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Objective. The purpose of this study was to determine how well liver position, the lung area-to-head circumference (L/H) ratio, and the lung-to-thorax transverse area (L/T) ratio predicted the need for extracorporeal membrane oxygenation (ECMO) and survival in fetuses with isolated congenital diaphragmatic hernia (CDH). Methods. Antenatal records of 25 fetuses with isolated left-sided CDH who were born by cesarean delivery under fetal stabilization at this institution were reviewed. The latest determinations of the L/H and L/T ratios before birth (between 34 and 38 weeks’ gestation) were compared on the basis of the cutoff points for mortality: less than 1.0 versus 1.0 or greater for the L/H ratio and 0.08 or less versus greater than 0.08 for the L/T ratio. Outcome measures assessed were survival (discharge to home) and the need for ECMO. Results. Overall survival was 64% (16/25). Postnatal survival in fetuses with an L/T ratio of 0.08 or less was statistically lower than in those with an L/T ratio of greater than 0.08 (33% versus 81%; P = .0308). The percentage requiring ECMO in the group with an L/T ratio of 0.08 or less was also higher than that of the group with an L/T ratio of greater than 0.08, but the difference was not statistically significant (67% versus 25%; P = .0872). Neither the L/H ratio nor herniation of the fetal liver into the chest affected survival or the need for ECMO. Conclusions. In fetuses with isolated CDH at term or near term, the L/T ratio may be a better predictor of outcome than the L/H ratio or liver herniation. Key words: congenital diaphragmatic hernia; lung area-to-head circumference ratio; lung hypoplasia; lung-to-thorax transverse area ratio; sonography.

We previously showed that fetal stabilization (FS) could improve the prognosis of congenital diaphragmatic hernia (CDH) patients. The aim of this study is to elucidate the effect of FS in normal neonatal rats. Pregnant Sprague-Dawley rats were treated by experimental protocols on day 21 of gestation. In the FS-group, they received morphine hydrochloride via the placenta before undergoing a caesarean section. In the control group (C-group), they received no morphine hydrochloride. All neonatal rats were managed under mechanical ventilation. We collected the blood samples and bronchoalveolar lavage fluid (BALF) at birth and at 4 h after birth in both groups and the cytokine levels in those samples were measured. The specimens obtained from the right lung were stained with anti-TNF-alpha antibody. The levels of serum TNF-alpha at birth and IL-6 at 4 h after birth in the FS-group decreased, in comparison to those in the C-group. The staining intensity of anti-TNF-alpha antibody in the FS-group was weaker than that in the C-group. FS reduced the production of inflammatory cytokines on neonatal rats, which was controlled by mechanical ventilation. This effect may beneficially reduce the occurrence of persistent pulmonary hypertension of neonate (PPHN), which is induced by stress in CDH patients.

We demonstrated previously that the additive-type recombinant Sendai virus (rSeV) is highly efficient for use in pulmonary gene transfer; however, rSeV exhibits inflammatory responses. To overcome this problem, we tested newly developed non-transmissible constructs, namely, temperature-sensitive F-deleted vector, rSeV/dF (ts-rSeV/dF) and a rSeV with all the envelope-related genes deleted (rSeV/dFdMdHN), for pulmonary gene transfer in neonatal mice, by assessing their toxicity and immune responses. The gene expression in the lungs of neonatal ICR mice peaked on day 2, then gradually decreased until almost disappearing at 14 days after infection in all constructs. Loss of body weight and mortality rate, however, were dramatically improved in mice treated with SeV/dFdMdHN (mortality=0%, n=41) and ts-rSeV/dF (24.2%, n=33) compared with additive rSeV (70.7%, n=58). Although the deletion of envelope-related genes of SeV had a small impact on the production of antibody and cytotoxic T-lymphocyte activity in both adults and neonates, a dramatic reduction was found in the events related to innate responses, including the production of proinflammatory cytokines, particularly in the case of neonates. These results indicate that pulmonary gene transfer using SeV/dFdMdHN warrants further investigation for its possible use in developing safer therapeutics for neonatal lung diseases, including cystic fibrosis.

Background/Purpose: Cloacal malformations are rare anomalies, occurring in females, and in which they demonstrate a single perineal orifice for urethra, vagina, and rectum. Prenatal ultrasonograms (US) of cloacal malformations sometimes show ascites, hydrocolpos, and hydronephrosis. We herein describe the characteristic prenatal US and magnetic resonance imaging (MRI) findings of the cloacal malformations associated with meconium peritonitis. Methods: The pre- and postnatal records of 11 newborn patients with cloacal malformations, treated in our hospital from 1988 to 2004, were reviewed. All fetuses underwent prenatal US by experienced obstetricians, whereas in addition, fetal MRI was performed in 1 patient. Results: The prenatal US and/or MRI findings showed fetal ascites, a multicystic pelvic mass, oligohydramnios, and bilateral hydronephrosis in 5 of 11 patients with cloacal malformations. In these 5 cases, postnatal examinations showed associated hydrocolpos, hydrometrocolpos, and bilateral hydronephrosis; furthermore, 4 of these 5 cases also showed meconium peritonitis at laparotomy. Conclusions: The prenatal US and MRI findings, showing fetal ascites, multicystic pelvic mass, bilateral hydronephrosis, and oligohydraminios are highly suggestive of the cloacal malformations associated with meconium peritonitis.

Background/purpose Recently, valine, which is one of the branched chain amino acids, has been reported to enhance liver regeneration after hepatectomy in the rat. The aim of this study was to investigate the effect of enteral valine supplementation on intestinal adaptation. Materials/methods Seven-week-old male Lewis rats underwent a 90% small bowel resection. The rats were randomly divided into two groups: group V (valine-rich diet) and group S (standard rat chow), according to the diet. The rats were sacrificed at the operation day and on postoperative days (POD) 7, 14, 30, and 60. The metrics were body weight (BW), blood amino acids, urine organic acids, and morphology of the residual small intestine. Results The BW and the intestinal wet weight, jejunal crypt depth, and proliferating cell nuclear antigen–positive cells in group V at POD 7 were significantly higher than those values in group S, while those in group V at POD 30 and 60 were smaller than those in group S. The urine methylmalonic acid (MMA) level in group V at POD 30 and 60 was much higher than in group S. Conclusion Valine enhanced intestinal adaptation after massive small bowel resection in the acute phase. However, the long-term supplementation disturbed intestinal adaptation, which might be due to the high production of MMA.

