Earticle

Ovulation resembles a tissue remodeling process such as a blood coagulation. The present study was aimed to examine the involvement of tissue factor, a primary factor for extrinsic coagulation pathway, in the ovulation. Northern blot analysis revealed that mRNA levels of tissue factor and tissue factor pathway inhibitor 2 (TFPI-2) in the ovary were stimulated by human chorionic gonadotropin (hCG) treatment in surperovulation model, of immature rats. Real-time PCR analysis demonstrated that the expression of tissue factor and TFPI-2 was stimulated in granulosa and theca cells of preovulatory follicles, respectively. The induction of tissue factor mRNA was blocked by the progesterone receptor antagonist RU486. Tissue factor protein was not detected in the ovary by Western blot and immunohistochemical analysis due to the lack of a specific antibody. Interestingly, the levels of tissue factor and TFPI-2 mRNA were increased in the ovarian cells of rats induced ovarian hyperstimulation syndrome (OHSS) and in granulosa cells of OHSS patients undergiong in vitro fertilization. The present findings indicate the stimulation of tissue factor system during ovulation, and in OHSS patients, implicating the possible involvement of tissue factor system in OHSS.

Ovulation resembles a tissue remodeling process such as a blood coagulation. The present study was aimed to examine the involvement of tissue factor, a primary factor for extrinsic coagulation pathway, in the ovulation. Northern blot analysis revealed that mRNA levels of tissue factor and tissue factor pathway inhibitor 2 (TFPI-2) in the ovary were stimulated by human chorionic gonadotropin (hCG) treatment in surperovulation model, of immature rats. Real-time PCR analysis demonstrated that the expression of tissue factor and TFPI-2 was stimulated in granulosa and theca cells of preovulatory follicles, respectively. The induction of tissue factor mRNA was blocked by the progesterone receptor antagonist RU486. Tissue factor protein was not detected in the ovary by Western blot and immunohistochemical analysis due to the lack of a specific antibody. Interestingly, the levels of tissue factor and TFPI-2 mRNA were increased in the ovarian cells of rats induced ovarian hyperstimulation syndrome (OHSS) and in granulosa cells of OHSS patients undergiong in vitro fertilization. The present findings indicate the stimulation of tissue factor system during ovulation, and in OHSS patients, implicating the possible involvement of tissue factor system in OHSS.

Background: A pressure ulcer is common in soft tissue over the greater trochanter (GT) in side-lying position, and sustained tissue deformation induced by the prolonged external force is a primary cause, which can be discussed with soft tissues' viscoelastic properties (i.e., stress relaxation, creep response). Objects: Using an automated hand-held indentation device, we measured the viscoelastic properties of soft tissue over the hip area, in order to examine how the properties are affected by site with respect to the GT. Methods: Twenty participants (15 males and 5 females) who aged from 21 to 32 were participated. An automated hand-held indentation device was used to measure the stress relaxation time and creep response. Trials were acquired for three different locations with respect to the GT (i.e., right over the GT, 6 cm anterior or posterior to the GT). For each location, five trials were acquired and averaged for data analyses. Results: Soft tissues' stress relaxation time and creep response were associated with site (F = 23.98, p < 0.005; F = 24.09, p < 0.005; respectively). The stress relaxation time was greatest at posterior gluteal region (19.22 ± 2.49 ms), and followed by anterior region (15.39 ± 2.47 ms) and right over the GT (14.40 ± 3.18 ms). Similarly, creep response was greatest at posterior gluteal region (1.16 ± 0.14), and followed by anterior region (0.95 ± 0.14) and right over the GT (0.89 ± 0.18). Conclusion: Our results showed that the stress relaxation and creep were greatest at the posterior gluteal region and least at right over the GT, indicating that the gluteal soft tissue is more protective to the prolonged external force, when compared to the trochanteric soft tissue. The results suggest that a risk of pressure ulcer over the GT may decrease with slightly posteriorly rotated side-lying position.

