Novel device prototyping for endoscopic cell sheet transplantation using a three-dimensional printed simulator.

INTRODUCTION: Considering higher risks of candidates for cardiac regenerative therapy with compromised cardiac function, it is anticipated to develop less invasive surgical procedures. In the present study, we aimed to develop a prototype of totally endoscopic cell sheet delivery device and evaluate the surgical technique for epicardial cell sheet placement using three-dimensional (3D) printed simulators based on human computed tomography data. METHODS: We designed an endoscopic cell sheet delivery device with outer and inner frame with self-expandable applicator which can be opened in thoracic cavity. We launched spout line to provide liquids on the applicator surface and tension line to gently bend the applicator dorsally. We prepared human mesenchymal stem cell (MSC) sheets and compared wet/dry conditions of 3D printed heart/porcine heart and applicator to identify suitable conditions for cell sheet transplantation. Finally we validated the feasibility of endoscopic transplantation to anterior and lateral wall of left ventricle using 3D printed simulators. RESULTS: Moist condition of both 3D printed heart/porcine heart surface and applicator at transplantation yielded highest successful rate (100%, p = 0.0197). For both endoscopic transplantation sites, MSC sheets were successfully deployed. The procedure duration was 157 ± 23 s for anterior wall and 123 ± 13 s for the lateral wall in average, respectively. CONCLUSIONS: We developed a novel prototype of endoscopic cell sheet delivery device for minimally-invasive cardiac regenerative therapy utilizing a 3D printed simulator. The commercialization of the prototype may provide a safe minimally-invasive method to deliver potential cardiac regenerative therapy in the future.

Cytoplasmic maspin expression predicts poor prognosis of patients with soft tissue sarcomas.

BACKGROUND: Maspin is a 42 kDa protein known to act as a tumor suppressor. Although its function has not been fully elucidated, numerous reports have investigated the prognostic impact of maspin in patients with several types of cancer. However, there have been no reports on the association between maspin expression and the prognosis of patients with soft tissue sarcomas (STS). The aim of this study was thus to explore the association of maspin expression with the prognosis of patients with STS. METHODS: One-hundred and eight paraffin-embedded STS tissue samples were immunohistochemically analyzed using antibodies for maspin and Ki-67 antigen. The patients were followed up for 1 to 300 months (median: 33 months) and the prognostic value was evaluated by log-rank test and Cox's regression hazard model. RESULTS: Cytoplasmic maspin expression was observed in 48.1% of specimens, and was significantly correlated with a higher FNCLCC grade (P = 0.002) and the presence of distant metastases (P = 0.001), and those with cytoplasmic maspin expression had both shorter disease-free survival (DFS) and overall survival (OS) by log-rank test (P <0.001, P = 0.001, respectively). By Cox's multivariate analysis, the presence of distant metastases was the only prognostic factor for DFS and OS. CONCLUSIONS: This is the first report to reveal an association between maspin expression and the prognosis of patients with STS. Although further studies with a larger series of patients and a longer follow-up period will be needed, cytoplasmic maspin expression could be an indicator of unfavorable prognosis in patients with STS. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_205.

Sacral chordoma in an adult showing an aggressive clinical course: A case report.

The current report presents a case of a 78-year-old male with sacral chordoma, showing an aggressive clinical course. The patient underwent sacral resection, however, nine months later, multiple metastases were detected by magnetic resonance imaging. The metastases progressed rapidly and 15 months following surgery the patient succumbed to respiratory dysfunction. An autopsy revealed multiple metastases of the lung, liver, heart, kidneys and vertebrae. Pathologically, the tumors did not show proliferation of anaplastic cells or dedifferentiation; however, the metastatic tumor cells were smaller than the primary tumor cells. The Ki-67 labeling indices were <5% in all of the patient's tumors, therefore, the capacity for cellular proliferation of the tumors was considered to be low. Chordoma in adults are generally slow-growing tumors and are associated with a relatively prolonged course and frequent local recurrences. Therefore, it must be recognized that chordoma may grow rapidly and show an aggressive clinical course, even when the Ki-67 labeling index is low.

Proliferative fasciitis mimicking a sarcoma in a child: a case report.

