Guangdong Provincial Key laboratory of Bioengineering Medicine

Guangzhou, China

Guangdong Provincial Key laboratory of Bioengineering Medicine

Guangzhou, China
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Yuan F.-M.,Jinan University | Guo X.-Y.,Guangdong Provincial Key Laboratory of Bioengineering Medicine
Chinese Journal of Pharmaceutical Biotechnology | Year: 2014

Transient Gene Expression (TGE) is a fast approach for r-protein production in the absence of selection of high-expression cell line. There are few reports about TGE in insect cell although insect cell line SF9 is a widely used in gene expression system. In this study, the factors influencing transfection efficiency, including amount of DNA, ratio of DNA and PEI, and cell density were optimized to express eGFP and TNFR-Fc in insect cell Sf9 in suspension. Under the optimized conditions, 37% of the transfection efficiency and 700 μg/L recombinant TNFR-Fc were obtained. © 2014, Editorial Board of Pharmaceutical Biotechnology. All right reserved.


Xie Q.-L.,Jinan University | Xie Q.-L.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Wang Y.-Y.,Jinan University | Wang Y.-Y.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | And 6 more authors.
Chinese Journal of Pharmaceutical Biotechnology | Year: 2013

To clone the shark Troponin I (TnI) from whitespotted bambooshark (Chiloscyllium plagiosum Bonnet) and to express in E. coli. as well as to identify the anti-angiogenesis ability of this protein, the gene of human Troponin I (TnI) homolog from liver of whitespotted bambooshark (Chiloscyllium plagiosum Bonnet) was cloned, followed by recombinant expression in E. coli. Then the inhibitory ability of shark TnI for angiogenesis was investigated by MTT method and chicken embryo allantoic membrane (CAM) assessment. Results showed that the cDNA of shark TnI was 692 bp in length, with an ORF of 552 bp (encoding a polypeptide of 183 amino acids). It has the highest amino acid identity (85.2%) with dogfish shark TnI, whereas 68.9%, 64.5% and 62.3% with TnIs of human, salmon and zebrafish, respectively. The recombinant shark TnI was found to inhibit the proliferation of human umbilical vein endothelial cells (HUVECs) in a dose-dependent fashion and angiogensis of chicken embryo allantoic membrane. The TnI of whitespotted bambooshark (Chiloscyllium plagiosum Bonnet) was confirmed to inhibit angiogenesis.


Wang H.,South China University of Technology | Zhu W.,South China University of Technology | Huang Y.,Jinan University | Li Z.,South China University of Technology | And 3 more authors.
Acta Biomaterialia | Year: 2016

Nano-drug delivery systems that integrate inorganic and organic or even bioactive components into a single nanoscale platform are playing a greatly important role in cancer treatment. Here, the fabrication of a versatile nanocarrier based on self-assembled structures of gold nanoparticles (AuNPs)-zein is reported, which displays high drug-loading efficiency for needle-shaped hydroxycamptothecin (HCPT) nanocrystals. The surface modification with folate-conjugated polydopamine (PFA) renders them stable and also facilitates their selective cellular internalization and enhancement of endocytosis. The release of payloads from nanocomplexes (NCs) was shown to be limited at physiological pH (17.1. ±. 2.8%) but significantly elevated at endosomal/lysosomal pH (58.4. ±. 3.0%) and at enzymatic environment (81.4. ±. 4.2%). Compared to free HCPT and its non-targeting equivalent, HCPT@AuNPs-Zein-PFA, exerted a superior tumor suppression capacity as well as low side effects due to their active and passive targeting delivery both in vitro and in vivo. These results suggest that the NCs with well-defined core@shell nanostructures encapsulated with HCPT nanocrystals hold great promise to improve cancer therapy with a high efficiency in the clinic. Statement of significance: A novel nanocomplex encapsulated HCPT nanocrystals was designed to achieve selective cellular uptake by endocytosis, acid responsive release in the tumor microenvironment and excellent tumor suppression without toxicity. This nanocomplex with conjugation of folate was stable in the bloodstream, with minimal drug release in extracellular conditions, leading to prolonged blood circulation and high accumulation in tumor tissues. The entrapment of nanocrystal drug into nanomaterials might be capable of delivering drugs in a predictable and controllable manner. © 2017 Acta Materialia Inc.


Zhang S.,Jinan University | Zhang S.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Xing K.,Jinan University | Xing K.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | And 6 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2011

The experiment was conducted by directed evolution strategy (error-prone PCR) to improve the activity of aflatoxin detoxifzyme with the high-throughput horse radish peroxidas and recessive brilliant green (HRP-RBG) screening system. We built up a mutant library to the order of 104. Two rounds of EP-PCR and HRP-RBG screening were used to obtain three optimum mutant strains A1773, A1476 and A2863. We found that mutant A1773 had upper temperature tolerance of 70°C and that its enzyme activity was 6.5 times higher than that of the parent strain. Mutant strains A1476 worked well at pH 4.0 and its enzyme activity was 21 times higher than that of the parent strain. Mutant A2863 worked well at pH 4.0 and pH 7.5, and its enzyme activity was 12.6 times higher than that of the parent strain. With DNA sequencing we found that mutant A1773 revealed two amino acid substitutions, Glu127Lys and Gln613Arg. Mutant A1476 revealed four amino acid substitutions: Ser46Pro, Lys221Gln, Ile307Leu and Asn471Ile. Mutant A2863 revealed four amino acid substitutions: Gly73Ser, Ile307Leu, Val596Ala and Gln613Arg. The results provided a useful illustration for the deep understanding of the relationship between the function and structure of aflatoxin detoxifzyme. © 2011 CJB All rights reserved.


