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Head and neck squamous cell carcinoma is the sixth most common cancer worldwide and has high heterogeneity and unsatisfactory outcomes. To better characterize the tumor progression trajectory, we perform single-cell RNA sequencing of normal tissue, precancerous tissue, early-stage, advanced-stage cancer tissue, lymph node, and recurrent tumors tissue samples. We identify the transcriptional development trajectory of malignant epithelial cells and a tumorigenic epithelial subcluster regulated by TFDP1. Furthermore, we find that the infiltration of POSTN+ fibroblasts and SPP1+ macrophages gradually increases with tumor progression; their interaction or interaction with malignant cells also gradually increase to shape the desmoplastic microenvironment and reprogram malignant cells to promote tumor progression. Additionally, we demonstrate that during lymph node metastasis, exhausted CD8+ T cells with high CXCL13 expression strongly interact with tumor cells to acquire more aggressive phenotypes of extranodal expansion. Finally, we delineate the distinct features of malignant epithelial cells in primary and recurrent tumors, providing a theoretical foundation for the precise selection of targeted therapy for tumors at different stages. In summary, the current study offers a comprehensive landscape and deep insight into epithelial and microenvironmental reprogramming throughout initiation, progression, lymph node metastasis and recurrence of head and neck squamous cell carcinoma.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Humanos , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Carcinoma de Células Escamosas/patología , Metástasis Linfática , Linfocitos T CD8-positivos/metabolismo , Ecosistema , Recurrencia Local de Neoplasia/genética , Neoplasias de Cabeza y Cuello/genética , Microambiente Tumoral/genéticaRESUMEN
Objective: To explore the classification and reconstruction strategy of defects in lateral face region after operation of malignant tumors. Methods: Eighteen cases with the reconstruction of complicated defects after resection of tumors in the region of lateral face from January 2015 to January 2018 in Hunan Cancer Hospital were retrospectively reviewed. There were 14 males and 4 females, aged from 32 to 68 years. According to the presence or absence of bony scaffold, complicated defects were divided into two main categories: soft tissue perforating defects and soft tissue defects combined with bony scaffold defects. All soft tissue perforating defects in 5 cases were repaired with free anterolateral femoral flaps. Among 13 cases with soft tissue plus bony scaffold defects, 3 were repaired with free fibular flaps, 6 with free fibular flaps combined with free anterolateral femoral flaps, and 4 with chimeric deep circumflex iliac artery perforator flaps combined with anterolateral femoral flaps. Results: All flaps survived well. Two patients complicated with fistula in floor of mouth, but the wound healed after dressing change. Transoral feeding was resumed within 2 weeks after surgery in all patients. One year follow-up evaluation showed that 14 cases had symmetrical face and 10 cases had mouth opening more than 3 transverse fingers. After 36-50 months of follow-up, 6 patients died, with an overall 3-year survival rate of 66.7%. Conclusion: The classification of defects with or without bony stent loss is conducive to the overall repair design, the recovery of facial contour stent, the effective fill of dead space and the maintain of residual occlusal relationship. Good reconstruction results require a multi flap combination of osteocutaneous and soft tissue flaps.
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Neoplasias Faciales , Colgajos Tisulares Libres , Colgajo Perforante , Procedimientos de Cirugía Plástica , Traumatismos de los Tejidos Blandos , Femenino , Humanos , Masculino , Estudios Retrospectivos , Trasplante de Piel , Traumatismos de los Tejidos Blandos/cirugía , Muslo/cirugíaRESUMEN
OBJECTIVE: To evaluate the effects of femtosecond laser treated microgrooved surface on microscopic topography, phase transformation, and three-points flexural strength of zirconia, and to provide reference for surface microstructure optimization of zirconia implant. METHODS: According to different surface treatment methods, 57 computer aided design/computer aided manufacture (CAD/CAM) zirconia bars (20.0 mm×4.0 mm×1.4 mm) were evenly divided into three groups: sintered group, no treatment after sintering, taken as control; sandblasted group, sandblasted with 110 µm aluminium oxide (Al2O3) after sintering; microgrooved group, femtosecond laser fabricated microgrooves with 50 µm width, 30 µm depth, and 100 µm pitch. Surface microscopic topography was observed with scanning electron microscope (SEM) and 3D laser microscope. Further, surface roughness in each group and microgroove size were measured. Crystal phase was analyzed with X-ray diffraction. Specimens were subjected to three- points flexural strength test, and Weibull distribution was used to analyze their strength characteristics. RESULTS: SEM showed that sintered surface was flat with clear grain structure; sandblasted surface exihibited bumps and holes with sharp margins and irregular shape; microgrooves were regularly aligned without evident defect, and nano-scale particles were observed on the surface inside of the microgrooves. Ra value of microgrooved group [(9.42±0.28)] µm was significantly higher than that of sandblasted group [(1.04±0.03) µm] and sintered group [(0.60±0.04) µm], and there was statistical difference between sandblasted group and sintered group (P < 0.001). The microgroove size was precise with (49.75±1.24) µm width, (30.85±1.02) µm depth, and (100.58±1.94) µm pitch. Crystal phase analysis showed that monoclinic volume fraction of sandblasted group (18.17%) was much higher than that of sintered group (1.55%), while microgrooved group (2.21%) was similar with sintered group. The flexural strength of sandblasted group (986.22±163.25) MPa had no statistical difference with that of sintered group (946.46±134.15) MPa (P=0.847), but the strength in microgrooved group (547.92±30.89) MPa dropped significantly compared with the other two groups (P < 0.001). Weibull modulus of sintered, sandblasted, microgrooved groups were 7.89, 6.98, and 23.46, respectively. CONCLUSION: Femtosecond laser was able to form micro/nanostructured microgrooves on zirconia surface, which deleteriously affected the flexural strength of zirconia.
