Increased CO2 Levels during the First Half of Incubation at High Altitude Modifies Embryonic Development of Fertile Leghorn Breeder Eggs

The exchange of oxygen (O2) and carbon dioxide (CO2) within an incubator has a significant impact on embryonic development (ED) and hatching processes. This study examines the influence of non-ventilation (NV) conditions during the first ten days of incubation at high altitudes on Leghorn hens hatching eggs. Five hundred four hatching eggs were equally divided into three treatment groups and placed in twelve incubators (R = 4). The first group was subjected to standard ventilated conditions (V) during the setting phase. The ventilation inlet holes of the remaining incubators in the NV treatments were closed with either micropore (M) or polypropylene (P) tape, referred to as NVM and NVP groups, respectively. These two different airtight settings were intended to allow for a gradual rise in CO2 naturally generated by the embryos. Results indicate that carbon dioxide concentration gradually increased during the first half of incubation, reaching 1.42% in the NVM group and 1.20% in the NVP group, while the V condition group remained at 0.15%. From 10 days of incubation onwards, normal V conditions were restored in all incubators. The highest hatchability of fertile eggs (HFE) was shown by the NVP group (55.7%), followed by the V (52.6%) and NVM (38.6%) groups. The NVP group showed a greater yolk-free body mass (YFBM) from 10 days of incubation until the hatch basket transfer. NV conditions during the first 10 days of incubation at high altitude produced higher YFBM with gradually decreasing yolk sac mass. In comparison to the NVM and V conditions, the particular NVP condition showed a beneficial impact on the quality of hatched chicks. Sustaining NVP condition (1.2% of CO2) throughout the first half of incubation at high altitude generated the optimal environment in the incubator ensuring the best hatchability results. This study highlights how important it is for hatchery managers to recognize the influence of low O2 and high levels of CO2 on the development trajectories of Leghorn embryos during early incubation at high altitudes.

Population genomic analysis reveals key genetic variations and the driving force for embryonic callus induction capability in maize

Genetic transformation has been an effective technology for improving the agronomic traits of maize.However,it is highly reliant on the use of embryonic callus(EC)and shows a serious genotype dependence.In this study,we performed genomic sequencing for 80 core maize germplasms and constructed a high-density genomic variation map using our newly developed pipeline(MQ2Gpipe).Based on the induction rate of EC(REC),these inbred lines were categorized into three subpopulations.The low-REC germplasms displayed more abundant genetic diversity than the high-REC germplasms.By integrating a genome-wide selective signature screen and region-based association analysis,we revealed 95.23 Mb of selective regions and 43 REC-associated variants.These variants had phenotypic variance explained values ranging between 21.46 and 49.46%.In total,103 candidate genes were identified within the linkage disequilibrium regions of these REC-associated loci.These genes mainly participate in regulation of the cell cycle,regulation of cytokinesis,and other functions,among which MYB15 and EMB2745 were located within the previously reported QTL for EC induction.Numerous leaf area-associated variants with large effects were closely linked to several REC-related loci,implying a potential synergistic selection of REC and leaf size during modern maize breeding.

Early embryonic failure caused by a novel mutation in the TUBB8 gene:A case report

BACKGROUND This study aimed to explore the relationship between gene mutations and early embryonic development arrest and to provide more possibilities for the diagnosis and treatment of repeated implantation failure.CASE SUMMARY Here,we collected and described the clinical data of a patient with early embryonic development stagnation after repeated in vitro fertilization attempts for primary infertility at the Department Reproductive Center of Zaozhuang Maternal and Child Healthcare Hospital.We also detected the whole-exon gene of the patient's spouse and parents,and conducted bioinformatics analysis to determine the pathogenesis of the gene.CONCLUSION A novel mutant of the TUBB8 gene[c.602G>T(p.C201F)]was identified,and this mutant provided new data on the genotype-phenotype relationships of related diseases.

