• Title/Summary/Keyword: M2 gene

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Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

Osteoblastogenic Activity of Tenebrio molitor Larvae Oil on the MG-63 Osteoblastic Cell (갈색거저리 유충 오일이 MG-63 조골세포 분화에 미치는 영향)

  • Seo, Minchul;Baek, Minhee;Lee, Joon Ha;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.29 no.9
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    • pp.1027-1033
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    • 2019
  • Recently, Korea has seen a rapid increase in the elderly population. As a result, osteoporosis, a geriatric disease, has become a social problem. To investigate the novel and natural materials for promoting osteoblastogenesis, we investigated the osteoblastogenic activity of Tenebrio molitor larvae oil (TMO) on the MG-63 preosteoblast cells. The cytotoxicity and proliferation effects of TMO on MG-63 cells were measured by MTS assay. There was no cytotoxicity up to $80{\mu}g/ml$. At 40 and $80{\mu}g/ml$ of TMO (treated for 48 hr), cell proliferation was elevated about 120% compared to the control. The osteoblastogenic activity of TMO was measured with alkaline phosphatase (ALP) activity at 5 days. Doses of 5 to $80{\mu}g/ml$ of TMO increased ALP activities significantly compared with the control. In addition, expression of ALP and Runx2 (osteoblastogenic markers) were markedly increased after treatment of TMO for 5 days. These results provide evidence that TMO promotes osteoblastogenesis by increasing the gene and protein expression of ALP and Runx2, and they suggest that TMO may be a potential agent for bone formation and preventing osteoporosis.

Studies on Genetic Analysis by the Diallel Crosses in $F_2$ Generation of Cowpea(Vigna sinensis savi.) (동부 Diallel Cross$ F_2$세대의 유전분석에 관한 연구)

  • Kim, J.H.;Ko, M.S.;Chang, K.Y.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.28 no.2
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    • pp.216-226
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    • 1983
  • Genetic studies on the $F_2$ generation of a set of half diallel crosses involving six cowpea varieties were conducted. by the randomized block design with three replications to determine combining ability, gene action and the relationships between parents and their $F_2$ hybrids. The 12 agronomic characters namely, days to flowering, days from flowering to maturity, days to maturity, diameter of stem, length of internode, number of branches per plant, length of pod, number of pods per plant, number of grains per pod, number of grains per plant, 100 grain weight and grain weight per plot were observed, and the $F_2$ generation of this diallel set of crosses was analysed for each character according to the method by Jinks and Hayman. The results obtained are summarized as follows: 1. Vr-Wr graphical analyses; The following seven characters, days to flowering, number of branches per plant, length of pod, number of pods per plant, number of grains per plant, 100 grain weight and grain weight per plot appeared to be partially dominant, and over dominance was found for days from flowering to maturity, days to maturity, length of internode and number of grains per pod. But diameter of stem indicated partial dominance near complete dominance. 2. Estimates of genetic variance components; In the degree of dominance,. eight characters, that is, days to flowering, days from flowering to maturity, days to maturity, length of internode, number of pods per plant, number of grains per pod, number of grains per plant and grain weight per plot were expressed larger than 1. And the characters, days from flowering to maturity, number of branches per plant and number of grains per plant as the degree of mean dominance ($H_1$/D) were found to be negative value over other characters. On the other hand, apprent asymmetry of dominance-recessive allele ($H_2$ /$4H_1$) produced comparatively estimates with lower value on days from flowering to maturity, length of internode, number of branches per plant and number of grains per pod. 3. Analyses of combining ability; Mean square value of GCA(general combining ability) appeared to be more important than those of SCA (specific combining ability) for most characters, and among them, grain weight per plot showed the highest mean square value in GCA and SCA. 4. Effect of combining ability; Variety 178 was expressed as the highest GCA effects in five characters (days to flowering days to maturity, number of pods per plant, number of grains per plant and grain weight per plot). SCA effects were differed from parents, characters and crosses, but crosses between TVu 1857 $\times$ TVu 2885 and TVu 2702 $\times$ J78 were shown to be highly with SCA effects on yield.

