FKBP3 aggravates the malignant phenotype of diffuse large B-cell lymphoma by PARK7-mediated activation of Wnt/ß-catenin signalling.

Diffuse large B-cell lymphoma (DLBCL) is difficult to treat due to the high recurrence rate and therapy intolerance, so finding potential therapeutic targets for DLBCL is critical. FK506-binding protein 3 (FKBP3) contributes to the progression of various cancers and is highly expressed in DLBCL, but the role of FKBP3 in DLBCL and its mechanism are not clear. Our study demonstrated that FKBP3 aggravated the proliferation and stemness of DLBCL cells, and tumour growth in a xenograft mouse model. The interaction between FKBP3 and parkinsonism associated deglycase (PARK7) in DB cells was found using co-immunoprecipitation assay. Knockdown of FKBP3 enhanced the degradation of PARK7 through increasing its ubiquitination modification. Forkhead Box O3 (FOXO3) belongs to the forkhead family of transcription factors and inhibits DLBCL, but the underlying mechanism has not been reported. We found that FOXO3 bound the promoter of FKBP3 and then suppressed its transcription, eventually weakening DLBCL. Mechanically, FKBP3 activated Wnt/ß-catenin signalling pathway mediated by PARK7. Together, FKBP3 increased PARK7 and then facilitated the malignant phenotype of DLBCL through activating Wnt/ß-catenin pathway. These results indicated that FKBP3 might be a potential therapeutic target for the treatment of DLBCL.

Effect of seat back angle on preferred seat pan inclination for the development of highly automated vehicles.

Recent studies on occupants' safety in reclined positions suggest that a more inclined seat pan could be needed to reduce the occurrence of submarining. This study aimed to investigate whether a more inclined seat pan would also be comfortable for occupants. Eighteen volunteers participated in the experiment. They were asked to self-select seat pan inclination for seat back angles from 20 to 60 degrees using a reconfigurable experimental seat from two initial seat pan angles (10 and 40 degrees from the horizontal). On average, preferred seat pan angle varied from 11.3(±2.1, standard deviation) to 29.9(±6.8), 12.5(±3.8) to 37.4(±3.7), and 12.8(±4.8) to 38.6(±2.7) degrees for seat pan angles of 20, 40, and 60 degrees respectively. The shear force analysis suggests that the seat pan inclination might be self-selected to reduce the forward shear, while a high inclination angle with a noticeable backward shear was also preferred.Practitioner summary: Preferred range of seat pan inclination for different seat back angles studied for the development of highly automated vehicles. The present work provides quantitative guidelines for specifying comfortable seating in a reclined position.

Production of all-male non-transgenic zebrafish by conditional primordial germ cell ablation.

Many fish species exhibit remarkable sexual dimorphism, with males possessing numerous advantageous traits for commercial production by aquaculture such as faster growth rate, more efficient food energy utilization for muscle development, and better breeding performance. Several studies have shown that a decrease in the number of primordial germ cells (PGCs) during early development leads predominantly to male progeny. In this study, we developed a method to obtain all-male zebrafish (Danio rerio) by targeted PGC ablation using the nitroreductase/metronidazole (NTR/Mtz) system. Embryos generated by female heterozygous Tg(nanos3:nfsB-mCherry-nanos3 3'UTR) and male wild-types (WTs) were treated with vehicle or Mtz. Compared to vehicle-treated controls, 5.0 and 10.0 mM Mtz treatment for 24 h significantly reduced the number of PGCs and yielded an exclusively male phenotype in adulthood. The gonads of offspring treated with 5.0 mM Mtz exhibited relatively normal morphology and histological characteristics. Furthermore, these males were able to chase females, spawn, and produce viable offspring, while about 20.0% of males treated with 10.0 mM Mtz were unable to produce viable offspring. The 5.0 mM Mtz treatment protocol may thus be suitable for large-scale production of fertile male offspring. Moreover, about half of these males were WT as evidenced by the absence of nfsB gene expression. It may thus be possible to breed an all-male WT fish population by Mtz-mediated PGC ablation.

Comparative clinical study of short-term outcomes between table fenestrated and chimney endovascular aneurysm repair for hostile neck aneurysms.

