Evaluation of a decellularized bronchial patch transplant in a porcine model.

Biological scaffolds for airway reconstruction are an important clinical need and have been extensively investigated experimentally and clinically, but without uniform success. In this study, we evaluated the use of a decellularized bronchus graft for airway reconstruction. Decellularized left bronchi were procured from decellularized porcine lungs and utilized as grafts for airway patch transplantation. A tracheal window was created and the decellularized bronchus was transplanted into the defect in a porcine model. Animals were euthanized at 7 days, 1 month, and 2 months post-operatively. Histological analysis, immunohistochemistry, scanning electron microscopy, and strength tests were conducted in order to evaluate epithelialization, inflammation, and physical strength of the graft. All pigs recovered from general anesthesia and survived without airway obstruction until the planned euthanasia timepoint. Histological and electron microscopy analyses revealed that the decellularized bronchus graft was well integrated with native tissue and covered by an epithelial layer at 1 month. Immunostaining of the decellularized bronchus graft was positive for CD31 and no difference was observed with immune markers (CD3, CD11b, myeloperoxidase) at two months. Although not significant, tensile strength was decreased after one month, but recovered by two months. Decellularized bronchial grafts show promising results for airway patch reconstruction in a porcine model. Revascularization and re-epithelialization were observed and the immunological reaction was comparable with the autografts. This approach is clinically relevant and could potentially be utilized for future applications for tracheal replacement.

Single-center evaluation of prognostic factors for thymoma treated by surgery: a retrospective study.

BACKGROUND: This study aimed to retrospectively evaluate the clinical, pathological, and treatment-related factors associated with survival in patients with surgically treated thymomas. METHODS: Sixty patients with thymomas who underwent treatment at our institution between 2004 and 2015 were included. Survival analysis was performed based on curves that were obtained using the Kaplan-Meier method. The Wilcoxon test was used for all comparisons, and p < 0.05 was considered statistically significant. RESULTS: Forty-seven, four, three, four, and two patients presented tumor stages I, II, III, IVa, and IVb (according to the Masaoka classification), respectively, while six, 14, 11, 22, and seven patients had type A, AB, B1, B2, and B3 thymomas, respectively. Furthermore, 53 and eight patients underwent complete resection and required additional resection of adjacent organs, respectively, and no patients died from surgery-related complications. The five-year survival and recurrence-free survival (RFS) rates were 96 and 86%, respectively. The five-year survival rate for all stages was 100% except for those with stage IVb tumors (Masaoka classification); the survival rate for those patients was 0%. Separately, the five-year RFS rates for tumor stages I, II, III, IVa, and IVb were 100, 91, 91, 81, and 71%, respectively. Finally, the five-year survival rates in cases with complete and incomplete resections were 100 and 71%, respectively, indicating that the latter group had a significantly poorer prognosis (p < 0.001). CONCLUSIONS: These findings suggest that complete resection and the Masaoka pathological stage are significant predictors of prognosis in patients with thymomas. Surgery should aim to achieve complete resection; however, advanced cases may require multimodality therapy.

Pulmonary Metabolism of Substrates for Key Drug-Metabolizing Enzymes by Human Alveolar Type II Cells, Human and Rat Lung Microsomes, and the Isolated Perfused Rat Lung Model.

Significant pulmonary metabolism of inhaled drugs could have drug safety implications or influence pharmacological effectiveness. To study this in vitro, lung microsomes or S9 are often employed. Here, we have determined if rat and human lung microsomes are fit for purpose or whether it is better to use specific cells where drug-metabolizing enzymes are concentrated, such as alveolar type II (ATII) cells. Activities for major hepatic and pulmonary human drug-metabolizing enzymes are assessed and the data contextualized towards an in vivo setting using an ex vivo isolated perfused rat lung model. Very low rates of metabolism are observed in incubations with human ATII cells when compared to isolated hepatocytes and fewer of the substrates are found to be metabolized when compared to human lung microsomal incubations. Reactions selective for flavin-containing monooxygenases (FMOs), CYP1B1, CYP2C9, CYP2J2, and CYP3A4 all show significant rates in human lung microsomal incubations, but all activities are higher when rat lung microsomes are used. The work also demonstrates that a lung microsomal intrinsic clearance value towards the lower limit of detection for this parameter (3 µL/min/mg protein) results in a very low level of pulmonary metabolic clearance during the absorption period, for a drug dosed into the lung in vivo.