Proliferative fasciitis (PF) is a benign, discrete proliferation of fibroblasts or myofibroblasts in soft tissue. Proliferative fasciitis mostly occurs in adults and is often confused with a sarcoma because of its rapid growth and peculiar histological features. We report a case of PF mimicking a sarcoma which developed in a 13-year-old boy, who noticed a painful tumor, with gradual enlargement, in his right lower leg. Magnetic resonance imaging revealed that the tumor measured 34 mm × 20 mm × 41 mm and was located in the subcutaneous tissue. The tumor was surgically resected. Pathologically, the tumor was composed of a proliferation of atypical spindle cells, admixed with larger ganglion-like cells. Immunohistochemically, the tumor cells were positive for vimentin, cytokeratin, smooth muscle actin, HHF-35 and Fli-1. The tumor was subsequently diagnosed as a PF, although it was difficult to differentiate from a sarcoma. Five years after surgery, the postoperative course has been uneventful with no recurrence or metastasis.

Hemoglobin maintenance and dosing strategies using intravenous continuous erythropoietin receptor activator in Japanese hemodialysis patients.

Methoxy polyethylene glycol-epoetin beta, a continuous erythropoietin receptor activator (CERA), is reported to be effective in managing renal anemia but there is little data about CERA in Japan. This study aimed to ascertain the effects of CERA in Japanese hemodialysis patients and the appropriate starting dose of CERA when switching from other erythropoiesis-stimulating agents. We switched 61 stable hemodialysis patients to 4-weekly intravenous CERA, from either epoetin beta (rHuEPO) or darbepoetin alpha (DA). When determining the initial dose of CERA, we used guidelines recommended by the Japanese supplier for switching from rHuEPO, but for DA we based the CERA dose on European reports, because no Japanese guidelines exist. Fifty-two patients completed the 28-week study. Hemoglobin was maintained within the target range (10.0-12.0 g/dL). The required CERA dose decreased over the 28 weeks. The hemoglobin level and CERA dose stabilized faster when switching from DA. CERA showed similar efficacy in diabetic and non-diabetic patients. The effect of CERA is similar regardless of whether patients switch from low- or high-dose erythropoiesis-stimulating agents. In conclusion, CERA is effective for Japanese hemodialysis patients at a lower dose than expected.

Intramuscular myxoma of the buttock mimicking low-grade fibromyxoid sarcoma: diagnostic usefulness of MUC4 expression.

Intramuscular myxoma (IM) is a benign intramuscular neoplasm composed of fibroblasts and abundant myxoid stroma. Some malignant soft tissue tumors can undergo myxomatous degeneration, which makes it difficult to distinguish them from IM. We describe a case of IM of the buttock region mimicking low-grade fibromyxoid sarcoma. The tumor appeared as a well-defined ovoid mass with a cystic lesion on MRI images, and mild uptake on PET images was seen. This was originally misdiagnosed as low-grade fibromyxoid sarcoma (LGFMS) after core-needle biopsy. The mass was excised en bloc and sent for histology. The surgical specimen showed the features of LGFMS with the same characteristics as those mentioned in the previous biopsy report. After surgery, MUC4 expression, a highly sensitive and specific immunohistochemical marker for LGFMS, and FUS gene rearrangement by FISH was not detected upon re-examination; therefore, a conclusive diagnosis of IM was made. The patient had no local recurrence at the 3-year follow-up. Our case suggests that IM with mild FDG uptake is frequently confused with other low-grade malignant myxoid tumors. In addition, absence of MUC4 expression is the definitive key to distinguish IM from LGFMS.

Multifocal localized pigmented villonodular synovitis arising separately from intra- and extra-articular knee joint: case report and literature review.

We present a case of intra- and extra-articular localized pigmented villonodular synovitis (PVNS) of the right knee joint separately. A 14-year-old girl presented with a painful mass over the medial aspect of the right knee. MRI demonstrated nodular masses located in the anteromedial area of the knee joint and on the surface of the deep medial collateral ligament outside the joint. Arthroscopic and open synovectomy were performed for both lesions. Histopathology confirmed diagnosis of localized PVNS and pigmented villonodular bursitis, respectively. Our case highlights that localized PVNS are likely to occur in intra- and extra-articular regions separately.

Angiosarcoma of the proximal humerus: a case report and review of the literature.

