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Suitable biomaterials with seed cells have promising potential to repair bone defects. However, bone marrow mesenchymal stem cells (BMSCs), one of the most common seed cells used in tissue engineering, cannot differentiate efficiently and accurately into functional osteoblasts. In view of this, a new tissue engineering technique combined with BMSCs and scaffolds is a major task for bone defect repair. Lentiviruses interfering with miR-136-5p or Smurf1 expression were transfected into BMSCs. The effects of miR-136-5p or Smurf1 on the osteogenic differentiation (OD) of BMSCs were evaluated by measuring alkaline phosphatase activity and calcium deposition. Then, the targeting relationship between miR-136-5p and Smurf1 was verified by bioinformatics website analysis and dual luciferase reporter assay. Then, a rabbit femoral condyle bone defect model was established. miR-136-5p/BMSCs/ß-TCP scaffold was implanted into the defect, and the repair of the bone defect was detected by Micro-CT and HE staining. Elevating miR-136-5p-3p or suppressing Smurf1 could stimulate OD of BMSCs. miR-136-5p negatively regulated Smurf1 expression. Overexpressing Smurf1 reduced the promoting effect of miR-136-5p on the OD of BMSCs. miR-136-5p/BMSCs/ß-TCP could strengthen bone density in the defected area and accelerate bone repair. SmurF1-targeting miR-136-5p-modified BMSCs combined with 3D-printed ß-TCP scaffolds can strengthen osteogenic activity and alleviate bone defects.
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Fosfatos de Calcio , Células Madre Mesenquimatosas , MicroARNs , Osteogénesis , Impresión Tridimensional , Andamios del Tejido , Ubiquitina-Proteína Ligasas , MicroARNs/genética , MicroARNs/metabolismo , Células Madre Mesenquimatosas/metabolismo , Animales , Andamios del Tejido/química , Conejos , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Fosfatos de Calcio/química , Diferenciación Celular , Ingeniería de Tejidos/métodos , Masculino , Regeneración Ósea/genéticaRESUMEN
OBJECTIVE: To determine the developmental characteristics of flash visual evoked potentials (FVEP) and pattern-reversal visual evoked potentials (PVEP) of healthy children. METHODS: The data were collected with a Keypoint Workstation 9033A07; 168 children (2 months-13 years) were tested with FVEP and 101 (4-13 years) were tested with PVEP. RESULTS: A triphasic waveform with clear components (N2, P2, and N3) was recorded steadily after 1 year, with occurrence rates over 97% at all frequencies. FVEP latency significantly decreased with age. The amplitude difference of FVEP was greater for binocular than monocular fields. FVEP amplitude increased and amplitude differences decreased with stimulation frequency. The occurrence rate of PVEP was 100% after 4 years, and PVEP latency was significantly prolonged with age. N75 and P100 amplitudes and the N75-P100 amplitude difference increased with field of vision. CONCLUSION: FVEP can be evoked in normal children at less than 2 Hz. Stimulation frequency can be adjusted to improve early detection and verification of subclinical lesions. The PVEP waveform is simple and stable, and its results are easier to analyze and interpret than FVEP, but it is limited by visual acuity and fixation force, whereas FVEP is affected less by visual acuity. but it is necessary to establish normal reference values of each age in each laboratory because of complicated analysis. According to the specific situation of the patient (vision, fixation) and clinical demand, we need to choose the right stimulation.
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Potenciales Evocados Visuales , Humanos , Niño , Agudeza Visual , Examen Neurológico , Estimulación LuminosaRESUMEN
BACKGROUND: Nontraumatic convexal subarachnoid hemorrhage (cSAH) is a rare type of atypical subarachnoid hemorrhage. It mainly presents as a focal and transient neurological deficit with similar manifestations as transient ischemic attack. CASE SUMMARY: We report a case of a 64-year-old man who visited the hospital with paroxysmal left-sided numbness and weakness is presented in this study. Computed tomography examination indicated a high-density image of the right frontal-parietal sulcus. Digital subtraction angiography showed severe stenosis at the right anterior cerebral artery A2-A3 junction (stenosis rate approximately 70%). CONCLUSION: The findings of this case indicate that anterior cerebral artery stenosis may lead to the occurrence of cSAH.
