Factors influencing procalcitonin in the cerebrospinal fluid of patients after neurosurgery and its diagnostic value for intracranial infection.

OBJECTIVE: This study aimed to investigate the factors influencing Procalcitonin (PCT) in the cerebrospinal fluid (CSF) of patients with high fever and suspected intracranial infection after neurosurgery and its clinical application value. METHODS: Between February 2021 and August 2022, CSF and serum samples were collected via lumbar puncture from patients with high fever and suspected intracranial infection in the Intensive Care Unit(ICU) of our hospital. Multivariate logistic regression analysis was performed to analyze the factors influencing elevated PCT in CSF. The diagnostic efficacy of each index was assessed using receiver operating characteristic (ROC) curves. RESULTS: A total of 183 CSF samples were collected, of which 148 had increased PCT levels, including 73 cases of intracranial infection and 75 cases in the case‒control group. Multivariate logistic regression analysis showed that intracranial infection [OR = 0.117, 95% CI: 0.025-0.559; p < 0.01] and hemorrhagic CSF [OR = 0.162, 95% CI: 0.029-0.916; p < 0.04] were factors influencing CSF PCT, while trauma [OR = 3.43, 95% CI: 0.76-15.45; p < 0.12], epileptic seizure [OR = 0.00, 95% CI: 0.00; p < 0], age [OR = 1.02, 95% CI: 0.98-1.52; p < 0.32] and Glasgow Coma Scale (GCS) score [OR = 1.03, 95% CI: 0.78-1.32; p < 0.83] did not influence CSF PCT. The CSF PCT and serum PCT levels in the intracranial infection group and the case‒control group were 0.13 (0.11, 0.25) ng/ml and 0.14 (0.07, 0.25) ng/ml and 0.14 (0.08,0.32) ng/ml and 0.23 (0.13,0.48)ng/ml, respectively, with no statistically significant difference. The median values of CSF lactate in the intracranial infection group and the case‒control group were 6.45 (4.475, 8.325) mmol/l and 3.2 (2.02, 4.200) mmol/l, respectively, with a statistically significant difference between the groups.The areas under the ROC curve of CSF PCT, serum PCT,CSF lactate, CSF PCT combined with lactate were 0.59, 0.63, 0.82,and 0.83,respectively. CONCLUSION: Intracranial infection and hemorrhagic CSF are influencing factors for elevated CSF PCT following neurosurgery. It should be noted that the diagnostic value of intracranial infection by CSF PCT elevated alone is limited, but the combination it with other indicators can help improve diagnostic efficacy.

The RNA m6A landscape of mouse oocytes and preimplantation embryos.

Despite the significance of N6-methyladenosine (m6A) in gene regulation, the requirement for large amounts of RNA has hindered m6A profiling in mammalian early embryos. Here we apply low-input methyl RNA immunoprecipitation and sequencing to map m6A in mouse oocytes and preimplantation embryos. We define the landscape of m6A during the maternal-to-zygotic transition, including stage-specifically expressed transcription factors essential for cell fate determination. Both the maternally inherited transcripts to be degraded post fertilization and the zygotically activated genes during zygotic genome activation are widely marked by m6A. In contrast to m6A-marked zygotic ally-activated genes, m6A-marked maternally inherited transcripts have a higher tendency to be targeted by microRNAs. Moreover, RNAs derived from retrotransposons, such as MTA that is maternally expressed and MERVL that is transcriptionally activated at the two-cell stage, are largely marked by m6A. Our results provide a foundation for future studies exploring the regulatory roles of m6A in mammalian early embryonic development.

Novel mitochondrial tRNALeu(UUR) 3261A > g mutation in two pedigrees with essential hypertension.

BACKGROUND: Essential hypertension (EH) was associated with mitochondrial tRNA mutations. AIMS: This study was designed to assess the association between EH and mitochondrial dysfunction. METHODS: A total of 30 individuals from two different Chinese families exhibit maternally inherited EH were assessed for genetic, clinical, and biochemical phenotypes pertaining to EH and mitochondrial functionality. These analyses included assessments of tRNALeu(UUR) 3261A > G mutation status, mitochondrial membrane permeability, mitochondria-associated ATP and reactive oxygen species (ROS) generation, and electron transport chain functionality. RESULTS: EH was detected in 6 total analyzed members of the two families assessed in the present study, with its initial age of onset and presentation varying among patients. These patients with EH exhibited the tRNALeu(UUR) 3261A > G mutation and were of the B5 and D4 Eastern Asian mitochondrial haplogroups. This 3261A > G mutation was predicted to result in disruption of normal tRNALeu(UUR) activity owing to the destabilization of conserved base pairing (30A-40U). Consistent with this prediction, we found that cybrid cell lines exhibiting this 3261A > G mutation exhibited a ~49.05% decrease in baseline tRNALeu(UUR) levels. These cells additionally exhibited ~44.81% reductions in rates of mitochondrial translation. CONCLUSIONS: To facilitate future molecular diagnosis, the 3261A > G mutation should be included in the list of hereditary risk factors. Our findings will aid in the counseling of EH families.