MYCN is the most powerful prognostic factor in cases of older children. However, how MYCN is related to the prognosis of infantile cases is not clear. A mass screening program was carried out by measuring urinary catecholamine metabolites (VMA and HVA) from 6-month-old infants. Of 2084 cases detected by the screening program, MYCN amplification (MNA) was examined by Southern blot analyses in 1533 cases from 1987 to 2000. Of the 1533 cases examined, 1500 (97.8%) showed no MNA, 20 cases (1.3%) showed MNA from three to nine copies, and 13 (0.8%) cases showed more than 10 copies. The 4-year overall survival rates of these three groups (99, 89 and 53%, respectively) were significantly different (P

Background/Purpose To select the optimal treatment according to the degree of malignancy of neuroblastoma, it is essential to accurately and rapidly identify any genetic abnormalities associated with the prognosis. This study aims to assess the correlation between the combination of prognostic factors and the biologic findings of neuroblastoma using a highly sensitive analysis of prognostic factors. Methods In 44 neuroblastoma primary samples, we determined the gene dosages of MYCN and Survivin (as the target of 17q gain) and the expression levels of MYCN , Survivin , and BIN1 using highly sensitive analysis (the quantitative polymerase chain reaction method); furthermore, we assessed the correlation between the combination of their prognostic factors and the biology of neuroblastoma. Results The gene dosage of MYCN or Survivin was significantly associated with all known prognostic factors. The expression level of MYCN or Survivin was not significantly associated with any prognostic factors, whereas the expression level of BIN1 was significantly associated with 5 of 6 prognostic factors. Regarding the combination of MYCN amplification and 17q gain (the gene dosage of Survivin ), and the low expression of BIN1 , the rates of advanced stages (stage III or IV) were 100% for the cases with 3 factors, 63% for the cases with 2 factors, 42% for the cases with 1 factor, and 0% for the cases with null factor. Furthermore, the survival rates were 20% for the cases with 3 factors, 50% for the cases with 2 factors, 100% for the cases with 1 factor, and 100% for the cases with null factor. Conclusion The combination of gene dosages of MYCN and Survivin and the expression level of BIN1 using the quantitative polymerase chain reaction method was significantly correlated with the clinical stage and the patients' outcome. This combination of biologic factors may enhance the accuracy to the conventional criteria, but this would have to be shown in a much larger study that is adequately powered to detect such an advantage.

The placement of the Hickman catheter in the central veins is thought to be an effective method for providing venous access in various clinical situations in children. The catheter is usually inserted by the percutaneous approach, but in some cases various troublesome complications can occur, such as sheath introducer kinking or damage, in addition to other major ones. Therefore, some modified techniques, using vascular dilators, both to dilate the route and to avoid such complications, have been developed and investigated to obtain a smooth and safe percutaneous insertion of the Hickman catheter in children. A total of 41 Hickman catheters were inserted by the percutaneous method in 41 pediatric patients from 1996 to 2004 in our department. Sixteen catheters were inserted by means of a standard method, using the manufacturer's insertion kit, and 25 catheters were inserted by means of a modified method, namely, using various sized vascular dilators. The length of time for the procedure, the complication rate, and the changes in the serum C-reactive reaction (CRP) levels were then compared between the standard and the modified methods. Those parameters were also compared between a right-side and left-side approach using both methods, to clarify which side was better for the insertion of this catheter. The length of time for the catheter replacement procedure in the standard group was significantly longer than that in modified one. The occurrence rate for both the kinking and small damage to the sheath introducer in the standard group was higher than that in the modified one. The peak of serum CRP in the modified group was significantly lower than that in the standard one. When comparing a right-side and left-side approach, 7 catheters out of 16 were inserted by the right-side approach in the standard group, while 10 catheters out of 25 were done by the right-side approach in the modified group. The length of time for the procedure for the left-side approach was significantly shorter than that for the right-side one in both groups. No difference in technical complications was observed between the two different approaches in the modified group, while complications when using the right-side approach often occurred in the standard group. The peak of serum CRP in the left-side approach was lower than that in the right-side one in both groups. The use of the modified percutaneous method, using various sized vascular dilators and the left-side approach, was therefore found to be useful for the safe and smooth placement of the Hickman catheter in children.

But: II n'y a aucune etude nationale pour les patients avec un rhabdomyosarcome au Japon. Le but de cette etude est d'analyser l'etat actuel du traitement et du devenir de ces rhabdomyosarcomes. Patients et Methodes: De 1982 a 1996, 79 rhabdomyosarcomes etaient enregistres par ce groupe d'etude pour les tumeurs solides malignes dans la zone de Kyushu. Les facteurs pronostiques et les traitements etaient analyses, bases sur la survie a 5 ans. Le staging etait fait en accord avec la classification du groupe rhabdomyosarcome (IRS). Resultats: La survie a 5 ans pour tous les patients etait de 39,1 %. Le taux de survie pour chaque facteur etait le suivant: 1) rapport avec le groupe: 77,8% pour le groupe I, 51,9% pour le groupe II, 33,7% pour le groupe III et 20,2% pour le groupe IV; 2) en fonction de la localisation initiale: 56,3% pour tete et cou, 43,8% pour le tronc, 12,5% pour les extremites, 58,3% pour le pelvis et la localisation genito-urinaire et 30,5% pour les autres; 3) histologie: 35,8% etaient de type embryonnaire, 36,8% etaient de type alveolaire. Conclusion: Quoique le devenir de cette etude soit mediocre, pour ameliorer le devenir, une nouvelle etude nationale pour le rhabdomyosarcome doit etre realisee au Japon.

Purpose Phthalate esters have been shown to cause undescended testes (UT) in rats. It has been proposed that Leydig insulin-like factor 3 (INSL3) may play an important role in testicular descent. The aim of this study was to investigate the effect of mono-n-butyl phthalate (MBP) on both the INSL3 gene expression and the transabdominal testicular descent in rats. Methods On the 19th gestational day, the male fetuses that had been exposed to MBP on the 15th to 18th gestational days were dissected and the degree of testicular ascent was examined. Next, quantitative reverse transcriptase polymerase chain reaction was performed to analyze the testicular INSL3 gene expression. At 60 days of age, the testicular position was recorded in the rest of MBP-treated male offspring. Results On the 19th gestational day, INSL3 messenger RNA expression was significantly decreased in the MBP-treated testes, and the associated degree of testicular ascent was significantly higher than in the controls. At 60 days of age, 12 (54.5%) of 22 rats had either unilateral or bilateral UT. Conclusions Prenatal MBP may inhibit the INSL3 gene expression associated with the transabdominal testicular ascent in fetal rats, thereby causing UT in postnatal offspring.

Une atresie de l'oesophage avec double fistule tracheo oesophagienne (TEF) est une tres rare anomalie et le diagnostic preoperatoire de la TEF proximale est tres difficile. Ce papier decrit le cas d'une petite fille qui presentait une atresie de l'oesophage avec une double fistule. La fistule distale etait diagnostiquee avant l'operation alors que la fistule proximale etait decouverte pendant l'operation. Rater la presence d'une fistule proximale peut conduire a une morbidite augmentee et a une mortalite due aux infections respiratoires severes et a la necessite d'une seconde operation. Grand soin doit alors etre pris pour ne pas manquer la presence d'une TEF proximale chez les patients avec ce type d'anomalie.

