention, positive drug, and PEO-L (low dose) groups. The cure rates in the PEO-M and PEO-H groups were significantly higher than those in the prevention, positive drug, and PEO-L groups, and the PEO-M group had the highest cure rate of 92.31%. The symptom scores and IBV mRNA expression levels were significantly reduced in the PEO-M group. PEO significantly improved the immune organ indices and IBV-specific antibody titers of infected chickens. The anti-inflammatory factor levels of IL-4 and IFN-γ in the PEO-M group maintained high concentrations for a long time. The IL-6 levels in the PEO-M group were lower than those in prevention, positive drug, and PEO-L groups. CONCLUSION: The PEO had remarkable inhibition against IBV and the PEO acts by inhibiting virus multiplication and promoting immune function, suggesting that the PEO has great potential as a novel anti-IBV agent for inhibiting IBV infection.


Assuntos
Infecções por Coronavirus , Vírus da Bronquite Infecciosa , Óleos Voláteis , Doenças das Aves Domésticas , Vacinas Virais , Animais , Antivirais/farmacologia , Antivirais/uso terapêutico , Galinhas , Infecções por Coronavirus/tratamento farmacológico , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/veterinária , Óleos Voláteis/farmacologia , Óleos Voláteis/uso terapêutico , Óleos Vegetais/farmacologia , Óleos Vegetais/uso terapêutico , Doenças das Aves Domésticas/tratamento farmacológico , Doenças das Aves Domésticas/prevenção & controle , Vacinas Virais/uso terapêutico
4.
J Vet Med Sci ; 83(10): 1608-1619, 2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34470981

RESUMO

A novel avian infectious bronchitis virus (IBV) variant, designated as GX-NN160421, was isolated from vaccinated chicken in Guangxi, China, in 2016. Based on analysis of the S1 gene sequence, GX-NN160421 belonged to the New-type 1 (GVI-1) strain. More importantly, three consecutive nucleotides (AAC) deletions were found in the highly conserved structure gene N. The serotype of GX-NN160421 was different from those of the commonly used vaccine strains. The mortality of the GX-NN160421 strain was 3.33%, which contrasted with 50% mortality in the clinical case, but high levels of virus shedding lasted at least 21 days. In conclusion, the first novel IBV variant with three-nucleotide-deletion in the N gene was identified, and this unique variant is low virulent but with a long time of virus shedding, indicating the continuing evolution of IBV and emphasizing the importance of limiting exposure to novel IBV strains as well as extensive monitoring of new IBVs.


Assuntos
Infecções por Coronavirus , Vírus da Bronquite Infecciosa , Doenças das Aves Domésticas , Animais , Galinhas , China/epidemiologia , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/veterinária , Genótipo , Vírus da Bronquite Infecciosa/genética , Nucleocapsídeo , Nucleotídeos , Filogenia , Doenças das Aves Domésticas/epidemiologia
5.
Life Sci ; 266: 118732, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33160996

RESUMO

AIMS: Chronic cerebral hypoperfusion (CCH) elicits inflammatory response, which contributes to the pathology of cognitive impairment. Several studies demonstrate that the alpha-7 nicotinic acetylcholine receptor (α7nAChR) can be a key component to modulate the inflammatory responses. We have reported previously that acupuncture attenuated cognitive deficits induced by CCH. In present study, whether effect of acupuncture was related to α7nAChR mediated anti-inflammatory pathway in CCH rats was further explored. MAIN METHODS: Acupuncture was performed in CCH rats induced by bilateral common carotid arteries occlusion. Neuronal injury, the activation of microglia, the release of inflammatory cytokines, the expression of α7nAChR, and the activation of JAK2/STAT3 signaling pathways were detected. Cognitive function and central inflammation were evaluated after the intraperitoneal injection of an α7nAChR agonist PNU282987, or intracerebroventricular injection of an α7nAChR antagonist α-bungarotoxin (α-BGT). KEY FINDINGS: We found that there were neuronal damage and inflammation, accompanied with the decreased expressions of α7nAChR in the hippocampus under CCH condition. Acupuncture inhibited neuronal damage, activation of microglia, and inflammatory cytokines. The expressions of α7nAChR, together with its downstream JAK2/STAT3 pathways were up regulated by acupuncture. PNU282987 mimicked the anti-inflammatory and neuroprotective effects as well as the cognitive improvements of acupuncture. Meanwhile, the benefit effects of acupuncture above were blocked by α-BGT. SIGNIFICANCE: It was demonstrated that acupuncture promoted cognitive function and afforded neuroprotective effects against inflammation via activation of α7nAChR and its downstream JAK2-STAT3 pathway in CCH rats. It provides a new insight for acupuncture as an anti-inflammatory intervention for cognitive impairment.


