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Alternative splicing expands the transcriptome and proteome complexity and plays essential roles in tissue development and human diseases. However, how alternative splicing regulates spermatogenesis remains largely unknown. Here, using a germ cell-specific knockout mouse model, we demonstrated that the splicing factor Srsf10 is essential for spermatogenesis and male fertility. In the absence of SRSF10, spermatogonial stem cells can be formed, but the expansion of Promyelocytic Leukemia Zinc Finger (PLZF)-positive undifferentiated progenitors was impaired, followed by the failure of spermatogonia differentiation (marked by KIT expression) and meiosis initiation. This was further evidenced by the decreased expression of progenitor cell markers in bulk RNA-seq, and much less progenitor and differentiating spermatogonia in single-cell RNA-seq data. Notably, SRSF10 directly binds thousands of genes in isolated THY+ spermatogonia, and Srsf10 depletion disturbed the alternative splicing of genes that are preferentially associated with germ cell development, cell cycle, and chromosome segregation, including Nasp, Bclaf1, Rif1, Dazl, Kit, Ret, and Sycp1. These data suggest that SRSF10 is critical for the expansion of undifferentiated progenitors by regulating alternative splicing, expanding our understanding of the mechanism underlying spermatogenesis.

Precise regulation of chromosome separation through spindle assembly checkpoint (SAC) during oocyte meiosis is critical for mammalian reproduction. The kinetochore plays an important role in the regulation of SAC through sensing microtubule tension imbalance or missing microtubule connections. Here, we report that kinetochore scaffold 1 (KNL1, also known as CASC5), an outer kinetochore protein, plays a critical role in the SAC function of mouse oocytes. KNL1 localized at kinetochores from GVBD to the MII stage, and microinjection of KNL1-siRNA caused accelerated metaphase-anaphase transition and premature first meiosis completion, producing aneuploid eggs. The SAC was prematurely silenced in the presence of unstable kinetochore-microtubule attachments and misaligned chromosomes in KNL1-depleted oocytes. Additionally, KNL1 and MPS1 had a synergistic effect on the activation and maintenance of SAC. Taken together, our results suggest that KNL1, as a kinetochore platform protein, stabilizes SAC to ensure timely anaphase entry and accurate chromosome segregation during oocyte meiotic maturation.

Calciphylaxis is a rare disease characterized histologically by microvessel calcification and microthrombosis, with high mortality and no proven therapy. Here, we reported a severe uremic calciphylaxis patient with progressive skin ischemia, large areas of painful malodorous ulcers, and mummified legs. Because of the worsening symptoms and signs refractory to conventional therapies, treatment with human amnion-derived mesenchymal stem cells (hAMSCs) was approved. Preclinical release inspections of hAMSCs, efficacy, and safety assessment, including cytokine secretory ability, immunocompetence, tumorigenicity, and genetics analysis in vitro, were introduced. We further performed acute and long-term hAMSC toxicity evaluations in C57BL/6 mice and rats, abnormal immune response tests in C57BL/6 mice, and tumorigenicity tests in neonatal Balbc-nu nude mice. After the preclinical research, the patient was treated with hAMSCs by intravenous and local intramuscular injection and external supernatant application to the ulcers. When followed up to 15 months, the blood-based markers of bone and mineral metabolism improved, with skin soft tissue regeneration and a more favorable profile of peripheral blood mononuclear cells. Skin biopsy after 1-month treatment showed vascular regeneration with mature noncalcified vessels within the dermis, and 20 months later, the re-epithelialization restored the integrity of the damaged site. No infusion or local treatment-related adverse events occurred. Thus, this novel long-term intravenous combined with local treatment with hAMSCs warrants further investigation as a potential regenerative treatment for uremic calciphylaxis due to effects of inhibiting vascular calcification, stimulating angiogenesis and myogenesis, anti-inflammatory and immune modulation, multidifferentiation, re-epithelialization, and restoration of integrity.

Serum uric acid level has been found to be associated with cerebrovascular diseases. However, whether serum uric acid level is a risk factor for arterial stiffness in the hypertension population is unclear. This study was designed to determine the relationship between serum uric acid level and arterial stiffness in the hypertension population. A total of 10450 participants were evaluated for the risk of arterial stiffness. Brachial-ankle pulse wave velocity (baPWV) was assessed, and high baPWV was determined as the highest quartile of baPWV values in a sex-specific manner. We evaluated the association between serum uric acid level and baPWV through multivariate-adjusted linear and logistic regression analyses. There was a significant difference on high baPWV between patients with quartiles of serum uric acid level in females and males (p

Oocyte meiotic maturation failure and unfaithful chromosome segregation are major causes for female infertility. Here, we showed that CENP-W, a relatively novel member of the kinetochore protein family, was expressed in mouse oocytes from the germinal vesicle (GV) to metaphase II (MII) stages. Confocal microscopy revealed that CENP-W was localized in the germinal vesicle in the GV stage, and then became concentrated on kinetochores during oocyte maturation. Knockdown of CENP-W by specific siRNA injection in vitro caused kinetochore-microtubule detachment, resulting in severely defective spindles and misaligned chromosomes, leading to metaphase I arrest and failure of first polar body (PB1) extrusion. Correspondingly, spindle assembly checkpoint (SAC) activation was observed in CENP-W knockdown oocytes even after 10h of culture. Our results suggest that CENP-W acts as a kinetochore protein, which takes part in kinetochore-microtubule attachment, thus mediating the progression of oocyte meiotic maturation.

