VSV-G Viral Envelope Glycoprotein Prepared from Pichia pastoris Enhances Transfection of DNA into Animal Cells

( Xin Liu ),( Ying Dong ),( Jingquan Wang ),( Long Li ),( Zhenmin-zhong ),( Yun-pan Li ),( Shao-jun Chen ),( Yu-cai Fu ),( Wen-can Xu ),( Chi-ju Wei ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.6

Vesicular stomatitis virus G glycoprotein (VSV-G) has been widely used for pseudotyping retroviral, lentiviral, and artificial viral vectors. The objective of this study was to establish a potential approach for large-scale production of VSV-G. To this end, VSV-G was cloned with an N-terminal His-tag into Pichia pastoris expression vector pPIC3.5K. Three clones (Mut^s) containing the VSV-G expression cassette were identified by PCR. All clones proliferated normally in expansion medium, whereas the proliferation was reduced significantly under induction conditions. VSV-G protein was detected in cell lysates by western blot analysis, and the highest expression level was observed at 96 h post induction. VSV-G could also be obtained from the condition medium of yeast protoplasts. Furthermore, VSV-G could be incorporated into Ad293 cells and was able to induce cell fusion, leading to the transfer of cytoplasmic protein. Finally, VSV-G-mediated DNA transfection was assayed by flow cytometry and luciferase measurement. Incubation of VSV-G lysate with the pGL3-control DNA complex increased the luciferase activity in Ad293 and HeLa cells by about 3-fold. Likewise, incubation of VSV-G lysate with the pCMV-DsRed DNA complex improved the transfection efficiency into Ad293 by 10% and into HeLa cells by about 1-fold. In conclusion, these results demonstrate that VSV-G could be produced from P. pastoris with biofunctionalities, demonstrating that large-scale production of the viral glycoprotein is feasible.

Serum Insulin-Like Growth Factor Binding Protein 7 as a Potential Biomarker in the Diagnosis and Prognosis of Esophagogastric Junction Adenocarcinoma

Can-Tong Liu,Yi-Wei Xu,Hong Guo,Chao-Qun Hong,Xin-Yi Huang,Yu-Hao Luo,Shi-Han Yang,Ling-Yu Chu,En- Min Li,Yu-Hui Peng 거트앤리버 소화기연관학회협의회 2020 Gut and Liver Vol.14 No.6

Background/Aims: Esophagogastric junction adenocarcinoma (EJA) is a malignant tumor associated with high morbidity and has attracted increasing attention due to a rising incidence and low survival rate. Pathological biopsy is the gold standard for diagnosis, but noninvasive and effective tests are lacking, resulting in diagnoses at advanced stages. This study explored the diagnostic value of insulin-like growth factor binding protein 7 (IGFBP7) in EJA. Methods: A total of 120 EJA patients and 88 normal controls were recruited, and their serum levels of IGFBP7 were measured by enzymelinked immunosorbent assay. Receiver operating characteristic (ROC) curve analysis was used to assess the diagnostic value, and Pearson chi-square analysis was used to evaluate the correlation between IGFBP7 and clinical parameters. Kaplan- Meier survival analysis was carried out to assess the effect of IGFBP7 on overall survival (OS). Results: The levels of IGFBP7 were higher in both early- and late-stage EJA patients than in normal controls (p<0.001). The area under the ROC curve for EJA patients was 0.794 (95% confidence interval [CI], 0.733 to 0.854), with a cutoff value of 2.716 ng/mL, a sensitivity of 63.3% (95% CI, 54.0% to 71.8%) and a specificity of 90.9% (95% CI, 82.4% to 95.7%). For the diagnosis of early-stage EJA, the same cutoff value and specificity were obtained, but the sensitivity of IGFBP7 was 54.3% (95% CI, 36.9% to 70.8%). Patients with low IGFBP7 protein expression had lower OS than those with high expression (p=0.034). The multivariate analysis showed that IGFBP7 is an independent prognostic factor for EJA (p=0.011). Conclusions: Serum IGFBP7 acts as a potential diagnostic and prognostic marker for EJA.

