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BACKGROUND: Spontaneous spheroid culture is a novel three-dimensional (3D) culture strategy for the rapid and efficient selection of progenitor cells. The objectives of this study are to investigate the pluripotency and differentiation capability of spontaneous spheroids from alveolar bone-derived mesenchymal stromal cells (AB-MSCs); compare the advantages of spontaneous spheroids to those of mechanical spheroids; and explore the mechanisms of stemness enhancement during spheroid formation from two-dimensional (2D) cultured cells. METHODS: AB-MSCs were isolated from the alveolar bones of C57BL/6 J mice. Spontaneous spheroids formed in low-adherence specific culture plates. The stemness, proliferation, and multi-differentiation capacities of spheroids and monolayer cultures were investigated by reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, alkaline phosphatase (ALP) activity, and oil-red O staining. The pluripotency difference between the spontaneous and mechanical spheroids was analyzed using RT-qPCR. Hypoxia-inducible factor (HIFs) inhibition experiments were performed to explore the mechanisms of stemness maintenance in AB-MSC spheroids. RESULTS: AB-MSCs successfully formed spontaneous spheroids after 24 h. AB-MSC spheroids were positive for MSC markers and pluripotency markers (Oct4, KLF4, Sox2, and cMyc). Spheroids showed higher Ki67 expression and lower Caspase3 expression at 24 h. Under the corresponding conditions, the spheroids were successfully differentiated into osteogenic and adipogenic lineages. AB-MSC spheroids can induce neural-like cells after neurogenic differentiation. Higher expression of osteogenic markers, adipogenic markers, and neurogenic markers (NF-M, NeuN, and GFAP) was found in spheroids than in the monolayer. Spontaneous spheroids exhibited higher stemness than mechanical spheroids did. HIF-1α and HIF-2α were remarkably upregulated in spheroids. After HIF-1/2α-specific inhibition, spheroid formation was significantly reduced. Moreover, the expression of the pluripotency genes was suppressed. CONCLUSIONS: Spontaneous spheroids from AB-MSCs enhance stemness and pluripotency. HIF-1/2α plays an important role in the stemness regulation of spheroids. AB-MSC spheroids exhibit excellent multi-differentiation capability, which may be a potent therapy for craniomaxillofacial tissue regeneration.
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Animals , Mice , Spheroids, Cellular , Mesenchymal Stem Cells , Osteogenesis/genetics , Stem Cells , Cell Differentiation , Cells, Cultured , Cell Culture Techniques/methods , Hypoxia/metabolism , Mice, Inbred C57BLABSTRACT
Objective@#To determine the association between anthropometric parameters and 20 meter shuttle run test (20 m SRT) score among children and adolescents.@*Methods@#The convenient sampling method was conducted to select 3 192 primary and secondary school students in Baoan District, Shenzhen, based on the street school types school from April to May 2019. Height, weight, 20 m SRT score and general demographic indicators were assessed and collected. Individuals were divided into two groups based on the dose response correlation between different anthropometric parameters (the Z score of height, weight and BMI) and 20 m SRT score analyzed with the restricted cubic spline. The association between the Z score of height and 20 m SRT score was further analyzed using the Spearman correlation analysis. Logistic regression analysis was used to analyze separately the relationship different anthropometric parameters and 20 m SRT score.@*Results@#The Z value of weight and 20 m SRT score showed a non linear dose response association ( P <0.01), the significant but weak linear correlation between the Z value of height and 20 m SRT score ( r=0.06, P <0.05). The prevalence rate on the low level of the Z value of 20 m SRT score in 3 192 children and adolescents was 44.7%, and the gender ( χ 2=14.02, P <0.01) and grade difference ( χ 2=93.28, P <0.01) were both statistically significant. There was no significant relationship between the Z value of height and 20 m SRT score grade among total population, different genders and different grades ( P > 0.05). Compared with the reference group on the Z value of weight ≤-0.23, individuals with the Z value of weight >-0.23 had the low level of 20 m SRT score ( OR =0.61, P <0.05). Compared with the reference group on the Z value of BMI ≤ 0.25, individuals with the Z value of weight >0.25 had the low level of 20 m SRT score ( OR =0.45, P <0.05). Stratified for gender and grade, the above significant relationship on the Z value of weight, Z value of BMI and 20 m SRT score were still observed ( P <0.01).@*Conclusions@#The higher height Z value shows on correlations with 20 m SRT score, but the positive association is found between weight and BMI Z value and the 20 m SRT score. The cardiopulmonary fitness improvement may be more effective among children and adolescents when tuking weight and BMI Z scores into consideration.
