EGFR+ non‐small cell lung cancer (NSCLC) patients with leptomeningeal metastasis (LM) comparing survival rates and gene mutation detection with matched cerebrospinal fluid (CSF) and plasma are relatively scarce. We evaluated gene mutations, treatment strategies, and clinical outcomes in EGFR+ NSCLC patients with LM.Studies in EGFR+ NSCLC patients with LM for prognostic significance.We retrospectively reviewed gene mutation status in the CSF and plasma of 32 P = 0.000). Overall survival was 27.0 months , median survival after LM was 7.0 months (95% CI 5.0–11.0), and median survival before LM was 17.0 months (95% CI 12–25.5). Median survival after LM was significantly shorter in patients with “worse” status (n = 7) than in those with “improved/stable” status .The rate of LM disease control was significantly higher in patients that switched EGFR‐tyrosine kinase inhibitor (TKI) treatments, initiated EGFR‐TKIs, or received high‐dose EGFR‐TKI treatment than those who continued their current EGFR‐TKI treatment, received chemotherapy, or were not administered antitumor treatment (24/24, 100.0% vs. 1/8, 12.5%; EGFR+ NSCLC patients after a diagnosis of LM. Liquid biopsy in both plasma and CSF, as well as dynamic detection, play important roles in the direction of treatment for such patients.EGFR‐TKIs should be the priority course of treatment in  EGFR+ NSCLC patients, and 9.0% of EGFR+ NSCLC patients were diagnosed with LM.Several kinds of solid tumors, including malignant melanoma and lung, breast, and gastric cancer are accompanied by leptomeningeal metastasis (LM) at some stage during disease progression. Among these cancers, non‐small cell lung cancer (NSCLC) accounts for approximately 40–50% of cases that include LM.EGFR gene mutations (EGFR+) tend to have more central nervous system (CNS) metastases, including both brain and leptomeningeal metastases after using of EGFR‐tyrosine kinase inhibitors (TKIs) in association with theuse of EGFR‐tyrosine kinase inhibitors (TKIs). There are several potential reasons for this phenomenon. Firstly, the survival of EGFR+ NSCLC patients has been significantly prolonged.EGFR+ NSCLC patients with LM generally include an intial high‐dose of first or second‐generation EGFR‐TKIs, or a switch to a different EGFR‐TKI.An increasing number of reports have shown that NSCLC patients with EGFR mutations can be detected by analyzing circulating tumor DNA (ctDNA) in both plasma and cerebrospinal fluid (CSF), which provides an alternative to tumor biopsies for further study. With the widespread use of third generation EGFR‐TKIs, data on LM in NSCLC patients with EGFR+ is relatively scarce. We retrospectively assessed the diagnosis, treatment modes, and survival status of EGFR+ NSCLC patients with LM that had been enrolled in our hospital. We determined their gene mutation status by analyzing ctDNA extracted from peripheral blood, CSF, and tumor tissue samples.EGFR+ NSCLC patients diagnosed with advanced‐stage (stage IV) NSCLC between January 2016 and November 2018 were reviewed for diagnosis of LM. All LM patients needed to be diagnosed by magnetic resonance imaging (MRI) and/or by the detection of malignant cells in the CSF by cytopathological diagnosis. In the event that only atypical and/or suspicious cells were found in the CSF, patients were not diagnosed with LM.In this retrospective study, All patients signed informed consent to participate in this study and gave permission to use their peripheral blood and CSF. The ethical committee of the Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College approved the protocol.EGFR mutation status of patients in the following regions: EGFR driver mutations , EGFR resistance mutations , or mutations in tumor suppressor genes of plasma and CSF ctDNA. EGFR mutation status was determined for all patients by analyzing ctDNA exracted from tumor samples and/or CSF. Paired CSF and plasma samples were collected at the same time from the majority of patients. Approximately 10 mL of CSF was collected by lumbar puncture for cytology examination and gene detection by amplified refractory mutation system (ARMS) or next‐generation sequencing (NGS). Additionally, 8 mL of plasma was collected for super AMRS or NGS analysis concurrently with the collection of CSF.We ascertained the Follow‐up continued until January 2019. The duration of investigation was calculated from the initial diagnosis of LM until death or the last date of follow‐up, with a minimum follow‐up period for statistical analysis set at one month. The primary outcome measurement was disease control of LM, which was evaluated using the following criteria: improved/stable, defined as clinical symptom improvement or maintaining a consistent state, and/or enhanced computed tomography (CT) or MRI examination showing decreased or stable lesion state; worse, defined as a worsening of clinical symptoms or observation of increased lesions on enhanced CT or MRI examination according to European Association of Neuro‐Oncology–European Society for Medical Oncology (EANO–ESMO) Clinical Practice Guidelines.2 test and independent t‐test, respectively. Survival analyses were performed according to the Kaplan–Meier method and tested for significance with the log‐rank test. Comparisons between subgroups were made using a Cox proportional hazards model and Wald 95% confidence intervals (CIs), where appropriate. Statistical analyses were performed using SPSS version 19.Comparisons of categorical and continuous variables were performed using the χEGFR+ advanced‐stage NSCLC with LM were reviewed. LM was diagnosed by both MRI and CSF cytology in 20 patients (62.5%), only by CSF cytology in 10 patients (31.3%), and only by MRI in two patients (6.3%) , and there were significant differences in duration between patients diagnosed within 12 and over 12 months. . A total of 28 patients had brain metastasis (BM) before LM or at the same time as LM diagnosis . Eastern Cooperative Oncology Group performance status (ECOG PS) was evaluated at the time of LM diagnosis. The ECOG PS of 28 patients was 0–2 , and 25 patients were assessed with SD/PR of extracranial tumors   had exon 19 deletions, 15 (51.7%) had exon 21 L858R, 1 (3.4%) had an exon 20 insertion, and 1 patient (3.4%) had an ERBB2 alteration  underwent EGFR driver mutation detection analysis on ctDNA extracted from the CSF after LM diagnosis: 20 (62.5%) were tested by ARMS, and 12 were tested by NGS. The results revealed: 13 patients (37.5%) had exon 21 L858R mutations; 8 (25.0%) had exon 19 deletions; 6 (18.8%) had wild‐type EGFR; 1 (3.1%) had a T790M mutation; 1 (3.1%) had an exon 19 deletion and T790M mutation; 1 (3.1%) had an exon 20‐C797S mutation and exon 19 deletion; 1 (3.1%) had an exon 20‐770 alteration; and 1 patient (3.1%) had an exon 18 alteration  had T790M mutations, 6 (50.0%) had TP53 alterations, and 7 (58.3%) had EGFR mutations via ARMS versus NGS in CSF showed different results. The detection rate of EGFR T790M and exon 20 mutations was significantly higher by NGS than by ARMS .Analysis of EGFR; 4  had EGFR exon 19 deletions; 3  had EGFR exon 21 L858R mutations; 2  had HER2 alterations; 2  had EGFR T790M mutations; 1  had ROS‐1 alterations; and 1 patient  had EGFR exon 21 L858R and T790M mutations  had wild‐type EGFR driver mutations and ROS‐1 in plasma, while 24 patients showed EGFR driver mutations in CSF . No mutations were detected in the plasma and CSF of 16 and 6 patients, respectively   (Table P = 0.474)  were treated using a specfic EGFR‐TKI. Five different regimens were used during follow‐up treatment for LM according to the personal and specific conditions of each patient: EGFR‐TKIs, chemotherapy, WBRT, ITCT, and TMZ. EGFR‐TKI treatment included: continuation of current EGFR‐TKI treatment, initiation of EGFR‐TKI treatment, switching from one EGFR‐TKI treatment to another, and high‐dose EGFR‐TKI treatment Table . There w0) Table . Among t.4% were 4) Table .EGFR driver and/or T790M‐positive mutations and those without EGFR driver mutations or T790M‐negative  (Table There was a significant difference in the rate of LM disease control between patients with 7) Table .EGFR+ NSCLC patients with LM died before their follow‐up appointments and the one‐year survival rate was 59.4% (19/32). The OS of all 32 patients was 27.0 months (95% CI 19.0–37.5)  of the 2, 21.9% EGFR driver and/or T790M positive mutations in both plasma and CSF; 20.5 months (95% CI 12.0–48.0) in 14 patients with EGFR driver mutations and/or T790M negative in plasma but positive in CSF; 20.0 months (95% CI 6.0–30.0) in 3 patients with EGFR driver and/or T790M positive mutations in plasma but negative in CSF; and 18.0 months (95% CI 0.0–27.0) in 3 patients with EGFR driver mutations