RESUMO
OBJECTIVES: To analyze the imaging manifestations of common fetal oral masses by ultrasound combined with magnetic resonance imaging (MRI) and to discuss their differential diagnoses. METHODS: A retrospective study of 6 fetuses with oral masses was performed at a tertiary referral center. The imaging features of prenatal ultrasonography and MRI in the diagnosis of fetal oral masses were analyzed. RESULTS: Histopathological examination and/or postpartum ultrasound revealed lymphangioma malformation in 2 fetuses, and mucosal retention cyst, mature teratoma, immature teratoma, and cranial meningocele in 1 fetus, respectively. The teratoma had a characteristic sonographic appearance. In our study, the 4 cases of cystic masses did not have an abnormal vessel architecture. Supplemental MRI revealed a mass effect at the level of the hypopharynx, and in 2 cases with polyhydramnios, the mass obstructed the fetuses' upper airway. Thus, ex-utero intrapartum therapy surgery was performed to secure the newborn's airway. CONCLUSIONS: Oral fetal tumors represent rare congenital malformations. This study shows that a prenatal diagnosis of oral masses is feasible by ultrasound examination. MRI can further confirm the results of ultrasonography and clearly show the relationship between the mass and the hypopharynx. Ultrasonography combined with MRI could, to a large extent, facilitate early detection and appropriate treatment and improve outcome.
Assuntos
Feto , Diagnóstico Pré-Natal , Feminino , Humanos , Recém-Nascido , Imageamento por Ressonância Magnética , Gravidez , Estudos Retrospectivos , Ultrassonografia , Ultrassonografia Pré-NatalRESUMO
ABSTRACT: Eyebrow is one of the most important features of the face; the absence of eyebrows will cause a heavy psychological burden to patients. Existing brow reconstruction techniques are not perfect for every case of eyebrow defect. In this study, a patient who suffered from eyebrows loss after radiotherapy for hemangioma underwent eyebrow reconstruction with the relay transfer of a temporal hairline flap. The entire procedure was performed in the outpatient operating room. The temporal hairline flap was transferred to the eyebrow region in a relay and survived well without necrosis. In the 2-month follow-up, the shape and density of the reconstructed eyebrows were satisfactory. The operation showed this technique was innovative, economic, and recommended as a suitable choice for eyebrow reconstruction, especially in those patients with poor hair transplantation recipient area.
Assuntos
Sobrancelhas , Procedimentos de Cirurgia Plástica , Cabelo , Humanos , Procedimentos de Cirurgia Plástica/métodos , Transplante de Pele , Retalhos CirúrgicosRESUMO
ABSTRACT: Fat fixation is a key step in filling tear trough depression with inferior eyelid orbital septum fat. The ideal position for inferior eyelid fat fixation is to cross the tear trough ligament causing tear trough depression and the orbicularis retaining ligament, with the distal end fixed at the farthest end of the dissected lacuna deep down the inferior orbicularis oculi muscle. Traditional suturing is difficult in the deep narrow lacunae, but a buried guide needle can be used to suture and fix the fat in the deepest lacuna. In this study, 264 patients who underwent tear trough filling using a buried guide needle to fix the released inferior eyelid orbital septum fat from 2017 to 2020 were followed up. The preoperative and postoperative imaging findings were compared to evaluate the effectiveness of the operation and postoperative complications. The inferior eyelid bulging, loose skin, and tear trough depression significantly improved than that before the operation. None of the patients had any severe complications, such as inferior eyelid ectropion, lagophthalmos, scar hyperplasia, and diplopia, in the long term (6âmonths) postoperatively. Five patients showed mild eyelid-eyeball separation and recovered in 1âmonth. Four patients had diplopia, and 3 patients had chemosis; all recovered in 7âdays. The tear trough depression was not corrected completely in 2 patients. The operation showed satisfactory results in the improvement of tear trough depression in addition to alleviation of inferior eyelid bulging and loose inferior eyelid skin that is caused by the traditional inferior eyelid pouch removal.