Mammalian cyclin A1 is prominently expressed in testis and essential for meiosis in the male mouse, however, it shows weak expression in ovary, especially during oocyte maturation. To understand why cyclin A1 behaves in this way in the oocyte, we investigated the effect of cyclin A1 overexpression on mouse oocyte meiotic maturation. Our results revealed that cyclin A1 overexpression triggered meiotic resumption even in the presence of germinal vesicle breakdown inhibitor, milrinone. Nevertheless, the cyclin A1-overexpressed oocytes failed to extrude the first polar body but were completely arrested at metaphase I. Consequently, cyclin A1 overexpression destroyed the spindle morphology and chromosome alignment by inducing premature separation of chromosomes and sister chromatids. Therefore, cyclin A1 overexpression will prevent oocyte maturation although it can promote meiotic resumption. All these results show that decreased expression of cyclin A1 in oocytes may have an evolutional significance to keep long-lasting prophase arrest and orderly chromosome separation during oocyte meiotic maturation.

OTSSP167 is an anti-tumor drug significantly inhibiting tumor growth in xenotransplantation studies using mouse breast, lung, prostate, and pancreatic cancer cell lines. Its phase I clinical trial has been completed, indicating its great potential for future treatment of solid tumors. However, its drug-related adverse effects on reproductive systems have not yet been reported. In this study, we evaluated the effects of OTSSP167 on reproduction of female mice by determining oocyte quality and follicular development. We selected four-week-old female ICR mice for a 21-day intraperitoneal injection of OTSSP167 at a dose of 5 mg/kg/d. We found that OTSSP167 could block the meiotic process of oocytes, leading to a decrease in oocyte maturation and ovulated oocyte numbers, as well as a decrease in the quality of oocytes. The results showed that OTSSP167 treatment caused disordered spindle assembly, decreased mitochondria membrane potential, and increased accumulation of reactive oxygen species in oocytes. Further investigation showed that OTSSP167 induced DNA double-strand breaks, as indicated by increased levels of γH2AX in oocytes of primordial follicles and granulosa cells of growing follicles, which induced follicular atresia and decreased the numbers of follicles at various growing stages. Our study suggests that OTSSP167 treatment may have serious effects on the ovary and consequences for female cancer patients, providing strong evidence for the necessity of protecting female fertility in clinical OTSSP167 trials.

Protein phosphatase 6 (PP6) is a member of the PP2A-like subfamily, which plays a critical role in many fundamental cellular processes. We recently reported that PP6 is essential for female fertility. Here, we report that PP6 is involved in meiotic recombination and that germ cell-specific deletion of PP6 by Stra8-Cre causes defective spermatogenesis. The PP6-deficient spermatocytes were arrested at the pachytene stage and defects in DSB repair and crossover formation were observed, indicating that PP6 facilitated meiotic double-stranded breaks (DSB) repair. Further investigations revealed that depletion of PP6 in the germ cells affected chromatin relaxation, which was dependent on MAPK pathway activity, consequently preventing programmed DSB repair factors from being recruited to proper positions on the chromatin. Taken together, our results demonstrate that PP6 has an important role in meiotic recombination and male fertility.

Calciphylaxis is a rare disease characterized histologically by microvessel calcification and microthrombosis, with high mortality and no proven therapy. We reported a severe uremic calciphylaxis patient with progressive skin ischemia, large areas of painful malodorous ulcers and mummified legs. Because of her rapid progression and refractory to conventional therapy, human amnion-derived mesenchymal stem cells (hAMSCs) treatment was approved. Establishment and release inspection of hAMSCs, efficacy and safety assessment including cytokines secretory ability, immunocompetence, tumorigenicity and genetics analysisin vitrowere introduced. We further performed acute and long-term hAMSC toxity evaluations in C57BL/6 mice/rats, abnormal immune response tests in C57BL/6 mice and tumorigenic tests in the neonatal NU nude mice. After pre-clinical research, she was treated by hAMSCs with intravenous and local intramuscular injection and external supernatants application to her ulcers. When followed up to 15 months, her blood-based markers of bone and mineral metabolism were improved, with regeneration of skin soft tissue and a more favorable profile of peripheral blood mononuclear cells. Skin biopsy after 1 month treatment showed vascular regeneration with mature non-calcified vessels within dermis and 20 months later re-epithelialization restored the integrity of damaged site. No infusion or local treatment related adverse events occurred. To the best of our knowledge, this is the first evidence for the clinical use of hAMSCs. These findings suggest hAMSCs warrant further investigation as a potential regenerative treatment for uremic calciphylaxis with effects of inhibiting vascular calcification, stimulating angiogenesis and myogenesis, anti-inflammatory and immune modulation, multi-differentiation, re-epithelialization and restorage of integrity.