and T790M negative in both plasma and CSF. The median survival duration in 17 patients with EGFR driver mutations and/or T790M negative in plasma was longer than in 12 patients with EGFR driver and/or T790M positive mutations in plasma   : 15.0 months (95% CI 4.0–27.0) in 9 patients with EGFR driver and/or T790M positive mutations in both plasma and CSF; 7.2 months (95% CI 4.1–11.0) in 14 patients with EGFR driver mutations and/or T790M negative in plasma but positive in CSF; 4.5 months (95% CI 1.5–10.0) in 3 patients with EGFR driver and/or T790M positive mutations in plasma but negative in CSF; and 14.0 months  in 3 patients with EGFR driver mutations and T790M negative in both plasma and CSF.The median survival duration after LM of all 32 patients was 7.0 months (95% CI 5.0–11.0): 11.4 months (95% CI 5.5–20.6) in 9 patients with n = 25), the median survival duration after LM was significantly shorter in patients with worse LM  . Because EGFR+ patients had higher ECOG PS scores at the time of LM diagnosis, patients with unspecific symptoms, such as headaches and dizziness, need to have their LM status taken into consideration.In previous studies, researchers reported that NSCLC patients not administered systemic chemotherapy, chemotherapy, or EGFR‐TKI treatment as a basic option after LM diagnosis only survived three months.EGFR+ NSCLC patients with LM has produced various results.et al. reported that 12 EGFR+ NSCLC patients with LM treated with high dose first‐generation EGFR‐TKIs after LM diagnosis achieved survival after LM of only 2.4 months.Because of the BBB, the concentrations of available first and second‐generation EGFR‐TKIs in CSF, such as gefitinib, erlotinib, and afatinib, are considerably lower compared to the concentration in extracranial lesions.Afatinib is a second generation EGFR‐TKI. A recent study showed that if patients experienced disease progression after treatment of standard doses of erlotinib or gefitinib, 66% of them could achieve disease control when they switched to afatinib.n = 16), whereas all 11 patients not administered ITCT achieved disease control, which could be associated with the small sample size. Third generation EGFR‐TKIs combined with ITCT did not achieve a higher rate of LM disease control compared to third genenration EGFR‐TKIs alone. Unfortunately, there are no satisfactory survival results available related to ITCT.In recent years, ITCT has been a traditional treatment choice for NSCLC patients with LM, but evidence of its efficacy is limited. In a sample of 105 NSCLC patients with LM, ITCT use resulted in OS of only 3.0 months. Furthermore, varying symptoms, such as headaches, altered mental state, and cauda equine, showed relatively low response rates, ranging from 42% to < 10%, and negative conversion of CSF cytology could not prolong OS. In our study, only 16 out of 21 patients administered ITCT achieved disease control  with gene mutations in common. In 12 patients diagnosed by NGS, more types of gene mutations in CSF were associated with both secondary resistance genes and suppressors, in addition to other EGFR driver mutations. Therefore, detecting genomic alterations in CSF directly, especially by NGS, is a feasible method to represent the genetic status of intercranial lesions. Similarly, ctDNA in plasma provides an alternative to tumor samples for EGFR mutation analysis, and plasma gene mutations showed greater advantages than solid tumor cells for revealing the genetic landscape of primary and metastatic lesions, which are relatively limited. In the BENEFIT clinical trial, EGFR mutations reappeared in ctDNA with plasma samples when the disease progressed in 46% (56/123) of patients.EGFR driver mutations and ROS‐1 in plasma when LM appeared, which is relatively low compared to the results of the BENEFIT analysis. However, in our study, more patients had EGFR driver mutations in CSF and more without EGFR driver mutations in plasma when LM was diagnosed, which is relatively high compared to the BENEFIT results. Therefore, LM might have a different mechanism from extracranial progression, and further studies need to be conducted to verify this possibility. Gene mutation status in an individual patient can be spatiotemporally heterogeneous, and in this case, it is necessary to detect multiple lesions simultaneously. The median survival duration before LM of 17 patients with EGFR driver mutations and/or T790M negative in plasma was 18.0 months, which was longer than in patients found to be T790M mutation positive in plasma. Subgroup analysis indicated that the median survival before LM of patients with EGFR driver mutations and/or T790M mutation negative in plasma and administered first‐generation EGFR‐TKIs was 24.0 months, which was longer than that when found positive in plasma. This result suggests that patients without gene mutations in plasma could have longer survival before LM, although the difference between subgroups was not significant. A similar result was observed in the BENEFIT study.In our study, SD or PR was achieved in more extracranial tumors than in a previous study of unselected NSCLC patients with LM which reported that PD was associated with extracranial tumors. It is possible that extracranial tumors and LM arise from different mechanisms.EGFR driver mutations and/or T790M was significantly higher than in patients  without EGFR driver mutations and T790M. However, acquired resistance to osimertinib was also observed, and the molecular mechanisms of resistance  are not yet fully understood. In our study, one patient developed an EGFR‐C797S resistance mutation in CSF accompanied with an EGFR exon 19 deletion  after osimertinib treatment. Therefore, NGS with more advanced, targeted, ultra‐deep sequencing in CSF, which could more effectively monitor the gene mutation status and provide clinicians more opportunities to adjust EGFR‐TKI therapy for patients after acquired resistance to classic EGFR‐TKIs is needed.Acquired resistance to EGFR‐TKIs is a common clinical problem, and the T790M mutation has been confirmed by several studies to be one of the most important biomarkers of acquired resistance.TP53 is a common tumor‐suppressor gene and is more highly expressed in patients whose disease transforms to SCLC (82%), which is another factor for EGFR‐TKI resistance in lung adenocarcinoma.TP53  was uniquely identified in CSF ctDNA, but there was no significant difference in disease control between patients with or without TP53. This result suggests that TP53 may play a role in the progression of LM, but larger sample size studies are required to further explore this possibility.ALK, ROS‐1, EGFR exon 20 insertions, and BRAF in the CSF. After taking alectinib for one month, the LM symptoms in the CNS improved. Another patient was found to have L858R in both CSF and plasma when diagnosed with LM, and achieved disease control for 14 months until clinical deterioration in CNS was observed. Re‐biopsy revealed EGFR L858R and MET alterations in the CSF. Another patient with EGFR exon 19 deletion in plasma and EGFR exon 19 deletion and exon 20 insertion‐C797S in CSF when diagnosed with LM achieved disease control for seven months until the appearance of clinical deterioration in CNS. Re‐biopsy detected coexisting EGFR exon 19 deletion, EGFR exon 20 insertion‐C797S, MET, BRAF, KRAS, and EGFRamp in the CSF. These results collectively suggest that it is important to re‐biopsy in clinical practice after disease progression in order to find any new mutations and alter therapeutic strategies.In our study, three patients had re‐biopsies after presenting with clinical deterioration in the CNS. One patient was administered eight ITCT treatments and systemic treatment for five months, and after clinical deterioration in the CNS was observed, re‐biopsy detected alterations of EGFR mutations detected by other methods, such as NGS, and concentrate on dynamic detection.Our study has several limitations. First, the study was retrospective with a small sample size, particularly regarding the use of NGS. Second, the rate of dynamic monitoring of ctDNA in CSF and plasma during treatment after LM was low. We plan to investigate the efficacy of EGFR‐TKIs in prospective trials based on EGFR+ NSCLC patients with LM. The median survival duration after diagnosis of LM was 7.0 months. EGFR‐TKIs should be considered priority treatment, as they are associated with superior survival in NSCLC patients after a diagnosis of LM. Tissue and liquid biopsies from multiple sites, as well as dynamic detection by NGS, play important roles in determining treatment strategies for EGFR+ NSCLC patients with LM.In conclusion, we describe a cohort of No authors report any conflict of interest.