Assuntos
Terapia por Acupuntura/métodos , Isquemia Encefálica/complicações , Circulação Cerebrovascular , Disfunção Cognitiva/terapia , Inflamação/prevenção & controle , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Doença Crônica , Disfunção Cognitiva/etiologia , Disfunção Cognitiva/metabolismo , Disfunção Cognitiva/patologia , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Fármacos Neuroprotetores , Ratos , Ratos Wistar , Transdução de Sinais
6.
IUCrJ ; 7(Pt 5): 934-948, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32939285

RESUMO

In Pseudomonas aeruginosa, an important opportunistic pathogen that causes numerous acute and chronic infections, the hybrid two-component system (TCS) regulates the swarming ability and biofilm formation with a multistep phospho-relay, and consists of hybrid-sensor histidine kinase (HK), histidine-containing phospho-transfer protein (Hpt) and response regulator (RR). In this work, two crystal structures of HptB and the receiver domain of HK PA1611 (PA1611REC) of P. aeruginosa have been determined in order to elucidate their interactions for the transfer of the phospho-ryl group. The structure of HptB folds into an elongated four-helix bundle - helices α2, α3, α4 and α5, covered by the short N-terminal helix α1. The imidazole side chain of the conserved active-site histidine residue His57, located near the middle of helix α3, protrudes from the bundle and is exposed to solvent. The structure of PA1611REC possesses a conventional (ß/α)5 topology with five-stranded parallel ß-sheets folded in the central region, surrounded by five α-helices. The divalent Mg2+ ion is located in the negatively charged active-site cleft and interacts with Asp522, Asp565 and Arg567. The HptB-PA1611REC complex is further modeled to analyze the binding surface and interactions between the two proteins. The model shows a shape complementarity between the convex surface of PA1611REC and the kidney-shaped HptB with fewer residues and a different network involved in interactions compared with other TCS complexes, such as SLN1-R1/YPD1 from Saccharomyces cerevisiae and AHK5RD/AHP1 from Arabidopsis thaliana. These structural results provide a better understanding of the TCS in P. aeruginosa and could potentially lead to the discovery of a new treatment for infection.

7.
Oxid Med Cell Longev ; 2020: 8253904, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32850002

RESUMO

BACKGROUND: It is widely accepted that inflammation may contribute to cognitive impairment in patients with vascular dementia (VD). Our prior clinical researches have reported that acupuncture can alleviate cognitive function in VD, but the underlying mechanisms are still unclear. The purpose of this research was to explore whether acupuncture alleviates cognitive impairment by suppressing the microRNA-93- (miR-93-) mediated Toll-like receptor (TLR) signaling pathway, which triggers inflammatory responses in the central nervous system. METHODS: VD was established by permanent bilateral common carotid artery occlusion in male Wistar rats. Three days after operation, the rats began daily treatment with acupuncture for two weeks. The levels of miR-93, Toll-like receptors (TLR2 and TLR4), intracellular signaling molecules (myeloid differentiation factor 88 (MyD88) and nuclear factor-kappa B (NF-κB)), and inflammatory cytokines were subsequently detected. TLR4 colocalized with neurons, microglia, and astrocytes in the hippocampus was evaluated. Neuroinflammation and cognitive function were determined after intracerebroventricular injection of TLR4 antagonist TAK-242 or agonist lipopolysaccharide (LPS) with or without acupuncture. RESULTS: We found that acupuncture notably repressed the expression of inflammatory cytokines in the hippocampus and plasma of VD rats. The expression of TLR4, but not TLR2, was markedly downregulated by acupuncture, accompanied by a decrease in miR-93 and MyD88/NF-κB signaling pathway activation. The overexpression of TLR4 in microglia, but not in astrocytes and neurons, was reversed by acupuncture. Furthermore, intracerebroventricular injection of TAK-242 had similar effects to acupuncture on inflammation and cognitive function, while LPS injection abolished the beneficial effects of acupuncture. CONCLUSIONS: Taken together, these findings provide evidence that acupuncture attenuates cognitive impairment associated with inflammation through inhibition of the miR-93-mediated TLR4/MyD88/NF-κB signaling pathway in experimental VD. Acupuncture serves as a promising alternative therapy and may be an underlying TLR4 inhibitor for the treatment of VD.