It has been recently reported that valine, which was one of the branched chain amino acids, enhanced liver regeneration after a hepatectomy in rats. The aim of this study is to investigate the effect of enteral valine supplementation on the intestinal adaptation of short bowel syndrome using a rat model. Seven-week-old male Lewis rats underwent a 90% small bowel resection. The rats were randomly divided into two groups; Group V (valine-rich diet which contains valine, five times as the normal amount of valine as that found in standard rat chow) and Group S (standard rat chow), according to the diet each group received. The rats were killed and evaluated at the operative day, and postoperative days (POD) 7, 14, 30, and 60, respectively. The parameters of estimation were body weight (BW), a blood amino acids analysis, a urine organic acids analysis and a morphological examination of the residual small intestines. The BW and the intestinal wet weight, jejunal crypt depth and proliferating cell nuclear antigen positive cells in Group V at POD 7 were significantly higher than in Group S, while those in the Group V at POD 30 and 60 were smaller than in Group S. The urine methylmalonic acid (MMA) level in Group V at POD 30 and 60 was much higher than in Group S. The valine-rich diet was thus found to enhance intestinal regeneration after a small bowel resection in the acute phase. However, the long-term valine-rich diet supplementation was found to disturb the intestinal adaptation, which might be caused by the high production of MMA due to the valine-rich diet. This is the first report in which valine was used as a promoter of intestinal adaptation.

But: La disparition ou la diminution des cellules interstitielles de Cajal (ICCs) ont ete impliques dans de multiples desordres de la motilite intestinale humaine. Nous avons rencontre quelques cas souffrant d'une constipation severe ou d'enterocolite entrainant le deces des patients apres une operation definitive pour maladie de Hirschsprung, meme si des cellules intestinales ganglionnaires ont ete retrouvees. Nous explorons la distribution de ICCs en utilisant une coloration c-kit dans le segment normo ganglionnaire et comparons ces resultats avec le devenir clinique apres une operation definitive. Patients et Methodes: Les distributions de ICCs etaient explorees en utilisant une coloration c-kit dans le segment normo ganglionnaire dans l'intestin reseque de 15 cas de Maladie de Hirschsprung. La distribution de la 9,5 protein gene product (PGP 9,5) comme marqueur neuronal et ceux de NADPH-diaphorase (NADPHd) comme marqueur du neurone NO etait aussi examine. Le nombre de ICCs et de neurones etait evalue quantitativement. Les resultats histopathologiques etaient compares au devenir clinique apres l'operation definitive dans chaque cas. Resultats: Les cellules c-kit immunoreactives etaient de distribution normale dans le segment normo ganglionnaire dans 13 cas alors qu'ils etaient diminues de maniere majeure dans 2 cas (moins de 50%). Les repartitions de PCP 9,5 et de NADPHd etaient plus ou moins les memes dans tous les cas. Le transit intestinal des 13 cas montrant une distribution normale de c-kit etait satisfaisant. En contraste, le transit intestinal etait altere dans 2 cas avec un nombre diminue de cellules c-kit positives. Un enfant souffrait d'une constipation persistante severe et aussi avait subi une resection du colon dilate. L'autre enfant mourrait d'un sepsis en rapport avec une enterocolite post-operatoire secondaire a une dilatation majeure du colon. Conclusion: Une diminution marquee des cellules c-kit positives dans le segment normoganglionnaire peut nous permettre de predire un devenir difficile apres la chirurgie definitive, probablement en rapport avec une motilite intestinale de mauvaise qualite. De plus, examiner la distribution des cellules c-kit dans l'intestin reseque, chez les patients avec HD est considere comme obligatoire pour choisir le traitement post-operatoire dans chaque cas.

Summary Objective Persistent hyperinsulinaemic hypoglycaemia of infancy (PHHI) is a disorder of glucose metabolism that is characterized by dysregulated secretion of insulin from pancreatic β-cells. This disease has been reported to be associated with mutations of the sulfonylurea receptor SUR1 (ABCC8) or the inward-rectifying potassium channel Kir6.2 (KCNJ11), which are two subunits of the pancreatic β-cell ATP-sensitive potassium channel. Patients and Methods In 14 Japanese PHHI patients, all exons of SUR1 and Kir6.2 genes were analysed by polymerase chain reaction (PCR) and direct sequencing. Four patients responded to diazoxide, and nine patients underwent a subtotal pancreatectomy. Histologically, seven patients were diagnosed to have a focal form and two a diffuse form of the disease. Results We found nine novel mutations in the SUR1 gene and two in the Kir6.2 gene. In the SUR1 gene mutations, three were nonsense mutations (Y512X, Y1354X and G1469X), one was a one-base deletion in exon 7, and two were missense mutations in the nucleotide-binding domain 2 (K1385Q, R1487K). The other three mutations occurred in introns 14, 29 and 36, which might cause aberrant splicing of RNA. Two siblings in one family were heterozygotes for a missense mutation, K1385Q, which was maternally inherited. In Kir6.2 gene screening, one patient was found to be a compound heterozygote of a missense mutation (R34H) and a one-base deletion (C344fs/ter). Conclusion The novel mutations reported here could be pathological candidates for PHHI in Japan. They also reveal that SUR1 and Kir6.2 mutations in the Japanese population exhibit heterogeneity and that they occurred at a frequency similar to other genetic populations.

Background/Purpose The introduction of laparoscope and transanal endorectal pull-through has caused a revolution in the operative procedures for Hirschsprung's disease. To study the changing profile of Hirschsprung's disease in Japan, the authors carried out a national survey. Method Patient data were collected in 3 phases: group 1, 1628 patients between 1978 and 1982; group 2, 1121 patients between 1988 and 1992; and group 3, 1103 patients between 1998 and 2002, respectively. Results The incidence was 1:4697, 1:5544, and 1:5343 and the male/female ratio was 3.0:1, 3.4:1, and 3.0:1 in each group, respectively. Patients weighing less than 2500 g at birth increased to 10.4% in group 3, whereas they were 6.5% in group 2 and 5.5% in group 1. The patients with a family history also increased to 6.0% in group 3, in comparison with 2.8% in group 2 and 3.0% in group 1. The incidence of associated anomalies increased over time, 11.1% in group 1, 16.3% in group 2, and 21.2% in group 3. Mutations of genes were found in 4 of the 23 patients examined. The extent of aganglionosis was almost the same in each group. Regarding the definitive operation, the procedures without laparotomy, including transanal endorectal pull-through, increased up to 42.6% in group 3 but 0% in groups 1 and 2. The frequency that a primary operation without stoma was performed also increased. The age at definitive operation decreased in group 3. The incidence of preoperative and postoperative enterocolitis also decreased over time. The mortality was decreased over time, 7.1%, 4.9%, and 3.0% for groups 1, 2, and 3, respectively. Conclusions The authors analyzed 3852 Japanese patients over 3 decades. The ratio of patients with a low birth weight, associated anomalies, or a family history was increased in the last 10 years. A primary operation without laparotomy has thus become the procedure of choice for a definitive operation.

