Inhibition of the NEDD8 Conjugation Pathway by shRNA to UBA3, the Subunit of the NEDD8-Activating Enzyme, Suppresses the Growth of Melanoma Cells

Cheng, Fang,Chen, Hao,Zhang, Lei,Ruo-Hong, Li,Liu, Yi,Sun, Jian-Fang Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.1

Neural precursor cell-expressed developmentally down-regulated 8 (NEDD8), a ubiquitin-like protein, mainly functions through covalent ligation to cullin proteins. Conjugation of NEDD8 with cullins can promote ubiquitination, which plays a critical role in the degradation of many proteins. UBA3 is the subunit of NEDD8-activating enzyme which is one of the keys for NEDD8 linkage to cullin proteins. Previous research showed NEDD8 conjugation to be up-regulated in highly proliferative cell lines. In the present study, up-regulated NEDD8 conjugation was observed in melanoma cell lines by Western blot analysis. After down-regulation with a RNAi to UBA3, proliferation of M14 was suppressed in vitro and in vivo. In conclusion, up-regulated NEDD8 conjugation may be involved in the development of melanoma. Interference in this pathway might offera promising method for melanoma therapy.

A Swimming Hydroponic System for Leafy Vegetable Production (ZAAS-SwimHydro)

Zhi-hao Xu,Wei-shon Shou,Cheng-hao Zhang,Wen-qi Dong,Jue-li Lei,Jian Cheng,Sheng-jun Zhou,Yu-qian Zhu,Dang-li Dai,Li-ping Cheng,Jian Xu,Jin-chun Fang,Sang Gyu Lee,Myeong-Cheoul Cho 한국원예학회 2008 한국원예학회 기타간행물 Vol.- No.-

Genetic Organization of ascB-dapE Internalin Cluster Serves as a Potential Marker for Listeria monocytogenes Sublineages IIA, IIB, and IIC

( Jian Shun Chen ),( Chun Fang ),( Ning Yu Zhu ),( Yong Hui Lv ),( Chang Yong Cheng ),( Yi Jiang Bei ),( Tianlun Zheng ),( Wei Huan Fang ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.5

Listeria monocytogenes is an important foodborne pathogen that comprises four genetic lineages: I, II, III, and IV. Of these, lineage II is frequently recovered from foods and environments and responsible for the increasing incidence of human listeriosis. In this study, the phylogenetic structure of lineage II was determined through sequencing analysis of the ascB-dapE internalin cluster. Fifteen sequence types proposed by multilocus sequence typing based on nine housekeeping genes were grouped into three distinct sublineages, IIA, IIB, and IIC. Organization of the ascBdapE internalin cluster could serve as a molecular marker for these sublineages, with inlGHE, inlGC2DE, and inlC2DE for IIA, IIB, and IIC, respectively. These sublineages displayed specific genetic and phenotypic characteristics. IIA and IIC showed a higher frequency of recombination (ρ/θ). However, recombination events had greater effect (r/m) on IIB, leading to its high nucleotide diversity. Moreover, IIA and IIB harbored a wider range of internalin and stress-response genes, and possessed higher nisin tolerance, whereas IIC contained the largest portion of low-virulent strains owing to premature stop codons in inlA. The results of this study indicate that IIA, IIB, and IIC might occupy different ecological niches, and IIB might have a better adaptation to a broad range of environmental niches.

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.

Silencing of long noncoding RNA HOXA11-AS inhibits the Wnt signaling pathway via the upregulation of HOXA11 and thereby inhibits the proliferation, invasion, and self-renewal of hepatocellular carcinoma stem cells

Jun-Cheng Guo,Yi-Jun Yang,Jin-Fang Zheng,Jian-Quan Zhang,Min Guo,Xiang Yang,Xiang-Ling Jiang,Li Xiang,You Li,Huang Ping,Liu Zhuo 생화학분자생물학회 2019 Experimental and molecular medicine Vol.51 No.-

Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths, but its molecular mechanisms are not yet well characterized. Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis, including that of HCC. However, the role of homeobox A11 antisense (HOXA11-AS) in determining HCC stem cell characteristics remains to be explained; hence, this study aimed to investigate the effects of HOXA11-AS on HCC stem cell characteristics. Initially, the expression patterns of HOXA11-AS and HOXA11 in HCC tissues, cells, and stem cells were determined. HCC stem cells, successfully sorted from Hep3B and Huh7 cells, were transfected with short hairpin or overexpression plasmids for HOXA11-AS or HOXA11 overexpression and depletion, with an aim to study the influences of these mediators on the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo. Additionally, the potential relationship and the regulatory mechanisms that link HOXA11-AS, HOXA11, and the Wnt signaling pathway were explored through treatment with Dickkopf-1 (a Wnt signaling pathway inhibitor). HCC stem cells showed high expression of HOXA11-AS and low expression of HOXA11. Both HOXA11-AS silencing and HOXA11 overexpression suppressed the self-renewal, proliferation, migration, and tumorigenicity of HCC stem cells in vivo, as evidenced by the decreased expression of cancer stem cell surface markers (CD133 and CD44) and stemness-related transcription factors (Nanog, Sox2, and Oct4). Moreover, silencing HOXA11-AS inactivated the Wnt signaling pathway by decreasing the methylation level of the HOXA11 promoter, thereby inhibiting HCC stem cell characteristics. Collectively, this study suggested that HOXA11-AS silencing exerts an antitumor effect, suppressing HCC development via Wnt signaling pathway inactivation by decreasing the methylation level of the HOXA11 promoter.

Intracerebral Microdialysis Technique and its Application on Brain Pharmacokinetic- Pharmacodynamic Study

Yue-fang Pan,Jian Feng,Qiao-yuan Cheng,Fan-zhu Li 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.12

Intracerebral microdialysis (IC-MD) has been developed as a well-validated and powerful technique for decades. As a practical sampling tool, it can gain the continuous dialysates of endogenous and exogenous substances in extracellular fluid (ECF) of awake freely moving animals. Also, variform IC-MD probes (IC-MDPs) have grown more exquisite. The implantation of the IC-MDP in certain tissue of brain allows monitor drug distribution and measure drug and corresponding neurotransmitters levels in brain ECF after administration for brain pharmacokineticpharmacodynamic (B-PK-PD) study. So it is suitable for IC-MD to B-PK-PD study (IC-MD/BPK- PD). The performance of IC-MD/B-PK-PD can not only elevate the degree of precision and accuracy of experimental data, minimize the individual difference by reduced number of animals, but also give important information for the prediction and optimization of drug effective dose in preclinical study. In this review, we have discussed various IC-MD/B-PK-PD studies of analgesic, antiepileptic and antidepressant drug. The role of IC-MD/B-PK-PD in confirming and assessing the drug effect before clinic trials is highlighted.

Intracerebral Microdialysis Technique and its Application on Brain Pharmacokinetic-Pharmacodynamic Study

Pan, Yue-Fang,Feng, Jian,Cheng, Qiao-Yuan,Li, Fan-Zhu 대한약학회 2007 Archives of Pharmacal Research Vol.30 No.12

Intracerebral microdialysis (IC-MD) has been developed as a well-validated and powerful technique for decades. As a practical sampling tool, it can gain the continuous dialysates of endogenous and exogenous substances in extracellular fluid (ECF) of awake freely moving animals. Also, variform IC-MD probes (IC-MDPs) have grown more exquisite. The implantation of the IC-MDP in certain tissue of brain allows monitor drug distribution and measure drug and corresponding neurotransmitters levels in brain ECF a after administration for brain pharmacokinetic-pharmacodynamic (B-PK-PD) study. So it is suitable for IC-MD to B-PK-PD study (IC-MD/B-PK-PD). The performance of IC-MD/B-PK-PD can not only elevate the degree of precision and accuracy of experimental data, minimize the individual difference by reduced number of animals, but also give important information for the prediction and optimization of drug effective dose in preclinical study. In this review, we have discussed various IC-MD/B-PK-PD studies of analgesic, antiepileptic and antidepressant drug. The role of IC-MD/B-PK-PD in confirming and assessing the drug effect before clinic trials is highlighted.