Assuntos
Terapia por Acupuntura , Demência Vascular/terapia , Inflamação/terapia , MicroRNAs/metabolismo , Microglia/patologia , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Receptor 4 Toll-Like/metabolismo , Pontos de Acupuntura , Animais , Cognição/efeitos dos fármacos , Demência Vascular/tratamento farmacológico , Demência Vascular/genética , Demência Vascular/fisiopatologia , Hipocampo/patologia , Hipocampo/fisiopatologia , Masculino , Microglia/efeitos dos fármacos , Microglia/metabolismo , Modelos Biológicos , Fármacos Neuroprotetores/farmacologia , Fármacos Neuroprotetores/uso terapêutico , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/antagonistas & inibidores
8.
J Org Chem ; 85(16): 10479-10487, 2020 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-32668153

RESUMO

A highly efficient approach of visible-light-driven radical difluoromethylation of isocyanides to access a wide variety of difluoromethylated phenanthridines and isoquinolines is herein described. Electrophilic S-(difluoromethyl)diarylsulfonium salt proved to be a good difluoromethyl radical precursor under photoredox catalysis. A broad range of isocyanides were tolerated to furnish the corresponding difluoromethylated phenanthridines, isoquinolines, furo[3,2-c]pyridine, and pyrido[3,4-b]indole in moderate to excellent yields under mild conditions. A plausible mechanism was also proposed.

9.
J Org Chem ; 84(24): 15948-15957, 2019 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-31645096

RESUMO

A facile and practical approach for the difluoromethylation of phenols and thiophenols was described. Making use of the recently developed bench-stable S-(difluoromethyl)sulfonium salt as the difluorocarbene precursor, a wide variety of diversely functionalized phenols and thiophenols were readily converted to their corresponding aryl difluoromethyl ethers in good to excellent yields in the presence of lithium hydroxide. Chemoselectivity of various O,S-nucleophiles toward difluorocarbene was systematically studied, suggesting the reactivity order ArS- > RS-, ArO- > ROH > RO-, ArSH, ArOH, RSH.

10.
Toxicon ; 148: 64-73, 2018 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-29654868

RESUMO

Bee venom is a very complex mixture of natural products extracted from honey bee which contains various pharmaceutical properties such as peptides, enzymes, biologically active amines and nonpeptide components. The use of bee venom into the specific points is so called bee venom therapy, which is widely used as a complementary and alternative therapy for 3000 years. A growing number of evidence has demonstrated the anti-inflammation, the anti-apoptosis, the anti-fibrosis and the anti-arthrosclerosis effects of bee venom therapy. With these pharmaceutical characteristics, bee venom therapy has also been used as the therapeutic method in treating rheumatoid arthritis, amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, liver fibrosis, atherosclerosis, pain and others. Although widely used, several cases still reported that bee venom therapy might cause some adverse effects, such as local itching or swelling. In this review, we summarize its potential mechanisms, therapeutic applications, and discuss its existing problems.


