池田 太郎

© 2020, The Author(s). Purpose: Our previous studies demonstrated that mature adipocyte-derived dedifferentiated fat (DFAT) cells possess similar multipotency as mesenchymal stem cells. Here, we examined the immunoregulatory potential of DFAT cells in vitro and the therapeutic effect of DFAT cell transplantation in a mouse inflammatory bowel disease (IBD) model. Methods: The effect of DFAT cell co-culture on T cell proliferation and expression of immunosuppression-related genes in DFAT cells were evaluated. To create IBD, CD4+CD45RBhigh T cells were intraperitoneally injected into SCID mice. One week later, DFAT cells (1 × 105, DFAT group) or saline (Control group) were intraperitoneally injected. Subsequently bodyweight was measured every week and IBD clinical and histological scores were evaluated at 5 weeks after T cell administration. Results: The T cell proliferation was inhibited by co-cultured DFAT cells in a cell density-dependent manner. Gene expression of TRAIL, IDO1, and NOS2 in DFAT cells was upregulated by TNFα stimulation. DFAT group improved IBD-associated weight loss, IBD clinical and histological scores compared to Control group. Conclusion: DFAT cells possess immunoregulatory potential and the cell transplantation promoted recovery from colon damage and improved clinical symptoms in the IBD model. DFAT cells could play an important role in the treatment of IBD.

PURPOSE: Our previous studies demonstrated that mature adipocyte-derived dedifferentiated fat (DFAT) cells possess similar multipotency as mesenchymal stem cells. Here, we examined the immunoregulatory potential of DFAT cells in vitro and the therapeutic effect of DFAT cell transplantation in a mouse inflammatory bowel disease (IBD) model. METHODS: The effect of DFAT cell co-culture on T cell proliferation and expression of immunosuppression-related genes in DFAT cells were evaluated. To create IBD, CD4+CD45RBhigh T cells were intraperitoneally injected into SCID mice. One week later, DFAT cells (1 × 105, DFAT group) or saline (Control group) were intraperitoneally injected. Subsequently bodyweight was measured every week and IBD clinical and histological scores were evaluated at 5 weeks after T cell administration. RESULTS: The T cell proliferation was inhibited by co-cultured DFAT cells in a cell density-dependent manner. Gene expression of TRAIL, IDO1, and NOS2 in DFAT cells was upregulated by TNFα stimulation. DFAT group improved IBD-associated weight loss, IBD clinical and histological scores compared to Control group. CONCLUSION: DFAT cells possess immunoregulatory potential and the cell transplantation promoted recovery from colon damage and improved clinical symptoms in the IBD model. DFAT cells could play an important role in the treatment of IBD.

Objective: In recent years, improved survival rates of extremely low birth weight infants (ELBWIs) have led to an increasing number of enterostomy performed for those with meconium obstruction of prematurity (MOP)(1,2), spontaneous intestinal perforation (SIP)(3,4). To prevent serious stoma-related complications such as stoma side perforation, prolapse, fall and surgical site infection, we introduce our new "sutureless enterostomy" technique. Methods: We present the procedures in detail. We reviewed the medical records of twelve patients who underwent "sutureless enterostomy" in our neonatal intensive care unit from 2007 to 2013. Patient attributes, surgery-related items, stoma-related complications and outcomes were investigated. Results: Mean birth weight was 671 +/- 158 g (mean +/- S.D.). Six cases of MOP, three cases of SIP and three cases of NEC were diagnosed. Mean operative time was 75 +/- 35 min (mean +/- S.D.) None of them presented any of early stoma-related complications (necrosis, fall, and surgical site infection). However the parastomal hernia occurred in one patient as late complication. Three deaths occurred postoperatively as a result of exacerbations of their conditions. Conclusions: Based on our preliminary observations, our new "sutureless enterostomy" was done safely and reduced the risk of stoma-related complications. It may be an ideal procedure for the ELBWI with MOP or SIP. (C) 2017 Elsevier Inc. All rights reserved.

