Functional roles of CD26/DPP4 in lipopolysaccharide-induced lung injury.

Acute respiratory distress syndrome (ARDS) is characterized by dysregulated inflammation and increased permeability of lung microvascular cells. CD26/dipeptidyl peptidase-4 (DPP4) is a type II membrane protein that is expressed in several cell types and mediates multiple pleiotropic effects. We previously reported that DPP4 inhibition by sitagliptin attenuates lipopolysaccharide (LPS)-induced lung injury in mice. The current study characterized the functional role of CD26/DPP4 expression in LPS-induced lung injury in mice, isolated alveolar macrophages, and cultured lung endothelial cells. In LPS-induced lung injury, inflammatory responses [bronchoalveolar lavage fluid (BALF) neutrophil numbers and several proinflammatory cytokine levels] were attenuated in Dpp4 knockout (Dpp4 KO) mice. However, multiple assays of alveolar capillary permeability were similar between the Dpp4 KO and wild-type mice. TNF-α and IL-6 production was suppressed in alveolar macrophages isolated from Dpp4 KO mice. In contrast, in cultured mouse lung microvascular endothelial cells (MLMVECs), reduction in CD26/DPP4 expression by siRNA resulted in greater ICAM-1 and IL-6 expression after LPS stimulation. Moreover, the LPS-induced vascular monolayer permeability in vitro was higher in MLMVECs treated with Dpp4 siRNA, suggesting that CD26/DPP4 plays a protective role in endothelial barrier function. In summary, this study demonstrated that genetic deficiency of Dpp4 attenuates inflammatory responses but not permeability in LPS-induced lung injury in mice, potentially through differential functional roles of CD26/DPP4 expression in resident cellular components of the lung. CD26/DPP4 may be a potential therapeutic target for ARDS and warrants further exploration to precisely identify the multiple functional effects of CD26/DPP4 in ARDS pathophysiology.NEW & NOTEWORTHY We aimed to clarify the functional roles of CD26/DPP4 in ARDS pathophysiology using Dpp4-deficient mice and siRNA reduction techniques in cultured lung cells. Our results suggest that CD26/DPP4 expression plays a proinflammatory role in alveolar macrophages while also playing a protective role in the endothelial barrier. Dpp4 genetic deficiency attenuates inflammatory responses but not permeability in LPS-induced lung injury in mice, potentially through differential roles of CD26/DPP4 expression in the resident cellular components of the lung.

Development of a dye nanoparticle-loaded test strip for the Zn(II) determination in metal plating wastes.

Zincon/Latex-NR3+ nanocomposite-loaded dye nanoparticle coated test strip (Zincon/Latex-NR3+ DNTS) was fabricated to detect Zn(II) in plating wastewater by a unique color change from red-purple to deep blue and evaluated its detection performance in actual plating samples. The 5 × 5 mm square-cut DNTS attached sticks were immersed into 10 mL aliquots of aqueous solutions containing Zn(II) ion and 0.01 M TAPS buffer at pH 8.4 with stirring at 250 rpm for 60 min. The calibration curve for Zn(II) was prepared based on the integrated area intensity of reflectance by TLC at 620 nm with the 3σ detection limit was 48.61 ppb, and the quantitative range was approximately up to 1000 ppb. Although Cu(II), Mn(II), Ni(II), and Co(II) showed competitive interference due to complex formation with Zincon, a mixture of masking reagent including thiourea, 2-aminoethanthiol, and o-phenanthroline was effective in removing the contamination. To eliminate Cr(III) interference based on the incorporation of Zn(II) into Cr(III) hydrolyzed polymer, adding KBrO3 and H2SO4 under boiling for several minutes was required. With appropriate pretreatment, all results of actual plating water samples by Zincon/LatexNR3+ DNTS were close to those of ICP-OES.

Functional roles of CD26/DPP4 in bleomycin-induced pulmonary fibrosis.

