Background: With the development of the molecular biological methods, studies of the early diagnosis of lung cancer and the detection in the preneoplastic state by using genetic probes in the high risk groups are widely investigated. In lung cancer, squamous cell carcinoma is considered to progress from the normal bronchial mucosa to the preneoplastic state, and finally to the invasive carcinoma. In this study, we investigated the expression of p53 and c-erbB2 in the normal bronchi and the cancer tissues in patients with squamous cell lung cancer to evaluate the possibility of using these immunohistochemical markers as the diagnostic and prognostic parameters of patients with squamous cell lung cancer. Method: The normal and cancerous bronchial tissues of 25 patients with squamous cell carcinoma of the lung, surgically resected from May 1995 to November 1996, were immunohistochemically stained with the monoclonal antibodies to p53(DAKO-p53) and c-erbB2(phamingen 15821A) respectively. We compared the expression status of these markers between the normal bronchial mucosa and the tumor tissue, and also investigated the relationship between the expression status of these markers in tumor tissues and the pathological stage, and the survival time. Results: The pathological stage was as follows; stage I, II were found in 5 patients respectively, stage IIIA was in 8 patients, stage IIIB was in 4 patients, and stage IV was in 3 patients. The expression rate of p53 in the squamous cell lung cancer was 48%, and it was not expressed in the normal bronchial mucosa. The expression status was increased as the pathological stage advanced(p=0.0091 by test of trend). But there were no relationship between the expression of p53 and the median survival time. C-erbB2 did not yield a significantly meaningful result. Conclusion: p53 was not found in the normal bronchial mucosa, but it was expressed in 48% of the tumor tissue. And the expression rate increased as the pathological stage advanced. So it would be helpful to apply the immunochistochemical stain with p53 in the bronchial biopsy specimen in the early diagnosis trial or staging of squamous cell lung cancer.
This study was carried out to evaluate the effect of different freezing and thawing rates on the viability, motility and acrosomal changes of frozen canine spermatozoa. The ejaculated semen was extended with Tris-egg yolk buffer containing 8% glycerol and equilibrated for 60 min after cooled to 4$^{\circ}C$ for 58 min. The straws were cryopreserved gradually by slow-cooling at different distance(6, 10 and 17 cm, respectively) from the liquid nitrogen (L$N_2$) to achieve temperature rate of 3, 8.9 and 19$^{\circ}C$ /min. Thawing of the straws was performed in a water bath fur 2 min at 37$^{\circ}C$ and 55$^{\circ}C$ , respectively. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Concentration of the ejaculated fresh semen was normal range of 3.44 $\times$ 10$^{8}$ /ml. Freezing temperature were reduced to -110, -70 and -35$^{\circ}C$, as higher distance from liquid nitrogen, 6, 10 and 17 cm, respectively. Freezing at 3$^{\circ}C$/min in distance of 17 cm from liquid nitrogen yielded better motility, viability and rate of intact acrosome than 8.9 or 19$^{\circ}C$/min and the optimal thawing was 37$^{\circ}C$ for 2 min.
Park, Ki Ho;Kang, Seok Yong;Kang, Anna;Jung, Hyo Won;Park, Yong-Ki
The Korea Journal of Herbology
/
v.34
no.6
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pp.79-89
/
2019
Objective : This study investigated the anti-diabetic effects of DM1, a herbal mixture with Atractylodis Rhizoma, Anemarrhenae Rhizoma, and Cinnamomi Cortex in high fat diet (HFD)-induced diabetic mice and the mechanism in C2C12 mouse skeletal muscle cells. Methods : The C57B/6 mice were fed high fat for 12 weeks, and then administrated DM1 extract (500 mg/kg, p.o.) for 4 weeks. The changes of body weight, calorie and water intakes, fasting blood glucose levels and the serum levels of glucose, insulin, triglyceride, HDL-cholesterol, AST and ALT were measured in mice. The histological changes of liver and pancreas tissues were also observed by H&E stain. C2C12 myoblasts were differentiated into myotubes and then treated with DM1 extract (0.5, 1, and 2 mg/㎖) for 24 hr. The expression of myosin heavy chain (MHC), PGC1α, Sirt1 and NRF1, and the AMPK phosphorylation were determined in the myotubes by western blot, respectively. Results : The DM1 extract administration significantly decreased the calorie and water intakes, glucose, triglyceride, AST and ALT levels and increased insulin and HDL-cholesterol in HFD-induced diabetic mice. DM1 extract inhibited lipid accumulation in liver tissue and improved glucose tolerance. In C2C12 myotubes, DM1 treatment increased the expression of MHC, PGC1α, Sirt-1, NRF-1 and the AMPK phosphorylation. Conclusion : In our results indicate that DM1 can improve diabetic symptoms by decreasing the obesity, glucose tolerance and fatty liver in HFD-induced diabetic mice, and responsible mechanism is might be related with energy enhancement.