The present study was performed to identify the role of plasminogen activator (PA) and the location of PA expression in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from ovary in pre-ovulatory (Pre-Ov), post-ovulatory stage (Post-Ov) and early to mid-luteal stage (Early-mid L). The uterus tissue was immediately fixed by PBS with 10% formalin. There were fixed porcine uterus tissue for 24 hours at room temperature and porcine uterus tissue dehydrate for 12 hour in sucrose solution. For immunohistochemical staining, porcine uterus tissues were cut to 4 μm by micro frozen section microtome. The nucleus and cytoplasm of porcine uterus tissues were stained by Hematoxin and Eosin. Porcine uterus tissues were evaluated by Immunofluorescence using anti-tissue type PA (tPA) and urokinase type PA (uPA). The location of PA expression was identified by observing the PA fluorescence using fluorescent microscope and optical telescopes. As a results, when Pre-Ov and Post-Ov were identified endometrial blood vessel in an inner layer that were observed tPA and uPA. Especially, expression of PA was observed around secretory gland. But the expression of PA were not confirm in Early-mid L. Also, The expression of PA were higher in Post-Ov than Early-mid L. In conclusion, during the estrous cycle, the expression of PA were increased from Pre-Ov to Post-Ov and was decreased from Post-Ov to Early-mid L.

The present study was performed to identify the role of plasminogen activator (PA) and the location of PA expression in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from ovary in pre-ovulatory (Pre-Ov), post-ovulatory stage (Post-Ov) and early to mid-luteal stage (Early-mid L). The uterus tissue was immediately fixed by PBS with 10% formalin. There were fixed porcine uterus tissue for 24 hours at room temperature and porcine uterus tissue dehydrate for 12 hour in sucrose solution. For immunohistochemical staining, porcine uterus tissues were cut to 4 μm by micro frozen section microtome. The nucleus and cytoplasm of porcine uterus tissues were stained by Hematoxin and Eosin. Porcine uterus tissues were evaluated by Immunofluorescence using anti-tissue type PA (tPA) and urokinase type PA (uPA). The location of PA expression was identified by observing the PA fluorescence using fluorescent microscope and optical telescopes. As a results, when Pre-Ov and Post-Ov were identified endometrial blood vessel in an inner layer that were observed tPA and uPA. Especially, expression of PA was observed around secretory gland. But the expression of PA were not confirm in Early-mid L. Also, The expression of PA were higher in Post-Ov than Early-mid L. In conclusion, during the estrous cycle, the expression of PA were increased from Pre-Ov to Post-Ov and was decreased from Post-Ov to Early-mid L.

This study was performed to investigate the effect of ultrasound irradiation on epidermal growth factor(EGF) expression in rat wound tissue. Skin wounds were created below 5mm to both sides of scapular inferior angle. The right wound was used experimental side and left was used control side. Ultrasound was irradiated pulse rate $20\%$, frequency 1MHz, intensity $0.5W/cm^2$ for 5 minutes during 3days. After sonication during 3 days, rats were sacrified. The expression of epidermal growth factor evaluated immunohistochemistry on mouse anti-EGF. In the control side, a little expression of EGF was observed at epidermis and dermis. In the experimental side, A strong immunostaining was seen at epidermis and dermis. This study suggests that ultrasound irradiation is effective on EGF expression in wound tissue.

Recently, with the increase of meat production, high quality and safety of meat have been strongly emphasized by Korean consumers. Marbling in beef has been regarded as an important criterion deciding meat quality in Korea. The purpose of this study was to identify the transcriptional level of insulin-like growth factor-1 (IGF-1) in longissimus muscle samples of 46 Hanwoo. The level of IGF-1 transcripts was measured by real-time polymerase chain reaction (PCR) and molecular connection of IGF-1 was analyzed using the Pathway Studio program (Ver 9.0). Increase of marbling score (MS) induced increase of IGF-1 transcripts level in the muscle and there is a significant correlation (p<0.05) between IGF-1 mRNA expression and MS. The pathway study showed that IGF-1 genes are regulated in insulin, fatty acid synthase, leptin, and corticotrophin releasing hormone. These results suggest that IGF-1 might be used as a useful marker for the improvement of economic traits in Hanwoo.