The pathogenesis of pulmonary fibrosis involves complex interplay between cell types and signaling pathways. Recurrent alveolar epithelial injury can occur during pulmonary inflammation, causing dysregulation of epithelial repair. Dysregulated repair interacts with mesenchymal, inflammatory, and endothelial cells to trigger fibroblast-to-myofibroblast activation. CD26/dipeptidyl peptidase-4 (DPP4) is a type II membrane protein mediating pleiotropic effect. However, the mechanistic role of CD26/DPP4 in pulmonary fibrosis remains unclear. In this study, we aimed to characterize Dpp4 deficiency in a mouse bleomycin (BLM)-induced pulmonary fibrosis model and in cell culture systems of human lung fibroblasts (HLFs). Dpp4 knockout (Dpp4 KO) mouse lungs exhibited lower Ashcroft scale indices, collagen content, and numbers of fibroblasts and myofibroblasts compared with those in C57BL/6 wild-type (WT) mice. Upregulation of Tgfb1 and Tgfb2 mRNA levels in the lungs after BLM treatment was lower in Dpp4 KO mice compared with those in WT mice. Although TGF-ß-driven endothelial-to-mesenchymal transition (EndMT) has been implicated as one of the mechanisms of pulmonary fibrosis, a number of partial EndMT cells in lungs did not differ between Dpp4 KO mice and WT mice. The proliferation capacity and mRNA levels of COL1A1, a collagen deposition-related gene, in cultured HLFs were suppressed in DPP4 small interfering RNA-treated cells. This study indicates that the genetic deficiency of DPP4 has protective effects against BLM-induced pulmonary fibrosis, partly through the reduction in TGF-ß expression and inhibition of fibroblast activation in the lung. Our study suggests that CD26/DPP4 inhibition is a potential therapeutic strategy for pulmonary fibrosis.

Functional Roles for CD26/DPP4 in Mediating Inflammatory Responses of Pulmonary Vascular Endothelial Cells.

Excessive inflammation in the lung is a primary cause of acute respiratory distress syndrome (ARDS). CD26/dipeptidyl peptidase-4 (DPP4) is a transmembrane protein that is expressed in various cell types and exerts multiple pleiotropic effects. We recently reported that pharmacological CD26/DPP4 inhibition ameliorated lipopolysaccharide (LPS)-induced lung injury in mice and exerted anti-inflammatory effects on human lung microvascular endothelial cells (HLMVECs), in vitro. However, the mechanistic roles of CD26/DPP4 in lung injury and its effects on HLMVECs remain unclear. In this study, transcriptome analysis, followed by various confirmation experiments using siRNA in cultured HLMVECs, are performed to evaluate the role of CD26/DPP4 in response to the pro-inflammatory involved in inflammation, barrier function, and regenerative processes in HLMVECs after pro-inflammatory stimulation. These are all functions that are closely related to the pathophysiology and repair process of lung injury. Confirmatory experiments using flow cytometry; enzyme-linked immunosorbent assay; quantitative polymerase chain reaction; dextran permeability assay; WST-8 assay; wound healing assay; and tube formation assay, reveal that the reduction of CD26/DPP4 via siRNA is associated with altered parameters of inflammation, barrier function, and the regenerative processes in HLMVECs. Thus, CD26/DPP4 can play a pathological role in mediating injury in pulmonary endothelial cells. CD26/DPP4 inhibition can be a new therapeutic strategy for inflammatory lung diseases, involving pulmonary vascular damage.

[Efficacy and Safety of Gefitinib as First-Line Chemotherapy for Elderly Patients with Advanced Non-Small Cell Lung Cancer(NSCLC)].

Epidermalgrowth factor receptor tyrosine kinase inhibitor(EGFR-TKI)is the first choice for the treatment of EGFR mutation- positive advanced non-small cell lung cancer(NSCLC). There have been few reports on the efficacy and safety of gefitinib in elderly patients with EGFR mutation-positive advanced NSCLC. We retrospectively assessed the efficacy and safety of gefitinib as first-line chemotherapy in 22 patients with advanced NSCLC aged 75 years or older and who were treated with gefitinib. The response rate was 81.8%, and the disease controlrate was 95.5%. The median progression-free survivaltime was 14.2 months, and the median survivaltime was 30.7 months. The common adverse events were skin toxicities(50.0%), liver dysfunction(18.2%), and diarrhea(18.2%). The dose of gefitinib was reduced in 36.3% of the patients, and the treatment of gefitinib was discontinued in 18.2% of the patients. Gefitinib is effective and safe for elderly patients with advanced NSCLC.

