The effect of laminin-1 on enteric neural crest-derived cell migration in the Hirschsprung's disease mouse model.

BACKGROUND/AIM: Laminin-1 regulates neurite outgrowth in various neuronal cells. We have previously demonstrated that laminin-1 promotes enteric neural crest-derived cell (ENCC) migration by using Sox10-VENUS transgenic mice, in which ENCCs are labeled with a green fluorescent protein, Venus. Mice lacking the endothelin-B receptor gene, Ednrb -/- mice, are widely used as a model for Hirschsprung's disease (HD). The aim of this study was to investigate the effects of laminin-1on ENCC migration in Sox10-VENUS+/Ednrb -/- mice, a newly created HD mice model. METHODS: Fetal guts were dissected on embryonic day 12.5 (E12.5). Specimens were incubated either with, or without laminin-1 for 24 h and images were taken under a stereoscopic microscope. The length from the stomach to the wavefront of ENCC migration (L-E) and the total length of the gut (L-G) were measured. Changes in the ratio of L-E to L-G (L-E/L-G) after 24 h were calculated. RESULTS: On E12.5, the wavefront of ENCC migration in the HD gut samples was located in the midgut, whereas the wavefront of ENCC in Sox10-VENUS+/Ednrb +/+ (WT) samples had reached the hindgut. After 24 h, L-E/L-G had increased by 1.49%, from 34.97 to 36.46%, in HD gut and had increased by 1.07%, from 48.08 to 49.15%, in HD with laminin-1, suggesting there was no positive effect of laminin-1 administration on ENCC migration in HD. CONCLUSIONS: Our results suggest that laminin-1 does not have a positive effect on ENCC migration in HD mice on E12.5, in contrast to the phenomenon seen in normal mice gut specimens, where laminin-1 promotes ENCC migration during the same period. This suggests that there is an impairment in the interaction between ENCC and extracellular environmental factors, which are required for normal development of the enteric nervous system, resulting in an aganglionic colon in HD.

Droplet digital PCR measurement of HER2 in patients with gastric cancer.

BACKGROUND: Although there are some new criteria for human epidermal growth factor receptor 2 (HER2) expression with immunohistochemistry/fluorescence in situ hybridisation (IHC/FISH) in gastric cancer, the method is still ambiguous and is somewhat dependent on the subjective qualities of the evaluator. METHODS: We used droplet digital polymerase chain reaction (ddPCR) to evaluate HER2 amplification in formalin-fixed and paraffin-embedded (FFPE) samples and cell-free serum circulating tumour DNA (ctDNA) in 25 patients with gastric cancer. RESULTS: The concordance rate of HER2 amplification examined in FFPE samples with ddPCR and IHC/FISH was 92% (23 out of 25). The concordance rate of FFPE with ctDNA was not high (62.5%); however, patients who were HER2-positive by ctDNA had significantly shorter survival compared with HER2-negative patients. CONCLUSIONS: Our results demonstrated that this ddPCR method was as effective as IHC/FISH and therefore might become a standard method for analysing not only FFPE but also ctDNA.

Effect of hepatic arterial infusion chemotherapy of 5-fluorouracil and cisplatin for advanced hepatocellular carcinoma in the Nationwide Survey of Primary Liver Cancer in Japan.

BACKGROUND: The efficacy of hepatic arterial infusion chemotherapy for the treatment of advanced hepatocellular carcinoma (HCC) remains unclear. METHODS: The outcome of 476 patients with HCC who underwent hepatic arterial infusion chemotherapy with 5-fluorouracil and cisplatin (HAIC) were compared with 1466 patients who did not receive active therapy. RESULTS: A survival benefit of the therapy after adjusting for known risk factors was observed (hazard ratio, 0.48; 95% CI, 0.41-0.56; P<0.0001). In propensity score-matched analysis (n=682), median survival time was longer for patients who underwent chemotherapy (14.0 months) than for patients who did not receive active treatment (5.2 months, P<0.0001). CONCLUSION: For advanced HCC, HAIC is considered to be an effective treatment.

