소나무재선충(pine wood nematode, PWN) 감염 소나무를 현장에서 신속하게 진단을 할 수 있는 방법은 현재 없다. 본 연구에서는 PWN특이적인 항원으로 알려진 PWN-GaLectin을 baculovirus발현체계로 발현시켜서 총 1,464개의 fusion hybridoma 세포 주 라이브러리 를 제작 하는 항원으로 이용하였다. 총 1,464개의 fusion hybridoma 세포 주 중, PWN-GaLectin에 대한 높은 반응을 보이는 62개의 fusion hybridoma 세포 주를 선별했다. 이들 중, 표준 PWN 감염소나무 PBS추출물과 PWN 단백질 추출물에 강한 반응을 보이는 세포 주 12개를 선 별하여 단클론 항체(monoclonal antibody, Mab) 분비세포 주 확립을 위한 limited dilution을 실시하였다. Mab분비세포 주 확립을 위해서 표 준 PWN 감염소나무 추출물에 대한 반응이 표준 정상 소나무 추출물 보다 높은 세포 주들을 선별했다. 그리고 추가로 PWN 단백질 추출물에 대 해서 3종의 비 병원성 선충 단백질 추출물 보다 높은 반응을 보이는 세포 주들도 선별, 확립했다. 본 연구에서 확립된 Mab들을 우리는 현장과 실 험실에서 사용할 수 있는 신속진단키트의 개발에 이용할 수 있을 것이다.
The purpose of this study was to investigate the effect of estrus synchronization to altrenogest regumate (progesterone), PMSG/hCG, and artificial insemination (AI) on galectin-3, FGF-9 gene and protein expression. The morpho-metrical parameters of the endometrium and the number of corpora lutea (CL) were recorded. RNA was isolated from endometrial, oviduct and ovary tissues of non-synchronized (Control; n = 7) and AI synchronized (regumate, PMSG/hCG; n = 7) sows. The total number of CL was higher (P<0.05) in pigs treated with regumate/PMSG/hCG. The content of gelactin-3 and FGF-9 mRNA in pre-embryonic development stages increased on particular days, in control and studied in regumate/PMSG/hCG administered pigs. Gelactin-3 and FGF-9 were affected by regumate/PMSG/hCG treatment in the both pre-embryonic development stages (P<0.001, P<0.05) and encdometrial tissue (P<0.001, P<0.01). The regumate/PMSG/hCG treatment resulted in elevated expression of gelactin-3 (P<0.001) and FGF-9 (P<0.005) in oviduct and ovary tissues in comparison to control sows. Moreover, oviduct amount of gelectin-3 mRNA was higher in regumate/PMSG/hCG sows in comparison to the control group (P<0.05), whereas, expression characteristics of gelactin-3 and FGF-9 were investigated by hematoxylin and eosin stained and immunohistochemical staining. The results showed that galectin-3 and FGF-9 were significantly shown in the endometrium, oviduct and ovary tissues of the regumate/PMSG/ hCG. Presented data show that exogenous hormones administration can affect gene and protein expression in the sow reproductive tract.
The purpose of this study was to examine the anti-metastatic effect of Juniperus chinensis extract by inhibition of galectin-3 expression. For this study, we carried out the experiment of galectin-3 expression assay and in vivo metastasis mouse model bearing colon 26-M3.1 lung carcinoma, and also MMP-1, MMP-2 and MMP-9 inhibitory assays relating metastasis mechanism. J. chinensis extract significantly showed the inhibitory effect of basal expression and MNNG-induced expression of galectin-3 in colon 26-M3.1 lung carcinoma at the all tested doses (5, 50 and 500 ㎍/㎖). In case of the inhibition of MMP-1, MMP-2 and MMP-9, the weak activity was showed at the doses of 50 and 500 ㎍/㎖ and was not detected at the dose of 5 ㎍/㎖. However, J. chinensis extract significantly showed the anti metastatic activity on mouse mode bearing colon 26-M3.1 lung carcinoma. Therefore, this study strongly suggests that J. chinensis is a potent inhibitor of galectin-3 expression, and holds great promise for use in inhibiting metastasis induced by elevated expression of galectin-3.