1.Cloning expression and purification of glycerol dehydrogenase from Klebsiella pneumoniae.
Tingting ZHANG ; Baishan FANG ; Geng WANG ; Feifei WANG
Chinese Journal of Biotechnology 2008;24(3):495-499
The gldA gene coding glycerol dehydrogenase (GDH) was amplified by PCR with the genomic DNA of Klebsiella pneumoniae as the template. The gldA were inserted in pMD-18T to construct the recombinant cloning vector pMD-gldA. After the DNA sequence was determined, the gldA was subcloned into expression vector pET-32a (+) to construct the recombinant expression vector pET-32gldA. Upon lactose induction, soluble GDH was over-produced by E. coli BL21 (DE3) harboring the expression construct. Recombinant GDH purified by Ni-NTA affinity chromatography showed a single band about 54 kD on SDS-PAGE gel, and the specified activity was about 188 u/mg, the purification fold is 3 times and the activity recovery is 67.5%.
Chromatography, Affinity
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Cloning, Molecular
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DNA, Bacterial
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genetics
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Escherichia coli
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genetics
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isolation & purification
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metabolism
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Klebsiella pneumoniae
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enzymology
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genetics
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Sugar Alcohol Dehydrogenases
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biosynthesis
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genetics
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isolation & purification
2.Analysis of 24 cases of cryptococcal meningitis treated with fluconazole
Xueting OU ; Changming GENG ; Bin XU ; Jiqin WU ; Xinyu WANG ; Shu CHEN ; Feifei YANG ; Wanqin ZHANG ; Liping ZHU ; Xinhua WENG
Chinese Journal of Infectious Diseases 2009;27(6):357-359
Objective To evaluate clinical features,therapeutic effects and outcomes of patients with non-human immunodeficiency virus(HIV)-infected cryptococcal meningitis treated with fluconazole or fluconazole and flucytosine.Methods Twenty-four cases of non-HIV-infected cryptococcal meningitis(fluconazole with or without flucytosine as initial therapy)in Huashan Hospital,Fudan University from 1997 to 2007 were retrospectively reviewed.Clinical manifestations,therapeutic effects and outcomes of the patients were collected.Results Fluconazole was administered with median dosage of 400 mg/d,for a median duration of 20.5 days.After fluconazole initial therapy for 2 weeks,16.7% showed partial response,83.3% showed no response,and the overall response rate was 16.7%.After 10 weeks,33.3% showed partial response,29.2% showed complete response,16.7% showed no response,and the overall response rate was 62.5%.Mortality at week 10 was 20.8%.Twenty-two patients who failed to respond to initial therapy were switched to other antifungal drugs(amphotericin B,amphotericin B colloidal dispersion,itraconazole)or other fluconazole containing combined therapy.Eleven out of the 24 patients died during one-year follow-up,8 of whom died of eryptococcal meningitis,and 3 died of other diseases.Conclusions The initial therapy of fluconazole with or without flucytosine is inefficient,and most of the patients need other antifungal drugs because of initial therapy failure.Therefore,fluconazole might not be appropriate for initial therapy in non-HIV-infected cryptococcal meningitis.
3. Proteomics analysis of hippocampus and striatum in rats with hyperglycemia using iTRAQ technique
Ablat DILMURAT ; Xinling YANG ; Yumin JIA ; Feifei GENG
Chinese Journal of Endocrinology and Metabolism 2019;35(11):973-980
Objective:
To screen the deferentially expressed proteins in hippocampus and striatum in rat models of diabetes mellitus and normal SD rats, and to elucidate the effects of hyperglycemia on central nervous system.
Methods:
SD rats were randomly divided into normal group(
4.Targeting ERK, an Achilles' Heel of the MAPK pathway, in cancer therapy.
