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Shenglin Hu,Dongmei Wang,Jiong Hong 한국생물공학회 2018 Biotechnology and Bioprocess Engineering Vol.23 No.1
In this study, a simple, inexpensive and fast β-glucosidase immobilization system was constructed and evaluated in isoflavone glycosides hydrolysis. A β-glucosidase gene from Thermoascus aurantiacus IFO9748 was recombinantly expressed in Pichia pastoris KM71H and immobilized on regenerated amorphous cellulose (RAC) by fused cellulose binding module 3. Through simple mixing cellulose and crude enzyme for 15 min under room temperature, 96.04% β-glucosidase was immobilized onto RAC. The optimum temperature for β-glucosidase activity was increased by 5ºC after immobilization. The half-life (t½) of heat inactivation of immobilized enzyme at 60oC was improved over 8 folds. After 30 rounds recycled at 40oC, 96.9% daidzin and 98.9% genistin could still be hydrolyzed. A continuous hydrolysis system was also constructed, and at the flow rate of 0.2 mL/min after 30 h hydrolysis, 95.6% genistin and 90.2% daidzin can still be hydrolyzed. Combined the simple and high efficient enzyme immobilization procedure and inexpensive cellulose, this scalable and practical system may have broad prospects for industrial utilization.
miR-182-5p Inhibits NKAPL Expression and Promotes the Proliferation of Osteosarcoma
Shen Yang,Kaixi Chen,Kun Cao,Shenglin Xu,Chengxiao Ma,Yongping Cai,Yong Hu,Yejin Zhou 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.5
Purpose Osteosarcoma, a malignant bone tumor, has the lowest survival rate among all pediatric cancers. NF-κB-activating protein-like (NKAPL) is highly homologous with NKAP. The expression of NKAPL is downregulated in primary liver cancer and breast cancer, and plays a role of tumor suppressor gene. However, the role of NKAPL in osteosarcoma has not been reported. Materials and Methods We explored the effect of NKAPL on the proliferation of osteosarcoma cells by immunohistochemical, RT-PCR, Western blot, and double luciferase reporter gene analysis. Results The low expression of NKAPL mRNA was correlated with distant metastasis (P = 0.017), tumor size (P = 0.023), and clinical stage (P < 0.001). The NKAPL expression level in MG63 and U2OS cells was lower than that in Nhost cells. Downregulation of NKAPL expression in Nhost cells could promote cell proliferation and upregulation of NKAPL expression in MG63 and U2OS cells could inhibit cell proliferation. miR-182-5p expression was negatively correlated with NKAPL mRNA expression (R2 = 0.1169, P = 0.0099). After upregulating NKAPL expression, the Notch1, hes1, hey2, and cyclin D1 expression levels were significantly decreased, with G0/G1 phase arrest and G2/M phase reduction. Conclusions miR-182-5p targeted NKAPL and inhibit NKAPL expression in osteosarcoma. miR-182- 5p could regulate cell cycle and promote tumor proliferation through upregulating Notch signaling pathway.
Aberrant microRNAs Expression in CD133+/CD326+ Human Lung Adenocarcinoma Initiating Cells from A549
Sheng Lin,Zheng-tang Chen,Jian-guo Sun,Jing-bo Wu,Hai-xia Long,Cong-hui Zhu,Tong Xiang,Hu Ma,Zhong-quan Zhao,Quan Yao,An-mei Zhang,Bo Zhu 한국분자세포생물학회 2012 Molecules and cells Vol.33 No.3
Increasing evidence demonstrates that miRNAs are in-volved in the dysregulation of tumor initiating cells (TICs) in various tumors. Due to a lack of definitive markers, cell sorting is not an ideal separation method for lung adeno-carcinoma initiating cells. In this study, we combined pa-clitaxel with serum-free medium cultivation (inverse-induc-tion) to enrich TICs from A549 cells, marked by CD133/ CD326, defined features of stemness. We next investigated aberrant microRNAs in this subpopulation compared to normal cells with miRNA microarray and found that 50 miRNAs exhibited a greater than 2-fold change in expres-sion. As further validation, 10 miRNAs were chosen to perform quantitative RT-PCR on the A549 cell line and primary samples. The results suggest that aberrant ex-pression of miRNAs such as miR-29ab, miR-183, miR-17-5p and miR-127-3P may play an important role in regulat-ing the bio-behavior of TICs.