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The Significance of Ku70 Expression in Patients with Nasopharyngeal Carcinoma (비인강암에서 방사선치료의 예측인자로써 Ku70 발현의 의의)

  • Song Si Yeol;Lee Sang-wook;Yu Eunsil;Cho Kyung-Ja;Park Jin-hong;Kim Sang Yoon;Nam Soon Yuhl;Lee Bong-Jae;Kim Sung Bae;Choi Seung-Ho;Ahn Seung Do;Shin Seong Soo;Choi Eun Kyung
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.1-8
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    • 2005
  • Purpose : The objective of this study was to determine whether the expressions of the two components of DNA-dependent protein kinase, Ku70 and DNA-PKcs, influence the response to radiotherapy (RT) and outcome of treatment of non-disseminated nasopharyngeal carcinoma (NPC) in patients who received definitive RT. Materials and Methods : Sixty-six patients with NPC who were treated with radiotherapy alone or with concurrent chemotherapy between June 1995 and December 2001 were divided into groups based on the levels of immunoreactivity for Ku70 and DNA-PKcs in pretreatment biopsy specimens. The over-expression of Ku70 or DNA-PKcs groups Included patients whose biopsy specimens showed at least 50% immunopositive tumor cells; patients in which less than 50% of the tumor cells in the biopsy tissues were immunopositive were placed in the low Ku70 and DNA-PKcs groups. The immunoreactivities for Ku70 and DNA-PKcs were retrospectively compared with the sensitivity of the tumor to radiation and the patterns of therapy failure. Univariate analyses were peformed to determine the prognostic factors that influenced locoregional control of NPC. Results : The five-year locoregional control rate was significantly higher in the low Ku70 group (Ku(-)) (85%) than in the high Ku70 group (Ku(+)) (42%) (p=0.0042). However, there were no differences in the metastases-free survival rates between the two groups (Ku70 (+), 82%; Ku70 (-), 78%; p=0.8672). Univariate analysis indicated that the over-expression of Ku70 surpassed other well-known predictive clinocopathologic parameters as an Independent prognostic factor for locoregionai control. Eighteen of 22 patients who had locoregional recurrences of the tumor displayed an over-expression of Ku70. No significant association was found between the level of DNA-PKcs expression and the clinical outcome. Conclusion : Our data suggest that the level of Ku70 expression can be used as a molecular marker to predict the response to RT and the locoregional control after RT and concurrent chemotherapy in patients with non-disseminated NPC.

Correlation of serum total hydroperoxide levels and diseases of prematurity (미숙아 합병증 발생과 혈중 활성 산화물 농도와의 관계)

  • Kim, Shin-Hye;Hur, Hae Young;Lee, Kyu Hyoung;Moon, Ja Young;Chae, Kyu Young
    • Clinical and Experimental Pediatrics
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    • v.50 no.8
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    • pp.746-752
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    • 2007
  • Purpose : Total hydroperoxide (TH), free radical-mediated oxidation product can be used as a measure of free radical injury. The aim of the present study was to see if preterm newborns are at increased risk for oxidative stress compared with term newborns, and to determine whether oxidative stress during postnatal first 1 week is associated with clinical outcomes in preterm infants. Methods : Serum TH levels of preterm infants (n=39) were compared with those of term infants (n=24) on the postnatal day 1. Among the preterm infants, serum TH levels of uncomplicated group (n=23) were also compared with those of complicated group (n=16) who developed oxygen radical related diseases on the postnatal day 1 and 7. Retrospective analysis was performed to find out risk factors for oxygen radical injuries based on birth history, laboratory data, neuroimaging findings and clinical progress in two preterm groups. Results : Serum TH levels on postnatal day 1 were higher in the preterm infant group than the term infant group. Serum TH levels on postnatal day 1 in the complicated preterm infant group were significantly higher compared with uncomplicated group, but there was no significant difference in serum TH levels on postnatal day 7. Also, there was no significant difference in serum TH levels between uncomplicated preterm infants and term infants. Serum TH level on postnatal day 1 was independently associated with higher morbidity after adjusting for gestational age, Apgar score (5 min), arterial blood gas analysis. Conclusion : Complicated preterm newborns are at increased risk for oxidative stress compared with uncomplicated newborns and term newborns. Oxidative injury during the prenatal or postnatal day 1 is associated with adverse outcomes in preterm infants. Elevated TH levels on postnatal day 1 may have a value to predict clinical outcomes in preterm infants.

Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11 (고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사)

  • Woo, Sang-Min;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.50 no.2
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    • pp.95-100
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    • 2007
  • The cellulase gene of Bacillus licheniformis K11 which has plant growth-promoting activity by auxin and antagonistic ability by siderophore was cloned in pUC18 using PCR employing heterologous primers. The 1.6kb PCR fragment contained the full sequence of the cellulase gene, denoted celW which has been reported to encode a 499 amino acid protein. Similarity search in protein data base revealed that the cellulase from B. licheniformis K11 was more than 97% identical in amino acid sequence to those of various Bacillus spp. The cellulase protein from B. licheniformis K11, overproduced in E. coli DH5${\alpha}$ by the lac promoter on the vector, had apparent molecular weight of 55 kDa upon CMC-SDS-PAGE analysis. The protein not only had enzymatic activity toward carboxymethyl-cellulose (CMC), but also was able to degrade insoluble cellulose, such as Avicel and filter paper (Whatman$^{\circledR}$ No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. Consequently B. licheniformis K11 was able to suppress the peperblight causing P. capsici by its cellulase. Biochemical analysis showed that the enzyme had a maximum activity at 60$^{\circ}C$ and pH 6.0. Also, the enzyme activity was activated by Co$^{2+}$ of Mn$^{2+}$ but inhibited by Fe$^{3+}$ or Hg$^{2+}$. Moreover, enzyme activity was not inhibited by SDS or sodium azide.

Effect of Gastrodiae elata Blume Components on Systolic Blood Pressure and Serum Lipid Concentrations in Spontaneously Hypertensive Rats Fed High Fat Diet (천마성분이 본태성고혈압쥐의 혈압과 혈청지질 농도에 미치는 영향)

  • Hong, Hee-Do;Shim, Eun-Jung;Kim, Kyung-Im;Choi, Sang-Yoon;Han, Chan-Kyu
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.2
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    • pp.174-179
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    • 2007
  • This study was carried out to investigate the effect of Gastrodiae elata Blume fractions on systolic blood pressure and serum lipid profiles in spontaneously hypertensive rats (SHR/NCrj) fed high fat diet. Twenty-four males SHR weighing approximately 160 g were randomly divided into four groups; A (low molecule, GR-1), B (polysaccharide, GR-2), C (protein, GR-3) fractions of G. elata Blume, respectively, and D (high fat diet as control). After orally tube feeding the fractions of G. elata Blume, there were no differences in final body weights among the treatment groups. Diet intake was somewhat high in the control group (D), but there were no significant differences in feed efficiency ratios. In terms of serum lipid profiles, total-cholesterol level was statistically higher in the control group (D) than in G. elata Blume fraction groups (p<0.05). Triglyceride levels of low molecule (A) and polysaccharide (B) groups were lower by 16% and 11%, respectively than that of the control group (D). HDL-cholesterol level was remarkably higher (p<0.05), whereas LDL-cholesterol level was significantly lower (by 25%) in the group B as compared to the control group (D). Atherogenic index (AI) of G. elata Blume fraction groups were significantly lower than in the control group (p<0.05). Reference blood pressure (RBP) showed an average of $180\sim190mmHg$ at 8 weeks old after 3 weeks on feeding high fat diet. Compared with RBP, final blood pressure of treatment groups (35 days after feeding the fractions of G. elata Blume gractions) were decreased by 1.7% (A), 5.5% (B) and 3.6% (C), respectively, but the control group (D) contrarily showed an increase of 2.6%. Especially, final systolic blood pressure of the polysaccharide group (B) was lower by 22 mmHg than that of the control group (D). From these findings, it can be suggested that polysaccharide fraction may improve blood serum lipids and should be considered as effective in lowering of blood pressure.