The expression of 11 beta hydroxysteroid dehydrogenase in severe allergic rhinitis.

OBJECTIVE: To clarify the roles of 11 beta-HSD in resistance to glucocorticoid therapy for allergic rhinitis, a case series study was conducted. METHODS: The patient group consisted of 20 subjects with allergic rhinitis, aged from 21 to 46 years (mean age 26.5), who showed persistent GC resistance necessitating surgical removal of the inferior turbinate after 6 months' GC treatment. The patients with poor response to GC treatment for 6 months' were defined as GC resistance. The control group consisted of 10 subjects aged from 16 to 39 years (mean age 24.5) who underwent maxillofacial surgery, from whom nasal tissues were taken and who did not receive GC treatment. Nasal mucosal tissues from patients and cntorol subjects were examined immunohistochemically. The sections were washed with 0.01 M phosphate-buffered saline (PBS; pH 7.2) containing 0.15 M NaCl and 0.01% Triton X-100, and incubated for 2 h with rabbit polyclonal anti-11 beta HSD1 and 11 beta-HSD2 antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), each diluted 1:200 in PBS containing 0.1% bovine serum albumin. Immunostained sections were assessed under an Olympus microscope with an eyepiece reticule at 200 X magnification. Cell counts are expressed as means per high-power field (0.202 mm2). Control group means (arithmetic mean ± SD) were compared with patient group means by Mann-Whitney U-test at P = 0.05. RESULTS: Although 11 beta-HSD1 was expressed to a similar extent in patients and controls, 11 beta-HSD2 was expressed significantly more in patients with severe allergic rhinitis, resulting in a increased HSD-1/HSD-2 ratio. The significantly increased expression of 11 beta-HSD2 in the nasal epithelium and submucosal inflammatory cells of patients with severe nasal allergy were observed in the present study. CONCLUSION: Our findings suggest that 11 beta-HSD2 plays an important role in resistance to glucocorticoid therapy for allergic rhinitis, and its expression might be used as an additional parameter indicating steroid resistance in allergic rhinitis.

Thromboembolic adverse event study of combined estrogen-progestin preparations using Japanese Adverse Drug Event Report database.

Combined estrogen-progestin preparations (CEPs) are associated with thromboembolic (TE) side effects. The aim of this study was to evaluate the incidence of TE using the Japanese Adverse Drug Event Report (JADER) database. Adverse events recorded from April 2004 to November 2014 in the JADER database were obtained from the Pharmaceuticals and Medical Devices Agency (PMDA) website (www.pmda.go.jp). We calculated the reporting odds ratios (RORs) of suspected CEPs, analyzed the time-to-onset profile, and assessed the hazard type using Weibull shape parameter (WSP). Furthermore, we used the applied association rule mining technique to discover undetected relationships such as the possible risk factors. The total number of reported cases in the JADER contained was 338,224. The RORs (95% confidential interval, CI) of drospirenone combined with ethinyl estradiol (EE, Dro-EE), norethisterone with EE (Ne-EE), levonorgestrel with EE (Lev-EE), desogestrel with EE (Des-EE), and norgestrel with EE (Nor-EE) were 56.2 (44.3-71.4), 29.1 (23.5-35.9), 42.9 (32.3-57.0), 44.7 (32.7-61.1), and 38.6 (26.3-56.7), respectively. The medians (25%-75%) of the time-to-onset of Dro-EE, Ne-EE, Lev-EE, Des-EE, and Nor-EE were 150.0 (75.3-314.0), 128.0 (27.0-279.0), 204.0 (44.0-660.0), 142.0 (41.3-344.0), and 16.5 (8.8-32.0) days, respectively. The 95% CIs of the WSP-ß for Ne-EE, Lev-EE, and Nor-EE were lower and excluded 1. Association rule mining indicated that patients with anemia had a potential risk of developing a TE when using CEPs. Our results suggest that it is important to monitor patients administered CEP for TE. Careful observation is recommended, especially for those using Nor-EE, and this information may be useful for efficient therapeutic planning.

Endoscopic surgical management of sinonasal inverted papilloma extending to frontal sinuses.

