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Activation of Antioxidant-Response Element (ARE), Mitogen- Activated Protein Kinases (MAPKs) and Caspases by Major Green Tea Polyphenol Components during Cell Survival and Death

  • Chen, Chi;Yu, Rong;Owuor, Edward D.;Kong, A.NTony
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.605-612
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    • 2000
  • Green tea polyphenols (GTP) have been demonstrated to suppress tumorigenesis in several chemical-induced animal carcinogenesis models, and predicted as promising chemopreventive agents in human. Recent studies of GTP extracts showed the involvement of mitogen-activated protein kinases (MAPKs) in the regulation of Phase II enzymes gene expression and induction of apoptosis. In the current work we compared the biological actions of five green tea catechins: (1) induction of ARE reporter gene, (2) activation of MAP kinases, (3) cytotoxicity in human hepatoma HepG2-C8 cells, and (4) caspase activation in human cervical squamous carcinoma HeLa cells. For the induction of phase IIgene assay, (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) potently induced antioxidant response element (ARE)-mediated luciferase activity, with induction observed at 25 $\mu\textrm{m}$with EGCG. The induction of ARE reporter gene appears to be structurally related to the 3-gallate group. Comparing the activation of MAPK by the five polyphenols, only EGCG showed potent activation of all three MAPKs (ERK, JNK and p38) in a dose- and time-dependent manner, whereas EGC activated ERK and p38. In the concentration range of 25 $\mu\textrm{m}$ to 1 mM, EGCG and ECG strongly suppressed HepG2-ARE-C8 cell-growth. To elucidate the mechanisms of green tea polyphenol-induced apoptosis, we measured the activation of an important cell death protein, caspase-3 induced by EGCG, and found that caspase-3 was activated in a dose- and time-dependent manner. Interestingly, the activation of caspase-3 was a relatively late event (peaked at 16 h), whereas activation of MAPKs was much earlier (peaked at 2 h). It is possible, that at low concentrations of EGCG, activation of MAPK leads to ARE-mediated gene expression including phase II detoxifying enzymes. Whereas at higher concentrations of EGCG, sustained activation of MAPKs such as JNK leads to apoptosis. These mechanisms are currently under investigation in our laboratory. As the most abundant catechin in GTP extract, we found that EGCG potently induced ARE-mediated gene expression, activated MAP kinase pathway, stimulated caspase-3 activity, and induced apoptosis. These mechanisms together with others, may contribute to the overall chemopreventive function of EGCG itself as well as the GTP.

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Histone Deacetylase Inhibitor Stimulate CYP3A4 Proximal Promoter Activity in HepG2 Cells

  • Kim Ja Young;Ahn Mee Ryung;Kim Dae-Kee;Sheen Yhun Yhong
    • Archives of Pharmacal Research
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    • v.27 no.4
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    • pp.407-414
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    • 2004
  • The expression of CYP3A4 gene is induced by a variety of structurally unrelated xenobiotics including the antibiotic rifampicin, pregnenolone 16-carbonitrile (PCN), and endogenous hormones, that might mediate through steroid and xenobiotic receptor (SXR) system. The molecular mechanisms underlying regulation of CYP3A4 gene expression have not been understood. In order to gain the insight of the molecular mechanism of CYP3A4 gene expression, study has been undertaken to investigate if the histone deacetylation is involved in the regulation of CYP3A4 gene expression by proximal promoter in human hepatoma HepG2 cells. Also we have investigated to see if SXR is involved in the regulation of CYP3A4 proximal promoter activity in human hepatoma HepG2 cells. HepG2 cells were transfected with a plasmid PCYP3A4-Luc containing ${\~}1kb$ of the CYP3A4 proximal promoter region (-863 to +64 bp) in front of a reporter gene, luciferase, in the presence or absence of pSAP-SXR. In HepG2 cells, CYP3A4 inducers, such as rifampicin, PCN and RU486 showed minimal stimulation of CYP3A4 proximal promoter activity in the absence of SXR and histone deacetylase (HDAC) inhibitors. 4-Dimethylamino-H-[4-(2-hydroxycarbamoylvinyl)benzyl]benzamide (IN2001), a new class HDAC inhibitor significantly increased CYP3A4 proximal promoter activity over untreated control cells and rifampicin concomitant treatment with IN2001 increased further CYP3A4 proximal promoter activity that was stimulated by IN2001 The results of this study demon-strated that both HDAC inhibitors and SXR are essential to increase of CYP3A4 proximal promoter activity by CYP3A4 inducers such as PCN, rifampicin, and RU486. Especially SXR seems to be important for the dose dependent response of CYP3A4 inducing chemicals to stimulate CYP3A4 proximal promoter activity. Also this data suggested that HDAC inhibitors seemed to facilitate the CYP3A4 proximal promoter to be activated by chemicals.