Objectives : Controversy remains regarding which residual segment, namely the jejunum or ileum, is more beneficial for intestinal adaptation in patients with short bowel syndrome (SBS). The aim of our study is to evaluate the residual intestinal adaptation after a massive small intestinal resection and thereby determine which of the two residual segments demonstrates a better intestinal adaptation. Methods : The SBS rats underwent about a 70 % resection of either the jejunum or ileum, respectively. Sham rats underwent an ileal transection with a subsequent reanastomosis of the same portion. The body weight was measured every day after the operation. Two weeks after the operation, all rats were sacrificed. The intestinal length, the hematological and serum chemical data, and the histological findings of the residual intestine were investigated in all rats to evaluate the intestinal adaptation. Results : The body weight gain in the rats with the residual ileum (ileum group) was similar to that of the Sham group and better than that of the rats with residual jejunum (jejunum group). The length of the residual intestines in the jejunum group was longer than that in both the sham and ileum groups. Regarding hematological and serum chemical studies, the jejunum group tended to demonstrate more anemia and malnutrition than both the sham and ileum groups. In a histological study, the villous height in both the jejunum and ileum groups was significantly larger than in the preoperative condition. In both groups, the degree of crypt depth only significantly increased in the ileum group in comparison to the preoperative condition. Regarding the thickness of the muscular layers, a no difference was observed among all groups. Conclusion : Based on our data, in patients with SBS, the use of the residual ileum was thereforefound to be preferable to that of the residual jejunum regarding intestinal adaptation.

Telomerase, an enzyme related with cellular immortality, has been extensively studied in many kinds of malignant tumours for clinical diagnostic or prognostic utilities. Telomerase activity is mainly regulated by the expression of hTERT (human telomerase reverse transcriptase), which is a catalytic component of human telomerase. To evaluate whether the levels of hTERT mRNA provides a molecular marker of hepatoblastoma malignancy, we examined hTERT mRNA expression levels in the primary hepatoblastoma tissues by fluorescent RT-PCR using LightCycler technology and followed up the clinical outcomes in 63 patients listed in the Japanese Study Group of Pediatric Liver Tumor between 1991 and 2002. The hTERT mRNA expression was detected in 61 (96.8%) specimens and their expression levels ranged between 0.1/1000 and 745.1/1000 copies of PBGD gene that was used as an internal control. Among these cases, frozen 39 tumour samples and 14 adjacent noncancerous liver tissues were analysed for semiquantitative telomerase assay. In the 39 tumour samples, the levels of telomerase activity ranged between 0.11 and 2709 TPG and 12 (30.7%) had high telomerase activity (>100 TPG), whereas only nine of 14 noncancerous liver tissue samples showed telomerase activity which was less than 1.0 TPG. The levels of telomerase activity were significantly correlated with the levels of hTERT mRNA expression (P

The survival outcome for patients with hepatoblastoma normally depends on the resectability of the tumor. In Japan, the pre and/or postoperative chemotherapy protocol using a combination of cisplatin (CDDP) and tetrahydropyranyl-Adriamycin (THP-ADR) has been the standard treatment since 1991. This study aims to assess exactly what influence the establishment of this chemotherapy protocol has had on both the tumor resectability and the outcome of patients with hepatoblastoma.From 1982 to 1997, 60 patients with hepatoblatoma were treated in the Kyushu area, Japan. Based on the pretreatment extent of disease (PRETEXT), the outcome and tumor resectability were compared between group A (1982 to 1990, n = 27, PRETEXT I:5, II:8, III:6, IV:8) and group B (1991 to 1997, n = 33, PRETEXT I:9, II:9, III:5, IV:10).The 5-year survival rates (group A and group B) were 33% and 73% for all cases (P.01), 100% and 89% for PRETEXT I, 38% and 89% for II (P.05), 17% and 80% for III (P.01), and 0% and 40% for IV (P.01), respectively. The 5-year survival rates for patients with metastases were 0% for group A (n = 5) and 57% for group B (n = 7; P.01). The rates of a complete resection of primary tumor were 48% for group A and 67% for group B. In particular, a significant difference was found regarding the complete resection rate between groups A and B in the patients with PRETEXT III (17% for group A and 80% for group B; P.01). In the patients with an incomplete tumor resection (14 for group A, 11 for group B), the 5-year survival rates were 0% for group A and 45% for group B (P.01).The optimal chemotherapeutic regimen of CDDP and THP-ADR was thus found to greatly contribute to the improved survival rate of hepatoblastoma patients. Preoperative chemotherapy resulted in an increased resectability of the tumor, whereas postoperative chemotherapy played an important role in the increased cure rate of cases with either an incomplete tumor resection or metastasis. However, refractory cases with PRETEXT IV or metastasis may still require the development of an even more effective treatment modality, including the use of blood stem cell transplantation.

Using light microscopic immunohistochemistry, neuron-specific enolase (NSE)-positive endocrine cells were quantitatively analyzed in the sheep lung during different stages of development from the canalicular stages to adulthood. In all stages, NSE-positive endocrine cells were usually located in the bronchi and bronchioles as solitary cells, although a few NSE-positive cell clusters, the so-called neuroepithelial bodies, were found in some places. The number of NSE-positive endocrine cells decreased with advanced stages of gestation. In the late alveolar stage, the number of NSE-positive endocrine cells reached its bottom during the fetal period. There was a gradual upturn after birth. The overall pattern of growth and differentiation of the endocrine cells is most likely species-related and depends on the state of airway development; the number of the endocrine cells of almost all animals, excluding the sheep, in relation to the size of the lung reaches a peak in the late fetal and early neonatal periods and decreases shortly thereafter. NSE-positive endocrine cells were also predominantly located in the large airways during the early stage of development (canalicular stage), and were found more frequently in the small peripheral airways towards the term. These results show the number of NSE-positive endocrine cells in the sheep to be different from that seen in other species.

