RESUMEN
In the present study we have successfully isolated neuroblastoma cells from primary human neuroblastoma tissues by using a magnetic bead-mediated purification system. Since primary neuroblastoma tissues contained CD3- and CD19-positive lymphocytes, total cell suspensions were prepared and incubated with magnetic beads coated with anti-CD3 or with anti-CD19 antibody. After magnetic separation, unbound materials were recovered and analyzed by immunohistochemical staining for NB84, one of the neuroblastoma markers. Immunohistochemical and FACS analyses demonstrated that NB84-positive cells were enriched in the unbound fraction. Subsequently, unbound materials were seeded on cell culture plates and maintained at 37 degrees C overnight. After incubation, non-adherent cells were collected and stained with anti-NB84 antibody. Under our experimental conditions, a significant increase in the number of NB84-positive cells was observed. Furthermore, our purified NB84-positive cells responded to all-trans retinoic acid and nerve growth factor better than the initial primary cells. Collectively, our present results suggest that magnetic bead-mediated purification enriches neuroblastoma cells which retain their biological properties.
Asunto(s)
Técnicas de Cultivo de Célula/métodos , Separación Celular/métodos , Microesferas , Neuroblastoma/patología , Neoplasias del Sistema Nervioso Periférico/patología , Antígenos CD19/análisis , Complejo CD3/análisis , Niño , Preescolar , Femenino , Óxido Ferrosoférrico/química , Citometría de Flujo , Humanos , Inmunohistoquímica , Lactante , Linfocitos/química , Masculino , Células Tumorales CultivadasRESUMEN
Hepatoblastoma is one of the common pediatric solid tumors with frequent mutation of the beta-catenin gene which might be an early event of its carcinogenesis. However, the detailed molecular mechanism is still unknown. We studied the expression levels of CCAAT/enhancer binding protein alpha (C/EBPalpha) and C/EBPbeta, which regulate differentiation and growth of embryonic hepatocytes, to establish whether or not they were involved in affecting the clinical behavior of hepatoblastoma. The quantitative real-time reverse transcriptase-PCR revealed that expression of C/EBPalpha mRNA was significantly up-regulated in tumors 223% (p=0.013) as compared with that in adjacent normal livers, while expression of C/EBPbeta was down-regulated to 27% (p=0.002). Of interest, the immunohistochemical analysis showed that expression of C/EBPalpha was higher and that of C/EBPbeta lower in the poorly differentiated tumor cells than in the well-differentiated cells within the same tumor. Furthermore, high expression of C/EBPalpha (p=0.047) as well as low expression of C/EBPbeta (p=0.025) was significantly associated with poor prognosis of the patients. Cox hazard model suggested that expression of C/EBPalpha and that of C/EBPbeta were independent indicators to predict the prognosis from age but not from histology. Thus, expression of C/EBP proteins may play an important role in the genesis and clinical behavior of hepatoblastoma probably by inducing different stages of arrest of differentiation.