Anthropometry of Nose in Korean Twenties (20대 한국인 코의 계측치)

  • Bae, Tae Hui;Yu, Young Il;Kim, Woo Seob;Kim, Han Koo
    • Archives of Craniofacial Surgery
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    • v.10 no.2
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    • pp.61-66
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    • 2009
  • Purpose: The aim of this study is to establish anatomical dimension of the nose and nostril shape in Korean twenties. Methods: Authors measured anthropometry of nose in Korean youths 1,000 individuals (male 363, female 637) and analyzed stastically them. Results: The mean measurements are as follows. 1. The shape of nostrils was classified into 7 types by the angle between both long axis of nostril (male / female). Type I ($<40^{\circ}$)1.7% / 1.72%, Type II ($41-70^{\circ}$) 26.17% / 29.35%, Type III ($71-100^{\circ}$) 39.94% / 38.77%, Type IV (unclassified) 3.85% / 5.80%, Type V ($101-130^{\circ}$) 20.93% / 17.89%, Type VI ($131-180^{\circ}$) 7.43% / 5.96%, Type VII ($>180^{\circ}$) 0% / 0.47% 2. Lengths (male/female) Width of nasal root: $25.29{\pm}2.25mm/24.72{\pm}2.40mm$ Width of nose: $37.63{\pm}2.46mm/34.77{\pm}2.11mm$ Width of columella: $7.18{\pm}0.92mm/6.92{\pm}0.86mm$ Width of alar: $4.99{\pm}1.00mm/4.74{\pm}0.91mm$ Width of nostril floor: $10.98{\pm}1.40mm/10.13{\pm}1.73mm$ Protrusion of nasal tip: $17.12{\pm}1.95mm/16.88{\pm}1.84mm$ Length of alar: $27.10{\pm}2.21mm/24.66{\pm}2.18mm$ Length of nasal root: $17.37{\pm}2.51mm/16.08{\pm}2.90mm$ Depth of nasal root: $7.83{\pm}1.63mm/6.82{\pm}1.36mm$ Length of columella: $8.13{\pm}1.40mm/7.30{\pm}1.46mm$ Height of nose: $60.50{\pm}8.90mm/59.14{\pm}9.22mm$ Height of nasal bridge: $52.68{\pm}7.49mm/50.57{\pm}7.71mm$ 3. Angles Nasofacial angle: $30.19{\pm}3.43^{\circ}/29.13{\pm}6.31^{\circ}$ Nasofrontal angle: $134.88{\pm}7.25^{\circ}/139.94{\pm}6.33^{\circ}$ Nasolabial angle: $95.08{\pm}8.95^{\circ}/95.80{\pm}8.93^{\circ}$ 4. Nasal index: $72.60{\pm}9.57%/68.21{\pm}7.03%$ Nasal tip protrusion-nasal height index: $45.64{\pm}5.21%/47.09{\pm}5.21%$ Nasal tip protrusion-nasal width index: $32.61{\pm}6.83%/31.63{\pm}6.71%$ Conclusion: These data could be useful reference for anthropometry of nose in Korean twenties.