We describe a 6-year-old girl and her mother with dominant β-thalassemia due to hemoglobin Hradec Kralove (Hb HK). Both patients presented microcytic anemia, jaundice, splenomegaly, cholelithiasis, and recurrent hemolytic bouts. Osmotic resistance tests using saline and coiled planet centrifugation revealed the increased fragility of the red cell membrane. On the other hand, the glycerol lysing time was prolonged, and results of the isopropanol test were weakly positive. Despite mimicking the features of hereditary spherocytosis, the results of the genetic analyses verified the second reported family with Hb HK (codon 115,\(G\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{C} C[Ala] \to G\underset{\raise0.3em\hbox{$\smash{\scriptscriptstyle-}$}}{A} C[Asp]\)). Splenectomy was effective for the amelioration of hemolysis. Of 7 reported patients with Hb variants at β-globin codon 115 (Hb Madrid and Hb HK), 5 underwent splenectomy. Because of the variable augmentation of extramedullary hemolysis in dominant β-thalassemias, genotyping is necessary for determining the clinical indication of splenectomy.

Mono-n-butyl phthalate (MBP) was administered to pregnant rats from the 15th to the 17th gestational day to investigate the dose-dependent effect of phthalate ester on testicular descent in both pre- and postnatal rats. Thirty pregnant rats (280-330 g) were separated into five groups and administered 1.0 g/kg/day MBP in group I, 0.5 g/kg/day in group II, 0.25 g/kg/day in group III, 0.125 g/kg/day in group IV from the day 15 to 17 of gestation. The rats in group V were only administered solvent and acted as controls. The position of the testis was evaluated in both pre- and postnatal rats. This was significantly higher in 20-day old fetuses in groups I and II than in groups III, IV or V. After birth, the incidence of undescended testis was also higher in groups I and II compared with those of groups III, IV or V. The undescended testes were located in the abdominal cavity in groups I and II, whereas they were located at the inguinal region in group III. The lowest dose that induced an adverse effect on testicular descent was 0.25 g/kg/day. In conclusion, maternal MBP inhibited testicular descent in prenatal rats and thus dose-dependently induced undescended testes postnatally. High-doses of MBP are thus considered to inhibit the transabdominal descent of the testis, probably due to its estrogenic activity, whereas low-doses of MBP may act as an anti-androgen and thereby inhibit inguinoscrotal testicular descent in postnatal rats.

We have managed four cases of Peutz-Jeghers syndrome (PJS) in children. Fathers of three of these patients had PJS. There was also a family history of cancer in three cases, with pancreatic cancer in a father, colonic and laryngeal cancers in a grandfather, and hepatic and gastric cancers in a grandmother. It is presumed that in each of the cases, the largest polyp was responsible for initial symptoms. Preoperative examination revealed additional small polyps in the whole alimentary tract except for the oesophagus. Patients underwent laparotomy to remove the largest polyps and subsequent intraoperative endoscopic polypectomy for other small polyps, to minimize intestinal resection. Follow-up gastrointestinal examinations, including upper gastrointestinal series, small intestinal contrast study, and barium enema, were repeated about once a year. Three of four cases showed recurrent small intestinal polyps, and one required a second laparotomy because of recurrent abdominal pain. In conclusion, patients with PJS occurring in childhood have a strong hereditary family history of cancer and a high incidence of recurrence. Careful follow-up examination is mandatory for the gastrointestinal tract, as well as other solid organs that are susceptible to malignant change, throughout a patient's life.

Morphometric analyses of the immunohistochemical expression of surfactant apoprotein A (SP-A) were carried out on the bronchioles of human congenital diaphragmatic hernia (CDH) neonates and then compared with those in a gestational and postnatal age-matched control group. There was no difference in SP-A expression between lung specimens of the control group and unaffected lungs in the CDH group. However, compared with both these lungs, the ipsilateral lungs of the CDH group showed a significant increase in SP-A expression, namely, the number of SP-A-positive cells per bronchiole, the number of SP-A-positive cells per unit perimeter of bronchiole, and the number of SP-A-positive cells per unit bronchiolar surface area. These results suggest that in lungs of CDH cases, especially ipsilateral lungs, there is a possible delay in the functional maturation or development of SP-A synthesis by the bronchiole, and this retardation may play a role in the postnatal respiratory insufficiency observed in CDH patients.

Interstitial cells of Cajal (ICCs) have been reported to play the role of a pacemaker in regulating bowel motility. The relationship between neurons and ICCs, however, remains unclear. Hirschsprung's disease (HD) is an ideal model for investigating this relationship. The operated specimens obtained from 6 short and 3 long segment aganglionosis patients and 3 controls were used as the subject materials in this study. ICCs were immunohistochemically identified using a specific antiserum c-kit, a tyrosine kinase receptor expressing ICCs. Nitrergic nerves were demonstrated by NADPH-diaphorase (NADPH-d) histochemistry. C-kit immunohistochemistry was also combined with protein gene product 9.5 (PGP 9.5; as a general neuronal marker). In the normoganglionic segment of HD, numerous c-kit-positive cells and NADPH-d positive neurons were found in the proper muscle layer, including Auerbach's plexus. In the oligoganglionic segment, the number of c-kit-positive cells and NADPH-d neurons slightly decreased. In the inner border of the circular muscle layer (IBCM), the c-kit-positive cell networks and NADPH-d activities remained in short segment cases, while both of them were absent in the long segment cases. In the aganglionic segment, c-kit positive cells were present universally but the number of them was slightly decreased in the proper muscle layer. The c-kit-positive cell networks of IBCM were seen where extrinsic neurons were present, while they were almost completely absent where extrinsic neurons were absent in the proximal zone of the long segment cases. C-kit positive cells were present universally in the oligoganglionic as well as aganglionic segments of HD. The distribution and properties of c-kit positive cells were related to the presence of extrinsic neurons in aganglionic segment. Based on these findings, aperistalsis is considered not to relate with c-kit positive cells, and c-kit positive cells are not supposed to have a neurogenic origin and can develop without neurons, however the lack of enteric neurons may influence the full differentiation of ICCs.