Asunto(s)
Proteína alfa Potenciadora de Unión a CCAAT/biosíntesis , Proteína beta Potenciadora de Unión a CCAAT/biosíntesis , Regulación Neoplásica de la Expresión Génica , Hepatoblastoma/metabolismo , Neoplasias Hepáticas/metabolismo , Proteína alfa Potenciadora de Unión a CCAAT/fisiología , Proteína beta Potenciadora de Unión a CCAAT/fisiología , Diferenciación Celular , Proliferación Celular , Regulación hacia Abajo , Hepatoblastoma/patología , Humanos , Inmunohistoquímica , Neoplasias Hepáticas/patología , Pronóstico , Modelos de Riesgos Proporcionales , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Regulación hacia ArribaRESUMEN
BACKGROUND: We demonstrate the long-term effectiveness of KRP-203 treatment in combination with a subtherapeutic dose of cyclosporine A (CsA) on rat renal allografts. METHODS: We tested the effect of KRP-203 in combination with CsA using a rat skin allograft model. The Pharmacokinetic interaction between CsA and KRP-203 was evaluated. The selectivity of KRP-203 for sphingosine-1-phosphate (S1P)1 and S1P3 receptors were investigated in vitro. Heart rate alteration following bolus injection of phosphorylated KRP-203 (KRP-203-P) or FTY720 (FTY720-P) was also monitored in rats. Finally, the long-term effectiveness of KRP-203 in conjunction with a low dose of CsA was investigated in a rat renal transplantation model. RESULTS: Administration of KRP-203 with CsA prolonged skin allograft survival. KRP-203 and CsA had no effect on the pharmacokinetics of the other. While FTY720-P activated both S1P1 and S1P3 receptors, KRP-203-P selectively activated S1P1, but not the S1P3 receptor (EC50:>1000 nM). Compared to FTY720-P, a tenfold higher dose of KRP-203-P was necessary to induce transient bradycardia. With a low dose of CsA (1 mg/kg/day), KRP-203 (0.3 mg/kg/day) significantly prolonged renal allograft survival (P<0.05, survival time: 9.8 days (CsA) vs. >27.4 days (CsA+KRP)). Although a higher dose of CsA (3 mg/kg/day) alone kept recipients alive, this caused severe renal graft dysfunction. Use of KRP-203 (3 mg/kg/day) in conjunction with CsA markedly improved graft function (P<0.05, creatinine clearance: 0.41+/-0.25 ml/min [CsA] vs. 1.15+/-0.16 ml/min [CsA+KRP]). CONCLUSIONS: The selectivity of KRP-203 for S1P1 reduces the risk of bradycardia, and the combination therapy of KRP-203 with CsA represents a safe and effective strategy for use in renal transplantation.
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Ciclosporina/uso terapéutico , Trasplante de Riñón/inmunología , Compuestos de Sulfhidrilo/uso terapéutico , Animales , Señalización del Calcio/fisiología , Quimioterapia Combinada , Supervivencia de Injerto , Frecuencia Cardíaca/efectos de los fármacos , Complejo Mayor de Histocompatibilidad , Masculino , Ratas , Ratas Endogámicas F344 , Receptores de Lisoesfingolípidos/agonistas , Trasplante HomólogoRESUMEN
Hepatoblastoma is one of the most common malignant liver tumors in young children. Recent evidences have suggested that the abnormalities in Wnt signaling pathway, as seen in frequent mutation of the beta-catenin gene, may play a role in the genesis of hepatoblastoma. However, the precise mechanism to cause the tumor has been elusive. To identify novel hepatoblastoma-related genes for unveiling the molecular mechanism of the tumorigenesis, a large-scale cloning of cDNAs and differential screening of their expression between hepatoblastomas and the corresponding normal livers were performed. We constructed four full-length-enriched cDNA libraries using an oligo-capping method from the primary tissues which included two hepatoblastomas with high levels of alpha-fetoprotein (AFP), a hepatoblastoma without production of AFP, and a normal liver tissue corresponded to the tumor. Among the 10,431 cDNAs randomly picked up and successfully sequenced, 847 (8.1%) were the genes with unknown function. Of interest, the expression profile among the two subsets of hepatoblastoma and a normal liver was extremely different. A semiquantitative RT-PCR analysis showed that 86 out of 1188 genes tested were differentially expressed between hepatoblastomas and the corresponding normal livers, but that only 11 of those were expressed at high levels in the tumors. Notably, PLK1 oncogene was expressed at very high levels in hepatoblastomas as compared to the normal infant's livers. Quantitative real-time RT-PCR analysis for the PLK1 mRNA levels in 74 primary hepatoblastomas and 29 corresponding nontumorous livers indicated that the patients with hepatoblastoma with high expression of PLK1 represented significantly poorer outcome than those with its low expression (5-year survival rate: 55.9 vs 87.0%, respectively, p=0.042), suggesting that the level of PLK1 expression is a novel marker to predict the prognosis of hepatoblastoma. Thus, the differentially expressed genes we have identified may become a useful tool to develop new diagnostic as well as therapeutic strategies of hepatoblastoma.