Insulin-like growth factor-I (IGF-I) 유전자의 발현은 사람 및 쥐에서 두 개의 promoters (P1과 P2)로부터의 전사와 alternative RNA splicing 및 differential RNA polyadenylation과 같은 복잡한 기전들에 의하여 조절되는데 조직에 따라 성장호르몬을 포함한 여러 요소들이 관여하는 것으로 알려져있다. 또한 사람의 IGF-I 유전자 exon 1의 upstream에 존재하는 P1에 hepatocyte nuclear factor 1α와 CAAT/enhancerbinding protein (C/EBP) isoform 들이 결합하여 조직 및 발달단계 특이한 발현에 중요한 역할을 할 것으로 제안되었지만, exon 1의 downstream sequence가 IGF-I 유전자의 조직 특이 적 발현을 조절하는 지에 대하여는 연구되어 있지 않다. 연령이 다른 쥐의 간 및 뇌 조직에서 total RNA를 분리하고 solution hybridization/RNase protection 방법으로 분석하여 IGF-I 유전자의 발현이 태어난 후 간 조직에서는 점차적으로 증가하였지만 뇌조직에서는 감소하여 발달단계에 따라 조직특이하게 발현되는 것을 확인하였다. IGF-I exon 1의 주요한 전사 개시점으로부터 아래쪽에 존재하는 C/EBP 결합부위를 포함하고 있는 cis-acting element에 해당하는 oligonucleotide들과 간 및 뇌조직에서 분리한 핵단백질들을 이용하여 DNA-결합 활성을 가진 분자량이 다른 C/EBPα나 C/EBPβ 단백질들을 확인하였으며 southwestern 및 western immnoblotting 분 석을 하여 간 조직의 핵 추출물에서는 42C/EBPα, 와 p38C/EBPα, p35C/EBPα, p38C/EBPβ, 그리고 p35C/EBPβ가 IGF-I exon 1 oligonucleotide와 복합체를 형성하고 뇌 조직에서는 p42C/EBPα과 p38C/EBPβ 가 복합체 형성에 관여하는 것으로 나타났다. 이러한 결과들은 FRE-C/EBP isoform 복합체 형성이 IGF-I 유전자 발현의 조직 특이적 조절에 중요한 역할을 할 것으로 제안한다.

The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBPα and C/EBPβ proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42C/EBPα, p38C/EBPα, p35C/EBPα, p38C/EBPβ, and p35C/EBPβ form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42C/EBPα and p38C/EBPβ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.

배경: Tissue factor inhibitor (TFPI)는 응고작용을 조절하는 물질로, 출혈성 질환의 치료를 위하여 TFPI를 억제함으로써 지혈 효과를 얻고자 하는 연구가 진행되고 있다. 이 연구에서는 정상 성인군과 출혈 위험이 높은 것으로 알려진 질환군의 혈액 TFPI 농도를 비교하고, 나아가 혈액 TFPI 농도 및 활성도가 응고작용에 미치는 영향을 조사하였다. 방법: 정상 성인군, 만성 신장질환군, 만성 간질환군, 혈우병군의 혈액 TFPI를 측정하여 비교하였고, TFPI 농도 및 활성도가 응고작용에 미치는 영향을 분석하기 위해 thrombin generation assay (TGA)를 시행하였다. 결과: 정상 성인군, 만성 간질환군, 만성 신장질환군, 혈우병군의평균 혈액 TFPI 농도는 각각 16.47 ng/mL, 24.90 ng/mL, 49.47 ng/mL, 17.35 ng/mL이었다. 정상 성인군과 만성 간질환군의 경우 혈액 TFPI 농도와 TGA parameter 사이에 상관관계를 보였다(P<0.05). 만성 간질환군과 혈우병군의 TGA parameter 측정값은 정상 성인군과 의미 있는 차이를 보였다(P<0.05). 항 TFPI 항체를 첨가한 FVIII de¬cient plasma에서 항 TFPI 항체의 농도에 따라 TGA parameter가 정상화되는 경향을 보였다. 하지만 FVIII 농도에 따른 TGA parameter의 차이는 뚜렷하지 않았다. 결론: 혈액 TFPI 농도는 질환군의 출혈경향과 어느 정도 연관성을 보이지만 응고과정에 영향을 끼치는 요인이 다양하기 때문에 혈액 TFPI 농도의 영향을 독립적으로 해석하기 어려울 것으로 보인다. 하지만 FVIII 부족으로 인한 출혈증상에 대한 치료법 개발을 위해서 TFPI를 연구할 가치가 있다.