Purpose: Allogenic blood transfusions have a risk of infection owing to unknown organisms, graft-versus-host reaction, and immunosupression; however, the use of autologous blood has been reported to be safe. Cord blood has been reported to be useful as a source of stem cell transplantation for the treatment of leukemia and genetic disease. Furthermore, autologous cord-blood transfusions (ACBT) have been reported to be effective for the treatment of anemia in premature infants. The authors examined the efficacy of ACBT in neonatal surgical patients. Methods: Autologous cord-blood was stored from 12 infants at delivery, including 2 transvaginal and 10 cesarean section deliveries, from 1998 to 2001. All infants had surgically correctable malformations diagnosed antenatally. The mean gestational age was 37.2 ± 1.6 weeks, and the birth weight was 2,597 ± 1.6 g. The results of the blood count, serum electrolyte, and liver function tests of the patients who underwent ACBT only (group 1, n=7) were compared with those of the 7 neonates who underwent an allogenic transfusion during the same period (group 2, n=7). Results: The mean volume of the stored blood was 64 ± 35.6 g (range, 20 to 100). Eleven of the 12 patients underwent transfusions. Ten of 11 patients received autologous cord blood. A mean of 44.1 ± 37.3 g of cord blood was used. Three of 10 cases also required an allotransfusion because of ECMO circuit preparation and a shortage of the stored blood. One patient underwent allotransfusion only. As a result, 7 of 11 babies (64%) who required transfusion were able to avoid an allotransfusion. The blood potassium levels were lower in group 1 than in group 2. No significant complications were recognized clinically. Conclusions: ACBT is considered beneficial because it enables neonatal surgical patients to avoid allotransfusions. Therefore, autologous cord-blood storage should be considered in the patients antenatally diagnosed to have surgical malformations. However, the storage volume varies for each case. Improved techniques to obtain an adequate amount of blood also should be developed. J Pediatr Surg 38:604-607. Copyright 2003, Elsevier Science (USA). All rights reserved.

Background/Purpose: Intestinal dysmotility is an important problem in the postoperative management of patients with intestinal atresia (IA). Changes in the intramural components have so far been histochemically and immunohistochemically examined in both the proximal and distal segments of IA, but no detailed analysis of the muscular elements has been performed. The aim of this study was to carefully examine any alterations in the muscular elements in the intestines from patients with IA. Methods: Resected intestines were obtained from 6 patients with ileal atresia, 4 patients with jejunal atresia, and 3 controls without gastrointestinal diseases obtained by autopsy (congenital diaphragmatic hernia). All specimens were immunochemically stained with a monoclonal antibody to α-smooth muscle actin (α-SMA) as a smooth muscle marker. Results: In the normal small intestine, almost all the enteric smooth musculature were positive for α-SMA antiserum, except for the bulk of the circular musculature. In the proximal segments of all cases with IA, a reduced staining intensity for α-SMA was observed mainly in the severely hypertrophic muscle layers. In addition, some bundles of smooth muscle also were located in the submucosal connective tissue near the border of the innermost layer of the circular musculature, in which large amounts of smooth muscle fibers extended occasionally from the innermost layer of the circular musculature to the muscularis mucosae in the proximal segments of 4 cases. In the distal segments of IA, the distribution of α-SMA–positive smooth muscle fibers was similar to that in the control intestines, excluding mild to moderate hypertrophy of the muscular layers. Conclusions: Both severe hypertrophy and a reduced immunoreactivity for α-SMA were observed in the circular muscle layer of the proximal segments. In addition, the occurrence of α-SMA–positive abnormal smooth muscle fibers was recognized in the submucosal layers of the proximal segments, thus, suggesting a delay in the intestinal muscular formation or a regressive reaction secondary to dilatation. These muscular alterations in the proximal segments might be considered to contribute to the postoperative intestinal dysmotility in IA cases. J Pediatr Surg 38:161-166. Copyright 2003, Elsevier Science (USA). All rights reserved.

Patients with high-risk neuroblastoma who have multiple copies of MYCN fare much worse than do those without MYCN amplification; however, it has not been clarified whether intensified chemotherapy with or without blood stem cell transplantation can alter the extremely poor prognosis of patients with amplified MYCN.Between 1985 and 1999, 301 patients older than age 12 months with stage 4 neuroblastoma were treated. From January 1985 to February 1991, 80 patients with stage 4 neuroblastoma with and without MYCN amplification uniformly received induction chemotherapy with regimen A(1) (cyclophosphamide 1,200 mg/m(2) and vincristine 1.5 mg/m(2) on day 1, tetra-hydropyranyl [THP]-Adriamycin 40 mg/m(2) on day 3, and cisplatin 90 mg/m(2) on day 5). Among 22 patients with MYCN amplification, nine (40.9%) achieved a complete remission and seven (31.8%) underwent stem cell transplantation. Of 58 patients without MYCN amplification, 43 (74.1%) achieved a complete remission and 14 (24.1%) underwent stem cell transplantation. The 5-year relapse-free survival rates were 23.2% for stage 4 patients with MYCN amplification and 33.3% for those without MYCN amplification (P = 0.029); the 5-year overall survival rates were 32.8% for stage 4 patients with MYCN amplification and 42.8% for those without MYCN amplification (P0.05). From March 1991 to June 1998, patients with stage 4 neuroblastoma who had 10 or more copies of MYCN were treated with regimen A(3) (cyclophosphamide 1,200 mg/m(2) per day on days 1 and 2, THP-Adriamycin 40 mg/m(2) on day 3, etoposide 100 mg/m(2) per day on days 1 to 5, and cisplatin 25 mg/m(2) per day on days 1 to 5); those with fewer than 10 copies of MYCN received regimen new A (cyclophosphamide 1,200 mg/m on day 1, THP-Adriamycin 40 mg/m on day 3, etoposide 100 mg/m per day on days 1 to 5, and cisplatin 90 mg/m on day 5), which is similar in intensity to regimen A. Among 88 patients with MYCN amplification, 63 (71.6%) achieved a complete remission and 63 (71.68%) underwent stem cell transplantation. Of 133 patients without MYCN amplification, 93 (69.9%) achieved a complete remission and 71 (53.4%) underwent stem cell transplantation. The 5-year relapse-free survival rates were 36.0% for stage 4 patients with MYCN amplification and 32.2% for those without MYCN amplification (P0.05), the 5-year overall survival rates were 34.0% for stage 4 patients with MYCN amplification and 38.9% for those without MYCN amplification (P0.05). The difference in relapse-free survival rates was significantly different (P = 0.003) between patients with MYCN-amplified tumor treated before (regimen A(1)) versus after 1991 (regimen A(3)).With the use of the more intensive induction regimen A plus blood stem cell transplantation for MYCN-amplified patients, survival curves for those with or without MYCN amplification now appear similar. Higher doses of chemotherapy may ameliorate the effect of MYCN amplification in patients with high-risk neuroblastoma.

Background/Purpose: Since 1985, a nationwide mass screening program (MS) for neuroblastoma has been conducted for 6-month-old infants throughout Japan, resulting in the detection of more than 1,900 cases of neuroblastoma. The outcome of these patients has been excellent: more than 97% of them are alive. Yet, several reports suggest that the number of advanced-stage neuroblastoma patients over 1 year of age has not changed substantially. The current report focuses on the 15-year experience with MS of the Kyushu Pediatric Oncology Study Group. Methods: The clinical and biological features of neuroblastoms detected (n = 320) and not detected by MS (n = 245) were compared. Regional and national statistics for neuroblastoma before and after 1985 were analyzed using standard epidemiologic measures for the occurrence of disease. Results: The majority of the MS-positive cases were biologically favorable and had an excellent outcome. In contrast, the majority of non-MS patients in whom neuroblastoma later developed had advanced-stage, unfavorable-prognosis tumors. The overall mortality rate of neuroblastoma in the Kyushu area was not improved by MS. Conclusions: The optimal time for screening is the point at which neuroblastomas regressing spontaneously can no longer be detected, but more aggressive disease can be found. A birth cohort study could determine the optimal timing for a second screening. Identification of other new prognostic factors may be required. J Pediatr Surg 37:949-954. Copyright 2002, Elsevier Science (USA). All rights reserved.