<i>Arcticibacter pallidicorallinus</i> sp. nov. isolated from glacier ice

Liu, Qing,Kim, Song-gun,Liu, Hong-can,Xin, Yu-hua,Zhou, Yu-guang International Union of Microbiological Societies 2014 International journal of systematic and evolutiona Vol.64 No.7

<P>A Gram-stain-negative, rod-shaped bacterium (strain Hh36<SUP>T</SUP>) was isolated from the No. 1 glacier in Xinjiang, north-west China. Colonies of strain Hh36<SUP>T</SUP> were pink, convex and round on PYG medium plates. Strain Hh36<SUP>T</SUP> was able to grow at 4–30 °C and pH 6.0–8.0. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Hh36<SUP>T</SUP> was related to members of the genus <I>Arcticibacter</I>. The major cellular fatty acids of the novel strain were iso-C<SUB>15 : 0</SUB>, summed feature 3 (C<SUB>16 : 1</SUB>ω6<I>c</I> and/or C<SUB>16 : 1</SUB>ω7<I>c</I>) and iso-C<SUB>17 : 0</SUB> 3-OH. The G+C content of the genomic DNA was 44.0 mol%. On the basis of phenotypic characteristics and phylogenetic analysis, strain Hh36<SUP>T</SUP> is considered to represent a novel species of the genus <I>Arcticibacter</I>, for which the name <I>Arcticibacter</I> <I>pallidicorallinus</I> sp. nov. is proposed. The type strain is Hh36<SUP>T</SUP> ( = CGMCC 1.9313<SUP>T</SUP> = KCTC 32542<SUP>T</SUP>).</P>

Relation between Ki-67, ER, PR, Her2/neu, p21, EGFR, and TOP II-α Expression in Invasive Ductal Breast Cancer Patients and Correlations with Prognosis

Yan, Jian,Liu, Xiao-Long,Han, Lu-Zhe,Xiao, Gang,Li, Ning-Lei,Deng, Yi-Nan,Yin, Liang-Chun,Ling, Li-Juan,Yu, Xiao-Yuan,Tan, Can-Liang,Huang, Xiao-Ping,Liu, Li-Xin Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.2

The aim of the present study was to investigate the expression of the transcription factor Ki-67, ER, PR, Her2/neu, p21, EGFR, and TOP II-${\alpha}$ in the tumor tissue of patients with invasive ductal carcinoma(IDC); in addition, we examined correlations between these markers. Two hundred and sixteen IDC patients, who were not previously been treated with chemo- or radiotherapy, were included in the study. All tumors were grade I-III. Expression of molecular markers was determined by immunohistochemical analysis on paraffin-embedded tissue sections. Follow-up data were collected for 3 months to 10 years and analyzed for tumor recurrence, survival time, and prognostic risk factors. We determined Ki-67 expression correlates with the expression of ER, PR, HER-2, EGFR, and TOP-${\alpha}$, as well as lymph node involvement, high tumor grade, lymphovascular invasion, high tumor stage, and high TNM stage in IDC. Positive Ki-67 expression was a risk factor for rapid tumor recurrence and may help tumor progression, leading to poor prognosis in IDC. Ki-67 was directly correlated with EGFR, TOP II-${\alpha}$, lymph node involvement, high tumor grade, lymphovascular invasion, high tumor stage, and high TNM stage in the hormone receptor subtypes of breast cancer. In triple negative breast cancer, Ki-67 correlated with TOP II-${\alpha}$. Expression of Ki-67 correlated with that of ER, PR, HER-2, EGFR, TOP II-${\alpha}$, and p21. In addition, the biomarker Ki-67 has a role as a prognostic factor and indicates a poor prognosis in IDC.

