• Title/Summary/Keyword: Single Cell Surgery

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MicroRNA-497 Suppresses Proliferation and Induces Apoptosis in Prostate Cancer Cells

  • Wang, Li;Li, Bo;Li, Lei;Wang, Te
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3499-3502
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    • 2013
  • MicroRNAs (miRNAs) are a class of endogenously expressed small, non-coding, single-stranded RNAs that negatively regulate gene expression, mainly by binding to 3'- untranslated regions (3'UTR) of their target messenger RNAs (mRNAs), which cause blocks of translation and/or mRNA cleavage. Recently, miRNAprofiling studies demonstrated the microRNA-497 (miR-497) level to be down-regulated in all prostate carcinomas compared with BPH samples. The purpose of this study was to investigate the potential role of miR-497 in human prostate cancer. Proliferation, cell cycle and apoptosis assays were conducted to explore the potential function of miR-497 in human prostate cancer cells. Results showed that miR-497 suppressed cellular growth and initiated G0/G1 phase arrest of LNCaP and PC-3 cells. We also observed that miR-497 increased the percentage of apoptotic cells by increasing caspase-3/7 activity. Taken together, our results demonstrated that miR-497 can inhibit growth and induce apoptosis by caspase-3 activation in prostate cancer cells, which suggest its use as a potential therapeutic target in the future.

Abdominal Endometriosis Diagnosed by Fine Needle Aspiration Cytology -A Report of Two Cases- (세침흡인 세포검사로 진단된 복벽의 자궁내막증 -2예 보고-)

  • Chung, Myoung-Ja;Jeong, Yeon-Jun;Hwang, Ho-Myong;Jang, Kyu-Yoon;Moon, Woo-Sung;Kang, Myoung-Jae;Lee, Dong-Geun
    • The Korean Journal of Cytopathology
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    • v.15 no.1
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    • pp.70-73
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    • 2004
  • The incidence of endometriosis in post-operative abdominal scars is rare. We describe two cases of abdominal endometriosis diagnosed by fine needle aspiration (FNA). Both patients presented with subcutaneous masses at previous cesarean section scars with cyclic symptoms of pain. The cytologic smears were cellular and comprised two distinct cell populations consisting of epithelial and stromal components. An epithelial component consisted of flat sheets of polygonal cells and the second stromal component showed crowded clusters of spindle cells or isolated single cells. Hemosiderin-laden macrophages were found in the background. FNA offers a safe and effective tool for diagnosis of abdominal wall endometriosis.

The Excitatory Mechanism of Substance P in the Antral Circular Muscle of Guinea Pig Stomach

  • Jun, Jae-Yeoul;Kim, Sung-Joon;Choi, Youn-Baik;Kim, Ki-Whan
    • The Korean Journal of Physiology
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    • v.28 no.1
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    • pp.51-59
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    • 1994
  • This study was carried out to elucidate the excitatory mechanisms of Substance P in the antral circular muscle, using isometric contraction recording, conventional microelectrode method and whole-cell patch clamp technique. Substance P produced tonic and phasic contractions in a dose-dependent manner and depolarized membrane potential with increased amplitude of slow waves in muscle strips. Voltage-dependent $Ca^{2+}$ currents were increased by the application of Substance P from a holding potential of -60mV to 50mV in 10mV steps and this effect was blocked by the addition of an antagonist. Also Substance P increased transient and spontaneous oscillatory $K^+$ outward currents. The enhanced outward currents were abolished by apamin in dispersed single cells. These results suggest that the depolarization of membrane potential by Substance P activates voltage-dependent $Ca^{2+}$ channels, which represents an excitatory response in the antral circular muscle and led to an increase in $Ca^{2+}\;activated\;K^+\;currents$.

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Survival Following Non Surgical Treatments for Oral Cancer: a Single Institutional Result

  • Larizadeh, Mohammad Hasan;Shabani, Mohammad
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4133-4136
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    • 2012
  • Aim: To report the results of radiotherapy with or without chemotherapy in the patients with oral cancer. Methods: Over the 2003-2009 periods, a total number of 69 patients with squamous cell carcinoma of the oral cavity that refused surgery or had unresectable tumor were enrolled in this study. A total dose of 60 to 70 Gy (2 Gy per day) was given to the primary tumor and clinically positive nodes. In the patients with locoregionally advanced disease (57 patients with $T_3$, $T_4$ lesions and/ or $N^+$) induction chemotherapy following by concomitant chemoradiation was used. Induction chemotherapy consisted of 3 cycles of Cisplatin and 5-Flourouracil with or without Docetaxel. Weekly cisplatin was used in concomitant protocol. Kaplan-Meier method was used to calculate overall survival. Log-rank test and Cox regression model were used for comparison purposes. Results: Median follow-up was 32 months. The mean age of the patients was 59.2 years. The overall response rate after induction chemotherapy was 68.4%. Actuarial overall survival rates after 2 and 3 years were 38% and 26%, respectively. Clinical stage emerged as the only independent predictor of survival. Conclusion: Outcome of the patients with oral cancer is poor. Presenting with an advanced stage lesion contributed to this result. The role of chemotherapy in advanced cases remains to be defined.

