Selection and validation of reference genes for RT-qPCR analysis in Sf9 cell line infected by Heliothis virescens ascovirus 3h (HvAV-3h)

Zi-Shu Chen,Ning-Ning Han,Jian Hong Li,Hu Wan,Guo-Hua Huang 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.2

The real-time quantitative reverse transcription PCR (qRT-PCR) is an efficient and pervasive technique for analysis gene expression. Despite the fact that this technique has been extensively used to explore the gene functions, the stability of the reference genes still requires being validated at different experimental conditions. This research aims to confirm that the expression stability of seven candidates reference genes under different concentrations sterile hemolymph containing virus (2, 4, 6, 108-fold) infects the Spodoptera frugiperda cells line (Sf9). The appropriate reference genes as endogenous controls were determined through four computational algorithms (ΔCt, NormFinder, BestKeeper, and geNorm) and an integrated online software RefFinder. According to results of this study, TUB and ACT1 were proved to be the most stable reference genes in E2 fold and E4 fold respectively. PPI was the best reference gene in E6 and E8 fold, 28S and TUB were the most stable pair reference genes through all samples. These results can facilitate basic researches of the molecular mechanism of HvAV- 3h and also prove to be helpful for the standard RT-PCR method.

Isolation of NH4+-Tolerant Mutants of Actinobacillus succinogenes for Succinic Acid Production by Continuous Selection

( Gui Zi Ye ),( Min Jiang ),( Jian Li ),( Ke Quan Chen ),( Yong Lan Xi ),( Shu Wen Liu ),( Ping Wei ),( Ping Kai Ouyang ) 한국미생물 · 생명공학회 2010 Journal of microbiology and biotechnology Vol.20 No.8

Actinobacillus succinogenes, a representative succinic-acid-producing microorganism, is seriously inhibited by ammonium ions, thereby hampering the industrial use of A. succinogenes with ammonium-ion-based materials as the pH controller. Therefore, this study isolated an ammonium-ion-tolerant mutant of A. succinogenes using a continuous-culture technique in which all the environmental factors, besides the stress (ammonium ions), were kept constant. Instead of operating the mutant-generating system as a nutrient-limited chemostat, it was used as a nutrientunlimited system, allowing the cells to be continuously cultured at the maximum specific growth rate. The mutants were isolated on agar plates containing the acid-base indicator bromothymol blue and a high level of ammonium ions that would normally kill the parent strain by 100%. When cultured in anaerobic bottles with an ammonium ion concentration of 354 mmol/l, the mutant YZ0819 produced 40.21 g/l of succinic acid with a yield of 80.4%, whereas the parent strain NJ113 was unable to grow. When using NH4OH to buffer the culture pH in a 3.0 l stirred bioreactor, YZ0819 produced 35.15 g/l of succinic acid with a yield of 70.3%, which was 155% higher than that produced by NJ113. In addition, the morphology of YZ0819 changed in the fermentation broth, as the cells were aggregated from the beginning to the end of the fermentation. Therefore, these results indicate that YZ0819 can efficiently produce succinic acid when using NH4OH as the pH controller, and the formation of aggregates can be useful for transferring the cells from a cultivation medium for various industrial applications.

A Novel Role of Classical Swine Fever Virus E^rns Glycoprotein in Counteracting the Newcastle Disease Virus (NDV)-mediated IFN-β Induction

Xia, Yan-Hua,Chen, Liu,Pan, Zi-Shu,Zhang, Chu-Yu Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.5

$E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

A Novel Role of Classical Swine Fever Virus E(rns) Glycoprotein in Counteracting the Newcastle Disease Virus (NDV)-mediated IFN-β Induction

( Yan Hua Xia ),( Liu Chen ),( Zi Shu Pan ),( Chu Yu Zhang ) 생화학분자생물학회 2007 BMB Reports Vol.40 No.5

Perforation optimization of hydraulic fracturing of oil and gas well

Zhu, Hai Yan,Deng, Jin Gen,Chen, Zi Jian,An, Feng Chen,Liu, Shu Jie,Peng, Cheng Yong,Wen, Min,Dong, Guang Techno-Press 2013 Geomechanics & engineering Vol.5 No.5

Considering the influences of fluid penetration, casing, excavation processes of wellbore and perforation tunnels, the seepage-deformation finite element model of oil and gas well coupled with perforating technique is established using the tensile strength failure criterion, in which the user-defined subroutine is developed to investigate the dynamic evolvement of the reservoir porosity and permeability. The results show that the increases of perforation angle and decreases of perforation density lead to a higher fracture initiation pressure, while the changes of the perforation diameter and length have no evident influences on the fracture initiation pressure. As for initiation location for the fracture in wellbore, it is on the wellbore face while considering the presence of the casing. By contrast, the fractures firstly initiate on the root of the tunnels without considering casing. Besides, the initial fracture position is also related with the perforation angle. The fracture initiation position is located in the point far away from the wellbore face, when the perforation angle is around $30^{\circ}$; however, when the perforation angle is increased to $45^{\circ}$, a plane fracture is initiated from the wellbore face in the maximum horizontal stress direction; no fractures was found around perforation tunnels, when the angel is close to $90^{\circ}$. The results have been successfully applied in an oilfield, with the error of only 1.1% comparing the fracture initiation pressure simulated with the one from on-site experiment.