OBJECTIVE: Sinonasal inverted papilloma has been traditionally managed with external surgical approaches. Advances in imaging guidance systems, surgical instrumentation, and intraoperative multi-visualization have led to a gradual shift from external approaches to endoscopic surgery. However, for anatomical and technical reasons, endoscopic surgery of sinonasal inverted papilloma extending to the frontal sinuses is still challenging. Here, we present our experience in endoscopic surgical management of sinonasal inverted papilloma extending to one or both frontal sinuses. METHODS: We present 10 cases of sinonasal inverted papilloma extending to the frontal sinuses and successfully removed by endoscopic median drainage (Draf III procedure) under endoscopic guidance without any additional external approach. RESULTS: The whole cavity of the frontal sinuses was easily inspected at the end of the surgical procedure. No early or late complications were observed. No recurrence was identified after an average follow-up period of 39.5 months. CONCLUSION: Use of an endoscopic median drainage approach to manage sinonasal inverted papilloma extending to one or both frontal sinuses is feasible and seems effective.

Effect of terfenadine and pentamidine on the HERG channel and its intracellular trafficking: combined analysis with automated voltage clamp and confocal microscopy.

The effects of terfenadine and pentamidine on the human ether-a-go-go related gene (hERG) channel current and its intracellular trafficking were evaluated. Green fluorescent protein (GFP)-linked hERG channels were expressed in HEK293 cells, and the membrane current was measured by an automated whole cell voltage clamp system. To evaluate drug effects on channel trafficking to the cell membrane, the fraction of channel present on the cell membrane was quantified by current measurement after drug washout and confocal microscopy. Terfenadine directly blocked the hERG channel current but had no effect on trafficking of hERG channels to the cell membrane after application in culture medium for 2 d. In contrast, pentamidine had no direct effect on the hERG channel current but reduced trafficking of hERG channels. The two drugs inhibited hERG channel function through different mechanisms: terfenadine through direct channel blockade and pentamidine through inhibition of channel trafficking to the cell membrane. Combined use of automated voltage clamp and confocal microscopic analyses would provide insights into the mechanisms of drug-induced QT-prolongation and arrhythmogenesis.

[Continuous hemodiafiltration during graft repair for a ruptured thoracoabdominal aortic aneurysm was useful in a patient with chronic renal failure].

A 74-year-old man with ruptured thoracoabdominal aortic aneurysm was scheduled for open surgical repair under partial cardiopulmonary bypass. He had a history of diabetes mellitus and a concomitant renal dysfunction, requiring regular intermittent hemodialysis. To maintain electrolytes, acid base as well as water balance within adequate ranges, we planned to use continuous hemodiafiltration (CHDF) during the surgery because there was a high incidence of bolus transfusion to deal with massive bleeding in these surgeries. We increased fluid removal speed of ultrafiltration when blood components had to be infused rapidly. With these considerations, the patient did not develop fluid overload, hyperkalemia, or aggravation of acidosis. We did not administer anticoagulants into CHDF circuit because activated coagulation time was prolonged probably due to massive bleeding prior to the surgery. Heparin was administered just before the partial car diopulmonary bypass. There was no evidence for thromboembolic complications due to CHDF use. In conclusion, we successfully managed electrolytes as well as acid base balance, and hydration of a patient with chronic renal failure by using CHDF during open graft repair of ruptured thoracoabdominal aortic aneurysm.

Analysis and comparison of clinical results of bilateral sagittal split ramus osteotomy performed with the use of monocortical locking plate fixation or bicortical screw fixation.

OBJECTIVE: The aim of this study was to compare clinical results of bilateral sagittal split ramus osteotomy (BSSRO) with the use of monocortical locking plate or bicortical screw fixation. STUDY DESIGN: Fifty-five patients underwent BSSRO for prognathism, using either monocortical locking plate (group A; n = 28) or bicortical screw (group B; n = 27) osseofixation. No intermaxillary fixation was done after surgery. Groups were subdivided according to presence or absence of mandibular asymmetry. Time course changes in condylar and skeletal stability were measured on lateral and posteroanterior cephalograms and axial radiographs before surgery and at 3 and 6 months after surgery. RESULTS: In facial symmetry subjects, the change in angle of the longitudinal axis of the condyle in group A was significantly greater than that for group B up to 3 months after surgery, but no significant differences were found in facial asymmetry subjects. CONCLUSIONS: The findings of this study suggest that monocortical fixation using the locking plate system to stabilize SSRO is as reliable as bicortical screw fixation regardless of facial asymmetry.

