Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

Shi, Qing-Qiang,Zuo, Guo-Wei,Feng, Zi-Qiang,Zhao, Lv-Cui,Luo, Lian,You, Zhi-Mei,Li, Dang-Yang,Xia, Jing,Li, Jing,Chen, Di-Long Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.18

Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

Cyclic Loading Tests on Composite Joints with Flush End Plate Connections

SHI Wen-long,LI Guo-qiang,YE Zhi-ming,R. Y. Xiao 한국강구조학회 2007 International Journal of Steel Structures Vol.7 No.2

The tests on composite joint specimens with flush end plate connections subjected to cyclic loads have been conducted inTongji University. One bare steel joint specimen has also been tested for comparison. Various instrumentations have been usedto measure: beam strains, column strains, rebar strains, connection deformations and deflections of specimens. The testobservations are reported in detail in this paper. The main results are as folows: (1) the moment-rotation relationships of thenability; (2) the composite joint specimens show large strength resistance and god ductility, and all the rotations of conectionare greater than 0.03rad as required by the FEMA-97; (3) similar failure modes have been identified from the observation ofthe composite specimens, which are found to be concentrated around the joint zone; and (4) the slipage between the concreteslab and steel beam is very small, which shows that betwen the concrete slab and steel beam the ful composition can be

Diversity of Endophytic Fungi from Different Verticillium-Wilt-Resistant Gossypium hirsutum and Evaluation of Antifungal Activity Against Verticillium dahliae In Vitro

( Zhi Fang Li ),( Ling Fei Wang ),( Zi Li Feng ),( Li Hong Zhao ),( Yong Qiang Shi ),( He Qin Zhu ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.9

Cotton plants were sampled and ranked according to their resistance to Verticillium wilt. In total, 642 endophytic fungi isolates representing 27 genera were recovered from Gossypium hirsutum root, stem, and leaf tissues, but were not uniformly distributed. More endophytic fungi appeared in the leaf (391) compared with the root (140) and stem (111) sections. However, no significant difference in the abundance of isolated endophytes was found among resistant cotton varieties. Alternaria exhibited the highest colonization frequency (7.9%), followed by Acremonium (6.6%) and Penicillium (4.8%). Unlike tolerant varieties, resistant and susceptible ones had similar endophytic fungal population compositions. In three Verticillium-wilt-resistant cotton varieties, fungal endophytes from the genus Alternaria were most frequently isolated, followed by Gibberella and Penicillium. The maximum concentration of dominant endophytic fungi was observed in leaf tissues (0.1797). The evenness of stem tissue endophytic communities (0.702) was comparatively more uniform than the other two tissues. Eighty endophytic fungi selected from 27 genera were evaluated for their inhibition activity against highly virulent Verticillium dahliae isolate Vd080 in vitro. Thirty-nine isolates exhibited fungistasis against the pathogen at varying degrees. Seven species, having high growth inhibition rates (≥75%), exhibited strong antifungal activity against V. dahliae. The antifungal activity of both volatile and nonvolatile metabolites was also investigated. The nonvolatile substances produced by CEF-818 (Penicillium simplicissimum), CEF-325 (Fusarium solani), CEF-714 (Leptosphaeria sp.), and CEF-642 (Talaromyces flavus) completely inhibited V. dahliae growth. These findings deepen our understanding of cotton-endophyte interactions and provide a platform for screening G. hirsutum endophytes with biocontrol potential.