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La displasia frontometafisaria 2 (DFM2) es una enfermedad rara causada por una mutación en el gen MAP3K7. En este artículo, se informa sobre un paciente de 7 años con DFM2 causada por una variante nueva de corte y empalme en MAP3K7. El paciente presenta las características frecuentes de la DFM2, pero algunas nunca antes informadas. No se dispone de una descripción sistemática de las características de las imágenes tomográficas de la DFM2. Describimos ciertas diferencias en las características de la DFM2, la bibliografía publicada y las manifestaciones imagenológicas generales de la DFM2. Este caso resalta la importancia del valor clínico de la tomografía computada (TC) y la renderización de volúmenes (VR) en el diagnóstico de la DFM2. Las características de la DFM2 pueden observarse claramente en los estudios tomográficos, lo que señala la gran importancia de la TC para el diagnóstico y el tratamiento precoces de los pacientes con DFM2.
Frontometaphyseal dysplasia 2 (FMD2) is a rare disease caused by MAP3K7 gene mutation. We report a 7-year-old sporadic patient with FMD2 due to a de novo splicing variant in MAP3K7. He has the common characteristics of FMD2 but also has some characteristics that have never been reported, which increases the clinical phenotype of FMD2. Moreover, no systematic description of the imaging characteristics of FMD2 in computed tomography (CT) is available. In the present work, we found some different features of FMD2, reviewed previous literature, and summarized the general imaging manifestations of FMD2. This case emphasizes the important clinical value of CT and VR in the diagnosis of FMD2. We can clearly find the characteristics of FMD2 by CT examination, indicating its great significance for the prompt diagnosis and treatment of FMD2 patients.
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Humans , Male , Child , Osteochondrodysplasias/complications , Osteochondrodysplasias/diagnosis , Osteochondrodysplasias/genetics , Pulmonary Arterial Hypertension , Phenotype , ForeheadABSTRACT
Objective@#To analyze the relationship between outdoor activity, screen time and 20 meter shuttle run test (20 m SRT) score among children and adolescents in Baoan District, Shenzhen, so as to provide scientific basis for the management and promotion of physical health of children and adolescents.@*Methods@#Stratified cluster sampling was used to select students from 14 schools in 6 communities in Baoan District of Shenzhen, from April to May 2019. A self designed questionnaire was used to investigate demographic characteristics, outdoor activity duration and screen time of primary and middle school students. A total of 3 192 primary and secondary school students in grade 1-12 were selected from each school. Weight, height and 20 m SRT were measured.@*Results@#About 985(30.9%) out of 3 192 primary and middle school students were estimated excellent on 20 m SRT test, with boys being higher than that of girls (34.7% vs 26.3%) ( χ 2=26.43, P <0.01). Logistic regression analysis showed that compared with the group with outdoor activity <2 h/d, the excellent and good performers on 20 m SRT with outdoor activity time ≥2 h/d was higher( OR=1.38, 95%CI =1.12-1.70). Compared with students with screen time ≥2 h/d, the excellent and good performers on 20 m SRT with screen time <2 h/d was higher( OR=1.42, 95%CI=1.17-1.71). Compared with students with outdoor activity time < 2 h/d and screen time ≥ 2 h/d, students with outdoor activity time ≥2 h/d and video time < 2 h/d had a higher proportion of excellent score on 20 m SRT( OR=1.97, 95%CI =1.46-2.67).@*Conclusion@#Increasing outdoor activity and reducing screen time are helpful to improve the performance score of 20 m SRT among primary and secondary school students.
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Objective@#To explore the association between factors affecting language development and Chinese dyslexia, providing scientific evidence for prevention and intervention of dyslexia.@*Methods@#Twelve elementary schools were selected in Baoan, Shenzhen. The parents and head teachers of 12 868 children in grade 3-5 were surveyed by the Questionnaire for Children s Reading Ability, the Dyslexia Checklist for Chinese Children and the Pupil Rating Scale Revised Screening for Learning Disabilities.@*Results@#The prevalence rate of dyslexia was 2.71%, with 349 children suffering from dyslexia. Gender, parental education and occupations, family income, whether parents work away from home before their child was 3 years old, average time mother spends with her child daily and number of languages spoken in family had statistical significance on dyslexia(all P <0.05). After adjusting for parental education and occupations, and family income, the children who spent more than 1 hour with their mothers per day had a significantly reduced risk of dyslexia (1-2: OR =0.46; 3-4: OR =0.45; 5-6: OR =0.40; >7 h: OR =0.36, P <0.05); the children living in families where two languages were used for communication had a significantly reduced risk of dyslexia( OR=0.74, 95%CI=0.57-0.96, P =0.02). Children with a history of language development disorders had a significantly increased risk of dyslexia( OR=17.30, 95%CI=7.86-38.09, P <0.01).@*Conclusion@#Increase of time mother spend with their child daily and paying more attention to the children with a history of language development disorders can help to prevent the occurrence of dyslexia.