A case of massive hemothorax associated with an intrathoracic bronchogenic cyst.

A 34-year-old man presented with sudden back pain and dyspnea. Chest X-ray showed left-sided massive pleural effusion. Chest computed tomography revealed an intrathoracic mass sized 9 cm. Hemorrhagic effusion was achieved with thoracic drainage on admission. Diagnostic video-assisted surgery was indicated, and an unexpected cyst with bloody content was observed. The cyst was bluntly dissected from the pleura and removed from the diaphragm. The patient discharged uneventfully and there were no significant postoperative complications including bleeding or pneumothorax. Pathological observation of the cyst revealed pseudostratified ciliated epithelial cells, mucinous glands, and cartilage compatible with the diagnostic criteria for a bronchogenic cyst. Malignant transformation was not observed. Common clinical presentations of bronchogenic cysts include pain, dyspnea, and cough. Although rare, the risk of hemorrhage from bronchogenic cysts and subsequent development of hemothorax should not be underestimated.

Bronchial angiolipoma successfully treated by sleeve resection of the right bronchus intermedius: a case report.

BACKGROUND: Angiolipoma is a rare, benign tumor that primarily develops in the limbs and trunk. The occurrence of angiolipoma in the lungs is extremely rare; to date, only two cases of primary bronchial angiolipoma have been reported. Here, we report a case of angiolipoma of the right bronchus intermedius that was successfully treated with sleeve resection and reconstructive surgery. CASE PRESENTATION: This report presents a case of angiolipoma that developed in the right bronchus intermedius of a 68-year-old man. A chest CT revealed a 10-mm endobronchial mass that was clearly visible as a high-attenuation area of contrast enhancement. Bronchoscopy revealed a submucosal tumor on the anterior wall of the entrance to the right bronchus intermedius that was constricting the airway lumen. The tumor surface was covered with numerous engorged blood vessels, and the middle and inferior pulmonary lobes were intact. Bronchial sleeve resection of the right bronchus intermedius was performed. Histologically, a mixture of proliferating blood vessels and adipocytes were observed within the bronchus wall. Therefore, the pathological diagnosis was angiolipoma. Lung function was preserved, and complete resection of the tumor was achieved. At present (2 years and 7 months after surgery), the patient is recurrence-free. CONCLUSION: Accordingly, using bronchial sleeve resection and end-to-end anastomosis techniques, we accomplished complete tumor excision and avoided the need to resect additional lung parenchyma. Our procedure preserved pulmonary function and yielded a curative result. Bronchoscopic intervention or minimal parenchymal resection should be considered as treatments for bronchial angiolipoma. Given the small number of reports of bronchial angiolipoma, the collection of additional data is important to elucidate the clinical characteristics of this rare tumor.

Intrathoracic fibrolipoma resected using complete thoracoscopic surgery: a case report.

BACKGROUND: Other than adipocytes, lipomas may contain mesodermal components such as varying proportions of fibrous tissues and blood vessels. Fibrolipoma is an uncommon variant of lipoma and comprises a high proportion of fibrous components. An intrathoracic fibrolipoma is extremely rare; to the best of our knowledge, only three such cases have been reported till date. CASE PRESENTATION: A 51-year-old female presented with a left intrathoracic mass, which was confirmed to be a lipomatous tumor using computed tomography. A pedunculated tumor originating from the parietal pleura was resected using complete thoracoscopic surgery. Pathological examination indicated a diagnosis of fibrolipoma. Intrathoracic fibrolipomas are extremely rare; this is one of the first reported cases of successfully resecting an intrathoracic fibrolipoma using complete thoracoscopic surgery. CONCLUSIONS: The tumor was asymptomatic and relatively small when detected during a medical checkup. This enabled the successful resection of the tumor via complete thoracoscopic surgery. Although fibrolipomas are histologically benign, careful observation and follow-up are essential owing to the possibility of recurrence.