CENP-F-dependent DRP1 function regulates APC/C activity during oocyte meiosis I.

Chromosome segregation is initiated by cohesin degradation, which is driven by anaphase-promoting complex/cyclosome (APC/C). Chromosome cohesin is removed by activated separase, with the degradation of securin and cyclinB1. Dynamin-related protein 1 (DRP1), a component of the mitochondrial fission machinery, is related to cyclin dynamics in mitosis progression. Here, we show that DRP1 is recruited to the kinetochore by centromeric Centromere protein F (CENP-F) after nuclear envelope breakdown in mouse oocytes. Loss of DRP1 during prometaphase leads to premature cohesin degradation and chromosome segregation. Importantly, acute DRP1 depletion activates separase by initiating cyclinB1 and securin degradation during the metaphase-to-anaphase transition. Finally, we demonstrate that DRP1 is bound to APC2 to restrain the E3 ligase activity of APC/C. In conclusion, DRP1 is a CENP-F-dependent atypical spindle assembly checkpoint (SAC) protein that modulates metaphase-to-anaphase transition by controlling APC/C activity during meiosis I in oocytes.

Successful treatment of ceftazidime/avibactam combined with aztreonam in the NDM-producing Klebsiella pneumoniae bloodstream and intestinal infections in a NK/T lymphoma patient with agranulocytosis during autologous hematopoietic stem cell transplantation: a case report.

New Delhi metallo-beta-lactamase (NDM)-producing Klebsiella pneumoniae is increasingly reported worldwide. Clinicians face significant challenges in the treatment of this multidrug-resistant bacterium. The combination of ceftazidime/avibactam (CAZ/AVI) and aztreonam (ATM) is currently probably the most effective strategy for the treatment of such infection. We described a patient diagnosed with NK/T cell lymphoma who underwent autologous hematopoietic stem cell transplantation (ASCT) in the hematology department. The patient developed severe infection after ASCT. Blood and stool cultures showed carbapenem-resistant K. pneumoniae. Blood sample was detected as NDM-producing K. pneumoniae. We successfully treated this infection with CAZ/AVI and ATM.

Hexagonal Network of Photocurrent Enhancement in Few-Layer Graphene/InGaN Quantum Dot Junctions.

Strain in two-dimensional (2D) materials has attracted particular attention because of the remarkable modification of electronic and optical properties. However, emergent electromechanical phenomena and hidden mechanisms, such as strain-superlattice-induced topological states or flexoelectricity under strain gradient, remain under debate. Here, using scanning photocurrent microscopy, we observe significant photocurrent enhancement in hybrid vertical junction devices made of strained few-layer graphene and InGaN quantum dots. Optoelectronic response and photoluminescence measurements demonstrate a possible mechanism closely tied to the flexoelectric effect in few-layer graphene, where the strain can induce a lateral built-in electric field and assist the separation of electron-hole pairs. Photocurrent mapping reveals an unprecedentedly ordered hexagonal network, suggesting the potential to create a superlattice by strain engineering. Our work provides insights into optoelectronic phenomena in the presence of strain and paves the way for practical applications associated with strained 2D materials.

Analysis of Prognostic Risk Factors of Sepsis Patients in Intensive Care Unit Based on Data Analysis.