Sulfur Dioxide, Mineral Contents and Physicochemical Properties Generated during Manufacture of Bamboo Salt (죽염 제조공정에 따른 이산화황, 미네랄 함량 및 이화학적 특성)

  • Kim, Hag-Lyeol;Lee, Seong-Jae;Lee, Jung-Hee;Kim, In-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.8
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    • pp.1248-1256
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    • 2014
  • The purpose of this study was to investigate the mechanisms of behind $SO_2$ formation and elevated cause of reducing power in purple bamboo salt (PBS) along with an analysis of physicochemical properties, content of sulfur compounds, oxidation reduction potential (ORP), mineral contents of salt type (MSS, mudflat solar salt; BS, bamboo salt), and addition of raw bamboo (RB). $SO_2$ content of 630 ppm was detected in PBS. $SO_2$ was not detected in MSS, BS, or RB, whereas $SO_2$ (782 ppm) from $K_2SO_4$ was detected after heating a NaCl, KCl, $MgCl_2$, $MgSO_4$, MgO, $CaCl_2$, $K_2SO_4$, and $FeSO_4$ with RB. $SO_2$ content of BS increased with baking time, and it originated from BSRB1 (13.88 ppm) to BSRB4 (109.13 ppm). $SO_3{^{2-}}$ originated only from MSSRB4 and BSRB2~BSRB4. Sulfate ion content decreased along with increasing $SO_2$ and sulfite ion contents. ORP increased with baking time of MSS and BS, and it was present at higher levels in BSRB4 (-211.40 mV) of BS than MSS. Insoluble content was higher in BS than MSS. Further, Ca, K, and Mg ion contents decreased in MSS and increased in BS with baking time. BSRB4 had 1.4 fold higher levels of Ca, 1.5 fold higher levels of Mg, and 1.8 fold higher levels of K than BS. Li, Al, Mn, Fe, and Sr in MSS as well as Al, Fe, and Ni in BS increased with baking time. Anions (Cl, $NO_3$, and Br) and heavy metals (Pb, Cd, Hg, and As) between MSS and BS were not significantly different. These results suggest that the reducing power of BS was due to $SO_2$ and sulfite ion. To increase the amounts of these compounds and reducing power, higher melting temperature and longer baking time are necessary along with BS, which is created by the addition of RB to roasted salt.

Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

Expressions of Matrix Metalloproteinase-9 and Tissue Inhibitor of Metalloproteinase-2 with Changes of Interleukin-6 and Interleukin-18 in Atherosclerotic Lesions of Hypercholesterolemic Rabbits (고콜레스테롤혈증 가토의 죽상경화성 병변에서 Interleukin-6와 Interleukin-18의 변화 및 Matrix Metalloproteinase-9과 Tissue Inhibitor of Metalloproteinase-2의 발현)

  • 권영무;김성숙;장봉현
    • Journal of Chest Surgery
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    • v.35 no.6
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    • pp.407-419
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    • 2002
  • Atherosclerosis is a chronic inflammatory disease of the arterial wall characterized by progressive accumulation of lipids, cells, and extracellular matrix. Matrix metalloproteinases(MMPs) and tissue inhibitor of metalloproteinases(TIMPS) contribute to vascular matrix remodeling in atherosclerosis, and some cytokines may play role in the synthesis or activation of MMPs or TIMPs. Material and Method: We produced experimental atherosclerotic plaques in 9 rabbits by atherogenic hypercholesterol diet for 12 weeks, and 10 other rabbits were used as control group with standard laboratory chow, At that time, 19 rabbits were sacrificed and aorta, coronary arteries and blood specimens were prepared. The expressions of MMP-9, TIMP-2 and interleukin(IL)-18, and the bioactivity of IL-6 were investigated with H&E stain, immunohistochemical stain, immunoblotting(Western blot analysis), and bioassay. Result: Serum cholesterol in the experimental group increased up to 1258$\pm$262 mg/dL(control group: 41$\pm$7 mg/dL). All experimental group showed well-developed atherosclerotic plaques in aorta and coronary artery. The expression of MMP-9 in aorta and coronary artery of the experimental group showed significant increase than that of the control group by immunohistochemistry. Among the experimental group, complicated lesions with intimal rupture or complete luminal occlusion, demonstrated stronger expression of MMP-9. Interestingly, there was no difference in expression of TIMP-2 between the experimental and the control group. These findings were confirmed by Western blot analysis. The bioassay revealed significant up-regulation of serum bioactivity of IL-6 in the experimental group(4819.60$\pm$2021.25 IU/$m\ell$) compared to that of IL-6 in the control group(27.20 $\pm$ 12.19 IU/$m\ell$). IL-18 was expressed in all atherosclerotic plaques, whereas little or no expression was detected in the control group. Conclusion: The increased MMP-9 expression along with the unchanged TIMP-2 expression seem to be contributory factors in extracellular matrix degradation in atherosclerosis. Focal overexpression of MMP-9 may promote plaque destabilization and cause complications of atherosclerotic plaques such as thrombosis with/without acute coronary syndrome. Elevation of IL-6 and IL-18 may be more than just markers of atherosclerosis but actual participants in lesion development. Identification of critical regulatory pathway is important to improve the understanding of the cellular and molecular basis of atherosclerosis and may open the way for novel therapeutic strategies.

Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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Germinated Rhynchosia nulubilis Hydrolysate Ameliorates Dexamethasone-induced Muscle Atrophy by Downregulating MAFbx Expression in C2C12 Cells and C57BL/6 Mice (발아 서목태 가수분해물의 근위축 억제 효과)

  • Won Keong Lee;Eun Ji Kim;Sang Gon Kim;Young Min Goo;Young Sook Kil;Seung Mi Sin;Min Ju Ahn;Min Cheol Kang;Young-Sool Hah
    • Journal of Life Science
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    • v.33 no.3
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    • pp.277-286
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    • 2023
  • Sarcopenia is the age-related loss of muscle mass and function. It is a natural part of aging and can lead to decreased mobility and increased frailty. The ubiquitin-proteasome pathway, which is involved in muscle protein degradation, is closely linked to sarcopenia. Germinated Rhynchosia nulubilis hydrolysate (GRH) has been reported to have anti-inflammatory and antioxidant properties, but there have been no reports on its inhibitory effect on muscle reduction. However, no study has yet explored the relationship between GRH and muscle loss inhibition. In this study, we evaluated the effects of GRH on muscle atrophy inhibitory activity in dexamethasone (Dexa)-induced muscle atrophy C2C12 myotubes and mouse models. Moreover, we identified a molecular pathway underlying the effects of GRH on skeletal muscle. May Grunwald-Giemsa staining showed that the length and area of myotubes increased in the groups treated with GRH. In addition, the GRH-treated group significantly reduced the expression of muscle ring finger protein 1 and muscular atrophy F-box (MAFbx) in the Dexa-induced muscular atrophy C2C12 model. GRH also improved muscle strength in C57BL/6 mice with Dexa-induced muscle atrophy, resulting in prolonged running exhaustive time and increased grip strength. We found that muscle strengthening by GRH was correlated with a decreased expression of the MAFbx gene in mouse muscle tissue. In conclusion, GRH can attenuate Dexa-induced muscle atrophy by inhibiting the ubiquitin-proteasome pathway via downregulation of the MAFbx gene expression.

A prediction study on the number of emergency patients with ASTHMA according to the concentration of air pollutants (대기오염물질 농도에 따른 천식 응급환자 수 예측 연구)

  • Han Joo Lee;Min Kyu Jee;Cheong Won Kim
    • Journal of Service Research and Studies
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    • v.13 no.1
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    • pp.63-75
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    • 2023
  • Due to the development of industry, interest in air pollutants has increased. Air pollutants have affected various fields such as environmental pollution and global warming. Among them, environmental diseases are one of the fields affected by air pollutants. Air pollutants can affect the human body's skin or respiratory tract due to their small molecular size. As a result, various studies on air pollutants and environmental diseases have been conducted. Asthma, part of an environmental disease, can be life-threatening if symptoms worsen and cause asthma attacks, and in the case of adult asthma, it is difficult to cure once it occurs. Factors that worsen asthma include particulate matter and air pollution. Asthma is an increasing prevalence worldwide. In this paper, we study how air pollutants correlate with the number of emergency room admissions in asthma patients and predict the number of future asthma emergency patients using highly correlated air pollutants. Air pollutants used concentrations of five pollutants: sulfur dioxide(SO2), carbon monoxide(CO), ozone(O3), nitrogen dioxide(NO2), and fine dust(PM10), and environmental diseases used data on the number of hospitalizations of asthma patients in the emergency room. Data on the number of emergency patients of air pollutants and asthma were used for a total of 5 years from January 1, 2013 to December 31, 2017. The model made predictions using two models, Informer and LTSF-Linear, and performance indicators of MAE, MAPE, and RMSE were used to measure the performance of the model. The results were compared by making predictions for both cases including and not including the number of emergency patients. This paper presents air pollutants that improve the model's performance in predicting the number of asthma emergency patients using Informer and LTSF-Linear models.