Chk2 Regulates Cell Cycle Progression during Mouse Oocyte Maturation and Early Embryo Development

Xiao-Xin Dai,Xing Duan,Honglin Liu,최향순,Nam-Hyung Kim,Shao-Chen Sun 한국분자세포생물학회 2014 Molecules and cells Vol.37 No.2

As a tumor suppressor homologue during mitosis, Chk2 is involved in replication checkpoints, DNA repair, and cell cycle arrest, although its functions during mouse oocyte meiosis and early embryo development remain uncertain. We investigated the functions of Chk2 during mouse oocyte maturation and early embryo development. Chk2 exhibited a dynamic localization pattern; Chk2 expression was restricted to germinal vesicles at the germinal vesicle (GV) stage, was associated with centromeres at pro-meta-phase I (Pro-MI), and localized to spindle poles at meta-phase I (MI). Disrupting Chk2 activity resulted in cell cycle progression defects. First, inhibitor-treated oocytes were arrested at the GV stage and failed to undergo germinal vesicle breakdown (GVBD); this could be rescued after Chk2 inhibition release. Second, Chk2 inhibition after oocyte GVBD caused MI arrest. Third, the first cleavage of early embryo development was disrupted by Chk2 inhibi-tion. Additionally, in inhibitor-treated oocytes, checkpoint protein Bub3 expression was consistently localized at cen-tromeres at the MI stage, which indicated that the spindle assembly checkpoint (SAC) was activated. Moreover, disrupting Chk2 activity in oocytes caused severe chromo-some misalignments and spindle disruption. In inhibitor-treated oocytes, centrosome protein γ-tubulin and Polo-like kinase 1 (Plk1) were dissociated from spindle poles. These results indicated that Chk2 regulated cell cycle progression and spindle assembly during mouse oocyte maturation and early embryo development.

Chk2 Regulates Cell Cycle Progression during Mouse Oocyte Maturation and Early Embryo Development

Dai, Xiao-Xin,Duan, Xing,Liu, Hong-Lin,Cui, Xiang-Shun,Kim, Nam-Hyung,Sun, Shao-Chen Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.2

As a tumor suppressor homologue during mitosis, Chk2 is involved in replication checkpoints, DNA repair, and cell cycle arrest, although its functions during mouse oocyte meiosis and early embryo development remain uncertain. We investigated the functions of Chk2 during mouse oocyte maturation and early embryo development. Chk2 exhibited a dynamic localization pattern; Chk2 expression was restricted to germinal vesicles at the germinal vesicle (GV) stage, was associated with centromeres at pro-metaphase I (Pro-MI), and localized to spindle poles at metaphase I (MI). Disrupting Chk2 activity resulted in cell cycle progression defects. First, inhibitor-treated oocytes were arrested at the GV stage and failed to undergo germinal vesicle breakdown (GVBD); this could be rescued after Chk2 inhibition release. Second, Chk2 inhibition after oocyte GVBD caused MI arrest. Third, the first cleavage of early embryo development was disrupted by Chk2 inhibition. Additionally, in inhibitor-treated oocytes, checkpoint protein Bub3 expression was consistently localized at centromeres at the MI stage, which indicated that the spindle assembly checkpoint (SAC) was activated. Moreover, disrupting Chk2 activity in oocytes caused severe chromosome misalignments and spindle disruption. In inhibitor-treated oocytes, centrosome protein ${\gamma}$-tubulin and Polo-like kinase 1 (Plk1) were dissociated from spindle poles. These results indicated that Chk2 regulated cell cycle progression and spindle assembly during mouse oocyte maturation and early embryo development.