In this paper, a data-enabled analysis of the prognostic risk factors of sepsis patients in the intensive care unit is presented. For this purpose, we have selected 220 sepsis patients, preferably those admitted to the intensive care unit for treatment in a tertiary a hospital in Tianjin from June 2018 to June 2019 and received complete data as the research objects, to explore the prognostic risk factors of sepsis patients in the intensive care unit. All patients met the SSC sepsis diagnosis guidelines and recorded the patients' age, gender, underlying disease, and infection site. Laboratory indicators, such as blood routine, electrolytes, arterial blood gas, liver function, and renal function, were collected within 24 hours of admission. Furthermore, the corresponding specimens were cultured for pathogenic microorganisms according to the site of infection. The LAC value was measured at admission and 24 h after admission, and the 24 h lactate clearance rate was calculated. The Acute Physiological and Chronic Health Status Score II (APACHE-II) and SOFA score were calculated, which were based on the worst value of the index within 24 hours after admission. According to the prognosis of patients during hospitalization, they are divided into two groups: (i) survival group and (ii) death group. We entered all the data into Excel and used SPSS21.0 statistical software for data analysis and processing. Quantitative data are tested for normality. Quantitative data for normal distribution are expressed as mean ± standard deviation, and normal distribution and uniform variance are measured. The factors affecting the prognosis of patients with sepsis were first subjected to a single-factor logistic regression analysis, and a multiple logistic regression analysis was performed on the basis of the significance of the single-factor analysis. The results found that the prognosis of patients with sepsis in the ICU is affected by multiple factors such as underlying diseases, infectious microorganisms, comorbidities, and interventional therapy. APACHE-II score, 24 h lactate clearance rate, ARDS, and DIC are independent risk factors that affect the prognosis of ICU patients.

Saikosaponin-d Alleviates Renal Inflammation and Cell Apoptosis in a Mouse Model of Sepsis via TCF7/FOSL1/Matrix Metalloproteinase 9 Inhibition.

Evidence exists reporting that saikosaponin-d (Sa) can prevent experimental sepsis, and this study aims to illustrate the molecular events underlying its renoprotective effects on lipopolysaccharide (LPS)-induced renal inflammation simulating sepsis. Through network pharmacology analysis and bioinformatics analysis, we identified that Sa may influence sepsis development by mediating TCF7. Dual luciferase reporter gene and chromatin immunoprecipitation (ChIP) assays were used to explore the interactions between TCF7, FOSL1, and matrix metalloproteinase 9 (MMP9). The experimental data suggest that Sa attenuated LPS-induced renal injury, as evidenced by the reduced production of proinflammatory cytokines as well as cell apoptosis in the renal tissues of LPS-induced mice. Mechanically, Sa inhibited FOSL1 by inhibiting TCF7, which reduced the expression of inflammatory factors in renal cells. TCF7 activated the FOSL1 expression and consequently promoted the expression of MMP9. Also, Sa reduced cell apoptosis and the expression of inflammatory factors by inhibiting the TCF7/FOSL1/MMP9 axis in vivo. In conclusion, Sa suppresses FOSL1 transcription by downregulating TCF7, thereby inhibiting MMP9 expression and ultimately reducing the renal inflammation and cell apoptosis induced by sepsis.

Clathrin Heavy Chain 1 Plays Essential Roles During Oocyte Meiotic Spindle Formation and Early Embryonic Development in Sheep.

As a major protein of the polyhedral coat of coated pits and vesicles, clathrin molecules have been shown to play a stabilization role for kinetochore fibers of the mitotic spindle by acting as inter-microtubule bridges. Clathrin heavy chain 1 (CLTC), the basic subunit of the clathrin coat, plays vital roles in both spindle assembly and chromosome congression during somatic-cell mitosis. However, its function in oocyte meiotic maturation and early embryo development in mammals, especially in domesticated animals, has not been fully investigated. In this study, the expression profiles and functional roles of CLTC in sheep oocytes were investigated. Our results showed that the expression of CLTC was maintained at a high level from the germinal vesicle (GV) stage to metaphase II stage and that CLTC was distributed diffusely in the cytoplasm of cells at interphase, from the GV stage to the blastocyst stage. After GV breakdown (GVBD), CLTC co-localized with beta-tubulin during metaphase. Oocyte treatments with taxol, nocodazole, or cold did not affect CLTC expression levels but led to disorders of its distribution. Functional impairment of CLTC by specific morpholino injections in GV-stage oocytes led to disruptions in spindle assembly and chromosomal alignment, accompanied by impaired first polar body (PB1) emissions. In addition, knockdown of CLTC before parthenogenetic activation disrupted spindle formation and impaired early embryo development. Taken together, the results demonstrate that CLTC plays a vital role in sheep oocyte maturation via the regulation of spindle dynamics and an essential role during early embryo development.