Expression of cytochrome P450 mRNAs in Type II alveolar cells from subjects with chronic obstructive pulmonary disease.

Inhaled drugs are critical for the treatment of inflammatory airway diseases such as chronic obstructive pulmonary disease (COPD). To develop better therapeutics for pulmonary disease it is of potential importance to understand molecular mechanisms of local biotransformation in the lung. Alveolar epithelial type II (ATII) cells have a key role in homeostasis in the lung, but little is known about expression patterns of genes encoding cytochrome P450 (CYP) enzymes in ATII cells. In addition, alteration of CYP gene expression has not been fully defined in COPD. We previously established a method to purify ATII cells from the adult human lung using fluorescence-activated cell sorting. By employing this technique we determined gene expression patterns of 14 CYP enzymes in ATII cells from nonsmokers (n = 4) and smokers (n = 4), both having normal pulmonary function. Although most CYP genes are highly expressed in primary hepatocytes, we found that CYP1B1 mRNA expression was 7.2-fold higher in ATII compared to hepatocytes (P = .0275). Additionally we noted a 3.0-fold upregulation of CYP2C19 and 50% reduction in CYP2J2 mRNA expressions in ATII cells isolated from patients with COPD (n = 3) compared to smokers without COPD (n = 4). These data, for the first time, detail a comprehensive set of genes encoding CYP enzymes in human ATII cells and highlights differentially expressed CYP mRNAs of patients with COPD. Such understanding may have important implications for the development of novel inhaled drugs.

A case of primary pulmonary choriocarcinoma successfully treated by surgery.

Primary pulmonary choriocarcinoma is a very rare tumor and eventually diagnosed by pulmonary resection. A poor prognosis has been reported for this disease, but appropriate treatment has not been established. Here, we report the successful surgical treatment for a primary pulmonary choriocarcinoma. A 70-year-old male presented with cough and a 3.8-cm solid tumor was detected in the right upper lobe of the lung on CT scan. Results of an open lung biopsy indicated carcinoma with indeterminate histological subtype. Right upper lobectomy was performed and the permanent pathological diagnosis was choriocarcinoma. Right after surgery, systemic and genital screening was performed by urologist, but no abnormal findings were observed. Elevation of serum ß-hCG was confirmed at the same time and the value dropped within normal range in 2 months. According to diagnosis criteria, we reached final diagnosis of primary pulmonary choriocarcinoma. The patient has been alive with no recurrence for 2 years after surgery.

Novel Monoclonal Antibody LpMab-17 Developed by CasMab Technology Distinguishes Human Podoplanin from Monkey Podoplanin.

Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain in its N-terminus. Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for the binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. Although many anti-PDPN monoclonal antibodies (mAbs) have been established, almost all mAbs bind to PLAG domains. We recently established CasMab technology to produce mAbs against membranous proteins. Using CasMab technology, we produced a novel anti-PDPN mAb, LpMab-17, which binds to non-PLAG domains. LpMab-17 clearly detected endogenous PDPN of cancer cells and normal cells in Western-blot, flow cytometry, and immunohistochemistry. LpMab-17 recognized glycan-deficient PDPN in flow cytometry, indicating that the interaction between LpMab-17 and PDPN is independent of its glycosylation. The minimum epitope of LpMab-17 was identified as Gly77-Asp82 of PDPN using enzyme-linked immunosorbent assay. Of interest, LpMab-17 did not bind to monkey PDPN, whereas the homology is 94% between human PDPN and monkey PDPN, indicating that the epitope of LpMab-17 is unique compared with the other anti-PDPN mAbs. The combination of different epitope-possessing mAbs could be advantageous for the PDPN-targeting diagnosis or therapy.

Histone deacetylase inhibitor restores surfactant protein-C expression in alveolar-epithelial type II cells and attenuates bleomycin-induced pulmonary fibrosis in vivo.

