톡소플라스마 곤디의 세포내 배양에 있어서 세포 주에 따른 감수성 비교

박병규(Byung-Kiu Park),문형로(Hyung Ro Moon),유재란(Jae-Ran Yu),국진아(Jina Kook),채종일(Jong-Yil Chai),이순형(Soon-Hyung Lee) 대한기생충학열대의학회 1993 The Korean Journal of Parasitology Vol.31 No.3

유방암세포주에서 고농도 5-fluorouracil의 세포주기 조절효과

장정순,양중일,장세호,이원섭,이종석,안명주,박병규,Jang, Joung Soon,Yang, Jung Ill,Chang, Seho,Lee, Won Sup,Lee, Jong Seok,Ahn, Myung-Ju,Park, Byung-Kiu 대한면역학회 2002 Immune Network Vol.2 No.1

Background: Chemotherapy with 5-fluorouracil (5-FU) has been one of the mainstay in breast cancer treatment. The effects of high dose 5-FU on cell cycle regulation were studied in breast caner cells. Methods: A breast cancer cell line MCF-7 was used. Protein expressions of G1/S cyclins, $p21^{Waf1/Cip1}$, cdk2, E2F1 and retinoblastoma were tested by western blot analysis. Immunoprecipitation and immune complex kinase assay were done for the assessment of E2F1/RB interacton and the activity of cdk2 respectively. Results: $p21^{Waf1/Cip1}$ expression was barely detectable in control cells. With addition of 5-FU level of $p21^{Waf1/Cip1}$ were induced and cyclin D3 level was decreased as cell growth decreases. In accordance with increased expression of $p21^{Waf1/Cip1}$, cyclin E-associated cdk2 kinase activity was reduced. Retinoblastoma protein (RB) became dephosphorylated and E2F-1 binding activity with RB was increased. Conclusion: In this situation of high concentration of 5-FU breast cancer cells tend to be G1/S cell cycle arrested. Overexpression of $p21^{Waf1/Cip1}$ and dephosphorylation of RB may mediate the effectss of 5-FU by inhibiting E2F-1 activity, which contributes to G1/S cell cycle arrest. These results could be an indicating landmark for further study of high dose chemotherapy with 5-FU.

암 환자의 혈장 Transforming Growth Factor-β1 농도

전지현(Ji-Hyun Jeon),이시은(Si-Eun Lee),이수진(Sue-Jin Lee),박찬후(Chan-Hoo Park),장정순(Joung Soon Jang),하우송(Woo Song Ha),박순태(Soon Tae Park),박병규(Byung-Kiu Park) 대한의생명과학회 1999 Biomedical Science Letters Vol.5 No.2