Vector form intrinsic finite-element analysis of static and dynamic behavior of deep-sea flexible pipe

Wu, Han,Zeng, Xiaohui,Xiao, Jianyu,Yu, Yang,Dai, Xin,Yu, Jianxing The Society of Naval Architects of Korea 2020 International Journal of Naval Architecture and Oc Vol.12 No.1

The aim of this study was to develop a new efficient strategy that uses the Vector form Intrinsic Finite-element (VFIFE) method to conduct the static and dynamic analyses of marine pipes. Nonlinear problems, such as large displacement, small strain, and contact and collision, can be analyzed using a unified calculation process in the VFIFE method according to the fundamental theories of point value description, path element, and reverse motion. This method enables analysis without the need to integrate the stiffness matrix of the structure, because only motion equations of particles established according to Newton's second law are required. These characteristics of the VFIFE facilitate the modeling and computation efficiencies in analyzing the nonlinear dynamic problem of flexible pipe with large deflections. In this study, a three-dimensional (3-D) dynamical model based on 3-D beam element was established according to the VFIFE method. The deep-sea flexible pipe was described by a set of spatial mass particles linked by 3-D beam element. The motion and configuration of the pipe are determined by these spatial particles. Based on this model, a simulation procedure to predict the 3-D dynamical behavior of flexible pipe was developed and verified. It was found that the spatial configuration and static internal force of the mining pipe can be obtained by calculating the stationary state of pipe motion. Using this simulation procedure, an analysis was conducted on the static and dynamic behaviors of the flexible mining pipe based on a 1000-m sea trial system. The results of the analysis proved that the VFIFE method can be efficiently applied to the static and dynamic analyses of marine pipes.

Rho-GTPase Effector ROCK Phosphorylates Cofilin in Actin-Meditated Cytokinesis During Mouse Oocyte Meiosis

Duan, Xing,Jun-Liu,Dai, Xiao-Xin,Liu, Hong-Lin,Cui, Xiang-Shun,Kim, Nam-Hyung,Wang, Zhen-Bo,Qiang-Wang,Sun, Shao-Chen Society for the Study of Reproduction [etc.] 2014 BIOLOGY OF REPRODUCTION Vol.90 No.2

During oocyte meiosis, a spindle forms in the central cytoplasm and migrates to the cortex. Subsequently, the oocyte extrudes a small body and forms a highly polarized egg; this process is regulated primarily by actin. ROCK is a Rho-GTPase effector that is involved in various cellular functions, such as stress fiber formation, cell migration, tumor cell invasion, and cell motility. In this study, we investigated possible roles for ROCK in mouse oocyte meiosis. ROCK was localized around spindles after germinal vesicle breakdown and was colocalized with cytoplasmic actin and mitochondria. Disrupting ROCK activity by RNAi or an inhibitor resulted in cell cycle progression and polar body extrusion failure. Time-lapse microscopy showed that this may have been due to spindle migration and cytokinesis defects, as chromosomes segregated but failed to extrude a polar body and then realigned. Actin expression at oocyte membranes and in cytoplasm was significantly decreased after these treatments. Actin caps were also disrupted, which was confirmed by a failure to form cortical granule-free domains. The mitochondrial distribution was also disrupted, which indicated that mitochondria were involved in the ROCK-mediated actin assembly. In addition, the phosphorylation levels of Cofilin, a downstream molecule of ROCK, decreased after disrupting ROCK activity. Thus, our results indicated that a ROCK-Cofilinactin pathway regulated meiotic spindle migration and cytokinesis during mouse oocyte maturation.

Profile of disposition, tissue distribution and excretion of the novel anti-human immunodeficiency virus (HIV) agent W-1 in rats

Ying-Yuan Lu,Xiao-Wei Wang,Xin Wang,Wen-Bing Dai,Qiang Zhang,Pu Li,Ya-Qing Lou,Chuang Lu,Jun-Yi Liu,Guo-Liang Zhang 대한약학회 2016 Archives of Pharmacal Research Vol.39 No.7