Assuntos
Blefaroplastia , Ectrópio , Lacerações , Tecido Adiposo/transplante , Blefaroplastia/métodos , Depressão , Diplopia/cirurgia , Ectrópio/cirurgia , Pálpebras/cirurgia , Humanos , Lacerações/cirurgiaRESUMO
Introduction: Genetically, complete hydatidiform mole (CHM) is androgenetic diploid, containing two sets of paternal chromosomes. In most cases, recurrent HM (RHM) is CHM but has diploid biparental chromosome constitution. Case report: We report a mother with RHM, both with biparental diploidy. The mother was compound heterozygous for two variants, c.1720dup, p.(C574Lfs*4) and c.2165A > G, p.(D722G) of the NLRP7 gene, as was a brother who fathered 2 normal pregnancies. Conclusion: The genotype study should be obtained for patients of CHM, even in their first pregnancy, followed by genetic screening for maternal-effect variants in those with biparental moles. This strategy will identify patients in their first pregnancy with HM that have a decreased chance for a normal pregnancy, to allow genetic counseling, perhaps utilizing a donor egg.
Assuntos
Doença Trofoblástica Gestacional , Mola Hidatiforme , Neoplasias Uterinas , Proteínas Adaptadoras de Transdução de Sinal/genética , Feminino , Humanos , Mola Hidatiforme/diagnóstico , Mola Hidatiforme/genética , Masculino , Recidiva Local de Neoplasia , Pais , Gravidez , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/genéticaRESUMO
INTRODUCTION: Although ultrasonography has been reported to have similar diagnostic accuracy to magnetic resonance imaging, it is not a standard imaging modality for cervical cancer. We aimed to summarize the ultrasonographic features of rare primary cervical cancer. METHODS: This was a retrospective study of patients with cervical cancer who were diagnosed between June 2014 and October 2019. They were divided into common-type cervical cancer (ie, cervical squamous cell carcinoma) and rare-type cervical cancer groups including adenocarcinoma, adenosquamous carcinoma, and small cell carcinoma. All patients were staged according to the tumor, nodes, and metastases criteria. RESULTS: Of the 64 patients, the diagnosis was suspected on ultrasonography in 61 (95.3%) patients and missed on ultrasonography in three patients. The tumor size was smaller in the rare-type cervical cancer group (p<0.05). Hypoechoic lesions in common-type cervical cancer and isoechoic lesions accounted for 74.4% (32/43) and 61.9% (13/21) of patients in the rare-type cervical cancer group, respectively (p<0.001). Meanwhile, 67.4% (29/43) of tumors in common-type cervical cancer were exophytic, while 66.7% (14/21) in rare-type cervical cancer were endophytic (p=0.01). Color Doppler blood signals, as compared with normal cervical tissue, were found in all patients. There was good consistency between ultrasonographic and pathologic diagnosis of rare-type cervical cancer (weighted kappa=0.87). CONCLUSIONS: Most patients with rare-type cervical cancer present with isoechoic lesions. The coincidence rate between ultrasonographic and pathologic diagnosis of rare-type cervical cancer is 87%.
Assuntos
Adenocarcinoma/patologia , Carcinoma Adenoescamoso/patologia , Carcinoma de Células Escamosas/patologia , Neoplasias do Colo do Útero/patologia , Adenocarcinoma/diagnóstico por imagem , Adulto , Carcinoma Adenoescamoso/diagnóstico por imagem , Carcinoma de Células Escamosas/diagnóstico por imagem , Feminino , Humanos , Pessoa de Meia-Idade , Ultrassonografia Doppler em Cores/métodos , Ultrassonografia Doppler em Cores/normas , Neoplasias do Colo do Útero/diagnóstico por imagemRESUMO
OBJECTIVE: The incidence of epicanthus is extremely high in Asians. Although many techniques have been introduced to remove the epicanthus, there are still some complications. Herein, we introduced a method involving an incision on the lower palpebral margin combined with a tiny triangular flap for correction of epicanthal folds, which is always performed in combination with double eyelid plasty. METHODS: After making an incision, the ectopic medial canthal ligament was removed. The ligament pulling the medial canthus was cut along the lower palpebral margin incision, so the medial canthus could be repositioned upward. The tiny triangular flap of the medial canthus would naturally move downward and cover the lower palpebral margin incision. The skin folds on the lower palpebral margin were trimmed and sutured. After epicanthoplasty was completed, double eyelid plasty was performed using the incision or non-incision method. RESULTS: A total of 547 epicanthoplasties were performed during the past 7 years. The follow-up time ranged from 1 month to 5 years. Ninety-eight percent of the patients were satisfied with the results of the epicanthoplasty. The scars in the medial canthal area were tiny, the enlargement of the medial canthus was obvious, and the double eyelid line in the medial canthal area was natural and smooth. CONCLUSION: The introduced method is simple in design and can preserve the original shape of the medial canthus while widening the palpebral fissure, which is a particularly suitable approach for epicanthoplasty in Asian patients. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