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Cerámica , Resistencia Flexional , Materiales Dentales , Humanos , Rayos Láser , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Propiedades de Superficie , Itrio , CirconioRESUMEN
Objective: To understand the current status of HIV self-testing reagent use in pre-exposure prophylaxis (PrEP) and related factors in men who have sex with men (MSM). Methods: From December 2018 to December 2019, "Gold data" online platform (www.jinshuju.com) was used to conduct multicenter PrEP studies in Shenyang, Beijing, Chongqing and Shenzhen of China. Results: A total of 1 222 MSM PrEP users were included in the multicenter study. The average age of the participants was (31.5±8.7) years, and the number of sexual partners in the past three months was 3 (P25,P75:2,6). The proportions of those who did not use condoms in anal sex with fixed, casual and commercial partners were 62.7% (456/727), 56.3% (440/781) and 41.0% (16/39), respectively. Up to 74.5% (910/1 222) of participants had used HIV self-testing reagents, and the number of HIV self-testing during last year was 3 (P25,P75:2,5). The multivariate logistic regression analysis indicated that compared with age group >40 years, those with education level of junior high school or below, those with psychological identity as female, event driven PrEP users, those never using new type drugs in past 3 months, the participants aged 18- years (aOR=2.06, 95%CI: 1.35-3.14), 26- years (aOR=2.72, 95%CI: 1.77-4.17), 31- years (aOR=1.76, 95%CI: 1.19-2.59), undergraduates (aOR=2.18, 95%CI: 1.35-3.49), graduate students and above (aOR=3.06, 95%CI: 1.69-5.54), those with psychological identity as male (aOR=3.22, 95%CI: 1.55-6.70), daily PrEP users (aOR=1.35, 95%CI: 1.03-1.78), and new type drug users in the past three months (aOR=1.72, 95%CI: 1.30-2.28) had higher proportions of HIV self-testing behaviors. Conclusions: The proportion of HIV self-testing in MSM PrEP users was high, while it was relatively low in older age group, event driven PrEP users and MSM never using new type drugs. To assess and improve the effectiveness and compliance of PrEPs, it is necessary to provide better HIV self-testing service for MSM with low HIV self-testing rate.