Dynamic behavior analysis of push-the-bit rotary steerable bottom hole assembly

Zhi-chuan Guan,Heng Wang,Yu-cai Shi,Wei-qing Chen,Guo-shan Zhao,Jian-yun Wang,Guang-qiang Cao 대한기계학회 2019 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.33 No.4

Rotary steerable drilling technology is currently an advanced technology in the field of drilling engineering. A good understanding of dynamic behavior of rotary steerable system will benefit the drilling progress. Aimed at push-the-bit type of rotary steerable tool, finite element method was employed to model the rotary steerable bottom hole assembly (RSBHA) and establish the motion equation. Contact interaction between drill string and wellbore wall was analyzed and taken as the constraint condition in solving RSBHA motion equation. Force applied by steering pads was as the external force acting at the pads node. Then RSBHA motion state and bit lateral force were obtained by using Newmark method to solve the motion equation. A case was given and effects of influencing factors were analyzed. The results show that stick-slip phenomenon occurs under 60 r/min rotary speed condition and disappears when rotary speed rises to 90 r/min. Fully developed backward whirl comes into being at the near-bit stabilizer when rotary speed increases to 120 r/min. High weight on bit intensifies bit torsional vibration while larger pads’ steering force decreases the bit torsional vibration. Upper stabilizer plays a part in weakening RSBHA backward whirl under high rotary speed condition but reduces the inclination force. Time average of bit lateral force is mainly determined by pads’ steering force. Based on the results, corresponding suggestions were proposed to make the RSBHA be better used in the field applications.

Knockdown of Ezrin by RNA Interference Reverses Malignant Behavior of Human Pancreatic Cancer Cells in Vitro

Zhong, Zhi-Qiang,Song, Mao-Min,He, Ying,Cheng, Shi,Yuan, Hui-Sheng Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Background: Pancreatic cancer is one of the most aggressive tumors with a dismal prognosis. The membrane cytoskeletal crosslinker Ezrin participates in several functions including cell proliferation, adhesion, motility and survival. There is increasing evidence that Ezrin is overexpressed in vast majority of malignant tumors and regulates tumor progression. However, its roles in pancreatic cancer remain elusive. Methods: Three pairs of specific Ezrin siRNAs were designed and synthetized and screened to determine the most efficient one for construction of a hairpin RNA plasmid targeting Ezrin. After transfection into the Panc-1 pancreatic cancer cell line, real-time quantitative PCR and Western blotting were performed to examine the expression of mRNA and protein. The MTT method was applied to examine the proliferation and the drug sensibility to Gemcitabine. Flow cytometry was used to assess the cycle and apoptosis, while capacity for invasion was determined with transwell chambers. Furthermore, we detected phosphorylated-Erk1/2 protein and phosphorylated-Akt protein by Western blotting. Results: Real-time quantitative PCR and Western blotting revealed that Ezrin expression was notably down-regulated at both mRNA and protein levels by RNA interference (P< 0.01). Proliferation was inhibited and drug resistance to gemcitabine was improved (P< 0.05). Flow cytometry showed that the proportion of cells in the G1/G0 phase increased (P< 0.01), and in G2/M and S phases decreased (P< 0.05), with no apparent differences in apoptosis (P> 0.05). The capacity for invasion was markedly reduced (P< 0.01). In addition, down-regulating Ezrin expression had no effect on phosphorylated-Akt protein (P>0.05), but could decrease the level of phosphorylated-Erk1/2 protein (P< 0.05). Conclusions: RNA interference of Ezrin could inhibit its expression in the pancreatic cancer cells line Panc-1, leading to a potent suppression of malignant behavior in vitro. Assessment of potential as a target for pancreatic cancer treatment is clearly warranted.

Nausea and Vomiting after Transcatheter Arterial Chemoembolization for Hepatocellular Carcinoma: Incidence and Risk Factor Analysis

Wang, Shi-Ying,Zhu, Wen-Hao,Vargulick, Sonya,Lin, Sam Bill,Meng, Zhi-Qiang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.10