Assuntos
Blefaroplastia , Aparelho Lacrimal , Povo Asiático , Pálpebras/cirurgia , Humanos , Aparelho Lacrimal/cirurgia , Estudos Retrospectivos , Retalhos Cirúrgicos , Resultado do TratamentoRESUMO
BACKGROUND: "European-style double eyelid" blepharoplasty often leads to a complex deformity, which includes one or more of the following features: a high fold, upper eyelid depression, multiple folds, an obvious scar, blepharoptosis, and lower flap tumidness. However, there is still a lack of comprehensive research on evaluation and treatment strategies for this complex deformity. METHODS: During a 6-year period, 65 patients (106 eyes) with a high fold and upper eyelid depression underwent corrective blepharoplasty. The authors classified this complex deformity as mild, moderate, or severe based on the extent of adhesion and tissue insufficiency, and we performed preaponeurotic fat flap transfer in 47 eyes (44.3%), free fat graft in 35 eyes (33%), and free dermis-fat graft in 24 eyes (22.6%), respectively. RESULTS: The outcomes in 87 eyelids were judged as excellent; and the grading was excellent in 41 eyes (87.2%) that underwent preaponeurotic fat flap transfer, 30 eyes (85.7%) that received a free fat graft, and 16 eyes (66.7%) that received a free dermis-fat graft. The outcomes in only 3 eyes (2.8%) were evaluated as unsatisfactory. CONCLUSIONS: The fundamental reasons behind this deformity were adhesion and tissue insufficiency above the supratarsal crease. The purpose of correction was to reconstruct the gliding system and restore the volume. The authors performed preaponeurotic fat flap transfer, free fat graft, and free dermis-fat graft and achieved satisfactory results. Great improvement in ptosis was achieved by releasing the adhesion and lowering the fold in patients with normal levator muscle function. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
Assuntos
Blefaroplastia/efeitos adversos , Blefaroplastia/métodos , Pálpebras/anormalidades , Pálpebras/cirurgia , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/cirurgia , Adolescente , Adulto , Povo Asiático , Feminino , Humanos , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Adulto JovemRESUMO
Introduction: Congenital diaphragmatic eventration (CDE) is defined as the abnormal elevation of the diaphragm, due to incomplete muscularization of the diaphragm with a thin membranous sheet replacing normal diaphragmatic muscle. Case report: We report a prenatal case with a diaphragmatic mesothelial cyst combined with CDE. Conclusion: A large cystic mass between the thoracic wall and the liver in early pregnancy is highly suggestive of cystic diaphragm.
Assuntos
Diafragma/anormalidades , Diafragma/embriologia , Eventração Diafragmática/diagnóstico , Adulto , Anormalidades Congênitas , Diagnóstico Diferencial , Epitélio/patologia , Feminino , Feto , Humanos , Fígado/embriologia , Masculino , Gravidez , Diagnóstico Pré-Natal , Parede Torácica/embriologia , UltrassonografiaRESUMO
BACKGROUND: In buried suture methods, the levator aponeurosis is fixed to the subcutaneous tissue in the pretarsal region using a suture. However, loosening of the suture occurs frequently and causes regression or disappearance of the double-eyelid folds. To avoid potential loosening of the suture after surgery, we modified the horizontal suture technique commonly used in buried suture double-eyelid blepharoplasty. METHODS: In our procedure, the levator aponeurosis was sutured horizontally, and then the subcutaneous tissue in the pretarsal region was sutured vertically by the same suture. After the two ends of the suture were tied, three tissue layers, namely, the levator aponeurosis, pretarsal fascia, and orbicularis oculi muscle, were fixed together in the pretarsal region. RESULTS: A total of 873 Asian patients underwent double-eyelid blepharoplasty during the past 8 years. No loss of the double-eyelid folds occurred in 563 patients who were followed up for more than six months, and 531 patients, accounting for 94% of the sample, were satisfied with the postoperative results. CONCLUSION: Since the suture was perpendicular to both the levator aponeurosis and the pretarsal orbicularis oculi muscle, pulling on the suture fixation site during blinking was effectively reduced. As a result, regression or disappearance of the double-eyelid folds due to loosening of the suture along the orientation of the muscle fibers was avoided, and the long-term stability of the double-eyelid folds was ensured.