Pro-angiogenic cytokines for prediction of outcomes in patients with advanced hepatocellular carcinoma.

BACKGROUND: We previously reported that expressions of the pro-angiogenic cytokines angiopoietin-2 (Ang-2), follistatin, granulocyte colony-stimulating factor, hepatocyte growth factor, leptin, platelet-derived growth factor-BB, platelet endothelial cell adhesion molecule-1, and vascular endothelial growth factor were associated with the response to sorafenib in patients with advanced hepatocellular carcinoma (HCC). The aim of the present study is to examine the same relationship in a larger cohort. METHODS: In the current retrospective cohort study, we measured serum levels of the eight cytokines in 120 consecutive HCC patients who were treated with sorafenib. We evaluated the effects of increased expression of serum cytokines on progression-free survival (PFS) and overall survival (OS). RESULTS: Elevated expression of Ang-2 correlated both with significantly shorter PFS (hazard ratio (HR), 1.84; 95% confidence interval (CI), 1.21-2.81), and OS (HR, 1.95; 95% CI, 1.21-3.17). Patients with more than three cytokines expressed above the median similarly had significantly shorter PFS (HR, 1.98; 95% CI, 1.30-3.06) and OS (HR, 1.94; 95% CI, 1.19-3.22). Differences in OS were evident in cases with the evidence of macroscopic vascular invasion or extrahepatic metastasis. CONCLUSION: High expression of Ang-2 or more than cytokines in serum is associated with poor PFS and OS in HCC patients treated with sorafenib.

Role of interleukin-17A in the eosinophil accumulation and mucosal remodeling in chronic rhinosinusitis with nasal polyps associated with asthma.

BACKGROUND: Interleukin (IL)-17A is a highly inflammatory cytokine with a robust effect on stromal cells in many tissues. Although IL-17A is known to be associated with inflammatory lung disorders by triggering an accumulation of neutrophils, the effect of IL-17A on the upper airway is still uncertain. The expression of IL-17A and its role were investigated in the nasal polyps of chronic rhinosinusitis associated with asthma. METHODS: IL-17A was detected by immunohistochemistry and quantitative real-time RT-PCR. The cellular source of IL-17A was examined by double staining with EG2, CD4 and neutrophil elastase. The tissue remodeling of the nasal polyps was evaluated by assessing the epithelial damage and basement membrane thickness. RESULTS: Both the immunoreactivity and mRNA of IL-17A were significantly detected in the nasal polyps in comparison with control normal sinus mucosa. The localization of IL-17A expression predominantly coincided with eosinophils and CD4-positive lymphocytes. Furthermore, the number of IL-17A-positive cells correlated with tissue eosinophils, but not with neutrophils. The degree of epithelial damage and basement membrane thickness was dependent on the number of infiltrated IL-17A-positive cells. CONCLUSION: The present study suggests, for the first time, that IL-17A plays an important role in the eosinophil accumulation in the nasal polyps and the remodeling of the nasal polyps of chronic rhinosinusitis associated with asthma.

Relationship between epithelial damage or basement membrane thickness and eosinophilic infiltration in nasal polyps with chronic rhinosinusitis.

BACKGROUND: Chronic rhinosinusitis (CRS) with nasal polyps is characterized by eosinophilic infiltration. This study hypothesized that the aggregation of the mucosal pathology during remodeling is related to infiltrating eosinophils in patients with such nasal polyps. OBJECT: To clarify the pathogenetic role of eosinophils in patients with CRS with nasal polyps, this study investigated the relationship between epithelial damage or basement membrane (BM) thickening and the epithelial infiltration of eosinophils in these nasal polyps. METHODS: The number of eosinophils that infiltrated into the epithelial and subepithelial layers of sinonasal tissues was counted. The staging of epithelial damage allowed the quantification of epithelial loss. RESULTS: Both epithelial damage and BM thickness in CRS, which were correlated with the number of infiltrated eosinophils, were significantly greater than in the control group. Neither parameter showed significant differences between the asthma and non-asthma groups. There was a significantly correlation in the eosinophilic infiltration between the subepithelial and epithelial layers. CONCLUSION: It is suggested that eosinophils that infiltrate into both the epithelial and subepithelial layers play a part in the process of mucosal remodeling of CRS with nasal polyps.

