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Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

Applicability of plate tectonics to the post-late Cretaceous igneous activities and mineralization in the southern part of South Korea( I ) (한국남부(韓國南部)의 백악기말(白堊紀末) 이후(以後)의 화성활동(火成活動)과 광화작용(鑛化作用)에 대(對)한 판구조론(板構造論)의 적용성(適用性) 연구(硏究)( I ))

  • Min, Kyung Duck;Kim, Ok Joon;Yun, Suckew;Lee, Dai Sung;Joo, Sung Whan
    • Economic and Environmental Geology
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    • v.15 no.3
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    • pp.123-154
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    • 1982
  • Petrochemical, K-Ar dating, Sand Rb/Sr isotopes, metallogenic zoning, paleomagnetic and geotectonic studies of the Gyongsang basin were carried out to examine applicability of plate tectonics to the post-late Cretaceous igneous activity and metallogeny in the southeastern part of Korean Peninsula. The results obtained are as follows: 1. Bulgugsa granitic rocks range from granite to adamellite, whose Q-Ab-Or triangular diagram indicates that the depth and pressure at which the magma consolidated increase from coast to inland varying from 6 km, 0.5-3.3 kb in the coastal area to 17 km, 0.5-10 kb in the inland area. 2. The volcanic rocks in Gyongsang basin range from andesitic to basaltic rocks, and the basaltic rocks are generally tholeiitic in the coastal area and alkali basalt in the inland area. 3. The volcanic rocks of the area have the initial ratio of Sr^{87}/Sr^{86} varying from 0.706 to 0.707 which suggests a continental origin; the ratio of Rb/Sr changing from 0.079-0.157 in the coastal area to 0.021-0.034 in the inland area suggests that the volcanism is getting younger toward coastal side, which may indicate a retreat in stage of differentiation if they were derived from a same magma. The K_2O/SiO_2 (60%) increases from about 1.0 in the coastal area to about 3.0 in the inland area, which may suggest an increase indepth of the Benioff zone, if existed, toward inland side. 4. The K-Ar ages of volcanic rocks were measured to be 79.4 m.y. near Daegu, and 61.7 m.y. near Busan indicating a southeastward decrease in age. The ages of plutonic rocks also decrease toward the same direction with 73 m.y. near Daegu, and 58 m.y. near Busan, so that the volcanism predated the plutonism by 6 m.y. in the continental interior and 4 m.y. along the coast. Such igneous activities provide a positive evidence for an applicability of plate tectonics to this area. 5. Sulfur isotope analyses of sulfide minerals from 8 mines revealed that these deposits were genetically connected with the spacially associated ingeous rocks showing relatively narrow range of ${\delta}^{34}S$ values (-0.9‰ to +7.5‰ except for +13.3 from Mulgum Mine). A sequence of metallogenic zones from the coast to the inland is delineated to be in the order of Fe-Cu zone, Cu-Pb-Zn zone, and W-Mo zone. A few porphyry type copper deposits are found in the Fe-Cu zone. These two facts enable the sequence to be comparable with that of Andean type in South America. 6. The VGP's of Cretaceous and post Cretaceous rocks from Korea are located near the ones($71^{\circ}N$, $180^{\circ}E$ and $90^{\circ}N$, $110^{\circ}E$) obtained from continents of northern hemisphere. This suggests that the Korean peninsula has been stable tectonically since Cretaceous, belonging to the Eurasian continent. 7. Different polar wandering path between Korean peninsula and Japanese islands delineates that there has been some relative movement between them. 8. The variational feature of declination of NRM toward northwestern inland side from southeastern extremity of Korean peninsula suggests that the age of rocks becomes older toward inland side. 9. The geological structure(mainly faults) and trends of lineaments interpreted from the Landsat imagery reveal that NNE-, NWW- and NEE-trends are predominant in the decreasing order of intensity. 10. The NNE-trending structures were originated by tensional and/or compressional forces, the directions of which were parallel and perpendicular respectively to the subduction boundary of the Kula plate during about 90 m.y. B.P. The NWW-trending structures were originated as shear fractures by the same compressional forces. The NEE-trending structures are considered to be priginated as tension fractures parallel to the subduction boundary of the Kula plate during about 70 m.y. B.P. when Japanese islands had drifted toward southeast leaving the Sea of Japan behind. It was clearly demonstrated by many authors that the drifting of Japanese islands was accompanied with a rotational movement of a clock-wise direction, so that it is inferred that subduction boundary had changed from NNE- to NEE-direction. A number of facts and features mentioned above provide a suite of positive evidences enabling application of plate tectonics to the late Cretaceous-early Tertiary igneous activity and metallogeny in the area. Synthesizing these facts, an arc-trench system of continental margin-type is adopted by reconstructing paleogeographic models for the evolution of Korean peninsula and Japan islands. The models involve an extention mechanism behind the are(proto-Japan), by which proto-Japan as of northeastern continuation of Gyongsang zone has been drifted rotationally toward southeast. The zone of igneous activity has also been migrated from the inland in late-Cretaceous to the peninsula margin and southwestern Japan in Tertiary.