Asunto(s)
Perfilación de la Expresión Génica , Hepatoblastoma/genética , Neoplasias Hepáticas/genética , Hígado/metabolismo , Oncogenes , Proteínas Quinasas/genética , Proteínas de Ciclo Celular , Biblioteca de Genes , Humanos , Pronóstico , Proteínas Serina-Treonina Quinasas , Proteínas Proto-Oncogénicas , alfa-Fetoproteínas/análisis , Quinasa Tipo Polo 1RESUMEN
BACKGROUND: The effect of graft length on rejection reaction in small bowel transplantation (SBT), which is poorly understood, is tested using rat allogenic SBT models with a short course of tacrolimus. MATERIALS AND METHODS: Inbred Brown Norway rats (major histocompatibility complex: RT1) and Lewis rats (RT1) were used as donors and recipients, respectively. The intestinal tract of the recipient was partially or totally replaced by segmental (15 cm) or whole (70 cm) donor intestine, using two different SBT models. With tacrolimus treatment (0.64 mg/kg per day, 0-13 postoperative days, intramuscularly), recipients' body weights and their survival were evaluated. To compare the extent of peripheral chimerism, donor passenger leukocytes were followed using flow cytometry with a donor-specific monoclonal antibody, OX-27. For the periodical histologic analysis, heterotopic SBT and protocol biopsies of the graft were also performed with short or long intestinal grafts. RESULTS: In a classical Monchik and Russell orthotopic SBT model, whole SBT recipients survived more than 60 days. However, all of the allogenic segmental SBT recipients died within 14 days without histologic sign of graft rejection. In the modified orthotopic SBT model using a cuff technique without systemic clamping, all recipients with segmental allograft survived longer than 29 days. However, recipients with whole graft tended to survive longer than those with segmental graft. The suffering period, lasting from the onset of rejection to death, was significantly shorter in the segmental group than in the whole group. Flow cytometric analysis showed that recipients with whole intestinal grafts had significantly higher ratio of donor passenger leukocytes in peripheral blood. Histologic studies of the protocol biopsies showed that the shorter graft tended to be more severely rejected than the longer graft. CONCLUSIONS: We have demonstrated experimentally that long intestinal grafts have immunologic advantage over short grafts.
Asunto(s)
Supervivencia de Injerto/inmunología , Intestino Delgado/trasplante , Trasplante Homólogo/inmunología , Animales , Inmunosupresores/uso terapéutico , Intestino Delgado/inmunología , Complejo Mayor de Histocompatibilidad , Ratas , Ratas Endogámicas BN , Ratas Endogámicas Lew , Tacrolimus/uso terapéutico , Trasplante Heterotópico/inmunologíaRESUMEN
We describe an 8-year-old male who had an intrapulmonary solitary fibrous tumor (SFT). SFTs of the pleura are now thought to originate from subpleural mesenchymal cells, and a pathological diagnosis is obtained by a specific marker, i.e., CD34 expression on the tumor cells. The SFT reported here is an extremely rare phenotype in two respects: it originated in a child under age 10 years, and it expanded in an intrapulmonary fashion.
Asunto(s)
Carcinoma/diagnóstico , Neoplasias de Tejido Fibroso/diagnóstico , Neoplasias Pleurales/diagnóstico , Carcinoma/cirugía , Niño , Estudios de Seguimiento , Humanos , Masculino , Neoplasias de Tejido Fibroso/cirugía , Pleura/patología , Neoplasias Pleurales/cirugía , Radiografía Torácica , Resultado del TratamientoRESUMEN
BACKGROUND: Infection is a serious complication after liver transplantation. Immunization is one means of controlling infections. The objective of this study was to investigate the efficacy and safety of simultaneous administration of several vaccines before transplantation and the efficacy and safety of administration under immunosuppressive conditions after transplantation. METHODS: Fifty-eight patients who underwent living-related liver transplantation between April 1994 and March 2000 were included in this study. Simultaneous administration of a maximum of six vaccines was performed in a short period of time before transplantation. We also readministered vaccines to 15 patients with waning antibody titers after transplantation from June 1999. We investigated whether patients could seroconvert for measles, rubella, mumps, and varicella after immunization and how long antibody titers could be retained by measuring them several times throughout the period before and after transplantation. We also examined side effects caused by immunization. RESULTS: The rates of seroconversion against measles, rubella, mumps, and varicella after the pretransplantation vaccination were 82%, 100%, 90%, and 95%, respectively. The rates of reseroconversion against measles, rubella, mumps, and varicella after the posttransplantation revaccination were 85%, 100%, 100%, and 71%, respectively. Although antibody titers against these viruses generally waned with time, no patient exhibited any serious illness or side effects. CONCLUSION: Although 12 of 58 patients (21%) had an infection, pretransplantation immunization was effective to prevent serious illness, especially for the 6 months after transplantation. Posttransplantation live-vaccine administration under immunosuppressive conditions is effective and safe.