Background: Tissue factor pathway inhibitor (TFPI) regulates blood coagulation. To treat hemorrhagic disease, studies have been conducted to achieve a hemostatic effect by inhibiting TFPI. In this study, blood TFPI levels of healthy and hemorrhagic disease groups were compared, and the effects of blood TFPI level and its activity on coagulation were investigated. Methods: The blood TFPI levels of the healthy, chronic liver disease (CLD), chronic kidney disease (CKD), and hemophilia groups were measured and compared. Thrombin generation assay (TGA) was performed to investigate the effects of TFPI level and its activity on coagulation. Results: The mean blood TFPI levels of the healthy, CLD, CKD, and hemophilia groups were 16.47 ng/mL, 24.90 ng/mL, 49.47 ng/mL, and 17.35 ng/mL, respectively. The healthy and CLD groups showed correlations between their blood TFPI levels and TGA parameters (P <0.05). The TGA parameters of the CLD and hemophilia groups showed significant differences compared with those of the healthy group (P <0.05). In FVIII-deficient plasma with anti-TFPI antibodies, the TGA parameters tended to be normalized according to anti-TFPI antibody levels. However, the differences in TGA parameters caused by FVIII concentration were unclear. Conclusions: Although the TFPI level was related to bleeding in the disease groups, it is difficult to interpret its effects independently because of the various factors that affect the coagulation process. However, TFPI is worth studying to facilitate the development of a treatment strategy for bleeding due to lack of FVIII

Tissue factor (TF, tissue thromboplastin) is a membrane bound glycoprotein, which acceler-ates the blood clotting, activating both the intrinsic and the extrinsic pathways to serve as a cofactor for activated factor VII (Vila). The TF-factor Vila complex (TF/VIIa) proteolytically activates factors IX and X, which leads to the generation of thrombin and fibrin clots. In order to isolate TF inhibitors, by means of a bioassay-directed chromatographic separation technique, from the leaves of Eriobotrya japonica Lindley (Rosaceae), a known sesquiterpene glycoside (2) and ferulic acid (3) were isolated as inhibitors that were evaluated using a single-clotting assay method for determining TF activity. Another sesquiterpene glycoside (1) was also isolated but was inactive in the assay system. Compound 3 was yielded by alkaline hydrolysis of compound 2. The structures of compounds 1, 2, and 3 were identified by means of spectral analysis as $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-a-L-rhamnopyranosyl-(1{\rightarrow}2)-[{\alph}-L-rhamnopyrano-syl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl nerolidol$ (1), $3-O-{\alph}-L-rhamnopyranosyl-(1{\rightarrow}4)-{\alph}-L-rhamnopyr-anosyl-(1{\rightarrow}2)-[{\alph}-L-(4-trans-feruloyl)-rhamnopyranosyl-(1{\rightarrow}6)]-{\beta}-D-glucopyranosyl$ nerolidol (2) and ferulic acid (3), respectively. Compounds 2 and 3 inhibited 50% of the TF activity at con-centrations of 2 and $369{\;}\mu\textrm{m}/TF$ units, respectively.