Feifei LIU ; Xiaotong YANG ; Meiyu GENG ; Min HUANG
Acta Pharmaceutica Sinica B 2018;8(4):552-562
The mitogen-activated protein kinases (MAPK) pathway, often known as the RAS-RAF-MEK-ERK signal cascade, functions to transmit upstream signals to its downstream effectors to regulate physiological process such as cell proliferation, differentiation, survival and death. As the most frequently mutated signaling pathway in human cancer, targeting the MAPK pathway has long been considered a promising strategy for cancer therapy. Substantial efforts in the past decades have led to the clinical success of BRAF and MEK inhibitors. However, the clinical benefits of these inhibitors are compromised by the frequently occurring acquired resistance due to cancer heterogeneity and genomic instability. This review briefly introduces the key protein kinases involved in this pathway as well as their activation mechanisms. We also generalize the correlations between mutations of MAPK members and human cancers, followed by a summarization of progress made on the development of small molecule MAPK kinases inhibitors. In particular, this review highlights the potential advantages of ERK inhibitors in overcoming resistance to upstream targets and proposes that targeting ERK kinase may hold a promising prospect for cancer therapy.
5.mTORC2/RICTOR exerts differential levels of metabolic control in human embryonic, mesenchymal and neural stem cells.
Qun CHU ; Feifei LIU ; Yifang HE ; Xiaoyu JIANG ; Yusheng CAI ; Zeming WU ; Kaowen YAN ; Lingling GENG ; Yichen ZHANG ; Huyi FENG ; Kaixin ZHOU ; Si WANG ; Weiqi ZHANG ; Guang-Hui LIU ; Shuai MA ; Jing QU ; Moshi SONG
Protein & Cell 2022;13(9):676-682
6.Correction to: mTORC2/RICTOR exerts differential levels of metabolic control in human embryonic, mesenchymal and neural stem cells.
Qun CHU ; Feifei LIU ; Yifang HE ; Xiaoyu JIANG ; Yusheng CAI ; Zeming WU ; Kaowen YAN ; Lingling GENG ; Yichen ZHANG ; Huyi FENG ; Kaixin ZHOU ; Si WANG ; Weiqi ZHANG ; Guang-Hui LIU ; Shuai MA ; Jing QU ; Moshi SONG
Protein & Cell 2022;13(12):961-961
7.Structure-based drug discovery of novel fused-pyrazolone carboxamide derivatives as potent and selective AXL inhibitors.
Feifei FANG ; Yang DAI ; Hao WANG ; Yinchun JI ; Xuewu LIANG ; Xia PENG ; Jiyuan LI ; Yangrong ZHAO ; Chunpu LI ; Danyi WANG ; Yazhou LI ; Dong ZHANG ; Dan ZHANG ; Meiyu GENG ; Hong LIU ; Jing AI ; Yu ZHOU
Acta Pharmaceutica Sinica B 2023;13(12):4918-4933
As a novel and promising antitumor target, AXL plays an important role in tumor growth, metastasis, immunosuppression and drug resistance of various malignancies, which has attracted extensive research interest in recent years. In this study, by employing the structure-based drug design and bioisosterism strategies, we designed and synthesized in total 54 novel AXL inhibitors featuring a fused-pyrazolone carboxamide scaffold, of which up to 20 compounds exhibited excellent AXL kinase and BaF3/TEL-AXL cell viability inhibitions. Notably, compound 59 showed a desirable AXL kinase inhibitory activity (IC50: 3.5 nmol/L) as well as good kinase selectivity, and it effectively blocked the cellular AXL signaling. In turn, compound 59 could potently inhibit BaF3/TEL-AXL cell viability (IC50: 1.5 nmol/L) and significantly suppress GAS6/AXL-mediated cancer cell invasion, migration and wound healing at the nanomolar level. More importantly, compound 59 oral administration showed good pharmacokinetic profile and in vivo antitumor efficiency, in which we observed significant AXL phosphorylation suppression, and its antitumor efficacy at 20 mg/kg (qd) was comparable to that of BGB324 at 50 mg/kg (bid), the most advanced AXL inhibitor. Taken together, this work provided a valuable lead compound as a potential AXL inhibitor for the further antitumor drug development.