Protoplast Preparation and Polyethylene Glycol (PEG)-mediated Transformation of Candida glycerinogenes

Cheng Zhang,Hong Zong,Bin Zhuge,Xinyao Lu,Huiying Fang,Jiali Zhu,Jian Zhuge 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

The regeneration of Candida glycerinogenes protoplasts is a major step following genetic manipulations such as fusion and DNA-mediated transformation. An investigation of protoplast formation and cytological examination was used to gain further insight into the loss of protoplast viability in osmotically stabilized support media. Protoplasts with the highest regeneration frequency (98.6% protoplasts/mL) were isolated, using lysozyme dissolved in 1M sorbitol osmoticum. The commercial enzyme preparations, osmotic stabilisers, and growth phase were effective in raising the protoplast yield. Sodium chloride was effective for protoplast preparation; however, sugars and sugar alcohols were better for protoplast regeneration. Sorbitol at a concentration of 1 M was used in regeneration agar for further studies. Regeneration of colonies from protoplasts was maximal (11 ~ 15%) when protoplasts were incorporated in cooled agar containing 0.5% glucose, supplemented with 1M sorbitol as osmotic stabilizer. C. glycerinogenes strain was highly sensitive to zeocin, so transformation of protoplasts and PEG-mediated was achieved with an improved transformation system, using plasmid pURGAP-gfp containing zeocin gene driven by a PCgGAP promoter from C. glycerinogenes to express gfp gene and be transformed into the 5.8S rDNA site of C. glycerinogenes in order to test the system for studying the yeast osmoregulation. We developed an efficient method for transformation of C. glycerinogenes, and parameters involved in transformation efficiency were optimized. Expressions of gfp at different levels were conducted under osmotic stress containing NaCl, KCl, sorbitol or glycerol for the recombinant strains. These improved procedures for protoplast isolation, regeneration and transformation proved to be useful applications in genetic studies for other Candida species and industrial yeast.

Histone Deacetylase Inhibitor Trichostatin A Enhances Antitumor Effects of Docetaxel or Erlotinib in A549 Cell Line

Zhang, Qun-Cheng,Jiang, Shu-Juan,Zhang, Song,Ma, Xiao-Bin Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.7

Background and Objective: Histone deacetylase (HDAC) inhibitors represent a promising class of potential anticancer agents for treatment of human malignancies. In this study, we investigated the effect of trichostatin A (TSA), one such HDAC inhibitor, in combination with docetaxel (TXT), a cytotoxic chemotherapy agent or erlotinib, a novel molecular target therapy drug, on lung cancer A549 cells. Methods: A549 cells were treated with TXT, erlotinib alone or in combination with TSA, respectively. Cell viability, apoptosis, and cell cycle distribution were evaluated using MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assay, Hochst33258 staining and flow cytometry. Moreover, immunofluorescent staining and Western blot analysis were employed to examine alterations of ${\alpha}$-tubulin, heat shock protein 90 (hsp90), epidermal growth factor receptor (EGFR), and caspase-3 in response to the different exogenous stimuli. Results: Compared with single-agent treatment, co-treatment of A549 cells with TSA/TXT or TSA/erlotinib synergistically inhibited cell proliferation, induced apoptosis, and caused cell cycle delay at the $G_2/M$ transition. Treatment with TSA/TXT or TSA/erlotinib led to a significant increase of cleaved caspase-3 expression, also resulting in elevated acetylation of ${\alpha}$-tubulin or hsp90 and decreased expression of EGFR, which was negatively associated with the level of acetylated hsp90. Conclusions: Synergistic anti-tumor effects are observed between TXT or erlotinib and TSA on lung cancer cells. Such combinations may provide a more effective strategy for treating human lung cancer.

Research on the Impact of Agricultural Mechanization Service on Wheat Planting Cost: A Case Study of Henan Province

Zhang Cheng 한국정보처리학회 2021 Journal of information processing systems Vol.17 No.6

Given the different effects of agricultural mechanization on various stages of wheat planting in Henan, this article selects 78 observation samples from Henan, a major wheat-growing province. It uses different research methods (multiple linear regression, social network analysis model, multi-layer sensory nerves network) to conduct a comparative study, and the calculation results of the model show that the experimental results have a strong convergence and consistency. Agricultural mechanization services have significant effects on the three stages of wheat planting: harvesting, plowing and sowing. A higher degree of mechanized service in several stages can reduce the cost of growing wheat on family farms.