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Infecciones por VIH , Homosexualidad Masculina , Profilaxis Pre-Exposición , Autoevaluación , Adolescente , Adulto , China , Infecciones por VIH/prevención & control , Homosexualidad Masculina/estadística & datos numéricos , Humanos , Indicadores y Reactivos , Masculino , Factores Socioeconómicos , Adulto JovenRESUMEN
Objective: To evaluate the clinical characteristics and prognosis of gallbladder cancer (GBC) patients in China. Methods: This retrospective multicenter cohort study enrolled 3 528 consecutive GBC patients diagnosed between January 2010 to December 2017 in 15 hospitals from 10 provinces. There were 1 345 (38.12%) males and 2 183 (61.88%) females.The age of diagnosis was (63.7±10.8) years old (range: 26 to 99 years old) .There were 213 patients (6.04%) in stage 0 to â , whereas 1 059 (30.02%) in stage â ¡ to â ¢, 1 874 (53.12%) in stage â £, and 382 (10.83%) unavailable. Surgery was performed on 2 255 patients (63.92%) . Three hundred and thirty-six patients received chemotherapy or radiotherapy (9.52%; of which 172 were palliative); 1 101 (31.21%) received only supportive treatment.The patient source, treatment and surgery, pathology, concomitant gallstone, and prognosis were analyzed. Results: Among the 3 528 GBC patients, 959 (27.18%) were from East China, 603 (17.09%) from East-North China, 1 533 (43.45%) from Central China, and 433(12.27%) from West China. Among the 1 578 resectable tumor, 665 (42.14%) underwent radical surgery, 913 (57.86%) underwent surgery that failed to follow the guidelines.Eight hundred and ninety-one (56.46%) patients were diagnosed before surgery, 254 (16.10%) during surgery, and 381 (24.14%) after surgery (time point of diagnosis couldn't be determined in 52 patients) .Among the 1 578 patients with resectable tumor, 759 (48.10%) had concomitant gallstone.Among the 665 patients underwent radical surgery, 69 (10.4%) showed positive resection margin, 510 (76.7%) showed negative resection margin, and 86 (12.9%) unreported margin status.The 5-year overall survival rate (5yOS) for the 3 528-patient cohort was 23.0%.The 5yOS for patients with resectable tumor was 39.6%, for patients with stage â £B tumor without surgery was 5.4%, and for patients with stage â £B tumor underwent palliative surgery was 4.7%. Conclusions: More than half GBC patients in China are diagnosed in stage â £.Curative intent surgery is valuable in improving prognosis of resectable GBC.The treatment of GBC needs further standardization.Effective comprehensive treatment for GBC is in urgent need.
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Neoplasias de la Vesícula Biliar/terapia , Adulto , Anciano , Anciano de 80 o más Años , China , Femenino , Neoplasias de la Vesícula Biliar/patología , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Pronóstico , Estudios RetrospectivosRESUMEN
Five patients with primary malignancies in the pterygopalatine fossa (PPF) and infra temporal fossa (ITF) were enrolled in this retrospective study between January 2012 and January 2018. After malignancies proven by biopsy and evaluation with CT and MRI scan, all patients received modified maxillary swing (MMS) approach for extirpation of malignant tumors in the PPF and ITF under general anesthesia. En bloc resection with wide surgical margins was successfully performed in all cases. Negative margins were observed in 4 cases and positive margins were found in one patient with adenoid cystic carcinoma who received postoperative radiotherapy. The most common complication was facial numbness. During the follow-up period (range 12 to 57 months), one patient suffered from recurrence while others did not. The advantages of MMS include wide surgical field, full exposure and easy manipulation. The MMS approach is expected to become an standard method for monobloc resection of malignancies in the PPF and ITF.
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Neoplasias de la Base del Cráneo , Humanos , Recurrencia Local de Neoplasia , Fosa Pterigopalatina , Estudios Retrospectivos , Neoplasias de la Base del Cráneo/cirugíaRESUMEN
Objective: To compare the efficacy and safety of mycophenolate mofetil versus cyclosporine A in treating children with primary refractory nephrotic syndrome. Methods: Conducted a prospective randomized controlled clinical trial in 62 pediatric patients (including 44 boys and 18 girls), age ranged from 2.1 to 17.0 years; 32 cases presented with frequently relapsing nephrotic syndrome (FRNS) and 30 cases presented with steroid-resistant nephrotic syndrome (SRNS), who were admitted to department of Nephrology, Children's Hospital Affiliated to Capital Institute of Pediatrics from October 2013 to October 2015. The patients received either mycophenolate mofetil (20-30)mg/(kg·d) or cyclosporine A (3-5)mg/(kg·d) randomly, on the basis of prednisone treatment. Follow-up interview was conducted regularly for at least one year. Efficacy rate, relapse rate, time required for induction of remission, relapse-free period and prednisone dosage were compared between the two groups. Results: (1) Renal histologic examination, which was available for 17 patients, revealed minimal change disease in 8 patients, mesangial proliferative glomerulonephritis (MsPGN) in five, membranous nephropathy in two, and focal segmental glomerulosclerosis (FSGS) in two. (2) Comparison of mycophenolate mofetil versus cyclosporine A in children with FRNS: There were 14 patients with FRNS in mycophenolate mofetil group and 18 patients with FRNS in cyclosporine A group respectively. The relapse rate (episodes/year) in cyclosporine A group was lower than that of mycophenolate mofetil group (1.0 (0.0, 1.0) vs. 1.0 (1.0, 3.0), Z=-2.405, P=0.016). The relapse-free period (months) in cyclosporine A group was longer than that of mycophenolate mofetil group (10.0 (5.7, 12.1) vs. 5.0 (1.0, 11.0), Z=-1.984, P=0.047). No significant difference in dosage of prednisone was found between cyclosporine A and mycophenolate mofetil groups when followed up for 1 year. (3) Comparison of mycophenolate mofetil versus cyclosporine A in children with SRNS: The efficacy rate was 6/14 in mycophenolate mofetil group and 13/16 in cyclosporine A group. The complete remission rate was 4/14 in mycophenolate mofetil group and 12/16 in cyclosporine A group (P<0.05). The time (months) required for induction of remission in cyclosporine A group was significantly shorter than that of mycophenolate mofetil group (1.0 (1.0, 2.0) vs. 3.0 (2.5, 4.0), Z=-2.529, P=0.011). No significant differences were found between the two groups with respect to relapse-free period and relapse rate. (4) Except that one patient developed hypertensive encephalopathy in cyclosporine A group, no other serious adverse events were recorded. There were no significant differences between two groups with respect to adverse events. Conclusion: Our results indicated that both mycophenolate mofetil and cyclosporine A were effective in the treatment of children with refractory nephrotic syndrome. Cyclosporine A was superior to mycophenolate mofetil in preventing relapses in patients with FRNS and inducing complete remission in patients with SRNS. Although most patients were able to tolerate mycophenolate mofetil and cyclosporine A, but the toxicity and safety of cyclosporine A should be monitored closely.