The purpose of this study was to characterize the disposition, distribution, excretion and plasma protein binding of 6-benzyl-1-benzyloxymethyl-5-iodouracil (W- 1) in rats. Concentrations of W-1 within biological samples were determined using a validated high performance liquid chromatography method. The plasma protein binding of W-1 was examined by equilibrium dialysis method. After oral administration of W-1 (50, 100 and 200 mg/kg, respectively) in self-microemulsifying drug delivery system formulation, the pharmacokinetic parameters of W-1 were as follows: the peak plasma concentrations (Cmax) were 0.42, 1.50 and 2.55 μg/mL, the area under the curve (AUC0-t) were 0.89, 2.27 and 3.96 lg/h mL and the plasma half-life (t1/2) were 5.15, 3.77 and 3.77 h, respectively. Moreover, the prototype of W-1 was rapidly and extensively distributed into fifteen tissues, especially higher concentrations were detected in intestine, stomach and liver, respectively. The plasma protein binding of W-1 in rat, beagle dog and human were in the range of 97.96–99.13 %. This study suggested that W-1 has an appropriate pharmacokinetics in rats, such as rapid absorption, moderate clearance, and rapid distribution to multiple tissues. Those properties provide important information for further development W-1 as an anti-HIV-1 drug candidate.

IGF-1 from Adipose-Derived Mesenchymal Stem Cells Promotes Radioresistance of Breast Cancer Cells

Yang, Hui-Ying,Qu, Rong-Mei,Lin, Xiao-Shan,Liu, Tong-Xin,Sun, Quan-Quan,Yang, Chun,Li, Xiao-Hong,Lu, Wei,Hu, Xiao-Fang,Dai, Jing-Xing,Yuan, Lin Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.23

Purpose: The aim of this study was to investigate effects of adipose-derived mesenchymal stem cells (AMSCs) on radioresistance of breast cancer cells. Materials and Methods: MTT assays were used to detect any influence of AMSC supernatants on proliferation of breast cancer cells; cell migration assays were used to determine the effect of breast cancer cells on the recruitment of AMSCs; the cell survival fraction post-irradiation was assessed by clonogenic survival assay; ${\gamma}$-H2AX foci number post-irradiation was determined via fluorescence microscopy; and expression of IGF-1R was detected by Western blotting. Results: AMSC supernatants promoted proliferation and radioresistance of breast cancer cells. Breast cancer cells could recruit AMSCs, especially after irradiation. IGF-1 derived from AMSCs might be responsible for the radioresistance of breast cancer cells. Conclusions: Our results suggest that AMSCs in the tumor microenvironment may affect the outcome of radiotherapy for breast cancer in vitro.

Inhibitory effects of piceatannol on human cytomegalovirus (hCMV) in vitro

Wang San-Ying,Zhang Jing,Xu Xiao-Gang,Su Hui-Li,Xing Wen-Min,Zhang Zhong-Shan,Jin Wei-Hua,Dai Ji-Huan,Wang Ya-Zhen,He Xin-Yue,Sun Chuan,Yan Jing,Mao Gen-Xiang 한국미생물학회 2020 The journal of microbiology Vol.58 No.8

Human cytomegalovirus (hCMV) is a ubiquitous herpesvirus, which results in the establishment of a latent infection that persists throughout the life of the host and can be reactivated when the immunity is low. Currently, there is no vaccine for hCMV infection, and the licensed antiviral drugs mainly target the viral enzymes and have obvious adverse reactions. Thus, it is important to search for compounds with antihCMV properties. The present study aimed to investigate the suppressive effects of piceatannol on hCMV Towne strain infection and the putative underlying mechanisms using human diploid fibroblast WI-38 cells. Piceatannol supplementation prevented the lytic changes induced by hCMV infection in WI-38 cells. Furthermore, piceatannol suppressed the expression of hCMV immediate-early (IE) and early (E) proteins as well as the replication of hCMV DNA in a dose-dependent manner. Moreover, hCMV-induced cellular senescence was suppressed by piceatannol, as shown by a decline in the senescence-associated β-galactosidase (SA-β-Gal) activity and decreased production of intracellular reactive oxygen species (ROS). p16INK4a, a major senescence-associated molecule, was dramatically elevated by current hCMV infection that was attenuated by pre-incubation with piceatannol in a dose-dependent manner. These results demonstrated that piceatannol suppressed the hCMV infection via inhibition of the activation of p16INK4a and cellular senescence induced by hCMV. Together, these findings indicate piceatannol as a novel and potent anti-hCMV agent with the potential to be developed as an effective treatment for chronic hCMV infection.