Background: Nausea and vomiting after transcatheter arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) are common in clinical practice, but few studies have reported the incidence and risk factors of such events. Objective: The purpose of this study was to analyze the incidence and risk factors of nausea and vomiting after TACE for HCC. Methods: This study was a single-center retrospective analysis of a prospectively maintained database. Between May 2010 and October 2012, 150 patients with HCC were analyzed for incidence and preprocedural risk factors. Results: The incidence of postembolization nausea and vomiting was 38.8% and 20.9%, respectively, in patients with HCC. Patients who developed nausea had lower levels (<100 IU/L) of serum alkaline phosphatase (ALP) compared to those without nausea ($123.04{\pm}69.38$ vs. $167.41{\pm}138.95$, respectively, p=0.044). Female gender correlated to a higher incidence of nausea as well (p=0.024). Patients who developed vomiting, compared to those who did not, also had lower levels (<100 IU/L) of serum ALP ($112.52{\pm}62.63$ vs. $160.10{\pm}127.80$, respectively, p=0.010), and serum alanine transferase (ALT) ($35.61{\pm}22.87$ vs. $4.97{\pm}29.62$, respectively, p=0.045). There were no statistical significances in the incidences of nausea and vomiting between male patients over 50 years old and female patients who have entered menopause (p=0.051 and p=0.409, respectively). Multivariate analysis by logistic regression analysis demonstrated that female gender and ALP>100 IU/L were the most independent predictive factors of postembolization nausea (odds ratio (OR): 3.271, 95% CI: 1.176-9.103, p=0.023 and OR: 0.447, 95% CI: 0.216-0.927, p=0.030, respectively). ALP>100 IU/L was also the most independent predictive risk factor of postembolization vomiting (OR: 0.389, 95% CI: 0.159-0.952, p=0.039). Conclusions: Postembolizaiton nausea and vomiting are common in patients with HCC. Recognition of the risk factors presented above before TACE is important for early detection and proper management of postembolization nausea and vomiting. Nevertheless, future studies are required.

Expression of TS, RRM1, ERCC1, TUBB3 and STMN1 Genes in Tissues of Non-small Cell Lung Cancer and its Significance in Guiding Postoperative Adjuvant Chemotherapy

Zou, Zhi-Qiang,Du, Yi-Ying,Sui, Gang,Xu, Shi-Ning Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.8

Background: To explore the expression of TS, RRM, ERCC1, TUBB3 and STMN1 genes in the tissues of patients with non-small cell lung cancer (NSCLC) and its significance in guiding the postoperative adjuvant chemotherapy. Materials and Methods: Real time polymerase chain reaction (PCR) was applied to detect the expression of TS, RRM, ERCC1, TUBB3 and STMN1 genes in the tissues of NSCLC patients so as to analyze the relationship between the expression of each gene and the clinical characteristics and to guide the postoperative individualized chemotherapy according to the detection results of NSCLC patients. Results: Expression of TS gene was evidently higher in patients with adenocarcinoma than those with non-adenocarcinoma (P=0.013) and so was the expression of ERCC1 (P=0.003). The expression of TUBB3 gene was obviously higher in NSCLC patients in phases I/II and IV than those in phase III ($P_1=0.021$; $P_2=0.004$), and it was also markedly higher in patients without lymph node metastasis than those with (P=0.008). The expression of STMN1 gene was apparently higher in patients in phase I/II than those in phase IV (P=0.002). There was no significant difference between the rest gene expression and the clinical characteristics of NSCLC patients (P>0.05). Additionally, the diseasefree survival (DFS) was significantly longer in patients receiving gene detections than those without (P=0.021). Conclusions: The selection of chemotherapeutic protocols based singly on patients' clinical characteristics has certain blindness. However, the detection of tumor-susceptible genes can guide the postoperative adjuvant chemotherapy and prolong the DFS of NSCLC patients.

A Novel 7-O-Modified Genistein Derivative with Acetylcholinesterase Inhibitory Effect, Estrogenic Activity and Neuroprotective Effect

Da-Hua Shi,Jun-Hua Wu,Zhi-Qiang Yan,Li-Na Zhang,Yu-Rong Wang,Chun-Ping Jiang 대한약학회 2012 Archives of Pharmacal Research Vol.35 No.9