Assuntos
Venenos de Abelha/farmacologia , Venenos de Abelha/uso terapêutico , Abelhas , Terapia por Acupuntura , Animais , Venenos de Abelha/efeitos adversos , Humanos , Mordeduras e Picadas de Insetos
11.
Neurochem Int ; 92: 35-42, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26682902

RESUMO

Hippocampal mitochondrial dysfunction due to oxidative stress has been considered to play a major role in the pathogenesis of vascular dementia (VD). Previous studies suggested that acupuncture could improve cerebral hypoperfusion-induced cognitive impairments. However, whether hippocampal mitochondria are associated with this cognitive improvement remains unclear. In this study, an animal model of VD was established via bilateral common carotid arteries occlusion (BCCAO) to investigate the alterations of cognitive ability and hippocampal mitochondrial function. BCCAO rats showed impairments in hippocampal mitochondrial function, overproduction of reactive oxygen species (ROS) and learning and memory deficits. After two-week acupuncture treatment, BCCAO-induced spatial learning and memory impairments as shown in Morris water maze were ameliorated. Hippocampal mitochondrial respiratory complex enzymes (complex I, II, IV) activities and cytochrome c oxidase IV expression significantly increased, which might contribute to the reduction of hippocampal ROS generation. In addition, acupuncture significantly improve mitochondrial bioenergy parameters such as mitochondrial respiratory control rate and membrane potential not PDH A1 expression. Placebo-acupuncture did not produce similar therapeutic effects. These findings suggested that acupuncture reversed BCCAO-induced hippocampal mitochondrial dysfunction, which might contribute to its prevention on cognitive deficits.


Assuntos
Terapia por Acupuntura/métodos , Demência Vascular/metabolismo , Demência Vascular/terapia , Hipocampo/metabolismo , Doenças Mitocondriais/metabolismo , Doenças Mitocondriais/terapia , Animais , Artérias Carótidas/patologia , Transtornos Cognitivos/etiologia , Transtornos Cognitivos/psicologia , Transtornos Cognitivos/terapia , Demência Vascular/psicologia , Hipocampo/enzimologia , Deficiências da Aprendizagem/etiologia , Deficiências da Aprendizagem/psicologia , Deficiências da Aprendizagem/terapia , Masculino , Aprendizagem em Labirinto , Potencial da Membrana Mitocondrial , Transtornos da Memória/etiologia , Transtornos da Memória/psicologia , Transtornos da Memória/terapia , Doenças Mitocondriais/psicologia , Consumo de Oxigênio , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo
12.
Free Radic Biol Med ; 89: 1077-84, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26546103

RESUMO

Emerging evidence suggests acupuncture could exert neuroprotection in the vascular dementia via anti-oxidative effects. However, the involvement of Nrf2, a master regulator of antioxidant defense, in acupuncture-induced neuroprotection in vascular dementia remains undetermined. The goal of our study was to investigate the contribution of Nrf2 in acupuncture and its effects on vascular dementia. Morris water maze and Nissl staining were used to assess the effect of acupuncture on cognitive function and hippocampal neurodegeneration in experimental vascular dementia. The distribution of Nrf2 in neurons in hippocampus, the protein expression of Nrf2 in both cytosol and nucleus, and the protein and mRNA levels of its downstream target genes NQO1 and HO-1 were detected by double immunofluorescent staining, Western blotting and realtime PCR analysis respectively. Cognitive function and microglia activation were measured in both wild-type and Nrf2 gene knockout mice after acupuncture treatment. We found that acupuncture could remarkably reverse the cognitive deficits, neuron cell loss, reactive oxygen species production, and decreased cerebral blood flow. It was notable that acupuncture enhanced nuclear translocation of Nrf2 in neurons and up-regulate the protein and mRNA levels of Nrf2 and its target genes HO-1 and NQO1. Moreover, acupuncture could significantly down-regulated the over-activation of microglia after common carotid artery occlusion surgery. However, the reversed cognitive deficits, neuron cell loss and microglia activation by acupuncture were abolished in Nrf2 gene knockout mice. In conclusion, these findings provide evidence that the neuroprotection of acupuncture in models of vascular dementia was via the Nrf2 activation and Nrf2-dependent microglia activation.


