RESUMO
This study aimed to investigate the protective effect of salvinorin A on the cerebral pial artery after forebrain ischemia and explore related mechanisms. Thirty Sprague-Dawley rats received forebrain ischemia for 10 min. The dilation responses of the cerebral pial artery to hypercapnia and hypotension were assessed in rats before and 1 h after ischemia. The ischemia reperfusion (IR) control group received DMSO (1 µL/kg) immediately after ischemia. Two different doses of salvinorin A (10 and 20 µg/kg) were administered following the onset of reperfusion. The 5th, 6th, and 7th groups received salvinorin A (20 µg/kg) and LY294002 (10 µM), L-NAME (10 µM), or norbinaltorphimine (norBIN, 1 µM) after ischemia. The levels of cGMP in the cerebrospinal fluid (CSF) were also measured. The phosphorylation of AKT (p-AKT) was measured in the cerebral cortex by western blot at 24 h post-ischemia. Cell necrosis and apoptosis were examined by hematoxylin-eosin staining (HE) and TUNEL staining, respectively. The motor function of the rats was evaluated at 1, 2, and 5 days post-ischemia. The dilation responses of the cerebral pial artery were significantly impaired after ischemia and were preserved by salvinorin A treatment. In addition, salvinorin A significantly increased the levels of cGMP and p-AKT, suppressed cell necrosis and apoptosis of the cerebral cortex and improved the motor function of the rats. These effects were abolished by LY294002, L-NAME, and norBIN. Salvinorin A preserved cerebral pial artery autoregulation in response to hypercapnia and hypotension via the PI3K/AKT/cGMP pathway.
Assuntos
Isquemia Encefálica/metabolismo , Artérias Cerebrais/efeitos dos fármacos , GMP Cíclico/líquido cefalorraquidiano , Diterpenos Clerodânicos/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Pia-Máter/irrigação sanguínea , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Isquemia Encefálica/tratamento farmacológico , Artérias Cerebrais/fisiopatologia , Cromonas/administração & dosagem , GMP Cíclico/metabolismo , Modelos Animais de Doenças , Diterpenos Clerodânicos/antagonistas & inibidores , Masculino , Morfolinas/administração & dosagem , NG-Nitroarginina Metil Éster , Naltrexona/administração & dosagem , Naltrexona/análogos & derivados , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
This study aimed to investigate the effect of icariin (ICA) on thin endometrium in a rat model. To this end, 6- to 8-week-old female Sprague Dawley rats (105) were randomly divided into 7 groups: untreated, vehicle-treated (lavage with NaCl), high-dose ICA (lavage with ICA at 200 mgâkg-1âday-1), medium-dose ICA (lavage ICA at 100 mgâkg-1âday-1), low-dose ICA (lavage with ICA at 50 mgâkg-1âday-1), sham model (injected with NaCl at uterus horn), and sample group. To induce thin endometrium, rats of all groups (except sham-model) were injected with 95% ethanol via the uterine horn. Each group underwent its respective treatment for 3 estrous cycles, after which 5 rats from each group were sacrificed, and endometrial thickness was measured. The expression of CD31, factor VIII, vascular endothelial growth factor (VEGF), cytokeratin (CK), and vimentin were detected via immunohistochemistry. The results showed that CD31, factor VIII, and VEGF were primarily expressed in the cytoplasm of endometrial and vascular epithelial cells. No difference in the expression of these factors was detected between the ICA lavage groups and the untreated groups. However, high dose ICA-treated group exhibited significantly higher expression of CD31, factor VIII, and VEGF compared to that in the low dose and vehicle-treated groups. CK and vimentin in the endometrial tissue were significantly higher in the untreated and treatment groups compared to the vehicle-treated group. This study demonstrated that ICA increases thickness of the endometrium, and it may modulate expression of VEGF, CD31, and factor VIII.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Endométrio/citologia , Endométrio/efeitos dos fármacos , Flavonoides/farmacologia , Animais , Biomarcadores , Endométrio/fisiologia , Ciclo Estral , Feminino , Imuno-Histoquímica , RatosRESUMO