Cardiac differentiation is modulated by anti-apoptotic signals in murine embryonic stem cells

BACKGROUND Embryonic stem cells(ESCs)serve as a crucial ex vivo model,representing epiblast cells derived from the inner cell mass of blastocyst-stage embryos.ESCs exhibit a unique combination of self-renewal potency,unlimited proliferation,and pluripotency.The latter is evident by the ability of the isolated cells to differ-entiate spontaneously into multiple cell lineages,representing the three primary embryonic germ layers.Multiple regulatory networks guide ESCs,directing their self-renewal and lineage-specific differentiation.Apoptosis,or programmed cell death,emerges as a key event involved in sculpting and forming various organs and structures ensuring proper embryonic development.How-ever,the molecular mechanisms underlying the dynamic interplay between diffe-rentiation and apoptosis remain poorly understood.AIM To investigate the regulatory impact of apoptosis on the early differentiation of ESCs into cardiac cells,using mouse ESC(mESC)models-mESC-B-cell lym-phoma 2(BCL-2),mESC-PIM-2,and mESC-metallothionein-1(MET-1)-which overexpress the anti-apoptotic genes Bcl-2,Pim-2,and Met-1,respectively.METHODS mESC-T2(wild-type),mESC-BCL-2,mESC-PIM-2,and mESC-MET-1 have been used to assess the effect of potentiated apoptotic signals on cardiac differentiation.The hanging drop method was adopted to generate embryoid bodies(EBs)and induce terminal differentiation of mESCs.The size of the generated EBs was measured in each condition compared to the wild type.At the functional level,the percentage of cardiac differentiation was measured by calculating the number of beating cardiomyocytes in the manipulated mESCs compared to the control.At the molecular level,quantitative reverse transcription-polymerase chain reaction was used to assess the mRNA expression of three cardiac markers:Troponin T,GATA4,and NKX2.5.Additionally,troponin T protein expression was evaluated through immunofluorescence and western blot assays.RESULTS Our findings showed that the upregulation of Bcl-2,Pim-2,and Met-1 genes led to a reduction in the size of the EBs derived from the manipulated mESCs,in comparison with their wild-type counterpart.Additionally,a decrease in the count of beating cardiomyocytes among differentiated cells was observed.Furthermore,the mRNA expression of three cardiac markers-troponin T,GATA4,and NKX2.5-was diminished in mESCs overexpressing the three anti-apoptotic genes compared to the control cell line.Moreover,the overexpression of the anti-apoptotic genes resulted in a reduction in troponin T protein expression.CONCLUSION Our findings revealed that the upregulation of Bcl-2,Pim-2,and Met-1 genes altered cardiac differentiation,providing insight into the intricate interplay between apoptosis and ESC fate determination.

Maternal zinc alleviates tert-butyl hydroperoxide-induced mitochondrial oxidative stress on embryonic development involving the activation of Nrf2/PGC-1αpathway

Background Mitochondrial dysfunction induced by excessive mitochondrial reactive oxygen species(ROS)damages embryonic development and leads to growth arrest.Objective The purpose of this study is to elucidate whether maternal zinc(Zn)exert protective effect on oxidative stress targeting mitochondrial function using an avian model.Result In ovo injected tert-butyl hydroperoxide(BHP)increases(P<0.05)hepatic mitochondrial ROS,malondialdehyde(MDA)and 8-hydroxy-2-deoxyguanosine(8-OHdG),and decreases(P<0.05)mitochondrial membrane potential(MMP),mitochondrial DNA(mtDNA)copy number and adenosine triphosphate(ATP)content,contributing to mitochondrial dysfunction.In vivo and in vitro studies revealed that Zn addition enhances(P<0.05)ATP synthesis and metallothionein 4(MT4)content and expression as well as alleviates(P<0.05)the BHP-induced mitochondrial ROS generation,oxidative damage and dysfunction,exerting a protective effect on mitochondrial function by enhancing antioxidant capacity and upregulating the mRNA and protein expressions of Nrf2 and PGC-1α.Conclusions The present study provides a new way to protect offspring against oxidative damage by maternal Zn supplementation through the process of targeting mitochondria involving the activation of Nrf2/PGC-1αsignaling.

