RESUMO
Objective: To establish a three-dimensional model of middle ear-eustachian tube based on Chinese digital visual human dataset, and the deformation and pressure changes of the middle ear-eustachian tube system after eustachian tube opening are simulated by computer numerical simulation. Methods: The first female Chinese Digital Visual Human data was adopted. The images were imported by Amira image processing software, and the images were segmented by Geomagic software to form a three-dimensional model of middle ear-eustachian tube system, including eustachian tube, tympanum, tympanic membrane, auditory ossicles, and mastoid air cells system. The 3D model was imported into Hypermesh software for meshing and analysis. The structural mechanics calculation was carried out by Abaqus, and gas flow was simulated by Xflow. The tissue deformation and middle ear pressure changes during eustachian tube opening were numerically simulated by fluid-solid coupling algorithm. Several pressure monitoring points including tympanum, mastoid, tympanic isthmus, and external auditory canal were set up in the model, and the pressure changes of each monitoring point were recorded and compared. Results: In this study, a three-dimensional model of middle ear-eustachian tube and a numerical simulation model of middle ear ventilation were established, including eustachian tube, tympanum, mastoid air cells, tympanic membrane, and auditory ossicles. The dynamic changes of the model after ventilation could be divided into five stages according to the pressure. In addition, the pressure changes of tympanum and tympanic isthmus were basically synchronous, and the pressure changes of mastoid air cells system were later than that of tympanum and tympanic isthmus, which verified the pressure buffering effect of mastoid. The extracted pressure curve of the external auditory canal was basically consistent with that of tympanometry in terms of value and trend, which verified the effectiveness of the model. Conclusions: The numerical simulation model of middle ear-eustachian tube ventilation established in this paper can simulate the tissue deformation and middle ear pressure changes after eustachian tube opening, and its accuracy and effectiveness are also verified. This not only lays a foundation for further research, but also provides a new research method for the study of middle ear ventilation.
Assuntos
Feminino , Humanos , China , Orelha Média , Tuba Auditiva , Corpo Humano , Ventilação da Orelha MédiaRESUMO
A boy aged 2 months (born at 36 weeks of gestation) was admitted due to cough and dyspnea. After admission, he was found to have persistent hypertension, proteinuria, and persistent convulsion, and imaging examination showed extensive calcification of the aorta and major branches and stenosis of local lumens of the abdominal aorta and the right renal artery with increased blood flow velocity. The boy was admitted during the neonatal period due to wet lung and pulmonary arterial hypertension and was found to have hypertension and proteinuria. High-throughput whole-exome sequencing was performed and found two compound heterozygous mutations in the ENPP1 gene from his parents, c.130C>T (p.Q44X) and c.1112A>T (p.Y371F). c.130C>T was a nonsense mutation, which could cause partial deletion of protein from 44 amino acids, and was defined as a primary pathogenic mutation. c.1112A>T was a missense mutation which had been reported as a pathogenic mutation associated with idiopathic infantile arterial calcification (IIAC). Therefore, he was diagnosed with IIAC. He was given phosphonate drugs, antihypertensive drugs, anticonvulsion treatment, and respiratory support. Blood pressure was maintained at the upper limit of normal value. There was no deterioration of arterial calcification. It is concluded that IIAC should be considered for infants with persistent hypertension and extensive vascular calcification, and imaging and genetic examinations should be performed as early as possible to make a confirmed diagnosis.
Assuntos
Humanos , Lactente , Masculino , Hipertensão , Recém-Nascido Prematuro , Mutação , Calcificação VascularRESUMO
<p><b>BACKGROUND</b>L-proline is a natural, nontoxic cryoprotectant that helps cells and tissues to tolerate freezing in a variety of plants and animals. The use of L-proline in mammalian oocyte cryopreservation is rare. In this study, we explored the cryobiological characteristics of L-proline and evaluated its protective effect in mouse oocyte cryopreservation.</p><p><b>METHODS</b>The freezing property of L-proline was detected by Raman spectroscopy and osmometer. Mature oocytes obtained from 8-week-old B6D2F1 mice were vitrified in a solution consisting various concentration of L-proline with a reduced proportion of dimethyl sulfoxide (DMSO) and ethylene glycol (EG), comparing with the control group (15% DMSO and 15% EG without L-proline). The survival rate, 5-methylcytosine (5-mC) expression, fertilization rate, two-cell rate, and blastocyst rate in vitro were assessed by immunofluorescence and in vitro fertilization. Data were analyzed by Chi-square test.</p><p><b>RESULTS</b>L-proline can penetrate the oocyte membrane within 1 min. The osmotic pressure of 2.00 mol/L L-proline mixture is similar to that of the control group. The survival rate of the postthawed oocyte in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG is significantly higher than that of the control group. There is no difference of 5-mC expression between the L-proline combination groups and control. The fertilization rate, two-cell rate, and blastocyst rate in vitro from oocyte vitrified in 2.00 mol/L L-proline combining 7.5% DMSO and 10% EG solution are similar to that of control.</p><p><b>CONCLUSIONS</b>It indicated that an appropriate concentration of L-proline can improve the cryopreservation efficiency of mouse oocytes with low concentrations of DMSO and EG, which may be applicable to human oocyte vitrification.</p>