In the present study, we evaluated the effects of four solutions [Dulbecco's modified Eagle's medium (DMEM), sodium lactate Ringer's injection (SLRI), phosphate-buffered saline (PBS), and NaCl] on the transfection of the human protein kinase C-a antisense oligonucleotide (PKC-a ASO) aprinocarsen in human lung carcinoma A549 cells. Specifically, SLRI, DMEM, PBS, or NaCl were used as the growth solutions for A549 cells, and OPTI-MEM was used as the PKC-a ASO diluent for transfection. Additionally, SLRI, DMEM, PBS, or NaCl were used as both the growth solutions and diluents for transfection. The cell viability and transfection efficiency were determined. The results demonstrated that when SLRI was used as either the growth solution or both the growth solution and diluent for aprinocarsen transfection in A549 cells, the effects were close to the best effects observed with DMEM as the growth solution and OPTI-MEM as the diluent, which supported the transfection of aprinocarsen into the cells. Moreover, SLRI resulted in higher transfection efficiency than those of PBS and NaCl. In in vitro experiments, aprinocarsen effectively induced apoptosis in A549 cells. In conclusion, SLRI may replace PBS or NaCl in clinical trials as a transfection solution readily accepted by the human body. To our knowledge, this is the first report demonstrating the use of SLRI as a transfection solution in lung-cancer cell lines.
Assuntos
Meios de Cultura/farmacologia , Inativação Gênica/efeitos dos fármacos , Soluções Isotônicas/farmacologia , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Fosforotioatos/genética , Proteína Quinase C-alfa/antagonistas & inibidores , Cloreto de Sódio/farmacologia , Células A549 , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Humanos , Oligonucleotídeos Antissenso/metabolismo , Oligonucleotídeos Fosforotioatos/metabolismo , Proteína Quinase C-alfa/genética , Proteína Quinase C-alfa/metabolismo , Lactato de Ringer , TransfecçãoRESUMO
This study was designed to evaluate bone matrix gelatin (BMG)/fibrin glue and chitosan/gelatin composite scaffolds for cartilage tissue engineering. Chondrocytes were isolated from costal cartilage of Sprague-Dawley rats and seeded on BMG/fibrin glue or chitosan/gelatin composite scaffolds. After different in vitro culture durations, the scaffolds were subjected to hematoxylin and eosin, Masson's trichrome, and toluidine blue staining, anti-collagen II and anti-aggrecan immunohistochemistry, and scanning electronic microscopy (SEM) analysis. After 2 weeks of culture, chondrocytes were distributed evenly on the surfaces of both scaffolds. Cell numbers and the presence of extracellular matrix components were markedly increased after 8 weeks of culture, and to a greater extent on the chitosan/gelatin scaffold. The BMG/fibrin glue scaffold showed signs of degradation after 8 weeks. Immunofluorescence analysis confirmed higher levels of collagen II and aggrecan using the chitosan/gelatin scaffold. SEM revealed that the majority of cells on the surface of the BMG/fibrin glue scaffold demonstrated a round morphology, while those in the chitosan/gelatin group had a spindle-like shape, with pseudopodia. Chitosan/gelatin scaffolds appear to be superior to BMG/ fibrin glue constructs in supporting chondrocyte attachment, proliferation, and biosynthesis of cartilaginous matrix components.