INTRODUCTION: Angiosarcoma of bone is an uncommon primary bone neoplasm that is composed of tumor cells that show endothelial differentiation. This is an aggressive malignancy characterized by frequent local recurrence and distant metastases. The majority of patients die within one year of diagnosis, and this shows that angiosarcoma of bone is an aggressive high-grade tumor. CASE PRESENTATION: We present the case of a 65-year-old Japanese woman who had primary angiosarcoma of the proximal humerus with a pathological fracture. An open biopsy confirmed a diagnosis of primary angiosarcoma of bone. Our patient was treated with neoadjuvant chemotherapy and wide resection. One month after surgery, she developed multifocal distant metastasis to her liver and spleen. CONCLUSIONS: Angiosarcoma of the humerus is extremely rare. Radiographically, there is no specific finding associated with angiosarcoma of bone as opposed to other malignant bone tumors. The cornerstone of treatment is en bloc resection followed by as much adjuvant radiation therapy as possible. However, the role of chemotherapy remains undefined, and better systemic agents are clearly needed.

Clinical features and surgical outcomes of cervical myelopathy in the elderly.

UNLABELLED: Treatment of cervical myelopathy in elderly patients is controversial. We retrospectively studied 113 patients who had decompression surgery from 1990-2001 to clarify how pre-operative conditions, duration of symptoms, involved levels, surgical outcomes, and complications differ between younger and elderly patients. We also asked whether elderly patients would likely have reasonable outcomes of surgery. The patients were divided into five age groups: Group 1, 36-45 years (12 patients); Group 2, 46-55 years (22 patients); Group 3, 56-65 years (31 patients); Group 4, 66-75 years (32 patients); and Group 5, 76-85 years (16 patients). The duration of symptoms was similar among the five groups. The involved level was more cephalic in the older groups, and the most frequently involved level in patients older than 75 years was C3-C4. Neurologic symptoms were more severe preoperatively and postoperatively in older patients. The recovery ratio also was lower in older patients; however, in Group 5 it was 36.9%, which indicated that patients older than 75 years could regain approximately 40% of their function postoperatively. Decompression surgery can be a reasonable treatment option for cervical myelopathy, even in elderly patients. LEVEL OF EVIDENCE: Therapeutic Study, Level IV (case series). See the Guidelines for Authors for a complete description of levels of evidence.

Cyclooxygenase-2 in human malignant fibrous histiocytoma: correlations with intratumoral microvessel density, expression of vascular endothelial growth factor and thymidine phosphorylase.

Cyclooxygenase-2 (COX-2) has been found to be up-regulated in several types of human malignant tumors and proposed to have a role in the angiogenic process. This study examined the expression of COX-2 in two human malignant fibrous histiocytoma (MFH) cell lines by Western blotting, which showed a specific single band at 72 kDa. Immunohistochemistry was conducted in 35 MFHs and 30 benign fibrohistiocytic tumors (BFHTs), comparing the expression of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP), as well as intratumoral microvessel density (IMVD). COX-2 expression was noted in 22 (62.9%) MFHs, but in no BFHT. IMVD values were significantly higher in the MFHs (90.6+/-8.0) than BFHTs (27.9+/-3.1), and also in the COX-2 positive (104.5+/-11.3) than negative (67.2+/-5.8) MFHs. VEGF and TP expression was also associated with a significantly higher level of COX-2, as well as greater IMVD. The highest IMVD values were noted in the 17 MFHs (120.8+/-11.5) expressing all three factors. Clinical analysis demonstrated poorer survival in the 18 COX-2 positive MFHs than in the 10 negative ones, although the small number of cases did not reveal a significant difference. The results overall indicated that COX-2 expression is associated with intratumoral angiogenesis, which might provide favorable conditions for tumor progression in human MFHs.

Inhaled nitric oxide attenuates apoptosis in ischemia-reperfusion injury of the rabbit lung.