한국인의 대표적인 성인 고형 종양인 위암, 간암, 유방암과 소아 백혈병 및 2종의 소아 고형 종양 환자로부터 혈장 transforming growth factor-β1 (TGF-β1) 농도를 sandwich ELISA 분석법을 이용해 측정함으로써 TGF-β1을 이 질환들에 대한 새로운 종양표지자 (tumor marker)로 사용할 수 있는지 검토하였다. 또한 연령 및 성별에 따른 혈장 TGF-β1 농도의 정상치를 조사하였다. 신생아에서 70대까지 혈장 TGF-β1 농도의 차이는 없었고 남녀간의 차이도 없었다. 위암 환자의 혈장 TGF-β1 농도는 16.0±6.8 ng/㎖ (평균±표준편차)로 정상 대조군의 TGF-β1 농도 (8.3±5.0 ng/㎖) 보다 유의하게 높았으나 간암, 유방암 환자의 혈장 TGF-β1 농도는 대조군과 차이가 없었다. 그리고 위암 환자 16명, 간암 환자 8명, 유방암 환자 7명 중 각각 7명 (43.7%), 1명 (12.5%), 1명 (14.3%)에서만 혈장 TGF-β1 농도가 증가되었다. 5명의 소아 백혈병 환자에서는 관해 (remission) 여부와 상관없이 혈장 TGF-β1 농도가 모두 정상 범위에 있었으나 2명의 소아 고형암 환자에서는 종양 절제 전에는 혈장 TGF-β1 농도가 높았다가 절제 후 정상으로 떨어졌다. 결론적으로 1) 정상인의 혈장 TGF-β1 농도는 연령 및 성별에 따른 차이가 없다는 것을 알 수 있었고, 2) 성인 고형암인 위암, 간암, 유방암에서는 낮은 민감도로 인해 TGF-β1을 진단을 위한 선별 검사로 이용하기에는 부적절한 것으로 판단되었으며, 3) 정상 대조군보다 혈장 TGF-β1 농도가 높았던 위암 환자와 종양 절제 전후로 혈장 TGF-β1 농도가 민감하게 변했던 소아 고형암 환자에 대해서는 향후 표본 수를 늘려 부가적인 연구를 해야 할 것으로 사료된다. To evaluate the usefulness of transforming growth factor-β1 (TGF-β1) as a new tumor marker, we determined the plasma TGF-β1 levels using sandwich ELISA assay in cancer patients. Patients with three most common adult cancers in Korea (stomach, liver and breast cancer) and children's cancers (leukemia and two kinds of solid tumor) were enrolled for the study. Furthermore, 39 individuals were subjected to age and sex-stratified plasma TGF-β1 analysis. No statistical difference was demonstrated with respect to age or sex. The mean plasma TGF-β1 level (16.0 ng/㎖) of stomach cancer patients was significantly higher than that (8.3 ng/㎖) of controls. However, there was no difference among the mean plasma TGF-β1 levels of liver, breast cancer patients and controls. Seven of 16 patients (43.7%) with stomach cancer, one of 8 (12.5%) with liver cancer, and one of 7 (14.3%) with breast cancer showed higher TGF-β1 levels compared to controls. Plasma TGF-β1 concentrations of five leukemic children remained in the normal range regardless of the remission state. In contrast, initial high TGF-β1 levels from two children with solid tumors returned to normal range on surgical resection of tumors. From the above results, we could conclude that plasma TGF-β1 levels of apparently healthy individuals seem to be rather constant irrespective of difference in age or sex, and the plasma TGF-β1 has the limited value as a screening test for the diagnosis of aforementioned adult cancers because of its low sensitivity. Finally, additional studies need to be pursed for the large number of stomach cancer and pediatric solid tumor patients in order to reach a secure conclusion on the usefulness of plasma TGF-β1 as a tumor marker in these patients.

암 환자의 혈장 Transforming Growth Factor-β1 농도

박병규,하우송,이시은,이수진,박순태,박찬후,전지현,장정순 THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCINE 1999 Journal of biomedical laboratory sciences Vol.5 No.2