A Retrospective Study of Ear Reconstruction of Microtia (소이증 환자의 이개재건에 관한 후향적 연구)

  • Kim, Seok-Kwun;Moon, In-Sun;Kwon, Yong-Seok;Lee, Keun-Cheol
    • Archives of Craniofacial Surgery
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    • v.10 no.2
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    • pp.109-113
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    • 2009
  • Purpose: Materials for ear reconstruction are autogeneous cartilage and artificial implants. Despite their potential for donor site complications, autogeneous cartilage frameworks remain the accepted standard for external ear reconstruction. The purposes of this study were to investigate our ear reconstruction cases for 12 years. Methods: During twelve years from January 1996 to December 2008, 70 patients visited our hospital for ear reconstruction of microtia. Among them, 65 cases used autogenous cartilage frameworks, 3 cases used tissue expander and 2 cases used artificial implants. We investigated sex & age, common site, combined malformation, operation methods & their complications, donor site & their complications, anterior chest wall deformity and aesthetic evaluation. Results: Males were affected more often than females. Male to female ratio was 1.8 : 1. The common site of microtia was right ear (72%). And hemifacial microsomia was the most common associated congenital malformation. Surgical techniques included Tanzer-Ruecker method, Tissue-expander and Artificial framework ($Medpore^{(R)}$). Incidence of complication was higher with Tissue-expander & artificial framework than with Tanzer-Ruecker method. But There are few reports of using Tissue-expander & artificial framework and there was no acceptable statistical difference. And costal cartilage was harvested in ipsilateral side and anterior chest wall deformity reported only 2 cases under 10 year-old patients. Other minor complications reported such as, wound disruption and chest wall scar, but without any serious complications. Aesthetic result was evaluated by surgeons and patients for postoperative satisfaction and clinical evaluation. Conclusion: In ear reconstruction of microtia patient, delicate surgical strategy is important for natural shape and maintenance of postoperative contour. However, many methods were introduced for reconstrucion of microtia, the authors reconstructed an auricle in Tanzer-Ruecker method, Tissue-expander and Artificial famework ($Medpore^{(R)}$) for 70 patients. In our study, we generally chose Tanzer-Ruecker method and this treatment modality was satisfactory for patients and the postoperative result was acceptable for surgeons.

Reconstruction of the Thumb, the Second and Third Finger in Patient with Amputation of Right Five Fingers (우측 수부의 모든 수지 절단 환자에서 무지와 제 2, 3 수지 재건술)

  • Lee, Jun-Mo;Kim, Gyu-Hyung
    • Archives of Reconstructive Microsurgery
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    • v.10 no.2
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    • pp.143-148
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    • 2001
  • The hand with amputation of all fingers is useless for activities of daily living and traumatic amputation of some of the fingers can result in the diminished ability to perform power grip and precision grip which is vital to maintain normal function of the hand. Precision grip is used to hold an object between the opposable thumb and flexed fingers. In power grip the object is held between the flexed fingers and the palm while the thumb applies the necessary counterpressure to maintain the grip on the object. A 35 year old male lost his right all fingers including thumb at the level of proximal phalanx from the pressure machinary accident. Thumb was reconstructed using wrap around flap and the second and third fingers were reconstructed using the second and third toe transplantation. Seven years after reconstruction, he uses the reconstructed thumb and the second and the third fingers for eating meals, writing down a paper with a pencil and putting on socks.

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Transplantation of the Neurosensory Free Flaps to the Hand (수부에 시행한 신경감각 유리 조직 이식술)

  • Lee, Jun-Mo;Lee, Ju-Hong
    • Archives of Reconstructive Microsurgery
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    • v.9 no.2
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    • pp.120-126
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    • 2000
  • Microsurgical reconstruction of the hand demands recovery of the sensation of the reconstructed free flap as well as microsurgeon's intelligence, technique and experience. Even with adequate soft tissue coverage and skeletal mobility, an insensate hand is prone to further injury and is unlikely to be useful to the patients. Authors have performed 8 cases of neurosensory free flaps in the hand, 4 cases of wrap around, 3 dorsalis pedis and 1 lateral arm flap, from July 1992 through June 1999 and followed up average 4 years and 4 months. Wrap around flap was performed for reconstruction of 4 cases of thumb, repairing deep peroneal nerve and superficial radial nerve by epineurial neurorrhaphy, and followed up for average 3 years and 10 months and calculated 9mm in the static 2 point discrimination test. Dorsalis pedis flap were 3 cases for reconstruction of the ray amputation, extensor tendon exposure and wrist exposure. Deep peroneal nerve and branch of the ulnar nerve was repaired by epineurial neurorrhaphy calculating 6mm and superficial peroneal nerve and superficial radial nerve averaging 18mm in the static 2 point discrimination test for follow up average 2 years and 9 months. Lateral arm flap was 1 case for reconstruction of the ray amputation in the hand repairing posterior cutaneous nerve to the arm to the superficial radial nerve calculating 20mm for follow up 6 years and 8 months.

