• Asian-Australasian Journal of Animal Sciences
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• 제21권10호
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• pp.1404-1410
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• 2008

This study was performed to clarify whether the variation of stress related heat shock protein 70 (HSP70) (GenBank X68213) gene was associated with the nuclear morphological change of in vitro maturation and in vitro capacitation in oocytes of pig ovaries obtained at the slaughterhouse. The nucleic acid substitution of C to G at the 483rd position was found out in HSP70 K1 (290-512) from X68213. The ovaries were categorized into CC, CG, and GG genotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BsiHKA I). After the second in vitro maturation of immature fresh oocytes, the relation of nuclear morphological change in oocytes with the genotype of HSP70 K1 gene was such that the MII ratios of the genotype GG and CG (46.93% and 42.20%, respectively) were significantly higher than that of the CC genotype (10.71%) (p<0.05). With respect to in vitro maturation of frozen-thawed oocytes by an open pulled straw (OPS) method, the percentage of oocytes matured to MII stage of the CG genotype showed a higher trend than CC and GG genotypes. After the in vitro maturation of immature fresh oocytes and frozen-thawed oocytes by the OPS method, the relation of the pronuclei change in oocytes matured in vitro with HSP70 genotype was assessed, and the result showed that the enlarged sperm heads (ESH) of matured fresh oocytes and frozen-thawed oocytes were 80.0% and 60.0% in the CC genotype, respectively. The CC genotype group had a significantly higher rate of ESH than the CG and the GG genotype group (p<0.05). The ratios of polyspermic invasion were not different among HSP70 of the three genotypes. It was considered that the rate of in vitro maturation of fertilized oocytes was expected to differ according to genotype of the stress related gene.

• Tuberculosis and Respiratory Diseases
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• 제43권2호
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• pp.128-137
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• 1996

연구배경: 폐결핵은 고전적인 방법인 객담 도말 검사 및 배양검사로 전단할 수 있지만 객담 도말 민감도가 낮고 배양검사는 시일이 오래걸리는 단점이 있고 효과적인 객담배출이 되지 않는 환자에서는 검사가 불가능하기도 하다. 또한 최근에 개발된 PCR법은 신속하기는 하지만 위양성과 위음성이 문제가 되고 있다. 그래서 기관지내시경을 통한 기관지폐포세척액을 이용하여 세균학적 검사 및 PCR을 시행하여 폐결핵진단에 있어서 민감도와 예민도를 높이고 보다 신속한 진단을 할 수 있는지 연구하였다. 방법: 폐결핵 67예와 폐결핵이외의 폐질환을 가진 43예를 대상으로 객담 도말검사 및 배양검사를 시행하고, 기관지내시경을 시행하여 흉부 X-선상 병변이 보이는 엽상기관지에 기관지내시경을 위치하고 생리식염수 5cc로 3-5회에 걸쳐서 세척을 시행하여 세척액을 채취한 후 세균학적검사 및 PCR을 시행하였다. 결과: 1) 폐결핵환자의 기관지폐포세척액 결핵균 도말 검사 및 배양검사의 민감도는 각각 47.8% 및 80.6%로서, 객담 도말검사 및 배양검사의 민감도인 32.8% 및 57.4%보다 유의하게 높았다. 2) 폐결핵환자의 기관지폐포세척액 PCR의 민감도는 80.6%로서 배양검사의 민감도와 같았고, PCR의 배양검사에 대한 양성예측율은 81.5%였다. 3) 객담 도말검사 및 배양검사상 음성을 보인 폐결핵환자에서 시행한 기관지내시경을 통한 기관지폐포세척액 도말검사의 민감도는 23.1%였으며, 배양검사의 민감도는 100%였고, PCR의 민감도는 88.5%였으며, PCR의 배양검사에 대한 양성예측율 82.4%였다. 4) 기관지폐포세척액 PCR의 특이도는 77.0%였다. 5) 폐결핵의 최초 진단 후 치료시작까지의 기간은 객담 도말검사로 최초 진단이 된 경우가 평균 5일로 가장 빨랐고, 기관지폐포세척액 도말검사가 평균 9일, 기관지폐포세척액 PCR이 평균 26일이었으며, 객담배양검사는 평균 32일이었고, 기관지 폐포세척액 배양검사는 평균 56일로 가장 길었다. 결론: 폐결핵환자에서 기관지내시경을 통한 기관지폐포세척액의 결핵균 도말검사 및 배양검사는 객담을 이용한 검사보다 민감도가 높다. 그러므로, 객담을 배출하지 못하는 환자나 객담의 세균학적 검사가 음성인 환자에서 기관지내시경이 고려되어야 하며, 기관지폐포세척액의 PCR법까지 병행할 때에는 보다 신속하고 높은 진단율을 나타내므로, 조기에 적절한 항결핵제 투여에 도움이 될 것으로 기대된다.