Bicluster and Pathway Enrichment Analysis of HCV-induced Cirrhosis and Hepatocellular Carcinoma

Cheng, Peng,Cheng, You,Su, Mei X.,Li, Dong,Zhao, Guo Z.,Gao, Hui,Li, Yan,Zhu, Jie Y.,Li, Hua,Zhang, Tao Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Background: Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the most common form of liver cancer. However, while it is associated frequently with hepatitis C virus (HCV) there is only an elementary understanding of its molecular pathogenesis. Methods: To gain insight into the molecular mechanisms of HCV-induced hepatocarcinogenesis, we performed microarray analysis on 75 surgical liver samples from 48 HCV-infected patients. Results: There were 395 differentially expressed geness between cirrhotic samples and HCC samples. Of these, 125 genes were up-regulated and 270 genes were down-regulated. We performed pathway enrichment analysis and screened as described previously. Conclusions: The differentially expressed genes might be involved in hepatocarcinogenesis through upregulating the pathways of ECM-receptor interaction, focal adhesion, cell adhesion molecules and other cancer-related pathways, and downregulating the pathways of "complement and coagulation cascades". We hope our results could aid in seeking of therapeutic targets for HCV-induced hepatocellular carcinoma.

Effects of different heating methods on the contents of nucleotides and related compounds in minced Pacific white shrimp and Antarctic krill

Zhang, Ruijuan,Qiu, Weiqiang,Zhang, Maosai,Ho Row, Kyung,Cheng, Yudong,Jin, Yinzhe Elsevier 2018 FOOD SCIENCE AND TECHNOLOGY -ZURICH- Vol.87 No.-

<P>The effect of heating methods (microwave and water bath, and combined microwave and water bath heating) on 6 nucleotides and related compounds (NRCs) in minced Pacific white shrimp and Antarctic krill was investigated at 25, 40, 55, 65, 75, 85, and 95 degrees C. In Pacific white shrimp, the main changes of NRCs were in adenosine monophosphate (AMP) and inosine monophosphate (IMP). High temperature (85 and 95 degrees C) can inhibit the ATP degradation by water bath heating. In Antarctic krill, the main change of NRCs was in inosine (HxR) that decreased in microwave heating treatment at 95 degrees C. The different methods were compared in terms of IMP content and flavor/bitter ratio, which are related to flavor of aquatic products. The best heating method was pre-heating at 75 degrees C in water bath followed by microwave heating at 95 degrees C (IMP content and relatively flavor/bitter ratio were 4.34 mu mol/g and 3.00, respectively) for pacific white shrimp and pre-heating at 65 degrees C in microwave followed by water bath heating at 95 degrees C (IMP content and relatively flavor/bitter ratio were 0.18 mu mol/g and 0.06, respectively) for Antarctic krill, respectively. Chemical analysis, microbiological analysis and sensory evaluation showed that the K value is not suitable for assessing the freshness of Antarctic krill. (C) 2017 Elsevier Ltd. All rights reserved.</P>

Determination of Survival of Gastric Cancer Patients With Distant Lymph Node Metastasis Using Prealbumin Level and Prothrombin Time: Contour Plots Based on Random Survival Forest Algorithm on High-Dimensionality Clinical and Laboratory Datasets

Zhang, Cheng,Xie, Minmin,Zhang, Yi,Zhang, Xiaopeng,Feng, Chong,Wu, Zhijun,Feng, Ying,Yang, Yahui,Xu, Hui,Ma, Tai The Korean Gastric Cancer Association 2022 Journal of gastric cancer Vol.22 No.2

Purpose: This study aimed to identify prognostic factors for patients with distant lymph node-involved gastric cancer (GC) using a machine learning algorithm, a method that offers considerable advantages and new prospects for high-dimensional biomedical data exploration. Materials and Methods: This study employed 79 features of clinical pathology, laboratory tests, and therapeutic details from 289 GC patients whose distant lymphadenopathy was presented as the first episode of recurrence or metastasis. Outcomes were measured as any-cause death events and survival months after distant lymph node metastasis. A prediction model was built based on possible outcome predictors using a random survival forest algorithm and confirmed by 5×5 nested cross-validation. The effects of single variables were interpreted using partial dependence plots. A contour plot was used to visually represent survival prediction based on 2 predictive features. Results: The median survival time of patients with GC with distant nodal metastasis was 9.2 months. The optimal model incorporated the prealbumin level and the prothrombin time (PT), and yielded a prediction error of 0.353. The inclusion of other variables resulted in poorer model performance. Patients with higher serum prealbumin levels or shorter PTs had a significantly better prognosis. The predicted one-year survival rate was stratified and illustrated as a contour plot based on the combined effect the prealbumin level and the PT. Conclusions: Machine learning is useful for identifying the important determinants of cancer survival using high-dimensional datasets. The prealbumin level and the PT on distant lymph node metastasis are the 2 most crucial factors in predicting the subsequent survival time of advanced GC.