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The Effect of the Pain on the TMJ and Masticatory Muscles to Tension-type Headache (측두하악관절과 저작근의 통증이 긴장성 두통에 미치는 영향)

  • Kim, Jin-Suk;Auh, Q-Schick;Lee, Jin-Yong;Hong, Jung-Pyo;Chun, Yang-Hyun
    • Journal of Oral Medicine and Pain
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    • v.31 no.4
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    • pp.327-335
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    • 2006
  • Generally, Tension-Type Headache(TTH) patients exhibit muscle pain, but can also have TMJ pain, which includes mouth opening limitation or joint sounds. The purpose of our study is to observe the clinical pain characteristics between TTH patients with muscle pain and TMJ pain. One hundred sixty-seven patients were diagnosed with TTH according to the questionnaires based on the International Headache Society's proposal on the diagnostic criteria of TTH. The patients were classified into three group; arthralgia group (18 patients), myalgia group (50 patients) and arthromyalgia group (99 patients). TTH patients with pericranial muscle pain were classified in the myalgia group. TTH patients with temporal region pain were classified in the arthralgia group. TTH patients with both types of pain were classified in the arthromyalgia group. The parameters in the diagnostic criteria such as quality, intensity, laterality of pain, and aggravation due to physical activities were compared among the three groups. 1. There were no significant differences in the quality of pain among the three groups. 2. There were no significant differences in the intensity of pain among the three groups. 3. There were no significant differences in the laterality of pain among the three groups. 4. A higher percentage of patients in the arthromyalgia group experienced headaches that were aggravated due to physical activity (p=0.03) compared to the other groups. The results of this study show that TTH patients with both arthralgia (TMJ pain) and myalgia (pericranial muscle pain) are more aggravated by physical activity than TTH patients with either one.

Changes of Leaf Area Index, Physiological Activities and Soil Water in Tricholoma matsutake Producing Pine Forest Ecosystem (송이산 소나무림 생태계에서 엽면적지수와 생리적활동 및 토양수분의 변화)

  • Koo, Chang-Duck;Ka, Kang-Hyun;Park, Won-Chul;Park, Hyun;Ryu, Sung-Ryul;Park, Yong-Woo;Kim, Tae-Heon
    • Journal of Korean Society of Forest Science
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    • v.96 no.4
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    • pp.438-447
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    • 2007
  • The purpose of this study was to understand the ecosystem structure and function and soil water changes in Tricholoma matsutake producing pine stands. The investigated stands were pine forest in Sogrisan National Park in Chungbuk-do of Korea. For the purpose we investigated main vegetation, leaf area index(LAI) as ecosystem structural factors and measured photosynthesis, transpiration, xylem water potential, and soil water changes as ecosystem functional factors. Vertical vegetation structure of the site was composed of Pinus densiflora as a overstory species, Quercus mongolica as midstory, Rhododendron mucronulatum, R. schlippenbachii and Fraxinus sieboldiana as understory ones. In the stands LAI was 3.8 during June to September, 2.6 in October and 2.1 during November to April. Photosyntheses of the trees were 6.0 to $7.0{\mu}mol\;CO_2/m^2/s$ in August, and for P. densiflora about $4.0{\mu}mol\;CO_2/m^2/s$ and for Q. mongolica $2.0{\mu}mol\;CO_2/m^2/s$ in mid October. However, R. mucronulatum stopped fixing $CO_2$ and F. sieboldiana shed off the leaves already in mid October. Transpirations were 2.5 to $3.5mmol\;H_2O/m^2/s$ in late August and about $1.0mmol/H_2O/m^2/s$ in mid October. Plant water potentials were -10 to -22 bars for P. densiflora and -5 to -12 bars for the other woody species. The lowest potentials was in late August and highest in late October. Soil water in the stand was closely related to topography. Soil water contents were 7 to 11% at the ridge, 8 to 15% at the hillside and 11 to 19% at the base. Soil temperatures were 0.2 to $0.4^{\circ}C$ higher in T. matustuake colony than noncolony. Mid September soil temperature decreased to $19^{\circ}C$ at which T. matsutake forms primordia. In T. matsutake colony soil moisture was 0.5 to 2.0% lower due to metabolism for consuming water. We suggest that the complicate relationships between ecosystem structure and function in Tricholoma matsutake producing pine stand need to be further investigated.