6歳女児。間欠的腹痛と非胆汁性嘔吐を主訴とした。5歳時に卵管留水腫を伴う卵管捻転に対し捻転解除および卵管開口術を施行し、外来フォロー中であった。腹部超音波検査にて腸間膜リンパ節の軽度腫大を認め、骨盤CTでは骨盤内直腸腹側に径4cm大のlow density massを認めたが、明らかな造影効果は認めなかった。右卵管捻転の再発を疑いで緊急手術を施行し、右卵管は卵管膨大部で反時計方向に360度の捻転を起こしていたが、卵巣には異常を認めなかった。卵管捻転の原因となる器質的異常は認めず、捻転解除後、卵巣を子宮後壁に吸収糸を用いて固定した。術後経過は良好で第3病日に退院し、固定術後3年経過して再発は認めていない。

The management of patients with acute perforated appendicitis with abscess is controversial. The aim of the present study was to assess the outcomes of treatment in patients with this condition. We retrospectively analyzed 31 patients (16 men and 15 women with a mean age of 8.4 years) with appendicitis presenting with abscess. Patients were divided into two groups (emergency operation group and interval operation group), and clinical characteristics and outcomes of treatment were investigated. On presentation, no differences in gender, age, body weight, duration of symptoms, temperature, white blood cell count, C-reactive protein level, or maximum size of the abscess in the axial view were detected between the two groups. Fifteen patients (48.4 %) underwent emergency surgery. The remaining 16 patients (51.6 %) were initially treated conservatively with antibiotics. All 16 patients underwent planned operations after receiving conservative treatment, and two (12.5 %) of these patients underwent appendectomy before the planned operation day because of recurrent appendicitis without abscess. There were no differences in the length of hospital stay. In the emergency operation group, six (40 %) patients presented with wound infection and four (26.7 %) developed a postoperative intra-abdominal abscess. No infective complications were reported in the interval operation group. Interval appendectomy after conservative treatment of pediatric ruptured appendicitis with abscess significantly reduced postoperative infection rates.

Background/purpose: Mainstreammodels for anal sphincter injury use large animals. We developed a simple and stable anal sphincter injury model in a small animal (i.e., rats) to obtain manometry measurements by using a miniaturized probe and applying cardiotoxin. Methods: The histological structure of the anal canal was evaluated by using manometry in normal rats (n = 40). We damaged the internal and external anal sphincters by locally administering snake poison (cardiotoxin; 20 uM, 100 mu L 8 points). We evaluated the anal canal function through manometry measurements (n = 5) and examined the histology using hematoxylin-eosin staining (at each time point, n = 3; total n = 15). Results: The manometry parameters and structure of the anal canal of normal rats were similar to those of humans, because rats have resting pressure, rectoanal reflex in the manometry, and an external and internal anal sphincter. After inducing injury, the following findings were observed: rhythmic wave loss and a remarkable reduction in the anal sphincter resting pressure; and local bleeding and advanced infiltration of the inflammatory cells (day 1) and the loss of muscle fibers (day 3). Conclusion: This new rat model will contribute to increasing the knowledge on the anal canal. (C) 2015 Elsevier Inc. All rights reserved.

The current study aimed to verify the usefulness of preoperative ultrasonographic evaluation of contralateral patent processus vaginalis (PPV) at the level of the internal inguinal ring. This was a prospective study of patients undergoing unilateral inguinal hernia repair at two institutions during 2010-2011. The sex, age at initial operation, birth weight, initial operation side, and the preoperative diameter of the contralateral PPV as determined using ultrasonography (US) were recorded. We analyzed the incidence of contralateral inguinal hernia, risk factors, and the usefulness of the preoperative major diameter of the contralateral PPV. The follow-up period was 36 months. All 105 patients who underwent unilateral hernia repair completed 36 months of follow-up, during which 11 patients (10.5 %) developed a contralateral hernia. The following covariates were not associated with contralateral hernia development: sex (p = 0.350), age (p = 0.185), birth weight (p = 0.939), and initial operation side (p = 0.350). The preoperative major diameter of the contralateral PPV determined using US was significantly wider among patients with a contralateral hernia than those without a contralateral hernia (p = 0.001). When the 105 patients were divided into two groups according to cut-off values of the preoperative major diameter of the contralateral PPV (wide group, > 2.0 mm; narrow group, a parts per thousand currency sign2.0 mm), a significant association was observed between the preoperative major diameter of the contralateral PPV and patient outcomes (p = 0.001). We used US and confirmed the usefulness of a preoperative evaluation of the major diameter of the contralateral PPV at the level of the internal inguinal ring in pediatric patients with unilateral inguinal hernias.

