Study for pathogenesis of congenital cholesteatoma with comparison of proteins expressed in congenital cholesteatoma, acquired cholesteatoma and skin of the external auditory canal through proteomics

Dept. of Medicine/박사Congenital cholesteatomas are epithelial lesions which represent invasive growth and osteolysis. The incidence consists of 1-5 % in middle ear cholesteatoma, but the detection rate is increasing. Especially congenital cholesteatoma can make hearing function worse, when it is not properly managed. While the biochemical study of acquired cholesteatomas is progressed some degree, the pathogenesis of congenital cholesteatomas is nearly unknown. Therefore the biochemical study regarding it is necessary now. Considering limitations of genetic study, crossover analysis of proteins expressed in congenital cholesteatomas via proteomics and ELISA can give us some clue for the understanding of its pathogenesis and make progress in treatment for it.Four congenital cholesteatoma matrices, and four samples of normal external auditory canal skin and two acquired cholesteatoma were obtained intraoperatively. We performed 2D electrophoresis in order to separate the proteins by molecular weight and detected. We then analyzed the upregulated spots from the congenital cholesteatoma matrices comparing the acquired cholesteatoma specimens through MALDI-TOF MS and immunohistochemical staining.In the 2D electrophoresis, four samples of congenital cholesteatoma showed very similar pattern of proteins expression. Comparing protein expression of acquired cholesteatoma and congenital cholesteatoma, some proteins were simultaneously expressed and the others not. Similar and different components of protein expression between the skin and congenital cholesteatoma sample were identified. The total number of spots in the 2-DE image from congenital cholesteatoma, acquired cholesteatoma and the skin was 556, 460 and 624. In the congenital and acquired cholesteatoma 326 proteins were simultaneously expressed. Of them, 56 proteins were not expressed in the skin. In the skin and congenital cholesteatoma 373 proteins were simultaneously expressed. Therefore 127 proteins were expressed only on congenital cholesteatoma. Major spots expressed in congenital cholesteatoma gels more than in acquired cholesteatoma gels were selected and analyzed, which included titin, PRO2619, forkhead transcription activator homolog, ryanodine receptor 2, plectin 1, keratin 10 and leucine zipper protein 5. Spots which were densitometrically expressed in congenital cholesteatoma gels more than spots of acquired cholesteatoma and were not expressed in the skin gels, were analyzed. They were nine spots, which were Ig heavy chain variable region (gi951281), chain A structure of human muscle pyruvate kinase (Pkm2, gi223462173). In immunostaining assay, forkhead transcription activator homolog (FKH 5-3), keratin-10 and titin in congenital cholesteatoma specimen was identified in contrast to the negative control.In this study, we found the proteins expressed in congenital cholesteatoma more than in acquired cholesteatoma. Among them, proteins which were not expressed in the skin were found. Some of them maybe play an important role in the pathogenesis of congenital cholesteatoma. But the function study for them should be continued.34810359) and cyclic nucleotide gated channel beta 3 (gi81170715), chain B crystal structure of human caspase-2 in complex with Acetyl-Leu-Asp-Glu-Ser-Asp-Cho (gi115298657), caspase-5/f (gi327532753), protein S100-A7 (gi119612656), DNA repair and recombination protein RAD54B isoform 3 (gi67464392), ribosomal protein L35a isoform CRA_c (giope

Trans-tympanic Cartilage Chip Insertion for Intractable Patulous Eustachian Tube

Background and Objectives: Patulous Eustachian tube (PET) causes troublesome autophony. We treated PET using tragal cartilage chip insertion to fill in the concavity within the tubal valve and evaluated the feasibility of this method. Subjects and METHODS: This study used a prospective design. Eleven patients with PET disorder were included. Tragal cartilage chip insertion via a transcanal approach into the Eustachian tube (ET) was performed in 14 ears of those patients. They were followed-up for at least 12 months after surgery and were evaluated by symptom questionnaire scores. RESULTS: The average follow-up was 16.4 months. Thirteen of fourteen ears received immediate complete relief of autophony symptoms. Autophony symptoms at the last follow-up were as follows: four ears (28.6%) had complete relief; five ears (35.7%) showed satisfactory improvement; four ears (28.6%) showed significant but unsatisfactory improvement; and one ear (7.1%) was unchanged. The PET symptom questionnaire in the affected ears showed a significant reduction in autophony (p=0.047) and improvement in breathing sound conduction (p=0.047). There were no complications such as otitis media or occlusion symptom. Conclusions: Transtympanic cartilage chip insertion into the ET provides a safe and accessible surgical option for the treatment of PET.ope

Tracheal reconstruction with a free vascularized myofascial flap: preclinical investigation in a porcine model to human clinical application