Assuntos
Condrócitos/efeitos dos fármacos , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Adesivos/efeitos adversos , Agrecanas/genética , Agrecanas/metabolismo , Animais , Matriz Óssea/química , Adesão Celular , Células Cultivadas , Quitosana/efeitos adversos , Condrócitos/citologia , Condrócitos/metabolismo , Condrogênese/efeitos dos fármacos , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Fibrina/efeitos adversos , Gelatina/efeitos adversos , Ratos , Ratos Sprague-Dawley , Alicerces Teciduais/efeitos adversosRESUMO
As a heterogeneous group of disorders in pregnancy, many genetic factors are involved in the development of preeclampsia. The single nucleotide polymorphism (SNP) rs7579169, located on chromosome 2q14.2, has been shown to be associated with pregnancy-induced hypertension in Europeans. In this study, we examined whether the SNP rs7579169 is associated with the susceptibility to preeclampsia through a case-control research model in Han Chinese women. Genotypes of 145 patients with preeclampsia and 150 healthy pregnant subjects were identified by direct sequencing. The correlation between the rs7579169 genotype and the susceptibility to preeclampsia was evaluated using an unconditional logistic regression model. Although there were no differences of having the rs7579169 SNP between early onset and late onset preeclampsia, patients carrying the CT or TT genotype were more likely to develop preeclampsia than those carrying the CC genotype (CT vs CC: OR = 1.76, 95%CI = 1.07-2.87, P < 0.05; TT vs CC: OR = 5.03, 95%CI = 1.99-12.73, P < 0.05; CC vs CT + TT: OR = 2.05, 95%CI = 1.27-3.30, P < 0.05). In conclusion, although no differences of the rs7579169 SNP were identified between the early onset and late onset preeclampsia groups, we found that the CT or TT genotype and the CT+TT genotype were significantly associated with an increased risk of preeclampsia in Han Chinese women.
Assuntos
Povo Asiático/genética , Cromossomos Humanos Par 2 , Pré-Eclâmpsia/genética , Adulto , Alelos , Estudos de Casos e Controles , China , Etnicidade/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Hipertensão Induzida pela Gravidez/genética , Modelos Logísticos , Polimorfismo de Nucleotídeo Único , Gravidez , Fatores de RiscoRESUMO
Grafting influences scion photosynthetic capacity and fruit quality. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which strongly affects photosynthetic rate, and Rubisco activase (RCA), which regulates Rubisco activity, are two key photosynthetic enzymes. However, little information is available regarding the effect of grafting on the concentration and expression of Rubisco and RCA in the citrus cultivar Huangguogan. The objective of this study was to investigate the effect of grafting Huangguogan plants onto trifoliate orange, tangerine, and orange on: 1) the concentration of Rubisco and RCA; 2) the mRNA levels of rbcL, rbcS, and rca; and 3) fruit quality. Overall, the results showed that when Huangguogan plants budded on tangerine and orange, they had better fruit quality, while on trifoliate orange they had higher Rubisco concentration. Tangerine and orange are probably the most suitable rootstocks for Huangguogan plants given the environmental conditions of Sichuan Province, China.
Assuntos
Citrus/enzimologia , Produção Agrícola , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Ribulose-Bifosfato Carboxilase/genética , Citrus/genética , Citrus/fisiologia , Qualidade dos Alimentos , Frutas , Poncirus/fisiologiaRESUMO
The aim of this study was to explore the expression of PI3K, AKT, and P-AKT, and to investigate the role of PI3K/AKT signaling pathway in thin endometrium. We included 40 women treated in affiliated Shenzhen Nanshan People's Hospital of Guangdong Medical University for endometrial conditions between August 2013 and January 2015, 20 with a normal endometrium, and 20 with thin endometrium. The expression of PI3K, AKT, and P-AKT was evaluated by the immunohistochemical S-P method. The expression of PI3K, AKT, and P-AKT proteins was significantly lower in the thin endometrium group than in the normal endometrium group (P < 0.05). The expression of PI3K and AKT was positively correlated with the expression of P-AKT. The expression of PI3K, AKT, and P-AKT proteins in the thin endometrium decreases during the proliferative phase, and this process could be associated with PI3K/AKT signaling.
Assuntos
Endométrio/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfoproteínas/genética , Proteínas Proto-Oncogênicas c-akt/genética , Adulto , Estudos de Casos e Controles , Endométrio/patologia , Congêneres do Estradiol/sangue , Feminino , Regulação da Expressão Gênica , Humanos , Fosfatidilinositol 3-Quinases/sangue , Fosfoproteínas/sangue , Congêneres da Progesterona/sangue , Proteínas Proto-Oncogênicas c-akt/sangue , Transdução de SinaisRESUMO
Loquat (Eriobotrya japonica Lindl.), which originates from the cooler hill regions of southwestern China, is a typical subtropical evergreen tree. Loquat is one of the most important economic crops in China, but the available genomic information is very limited. Here, we present the first deep transcriptomic analysis of loquat. De novo assembly generated 116,723 contigs and 64,814 unigenes using Illumina sequencing technology. A total of 45,739 unigenes were annotated by Nr, GO, and COG datasets. In addition, we analyzed the gene expression profiles of loquat fruit under low temperature stress and 4017 differential expressed genes were identified. We found that the unigenes involved in the brassinosteroid biosynthesis and phosphatidylinositol signaling systems were upregulated, indicating that they have an important role in the resistance of plants to low temperature. Our results provide an invaluable resource for identification of specific genes and proteins involved in loquat development and response to low temperatures.