The International Neuroblastoma Staging System and Pathology Classification were proposed in 1988 and in 1999, respectively, but their clinical value has not yet been fully studied in new patients. Six hundred and forty-four patients with neuroblastoma treated between January 1995 and December 1999 were analysed by these classifications. The 4-year overall survival rate of patients

Objectives: We investigated the effect of nucleosides (NSs) and nucleotides (NTs) on the intestine and intestinal graft in a model of syngenic small bowel transplantation, with the fetal rat intestine as a graft. Methods: Two-centimeter jejunal segments from Lewis rats at 19 d of gestation were transplanted into the abdominal walls of 5-wk-old Lewis rats by using a non-vascular anastomotic technique. After transplantation, the rats were assigned to one of two groups: group 1 did not receive NS or NT and group 2 was supplemented with NS and NT. The grafts and graft recipients were examined morphologically 14 d after transplantation according to conventional histologic and immunohistochemical studies of neurons and smooth muscles. Results: Group 1 gained little body weight, even though both groups received similar amounts of food. The grafts in group 1 showed poor development in length, diameter, and wet weight. They also showed poor villi development, abnormalities in nerve distribution, and degeneration of muscle layer structure on histologic and immunohistochemical studies. Conclusions: We found that NS and NT are essential nutrients for intestinal growth and maintenance of structures in fetal small bowel transplantation.

Purpose: The authors used a magnetic spinal stimulation technique to assess both the preoperative sphincter function and the effect of operative intervention on the sphincter muscle in patients with anorectal malformations. Methods: The authors examined 5 patients with an anorectal malformation consisting of 4 high-type cases and 1 intermediate-type case, that all had undergone posterior sagittal anorectoplasty during the previous 2-year period. Control data also were obtained from 11 age-matched healthy children. A contraction of the external anal sphincter (EAS) was evoked by magnetic stimulation of the sacral roots with a figure-of-eight coil at the level of the lumber and sacral segments. EAS electromyograms then were recorded at the anal verge. Results: The preoperative motor latencies at L4 stimulation increased significantly more in the patients than in the controls (5.9 ± 0.5 msec v 4.1 ± 0.7 msec during left side recording). In the postoperative examination (1 to 5 months after operation), the mean maximum amplitude decreased slightly without any significant difference, whereas the latency was almost identical to the preoperative data. Conclusion: These results suggest that the neurologic function of EAS was congenitally disturbed, and the current operative technique had no significant effect on the sphincter function. J Pediatr Surg 37:617-622. Copyright 2002, Elsevier Science (USA). All rights reserved.

Purpose: To determine the dose-limiting toxicity, maximum tolerated dose, and potential efficacy of irinotecan in children with refractory malignant solid tumors. Patients and Methods: In the present phase I clinical trial, 28 patients received irinotecan 50 to 200 mg/m 2 per day by intravenous 2-hour infusion over the course of 3 days, repeated once after an interval of 25 days. Fifty-one treatment courses were administered to these patients. Results: Dose-limiting toxicities were observed at the dose of 200 mg/m 2 per day for 3 days. Diarrhea and hematopoietic toxicities were the dose-limiting factors, and the former required support with intravenous fluid administration. The occurrence of vomiting was variable. Decreases in clinical tumor marker levels were observed in the majority of patients who received two cycles of irinotecan 80 mg/m 2 per day to 200 mg/m 2 per day over the course of 3 days, and partial response was attained in four patients who received irinotecan in two cycles of 140 mg/m 2 per day to 200 mg/m 2 per day over the course of 3 days. Pharmacokinetic studies showed that the plasma concentration of irinotecan and its active metabolite SN-38 ranged from 93 to 2,820 ng/mL and 5.2 to 34.8 ng/mL, respectively, during 3-day infusions of irinotecan 200 mg/m 2 per day. The mean clearance of irinotecan was 14.54 L/h per m 2 (range 8.45-20.83 L/h per m 2 ). Conclusion: The maximum tolerated dose was determined to be a dose of irinotecan between 160 mg/m 2 per day and 180 mg/m 2 per day administered over the course of 3 consecutive days on an inpatient basis, repeated once after 25 days off, and our results indicate that irinotecan is a promising anticancer agent that is worthy of phase II trials in pediatric solid tumors.

We report a case with vesicoureteral reflux (VUR), diagnosed antenatally. A 31-year-old Japanese woman was referred at 37 weeks’ gestation because of the fetus having unilateral renal pelvis dilatation. The real-time ultrasound revealed a right renal pyelectasia with periodic changes in size over intervals lasting several minutes. The simultaneous examinations of both anteroposterior dimension of the renal pelvis and the ellipse size of urine-filled bladder area demonstrated that they varied in size in a reciprocal fashion, leading us to diagnose the fetus as having VUR. Postnatally, bilateral VUR was confirmed by the voiding cystogram. This case has typical ultrasonographic findings of fetal VUR, in which both the renal pelvis and urinary bladder dimensions change periodically in a reciprocal manner with each other, when the reflux really occurred.

We report a case of congenital pyloric atresia in which two cystic masses in the fetal upper abdomen and polyhydramnios were observed from 24 weeks’ gestation to term. The two cystic masses were thought to indicate the ‘double bubble’ sign in utero, leading to a misdiagnosis of congenital duodenal obstruction. At 33 weeks’ gestation, continuous ultrasound observation of the two cystic masses was made for 60 min. During the periods when gastric peristalsis was absent, the ‘double bubble’ sign was observed. However, during the periods when gastric peristalsis was present, the ‘double bubble’ sign disappeared. In retrospect, both cystic masses were stomach and the ‘double bubble’ sign disappeared with peristalsis of the antrum. In order to differentiate from congenital duodenal obstruction, whole stomach configuration should be delineated by continuous observation covering periods when gastric peristalsis is active as well as quiet.