48歳男。出生時に高位鎖肛の診断で会陰式肛門形成、月齢3に直腸尿道瘻の診断で人工肛門造設術、2歳時に腹会陰式直腸肛門形成術、3歳時に人工肛門閉鎖術、15歳時に術後粘膜脱に対する形成術を受けていた。排便機能は不良で、数年毎に外来通院しており、45歳時より直腸粘膜脱や便失禁のため通院回数が増加し、48歳時に排便困難や残便感、排尿時違和感のため来院した。骨盤造影CTで直腸と前立腺の間に不整形腫瘤を認め、大腸内視鏡では下部直腸に易出血等の粘膜不整を認めた。確定診断のため人工肛門造設と直腸病変に対する生検術を施行し、病理組織所見より直腸癌と診断して直腸切断術を施行した。術中所見で前立腺への浸潤が疑われたため、前立腺の一部を合併切除した。病理診断はRb/P、Type 4(40×40mm)、poorly differentiated tubular adenocarcinoma and mucinous adenocarcinoma、INFc、ly3、v3、pAI(prostate)、scirrhous type、pN3で、stage IIIbであった。術後化学療法を施行したが、多発肝転移と肝不全で術後1年4ヵ月に死亡した。

目的:腹膜転移による大腸癌再発に認められた悪性小腸イレウスの臨床的特徴を分析した。方法:腹膜転移による大腸癌再発で悪性小腸イレウスを呈した7例(男性5例、女性2例、52〜78歳、平均66.4歳)について、その臨床的特徴を分析した。成績:原発は下行結腸2例(28.6%)、S状結腸1例(14.3%)、直腸4例(57.1%)で、直腸に多く認めた。全例がP2以上(P2;1例、P3;6例)の腹膜転移、H1以上(H1;1例、H2;5例、H3;1例)の肝転移、N2以上(N2;1例、N3;6例)のリンパ節転移を認め、腹腔以外の遠隔転移は肺転移を3例(42.9%)に認めた。すなわち、全例が高度進行再発癌であった。組織型は分化型(tub2;3例)が3例(42.9%)、未分化型(por;1例、muc;3例)が4例(57.1%)で、未分化型を多く認めた。小腸の閉塞・狭窄部は、全例が空腸から回腸までの3ヶ所以上に認めた。手術は胃瘻2例(28.6%)、腎瘻1例(14.3%)、胃瘻+腎瘻1例(14.3%)、試験開腹3例(42.9%)であった。再手術から死亡までの期間は0.5〜1ヵ月3例(42.9%)、1〜1.5ヵ月3例(42.9%)、1.5〜3ヵ月1例(14.3%)で、全例が3ヵ月以内に死亡した。結論:腹膜転移による大腸癌再発症例で、悪性小腸イレウスを呈するものは極めて予後不良であった。(著者抄録)