Asunto(s)
Inmunización , Trasplante de Hígado , Adolescente , Anticuerpos Antivirales/sangre , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Vacunas Virales/efectos adversos , Vacunas Virales/inmunologíaRESUMEN
Expression of the p16INK4A (p16), p15INK4B (p15), and p14ARF genes, located at 9p21, was examined in pediatric neuroblastoma (NB), Ewing's sarcoma (ES), and rhabdomyosarcoma (RMS). p16 expression was absent in 4 of 5 ESs, and 2 of these 4 cases died. p16 expression was reduced or absent in 10 of 12 RMSs, and 4 of these 10 cases died. These results suggested the possibility that p16 expression was associated with the progression of ES and RMS. There has been no previous report on p14ARF in NB. Our investigation might indicate that abnormal expression of the p16 and p14ARF was associated with a poor prognosis in NB, although in some cases of NB normal p16 and abnormal p14ARF expression was seen. These findings suggest an important role of p14ARF gene in the tumorigenesis of NB. The different incidence of expression of the p16, p15, and p14ARF genes in these 3 tumor types may reflect differences of the molecular process through which the 3 tumors develop. Our results suggest that abnormal expression of the p16 and/or p14ARF may be associated with a poor prognosis in these 3 tumors.
Asunto(s)
Proteínas de Ciclo Celular/genética , Aberraciones Cromosómicas , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Neoplasias/metabolismo , Proteína p14ARF Supresora de Tumor/genética , Proteínas Supresoras de Tumor/genética , Southern Blotting , Western Blotting , Línea Celular Tumoral , Inhibidor p15 de las Quinasas Dependientes de la Ciclina , ADN/química , Metilación de ADN , Eliminación de Gen , Humanos , Neuroblastoma/genética , Neuroblastoma/mortalidad , Pronóstico , ARN/química , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Rabdomiosarcoma/genética , Rabdomiosarcoma/mortalidad , Sarcoma de Ewing/genética , Sarcoma de Ewing/mortalidadRESUMEN
The benign epithelial neoplasm liver cell adenoma is rare, especially in childhood. We report 2 such cases, 1 of which was associated with Prader-Willi syndrome. Differential diagnosis of the liver cell adenomas on the basis of histopathologic findings proved difficult and was based on the absence of cellular and nuclear atypia, mitotic activity, and invasive growth. In both cases, immunohistochemical staining demonstrated the nuclear accumulation of beta-catenin, and in 1 case, the tumor cells carried a mutation of the beta-catenin gene. Recently, disregulation of the Wnt/beta-catenin pathway, attributable to abnormalities of the beta-catenin gene, has been reported to be a major event in the development of hepatocellular carcinomas and hepatoblastomas. Our report may be the first to describe the beta-catenin abnormalities in childhood liver cell adenoma. These findings imply that abnormalities of beta-catenin can be an early initiating event in human liver tumorigenesis.