OBJECTIVES: Hostile neck abdominal aortic aneurysm (AAA) is challenging for standard endovascular aneurysm repair (EVAR). We sought to compare fenestrated endovascular aneurysm repair (fEVAR) and chimney endovascular aneurysm repair (chEVAR) for hostile neck AAA. METHODS: Patients were identified retrospectively. Hostile neck anatomy was defined as a proximal neck length of <15 mm or angulation >60°. The choice of fEVAR or chEVAR was based on neck anatomy and physician preference. Type I endoleak (T1EL) was the primary outcome. Other outcomes included type III endoleak (T3EL), visceral stent occlusion, renal insufficiency, reintervention, and mortality. RESULTS: A total of 84 patients were included from April 2012 to December 2021. fEVAR and chEVAR patients were 48 and 36 cases, respectively. Both groups showed similar rate of T1EL, T3EL, visceral stent occlusion, renal insufficiency, reintervention, and mortality. However, chEVAR patients had a more tortuous neck (61.1% vs. 16.7%, p < 0.001), while fEVAR patients had a greater neck size (29.5 ± 6.3 mm vs. 24.5 ± 4.8 mm, p < 0.001) and more reconstructing target arteries (2.2 ± 1.1 vs 1.3 ± 0.6, p < 0.001). CONCLUSIONS: fEVAR and chEVAR show similar safe and effective outcomes in well-selected hostile neck. fEVAR might be able to reconstruct multiple visceral arteries, and chEVAR seems justified in patients with poor anatomical suitability for fEVAR.

Division behaviours and their effects on pre-implantation development of pig embryos.

The quality of pre-implantation embryos could affect developmental efficiency after embryo transfer. However, the assessment of pre-implantation embryos was unsatisfactory, especially in pig embryos to date. Therefore, this study was designed to investigate available and applicable parameters that indicate developmental potential and quality of porcine pre-implantation embryos produced by handmade cloning (HMC), and parthenogenetic activation without zona pellucida (PAZF) and with zona pellucida (PAZI). Firstly, a common division behaviour was detected, that is the formation of uneven division with two unequal size blastomeres (UD 2-cell), especially in HMC embryos; then, the proportion of UD 2-cell was found to be significantly higher than that of even division with equal size blastomeres (ED 2-cell) (72.56 ± 4.56 vs. 24.57 ± 1.92). The formation of UD 2-cell might be due to the spindle migration along the long axis in 1-cell stage, and the cleavage furrow was not formed in the centre of cytoplasm. In the two sister blastomeres of UD 2-cell, uneven distribution of organelles (mitochondria and lipid droplet) was observed with lower proportion in the smaller one (p < .05). Although no difference in blastocyst rate was observed between UD and ED 2-cell embryos, the cell number per blastocyst from UD 2-cell embryos was lower than that from ED 2-cell embryos (44.15 ± 2.05 vs. 51.55 ± 1.83). Besides, because of non-synchronized division of each blastomere, the following three cleavage routes were observed in all HMC/PAZF/PAZI embryos: T1 (2-cell → 3-cell → 4-cell → ≥5-cell → morula → blastocyst), T2 (2-cell → 3-cell → 4-cell → morula → blastocyst) and T3 (2-cell → 3-cell/4-cell → morula → blastocyst). Therefore, in pig in vitro-produced embryos, division behaviours of uneven volume of cytoplasm and non-synchronized cell cycles were observed at the early embryonic developmental stage, which might be another potential factor to evaluate embryonic development.

Determinants of ecological and carbon footprints to assess the framework of environmental sustainability in BRICS countries: A panel ARDL and causality estimation model.

The purpose of the present study is to evaluate the effects of financial development, human capital, globalization, and renewable energy consumption on ecological and carbon footprint. For this purpose, the data was collected from four countries i.e. Brazil, Russia, India, and China for 20 years (1997-2016). The 20 years panel data collected for these countries was analyzed through "auto-regressive distributed lag" (ARDL). Findings of this study reveal that human capital is a significant positive determinant of environmental degradation in short run and long run while renewable energy consumption is a significant negative determinant of environmental degradation in short run and long run. Findings further reveal that globalization significantly reduces environmental degradation in short run and long run. Furthermore, findings show that the financial development significantly positively affects the environmental degradation in short run and long run. Therefore, renewable energy consumption and globalization are found to be negative predictors of ecological footprint and carbon footprint while human capital and financial development are found to be positive predictors of ecological footprint and carbon footprint. These findings are expected to be great contribution to the literature and practice, as they will help researchers and policymakers to realize the relative importance of energy, educational, economic, and trade policies in determining the environmental degradation.

Compound essential oils relieve oxidative stress caused by PM2 .5 exposure by inhibiting autophagy through the AMPK/mTOR pathway.