To clarify the physiologic function of the enteric nervous system (ENS) in the elongated sigmoid colon (ESC) of patients with sigmoid volvulus (SV), we examined the enteric nerve responses in lesional and normal longitudinal muscle strips (LMS) derived from patients with ESC and patients who underwent colon resection for colonic cancers. Thirty preparations of LMS were taken from the lesional sigmoid colons of 10 ESC patients with SV (8 men and 2 women, aged 53 to 80 years, mean 66.2 years). Forty preparations of LMS were taken from the normal sigmoid colons (NSC) of 20 patients with colonic cancer (12 men and 8 women, aged 55 to 76 years, mean 62.3 years). A mechanographic technique was used to evaluate in vitro muscle responses to electrical field stimulation (EFS) before and after treatment with various autonomic nerve blockers. Response to EFS before blockade of the adrenergic and cholinergic nerves was as follows: NSC and ESC significantly demonstrated relaxation reaction rather than contraction reaction (P = 0.0253, P < 0.0001, respectively). ESC showed relaxation reaction more than NSC (P = 0.1138). Response to EFS after blockade of the adrenergic and cholinergic nerves was as follows: NSC and ESC significantly demonstrated relaxation reaction via nonadrenergic noncholinergic (NANC) inhibitory nerves rather than contraction reaction via NANC excitatory nerves (P < 0.0001, P < 0.0001, respectively). ESC with SV significantly showed relaxation reaction more than NSC (P = 0.0092). An increased response of relaxation mediated NANC inhibitory nerves may play a role in impaired motility in the ESC of patients with SV.

Previously, we established a pre-operative risk scoring system to predict contralateral inguinal hernia in children with unilateral inguinal hernias. The current study aimed to verify the usefulness of our pre-operative scoring system. This was a prospective study of patients undergoing unilateral inguinal hernia repair from 2006 to 2009 at a single institution. Gender, age at initial operation, birth weight, initial operation side, and the pre-operative risk score were recorded. We analyzed the incidence of contralateral inguinal hernia, risk factors, and the usefulness of our pre-operative risk scoring system. The follow-up period was 36 months. We used forward multiple logistic regression analysis to predict contralateral hernia. Of the 372 patients who underwent unilateral hernia repair, 357 (96.0 %) were completely followed-up for 36 months, and 23 patients (6.4 %) developed a contralateral hernia. Left-sided hernia (OR = 5.5, 95 %, CI = 1.3-24.3, p = 0.023) was associated with an increased risk of contralateral hernia. The following covariates were not associated with contralateral hernia development: gender (p = 0.702), age (p = 0.215), and birth weight (p = 0.301). The pre-operative risk score (cut-off point = 4.5) of the patients with a contralateral hernia was significantly higher, compared with the patients without a contralateral hernia using the area under the receiver operating characteristic curve (p = 0.024). Using multivariate analysis, we confirmed usefulness of our pre-operative scoring system and initial side of the inguinal hernia, together, for the prediction of contralateral inguinal hernia in children.

Aberrant methylation of Nr4a3 exon 3 CpG island (CpGi) was initially identified during multistep mouse skin carcinogenesis. Nr4a3 is also known as a critical gene for neuronal development. Thus, we examined the Nr4a3 exon 3 CpGi methylation in mouse brain tissues from 15-day embryos, newborns and 12-week-old adults and found significant increase of its methylation and Nr4a3 expression during mouse brain development after birth. In addition, homologous region in human genome was frequently and aberrantly methylated in neuroblastoma specimens. A quantitative analysis of DNA methylation revealed that hypomethylation of CpG islands on NR4A3 exon 3, but not on exon 1 was identified in three neuroblastomas compared with matched adrenal glands. Additional analysis for 20 neuroblastoma patients was performed and 8 of 20 showed hypomethylation of the CpGi on NR4A3 exon 3. The survival rate of those 8 patients was significantly lower compared with those in patients with hypermethylation. Immunohistochemical NR4A3 expression was generally faint in neuroblastoma tissues compared with normal tissues. Moreover, the MYCN amplified NB9 cell line showed hypomethylation and low expression of NR4A3, while the non-MYCN amplified NB69 cell line showed hypermethylation and high expression. These results indicate that DNA hypomethylation of the CpGi at NR4A3 exon 3 is associated with low NR4A3 expression, and correlates with poor prognosis of neuroblastoma. Since NR4A3 upregulation associated with the hypermethylation and neuronal differentiation in mice, poor prognosis of neuroblastoma associated with NR4A3 low expression may be partly explained by dysregulation of its differentiation.