Assuntos
Terapia por Acupuntura , Antioxidantes/farmacologia , Transtornos Cognitivos/prevenção & controle , Demência Vascular/prevenção & controle , Hipocampo/patologia , Fator 2 Relacionado a NF-E2/fisiologia , Neurônios/patologia , Fármacos Neuroprotetores/farmacologia , Animais , Apoptose , Western Blotting , Células Cultivadas , Transtornos Cognitivos/metabolismo , Transtornos Cognitivos/patologia , Demência Vascular/metabolismo , Demência Vascular/patologia , Imunofluorescência , Hipocampo/metabolismo , Técnicas Imunoenzimáticas , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurônios/metabolismo , Estresse Oxidativo/efeitos dos fármacos , RNA Mensageiro/genética , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais
13.
BMC Complement Altern Med ; 15: 133, 2015 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-25928206

RESUMO

BACKGROUND: Decreased cognition is recognized as one of the most severe and consistent behavioral impairments in dementia. Experimental studies have reported that acupuncture may improve cognitive deficits, relieve vascular dementia (VD) symptoms, and increase cerebral perfusion and electrical activity. METHODS: Multi-infarction dementia was modeled in rats with 3% microemboli saline suspension. Two weeks after acupuncture at Zusanli (ST36), all rats were subjected to a hidden platform trial to test their 3-day spatial memory using the Morris water maze test. To estimate the numbers of pyramidal neuron, astrocytes, and synaptic boutons in hippocampal CA1 area, we adopted an unbiased stereology method to accurately sample and measure the size of cells. RESULTS: We found that acupuncture at ST36 significantly decreased the escape latency of VD rats. In addition, acupuncture significantly increased the pyramidal neuron number in hippocampal CA1 area (P < 0.05) and tended to decrease the number of astrocytes (P = 0.063). However, there was no significant change in the synaptic bouton number of hippocampal CA1 area in any of the groups (P > 0.05). CONCLUSIONS: These findings suggest that acupuncture may improve cognitive deficits and increase pyramidal neuron number of hippocampal CA1 area in VD rats.


Assuntos
Terapia por Acupuntura , Transtornos Cognitivos/terapia , Cognição , Demência Vascular/terapia , Hipocampo/patologia , Transtornos da Memória/prevenção & controle , Células Piramidais/metabolismo , Pontos de Acupuntura , Animais , Astrócitos/metabolismo , Transtornos Cognitivos/etiologia , Transtornos Cognitivos/metabolismo , Transtornos Cognitivos/patologia , Demência Vascular/complicações , Demência Vascular/metabolismo , Demência Vascular/patologia , Masculino , Aprendizagem em Labirinto/fisiologia , Transtornos da Memória/etiologia , Transtornos da Memória/metabolismo , Transtornos da Memória/patologia , Ratos Wistar , Memória Espacial
14.
Pain Med ; 16(10): 1905-15, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25975413

RESUMO

BACKGROUND: According to traditional Chinese medicine (TCM) theory, acupoints are specifically chosen sites of acupuncture manipulation, and also the basis for studying the mechanism of acupuncture. Stimulating different acupoints on the body surface could provide various therapeutic benefits. However, what is the acupoint? This question is not clear. REVIEW SUMMARY: We focuse on examining the function of acupoints from different perspectives, including the local and the systemic effects of stimulating acupoints. For example, acupoints may release certain substances or incur some changes, which could adjust the function of organs, maintain homeostasis. Furthermore, the therapeutic effects of verum acupoints versus sham acupoints were discussed. However, due to insufficience in evidence and in current methodologies, research into mechanisms of acupuncture is still incomplete. CONCLUSION: This review might explain, to some extent, what an acupoint is. Further research into the identity of acupoints is warranted, and multidisciplinary methods using novel technologies may yield significant advances over existing knowledge.


Assuntos
Pontos de Acupuntura/classificação , Terapia por Acupuntura/métodos , Modelos Biológicos , Estimulação Física/métodos , Fenômenos Fisiológicos da Pele , Vísceras/fisiologia , Homeostase/fisiologia , Humanos
15.
Anal Biochem ; 400(1): 69-77, 2010 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-20035706