Fine particulate matter (PM2.5 ) potentially damages the respiratory system and causes respiratory diseases. Compound essential oils (CEOs) have been shown to alleviate the damage to the lung and macrophages caused by PM2.5 . However, the effect of PM2.5 exposure on the brain has rarely been investigated. When oxidative stress occurs in the brain, it readily causes neurological diseases. Autophagy is intimately involved in many physiological processes, especially processes important for the brain. Blocked or excessive autophagy causes a series of brain diseases, such as cerebral ischemia and stroke. This study investigated whether CEOs regulate excessive autophagy and reduce the oxidative stress caused by PM2.5 in the brain and BV2 microglial cells. PM2.5 increased the levels of ROS, Nox2, NF-κB and MDA while decreasing superoxide dismutase and HO-1 levels, which led to oxidative stress in the brain. The increased LC3 level and decreased P62 level suggested that PM2.5 exposure increased the level of autophagy. After exposure to PM2.5 , the levels of 5'-adenosine monophosphate-activated protein kinase (AMPK) increased, while the levels of mammalian target of rapamycin (mTOR) decreased, suggesting that PM2.5 might induce autophagy by activating the AMPK/mTOR pathway. In addition, CEOs alleviated oxidative stress and autophagy induced by PM2.5 . Therefore, we concluded that CEOs reduce oxidative stress induced by PM2.5 exposure by inhibiting autophagy via the AMPK/mTOR signaling pathway, and these findings provide new opportunities for the prevention of PM2.5 -induced brain diseases.

Exploration of Driver Posture Monitoring Using Pressure Sensors with Lower Resolution.

Pressure sensors are good candidates for measuring driver postural information, which is indicative for identifying driver's intention and seating posture. However, monitoring systems based on pressure sensors must overcome the price barriers in order to be practically feasible. This study, therefore, was dedicated to explore the possibility of using pressure sensors with lower resolution for driver posture monitoring. We proposed pressure features including center of pressure, contact area proportion, and pressure ratios to recognize five typical trunk postures, two typical left foot postures, and three typical right foot postures. The features from lower-resolution mapping were compared with those from high-resolution Xsensor pressure mats on the backrest and seat pan. We applied five different supervised machine-learning techniques to recognize the postures of each body part and used leave-one-out cross-validation to evaluate their performance. A uniform sampling method was used to reduce number of pressure sensors, and five new layouts were tested by using the best classifier. Results showed that the random forest classifier outperformed the other classifiers with an average classification accuracy of 86% using the original pressure mats and 85% when only 8% of the pressure sensors were available. This study demonstrates the feasibility of using fewer pressure sensors for driver posture monitoring and suggests research directions for better sensor designs.

Influences of Extraction Methods on Physicochemical and Functional Characteristics of Three New Bulbil Starches from Dioscorea opposita Thunb. cv. Tiegun.

Starches from the bulbils of Dioscoreae opposita Thunb. cv. Tiegun were isolated by aqueous steeping (SBS), enzyme extraction (EBS), and alkaline extraction (ABS) methods, respectively. The physicochemical, mineral composition, thermal and morphological characteristics of these starches were investigated. The starch granules were oval, spherical and kidney-shaped and its crystal type is a mixture of A-type and B-type patterns. The starches having larger average granule size showed more amylose and phosphorus contents than those with smaller average granule size. Differential scanning calorimetry (DSC) showed that the SBS had an endothermic transition ranging from 65.8 °C to 76.3 °C with an enthalpy of 2.0 J/g. The endothermic transitions of ABS and EBS showed the regions of 67.9 °C to 73.0 °C, and 66.8 °C to 82.0 °C, respectively. The gelationization enthalpies of ABS and EBS were 13.8 and 11.5 J/g, respectively. Additionally, ABS presented greater clarity in comparison with EBS and SBS. Pasting properties indicated that ABS had the highest peak viscosity, breakdown, but SBS had the lowest trough, final viscosity, setback, and pasting temperature. Generally, ABS and EBS could be used as food thickener or frozen food additives. SBS and EBS were potential technological alternatives in quality preservation of frozen starch-based products and other industrial applications.

Maximal isometric force exertion predicted by the force feasible set formalism: application to handbraking.