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Biogeochemical Remediation of Cr(VI)-Contaminated Groundwater using MMPH-0 (Enterobacter aerogenes) (MMPH-0 (Enterobacter aerogenes)에 의한 6가 크롬 오염 지하수의 생지화학적 정화)

  • Seo, Hyun-Hee;Rhee, Sung-Keun;Kim, Kang-Joo;Park, Eun-Gyu;Kim, Yeong-Kyoo;Chon, Chul-Min;Moon, Ji-Won;Roh, Yul
    • Economic and Environmental Geology
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    • v.45 no.2
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    • pp.105-119
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    • 2012
  • Indigenous bacteria isolated from contaminated sites play important roles to remediate contaminated groundwater. Chromium has the most stable oxidation states. Cr(VI) is toxic, carcinogenic, and mobile, but Cr(III) is less toxic and immobile. In this study, indigenous microorganism (MMPH-0) was enriched from Cr(VI) contaminated groundwater, and identified by 16S rRNA gene analysis. Using MMPH-0, the effect of stimulating with e-donors (glucose, lactate, acetate, and no e-donor control), respiration conditions, biomass, tolerance, and geochemical changes on Cr(VI) reduction were investigated in batch experiments for 4 weeks. The changes of Cr(VI) concentration and geochemical conditions were monitored using UV-vis-spectrophotometer and Eh-pH meter. And the morphological and chemical characteristics of MMPH-0 and precipitates in the effluents were characterized by TEM-EDS and SEM-EDS analyses. MMPH-0 (Enterobacter aerogenes) was able to tolerate up to 2000 mg/L Cr(VI) and reduce Cr(VI) under aerobic and anaerobic conditions. MMPH-0 performed faster and higher efficiency of Cr(VI) reduction with electron donors (over 70% after 1 week with e-donor, 10-20% after 4 weeks without e-donor). The changes of Eh-pH in effluents showing the tendency from oxidizing to reducing condition and a bit of acidic change in pH due to microbial oxidation of organic matters donating electrons and protons suggested the roles of MMPH-0 on Cr(VI) in the contaminated water catalyzing to transit geochemical stable zone for more stable $Cr(OH)_3$ or Cr(III) precipitates. TEM/SEM-EDS analyses of MMPH-0 and precipitates indicate direct and indirect Cr(VI) reduction: extracellular polymers capturing Cr component outside cells. These results suggested diverse indigenous bacteria and their biogeochemical reactions might enhance more effective and feasible remediation technology of redox sensitive heavy metals in metal-contaminated in groundwater.

THE ANTICANCER EFFECT OF PACLITAXEL($Taxol^{(R)}$) IN ORAL SQUAMOUS CELL CARCINOMA XENOGRAFT (이종 이식된 구강편평세포 암종에서 Paclitaxel ($Taxol^{(R)}$)의 항암 효과)