Background The present study aimed to assess the long-term results of seton placement for fistula-in-ano (FIA) in infants. Methods Data of patients aged < 1 year who presented to our department with perianal abscess (PA) between January 2006 and February 2010 were retrospectively reviewed. Our standard initial treatment for PA was incision and drainage. Patients with systemic diseases and inflammatory bowel diseases were excluded. Results Ninety-five patients were treated for PA and/or FIA during the 5-year period, and follow-up data were available for 90 patients. The mean follow-up duration in these patients was 49.8 + 11.4 months, and mean age at presentation was 3.1 + 2.7 months. Of the 90 patients, 36 (40 %) developed FIA (39 lesions) and underwent seton placement. The condition healed in a mean period of 6.3 + 4.0 weeks after the placement of a cutting seton. Healing of the fistula was achieved in 35 (97.2%) of 36 patients after the initial seton procedure, and one patient who showed recurrence underwent a second seton placement, resulting in successful healing of the FIA after 5 weeks. Conclusions The long-term success of seton placement indicates that this procedure should be a treatment option for FIA in infants.

小児腸間膜裂孔ヘルニアは、腸閉塞の原因として稀である。我々は、絞扼性イレウスを伴った小児腸間膜裂孔ヘルニアの1例を経験した。症例は、2歳の女児。腸閉塞の診断で当院へ紹介となった。腹部超音波検査とCT検査にて腸内容の貯留を伴った小腸の拡張と腹水を認めた。絞扼性イレウスの診断で緊急開腹術を施行した。小腸間膜に1×2cmの欠損孔を認め、欠損孔に130cmにわたる回腸末端が入り込み壊死していた。小腸壊死部を切除し、欠損孔を修復した。小児腸間膜裂孔ヘルニアについて、文献的考察を含めて報告する。(著者抄録)

We describe the physiological examination of anorectal function in children with chronic constipation (CC), including Hirschsprung's (H) disease (aganglionosis) and its allied disorders [hypoganglionosis (Hypo) and intestinal neuronal dysplasia (IND)]. The assessments are summarized as follows: 1) Anorectal manometry: Enteric nervous system, especially non-adrenergic non-cholinergic excitatory and inhibitory nerves, controls the ano-rectal sphincter reflex. Therefore, patients with aganglionosis lack ano-rectal sphincter reflexes. Abnormalities of the enteric nervous system in Hypo and IND show atypical sphincter reflexes. In contrast, CC patients exhibit normal sphincter reflexes. 2) Terminal motor latency in the pudendal and sacral nerves: Patients with aganglionosis, Hypo, IND and CC exhibit conduction delay of these nerves. These findings may suggest damage to the pudendal and sacral motor nerves due to straining during defecation. 3) Anal canal sensation test (anal canal mucosal electrosensitivity and somatosensory evoked potential tests): Anal sensation is decreased in patients with H disease, Hypo, IND and CC. These findings may suggest damage to the pudendal sensory nerves due to straining during defecation. 4) Defecography: This examination easily detects an obstructive type in school-age patients with CC. 5) Gastrointestinal transit time: This examination easily detects the slow transit type in school-age patients with CC. Conclusions: It is easy for pediatric surgeons to evaluate physiological function in patients with CC, H disease and its allied diseases using physiological examinations.

Background: Laparoscopically assisted anorectoplasty (LAARP) was expected to achieve better fecal continence than conventional procedures. However, the issue of which approach is better remains controversial. We compared outcomes between the conventional procedure and LAARP in male infants with rectoprostatic urethral fistula. Methods: Institutes belonging to the Japanese Study Group of Anorectal Anomalies (JSGA) were invited to participate. Subjects were male infants with rectoprostatic urethral fistula treated by the conventional approach (abdominoperineal pull-through and PSARP) or LAARP between 2000 and 2006. Medical charts and operative records were reviewed retrospectively. Results: Eighty-one patients (conventional: 36, LAARP: 45) were enrolled from 15 centers. In both groups, the mean Kelly score was 5. The total score of the scoring system was newly developed by the Japanese Study Group of Anorectal Anomalies. Follow-up Project (5-15 points) was 10.7 and 12.1 in the conventional group and the LAARP group, respectively (p = 0.07). The incidence of failed rectoanal anastomosis, mucosal prolapse, and anal stenosis was comparable in both groups. Posterior urethral diverticula were detected on cystourethrograms in 7% and 11% (p = 1.0) and on MRI in 0% and 34% (p = 0.02) of the conventional and the LAARP groups, respectively. Overall, 94% of diverticula were asymptomatic. Conclusions: Fecal continence and complication rates after LAARP were comparable to those observed after the conventional method. Posterior urethral diverticula were detected more frequently after LAARP. © 2013 Elsevier Inc. All rights reserved.

