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Abstract Background Attention deficit hyperactivity disorder (ADHD) and tic disorder (TD) are two common neurodevelopmental disorders that frequently occur in childhood, and these two disorders often coexist. Cluster analysis provides a novel perspective on the heterogeneity of these commonly observed clinical disorders. Methods We recruited patients with comorbid ADHD and TD from Beijing Children's Hospital between May 2022 and August 2023, collecting data on their symptoms and functional impairments. The number of clusters was determined using the elbow method, and K-means clustering was conducted. Fisher discriminant analysis and silhouette score were used for validation. Additionally, we assessed premonitory urge, strengths, and difficulties among groups. We also collected samples with ADHD alone and performed cluster analyses. Results The number of clusters for the ADHD comorbid TD sample was determined to be two by the elbow method. The results of the cluster analysis showed that the ADHD comorbid TD sample could be divided into the severe TD group and the severe ADHD group. The severe TD group exhibits more pronounced tic symptoms, yet their age, ADHD symptoms, and functional impairment are all significantly lower than those of the severe ADHD group. Compared to samples with ADHD alone, the distribution of age and functional impairment among individuals does not change with the addition of TD symptoms, maintaining a parallel relationship with the severity of ADHD symptoms. Conclusion Patients with co-occurring ADHD and TD can be classified into two clusters based on age, symptoms, and functional impairment. In clinical interventions for these patients, while ADHD may require more attention, it is also crucial to identify the core symptoms of the patients. The heterogeneity in clinical symptom presentations highlights the need for individualized treatment approaches.

Abstract Aims Non-suicidal self-injury (NSSI) is a serious issue that is increasingly prevalent among children and adolescents, especially in rural areas. Developing a suitable predictive model for NSSI is crucial for early identification and intervention. Methods This study included 2090 Chinese rural children and adolescents. Participants’ sociodemographic information, symptoms of anxiety as well as depression, personality traits, family environment and NSSI behaviors were collected through a questionnaire survey. Gender, age, grade, and all survey results except sociodemographic information were used as relevant factors for prediction. Support vector machines, decision tree and random forest models were trained and validated by the train set and valid set, respectively. The metrics of each model were tested and compared to select the most suitable one. Furthermore, the mean decrease Gini index was calculated to measure the importance of relevant factors. Results The prevalence of NSSI was 38.3%. Out of the 6 models assessed, the random forest model demonstrated the highest suitability in predicting the prevalence of NSSI. It achieved sensitivity, specificity, AUC, accuracy, precision, and F1 scores of 0.65, 0.72, 0.76, 0.70, 0.57, and 0.61, respectively. Anxiety and depression were the top two contributing factors in the prediction model. Neuroticism and conflict were the factors that contributed the most to personality traits and family environment, respectively, in terms of prediction. In addition, demographic factors contributed little to the prediction in this study. Conclusion This study focused on Chinese children and adolescents in rural areas and demonstrated the potential of using machine learning approaches in predicting NSSI. Our research complements the application of machine learning methods to psychiatric and psychological problems.

Abstract Background While Tourette syndrome (TS) and attention-deficit/hyperactivity disorder (ADHD) often co-occur, the nature of the relationship between their symptoms is not well understood. Network analysis of psychopathology allow for detailed examinations of symptom interactions, providing an effective approach to explore the patterns of comorbidity between TS and ADHD symptoms. Methods This study included 3,958 participants (male/female = 3,004/954, age mean ± SD = 8.60 ± 2.25 years). We collected data on TS symptoms using the Motor Tic, Obsessions and Compulsions, Vocal Tic Evaluation Survey (MOVES), and ADHD symptoms using the Swanson, Nolan, and Pelham Rating Scale-IV (SNAP-IV). Network analysis was employed to construct a combined network of TS and ADHD symptoms at the symptom level. We utilized the expected influence (EI) and bridge EI metrics to explore the core and bridge symptoms within the network. Results The network structure demonstrated a moderate number of non-zero connections between TS and ADHD symptoms, constituting 23.06% of all potential connections. Core symptoms in the comorbidity network included “Often has difficulty sustaining attention in tasks or play activities,” “Certain bad words or thoughts keep going through my mind,” and “Words come out that I can’t stop or control.” Bridging symptoms identified were “Words come out that I can’t stop or control,” “I do certain things like jumping or clapping over and over,” “I can’t control all my movements,” and “Often talks excessively.” Conclusion The core and bridging symptoms identified in this study serve as potential therapeutic targets for the treatment of TS and ADHD comorbidity in clinical children and adolescents.