Embryonic genome activation (EGA) is the first major transition that occurs after fertilization, and entails a dramatic reprogramming of gene expression that is essential for continued development. Although it has been suggested that EGA in porcine embryos starts at the four-cell stage, recent evidence indicates that EGA may commence even earlier; however, the molecular details of EGA remain incompletely understood. The RNA polymerase II of eukaryotes transcribes mRNAs and most small nuclear RNAs. The largest subunit of RNA polymerase II can become phosphorylated in the C-terminal domain. The unphosphorylated form of the RNA polymerase II largest subunit C-terminal domain (IIa) plays a role in initiation of transcription, and the phosphorylated form (IIo) is required for transcriptional elongation and mRNA splicing. In the present study, we explored the nuclear translocation, nuclear localization, and phosphorylation dynamics of the RNA polymerase II C-terminal domain in immature pig oocytes, mature oocytes, two-, four-, and eight-cell embryos, and the morula and blastocyst. To this end, we used antibodies specific for the IIa and IIo forms of RNA polymerase II to stain the proteins. Unphosphorylated RNA polymerase II stained strongly in the nuclei of germinal vesicle oocytes, whereas the phosphorylated form of the enzyme was confined to the chromatin of prophase I oocytes. After fertilization, both unphosphorylated and phosphorylated RNA polymerase II began to accumulate in the nuclei of early stage one-cell embryos, and this pattern was maintained through to the blastocyst stage. The results suggest that both porcine oocytes and early embryos are transcriptionally competent, and that transcription of embryonic genes during the first three cell cycles parallels expression of phosphorylated RNA polymerase II.
The objective of this study is to provide the information on the small-scale model mix proportion when the behavior of prototype concrete pavement is studied through small-scale model experiments. However it is difficult to obtain a model material to simulate the prototype concrete by scaling the individual components according to the laws of similitude. In this paper, the stress-strain behavior in uniaxial compression is used as a means to correlate material similitude between the prototype and the model concrete. Based on the results of experiments, we compared the stress-strain curves of prototype and model concrete mixes using a nondimensional basis. In order to simulate the stress-stain curves of prototype concrete, it is important that various mix proportions of model concrete selected properly which are varied from aggregate grading, cement-aggregate and sand-aggregate ratio.
The aim of study was to elucidate dyeability and antimicrobial and antifungal activity of silk fabrics dyed with Sophora Radix extracts according to different mordants. Dyes were extracted from Sophora Radix using ethanol. Then, silk fabrics were dyed with extracts two times by post-mordanting method in which the extract was 60%(owf), the mordant was 3%(owf), L.R was 1:20, the temperature was $60~60^\circ{C}$, the time of dyeing was 60min., and the time of mordanting was 60min. The dyeability was evaluated by surface color, K/S values and durability of dye. The skin microorganisms used in this study was S. sureus, B. subtilis, S. epidermidis, P. acnes, P. aeruginosa, E coli, A. niger, C. albicans and T. mentatrophytes. The results are as follows; 1. When mordants were added, K/S value of silk dyed was not improved much and surface color was 2.2Y to 8.8Y in H(hue) value which indicated greenish yellow to raddish yellow 2. The color fastness tests to light, perspiration, dry-cleaning, rubbing, and stain fabric washing show 4~5th degree which were valuated excellent. The color fastness to fade washing was improved to 3~4th degree by addition of $K_2CrO_7$ mordants. 3. Antibacterial activity of silk dyed using no-mordant as well as mordants was excellent on S. aureus, B. subtilis, S.epidermidis and P.acnes, but showed poor antibacterial activities on P.aeruginosa and E.coli such as gram negative baterials 4. Antifungal activity of silk dyed with ethanol extracts was good on A.niger, C.candida and T.mentagrophytes. Especially, on T. mentagrophytes there was no growth of fungus during 72 gous in silk dyed mordanting with $SnCl_2\cdot{2H}_2O$.
BACKGROUND/OBJECTIVES: Several medicinal properties of Smilax china L. have been studied including antioxidant, anti-inflammatory, and anti-cancer effects. However, the antiobesity activity and mechanism by which the water-soluble fraction of this plant mediates its effects are not clear. In the present study, we investigated the lipolytic actions of the water-soluble fraction of Smilax china L. leaf ethanol extract (wsSCLE) in 3T3-L1 adipocytes. MATERIALS/METHODS: The wsSCLE was identified by measuring the total polyphenol and flavonoid content. The wsSCLE was evaluated for its effects on cell viability, lipid accumulation, glycerol, and cyclic adenosine monophosphate (cAMP) contents. In addition, western blot analysis was used to evaluate the effects on protein kinase A (PKA), PKA substrates (PKAs), and hormone-sensitive lipase (HSL). For the lipid accumulation assay, 3T3-L1 adipocytes were treated with different doses of wsSCLE for 9 days starting 2 days post-confluence. In other cell experiments, mature 3T3-L1 adipocytes were treated for 24 h with wsSCLE. RESULTS: Results showed that treatment with wsSCLE at 0.05, 0.1, and 0.25 mg/mL had no effect on cell morphology and viability. Without evidence of toxicity, wsSCLE treatment decreased lipid accumulation compared with the untreated adipocyte controls as shown by the lower absorbance of Oil Red O stain. The wsSCLE significantly induced glycerol release and cAMP production in mature 3T3-L1 cells. Furthermore, protein levels of phosphorylated PKA, PKAs, and HSL significantly increased following wsSCLE treatment. CONCLUSION: These results demonstrate that the potential antiobesity activity of wsSCLE is at least in part due to the stimulation of cAMP-PKA-HSL signaling. In addition, the wsSCLE-stimulated lipolysis induced by the signaling is mediated via activation of the ${\beta}$-adrenergic receptor.