  • Kim, Ki-Hwan;Kim, Chul-Hwan;Han, Se-Jin;Lee, Jae-Hoon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.2
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    • pp.95-110
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    • 2006
  • The treatment for oral and maxillofacial carcinoma with chemotherapeutic agents is evaluated by many effective methods to reduce the tumor mass and cancer cell proliferation. However these chemotherapy have many serious side effects, such as bone marrow suppression, renal toxicity, G-I troubles. Therefore a possible approach to develop a clinically applicable chemotherapeutic agent is to screen anticancer activity of Taxol which is known to have very little side effect and have been used to breast cancer and ovarian carcinoma. Taxol is a new anti-microtubular anti-cancer agent extracted from the bark of the Pacific yew, Taxus brevifolia. Paclitaxel(Taxol) acts by promoting tubulin polymerization and over stabilizing microtubules agianst depolymerization. Despite the constant improvements of methods of the cancer treatment especially chemotherapy, the rate of cancer metastasis and recurrent are not decreased. Thus the investigation of new drug which have very little side effect and a possible clinically application continues to be a high priority. Considering that the Taxol have shown very effective chemotherapeutic agent with relatively low toxicity in many solid tumors, it deserves to evaluate its efficacy in oral squamous cell carcinoma. In this study, to investigate the in-vivo and in-vitro anti-cancer efficacy of Taxol in oral squamous cell carcinoma and lastly, the potency of Paclitaxel in the clinical application for oral cancer was evaluated. In vivo study, after HN22 cell line were xenografted in nude mice, the growth of tumor mass was observed, 3 mg/Kg taxol was injected intraperitoneally into nude mice containing tumor mass. The methods of these study were measurement of total volume of tumor mass, histopathologic study, immunohistochemical study, drug resistance assay, growth curve, MTT assay, flow cytometry, cDNA microarray in vivo and in vitro. The results were obtained as following. 1. The visual inspection of the experimental group showed that the volume of the tumor mass was slightly decreased but no significant difference with control group. 2. Ki-67 index was decreased at weeks 4 in experimental group. 3. Microscopic view of the xenografted tumor mass showed well differentiated squamous cell carcinoma and after Taxol injection, some necrotic tissue was seen weeks 4. 4. The growth curve of the tumor cells were decreased after 1day Taxol treatment. 5. According to the MTT assay, HN22 cell line showed relative drug resistancy above $5\;{\mu}g/ml$ concentrations of Taxol. 6. In drug resistance assay, the decrease of cell counts was seen relatively according to concentration. 7. In Flow cytometry, G2M phase cell arrests were seen in low concentration of the Taxol, while S phase cell arrests were seen in high concentration of the Taxol. 8. Using cDNA microarray technique, variable gene expression of ANGPTL4, TXNRD1, FAS, RRAGA, CTGF, CYCLINEA, P19, DUSP5, CEBPG, BTG1 were detacted in the oral squamous cell carcinoma cell after taxol treatment. In this study paclitaxel is effective against oral squamous cell carcinoma cell lines in vitro, but week effect was observed in vivo. So we need continuous study about anticancer effect of taxol in vivo in oral squamous cell carcinoma.

Correlation Between Primary Tuberculous Pleurisy and NRAMP1 Genetic Polymorphism (결핵성 흉막염 환자에서 NRAMP1 유전자 다형성에 대한 연구)

  • Kim, Je-Hyeong;Kim, Byung-Gyu;Jung, Ki-Hwan;Park, Sang-Myun;Lee, Sang-Youb;Lee, Sin-Hyung;Sin, Cheol;Cho, Jae-Youn;Shim, Jae-Jeong;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.155-165
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    • 2000
  • Background: The phagolysosomal function of alveolar macrophage against M. tuberculosis infection is influenced by Nramp1, which is encoded by the NRAMP1 gene. There are several genetic polymorphisms in NRAMP1, and these polymorphisms affect the innate host resistance through the defect in production and function of Nramp1. To investigate this relationship, the NRAMP1 genetic polymorphism in patients with primary tuberculous pleurisy was determined. Methods: Fifty-six primary tuberculous pleurisy patient, who were diagnosed by pleural biopsy, were designated to the pleurisy group and 45 healthy adults were designated to the healthy control group. Three genetic polymorphisms of NRAMP1, such as a single point mutation in intron 4(469+14G/C, INT4), a nonconservative single-base substitution at codon 543 that changes aspartic acid to asparagine(D543N) and a TGTG deletion in the 3' untranslated region(1729+55delI4, 3'UTR), were determined. Polymerase chain reaction(PCR) and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) were used. Results: The frequencies of mutant genotypes of INT4 and 3'UTR were significantly high in pleurisy group(p=0.001, p=0.023). But the frequencies of D543N were not significantly different between the two groups(p=0.079). The odds ratios, which are a comparison with wild genotype for determining mutant genotypes, were 8. 022(95% confidence interval=2.422-26.572) for INT4 and 5.733(95% confidence interval = 1.137~28.916) for 3'UTR ; these were statistically significant But the ratio for D543N was not significant In the combined analysis of the INT4 and 3'UTR polymorphisms, the odds ratios were 6.000(95% confidence interval = 1.461~24.640) for GC/++ genotype and 14.000(95% confidence interval=1.610~121.754) for GC/+del when compared with GG/++ homozygotes ; these were statistically significant. Conclusion: Among the NRAMP1 genetic polymorphisms, a single point mutation in intron 4(469+14G/C, INT4) and a TGTG deletion in the 3' untranslated region(1729+55del4, 3'UTR) were closely related to the primary tuberculous pleurisy.