BackgroundTic disorder, a chronic neurodevelopmental disorder that typically onsets during childhood, is characterized by sudden, involuntary, rapid, and non-rhythmic motor and vocal tics. Individuals with tic disorders often experience physical health issues. The purpose of our retrospective analysis was to elucidate the common comorbid physical diseases and mental disorders and their characteristics of outpatient children with tic disorders in a large public children’s hospital in China over the past 5 years.MethodsParticipants were outpatients with tic disorders who visited Beijing Children’s Hospital, from January 1, 2018 to December 31, 2022. After the inclusion screening, a total of 523,462 patient visits were included in the analysis. Based on the International Classification of Diseases, 10th Revision (ICD-10) diagnostic system, we employed descriptive statistical analysis to examine the frequently co-occurring somatic diseases in patients with tic disorders, as well as the influence of variables such as age and seasonal variation on these comorbidities.ResultsThe top five diseases of total outpatient visits were as follows: Respiratory diseases, Mental and behavioral disorders, Diseases of the eye and adnexal, Digestive disorders, Diseases of the skin and subcutaneous tissue. Among the top five comorbid disease system, the most commonly third- level classification of diseases were upper respiratory tract infections, attention deficit and hyperactivity disorder, conjunctivitis, dyspepsia, dermatitis. Respiratory system diseases experienced a peak in April, while the other four types of diseases reached their peak in August. Additionally, each disease system showed the lowest number of patient visits in February. Additional to the mental and behavioral disorders, the other four disease systems would experience a peak in medical visits between the ages of 4 and 6.ConclusionOur study highlighted the most common physical diseases and mental disorders in tic disorders, namely the respiratory diseases, specifically upper respiratory tract infections, and mental and behavioral disorders, with ADHD being the most common co-occurring condition.

Abstract Background Zinc oxide nanoparticle (ZnO NP) is one of the metal nanomaterials with extensive use in many fields such as feed additive and textile, which is an emerging threat to human health due to widely distributed in the environment. Thus, there is an urgent need to understand the toxic effects associated with ZnO NPs. Although previous studies have found accumulation of ZnO NPs in testis, the molecular mechanism of ZnO NPs dominated a decline in male fertility have not been elucidated. Results We reported that ZnO NPs exposure caused testicular dysfunction and identified spermatocytes as the primary damaged site induced by ZnO NPs. ZnO NPs led to the dysfunction of spermatocytes, including impaired cell proliferation and mitochondrial damage. In addition, we found that ZnO NPs induced ferroptosis of spermatocytes through the increase of intracellular chelatable iron content and lipid peroxidation level. Moreover, the transcriptome analysis of testis indicated that ZnO NPs weakened the expression of miR-342-5p, which can target Erc1 to block the NF-κB pathway. Eventually, ferroptosis of spermatocytes was ameliorated by suppressing the expression of Erc1. Conclusions The present study reveals a novel mechanism in that miR-342-5p targeted Erc1 to activate NF-κB signaling pathway is required for ZnO NPs-induced ferroptosis, and provide potential targets for further research on the prevention and treatment of male reproductive disorders related to ZnO NPs. Graphical Abstract