AIM: Surfactant protein-C (SP-C) of alveolar epithelial type II cells (ATII) plays a key role in maintaining alveolar integrity and repair. Mutations or decreased expression of SFTPC, the gene encoding SP-C, causes ATII injury and aberrant repair of the lung tissue to develop pulmonary fibrosis. Histone deacetylases (HDACs) epigenetically remove acetyl groups from acetylated histones and regulate transcription. HDAC inhibitors attenuated epithelial-to-mesenchymal transition (EMT) and fibrotic disorders. The aim of this study is to investigate whether Trichostatin A (TSA), a pan-HDAC inhibitor, epigenetically exerts a protective effect on ATII against fibrotic changes via the restoration of SFTPC expression. MATERIALS AND METHODS: We treated A549 cells with TGF-ß1 to induce EMT, followed by TSA treatment. We evaluated SFTPC mRNA, histone acetylation levels in the SFTPC gene promoter region, and pro-SP-C protein. C57BL6/J mice were treated with intratracheal bleomycin instillation followed by TSA administration. Histological changes and Sftpc mRNA expression in isolated ATII were evaluated. RESULTS: TGF-ß1 treatment decreased SFTPC mRNA in A549 cells. TSA restored SFTPC mRNA, and increased histone H4 acetylation in the SFTPC promoter region in vitro. The administration of TSA partially attenuated BLM-induced pulmonary fibrosis and increased the Sftpc mRNA expression in isolated ATII from bleomycin-treated lungs in vivo. CONCLUSIONS: Decreased expression of SFTPC by TGF-ß1 treatment was restored by TSA via hyperacetylation of histone H4 in the promoter region. TSA partially attenuated pulmonary fibrosis and increased Sftpc mRNA in ATII. Our findings suggest that the epigenetic restoration of SP-C would be a therapeutic target for pulmonary fibrosis.

The Role of Lysophosphatidic Acid on Airway Epithelial Cell Denudation in a Murine Heterotopic Tracheal Transplant Model.

BACKGROUND: Chronic rejection is the major leading cause of morbidity and mortality after lung transplantation. Obliterative bronchiolitis (OB), a fibroproliferative disorder of the small airways, is the main manifestation of chronic lung allograft rejection. However, there is currently no treatment for the disease. We hypothesized that lysophosphatidic acid (LPA) participates in the progression of OB. The aim of this study was to reveal the involvement of LPA on the lesion of OB. METHODS: Ki16198, an antagonist specifically for LPA1 and LPA3, was daily administered into the heterotopic tracheal transplant model mice at the day of transplantation. At days 10 and 28, the allografts were isolated and evaluated histologically. The messenger RNA levels of LPAR in microdissected mouse airway regions were assessed to reveal localization of lysophosphatidic acid receptors. The human airway epithelial cell was used to evaluate the mechanism of LPA-induced suppression of cell adhesion to the extracellular matrix (ECM). RESULTS: The administration of Ki16198 attenuated airway epithelial cell loss in the allograft at day 10. Messenger RNAs of LPA1 and LPA3 were detected in the airway epithelial cells of the mice. Lysophosphatidic acid inhibited the attachment of human airway epithelial cells to the ECM and induced cell detachment from the ECM, which was mediated by LPA1 and Rho-kinase pathway. However, Ki16198 did not prevent obliteration of allograft at day 28. CONCLUSIONS: The LPA signaling is involved in the status of epithelial cells by distinct contribution in 2 different phases of the OB lesion. This finding suggests a role of LPA in the pathogenesis of OB.

Lung squamous cell carcinoma arising in a patient with adult-onset recurrent respiratory papillomatosis.

A 75-year-old male was admitted to our hospital in December 2011 with a mass in the right upper pulmonary lobe. He was incidentally diagnosed as having tracheal papillomas 10 years ago. Bronchoscopy revealed multiple polypoid papillomas in the dorsal lesion of the trachea. Polymerase chain reaction amplification detected human papillomavirus type 11 DNA in the papilloma tissues. A computed tomography scan demonstrated the occlusion of the right superior segment bronchus with distal consolidation. Furthermore, F-18 fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography showed intense tracer uptake in the right superior segment of the lung. He underwent a right upper lobectomy. The tumor was seen as a rounded nodule, ≈ 2 cm in diameter. Histological examination of the tumor revealed squamous papilloma with papillary and solid architecture surrounded by accumulation of acute inflammatory cells. Furthermore, in a part of the tumor, squamous cell carcinoma was also present. The lymph nodes were free of tumor. After the surgery, he continued to undergo endoscopic microwave resection. Recurrent respiratory papillomatosis is a rare disease that can cause life-threatening airway compromise and malignant transformation. The present case indicates that F-18 fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography is indispensable for early detection of lung cancer arising in a patient with recurrent respiratory papillomatosis.