Asunto(s)
Adenoma de Células Hepáticas/química , Proteínas del Citoesqueleto/análisis , Neoplasias Hepáticas/química , Transactivadores/análisis , Adenoma de Células Hepáticas/genética , Adenoma de Células Hepáticas/patología , Adolescente , Secuencia de Bases , Núcleo Celular/química , Niño , Proteínas del Citoesqueleto/genética , Femenino , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patología , Mutación , Reacción en Cadena de la Polimerasa , Transactivadores/genética , beta CateninaRESUMEN
BACKGROUND: Intestinal grafts greatly affect nutrition and immunology in the host. The growth of the recipient and incidence of graft-versus-host disease depend on graft length. A larger graft may affect the host immune system, but little is known about how the length of the intestinal graft severely affects surgical intervention. We developed a cervical small bowel transplantation (SBT) rat model that minimized technical variations using a cuff method and studied the effects of graft length on surgical damage in SBT. MATERIALS AND METHODS: We transplanted a whole (70 cm) or partial (15 cm) intestine into a syngeneic rat combination of LEW (MHC haplotype: RT1(l)) to LEW and evaluated changes in perioperative hemodynamics and the endogenous endotoxin level. Natural killer (NK) cell activity in the peripheral blood and the immunologic response of the recipient spleen were also studied. RESULTS: In the whole SBT model, body weight loss was more severe than in the segmental SBT model; the rats in the former model often died, while all in the latter survived indefinitely. The systemic blood pressure markedly decreased in the whole SBT group immediately after reperfusion. The proliferative activity of splenic lymphocytes stimulated by concanavalin A was also more severely inhibited in the former model than in the latter postoperatively. NK cell activity in the whole SBT rats declined more severely than the segmental SBT rats 3 days postoperatively. CONCLUSION: The longer graft severely induced surgical intervention; and influenced host immunosuppression, resulting in the higher mortality in rats undergoing whole SBT.
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Intestino Delgado/trasplante , Inmunología del Trasplante , Trasplante/efectos adversos , Trasplante/métodos , Animales , Presión Sanguínea/fisiología , Peso Corporal , Endotoxinas/metabolismo , Células Asesinas Naturales/inmunología , Activación de Linfocitos/inmunología , Ratas , Inmunología del Trasplante/fisiologíaRESUMEN
Ras guanyl nucleotide-releasing protein 2 (RASGRP2), one of the Ras guanine exchange factors, is implicated as a critical regulator of inside-out integrin activation in human lymphocytes, neutrophils and platelets. However, the activities of this protein in endothelial cells remain unclear. In the current study, we identify a physiological function in blood vessel formation for XRASGRP2, which is the Xenopus ortholog of mammalian RASGRP2. XRASGRP2 over-expression induced ectopic vascular formation, and XRASGRP2-knockdown embryos showed delayed vascular development. We also investigated the upstream signaling of XRASGRP2 in endothelium formation. XRASGRP2 expression was up-regulated in the presence of VEGF-A and down-regulated following VEGF-A depletion. XRASGRP2 knockdown abolished the ectopic induction of endothelial cells by VEGF-A in the posterior ventral blood island. These results suggest that XRASGRP2 is essential for vascular formation during Xenopus development.
Asunto(s)
Vasos Sanguíneos/metabolismo , Proteínas de Xenopus/fisiología , Xenopus/metabolismo , Factores de Intercambio de Guanina Nucleótido ras/fisiología , Animales , Diferenciación Celular/genética , Células Endoteliales/metabolismo , Factores de Intercambio de Guanina Nucleótido/genética , Factores de Intercambio de Guanina Nucleótido/metabolismo , Humanos , Transducción de Señal/fisiología , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , Xenopus/genética , Proteínas de Xenopus/genética , Factores de Intercambio de Guanina Nucleótido ras/genéticaRESUMEN
Body stalk anomaly is characterized by severe scoliosis, severe pulmonary hypoplasia, and giant omphalocele. The prognosis of the disease is poor and most obstetricians consider it fatal. Very few patients with body stalk anomaly survive. We report the case of a baby diagnosed with body stalk anomaly in fetal life, who was saved by intensive care after birth. We closed the giant omphalocele successfully by placing karaya gum sheets over it, which created a humidified environment and promoted natural skin epithelization over the skin defect.
Asunto(s)
Anomalías Múltiples/terapia , Hernia Umbilical/cirugía , Pulmón/anomalías , Escoliosis/congénito , Adulto , Femenino , Humanos , Recién Nacido , Sobrevivientes , SíndromeRESUMEN
The Protractor is a self-retaining ring retractor used mainly for minilaparotomies in adults. We report our positive results of using this retractor in pediatric surgery. We performed surgery with the aid of the Protractor in 57 pediatric patients aged from 1 day old to 16 years old. The Protractor allowed a wide operative view and did not cause any major complications. It was especially useful for Kasai's portoenterostomies and ureteroneocystostomy (Cohen's repair). In neonatal surgery, the Protractor not only provided a wide operative view, but also prevented the washing fluid from overflowing onto the covering sheet. In appendectomy, the Protractor protected the surgical wound from contaminated ascites and the appendix. The Protractor is a very useful tool for assisting with various abdominal operations in infants and children.