Petrochemical Study on the Cretaceous Volcanic Rocks in Kageo island, Korea (가거도(소흑산도)의 백악기 화산암류에 대한 암석화학적 연구)

  • 김진섭;백맹언;성종규
    • The Journal of the Petrological Society of Korea
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    • v.6 no.1
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    • pp.19-33
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    • 1997
  • This study reports the results about the petrography and geochemical characteristics of 10 representative volacanic rocks. The Cretaceous volcanic rocks distributed in the vicinity of the Kageo island composed of andesitic rocks, dacitic welded tuff, and rhyolitic rocks in ascending order. Sedimentary rock is the basement in the study area covered with volcanic rocks. Andesitic rocks composed of pyroclastic volcanic breccia, lithic lapilli tuff and cryptocrystallin lava-flow. Most dacitic rocks are lapilli ash-flow welded tuff. Rhyolitic rocks consists of rhyolite tuff and rhyolite lava flow. Rhyolite tuff are lithic crystal ash-flow tuff and crystal vitric ash-flow tuff with somewhat accidental fragments of andesitic rocks, but dacitic rocks. The variation of major and trace element of the volcanic rocks show that contents of $Al_2O_3$, FeO, CaO, MgO, $TiO_2$ decrease with increasing of $SiO_2$. On the basis of Variation diagrams such as $Al_2O_3$ vs. CaO, Th/Yb vs. Ta/Yb, and $Ce_N/YB_N$ vs. $Ce_N$, these rocks represent mainly differentiation trend of calc-alkaline rock series. On the discriminant diagrams such as Ba/La and La/Th ratio, Rb vs. Y + Nb, the volcanic rocks in study area belongs to high-K Orogenic suites, with abundances of trace element and ternary diagram of K, Na, Ca. According to the tectonic discriminant diagram by Wood, these rocks falls into the diestructructive continental margin. K-Ar ages of whole rocks are from andesite to rhyolite $97.0{\pm}6.8~94.5{\pm}6.6,\68.9{\pm}4.8,\61.5{\pm}4.9~60.7{\pm}4.2$ Ma, repectively. Volcanic rocks in study area show well correlation to the Yucheon Group in terms of rock age dating and geochemcial data, and derived from andesitic calc-alkaline magma that undergone low pressure fractional crystallization dominated plagioclase at <30km.

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Association between frequency of convenience foods use at convenience stores and dietary quality among high school students in Incheon (인천지역 일부 고등학생의 편의점 편의식 이용빈도와 식사의 질과의 관련성)