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Ciclosporina , Inmunosupresores , Ácido Micofenólico , Síndrome Nefrótico , Adolescente , Niño , Preescolar , Ciclosporina/uso terapéutico , Quimioterapia Combinada , Femenino , Humanos , Inmunosupresores/uso terapéutico , Masculino , Ácido Micofenólico/uso terapéutico , Síndrome Nefrótico/tratamiento farmacológico , Estudios Prospectivos , Resultado del TratamientoRESUMEN
Recently, the one-stage posterior approach for treating spinal tuberculosis (TB) has gained popularity. However, large bony defects after debridement remain a major challenge in posterior surgery. The present retrospective study aims to compare the clinical outcomes of posterior-only surgical management by titanium mesh versus iliac bone grafts for treating lumbosacral TB. This was a retrospective cohort study. From January 2006 to April 2012, 36 patients with lumbosacral TB were treated at our department. The 36 cases were divided into 2 groups: 17 patients in Group A (titanium mesh) underwent one-stage posterior internal fixation, debridement, and titanium mesh bone fusion. The 19 patients in Group B (iliac bone graft) underwent posterior instrumentation, debridement, and iliac bone graft in a single procedure. The clinical and radiographic results for the 2 groups were analyzed and compared. The mean year of patients was 49.9â±â15.4 months in group A and 55.5â±â12.6 months in group B. All patients were followed up for an average of 47.3â±â8.1 months (range 36-60 months). Spinal TB was completely cured and no intraspinal infection and central nervous system complications of TB infection occurred. Bone fusion was achieved 6.4â±â1.9 months in group A and 7.8â±â2.1 months in group B. There was no significant statistical difference in bone fusion between the 2 groups (Pâ>â.05). The Oswestry Disability Index score (ODI) significantly improved between the preoperative and the last visit in either group. However, no significant difference was observed between the 2 groups at last visit (Pâ>â.05). There were significant differences between groups regarding the postoperative lumbosacral angle and angle correction loss at the final follow-up (Pâ<â.05). The average operative complication rate of Group A was less than that of Group B. Both iliac bone and titanium mesh can effectively construct anterior column defects in posterior surgery. The titanium mesh has the advantage of minor surgical invasion, effective reconstruction of large defects, and ideal sagittal alignment in lumbosacral TB for patients with osteoporosis and poor iliac bone quality.