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Characteristics of Wheat Flour Dough and Noodles with Amylopectin Content and Hydrocolloids (아밀로펙틴 함량 변화와 하이드로콜로이드 첨가에 의한 밀가루 반죽 및 국수의 특성)

  • Cho, Young-Hwa;Shim, Jae-Yong;Lee, Hyeon-Gyu
    • Korean Journal of Food Science and Technology
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    • v.39 no.2
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    • pp.138-145
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    • 2007
  • The effects of amylopectin and hydrocolloid (locust bean gum and guar gum) content on wheat flour dough and noodle properties were investigated. As the amount of amylopectin increased, the water absorption rate (farinograph), the tension (tension test), the gel stability (freeze-thawing treatment), and the springiness and the cohesiveness (TPA) increased, but the pasting temperature (RVA), the lightness and yellowness (color measurement), and the hardness (TPA) tended to decrease. In sensory evaluations, the scores for cohesiveness, springiness, and acceptability of cooked noodle increased as the proportion of amylopectin increased. The proper combination of amylose/amylopectin ratio and hydrocolloids improved the freeze-thaw stability and the sensory acceptability of wheat flour dough and noodle.

Molecular Diagnosis of Streptococcus pneumoniae in Middle Ear Fluids from Children with Otitis Media with Effusion (삼출성 중이염 소아의 중이액에서 폐구균의 분자적 진단)

  • Byun, Sung Wan;Kim, Han Wool;Yoon, Seo Hee;Park, In Ho;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.106-112
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    • 2015
  • Purpose: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. Methods: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. Results: The detection limit of the PCR assay was approximately $10^4$ colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over $10^6$ CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). Conclusions: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA -specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.

Studies on the Generation-to-Generation Transmission of Cytoplasmic Polyhedrosis Virus and the Effect of Their Activation on the Induction in the Next Generation in the Silkworm, Bombyx mori L. (Virus의 경란전염이 차대의 잠작에 미치는 영향에 관한 연구)

  • 임종성;김근영
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.85-92
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    • 1974
  • Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2% or 3%). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,

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Study on the Clonality of Endothelial Cell Proliferation in Plexiform Lesions in Patients with Pulmonary Hypertension Associated with CREST Syndrome (CREST 증후군에 동반된 폐고혈압 환자에서 총상병변내 내피세포 증식의 클론성에 관한 연구)

  • Lee, Sang-Do;Jeon, Yong-Gam;Lee, Ji-Hyun;Shim, Tae-Sun;Lim, Chae-Man;Koh, Yun-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Tuder, Rubin M.
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.2
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    • pp.150-160
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    • 1999
  • Background: The CREST syndrome is an indolent form of progressive systemic sclerosis. Although its clinical progress is indolent, pulmonary hypertension(PH) associated with CREST syndrome have grave prognosis with over 40 percent mortality rate at 2 year follow-up. But the pathogenesis of pulmonary hypertension in this disease is not known, and classified as either primary or secondary PH. Clonality of endothelial cell proliferation in plexiform lesion is a molecular marker which allows distinction between primary and secondary PH. We performed this study to know whether the PH associated with CREST syndrome is a variant of primary PH or is a secondary PH. Methods: We assessed the X-chromosome inactivation based on the methylation pattern of the human androgen-receptor gene by PCR(HUMARA). Endothelial cells in plexiform lesions from female patients(n=3) with PH associated with CREST syndrome were microdissected from paraffin blocks. Vascular smooth muscle cells and lung parenchyma were also microdissected for clonality studies. Results: The proliferating endothelial cells in 14 plexiform lesions were all polyclonal. Similarly proliferated smooth muscle cells from 5 vessels with medial hypertrophy were also polyclonal. Conclusion: These results suggest that the pulmonary hypertension associated with CREST syndrome has different pathogenesis from primary PH and to be classified as secondary PH.

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