  • Kim, Eun-Mi;Choi, Mi-Kyeong;Kim, Mi-Hyun
    • Journal of Nutrition and Health
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    • v.52 no.4
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    • pp.383-398
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    • 2019
  • Purpose: This study investigated an association between dietary quality and use of convenience foods at convenience stores among high school students. Methods: A total of 474 high school students (225 boys and 249 girls) residing in Incheon participated in this questionnaire survey in June 2018. The subjects were divided into three groups according to the frequency of consumption of convenience foods at convenience stores; less than once a week, 1 ~ 2 times a week, and more than 3 times a week. Dietary quality was assessed using a nutrient quotient for adolescents (NA-Q). Logistic regression was used to investigate an association between dietary quality and use of convenience foods at convenience stores among high school students. Results: For boys and girls, higher monthly allowance was significantly associated with the higher frequency of consumption of convenience foods at convenience stores, whereas school grade, mother's occupational status, family size, extracurricular study, and eating speed were not significantly associated with the frequency of consumption of convenience foods. Higher intake frequency of cookies or sweet and greasy bread, processed beverage, Ramyon, night-time snack, and street food was significantly associated with the higher frequency of consumption of convenience foods for boys or girls. Boys and girls, who had a higher frequency of consumption of convenience foods at convenience stores had significantly greater odds for being in the low grade of dietary quality, especially in the moderation factor. Conclusion: The students who used convenience stores more often appeared to have more monthly allowance and to consume undesirable foods more often. Higher frequency of using convenience foods at convenience stores among high school students was associated with lower dietary quality. These study results can support efforts to provide nutrition education programs and guidelines to students who frequently use convenience foods at convenience stores.

The aesthetics of irony in repetition and the difference of Oh! Soojung (<오! 수정>의 아이러니 미학 - 반복과 차이의 구조를 중심으로)

  • Suh, MyungSoo
    • 기호학연구
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    • no.57
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    • pp.121-153
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    • 2018
  • In terms of the story told, we see that Oh! Soojung(Virgin Stripped Bare by Her Bachelors) is a film of the ideololgy of masculinity. However, from the point of view of the manner of presenting story, Oh! Soojung is a film that aims to devalue this ideology. How will it be possible? This is the principle of the irony that the speaker, by saying P, wants to make Q listen that devalues and contradicts P. Our study is tempted to explain the process of interpreting the irony in the film. The ideology of the film occurs when the presupposed contents have become the subject. For example Cendrion who tells a story of a girl married to a prince presupposes that the girl, Cendrion, is obedient. The subject of this story is that the presupposition: /the girls who want to be happy must be obedient/, which represents the ideology of masculinity. Presupposed content thus imposes on the public a collective and conservative value, as its enunciator belongs to the collective voice. Since ironisation occurs when the utterance itself is annulled, one must also deny or cancel the story told of Oh! Soojng: /Jeahun who is rich and Soojung who is obedient and virgin have become lovers/. Since there is no semantic mark within the utterance, irony is a voice that comes from without; this is how we understand irony in a purely pragmatic way. The outer voices are two things: the way to build the story: question of focusing, ocularization and auricularization, and the way to present the story: question the order, the frequency or the plot. Our study is focused on the question of frequency at Oh! Soojung which has a repetition structure in which the memory of Jeahun and that of Soojung are represented one after the other. Since the memories of two characters are not identical, the repetition is accompanied by differences. The differences at first allow the public to build their own story from the di?g?se of the film and then make the audience fall into confusion where we can not be certain of what we see and know in the di?g?se of the film, and finally make their knowledge questionable. About repetition, so that it can have validity in terms of the informativeness of the utterance, it must deny the existence of the previous repetition. This is how repetition cancels itself and consequently the utterance. We see that the irony of Oh! Soojung occurs by repetition with differences that cancels the story of the film.

Anti-inflammatory Effects of Rumohra adiantiformis Extracts Fermented with Bovista plumbea Mycelium in LPS-stimulated RAW 264.7 Cells (LPS로 자극된 RAW 264.7 세포에서 찹쌀떡버섯 균사체로 생물전환된 루모라고사리 추출물의 항염증 효과)