BACKGROUND: Lung ischemia-reperfusion injury occurs after lung transplantation and various clinical procedures. Recently, apoptosis was reported to be induced after ischemia-reperfusion. We investigated the effects of inhaled nitric oxide (NO) on lung ischemia-reperfusion and apoptosis after ischemia-reperfusion. METHODS: As a control group, the left pulmonary hilum of Japanese white rabbits (n = 10) was occluded for 120 minutes and reperfused for 120 minutes. In the inhaled NO group (n = 10), 20 parts per million nitric oxide was inhaled during reperfusion. The sham-operated group was ligated at the right hilum and perfused by the left lung only for 120 minutes. The mean pulmonary arterial pressures and Pao2 were measured during reperfusion. The wet-to-dry weight ratio of the left lower lobe of the lung was calculated. The number of apoptotic cells was estimated using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) technique. The TUNEL staining for a time course study was done using 15 control animals that were killed by exsanguination at 15, 30, and 60 minutes after reperfusion. RESULTS: After 120 minutes of reperfusion, the mean pulmonary arterial pressures in the control group and in the inhaled NO group were 23.0 +/- 3.2 mm Hg and 13.6 +/- 2.4 mm Hg, respectively (p < 0.01). At the same time point, the Pao2 in the control group and in the inhaled NO group were 46.1 +/- 15.9 mm Hg and 88.1 +/- 14.7 mm Hg, respectively (p < 0.01). The wet-to-dry weight ratios in the control group and in the inhaled NO group were 0.856 +/- 0.024 and 0.808 +/- 0.006, respectively (p < 0.01). Apoptotic cells appeared in the early phase of reperfusion (after 15 minutes' reperfusion). The number of apoptotic cells was significantly lower in the inhaled group than in the control group after 120 minutes' reperfusion (1.76% versus 2.87%, p < 0.01). CONCLUSIONS: Our results suggest that the inhaled NO prevents lung ischemia-reperfusion injury and attenuates apoptosis after reperfusion in the rabbit lung.

Preoperative irradiation combined with chemotherapy impairs healing of bronchial anastomosis during the early postoperative period in rats.

The effect of preoperative irradiation and antineoplastic agents on healing at the site of bronchial anastomosis was investigated using rats. The bursting pressure in irradiation group and combined irradiation and chemotherapy group was significantly lower than in control and chemotherapy group at day 5 after operation. There was no significant difference in bursting pressure in all groups at day 7. The histologic finding of the anastomosis with H & E stain showed that submucosal connective tissue had not regenerated, and defects were seen in the submucosal tissue in irradiation and combined therapy group at day 3 and day 5. But, the connective tissue had matured in irradiation group at day 7 compared with control group. In conclusion, this study demonstrated that the healing of bronchial anastomosis was markedly delayed in early postoperative days in the rats receiving irradiation and combined therapy.

A selective cyclooxygenase-2 inhibitor, NS-398, inhibits cell growth by cell cycle arrest in a human malignant fibrous histiocytoma cell line.

BACKGROUND: NS-398, a selective cyclooxygenase-2 (COX-2) inhibitor, has been shown to suppress cell proliferation and induce apoptosis in a variety of human cancer cell lines in vitro, except for sarcoma cell lines. The aim of this study was to examine the effect of NS-398 on two human malignant fibrous histiocytoma (MFH) cell lines: MFH-ino and MFH-ToE. MATERIALS AND METHODS: We investigated the effect of NS-398 at various concentrations on the growth of MFH cell lines using cell proliferation assay, cell cycle analysis and EIA assay of PGE2 production. RESULTS: Western blot analysis revealed COX-2 protein expression in both MFH cell lines. NS-398 at 10 or 100 microM resulted in inhibition of the cell proliferation in a dose- and time-dependent manner. NS-398 at 100 microM also induced G0/G1 arrest accompanied by up-regulation of P21WAF1 in MFH-ino cells in a time-dependent manner until 72 hours. NS-398 slightly increased the number of cells in the G2/M-phase and decreased the number in the G0/G1-phase in MFH-ToE cells lacking p53 gene function. Moreover, NS-398 down-regulated PGE2 production in the MFH-ToE cells in a time-dependent manner, but not in the MFH-ino cells. CONCLUSION: Our results indicate that NS-398 inhibits the cell growth and induces G0/G1 arrest in MFH-ino cells accompanied with up-regulation of P21WAF1.

Mast cells in human allografted kidney: correlation with interstitial fibrosis.