Hydraulic fracturing experiments of highly deviated well with oriented perforation technique

Zhu, Hai Y.,Deng, Jin G.,Liu, Shu J.,Wen, Min,Peng, Cheng Y.,Li, Ji R.,Chen, Zi J.,Hu, Lian B.,Lin, Hai,Guang, Dong Techno-Press 2014 Geomechanics & engineering Vol.6 No.2

In order to investigate the effect of different perforation angles (the angle between the perforation direction and the maximum horizontal principal stress) on the fracture initiation and propagation during hydraulic fracturing of highly deviated well in oil & gas saturated formation, laboratory experiments of the hydraulic fracturing had been carried out on the basis of non-dimensional similar criteria by using 400^3 $mm^3$ cement cubes. A plane fracture can be produced when the perforations are placed in the direction of the maximum horizontal principal stress. When the perforation angle is $45^{\circ}$, the fractures firstly initiate from the perforations at the upper side of the wellbore, and then turn to the maximum horizontal principal stress direction. When the well deviation angle and perforation angle are both between $45^{\circ}$ and $90^{\circ}$, the fractures hardly initiate from the perforations at the lower side of the wellbore. Well azimuth (the angle between the wellbore axis and the maximum horizontal principal stress) has a little influence on the fracture geometries; however it mainly increases the fracture roughness, fracture continuity and the number of secondary fractures, and also increases the fracture initiation and propagation pressure. Oriented perforating technology should be applied in highly deviated well to obtain a single plane fracture. If the well deviation angle is smaller, the fractures may link up.

Mechanisms of Electroacupuncture Pretreatment in Alleviating Myocardial Ischemia Reperfusion Injury: Interactions between the Cerebellar Fastigial Nucleus and Lateral Hypothalamic Area

Yu Qing,Wu Li-bin,Zhang Fan,Wei Xiao-tong,Chen Pian-pian,Wang Shuai-ya,Cai Mei-yi,Shu Qi,Li Liao-yuan,Wu Zi-jian,Cai Rong-lin,Hu Ling 사단법인약침학회 2021 Journal of Acupuncture & Meridian Studies Vol.14 No.6

Background: Myocardial ischemia reperfusion injury (MIRI) is an important mechanism of post-myocardial infarction injury and a main cause of death in patients with ischemic heart disease. Electroacupuncture (EA) pretreatment is effective for the prevention and treatment of MIRI, but mechanisms mediating the effects of cardiovascular disease EA treatments remain unclear. Objectives: To determine whether the lateral hypothalamus (LHA) and the cerebellar fastigial nucleus (FN) are involved in the protective effects of EA stimulation on MIRI. Methods: EA pretreatment was performed for 7 days before the establishment of the MIRI model. ST-segment changes on electrocardiograms were recorded and the Curtis–Walker arrhythmia score was used to evaluate changes in reperfusion injury. Hematoxylin–eosin staining was applied to evaluate the pathological and morphological changes in myocardial tissue. c-fos expression in the LHA and FN was determined by immunofluorescence staining. Glutamic (Glu) and γ-Aminobutyric acid (GABA) levels were measured using a high-performance liquid chromatography-electrochemical method. Results: EA pretreatment reduced ST-segment elevation, arrhythmia scores, and morphological changes in MIRI myocardial cells in rats, and decreased the c-fos protein expression in LHA/FN nuclei. MIRI was associated with an imbalance between GABA and Glu levels, whereas EA pretreatment increased GABA levels and decreased Glu levels in the LHA/FN. Conclusion: FN and LHA are involved in the EA-mediated attenuation of MIRI. Pretreatment with EA plays a protective role in the myocardium by regulating Glu and GABA release in the LHA and FN.

Association between Pax8-PPARγ1 Rearrangement and Follicular Thyroid Cancer: a Meta-Analysis

Li, Hang-Yu,Xie, Zhi-Hao,Xu, Cong-Hui,Pu, Mei-Ling,Chen, Zi-Yan,Yu, Miao,Wang, Heng-Shu,Zhou, Chen-Ming,Pu, Chao-Yu,Liu, Wei Asian Pacific Journal of Cancer Prevention 2016 Asian Pacific journal of cancer prevention Vol.17 No.9

Background: Pax8 and peroxisome proliferator-activated receptor gamma 1 gene (Pax8-$PPAR{\gamma}1$) are important factors in tumors. Several studies have suggested that follicular thyroid cancer may arise from Pax8- $PPAR{\gamma}1$ rearrangement. In order to have a better understanding of the association between Pax8-$PPAR{\gamma}1$ rearrangement and follicular thyroid cancer, we conducted the presenmt meta-analysis. Materials and Methods: The information was extracted from PubMed, EMBASE and Web of Science. Statistic analysis was performed with Stata12.0 software. Odds ratios (ORs) were calculated using a fixed-effects model. We also performed heterogeneity and publication bias analyses. Results: Nine studies including 198 follicular thyroid cancer patients and 268 controls were considered eligible. The frequency of Pax8-$PPAR{\gamma}1$ rearrangement was significantly higher in the follicular thyroid cancer group than in the control group, with a pooled OR of 6.63 (95%CI=3.50-12.7). In addition, through subgroup analysis, the OR between Pax8-$PPAR{\gamma}1$ rearrangement and follicular thyroid cancer was 6.04 (95%CI = 3.18-11.5) when using benign tumor tissues as controls. The OR for the method subgroup was 9.99 (95% CI =4.86-20.5) in the RT-PCR. Conclusions: The final results demonstrated that Pax8-$PPAR{\gamma}1$ rearrangement has significant association with follicular thyroid cancer.