Association of lymphocyte crossmatch and the outcome of intestinal transplantation in swine.

BACKGROUND: The effect of preformed antidonor antibodies have been demonstrated in various types of solid organ transplantation. However, the significance of anti-donor antibodies in intestinal transplantation remains unclear. The aim of this study is to evaluate the impact that the extent of T cell crossmatch has on the outcome of swine intestinal transplantation. MATERIALS AND METHODS: All studies were performed on outbred domestic male pigs weighing from 15 to 20 kg. Intestinal transplantation was performed orthotopically with an exchange of grafts between white and black pigs. FK506 was administered intravenously (0.1 mg/kg per day, POD 0-7) for immunosuppression. A lymphocyte crossmatch test was performed using the direct CDC crossmatch. The results were considered positive when more than 10% of the donor lymphocytes were killed by the recipient's serum. In addition, 0-10, 11-20, 21-30, 31-80 and 81-100% of the killed lymphocytes were classified as grade 1, 2, 4, 6 and 8, respectively. RESULT: A total of 34 intestinal transplantations were performed. All but one case had positive donor specific T cell crossmatches. The number of grade 2, 4, 6 and 8 cases was 11, 14, 6 and 2, respectively. Although there was a tendency towards a decreased survival according to the grade, the survival rate was not statistically different among each different grade. Moreover, the rates of acute cellular rejection and vascular complications were not significantly different among the four grades. CONCLUSION: These results suggest that the extent of positive T cell crossmatch is not associated with the outcome of swine intestinal transplantation.

Umbilical crease incision for duodenal atresia achieves excellent cosmetic results.

BACKGROUND: The surgical procedure for treating congenital duodenal atresia has normally been performed by an upper abdominal transverse incision. Recently, duodenoduodenostomy has been attempted using an umbilical crease incision to improve the cosmetic results. METHODS: Eighteen cases of duodenal obstruction, including 15 atresia, 2 stenosis, and 1 atresia and stenosis, were treated from June 2001 to August 2009, in which 8 cases were performed via the umbilical crease incision and 10 cases via the conventional transverse incision. The clinical records of all cases were evaluated retrospectively. RESULTS: All cases underwent radical operation safely. There were no differences in the operating time between the two kinds of incision. Two cases of umbilical crease incision showed minor complications. All the cases operated via the umbilical crease incision achieved a scarless abdomen within a few months after the operation. CONCLUSIONS: The outcome of duodenal atresia is satisfactory with excellent cosmesis after a duodenoduodenostomy performed via the umbilical crease incision.

Excellent long-term outcome of hepaticojejunostomy for biliary atresia with a hilar cyst.

BACKGROUND: Biliary atresia (BA) with hilar cyst is an uncommon variant, which constitutes less than 10% of all types of BA, and the operative procedure for this type of BA remains controversial. METHODS: We have had 200 cases with BA from 1963 to 2008 in our institute and our branch hospitals, in which 12 cases (6%) were BA with a hilar cyst. The clinical records of all cases were evaluated retrospectively. RESULTS: Twelve BA patients with a hilar cyst included 2 boys and 10 girls. The diagnosis of BA was confirmed by intraoperative cholangiography (cloudy or treelike pattern). In all 12 cases, a hepaticojejunostomy was performed at a median age of 71.6 days (range, 24-136 days). The follow-up periods were 1.2 to 23.2 years. The current mean total bilirubin level was 0.8 mg/dL (range, 0.2-3.5 mg/dL), and the mean direct bilirubin level was 0.2 mg/dL (range, 0.0 to 0.8 mg/dL). Methylprednisolone or oral prednisolone was administered in 8 cases after operation, and 10 of 12 cases achieved a jaundice free state. The postoperative complications were cholangitis (n = 10), gastroesophageal varices (n = 7), splenomegaly (n = 3), ileus (n = 1), and pulmonary hypertension (n = 1). The overall survival rate with a native liver was 10 (83.3%) of 12 cases, of which 9 cases were jaundice-free and only 1 case showed recurrent jaundice. The other 2 cases underwent living-donor liver transplantation at age of 2 and 20 years, respectively, and they are currently doing well. CONCLUSIONS: Most cases of BA with a hilar cyst achieved excellent clinical outcome after a hepaticojejunostomy.