MiR-455-5p monitors myotube morphogenesis by targeting mylip.

As a posttranscriptional regulatory factor, microRNA (miRNA) plays an important role in the formation of myotubes. However, little is known about the mechanism of miRNA regulating myotube morphogenesis. Here, we aimed to characterize the function of miR-455-5p in myotube morphogenesis by inducing differentiation in C2C12 myoblasts containing murine Mylip fragments with the miR-455-5p target sequence. We found that miR-455-5p overexpression promoted the differentiation and hypertrophy of myotubes, while miR-455-5p inhibition led to the failure of myotube differentiation and formation of short myotubes. Furthermore, we demonstrated that miR-455-5p directly targeted the Mylip 3'-untranslated region, which plays a key role in monitoring myotube morphogenesis. Interestingly, the expression and function of Mylip were opposite to those of miR-455-5p during myogenesis. Our data uncovered novel miR-455-5p targets and established a functional link between Mylip and myotube morphogenesis. Understanding the involvement of Mylip in myotube morphogenesis provides insight into the function of the gene regulatory network.

Phycocyanin Improves Reproductive Ability in Obese Female Mice by Restoring Ovary and Oocyte Quality.

Reproductive dysfunction associated with obesity is increasing among women of childbearing age. Emerging evidence indicates that maternal obesity impairs embryo development and offspring health, and these defects are linked to oxidative stress in the ovary and in oocytes. Phycocyanin (PC) is a biliprotein from Spirulina platensis that possesses antioxidant, anti-inflammatory, and radical-scavenging properties. Our previous studies have shown that PC can reduce reactive oxygen species (ROS) accumulation in oocytes in D-gal-induced aging mice. Here, at the Institute of Cancer Research (ICR) mice fed a high-fat diet (HFD) to model obesity were used to test the effect of PC on reversing the fertility decline caused by obesity. We observed a significant increase in litter size and offspring survival rates after PC administration to obese mice. Further, we found that PC not only ameliorated the level of ovarian antioxidant enzymes, but also reduced the occurrence of follicular atresia in obese female mice. In addition, the abnormal morphology of the spindle-chromosome complex (SCC), and the abnormal mitochondrial distribution pattern in oocytes both recovered. The obesity-related accumulation of ROS, increased number of early apoptotic cells, and the abnormal expression of H3K9me3 in oocytes were all partially reversed after PC administration. In summary, this is the first demonstration that PC can improve fertility by partially increasing ovarian and oocyte quality in obese female mice and provides a new strategy for clinically treating obesity-related infertility in females.

Oral administration of olaquindox negatively affects oocytes quality and reproductive ability in female mice.

As an effective feed additive in the livestock industry, olaquindox (OLA) has been widely used in domestic animal production. However, it is unclear whether OLA has negative effects on mammalian oocyte quality and fetal development. In this study, toxic effects of OLA were tested by intragastric gavage ICR mice with water, low-dose OLA (5 mg/kg/day), or high-dose OLA (60 mg/kg/day) for continuous 45 days. Results showed that high-dose OLA gavage severely affected the offspring birth and growth. Significantly, high-dose OLA impaired oocyte maturation and early embryo development, indicated by the decreased percentage of germinal vesicle breakdown, first polar body extrusion and blastocyst formation. Meanwhile, oxidative stress levels were increased in oocytes or ovaries, indexed by the increased levels of ROS, MDA, H2O2, NO, and decreased levels of GSH, SOD, CAT, GSH-Px and GSH-Rd. Furthermore, aberrant mitochondria distribution, defective spindle assembly, abnormal H3K4me2/H3K9me3 levels, increased DNA double-strand breaks and early apoptosis rate, were observed after high-dose OLA gavage. Taken together, our results for the first time illustrated that high-dose OLA gavage led to sub-fertility of females, which means that restricted utilization of OLA as feed additive should be considered.

Protein regulator of cytokinesis 1 regulates chromosome dynamics and cytoplasmic division during mouse oocyte meiotic maturation and early embryonic development.