한국인의 대표적인 성인 고형 종양인 위암, 간암, 유방암과 소아 백혈병 및 2종의 소아 고형 종양 환자로부터 혈장 transforming growth factor-ß1 (TGF-ß1) 농도를 sandwich ELISA 분석법을 이용해 측정함으로써 TGF-ß1을 이 질환들에 대한 새로운 종양표지자 (tumor marker)로 사용할 수 있는지 검토하였다. 또한 연령 및 성별에 따른 혈장 TGF-ß1 농도의 정상치를 조사하였다. 신생아에서 70대까지 혈장 TGF-ß1 농도의 차이는 없었고 남녀간의 차이도 없었다. 위암 환자의 혈장TCF-ß1 농도는 16.0±6.8 ng/ml (평균 ±표준편차)로 정상 대조군의 TGF-ß1 농도 (8.3 ±5.0 ng/ml) 보다 유의하게 높았으나 간암, 유방암 환자의 혈장 TGF-ß1 농도는 대조군과 차이가 없었다. 그리고 위암 환자 16명, 간암 환자 8명, 유방암 환자 7명 중 각각 7명 (43.7%), 1명 (12.5%), 1명 (14.3%)에서만 혈장 TGF-ß1 농도가 증가되었다. 5명의 소아 백혈병 환자에서는 관해 (remission) 여부와 상관없이 혈장 TGF-ßl 농도가 모두 정상 범위에 있었으나 2명의 소아 고형암 환자에서는 종양 절제 전에는 혈장TGF-ß1 농도가 높았다가 절제 후 정상으로 떨어졌다. 결론적으로 1)정상인의 혈장 TGF-ßl 농도는 연령 및 성별에 따른 차이가 없다는 것을 알 수 있었고, 2)성인 고형암인 위암, 간암, 유방암에서는 낮은 민감도로 인해 TGF-ß1을 진단을 위한 선별 검사로 이용하기에는 부적절한 것으로 판단되었으며, 3) 정상 대조군보다 혈장 TGF-ß1 농도가 높았던 위암 환자와 종양 절제 전후로 혈장 TGF-ß1 농도가 민감하게 변했던 소아 고형 암 환자에 대해서는 향후 표본 수를 늘려 부가적인 연구를 해 야 할 것으로 사료된다. To evaluate the usefulness of transforming growth factor-ß1 (TGF-ß1)as a new tumor marker, we determined the plasma TGF-ß1 levels using sandwich ELISA assay in cancer patients. Patients with three most common adult cancers in Korea (stomach, liver and breast cancer) and children's cancers (leukemia and two kinds of solid tumor) were enrolled for the study. Furthermore, 39 individuals were subjected to age and sex-stratified plasma TGF-ß1 analysis. No statistical difference was demonstrated with respect to age or sex. The mean plasma TGF-ß1 level (16.0 ng/ml) of stomach cancer patients was significantly higher than that (8.3ng/ml) of controls. However, there was no difference among the mean plasma TGF-ß1 levels of liver, breast cancer patients and controls. Seven of 16 patients (43.7%) with stomach cancer, one of 8 (12.5%) with liver cancer, and one of 7 (14.3%) with breast cancer showed higher TGF-ß1 levels compared to controls. Plasma TGF-ß1 concentrations of five leukemic children remained in the normal range regardless of the remission state. In contrast, initial high TGF-ß1 levels from two children with solid tumors returned to normal range on surgical resection of tumors. From the above results, we could conclude that plasma TGF-ß1 levels of apparently healthy individuals seem to be rather constant irrespective of difference in age or sex, and the plasma TGF-ß1 has the limited value as a screening test for the diagnosis of aforementioned adult cancers because of its low sensitivity. Finally, additional studies need to be pursed for the large number of stomach cancer and pediatric solid tumor patients in order to reach a secure conclusion on the usefulness of plasma TGF-ß1 as a tumor marker in these patients.

선천성 부신 과형성을 초래하는 Steroid 21-hydroxylase 유전자 ( CYP21A2 )의 분자 유전학적 연구 (1)