Fos, Arc, and Bdnf) induced by SD, suggesting intact synaptic upscaling on the transcript level during extended wakefulness with aging. However, sleep‐activated processes, such as DNA repair, synaptogenesis, and axon guidance, were sensitive to the effect of aging. Old mice expressed elevated levels of immune response genes when compared to young mice, and enrichment analysis using cell‐type‐specific markers indicated upregulation of microglia and oligodendrocyte genes in old mice. Moreover, gene sets of the two cell types showed age‐specific sleep/wake regulation. Ultimately, this study enhances understanding of the transcriptional changes with sleep and aging, providing potential molecular targets for future studies of age‐related sleep abnormalities and neurological disorders.Sleep abnormalities are common with aging. Studies show that sleep plays important roles in brain functions, and loss of sleep is associated with increased risks for neurological diseases. Here, we used RNA sequencing to explore effects of age on transcriptome changes between sleep and sleep deprivation (SD) in medial prefrontal cortex and found that transcriptional changes with sleep are attenuated in old. In particular, old mice showed a 30% reduction in the number of genes significantly altered between sleep/wake and, in general, had smaller magnitudes of changes in differentially expressed genes compared to young mice. Gene ontology analysis revealed differential age effects on certain pathways. Compared to young mice, many of the wake‐active functions were similarly induced by SD in old mice, whereas many of the sleep‐active pathways were attenuated in old mice. We found similar magnitude of changes in synaptic homeostasis genes ( More recently, gene expression has also been studied in specific cell types using laser capture microdissection , a brain region that likely plays a major role in age‐related changes in sleep/wake behavior and where beta‐amyloid burden has been correlated with reduced slow‐wave activity  analysis, samples from different behavioral states and ages form separate clusters on a three‐dimensional space. Sleep deprivation and spontaneous sleep (SS) samples of the same age are clearly separated on the first two dimensions Figure a, indica2.2We assessed differentially expressed genes (DEGs) between sleep deprived mice and time‐matched sleep controls at four time points. Using a false discovery rate (FDR) of 1%, we found 5,158 DEGs in young mice and only 3,540 DEGs in old mice. This includes 5,833 unique genes ; 2,865 (49.1%) were differentially expressed in both ages, 2,293 (39.3%) regulated only in young mice (young‐specific), and 675 (11.6%) regulated only in old mice (old‐specific)  cutoff of 0.2 (~15% change in expression) was applied to all DEGs to remove genes with small FCs (averaged over the time points). This resulted in 3,802 and 2,532 DEGs for young and old mice, respectively. Nearly equal numbers of genes were upregulated by SD (wake genes) or upregulated during SS (sleep genes)  Figure . Table S2.310p‐values compared to young mice, not only in young‐specific DEGs, but, more importantly, also in common DEGs  showed significant interaction at an FDR < 5% Figure .E130307A14Rik, Gm15513, Pclo, Reln, Slc22a3, Epb4.1l5, Herc1, and Meig1) and 1 common gene (Slit2). The remaining 3 genes were old‐specific . These 12 sleep genes are involved in various functions; the most overrepresented include positive regulation of peptidyl‐tyrosine phosphorylation , cell adhesion , and protein localization to synapse  , 4 are common , and 1 is old‐specific (Syne1). No biological function was enriched among these wake genes.Of the 12 sleep genes, 9 had greater changes in young mice, including 8 young‐specific genes (2) Table . Of the 2.5p < .05).As interaction tests are underpowered, we further examined age differences in the common and age‐specific DEGs at the pathway level using functional clustering analyses. Eleven functions were enriched among sleep genes, including three among the common DEGs, five among the young‐specific DEGs and three among the old‐specific DEGs. Table 2FC of the genes involved in each enriched function was compared between young and old mice. Young animals showed greater increases in expression during SS [compared to both SD  as well as all five young‐specific functions . Medium to large expression differences (effect sizes [Cohen's d] between 0.42 and 2.04) were found between young and old mice for all pathways except Signal/transmembrane . Interestingly, expression differences in the old‐specific pathways appear driven by greater decrease in SD , rather than increase in SS  showed significantly greater expression increase in SD in young mice . Similarly, functions enriched among old‐specific DEGs showed either significant  or moderate  increase in SD in old mice. However, unlike the common sleep functions, all 12 common wake functions showed small differences between young and old mice (Cohen's d ≤ 0.31). While effects were small, the four functions with most genes (n > 50) showed highly significantly increased expression changes in young mice compared to old   Table .2.72FC cutoff , 536 DEGs during sleep and 252 DEGs at ZT0 remained. More genes were found upregulated with age (341 during SS and 144 at ZT0) than downregulated with age (195 during SS and 108 at ZT0)  Figure . Table Santigen processing/presentation, inflammatory response, and immune system process. The homophilic cell adhesion pathway is also induced both during sleep and at ZT0 in old mice. During sleep, a few additional pathways were enriched among age upregulated genes, including lipid metabolic process, positive regulation of myelination, apoptotic process, and negative regulation of neuron development. See Table Multiple immune pathways are enriched from genes upregulated both during SS and at ZT0 in old mice, including signal transduction, locomotory behavior, chemical synaptic transmission, and nervous system development are the top enriched functions  were affected by sleep/wake and aging.Since circadian‐related functions were enriched by both behavior and age, we specifically examined how the 12 circadian core clock genes  and nonsignificant effect for Per3  were significantly altered by SD in old mice. With respect to the effects of age, Cry1 was highly elevated in old mice during sleep (q = 9.3e−5). Arntl, Per1, Per2, and Per3 also showed moderate changes between young and old mice at ZT0 or during SS (q = 0.01–0.05). Cry1, Per1 and Per2 showed reduced change during SS relative to ZT0 in old compared to young mice , except for a trending effect in r3 Table . In compe Figure , suggest2.9We found multiple synaptic transmission‐ and synaptic plasticity‐related pathways regulated by behavior or age. Thus, we examined how known genes involved in synaptic homeostasis and synaptogenesis were affected in our data Table .Fos, Fosb, Egr1, Egr2, and Arc. All were highly induced by SD in both ages (q ≤ 6.4e−17), with similar magnitude of change in young (2.6‐ to 7.7‐fold increase) and old (2.8‐ to 11.5‐fold increase) mice. Bdnf and its receptor Ntrk2 (or TrkB) were also upregulated by SD in both young and old mice (Table As many transcription factors (TFs) differentially regulated by sleep and wake are key factors of neuronal activity and synaptic plasticity , while changes were reduced or nonsignificant in old mice  were not differentially regulated, as indicated in previous studies  were significantly induced during SS compared to SD in young mice, and changes in old mice were either reduced or abolished . Also, downregulation of oligodendrocyte precursor cells (OPCs) and neuron marker genes were observed in the old mice during sleep.Comparing young and old mice Figure a, microgComparing sleep and wake Figure b, upregu33.1This is the first RNA‐Seq study comparing sleep/wake transcriptomic regulations between young and old mice in mPFC. When examining expression differences between SD and SS at four time points over the lights‐on period (ZT0–ZT12), we identified 5,158 DEGs in young mice and 3,540 DEGs in old mice at an FDR of 1%. Compared to our prior microarray study of cortex of young mice  or SS (sleep genes). Thus, there is a 33% reduction in the number of sleep and wake DEGs in the old mice. This reduction in the number of DEGs is driven by a reduction in the magnitude of expression changes between sleep and wake in old mice, rather than increased variability with aging, including in ~70% of common DEGs differentially regulated in both ages.ER stress, regulation of transcription, and MAPK signaling, than sleep‐activated functions, such as DNA repair. Also, many more sleep‐activated genes/functions, such as synaptic transmission, synaptogenesis‐related cell adhesion, axon guidance, and neurogenesis, were no longer differentially regulated in the old mice. Thus, transcriptional changes during sleep are particularly attenuated in old mice.While we found an overall reduction in expression changes between sleep and wake in old mice, not all pathways showed age‐related effects. Thus, it cannot simply be ascribed to alterations in the nature of sleep. In general, more wake‐activated functions were commonly enriched in young and old mice, such as 3.2Fos, Arc, and Bdnf) in both ages at nearly identical magnitudes. We also did not observe any expression difference of Arc and Fos between young and old mice during undisturbed sleep. Some studies observe age‐related reduction of Arc and Fos or response of Fos to stress in select brain regions or cell types , known regulators of NMDARs and neurotransmitter release , synapsin (Syn1 and Syn2), and neurexin (Nrxn1 and Nrxn3), important players in presynaptic assembly , an essential gene for neuronal development   were upregulated with sleep in young and old mice. Greater changes during sleep were generally observed in old mice. Moreover, we identified enrichment of lipid metabolic process and positive regulation of myelination among upregulated sleep genes in old mice. Oligodendrocytes marker genes were also significantly increased in old versus young mice during sleep. These results are consistent with prior studies. For example, studies have found age‐related increases in oligodendrogenesis and myelinogenesis in the spinal cord of mice  were induced by sleep in both ages, as seen previously in cortex of young mice , an essential factor for protein synthesis   protein is not present in aged mice  involved in ER stress in old mice, suggesting a subtle attenuation at the pathway level with aging.Sleep deprivation‐induced ER stress is shown in different brain regions and across multiple species  are specific markers of the “primed” microglia, a status associated with aging and neurodegenerative diseases  and MHC class II genes (H2‐Q6 and H2‐Q7) in old mice. As several studies link microglia activation with neurogenesis inhibition and synapse pruning  were downregulated during sleep only in young mice; thus, expression of these genes was elevated in old mice during sleep compared to young mice, suggesting potential roles of sleep/wake regulation in oligodendrocyte‐related neuronal protection.Besides microglia, we observed a positive enrichment of oligodendrocyte genes in old mice. Studies have shown an important cross talk between microglia and oligodendrocytes , again suggesting the molecular homeostatic response to forced wakefulness is not affected by aging. Results are supported by a recent study revealing unchanged expression of isolated astrocyte homeostasis genes with aging  were significantly different between SS and SD in young mice. In the old mice, only five of these genes were significantly regulated, suggesting a possible age‐related change on the effect of sleep/wake on circadian gene expression. A second limitation is SD via gentle handling