[New method for preparing proximal anastomotic system].

Heartstring is a useful device. However, the device failure at the time of loading the seal into the delivery device is a troublesome issue. To avoid this problem, we invent a new method using 2 tourniquets made of 5 mm-wide woven Teflon tapes and plastic tubes. Using our method, the loading procedure became easier and more reliable.

[Successful treatment for tachyarrhythmia with ultra short acting beta-blocker during beating coronary artery bypass grafting; report of a case].

A 62-year-old man with supraventricular/ventricular tachycardia associated with myocardial infarction developed tachycardia during beating coronary artery bypass grafting (CABG). Intravenous administration of an ultra short acting beta-blocker, landiolol hydrochloride, controlled heart rate and improved tachyarrhythmia without significant change of blood pressure. Landiolol hydrochloride is effective and useful for the treatment of tachyarrhythmia during beating CABG.

Total aortic arch graft replacement for recurrent aortic aneurysm.

We successfully performed a total aortic arch replacement for a recurrent aortic aneurysm following repair of an aortic dissection. A 59-year-old man underwent a patch aortoplasty through median sternotomy for Stanford type B aortic dissection in other hospital. Three years and 6 months later an aneurysm developed. Computed tomography and magnetic resonance imaging angiography demonstrated an enlargement of the aneurysm, resulting in a diagnosis of recurrent distal aortic arch aneurysm. A graft replacement of the total aortic arch with the aid of selective cerebral perfusion was performed through a median resternotomy and left lateral thoracotomy. Additional left lateral thoracotomy offered a sufficiently optimal operating field for distal anastomosis. However, care must be taken not to overlook the bleeding from intercostal arteries. Since aortoplasty may lead to subsequent dilation and aneurysmal formation, initial replacement of the segment of the aorta is recommended, and careful long-term follow-up of the patient is important.

[Extraction of bisphenol-A from a cardiopulmonary bypass circuit].

We measured levels of bisphenol-A (BPA) in a priming solution and blood of a cardiopulmonary bypass (CPB) circuit. Eight circuits were used in the study of a priming solution. Blood samples were obtained from 6 patients who underwent open heart surgery after the commencement of CPB and at the termination of CPB. Another 3 samples were collected directly from the saline in a polyethylene container as a control. Then the concentrations of BPA in them were determined by means of gas chromatography. No detectable BPA was found in controls. However, a small amount was detected in the saline from the circuits (0.9 +/- 1.1 micrograms/l). A very small amount was also detected in the blood after the commencement and at the termination of CPB (0.3 +/- 0.2 microgram/l, and 0.4 +/- 0.3 microgram/l, respectively). BPA was considered to be leached from the circuit to the priming solution and the blood because the parts of the reservoir and the oxygenator were made of polycarbonate containing BPA. We suppose the BPA concentration is probably at a safe level. However, the data on the endocrinologically toxic level of blood BPA are insufficient. Therefore, the use of plastic in a circuit may require closer scrutiny to determine whether BPA contributes to exposure to xenoestrogens.

[Repair of tetralogy of Fallot in an adult; the importance of preoperative examination for major aorto-pulmonary collateral arteries].

We report the repair of tetralogy of Fallot in a 51-year-old man. He underwent a left classical Blalock-Taussig (B-T) shunt when he was ten years old. Preoperative angiography showed a patent B-T shunt. Total corrective surgery was performed. Postoperatively, he suffered from left ventricular failure. Re-intubation was required three times. Postoperative catheterization showed excellent correction of the right ventricular system, however, descending aortography revealed a great many aorto-pulmonary collateral arteries, which caused the left ventricular failure. Coil embolization of these 13 collateral arteries was performed and he recovered from the left ventricular failure. We conclude that even in an elderly patient with tetralogy of Fallot, total correction should be performed, since the surgical risk is acceptable and the procedure improves the capacity for physical activity and quality of life. Preoperative examination of collateral arteries is important, especially in elderly patients, and coil embolization should be considered.