박병규,우향옥,유한욱 대한내분비학회 1994 Endocrinology and metabolism Vol.9 No.3

Congenital adrenal hyperplasia, especially due to steroid-12-hydroxylase(P450c21) deficiency, is one of the most common autosomal recessive inborn errors at adrenal steroidogenesis in Korean. Molecular genetic analysis has demonstrated that there are two steroid 21-hydroxylase genes, CYP21A1P and CYP21A2. The CYP21A2 gene encodes P450c21, whereas the CYP21A1P gene is a pseudogene. Since there is 98 percent homology between the CYP21A1P and CYP21A2 gene in nucleotide sequences, it has hampered the characterization of molecular defects in the CYP21A2 gene. In this study, efforts have been made to selectively PCR amplify the CYP21A2 gene and test feasibility of DNA microextraction from Guthrie card for prospective use of molecular screening. This study was also aimed at investigating deletion mutations in P450c21 deficient patients, as well as allele frequencies and average heterozygosity of exon 1 A/C polymorphism in Korean newborns. Genomic DNAs were obtained from Guthrie cards of 50 Korean newborns by microextraction method and these DNAs were analyzed by PCR-allele specific oligonucleotide(ASO) hybridization. First part of the CYP21A2 gene has been successfully amplified and digested by restriction enzyme using Taq I or Kpn I, subsequently run on 1.5% agarose gel to confirm its specificity. The anterior 1141 bp PCR product was utilized to examine the frequency and average heterozygosity of exon 1 A/C polymorphism in 100 alleles by ASO dot blot hybridization. Amplified genomic DNAs from four P450c21 deficient patients out of three families were screened by PCR to see if any one has complete deletion of the CYP21A2 gene. The results were as follows; 1) The average 1230ng of genomic DNA was obtained form single semi-circled Guthrie card of 1/2 inch diameter by microextraction method, which has been successfully used for DNA analysis. 2) The PCR amplified anterior 1141 bp product from the CYP21A2 gene was digested by Kpn I, generating 309 bp, 832 bp fragments, not by Taq I, indicating its specificity. 3) The frequencies of exon 1 nucleotide 138 A/C polymorphism in Korean population were 0.81, 0.91 respectively, and average heterozygosity was 0.31. 4) None of four P450c21 deficient patients turned out to carry complete deletion of the CYP21A2 gene based on selective PCR amplification of the CYP21A2 gene. In conclusion, dried blood spots from Guthrie card can be sued for DNA analysis because of easy sample collection, bandling, shipment, and DNA extraction feasibility. The selective PCR amplification of the CYP 21A2 gene will pave the way for molecular characterization in P450c21 deficient patients. The exon 1 A/C polymorphism can by efficiently used for molecular diagnosis of P450c21 deficiency in informative families, though it has a drawback of handling radioactive material(J Kor Soc Endocrinol 9: 219-227, 1994).

2개월 남아에서 발병한 Malignant Ectomesenchymoma

황지영,서지현,김점수,김용숙,이수진,박찬후,김유경,박선후,이정희,안인옥,정기현,박병규 대한소아과학회 2001 小兒科 Vol.44 No.8

요중 성장 호르몬치와 인슐린 , L - dopa 를 이용한 성장 호르몬 유발검사에서의 혈중 성장 호르몬 최고치와의 상관관계

김철호,박병규,정우영 대한내분비학회 1995 Endocrinology and metabolism Vol.10 No.4

To investigate the correlation between urinary growth hormone(GH) level and peak serum GH level, urinary GH value measured by overnight collection of urine for 10 hours and serum GH value in response to GH provocation test using insulin and L-dopa were measured in 9 cases of GH complete deficiency(GCD), 19 cases of GH partial deficiency(GPD) and 40 cases of GH normal short stature(GHN). Urinary GH values were measured by the EIA method using PICOIA HGH plate(Joo Woo Pharmaceutical Co., Japan). Urinary GH was expressed in terms of nanograms per gm creatinine(ng/gCr). Serum GH was measured by immunoradiometric assay using "Daiichi kit"(Je Il Pharmaceutical Co., Japan). Wilcoxon ranked sum test and student's t-test were used to assess the significance of differences between the groups of the patients. The correlation between urinary GH level and peak serum GH level was assessed by the parametric Pearson correlation test. The correlation between peak serum GH level in GH provocation test using insulin and urinary GH level measured by overnight 10 hours collection method showed statistically significant results in all the patients(Y=0.464072X +9.208044, r=0.48987, p=0.0001) and in the GH deficiency groups(GCD+GPD) (Y=0.924659X +9.2385509, r=0.80437, p=0.0001). In case of L-dopa stimulation test, urinary GH values were also positively correlated with peak serum GH level when all the patients were participated(Y=0.572988X +8.312993, r=0.58212, p=0.0001). In contrast, no correlation was found when patients were confined to GH deficiency group(GCD+GPD)(Y=0.127712X +8.3129939, r=0.08044, p=0.6841)(J Kor Soc Endocrinol 10: 370-376, 1995).