The aim of this study was to test the capacity of the force feasible set formalism to predict maximal force exertion during isometric handbraking. Maximal force exertion and upper-limb posture were measured with a force sensor embedded in a handbrake and an optoelectronic system, respectively. Eleven subjects participated in the experiment which consisted of exerting the maximal force in isometric conditions considering five hand brake positions relative to the seat H-point. Then, maximal force was predicted by the force feasible set obtained from an upper-limb musculoskeletal model. The root-mean-square (RMS) of the angle between measured and predicted forces was 8.4° while the RMS error (RMSE) for amplitude prediction was 95.4 N. However, predicted, and measured force amplitudes were highly correlated (r = 0.88, p < 0.05, slope = 0.97, intercept = 73.3N) attesting the capacity of the model to predict force exertion according to the subject's posture. The implications in the framework of ergonomics are then discussed. Practitioner summary: Maximal force exertion is of paramount importance in digital human modelling. We used the force feasible set formalism to predict maximal force exertion during handbraking from posture and anthropometric data. The predicted and measured force orientation showed a RMS of 8.4° while amplitude presented a RMSE of 95.4 N with a strong correlation (r = 0.88, p < 0.05, slope 0.97, intercept 77.3 N).

A parametric investigation on seat/occupant contact forces and their relationship with initially perceived discomfort using a configurable seat.

The present work investigates the contact forces between sitters and seat as well as their correlations with perceived discomfort. Twelve different economy class aeroplane seat configurations were simulated using a multi-adjustable experimental seat by varying seat pan and backrest angles, as well as seat pan compressed surface. Eighteen males and 18 females, selected by their body mass index and stature, tested these configurations for two sitting postures. Perceived discomfort was significantly affected by seat parameters and posture and correlated both with normal force distribution on the seat-pan surface and with normal forces at the lumbar and head supports. Lower discomfort ratings were obtained for more evenly distributed normal forces on the seat pan. Shear force at the seat pan surface was at its lowest when sitters were allowed to self-select their seat-pan angle, supporting that a shear force should be reduced but not zeroed to improve seating comfort. Practitioner Summary: The effects of seat-pan and backrest angle, anthropometric dimensions and sitting posture on contact forces and perceived discomfort were investigated using a multi-adjustable experimental seat. In addition to preferred seat profile parameters, the present work provides quantitative guidelines on contact force requirement for improving seating comfort.

Binary Two-Dimensional Honeycomb Lattice with Strong Spin-Orbit Coupling and Electron-Hole Asymmetry.

Two-dimensional (2D) materials consisting of heavy atoms with particular arrangements may host exotic quantum properties. Here, we report a unique 2D semiconducting binary compound, a Sn_{2}Bi atomic layer on Si(111), in which hexagons are formed by bonding Bi with a triangular network of Sn. Because of the unique honeycomb configuration, the heavy elements, and the energy-dependent hybridization between Sn and Bi, 2D Sn_{2}Bi not only shows strong spin-orbit coupling effects but also exhibits high electron-hole asymmetry: Nearly free hole bands and dispersionless flat electron bands coexist in the same system. By tuning the Fermi level, it is possible to preserve both nearly free and strongly localized charge carriers in the same 2D material, which provides an ideal platform for the studies of strongly correlated phenomena and possible applications in nanodevices.

Effects of Anthropometric Variables and Seat Height on Automobile Drivers' Preferred Posture With the Presence of the Clutch.

Objective The effects of seat height and anthropometric dimensions on drivers' preferred postures were investigated using a multiadjustable vehicle mock-up with a large number of adjustments and extended ranges. Background Many studies have been conducted on preferred driving posture under different test conditions showing mixed and even contradictory findings. No studies thus far have considered the clutch and compared Chinese and European drivers. Method Four seat height conditions were tested: free and three imposed heights (250, 300, and 350 mm). Sixty-one subjects (40 French-born and 21 Chinese-born) participated in the experiment, covering a large range of stature and sitting height-to-stature ratio. The RAMSIS kinematic model was used to reconstruct postures, and main intersegmental angles were extracted for characterizing posture. Results Under the free seat height condition, no significant differences in preferred intersegmental angles were observed between different participant groups. Seat height mainly affected trunk-thigh angle, whereas it had almost no effect on trunk orientation and other intersegmental angles. Chinese participants sat more forward in the seat, leading to a more opened trunk-thigh angle and a more reclined trunk. Conclusions Results suggest that intersegmental angles of preferred posture are not dependent on anthropometric dimensions, although shorter drivers prefer a slightly less reclined trunk. Self-selected driving posture results from a compromise between maintaining the intersegmental angles in one's preferred range and a preferred trunk orientation in space. Applications The findings contribute to a better understanding of preferred driving postures and would be helpful for improving vehicle interior design.