RESUMO

Preimplantation genetic diagnosis (PGD) is employed increasingly to allow transfer of embryos to the uterus in assisted reproduction procedures. There are three stages of biopsy: polar bodies, one or two blastomeres from the cleavage-stage embryos, and trophectoderm cells ( approximately 5cells) from the blastocyst-stage embryos. Validation of polymerase chain reaction (PCR)-based assays are challenging because only limited genetic material can be obtained for PGD. In the current study, we modified a valid single-cell PCR protocol for PGD using real-time PCR assay with fluorescence resonance energy transfer (FRET) hybridization probes followed by melting curve analysis. We optimized and clinically applied the protocol, permitting molecular genetic analysis to amplify a specific region on the beta-globin (HBB) gene for a couple, carriers of two mutations: c.-78A>G and c.52A>T. Among a total of eight embryos obtained after ovarian stimulation, a single blastomere per embryo at the six- to eight-cell stage was biopsied. This PGD method showed that four embryos were unaffected, two embryos were selected for transfer, and one pregnancy was achieved. Finally, a healthy male baby was delivered at 38weeks' gestation. The results obtained using the new method, FRET hybridization probes, were compared with findings using an existing method, primer extension minisequencing.


Assuntos
Sondas de DNA/química , Transferência Ressonante de Energia de Fluorescência , Reação em Cadeia da Polimerase/métodos , Talassemia beta/genética , Adulto , Sequência de Aminoácidos , Blastômeros/citologia , Desenvolvimento Embrionário , Feminino , Aconselhamento Genético , Humanos , Masculino , Dados de Sequência Molecular , Mutação , Gravidez , Diagnóstico Pré-Implantação , Globinas beta/genética , Talassemia beta/diagnóstico
16.
Reprod Biomed Online ; 17(5): 699-705, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18983756

RESUMO

A strategy was developed using the OmniPlex technology of whole genome amplification for preimplantation genetic diagnosis (PGD) of single gene diseases and human leukocyte antigen (HLA) haplotypes. The amplified genomic DNA library was subsequently examined separately for mutation analysis with mini-sequence and for short tandem repeat (STR) markers within the HLA loci. To evaluate the reliability of the protocol prior to PGD, tests of 50 single lymphocytes revealed an amplification efficiency of 92-96% and allele drop-out (ADO) rate of 6-16%. The strategy was validated in one beta-thalassaemia family having an affected boy. The couple underwent three cycles of ovarian stimulation and intracytoplasmic sperm injection for PGD. On 16 embryos tested, the amplification efficiency was 88-94% and ADO was 6-19%. Two cycles of embryo transfer were performed, and one pregnancy was achieved. The genotypes of the fetus were shown to be unaffected and HLA-identical, in agreement with PGD, by chorionic villus sampling. The cord blood stem cells from the newborn can be used to treat the affected sibling. This study demonstrates the first successful application of OmniPlex whole genome amplification in PGD of a single gene disorder for selecting unaffected and HLA-compatible embryos.


Assuntos
Antígenos HLA/genética , Diagnóstico Pré-Implantação/métodos , Talassemia beta/diagnóstico , Talassemia beta/genética , Adulto , Sequência de Bases , Pré-Escolar , Primers do DNA/genética , Feminino , Haplótipos , Humanos , Recém-Nascido , Masculino , Técnicas de Amplificação de Ácido Nucleico , Gravidez , Técnicas de Reprodução Assistida
17.
Bioresour Technol ; 99(6): 2072-7, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17507217

RESUMO

The objective of this study was to manufacture fire-retardant-treated low-formaldehyde-emission particleboard from recycled wood-waste particles using polymeric 4,4'-methylenediphenyl isocyanate (PMDI) and phenol-formaldehyde (PF) resins. The influence of the PMDI/PF ratio (ratio of particles sprayed with PMDI to particles sprayed with PF, w/w) after fire retardant treatment on formaldehyde emissions, mechanical properties, and surface fire resistant performance of the manufactured particleboard was investigated. The experimental results showed that the formaldehyde emissions linearly decreased with an increasing PMDI/PF ratio. Moreover, the bending strength, internal bond strength, and screw holding strength increased with an increasing PMDI/PF ratio. The thickness swelling of the particleboard was improved by using an increasing PMDI/PF ratio. Furthermore, the fire-retardant-treated low-formaldehyde-emission particleboards fabricated in our study could pass the third grade standard of surface fire resistant performance as specified by CNS 6532.


Assuntos
Biotecnologia/métodos , Formaldeído/química , Isocianatos/química , Fenóis/química , Polímeros/química , Madeira , Conservação dos Recursos Naturais , Materiais de Construção , Retardadores de Chama , Manufaturas , Teste de Materiais , Eliminação de Resíduos , Ultrassom
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