Xie Q.,Jinan University | Xie Q.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Xinyong G.,Jinan University | Xinyong G.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | And 4 more authors.
Cytotechnology | Year: 2013

Polyethylenimine (PEI) has been used widely in transient gene expression studies of mammalian cells. We performed transient gene expression in suspension Chinese hamster ovary cells using a one-step transfection procedure in which DNA and PEI were simultaneously added to a cell culture in suspension without prior PEI/DNA complex incubation. To further understand the effect of PEI/DNA formation on the transfection and expression of exogenous gene in shaking state, we investigated the diameter and overcharge of the PEI/DNA complex. The results showed that the diameter of the complex was smaller with more positive charge when the PEI/DNA ratio was higher. Moreover, DNA more easily penetrated cells and nuclei at higher PEI concentrations. The highest transcription level, transfection efficiency, and GFP expression were obtained when the PEI/DNA ratio was 5:1. © 2012 Springer Science+Business Media B.V.


Chen X.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Lu J.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Ji Y.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Hong A.,Guangdong Provincial Key Laboratory of Bioengineering Medicine | Xie Q.,Guangdong Provincial Key Laboratory of Bioengineering Medicine
Tumor Biology | Year: 2014

Bone is one of the most common metastatic sites for breast cancer. In this study, we observed a promoting effect of osteoblast-conditioned medium (OCM) on the migration of MCF-7, a noninvasive cell line of breast cancer cells. Cytokine antibody array was used to compare the cytokines of OCM with the conditioned medium of non-differentiated osteoblast cells, which consequently revealed factors related to migration, such as IL8, IL6, CSF2 (G-CSF), CSF3 (GMCSF), and TNFRSF11B (osteoprotegerin). The expression of genes related to migration was also estimated with a PCR array, which showed that 9 genes were upregulated and 26 genes downregulated. Moreover, activated p38, ERK, and AKT pathways were found in the OCMtreatment group. This finding indicated the migration ability of breast cancer cells, which move toward the bone depending on the presence of specific cytokines in its surrounding microenvironment.


Zhou T.,Jinan University | Zhou T.,Guangdong Provincial Key laboratory of Bioengineering Medicine | Zhu X.,Jinan University | Su J.,Jinan University | And 3 more authors.
Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | Year: 2012

Activity losing during the covalent immobilization of enzyme is a serious problem. Here we studied organic phase immobilization by using glucose oxidase (GOD) as a model. After lyophilized at optimum pH, GOD is covalently immobilized onto glutaraldhyde-activated chitosan microsphere carrier under the condition of water, 1, 4-dioxane, ether and ethanol separately. The special activities, enzyme characterization and kinetic parameters are determined. Results show that all of the organic phase immobilized GODs have higher special activities and larger Kcat than that of aqueous phase. Under the conditions of 0.1% of glutaraldehyde,1.6% moisture content with 80 mg of GOD added to per gram of carrier, 2.9-fold of the special activity and 3-fold of the effective activity recovery ratio were obtained, and 3-fold of the residue activity was demonstrated after 7 runs when compares 1, 4-dioxane phase immobilized GOD with water phase immobilized one. In addition, kinetic study shows that 1, 4-dioxane immobilized GOD (Km app=5.63 mmol/L, Vmax=1.70 μmol/(min·mg GOD), Kcat=0.304 s-1) was superior to water immobilized GOD (Km app=7.33 mmol/L, Vmax=1.02 μmol/(min·mg GOD), Kcat=0.221 s-1). All above indicated GOD immobilized in proper organic media presented a better activity with improved catalytic performance. Organic phase immobilization might be one of the ways to overcome the conformational denature of enzyme protein during covalent modification.


PubMed | Guangdong Provincial Key Laboratory of Bioengineering Medicine
Type: Journal Article | Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine | Year: 2014

Bone is one of the most common metastatic sites for breast cancer. In this study, we observed a promoting effect of osteoblast-conditioned medium (OCM) on the migration of MCF-7, a noninvasive cell line of breast cancer cells. Cytokine antibody array was used to compare the cytokines of OCM with the conditioned medium of non-differentiated osteoblast cells, which consequently revealed factors related to migration, such as IL8, IL6, CSF2 (G-CSF), CSF3 (GM-CSF), and TNFRSF11B (osteoprotegerin). The expression of genes related to migration was also estimated with a PCR array, which showed that 9 genes were upregulated and 26 genes downregulated. Moreover, activated p38, ERK, and AKT pathways were found in the OCM treatment group. This finding indicated the migration ability of breast cancer cells, which move toward the bone depending on the presence of specific cytokines in its surrounding microenvironment.

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