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Trasplante Óseo/métodos , Ilion/trasplante , Región Lumbosacra/cirugía , Mallas Quirúrgicas , Tuberculosis de la Columna Vertebral/cirugía , Trasplante Óseo/instrumentación , Estudios de Casos y Controles , Femenino , Humanos , Vértebras Lumbares/microbiología , Vértebras Lumbares/cirugía , Región Lumbosacra/microbiología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Sacro/microbiología , Sacro/cirugía , TitanioRESUMEN
OBJECTIVE: To retrospectively analyze the nutritional status and growth and development situation of the children with chronic kidney disease stage 3 to 5 when they were diagnosed at the first visit. METHOD: After searching for the data of all the hospitalized cases during January 2007 to September 2015 in the Department of Nephrology of Children's Hospital Affiliated to the Capital Institute of Pediatrics from the medical record system, data of 37 cases with complete clinical data were collected; all these cases were diagnosed as chronic kidney disease stage 3 to 5 according to the diagnostic criteria.We recorded these children's age, height, weight, body mass index, albumin, blood lipids and acidosis situation when they were first diagnosed, and then, analyzed and summarized their nutritional status and growth and development situation. RESULT: In these 37 cases, 24 cases were boys and 13 cases were girls; 23 cases (62%) were shorter than the third percentile of age-sex-specific height; 18 cases (49%) exhibited lower weight than the third percentile of age-sex-specific weight; 5 cases (13.5%) showed lower BMI than the third percentile of height-age BMI, and 5 cases (13.5%) had obesity. The level of albumin was (37.0±8.7) g/L, and no statistically significant difference was observed within each stage. In all of these cases, 10 cases were hypoalbuminemia (27%), and the difference of its frequency between stage 3-4 and stage 5 was not statistically significant. Triglyceride was (2.2±1.1) mmol/L. The mean level was higher than the normal range, but with no statistically significant difference within each stage; 21 cases (62%) were diagnosed as hypertriglyceridemia, which were more frequent compared with the occurrence of the hypercholesterolemia (32%), the high low density lipoprotein (26%) and the low high density lipoprotein(12%). And the occurrence of decompensated metabolic acidosis in stage 5 (69%) was significantly higher than that in stage 3-4 (38%) (P=0.036 6, <0.05). CONCLUSION: Growth retardation is highly prevalent among the children with chronic kidney disease stage 3 to 5. It would become more frequent and more serious as the stage worsening. Both underweight and obesity could be observed in this kind of children. Low serum albumin level is not a sensitive indicator of malnutrition in these children. The mean value of triglyceride in the children with chronic kidney disease was higher than the normal range. And hypertriglyceridemia is the most common abnormal lipid metabolism in our study.
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Estado Nutricional , Insuficiencia Renal Crónica , Índice de Masa Corporal , Peso Corporal , Niño , Femenino , Humanos , Masculino , Obesidad , Valores de Referencia , Estudios Retrospectivos , Delgadez , TriglicéridosRESUMEN
Euroleon coreanus (Okamoto) is widely distributed in China, and the larval stage can be treated as traditional Chinese medicine. However, the host-bacterium relationship remains unexplored, as there is a lack of knowledge on the microbial community of ant lions. Hence, in the current study, we explored the microbial community of the larval ant lion E. coreanus using Illumina MiSeq sequencing. Results indicated that a total of 10 phyla, 126 genera, and 145 species were characterized from the second instars of E. coreanus, and most of the microbes were classified in the phylum Proteobacteria. Cronobacter muytjensii was the most abundant species characterized in the whole body and gut of E. coreanus, and the unclassified species in the genera Brevundimonas and Lactobacillus were relatively more abundant in the head and carcass. In addition, no Wolbachia-like bacteria were detected, whereas bacteria like Francisella tularensis subsp. Holarctica OSU18 and unclassified Rickettsiella were first identified in ant lion E. coreanus.
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Insectos/microbiología , Animales , Bacterias , China , Larva/microbiologíaRESUMEN
Karyotype analysis in plants helps to reveal the affinity relationships of species and their genetic evolution. The current study aimed to observe chromosome karyotypes and structures of Hyacinthus orientalis. Twenty hyacinth cultivars were introduced from Holland, and their water-cultivated root tips were used as experimental samples. A solution of colchicine (0.02%) and 8-hydroxyquinoline (0.02 M) was used as a 20-h pre-treatment. Subsequently, Carnot I was used for fixation and 45% acetic acid was used for dissociation. The squash method was selected to prepare chromosome spreads for microscopic observation. The basic chromosome number of the hyacinth cultivar was 8, and the number of chromosomes in the diploid, triploid, tetraploid, and aneuploid cultivars was 16, 23, 24, 31, and 32, respectively. The L-type chromosome was predominant in the chromosomal composition. The hyacinth satellite was located on the short arm in numbers equivalent to the ploidy. This satellite is located on the middle-sized chromosome in the fourth group of chromosomes, demonstrating that Hyacinthus has a more primitive evolution than Lilium and Polygonatum. Among 20 hyacinth cultivars, 'Fondant' had the highest level of evolution and a maximum asymmetric coefficient of 61.69%. Moreover, the ratio between the shortest and longest chromosomes in this cultivar was 4.40, and its karyotype was type 2C. This study may elucidate long-term homonym and synonym phenomena. It may also provide a method of cytological identification as well as direct proof of the high outcross compatibility between hyacinth cultivars.