Objectives This research aims to explore the correlation between ABO blood groups and neoplasms in the head and neck region, specifically investigating the susceptibility of different blood groups to tumours.Design Systematic review and meta-analysis.Data sources CNKI, WANFANG DATA, PubMed, Web of Science and Embase databases were systematically searched from inception to January 2024.Eligibility criteria All studies of ABO blood type and head and neck tumours will be included in this study.Data extraction and synthesis A systematic literature review was performed using digital platforms on CNKI, WANFANG DATA, PubMed, Web of Science and Embase databases, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines for meta-analysis. Two authors independently extracted the data and assessed the quality of included studies. The Newcastle-Ottawa Scale was used to assess the quality of the included studies. Comparisons were made between blood types A, B, AB and their combined group versus O, along with subgroup analyses. Systematic analysis was performed by using Review Manager V.5.4 and Stata V.18 statistical software.Results 30 articles were included, involving 737 506 subjects, among which 21 382 were patients with head and neck tumours. The overall analysis indicated a significant association between type AB blood (OR 0.762, 95% CI 0.637 to 0.910) and a reduced risk of head and neck tumours. In the Caucasoid race, type A blood is significantly linked to an elevated likelihood of head and neck tumours (OR 1.353, 95% CI 1.076 to 1.702), while in the Mongoloid race, type AB blood is significantly linked to a reduced likelihood of developing tumours in the head and neck area (OR= 0.732, 95% CI 0.588 to 0.910). No significant associations were found in the subgroup analysis by gender. Regarding different types of cancer, type A blood is significantly associated with an increased risk of salivary gland tumours (OR 1.338, 95% CI 1.075 to 1.665), and type AB blood is significantly linked to a lower likelihood of nasopharyngeal carcinoma (OR 0.590, 95% CI 0.429 to 0.812).Conclusion A correlation exists between ABO blood groups and tumours in the head and neck region. However, the link between blood type and head and neck tumours requires further confirmation through more prospective studies.PROSPERO registration number CRD42024510176.

The Frizzleds (FZDs) receptors on the cell surface belong to the class F of G protein-coupled receptors (GPCRs) which are the major receptors of WNT protein that mediates the classical WNT signaling pathway and other non-classical pathways. Besides, the FZDs also play a core role in tissue regeneration and tumor occurrence. With the structure and mechanism of FZDs activation becoming clearer, a series of FZDs modulators (inhibitors and agonists) have been developed, with the hope of bringing benefits to the treatment of cancer and degenerative diseases. Most of the FZDs inhibitors (small molecules, antibodies or designed protein inhibitors) block WNT signaling through binding to the cysteine-rich domain (CRD) of FZDs. Several small molecules impede FZDs activation by targeting to the third intracellular domain or the transmembrane domain of FZDs. However, three small molecules (FZM1.8, SAG1.3 and purmorphamine) activate the FZDs through direct interaction with the transmembrane domain. Another type of FZDs agonists are bivalent or tetravalent antibodies which activate the WNT signaling via inducing FZD-LRP5/6 heterodimerization. In this article, we reviewed the FZDs modulators reported in recent years, summarized the critical molecules’ discovery processes and the elucidated relevant structural and pharmacological mechanisms. We believe the summaried molecular mechanisms of the relevant modulators could provide important guidance and reference for the future development of FZD modulators.

Tourette Syndrome (TS) is a neurodevelopmental disorder characterized by multiple motor and vocal tics, often accompanied by comorbid disorders. Optional treatments for patients with TS include behavioral therapy, pharmacotherapy, and neurostimulation techniques. Deep brain stimulation (DBS) has been considered a therapeutic approach for refractory TS and its comorbid symptoms. However, systematic comparison is necessary to understand the therapeutic effect of DBS among patients with TS with various comorbid symptoms, demographic characteristics, or stimulation targets. Consequently, our research aimed to assess the clinical efficacy of DBS in alleviating the symptoms of TS and its comorbidities. A systematic literature search was conducted across five databases: PubMed, Web of Science, MEDLINE, Embase, and PsycINFO. The primary outcome was the mean change in the global score of the Yale Global Tic Severity Scale (YGTSS), which assesses the severity of tics. The secondary outcomes included mean improvement of comorbid symptoms, such as obsessive-compulsive behaviors (OCB), depression symptoms and anxiety symptoms. In total, 51 studies with 673 participants were included in this meta-analysis. Overall, the DBS led to a significant improvement in tic symptoms (p ​