A multilayered carbon–-Polytetrafluoroethylene/graphite sheet (C-PTFE/GS) coating is synthesized on a Ti plate by hydrothermal and immersion method. The innermost layer is composed of amorphous carbon and the outermost layer is made of a compound with PTFE, graphite sheets and nanotube. Interfacial contact resistance (ICR) decreases to 8.9 mΩ·cm−2 at the applied force of 1.4 MPa between carbon paper and the substrate with coating. The corrosive current density is only 0.49 μA/cm2 at the cathode, while the potential of 0.6 V is applied. The charge transfer resistance of the multilayer carbon coating is higher than that of a bare sample through the results of electrochemical impedance spectroscopy. Moreover, there are no obvious changes in the coating before and after potentiostatic polarization testing by X-ray photoelectron spectroscopy (XPS) analysis, which demonstrates the stability of multilayered C-PTFE/GS coating on Ti. In addition, the surface morphology of C-PTFE/GS coating is preserved without defect after potentiostatic polarization in a simulated environment of proton exchange membrane fuel cells (PEMFCs). Therefore, the C-PTFE/GS coating is potentially applied as bipolar plates in PEMFCs.

Previous studies reported that microRNA-1298 was abnormally expressed in the myocardium of rat hearts after hypoxia/normoxia injury. This study aims to investigate the function and specific mechanism of miR-1298 in myocardial ischemia/reperfusion (IR) injury. Neonatal rat cardiomyocytes (NRCMs) were isolated from neonatal rat hearts and subjected to oxygen/glucose deprivation/reperfusion (OGD/R) to induce I/R injury. The rat model with I/R injury was induced by ligating the proximal left anterior descending artery (LAD). MiR-1298 expression was detected by qRT-PCR. The levels of PP2A, Bcl-2, Bax, and AMPK signaling members (p-AMPK, p-GSK3β) was detected by Western blot. Cell apoptosis was evaluated by TUNEL staining assay and flow cytometry. The infarct size of rat hearts was assessed by TTC staining assay. Premature and mature MiR-1298 were significantly downregulated while PP2A was significantly upregulated during I/R injury both in vitro and in vivo. The prediction of Starbase suggested that PP2A was a potential target of miR-1298. MiR-1298 overexpression significantly reduced cardiomyocyte apoptosis in vitro, and its protective effect was obviously attenuated by PP2A overexpression. Luciferase reporter assay showed that miR-1298 targeted PP2A directly. In addition, miR-1298 overexpression significantly reduced infarct size and cardiomyocyte apoptosis in the hearts of rats received with I/R injury in vivo. Moreover, miR-1298 overexpression significantly elevated the levels of Bcl-2 and AMPK signaling members (p-AMPK, p-GSK3β) while decreased Bax level, and these effects were partially reversed by PP2A overexpression. MiR-1298 participated in myocardial I/R injury by targeting the PP2A/AMPK/GSK3β signaling pathway, suggesting that miR-1298 might be a potential therapeutic target for myocardial I/R injury.

The quality of oocytes is a vital factor for embryo development. Meiotic progression through metaphase I usually takes a relatively long time to ensure correct chromosome separation, a process that is critical for determining oocyte quality. Here, we report that cell division cycle 5-like (Cdc5L) plays a critical role in regulating metaphase-to-anaphase I transition during mouse oocyte meiotic maturation. Knockdown of Cdc5L by small interfering RNA injection did not affect spindle assembly but caused metaphase I arrest and subsequent reduced first polar body extrusion due to insufficient anaphase-promoting complex/cyclosome activity. We further showed that Cdc5L could also directly interact with securin, and Cdc5L knockdown led to a continuous high expression level of securin, causing severely compromised meiotic progression. The metaphase-to-anaphase I arrest caused by Cdc5L knockdown could be rescued by knockdown of endogenous securin. In summary, we reveal a novel role for Cdc5L in regulating mouse oocyte meiotic progression by interacting with securin.

In mammals, oocytes are arrested at G2/prophase for a long time, which is called germinal vesicle (GV) arrest. After puberty, fully-grown oocytes are stimulated by a gonadotropin surge to resume meiosis as indicated by GV breakdown (GVBD). CCNB1 is accumulated to a threshold level to trigger the activation of maturation promoting factor (MPF), inducing the G2/M transition. It is generally recognized that the anaphase-promoting complex/cyclosome (APC/C) and its cofactor CDH1 (also known as FZR1) regulates the accumulation/degradation of CCNB1. Here, by using small interfering RNA (siRNA) and messenger RNA (mRNA) microinjection, immunofluorescence and confocal microscopy, immunoprecipitation, time-lapse live imaging, and immunoblotting analysis, we showed that Septin 4 regulates the G2/M transition by regulating the accumulation of CCNB1 via APC/C

In the microelectromechanical systems area, micrometal devices can be fabricated by microelectroforming process. However, since the defect mechanism of crystal growth, the electroformed layer suffers from the large internal compressive stress. To reduce the large internal compressive stress, the effects of different current densities on the internal compressive stress are investigated. From the view of dislocation density, the mechanism to reduce the compressive stress by adjusting the current densities has been researched originally. The electroforming experiments were processed by several current densities. The dislocation density was measured by the X-ray diffraction method. The compressive stress was tested by side incline method. The nanocrystal structure was observed by the transmission electron microscopy method. The experimental results show that the dislocation density was decreased along with the decreasing of the current density, as well as the compressive stress. The mechanisms are that the crystal structures including the crystallite size, the microcrystal distortion and the dislocation density can be refined by adjusting the small current densities. Then, the compressive stress becomes lower. This work contributes to fabricate the electroformed layer with low internal compressive stress.