can potentially introduce mild stress in addition to wakefulness. However, earlier study on rat Cirelli,  found si3.10Using RNA sequencing in mPFC, we identified a large number of differentially regulated genes impacted by sleep and wake in young and old mice. There was a system‐wide reduction in the number and magnitude of expression changes of the transcript level in old mice, suggesting an age‐associated attenuation in their sleep/wake regulation. Specifically, our study revealed that sleep‐activated processes  were sensitive to the effect of aging. The magnitude of change was attenuated in old mice. These results provide potential molecular targets for studying the connections between age‐related sleep disorders and neurodegenerative diseases.4Additional details and references are presented in the 4.1Experiments were performed on male C57BL/6 mice at 2–4 months (young) and 18–20 months (old) housed individually in a 12/12 light/dark room. Sleep deprivation was initiated with gentle handling at lights‐on  and continued for 3, 6, 9, and 12 hr in different groups. Time‐matched SS mice with ≥75% sleep in both ages in the last hour before sacrifice were collected. Randomly selected mice were sacrificed at ZT0 for baseline data. Six mice were collected in each condition.4.22 counts per million (CPM) > −1.4] and were normalized using the “Trimmed Mean of M‐values” (TMM) method.RNA extraction was performed from microdissected mPFC using an RNAqueous‐Micro Kit. Library preparation was performed using Illumina TruSeq Stranded mRNA kit and sequencing done with 100‐base pair single‐end reads on Illumina HiSeq 4,000. Raw reads were aligned to the mouse genome build mm9 by STAR version 2.5.3a, and quantified at the gene level using scripts from the PORT pipeline . A total of 14,656 genes with unique ensemble IDs passed filtering [mean log4.3F test was used to evaluate an ANOVA‐like null hypothesis of any pairwise differences between conditions (or ages) within each of the four separate time points. False discovery rate was controlled at 1% using the Benjamini–Hochberg method. To test if age modified the differences between SS and SD at any of the time points, we used statistical interaction tests; given lower power for interactions, an FDR threshold of 5% was used. Unless otherwise noted, FDR values and FCs between young and old mice were compared using Wilcoxon signed‐rank tests. DAVID was used for gene ontology (GO) enrichment analyses and functional clustering. Gene set enrichment tests were performed using CAMERA with Limma. Cell‐type‐specific marker gene sets  using LimmaVoom. For each gene, a moderated ts Table  were obtNone declared.A.I.P. designed the study and was involved in writing and editing. X.G. contributed to data collection, analysis, writing, and editing. B.T.K. contributed to data analysis, writing, and editing. D.S. and G.R.G. contributed to data analysis. D.C.L. contributed to data collection and editing. J.L. contributed to data collection. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file.


MDDGC9) is the rarest type of autosomal recessive muscular dystrophies. MDDGC9 is manifested with an early onset in childhood. Patients with MDDGC9 usually identified with defective glycosylation of DAG1, hence it is known as “dystroglycanopathies”. Here, we report a Chinese pedigree presented with mild MDDGC9. The proband is a 64 years old Chinese man. In this family, both the proband and proband's younger brother have been suffering from mild and late onset MDDGC9. Muscle biopsy showed that the left deltoid muscle with an advanced stage of dystrophic change. Immunohistochemistry staining of dystrophin, α‐sarcoglycan, β‐sarcoglycan and dysferlin are normal. Molecular genetic analysis of the proband has been done with whole exome sequencing. A homozygous novel missense mutation  in the exon 3 of the DAG1 gene has been identified in the proband. Sanger sequencing revealed that this missense mutation is co‐segregated well among the affected and unaffected (carrier) family members. This mutation is not detected in 200 normal healthy control individuals. This novel homozygous missense mutation (c.2326C>T) causes substitution of arginine by cystine at the position of 776 (p.R776C) which is evolutionarily highly conserved. Immunoblotting studies revealed that a significant reduction of α‐dystroglycan expression in the muscle tissue. The novelty of our study is that it is a first report of DAG1 associated muscular dystrophy‐dystroglycanopathy (limb‐girdle), type C, 9 (MDDGC9) with mild and late age of onset. In Chinese population this is the first report of DAG1 associated MDDGC9.Muscular dystrophy‐dystroglycanopathy (limb‐girdle), type C, 9 ( DAG1 gene encodes dystroglycan protein which is the key component of DGC and plays a significant role in linking dystrophin with other proteins of ECM to form and give mechanical support to the intracellular cytoskeleton. DAG1 gene is evolutionarily highly conserved and dystroglycan is primarily expressed as a precursor protein. During post‐translational modification, dystroglycan protein has been cleaved at Ser654 by proteolysis to form α‐dystroglycan and β‐dystroglycan.The dystroglycan  with mild autosomal recessive MDDGC9 was enrolled in the Department of Neurology, Peking Union Medical College Hospital, Beijing, China Figure . The stu2.2We performed comprehensive clinical examinations related to neuromuscular disorders. We explored routine blood tests including serum creatine kinase level. Electrophysiological study and muscle biopsy were performed. The muscle pathology procedures included standard staining and the immunohistochemistry staining of dystrophin‐N, dystrophin‐R, dystrophin‐C, α‐sarcoglycan, β‐sarcoglycan and dysferlin‐C .2.3The genomic DNA was extracted from peripheral blood, randomly fragmented and sheared into fragments of 180‐280 bp in length using a Covaris crusher. After the ends were repaired and A tails were added, DNA fragments were ligated to the ends of the fragments to prepare DNA libraries. The exome was enriched using Agilent's SureSelect Human AII Exon V5 Kit, with up to 543872 biotins after library pooling with a specific index. The labelled probes were hybridised in liquid phase. Then 334378 exons of 20965 genes were captured using streptomycin‐containing magnetic beads. The libraries were linearly amplified by PCR and subjected to library quality tests. After passing the test, they were carried out high‐throughput deep sequencing on the Illumina HighSeq 2500  was used to align the sequencing reads with the GRCh37.p10. Then the aligned reads were realigned and recalibrated by GATK Indel Realigner and the GATK Base Recalibrator respectively (broadinstitute.org/). GATK UnifiedGenotyper (broadinstitute.org/) was used for identifying the single‐nucleotide variants (SNV) and small insertions or deletions (indel). Finally, Consensus Coding Sequences Database (20130630) was used for variant annotation. According to the minor allele frequency with a cut‐off value of <0.05 in four databases , we filtered and selected the variants. Classification of variants  has been done according to the variant interpretation guidelines of American College of Medical Genetics and Genomics (ACMG).The detailed variant interpretation process is described in Figure 2.4NM_001177639 of DAG1 was used.Sanger sequencing has been performed for validating the variants identified by whole exome sequencing. The primers are as follows; F‐ 5′‐GCTGGCCATACGGACTCATCC‐3′, R‐ 5′‐GCCTAGCTTACTATAGGGCCG‐3′. The reference sequence 2.5Muscle sections were prepared and lysed by using lysis buffer and separated by 10% SDS‐PAGE followed by transfer of these gels to PVDF membrane. Skimmed milk (5%) was used for blocking the membrane and mouse monoclonal antibody was used to probe α‐dystroglycan. α‐actinin was used as a loading control. Goat antimouse IgG was used to detect the bound primary antibody and performed western blot.2.6http://bigd.big.ac.cn/gsub/submit/gsa/list]. Accession number: CRA000997.All data used for the analyses in this report are available through Genome Sequence Archive, BIG Data Center in Beijing Institute of Genomics (GEO) [URL: 33.1The proband is a 64 years old man. The proband was born from a consanguineous Chinese family Figure . The ons9, Neut% 59.6%, Ly% 31.8%, RBC 4.84 × 1012, Hb 150 g/L, PLT 287 × 109) and coagulation testing were normal. HBsAg, HIV‐Ab, HCV‐Ab and TP‐Ab were negative.The results of blood tests indicated serum muscle enzymes were slightly elevated (CK 1049‐1184U/L (24‐195), LDH 323 U/L↑(0‐250), AST 42U/L↑(15‐40), ALT 30U/L (9‐50)), while complete blood counts  and proband's younger brother (II‐3) by whole exome sequencing  in exon 3 of the g Figure . This miwww.hgmd.cf.ac.uk/) and MIM (https://www.omim.org). The heterozygous prevalence of this missense mutation is very low, 0.000025 in ExAC (http://exac.broadinstitute.org) and 0.000022 in gnomAD (http://gnomad.broadinstitute.org) respectively. There is no homozygous prevalence of this missense mutation reported in both databases.This mutation is co‐segregated well among the affected patients and unaffected carrier in this family but absent in unaffected family members and 200 healthy control. We have not identified this mutation in Human Gene Mutation database . Replacement of basic, polar and positive charged arginine by sulphur containing, non‐polar and neutral cysteine at the evolutionarily highly conserved position (p.Arg776) possibly exerts a recessive negative effect which finally results into muscular dystrophy. The positive charge of arginine forms favourable electrostatic interactions with the negative phospholipid head‐group. Hence, arginine mutated to cysteine will perturb the normal electrostatics. In addition, R776 is the first cytoplasmic residue of β‐dystroglycan. R776 also forms a part of the β‐dystroglycan nuclear localisation signal.The novel missense mutation in DAG1 mutation is associated with primary dystroglycanopathy in limb‐girdle muscular dystrophy and muscle‐eye‐brain disease.In addition, both genotypically and phenotypically, dystroglycanopathy is extremely heterogeneous. The clinical diagnosis of these three generation Chinese family with Limb‐girdle muscular dystrophy has been done based on the classification system In addition, due to extreme genotypic and phenotypic heterogeneity, clinical diagnosis through genetic screening for the patients with dystroglycanopathy, is really a big challenge. In this study, we performed whole exome sequencing (WES) for identifying the candidate gene with novel mutation. Our present study strongly emphasises the significance of WES for rapid, accurate and cost‐effective approach for identifying the causative gene with pathogenic mutation.DAG1 gene associated MDDGC9 is a very rare dystroglycanopathy and till now less than 10 cases reported worldwide. Here, the clinical symptoms of patients are also very different from those previous reports as the clinical phenotype of our studied patient is very mild and very slowly progressing with late age of onset. To be the best of our knowledge, this is the first description of a potentially pathogenic mutation of the DAG1 gene associated with MDDGC9 in China.However, The authors confirm that there are no conflicts of interest.Y.D., S.L. conducted acquisition, analysis and interpretation of data, and drafted and edited the manuscript. S.B., L.Cu., L.C., Y.D. and S.L. supervised all aspects of this study including study design, data interpretation, and drafted and edited the manuscript. X.D., Y.Z. and H.R. made WES pipeline and analysed the data. Y.D., Y.G. and H.Y. selected patients and performed WES. Y.D. and S.L. collected clinical information of the patient. S.B., L.Cu., L.C., C.L. and Y.D. supervised manuscript preparation and edited the manuscript.