Asunto(s)
Laparotomía/instrumentación , Pediatría/instrumentación , Instrumentos Quirúrgicos , Adolescente , Apendicectomía/instrumentación , Niño , Preescolar , Cistostomía/instrumentación , Diseño de Equipo , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Ureterostomía/instrumentaciónRESUMEN
We report an unusual case of epigastric heteropagus in a female neonate. The parasite had a single lung as well as a liver, stomach, intestine, pancreas, ovaries, a single kidney, and a bladder. These visceral organs were located in the abdominal and pelvic space of the autosite, and the pelvic bone and lower legs of the parasite were fused with the respective organs in the autosite. The infant underwent operations to remove the central leg and some of the intra-abdominal organs of the parasite, but she must still undergo further surgery to remove the genitourinary organs and pelvic bones of the parasite, and repair her very abducted and rotated right leg.
Asunto(s)
Anomalías Múltiples/diagnóstico , Enfermedades en Gemelos , Gemelos Siameses/cirugía , Anomalías Múltiples/cirugía , Femenino , Estudios de Seguimiento , Tracto Gastrointestinal/anomalías , Humanos , Recién Nacido , Riñón/anomalías , Pierna/anomalías , Pulmón/anomalías , Ovario/anomalías , Tomografía Computarizada por Rayos X , Vejiga Urinaria/anomalíasRESUMEN
BACKGROUND: Replacement of calcineurin inhibitor (CI) with anti-metabolic agents in transplant patients with CI-induced nephrotoxicity is performed clinically and improves renal function, but increases the risk of rejection. We investigated whether the change from cyclosporine (CsA) to a limited dose of mycophenolic acid (MPA) together with a new sphingosine-1-phosphate (S1P) receptor agonist, KRP-203, is sufficient to prevent both transplant vasculopathy and CsA-induced nephrotoxicity. METHODS: Orthotopic aortic transplantation was conducted in a high-responder rat combination of Dark Agouti (DA; major histocompatibility complex [MHC] haplotype RT-1a) to Lewis (RT-1(l)). After CsA administration (15 mg/kg/day) for 2 weeks, the recipients were divided into the following treatment groups for 6 weeks: MPA (10 mg/kg); KRP-203 (KRP; 1 mg/kg); and MPA + KRP. Serum creatinine (Cr), arteriolar hyalinosis and expression of transforming growth factor (TGF)-beta1 in the recipient kidney were examined as parameters indicating nephrotoxicity. Intimal hyperplasia was assessed by vascular occlusion, and graft-infiltrated cells were semi-quantitatively evaluated histologically and then characterized immunohistochemically. RESULTS: Continuous CsA treatment attenuated intimal hyperplasia and cell infiltration (2.9 +/- 0.3% and 0.4 +/- 0.1; p < 0.01 vs vehicle), but increased Cr and hyalinosis (0.43 +/- 0.03 mg/dl and 57.2 +/- 0.4%; p < 0.01) with upregulated TGF-beta1. Replacement of CsA by MPA or KRP treatment alone improved nephrotoxicity, but worsened intimal hyperplasia and cell infiltration. Conversion to MPA + KRP treatment prevented nephrotoxicity (Cr, 0.32 +/- 0.02 mg/dl; hyalinosis, 5.6 +/- 1.3%; p < 0.01 vs CsA) and markedly suppressed intimal hyperplasia and cell infiltration (3.6 +/- 1.2% and 1.0 +/- 0.3; p = not significant vs CsA), with reduced T-cell infiltrates in the graft. CONCLUSIONS: Changing from CsA to a combined therapy of MMF with S1P agonist is a promising strategy in clinical transplantation to overcome CI-induced nephrotoxicity and chronic rejection.