NEK5, a member of never in mitosis-gene A-related protein kinase, is involved in the regulation of centrosome integrity and centrosome cohesion at mitosis in somatic cells. In this study, we investigated the expression and function of NEK5 during mouse oocyte maturation and preimplantation embryonic development. The results showed that NEK5 was expressed from germinal vesicle (GV) to metaphase II (MII) stages during oocyte maturation with the highest level of expression at the GV stage. It was shown that NEK5 localized in the cytoplasm of oocytes at GV stage, concentrated around chromosomes at germinal vesicle breakdown (GVBD) stage, and localized to the entire spindle at prometaphase I, MI and MII stages. The small interfering RNA-mediated depletion of Nek5 significantly increased the phosphorylation level of cyclin-dependent kinase 1 in oocytes, resulting in a decrease of maturation-promoting factor activity, and severely impaired GVBD. The failure of meiotic resumption caused by Nek5 depletion could be rescued by the depletion of Wee1B. We found that Nek5 depletion did not affect CDC25B translocation into the GV. We also found that NEK5 was expressed from 1-cell to blastocyst stages with the highest expression at the blastocyst stage, and Nek5 depletion severely impaired preimplantation embryonic development. This study demonstrated for the first time that NEK5 plays important roles during meiotic G2/M transition and preimplantation embryonic development.

Family with sequence similarity 46, member C (FAM46C) is a highly conserved non-canonical RNA polyadenylation polymerase that is abundantly expressed in human and mouse testes and is frequently mutated in patients with multiple myeloma. However, its physiological role remains largely unknown. In this study, we found that FAM46C is specifically localized to the manchette of spermatids in mouse testes, a transient microtubule-based structure mainly involved in nuclear shaping and intra-flagellar protein traffic. Gene knockout of FAM46C in mice resulted in male sterility, characterized by the production of headless spermatozoa in testes. Sperm heads were intermittently found in the epididymides of FAM46C knockout mice, but their fertilization ability was severely compromised based on the results of intracytoplasmic sperm injection assays. Interestingly, our RNA-sequencing analyses of FAM46C knockout testes revealed that mRNA levels of only nine genes were significantly altered compared to wild-type ones (q < 0.05). When considering alternate activities for FAM46C, in vitro assays demonstrated that FAM46C does not exhibit protein kinase or AMPylation activity against general substrates. Together, our data show that FAM46C in spermatids is a novel component in fastening the sperm head and flagellum.

AIM To understand the agreement, precision, and accuracy between other estimated glomerular filtration rate (eGFR) equations and the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) creatinine-cystatin C equation (EPI_Cr_CysC). MATERIALS AND METHODS We conducted a cross-sectional study of 1,913 CKD patients. The eGFRs were calculated separately by creatinine clearance rate and Cockcroft-Gault equation corrected for standard body surface area (Ccr_BSA and eCcr_BSA); CKD-EPI creatinine equation (EPI_Cr); CKD-EPI cystatin C equation (EPI_CysC); EPI_Cr_CysC equation; Modification of Diet in Renal Disease (MDRD) Study equation with standardized serum creatinine; and full-age spectrum creatinine equation (FAS). The EPI_Cr_CysC equation was used as the reference. RESULTS When compared with the EPI_Cr_CysC equation, the EPI_Cr equation achieved the highest agreement in eGFRs (Lin's concordance correlation coefficient = 0.936, 95% confidence interval (CI) = 0.930, 0.941). eCcr_BSA and EPI_Cr equations achieved the first and second highest percentage agreement in the accurate classification of CKD stage (72.55 vs. 71.25%). The MDRD equation had minimal bias and was closely followed by the EPI_Cr equation (median difference = -1.3, 95% CI = -2.0, -0.8 vs. median difference = 2.5, 95% CI = 1.7, 3.3 mL/min/1.73m2). The EPI_CysC and EPI_Cr equations achieved the first and second highest precision (interquartile range (IQR) of the difference = 12.2, 95% CI = 11.6, 12.9 vs. IQR of the difference = 15.5, 95% CI = 14.7, 16.3 mL/min/1.73m2). The EPI_Cr and MDRD equations performed similarly and both had the highest accuracy at 30% (1 - P30 = 18.6, 95% CI = 16.9, 20.4 vs. 1 - P30 = 18.6, 95% CI = 16.8, 20.3%). CONCLUSION For assessment of renal function, the EPI_Cr equation performed the best and remained an acceptable alternative to the EPI_Cr_CysC equation in the absence of cystatin C. .