LYRM4 gene codifies for ISD11, a small protein (11 kDa) acting as an iron‐sulfur cluster, that has been recently confirmed as a disease‐causing gene for mitochondrial disorders. We present a 4‐year‐old girl patient, born from non‐consanguineous healthy parents, with two episodes of cardiorespiratory arrest after respiratory viral illness with progressive decreased activity and lethargy, at the age of 2 and 3 years. She was asymptomatic between crisis with regular growth and normal development. During acute events of illness, she had hyperlactacidemia (maximum lactate 5.2 mmol/L) and urinary excretion of ketone bodies and 3‐methylglutaconic acid, which are normalized after recovery. A Next Generation Sequence approach with a broad gene panel designed for mitochondrial disorders revealed a novel probably pathogenic variant in homozygosity in the LYRM4 gene [p.Tyr31Cys (c.92A>G)] with Mendelian segregation. Functional studies in the skeletal muscle confirmed a combined deficiency of the mitochondrial respiratory chain . To our knowledge, this is the third case of LYRM4 deficiency worldwide and the first with 3‐methylglutaconic aciduria, not reported in any Fe‐S cluster deficiency. Remarkably, it appears to be no neurological involvement so far, only with life‐threating acute crisis triggered by expectably benign autolimited illnesses. Respiratory chain cofactors and chaperones are a new field of knowledge and can play a remarkable effect in system homeostasis.In the era of genomics, the number of genes linked to mitochondrial disease has been quickly growing, producing massive knowledge on mitochondrial biochemistry.  As an early intervenient in ISC biogenesis, ISD11 deficiency can have a negative interference in multiple systems and results in combined OXPHOS deficiency (MIM#615595—COXPD19). ISD11 also plays a key role in iron homeostasis as a controller of the intracellular iron trafficking, and its deficiency leads to mitochondrial iron overload.ISD11 is a 11 kDa protein found at both mitochondria and nucleusLYRM4 deficiency.To our knowledge, there are only two published patients with 2Chromatography of urinary organic acids was performed by gas chromatography mass spectrometry; after acidification, salting and extraction with trimethylsilyl in a sample volume are equivalent to a creatinine volume of 5 mmol/L. Two internal standards were used .Histological studies were performed on tissue from the deltoid muscle collected by open biopsy under general anesthesia. Sample was preserved in saline‐moistened gauze and immediately processed. Histological studies included modified Gomori trichrome, cytochrome oxidase (COX), and succinate dehydrogenase stains.Spectrophotometric measurements of respiratory chain enzymes and citrate synthase were carried out in skeletal muscle homogenates as previously described,http://www.1000genomes.org) and Exome Variant Server databases (http://evs.gs.washington.edu) were filtered out]. Sanger sequencing was used to validate the mutation and to study Mendelian segregation in parents.Genetic studies were performed by a Next Generation Sequence (NGS) approach with a gene panel designed for mitochondrial disorders including 209 nuclear genes known to be associated with mitochondrial diseases Table . Genes o3We present the case of a 4‐year‐old girl, first child of a non‐consanguineous healthy Portuguese couple, with normal growth and neurodevelopment. She had an episode of cardiorespiratory arrest at the age of 2 years after a respiratory viral infection. Microbiological studies identified respiratory syncytial virus (RSV) in respiratory secretions and Human herpesvirus 6 (HVV6) in cerebrospinal fluid (CSF). Work‐up during recovery showed a persistent increased lactate . Her electrocardiogram, echocardiogram, and brain magnetic resonance imaging (MRI) were normal. She was discharged with pending metabolic results and was lost in the follow‐up.At the age of 3 years, she was admitted to our hospital after cardiac arrest following acute respiratory insufficiency. Similar to the first episode history, parents reported nasal discharge, cough, fever, and reduced appetite for the previous 4 days, with a progressive decrease of activity. That day she had been lethargic and presented with difficulty to breath prior to the cardiorespiratory arrest.After resuscitation, apart from pulmonary auscultation with rhonchi, her physical examination was normal. Diagnostic investigation revealed persistently high lactate , normal blood and CSF counts (lactate in CSF was not performed), negative C‐reactive protein, and sterile blood culture. She had no hypoglycemia and her initially elevated liver enzymes and creatine kinase (CK)  rapidly returned to normal values. Chest X‐ray showed no pneumoniae and RSV was isolated in respiratory secretions. Cardiac evaluation  was normal and an immunodeficiency was excluded .Plasma amino acids were normal and acylcarnitines showed only low free carnitine  with normal acetylcarnitine. Urinary organic acids on admission showed ketonuria and moderate excretion of 3‐methylglutaconic acid (3‐MGA) with 154 μmol/mmol creatinine (NR <19 μmol/mmol creatinine) and mild excretion of 3‐hydroxyisovaleric acid (3‐OH‐IVA) with 85 μmol/mmol creatinine (NR 10.4‐67.0 μmol/mmol creatinine) and 3‐hydroxymethylglutaric acid (3‐HMG) with 93 μmol/mmol creatinine (NR 6.2‐49.7 μmol/mmol creatinine), as shown in Figure At this point, she enrolled in a research study for the diagnosis of mitochondrial disorders by an NGS approach. Retrieving information from the first episode showed that 2 weeks after the cardiorespiratory arrest she had mild excretion of 3‐MGA (27 μmol/mmol creatinine) accompanied by multiple other metabolites, such as 4‐hydroxyphenyllactic and 3‐hydroxyadipic acids and traces of 3‐methylglutaric, 2‐OH‐IVA, and sebacic acids.After 5 days on mechanical ventilation and vasoactive support, she was weaned to spontaneous breathing and after 24 hours on noninvasive ventilation. Asymptomatic and with a normal neurologic examination, she was discharged after 2 weeks under carnitine per os. After recovery, lactate and organic acids completely normalized as well as carnitine levels, even after suspension of carnitine supplementation.A close follow‐up was guaranteed. During the subsequent episode of fever with reduced oral intake, without evidence of significant general status or neurological deterioration, CK levels and liver enzymes were normal but her urinary profiling again revealed marked ketosis and mild excretion of lactate, 3‐MGA (129 μmol/mmol creatinine), 3‐OH‐IVA (162 μmol/mmol creatinine), and 3‐HMG (102 μmol/mmol creatinine) . Brain MRI evaluation was delayed pondering the general anesthetic risk vs her normal neurological status.4LYRM4 gene [p.Tyr31Cys (c.92A>G)], with heterozygous parents for the same variant. The pathogenicity was predicted bioinformatically by SIFT, PolyPhen‐2, and MutationTaster as probably pathogenic variants. The p.Tyr31Cys variant affects an amino acid that is highly conserved in different species through evolution  and confirmed the diagnosis with the evidence of a combined deficiency of complexes I, II, and IV with 40, 23, and 31% of residual activity, respectively  excretion of 3‐MGA during acute events, not previously reported in patients with any Fe‐S cluster deficiency. 3‐MGA‐uria can be found in various inborn metabolic disorders commonly along with excretion of other metabolites . It can be a feature of mitochondrial disorders present in about 10% of patients,TAZ gene) and MEGDEL syndrome (SERAC1 gene); (b) mitochondrial membrane‐associated disorder: Costeff syndrome (OPA3), TMEM70 defect (TMEM70), and DCMA syndrome (DNAJC19); (c) unknown mechanism: Sengers syndrome (AGK), CLPB deficiency (CLPB), TIMM50 deficiency (TIMM50), HTRA2 defect (HTRA2), and short‐chain enoyl‐CoA hydratase deficiency (ECHS1).Inborn errors of metabolism with 3‐MGA‐uria as discriminative feature  can be classified as primary or secondary 3‐MGA‐uria.This is the first case of Fe‐S cluster with 3‐MGA‐uria, extending the differential diagnosis of 3‐MGA‐uria as the presenting feature.Being a disorder related to iron metabolism, with the possibility to induce iron overload on tissues and storage depletion, evaluation of ferritin, transferrin, soluble transferrin receptor, or hepcidin may help understand the impact on the dynamics of iron metabolism, despite no evidence of impaired iron homeostasis in our patient so far.l‐cysteine or N‐acetylcysteine is supported by studies on patients with mitochondrial disorders with some benefit in those with translation‐related defects.Since cysteine acts the major sulfur donor, supplementation with this nonessential amino acid can be beneficial, especially during periods of higher energy requirement. The rationale for treatment with Raised awareness and a low threshold to hospital admittance in patients with ISD11 deficiency may be crucial to improve survival. Routine