Assuntos
Blefaroplastia , Tornozelo/cirurgia , Blefaroplastia/métodos , Pálpebras/cirurgia , Humanos , Técnicas de Sutura , SuturasRESUMO
Background: Autologous fat grafting is a procedure that treats soft tissue defects by reallocating fat to improve a patient's physical appearance. Imaging methods may be used to evaluate and monitor the grafted fat after transplantation. The goal of imaging is to examine the signal and volume of the grafted fat after autologous fat grafting during the adipose tissue recovery. However, researchers have yet to examine the feasibility of using fat-only imaging to assess the autologous fat graft. Methods: In this prospective and observational study, 46 injected sides in 23 female patients (age 35±7.8 years) were included in the image evaluation. The patients underwent autologous fat grafting surgery with filtered and washed fat. A total of 16, 18, and 12 sides were scanned 7 days, 3 months, and 1 year after fat grafting, respectively. Fat-only images were obtained using Dixon imaging, and then the image quality and contrast of the T1W and T2W were rated to evaluate the application of this method when imaging the autologous fat. The signal and volume of the autologous fat graft were recorded to assess the retention during recovery of the autologous fat tissue. Results: Fat-only T1W magnetic resonance imaging (MRI) was used to identify and delineate grafted fat because this method had better image quality and image differentiation than did T2W MRI. The average signal contrast and retention rate measured 7 days postoperation (28.8%±4.7%; 94.1%±5.8%) was the highest and then decreased at 3 months (16.3%±2.1%; 48.7%±17.3%) and 1 year (3.3%±1.3%, 33.1%±12.9%) after surgery. There were statistically significant differences between the signal and volume retention measurements at each postoperative recovery phase. Conclusions: The T1W fat-only images produced by Dixon MRI is a feasible approach for identifying grafted fat and measure postoperative changes during clinical evaluation. We found a significant decrease in signal contrast and volume of the grafted fat from the surgery date to 3 months postoperation and from 3 months to 1-year postoperation.
RESUMO
Early-stage ovarian serous carcinoma is usually difficult to detect in clinical practice. The profiling of protein expression in high-grade serous carcinoma (HGSC) and low-grade serous carcinoma (LGSC) would provide important information for diagnoses and chemotherapy. Here, we performed proteomic profiling of specimens from 13 HGSC and 7 LGSC patients by iTRAQ. A total of 323 proteins that were differentially expressed were identified. After immunohistochemical confirmation of expressed proteins in 166 clinical tissues, asparagine synthetase (ASNS) and filamin A (FLNA) were selected for further functional study. Cisplatin-sensitive (CS; ASNShigh and FLNAlow) and cisplatin-resistant (CR; ASNSlow and FLNAhigh) SKOV3 and OVCAR3 ovarian cancer cell lines were used for subsequent in vitro and in vivo experiments. Notably, ASNS overexpression (ASNS+) or FLNA knockdown (shFLNA) enabled cisplatin-induced apoptosis and autophagy in CR cells. However, ASNS+ and shFLNA promoted and attenuated tumor growth, respectively. In CS cells, ASNS knockdown (shASNS) attenuated clonogenicity, cell proliferation, and the epithelial-mesenchymal transition, whereas FLNA overexpression (FLNA+) protected cells from cisplatin. In vivo, cisplatin resistance was attenuated in mice xenografted with ASNS+, shFLNA, or ASNS+-shFLNA CR cells, whereas xenografts of shASNS or FLNA+ CS cells exhibited resistance to cisplatin. Clinically, all HGSC patients (83/83) responded to cisplatin, while 6 in 41 LGSC patients exhibited cisplatin resistance. These findings identify ASNS and FLNA as distinct biomarkers for HGSC and LGSC, which may have potential value in the prognosis and clinical treatment of serous carcinoma.