Surgical complications after living donor liver transplantation in patients with biliary atresia: a relatively high incidence of portal vein complications.

BACKGROUND/PURPOSE: The aim of this study is to present the surgical complications in living donor liver transplantation (LDLT) for biliary atresia (BA) as a treatment for end stage liver disease. PATIENTS AND METHODS: Twenty-nine LDLTs were performed in patients with BA between October 1996 and April 2008 in Department of Pediatric Surgery at Kyushu University Hospital. The initial immunosuppression was a combination of tacrolimus and steroids. RESULTS: Twenty-eight of 29 cases with BA, who previously underwent Kasai's operation and LDLT was performed at a median age of 9.1 years (range 7 months to 28 years). Only one case was performed primary LDLT. Post-transplant complications included portal vein complications (n = 5), three of which successfully treated by Rex-shunt or ballooning. Others were bile leakage (n = 4), intestinal perforation (n = 4), and so on. The overall survival rate was 86.2% (25/29). One patient died of chronic rejection, surgical complications after LDLT in BA while others died of sepsis, multi-organ failure, and brain hemorrhage. CONCLUSION: The incidence of portal vein complications and intestinal perforations was relatively high in LDLT for BA, possibly due to inflammation of the hepatoduodenal ligament and colonic adhesion to the liver. It is important to make an accurate diagnosis at an early stage and provide appropriate treatment.

Successful endoscopic unroofing for a pediatric choledochocele.

A choledochocele (type 3 choledochal cyst) is rarely observed in children. A 6-year-old boy experienced abdominal pain approximately 6 times in the previous 5 months. Magnetic resonance cholangiopancreatography findings revealed a small cyst in the distal common bile duct. An endoscopic examination was performed, and a choledochocele was detected at the duodenal papilla. Endoscopic unroofing was successfully performed, and the postoperative course was uneventful.

Periventricular echodensity measured with the integrated backscatter system: from a qualitative assessment to a quantitative approach.

BACKGROUND: The degree of periventricular white matter echodensity in preterm infants has been utilized as a sign of the early ultrasonographic appearance of periventricular leukomalacia, and this has been called periventricular echodensity (PVE). OBJECTIVES: The aim of this study was to quantitatively measure PVE utilizing a new method which is called calibrated integrated backscatter (calibrated IB). METHODS: Eighty-eight preterm infants (extremely low birth weight infants, n = 17; very low birth weight infants, n = 26; low birth weight infants, n = 45) without any CNS abnormality were enrolled. IB is the returned sound pressure against supersonic waves sent from an ultrasonographic machine. The IB of the choroid plexus and periventricular white matter in the subrolandic area were measured on a parasagittal cerebral image. The degree of PVE was defined by subtracting the IB of the choroid plexus from that of the periventricular white matter in the subrolandic area (calibrated IB of PVE). RESULTS: The intraobserver and interobserver correlations were both excellent (between 0.87 and 0.98 as correlation coefficients). There was a trend for the calibrated IB of PVE to decrease in accordance with time after birth, with a significant difference in very low birth weight and low birth weight infants. CONCLUSIONS: The objectively measured brightness of PVE was comparable to that of the choroid plexus irrespective of the size of the infants. Measurement of the calibrated IB of PVE might be a reliable method to assess PVE.

Subcutaneous emphysema, pneumomediastinum, pneumothorax, pneumoperitoneum, and pneumoretroperitoneum by insufflation of compressed air at the external genitalia in a child.