MicroRNA-146a Promotes Embryonic Stem Cell Differentiation towards Vascular Smooth Muscle Cells through Regulation of Kruppel-like Factor 4

Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.

Effects of LPA on the development of sheep in vitro fertilized embryos and attempt to establish sheep embryonic stem cells

Lysophosphatidic acid(LPA)is a small molecule glycerophospholipid,which regulates multiple downstream signalling pathways through G-protein-coupled receptors to achieve numerous functions on oocyte maturation and embryo development.In this study,sheep in vitro fertilized embryos were applied to investigate the effects of LPA on early embryos development and embryonic stem cell establishment.At first,the maturation medium containing estrus female sheep serum and synthetic oviduct fluid(SOF)were optimized for sheep IVF,and then the effects of LPA were investigated.From 0.1 to 10μmol L^(–1),LPA had no significant effect on the cleavage rate(P>0.05),but the maturation rate and blastocyst rate increased dependently with LPA concentration(P<0.05),and the blastocyst morphology was normal.When the LPA concentration was 15μmol L^(–1),the maturation rate,cleavage rate and blastocyst rate decreased significantly(P<0.05),and the blastocyst exhibited abnormal morphology and could not develop into highquality blastocyst.Besides,the exogenous LPA increases the expression of LPAR2,LPAR4,TE-related gene CDX-2and pluripotency-related gene OCT-4 in sheep early IVF embryos with the raise of LPA concentration from 0.1 to 10μmol L^(–1).The expression of LPAR2,LPAR4,CDX-2 and OCT-4 from the LPA-0.1μmol L^(–1)to LPA-10μmol L^(–1)groups in early embryos were extremely significant(P<0.05),while the expression of these genes significantly decreased in 15μmol L^(–1)LPA-treated embryos compared with LPA-10μmol L^(–1)group(P<0.05).The inner cell mass in 15μmol L^(–1)LPA-treated embryos was also disturbed,and the blastocysts formation was abnormal.Secondly,the sheep IVF blastocysts were applied to establish embryonic stem cells.The results showed that LPA made the blastocyst inoculated cells grow towards TSC-like cells.They enhanced the fluorescence intensity and mRNA abundance of OCT-4 and CDX-2 as the concentration increased from 0 to 10μmol L^(–1),while 15μmol L^(–1)LPA decreased OCT-4 and CDX-2 expression in the derived cells.The expression of CDX-2 and OCT-4 in the blastocyst inoculated cells of LPA-1μmol L^(–1)group and LPA-10μmol L^(–1)group extremely significantly increased(P<0.05),but there was significant decrease in LPA-15μmol L^(–1)group compared with LPA-10μmol L^(–1)group(P<0.05).Meanwhile,the protein expression of LPAR2 and LPAR4 remarkably increased after treatment of LPA at 10μmol L^(–1)concentration.This study references the IVF embryo production and embryonic stem cell research of domestic animals.

Effects of dietary coenzyme Q10 supplementation during gestation on the embryonic survival and reproductive performance of high‑parity sows

Background Fertility declines in high-parity sows.This study investigated whether parity-dependent declines in embryonic survival and reproductive performance could be restored by dietary coenzyme Q10(CoQ10)supplementation.Methods Two experiments were performed.In Exp.1,30 young sows that had completed their 2nd parity and 30 high-parity sows that had completed their 10^(th)parity,were fed either a control diet(CON)or a CON diet supple-mented with 1 g/kg CoQ10(+CoQ10)from mating until slaughter at day 28 of gestation.In Exp.2,a total of 314 post-weaning sows with two to nine parities were fed the CON or+CoQ10 diets from mating throughout gestation.Results In Exp.1,both young and high-parity sows had a similar number of corpora lutea,but high-parity sows had lower plasma CoQ10 concentrations,down-regulated genes involved with de novo CoQ10 synthesis in the endome-trium tissues,and greater levels of oxidative stress markers in plasma and endometrium tissues.High-parity sows had fewer total embryos and alive embryos,lower embryonic survival,and greater embryo mortality than young sows.Dietary CoQ10 supplementation increased the number of live embryos and the embryonic survival rate to levels simi-lar to those of young sows,as well as lowering the levels of oxidative stress markers.In Exp.2,sows showed a parity-dependent decline in plasma CoQ10 levels,and sows with more than four parities showed a progressive decline in the number of total births,live births,and piglets born effective.Dietary supplementation with CoQ10 increased the number of total births,live births,and born effective,and decreased the intra-litter covariation coefficients and the percentage of sows requiring farrowing assistance during parturition.Conclusions Dietary CoQ10 supplementation can improve the embryonic survival and reproductive performance of gestating sows with high parity,probably by improving the development of uterine function.