• Clinical and Experimental Reproductive Medicine
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• 제33권2호
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• pp.85-95
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• 2006

목 적: CYP1B1은 자궁내막증의 발생과 관련된 에스트로겐의 대사 및 dioxin의 대사 모두와 관련이 있는 유전자로 생각하여 자궁내막증의 발생위험과 CYP1B1의 codon 119G${\rightarrow}$T, 432G${\rightarrow}$C, 449T${\rightarrow}$C, 453A${\rightarrow}$G의 유전자 다형성과의 관계에 대해서 알아보고자 하였다. 연구방법: 병리조직학적으로 자궁내막증 III기와 IV기임을 확인한 여성 199명과 대조군으로 자궁내막증 환자군과 연령이 비슷한 여성에서 양성 난소 낭종으로 수술을 시행하여 자궁내막증이 없음을 육안으로 확인한 183명을 대상으로 PCR 및 RFLP를 시행하여 CYP1B1의 codon 119G${\rightarrow}$T, 432C${\rightarrow}$G, 449T${\rightarrow}$C, 453A${\rightarrow}$G의 다형성을 조사하였다. 결 과: CYP1B1에서 $Ala^{l19}Ser$, $Va1^{432}Leu$, $Asn^{449}(T^{449}{\rightarrow}C)$, $Asn^{453}Ser$ 각각의 유전자 다형성은 그 분포와 위험도에 있어서 자궁내막증 환자와 대조군 간에 통계적으로 유의한 차이는 없었다. CYP1B1에서 앞서 밝힌 4가지 codon의 다형성을 조합한 결과 GG/CC/CC/AA의 유전자형을 갖는 여성에 비해 GG/GC+GG/TC+TT/AA의 유전자형을 갖는 경우만이 자궁내막증의 위험도가 2.056 (95% CI: 1.003-4.216)으로 유의하게 높은 것을 알 수 있었다. 결 론: 이상으로 볼 때 한국인 여성에서 중증자궁내막증 발생은 CYP1B1의 유전적 다형성과 관련이 있는 것으로 보이며, 향후 자궁내막증의 발생 기전을 밝히는데 주요한 자료가 될 것으로 본다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3403-3408
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• 2012

The molecular mechanisms involved in the progression of clear cell renal cell carcinomas (ccRCCs) are still unclear. The aim of this study was to analyse the relationships between expression of RALYL and clinical characteristics. In 41 paired samples of ccRCCs and adjacent normal tissues, we used real-time qPCR to evaluate the expression of RALYL mRNA. RALYL protein levels were determined in 146 samples of ccRCC and 37 adjacent normal tissues by immunohistochemistry. Statistical analysis was used to explore the relationships between expression of RALYL and the clinical characteristics (gender, age, tumor size, T stage, N stage, M stage, survival times and survival outcome) in ccRCC. In addition, these patients were follow-up period 64 months (range: 4~116months) to investigate the influence on prognosis. We found significantly differences between ccRCC tissues and normal tissues (p<0.001, paired-sample t test) in mRNA levels of RALYL. Immunohistochemistry analyses in 146 ccRCC samples and 37 adjacent normal tissues showed significantly lower RALYL protein levels in ccRCC samples (${\chi}^2$-test, p<0.001), inversely correlating with tumour size (p=0.024), T stage (0.005), N stage (p<0.001) as well as M stage (p=0.019), but not age (p=0.357) and gender (p=0.348). Kaplan-Meier survival analysis demonstrated that people with lower level of RALYL expression had a poorer survival rate than those with a higher level of RALYL expression, significantly different by the log-rank test (p=0.011). Cox regression analysis indicated that RALYL expression (p=0.039), N stage (p=0.008) and distant metastasis (p<0.001) were independent prognosis factors for the overall survival of ccRCC patients. We demonstrated that the expression of RALYL was significantly low in ccRCC and correlated with a poor prognosis in a large number of clinical samples. Our findings showed that RALYL may be a potential therapeutic target as well as a poor prognostic factor.