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Abstract@#The prevalence of overweight and obesity among children in China has entered a new phase of pandemic, with substantial regional differences. There are diverse and complex determinants for overweight and obesity in school age children, it is necessary to take targeted measures towards children of different ages and regions, comprehensively consider the effects of physiological, social, household and behavioral factors and simultaneously strengthen connections between schools, medical institutions and families, so as to actively respond to the severe situation of childhood obesity epidemic.
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Abstract Purpose To analyze the plasma lipid spectrum between healthy control and patients with pancreatic cancer and to select differentially expressed tumor markers for early diagnosis. Methods In total, 20 patents were divided into case group and healthy control group according to surgical pathology. Of almost 1206 plasma lipid molecules harvested from 20 patients were measured by HILIC using the normal phase LC/MS. Heat map presented the relative levels of metabolites and lipids in the healthy control group and patients with pancreatic cancer. The PCA model was constructed to find out the difference in lipid metabolites. The principal components were drawn in a score plot and any clustering tendency could be observed. PLS-DA were performed to distinguish the healthy control group and pancreatic cancer according to the identified lipid profiling datasets. The volcano plot was used to visualize all variables with VIP>1 and presented the important variables with P<0.01 and -FC->2. Results The upregulated lipid metabolites in patients with pancreatic cancer contained 9 lipids; however, the downregulated lipid metabolites contained 79 lipids. Conclusion There were lipid metabolomic differences in patients with pancreatic cancer, which could serve as potential tumor markers for pancreatic cancer.
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Humans , Pancreatic Neoplasms/diagnosis , Early Detection of Cancer , Lipidomics , Biomarkers , Biomarkers, Tumor , Metabolomics , LipidsABSTRACT
As one of the most important aquatic fish, Micropterus salmoides suffers lethal and epidemic disease caused by rhabdovirus at the juvenile stage. In this study, a new strain of M. salmoides rhabdovirus (MSRV) was isolated from Yuhang, Zhejiang Province, China, and named MSRV-YH01. The virus infected the grass carp ovary (GCO) cell line and displayed virion particles with atypical bullet shape, 300–500 nm in length and 100–200 nm in diameter under transmission electron microscopy. The complete genome sequence of this isolate was determined to include 11 526 nucleotides and to encode five classical structural proteins. The construction of the phylogenetic tree indicated that this new isolate is clustered into the Vesiculovirus genus and most closely related to the Siniperca chuatsi rhabdovirus. To explore the potential for a vaccine against MSRV, a glycoprotein (1–458 amino acid residues) of MSRV-YH01 was successfully amplified and cloned into the plasmid pFastBac1. The high-purity recombinant bacmid-glycoprotein was obtained from DH10Bac through screening and identification. Based on polymerase chain reaction (PCR), western blot, and immunofluorescence assay, recombinant virus, including the MSRV-YH01 glycoprotein gene, was produced by transfection of SF9 cells using the pFastBac1-gE2, and then repeatedly amplified to express the glycoprotein protein. We anticipate that this recombinant bacmid system could be used to challenge the silkworm and develop a corresponding oral vaccine for fish.
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Objective @#To review the patients who underwent accurate maxillary and mandibular reconstruction with vascularized osteomyocutaneous flap using virtual surgery and evaluate the postoperative results.@*Methods@# From June of 2009 to December of 2014, ninety-four patients were treated in our department. In virtual surgical planning, maxillectomy and mandibulectomy were simulated and donor site bone was superimposed mirror image of normal maxilla and mandible to complete accurate maxillary and mandibular reconstruction. Maxillectomy, mandibulectomy and bone reconstruction were performed using stereomodels and guided templates to replicate the virtual planning. Actual reconstruction result was compared with that of virtual surgery. The patients were asked to assess their satisfaction with facial appearance at 1 year postoperatively. @*Results@#Good bony unions and occlusion were observed in 94 patients. The actual results were basically in accordance with that of virtual surgery. The actual deviation between postoperative CT and preoperative virtual surgical planning was no more than 1 mm matched by computer software. All patients reported their facial appearance were excellent or good at 1 year postoperatively.@*Conclusion@#The ideal contour of neomaxilla and neomandible and good function with dental rehabilitation can be gained after accurate maxillary and mandibular reconstruction with vascularized osteomyocutaneous flap using virtual surgery.