Whole‑genome transcriptome and DNA methylation dynamics of pre‑implantation embryos reveal progression of embryonic genome activation in buffaloes

Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related to the embryonic genome activation(EGA).During MZT,the embryos are sensitive to the environment and easy to arrest at this stage in vitro.However,the timing and regulation mechanism of EGA in buffaloes remain obscure.Results Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing(WGBS)to draw landscapes of transcription and DNA-methylation.Four typical developmental steps were classified during buffalo PED.Buffalo major EGA was identified at the 16-cell stage by the comprehensive analy-sis of gene expression and DNA methylation dynamics.By weighted gene co-expression network analysis,stage-spe-cific modules were identified during buffalo maternal-to-zygotic transition,and key signaling pathways and biological process events were further revealed.Programmed and continuous activation of these pathways was necessary for success of buffalo EGA.In addition,the hub gene,CDK1,was identified to play a critical role in buffalo EGA.Conclusions Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT.It will lay a foundation for improving the in vitro development of buffalo embryos.

Embryonic Development and Eclosion Season of New Species Berastagia (Lepidoptera: Pyralidae) from Taiwan

This paper describes a new species of the snout moth Berastagia tainanica sp. nov. (Lepidoptera: Pyralidae) from Taiwan. From 2009 to 2016, a biology study was conducted on population dynamics and embryonic development. Spring season is the peak of the eclosion of overwintering larvae or pupae. The average longevity of adult was 14.8 ± 6.2 days (N = 174), the average number of eggs laid was 259 ± 3 eggs/moth (N = 2), the hatching rate of eggs was 95.4% (N = 262), and the average hatching time of eggs was 99.6 ± 18.6 hours (N = 68). The average body length of males was 5.64 mm ± 0.91 mm (N = 30), and the average body length of females was 6.28 mm ± 0.84 mm (N = 30). This finding indicates that female snout moths are larger than males (Global R = 0.058, P = 0.012). The snout moth eclosion rate was 16.9 moths/100 pods in the first year (2010/2011, N = 2,224 pods) and 10.9 moths/100 pods in the second year (2014/2015, N = 6,382 pods). The pod borer rate was 31.8% (N = 707) and the seed borer rate was 41.2% (N = 3,628) in the first year, whereas the pod borer rate was 76.2% (N = 6,382) in the second year.

Visible Light and Its Influence on the Embryonic Viability of the Cricket Acheta domesticus

During in vitro fertilization, human embryos are incubated without light, and these conditions do not ensure embryo survival. This study explored whether environmental conditions can influence the embryo viability rates of the house cricket, Acheta domesticus. In particular, the experiment tested what colors of visible light provide the best incubation conditions to ensure cricket embryo viability. The concept was to use house cricket embryos to represent human embryos. Cricket embryos were chosen as their eggs have soft outer membrane casings and resemble human embryos during the first few days after fertilization. During the experiment, the adult crickets laid their eggs into one of six soil-filled boxes called substrates. Each substrate was placed into one of six storage containers filled with adult crickets and lit with a different colored visible light (red, yellow, green, blue, white, or no light). After two days of breeding, the egg-filled substrates were removed from the adult crickets and placed in another storage container of the same color light. After incubation under heat-emitting lamps and under one of six light colors, nymphs were counted after hatching to determine embryo viability. After three trials, the red light provided the significantly highest viability rate, with yellow and no light being comparable seconds. The green, blue, and white lights showed significantly lower viability rates than no visible light. My results raise the speculation that exposing fertilized mammal eggs to visible light colors might have the same effects during the in vitro fertilization process.