Assuntos
Temperatura Baixa , Eriobotrya/genética , Regulação da Expressão Gênica de Plantas , Estresse Fisiológico , Transcriptoma , Biologia Computacional , Eriobotrya/metabolismo , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Redes e Vias Metabólicas , Anotação de Sequência Molecular , Fitosteróis/metabolismo , Transdução de SinaisRESUMO
The use of noninvasive genetic sampling to identify the sex of wild animals is an extremely valuable and important tool in molecular ecology and wildlife conservation. Sex determination using the amelogenin gene has been conducted in many species because only a single pair of primers is required to amplify both X- and Y-linked alleles. However, this method has not been used in field research with the feces of wildlife. In this study, we applied this method to 222 fecal samples from wild blue sheep (Pseudois nayaur) using amelogenin primers (SE47/SE48) after testing the effectiveness of sex determination using tissue samples and fecal samples from blue sheep of known sex. We found this method to be highly reliable (80.2%) for blue sheep. Amelogenin can be used to identify the sex of wild animals using fecal samples.
Assuntos
Amelogenina/genética , Análise para Determinação do Sexo , Animais , Fezes , Análise de Sequência de DNA , Ovinos/genéticaRESUMO
We explored the effects of icariin on the expression of estrogen receptor (ER), vascular endothelial growth factor (VEGF), and kinase insert domain receptor (KDR) in the endometrial cells of the thin endometrium. Primary endometrial cells were obtained and divided into a blank control group, a high-, a middle-, and a low-dose icariin groups, as well as an estrogen treatment group to undergo cellular identification by immunocytochemistry. The expression levels of ER, VEGF, and its receptor were estimated by western blotting. The expression levels of ER, VEGF, and KDR gradually increased from the control group to the estrogen (E2) treatment and icariin treatment groups; the differences were statistically significant. However, the differences were not statistically significant among the different icariin dose groups. The endometrium may be thickened by icariin treatment by increasing the expression levels of ER, VEGF, and KDR in endometrial cells.
Assuntos
Endométrio/metabolismo , Flavonoides/farmacologia , Receptores de Estrogênio/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Adulto , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos , Endométrio/efeitos dos fármacos , Endométrio/patologia , Estradiol/farmacologia , Feminino , Humanos , Distúrbios Menstruais/tratamento farmacológico , Distúrbios Menstruais/metabolismo , Adulto JovemRESUMO
The present study aims to purify and characterize lectin from tartary buckwheat seeds and study its properties as well as biological activities to determine its possible biomedical applications in promoting maturation and proliferation of peripheral blood DCs derived from healthy donors and to study the effect of inducing apoptosis in human leukemia U937 cells. A novel tartary buckwheat lectin (TBL) protein, purified from tartary buckwheat seeds, showed a single band with a molecular mass of 65 kDa in SDS-PAGE. The purified TBL hemagglutinated both human and animal erythrocytes and showed preference for blood type O and the rabbit blood type. TBL is active at up to 60°C, and it is acid- and alkali-stable. TBL (25 µg/mL) combined with 5 x 10(-5) M rhIL-4 promotes maturation and proliferation of peripheral blood dendritic cells (DCs), which is stronger than that promoted by rhTNF-α (20 ng/mL). Exposure of DCs to 50 µg/mL TBL for 48 h resulted in extensive upregulation of maturation markers CD83 and CD40. These TBL-DCs were capable of producing several pro-inflammatory cytokines such as interleukin-10 (IL-10) and interleukin-12 (IL-12). The results of the treatment of human leukemia U937 cells with TBL in doses of 12.5, 25, 50, and 100 µg/mL showed that tartary buckwheat-derived lectin induces apoptosis in a dose-dependent manner. Our results encourage the use of tartary buckwheat and tartary buckwheat-derived lectins as immunopotentiating foods, targeted to strengthen immune responses and display a potential dietary supplement for cancer prevention.
