• KSBB Journal
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• v.18 no.1
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• pp.55-61
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• 2003

To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.

• Applied Microscopy
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• v.37 no.2
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• pp.83-91
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• 2007

The present study examined the epidermal changes of adhesive disks which occur during attachment in Parthenocissus tricuspidata using scanning and transmission electron microscopy. Several adhesive disks, each covered with a bract, develop from the shoot apical meristem during early development. In the initial stage, the adhesive disks are club-shaped and their upper and lower epidermis are indistinguishable. However, in the actively growing stage, they become spherical and both epidermis are clearly differentiated into the adventitious roots. Prior to wall attachment, the adhesive disks exhibit adaxial convex and abaxial concave shapes, and electron-dense substances are abundant in the vacuoles of epidermal cells. The peripheral area of the adhesive disk is adhered first to the wall surface, while the central area is drawn inward in a vacuum-like state during attachment. As the attachment progresses and the electron-dense substances continue to discharge, the upper and lower epidermis rapidly undergo deterioration and the disks shrink considerably. At this stage, structural changes of the lower epidermis occur much faster than in the upper one. The discharged substance is accumulated on the wall surface, and this aids the attachment of adhesive disks on the wall for long periods. In this manner, the shape and structure of the adhesive disk epidermis change drastically from initial growth to the mature stage. Further, the role of electron-dense substance and shrinkage of the disk during attachment has been discussed in Parthenocissus tricuspidata.

• Applied Microscopy
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• v.37 no.2
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• pp.93-102
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• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.4
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• pp.416-421
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• 2008

This study was performed to determine the antimutagenic and anticytotoxic effects of soybean paste (doenjang) added deep sea water salt and see tangle in Salmonella Typhimurium TA98, TA100 and human cancer cell lines. In the Ames test, methanol extract of doenjang did not exhibit any mutagenicity but showed substantial inhibitory effects against mutation induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1-oxide (4NQO). The methanol extracts of doenjang ($200{\mu}g$/plate) added deep sea salt and see tangle (doenjang C) showed approximately 89.1% and 70% inhibitory effect on the mutagenesis induced by MNNG and 4NQO against TA100 strain, whereas 84.4% inhibitions were observed on the mutagenesis induced by 4NQO against TA98 strain. The cytotoxic effects of doenjang methanol extracts against the cell lines with human cervical adenocarcinoma (HeLa), human hepatocellular carcinoma (Hep3B), human gastric carcinoma (AGS), human lung carcinoma (A549) and human breast adenocarcinoma (MCF-7) were inhibited with the increase of the extract concentration. The treatment of 1.0 mg/mL doenjang C of methanol extracts showed strong cytotoxicities of 71%, 74.4%, 66.2%, 77.3%, and 71.2% against HeLa, Hep3B, AGS, A549, and MCF-7, respectively. In contrast 1 mg/mL treatment of doenjang C methanol extracts had only $10{\sim}40%$ cytotoxicity on normal human embryonal kidney cell (293). Doenjang methanol extract inhibited significantly the tumor growth in mice injected sarcoma-180 cells. Especially, doenjang C methanol extract showed an inhibition of tumor cell activity of 33% by the administration of 25 mg/kg methanol extracts.

• Journal of Chest Surgery
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• v.23 no.3
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• pp.452-464
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• 1990

It is well known that extracellular Calcium plays a very important role in several steps of smooth muscle excitability and contractility, and there have been many concerns about factors influencing the distribution of extracellular Ca++ and the Ca++ flux through the cell membrane of the smooth muscle. Based on the assumption that Mg++ may also play an important role in the excitation and contraction processes of the smooth muscle by taking part in affecting Ca++ distribution and flux, many researches are being performed about the exact role of Mg++, especially in the vascular smooth muscle. But yet the effect of Mg++ in the smooth muscle activity is not clarified, and moreover the mechanism of Mg++ action is almost completely unknown. Present study attempted to clarify the effect of Mg++ on the excitability and contractility in the multiunit and unitary smooth muscle, and the mechanism concerned in it. The preparations used were the guinea-pig aortic strip as the experimental material of the multiunit smooth muscle and the rat uterine strip as the one of the unitary smooth muscle. The tissues were isolated from the sacrificed animal and were prepared for recording the isometric contraction. The effects of Mg++ and Ca++ were examined on the electrically driven or spontaneous contraction of the preparations. And the effects of these ions were also studied on the K+ or norepinephrine contracture. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% 02 and kept at 35oC. The results obtained were as follows: 1] Mg++ suppressed the phasic contraction induced by electrical field stimulation dose-dependently in the guinea-pig aortic strip, while the high concentration of Ca++ never recovered the decreased tension. These phenomena were not changed by the a - or b - adrenergic blocker. 2]Mg++ played the suppressing effect on the low concentration [20 and 40 mM] of K+-contracture in the aortic muscle, but the effect was not shown in the case of 100mM K+-contracture. 3] Mg++ also suppressed the contracture induced by norepinephrine in the aortic preparation. And the effect of Mg++ was most prominent in the contracture by the lowest [10 mM] concentration of norepinephrine. 4] In both the spontaneous and electrically driven contractions of the uterine strip, Mg++ decreased the amplitude of peak tension, and by the high concentration of Ca++ the amplitude of tension was recovered unlike the aortic muscle. 5] The frequency of the uterine spontaneous contraction increased as the [Ca++] / [Mg++] ratio increased up to 2, but the frequency decreased above this level. 6] Mg++ decreased the tension of the low[20 and 40mM] K+-contracture in the uterine smooth muscle, but the effect did not appear in the 100mM K+-contracture. From the above results, the following conclusion could be made. 1] Mg++ seems to suppress the contractility directly by acting on the smooth muscle itself, besides through the indirect action on the nerve terminal, in both the aortic and uterine smooth muscles. 2] The fact that the depressant effect of Mg++ on the K+-contracture is in inverse proportion to an increase of K+ concentration appears resulted from the extent of the opening state of the Ca++ channel. 3] Mg++ may play a depressant role on both the potential dependent and the receptor-operated Ca++ channels. 4] The relationship between the actions of Mg++ and Ca++ seems to be competitive in uterine muscle and non-competitive in aortic strip.