Embryonic, genetic and clinical outcomes of fresh versus vitrified oocyte: A retrospective cohort study

Objective:To compare embryonic development,ploidy status and clinical outcomes between fresh and frozen-thawed oocytes.Methods:This retrospective cohort study evaluated 83 fertilization cycles including both fresh and frozen oocytes from 79 patients at the HP Fertility Center of Hai Phong International Hospital of Obstetrics and Pediatrics in Vietnam.The patient underwent several ovarian stimulation cycles to accumulate a certain number of oocytes that would be vitrified.In the last oocyte retrieval,all patient’s oocytes including both frozen and fresh would be fertilized.The outcomes included the rates of oocyte survival,cleavage embryo,blastocyst,ploidy status,pregnancy,biochemical pregnancy and clinical pregnancy.Results:The oocyte survival rate after thawing was 96.5%.No statistically significant difference was found when comparing fresh and frozen oocytes regarding fertilization rate(78.1%vs.75.5%,P=0.461),usable cleavage embryo rate(86.9%vs.87.2%,P=0.916)but usable blastocyst rate was found higher statistically in the frozen oocyte group(44.4%vs.54.0%,P=0.049).The percentages of euploid,aneuploid and mosaic embryos between the fresh group and the vitrified group had no significant differences(33.8%vs.31.6%,P=0.682;51.0%vs.54.2%,P=0.569;15.2%vs.12.4%,P=0.787;respectively).The rates of pregnancy,biochemical pregnancy and clinical pregnancy had no statistical difference(68.8%vs.64.8%,P=0.764;12.5%vs.3.6%,P=0.258;37.5%vs.46.4%,P=0.565).17 Mature oocytes are the minimum to have at least one euploid embryo.Conclusions:Oocyte vitrification does not affect embryonic,genetic and clinical results.The number of mature oocytes should be considered for fertilization in some cases.

Endometrial Status and Embryonic Implantation about 150 Cases at the Medically Assisted Procreation Laboratory “Le Diafounou”

Background: The endometrium is the tissue that lines the inside of the uterus. It undergoes multiple changes during the menstrual cycle and prepares to welcome the fertilized egg. Endovaginal ultrasound is a first-line examination in the assessment of female infertility. Objective: We aimed to determine the impact of endometrial status by endovaginal ultrasound on the outcome of embryo transfers. Subjects and Methods: This was a prospective cross-sectional study of 150 women collected between January and October 2020 at the “Le Diafounou” medically assisted procreation laboratory. Were included in the study, women presenting for desire of pregnancy and having continued the treatment until the transfer of fresh or frozen embryos. An uterine score was used. This score took into account the thickness of endometrium, it is coffee bean aspect, the presence of the notch, the echogenicity of the endometrium, the pulsatility index, the presence of the flow at the end of diastole and sub-endometrial flow. Ultrasounds were performed using a General Electric logic 500, logic 9 and Voluson E8 device. The data was entered into Excel and then analyzed using SPSS version 21. Results: 150 women were selected for the study. 83 women or 55.3% were between 17 and 35 years old and 44.7% were over 35 years old with an upper limit of 50 years old. The number of women who became pregnant was 69% or 46%. The average thickness of the endometrium was 11.40 mm with extremes of 4.65 mm and 18.6 mm. There was a correlation between the thickness of the endometrium and obtaining a pregnancy (p i.e. 88.7%. The pulsatility index was relatively low for those who started a pregnancy, i.e. 90%. The high pulsatility index is one of the reasons for the failure of embryo implantation. The correlation was significant (p = 0.009). 141 women had no notch in the uterine arteries, i.e. 94%. 94 women or 62.7% had a triple line endometrium or typical coffee bean appearance. End-diastolic flow was observed in 127 women, i.e. 84.7%. The sub-endometrial flow was found in 128 women or 85.3%. Conclusion: The Knowledge of the status of the endometrium is essential and has an impact on the outcome of embryo transfers. The thinner (7 and <15). Doppler factors also play an important role.