Assuntos
Células Dendríticas/efeitos dos fármacos , Fagopyrum/química , Linfoma/patologia , Lectinas de Plantas/química , Lectinas de Plantas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Citocinas/farmacologia , Células Dendríticas/patologia , Humanos , Camundongos , Lectinas de Plantas/isolamento & purificação , Coelhos , Ratos , Sementes/química , Células U937RESUMO
In this study, five common buckwheats and nine tartary buckwheats grown at different locations were analyzed for the contents of rutin, quercetin, and amino acids by high-performance liquid chromatography and spectrophotometry. The rutin content was higher than quercetin in buckwheat seeds. Rutin content was in the range from 0.05 (0.05 g per 100 g dry seeds) to 1.35% of buckwheat seeds. Quercetin content varied from 0.01 to 0.17% and in some common buckwheats it was even difficult to detect. Comparatively, tartary buckwheat seeds contained more rutin and quercetin than common buckwheat seeds. Meanwhile, the bran has higher rutin content than the farina in tartary buckwheat seeds, with a respective content of 0.45 to 1.19% and 0.14 to 0.67%. It was found that amino acid contents were around 1.79 to 12.65% (farina) and 5.74 to 7.89% (bran) in common buckwheats, and 1.73 to 5.63% (farina) and 2.64 to 16.78% (bran) in tartary buckwheat seeds. The highest total rutin content was found to be 1.35% in tartary buckwheat seeds from Sichuan, China. The highest total amounts of amino acid were detected to be 20.13% in tartary buckwheat seeds from Changzhi, Shanxi Province (China). Our results suggested that food products made of whole-buckwheat flour are healthier than those made of fine white flour.
Assuntos
Aminoácidos/química , Fagopyrum/química , Extratos Vegetais/química , Quercetina/química , Rutina/química , Sementes/química , Aminoácidos/isolamento & purificação , China , Cromatografia Líquida de Alta Pressão , Extratos Vegetais/isolamento & purificação , Quercetina/isolamento & purificação , Rutina/isolamento & purificaçãoRESUMO
A 1794-bp cDNA fragment was amplified from mRNA isolated from pear (Pyrus pyrifolia NaKai. Cuiguan) leaves by using primers based on the sequences generated during the analysis of the pear transcriptome. The 597-amino acid sequence encoded by the cDNA was compared with the sequences in GenBank, and it was found to be similar to that of members of the sucrose-proton co-transporter family. The hydrophobic protein, which was predicted to have 11 transmembrane domains, was designated as PpSUT2. Real-time fluorescent quantitative polymerase chain reaction analysis indicated the accumulation of PpSUT2 mRNA throughout the plant, with the highest levels in the buds. Analysis of the expression of PpSUT2 during fruit development showed that the abundance of its transcripts increased at the end of April and then decreased to the lowest level at the end of July. Subcellular localization studies with the pCXDG vector as a probe demonstrated that PpSUT2 localized to cell membranes. An expression vector was constructed by inserting the PpSUT2 cDNA into pET32(a), and the vector was expressed in Escherichia coli (strain BL21) after induction with 1 mM isopropyl b-d-1-thiogalactopyranoside at 25°C. Analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis identified the induction of a 71-kDa protein. Further analysis indicated that PpSUT2 might be not directly involved in sucrose transport, instead, functioning as a sucrose sensor on the cytoplasmic membrane.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/genética , Proteínas de Plantas/genética , Pyrus/genética , Sequência de Aminoácidos , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Frutas/genética , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Proteínas de Membrana Transportadoras/química , Proteínas de Membrana Transportadoras/metabolismo , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/classificação , Proteínas de Plantas/metabolismo , Pyrus/crescimento & desenvolvimento , Pyrus/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estações do Ano , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Sacarose/metabolismoRESUMO
