• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권3호
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• pp.199-206
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• 2010

This study investigated the effects of strontium on osteoblastic phenotypes of cultured human periostealderived cells. Periosteal tissues were harvested from mandible during surgical extraction of lower impacted third molar. Periosteal-derived cells were introduced into cell culture. After passage 3, the periostealderived cells were further cultured for 28 days in an osteogenic induction DMEM medium supplemented with fetal bovine serum, ascorbic acid 2-phosphate, dexamethasone and at a density of $3{\times}10^4$ cells/well in a 6-well plate. In this culture medium, strontium at different concentrations (1, 5, 10, and 100 ${\mu}g$/mL) was added. The medium was changed every 3 days during the incubation period. We examined the cellular proliferation, histochemical detection and biochemical measurements of alkaline phosphatase (ALP), the RT-PCR analysis for ALP and osteocalcin, and von Kossa staining and calcium contents in the periostealderived cells. Cell proliferation was not associated with the addition of strontium in periosteal-derived cells. The ALP activity in the periosteal-derived cells was higher in 5, 10, and 100 ${\mu}g$/ml strontium-treated cells than in untreated cells at day 14 of culture. Among the strontium-treated cells, the ALP activity was appreciably higher in 100 ${\mu}g$/ml strontium-treated cells than in 5 and 10 ${\mu}g$/ml strontium-treated cells. The levels of ALP and osteocalcin mRNA in the periosteal-derived cells was also higher in strontium-treated cells than in untreated cells at day 14 of culture. Their levels were increased in a dose-dependent manner. Von Kossa-positive mineralization nodules were strongly observed in the 1 ${\mu}g$/ml strontium-treated cells at day 21 and 28 of culture. The calcium content in the periosteal-derived cells was also higher in 1 ${\mu}g$/ml strontium-treated cells at day 28 of culture. These results suggest that low concentration of strontium stimulates the osteoblastic phenotypes of more differentiated periosteal-derived cells, whereas high concentration of strontium stimulates the osteoblastic phenotypes of less differentiated periosteal-derived cells. The effects of strontium on osteoblastic phenotypes of periosteal-derived cells appear to be associated with differentiation-extent.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제28권5호
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• pp.375-395
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• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• 대한간호학회지
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• 제5권2호
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• pp.23-28
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• 1975

One hundred and fifty seven cesarean sectioned patients among 2010 deliveries, were reviewed with the following clinical charts from January, 1973 to December, 1974 at Taegu Presby-terian medical Center. The results are as follows; 1) The section rate was 7.8% 2) In age distribution, the most frequent age was 26~30ys and it~s rate was 37%, and others were as follows : 31~35ys 24.2%, 21~25ys 21%, 36~40ys 10.8%, 41~45ys 7% respective]y- 3) In indication, the cephalopelvic disproportion was the most prevalent and it's rate was 29.3%, and others were as follows : malpresentation 14% previous cesarean section 10.8% uttering dysfunction 9.6%, totemic 4%, elderly primigravida 4%, fetal distress 2.5% post maturity 3.2%, wanted cesarean section 1, 9%, pelvic deformity 1.2% twin 1.9%, abruptio placenta 1.2%, Vaginal stricture 1.2%, and others 2.5%. 4) In type of operation, tower cervical transverse type was 82.1%, classical type 17%, ana cesarean hysterectomy was 0.9% respectively. 5) General Anesthesia was major type and it's rate was 63%, and spinal anesthesia 17.2%, epidermal anesthesia 14% and other combined type was 5.8%. 6) Percental mortality was 11.4%. 7) There was no maternal death. 8) Among the cesarean section complications, wound infection was 8,4%, headache 4.4%, urinary tract infection 1.2% and one case in endometritis. 9) In combined surgery, tubal legation was 31.2%, appenectomy 7.6%, posterior repair 1.2%, salpingoop-horectomy 1.9%, and one case in resection of uttering septum. 10) Cesarean section was performed 61.3% in multipara and 38.2% in primipara. 11) 47 (30%) had received artificial abortions. 12) 28 (17.8%) had previously experienced cesarean section.