Although there are various methods for tracheal reconstruction, such as a simple approximation with suturing and coverage with adjacent soft tissue or muscle, large defects >50% of the tracheal length still present a clinical challenge. Tissue engineering, a recent promising way to possibly resolve this problem, requires a long preparatory period for stem cell seeding on a scaffold and relatively invasive procedures for stem cell harvesting. As an alternative, we used a vascularized myofascial flap for tracheal reconstruction. In four porcine models, the deep inferior epigastric perforator (DIEP) was used in two and the superior epigastric artery perforator (SEAP) in two. Transformation of the surface of the transplanted myofascial flap was analyzed in the airway environment. The flaps failed in the DIEP group due to venous congestion. At 12 weeks postoperatively, none of SEAP group showed any signs of respiratory distress; the inner surface of the implant exhibited stratified squamous epithelium with sparse cilia. In the clinical setting, a patient who underwent a tracheal reconstruction with a vascularized myofascial flap and 2-year follow-up was in good health with no respiratory distress symptoms.ope

A Case of Merkel Cell Carcinoma of the External Auditory Canal Skin with Annulus and Tympanic Membrane

Merkel cell carcinoma is an extremely rare neuroendocrine neoplasm of the external auditory canal (EAC) skin, which requires wide excision and adjuvant radiation due to a high recurrence rate. In this report, we describe a case of Merkel cell carcinoma arising from the EAC which was successfully treated with endoscopic excision. A 32-years-old female patient with a history of papillary thyroid cancer was diagnosed with an EAC tumor incidentally. There was a erythematous papular lesion on the posterior EAC without any evidence of locoregional metastasis. The patient underwent endoscopic tumor resection under general anesthesia, and the final pathologic report confirmed the diagnosis of Merkel cell carcinoma. There was no clinical or radiographic evidence of recurrence or metastasis of Merkel cell tumor for 41 months after surgical resection alone.ope

The Time Course of Monocytes Infiltration After Acoustic Overstimulation

Cochlea macrophages regulate cochlea inflammation and may harbors the potentials to protect hearing function from injury, including acoustic overstimulation. Cochlea macrophage numbers increase at 3-7 days after acoustic stimulation. However, the exact timing of macrophage infiltration and maturation from inflammatory monocytes is unclear. Furthermore, neutrophils may also be involved in this process. Therefore, in this study, we investigated time-dependent immune cell infiltration, macrophage transformation, and neutrophil involvement following acoustic stimulation. Flow cytometry and immunofluorescence were conducted in C-X3-C motif chemokine receptor 1 (CX3CR1)+/GFP mice after acoustic overstimulation (at baseline and at 1, 2, 3, and 5 days after exposure to 120 dB for 1 h) to identify inflammatory monocytes in the cochlea. RNA-sequencing and quantitative polymerase chain reaction were performed to identify differentially expressed genes. Inflammatory monocytes infiltrated into the lower portion of the lateral wall within 2 days after acoustic overstimulation (dpn), followed by transformation into macrophages at 3-5 dpn via CX3CR1 upregulation and Ly6C downregulation. In addition, inflammatory monocytes were aggregated inside the collecting venule only at 1 dpn. Neutrophils were not a major type of phagocyte during this response. The gene encoding C-C motif chemokine ligand 2 gene was significantly upregulated as early as 3 h after acoustic overstimulation. Given these results, treatment to control immune response after a noise-induced hearing loss should be applied as soon as possible.ope

Inflammatory Monocytes Infiltrate the Spiral Ligament and Migrate to the Basilar Membrane After Noise Exposure

Objectives: Noise exposure leads to an increase in the macrophage population. This increment is thought to be caused by the transformation of infiltrated monocytes into macrophages rather than by proliferation of the cochlear resident macrophages. However, studies on infiltrated monocytes in the cochlea are scarce. Thus, we aimed to investigate the infiltration of monocytes and their transformation into macrophages after noise exposure. Methods: In wild-type and CX3CR1+/GFP C57/B6 mice, inflammatory monocytes were identified by immunofluorescence of mouse cochlear cells. The findings were confirmed and quantitated by flow cytometry. Results: One day after noise exposure, monocytes were identified in the spiral ligament. Flow cytometric analysis confirmed that the monocyte population peaked on post-noise exposure day 1 and decreased thereafter. On day 3 after noise exposure, amoeboid-type macrophages increased in the crista basilaris, and on day 5, they spread to the basilar membrane. Conclusion: Infiltrated monocytes were successfully observed 1 day after noise exposure, preceding the increase in the macrophage population. This finding supports the proposal that infiltrated monocytes transform into macrophages.ope

Facial nerve management in jugular paraganglioma surgery: a literature review

Objective: This literature review analysed facial nerve management strategies in jugular paraganglioma surgery and discusses the tumour resection rate and the facial nerve outcome associated with each technique. Methods: A retrospective review of PubMed and Medline articles on the surgical treatments for jugular paraganglioma was performed. Tumour resection rates and post-operative facial nerve function after non-rerouting, short anterior rerouting and long anterior rerouting approaches were evaluated for each article. Results: A total of 15 studies involving a total of 688 patients were included. Post-operative facial nerve function was similar after non-rerouting and short anterior rerouting approaches (p = 0.169); however, both of these techniques had significantly better post-operative facial nerve outcomes compared with long anterior rerouting (p < 0.001 and p = 0.001, respectively). The total tumour removal rate was significantly higher for long anterior rerouting than with the non-rerouting approach (p = 0.016). There was no difference in total tumour removal rate between the long and short anterior rerouting approaches (p = 0.067) and between the short anterior rerouting and non-rerouting approaches (p = 0.867). Conclusion: No strict guidelines for facial nerve management in jugular paraganglioma resection are available. Although long anterior rerouting provides the best tumour exposure along with a low morbidity rate, case-by-case selection of the surgical approach is recommended