A STUDY ON MUTATIONS OF P53 TUMOR SUPPRESSOR GENE IN ORAL TUMORS (구강종양에서 p53 종양억제 유전자의 돌연변이에 관한 연구)

  • Joo, Seong-Chai;Pyo, Sung-Woon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.1
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    • pp.45-52
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    • 2000
  • Nowadays, there are a lot of evidence that mutation of the p53 tumor suppressor gene is one of the most common genetic abnormalities in neoplastic progression. In this study, we analyzed 20 specimens of oral tumors(squamous cell carcinoma 14 cases, ameloblastoma 3 cases, adenoid cystic carcinoma 2 cases, malignant schwannoma 1 case)using polymerase chain reaction and direct sequencing which used an automated DNA sequencer and software for detection of mutations. Polymerase chain reactions were performed with 4 sets of primers encompassing exon 5, 6, 7, 8, and direct sequencing method was employed. The results were as followings. 1. We detected 10 point mutations out of 20 specimens (50%). 2. The genetic alterations included 7 mis-sense mutations resulting in single amino acid subtitutions, 2 silent mutations, 1 non-sense mutations encoding a stop codon. 3. Mutations were mostly in exon 7(7 out of 10 mutations, 70%) and involved codons 225, 234, 235, 236, 238, 247. 4. Therse were 4 cases of $T{\rightarrow}A$ transversion, 2 cases of $C{\rightarrow}A$ transversion, $A{\rightarrow}G$ transition, 1 case of $C{\rightarrow}G$, $T{\rightarrow}G$ transversion respectively. 5. We could find out point mutations more conveniently using PCR - Automated Direct Sequencing method.

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Antagonists of NMDA Receptor, Calcium Channel and Protein Kinase C Potentiate Inhibitory Action of Morphine on Responses of Rat Dorsal Horn Neuron

  • Shin, Hong-Kee;Kim, Yeon-Suk;Jun, Jong-Hun;Lee, Seo-Eun;Kim, Jae-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.5
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    • pp.251-254
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    • 2003
  • The present study was designed to examine whether the co-application of morphine with $Ca^{2+}$ channel antagonist $(Mn^{2+},\;verapamil)$, N-methyl-D-aspartate (NMDA) receptor antagonist (2-amino-5-phosphonopentanoic acid$[AP_5]$, $Mg^{2+}$) or protein kinase C inhibitor (H-7) causes the potentiation of morphine-induced antinociceptive action by using an in vivo electrophysiological technique. A single iontophoretic application of morphine or an antagonist alone induced weak inhibition of wide dynamic range (WDR) cell responses to iontophoretically applied NMDA and C-fiber stimulation. Although there was a little difference in the potentiating effects, the antinociceptive action of morphine was potentiated when morphine was iontophoretically applied together with $Mn^{2+}$, verapamil, $AP_5$, $Mg^{2+}$ or H-7. However, the potentiating action between morphine and each antagonist was not apparent, when the antinociceptive action evoked by morphine or the antagonist alone was too strong. These results suggest that the potentiating effect can be caused by the interaction between morphine and each antagonist in the spinal dorsal horn.

SARS-CoV-2 Infection of Airway Epithelial Cells

  • Gwanghui Ryu;Hyun-Woo Shin
    • IMMUNE NETWORK
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    • v.21 no.1
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    • pp.3.1-3.16
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    • 2021
  • Coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading worldwide since its outbreak in December 2019, and World Health Organization declared it as a pandemic on March 11, 2020. SARS-CoV-2 is highly contagious and is transmitted through airway epithelial cells as the first gateway. SARS-CoV-2 is detected by nasopharyngeal or oropharyngeal swab samples, and the viral load is significantly high in the upper respiratory tract. The host cellular receptors in airway epithelial cells, including angiotensin-converting enzyme 2 and transmembrane serine protease 2, have been identified by single-cell RNA sequencing or immunostaining. The expression levels of these molecules vary by type, function, and location of airway epithelial cells, such as ciliated cells, secretory cells, olfactory epithelial cells, and alveolar epithelial cells, as well as differ from host to host depending on age, sex, or comorbid diseases. Infected airway epithelial cells by SARS-CoV-2 in ex vivo experiments produce chemokines and cytokines to recruit inflammatory cells to target organs. Same as other viral infections, IFN signaling is a critical pathway for host defense. Various studies are underway to confirm the pathophysiological mechanisms of SARS-CoV-2 infection. Herein, we review cellular entry, host-viral interactions, immune responses to SARS-CoV-2 in airway epithelial cells. We also discuss therapeutic options related to epithelial immune reactions to SARS-CoV-2.