• Journal of Chest Surgery
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• v.40 no.2 s.271
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• pp.114-121
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• 2007

Background: Apoptosis plays a crucial role in carcinogenesis, as well as in development and tissue homeostasis. Terminal deoxyribonucleotidyl transferase mediated neck end labelling (TUNEL) and in situ nick end labelling (ISEL) have been used to investigate the apoptosis in tissues. Since the introduction of the M30 monoclonal antibody to overcome drawbacks of TUNEL and ISEL, the apoptosis in various tumors, with the exception of pulmonary carcinomas, has been studied. In this study, attempts were made to examine the correlation of apoptosis in non-small cell carcinomas, using both M30 and the expression of p53 protein, with the clinicopathological factors. Material and Method: Forty five patients with surgically resected non-small cell carcinomas were included. Immunohistochemical staining with M30 and p53 monoclonal antibody were peformed, and their expressions compared with the clinicopathological features. The overall survival time and recurrence-free survival time were calculated, and the factors influencing the survival time analyzed using a univariate analysis. The effects of the expression stati of M30 and p53 on the risks of cancer related to both death and recurrence were evaluated using a multivariate analysis. Result: The p53 positive group had many more M30 positive cells than the p53 negative group (p53 positive group; $61.7{\pm}26.8$ cells vs. p53 negative group; $45.6{\pm}29.6$ cells, p=0.005) and significantly more p53 positive patients showing at least 10 positive cells (apoptotic index, $Al{\ge}1$) on M30 staining (p53 positive group; 52.4% (11/21) vs. p53 negative group 16,7% (4/24), p=0.025). In the univariate analysis, the survival times in relation to smoking (pack-year), performance status (PS) and Al showed significant differences. The multivariate analysis demonstrated the relative risk (R.R) of cancer death increased almost 7.5-fold (R.R 7.482; 95% Cl $1.886{\sim}29.678$; p=0.004) and the risk of recurrence almost 3,8-fold (R.R 3.795; 95% Cl: $1.184{\sim}12.158$; p=0.025) in the high Al (${\ge}1$) compared to the low Al (<1) group. There was no prognostic effect of p53 expression on the survival time or risk of cancer death and recurrence. Conclusion: In non-small cell lung carcinomas, M30 immunohistochemistry was an excellent method for analyzing apoptosis; the high apoptotic index could be an adverse prognostic predictive factor.

• Radiation Oncology Journal
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• v.13 no.3
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• pp.259-266
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• 1995