개인주의-집합주의의 개인차가 거절행동에 미치는 영향

학위논문(석사)--서울대학교 대학원 :심리학과 사회심리전공,2002.Maste

Efficacy of drum graft with tragal cartilage and the coinsertion of PORP in the intact canal wall mastoidectom

의학과/석사[한글] 폐쇄형 유양동삭개술을 시행한 후, 고막에 고막함몰이 발생하는 경우가 5-20 %까지 보고 되고 있다. 이의 방지를 위해, 많은 저자들이 연골을 사용하고 있다. 다양한 디자인의 이주연골을 이용한 고실성형술이 있지만 좀더 나은 이개연골의 디자인을 소개하고자 한다. 이주연골을 십자로 홈을 파서 가동성을 주고, 환기관 삽입 시 용이하게 하며 연골을 포함하여 고막함몰을 방지하고 또한 PORP를 삽입하여 청력의 증진을 가져오고, 고막함몰을 역시 방지하는 술식이다. 수술 전후의 고막 상태와 청력 등을 보고 기존의 유양동삭개술 후의 상태와 비교하여 본 술식의 유용성을 평가해 보고자 하였다.고막 이식으로 연골-연골막 복합체를 사용하고, 동시에 PORP를 삽입한 총 47 례를 대상으로 하였다. 이중 43 례에 대해 청력 검사가 시행되었다. 수술의 결과는 수술 전-후의 순음청력, 골기도차, 그리고 수술 후 고막 소견으로 평가하였다. 20 례가 남자, 27례가 여자였으며, 나이는 7～71세까지이다. 수술 후 평균 추적관찰 기간은 35개월이고, 수술 후 순음청력은 술후 평균 16개월 후에 시행되었다. 총 45례 중 3례 (6.7%)에서 고막함몰이 추적 관찰기간 동안 관찰되었으며 이외에 PORP가 약간 돌출이 1례가 있었고, 고막의 외측 치유(lateral healing)된 1례와 술 후 감염된 2례가 있었다. 평균 골기도차는 통계학적으로 유의하게 30.0 dB에서 24.0 dB로 (p<0.04) 감소하였고, 평균 기도청력은 통계학적으로 유의하게 47.3 dB(SD13.))에서 35.7 dB(SD15.5)로 개선되었다.(p<0.0003)이상의 결과로 생리적으로 디자인된 연골-연골막 복합체를 이용한 고실성형술과 PORP의 동시 삽입은 고막함몰 발생과 무기화를 효과적으로 줄일 수 있고, 청력의 개선도 기대할 수 있는 술식으로 사료된다. 연골의 디자인이 고막이식을 용이하게 해 주고, 고막함몰이 발생하더라도 환기관을 삽입하기 쉽고, PORP의 돌출을 방지할 수 있게 해 줄 것이다. [영문]After performing intact canal wall mastoidectomy, formation of retraction pocket has been reported in 5~20% of the cases. To prevent this, many otologists have been using cartilage. Though there were various types of tympanoplasty, we would introduce the better design of tympanoplasty with tragal cartilage. The biologically designed cartilage which had cross-shaped groove was made to facilitate the manipulation so that the graft on tympanic ring could be easily performed. The groove of cartilage can be used later as a passage for ventilation tube insertion. And the insertion of PORP result in the improvement of hearing and the prevention of retraction pocket. We would evaluate the efficacy of the new tympanoplasty by comparing the reports of usual mastoidectomy with pre- and post-operative hearing and drum findingTotal 47 cases which were performed intact canal wall mastoidectomy with the insertion of PORP and drum graft using cartilage-perichondrial flap have been reviewed. Out of these, 43 cases have been selected to audiologic test. The result of operation was evaluated by comparing pre- and post-operative pure tone audiogram, air bone gap and postoperative drum findings. Twenty subjects were male and 27 female, aged from 7 to 71 years. Average follow up period after operation was 35months, and post-operative pure tone audiometry was conducted average 16 months after operation.Out of 45 cases, retraction pocket was observed in 3 cases (6.7%). Slight protrusion of PORP and lateral healing of drum was observed in 1 case each, and there were 2 cases of postoperative infection. Also number of cases with air bone gap less than 20 dB increased from 9(23%) cases pre-operation, to 20 cases (51%) post operation. Air bone gap average statistically decreased from 30.0 dB to 24.0 dB(p<0.04), and the mean air conduction thresholds statistically improved from a preoperative level of 47.3 dB (SD13.0) to a level of 35.7 dB(SD15.5)(p<0.0003)Biologically designed cartilage tympanoplasty with the co-insertion of PORP can decrease the formation of retraction pocket and atelectasis of drum as well as improving hearing. We assumed that the new cartilage-perichondrial flap facilitates the drum graft, and even if retraction pocket is formed, it is easier to insert the ventilation tube. Also it could prevent the extrusion of PORP.ope