In contrast to the homeokinesis of mitosis, asymmetric division of cytoplasm is the conspicuous feature of meiosis in mammalian oocytes. Protein regulator of cytokinesis 1 (PRC1) is an important regulator during mitotic spindle assembly and cytoplasmic division, but its functions in oocyte meiosis and early embryo development have not been fully elucidated. In this study, we detected PRC1 expression and localization and revealed a nuclear, spindle midzone-related dynamic pattern throughout meiotic and mitotic progressions. Treatment of oocytes with the reagents taxol or nocodazole disturbed the distribution of PRC1 in metaphase II oocytes. Further, PRC1 depletion led to failure of first polar body (PB1) extrusion and spindle migration, aneuploidy and defective kinetochore-microtubule attachment and spindle assembly. Overexpression of PRC1 resulted in PB1 extrusion failure, aneuploidy and serious defects of spindle assembly. To investigate PRC1 function in early embryos, we injected Prc1 morpholino into zygotes and 2-cell stage embryos. Depletion of PRC1 in zygotes impaired 4-cell, morula and blastocyst formation. Loss of PRC1 in single or double blastomeres in 2-cell stage embryos significantly impaired cell division, indicating its indispensable role in early embryo development. Co-immunoprecipitation showed that PRC1 interacts with polo-like kinase 1 (PLK1), and functional knockdown and rescue experiments demonstrated that PRC1 recruits PLK1 to the spindle midzone to regulate cytoplasmic division during meiosis. Finally, kinesin family member 4 knockdown downregulates PRC1 expression and leads to PRC1 localization failure. Taken together, our data suggest PRC1 plays an important role during oocyte maturation and early embryonic development by regulating chromosome dynamics and cytoplasmic division.

[Application of lung ultrasonography on evaluating extravascular lung water and prognosis in patients with acute respiratory distress syndrome].

OBJECTIVE: To evaluate the value of lung ultrasonography score (LUS) on assessing extravascular lung water (EVLW) and prognosis in patients with acute respiratory distress syndrome (ARDS). METHODS: The clinical data of 46 patients meeting ARDS Berlin definition admitted to intensive care unit (ICU) of Ningbo Yinzhou People's Hospital from July 2016 to December 2019 were retrospectively collected. The general data, vital signs, blood lactic acid (Lac), oxygenation index (OI), LUS, extravascular lung water index (EVLWI), sequential organ failure assessment (SOFA) score, clinical pulmonary infection score (CPIS) and the length of ICU stay were collected. According to the prognosis of patients during ICU treatment, the patients were divided into survival group and non-survival group, and the clinical characteristics between the two groups were compared. The correlation between LUS and OI, EVLWI, SOFA, and CPIS were analyzed by Pearson correlation analysis. Receiver operator characteristic (ROC) curve was plotted to determine the prognostic value of LUS for ARDS patients during ICU treatment. RESULTS: Forty-six patients were enrolled in the analysis, of whom 32 patients survived (69.6%), and 14 patients died (30.4%) during ICU treatment. There was no significant difference in gender, age, left ventricular ejection fraction (LVEF) or heart rate (HR) between the two groups. Compared with the survival group, the mean arterial pressure (MAP) and OI in the non-survival group were significantly lowered [MAP (mmHg, 1 mmHg = 0.133 kPa): 57.48±33.34 vs. 85.45±19.56, OI (mmHg): 74.50±18.40 vs. 233.06±28.28, both P < 0.05], while Lac, LUS, EVLWI, SOFA and CPIS were significantly increased [Lac (mmol/L): 6.78±2.56 vs. 2.21±1.42, LUS score: 23.57±2.03 vs. 15.58±2.24, EVLWI (mL/kg): 22.93±2.56 vs. 12.96±2.18, SOFA score: 20.21±3.35 vs. 12.43±2.97, CPIS score: 8.07±1.38 vs. 4.59±1.04, all P < 0.01], and the length of ICU stay was significantly shortened (days: 9.33±3.28 vs. 16.89±4.12, P < 0.05]. Pearson correlation analysis showed that a significant negative linear correlation was found between LUS and OI (r = -0.823, P < 0.01), and positive linear correlations were found between LUS and EVLWI, SOFA, CPIS (r values were 0.745, 0.614, 0.757, respectively, all P < 0.01). ROC curve analysis showed that both LUS and EVLWI could predict the prognosis of ARDS patients during ICU treatment, and the areas under ROC curve (AUC) of LUS and EVLWI were 0.936 and 0.991, respectively. When the cut-off of LUS score was 20.5, the sensitivity and specificity were 85.7% and 81.2% respectively. CONCLUSIONS: LUS score has a good correlation with EVLWI monitored by pulse index continuous cardiac output (PiCCO), which can reflect lung water content. LUS score can be used as an early prognostic indicator for ARDS patients.