RESUMO
Bone repair using tissue-engineered bone (TEB) in a large defect or accompanied by a poor recipient vascular bed is a long-standing challenge. Surgical vascular carrier patterns of vascular bundle (VB) and arteriovenous loop (AV loop) have been shown to improve the vascularization and repair capacity of TEB. However, the effects of these different vascular carrier patterns on angiogenesis and osteogenesis in TEB have never been evaluated. Here, TEB was constructed with bone marrow mesenchymal stem cells (BMSCs) and ß-TCP and prevascularized by the VB or AV loop technique in beagle dogs. The vascularization and bone formation in TEB were quantitatively compared using Microfil perfusion, histological examination and CT and micro-CT analyses. The distribution and constitution of the neovasculature were analysed to determine the underlying mechanism of angiogenesis. The results showed that prevascularized TEB generated bone tissue faster and more homogeneously than untreated TEB. The VB technique was found to strike a better balance between bone regeneration and ß-TCP scaffold degradation than the AV loop strategy, which resulted in more vascularization but less bone yield, due to faster degradation of the ß-TCP scaffold. This study indicates that a suitable triangular relationship, composed of bone regeneration, scaffold degradation and vasculature, is critical to TEB construction. Copyright © 2015 John Wiley & Sons, Ltd.
Assuntos
Regeneração Óssea , Osso e Ossos/fisiologia , Células-Tronco Mesenquimais/citologia , Osteogênese , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Células da Medula Óssea/citologia , Fosfatos de Cálcio/química , Células Cultivadas , Cães , Masculino , Neovascularização Fisiológica , Tomografia Computadorizada por Raios X , Cicatrização , Microtomografia por Raio-XRESUMO
Survivin is the smallest member of mammalian IAP (inhibitor of apoptosis) family. It is ubiquitous during embryonic development but is not expressed in normal post-natal tissues, except the thymus, colonic epithelial cells and CD34+ hematopoietic stem cells. However, its expression is upregulated during neoplastic transformation in both solid organ and hematological malignancies, including leukemia and lymphoma. In this study, we used RNA interference with short hairpin RNA (shRNA) technique to inhibit survivin expression in a Burkitt's lymphoma cell line Raji and validated its effects on apoptosis and cell proliferation. A survivin-shRNA expression vector were constructed and introduced into Raji cells. Expression of survivin mRNA and protein was assessed by RT-PCR and western blot analysis. Apoptosis index of transfected cells was quantified by flow cytometry and cell proliferation was enumerated by trypan blue exclusion. In Raji cells treated with survivin-shRNA expression vector, survivin mRNA levels were significantly reduced by 67.14% (transient transfection) and 64.28% (stable transfection) respectively, compared with control-shRNA treated group and PBS treated group (p<0.05). The levels of survivin protein were significantly reduced by 62.50% (transient transfection) and 60.93% (stable transfection), compared with the two control groups (p<0.05). Apoptosis index was significantly increased during transient transfection and stable transfection, respectively 31.20+/-2.45% and 29.40+/-1.72% (p<0.05). Survivin-shRNA inhibited the proliferation of Raji cells of stable transfection. In conclusion, the vector-based survivin-shRNA can effectively reduce the expression of survivin gene and induce apoptosis and growth inhibition of transfected Raji cells. We suggest that survivin can be regarded as an ideal target for new anticancer intervention of NHL.
Assuntos
Apoptose/efeitos dos fármacos , Linfoma de Burkitt/terapia , Proteínas Associadas aos Microtúbulos/antagonistas & inibidores , Proteínas de Neoplasias/antagonistas & inibidores , RNA Interferente Pequeno/farmacologia , Linfoma de Burkitt/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Humanos , Proteínas Inibidoras de Apoptose , Proteínas Associadas aos Microtúbulos/genética , Proteínas de Neoplasias/genética , RNA Mensageiro/análise , SurvivinaRESUMO
Allogeneic bone marrow mesenchymal stem cell (allo-BMSC)-based tissue-engineered bone (TEB) has great potential for bone defect repair. However, the immunogenicities and biological roles of allo-BMSCs are still controversial. In this study, we established an animal model of critical-sized mandibular defect in beagle dogs and compared the repairing effects of allo-BMSC-based TEB with autogenic BMSC (auto-BMSC)-based TEB without the administration of immunosuppressants. During the first 2 weeks postimplantation, a transient immune response in the allo-BMSC group was detected with an increase in proinflammation cytokines TNF-α, IFN-γ, and IL-2, a declination of anti-inflammation cytokine IL-10, and an increase in percentages of CD4(+) and CD8(+) T-cell subsets in peripheral blood. Nevertheless, there was no significant difference in bone union achievement, bone mineral density, and biomechanical properties between the two groups at 12 and 24 weeks postimplantation. Further subcutaneous implantation of allo-BMSCs/scaffold also exhibited the similar transient immune responses in the first 2 weeks postimplantation but followed by a decreased bone formation at 4 and 8 weeks postimplantation. These findings indicate that allo-BMSCs can induce a transient immunoreaction, which may temporally delay the osteogenesis of allo-BMSC/scaffold complex in early stage of in vivo implantation, whereas the long-term engineered bone formation was not affected.