Background and Methods. The prognosis in neuroblastoma, which is the most common solid tumor in children, tends to vary greatly, and many studies have demonstrated both clinical and biological factors to be closely correlated with the outcome. In order to select the optimal treatment according to the degree of malignancy of neuroblastoma, it is essential to accurately and rapidly identify any genetic heterogeneity associated with the prognosis. We assessed the status of some genetic abnormalities (MYCN amplification, deletion of the short arm of chromosome 1, DNA ploidy, and a gain of the chromosome 17q region) associated with the prognosis using several moleculat biological methods. Results and Conclusions. The combination of several molecular biological techniques is thus considered to be useful for elucidating the degree of malignancy of neuroblastoma. In particular, diagnostic analyses based on a combination of the fluorescence in situ hybridization (FISH) method and the quantitative polymerase chain reaction (PCR) method may be considered to be the most effective methods for quickly and accurately evaluating any aberrations in the gene dosages associated with the patients' outcomes. (Surgery 2002;131:S283-7.)

Background. In Hirschsprung's disease, the severity of bowel obstruction varies among those patients who have the affected colon of a similar length, suggesting that there is more than a simple aperistaltic obstruction in the pathophysiology of Hirschsprung's disease. Methods. A series of our electrophysiological studies of the aganglionic segments from human specimens and rat models were reviewed to obtain an overview of Hirschsprung's disease. Results. In human studies, a generation of regular spontaneous activity was recorded in both the dilated ganglionic segment and transitional aganglionic region, while the smooth muscle cells of the narrow aganglionic segment were electrically quiescent. According to a pattern of innervation, in the dilated ganglionic segment inhibitory junction potentials associated with or without excitatory junction potentials were observed in all of the examined cells, and these intrinsic nervous inputs were gradually decreased in the transitional region. In the narrow aganglionic segment, only excitatory junction potentials of the extrinsic nervous origin were found in about 20% of the examined cells In rat models, distally increasing tendency of the excitatory nervous inputs was observed in the narrow aganglionic segment. Conclusion. A bowel obstruction in Hirschsprung's disease might be generated due to complex mechanisms involving myogenic and neurogenic abnormalities. (Surgery 2002;131:S288-93.)

Objective To investigate whether prenatal maternal stress, considered to alter plasma testosterone concentration and induce a lack of testosterone surge in male rat fetuses, has any effect on the growth of the processus vaginalis and testicular descent in male offspring. Materials and methods Pregnant rats were divided into two groups. In group 1, the rats were placed three times daily for 60 min each session in Plexiglas rat holders (13×6×8 cm) illuminated by two 150-W flood lights from day 14 to day 18 of gestation during the dark phase of the animals’ light (12 h) and dark (12 h) cycle. In group 2, pregnant females were not handled and acted as controls. After birth, the anogenital distance of both the male and female rats was measured, and the length of the processus vaginalis was measured at 7 days of age in some rats. Thereafter, at 21, 24, 27 and 30 days of age, testicular descent was assessed in the remaining male rats. Results In group 1, at 21, 24, 27 days of age, testicular descent was delayed in the stressed rats compared with the control rats. At 30 days of age, five of 36 stressed male offspring showed abnormal testicular descent; three testes were in the superficial inguinal position and two were above the scrotum. Both the anogenital distance and the length of the processus vaginalis were significantly less in the stressed male rats than in the controls. Conclusions Maternal stress induced a female-like anogenital distance in male rat fetuses, and caused delayed and abnormal descent of the testis, by inhibiting the growth of the processus vaginalis after birth. Prenatal stress may therefore be involved in the delay of testicular descent in neonatal rats by inducing a mild anti-androgenic effect in the fetus.

The mechanism underlying the LTC4-induced contraction of guinea-pig taenia coli was determined using the simultaneous measurements of [Ca2+]i and force in whole muscle preparations. Additional experiments were performed in receptor coupled permeabilized preparation. For comparison purposes, the contraction which was induced by a typical G-protein mediated agonist, carbachol was also characterized. LTC4 induced a contraction in the guinea-pig taenia coli in a concentration-dependent manner. The maximal response was obtained at 100 nM and the EC50 value was 5.4±1.9 nM. Both LTC4 and carbachol induced increases in [Ca2+]i and force. The maximum force induced by 100 nM LTC4 was significantly smaller than that induced by 10 μM carbachol, although an increase in [Ca2+]i produced by both agonists was similar. In the permeabilized preparations, carbachol, but not LTC4, induced an additional force development at a fixed Ca2+ concentration. LTC4 induced no increase in [Ca2+]i and force in the Ca2+-free solution, while carbachol induced transient increases in both [Ca2+]i and force in a Ca2+-free solution. Both diltiazem and SK&F 96365 significantly inhibited the LTC4− and carbachol-induced increases in [Ca2+]i and force in normal PSS. The inhibitory pattern of [Ca2+]i by these drugs was also similar. We thus conclude that LTC4 induces the contraction of the guinea-pig taenia coli mainly through Ca2+ influx via both the diltiazem-sensitive and SK&F 96365-sensitive Ca2+ channels, without affecting either the Ca2+-sensitivity or the intracellular Ca2+ release. These results indicated that the mechanism underlying the LTC4-induced contraction differs greatly from that for conventional G-protein mediated agonists, such as carbachol. British Journal of Pharmacology (2001) 133, 529–538; doi:10.1038/sj.bjp.0704122

Malignant rhabdoid tumor (MRT) is characterized by the presence of intracytoplasmic eosinophilic inclusions composed of whorls of intermediate filaments. This tumor was originally described as an entity of the abortive type of Wilms' tumor in childhood. Recently, it has been proved that these rhabdoid cells can be observed in various types of malignant tumors, including soft tissue sarcoma or carcinoma. To investigate the oncogenesis of this tumor, we examined the p53 gene alteration by means of immunohistochemical analysis and DNA direct sequencing in three cases of malignant rhabdoid tumor (MRT) of the soft tissue and three cases of MRT of the kidney. All the cases of MRT of the soft tissue and two of the cases of MRT of the kidney showed immunopositivity for p53 protein. Among them, one of the cases of MRT of the soft tissue and two of the cases of MRT of the kidney showed missense mutations of the p53 gene. These results strongly suggest that p53 gene alterations may have an important role to play in the aggressive biological behavior and poor prognosis of this tumor.

The amplification of the N-myc gene and a gain of the chromosome 17q arm correlate with an unfavorable outcome in patients with neuroblastoma. In this study, we determined the gene dosage of the N-myc gene (located at 2p24) and Survivin gene (located at 17q25) using the p53 gene (located at 17p13) as the internal control gene by the TaqMan polymerase chain reaction (PCR)-based gene dosage analysis in 25 neuroblastoma samples. Based on the assumption that the gene dosages of each gene of a normal individual lymphocytes are 1.0, 11 of the 25 cases with a corrected gene dosage of N-myc (N-myc/p53) of more than 2.0 had a more unfavorable prognosis than the 14 cases with a N-myc gene dosage of less than 2.0 (5-year survival rate: 18 vs. 71%, P