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Determining Osteogenic Differentiation Efficacy of Pluripotent Stem Cells by Telomerase Activity
Siqi Zhang,Yuhua Sun,Yi Sui,Yan Li,Zuyuan Luo,Xiao Xu,Ping Zhou,Shicheng Wei 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.6
BACKGROUND: Bone tissue engineering based on pluripotent stem cells (PSCs) is a new approach to deal with bone defects. Protocols have been developed to generate osteoblasts from PSCs. However, the low efficiency of this process is still an important issue that needs to be resolved. Many studies have aimed to improve efficiency, but developing accurate methods to determine efficacy is also critical. Studies using pluripotency to estimate efficacy are rare. Telomerase is highly associated with pluripotency. METHODS: We have described a quantitative method to measure telomerase activity, telomeric repeat elongation assay based on quartz crystal microbalance (QCM). To investigate whether this method could be used to determine the efficiency of in vitro osteogenic differentiation based on pluripotency, we measured the pluripotency pattern of cultures through stemness gene expression, proliferation ability and telomerase activity, measured by QCM. RESULTS: We showed that the pluripotency pattern determined by QCM was similar to the patterns of proliferation ability and gene expression, which showed a slight upregulation at the late stages, within the context of the general downregulation tendency during differentiation. Additionally, a comprehensive gene expression pattern covering nearly every stage of differentiation was identified. CONCLUSION: Therefore, this assay may be powerful tools for determining the efficiency of differentiation systems based on pluripotency. In this study, we not only introduce a new method for determining efficiency based on pluripotency, but also provide more information about the characteristics of osteogenic differentiation which help facilitate future development of more efficient protocols.
Siqi Li,Zhipeng Su,Chengjun Zhang,Zhuofei Xu,Xiaoping Chang,Jiawen Zhu,Ran Xiao,Lu Li,Rui Zhou 한국유전학회 2018 Genes & Genomics Vol.40 No.8
Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae has led to severe economic losses in the pig industry worldwide. A. pleuropneumoniae displays various levels of antimicrobial resistance, leading to the dire need to identify new drug targets. Protein–protein interaction (PPI) network can aid the identification of drug targets by discovering essential proteins during the life of bacteria. The aim of this study is to identify drug target candidates of A. pleuropneumoniae from essential proteins in PPI network. The homologous protein mapping method (HPM) was utilized to construct A. pleuropneumoniae PPI network. Afterwards, the subnetwork centered with H-NS was selected to verify the PPI network using bacterial two-hybrid assays. Drug target candidates were identified from the hub proteins by analyzing the topology of the network using interaction degree and homologous comparison with the pig proteome. An A. pleuropneumoniae PPI network containing 2737 non-redundant interaction pairs among 533 proteins was constructed. These proteins were distributed in 21 COG functional categories and 28 KEGG metabolic pathways. The A. pleuropneumoniae PPI network was scale free and the similar topological tendencies were found when compared with other bacteria PPI network. Furthermore, 56.3% of the H-NS subnetwork interactions were validated. 57 highly connected proteins (hub proteins) were identified from the A. pleuropneumoniae PPI network. Finally, 9 potential drug targets were identified from the hub proteins, with no homologs in swine. This study provides drug target candidates, which are promising for further investigations to explore lead compounds against A. pleuropneumoniae.
Isoforms of wild type proteins often appear as low molecular weight bands on SDS-PAGE.