A 7-year-old girl with concurrent subcutaneous emphysema, pneumomediastinum, pneumothorax, pneumoperitoneum, and pneumoretroperitoneum arrived at our facility. Compressed air at 5 atm of pressure was insufflated through the nozzle of a spray gun over her external genitalia. She was admitted for a small amount of genital bleeding and significant subcutaneous emphysema extending from the cheek to the upper body. Radiographic examination of the abdomen was suggestive of a visceral perforation, but she was managed conservatively and discharged in satisfactory condition without surgical intervention. The female genitalia possibly served as the entry point for air into the retroperitoneum and peritoneal cavity, with subsequent migration of air through the esophageal hiatus that resulted in pneumomediastinum, pneumothorax, and extensive subcutaneous emphysema.

Diagnostic value of lectin reactive alpha-fetoprotein for neoinfantile hepatic tumors and malignant germ cell tumors: preliminary study.

BACKGROUND AND PURPOSE: The serum alpha-fetoprotein (AFP) level has been used as a tumor marker for hepatoblastoma, and malignant germ cell tumors in pediatric patients. The AFP has 3 isoforms (L1, L2, L3), and the usefulness of the L3 fraction as a diagnostic marker for the adult hepatocellular carcinoma is well known. However, there are few reports dealing with various pediatric malignant tumors. In the current study, we analyzed the diagnostic value of AFP fractions for pediatric diseases, in particular, those occurring in the neoinfantile period. MATERIALS AND METHODS: From 2003 to 2006, two cases of hepatoblastoma, and 5 cases of germ cell tumor, all of which were neoinfantile, were treated in our department. In our analytical system (LiBASys), the level of the L3 fraction contains the majority of the L2 fraction. The total AFP (ng/mL) level and the L3 fraction (%) were measured to assess the usefulness of the L3 fraction as a diagnostic marker. RESULTS: In all cases of hepatoblastoma and yolk sac tumor, both the total AFP and the L3 fraction were high, either before treatment or in the presence of malignant tumors. Most of the cases of neonatal immature teratoma showed a high total AFP level during the neoinfantile period, however, the L3 fraction was around 10%, and decreased after surgical treatment. Only 1 case of the immature teratoma demonstrated malignant transformation, when the patient was 8 months old. As the total AFP and the AFP-L3 fraction were proportionally elevated, the patient was treated with additional surgical resection and chemotherapy. In the case of neonatal mature teratoma, the L3 fraction was below 0.5%, even when the total AFP level was high. DISCUSSION: Our results indicated that the level of the L3 fraction accurately confirmed the existence, or the malignant potential of hepatic tumor or germ cell tumor. The L3 fraction is useful as a tumor marker during the neoinfantile period.

Alterations of RB1 gene in embryonal and alveolar rhabdomyosarcoma: special reference to utility of pRB immunoreactivity in differential diagnosis of rhabdomyosarcoma subtype.

PURPOSE: Rhabdomyosarcoma (RMS), which is the most common pediatric soft tissue sarcoma, is classified into two major histologic subtypes, embryonal RMS (ERMS) and alveolar RMS (ARMS). RMS is occasionally reported to be the second neoplasm of hereditary retinoblastoma. Osteosarcoma is known as the most common second neoplasm of hereditary retinoblastoma, and tumorigenesis of osteosarcoma has been proven in previous studies to be related to the RB gene (RB1) alteration. Therefore, there might be a correlation between the tumorigenesis of RMS and RB1 alteration. METHODS: We examined the RB protein (pRB) expression and RB1 alteration such as allelic imbalance (gain or loss) and homozygous deletion, using immunohistochemistry, microsatellite makers, and quantitative real-time PCR in 57 sporadic RMS. RESULTS: Allelic imbalance was more frequently detected in ERMS (13/27), than in ARMS (3/20) (P = 0.04). Homozygous deletion on the protein-binding pocket domain of RB1 was found in 6 of 27 ERMS and in 2 of 20 ARMS (P = 0.24). Furthermore, immunohistochemical pRB labeling indexes (LI) in 31 ERMS (median value, 31%) were significantly reduced in comparison with those observed in 26 ARMS (median value, 85%) (P < 0.0001). CONCLUSIONS: Our results support the assertion that tumorigenesis of RMS may be associated with RB1 alteration especially in ERMS, as previously reported for osteosarcoma. As for the RB pathway, each subtype of RMS may have a different tumorigenesis. In addition, immunohistochemical pRB LI may have the potential to be a useful ancillary tool in the differential diagnosis of RMS subtypes.