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There are some limitations of traditional influenza vaccines concerning novel mutant strains. Therefore, it is particularly important to develop preventive means for antigen-unrelated types of influenza viruses. Recent studies have shown that probiotics can modulate the immune system and reduce the severity of viral infections. In this study, we investigated the potential of Lactiplantibacillus plantarum 0111 against influenza virus H9N2. Challenge experiments showed that L. plantarum 0111 pretreatments could effectively improve mice's survival rate and weight loss and reduce the inflammatory cytokines IL-6 and TNF-α in the lungs and bronchoalveolar lavage fluid (BALF) along with the degree of lung and intestinal injury. FMT experiment demonstrates that the protective effect produced by L. plantarum 0111 is associated with gut microorganisms. In addition, 16S high-throughput sequencing of the mouse intestinal microbiota showed that L. plantarum 0111 remodeled the intestinal microbiota after H9N2 infection and maintained the gut microbiota balance. In a mouse model, the oral administration of L. plantarum 0111 increased IFN-ß expression in the serum and BALF. At the same time, the transcript levels of IFN-ß and related ISGs in the intestine and lungs of mice were also increased. In addition, the activation and polarization of T cells in mesenteric lymph nodes (MLNs) and the spleen were detected by flow cytometry, and the results showed that L. plantarum 0111 modulated cytokines in T cells and increased IgA expression in B cells in the MLNs and spleen. Thus, L. plantarum 0111 may improve gut microbiota-mediated immune responses and thus, resist infection by the influenza virus, and it could be used as an effective preventive measure against the influenza virus.
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Gut bacterial community plays a key role in maintaining host health. The Tibetan pig (Sus scrofa), an ancient breed in China, has been known for its high adaptability to harsh environments and for its meat quality. To understand the underlying mechanisms facilitating to shape these unique features, in this study, 16S rRNA sequencing using pigs feces and subsequent bacterial functional prediction were performed. Also, the gut bacteria of two other breeds of pigs, Barkshire and Landrace, were examined for comparison. It was revealed that the structure of bacterial community in Tibetan pigs appeared to be more complex; the relative abundances of dominant bacterial families varied inversely with those of the other pigs, and the proportion of Firmicutes in Tibetan pigs was lower, but Bacteroides, Fibrobacterota, Lachnospiraceae, Oscillospiraceae, and Ruminococcaceae were higher. Bacterial functional prediction revealed that the dominant flora in the Tibetan pigs was more correlated with functions regulating the hosts' immune and inflammatory responses, such as NOD-like_receptor_signaling_pathway and vitamin metabolism. In addition, in Tibetan pigs, the taxonomic relationships in the gut bacteria on day 350 were closer than those on earlier stages. Furthermore, gender played a role in the composition and function of bacterial inhabitants in the gut; for boars, they were more correlated to drug resistance and xenobiotics metabolism of the host compared to the sows. In sum, our preliminary study on the gut bacterial composition of the Tibetan pigs provided an insight into the underlying host-microorganism interactions, emphasizing the role of intestinal bacteria in the context of modulating the host's immune system and host development.
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ARSTRACTN6-methyladenosine (m6A) methylation is the most common and abundant methylation modification of eukaryotic mRNAs, which is involved in tumor initiation and progression. The study aims to explore the potential role and the regulatory mechanism of fat mass and obesity associated (FTO) in osteosarcoma (OS) progression. In this study, we detected the expressions of Krüppel-like factor 3 (KLF3) in OS cells and tissues and found that the mRNA and protein levels of KLF3 were increased in OS cells and tissues and significantly related to tumor size, metastasis, and TNM stage and poor prognosis of OS patients. FTO promoted the proliferation and invasion and suppressed apoptosis of OS cells through cell experiments in vitro. Further mechanism dissection revealed that FTO and YTHDF2 enforced the decay of KLF3 mRNA and decreased its expression. FTO-mediated mRNA demethylation inhibited KLF3 expression in the YTHDF2-dependent manner. Moreover, KLF3 overexpression abrogated FTO-induced oncogenic effects on the proliferation and invasion of OS cells. Overall, our findings showed that FTO-mediated m6A modification of KLF3 promoted OS progression, which may provide a therapeutic target for OS.