BackgroundSensory symptoms linked to tic disorder (TD) are challenging to quantify via self- or parent-reported measures. The current study aimed to develop a novel observer-rated semi-structured interview, namely, the Sensory Phenomenon Assessment Scale (SPAS), to aid clinical evaluation on symptoms of TD among children.MethodsTo test its psychometric properties, tic, premonitory urge (PU), and obsessive–compulsive symptoms (OCS) were also assessed in 223 children via the Yale Global Tic Severity Scale (YGTSS), the Premonitory Urge for Tic Scale (PUTS), and the Children’s Yale–Brown Obsessive–Compulsive Scale (CY-BOCS). Factor analysis and internal consistency test were carried out using data from TD-diagnosed individuals.ResultsGood internal consistency and test–retest reliability were observed. Criterion validity was established by significant correlations between the PUTS, the YGTSS, the CY-BOCS, and scores of the SPAS. Factor analyses supported a single-factor model of the SPAS, in which the five items each showed a factor loading above 0.6.ConclusionThis study demonstrated that the SPAS is reliable and valid and, thus, can serve as a good and concise measure of clinical symptoms among children and adolescents with TD.

Cancer, with high morbidity and high mortality, is one of the major burdens threatening human health globally. Intervention procedures via percutaneous puncture have been widely used by physicians due to its minimally invasive surgical approach. However, traditional manual puncture intervention depends on personal experience and faces challenges in terms of precisely puncture, learning-curve, safety and efficacy. The development of puncture interventional surgery robotic (PISR) systems could alleviate the aforementioned problems to a certain extent. This paper attempts to review the current status and prospective of PISR systems for thoracic and abdominal application. In this review, the key technologies related to the robotics, including spatial registration, positioning navigation, puncture guidance feedback, respiratory motion compensation, and motion control, are discussed in detail.

The zinc oxide nanoparticles (ZnO NPs) is one of the most extensively utilized metal oxide nanoparticles in biomedicine, human food, cosmetics and livestock farming. However, growing evidence suggests that there is a potential risk for humans and animals because of the accumulation of ZnO NPs in cells, which leads to cell death through several different pathways. Nevertheless, the effects of ZnO NPs on porcine granulosa cells (PGCs) and how ZnO NPs regulate the follicular cells are unknown. In this study, we aimed to elucidate the role of ZnO NPs in the porcine ovary by using PGCs. Firstly, we identified the characterization of ZnO NPs used in this study and the results showed that the size of ZnO NPs was 29.0 nm. The results also demonstrated that ZnO NPs impaired cell viability and decreased steroid hormone secretion in PGCs. In addition, ZnO NPs induced reactive oxygen species (ROS) production, leading to oxidative stress of PGCs. Meanwhile, ZnO NPs also triggered autophagy in PGCs by increasing the ratio of LC3-II/LC3-I, along with the expression of SQSTM1 and ATG7. Finally, the results from N-acetylcysteine (NAC) addition suggested that ZnO NPs promoted autophagy through the enhancement of ROS production. In summary, this study demonstrates that ZnO NPs impair the viability and function of PGCs through autophagy, which is regulated by ROS production.

Abstract Depression is a common psychiatric disorder with high prevalence and mortality rates as well as high risk of serious harm in adolescents that have significant negative impact on families and society. The feeding inhibitor Nesfatin-1 contributes to the regulation of stress and emotion. The purpose of this project was to compare the differences in the levels of Nesfatin-1 between adolescents with depression and healthy adolescents, and verify the association between the levels of Nesfatin-1 and severity of depression in adolescents. Adolescents with depression (n = 61) and healthy adolescents (n = 30) were evaluated. The Hamilton Depression Rating Scale (HAMD-17) was used to classify the adolescents with depression. Thirty-one and thirty-two was assigned to the mild-to-moderate (HAMD-17 ≤ 24) depression group and severe group (HAMD-17 > 24). Plasma Levels of Nesfatin-1 were measured by human ELISA Kit and differences among groups evaluated. Data were analyzed using the statistical software SPSS 23. HAMD-17 score was significantly higher in adolescents with depression than that in the healthy adolescents (P 0.05). Plasma levels of Nesfatin-1 increased with severity of depression in adolescents and may be useful as a biomarker of depression severity. Further studies are needed in future projects.