[Successful treatment of a patient with low pressure syndrome associated with gait disturbance].

The authors report an unusual case of low intracranial pressure (ICP) syndrome that was successfully treated by the placement of an anti-siphon device (ASD). This 36-year-old male had suffered suprasellar germinoma with hydrocephalus and had had a V-P shunt following radiotherapy. Sixteen years later he developed gait disturbance and somnolence and MRI demonstrated a small lateral ventricle as well as a diffuse dural enhancement. A lumbar tap revealed a low ICP of 12 mmH2O. Because of this, an ASD was placed in the patient. Postoperatively, his symptoms of gait and consciousness disturbance improved. Typical clinical findings of low ICP syndrome such as headache were not observed in this case. To our knowledge, no symptom of gait disturbance with low ICP has been reported previously. We present an interesting case of low ICP syndrome with gait disturbance and discuss the mechanism of the symptoms. Symptoms of this patient were due at first to brain ischemia. After convulsion and consciousness disturbance due to low intracranial pressure, the symptoms increased in strength until gait disturbance occurred. The possibility is suggested that gait disturbance in this patient was due both to brain ischemia and low intracranial pressure.

[Surgical reconstruction for radiation-induced extracranial vertebral artery stenosis: a case report].

We report a case of symptomatic extracranial vertebral artery stenosis after radiation therapy. This 49-year-old female received radiation therapy to the neck for nasopharyngeal carcinoma 11 years earlier, was admitted because of continuous dizziness and a floating sensation. Magnetic resonanse imaging showed no abnormalities, but an aortography demonstrated complete occlusion of the right common carotid artery as well as occlusion of the right vertebral artery and severe stenosis of the left vertebral artery at its origin, which was presumed to be the result of previous radiation therapy. Percutaneous transluminal angioplasty (PTA) for the left vertebral artery was performed using conventional balloon treatment, which resulted in wall dissection. Because of this, she underwent end-to-side vertebral artery to subclavian artery transposition, and she has had no further ischemic events science that time. PTA has been successfully performed as the first treatment of choice for vertebral artery stenosis, but surgical reconstruction can be a therapeutic management of choice for cases of failed PTA.

[A case of diffuse alveolar hemorrhage developing after open heart surgery].

A 47-year-old man was receiving anticoagulant therapy after coronary artery bypass grafting and mitral valve plasty. A chest roentgenogram disclosed alveolar infiltrates throughout both lung fields 6 months postoperatively. Lung biopsy specimens showed the presence of hemosiderin-laden macrophages in the alveolar spaces, establishing a diagnosis of diffuse alveolar hemorrhage. Anticoagulant therapy may have caused the hemorrhage, because the patient had no immunologic disorder or renal disease. The shadows completely disappeared after steroid therapy and discontinuation of anticoagulant therapy. Diffuse alveolar hemorrhage should be considered, when extensive infiltrates develop on chest roentgenogram in patients receiving anticoagulant therapy after open heart surgery. Lung biopsy is essential, when a bronchoalveolar lavage fails to diagnose the disease.

[Reoperation for late postoperative right ventricular outflow tract obstruction].