• Journal of Animal Science and Technology
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• 제46권3호
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• pp.295-300
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• 2004

본 연구는 도체성적을 보유하고 있는 제 31차, 32차 한우 후보종모우 집단 228두를 선발하여 DNA를 분리 정제 후 exon 2에 위치한 bovine leptin gene 염기서열 가운데 특정 염기서열을 갖는 2좌위의 primer를 합성하여 PCR 수행 후 PCR-product를 이용하여 2 종류의 제한효소 Kpn 2 I, Msp I 으로 반응시킨 후 두 가지 형태의 대립 유전자를 검정하여 경제형질과의 관련성을 분석하였다. PCR-RFLP를 통하여 얻어진 leptin gene의 유전자형 빈도는 Kpn2 I의 경우 C 유전자 빈도(0.25)보다 T 유전자 빈도(0.75)가 높게 나타났으며 Msp I으로 처리한 경우 M 유전자 빈도(0.35)보다 m 유전자 빈도(0.65)가 높게 나타났다. 통계적 분석을 통하여 각 유전자형에 대한 경제형질과의 관련성을 분석한 결과 제한효소 Kpn2 I으로 처리한 경우 도체율에서 CT 유전자형과 CC 유전자형 사이에 유의적 차이가 나타났으며(P < 0.05), Msp I의 경우 도체중 Mm 유전자형과 mm 유전자형 사이에 통계적 유의성이 나타났다(P < 0.05).

• 생명과학회지
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• 제24권5호
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• pp.567-572
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• 2014

초롱꽃과에 속하는 도라지는 항염증, 항산화, 항암작용뿐 아니라 간기능 강화에도 작용하여 한방에서 만성염증성 질환의 치료제로 널리 사용되고 있다. 이번 연구에서 마우스 대식세포주인 RAW 246.7 세포와 비장으로부터 분리한 T세포에서 도라지 추출액(CC), 도라지 추출물이 함유된 활맥단 추출액(HWAL, 도라지 82% 함유) 및 맥환추출액(MAEK, 도라지 7% 함유)을 처리하여 대식세포와 T세포의 활성과 관련 있는 유전자의 발현을 통해 면역증진 및 항염증 효과를 real-time RT-PCR 기법을 통해 확인해 보았다. 그 결과, 활성화된 T세포에서 많이 발현되는 c-fos (10배 이상), CD40L 유전자(6배 이상)의 경우, 맥환 추출액 처리군에서 가장 높은 발현이 유도되어 면역활성 효과가 높을 것으로 사료되었다. 도라지 추출액과 활맥단 추출액의 경우도 c-fos 유전자의 발현을 유의적으로 증가시켰지만 미약하였고, CD40L의 경우는 영향을 주지 않았다. 도라지 추출액을 처리한 군에서 면역활성을 유도하는 M1 대식세포에서 많이 발현되는 iNOS 유전자의 발현이 유의하게 증가되었다. 반면에 활맥단 추출액을 처리한 대식세포는 M2 표식인자인 Ym1, arginase1 (ARG1) 유전자의 발현이 유의적으로 상승하였다. 결론적으로 도라지가 다량 포함된 맥환 추출액은 면역활성을 유도하는데 탁월한 효과를 나타내며, 도라지 함유가 적게 포함된 활맥단 추출액은 항염증 작용에 탁월한 효과가 있을 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제54권6호
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• pp.703-710
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• 2016

The trichomonad species Tritrichomonas foetus and Pentatrichomonas hominis were recently detected in the feces of dogs with diarrhea. However, little information is available on the prevalence and pathogenicity of these parasites in the canine population. Therefore, the aim of this study was to determine the prevalence and molecular characterization of trichomonads infecting pet dogs in Anhui and Zhejiang provinces, east China. In total, 315 pet dogs, with or without diarrhea, from 7 pet hospitals were included in this epidemiological survey. Microscopy and PCR detected P. hominis in 19.7% (62/315) and 31.4% (99/315) of fecal samples, respectively. T. foetus infection was detected in 0% (0/315) of samples with microscopy and in 0.6% (2/315) with PCR. The prevalence of P. hominis was significantly higher in young dogs (${\leq}12months$) than in adult dogs (>12 months), and was significantly higher in diarrheic dogs (50.6%) than in non-diarrheic dogs (24.3%; P<0.05). Infection with T. foetus did not correlate with any risk factors evaluated in this study. A sequence analysis of the P. hominis PCR products showed minor allelic variations between our sequences and those of P. hominis strains from other hosts in different parts of the world. Type CC1 was the most common strain in dogs in east China. The internal transcribed spacer 1 (ITS1)-5.8S rRNA gene sequences from the 2 T. foetus isolates detected in this study displayed 100% identity and were homologous to the sequences of other strains isolated from domestic cats in other countries.