Manipulating cells at a small scale is widely acknowledged as a complex and challenging task, especially when it comes to cell grasping and transportation. Various precise methods have been developed to remotely control the movement of microrobots. However, the manipulation of micro-objects necessitates the use of end-effectors. This paper presents a study on the control of movement and grasping operations of a magnetic microrobot, utilizing only 3 pairs of electromagnetic coils. A specially designed microgripper is employed on the microrobot for efficient cell grasping and transportation. To ensure precise grasping, a bending deformation model of the microgripper is formulated and subsequently validated. To achieve precise and reliable transportation of cells to specific positions, an approach that combines an extended Kalman filter with a model predictive control method is adopted to accomplish the trajectory tracking task. Through experiments, we observe that by applying the proposed control strategy, the mean absolute error of path tracking is found to be less than 0.155 mm. Remarkably, this value accounts for only 1.55% of the microrobot’s size, demonstrating the efficacy and accuracy of our control strategy. Furthermore, an experiment involving the grasping and transportation of a zebrafish embryonic cell (diameter: 800 μm) is successfully conducted. The results of this experiment not only validate the precision and effectiveness of the proposed microrobot and its associated models but also highlight its tremendous potential for cell manipulation in vitro and in vivo.

Abstract Background Adolescent non-suicidal self-injury (NSSI) is common and adolescence is the most common period of first self-injury, and the occurrence of NSSI is influenced by negative life events and emotional symptoms. The mediating role of emotional symptoms in the interaction between negative life events and NSSI has not been carefully investigated yet. Methods For middle school students in three schools in a Chinese province, the Adolescents Self-Harm Scale was used to investigate NSSI, the Adolescent Self-Rating Life Events Check List was used to investigate adolescent negative life events, and the Self-Rating Anxiety Scale and Self-Rating Depression Scale were used to assess their emotional symptoms. After the description of general data and the test for differences between groups, the relationship between negative life events, emotional symptoms and NSSI was analyzed using Pearson correlation analysis. Structural equation modeling was used to analyze the mediating role of emotions in negative life events and NSSI. Results A total of 2376 junior high school students completed this survey, which revealed an annual NSSI prevalence of 37.1% (n = 881) and a higher prevalence of NSSI among girls and rural adolescents. Among adolescents who developed NSSI, 67.4% (N = 594) used multiple means of self-injury. The most common means of self-injury was hair pulling (51.0%), and the most common NSSI purpose and external factors/events were venting bad emotions or feelings (57.5%) and poor academic performance (44.9%), respectively. Negative life events, emotional symptoms and NSSI were positively associated (P

Endometriosis is one of the most common benign gynecological disorders in reproductive-aged women. The major symptoms are chronic pelvic pain and infertility. Despite its profound impact on women’s health and quality of life, its pathogenesis has not been fully elucidated, it cannot be cured and the long-term use of drugs yields severe side effects and hinders fertility. This review aims to present the advances in pathogenesis and the newly reported lead compounds and drugs managing endometriosis. This paper investigated Genetic changes, estrogen-dependent inflammation induction, progesterone resistance, imbalance in proliferation and apoptosis, angiogenesis, lymphangiogenesis and neurogenesis, and tissue remodeling in its pathogenesis; and explored the pharmacological mechanisms, constitutive relationships, and application prospects of each compound in the text. To date, Resveratrol, Bay1316957, and bardoxifene were effective against lesions and pain in controlled animal studies. In clinical trials, Quinagolide showed no statistical difference with the placebo group; the results of phase II clinical trial of the IL-33 antibody have not been announced yet; clinical trial stage III of vilaprisan was suspended due to drug toxicity. Elagolix was approved for the treatment of endometriosis-related pain, but clinical studies of Elagolix for the pretreatment of patients with endometriosis to before In vitro fertilization treatment have not been fulfilled. The results of a clinical study of Linzagolix in patients with moderate to severe endometriosis-related pain have not been disclosed yet. Letrozole improved the fertility of patients with mild endometriosis. For endometriosis patients with infertility, oral GnRH antagonists and aromatase inhibitors are promising drugs, especially Elagolix and Letrozole.