neurological and neurodevelopment evaluation must be granted.This case illustrates the importance of metabolic work‐up to be performed as soon as possible, while the patient is acutely ill, as it may be a unique opportunity to diagnose a critical condition. Moreover, life‐threatening events disproportional to symptoms and medical history should elicit the possibility of energy‐related inherited metabolic disorder. Acute presentations of mitochondrial disorders will become more common with the improved diagnostic yield of genetic studies.M.P.C. was involved in patient diagnosis, literature review, and drafting of the manuscript. J.C. was involved in patient follow‐up and investigation, and drafting of the manuscript. A.D. worked on chromatography of urinary organic acids, interpretation, and figure construction. C.N. worked on genetic studies and interpretation. A.B. and E.M. were involved in patient's work‐up and did critical manuscript review. L.V. worked on functional and genetic studies interpretation and did critical manuscript review.The authors declare that they have no conflict of interest.The study “Genetic Defects of Mitochondrial Diseases: a Next Generation Sequencing Approach” (FCT PTDC/DTP‐PIC/2020/2014) was approved by the Ethics Committee of Centro Hospitalar do Porto (REF. 2016.194(164‐DEFI/153‐CES). Informed consent from patient's parents was obtained.Table S1 Nuclear genes involved in mitochondrial disordersClick here for additional data file.


Temperature monitoring in the perioperative periods is important in order to avoid both hyperthermia and hypothermia. In our pilot study, we evaluated the usefulness of Bair Hugger™ temperature monitoring system (BHTMS), a forehead deep temperature monitoring system, in the neck and chest under general anesthesia.Eso) were monitored and analyzed.After approval from the Sapporo Medical University Research Ethics Board, 30 female patients scheduled for laparoscopic surgery were enrolled in this study. Patients were divided into three groups, depending on the attachment regions of BHTMS sensor. Temperatures obtained from the three regions and each esophageal temperature (TEso was + 0.05 °C above TEso (2SD ± 0.35 °C), and that between temperature obtained from the chest and TEso was − 0.55 °C above TEso (2SD ± 0.55 °C).A Bland-Altman plot showed that the mean bias between temperature obtained from the neck and TBy using the BHTMS sensor in the neck region, it is possible to monitor core body temperature seamlessly and with high reliability. These results may suggest that the use of BHTMS has high versatility in measuring perioperative core body temperature.UMIN000016802 Registered 15 March 2015).This study was approved by the Sapporo Medical University Research Ethics Board (2015: No. 262-149) and registered with UMIN Clinical Trial Registry ( Temperature monitoring in the perioperative periods is important in order to avoid both hyperthermia and hypothermia. Malignant hyperthermia, the phenomenon of hyperthermia during use of volatile anesthetics and succinylcholine, requires monitoring and urgent treatment for the drastic temperature elevation , 2. PeriDuring surgery, core body temperature is usually monitored in the rectum, bladder, and ear. However, temperatures obtained from these body regions sometimes poorly reflect the real values and drastic changes in core body temperature –12. As aHence, we attached the forehead sensor to other body regions and continuously compared with the esophageal temperature which is highly reliable for core body temperature , 17. TheN = 10), neck group (N = 10), and anterior chest group (N = 10) due to different attachment regions of BHTMS sensor. None of the patients received premedication before entering the operation room. In the operation room, a 20G intravenous catheter was inserted into the light forearm and bicarbonate Ringer’s solution fluids warming at 37.0 °C started to infuse. General anesthesia was induced by using propofol (1.5–2.0 mg/kg) and fentanyl (1.0–2.0 μg/kg) and the tracheal intubation was performed after injecting 0.9 mg/kg rocuronium. After intubation, esophageal temperature probe  was properly inserted into the esophagus under observing by McGrath®MAC . We located the temperature probe tip in the lower esophagus and the distance was predicted in advance by referring to patients’ chest radiograph. And then, BHTMS sensor was placed on the light forehead in forehead group, above the left common carotid artery in the neck group, and on the 4 left sternal borders in the anterior chest group. After setting the thermometers, we performed the ultrasound-guided transversus abdominis plane block or quadratus lumborum block using 0.375% ropivacaine (dose within 3.0 mg/kg) as postoperative pain control. These processes of general anesthesia induction were performed by either two experienced anesthesiologists. Anesthesia during the operation was maintained with 1.5% sevoflurane in 3 L/min air and 1 L/min oxygen with 0.1–0.2 μg/kg/min remifentanil continuous administration. The ambient temperature was maintained approximately at 23.0 °C and the humidity at 40% during operation. Approximately 60 min before the end of the surgery, 1000 mg acetaminophen was intravenously administrated. Esophageal temperature (TEso) and temperature obtained from three locations  were monitored and continuously recorded to a laptop computer at 5-min intervals until just before discontinuation of sevoflurane exposure. Anesthesiologist in attendance appropriately used a forced-air warming system  and blankets , warmed the patients at 43 °C during general anesthesia.This study was approved by the Sapporo Medical University Research Ethics Board , and written informed consent was obtained from each participant. The trial was registered at the UMIN Clinical Trials Registry (UMIN000016802) before patient recruitment. Eligible participants were 30 female patients between 21 and 80 years old who had scheduled laparoscopic surgery under general anesthesia except for upper gastrointestinal tract surgeries. The patients enrolled in this study  have no esophageal lesions and/or no abnormalities in the region of attachment and were not expected to be suffering from massive hemorrhage during surgery. We divided the participants into three groups—the forehead group  between TEso and THead, TEso and TNeck, and TEso and TChest. Considered to several past articles about body temperature monitoring, the mean value of the difference (bias) < 0.4 °C and 2 standard deviations (SD) < ± 1.0 °C was defined statistically significant in this study. All statistical analyses were performed using Prism software version 6 for Windows .One-way ANOVA was used to compare the patients’ background differences of age, height, weight, and body mass index (BMI). Kruskal-Wallis test was used to compare the differences of the length of operation time, length of anesthesia time, blood loss, and estimated water balance. Eso and THead, TEso and TNeck, and TEso and TChest, respectively. No monitoring defect caused by electronic devices was observed in this study. There were no significant differences in age, height, weight, BMI, length of operation, length of anesthesia, blood loss, and estimated water balance in each group (Table r) in each comparison. Pearson’s correlation coefficients were 0.81, 0.86, and 0.84 between TEso and THead, TEso and TNeck, and TEso and TChest, respectively, indicating a strong correlation in all comparisons  to − 0.06 °C (upper coefficient limit), it is necessary to consider how to interpret numerical values under real clinical use. One reason for systematic error with anterior chest application might be influenced by the effect of the rib bones and pericardial fatty tissue, and movements of the thoracic cage and aerated lung during breathing.Our study has certain limitations. First, we monitored and analyzed core body temperature within a limited range, almost between 35.5 and 38.0 °C, in laparoscopic surgery cases. Thus, we cannot judge whether our monitoring method is proper in hyperthermia cases over 38.0 °C and hypothermia cases below 35.5 °C. Second, we collected 10 cases and data from each group, but the sample size was too small. Third, we did not measure peripheral temperature in this study, to serve as a comparison. We need further verifications to resolve these limitations; however, the knowledge obtained from this study would be one option in monitoring core body temperature.To conclude, it is possible to monitor core body temperature seamlessly and with high reliability by using the BHTMS in the neck region. Usefulness of BHTMS in the neck is equivalent to that in the original frontal head region. The value measured in the chest has a systematic error, and it is necessary to judge by considering the LOA. In addition, these results from this pilot study suggest that using BHTMS may be more versatile in measuring perioperative core body temperature.