CHARACTERISTICS OF FIBROUS DYSPLASIA DERIVED CELLS (섬유성이형성증 유래세포의 특성연구)

  • Lee, Chan-Hee;Han, Ihn;Seo, Byoung-Moo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.5
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    • pp.304-309
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    • 2009
  • Purpose: Fibrous dysplasia (FD) is a fibro-osseous disease associated with activating missense mutations of the gene encoding the $\alpha$-subunit of stimulatory G protein. FD may affect a single bone (called monostotic form) or multiple bones (called polyostotic form). The extent of lesions reflects the onset time of mutation. In this study, cells from monostotic FD in maxilla of a patient were isolated and cultured in vitro for characterization. Materials and Methods: The single cells were released from FD lesion which was surgical specimen from 15 years-old boy. These isolated cells were cultured in vitro and tested their proliferation activity with MTT assay. In osteogenic media, these cells underwent differentiation process comparing with its normal counterpart i.e. bone marrow stromal cells. The proliferated FD cells were detached and transplanted into the dordsal pocket of nude mouse and harvested in 6 weeks and 12 weeks. Results and Summary: FD cells have an increased proliferation rate and poor differentiation. As a result, cells isolated from FD lesion decreased differentiation into osteoblast and increased proliferation capacity. MTT assay presented that proliferation rate of FD cells were higher than control. However, the mineral induction capacity of FD was lesser than that of control. Monostotic FD cells make fewer amounts of bone ossicles and most of them are woven bone rather than lamellar bone in vivo transplantation. In transplanted FD cells, hematopoietic marrow were not seen in the marrow space and filled with the organized fibrous tissue. Therefore, they were recapitulated to the original histological features of FD lesion. Collectively, these results indicated that the FD cells were shown that the increased proliferation and decreased differentiation potential. These in vitro and in vivo system can be useful to test FD cell's fate and possible.

Improvement of Cell Viability Using a Rho-associated Protein Kinase (ROCK) Inhibitor in Human Dental Papilla derived Single-induced Pluripotent Stem Cells (ROCK 억제제를 통한 사람 치유두 조직 유래 단일 사람 유도만능줄기세포의 생존성 향상)

  • Shim, Yoo-Jin;Kang, Young-Hoon;Kim, Hyeon-Ji;Kim, Mi-Jeong;Lee, Hyeon-Jeong;Son, Young-Bum;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.29 no.8
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    • pp.895-903
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    • 2019
  • The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.

Rab25 Deficiency Perturbs Epidermal Differentiation and Skin Barrier Function in Mice

  • Jeong, Haengdueng;Lim, Kyung-Min;Goldenring, James R.;Nam, Ki Taek
    • Biomolecules & Therapeutics
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    • v.27 no.6
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    • pp.553-561
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    • 2019
  • Rab25, a member of the Rab11 small GTPase family, is central to achieving cellular polarity in epithelial tissues. Rab25 is highly expressed in epithelial cells of various tissues including breast, vagina, cervix, the gastrointestinal tract, and skin. Rab25 plays key roles in tumorigenesis, mainly by regulating epithelial differentiation and proliferation. However, its role in skin physiology is relatively unknown. In this study, we demonstrated that Rab25 knock-out (KO) mice show a skin barrier dysfunction with high trans-epidermal water loss and low cutaneous hydration. To examine this observation, we investigated the histology and epidermal differentiation markers of the skin in Rab25 KO mice. Rab25 KO increased cell proliferation at the basal layer of epidermis, whereas the supra-basal layer remained unaffected. Ceramide, which is a critical lipid component for skin barrier function, was not altered by Rab25 KO in its distribution or amount, as determined by immunohistochemistry. Notably, levels of epidermal differentiation markers, including loricrin, involucrin, and keratins (5, 14, 1, and 10) increased prominently in Rab25 KO mice. In line with this, depletion of Rab25 with single hairpin RNA increased the expression of differentiation markers in a human keratinocyte cell line, HaCaT. Transcriptomic analysis of the skin revealed increased expression of genes associated with skin development, epidermal development, and keratinocyte differentiation in Rab25 KO mice. Collectively, these results suggested that Rab25 is involved in the regulation of epidermal differentiation and proliferation.