Zhang, Ju,Lou, Xiaomin,Shen, Haihong,Zellmer, Lucas,Sun, Yuan,Liu, Siqi,Xu, Ningzhi,Liao, D Joshua Wiley 2014 Biotechnology Journal Vol.9 No.8
<P>Immunoblotting, after polyacrylamide gel electrophoresis with sodium dodecyl sulfate (SDS-PAGE), is a technique commonly used to detect specific proteins. SDS-PAGE often results in the visualization of protein band(s) in addition to the one expected based on the theoretical molecular mass (TMM) of the protein of interest. To determine the likelihood of additional band(s) being nonspecific, we used liquid chromatography - mass spectrometry to identify proteins that were extracted from bands with the apparent molecular mass (MM) of 40 and 26 kD, originating from protein extracts derived from non-malignant HEK293 and cancerous MDA-MB231 (MB231) cells separated using SDS-PAGE. In total, approximately 57% and 21% of the MS/MS spectra were annotated as peptides in the two cell samples, respectively. Moreover, approximately 24% and 36.2% of the identified proteins from HEK293 and MB231 cells matched their TMMs. Of the identified proteins, 8% from HEK293 and 26% from MB231 had apparent MMs that were larger than predicted, and 67% from HEK293 and 37% from MB231 exhibited smaller MM values than predicted. These revelations suggest that interpretation of the positive bands of immunoblots should be conducted with caution. This study also shows that protein identification performed by mass spectrometry on bands excised from SDS-PAGE gels could make valuable contributions to the identification of cancer biomarkers, and to cancer-therapy studies.</P>
Siqi Huang,Congcong Chen,Zhibo Zhao,Lingyi Jia,Yong Zhang 한국공업화학회 2023 Journal of Industrial and Engineering Chemistry Vol.118 No.-
In this work, hydroxyapatite (HAP) was in situ synthesized on coal fly ash (CFA) to prepare hydroxyapatitemodified coal fly ash composite aerogel (HCFAA). The maximum removal efficiency and capabilityof uranium(VI) on HCFAA were 97.6 % and 205.7 mg g1, respectively, which was much higher than thoseof CFA (83.6 % and 59.1 mg g1) (pH = 3.0, m/V = 1.0 g/L and T = 298 K). The desorption efficiency of uranium(VI) by HCFAA was even more than 80 % after five cycles, demonstrating that the introduction ofHAP had improved uranium(VI) removal performances. Pseudo-second-order and Langmuir models werefitted better with the experimental data, indicating the uranium(VI) removal process was a homogeneousmonolayer chemisorption. Meanwhile, the uranium(VI) removal efficiency for HCFAA in actual wastewaterwas higher to 80.6 % and uranium(VI) could even be completely separated from actual wastewaterby HCFAA during dynamic adsorption (m = 50 mg, V = 400 mL, C0 = 10 mg/L), further illustrating that theintroduction of HAP was an available method to modify CFA. Characterizations results demonstrated thaturanium(VI) was successfully immobilized on HCFAA through ion exchange, dissolution-precipitationand surface complexation. In conclusion, HCFAA was a prospective adsorbent for uranium(VI) separationin practical application.
Hao Zhang,Jing Sun,Xianting Liu,Chuan Hong,Yuanbo Zhu,Aiping Liu,Siqi Li,Huiyuan Guo,Fazheng Ren 한국미생물학회 2013 The journal of microbiology Vol.51 No.6
Lactobacillus paracasei subsp. paracasei LC01 (LC01) can tolerate intestinal stresses and has antioxidant activity. To evaluate the effect of the bacterium on human intestinal microflora,a randomized, double-blind, placebo-controlled human trial was carried out. Fifty-two healthy adult volunteers were randomized equally to two groups. One group consumed 12% (wt/vol) skimmed milk supplemented with 1010 CFU of LC01 each day for the 4-week treatment period,and then consumed placebo in the next treatment period,separated by a 2-week washout. The other group followed the reverse order. Group-specific real-time PCR and biochemical analyses was used to determine the intestinal bacterial composition of fecal samples collected at the end of every period, and the concentration of short-chain fatty acids and ammonia. A significant inhibition in fecal Escherichia coli and increase in Lactobacillus, Bifidobacterium, and Roseburia intestinalis were observed after consumption of LC01. Acetic acid and butyric acid were significantly higher in the probiotic stage and fecal ammonia was significantly lower. The results indicated a modulation effect of LC01 on the intestinal microflora of young adults, suggesting a beneficial effect on bowel health. LC01 may have potential value as a probiotic.
Xuyang Zhang,Hangjia Shen,Babak Rezaei Moghadam,Siqi Liu,Yuejin Zhu,Jiacheng Wang,Minghui Yang 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2019 NANO Vol.14 No.7
Practical application of oxygen evolution reaction (OER) in energy conversion devices, such as water splitting and metal-air batteries, relies on the development of highly efficient and stable electrocatalysts and is still challengeable. Here, Nickel modulated ternary Iron tungsten nitride (Ni-FeWN2) nanosheets are synthesized through a facile hydrothermal growth method and subsequently nitriding process for boosting OER. A potential of 1.57V at the current density of 10 mA/cm2 and Tafel slope (40 mV/dec) are achieved on Ni-FeWN2, much lower than that on Ni-free FeWN2 and commercial IrO2. More importantly, Ni-FeWN2 nanosheets display high stability and electrocatalytic activity toward driving oxygen evolution efficiently in alkaline media.