Clinical Analysis of Embryonic Posterior Cerebral Artery

Objective:To understand the clinical characteristics of patients with embryonic posterior cerebral artery and its correlation with abnormal vascular development.Methods:The clinical data of 396 patients with embryonic posterior cerebral artery confirmed by magnetic resonance angiography(MRA)and computed tomography angiography(CTA)were analyzed.Results:Two-hundred patients had clinical manifestations of posterior circulation ischemia,including recurrent dizziness,vertigo,and tinnitus;45 had headaches,97 had limb weakness,and 16 patients had syncope or impaired consciousness.Seventy-six patients with circulatory infarction were admitted to the hospital.There were 251 patients with history of hypertension,74 with diabetes,113 with hyperlipidemia,13 with dominant vertebral artery,10 with intracranial aneurysm,and 19 with absence of A1 segment of the anterior cerebral artery(considering developmental variation).Conclusion:Embryonic posterior cerebral artery develops abnormally during the embryonic period,often accompanied by abnormal vascular access.Due to abnormal hemodynamics,the incidence of posterior circulation ischemia,aneurysm,and infarction increases in such patients.

脂滴功能及其与卵母细胞成熟和胚胎发育的关系研究进展

脂滴不仅是真核细胞内贮存甘油三酯和胆固醇酯等中性脂质的重要细胞器,也是细胞代谢和细胞器质量控制的中心调控者,参与细胞能量代谢、类固醇激素合成、内质网应激、线粒体功能以及生物膜结构形成等过程的调控。目前认为脂滴合成于内质网,并以出芽的方式进入细胞质。脂滴是呈动态分布的细胞器,其数量、大小以及分布受细胞获得营养物质和代谢状态影响。众所周知,哺乳动物卵母细胞含有脂滴,且卵母细胞中含有足够数量的脂滴与卵母细胞成熟和早期胚胎发育密切相关。更重要的是,最近的研究表明,脂滴在胚胎附植着床过程中发挥了至关重要的作用,即使卵母细胞中脂滴含量较低的哺乳动物,维持适量的脂滴对早期胚胎发育也很重要。因此,本文综述了脂滴的合成与分布,分析了脂滴的主要功能,重点阐述脂滴在动物卵母细胞成熟和早期胚胎发育期间的关键作用,并讨论脂滴卵母细胞和胚胎内脂滴含量与胚胎滞育持续时间的关系,以期为通过调控脂滴代谢而增强卵母细胞质量和胚胎发育能力,进而提高动物繁殖性能提供理论基础。

异丙隆原药对大鼠的致畸作用

目的:研究异丙隆原药对大鼠的致畸作用。方法:将受孕的雌性Wistar大鼠随机分为对照组(花生油)和低(36.9 mg/kg)、中(73.8 mg/kg)、高(369.0 mg/kg)3个异丙隆原药染毒剂量组,每组15只孕鼠。对照组和各染毒组于大鼠妊娠第6~15天灌胃给予受试物,每天1次,连续10 d。于妊娠第20天处死各组孕鼠,观察母鼠和胎鼠的生长、发育情况。结果:与对照组比较,低、中剂量组孕鼠各项观察指标差异均无统计学意义(P>0.05)。高剂量组孕鼠于受孕第9天出现萎靡不振、背毛蓬松、体质量下降(P<0.01);着床数、活胎数减少,死胎数增多,窝均体质量降低(P<0.5);部分胎鼠出现囟门大、枕骨及胸骨骨化不全、肋骨弯曲,骨骼畸形率为27.78%,与对照组相比,差异均具有统计学意义(P<0.05)。结论:在本实验条件下,369.0 mg/kg组异丙隆原药可引起Wistar大鼠的母体毒性和胚胎发育毒性。