Assuntos
Células da Medula Óssea/citologia , Regeneração Óssea/imunologia , Células-Tronco Mesenquimais/imunologia , Osteogênese/fisiologia , Engenharia Tecidual , Animais , Células da Medula Óssea/imunologia , Células Cultivadas , Cães , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Subpopulações de Linfócitos T/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Transplante Homólogo , Cicatrização/fisiologiaRESUMO
OBJECTIVE: To explore the clinical characteristics of Wolman disease and diagnostic methods using enzymatic and molecular analysis. METHOD: Lysosomal acid lipase activity was measured using 4-methylumbelliferyl oleate in the leukocytes of an infant suspected of Wolman disease and LIPA gene mutational analysis was performed by PCR and direct sequencing in the proband and his parents. After the diagnosis was confirmed, the clinical, biochemical, radiological and histopathological findings in this case of Wolman disease were retrospectively reviewed. RESULT: The sixteen-day-old boy was failing to thrive with progressive vomiting, abdominal distention and hepatosplenomegaly. Abdominal X-ray revealed adrenal calcifications which were confirmed on abdominal CT scan. Xanthomatosis were observed on enlarged liver, spleen and lymph nodes during abdominal surgery. Liver and lymph node biopsy showed foamy histiocytes. The lysosomal acid lipase activity in leukocytes was 3.5 nmol/(mg·h) [control 35.5 - 105.8 nmol/(mg·h)]. Serum chitotriosidase activity was 315.8 nmol/(ml·h) [control 0 - 53 nmol/(ml·h)]. The patient was homozygote for a novel insert mutation allele c.318 ins T, p. Phe106fsX4 in exon 4 on LIPA gene. His both parents were carriers of the mutation. CONCLUSION: The clinical features of Wolman disease include early onset of vomiting, abdominal distention, growth failure, hepatosplenomegaly and bilateral adrenal calcification after birth. A plain abdominal X-ray film should be taken to check for the typical pattern of adrenal calcification in suspected cases of Wolman disease. The enzymatic and molecular analyses of lysosomal acid lipase can confirm the diagnosis of Wolman disease.
Assuntos
Leucócitos/enzimologia , Lipase/sangue , Mutação , Esterol Esterase/genética , Doença de Wolman/diagnóstico , Doença de Wolman/genética , Doenças das Glândulas Suprarrenais/etiologia , Doenças das Glândulas Suprarrenais/patologia , Éxons , Humanos , Recém-Nascido , Lipase/genética , Fígado/patologia , Lisossomos/enzimologia , Lisossomos/genética , Masculino , Reação em Cadeia da Polimerase , Esplenomegalia/patologia , Tomografia Computadorizada por Raios X , Doença de Wolman/enzimologia , Doença de Wolman/patologiaRESUMO
Bone tissue engineering (BTE) has been demonstrated an effective approach to generate bone tissue and repair bone defect in ectopic and orthotopic sites. The strategy of using a prevascularized tissue-engineered bone grafts (TEBG) fabricated ectopically to repair bone defects, which is called live bone graft surgery, has not been reported. And the quantitative advantages of vascularization and osteogenic environment in promoting engineered bone formation have not been defined yet. In the current study we generated a tissue engineered bone flap with a vascular pedicle of saphenous arteriovenous in which an organized vascular network was observed after 4 weeks implantation, and followed by a successful repaire of fibular defect in beagle dogs. Besides, after a 9 months long term observation of engineered bone formation in ectopic and orthotopic sites, four CHA (coral hydroxyapatite) scaffold groups were evaluated by CT (computed tomography) analysis. By the comparison of bone formation and scaffold degradation between different groups, the influences of vascularization and micro-environment on tissue engineered bone were quantitatively analyzed. The results showed that in the first 3 months vascularization improved engineered bone formation by 2 times of non-vascular group and bone defect micro-environment improved it by 3 times of ectopic group, and the CHA-scaffold degradation was accelerated as well.