Purpose : The aim of this study is to analyze the survival rate, treatment failure and complication of radiation therapy alone in stage III uterine cervical cancer. Materials and Methods : From January 1980 through December 1985, 227 patients with stage III uterine cervical cancer treated with radiation therapy at Kosin Medical Center were retrospectively studied. Among 227 patients, 72 patients($317{\%}$) were stage IIIa, and 155 patients($68.3{\%}$) were stage IIIb according to FIGO classification. Age distribution was 32-71 years (median: 62 years). Sixty nine patients($95.8{\%}$) in stage IIIa and 150 patient ($96.8{\%}$) in stage IIIb were squamous cell carcinoma. pelvic lymph node metastasis at initial diagnosis was 8 patients($11.1{\%}$) in stage IIIa and 29 patients($18.7{\%}$) in stage IIIb, Among 72 patients with stage IIIa, 36 patients ($50{\%}$) were treated with external radiation therapy alone by conventional technique (180-200 cGy/fr.) and 36 patients($50{\%}$) were treated with external radiation therapy with intracavitary radiotherapy(ICR) with $Cs^{137}$ sources, and among 155 patients with stage IIIb, 80 patients ($51.6{\%}$) were treated with external radiation therapy alone and 75 patients ($48.4{\%}$) were treated with external radiation therapy with ICR. Total radiation doses of stage IIIa and IIIb were 65-105 Gy(median: 78.5 Gy) and 65-125.5 Gy (median 83.5 Gy). Survival rate was calculated by life-table method. Results : Complete response rates were $58.3{\%}$(42 patients) in stage IIIa and $56.1{\%}$(87 patients) in stage IIIb. Overall 5 year survival rates were $57{\%}$ in stage IIIa and $40{\%}$ in stage IIIb. Five year survival rates by radiation technique in stage IIIa and IIIb were $64{\%},\;40{\%}$ in the group treated in combination of external radiation and ICR, and $50\%,\;40\%$ in the group of external radiation therapy alone(P=NS). Five year survival rates by response of radiation therapy in stage IIIa and IIIb were $90\%,\;66\%$ in responder group and $10\%,\;7\%$ in non-responder group (P<0.001) There were statistically no significant differences of 5 year survival rate by total radiation doses and external radiation doses(40 Gy vs 50 Gy) of whole or true pelvis in stage IIIa and IIIb(P=NS). Treatment failures rates were $40.3\%$(29 patients) in stage IIla and $57.4\%$(89 patients) in stage IIIb. 17 patients ($23.6\%$) in stage IIIa and 46 patients ($29.7\%$) in stage IIIb experienced complications. Total radiation doses more than 85 Gy produced serious complication in both stage IIIa($50\%$) and IIIb($50\%$). Serious complication rates were higher in group received external radiation doses of 50 Gy than 40 Gy to whole or true pelvis in stage IIIa and IIIb. Serious rectal complication developed in rectal doses more than 65 Gy, and serious bladder complication developed in bladder doses more than 75 Gy. Major cause of death was cachexia due to locoregional failure in both stage IIIa($34.7\%$) and IIIb($43.9\%$). Conclusion : From this study, we found that external radiation therapy with ICR was found to have a tendency to be superior to external radiation therapy alone in survival rate, local control rate and complication rate but not different in statistics, and external radiation doses of 50 Gy than 40 Gy to whole or true pelvis produced serious rectal and bladder complications in stage III uterine cervical cancer.

• Parasites, Hosts and Diseases
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• v.29 no.3
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• pp.293-310
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• 1991

Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

• Parasites, Hosts and Diseases
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• v.25 no.1
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• pp.69-82
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• 1987

The prevalences of the cuke belonging to genus Metagonimus hove been reported along the upper stream of inhabitants by several workers since 1980, however the taxonomical problems of the fluke was not yet settled. The larval flukes; cercaria and metacercaria as well as their intermediate hosts, and adult were studied in order to identify the Mepagonimus in the areas. The results obtained are summarized as follows: 1. The snails, Semisulcospira globus were collected (rom the three different localities along the upper stream of the River. The cercariae were found from 125(7.2%) out of 1,730 snails by natural emerging method, and were identified into 5 species including Metagenimus sp. (3.7%), Pseudexorchis major(1.4%), Cercaria nipponensis (0.9%), Cercaria incerpa(0.6%), and Cercaria yoshidae(0.6%). Cercariae of Metagonimus species had four to dye oral spines on its anterior of the first line. 2. The cercariae of Metagonimus were experimentally exposed to goldfish. nfection rate was 22.9% out of 105 goldfish, and the encysted metacercariae were found in fins(86.7%) and on scales (13.7%) of the fishes, but not in their muscle, head or visceral organs. 3. Seven species of ask were caught in the Daecheong Reservoir and the upper stream. Infestations with metacercaria of Metagonimus were found 100% in Opsariichtys widens and the parasitized numbers of the metacercariae were observed from 250 to 2,400 per fish. In the upper stream, Zacco termmincki, Z. platypus and Pseudogobio esocinus were infected 100% with the metacercaria, on the other hand, the fishes caught in the reservoir showed the lower infestation rates, and a few metacercariae found in the fishes Carassius carassius and Cyprinus carpio in the reservoir and the stream. The majority of metacercariae was detected only on the scales of fishes. 4. In order to know the infectivity and the distribution patterns in the intestine of hosts, rats and dogs were infected with the metacercariae obtained from O. bidens and Z. platypus. In addition the metacercariae obtained from Z. temmincki, P. esocinus and goldfish were given to the rats. The recovery rates of the worms in the small intestine of dogs were higher (63.3~65.8%) than those of the rats (3.5~31.6%). The flukes were found mostly in the middle and the lower part of small intestine of the rats and the dogs, but no worm was collected in the upper part of the intestine of rats. 5. The sixte of adult flukes varied by the hosts. In the adult cukes, oral sucker was smaller than ventral sucker, and the right and left testes were located diagonally, the uterine tubules circled around the upper left testis. The average egg sixte was $29.1{\times}1.7{\mu\textrm{m}}$. According to the above results, the cukes belonging to genus Metagonimus distributed along the Geum River was concluded to be identical with Miyata type of M. yokogawai as that Saito had proposed.

• Parasites, Hosts and Diseases
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• v.29 no.3
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• pp.245-258
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• 1991

In order to determine the antigenic localization in the tissues of the adult Metagonimus yokegawai, immunogoldlabeling method was applied using serum immunoglobulins (IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size: 12 nm) , It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.