Beneficial Effects of Melatonin on the In Vitro Maturation of Sheep Oocytes and Its Relation to Melatonin Receptors.

(1) Background: The binding sites of melatonin, as a multifunctional molecule, have been identified in human, porcine, and bovine samples. However, the binding sites and mechanisms of melatonin have not been reported in sheep; (2) Methods: Cumulus-oocyte complexes (COCs) were cultured in TCM-199 supplemented with melatonin at concentrations of 0, 10-3, 10-5, 10-7, 10-9, and 10-11 M. Melatonin receptors (MT1 and MT2) were evaluated via immunofluorescence and Western blot. The effects of melatonin on cumulus cell expansion, nuclear maturation, embryo development, and related gene (GDF9, DNMT1, PTX3, HAS2, and EGFR) expression were investigated. The level of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were evaluated in oocytes and cumulus, respectively; (3) Results: Both MT1 and MT2 were expressed in oocytes, cumulus cells, and granulosa cells. Melatonin with a concentration of 10-7 M significantly enhanced the rates of nuclear maturation, cumulus cells expansion, cleavage, and blastocyst. Melatonin enhanced the expression of BMP15 in oocytes and of PTX3, HAS2, and EGFR in cumulus cells. Melatonin decreased the cAMP level of oocytes but enhanced the cGMP level in oocytes and cumulus cells; (4) Conclusion: The higher presence of MT1 in GV cumulus cells and the beneficial effects of melatonin indicated that its roles in regulating sheep oocyte maturation may be mediated mainly by the MT1 receptor.

Involvement of G9A-like protein (GLP) in the development of mouse preimplantation embryos in vitro.

G9A-like protein (GLP) plays an important role in mouse early embryonic development. Glp-deficient embryos exhibit severe growth retardation and defects that lead to lethality at approximately Embryonic Day 9.5. In the present study we investigated the effect of microinjection of Glp-specific short interference (si) RNA into mouse zygotes on in vitro embryonic development. Knockdown of Glp induced abnormal embryonic development and reduced blastocyst formation. Expression of the pluripotency markers octamer-binding transcription factor 4 (Oct4), SRY (sex determining region Y)-box 2 (Sox2) and Nanog was also significantly decreased in Glp-deficient embryos. The apoptotic index and expression of two pro-apoptotic genes, namely Caspase 3 and Caspase 9, were increased in Glp-deficient embryos. Moreover, methylation levels of dimethylated H3K9 (H3K9me2) were decreased in Glp-knockdown embryos. In conclusion, the results of the present study suggest that Glp deficiency suppresses H3K9me2 modification and hinders mouse embryo development in vitro.

Qualitative and Quantitative Analysis of Lignan Constituents in Caulis Trachelospermi by HPLC-QTOF-MS and HPLC-UV.

A high-performance liquid chromatography coupled with quadrupole tandem time-of-flight mass (HPLC-QTOF-MS) and ultraviolet spectrometry (HPLC-UV) was established for simultaneous qualitative and quantitative analysis of the major chemical constituents in Caulis Trachelospermi, respectively. The analysis was performed on an Agilent Zorbax Eclipse Plus C18 column (4.6 mm×150 mm, 5 µm) using a binary gradient system of water and methanol, with ultraviolet absorption at 230 nm. Based on high-resolution ESI-MS/MS fragmentation behaviors of the reference standards, the characteristic cleavage patterns of lignano-9, 9'-lactones and lignano-8'-hydroxy-9, 9'-lactones were obtained. The results demonstrated that the characteristic fragmentation patterns are valuable for identifying and differentiating lignano-9,9'-lactones and lignano-8'-hydroxy-9,9'-lactones. As such, a total of 25 compounds in Caulis Trachelospermi were unambiguously or tentatively identified via comparisons with reference standards or literature. In addition, 14 dibenzylbutyrolatone lignans were simultaneously quantified in Caulis Trachelospermi by HPLC-UV method. The method is suitable for the qualitative and quantitative analyses of dibenzylbutyrolatone lignans in Caulis Trachelospermi.

An assessment of the realism of digital human manikins used for simulation in ergonomics.