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Hyacinthus/genética , Aberraciones Cromosómicas , Cromosomas de las Plantas , Evolución Molecular , Cariotipo , Raíces de Plantas/genéticaRESUMEN
INTRODUCTION: Stillbirth remains a devastating health issue with 26,000 stillbirths occurring annually in the United States. Formalin-fixed, paraffin-embedded (FFPE) umbilical cord samples are available for many stillbirths. Our aim was to validate the use of these samples in identifying genetic variations in stillbirth through microarray analysis. METHODS: This is a retrospective case-control study from a single institution of stillbirths ≥ 23 weeks gestational age and control liveborn infants. Fetal genomic DNA was extracted from FFPE umbilical cord samples of stillborn and control placentas, and genotyping was performed using the Illumina HumanOmniExpresss-12v1 Beadchip. Array results were verified with qPCR. RESULTS: 31 case-specific CNVs (17 deletions and 14 amplifications) with an average size of 294 kb for amplifications and 74 kb for deletions were identified among 94 FFPE samples (86 cases; 8 controls). In total 38 (44%) of the stillbirth samples had a CNV detected. Validation of a subset of microarray findings with qPCR confirmed deletions on 1p (2 cases), 11q (4 cases) and amplifications on 18 (1 case). Placental underperfusion changes were seen in stillborns with deletions on 1p, a region containing complement regulatory genes which have been shown to play a role in preeclampsia. DISCUSSION: This study validated the use of archived FFPE umbilical cord samples for genome-wide copy number profiling in stillbirths, and demonstrates specific CNV deletions and amplifications. Microarray analysis in an expanded cohort of stillbirth FFPE samples has the potential to identify biomarkers involved in stillbirth pathogenesis.
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Aberraciones Cromosómicas , Variaciones en el Número de Copia de ADN , Placenta/patología , Insuficiencia Placentaria/genética , Mortinato/genética , Cordón Umbilical/patología , Estudios de Casos y Controles , Femenino , Perfilación de la Expresión Génica , Genotipo , Humanos , Masculino , Insuficiencia Placentaria/patología , Embarazo , Estudios RetrospectivosRESUMEN
Dental caries remains one of the most common infectious diseases of humankind, which develops slowly throughout life, affecting children, adolescents, and adults. A vaccine against caries is urgently needed. We previously developed recombinant flagellin as a mucosal adjuvant for anti-Streptococcus mutans vaccines by nasal immunization. Furthermore, we demonstrated a fusion protein strategy that combined flagellin and the target surface adhesion protein (PAc) in a single construct. This construct enhanced specific IgA responses in oral fluids and provided improved prophylactic protection against caries. In the present study, we observed prolonged progression of dental caries in rats after S. mutans Ingbritt challenge. In addition, we observed a therapeutic effect of the flagellin-PAc fusion protein (KF-rPAc) against dental caries as a mucosal vaccine with a new immunization protocol. The present study demonstrated that KF-rPAc by nasal immunization can promote PAc-specific systemic and mucosal antibody responses and inhibit dental caries progression efficiently after the implant of S. mutans into the oral cavity of the rats. The rats immunized with KF-rPAc exhibited 53.9% caries reduction compared with the sham-immunized rats. Our data support the concept of administration of KF-rPAc to humans after infection and even caries that has begun to alleviate caries progression. In conclusion, our study demonstrated that KF-rPAc could be used as an anticaries therapeutic mucosal vaccine.