[Methods used for gene manipulation in mammal female germ cells and early embryos].

For sexual reproduction, oocytes are mammalian female germ cells that provide the majority of maternal genetic material for early stage embryo production and development. Early stage embryos begin the process of multicellular organism formation through cell differentiation. Studies on mammalian female germ cells (oocytes) not only reveal its unique physiological characteristics, but also help understand the mechanism involved in cell differentiation of other cell types. However, because it is difficult to culture in vitro, our understanding of the function of oocytes and early stage embryos remains very limited. Gene editing or manipulation is one of the most commonly used method, which is also useful in the field of gametes study. In this review, we summarized the principles, advantages and disadvantages of techniques, which include conditional knockout, RNA interference, Morpholino, Trim-Away and antibody-mediated inhibition of protein function, currently used for gene manipulation in oocytes and early stage embryos. We also discuss the issues the investigators need to consider. Finally, we highlight the future directions for gene manipulation or editing in female germ cells and early stage embryos.

Blastocyst hatching site is regularly distributed and does not influence foetal development in mice.

Hatching out from the zona pellucida (ZP) is a crucial step for blastocyst implantation and development. However, it is still unknown whether the location of the hatching site relative to the inner cell mass (ICM) affects embryo implantation and foetal development. Here, we classified hatching blastocysts into three categories, 0° ≤ θ ≤ 30°, 30° < θ ≤ 60°, and 60° < θ ≤ 90°, in which θ is determined based on the relative position of the hatching site to the arc midpoint of the ICM. Non-surgical embryo transfer (NSET) devices were employed to evaluate blastocyst implantation and embryo development. Of 1,827 hatching blastocysts, 43.84%, 30.60%, and 21.67% were categorized as 30° < θ ≤ 60°, 0° ≤ θ ≤ 30°, and 60° < θ ≤ 90°, respectively. Embryos with different hatching sites showed no distinct differences in blastocyst implantation; surrogate female pregnancy; embryo development to term; litter size, or offspring survival, gender, or body weight. Our results indicate that mouse blastocyst hatching site is not randomly distributed. Embryo implantation and development are not correlated with the blastocyst hatching site in mice. Thus, assessment of the blastocyst hatching site should not be recommended to evaluate mouse blastocyst implantation and developmental potential.

Combined use of dbcAMP and IBMX minimizes the damage induced by a long-term artificial meiotic arrest in mouse germinal vesicle oocytes.

Phosphodiesterase (PDE)-mediated reduction of cyclic adenosine monophosphate (cAMP) activity can initiate germinal vesicle (GV) breakdown in mammalian oocytes. It is crucial to maintain oocytes at the GV stage for a long period to analyze meiotic resumption in vitro. Meiotic resumption can be reversibly inhibited in isolated oocytes by cAMP modulator forskolin, cAMP analog dibutyryl cAMP (dbcAMP), or PDE inhibitors, milrinone (Mil), Cilostazol (CLZ), and 3-isobutyl-1-methylxanthine (IBMX). However, these chemicals negatively affect oocyte development and maturation when used independently. Here, we used ICR mice to develop a model that could maintain GV-stage arrest with minimal toxic effects on subsequent oocyte and embryonic development. We identified optimal concentrations of forskolin, dbcAMP, Mil, CLZ, IBMX, and their combinations for inhibiting oocyte meiotic resumption. Adverse effects were assessed according to subsequent development potential, including meiotic resumption after washout, first polar body extrusion, early apoptosis, double-strand DNA breaks, mitochondrial distribution, adenosine triphosphate levels, and embryonic development. Incubation with a combination of 50.0 µM dbcAMP and 10.0 µM IBMX efficiently inhibited meiotic resumption in GV-stage oocytes, with low toxicity on subsequent oocyte maturation and embryonic development. This work proposes a novel method with reduced toxicity to effectively arrest and maintain mouse oocytes at the GV stage.

Loss of CENPF leads to developmental failure in mouse embryos.