  • Ji-Hye Hong;Eun-Seo Jang;Myung-Chul Gil;Gye Won Lee;Young Ho Cho
    • Journal of Life Science
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    • v.33 no.6
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    • pp.471-480
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    • 2023
  • This study was designed to evaluate the anti-inflammatory effects of Rumohra adiantiformis extracts fermented with Bovista plumbea mycelium (B-RAE) in LPS-stimulated RAW 264.7 cells. The total polyphenol and total flavonoid content of B-RAE were 379.26±7.77 mg/g and 50.85±3.08 mg/g, respectively. The results of measuring the antioxidant activity of B-RAE showed that it scavenges 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and superoxide anion radical in a dose-dependent manner. B-RAE inhibited nitric oxide (NO) production in a dose-dependent manner without affecting cell viability. The gene expression of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-lβ (IL-1β), and IL-6 was measured using real time quantitative reverse transcription PCR (qRT-PCR). We found that, compared to the LPS-treated group, B-RAE significantly reduced the mRNA levels of the pro-inflammatory cytokines in a concentration-dependent manner. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the phosphorylation of transcription factors such as nuclear factor-κB (NF-κB), and the mitogen-activated protein kinase (MAPK) signaling pathway proteins were assessed using Western blot analysis. We found that B-RAE significantly suppressed the expression of iNOS and COX-2, but their expression was increased by LPS treatment. In addition, the phosphorylation of NF-κB and IκB, which was increased by LPS treatment, was reduced with B-RAE treatment. The effect of B-RAE on the phosphorylation of the MAPK signaling pathway proteins was measured, and the phosphorylation of extracellular signal-regulated kinase (ERK) and the p38 MAPK proteins decreased in a dose-dependent manner, while the phosphorylation of c-Jun N-terminal kinase (JNK) increased. These anti-inflammatory effects of B-RAE may thus have been achieved through the high antioxidant activity, the inhibition of NO production through the suppression of iNOS and COX-2 expression, the inhibition of the NF-κB pathway, and the suppression of pro-inflammatory cytokine expression.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Diagnostic Performance of HPV E6/E7 mRNA and HPV DNA Assays for the Detection and Screening of Oncogenic Human Papillomavirus Infection among Woman with Cervical Lesions in China

  • Wang, Hye-young;Lee, Dongsup;Park, Sunyoung;Kim, Geehyuk;Kim, Sunghyun;Han, Lin;Yubo, Ren;Li, Yingxue;Park, Kwang Hwa;Lee, Hyeyoung
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7633-7640
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    • 2015
  • Background: Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and it is responsible for most cases of cervical uterine cancer. Although HPV infections of the cervix do not always progress to cancer, 90% of cervical cancer cases have been found to be associated with high risk HPV (HR-HPV) infection. HPV DNA testing is widely used, along with Papanicolaou (Pap) testing, to screen for cervical abnormalities. However, there are no data on the prevalence of genotype-specific HPV infections assessed by measuring HPV E6/E7 mRNA in women representative of the Chinese population across a broad age range. Materials and Methods: In the present study, we compared the results with the CervicGen HPV RT-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis. We also investigated the prevalence and genotype distribution of HPV infection with a total of 324 liquid-based cytology samples collected in western Shandong province, East China. Results: The overall HPV prevalences determined by HPV DNA and HPV E6/E7 mRNA assays in this study were 79.9% (259/324) and 55.6% (180/324), respectively. Although the positivity of HPV E6/E7 mRNA expression was significantly lower than HPV DNA positivity, the HPV E6/E7 mRNA assay showed greater specificity than the HPV DNA assay (88.6% vs. 48.1%) in normal cytology samples. The prevalence of Alpha-9 (HPV 16, 31, 33, 35, 52, and 58) HPV infection among these women accounted for up to 80.3% and 76.1% of the high-grade lesions detected in the HPV mRNA and DNA tests, respectively. The HR-HPV genotype distribution, based on HPV DNA and E6/E7 mRNA expression by age group in patients with cytologically confirmed lesions, was highest in women aged 40 to 49 years (35.9% for cytologically confirmed cases, Pearson correlation r value=0.993, p<0.001) for high-grade lesions. Among the oncogenic HR-HPV genotypes for all age groups, there was little difference in the distribution of HPV genotypes between the HPV DNA (HPV -16, 53, 18, 58, and 33) and HPV E6/E7 mRNA (HPV -16, 53, 33, 58, and 18) assays. HPV 16 was the most common HPV genotype among women with high-grade lesions. Conclusions: Our results suggest that the HPV E6/E7 mRNA assay can be a sensitive and specific tool for the screening and investigation of cervical cancer. Furthermore, it may provide useful information regarding the necessity for early cervical cancer screenings and the development of additional effective HPV vaccines, such as one for HPV 53 and 58. Additionally, gaining knowledge of HPV distribution may also inform us about ecological changes in HPV after the vaccination.