Tissue factor (TF, tissue thromboplastin or coagulation factor III) accelerates the blood clotting, activating both the intrinsic and the extrinsic pathways to serve as a cofactor. In order to isolate TF inhibitors from the fruits of Chaenomeles sinensis, an activity-guided purification utilizing a bio-assay method of prothrombin time prolongation, was carried out to yield five active flavoniods such as hovetrichoside C (1) ($IC_{50}$ = 14.0 $\mu$g), luteolin-7-Ο-$\beta$-D-glucuronide (3) ($IC_{50}$ = 31.9$\mu$g), hyperin (4) ($IC_{50}$ = 20.8 $\mu$g), avicularin (6) ($IC_{50}$ = 54.8 $\mu$g) and quercitrin (10) ($IC_{50}$ = 135.7 $\mu$g), along with other inactive compounds such as ($\pm$)-(2E,4E)-Ο-$\beta$-D-glucopyranosyl-4'-hydroxy-$\beta$-ionylideneacetic acid ester (2), genistein-7-Ο-$\beta$-D-glucopyranoside (5), luteolin-3'-methoxy-4'-Ο-$\beta$-D-glucopyranoside (7), luteolin-7-Ο-$\beta$-D-glucuronide methyl ester (8), tricetin-3'-methoxy-4'-Ο-$\beta$-D-glucopyranoside (selagin-4'-Ο-$\beta$-D-glucopyranoside) (9), (-)-epicatechin (11), luteolin-4'-Ο-$\beta$-D-glucopyranoside (12) and apigenin-7-Ο-$\beta$-D-glucuronide methyl ester (13). The structures of the isolated compounds were elucidated through spectral analysis. Among them, compounds 1 to 9, 12 and 13 were isolated for the first time from the fruits of this plant and the compound 9 is a new flavonoid.

Associations between periodontal infection and cardiovascular disease have been documented. Porphyromonas gingivalis is a well-established periodontal pathogen, and tissue factor (TF) is a key initiator of the coagulation cascade. In this context, P. gingivalis has been reported to enhance TF expression in human endothelial cells. The present study investigated the underlying mechanisms of TF induction by P. gingivalis in human umbilical vein endothelial cells. P. gingivalis increased TF expression in a dose- and time-dependent manner. Not only live bacteria but also glutaraldehyde-fixed bacteria increased TF expression to the same extent. However, sonicates of P. gingivalis did not induce TF expression. Cytochalasin D and SMIFH2, which are inhibitors of actin polymerization and actin nucleation, respectively, inhibited the TF expression induced by P. gingivalis. Finally, TF production was decreased or increased in the presence of various signaling inhibitors, including mitogen-activated protein kinases. These results suggest that P. gingivalis induces endothelial TF expression by a bacterial internalization-dependent mechanism and through diverse signal transduction mechanisms.

Objective: This study aimed to explore the expression of tissue factor (TF), protease activated receptor-2 (PAR-2), and matrix metalloproteinase-9 (MMP-9) in the MCF-7 breast cancer cell line and influence on invasiveness. Methods: Stable MCF-7 cells transfected with TF cDNA and with TF ShRNA were established. TF, PAR-2, and MMP-9 protein expression was analyzed using indirect immunofluorescence and invasiveness was evaluated using a cell invasion test. Effects of an exogenous PAR-2 agonist were also examined. Results: TF protein expression significantly differed between the TF cDNA and TF ShRNA groups. MMP-9 protein expression was significantly correlated with TF protein expression, but PAR-2 protein expression was unaffected. The PAR-2 agonist significantly enhanced MMP-9 expression and slightly increased TF and PAR-2 expression in the TF ShRNA group, but did not significantly affect protein expression in MCF-7 cells transfected with TF cDNA. TF and MMP-9 expression was positively correlated with the invasiveness of tumor cells. Conclusion: TF, PAR-2, and MMP-9 affect invasiveness of MCF-7 cells. TF may increase MMP-9 expression by activating PAR-2.