<i>Glaciihabitans tibetensis</i> gen. nov., sp. nov., a psychrotolerant bacterium of the family <i>Microbacteriaceae</i>, isolated from glacier ice water

Li, Ai-Hua,Liu, Hong-Can,Xin, Yu-Hua,Kim, Song-Gun,Zhou, Yu-Guang International Union of Microbiological Societies 2014 International journal of systematic and evolutiona Vol.64 No.2

<P>A Gram-stain-positive, aerobic, non-spore-forming, short-rod-shaped bacterium, designated strain MP203<SUP>T</SUP>, was isolated from ice water of Midui Glacier in Tibet Autonomous Region, China. The strain was psychrotolerant, growing at 0–25 °C. 16S rRNA gene sequence analysis showed that strain MP203<SUP>T</SUP> was most similar to <I>Frigoribacterium faeni</I> NBRC 103066<SUP>T</SUP>, <I>Compostimonas suwonensis</I> KACC 13354<SUP>T</SUP>, <I>Frigoribacterium mesophilum</I> KCTC 19311<SUP>T</SUP>, <I>Marisediminicola antarctica</I> CCTCC AB 209077<SUP>T</SUP> and <I>Alpinimonas psychrophila</I> JCM 18951<SUP>T</SUP>, with similarities of 97.4, 97.2, 97.2, 97.1 and 97.1 %, respectively. The maximum-likelihood phylogenetic tree indicated that strain MP203<SUP>T</SUP> clustered with nine genera of the family <I>Microbacteriaceae</I>, namely <I>Frigoribacterium</I>, <I>Compostimonas</I>, <I>Marisediminicola</I>, <I>Alpinimonas</I>, <I>Frondihabitans</I>, <I>Clavibacter</I>, <I>Subtercola</I>, <I>Klugiella</I> and <I>Agreia</I><I>.</I> However, bootstrap analysis showed that there was no significance in the branching pattern of the linage comprising strain MP203<SUP>T</SUP> and any existing generic lineage of the family <I>Microbacteriaceae</I>. DNA–DNA hybridization results indicated levels of relatedness between strain MP203<SUP>T</SUP> and <I>Marisediminicola antarctica</I> CCTCC AB 209077<SUP>T</SUP>, <I>Frigoribacterium faeni</I> NBRC 103066<SUP>T</SUP>, <I>Frigoribacterium mesophilum</I> KCTC 19311<SUP>T</SUP>, <I>Compostimonas suwonensis</I> KACC 13354<SUP>T</SUP> and <I>Alpinimonas psychrophila</I> JCM 18951<SUP>T</SUP> were 25.8±7.3, 29.6±7.6, 19.7±6.7, 16.0±4.2 and 12.4±5.1 % (mean±<SMALL>sd</SMALL>), respectively. The G+C content of the genomic DNA was 64.1 mol%. Analysis of the cell-wall peptidoglycan revealed that the peptidoglycan structure of strain MP203<SUP>T</SUP> was B10 type with Gly[<SMALL>l</SMALL>-Hse]–<SMALL>d</SMALL>-Glu–<SMALL>d</SMALL>-DAB, containing 2, 4-diaminobutyric acid (DAB) as a diagnostic amino acid. The cell-wall sugars were rhamnose, ribose, mannose and glucose. The major fatty acids were anteiso-C<SUB>15 : 0</SUB>, iso-C<SUB>16 : 0</SUB> and anteiso A-C<SUB>15 : 1</SUB>. An unusual compound identified as anteiso-C<SUB>15 : 0</SUB>-DMA (1, 1-dimethoxy-anteiso-pentadecane) was also present in strain MP203<SUP>T</SUP>. The predominant menaquinone was MK-10. Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), one unknown glycolipid and four unknown lipids were detected in the polar lipid extracts. As strain MP203<SUP>T</SUP> was distinguishable from phylogenetically related genera in the family <I>Microbacteriaceae</I> in terms of its physiological and chemotaxonomic characteristics and phylogenetic position, it was considered to represent a novel species of a new genus. Thus, the name <I>Glaciihabitans tibetensis</I> gen. nov., sp. nov. is proposed. The type strain of <I>Glaciihabitans tibetensis</I> is MP203<SUP>T</SUP> ( = CGMCC 1.12484<SUP>T</SUP> = KCTC 29148<SUP>T</SUP>).</P>