Asunto(s)
Aorta/trasplante , Enfermedades de la Aorta/prevención & control , Ciclosporina/uso terapéutico , Inmunosupresores/uso terapéutico , Enfermedades Renales/prevención & control , Ácido Micofenólico/uso terapéutico , Receptores de Lisoesfingolípidos/agonistas , Compuestos de Sulfhidrilo/uso terapéutico , Animales , Aorta/metabolismo , Aorta/patología , Enfermedades de la Aorta/epidemiología , Arteriopatías Oclusivas/epidemiología , Arteriopatías Oclusivas/prevención & control , Recuento de Células Sanguíneas , Ciclosporina/efectos adversos , Quimioterapia Combinada , Hiperplasia/prevención & control , Inmunohistoquímica , Inmunosupresores/efectos adversos , Enfermedades Renales/inducido químicamente , Enfermedades Renales/etiología , Macrófagos/patología , Masculino , Ratas , Ratas Endogámicas , Retratamiento , Linfocitos T/patología , Trasplante Homólogo , Túnica Íntima/patologíaRESUMEN
OBJECTIVE: Based on development of stem cell technology, newborn tissue, even undergoing cryopreservation, possesses promising potential as a donor source in the field of organ transplantation. However, the precise regeneration processes remains unclear. This study was designed to investigate the regenerative potential of newborn intestine with or without cryopreservation in the transplantation. METHODS: Newborn rat intestines with or without cryopreservation were transplanted subcutaneously into the syngeneic host, and specimens were evaluated by histology, multiple immunostaining, and comprehensive gene expression analysis. RESULTS: We determined that newborn rat intestine possessed regenerative potential in the syngeneic host even after cryopreservation, where angiogenesis was induced early in the submucosa with subsequent maturation in the crypts. Furthermore, newborn intestinal graft could facilitate the survival of maturation-incompetent 10-day-old graft that lacked regenerating activity (P < 0.01, n = 13). Tissue aggregates from the maturation-incompetent graft underwent reconstitution of their histologic configuration in the presence of newborn intestinal aggregates. Comprehensive gene expression analysis showed that 37 genes were preferentially up-regulated, while 19 genes were down-regulated in the regenerating 10-day-old graft (supported by the newborn graft). CONCLUSIONS: Regeneration of newborn intestine is implicated in neo-angiogenesis in the host, and the newborn intestinal graft is capable of mediating the survival of the maturation-incompetent 10-day-old graft. Notwithstanding ethical and legal limitations in the clinic, these results may provide new insights into the regenerative role of newborn grafts.
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Intestino Delgado/fisiología , Intestino Delgado/trasplante , Regeneración/fisiología , Inmunología del Trasplante , Animales , Animales Recién Nacidos , Biopsia con Aguja , Criopreservación/métodos , Femenino , Rechazo de Injerto , Supervivencia de Injerto , Inmunohistoquímica , Intestino Delgado/patología , Ratas , Ratas Endogámicas Lew , Factores de Riesgo , Sensibilidad y Especificidad , Factores de Tiempo , Trasplante de Tejidos/métodos , Trasplante HomólogoRESUMEN
Bronchial asthma is an increasingly common disorder that remains poorly understood and difficult to manage. The disease is characterized by airway hyperresponsiveness, chronic inflammation, and mucus overproduction. Based on the finding that leukotriene B4 receptor 1 (BLT1) is expressed highly in Th2 lymphocytes, we analyzed the roles of BLT1 using an OVA-induced bronchial asthma model. BLT1-null mice did not develop airway hyperresponsiveness, eosinophilic inflammation, and hyperplasia of goblet cells. Attenuated symptoms were accompanied by reduced IgE production, and accumulation of IL-5 and IL-13 in bronchoalveolar lavage fluid, suggesting attenuated Th2-type immune response in BLT1-null mice. Peribronchial lymph node cells of sensitized BLT1-null mice showed much attenuated proliferation and production of Th2 cytokines upon re-stimulation with Ag in vitro. Thus, LTB4-BLT1 axis is required for the development of Th2-type immune response, and blockade of LTB4 functions through BLT1 would be novel and useful in the effort to ameliorate bronchial asthma and related Th2-biased immune disorders.