Ti and its alloys, because of its poor wear and oxidation resistance, have seriously restricted its industrial applications in the dry friction conditions. Herein we reported an easy-handling and effective approach for electroless Ni-LaCl3 nanocomposite films on nanoporous ATO surface of Ti substrate to modify its surface properties. Anodized titanium oxide, shorted as ATO, was designed as a template to deposit Ni-LaCl3 films on Ti surface with superior bonding interface. By adjusting the anodizing voltage from DC 60 to 240 V, the surface of ATO templates was manipulated to achieve a suitable diameter of nanopores using high voltage with low-current density state. The as-received Ni-LaCl3 film with a leaflike surface was successfully formed on to ATO surface. With an increase in LaCl3 addition from 0 to 4.0 g L−1 in bath, results indicated that the diversified orientations of Ni crystals along Ni (111), (200), (220) and (311) were detected for Ni-LaCl3 films, rather than preferred directions of Ni (111) and (200) for pure Ni sample. As expected, the specific wear rate was ~ 10−5 mm3 (N m)−1 for Ni-4.0 g L−1 LaCl3 films, which was two times lower than that of the un-coated Ti substrate. Besides a lower microhardness of ~ 629 HV0.2 was detected for pure Ni, which was much lower that of ~ 752 HV0.2 for Ni-LaCl3 films, but higher than that of ~ 336 HV0.2 for Ti substrate. In addition, a superior corrosion resistance was obtained for Ni-LaCl3 composites relative to pure Ni, which was ascribed to the coexistence of La3+ ions and its La-rich insoluble products on its corroded surfaces for completing the pitting holes. In view of these, the as-deposited Ni-LaCl3 films on the surface of Ti substrate could guide an available proposal for surface modifying of Ti alloys against the wear corrosive failures, further intensifying their servicing life as subjected to harsh conditions.

The quality of post-ovulatory oocytes decreases with aging. In this study, we aimed to investigate the effects of N-acetyl-L-cysteine (NAC), a broadly used antioxidant, on oocyte quality in mouse post-ovulatory oocyte aging in vitro. NAC at 0.6mM concentration was added to culture medium (M2), and the quality of oocytes was analyzed at 6h, 12h, 18h and 24h of culture. We found that the frequency of spindle defects decreased in NAC-treated oocytes compared to those without NAC treatment. NAC treatment significantly decreased abnormal distribution of cortical granules (CGs) in oocytes during aging for 18h and 24h. Decreased intracellular reactive oxygen species (ROS) was also observed. Increased intracellular ATP levels and decreased abnormal distribution of mitochondria could be observed with NAC supplementation during post-ovulatory oocyte aging in vitro. These results indicate that NAC will maintain the quality of oocytes, and delay post-ovulatory oocyte aging as studied in the mouse.

PKCβI, a member of the classical protein kinase C family, plays key roles in regulating cell cycle transition. Here, we report the expression, localization and functions of PKCβI in mouse oocyte meiotic maturation. PKCβI and p-PKCβI (phosphor-PKCβI) were expressed from germinal vesicle (GV) stage to metaphase II (MII) stage. Confocal microscopy revealed that PKCβI was localized in the GV and evenly distributed in the cytoplasm after GV breakdown (GVBD), and it was concentrated at the midbody at telophase in meiotic oocytes. While, p-PKCβI was concentrated at the spindle poles at the metaphase stages and associated with midbody at telophase. Depletion of PKCβI by specific siRNA injection resulted in defective spindles, accompanied with spindle assembly checkpoint activation, metaphase I arrest and failure of first polar body (PB1) extrusion. Live cell imaging analysis also revealed that knockdown of PKCβI resulted in abnormal spindles, misaligned chromosomes, and meiotic arrest of oocytes arrest at the Pro-MI/MI stage. PKCβI depletion did not affect the G2/M transition, but its overexpression delayed the G2/M transition through regulating Cyclin B1 level and Cdc2 activity. Our findings reveal that PKCβI is a critical regulator of meiotic cell cycle progression in oocytes. Abbreviations: PKC, protein kinase C; COC, cumulus-oocyte complexes; GV, germinal vesicle; GVBD, germinal vesicle breakdown; Pro-MI, first pro-metaphase; MI, first metaphase; Tel I, telophase I; MII, second metaphase; PB1, first polar body; SAC, spindle assembly checkpoint.

Purpose: Accurate preoperative parathyroid localization is important for successful parathyroidectomy (PTX). The aim of our study was to investigate whether SPECT/CT has enhanced effect in preoperative localization of parathyroids. Methods: In our retrospective cohort study, we evaluated the effects of technetium-99m methoxyisobutylisonitrile-single-photon emission computed tomography/computed tomography (99mTc-MIBI SPECT/CT) on preoperative parathyroid localization for 645 secondary hyperparathyroidism (SHPT) patients. Among them, 569 successful PTX patients were divided into group A (received 99mTc-MIBI scintigraphy, n = 175) and group B (received 99mTc-MIBI scintigraphy and SPECT/CT imaging, n = 394). Sensitivity, specificity, and consistency of two imaging methods in preoperative localization of parathyroids were compared. Results: Overall sensitivity and consistency were higher in group B, while there was no difference in specificity between the two groups. In group A, the sensitivity of 99mTc-MIBI was 50.00%, 77.11%, 61.76%, and 76.54% in the right upper gland (RU), right lower gland (RL), left upper gland (LU), and left lower gland (LL) subgroups, while the consistency was 52.00%, 76.57%, 61.71%, and 75.43%, respectively. In group B, the sensitivity of 99mTc-MIBI with SPECT/CT was 69.39%, 90.03%, 78.07%, and 84.27%, and the consistency was 69.54%, 88.32%, 78.43%, and 84.26%, respectively. The sensitivity and consistency in lower glands were higher than in upper glands in both groups. Sensitivity for eutopic parathyroid was higher in group B, while there was no difference for ectopic parathyroid. Conclusions: 99mTc-MIBI SPECT/CT can increase the sensitivity and consistency of preoperative localization of eutopic parathyroid glands, and it can accurately locate ectopic parathyroid without sensitivity improvement.