形态动力学联合减数分裂纺锤体观测在卵裂期胚胎筛选中的应用

目的利用胚胎形态动力学联合卵母细胞减数分裂纺锤体(MS)观测,探索预测胚胎发育潜能的卵裂期胚胎筛选方法。方法回顾性分析2020年8月至2022年6月在苏州市立医院生殖与遗传中心行卵胞浆内单精子注射(ICSI),同时行卵母细胞MS观测和胚胎实时观测培养的46个周期患者的临床资料。根据卵母细胞MS形态、胚胎是否存在异常分裂方式,以及胚胎形态学差异对卵母细胞及卵裂期胚胎进行分类研究,分析胚胎形态动力学参数及MS形态预测胚胎发育潜能的价值,建立预测囊胚形成的早期胚胎分层筛选模型,并利用受试者工作特征(ROC)曲线探讨该模型的预测价值。结果MS正常组的囊胚形成率显著高于MS异常组(63.6%vs.46.9%,P<0.05);直接分裂(DC)1组的囊胚形成率显著低于DC2组和无DC组(5.0%vs.46.7%、63.9%,P<0.05);D3形态学优质胚胎的囊胚形成率显著高于D3形态学非优质胚胎(79.4%vs.24.6%,P<0.05)。采用多元Logistic回归分析发现,在D3形态学优质胚胎中,3细胞到5细胞的发育时间(cc3)、5细胞到8细胞的发育时间(s3)和MS形态对囊胚形成有预测价值(P<0.05);在D3形态学非优质胚胎中,卵裂球数目对囊胚形成有预测价值(P<0.05)。以此建立了D3胚胎分层筛选模型,共11级(A~K级),A级最优,K级最劣。该筛选模型对囊胚形成的ROC曲线显示,曲线下面积为0.857[95%CI(0.816,0.899),P=0.000]。结论采用胚胎形态动力学参数联合MS形态分析建立的D3胚胎分层筛选模型,对胚胎发育潜能具有较好的预测能力。

儿童肝脏未分化胚胎性肉瘤的影像表现及病理分析

目的:分析儿童肝脏未分化胚胎性肉瘤(undifferentiated embryonal sarcoma of the liver,UESL)的超声、CT及MRI影像学表现,以提高对该病的认识水平。方法:回顾性收集2009年12月至2021年12月首都儿科研究所附属儿童医院经手术及病理证实的11例儿童UESL,分析所有患儿的超声、CT及MRI影像表现,总结其影像学特点。结果:病灶均为单发,肿块最大径11.5~19.8 cm。CT平扫显示所有病灶边缘较清楚、其内呈不均匀低密度,内可见厚薄不均的分隔;囊性密度区边缘或分隔周围可见少许片絮样软组织密度影。6例超声图像均提示实性占位,表现为混合回声的实性包块内可见大小不等的无回声区。3例MRI表现为边界清楚肿块影,T1WI呈不均匀混杂低信号,病灶内可见高信号区;T2WI呈混合高信号区和条状低信号区。9例病灶增强扫描动脉期均以边缘强化为主,8例可见迂曲增粗血管影;延迟期病灶呈持续不均匀强化,边缘持续强化,且从外围向中心充盈,强化范围增大,8例延迟期可见假包膜。结论:儿童UESL影像学表现有一定特征性,了解其影像特点并结合临床特征,有助于早期诊断。