Assuntos
Antozoários , Osso e Ossos/citologia , Hidroxiapatitas , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual , Angiografia , Animais , Osso e Ossos/irrigação sanguínea , Diferenciação Celular , Cães , Tomografia Computadorizada por Raios XRESUMO
OBJECTIVE: Dermal papillae cells are widely applied to reconstruction of tissue engineered hair follicle and skin. To investigate the difference of the biological characteristics of dermal papillae cells cultured with keratinocyte medium (KM) and normal medium (NM), and to determine whether it is feasible for the reconstruction of tissue engineered hair follicle using dermal papillae cells cultured in KM. METHODS: Scalp samples were obtained in rhytidectomy procedure. Dermal papillae were isolated by two steps digestive treatment, then cultured with KM and NM in two groups. The time of dermal papillae adherence and cell outgrowth was recorded and the rate of dermal papillae adherence was determined after 5 days. As well as, the difference of cell morphology was observed through inverted phase contrast microscope. The maximum generations were determined in two groups and the cell sheets were observed by HE staining. In third-generation cells, the number of aggregates in every dish and the proliferation by MTT were compared between two groups. Meanwhile, the expression of a-smooth muscle actin (alpha-SMA) and ALP were detected by immunofluorescence and specific staining in two groups. RESULTS: Dermal papillae of KM group had a higher rate of adherence and fast outgrowth. The rates of adherence were 54.17% and 36.78% in KM group and in NM group, respectively. In KM group, cells adhered after 24 hours and outgrew after 64 hours. While, cells adhered after 48 hours and outgrew after 80 hours in NM group. The cells were bigger in NM group than in KM group. In third-generation cells, 3.06 +/- 1.12 and 9.25 +/- 1.73 aggregates formed in NM group and KM group, respectively, the difference was significant (P < 0.05). In addition, cells could form cell sheets which were muti-layers in KM group. Mostly 7 and 15 generations could been subcultured in NM group and KM group, respectively. The result of MTT indicated that cells proliferated more actively in KM group; absorbance value of KM group was significantly higher than that of NM group after 7 days (P < 0.05). The positive of alpha-SMA were detected in the third-generation cells of both groups. Occasionally a little few cells expressed ALP with (987 +/- 146) microm2 positive area in the sixth-generation cells of NM group. However, the cells still expressed ALP with (8 757 +/- 558) microm2 positive area in the fourteenth-generation cells of KM group and the difference was significant (P < 0.05). CONCLUSION: Cells proliferate actively and aggregate obviously and could been subcultured more generations in KM. Therefore, culturing dermal papillae cells with KM is feasible for the reconstruction of tissue engineered hair follicle.
Assuntos
Derme/citologia , Queratinócitos/citologia , Pele/citologia , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Separação Celular , Células Cultivadas , Humanos , Engenharia Tecidual/métodosRESUMO
OBJECTIVE: To transfect a short hairpin RNA (shRNA) against survivin gene into human T lymphoblastic leukemia cell line Jurkat, and to explore the effects on apoptosis and proliferation of transfected cells. METHODS: The survivin-shRNA expression vector were constructed and transfected into Jurkat cells. Expression of survivin mRNA and protein were assessed by RT-PCR and Western blot analysis respectively. Apoptosis index of transfected Jurkat cells was quantified by flow cytometry. The potential of cell proliferation was described by cell growth curves. RESULTS: In survivin-shRNA transfected Jurkat cells, survivin mRNA levels were significantly reduced by 66.67% ( transient transfection) and 60.69% ( stable transfection) respectively, compared with that in control-shRNA treated group and PBS treated group (P < 0.05); and the levels of survivin protein were significantly reduced by 63.41% (transient transfection) and 60.18% (stable transfection), compared with that in the two control groups (P < 0.05). Apoptosis index was significantly increased during both transient and stable transfection, respectively [(22. 41 +/- 2.83)% and (20.73 +/- 2.56)% (P < 0.05)]. Survivin-shRNA also inhibited the proliferation of Jurkat cells. CONCLUSIONS: Vector-based survivin-shRNA can effectively reduce the expression of survivin gene, induce apoptosis