Asunto(s)
Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/inmunología , Leucotrieno B4/fisiología , Receptores de Leucotrieno B4/deficiencia , Sistema Respiratorio/inmunología , Células Th2/inmunología , Animales , Asma/genética , Asma/inmunología , Asma/metabolismo , Calcio/metabolismo , Modelos Animales de Enfermedad , Inmunidad Innata/genética , Inmunoglobulina E/biosíntesis , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Noqueados , Peroxidasa/metabolismo , Receptores de Leucotrieno B4/genética , Receptores de Leucotrieno B4/fisiología , Sistema Respiratorio/metabolismo , Células Th2/metabolismoRESUMEN
Intrapericardial diaphragmatic hernia is a very rare phenotype of neonatal diaphragmatic hernia which is thought to be caused by the developmental failure of the septum transversum. There have been only 10 cases reported since 1980, and among them, only 2 cases were diagnosed in fetal life. We herein report a new case that was diagnosed in fetal life, and pericardiocentesis was performed at 27 weeks of gestation. This is the first case to undergo a fetal interventional therapy. After birth, the patient successfully underwent closure of the hernia, despite severe pulmonary hypoplasia.
Asunto(s)
Feto/cirugía , Hernia Diafragmática/diagnóstico , Derrame Pericárdico/etiología , Pericardiocentesis/métodos , Pericardio/patología , Diagnóstico Prenatal , Adulto , Diagnóstico Diferencial , Femenino , Hernia Diafragmática/complicaciones , Hernia Diafragmática/cirugía , Humanos , Recién Nacido , Fenotipo , EmbarazoRESUMEN
BACKGROUND: The characteristics of adenovirus-mediated gene transfer into the kidney are not well examined. We studied the effects of contact time and temperature on adenovirus-mediated transgene expression in rat kidneys, using catheter-based in vivo gene transfer and a rat renal transplant model ex vivo. METHODS: An adenovirus vector containing the luciferase (Ad-Luc) or beta-galactosidase (Ad-LacZ) gene was introduced in vivo into the kidney via a renal artery catheter. Various contact times and temperatures were evaluated. Ex vivo, the renal graft was injected with Ad-Luc through the renal artery, chilled for 60 min and then transplanted. Luciferase expression was evaluated periodically by a non-invasive bioimaging system or histology. Cells expressing the LacZ gene were identified by immunoelectron microscopy. RESULTS: In in vivo gene transfer, successful transgene expression was achieved; however, its efficiency was independent of contact time or temperature. In ex vivo gene transfer, transgene expression in the renal graft peaked early and gradually decreased. Strong gene expression was observed in the recipients' livers. LacZ expression was detected in fibroblasts, parietal epithelial cells of Bowman's capsule, mesangial cells, podocytes and tubular cells. CONCLUSIONS: This study generated new information about in vivo and ex vivo gene transfer into the kidney, which would be useful for renal gene therapy.
Asunto(s)
Adenoviridae , Técnicas de Transferencia de Gen , Vectores Genéticos , Riñón/fisiología , Operón Lac/fisiología , Luciferasas de Luciérnaga/metabolismo , Animales , Expresión Génica/fisiología , Masculino , Ratas , Ratas Endogámicas Lew , Temperatura , Factores de Tiempo , Transgenes/fisiologíaRESUMEN
The theory of gastrointestinal duplication and enteric cyst embryogenesis was verified by examining the developmental process of this experimentally induced anomaly. In Cynopus pyrrhogaster (amphibian) embryos (stage 18), the dorsal midline structures (including the neural plate and notochord) were split regionally to induce partial separation of the notochord and gut anlage endoderm herniation between the split elements of the notochord. Following this procedure, the embryonic development was traced morphologically and histologically. Control embryos were cultured without the procedure. Following the incubation and breeding period, gastrointestinal duplication and enteric cysts were observed with vertebral anomaly, spina bifida, split cord malformation and subcutaneous manifestations in the mature animals. The combination of anomalies that was observed in these experimental animals is consistent with that found in "split notochord syndrome." No abnormal morphology or histology was observed in the control group. The embryogenetic theory of gastrointestinal duplication and enteric cysts was thus verified by simulating the partial separation of the notochord, which induced split notochord syndrome in laboratory animals. The results indicate that gastrointestinal duplication and enteric cysts may arise through a process of herniation of the gut anlage endoderm between split elements of the notochord.