The diversity of microbiota in waste waters has not been thoroughly examined, despite the potential impact of microbes on effluent quality. Wastewater microbial communities harbor pathogenic bacteria, viruses, and parasites. To study microbial communities in domestic sewage outfalls, 454 pyrosequencing technology was used to investigate the composition of microbial communities associated with municipal wastewater during different seasons sampled over the course of one year. A total of 195,103 16S rRNA gene sequences were obtained from 20 samples. The R software was used to calculate the number of indices describing the alpha diversity associated with each bacterial assemblage. In this study, the a-diversity index (H', D, J), in which higher numbers represent more diversity, was found to change with seasonal cycle. The diversity of bacterial assemblages was high in all samples, indicating that species diversity was also very high. The taxonomic composition of the assemblages varied considerably among samples, with some dominated by Proteobacteria, while others were dominated by Bacteroidetes or Firmicutes. In 2 samples, the relative prevalence of Proteobacteria exceeded 90%. α-Proteobacteria, b-proteobacteria, and g-proteobacteria represented 90% or more of all Proteobacteria. The present characterization of wastewater from five sewage outfalls indicated the presence of some pathogenic bacteria. The g-Proteobacteria in sewage wastefalls identified in this study included Enterobacteriaceae, Vibrionaceae, Pseudomonadaceae, Salmonella, Yersinia, Vibrio, and Pseudomonas aeruginosa.
Assuntos
Microbiologia Ambiental , Microbiota , Esgotos/microbiologia , Bactérias/classificação , Bactérias/genética , Biodiversidade , China , Análise por Conglomerados , Geografia , Metagenoma , Filogenia , RNA Ribossômico 16SRESUMO
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
The porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a threat, exerting significant economic effects on the swine industry worldwide. However, none of the current commercially available vaccines can completely prevent respiratory infection, trans-placental transmission, pig-to-pig transmission of the virus, or maintain immune protection in sows. This study provides information on PRRSV and a review of available options for PRRS control strategies based on its pathogenic characteristics, immune properties, and biological characteristics. In this study, the nisin-controlled expression system of Lactococcus lactis was selected as a vector to express the ORF6 gene of PRRSV. Food-grade recombinant, L. lactis PNZ8149/NZ3900-M/PRRS, which contained the lactose operon, was successfully constructed. The molecular weight of the expressed recombinant protein was approximately 19 kDa. Furthermore, the recombinant protein was located on the surface of L. lactis and showed reactogenicity with the antibody against PRRSV. Results of this study are expected to lay a theoretical foundation for development of genetically engineered L. lactis mucosal vaccines and to provide information related to its immune activity and adjuvant effects.
Assuntos
Expressão Gênica/efeitos dos fármacos , Genes Virais , Lactococcus lactis/genética , Nisina/farmacologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/genética , Animais , Linhagem Celular , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Lactococcus lactis/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Suínos/virologiaRESUMO
The aim of this study was to investigate the potential correlation between polymorphisms of the estrogen receptor (ER)-ß gene (RsaI and AluI) and ethnic Han Chinese human idiopathic thin endometrium. A total of 120 patients with idiopathic thin endometrium and 120 sterility patients with normal endometrium thickness (controls) were included in the study. RsaI and AluI polymorphisms of the ER-ß gene were analyzed with polymerase chain reaction-restriction fragment length polymorphism. The distribution of polymorphisms, genotype, allele, and haplotype was compared between the 2 groups. A significant difference in the RsaI genotype was observed between idiopathic thin endometrium patients and controls. The distribution of the R allele of the RsaI polymorphism in idiopathic thin endometrium patients was 37.1% compared with 48.3% in controls. The odds ratio was 0.630 (95% confidence interval = 0.438 to 0.907, P = 0.013). No significant difference in the genotype of the AluI polymorphism was found between the two groups. The linkage disequilibrium between RsaI and AluI haploids was not significant in either group, with D' values of 0.2036 and 0.0685 in the idiopathic thin endometrium patients and controls, respectively. The results of this study suggest a potential role of the ER-ß gene polymorphisms in the etiology of idiopathic atrophic endometrium and the R allele as a potential protective factor in ethnic Han Chinese.