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Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato , Neoplasias Óseas , Factores de Transcripción de Tipo Kruppel , Osteosarcoma , Adenosina/análogos & derivados , Adenosina/metabolismo , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/genética , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/metabolismo , Neoplasias Óseas/genética , Humanos , Factores de Transcripción de Tipo Kruppel/genética , Osteosarcoma/genética , ARN Mensajero/genéticaRESUMEN
The new genus and species Pulvinatusiaxuegulaensis (Brassicaceae) are described and illustrated. The species is a cushion plant collected from Xuegu La, Xizang, China. Its vegetative parts are most similar to those of Arenariabryophylla (Caryophyllaceae) co-occurring in the same region, while its leaves and fruits closely resemble those of Xerodrabapatagonica (Brassicaceae) from Patagonian Argentina and Chile. Family-level phylogenetic analyses based on both nuclear ITS and plastome revealed that it is a member of the tribe Crucihimalayeae, but the infra-/intergeneric relationships within the tribe are yet to be resolved.
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Trichinellosis is a very important food-borne parasitic disease, that seriously endangers animal husbandry and food safety. Therefore, it is necessary to develop a safe and effective vaccine against Trichinella spiralis infection. In this experiment, invasive Lactobacillus plantarum carrying the FnBPA gene served as a live bacterial vector to deliver nucleic acids to the host to produce a novel oral nucleic acid vaccine. Coexpression of the T. spiralis cathepsin F-like protease 1 gene (TsCPF1) and murine IL-4 (mIL-4) by the nucleic acid vaccine was constructed and subsequently delivered to intestinal epithelial cells via invasive L. plantarum. Thirty-seven days after the first immunization, the experimental mice were challenged with 350 T. spiralis infective larvae by oral gavage. The results showed that mice orally immune-stimulated with invasive L. plantarum pValac-TsCPF1/pSIP409-FnBPA not only produce anti-TsCPF1-specific IgG antibodies, sIgA, Th1/Th2 cytokine distinctly increased but also intestinal damage and worm burden relieved compare to non-invasive TsCPF1 group (pValac-TsCPF1/pSIP409). Most notably, experimental mice immunized with invasive L. plantarum coexpressing TsCPF1 and mIL-4 (pValac-TsCPF1-IL-4/pSIP409-FnBPA) exhibited the highest protection efficiency against T. spiralis infection. The above results reveal that invasive L. plantarum-expressing the FnBPA protein improved mucosal and cellular immunity and enhanced resistance to T. spiralis. The nucleic acid vaccine delivered by invasive L. plantarum described in this study offers a novel idea for the prevention of T. spiralis.
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Genes de Helminto , Inmunidad , Lactobacillus plantarum , Trichinella spiralis , Triquinelosis , Vacunas de ADN , Animales , Genes de Helminto/genética , Genes de Helminto/inmunología , Interleucina-4/inmunología , Lactobacillus plantarum/genética , Lactobacillus plantarum/inmunología , Ratones , Ratones Endogámicos BALB C , Trichinella spiralis/inmunología , Triquinelosis/prevención & control , Triquinelosis/veterinaria , Vacunas de ADN/administración & dosificación , Vacunas de ADN/inmunologíaRESUMEN
Understanding the interaction between the gut microbiota and Trichinella spiralis is of interest for the early diagnosis and development of therapeutics for trichinellosis and to reveal the potential role of microbiota in the mechanism of immunomodulation of this tissue-dwelling helminth. In this study, we utilized 16S rRNA gene sequencing to monitor the dynamics of the microbes in BALB/c mice challenged with T. spiralis. Flow cytometry and ELISA were used to analyze cytokines at the same time. Histopathological analysis of the duodenum was also conducted. We found that microbial perturbations occurred during infection. The abundance of the Lachnospiraceae NK4A136 group, Ruminococcus 1 and Lactococcus decreased. However, the abundance of proinflammatory Parabacteroides increased over time after infection. T. spiralis infection also tended to inhibit IFN-γ production, and promote IL-4 and IL-10 levels. In total, T. spiralis disrupts gut homeostasis and impairs the development of the intestinal ecosystem. Defining the bacterial populations affected by T. spiralis infection might help identify microbial markers for diagnosis of the disease, and the populations could also be further exploited as a novel option to treat T. spiralis infection.