Distinctive Roles of Wnt Signaling in Chondrogenic Differentiation of BMSCs under Coupling of Pressure and Platelet-Rich Fibrin

Cheng Baixiang,Feng Fan,Shi Fan,Huang Jinmei,Zhang Songbai,Quan Yue,Tu Teng,Liu Yanli,Wang Junjun,Zhao Ying,Zhang Min 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.4

BACKGROUND: Although newly formed constructs of feasible pressure-preadjusted bone marrow mesenchymal stem cells (BMSCs) and platelet-rich fibrin (PRF) showed biomechanical flexibility and superior capacity for cartilage regeneration, it is still not very clear how BMSCs and seed cells feel mechanical stimuli and convert them into biological signals, and the difference in signal transduction underlying mechanical and chemical cues is also unclear. METHODS: To determine whether mechanical stimulation (hydrostatic pressure) and chemical cues (platelet-rich fibrin, PRF) activate canonical or noncanonical Wnt signaling in BMSCs, BMSCs cocultured with PRF were subjected to hydrostatic pressure loading, and the activation of the Wnt signaling molecules and expression of cartilage-associated proteins and genes were determined by western blotting and polymerase chain reaction (PCR). Inhibitors of canonical or noncanonical Wnt signaling, XVX-939 or L690,330, were adopted to investigate the role of Wnt signaling molecules in mechanically promoted chondrogenic differentiation of BMSCs. RESULTS: Hydrostatic pressure of 120 kPa activated both Wnt/b-catenin signaling and Wnt/Ca2? signaling, with the the maximum promotion effect at 60 min. PRF exerted no synergistic effect on Wnt/b-catenin signaling activation. However, the growth factors released by PRF might reverse the promotion effects of pressure on Wnt/Ca2? signaling. Real-time PCR and Western blotting results showed that pressure could activate the expression of Col-II, Sox9, and aggrecan in BMSCs cocultured with PRF. Blocking experiment found a positive role of Wnt/b-catenin signaling, and a negative role of Wnt/ Ca2? signaling in chondrogenic differentiation of the BMSCs. Mutual inhibition exists between canonical and noncanonical Wnt signaling in BMSCs under pressure. CONCLUSION: Wnt signaling participates in the pressure-promoted chondrogenesis of the BMSCs co-cultured with PRF, with canonical and noncanonical pathways playing distinct roles during the process.

Transjugular intrahepatic portosystemic shunt assisted with trans-splenic access

Zhang Cheng,Feng Wang,Jianfei Liu 대한간학회 2019 Clinical and Molecular Hepatology(대한간학회지) Vol.25 No.4

Regulating the electronic structures of mixed B-site pyrochlore to enhance the turnover frequency in water oxidation

Zhang Cheng,Wang Fangfang,Xiong Beichen,Yang Hong 나노기술연구협의회 2022 Nano Convergence Vol.9 No.22

This paper describes the development of mixed B-site pyrochlore Y2MnRuO7 electrocatalyst for oxygen evolution reaction (OER) in acidic media, a challenge for the development of low-temperature electrolyzer for green hydrogen production. Recently, several theories have been developed to understand the reaction mechanism for OER, though there is an uncertainty in most of the cases, due to the complex surface structures. Several key factors such as lattice oxygen, defect, electronic structure, oxidation state, hydroxyl group and conductivity were identified and shown to be important to the OER activity. The contribution of each factor to the performance however is often not well understood, limiting their impact in guiding the design of OER electrocatalysts. In this work, we showed mixed B-site pyrochlore Y2MnRuO7 catalyst exhibits 14 times higher turnover frequency (TOF) than RuO2 while maintaining a low overpotential of ~ 300 mV for the entire testing period of 24 h in acidic electrolyte. X-ray photoelectron spectroscopy (XPS) analysis reveals that this B-site mixed pyrochlore Y2MnRuO7 has a higher oxidation state of Ru than those of Y2Ru2O7, which could be crucial for improving OER performance as the broadened and lowered Ru 4d band resulted from the B-site substitution by Mn is beneficial to the OER kinetics.

Risk Assessment and Decision-Making of a Listed Enterprise’s L/C Settlement Based on Fuzzy Probability and Bayesian Game Theory

Zhang Cheng,Nanni Huang 한국정보처리학회 2020 Journal of information processing systems Vol.16 No.2

Letter of Credit (L/C) is currently a very popular international settlement method frequently used ininternational trade processes amongst countries around the globe. Compared with other international settlementmethods, however, L/C has some obvious shortcomings. Firstly, it is not easy to use due to the sophisticatedprocesses its usage involves. Secondly, it is sometimes accompanied by a few risks and some uncertainty. Thus,highly efficient methods need to be used to assess and control these risks. To begin with, FAHP and KMVmethods are used to resolve the problem of incomplete information associated with L/C and then, on this basis,Bayesian game theory is used in order to make more scientific and reasonable decisions with respect tointernational trade.