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@#BACKGROUND: Hospital emergency department (ED) use by patients from residential aged care facilities (RACFs) is not always appropriate, and this calls for interventions to avoid some unnecessary uses. This study aims to compare patterns of ED use by RACF patients with and without a Hospital in the Nursing Home (HiNH) program. METHODS: RACF patients presenting to EDs of a hospital with and a hospital without this program during pre- and post-intervention periods were included. Data on patient demographics and ED presentation characteristics were obtained from the Emergency Department Information System database, and were analysed by descriptive and comparative statistics. RESULTS: In both hospitals, most RACF residents presenting to EDs were aged between 75–94 years, female, triaged at scale 3 to 5, and transferred on weekdays and during working hours. Almost half of them were subsequently admitted to hospitals. In accordance with the ICD-10-AM diagnostic coding system, diagnoses that consistently ranked among the top three reasons for visiting the two hospitals before and after intervention included Chapter XIX: injury and poisoning and Chapter X: respiratory diseases. Associated with the intervention, significant decreases in the numbers of presentations per 1000 RACF beds were identified among patients diagnosed with Chapter XI: digestive diseases [rate ratio (95％CI): 0.09 (0.04, 0.22);P<0.0001] and Chapter XXI: factors influencing health status and contact with health services [rate ratio (95％CI): 0.22 (0.07, 0.66);P=0.007]. CONCLUSION: The HiNH program may reduce the incidence of RACF residents visiting EDs for diagnoses of Chapter XI and Chapter XXI.
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A laccase with a molecular mass of 67 kDa and inhibitory activity toward HIV-1 reverse transcriptase (IC50 = 7.5 M) was isolated from fresh fruiting bodies of the Lentinus edodes (Shiitake mushroom). Its characteristics were compared with those of laccases from cultured mushroom mycelia reported earlier. The laccase was unadsorbed on DEAE-cellulose, Affi-gel blue gel and CM-cellulose, but was adsorbed on Con A-Sepharose. About 50-fold purification was achieved with a 19.2% yield of the enzyme. The activity of the enzyme increased steadily from 20°C to 70°C. The activity disappeared after exposure to the boiling temperature for 10 min. Its optimal pH was 4 and very little enzyme activity remained at and above pH 10. The laccase inhibited HIV-1 reverse transcriptase with an IC50 of 7.5 M, but did not demonstrate any antifungal or anti-proliferative activity.
Subject(s)
Amino Acid Sequence , Anti-HIV Agents/chemistry , Anti-HIV Agents/isolation & purification , Anti-HIV Agents/metabolism , Anti-HIV Agents/pharmacology , Fruiting Bodies, Fungal/enzymology , Fruiting Bodies, Fungal/growth & development , HIV Reverse Transcriptase/antagonists & inhibitors , HIV Reverse Transcriptase/metabolism , HIV-1/enzymology , Hydrogen-Ion Concentration , Laccase/chemistry , Laccase/isolation & purification , Laccase/metabolism , Laccase/pharmacology , Molecular Weight , Mycelium/enzymology , Reverse Transcriptase Inhibitors/chemistry , Reverse Transcriptase Inhibitors/isolation & purification , Reverse Transcriptase Inhibitors/metabolism , Reverse Transcriptase Inhibitors/pharmacology , Shiitake Mushrooms/enzymology , Shiitake Mushrooms/growth & development , Substrate Specificity , TemperatureABSTRACT
Objective: To evaluate the outcome and postoperative reaction of dural substitute (Neuropatch) applying in the treatment of Chiari I malformation(CMI) associated with syringomyelia(SM). Methods:Forty patients of CMI associated with SM were operated in our department from Jul. 2002 to Jul. 2004. All patients underwent posterior cranial fossa decompression and duraplasty. They were divided into two groups, 20 patients being repaired with Neuropatch (Neuropatch group), and the others with autologous fascia lata (fascia group). There were 6 males and 14 females in Neuropatch group and 10 males and 10 females in fascia group. The operations were performed under general anesthesia via suboccipital approach and the extent of posterior cranial fossa decompression ranged from 20 cm2 (5 cm×4 cm) to 35 cm2 (5 cm×7 cm). The removal of posterior arch of atlas depended on the extent of tonsillar herniation, and the dura was opened in Y shape. The Neuropatch was cut into triangular shape, and the same sized autologous fascia lata was used in fascia group. The patches were sutured tightly to the dura matter in each group. The incision was closed layer by layer and drainage was used, if necessary. Antibiotics and hormone were routinely used. The duration of operation, postoperative fever were evaluated, the outcome of the operation was evaluated by Tator scale, and the data were analyzed with statistic software SPSS 10.0. Results: There were12 patients (60%) who suffered from postoperative fever in the Neuropatch group, and 9 patients (45%) in the fascia group(χ2=0.902,P=0.342). Seventeen patients in each group were improved postoperatively. The duration of operation, postoperative fever and antibiotics used were compared between the two groups. No significant difference was found, but the duration of postoperative fever and the time of hormone used were different. There were no postoperative infections that occurred after the follow up for 1 to 2 years, except for one patient in fascia group who developed infective granuloma and recovered later by treatment. Conclusion: Neuropatch is a useful dural substitute for the repair of dural defects in the treatment of CMI associated with syringomyelia.