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Characterization of the Expression of PKCα(Isoform) in DMH-induced Vascular Endothelial Proliferation (DMH에 의한 비정상적인 혈관 내피세포의 증식에서 Protein Kinase C 동종효소 Alpha 단백발현의 특성)

  • Nam, Su Bong;Bae, Yong Chan;Park, Suk Young;Choi, Soo Jong
    • Archives of Plastic Surgery
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    • v.34 no.6
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    • pp.679-684
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    • 2007
  • Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

The Usefulness of Transcutaneous Tissue Oxygen Pressure ($TcpO_2$) for Determination of the Point of Time at Venous Revascularization (적절한 정맥 재혈류 시점의 판단을 위한 조직산소분압의 유효성)

  • Lee, Eun-Sang;Choung, Jong-Pil;Park, Seung-Ha;Lee, Byung-Il
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.547-554
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    • 2010
  • Purpose: The purpose of this study is to delineate the optimal time of venous revascularization for preventing the flap necrosis due to venous occlusion, and to clarify the usefulness of tissue oxygen pressure ($TcpO_2$) in the determination of the point of time for venous revascularization. Methods: Thirty-six, $3{\times}3\;cm$ sized epigastric island flap was elevated in left abdomen of male Sprague-Dawley rat weighing 250 gram. Flaps were randomly assigned to six groups of six flaps according to the duration of venous occlusion with microvascular clamp; 10 minutes in the group I as the control, 60 minutes in the group II, 2 hours in the group III, 3 hours in the group IV, 4 hours in the group V, and 6 hours in the group VI, respectively. Just before removal of clamp after flap was reposed in situ, the ratio of $TcpO_2$ (tissue oxygen pressure) of the island flap to that of right abdomen was calculated in each group, and tissue specimen was harvested from the distal area of the flap for histological evaluation of vascular change. Five days later, survival area of the flap was estimated, and evaluated the correlation between the tissue oxygen pressure and the rate of flap survival. Results: The $TcpO_2$ and the survival rate of flap were decreased proportionally with the duration of venous occlusion. The ratio of the $TcpO_2$ of the flap is decreased abruptly to below sixty percentile compared to the $TcpO_2$ of normal tissue, and the survived area of the flap is decreased to nine-tenth of the designed size after three hours of total venous occlusion. Histologically, the number of congested vessels was increased according to venous occluded time, and proportionally increased after 3-hours of occlusion significantly. Conclusion: There is a close correlation between the $TcpO_2$ and the survival rate of flaps according to the duration of venous occlusion. Therefore, the $TcpO_2$ represents the hemodynamic changes within the flap, and thought to be an alternative effective tool in the flap monitoring for venous revascularization.

