• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5653-5657
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• 2012

Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.

• 한국육종학회지
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• 제42권5호
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• pp.495-501
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• 2010

복숭아의 기원은 중국으로 알려져 있으며, 저장기간이 짧아 소비자들의 선호도가 낮았다. 이런 이유로 육종가들은 저장성을 높이는 것과 향과 맛을 좋게 하는데 육종 목표를 설정했으며, 국립원예특작과학원 육종가들은 이 목표에 따라 새로운 품종('천홍', '수홍', '하홍', '유명', '백미조생', '천향', '진미', '수미', '미스홍', '유미')을 육성하였다. 국립원예특작과학원 과수과에서는 육성 품종의 보호와 특허권을 확보하기 위해 분자생물학적 마커의 개발이 필요하여, 235 세트의 Operon 프라이머를 이용하여 품종 특이적인 DNA 절편 134개를 확보했고, 염기서열 분석을 통해 SCAR 마커를 개발하였다. 개발된 마커 14 세트를 이용하여 육성 품종과 육성 품종의 모본 부본이 포함된 30품종에 대해 구분이 가능하였다. 이 결과를 토대로 국립원예특작과학원 과수과에서 육성한 품종에 대한 분자생물학적 근거로 특허권을 확보하고, 묘목시장에서 품종에 대한 정확한 근거를 제시할 것으로 기대된다.

• Journal of Gastric Cancer
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• 제17권1호
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• pp.11-20
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• 2017

Purpose: Acupuncture has recently been accepted as a treatment option for managing postoperative ileus (POI) and various functional gastrointestinal disorders. Therefore, we conducted a prospective randomized study to evaluate the effect of acupuncture on POI and other surgical outcomes in patients who underwent gastric surgery. Materials and Methods: Thirty-six patients who underwent distal gastrectomy for gastric cancer from March to December 2015 were randomly assigned to acupuncture or non-acupuncture (NA) groups at 1:1 ratio. The acupuncture treatment was administered treatment once daily for 5 consecutive days starting at postoperative day 1. The primary outcome measure was the number of remnant sitz markers in the small intestine on abdominal radiograph. The secondary outcome measure was the surgical outcome, including the times to first flatus, first defecation, start of water intake, and start of soft diet, as well as length of hospital stay and laboratory findings. Results: The acupuncture group had significantly fewer remnant sitz markers in the small intestine on postoperative days 3 and 5 compared to those in the NA group. A significant difference was observed in the numbers of remnant sitz markers in the small intestine with respect to group differences by time (P<0.0001). The acupuncture group showed relatively better surgical outcomes than those in the NA group, but the differences were not statistically significant. Conclusions: In this clinical trial, acupuncture promoted the passage of sitz markers, which may reflect the possibility of reducing POI after distal gastrectomy.

• Applied Biological Chemistry
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• 제42권4호
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• pp.317-323
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• 1999

칼슘을 첨가한 메틸셀룰로오스(CaMC)와 하이드록시프로필 메틸셀룰로오스(CaHPMC-15)의 식용코팅을 미숙자두에 처리하여 자두의 연화 현상을 알아보았다. 비코팅 자두는 저장 기간 동안 중량 및 호흡률 감소 현상을 나타내었고, 적정산도는 감소하였으며, pH는 증가하였다. 그러나 CaMC와 CaHPMC-15를 코팅하였을 때 초기의 중량감소와 호흡률감소 효과를 나타내었다. 일반적으로 연화의 지표로 사용되는 수용성 펙틴 함량과 polygalacturonase(PG)의 활성은 저장 중 증가하였으며, pectin esterase(PE)의 존재 또한 확인되나 CaMC와 CaHPMC-15식용코팅에 의하여 연화현상을 억제하였다. 저장 기간 경과에 따라 기계적 경도가 감소되었으나 식용코팅에 의하여 자두의 경도 유지 효과가 있었으며, 관능검사 결과 저장 기간 중 자두 조직의 연화가 감지되었고 코팅자두의 경우 비코팅자두보다 더 단단한 것으로 평가되었다. 결과적으로 미숙자두에 CaMC와 CaHPMC-15를 코팅함으로서 수확 후 연화현상을 억제할 수 있었다.

• 미생물학회지
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• 제31권3호
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• pp.218-223
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• 1993

Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl$_{2}$ were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10$^{-5}$ . The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.384-390
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• 2006

Pleomorphic adenoma is the most common salivary benign tumor, constituting over 60% of parotid gland tumor, 25% of sublingual gland tumor and 50% of minor salivary gland tumor. It is somewhat more frequent in the fourth to sixth decades. The recurrent rate which enforces only a simple enucleation is very high $(20{\sim}45%)$. Histologically, it contains the epithelial cell, the myo-epithelial cell and mesenchymal ingredient, which is various aspect. We analyzed clinicopathologically and immunohistochemically the patients(34 cases) who are diagnosed with pleomorphic adenoma in Dept. of Oral & Maxillofacial Surgery, College of Dentisty, Dankook university since 1998. The results are as follow: 1. The incidence of the tumor was most frequent in age 30 to 50. The ratio of male to female was 1:1.43. 2. The most chief complain was a painless mass(94.1%) and the duration time was more than decade in 18 cases(52.9%). 3. Palate(soft & hard palate) was the most occurred site(64.7%). In major salivary glands, the parotid gland was the most frequent site(17.6%). 4. The tumor size was 2 to 3cm on the average. Most of tumors were with capsule(91.2%). 5. Surgical excision was a main treatment method(20 cases, 58.8%) and 14 cases were excised with a glandectomy, 1 case was treated with a partial maxillectomy. Only 1 case of all cases was recurred. 6. Histopathologically, 9 cases(26.5%) were cellular type, 11 cases(32.4%) were intermediate(classic) type and 14 cases(41.1%) were myxoid type. 7. Immunohistochemically, the specimen of all tumors reacted positively to cytokeratin and vimentin marker.