Abstract We present the case of a 13-year-old patient who underwent right salpingo-oophorectomy and was diagnosed with stage Ia ovarian mucinous cystadenocarcinoma. Nine years after complete resection, she was diagnosed with metastasis of the previously resected carcinoma. Despite receiving combination chemotherapy, the patient died of the disease. Long-term follow-up of teenage patients with stage Ia ovarian mucinous cystadenocarcinoma is important. © 2013 Elsevier Inc. All rights reserved.

Background: The comprehensive methylation analysis of tumor-specific differently methylated regions in malignant melanomas and brain tumors has led to the identification of non-promoter hypermethylation of zygote arrest 1 (ZAR1). To search the non-promoter ZAR1 hypermethylation in neuroblastomas, we analyzed the levels of the methylation and transcript expression of ZAR1. Methods: The MassARRAY (R) EpiTYPER (Sequenom Inc., San Diego, CA, USA) system was optimized to determine the quantitative methylation levels of ZAR1 for 12 neuroblastoma cell lines, 23 neuroblastoma samples and four adrenal samples. ZAR1 expression levels were evaluated through a quantitative, real-time reverse transcription-polymerase chain reaction. The quantitative methylation levels of ZAR1 were subjected to correlation studies with the established markers of progressive disease and outcome. Results: Strikingly, the hypermethylation of ZAR1 regions and ZAR1 expression levels was observed in the neuroblastoma cell lines and neuroblastoma samples, compared to the adrenal samples. Somatic changes in ZAR1 methylation and ZAR1 expression were found in all three neuroblastoma patients. In the ZAR1 regions, poor-outcome tumors that were MYCN-amplified and/or Stage 3 or 4 and/or the age at diagnosis was >= 18 months, and/or showed an unfavorable histology were frequently hypermethylated. Conclusion: Our results indicate that the hypermethylation of ZAR1 regions is extremely frequent in neuroblastomas and correlates with established markers of progressive disease and outcome. (c) 2013 Elsevier Inc. All rights reserved.

Background The identification of tissue-specific differentially methylated regions (tDMRs) is key to our understanding of mammalian development. Research has indicated that tDMRs are aberrantly methylated in cancer and may affect the oncogenic process. Procedure We used the MassARRAY EpiTYPER system to determine the quantitative methylation levels of seven neuroblastomas (NBs) and two control adrenal medullas at 12 conserved tDMRs. A second sample set of 19 NBs was also analyzed. Statistical analysis was carried out to determine the relationship of the quantitative methylation levels to other prognostic factors in these sample sets. Results Screening of 12 tDMRs revealed 2 genomic regions (SLC16A5 and ZNF206) with frequent aberrant methylation patterns in NB. The methylation levels of SLC16A5 and ZNF206 were low compared to the control adrenal medullas. The SLC16A5 methylation level (cut-off point, 13.25%) was associated with age at diagnosis, disease stage, and Shimada classification but not with MYCN amplification. The ZNF206 methylation level (cut-off point, 68.80%) was associated with all of the prognostic factors analyzed. Although the methylation levels at these regions did not reach statistical significance in their association with prognosis in mono-variant analysis, patients with both hypomethylation of SLC16A5 and hypermethylation of ZNF206 had a significantly prolonged event-free survival, when these two variables were analyzed together. Conclusions We demonstrated that two tDMRs frequently displayed altered methylation patterns in the NB genome, suggesting their distinct involvement in NB development/differentiation. The combined analysis of these two regions could serve as a diagnostic biomarker for poor clinical outcome. Pediatr Blood Cancer 2013; 60: 383-389. (C) 2012 Wiley Periodicals, Inc.