Between February 1993 and July 2000, 18 patients with a mean age of 7.9 years underwent re-operation for critical stenosis of the right ventricular outflow tract. Their diagnoses of these patients included tetralogy of Fallot (n = 7), transposition of the great arteries (n = 6), truncus arteriosus communis (n = 3), and double outlet right ventricle with pulmonary stenosis (n = 2). The first operations were extracardiac conduit operations (n = 9), arterial switch operations (n = 6) and patch reconstruction of the right ventricular outflow tract (n = 3). At re-operation, the right ventricular outflow tract obstruction was released completely and reconstructed using a monocusp ventricular outflow patch (n = 14), valved pericardial roll (n = 2), or bovine pericardial patch sparing the native pulmonary valve annulus (n = 2). There were no early deaths, although one patient died of arrhythmia 2.6 years after re-operation. The pressure gradient across the right ventricular outflow tract, right ventricular systolic pressure, and right ventricle to systemic systolic pressure ratio were satisfactorily relieved (84.3 +/- 19.0 vs. 16.7 +/- 19.7 mmHg, 109.2 +/- 20.5 vs. 48.7 +/- 16.7 mmHg, 0.92 +/- 0.25 vs. 0.43 +/- 0.16; p < 0.001, p < 0.001, p < 0.001, respectively). The right ventricular end-diastolic volume index and right ventricular ejection fraction were also improved (166.3 +/- 85.2 vs. 105.6 +/- 28.8% of normal, 38.3 +/- 12.6 vs. 50.9 +/- 8.8%; p < 0.05, p < 0.005, respectively). Re-operation should be done before the development of marked right ventricular dysfunction, while it can still be performed at a low risk with satisfactory results.

Transplantation of newborn esophagus: an experimental study.

PURPOSE: The aim of this study was to see if allogeneic transplantation (Tx) of newborn esophagus can create viable esophageal tissue that may be used for treating long gap esophageal atresia. METHODS: Specimens of thoracic esophagus from newborn Brown-Norway rats, each were transplanted into a pouch created in the distal omentum of 5-week-old Lewis rats. In group I no immunosuppressant was used. FK-506 was used in group II (0.2 mg/kg), group III (0.6 mg/kg), and group IV (1.2 mg/kg) until a predetermined day of graft harvesting (1, 2, 3, 4, 5, 6, and 8 weeks after Tx). FK-506 was used for only 2 weeks in group V (0.6 mg/kg), and group VI (1.2mg/kg), and transplanted esophageal grafts were harvested 1, 2, 3, and 4 weeks after cessation of 2 weeks course FK-506. Syngeneic esophagus transplants were used as controls. All grafts were examined by H&E staining to assess graft viability and degree of rejection. RESULTS: Each successfully transplanted esophagus appeared macroscopically as a tube like mass. Each graft could be mobilized to the thoracic cavity, because of the long omental pedicle. Graft survival in the control group was 100%. Rejection was observed in all grafts from groups I, II, V, and VI. In contrast, grafts from groups III and IV showed only minimal or no rejection. There was no evidence of side effects of FK-506 in rats in groups III and IV, except significantly slower weight gain compared with controls (P <.05). CONCLUSIONS: FK-506 successfully prevented rejection, although immunologic tolerance was not achieved. These observations suggest that the authors' procedure has the potential to produce viable esophageal tissue that could be a new option for treating long gap esophageal atresia.

Pyloric stenosis: new histopathologic perspective using confocal laser scanning.

BACKGROUND/PURPOSE: Idiopathic hypertrophic pyloric stenosis (IHPS) is a common infantile disorder characterized by enlargement of the pylorus and gastric outlet obstruction. Its complete etiology is still not fully understood, but recent research has focussed on abnormalities of nerve distribution. The authors used confocal laser scanning microscopy to perform 3-dimensional studies of pylorus biopsy specimens taken from cases of IHPS and present their findings. METHODS: Pylorus biopsy specimens obtained at pyloromyotomy from 6 infants with IHPS were studied using confocal microscopy and compared with 6 control pylorus biopsy specimens from patients without gastrointestinal disease. Biopsy specimens were pretreated to enhance nerve expression by using protein gene product 9.5 (PGP9.5) polyclonal antibody to identify enteric nerve system fibers. Double staining immunofluorescence was used to detect alpha smooth muscle actin (SMA), a smooth muscle marker. RESULTS: Control pylorus biopsy specimens showed many thin PGP9.5-positive nerve fibers in the circular and longitudinal muscle layers that communicated with each other to create a 3-dimensional meshlike network. Muscle cells stained by alpha SMA antibody were thin. In contrast, muscle cells from IHPS patients were fat and round. The PGP9.5 staining nerve fibers from IHPS patients formed numerous, thick, and contorted bundles that did not communicate. CONCLUSIONS: By using confocal laser microscopy the authors were able to identify abnormally thick contorted nerve bundles in the pyloric muscle layers of infants with IHPS. These anormal nerve bundles have not been described previously because of the limitations of 2-dimensional microscopy. The authors suspect that the etiology of IHPS may be related to these abnormal fibers.