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Microbioma Gastrointestinal/inmunología , Inmunidad Mucosa/inmunología , Triquinelosis/inmunología , Triquinelosis/microbiología , Animales , Ratones , Ratones Endogámicos BALB C , Trichinella spiralis/inmunologíaRESUMEN
Salmonellosis is a worldwide zoonotic disease that poses a serious threat to the reproduction of livestock and poultry and the health of young animals. Probiotics including Bacillus species, have received increasing attention as a substitute for antibiotics. In this study, chicks infected with Salmonella were fed feed supplemented with the BSH to observe the pathological changes in the liver, detect the number of viable bacteria in the liver and spleen, and record the death of the chicks. The results showed that BSH could reduce the pathological changes in the liver and the invasion of Salmonella into the liver and spleen of chicks. In addition, the survival rate of chicks in the BSH experimental group was 60%, while that in the infected control group was 26%, indicating that BSH had a protective effect on chicks infected with Salmonella. Finally, the fecal microflora of 9-day-old chicks was analyzed by 16S rRNA high-throughput sequencing. The results showed that Salmonella infection could cause intestinal flora changes, while BSH could alleviate this change. In addition, BSH also promoted the proliferation of Lactobacillus salivarius in the cecum of chick. This study emphasized that BSH has anti- Salmonella infection effects in chickens and can be used as a candidate microecological preparation strain.
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Microbioma Gastrointestinal , Enfermedades de las Aves de Corral , Probióticos , Salmonelosis Animal , Alimentación Animal , Animales , Bacillus subtilis , Ciego , Pollos , Enfermedades de las Aves de Corral/prevención & control , ARN Ribosómico 16S/genética , Salmonelosis Animal/prevención & controlRESUMEN
BACKGROUND: The epidemiology on the human papillomavirus (HPV) among females in Southern China is not well-established. Baseline data on the prevalence of HPV infection in China prior to mass prophylactic HPV vaccination would be useful. Thus, this study aims to determine the type-specific HPV prevalence and distribution among females from Southern China prior to mass HPV vaccination. METHODS: A retrospective cross-sectional study employing 214,715 women attending ChenZhou NO.1 People's Hospital for cervical screening during 2012-2018 was conducted prior to widespread HPV vaccination. HPV genotype was detected using nucleic acid molecular diversion hybridization tests. The overall prevalence, age-specific prevalence, type distribution, and annual trend were analyzed. RESULTS: The overall HPV prevalence was 18.71% (95% confidence interval [CI], 18.55-18.88%) among Southern China females. During 2012-2018, the prevalence of HPV infection showed a downward tendency, from 21.63% (95% CI, 21.07-22.20%) in 2012 to 18.75% (95% CI, 18.35-19.16%) in 2018. Age-specific HPV distribution displayed a peak at young women aged less than 21 years (33.11, 95% CI, 31.13-35.15%), 20.07% (95% CI, 19.70-20.44%) among women aged 21-30 years, 17.29% (95% CI, 17.01-17.57%) among women aged 31-40 years, 17.23% (95% CI, 16.95-17.51%) among women aged 41-50 years, 21.65% (95% CI, 21.11-22.20%) among women aged 51-60 years, and 25.95% (95% CI, 24.86-27.07%) among women aged over 60 years. Of the 21 subtypes identified, the top three prevalent high-risk HPV (HR-HPV) genotypes were HPV52 (5.12%; 95% CI, 21.11-22.20%), - 16 (2.96%; 95% CI, 2.89-3.03%), and - 58 (2.51%; 95% CI, 2.44-2.58%); the predominant low-risk HPV (LR-HPV) genotypes were HPV81 (1.86%; 95%CI, 1.80-1.92%) and - 6 (0.69%; 95% CI, 0.66-0.73%) respectively. Incidence of HR-HPV only, LR-HPV only and mixed LR- and HR-HPV were 15.17, 2.07 and 1.47% respectively. Besides, single HPV infection accounted for 77.30% of all positive cases in this study. CONCLUSIONS: This study highlights 1) a high prevalence of HPV infection among females with a decreasing tendency towards 2012-2018, especially for young women under the age of 21 prior to mass HPV vaccination; 2) HPV52, - 16 and - 58 were the predominant HPV genotypes, suggesting potential use of HPV vaccine covering these HPV genotypes in Southern China.