Microtubule plus-end tracking proteins (+TIPs) associate with growing microtubule plus ends and control microtubule dynamics and interactions with different cellular structures during cell division, cell migration and morphogenesis. Microtubule-associated RP/EB family member 2 (MAPRE2/EB2) is a highly conserved core component of +TIPs networks, but whether this molecule is required for mammalian meiotic progression is unknown. In this study, we investigated the expression and function of MAPRE2 during oocyte maturation. Our results showed that MAPRE2 was consistently expressed from germinal vesicle (GV) to metaphase II (MII) stages and that MAPRE2 was distributed in the cytoplasm of oocytes at GV stage and along the spindle at metaphase I (MI) and MII stages. Small interfering RNA-mediated knockdown of Mapre2 severely impaired microtubule stability, kinetochore-microtubule attachment, and chromosome alignment and subsequently caused spindle assembly checkpoint (SAC) activation and cyclin B1 nondegradation, leading to failure of chromosome segregation and first polar body extrusion. This study demonstrates for the first time that MAPRE2 plays an important role during mouse oocyte meiosis.

A highly corrosion resistant and conductive carbon/PTFE/TiN composite coating on Ti bipolar plates is prepared by a two-step hydrothermal and impregnation process. The composite coating exhibits a full coverage with uniformly distributed PTFE and TiN particles when 0.1 mol/L of glucose is used as carbon source and 10 wt % PFTE + 3 g/L TiN suspension as impregnating solution. The as-prepared composite coating film shows 0.009 μA cm−2 of corrosion current density and 13 mΩ cm2 of ICR in the simulated solution of proton exchange membrane fuel cells, respectively. Meanwhile, the potentiostatic polarization test shows that the corrosion current density of the composite coating is below 1 μA cm−2 under real PEMFC working conditions of the cathode (0.6 V) and anode (−0.1 V). Additionally, the composite coating enhances hydrophobic property with a static contact angle of 120.2°and 115.5°before and after polarization test, respectively. In summary, the Ti substrate with carbon/PTFE/TiN composite coating shows a great potential application as bipolar plates of PEMFCs.

There are two important events in oocyte meiotic maturation, the G2/M transition and metaphase I progression. Thousands of proteins participate in regulating oocyte maturation, which highlights the importance of the ubiquitin proteasome system (UPS) in regulating protein synthesis and degradation. Skp1–Cullin–F-box (SCF) complexes, as the best characterized ubiquitin E3 ligases in the UPS, specifically recognize their substrates. F-box proteins, as the variable adaptors of SCF, can bind substrates specifically. Little is known about the functions of the F-box proteins in oocyte maturation. In this study, we found that depletion of FBXO34, an F-box protein, led to failure of oocyte meiotic resumption due to a low activity of MPF, and this phenotype could be rescued by exogenous overexpression of CCNB1. Strikingly, overexpression of FBXO34 promoted germinal vesicle breakdown (GVBD), but caused continuous activation of spindle assembly checkpoint (SAC) and MI arrest of oocytes. Here, we demonstrated that FBXO34 regulated both the G2/M transition and anaphase entry in meiotic oocytes.

Workforce analytics brings data-driven methods to organizations for deriving insights from employee-related data and supports decision making. However, it faces an open challenge of lacking the capability to analyze the behavior of employee groups in order to understand organizational performance. This paper proposes a novel notion of work profiles of resource groups, informed by the management literature, for characterizing resource group behavior from multiple aspects relevant to workforce performance. This notion is central to the design of a new, systematic approach that supports resource group analysis by exploiting business process execution data. The approach also provides managers and business analysts with an intuitive means of group-oriented resource analysis by applying visual analytics. We demonstrate the applicability of the approach and usefulness of the proposed notion of resource group work profiles using real datasets from five Dutch municipalities.

Predictive process analytics focuses on predicting the future states of running instances of a business process. While advanced machine learning techniques have been used to increase the accuracy of predictions, the resulting predictive models lack transparency. Explainable machine learning methods can be used to interpret black-box models. However, it is unclear how fit for purpose these methods are in explaining process predictive models. In this paper, we aim to investigate the capabilities of two explainable methods, LIME and SHAP, in reproducing the decision-making processes of black-box process predictive models. We focus on fidelity metrics and propose a method to evaluate the faithfulness of LIME and SHAP when explaining process predictive models built on a Gradient Boosting Machine classifier. We conduct the evaluation using three real-life event logs and analyze the fidelity evaluation results to derive insights. The research contributes to evaluating the trustworthiness of explainable methods for predictive process analytics as a fundamental and key step towards human user-oriented evaluation.