Maternal family history of diabetes was significantly and positively associated with birth weight in grandchildren, we aim to assess the effect of grandparental diabetes on the grandchild’ body mass index (BMI) at infancy peak (IP) and obesity status at age 2.In our study, family diabetes mellitus (DM) information from Ma’anshan Birth Cohort Study (MABC) were gathered. For children, height and weight were retrieved from medical records. BMI at 6 observations  was plotted for every child. Onset of IP was determined by visual inspection. BMI at age 2 was categorized according to WHO Child Growth Standards as normal, overweight or obesity. The association between maternal grandfather’ diabetes and the grandchild’ BMI at IP and BMI at age 2 were tested using linear regression models and logistic regression models, respectively.2, and 6.6% of the children were obesity at age 2. Maternal grandfather with DM could significantly increase the IP BMI values , and was associated with obesity status at age 2 , but maternal grandmother and paternal grandparents were unrelated.In our sample, about 6% of the maternal grandfather had DM, mean of infancy BMI peak was 18.37 kg/mThese results suggest that DM in maternal grandfather may be a risk factor for the grandchild high BMI at peak and obesity at age 2. Unfavorable body mass index (BMI) at the early stage of human development can induce permanent health risks. The prevalence of childhood obesity has brought a heavy cost burden, result in later complications including diabetes, metabolic syndrome, and cardiovascular diseases –3. One oSeveral studies have paid close attention to the intergenerational cycle of diabetes mellitus and obesity . After aThere was evidence suggesting that maternal family history of diabetes were significantly and positively associated with gestational diabetes mellitus (GDM) , and GDMn = 162), plural gestations (n = 39), without children gender information (n = 6), without grandparents exposure information (n = 137), unbalanced BMI data at 0–1.5 years of age (n = 769) or missed children’s BMI data at age 2 (n = 504) were excluded from this study  initially focused on the relationship between maternal factors during pregnancy and the development of the child. We looked into more information about lifestyle characteristic of father and grandparents. In this study, 3474 gravidas consecutively enrolled from May 2013 to September 2014 in Ma’anshan Maternal and Child Health(MCH) clinics when they did first physical examination during pregnancy. Details on the design and recruitment procedures have been described elsewhere . Among tThe information of ancestors and children was gathered by “Maternal and child health questionnaires”, which covered family medical history, sociodemographic characteristics, childhood lifestyles etc. DM in grandparents was based on self-reported data from questionnaires in which the mother was asked whether any of her biological parents and/or her parents in law had diabetes, and response categories for diabetes history were ‘yes’, ‘no’ and ‘unclear’. Children aged 0 months, 3 months, 6 months, 9 months, 12 months, 18 months and 24 months were measured by physician, therefore, their height and weight was extracted from medical records. BMI was calculated via dividing weight in kilograms by the square of height in meters.Individual -specific BMI peaks were determined by visual inspection. It has been proven that visual inspection appears to be preferable than polynomial models when estimating the milestone of BMI trajectories . Onset on = 2361or 2626) without adjusting for feeding pattern, some maternal health parameters or some socio-economic covariates gave similar results, all these variables were adjusted for more accurate and more reliable.Based on other evidence, the following variables that might affect the physical development of children were considered: infant gender (boy or girl), feeding pattern, monthly income(U.S. dollar), maternal education, gestational diabetes mellitus , hypertension disorders complicating pregnancy (HDCP) , maternal age at birth, maternal pre-pregnancy BMI(continuous variable), paternal pre-pregnancy BMI(continuous variable), gestational weight gain(continuous variable), production times(first or more than once), delivery mode, birth term (pre-term or born in time), birth weight. Although analysing the data  were applied to estimate the association between variables and BMI values at IP; We examined differences in subject characteristics amongst BMI classes by using χ2 tests. We used linear regression models to access the association between ancestors’ DM with grandchildren’ BMI at IP, and the association between maternal grandparents with DM and the grandchild’ BMI classes at age 2 were tested using multinominal logistic regression models. The associations were defined significant if P < 0.1. All analyses were conducted in SPSS 23.0.Descriptive statistics were used to examine the relationship between ancestral factors and early childhood BMI status. 2(SD = 1.56) and occurred at 7.86 months. And about 6.6% of the children were obesity at age 2. The variables and BMI at IP or obesity status at age 2 of the 2361or 2626 grandparents-grandchild pairs are presented in Table P = 0.006, OR = 1.80, 95% CI: 1.18 ~ 2.74). The mean values of BMI at IP by maternal grandfather category were 18.36 kg/m2 and 18.66 kg/m2; and the percentages of children with obesity by maternal grandfather category were 4.3 and 8.7% for maternal grandfathers with normal status and diabetes mellitus, respectively. The difference between both groups was statistically significant .Age and BMI at IP were not identifiable in 4.9% of our sample, mean of infancy BMI peak was 18.37 kg/mβ = 0.31, 95% CI = 0.03~0.58), a similar trend when the estimating was performed using a multivariate analysis ; the results suggest that BMI increases as when maternal grandmother and paternal grandparents had diabetes mellitus, however, the analysis did not reach a statistically significant difference.The association of grandparents’ diabetes mellitus and children’ BMI at IP are shown in Table OR = 1.92, 95% CI = 1.08~3.39), no statistical difference between maternal grandfather’ diabetes mellitus and children overweight ; however, maternal grandmother and paternal grandparents with diabetes mellitus was unrelated to children BMI status at age 2.Children’ obesity and overweight were compared with normal weight across different ancestor exposure  demonstrated that family history of diabetes was associated with the prevalence of adults obesity as defined with the use of BMI [n = 12,076) revealed that the grandchildren of maternal grandparents with DM were more likely to have higher birth weight than those grandchildren of non-grandparental diabetics [Previous studies have only examined associations of GDM or other maternal factors with offspring BMI status and infancy BMI peak, but not grandparental DM. In concordance with our study, a cross sectional national survey in Sri Lanka  the grandparents were affected with. What’s worse, it is regrettable that our study has no concern with the population with undiagnosed diabetes or prediabetes. After all, a significant proportion remaining undiagnosed (over 60% were unaware of their diagnosis) in China , partly In conclusion, we found that DM in maternal grandfather may be a risk factor for the grandchild high BMI at infancy peak and obesity at age 2, and these population epidemiological findings may to some extent draw attention to trans-generational effects of diabetes and obesity.