• Applied Microscopy
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• 제26권3호
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• pp.349-367
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• 1996

The development of small granule-containing cell in the superior cervical ganglion was studied by electron microscopic method in human fetuses ranging from 40 mm to 260 mm crown rump length (10 to 30 weeks of gestational age). At 40 mm fetus, the superior cervical ganglion was composed of clusters of undifferentiated cells, primitive neuroblasts, and unmyelinated nerve fibers together with blood vessels. At 90 mm fetus, the superior cervical ganglion consisted of neuroblasts, satellite cell, small granule-containing cells, and unmyelinated nerve fibers. Two morphological types of the small granule-containing cells in the superior cervical ganglion were first indentified at 90 mm fetus, but were rare. Type I granule-containing cell occurred in solitary and had long processes, whereas type II cells tend to appeared in clusters near the blood capillaries. The granule-containing cells were characterized by the presence of dense-cored vesicles ranging from $150{\sim}300nm$ in diameter in both the cell bodies and processes. Other organelles included abundant mitochondria, rough endoplasmic reticulum, neurotubules, and widely distributed ribosomes. The granule-containing cells had long processes similar to those found in principal ganglionic cells. They could be identified by their content in dense-cored vesicles. The small granule-containing cells increased somewhat in size and number with increase of fetal age. Synaptic contacts were first found on the solitary granule-containing cell at 150 mm fetus. Synaptic contacts between the soma and processes of type I granule-containing cells and preganglionic axon terminals were observed. In addition, synaptic junctions between the processes of granule-containing cells and presumed dendrite of postganglionic neuron were also observed from 150 mm onward. On the basis of these features type I granule-containing cells could be considered as interneurons. The clusters of type II granule-containing cells were located in the interstitial or subcapsular portions of the ganglion, and had short processes which ended in close relation to fenestrated capillaries. Therefore it may be infer that clusters of type II granule-containing cells have an endocrine function.

• The Korean Journal of Physiology and Pharmacology
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• 제15권6호
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• pp.363-369
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• 2011

The present study elucidated the effect of the selective inducible nitric oxide synthase (iNOS) inhibitor $N^6$-(1-iminoethyl)-L-lysine (L-NIL) on monosodium urate (MSU) crystal-induced inflammation and edema in mice feet. L-NIL (5 or 10 mg/kg/day) was administered intraperitoneally 4 h before injection of MSU (4 mg) into the soles of mice hindlimb feet. Twenty-four hours after MSU injection, foot thickness was increased by 160% and L-NIL pretreatment reduced food pad swelling in a dose dependent manner. Pretreatment of 10 mg/kg/day L-NIL significantly suppressed the foot pad swelling by MSU. Plasma level of nitric oxide (NO) metabolites and gene expression and protein level of iNOS in feet were increased by MSU, which was suppressed by L-NIL pretreatment. Similar pattern of change was observed in nitrotyrosine level. MSU increased the gene expression of tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-$1{\beta}$ and L-NIL pretreatment suppressed MSU-induced cytokines expression. The mRNA levels of superoxide dismutase and glutathione peroxidase1 were increased by MSU and L-NIL pretreatment normalized the gene expression. Phosphorylation of extracellular signal-regulated kinase 1/2 and p38 was increased by MSU, which was suppressed by L-NIL pretreatment. The mRNA levels of iNOS, TNF-${\alpha}$, and IL-$1{\beta}$ were increased by MSU in human dermal fibroblasts, C2C12 myoblasts, and human fetal osteoblasts in vitro, which was attenuated by L-NIL in a dose dependent manner. This study shows that L-NIL inhibits MSU-induced inflammation and edema in mice feet suggesting that iNOS might be involved in MSU-induced inflammation.

• IMMUNE NETWORK
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• 제5권3호
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• pp.172-178
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• 2005

Background: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. However, it is known that the induction of immune response to CEA is very difficult because CEA is a self-antigen expressed in fetal cells and weakly expressed in normal colorectal epithelial cells. To enhance anti-tumor immunity specific for CEA, recombinant CEA protein was modified using listeriolysin O (LLO) for endosomal lysis and trans activator of transcription (Tat) domain for transducing extracellular proteins into cytoplasm. Methods: After immunization using dendritic cells pulsed with Tat-CEA, both Tat-CEA and LLO, and both Tat-CEA and Tat-LLO, antibody titer to CEA and LLO, cytotoxic T lymphocyte activity and the frequency of IFN-${\gamma}$ producing T lymphocytes were measured. Results: Immunization using DC pulsed with both Tat-CEA and Tat-LLO protein showed the increasement of production of CEA-specific antibody in serum, cytotoxic T lymphocyte activity, the frequency of IFN-${\gamma}$ secreting T cells, compared with DC pulsed with both Tat-CEA and LLO. Furthermore the ratio of CD8+T cell to $CD4^+$ cell among CEA-specific T cells was increased in group pulsed with both Tat-CEA and Tat-LLO. Conclusion: These results suggested that DC vaccine using Tat-LLO could be used for the development of effective immunotherapy for the treatment of tumor.