120 Cases of Facial Foreign Body Granuloma (안면부 이물 육아종 120례의 치험례)

  • Park, Tae-Hwan;Seo, Sang-Won;Kim, June-Kyu;Chang, Choong-Hyun
    • Archives of Plastic Surgery
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    • v.37 no.3
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    • pp.233-238
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    • 2010
  • Purpose: As the use of soft tissue fillers becomes more popular, complications such as foreign body granuloma (FBG) are increasing. We report 120 cases of facial FBG and review the available literatures. Methods: 120 patients of facial FBG in our clinic from Mar. 2003 to Feb. 2008 were complied and analyzed. A retrospective chart review was done and patient satisfaction was evaluated with a questionnaire using 5 score scale. Patients with severe inflammation sign or bizarre deformity underwent surgical excision and those with minimal symptoms or a history of hyaluronic acid injection received injection therapy using hyaluronidase. Results: 100 females and 20 males were observed. The average age was 43.7 years (from 16 to 74). 84 patients received surgical therapy and 36, injection therapy. Deformity of facial contour, foreign body sensation and inflammation sign were the three main chief complaints. 84 patients did not know what the injection materials were. The known materials are as follows: collagen, hyaluronic acid, silicone oil, paraffin. 92 cases were performed by unlicensed practitioners, 29 by physicians. Anatomical site most frequently affected by the foreign body granuloma was the cheek (25.8%), followed by forehead (19.2%), lips (15.8%), nose (9.2%), mentum (8.3%), eyelid and eyebrow (4.3%) and temple (0.8%). In 21 patients (17.5%), FBGs were found on multiple sites. Patients with inflammation sign got the highest satisfaction ($3.19{\pm}0.73$) (p=0.001) among 3 chief complaints. And patient satisfaction was statistically higher in surgical therapy group ($3.43{\pm}0.72$) than in injection therapy group ($2.97{\pm}0.88$) (p=0.003). Conclusion: We suggest that it may be beneficial to tailor the type of treatment for FBG relying on wound state and patient's chief complaints. In surgical therapy, resolute approach is necessary to correct facial deformity definitely and to minimize inflammation. Injection therapy could be another option for those with minimal symptoms or a history of hyaluronic acid injection. To prevent foreign body granuloma, not only plastic surgeons but also other physicians should inject soft tissue fillers with great caution and we should warn the public of disastrous consequences associated with illegal medical practice.

The Results of Surgical Treatments in the Peripheral Nerve Injuries (말초신경 손상 후 수술적 치료에 대한 고찰)

  • Chung, Moon-Sang;Park, Jin-Soo;Seo, Joong-Bae;Park, Yong-Bum
    • Archives of Reconstructive Microsurgery
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    • v.5 no.1
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    • pp.121-127
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    • 1996
  • Peripheral nerve injury occurs mostly by trauma and is usually associated with fracture of bone and joint, muscular injury and tendon injury and it also evokes paralysis and anesthesia. When treatment of peripheral nerve injury is considered,, the modality of treatment is decided by the general condition of the patient, type of injury, associated injuries and the condition of wound. To get the maximum results, surgical treatment and reconstruction and rehabilitation should all go in hand-in-hand. From January 1985 to December 1994, we observed 61 patients that had operation without reconstruction due to peripheral nerve injury with a follow-up period of more than 1 year. Among the 61 patients, 44 were men(72%) and 17 were women(28%). Follow-up period was average 19 months. Age distribution was mostly in their twenties with a mean age of 28 years. Time interval of operation after injury was average 11 months. Trauma was the main cause of peripheral nerve injuries with a proportion of 87%. 31 patients had neurorrhaphy, in which case 14 patients had stay suture and 17 patients did not. 14 patients had nerve graft, and 16 patients had neurolysis. We used our scales to compare the results of surgery on the basis of British Research Council System. We gave scores to every sensory and motor scale to estimate functional improvement and emphasized on motor functional improvement. The total score = sensory score + ($2{\times}motor$ score). We considered 8-9 points as excellent, 6-7 points as good, 2-5 points as fair, 0-1 points as poor result. We considered excellent and good as much improved. Excellent and good results were obtained in 13 out of 14 neurorrhaphy with stay suture(93%), 12 out of 17 neurorrhaphy without stay suture(71%), 6 out of 14 nerve graft(43%), 12 out of 16 neurolysis(75%). Among the patients with neurorrhaphy done within 3 months, 11 out of 14(86%) showed improvement, but among the patients after 4 months 3 out of 17(76%) showed improvement. 84% of improvement was observed in the patients with time interval from injury to surgery within 3 months, and 64% in the patients with time interval after 4 months. In the aspect of age, 77% with the age below 20 years, 70% with the age between 21 and 30 years, 66% with the age above 31 years showed improvement. We conclude that considering degree of injury, time interval from injury and age with the adequate method of treatment, we can obtain good results from surgery.

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