• 치위생과학회지
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• 제23권4호
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• pp.369-377
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• 2023

Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

• Journal of Periodontal and Implant Science
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• 제31권1호
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• pp.109-122
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• 2001

Gingival fibroblasts(GF) and periodontal ligament fibroblasts(PDLF) are the major cellular components of periodontal soft connective tissues, but the precise molecular biological differences between these cells are not yet known. In the present study, we investigated the expression of S100A4, S100A2 calcium-binding protein and osteoblast-specific factor 2(OSF-2, Periostin) mRNA in GF and PDLF in vitro through the process of reverse transcription-polymerase chain reaction(RT-PCR) and Northern blot analysis in each. Human GF and PDLF were isolated from the gingival connective tissue and the middle third of freshly extracted healthy third molars. They were cultured in Dulbecco's Modified Eagle Medium(DMEM) containing 10% fetal bovine serum and cells in the third passage were used in the experiments. After extracting total RNA from cultured cells, RT-PCR and Northern analysis were performed using S100A4-, S100A2- and Periostin-specific oligonucleotide primers and subcloned cDNA probes in each. In PT-PCR and Northern analysis, the expression of S100A4 and Periostin mRNA in GF was slightly detectable. Interestingly, the expression of S100A4 and periostin mRNA in PDLF was much higher than that in GF. On the other hand, S100A2 mPNA was highly expressed in both GF and PDLF. Since there was a marked difference of S100A4 and Periostin expression between GF and PDLF in vitro, these data suggest that S100A4 and periostin could be used as a useful marker for distinguishing cultured gingival fibroblasts and periodontal ligament cells.

• Molecules and Cells
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• 제21권1호
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• pp.29-33
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• 2006

We have established three immortal bovine muscular epithelial (BME) cell lines, one spontaneously immortalized (BMES), the second SV40LT-mediated (BMEV) and the third hTERT-mediated (BMET). The morphology of the three immortal cell lines was similar to that of early passage primary BME cells. Each of the immortal cell lines made cytokeratin, a typical epithelial marker. BMET grew faster than the other immortal lines and the BME cells, in 10% FBS-DMEM medium, whereas neither the primary cells nor the three immortal cell lines grew in 0.5% FBS-DMEM. The primary BME cells and the immortal cell lines, with the exception of BMES, made increasing amounts of p53 protein when treated with doxorubicin, a DNA damaging agent. On the other hand, almost half of the cells in populations of the three immortal cell lines may lack $p16^{INK4a}$ regulatory function, compared to primary BME cells that were growth arrested by enforced expression of $p16^{INK4a}$. In soft-agar assays, the primary cells and immortal cell lines proved to be less transformed in phenotype than HeLa cells. The three immortal epithelial-type cell lines reported here are the first cell lines established from muscle tissue of bovine or other species.

• Applied Biological Chemistry
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• 제42권1호
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• pp.20-28
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• 1999

Biopolymer를 생산하는 Bacillus속의 균주개량의 목적으로 biopolymer를 생산하는 균주인 Bacillus subtilis K-1과 유당 이용능이 있는 Bacillus coagulans의 원형질체 형성과 재생에 관하여 조사하였다. 영양요구성과 항생제 내성의 marker가 부여된 두 변이주의 원형질체 형성조건에서 Bacillus subtilis mutant SM-2의 경우, 대수기 중기에 penicillin G(1.0 unit/ml)를 첨가한 다음 1.5시간 반응 후 삼투압 안정제로서 0.4 M sucrose와 $25\;{\mu}g/ml$의 lysozyme이 함유된 lysis fluid(LF, pH 7.0)내에서 $37^{\circ}$, 40분간 반응시켰을 때, 원형질체 형성율은 99.6%, 세포벽 재생율 2.4%였다. Bacillus coagulans mutant CM-12의 경우 대수기 중기에 penicillin G 0.3 unit/ml와 glycine 0.5%를 혼합첨가하고 1시간 반응시킨 후 삼투압 안정제로서 0.6 M lactose와 $300\;{\mu}g/ml$의 lysozyme이 함유된 LF(pH 7.0)내에서 $37^{\circ}$, 30분간 재생시켰을 때, 원형질체 형성율 90.8%, 세포벽 재생율 2.2%였다. 세포벽 재생효율을 높이기 위한 재생배지는 trypticase soy broth(TSB)에 0.4 M sucrose, 0.7% casamino acd, 1% PVP, 25 mM $CaCI_2$, 25 mM $MgCI_2$, 1.5% agar가 함유된 배지에 0.4% soft agar로서 중층 했을 때 Bacillus subtilis SM-2의 재생율은 5.1%, Bacillus coagulans CM-12의 재생율은 10.3%로 $2{\sim}4$배 가량 향상 되었다.