성장호르몬 결핍증 환아에서 성장호르몬 분비안자에 대한 성장호르몬 분비 양상

문형로,양세원,박병규 대한내분비학회 1990 Endocrinology and metabolism Vol.5 No.1

The authors observed the growth hormone (GH) response to pharmacologic agents (arginine-insulin, L-dopa) and growth hormone releasing hormone (GHRH) at Seoul National University Children's Hospital from September 1988 to August 1989. 42 patients included in the study were divided into 3 groups according to the peak plasma GH levels after pharmacologic stimulation test: complete GH deficiency group (25 patients), GH$lt;5 ng/ml; imcomplete GH deficiency group (6 patients), 5≤GH$lt;10 ng/ml; normal GH group (11 patients), GH≥10 ng/ml (GH levels of more than 10 ng/ml were measured from 8 patients). The peak plasma GH levels were measured from each group after GHRH stimulation test. The results were as follows: 1) The mean peak plasma levels of GH measured from complete and incomplete GH deficiency groups after GHRH stimulation test were higher than those after pharmacologic stimulation test, but were not different significantly in normal GH group. Following GHRH administration, peak plasma GH levels of more than 10 ng/mwere measured from 8 patients with idiopathic GH deficiency. Thus hypothalamic defect was suggested as the origin of their GH deficiency. 2) The serial mean plasma GH levels measured from normal GH group were persistently higher than those from complete and incomplete GH deficiency groups. But considerable overlapping of peak GH levels was observed among three groups. 3) The peak plasma GH levels after GHRH stimulation test were negatively correlated with age in complete GH deficiency and normal GH group. 4) The peak value of sequential mean plasma GH levels measured from incomplete GH deficiency group after GHRH stimulation test was reached earlier than that from complete GH deficiency group. In conclusion, GHRH stimulation test might be useful in the evaluation of readily releasable GH reservoir and speculation of putative mechanism of GH deficiency. But the discriminination of GH deficiency from normal GH status couldn't be made solely on the basis of the peak plasma GH levels after single GHRH stimulation test. Further study using different doses of GHRH is recommended for the evaluation of GH status (J. Kor Soc Endocrinol 5:14~22, 1990).

급성 피부점막 임파절 증후군 환자에서의 T림프구 및 그 아형의 변화와 감마글로블린 치료의 효과

김정애,오영균,박병규,윤희상,염명걸,김정수 의과학연구소 1990 全北醫大論文集 Vol.14 No.4

급성 피부점막 임파절 증후군 환아 13명을 대상으로 급성기와 아급성기에 T 림프구 및 그 아형의 변화와 혈청 감마마글로블린의 변화에 대해서 조사하고 아울러 MCLS 환아 42명을 대상으로 심혈관과 연관된 합병증을 심전 도 및 심에코드를 이용해서 조사하여 다음과 같은 결론을 얻었다. 1. 총 T 림프구(T1)과 T4/T8 비는 급성 기 및 아급성기에 모두 대조군에 비해서 유의한 감소가 있었으나 급성기과 아급성기 사이에는 변화가 없었다. 2.조력 T 림프구 (T4)는 급성기에 대조군에 비해서 유의한 감소가 있었으나 아급ㅁ성기에는 대조군과 차이가 없었다. 3.억제 T 림프구(T8)는 급성기에 대조군과 차이가 없다가 아급성기에는 유의안 증가가 있었다. 4.IgG, IgA, IgM치는 급성기에는 변화가 없다가 아급성기에는 현저한 증가를 보였다. 5.관상 동맥류는 전체 환자의 38%에서 생겼으며 제6병일에서 13병일 사이에서 (평균 :8.8±2.1일)관찰 할 수 있었다. 이상의 결과에 의하면 MCLS의 진행 과정 중 관상 동맥류의 형성은 대부분이 급성기에 이루어지며 이것은 조력 T 림프구(T4)의 감소로 인한 면역기능의 저하와 연관이 있을 것으로 보인다.

자가 말초혈 조혈모세포이식으로 장기간 관해를 보인 영아의 가족성 혈구탐식성 림프조직구증

김점수,서지현,황지영,김용숙,박찬후,김선주,맹국영,박병규 大韓血液學會 2001 大韓血液學會誌 Vol.36 No.4