Aneuploidy caused by abnormal chromosome segregation during early embryo development leads to embryonic death or congenital malformation. Centromere protein F (CENPF) is a member of centromere protein family that regulates chromosome segregation during mitosis. However, its necessity in early embryo development has not been fully investigated. In this study, expression and function of CENPF was investigated in mouse early embryogenesis. Detection of CENPF expression and localization revealed a cytoplasm, spindle and nuclear membrane related dynamic pattern throughout mitotic progression. Farnesyltransferase inhibitor (FTI) was employed to inhibit CENPF farnesylation in zygotes. The results showed that CENPF degradation was inhibited and its specific localization on nuclear membranes in morula and blastocyst vanished after FTI treatment. Also, CAAX motif mutation leads to failure of CENPF-C630 localization in morula and blastocyst. These results indicate that farnesylation plays a key role during CENPF degradation and localization in early embryos. To further assess CENPF function in parthenogenetic or fertilized embryos development, morpholino (MO) and Trim-Away were used to disturb CENPF function. CENPF knockdown in Metaphase II (MII) oocytes, zygotes or embryos with MO approach resulted in failure to develop into morulae and blastocysts, revealing its indispensable role in both parthenogenetic and fertilized embryos. Disturbing of CENPF with Trim-Away approach in zygotes resulted in impaired development of 2-cell and 4-cell, but did not affect the morula and blastocyst formation because of the recovered expression of CENPF. Taken together, our data suggest CENPF plays an important role during early embryonic development in mice. Abbreviation: CENPF: centromere protein F; MO: morpholino; FTI: Farnesyltransferase inhibitor; CENPE: centromere protein E; IVF: in vitro fertilization; MII: metaphase II; SAC: spindle assembly checkpoint; Mad1: mitotic arrest deficient 1; BUB1: budding uninhibited by benzimidazole 1; BUBR1: BUB1 mitotic checkpoint serine/threonine kinase B; Cdc20: cell division cycle 20.

Sodium nitrite negatively affects reproductive ability and offspring survival in female mice.

Sodium nitrite (NaNO2) is an industrial chemical that is frequently used as a food additive to prevent botulism and enhance glossiness, such as curing meat. In addition, in some regions, water source NaNO2 concentrations exceed standard regulatory levels. Whether the excessive intake of NaNO2 has toxic effects on female fertility and fetal development remain unknown. In this study, we administered ICR mice control saline, low-dose NaNO2 (60 mg/kg/day), or high-dose NaNO2 (120 mg/kg/day) by intragastric gavage for 21 days. We then assessed oocyte morphology, spindle-chromosome dynamics, mitochondrial distribution, ATP content, apoptotic cell numbers, DNA damage levels, histone modifications, reactive oxygen species (ROS) levels, and offspring survival. Results showed that NaNO2 treatment decreased oocyte number, impaired polar body extrusion, and increased zona pellucida thickness in oocytes. Furthermore, NaNO2 disrupted MII spindle integrity, caused abnormal mitochondrial distribution, decreased ATP content, and increased levels of ROS and H3K4me2. Moreover, the number of oocytes in early stages of apoptosis and with levels of DNA damage increased in NaNO2-treated mice along with decreased offspring numbers and survival rates. We demonstrated the negative effects of NaNO2 on female reproductive abilities in mice.

Glial cell line-derived neurotrophic factor supplementation promotes bovine in vitro oocyte maturation and early embryo development.

Paracrine factors such as glial cell line-derived neurotrophic factor (GDNF), which was originally derived from the supernatants of a rat glioma cell line, play pivotal roles in oocyte maturation and early embryo development in mammals, such as mice, rats, pigs, sheep, and even humans. However, whether GDNF facilitates in vitro oocyte maturation or early embryo development in bovines is not yet known. We show for the first time that GDNF and its receptor, GDNF family receptor alpha-1 (GFRA1), are presented in ovarian follicles at different stages as well as during oocyte maturation and early embryo development. Immunostaining results revealed the subcellular localizations of GDNF and GFRA1 in oocytes throughout follicle development, first in germinal vesicles and during blastocyst embryo stages. The ability of exogenously applied GDNF to promote oocyte maturation and early embryo development was evaluated in culture, where we found that an optimal concentration of 50 ng/mL promotes the maturation of cumulus-oocyte complexes and the nuclei of denuded oocytes as well as the development of embryos after IVF. To further investigate the potential mechanism by which GDNF promotes oocyte maturation, bovine oocytes were treated with morpholinos targeting Gfra1. The suppression of GFRA1 presence blocked endogenous and exogenous GDNF functions, indicating that the effects of GDNF that are essential and beneficial for bovine oocyte maturation and early embryo development occur through this receptor. Furthermore, we show that supplementation with GDNF improves the efficiency of bovine IVF embryo production.