INTRODUCTION: Chronic allograft nephropathy is the major cause of long-term graft failure in human allografted kidney transplantation. In addition to macrophages and T lymphocytes, mast cells have been shown to increase in chronic allograft nephropathy. The present study examined tryptase-positive mast cells and microvessels in the allografted kidney. MATERIALS AND METHODS: We selected 131 biopsy specimens obtained from 100 allografted, 14 non-grafted renal biopsy specimens and nine nephrectomy specimens due to renal cell carcinomas. Formalin-fixed, paraffin- embedded specimens were immunostained using primary antibodies for mast cell tryptase, mast cell chymase and CD34. The number of the mast cells and microvessels was counted in at least 20 high-power fields (10 x 40). RESULTS: Tryptase-positive mast cells outnumbered chymase, toluidine blue or naphthol-AS-D choloacetate-positive mast cells. The mean number of the tryptase-positive mast cells was significantly higher in the 36 specimens with chronic allograft nephropathy (5.1 +/- 3.5) among the grafted kidneys with other disease categories (P < 0.001). In the chronic allograft nephropathy, the mean numbers of mast cells was significantly higher in Ci 2 + Ci 3 (n = 20; 6.4 +/- 3.89) than in Ci 1 (n = 16; 3.6 +/- 2.65) (P < 0.01). In the non-grafted kidney, the number of mast cells was highest in the four specimens with diabetic nephropathy. Mast cells and microvessels were analysed in the two representative cases, which subsequently developed chronic allograft nephropathy. Both of the cases showed the highest number of mast cells in chronic allograft nephropathy. In contrast, the mean number of microvessels tended to decrease along with interstitial fibrosis. CONCLUSIONS: This study demonstrated clearly a close association between renal interstitial fibrosis and mast cells, which might play an important role in the development of chronic allograft nephropathy.

[CPT-11 hepatic arterial injection plus oral UFT administration for liver metastasis of rectal cancer--report of two cases].

The first patient was a 51-year-old male who had 5-fluorouracil-resistant recurrent rectal cancer with multiple liver metastases. He was given our new combination chemotherapy consisting of hepatic arterial injection of CPT-11 (20 mg/body) on day 1 and day 2 and oral administration of UFT (300 mg/day) on days 3 to 6 of a 7 day cycle starting in January 2001. Six weeks after the beginning of chemotherapy, the liver metastatic lesions were reduced. He is now living with outpatient treatment. The second patient was a 76-year-old male who had initial recurrent rectal cancer with multiple liver metastases. Thirty-two weeks after the same chemotherapy, the metastatic lesions had completely disappeared. Twelve months have passed since this chemotherapy, and we have not found any recurrent tumor. While significant antitumor effects were observed, there were few adverse events in either patient. These results suggest that combined chemotherapy of CPT-11 by hepatic arterial injection and oral administration of UFT is an effective treatment for liver metastases of rectal cancer.

Expression of minichromosome maintenance-2 in human malignant fibrous histiocytomas: Correlations with Ki-67 and P53 expression, and apoptosis.

This study examined the clinicopathological significance of minichromosome maintenance-2 (MCM2) expression in 38 human malignant fibrous histiocytomas (MFHs) and 36 benign fibrohistiocytic tumors (BFHTs) immunohistochemically, and in 9 human sarcoma or carcinoma cell lines, as well as 7 surgical specimens by Western blotting. MCM2 was detected in all the cell lines and surgical specimens as a single band at 120 kDa, while P53 expression was variable. Nuclear expression of MCM2 was noted in tumor but not mitotic cells of all the MFHs and 26 (72.2%) of the BFHTs, the labeling indices (LIs) being 62.0% in the 28 ordinary types, 38.5% in the 10 myxoid types, and 11.2% in the BFHTs with significant difference. Moreover, the LI was significantly higher for MCM2 than that for Ki-67 in the MFHs of both types (p<0.05). No correlation was noted between the MCM2-LI and P53 expression or apoptotic indices, which were significantly higher in the MFHs than BFHTs (p<0.01). These results indicate that MCM2 would correlate with cell proliferation rather than apoptosis in MFHs, and the expression is ubiquitous in proliferating cells, regardless of the expression of P53. Thus, MCM2 might be a reliable marker of proliferating cells in human MFH.