• Molecules and Cells
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• 제40권6호
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• pp.434-439
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• 2017

Gestational diabetes mellitus (GDM), one of the common metabolic disorders of pregnancy, leads to functional alterations in various cells including stem cells as well as some abnormalities in fetal development. Perivascular stem cells (PVCs) have gained more attention in recent years, for the treatment of various diseases. However, the effect of GDM on PVC function has not been investigated. In our study, we isolated PVCs from umbilical cord of normal pregnant women and GDM patients and compared their phenotypes and function. There is no significant difference in phenotypic expression, response to bFGF exposure and adipogenic differentiation capacity between normal (N)-PVCs and GDM-PVCs. However, when compared with N-PVCs, early passage GDM-PVCs displayed decreased initial rates of cell yield and proliferation as well as a reduced ability to promote wound closure. These results suggest that maternal metabolic dysregulation during gestation can alter the function of endogenous multipotent stem cells, which may impact their therapeutic effectiveness.

• Asian Pacific Journal of Cancer Prevention
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• 제16권4호
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• pp.1609-1613
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• 2015

Purpose: To determine the rate of lymph node metastasis, oncologic and pregnancy outcomes in apparent early-stage malignant ovarian germ cell tumor (MOGCT). Materials and Methods: Medical records of apparent early-stage MOGCT patients undergoing primary surgical treatment at Siriraj Hospital, Bangkok, Thailand, between January 2006 and December 2013, were retrospectively reviewed. Results: Thirty-eight patients had apparent stage I-II MOGCT. The mean age was $22.1{\pm}7.7years$ (median, 20.8 years; range, 7.7-35.6 years). The mean tumor size was $17.8{\pm}6.5cm$ with a median of 20 (range 4-30) cm. Three most common histopathologies were dysgerminoma (12 patients, 31.6%), immature teratoma (12 patients, 31.6%), and endodermal sinus tumor (6 patients, 15.8%). Twenty-seven of 38 patients underwent lymphadenectomy; 13 patients (48.2%) were stage IA and 8 patients (29.6%) were stage IC. The rate of retroperitoneal nodes metastasis was 7.4% (2/27 patients). At 26.1 months of median follow-up time (range 1.9-88.5 months), 9 patients retained fertility functions, with uneventful pregnancies in 3 of these. Only one patient (2.6%) had progression of disease at 4.9 months after surgery. The 5-year survival rate was 97.4%. Conclusion: As the rate of pelvic or para-aortic node metastasis in MOGCT is considerable, lymphadenectomy should be incorporated in surgical staging procedures.

• Clinical and Experimental Reproductive Medicine
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• 제38권2호
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• pp.109-114
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• 2011

Peritoneal pregnancy is an implantation in the peritoneal cavity exclusive of tubal, ovarian, or intra-ligamentary implantations. This is a rare obstetric complication with high maternal mortality and even higher perinatal mortality, and secondary type was most common. Risk factors for peritoneal pregnancy are previous history of extrauterine pregnancy or tubal surgery pelvic post-inflammatory status or presence of an intrauterine device. As it is a life-threatening condition, expectant management carries a risk of sudden life-threatening intra-abdominal bleeding and a generally poor fetal prognosis. So, when it is recognized, immediate termination of pregnancy is usually recommended. Early diagnosis of peritoneal pregnancy is difficult, but is important by their life threatening progress course to patients. Recently, we experienced primary peritoneal pregnancy which meets both the original and modified criteria. In this paper, we reported the case of early diagnosed and successfully treated peritoneal pregnancy despite of their diagnosis was incidentally.

• Neonatal Medicine
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• 제15권1호
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• pp.100-104
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• 2008

난소 낭종은 신생아기 여아의 가장 흔한 복강 내 낭성 종괴로, 대부분은 자연 소멸되지만 일부는 난소 염전, 낭종 내 출혈, 난소 파열 등의 합병증으로 인해 수술적 치료를 필요로 한다. 난소 낭종의 파열은 매우 드물게 발생하는 것으로 알려져 있으며, 심한 혈성 복수나 복막염 등으로 인해 사망을 초래하기도 한다. 저자들은 산전 초음파를 통해 복강내 종괴가 발견되었고 출생 후 경도의 복부 팽만 및 두 차례의 혈뇨를 주소로 내원한 2일된 신생아에서 탐색 개복술을 통해 우측 난소 낭종의 염전 및 파열을 진단하고, 우측 난관-난소절제술을 통해 성공적으로 치료하였기에 문헌고찰과 함께 보고하는 바이다.