Background: Multiple studies showed that long-chain noncoding RNA H19 (LncRNA H19) is high-expressed in human and mouse abdominal aortic aneurysms (AAAs). We speculated that it plays an important role in arterial disease, and therefore studied the role and mechanism of H19 in aortic dissection (AD). Methods: The expressions of related genes in human aortic smooth muscle cells (HASMCs) induced by platelet-derived growth factor BB (PDGF-BB) or in the aortic tissue of AD patients/mice were identified by Western blot and quantitative real-time polymerase chain reaction. The targeting relationship between H19 and miR-193b-3p was predicted and verified by bioinformatics analysis, dual luciferase assay, RNA pull-down assay, RNA immunoprecipitation (RIP), and Pearson correlation coefficient. The H19 and miR-193b-3p effects on the biological functions of tissues and cells were examined by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, thiazolyl blue tetrazolium bromide) assay, wound-healing assay, and Hematoxylin–Eosin (HE) staining. Results: LncRNA H19 was abnormally high-expressed in thoracic aorta tissues of AD patients, and it could competitively bind to and inhibit miR-193b-3p. In the PDGF-BB group, the expressions of H19, matrix metallopeptidase (MMP) 2 (MMP-2) and MMP-9 were up-regulated and the expressions of miR-193b-3p, α-SMA, and SM22α were down-regulated; moreover, the proliferation and migration rate of HASMCs were increased. However, H19 silencing reversed the regulation of PDGF-BB on HASMCs. More interestingly, miR-193b-3p inhibitor could partially reverse the effect of H19 silencing. In addition, the above results were verified by animal experiments, showing that shH19 and up-regulated miR-193b-3p could significantly reduce the thoracic aorta pathological damage in AD mice. Conclusion: LncRNA H19 regulated smooth muscle cell function by sponging miR-193b-3p and it participated in the development of AD.

Predictive process analytics uses advanced machine learning techniques to accurately predict future states of running business processes. Given the complexity of these predictive models, explainable AI techniques are also required to enable informed decision-making. However, few studies evaluate the quality of explanations provided by existing methods to explain business process predictions. In this paper, we attempt to evaluate the consistency of explanations produced for process predictions by two popular explainable methods. We propose that methods and metrics to assess feature selection algorithms can be used to evaluate explanation stability. We use these metrics to assess explanations produced by LIME and SHAP. Our findings indicate that explanation stability may depend on dataset characteristics, feature construction methods and predictive model characteristics. In addition, we also find that, though stable explanations are needed for informed decision-making, unexpected behaviour in explanation stability can act as a diagnostic tool to determine model quality.

Gefitinib is a first-line anti-cancer drug for the treatment of advanced non-small cell lung cancer (NSCLC). It has been reported that gefitinib can generate several drug-related adverse effects, including nausea, peripheral edema, decreased appetite and rash. However, the reproductive toxicity of gefitinib has not been clearly defined until now. Here we assessed the effects of gefitinib on oocyte quality by examining the critical events and molecular changes of oocyte maturation. Gefitinib at 1, 2, 5 or 10 μM concentration was added to culture medium (M2). We found that gefitinib at its median peak concentration of 1 μM did not affect oocyte maturation, but 5 μM gefitinib severely blocked oocyte meiotic progression as indicated by decreased rates of germinal vesicle breakdown (GVBD) and polar body extrusion (PBE). We further showed that gefitinib treatment increased phosphorylation of CDK1 at the site of Try15, inhibited cyclin B1 entry into the nucleus, and disrupted normal spindle assembly, chromosome alignment and mitochondria dynamics, finally leading to the generation of aneuploidy and early apoptosis of oocytes. Our study reported here provides valuable evidence for reproductive toxicity of gefitinib administration employed for the treatment of cancer patients.

To compare the clinical effect of low molecular weight heparin (LMWH) and regional citrate anticoagulation (RCA) in continuous renal replacement therapy (CRRT) after heart valve replacement.Retrospective analysis of clinical data from 60 patients undergoing continuous renal replacement therapy after heart valve replacement in Peking University Shenzhen Hospital from January 2015 to July 2019, including 35 males and 25 females, aged (58.83 ±16.36) years. The patients were divided into a LMWH group and a RCA group according to the anticoagulation regimen, with 30 patients in each group. The 24 h postoperative drainage volume (pericardium, mediastinum), postoperative ventilator use time and ICU stay time, incidence of postoperative respiratory tract bleeding and gastrointestinal bleeding, postoperative skin ecchymosis incidence and mortality between the 2 groups were compared; the thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), prothrombin time (PT), international normalized ratio (INR), and PLT between the 2 groups after anticoagulation treatment were compared; the service life of dialysis filters and CRRT time between the 2 groups were compared; and the levels of urea nitrogen and serum creatinine between the 2 groups before and after treatment were also compared.After anticoagulant treatment, the RCA group had less 24 h postoperative drainage volume than the LMWH group, shorter ventilator use time, ICU stay time, and hospitalization days than the LMWH group, and lower respiratory tract bleeding and gastrointestinal bleeding incidence than the LMWH group, with significant differences (allCRRT after cardiac valve replacement with RCA can effectively reduce pericardial and mediastinal drainage, reduce the risk of bleeding, shorten the time of ventilator use and ICU stay, and promote postoperative recovery of patients, which have positive significance for reducing the burden on patients and their families.