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Alphapapillomavirus/genética , Infecciones por Papillomavirus/epidemiología , Infecciones por Papillomavirus/virología , Adolescente , Adulto , China/epidemiología , Estudios Transversales , Femenino , Genoma Viral , Humanos , Vacunación Masiva , Persona de Mediana Edad , Vacunas contra Papillomavirus/administración & dosificación , Prevalencia , Estudios Retrospectivos , Adulto JovenRESUMEN
BACKGROUND: Eimeria spp. are responsible for chicken coccidiosis which is the most important enteric protozoan disease resulting in tremendous economic losses in the poultry industry. Understanding the interaction between the avian cecal microbiota and coccidia is of interest in the development of alternative treatments that do not rely on chemotherapeutics and do not lead to drug resistance. METHODS: We utilized 16S rRNA gene sequencing to detect the dynamics of the cecal microbial community in AA broilers challenged with Eimeria tenella. Histopathological analysis of the cecum was also conducted. RESULTS: We found that microbial shifts occur during the infection. Lactobacillus, Faecalibacterium, Ruminococcaceae UCG-013, Romboutsia and Shuttleworthia decreased in abundance. However, the opportunistic pathogens Enterococcus and Streptococcus increased in abundance over time in response to the infection. CONCLUSIONS: Eimeria tenella disrupts the integrity of the cecal microbiota and could promote the establishment and growth of potentially pathogenic bacteria. Defining bacterial populations affected by coccidial infection might help identify bacterial markers for intestinal disease as well as populations or species that could be beneficial in maintaining and restoring gut homeostasis during and after infection with E. tenella.
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Pollos , Coccidiosis/veterinaria , Eimeria tenella , Microbioma Gastrointestinal/genética , Enfermedades de las Aves de Corral/parasitología , Animales , Bacterias/clasificación , Bacterias/aislamiento & purificación , Ciego/microbiología , Ciego/parasitología , Ciego/patología , Pollos/microbiología , Pollos/parasitología , Coccidiosis/terapia , Eimeria tenella/genética , Eimeria tenella/parasitología , Metagenómica , ARN Ribosómico 16S/genéticaRESUMEN
Persistent infection with the carcinogenic human papillomavirus (HPV) is a prerequisite for the progression of cervical lesions and cancer. A growing body of research has focused on the functional role of the vaginal microbiota in the persistence of HPV infection. Understanding the microbial composition and structure in women with high-risk (hr)-HPV infection may help reveal associations between the vaginal microbiota and HPV infection, and identify potential biomarkers. Our study investigated the vaginal microbial community in women with and without hr-HPV infection, by using 16s rRNA gene sequencing. We found that microbial perturbations occurred in the early phase of hr-HPV infection. Lactobacillus and Sporolactobacillus were decreased, while bacteria related to bacterial vaginosis (BV), such as Gardnerella, Prevotella, Dialister, Slackia, Actinomyces, Porphyromonas, Peptoniphilus, Anaerococcus, Peptostreptococcus, Streptococcus, Ureaplasma, Megasphaera, and Mycoplasma were increased. Our results could offer insights into the correlations between hr-HPV and the vaginal microbiota in the early infection period, and provide indications that the predominance of some BV-associated bacteria during hr-HPV infection may increase the risk for cervical neoplasia.