Background: Microsatellites are short-tandem repeated uncleotide sequences present throughout the human genome. Alterations of microsatellites have been termed microsatellite instability(MI). It has been generally known that microsatellite instability detected in hereditary non-polyposis colorectal cancer (HNPCC) reflects genetic instability that is caused by impairments of DNA mismatch repair system regarding as a novel tumorigenic mechanism. A number of studies reported that MI occurred at varying frequencies in non-small cell lung carcinoma (NSCLC). However It has been unproven whether MI could be a useful market of genetic instability and have a clinical significance in NSCLC. Material and Method : We have examined whether MI can be observed in thirty NCSLC using polymerase chain reaction whether such alterations are associated with other molecular changes such as p53, K-ras and c-myc oncoproteins expression detected by immunohistochemical stain,. Result: MI(+) was observed in 16.6%(5/30) and MI(-) was 83.3% (25/30) Average age was 50$\pm$7.5 year-old in MI(+) group and 57$\pm$6.6 year-old in MI(-) group. Two year survival rate in MI(=) group (20% 1/5) was worse than MI(-) group (64% 16/25) with a statistic difference. (P=0.04) The positive rate of K-ras oncoprotein expression and simultaneous expression of 2 or 3 oncoproteins expression were higher in MI(+) group than MI(-) group with a statistic difference(P=0.05, P=0.01) Conclusion: From, these results the authors can conclude that MI is found in some NSCLC and it may be a novel tumorigenic mechanism in some NSCLC. We also conclude that MI could be used as another poor prognostic factor in NSCLS.
This study was carried out to investigate the immunopathological effects of 7,12-Dimethylbenz[a]anthracene(DMBA) on spleen in mice. DMBA was administered subcutaneously to BALB/C mice by interscapular single injection of 50 or 100${\mu}g/g$ of body weight. Each DMBA treatment group and additional corn oil control group of mice were studied on day 1,3,7,14 and 21 following the injection of DMBA. DMBA treatment resulted in marked decrease in weights and cellularity of spleen. Spleen weights showed the greatest decrease at 14days after 50${\mu}g/g$ DMBA treatment, and at 21days after 100${\mu}g/g$ DMBA treatment. Spleen cellularity was similarly deceased in comparison with spleen weights. Spleen showed morphologically no typical changes throughout the experiment after 50${\mu}g/g$ DMBA treatment. Following the treatment of 100${\mu}g/g$ DMBA the spleen showed severe fibrosis, hemosiderin precipitation, and megakaryocytes decrease in red pulp at 14 days, while hemopoietic function was partly restored in addition to the appearance of a few megakaryocytes at 21 days. In spleen sections treated with antibodies to IgM or Thy1.2, lymphocytes strongly stained with IgM antibody were infiltrated around the central artery within the white pulp, and T-lymphocytes of periarterial lymphatic sheath (PALS) were diminished and destructed in sections treated with Thy1.2 antibody, at 14 days after the treatment of 100${\mu}g/g$ DMBA. By the electron microscopy phagocytic epithelial cells or macrophages were remarkably increased in spleen at 14and 21days following the treatment of 100${\mu}g/g$ DMBA.
Park, Hong-Gyu;Kim, Jin-Moon;Kim, Jung-Mogg;Chung, Won-Yoon;Yoo, Yun-Jung;Cha, Jeong-Heon
Food Science and Biotechnology
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v.17
no.6
/
pp.1327-1331
/
2008
In order to determine whether peach contains compounds to regulate adipocyte differentiation, extracts of flesh/pericarp of yellow/white peach were prepared in water, ethyl acetate (EtOAc), or n-butanol solvent and determined for effects on adipocyte differentiation in C3H10T1/2 or 3T3-L1 cells. Interestingly, none of peach extracts has statistically significant stimulatory effect on the adipocyte differentiation in C3H10T1/2. Furthermore, the presence of EtOAc extract of white peach pericarp (WPP) was found to inhibit lipid accumulation in 3T3-L1 cells both by microscopic examination of Oil Red O-stained lipid droplets and by spectrophotometric quantification of extracted stain, indicating a significant inhibitory effect on adipocyte differentiation. The inhibition of lipid accumulation was accompanied by a significant decrease in the expression levels of adipocyte molecular markers-peroxisome proliferator-activated receptor $\gamma$, CAAT enhancer binding protein $\alpha$, and fatty acid-binding protein. Thus, this study determined that WPP EtOAc extract contains the inhibitory compound(s) on adipogenesis.
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