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Vacunas Bacterianas , Caries Dental/prevención & control , Flagelina/inmunología , Proteínas Recombinantes de Fusión/inmunología , Administración Intranasal , Animales , Anticuerpos Antibacterianos/inmunología , Vacunas Bacterianas/administración & dosificación , Caries Dental/microbiología , Esmalte Dental/patología , Dentina/patología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Femenino , Inmunización , Inmunoglobulina A Secretora/análisis , Inmunoglobulina A Secretora/sangre , Inmunoglobulina G/análisis , Inmunoglobulina G/sangre , Distribución Aleatoria , Ratas , Ratas Wistar , Saliva/inmunología , Organismos Libres de Patógenos Específicos , Streptococcus mutans/inmunologíaRESUMEN
BACKGROUND: Sleep fragmentation during late gestation (LG-SF) is one of the major perturbations associated with sleep apnea and other sleep disorders during pregnancy. We have previously shown that LG-SF induces metabolic dysfunction in offspring mice during adulthood. OBJECTIVES: To investigate the effects of late LG-SF on metabolic homeostasis in offspring and to determine the effects of LG-SF on the epigenome of visceral white adipose tissue (VWAT) in the offspring. METHODS: Time-pregnant mice were exposed to LG-SF or sleep control during LG (LG-SC) conditions during the last 6 days of gestation. At 24 weeks of age, lipid profiles and metabolic parameters were assessed in the offspring. We performed large-scale DNA methylation analyses using methylated DNA immunoprecipitation (MeDIP) coupled with microarrays (MeDIP-chip) in VWAT of 24-week-old LG-SF and LG-SC offspring (n=8 mice per group). Univariate multiple-testing adjusted statistical analyses were applied to identify differentially methylated regions (DMRs) between the groups. DMRs were mapped to their corresponding genes, and tested for potential overlaps with biological pathways and gene networks. RESULTS: We detected significant increases in body weight (31.7 vs 28.8 g; P=0.001), visceral (642.1 vs 497.0 mg; P=0.002) and subcutaneous (293.1 vs 250.1 mg; P=0.001) fat mass, plasma cholesterol (110.6 vs 87.6 mg dl(-1); P=0.001), triglycerides (87.3 vs 84.1 mg dl(-1); P=0.003) and homeostatic model assessment-insulin resistance values (8.1 vs 6.1; P=0.007) in the LG-SF group. MeDIP analyses revealed that 2148 DMRs (LG-SF vs LG-SC; P<0.0001, model-based analysis of tilling-arrays algorithm). A large proportion of the DMR-associated genes have reported functions that are altered in obesity and metabolic syndrome, such as Cartpt, Akt2, Apoe, Insr1 and so on. Overrepresented pathways and gene networks were related to metabolic regulation and inflammatory response. CONCLUSIONS: Our findings show a major role for epigenomic regulation of pathways associated with the metabolic processes and inflammatory responses in VWAT. LG-SF-induced epigenetic alterations may underlie increases in the susceptibility to obesity and metabolic syndrome in the offspring.
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Tejido Adiposo Blanco/metabolismo , Grasa Intraabdominal/metabolismo , Obesidad/metabolismo , Complicaciones del Embarazo/metabolismo , Efectos Tardíos de la Exposición Prenatal/metabolismo , Privación de Sueño/metabolismo , Animales , Animales Recién Nacidos , Modelos Animales de Enfermedad , Epigenómica , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , EmbarazoRESUMEN
Congenital heart disease (CHD) is the most common type of birth defect. It is suspected that polymorphisms in folate metabolism are associated with an increased risk of CHD, but the conclusion remains unclear. Studies have reported that the MTHFR C677T polymorphism was associated with the development of structural congenital heart malformations. The objective of this study was to conduct a meta-analysis of available studies to identify common polymorphisms in the MTHFR gene in children with CHD and their mothers and to test for an association between genotype and disease. In all, 19 eligible studies comprising 4,219 cases and 20,123 controls were included in this meta-analysis. A significant association was found between the MTHFR C677T polymorphism and CHD risk (OR: 1.26; 95 % CI = 1.06-1.51; p = 0.009) with no strong evidence of heterogeneity (I(2) = 39 %) in the fetal analysis. In the maternal analysis, the MTHFR C677T polymorphism was significantly associated with CHD risk (OR = 1.52; 95 % CI = 1.09-2.11; p = 0.01) with significant heterogeneity (I(2) = 63 %).