[Reconstruction of the posterior mitral annulus using a crescent-shaped collar; a novel technique for mitral valve replacement in the severely calcified annulus; report of a case].

A 74-year-old female who had been on hemodialysis for 4 years developed congestive heart failure. Severe mitral valve stenosis and tricuspid regurgitation were thought to be causes of her heart failure. She underwent a replacement of severely calcified mitral valve and an annuloplasty of tricuspid valve. We excised both mitral leaflets, however, just a 21 mm sizer could pass through the annulus. Because posterior annular calcification was so severe we could not palace sutures. Therefore, we placed noneverted horizontal mattress sutures from the left ventricle to the atrium on the anterior annulus, and everted horizontal mattress sutures on the posterior left atrial wall. A crescent-shaped felt patch was sutured to the posterior left atrial wall for annular reconstruction. Then the prosthetic valve was fixed onto this felt patch, resulting a 25 mm ATS valve was seated successfully at a supraannular position. Although the patient died from a sepsis on postoperative day 14, the prosthetic valve functioned normally without any perivalvular leakage.

Rapidly progressive descending aortic pseudoaneurysm resulting from primary malignant fibrous histiocytoma.

A 66-year-old man with a history of asymptomatic bilateral swelling of the axillary lymph nodes was admitted with hoarseness and pain of the right knee. Computed tomography showed a large pseudoaneurysm deriving from descending aorta. We planned to resect the pseudoaneurysm, and repair the aortic wall. Resection of the pseudoaneurysm and patch-closure of the defect of the aorta were performed successfully. Pathological examination of the pseudoaneurysm revealed malignant fibrous histiocytoma. The clinical course was very miserable. Intrathoracic recurrence occurred 7 months after surgery, and he died of multiple brain metastases 10 months postoperatively.

[Surgical outcome for small-sized non-small-cell lung cancer].

We reviewed surgical outcome for small-sized non-small-cell lung cancer. From 2004 to 2007, 109 patients with complete resected non-small-cell lung cancer of 2 cm or less in diameter were examined retrospectively. Sixty-five were male and 44 were female. Ages ranged 41~87 (median 68) years.Overall 5-year survival rate(OS) was 89%, and 5-year disease free survival rate(DFS) was 82%.Ground-glass opacity( GGO) dominant group[ GGO ratio ≥ 50% in high resolution computed tomography(CT)] and lymph node (LN) negative group were showed favorable DFS significantly. Other parameters,such as histology, tumor diameter, serum carcinoembryonic antigen (CEA) level, pleural invasion,vascular permeation, operative procedure, were showed no significant difference about DFS. All GGO dominant group patients(n=23) had no LN involvement and survived in disease free. Cases with LN involvement [n=10 (9%)] were all in solid group(GGO<50%) and their tumor diameter were over 10 mm. Sublober resections were performed in 28 cases( segmentectomy in 12, wedge resection in 16). Among these cases, intentional limited resections were done in 13 patients with GGO dominant group (tumor diameter ranged 7 to 20 mm), and they had no local recurrence. Prognosis of small-sized GGO dominant lung cancer is very favorable, so limited resection will be appropriate. But solid lesions, especially over 10 mm in tumor diameter, have possibility of LN involvement, lobectomy should be considered.

Successful weaning from a left ventricular assist device after surgical repair of a left ventricular free wall rupture.

We present a case of an insertion of a left ventricular assist device for severe cardiac failure after the repair of a left ventricular free wall rupture. A 72-year-old man was admitted with chest pain and unconsciousness, and required emergency surgical repair of a left ventricular free wall rupture under percutaneous cardiopulmonary support. Severe cardiac failure occurred postoperatively, and weaning from percutaneous cardiopulmonary support was impossible. We implanted a left ventricular assist device, and this could be removed at one week after implantation. The left ventricular assist device was very useful as a "bridge to recovery".