Keratin 17 identified by proteomic analysis may be involved in tumor angiogenesis

( Yong Xu ),( Su Zhen Zhang ),( Can Hua Huang ),( Xin Yu Liu ),( Zhen Hua Zhong ),( Wen Li Hou ),( Zi Fen Su ),( Yu Quan Wei ) 생화학분자생물학회 2009 BMB Reports Vol.42 No.6

H-형태 양친매성 펜타블록 공중합체의 화학효소적 합성과 자기회합거동 평가

Peng Chen,Ya Peng Li,Can Jin Li,Xin Lei Meng,Bao Zhang,Ming Zhu,Yan-jing Liu,Jing Yuan Wang 한국고분자학회 2013 폴리머 Vol.37 No.3

H-shaped amphiphilic pentablock copolymers (PSt)2-b-PCL-b-PEO-b-PCL-b-(PSt)2 was synthesized via chemoenzymatic method by combining enzyme-catalyzed ring-opening polymerization (eROP) of ε-caprolactone (ε-CL) and atom transfer radical polymerization (ATRP) of styrene. By this process, we obtained copolymers with controlled molecular weight and low polydispersity. The structure and composition of the obtained copolymers were characterized by nuclear magnetic resonance (NMR), gel permeation chromatography (GPC) and infrared spectroscopy analysis (IR). The crystallization behavior of the copolymers was analyzed by differential scanning calorimetry (DSC) and X-ray diffraction (XRD). The crystallization behavior of the H-shaped block copolymers demonstrated a PCL dominate crystallization. The self-assembly behavior of the copolymers was investigated in aqueous media. The hydrodynamic diameters of the copolymer micelles in aqueous solution were measured by dynamic light scattering (DLS). The morphology of the copolymer micelles was observed by atomic force microscopy (AFM) and transmission electron microscopy (TEM). The hydrodynamic diameters of spherical micelles declined gradually with the increase of the hydrophobic chain lengths of the copolymers. The critical micelle concentration (CMC) values were determined from fluorescence emission, and it was found that the CMCs decreased with an increase of PSt hydrophobic block lengths.

Valsartan regulates TGF-b/Smads and TGF-b/p38 pathways through lncRNA CHRF to improve doxorubicin-induced heart failure

Lei Chen,Kui-Po Yan,Xin-Can Liu,Wei Wang,Chao Li,Ming Li,Chun-Guang Qiu 대한약학회 2018 Archives of Pharmacal Research Vol.41 No.1

This study investigated the interaction amongvalsartan (VAL), TGF-b pathways, and long non-codingRNA (lncRNA) cardiac hypertrophy-related factor (CHRF)in doxorubicin (DOX)-induced heart failure (HF), andexplored their roles in DOX-induced HF progression. HFmice models in vivo were constructed by DOX induction. The expression of CHRF and TGF-b1 in hearts wasdetected, along with cardiac function, caspase-3 activity,and cell apoptosis. Primary myocardial cells were pretreatedwith VAL, followed by DOX induction in vitro forfunctional studies, including the detection of cell apoptosiswith terminal deoxynucleotidyl transferase dUTP nick-endlabeling and the expression of proteins associated withTGF-b1 pathways. HF models were established in vivo andin vitro. Expression of CHRF and TGF-b1 was up-regulated,and cell apoptosis and caspase-3 activity wereincreased in the hearts and cells of the HF models. VALsupplementation alleviated the cardiac dysfunction andinjury in the HF process. Moreover, overexpressed CHRFup-regulated TGF-b1, promoted myocardial cell apoptosis,and reversed VAL’s cardiac protective effect, while interferenceof CHRF (si-CHRF) did the opposite. Down-regulationof CHRF reversed the increased expression of TGFb1and the downstream proteins induced by pcDNA-TGFb1in HL-1 cells, while overexpression of CHRF reversedthe VAL’s cardiac protective effect in vivo. In conclusion,VAL regulates TGF-b pathways through lncRNA CHRF toimprove DOX-induced HF.