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Microbiota , Infecciones por Papillomavirus , Femenino , Humanos , Microbiota/genética , Papillomaviridae/genética , Infecciones por Papillomavirus/epidemiología , ARN Ribosómico 16S/genética , VaginaRESUMEN
PURPOSE: Only a few studies have investigated the brain morphology abnormalities in structural MRI in patients with drug-naïve idiopathic generalized epilepsy (IGE) and mainly focused on brain volume changes. In the present study, we aimed to investigate the changes in three morphologic measurement differences including cortical thickness, cortical volume, and surface area using FreeSurfer in a pediatric cohort of recent-onset, drug-naïve IGE. METHODS: Forty-five recent-onset, drug-naïve patients diagnosed with IGE and 32 demographically matched healthy controls were recruited. All participants underwent structural MRI scans with a 3.0â¯T MR system. FreeSurfer, an automated cortical surface reconstruction toolbox, was applied to compare the cortical morphology between patients and controls. The brain regions with significant group differences after multiple comparison correction were extracted in common space for each patient, and then correlated with their clinical characteristics (including onset age, duration of epilepsy, and mini-mental state examination (MMSE)) using partial correlation analysis with age, sex and intracranial volume as covariates. RESULTS: Compared with controls, IGE patients showed decreased cortical thickness in the left rostral middle frontal gyrus, decreased cortical volume in the right cuneus and left superior frontal gyrus that extended to the precentral gyrus, and decreased surface area in the right cuneus and right inferior parietal gyrus. None of these regions showed significant relationships with clinical measurements in the patient group. CONCLUSION: Our findings suggest that cortical thickness, cortical volume, and surface area changes occurred in the early stage of IGE. These findings provide structural neuroimaging evidence underlying the pathology of IGE.
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Mapeo Encefálico , Epilepsia Generalizada/diagnóstico por imagen , Lóbulo Frontal/diagnóstico por imagen , Imagen por Resonancia Magnética , Corteza Prefrontal/diagnóstico por imagen , Adolescente , Adulto , Estudios de Casos y Controles , Niño , Preescolar , Epilepsia/diagnóstico por imagen , Femenino , Lóbulo Frontal/patología , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Masculino , Reconocimiento de Normas Patrones Automatizadas , Corteza Prefrontal/patología , Reproducibilidad de los ResultadosRESUMEN
OBJECTIVES: This study aimed to determine the relationship between the polymorphisms of the H1/H2 gene of platelet membrane receptor P2Y12 and cerebral infarction (CI) in a Han population in North Shandong Province, People's Republic of China. PATIENTS AND METHODS: A case-control study, which involved 168 nonstoke subjects (contrast group) and 152 CI patients (CI group), was conducted. The state of subjects in the CI group was validated by computed tomography or MRI. The clinical data were categorized into two groups. The data included age, gender, smoking, drinking, shrinkage pressure, diastolic blood pressure, blood glucose, cholesterol, triglyceride, low-density lipoprotein, high-density lipoprotein, serum uric acid, fibrinogen and homocysteine. The polymorphisms were genotyped with PCR and restriction fragment length polymorphism analysis. The distribution characteristics of nonstoke subjects and CI patients and the relationship between the polymorphisms of the H1/H2 gene of platelet membrane receptor P2Y12 and ischemic stroke were analyzed. RESULTS: No significant difference was found between the contrast group and CI group (P>0.05) in terms of age, gender composition, smoking, alcohol consumption, blood glucose, cholesterol, triglyceride, low-density protein, high-density lipoprotein cholesterol, uric acid and homocysteine. In contrast, significant differences were found between these two groups (P<0.01) in terms of SBP, DBP and plasma fibrinogen level. The genotyping revealed 112 carriers of the wild-type H1/H1 genotype and 40 carriers of the mutational H2 allele of P2Y12 H1/H2 in the CI group and 140 carriers of the wild-type H1/H1 genotype and 28 carriers of the mutational H2 allele of P2Y12 H1/H2 in the contrast group. Furthermore, the H1/H2 and H2/H2 gene frequencies (26.3%) were significantly higher in the CI group (χ2=4.440, P<0.05) than those in the contrast group (16.7%). Moreover, the frequencies of the H2 allele in the CI and contrast groups were 14.5% and 8.6%, respectively, and the difference was statistically significant (χ2=5.392, P<0.05). Multiple logistic regression analysis results revealed that factors associated with CI include systolic blood pressure and plasma fibrinogen level, which carry the -893T gene. After adjusting for potential confounding factors, the H2 allele carriers had a 1.928-fold increased risk for CI (OR=1.928, 95% confidence interval: 1.137-3.188; P=0.038) when compared with noncarriers. CONCLUSION: The present study found that hypertension and elevated plasma fibrinogen levels are significant risk factors for ischemic stroke and confirmed that the H1/H2 and H2/H2 genes of platelet membrane glycoprotein receptor P2Y12 are risk factors of ischemic stroke.