Intestinal neuronal dysplasia-like pathology in Ncx/Hox11L.1 gene-deficient mice.

BACKGROUND/PURPOSE: Ncx/Hox11L.1-deficient (Ncx-/-) mice specifically created by the authors had mega-ileo-ceco-colon (mega-ICC) with a caliber change in the proximal colon. The authors studied the nerve distribution in the bowel of these Ncx-/- mice to determine the cause of their bowel dysmotility. METHODS: Four-week-old Ncx-/- mice (n = 10; 5 with mega-ICC, 5 without mega-ICC) were killed and the bowel harvested. Half of each specimen was snap frozen for AchE and NADPH-diaphorase histochemistry, and the other half were fixed with 10% formalin for H&E staining and immunohistochemistry using PGP9.5 antibody (a marker for neurons), C-kit antibody (a marker for intestinal pacemaker cells), and stem cell factor antibody (a marker for C-kit ligand). Age-matched wild-type normal mice (n = 5) served as controls. RESULTS: In the ileum, cecum, and proximal colon from all Ncx-/- mice (irrespective of the association of mega-ICC), typical findings of human intestinal neuronal dysplasia (IND) ie, obvious hyperganglionosis in neuronal plexuses on PGP9.5 immunohistochemistry, ectopic ganglia in the mucosal and muscular layers on AchE histochemistry, and ghostlike ganglia on NADPH-diaphorase histochemistry were found. Likewise, in normal caliber distal colon from these mice, the distribution of ganglion cells, C-kit, and stem cell factor was normal. In control specimens, there was no ectopic ganglia or hyperganglionosis. CONCLUSIONS: These findings suggest that the Ncx/Hox11L.1 gene is required for the proper innervation of the enteric nervous system in mice, and our deficient strain may be useful as a model for studying IND in humans.

Bladder transplantation in rats using FK-506.

PURPOSE: We created viable bladder tissue by transplantation with immunosuppression. MATERIALS AND METHODS: For bladder transplantation the bladder of newborn Brown-Norway rats was excised and each was transplanted into a pouch created in the distal omentum of a 5-week-old Lewis rat. In 15 group 1 rats no immunosuppressive agent was used. In 20 group 2 rats 0.6 mg./kg. FK-506 daily were given intramuscularly until a predetermined day of harvest. Recipient rats were sacrificed on day 3, 5, 7 or 14 after bladder transplantation, and the bladder grafts were harvested and formalin fixed. Hematoxylin and eosin staining was done to examine bladder graft survival and the degree of rejection, and immunohistochemical testing was performed for assessing the vesical nervous system. In 5 rats in the control group bladder augmentation was performed by anastomosing the bladder graft to the native bladder. Each augmented bladder was harvested 21 days later for histopathological assessment. RESULTS: Overall bladder graft survival was 96.4%. Each successfully transplanted bladder graft appeared macroscopically as a thin walled cyst. In group 1 all bladder grafts showed rejection with cellular infiltration. In group 2 there was mild rejection in 5 rats and no evidence of rejection in the remaining 15. All group 2 bladder grafts had intact nerve distribution. Bladder augmentation was successful in all 5 cases and the mucosa was normal throughout each augmented bladder. CONCLUSION: Because FK-506 successfully prevents rejection, our technique would appear to have the potential for creating viable bladder tissue that may be used for bladder augmentation in cases of vesical exstrophy or neurogenic bladder.