Objective: Tissue factor (TF) is expressed abnormally in certain types of tumor cells, closely related to invasion and metastasis. The aim of this study was to construct a human gastric cancer cell line SGC7901 stably-transfected with human TF, and observe effects on oxaliplatin-dependent inhibition of invasion and the apoptosis induction. Methods: The target gene TF was obtained from human placenta by nested PCR and introduced into the human gastric cell line SGC7901 through transfection mediated by lipofectamine. Stably-transfected cells were screened using G418. Examples successfully transfected with TF-pcDNA3 recombinant (experimental group), and empty vector pcDNA3 (control group) were incubated with oxaliplatin. Transwell chambers were used to show change in invasive ability. Caspase-3 activity was detected using a colorimetric method and annexin-V/PI double-staining was applied to detect apoptosis. Results: We generated the human gastric cancer cell line SGC7901/TF successfully, expressing TF stably and efficiently. Compared with the control group, invasion increased, whereas caspase-3 activity and apoptosis rate were decreased in the experimental group. Conclusion: TF can enhance the invasive capacity of gastric cancer cells in vitro. Its increased expression may reduce invasion inhibition and apoptosis-inducing effects of oxaliplatin and therefore may warrant targeting for improved chemotherapy.

Tissue factor (TF), a principal initiator of the veertebrate coagulation cascade, is expressed in organ tissues, cells and blood. TF is konwn to be induced in endothelial cells, monocytes and macrophages by inflammatory stimuli and in many pathologic conditions. By using the modified method for in vido TF activity assay, we found that turpentine oil injection as an inflamatory stimulus also induced the TF activity in lung and brain tissues of rats. And the age-related increase in Tf activity was observed in healthy rat brain tissue.

Associations between periodontal infection and cardiovascular disease have been documented. Porphyromonas gingivalis is a well-established periodontal pathogen, and tissue factor (TF) is a key initiator of the coagulation cascade. In this context, P. gingivalis has been reported to enhance TF expression in human endothelial cells. The present study investigated the underlying mechanisms of TF induction by P. gingivalis in human umbilical vein endothelial cells. P. gingivalis increased TF expression in a dose- and time-dependent manner. Not only live bacteria but also glutaraldehyde-fixed bacteria increased TF expression to the same extent. However, sonicates of P. gingivalis did not induce TF expression. Cytochalasin D and SMIFH2, which are inhibitors of actin polymerization and actin nucleation, respectively, inhibited the TF expression induced by P. gingivalis . Finally, TF production was decreased or increased in the presence of various signaling inhibitors, including mitogen-activated protein kinases. These results suggest that P. gingivalis induces endothelial TF expression by a bacterial internalization-dependent mechanism and through diverse signal transduction mechanisms. solutions. Tertiary dentin is formed when odontoblasts are directly affected by various stimuli. Recent preclinical studies have reported that stimulation of the Wnt/β-catenin signaling pathway could facilitate the formation of reparative dentin and thereby aid in the structural and functional development of the tertiary dentin. A range of signaling pathways, including the Wnt/β-catenin pathway, is activated when dental tissues are damaged and the pulp is exposed. The application of small molecules for dentin regeneration has been suggested as a drug repositioning approach. This study reviews the role of Wnt signaling in tooth formation, particularly dentin formation and dentin regeneration. In addition, the application of the drug repositioning strategy to facilitate the development of new drugs for dentin regeneration has been discussed in this study.

Molecular imaging with targeted contrast agents enables tissues to be distinguished by detecting specific cell-surface receptors. In the present study, a ligand-targeted acoustic nanoparticle system is used to identify angioplasty-induced expression of tissue factor by smooth muscle cell within carotid arteries. Pig carotid arteries were overstretched with balloon catheters, treated with tissue factor-targeted or a control nanoparticle system, and imaged with intravascular ultrasound before and after treatment. Tissue factor-targeted emulsion bound and increased the echogenicity and gray-scale levels of overstretched smooth muscle cell within the tunica media, versus no change in contralateral control arteries. Expression of stretch-induced tissue factor in carotid artery media was confirmed by immunohistochemistry. The potential for abnormal thrombogenicity of balloon-injured arteries, as reflected by smooth muscle expression of tissue factor, was imaged using a novel, targeted, nanoparticulate ultrasonic contrast agent.