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Porcine epidemic diarrhea virus (PEDV) is one of the most important causative pathogens of swine diarrhea, which is widely prevalent throughout the world and is responsible for significant economic losses in the commercial pig industry, both domestic and abroad. The spike (S) protein in the PEDV capsid structure can carry the major B lymphocyte epitope, which induces production of neutralizing antibodies and provides immunoprotective effects. Moreover, the conserved region encoded by the S gene can be considered a target for establishing a new diagnostic method and is a new candidate for vaccine design. In this study, use of anchorin pgsA' allowed the fusion protein of S-DCpep to express on the surface of recombinant Lactobacillus plantarum (NC8-pSIP409-pgsA'-S-DCpep) NC8 strain. Mice were immunized by lavage administration of the recombinant NC8-pSIP409-pgsA'-S-DCpep, which was observed to induce DC activation and high production of sIgA and IgG antibodies in experimental animals, while also eliciting production of significantly more IgA+B220+ B cells. More importantly, secretion of cytokines IFN-γ, IL-4 and IL-17 in mice that were vaccinated with NC8-pSIP409-pgsA'-S-DCpep was remarkably increased. The results of our study suggest that NC8-pSIP409-pgsA'-S-DCpep potently triggers cellular and humoral immune responses. The obtained experimental results can provide a theoretical basis that lays the foundation for production of a novel oral vaccine against PED.
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Anticuerpos Antivirales/biosíntesis , Lactobacillus plantarum/genética , Péptidos/inmunología , Virus de la Diarrea Epidémica Porcina/inmunología , Glicoproteína de la Espiga del Coronavirus/inmunología , Vacunas Virales/administración & dosificación , Administración Oral , Animales , Linfocitos B/inmunología , Linfocitos B/virología , Proliferación Celular , Femenino , Expresión Génica , Inmunogenicidad Vacunal , Inmunoglobulina A/biosíntesis , Inmunoglobulina G/biosíntesis , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-17/genética , Interleucina-17/inmunología , Interleucina-4/genética , Interleucina-4/inmunología , Lactobacillus plantarum/metabolismo , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Péptidos/genética , Virus de la Diarrea Epidémica Porcina/genética , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Glicoproteína de la Espiga del Coronavirus/genética , Vacunación , Vacunas Virales/biosíntesis , Vacunas Virales/genéticaRESUMEN
BACKGROUND: Chlamydia pneumoniae (C. pneumoniae) is pathogenic to humans, by causing pulmonary inflammation or bronchitis in both adolescents and young adults. However, the molecular signals linking C. pneumoniae components to inflammation remain elusive. This study was to investigate the effect of Chlamydia-specific Cpn0423 of C. pneumoniae on C. pneumoniae-mediated inflammation. RESULTS: Cpn0423 was detected outside of C. pneumoniae inclusions, which induced production of several cytokines including macrophage inflammatory protein-2 (MIP-2) and interleukins (ILs). Production of the Cpn0423-induced cytokines was markedly reduced in cells pretreated with NOD2-siRNA, but not with negative control oligonucleotides. Mice treated with Cpn0423 through intranasal administration exhibited pulmonary inflammation as evidenced by infiltration of inflammatory cells, increased inflammatory scores in the lung histology, recruitment of neutrophils and increased cytokines levels in the BALF. CONCLUSION: Cpn0423 could be sensed by NOD2, which was identified as an essential element in a pathway contributing to the development of C. pneumoniae -mediated inflammation.