To find the multi-target-directed compounds for the treatment of Alzheimer’s disease (AD), we synthesized 7-(4-(diethylamino)butoxy)-5-hydroxy-3-(4-hydroxyphenyl)-4H-chromen-4-one, a novel 7-O-modified genistein derivative (GS-14), and investigated its acetylcholinesterase (AChE) inhibitory effect, estrogenic activity and neuroprotective effect. GS-14 acted as a selective AChE inhibitor in vitro, with an IC50 value of 0.17 μM and showed no inhibition activity against butyrylcholinesterase (BuChE). The Lineweaver–Burk plot revealed that GS-14 was a non-competitive AChE inhibitor with a Ki value of 0.23 μM and the molecular docking model indicated that GS-14 interacted with the peripheral anionic site (PAS) of AChE. The MCF-7 proliferation assay demonstrated that GS-14 possessed estrogenic activity and GS-14 exhibited a high specificity for estrogen receptor β (ERβ) with a dissociation constant (Ki) of 2.86 nM compared with that of 1.01 μM for estrogen receptor α (ERα) in the molecular docking study. GS-14 also possessed a neuroprotective effect and showed the best protective effect against the β-amyloid protein-induced injury on SH-SY5Y cells at a concentration of 1 nM. Considering its AChEinhibition activity, estrogenic activity and neuroprotective effect, GS-14 may be a potential multi-target agent for the treatment of AD.

Controlled Synthesis of Branched Polystyrene via Free Radical Polymerization of Novel Chain Transfer Monomer

Cui-Ping Li,Jia-Qiang Wang,Jun Lin,Yan Shi,Zhi-Feng Fu 한국고분자학회 2011 Macromolecular Research Vol.19 No.8

The free radical polymerization of styrene was carried out in the presence of a new found chain transfer monomer, p-vinyl benzene sulfonyl chloride (VBSC), which possesses both a chain transfer group and a polymerizable double bond. Branched polystyrene was formed during the polymerization, as indicated by multi-peaks gel permeation chromatography (GPC) curves of the products, the increase in the number-average molecular weight and molecular weight distribution along with monomer conversions. The structure of the obtained polystyrene was analyzed by 1H nuclear magnetic resonance (NMR) spectroscopy. The results showed that with increasing VBSC in the feed, the degree of branching and VBSC unit in the copolymer increased and a shortest polystyrene arm arose from the highest VBSC content in the feed, suggesting that the composition and structure of the branched polystyrenes could be tuned by the amount of VBSC in the feed. By tracing the structure change in the copolymer at various stages of polymerization, the main polymerization process can be regarded as the copolymerization of VBSC with styrene first and then chain transfer to polymeric radical to form branched polystyrene. This strategy is facile and less expensive than the other method.

Synthesis, Crystal Structure and Quantum Chemistry of a Novel Schiff Base N-(2,4-Dinitro-phenyl)-N'-(1-phenyl-ethylidene)-hydrazine

Ji, Ning-Ning,Shi, Zhi-Qiang,Zhao, Ren-Gao,Zheng, Ze-Bao,Li, Zhi-Feng Korean Chemical Society 2010 Bulletin of the Korean Chemical Society Vol.31 No.4

A novel Schiff base N-(2,4-dinitro-phenyl)-N'-(1-phenyl-ethylidene)-hydrazine has been synthesized and structurally characterized by X-ray single crystal diffraction, elemental analysis, IR spectra and UV-vis spectrum. The crystal belongs to monoclinic with space group P21/n. The molecules are connected via intermolecular O-$H{\cdots}O$ hydrogen bonds into 1D infinite chains. The crystal structure is consolidated by the intramolecular N-$H{\cdots}O$ hydrogen bonds. weak intermolecular C-$H{\cdots}O$ hydrogen bonds link the molecules into intriguing 3D framework. Furthermore, Density functional theory (DFT) calculations of the structure, stabilities, orbital energies, composition characteristics of some frontier molecular orbitals and Mulliken charge distributions of the title compound were performed by means of Gaussian 03W package and taking B3LYP/6-31G(d) basis set. The time-dependent DFT calculations have been employed to calculate the electronic spectrum of the title compound, and the UV-vis spectra has been discussed on this basis. The results show that DFT method at B3LYP/6-31G(d) level can well reproduce the structure of the title compound.