• 동의생리병리학회지
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• 제25권4호
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• pp.663-668
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• 2011

Galgeun-tang (GGT) has been a great source for treating cold diseases in the folk medicine recipe. Carbon tetrachloride ($CCl_4$) is one type of hepatotoxin that can eventually cause liver injury. During the experiment, we first studied the protective effects of GGT against $CC_4$-induced hepatotoxicity. GGT was pretreated for 3 h, and 1% $CCl_4$ was added to mouse primary liver cells. After 4 h, ROS generation and expression of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx)) were analyzed by FACS and real time PCR. Also, the activities of ALT and LDH were measured using cultured medium. The hepatic levels of TNF-alpha and iNOS, which are related to inflammation and stress response gene, HSP72 and HO-1 were analyzed by PCR or real time PCR. Liver tissues were analyzed by HE stain. From the observation, we discovered that GGT treatment protects $CCl_4$-induced hepatotoxicity, and that GGT pretreatment decreases ROS generation, TNF-alpha and iNOS expression. However, gene expression of CAT, SOD, GPx, HSP72 and HO-1 were increased by GGT. These results lead to the conclusion that GGT has protective effects against $CCl_4$-induced hepatotoxicity.

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.77-82
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• 2014

The study aimed to evaluate the potential effects of feeding with phytase transgenic corn (PTC) on organ weight, serum biochemical parameters and nutrient digestibility, and to determine the fate of the transgenic DNA in laying hens. A total of 144 50-week-old laying hens were grouped randomly into 2 treatments, with 8 replicates per treatment and 9 hens per replicate. Each treatment group of hens was fed with diets containing 62.4% non-transgenic conventional corn (CC) or PTC for 16 weeks. The phytase activity for CC was 37 FTU/kg of DM, whereas the phytase activity for PTC was 8,980 FTU/kg of DM. We observed that feeding PTC to laying hens had no adverse effect on organ weight or serum biochemical parameters (p>0.05). A fragment of a poultry-specific ovalbumin gene (ov) was amplified from all tissues of hens showing that the DNA preparations were amenable to PCR amplification. Neither the corn-specific invertase gene (ivr) nor the transgenic phyA2 gene was detected in the breast muscle, leg muscle, ovary, oviduct and eggs. The digestibility data revealed no significant differences between the hens that received the CC- and PTC-based diets in the digestibility of DM, energy, nitrogen and calcium (p>0.05). Phosphorus digestibility of hens fed the PTC-based diet was greater than that of hens fed the CC-based diet (58.03% vs 47.42%, p<0.01). Based on these results, it was concluded that the PTC had no deleterious effects on the organ weight or serum biochemical parameters of the laying hens. No recombinant phyA2 gene was detected in muscle tissues and reproductive organs of laying hens. The novel plant phytase was efficacious in improving the phosphorus digestibility of laying hens.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.215-222
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• 2002

Objective : Previous studies have suggested that hyperhomocysteinemia and methylenetetrahydrofolate reductase (MTHFR C677T) mutations are associated with increased risk of recurrent spontaneous abortion (RSA). Recently, a second site polymorphism in MTHFR, 1298A-->C, which changes a glutamic acid into an alanine residue, was shown to be associated with a decreased enzyme activity. We tested whether the variant alleles of MTHFR C677T and A1298C are risk factor (biomarker) for RSA. Materials and Methods: We analyzed DNA from a case-control study in the Korean DNA was extracted from blood samples of 118 patients with RSA and 123 healthy fertile patients as the controls. MTHFR variant alleles were determined by a PCR-restriction fragment length polymorphism assay. Results: We found no evidence for an association between 677TT genotype and risk of RSA (OR=1.95, 95% CI=$0.84{\sim}4.50$, p=0.12). However, the MTHFR 1298AC (OR=0.36, 95% CI=$0.20{\sim}0.63$, p=0.0004) and 1298AC+CC (OR=0.35, 95% CI=$0.20{\sim}0.61$, p=0.0002) genotypes were lower among 118 RSA cases compared with 123 controls, conferring a 2.8-fold decrease in risk of RSA, respectively. Moreover, the combined genotypes of MTHFR 677CC/1298AC (OR=0.30, 95% CI=$0.10{\sim}0.88$, p=0.029) and 677CT/1298AC (OR=0.77, 95% CI=$0.60{\sim}0.99$, p=0.043) also showed significantly lower risk than those with MTHFR 677CC/1298AA type. Conclusion: MTHFR 1298AC, MTHFR 677CC/1298AC and 677CT/1298AC genotypes may represent genetic markers for the protection of RSA at least in Korean women.