Assuntos
Animais , Feminino , Masculino , Camundongos , Criopreservação , Métodos , Crioprotetores , Farmacologia , Fertilização in vitro , Concentração de Íons de Hidrogênio , Oócitos , Pressão Osmótica , Prolina , Farmacologia , Análise Espectral Raman , VitrificaçãoRESUMO
<p><b>OBJECTIVE</b>To identify the single nucleotide polymorphisms (SNPs) of the alpha2-Heremans-Schmid glycoprotein (AHSG) gene and assess their association with the AHSG serum level.</p><p><b>METHODS</b>The SNPs of the AHSG gene were identified from 30 unrelated Han individuals from Guangzhou area by resequencing. Linkage disequilibrium(LD) was performed to observe the linkage disequilibrium pattern. Then tagSNPs were genotyped in 192 Han individuals from Beijing and 424 Han individuals from Guangzhou area. Finally, luciferase reporter gene assay was performed to determine whether the SNPs affected the promoter activity. Serum AHSG concentrations were measured in the 192 subjects from Beijing using ELISA.</p><p><b>RESULTS</b>Eight SNPs were detected in total. The linkage disequilibrium profile in the Guangzhou Han population was different from that in the Beijing Han population. However, the allele and genotype frequencies of tagSNPs between the two Han populations were not significantly different. The reporter gene assay showed that the -799A allele had significantly higher promoter activity than the -799T allele. Multiple regression analysis revealed that only the rs2248690 SNP was an independent contributor to serum AHAG concentration.</p><p><b>CONCLUSION</b>The rs2248690 SNP in the promoter region of the AHSG gene might affect the AHSG gene transcription.</p>
Assuntos
Proteínas Sanguíneas , Genética , Metabolismo , Ensaio de Imunoadsorção Enzimática , Genótipo , Desequilíbrio de Ligação , Polimorfismo de Nucleotídeo Único , Genética , Soro , Química , alfa-2-Glicoproteína-HSRESUMO
<p><b>OBJECTIVE</b>To determine the expression of immunoglobulins in HT-29 cells (an established colon cancer cell line, and explore their effect on the biological activities of the cancer cells.)</p><p><b>METHODS</b>The transcripts of variable regions of immunoglobulin heavy chains in HT-29 cells were detected by RT-PCR. Antisense CDR3 (specific to HT-29)-pIRES 1 neo vector was constructed, then transfected into HT-29 cells by electroporation. Programmed cell death and growth inhibition of HT-29 cells were detected by FCM and MTT, respectively.</p><p><b>RESULTS</b>The transcripts of Ig heavy chain (V(H) CDR3 region) were expressed in HT-29 cells. Moreover, they showed a monoclonal characteristic after being sequenced. After transfection of the antisense vector of CDR3 (specific to HT-29)-pIRES 1 neo, expression level of Ig in HT-29 cells was significantly decreased, and growth inhibition (P < 0.05) and apoptosis (P < 0.01) were induced.</p><p><b>CONCLUSION</b>These results suggest that tumor derived Ig could promote the survival and growth of tumor cells.</p>
Assuntos
Humanos , Apoptose , Proliferação de Células , Regiões Determinantes de Complementaridade , Genética , DNA Antissenso , Genética , Eletroporação , Vetores Genéticos , Células HT29 , Células HeLa , Imunoglobulina G , Metabolismo , Cadeias Pesadas de Imunoglobulinas , Genética , Imunoglobulina M , Metabolismo , Região Variável de Imunoglobulina , Genética , Imunoglobulinas , Metabolismo , RNA Mensageiro , Genética , Proteínas Recombinantes , Genética , TransfecçãoRESUMO
<p><b>OBJECTIVE</b>To study the histochemical staining in the diagnosis of osteosarcoma.</p><p><b>METHODS</b>To compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification.</p><p><b>RESULTS</b>With modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining.</p><p><b>CONCLUSION</b>The modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.</p>
Assuntos
Humanos , Neoplasias Ósseas , Patologia , Histocitoquímica , Osteossarcoma , Patologia , Coloração e Rotulagem , MétodosRESUMO
<p><b>OBJECTIVE</b>To prepare the polyclonal anti-peptide antibody against chemokine-like factor1 (CKLF1) and apply it to the expression and functional studies of CKLF1.</p><p><b>METHODS</b>CKLF1 was analyzed with bioinformatics methods. The 16 amino acids sequence peptide was selected from CKLF1 C terminal end. Antibody was raised by immunizing rabbits with the peptide conjugated to keyhole limpet hemocyanin (KLH).</p><p><b>RESULTS</b>A high titer polycolonal antibody was obtained from the rabbit against the peptide. ELISA analysis proved that the titer of rabbit serum against anti-peptide of CKLF1 was up to 10(-4). Western blot analysis revealed that it could react not only with recombinant CKLF1 expressed in a cell-Free Protein Biosynthesis System and Drosophila S2 cells, but also recognize the endogenous CKLFs in the tissue array. Positive staining was detected in the normal bronchial cartilage, gastric mucosa, and gastric smooth muscle tissues. Normal rectum and well-differentiated rectal carcinoma showed strong positive staining, but the poor-differentiated rectal carcinoma samples revealed negative staining.</p><p><b>CONCLUSION</b>The anti-peptide antibody can specifically recognize CKLFs and may be a useful reagent for the detection of CKLF1.</p>