Rapid preparation and characterization of chitosan nanoparticles for oligonucleotide

Mu-hua Cheng,Yao-xiong Huang,Han-jian Zhou,Zhi Liu,Jian-fang Li 한국물리학회 2010 Current Applied Physics Vol.10 No.3

Chitosan is regarded as one of the potential candidates as a gene carrier. However, the poor solubility of chitosan is the major limiting factor in its utilization as a gene carrier. The purpose of this study was to simplify the method of preparing the nanoparticles of chitosan linked with antisense oligonucleotide (asON). The main step was preparing the derivatives of chitosan phosphate (CSP) in order to easily dissolve in aqueous solution. The nanoparticles were formed using a simple mixed method for CSP and asON, and the nanoparticle’s forming condition was optimized so that the nanoparticle’s characterization could be examined. Results showed that it was simple to make the nanoparticles under the optimal condition of 2:1 M proportion of CSP and asON. The size of the nanoparticles was 102.6 ± 12.0 nm, its zeta potential was 1.45 ± 1.75, and the encapsulated ratio of the chitosan crosslinked the asON was 87.6 ± 3.5%. The infrared spectra and electron microscope displayed that chitosan may combine with the asON to form equirotal nanoparticles. In conclusion, it was simple and feasible to form chitosan nanoparticles for asON using the CSP, and the CSP can efficiently encapsulate asON.

Prognostic value of metabolic tumor volume and total lesion glycolysis from 18F-FDG PET/CT in lymph node metastases and risk stratification of endometrial carcinoma

Dou-dou Liu,Jian-fang Li,Xiaomao Li,Liangjun Xie,Luping Qin,Fangyu Peng,Mu-hua Cheng 대한부인종양학회 2019 Journal of Gynecologic Oncology Vol.30 No.6

Objective: To investigate the prognostic value of metabolic tumor volume (MTV) and totallesion glycolysis (TLG), measured by preoperative 18F-fluorodeoxyglucose positron emissiontomography/computed tomography (18F-FDG PET/CT), in risk stratification of patients withendometrial carcinoma (EC). Methods: The patients with pathological diagnosis of EC who underwent preoperative18F-FDG PET/CT imaging were retrospectively selected for analysis of the prognostic valuesof PET parameters in risk classification and lymph node metastases (LNMs). Receiveroperating-characteristic analysis was used to analyze the correlation of PET parameters cutoffvalues with deep myometrial invasion (MI), lymphovascular space involvement and LNM forprognostic values in risk stratification. Results: The sensitivity, specificity, positive predictive value, negative predictive value andaccuracy for detection of LNM are 83.3%, 99.7%, 90.9%, 99.5% and 99.2%, respectively. TheMTV and TLG of primary lesion of EC in the patients with LNM are notably higher than thosein patients without LNM, p<0.010. The MTV and TLG of the EC primary lesions in high-riskpatients are significantly higher than those in low-risk patients (p<0.010), but the maximumstandardized uptake value (SUVmax) is not. The MTV and TLG of primary lesions weresuperior to SUVmax for predicting of deep MI, LNM and high-risk of EC (p<0.005). Conclusion: MTV and TLG of primary lesions are more valuable in predicting riskstratification of EC patients. Preoperative 18F-FDG PET/CT imaging is useful in predictingthe LNM of EC and may help guide pelvic lymphadenectomy to avoid unnecessary pelviclymphadenectomy in EC patients with low-risk stratification.

miR-19a Promotes Cell Growth and Tumorigenesis through Targeting SOCS1 in Gastric Cancer

Qin, Shuang,Ai, Fang,Ji, Wei-Fang,Rao, Wang,Zhang, He-Cheng,Yao, Wen-Jian Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.2

Accumulating evidence has shown that microRNAs are involved in cancer development and progression. However, it remains unknown about the potential role of miR-19a in the pathogenesis of gastric cancer. Here, we report that suppressor of cytokine signaling 1 (SOCS1) is a novel target of miR-19a in gastric cancer cells and that miR-19a expression is inversely correlated with SOCS1 expression in gastric cancer cells and a subset of gastric cancer tissues. Ectopic expression of miR-19a dramatically promoted proliferation and tumorigenicity of gastric cancer cells both in vitro and in vivo. Moreover, we showed that silencing of SOCS1 promoted cell growth and colony formation resembling that of miR-19a overexpression, whereas re-introduction of SOCS1 (without the 3'-UTR) attenuated the pro-tumorigenic functions. Taken together, our findings suggest that the SOCS1 gene is a direct target of miR-19a, which functions as an oncogenic miRNA in gastric cancer by repressing the expression of tumor suppressor SOCS1.