红鲫胚胎发育过程中Sox8基因对壬基酚胁迫的响应

【目的】明确红鲫SRY相关高迁移率族盒蛋白8基因(Sox8)表达模式及壬基酚(NP)暴露对红鲫胚胎发育过程中Sox8基因表达的影响,为揭示NP胁迫下红鲫胚胎发育畸形的分子机制提供科学依据。【方法】运用RACE克隆红鲫Sox8基因cDNA全长序列,通过ProtParam、ProtScale、InterPro、PredictProtein、DeepTMHMM等在线软件进行生物信息学分析,利用实时荧光定量PCR检测红鲫Sox8基因在各成体组织及不同胚胎发育阶段的表达情况,以及不同浓度NP暴露对红鲫胚胎期Sox8基因表达的影响。【结果】红鲫Sox8基因c DNA序列全长2163 bp,其开放阅读框(ORF)1176bp,共编码391个氨基酸残基;红鲫SOX8蛋白相对分子量为43798.30Da,理论等电点(pI)为6.90,脂溶指数为49.92,不稳定指数为66.04,总平均亲水性指数(GRAVY)为-1.026,其N端含有Sox_N二聚化结构域和HMG-box结构域;基于SOX8氨基酸序列相似性构建的系统发育进化树显示红鲫与鲫的亲缘关系最近。红鲫Sox8基因在各成体组织中广泛表达,以脑组织中的Sox8基因相对表达量最高,显著高于其他组织(P<0.05,下同);在红鲫胚胎发育过程中,Sox8基因的表达从受精后到14体节期呈逐渐下降趋势,但从21体节期开始Sox8基因相对表达量显著增高,直到孵化期始终维持在较高水平。在大多数情况下,NP暴露能显著影响红鲫胚胎发育过程中Sox8基因的表达,特别是14体节期后的发育阶段,NP暴露主要是不同程度地激活Sox8基因表达。【结论】Sox8基因表达在红鲫成体中具有组织特异性,且在胚胎发育过程中呈先降低后升高的变化趋势。NP暴露能显著影响红鲫胚胎发育过程中Sox8基因的表达,14体节期后Sox8基因表达水平升高可能是NP致畸红鲫胚胎的重要原因之一。

术前血清CEA、CA153、CA724及CA199水平与直肠癌临床分期及预后的关系

目的探讨术前血清癌胚抗原(CEA)、糖类抗原153(CA153)、糖类抗原724(CA724)及糖类抗原199(CA199)水平与直肠癌临床分期及预后的关系。方法随机选取196例直肠癌患者为研究对象,比较不同分期及不同预后患者术前血清CEA、CA153、CA724和CA199水平。采用多因素Logistic回归分析直肠癌患者预后不良的影响因素;采用受试者工作特征(ROC)曲线分析术前血清CEA、CA153、CA724及CA199水平对直肠癌患者预后的预测价值。结果Ⅰ~Ⅳ期直肠癌患者术前血清CEA、CA153、CA724及CA199水平差异具有统计学意义(P<0.05),且随着分期的增加呈现上升趋势。预后不良组和预后良好组在淋巴结转移、分化程度、浸润深度及肿瘤直径方面比较差异具有统计学意义(P<0.05);与预后良好组比较,预后不良组术前血清CEA、CA153、CA724及CA199水平均明显较高(P<0.05);多因素Logistic回归分析结果显示,直肠癌患者血清CEA、CA153、CA724、CA199、淋巴结转移、低分化程度、浸润深度(T3~T4)为预后不良的独立危险因素(P<0.05)。ROC结果显示,血清CEA、CA153、CA724及CA199水平单独预测直肠癌患者预后的AUC分别为0.825、0.803、0.793、0.766,灵敏度分别为73.08%、67.31%、69.23%、71.15%,特异度分别为85.61%、84.89%、86.33%、87.05%;血清CEA、CA153、CA724及CA199水平联合预测直肠癌患者预后的AUC为0.883(95%CI:0.828~0.925),显著大于血清CEA单独预测的AUC(Z=2.134,P=0.033)、CA153单独预测的AUC(Z=2.167,P=0.030)、CA724单独预测的AUC(Z=2.288,P=0.022)和CA199单独预测的AUC(Z=2.801,P=0.005)。结论直肠癌患者术前血清CEA、CA153、CA724和CA199水平与临床分期和预后有关,且四者联合检测对患者预后具有较高的预测价值。