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Predisposición Genética a la Enfermedad/epidemiología , Predisposición Genética a la Enfermedad/genética , Cardiopatías Congénitas/epidemiología , Cardiopatías Congénitas/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Polimorfismo de Nucleótido Simple/genética , Femenino , Estudios de Asociación Genética , Marcadores Genéticos/genética , Humanos , Incidencia , Masculino , Factores de RiesgoRESUMEN
Sepsis is characterized as an uncontrolled inflammatory response. Spite et al. (Nature 461(7268):1287-1291, 2009) had demonstrated that resolvin D2, which is derived from docosahexaenoic acid (DHA), improves survival in cecal ligation and puncture (CLP)-initiated sepsis and enhances bacterial clearance without immune suppression. Resolvin D1, which is also derived from DHA and homologous with resolvin D2, is an endogenous anti-inflammatory and proresolving lipid molecule. We sought to investigate the effects of resolvin D1 on sepsis and to explore the mechanism of action. Six-to-eight-week-old male C57BL/6 mice were randomly divided into three groups: the sham group underwent the sham operation followed by tail vein injection of vehicle (0.1 % ethanol); the CLP group received vehicle (0.1 % ethanol) after CLP; the resolvin D1 group received resolvin D1 (100 ng) after CLP. Blood, peritoneal lavage fluid, and organs of mice were harvested 24 h after treatment for cytokine analysis, cell counts, bacterial cultures, histopathological studies, and apoptosis quantification. Compared with the vehicle control group, the survival rate and bacterial clearance of mice with sepsis induced by CLP were improved after resolvin D1 treatment, but the numbers of neutrophils in peritoneal lavage fluid, the inflammatory cytokines, the phosphorylation of the nuclear factor-κB (NF-κB) (P65) pathway, and the apoptosis rate of CD3(+) T lymphocytes of the thymus were suppressed. Resolvin D1 treatment improved survival in mice with sepsis induced by CLP, enhanced organism bacterial clearance, suppressed the increase of the numbers of neutrophils in peritoneal lavage fluid, reduced the release of inflammatory cytokines, and decreased the apoptosis rate of CD3(+) T lymphocytes of the thymus. These results suggest that resolvin D1 may attenuate the degree of inflammatory reaction in sepsis caused by CLP, without harming the host defense response.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Inflamación/prevención & control , Sepsis/tratamiento farmacológico , Sepsis/microbiología , Animales , Apoptosis/efectos de los fármacos , Carga Bacteriana/efectos de los fármacos , Citocinas/sangre , Modelos Animales de Enfermedad , Inflamación/metabolismo , Inflamación/microbiología , Inflamación/patología , Estimación de Kaplan-Meier , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Distribución Aleatoria , Sepsis/metabolismo , Sepsis/patologíaRESUMEN
The objective was to cryopreserve porcine primordial follicles by vitrification and to assess the development of these follicles in xenografts. Ovarian tissues containing primordial follicles were collected from neonatal (15-d-old) piglets. They were vitrified in modified tissue culture medium (TCM)-199 containing 15% (v/v) ethylene glycol, 15% (v/v) dimethylsulfoxide, 20% (v/v) fetal calf serum, and 0, 0.25, or 0.5M sucrose. After 1 wk of storage in liquid nitrogen (LN(2)), the tissues were warmed, and the morphology of follicles and oocytes was examined histologically. After vitrification in sucrose-free medium, there were 50+/-2 (mean+/-SEM; n=10) follicles per tissue, in contrast with 108+/-10 (n=10) in fresh tissues. Losses were attributed to puncturing oocytes during the vitrification-warming process, as oocytes were apparently normal after treatment of the sucrose-free vitrification solution without plunging into LN(2). When tissues were vitrified in sucrose-supplemented medium, loss of oocytes decreased (P<0.05). However, the number of abnormal oocytes having nuclear shrinkage was increased (P<0.05) by the addition of 0.5M sucrose; this occurred in a small number of oocytes treated with sucrose-supplemented vitrification solutions without vitrification. After 2 mo of xenografting of vitrified-warmed tissues in SCID (severe combined immune deficiency) mice, primordial follicles developed to the secondary stage (accompanied by oocyte growth), whereas there was development to the antral stage in xenografts of fresh tissues. In conclusion, primordial follicles from neonatal pigs maintained their developmental ability after vitrification and warming, although their developmental rate was slower than that of the fresh control in xenografts.
Asunto(s)
Criopreservación/veterinaria , Folículo Ovárico/crecimiento & desarrollo , Folículo Ovárico/trasplante , Ovario/trasplante , Porcinos , Trasplante Heterólogo , Animales , Animales Recién Nacidos , Femenino , Calor , Riñón , Masculino , Ratones , Ratones SCID , Oocitos/crecimiento & desarrollo , Oocitos/ultraestructuraRESUMEN
We have isolated 462 bp of sequence termed ovarian-specific promoter 1 (OSP-1) that is part of a retrovirus-like element specifically expressed in the rat ovary. We have evaluated the ability of OSP-1 to activate gene expression in normal and neoplastic cell lines derived from the ovaries of rats and women. We have found that there was marked specificity in the ability of OSP-1 to drive reporter gene expression in an ovarian epithelial cell lineage manner. The expression of herpes simplex virus thymidine kinase (HSV-TK) under OSP-1 control was sufficiently ovarian cancer cell line specific to render ganciclovir approximately 50-fold more toxic in the A2780 human ovarian cancer cell line compared with clones of the HCT-116 and HT-29 colon cancer cell lines. Furthermore, ganciclovir had marked antitumor efficacy in vivo in severe combined immunodeficient mice bearing A2780OSP-1-HSV-TK as a s.c. xenograft. We suggest that these data support the use of OSP-1 as a tool to provide specificity to the gene therapy of ovarian cancer and to drive ovarian-specific oncogene expression for the creation of transgenic mouse models of ovarian cancer.