In this study, the accuracy of the joint centres of the manikins generated by RAMSIS and Human Builder (HB), two digital human modelling (DHM) systems widely used in industry for virtual ergonomics simulation, was investigated. Eighteen variously sized females and males were generated from external anthropometric dimensions and six joint centres (knee, hip and four spine joints) were compared with their anatomic locations obtained from the three-dimensional reconstructed bones from a low-dose X-ray system. Both RAMSIS and HB could correctly reproduce external anthropometric dimensions, while the estimation of internal joint centres location presented an average error of 27.6 mm for HB and 38.3 mm for RAMSIS. Differences between both manikins showed that a more realistic kinematic linkage led to better accuracy in joint location. This study opens the way to further research on the relationship between the external body geometry and internal skeleton in order to improve the realism of the internal skeleton of DHMs, especially for a biomechanical analysis requiring information of joint load and muscle force estimation. PRACTITIONER SUMMARY: This study assessed two digital human modelling (DHM) systems widely used in industry for virtual ergonomics. Results support the need of a more realistic human modelling, especially for a biomechanical analysis and a standardisation of DHMs.

Three-Dimensional Rotations of the Scapula During Arm Abduction: Evaluation of the Acromion Marker Cluster Method in Comparison With a Model-Based Approach Using Biplanar Radiograph Images.

Noninvasive methods enabling measurement of shoulder bone positions are paramount in clinical and ergonomics applications. In this study, the acromion marker cluster (AMC) method is assessed in comparison with a model-based approach allowing scapula tracking from low-dose biplanar radiograph images. Six healthy male subjects participated in this study. Data acquisition was performed for 6 arm abduction positions (0°, 45°, 90°, 120°, 150°, 180°). Scapula rotations were calculated using the coordinate systems and angle sequence was defined by the ISB. The comparison analysis was based on root mean square error (RMSE) calculation and nonparametric statistical tests. RMSE remained under 8° for 0° to 90° arm abduction and under 13.5° for 0° to 180° abduction; no significant differences were found between the 2 methods. Compared with previous works, an improved accuracy of the AMC approach at high arm abduction positions was obtained. This could be explained by the different sources of data used as the "gold standard."

[Fragmentation pathways of five furocoumarine using line ion trap with orbitrapmass spectrometry].

OBJECTIVE: The multiple levels fragmentations of five furocoumarine (psoralen, xanthotoxin, bergapten, oxypeucedanin, and byakangelicol) in Angelica dahurica have been demonstrated using LTQ-Orbitrap mass spectrometry with high resolution and high mass accuracy to discover the possible,fragmentation regularity. METHOD: Duringcollsion-induced dissociation (CID), the MS(n) data of the five compoundswhich were gained in the positive ion mode at 35ev collision energy by direct injection syrings method were analyzed using Xcalibar 2.0 Software to infer the formula of these fragmentations. RESULT: The results indicated that the five compounds have similar fragmentation process with CO meutral lost at C5,C8-subsituents and furan ring, meanwhile the meutralloss of CO2 occurred easily at lactone group. CONCLUSION: This method is helpful in identifying the structures of other furocoumarinein Angelica dahuricaand their metabolites in vivo.

EZH2 is essential for development of mouse preimplantation embryos.

Enhancer of zeste homologue 2 (Ezh2) is essential for the development of the early mouse preimplantation embryo. Loss of Ezh2 results in embryonic lethality in mice. Ezh2-deficient embryos display impaired outgrowth potential, defective establishment of Ezh2-null embryonic stem (ES) cells and adherence and differentiation of the trophoblast layer into giant cells. We investigated if Ezh2 controls the fate of embryos at an earlier stage by treating with cycloheximide (CHX) or microinjecting short interfering RNA (siRNA) to restrict embryonic Ezh2 expression during preimplantation. CHX inhibited de novo EZH2 protein synthesis in zygotes, suggesting that EZH2 requires de novo synthesis during post-fertilisation stages. We found that loss of Ezh2 at the pronuclear stage caused severe growth retardation and reduced blastocyst formation. Expression of the pluripotency-associated markers Oct4, Sox2 and Nanog were significantly decreased in embryos that had been injected with Ezh2 siRNA. In addition, Ezh2 loss induced upregulated expression of genes related to the differentiation of germ layers, including Gata6, Hoxb1 and Hand1. Finally, apoptosis was increased in the blastocyst embryos with Ezh2 knockdown. Modification of histone H3-Lysine 27 de-methylation and tri-methylation (H3K27me2/3) was strongly reduced in Ezh2 siRNA embryos. We conclude that Ezh2 is essential for early preimplantation embryo development through the regulation of epigenetic modification and apoptosis.