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Proteínas Bacterianas/inmunología , Infecciones por Chlamydophila/inmunología , Chlamydophila pneumoniae/inmunología , Mediadores de Inflamación/inmunología , Proteína Adaptadora de Señalización NOD2/inmunología , Neumonía Bacteriana/microbiología , Animales , Proteínas Bacterianas/genética , Quimiocina CXCL2/genética , Quimiocina CXCL2/inmunología , Infecciones por Chlamydophila/genética , Infecciones por Chlamydophila/microbiología , Chlamydophila pneumoniae/genética , Humanos , Interleucinas/genética , Interleucinas/inmunología , Pulmón/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína Adaptadora de Señalización NOD2/genética , Neumonía Bacteriana/genética , Neumonía Bacteriana/inmunologíaRESUMEN
Recombination activating gene 2 (RAG2) is necessary for immature B cell differentiation. Antibodies to human and rabbit RAG2 are currently commercially available, but antibodies to swine RAG remain unavailable to date. In this study, the swine RAG2 genes sequence was synthesized and then cloned into a pET-28a vector. The recombinant fusion protein was successfully expressed in E. coli, purified through nickel column chromatography, and further digested with Tobacco Etch Virus protease. The cleaved protein was purified by molecular-exclusion chromatography and named pRAG2. We used pRAG2 to immunize rabbits, collected the serum and purified rabbit anti-pRAG2 polyclonal antibodies. The rabbit anti-pRAG2 polyclonal antibodies were tested via immunofluorescence on eukaryotic cells overexpressing pRAG2 and also able to recognize pig natural RAG2 and human RAG2 protein in western blotting. These results indicated that the prepared rabbit anti-pRAG2 polyclonal antibodies may serve as a tool to detect immature B cell differentiation of swine.
Asunto(s)
Anticuerpos/química , Proteínas de Unión al ADN/biosíntesis , Escherichia coli/genética , Expresión Génica , Proteínas Nucleares/biosíntesis , VDJ Recombinasas/biosíntesis , Animales , Anticuerpos/aislamiento & purificación , Anticuerpos/metabolismo , Western Blotting , Clonación Molecular , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Endopeptidasas/química , Escherichia coli/metabolismo , Técnica del Anticuerpo Fluorescente , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Humanos , Sueros Inmunes/química , Isoenzimas/biosíntesis , Isoenzimas/genética , Isoenzimas/inmunología , Proteínas Nucleares/genética , Proteínas Nucleares/inmunología , Conejos , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Porcinos , VDJ Recombinasas/genética , VDJ Recombinasas/inmunologíaRESUMEN
PURPOSE: To evaluate the prognostic effect of central obesity on triple negative breast cancer (TNBC). METHODS: 206 TNBC patients treated from June 2006 to June 2015 were enrolled retrospectively. Body mass index (BMI) ≥25 kg/m(2) was the standard of obesity and waist circumference ≥80 cm was the standard of central obesity. Patient and tumor characteristics were compared between obesity categories. Survival differences between obesity categories were assessed with log-rank test in the univariate analysis and prognostic factors were then investigated by Cox regression analysis. RESULTS: 81 cases were with obesity (39.3 %). 71 cases were with central obesity (34.5 %). Patients with obesity or central obesity tended to be older (P = 0.022 for obesity; P = 0.013 for central obesity) and to have larger tumor size (P = 0.027 for obesity; P = 0.027 for central obesity). By Cox regression analysis, central obesity (DFS: HR 1.759; 95 % CI 1.009-3.065; P = 0.046. OS: HR 2.297; 95 % CI 1.184-4.456; P = 0.014) was identified as an independent prognostic factor. For central obesity with BMI ≥ 25 kg/m(2), the prognostic effect was more apparent (DFS: HR 1.845; 95 % CI 1.059-3.212; P = 0.031. OS: HR 2.377; 95 % CI 1.230-4.593; P = 0.010). CONCLUSION: Central obesity, especially with high BMI, was an independent prognostic factor for TNBC.
RESUMEN
During a study of ornithophilous viruses in China, a new densovirus (DNV) was isolated from the lung tissue of Parus major (PmDNV-JL). The complete genome of PmDNV-JL was cloned and sequenced. Five open reading frames (ORFs) were identified in the 5166nt sequence, on the basis of deduced amino acids. It was further shown that this virus caused cytopathic effects (CPE) in Feline kidney cells. The NS1 gene sequence of PmDNV-JL shares 70-99% nucleotide sequence identity with isolates of the Blattella germanica densovirus (BgDNV) and BgDNV-like virus. Phylogenetic analysis indicated that the predicted amino acid sequences of capsid (VP) and non-structural domain (NS1) of PmDNV-JL clustered with the BgDNV and were similar to BgDNV-HB within the genus Densovirus.