With the advance in algorithms, deep reinforcement learning (DRL) offers solutions to trajectory planning under uncertain environments. Different from traditional trajectory planning which requires lots of effort to tackle complicated high-dimensional problems, the recently proposed DRL enables the robot manipulator to autonomously learn and discover optimal trajectory planning by interacting with the environment. In this article, we present state-of-the-art DRL-based collision-avoidance trajectory planning for uncertain environments such as a safe human coexistent environment. Since the robot manipulator operates in high dimensional continuous state-action spaces, model-free, policy gradient-based soft actor-critic (SAC), and deep deterministic policy gradient (DDPG) framework are adapted to our scenario for comparison. In order to assess our proposal, we simulate a 7-DOF Panda (Franka Emika) robot manipulator in the PyBullet physics engine and then evaluate its trajectory planning with reward, loss, safe rate, and accuracy. Finally, our final report shows the effectiveness of state-of-the-art DRL algorithms for trajectory planning under uncertain environments with zero collision after 5,000 episodes of training.

Abstract Small interfering RNA (siRNA)-based gene therapy has provided an alternative strategy for cancer therapy. One of the key components within gene therapy process is the delivery system. As a novel non-viral gene vector, DMP, prepared by modifying mPEG-PCL micelle with cationic DOTAP lipid, has been prepared and successfully applied in plasmid DNA-based colon cancer gene therapy study. However, its potential in siRNA delivery is unknown. In this study, the preparation process of DMP was optimized and the anti-cancer efficacies of the DMP/siMcl1 and DMP/siBcl-xl complexes were studied on a mouse colon cancer model. Our results demonstrated that DMP cationic micelle-delivered siRNAs could effectively inhibit the growth of C26 colon cancer cells in vitro. Meanwhile, intratumoral administration of DMP/siMcl1 and DMP/siBcl-xl complexes obviously suppressed subcutaneous tumor model in vivo. These results suggest the DMP/siRNA complex to be a potential candidate for cancer gene therapy.

The decompressive laminectomy is a common treatment for lumbar spinal stenosis. Generally, surgeons use grinding tools to remove laminae under the guidance of intraoperative medical images. To improve accuracy and reduce surgeons' burdens, robot-assisted surgery is gaining acceptance. This paper proposes a method to plan grinding paths and velocities based on 3-D medical images in the context of robot-assisted decompressive laminectomies. As the lesion areas to be grinded are irregular, an interactive method with 3-D reconstruction is designed for surgeons to transfer discrete information about grinding paths and velocities to the robot system. The path generation strategy is based on a ray casting algorithm after space registrations, while the velocity generation strategy is based on the virtual force and mechanical analysis is used to optimize temporal efficiency and stability. A complete system is developed to test and explore the feasibility of this method. Results suggest that robot-assisted decompressive laminectomies can be performed well.

DNA damage and base excision repair (BER) are actively involved in the modulation of DNA methylation and demethylation. However, the underlying molecular mechanisms remain unclear. In this study, we seek to understand the mechanisms by exploring the effects of oxidative DNA damage on the DNA methylation pattern of the tumor suppressor breast cancer 1 (BRCA1) gene in the human embryonic kidney (HEK) HEK293H cells. We found that oxidative DNA damage simultaneously induced DNA demethylation and generation of new methylation sites at the CpGs located at the promoter and transcribed regions of the gene ranging from −189 to +27 in human cells. We demonstrated that DNA damage-induced demethylation was mediated by nucleotide misincorporation by DNA polymerase β (pol β). Surprisingly, we found that the generation of new DNA methylation sites was mediated by coordination between pol β and the de novo DNA methyltransferase, DNA methyltransferase 3b (DNMT3b), through the interaction between the two enzymes in the promoter and encoding regions of the BRCA1 gene. Our study provides the first evidence that oxidative DNA damage can cause dynamic changes in DNA methylation in the BRCA1 gene through the crosstalk between BER and de novo DNA methylation.