RNA-binding proteins (RBPs) have essential functions during germline and early embryo development. However, current methods are unable to identify the in vivo targets of a RBP in these low-abundance cells. Here, by coupling RBP-mediated reverse transcription termination with linear amplification of complementary DNA ends and sequencing, we present the LACE-seq method for identifying RBP-regulated RNA networks at or near the single-oocyte level. We determined the binding sites and regulatory mechanisms for several RBPs, including Argonaute 2 (Ago2), Mili, Ddx4 and Ptbp1, in mature mouse oocytes. Unexpectedly, transcriptomics and proteomics analysis of Ago2-/- oocytes revealed that Ago2 interacts with endogenous small interfering RNAs (endo-siRNAs) to repress mRNA translation globally. Furthermore, the Ago2 and endo-siRNA complexes fine-tune the transcriptome by slicing long terminal repeat retrotransposon-derived chimeric transcripts. The precise mapping of RBP-binding sites in low-input cells opens the door to studying the roles of RBPs in embryonic development and reproductive diseases.

Cadmium (Cd) is a bioaccumulative heavy metal element with potential toxicity on the female reproductive system, but the exact molecular mechanisms have not yet been clearly defined. In this study, female mice were exposed to 0.5 mg/kg/day of CdCl2 for 60 consecutive days. We found that chronic Cd exposure significantly decreased the fecundity of female mice by affecting oocyte meiotic progression as indicated by disrupted spindle assembly, chromosome alignment and kinetochore-microtubule attachments, consequently resulting in aneuploid oocytes. Further studies showed that the periodic fluctuations of MPF activity and cyclin B1 expression were disturbed in Cd-exposed oocytes probably by affecting the spindle assembly checkpoint protein Bub3. In addition, Cd exposure induced oxidative stress as indicated by an increased level of reactive oxygen species and apoptosis in oocytes, leading to oocyte quality deterioration. Taken together, these data suggest that Cd exposure causes disrupted molecular events of meiotic progression and deterioration of oocyte quality via oxidative stress, leading to decrease of female fertility.

Background: Non-dipping heart rate (HR), defined as night/day HR > 0.90, was associated with increased mortality in epidemiological studies. However, its prognostic value in stage 5 chronic kidney disease (CKD5) patients remains unknown. Methods: This case-control study of 162 healthy controls and 502 CKD5 patients, including 316 secondary hyperparathyroidism (SHPT) with parathyroidectomy (PTX) subgroup were enrolled between 2011 and 2018. Each participant underwent 24-hour Holter for HR ratio. Mortality was followed up in CKD5 patients (median time: 46.0 months). Findings: HR ratio in CKD5 patients was higher than controls (0.92±0.08 vs 0.81±0.08, P 500.0 pg/ml). Interpretation: CKD5 patients displayed non-dipping HR pattern which is a prognostic marker of all-cause mortality. PTX for SHPT patients was associated with reversal in non-dipping ratio which may mediate a better outcome. Funding Statement: National Natural Science Foundation of China (81270408, 81570666), International Society of Nephrology Clinical Research Program (18-01-0247), Program of Jiangsu Clinical Research Center (BL2014084), Jiangsu Province Key Medical Personnel Project (ZDRCA2016002). Declaration of Interests: All the authors declared no competing interests. Ethics Approval Statement: All patients and controls participating in the study gave written informed consent, and the study protocol was approved by the Research Ethics Committee of the First Affiliated Hospital of Nanjing Medical University, Nanjing, China (Ethics approval number: 2011-SR-072, 2019-SR-368).

There has been a growing interest in the literature on the application of deep learning models for predicting business process behaviour, such as the next event in a case, the time for completion of an event, and the remaining execution trace of a case. Although these models provide high levels of accuracy, their sophisticated internal representations provide little or no understanding about the reason for a particular prediction, resulting in them being used as black-boxes. Consequently, an interpretable model is necessary to enable transparency and empower users to evaluate when and how much they can rely on the models. This paper explores an interpretable and accurate attention-based Long Short Term Memory (LSTM) model for predicting business process behaviour. The interpretable model provides insights into the model inputs influencing a prediction, thus facilitating transparency. An experimental evaluation shows that the proposed model capable of supporting interpretability also provides accurate predictions when compared to existing LSTM models for predicting process behaviour. The evaluation further shows that attention mechanisms in LSTM provide a sound approach to generate meaningful interpretations across different tasks in predictive process analytics.

SummarySynaptotagmin 1 (Syt1) is an abundant and important presynaptic vesicle protein that binds Ca2+ for the regulation of synaptic vesicle exocytosis. Our previous study reported its localization and function on spindle assembly in mouse oocyte meiotic maturation. The present study was designed to investigate the function of Syt1 during mouse oocyte activation and subsequent cortical granule exocytosis (CGE) using confocal microscopy, morpholinol-based knockdown and time-lapse live cell imaging. By employing live cell imaging, we first studied the dynamic process of CGE and calculated the time interval between [Ca2+]i rise and CGE after oocyte activation. We further showed that Syt1 was co-localized to cortical granules (CGs) at the oocyte cortex. After oocyte activation with SrCl2, the Syt1 distribution pattern was altered significantly, similar to the changes seen for the CGs. Knockdown of Syt1 inhibited [Ca2+]i oscillations, disrupted the F-actin distribution pattern and delayed the time of cortical reaction. In summary, as a synaptic vesicle protein and calcium sensor for exocytosis, Syt1 acts as an essential regulator in mouse oocyte activation events including the generation of Ca2+ signals and CGE.