Polymorphisms in Genes of the De Novo Lipogenesis Pathway and Overall Survival of Hepatocellular Carcinoma Patients Undergoing Transarterial Chemoembolization

Wu, You-Sheng,Bao, Deng-Ke,Dai, Jing-Yao,Chen, Cheng,Zhang, Hong-Xin,Yang, YeFa,Xing, Jin-Liang,Huang, Xiao-Jun,Wan, Shao-Gui Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.3

Aberrant expression of genes in de novo lipogenesis (DNL) pathway were associated with various cancers, including hepatocellular carcinoma (HCC). Single nucleotide polymorphisms (SNPs) of DNL genes have been reported to be associated with prognosis of some malignancies. However, the effects of SNPs in DNL genes on overall survival of HCC patients receiving transarterial chemoembolization (TACE) treatment are still unknown. In present study, nine SNPs in three genes (ACLY, ACACA and FASN) in DNL pathway were genotyped using the Sequenom iPLEX genotyping system in a hospital-based cohort with 419 HCC patients treated with TACE, and their associations with HCC overall survival were evaluated by Cox proportional hazard regression analysis under three genetic models (additive, dominant and recessive). Although we did not find any significant results in total analysis (all p>0.05), our stratified data showed that SNP rs9912300 in ACLY gene was significantly associated with overall survival of HCC patients with lower AFP level and SNP rs11871275 in ACACA gene was significantly associated with overall survival of HCC patients with higher AFP level. We further identified the significant interactions between AFP level and SNP rs9912300 or rs11871275 in the joint analysis. Conclusively, our data suggest that genetic variations in genes of DNL pathway may be a potential biomarker for predicting clinical outcome of HCC patients treated with TACE.

Environmental Microbiology and Engineering : Microbial Community Dynamics in Batch High-Solid Anaerobic Digestion of Food Waste Under Mesophilic Conditions

( Jing Yi ),( Bin Dong ),( Yong Gang Xue ),( Ning Li ),( Peng Gao ),( Yu Xin Zhao ),( Ling Ling Dai ),( Xiao Hu Dai ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.2

Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g VSremoval. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

A Three-dimensional Biomechanical Model for Numerical Simulation of Dynamic Pressure Functional Performances of Graduated Compression Stocking (GCS)

Liu, Rong,Kwok, Yi-Lin,Li, Yi,Lao, Terence-T,Zhang, Xin,Dai, Xiao-Qun The Korean Fiber Society 2006 Fibers and polymers Vol.7 No.4

The beneficial effects of graduated compression stockings (GCS) in prophylaxis and treatment of venous disorders of human lower extremity have been recognized. However, their pressure functional performances are variable and unstable in practical applications, and the exact mechanisms of action remain controversial. Direct surface pressure measurements and indirect material properties testing are not enough for fully understanding the interaction between stocking and leg. A three dimensional (3D) biomechanical mathematical model for numerically simulating the interaction between leg and GCS in dynamic wear was developed based on the actual geometry of the female leg obtained from 3D reconstruction of MR images and the real size and mechanical properties of the compression stocking prototype. The biomechanical solid leg model consists of bones and soft tissues, and an orthotropic shell model is built for the stocking hose. The dynamic putting-on process is simulated by defining the contact of finite relative sliding between the two objects. The surface pressure magnitude and distribution along the different height levels of the leg and stress profiles of stockings were simulated. As well, their dynamic alterations with time processing were quantitatively analyzed. Through validation, the simulated results showed a reasonable agreement with the experimental measurements, and the simulated pressure gradient distribution from the ankle to the thigh (100:67:30) accorded with the advised criterion by the European committee for standardization. The developed model can be used to predict and visualize the dynamic pressure and stress performances exerted by compression stocking in wear, and to optimize the material mechanical properties in stocking design, thus, helping us understand mechanisms of compression action and improving medical functions of GCS.