Dysfunctional stem cell differentiation into placental lineages is associated with gestational diseases. Of the differentiated lineages available to trophoblast stem cells (TSC), elevated O2 and mitochondrial function are necessary to placental lineages at the maternal–placental surface and important in the etiology of preeclampsia. TSC lineage imbalance leads to embryonic failure during uterine implantation. Stress at implantation exacerbates stem cell depletion by decreasing proliferation and increasing differentiation. In an implantation site O2 is normally ~2%. In culture, exposure to 2% O2 and fibroblast growth factor 4 (FGF4) enabled the highest mouse TSC multipotency and proliferation. In contrast, hypoxic stress (0.5% O2) initiated the most TSC differentiation after 24 h despite exposure to FGF4. However, hypoxic stress supported differentiation poorly after 4–7 days, despite FGF4 removal. At all tested O2 levels, FGF4 maintained Warburg metabolism; mitochondrial inactivity and aerobic glycolysis. However, hypoxic stress suppressed mitochondrial membrane potential and maintained low mitochondrial cytochrome c oxidase (oxidative phosphorylation/OxPhos), and high pyruvate kinase M2 (glycolysis) despite FGF4 removal. Inhibiting OxPhos inhibited optimum differentiation at 20% O2. Moreover, adding differentiation-inducing hyperosmolar stress failed to induce differentiation during hypoxia. Thus, differentiation depended on OxPhos at 20% O2; hypoxic and hyperosmolar stresses did not induce differentiation at 0.5% O2. Hypoxia-limited differentiation and mitochondrial inhibition and activation suggest that differentiation into two lineages of the labyrinthine placenta requires O2 > 0.5–2% and mitochondrial function. Stress-activated protein kinase increases an early lineage and suppresses later lineages in proportion to the deviation from optimal O2 for multipotency, thus it is the first enzyme reported to prioritize differentiation.

Female animals living in the Qinghai-Tibet Plateau have lower ovulation rates because of the hypoxic environment, however, the mechanism of hypoxia on animal follicles is unclear. In this study, the effects of hypoxia on bovine follicles were investigated using an in vitro follicular culture system. The results show that there was a significant decrease in follicular diameter from day 3 to day 6 in both hypoxia and hypoxia with estrogen (E2) and fulvestrant (ICI 182780) (hypoxia + E2 + ICI) groups, when compared with a normoxia group (p < 0.05). We also observed significant downregulation of ERα and FSHR, while upregulation of LHCGR expression in the hypoxia group and hypoxia + E2 + ICI groups compared to the normoxia group (p < 0.05). The expression of IGF1 gene was significantly downregulated in hypoxia + E2 + ICI group when compared to the hypoxia + E2 group (p < 0.05). The expression of HIF1A, ADAMTS1, VEGFA, and EDN2 were upregulated in both hypoxia and hypoxia + E2 + ICI groups in comparison to normoxia group (p < 0.05). Under hypoxic conditions, the addition of E2 resulted in a decrease of HIF1A protein but an increase of ERα protein in cultured bovine follicles (p < 0.05). In summary, hypoxia limits the growth of bovine follicle cultured in vitro through inhibition of ERα.

In the present study, AA was used to challenge bovine ovarian granulosa cells in vitro and the related parameters of cellular and molecular biology were measured. The results indicated that lower doses of AA increased survival of bovine granulosa cells whereas higher doses of AA suppressed survival. While lower doses of AA induced accumulation of lipid droplet in granulosa cells, the higher dose of AA inhibited lipid accumulation, and AA increased abundance of FABP3, CD36 and SLC27A1 mRNA. Higher doses of AA decreased the secretion of E2 and increased the secretion of P4 accompanied by down-regulation of the mRNA abundance of CYP19A1, FSHR, HSD3B1 and STAR in granulosa cells. The signaling pathways employed by AA in the stimulation of genes expression included both ERK1/2 and Akt. Together, AA specifically affects physiological features, gene expression levels and steroid hormone secretion, and thus altering the functionality of granulosa cells of cattle.