With unique topological structure, GYs display unusual semiconducting properties, especially in the aspects of charge mobility and electron transport. Among the members of the GY family, only graphdiyne (GD) can be successfully synthesized in large quantities. The advanced properties of GD make it promising in various applications. Here, the recent progress in the synthesis of GD and GD‐based composites is reviewed as well as their applications in photorelated and electrocatalytic applications. It is hoped that this Review will promote the development and applications of carbon chemistry.As a rising star of carbon allotropes, graphynes (GYs) merely consist of sp‐ and sp Due to the larger diameters of carbon rings in the structure of GD, it is expected to exhibit a better charge and mass mobility as compared with graphene.5 cm2 V−1 s−1, comparable with that of graphene (≈3 × 105 cm2 V−1 s−1), indicating its excellent electron transport property.When the spAs known, the traditional nanomaterials, such as semiconductors, have their limitation in energy‐related applications due to their poor capability of charge separation and low transition efficiency.Herein, we summarized the methods in preparing 2D GD and GD‐based composites as well as their recent progress in photorelated and electrocatalytic applications. The challenges and prospect of GD are also discussed based on our understanding.2In respect to the excellent properties of GD, extensive efforts have been devoted to exploring their synthesis and composition with various functional materials for promising energy‐related applications. We here mainly elaborate their synthetic methods and corresponding properties.2.1Figure5 and 5.41 × 105 cm2 V−1 s−1, respectively, which are even larger than that of graphene (≈3 × 105 cm2 V−1 s−1).Graphynes (GYs) were first put forward by Baughman et al. early in 1987, but no detailed investigation has been reported.GD, a member in GY family that consists of diacetylene groups and benzene ring, was first proposed by Haley,2.1.1Although some frameworks were obtained at early time, the self‐standing GD sheets were not successfully synthesized until 2010.Synthesis of 2D GD: The large‐scale synthesis of 2D GD was achieved via crosscoupling reaction of hexaethynylbenzene (HEB) on the copper foil surface in pyridine through solution polymerization  films were prepared by a liquid/liquid interfacial synthesis method.Synthesis of 0D, 1D, and 3D GD: 0D GD nanoparticles were obtained by a simple grind‐sonication method.1D GD nanowires were prepared on well‐defined silver surfaces by surface‐confined coupling of specifically designed precursors on the basis of covalent coupling and supramolecular concepts.3D GD sponge was synthesized from the skeletons of melamine sponge (MS) via a facile in situ Glaser–Hay coupling.In addition to sponge, abundant and inexpensive diatomite was also employed as template by Li et al. to construct freestanding 3D GD.Later, Shang et al. constructed a 3D all‐carbon mechanical and conductive GD networks by a novel in situ weaving strategy on Si anode at ultralow temperature (25 °C).However, since most GD structures are inevitably grown on copper substrates, it is quite complicated to transfer them onto some specific substrates, which limits the application of GD on functional devices. To solve this issue, Zhang and co‐workers reported a feasible route to achieve in situ growth of GD nanowalls on various substrates by enclosing the substrates into a copper foil.2.1.2Few‐layer GD with larger exposed surface is supposed to possess better electron transfer properties compared with its bulk counterpart.FigureAs for lithium ion intercalation method, GD was used as cathode in a lithium cell and lithium foil was used as the anode. By optimizing the cutoff voltage to induce the intercalation of Li ions between GD layers , well‐dispersed ultrathin GD sheets were obtained after a postsonication treatment Figure3a,b.542/Si substrate. After immersing the substrate into precursor solution, ultrathin GD film was obtained via controlled polymerized growth in the presence of CuCl catalyst. The thickness is varied from 3 to 20 nm by controlling the concentration of monomer. Li et al. also prepared an ultrathin β‐GD‐like film by Eglinton coupling reaction through template synthesis due to the strong π–π interaction between β‐GD and graphene.With respect to template synthesis, graphene can serve as a template to grow ultrathin GD films.Previous methods cannot avoid the random encounter of HEB precursor and the catalysts, leading to the inevitable formation of GD nanoparticles besides nanosheets. To overcome this disadvantage, interfacial polymerization method has been developed.2.1.3Recently, numerous efforts have been devoted to the design, fabrication, and modification of various GD‐based functional composites for energy conversion and environmental protection. The hybrid materials can synergistically take advantage of each component to further boost the overall catalytic activity. In the following parts we will introduce the synthesis methods in detail with classification based on the dimensions of applied materials, such as element‐hybrid (doping) and material‐hybrid . The synthetic methods mainly involve simple solution mixing, hydrothermal reaction, oxidation‐reduction, impregnation, ion‐beam sputtering, and the thermal pyrolysis method.The meaning of functionalization of these 2D carbon materials is significant. For photochemistry in semiconductors, when the electron in the valance band (VB) is excited into conduction band (CB), a hole is generated in the VB. However, the generated holes and electrons in excited states are not stable and they are easy to recombine. Thus, designing photocatalysts to suppress the recombination of holes and electrons and improve photocatalysis performance is highly desired. One common strategy is to produce hybrid photocatalyst by loading semiconductors on various carbon substrates. Due to the stronger electron affinity and transfer capability, the recently emerged 2D carbon allotropy, namely GD, is considered as a better assistant in photocatalysis compared with other carbon substrates. Besides, most surfaces of carbon materials are chemically inert, which need further modification to optimize the electronic properties for electrochemistry. Doping with other light elements can effectively break the electron neutrality and promote the intrinsic catalytic activity. In this section, we summarize the recent development in the preparation of various GD‐based composites for photo‐ and electrochemical conversion.0D/GD Hybrid: Extensive efforts have been devoted to developing highly efficient and stable catalysts by compositing various 0D metals or semiconductors with GD, such as Ag/AgBr/graphene oxide (GO)/GD, P25‐GD, TiO2‐GD, Pd/GD, PtNP‐graphdiyne nanosheet (GDNS), Fe/GD, and Pt‐GD. For example, Zhang et al. prepared Ag/AgBr/GO/GD by mixing the chloroform solution of Ag precursor (AgNO3) with GO/GD suspension by an ultrasonic homogenizer.Figure2 nanoparticles by forming a strong chemical bond of Ti‐C. Based on that, Yang et al. synthesized TiO2 (001)‐GD by a simple hydrothermal method with the perfect interaction between the crystalline TiO2 nanosheets and GD.Besides as the template, GD can serve as both reductant and stabilizer for direct synthesis of ultrafine Pd nanoparticles (NPs).Moreover, some physical techniques have also been adopted to fabricate the 0D/2D hybrid. For example, Ren et al. has reported the ion‐beam sputtering approach to deposit Pt nanoparticles onto pre‐exfoliated GD substrates.1D/GD Hybrid: The fabrication of 1D/GD hybrid materials is rarely reported. One successful example is vertically growing NiCo2S4 nanowires on the surface of 3D GD scaffolds , other carbon materials and even metal composites, to achieve better catalytic performance.2 nanosheets . Through the similar solvothermal method, Han et al. prepared a metal‐free 2D/2D heterojunction composed of GD and g‐C3N4.Yu et al. reported that the nitrogen‐doped GD (NGD) with porous structure and excellent conductive network can be used as the robust support for the growth of MoSs Figure e,f.67 ByHui et al. introduced porous GD nanolayers on the surface of iron carbonate hydroxide (FeCH).Doping: Besides the hybridization with different materials, doping is another effective method to enhance the catalytic activity of GD. N atom is one of the commonly reported doping atoms. The N atoms could be doped into the carbon structures, existing as three common bonding forms: pyridinic N, pyrrolic N, and graphitic/quaternary N.3, and pyridine. For example, nitrogen‐containing carbon nanolayers on GD nanostructure were synthesized by one‐step pyrolysis of iron salts and polyaniline loaded onto GD nanocomposite.3 atmosphere can result in the formation of N′N‐GD.The most efficient and well‐studied NGD catalysts can be obtained by reacting GD with various N‐containing precursors at high temperature, such as aniline, NHThe bottom‐up synthetic strategy can also be used to obtain GD doped with other heteroatoms. For example, Wang et al. synthesized boron‐GD (BGD) and chlorine‐substituted GD (Cl‐GD) in the presence of 1,2,3‐trichlorineborane and 1,3,5‐trichlorinebenzene by Glaser–Hay coupling reaction.2.22.2.1I–V curves in different current transport directions . GY, GY‐3, and GY‐5 show similar transport properties  are used to study the electron transport properties of GYs. GYs display a semi‐conductive characteristic along zigzag direction and metallic characteristic along armchair direction, which differs a lot from graphene.ns Figure5a–c. T5 cm2 V−1 s−1) at room temperature, while its hole mobility is an order of magnitude lower.For bulk GD, its electronic properties are also investigated using DFT, which are associated with its stacking configurations.4 cm2 V−1 s−1), which is also greatly larger than that of the hole mobility. It is found that the charge mobility increases with the width of GDNRs, and the divan‐edged GDNRs have larger mobility than the zigzag edged GDNRs. In addition, both the zigzag and armchair GDNRs display semi‐conductive characteristic.As for GD nanoribbons (GDNRs), the electron mobility at room temperature is about 10Bilayer GD (AB style) possesses a direct bandgap of 0.35 eV, whereas trilayer GD (ABA style) has a bandgap of 0.18–0.33 eV. Moreover, the bandgaps of the semiconducting bilayer and trilayer GD generally decrease with increasing external vertical electric field, irrespective of the stacking style.2.2.2x‐direction, the in‐plane stiffness and Young's modulus increase and finally approach constant values.n.−1, which is equivalent to a Young's modulus of 375 GPa if a thickness of 0.320 nm is assumed.The elastic properties (in‐plane stiffness and Young's modulus) of GD are studied using a self‐consistent charge density functional tight‐binding method. The results indicate that as the length of the introduced acetylene links in GD increases in the Mechanical properties of GYs with different number of acetylenic linkages are also investigated by molecular dynamics simulations. Yang et al. found that in the armchair loading case, the fracture strain remains nearly unchanged whereas the ultimate strength degrades gradually with longer acetylenic chains.2.2.3Luo et al. investigated the optical properties of several possible GD bulk structures.First principle calculations with generalized gradient approximation were used to analyze the optical properties of armchair and zigzag GY nanotubes (GNTs) doped with boron (B) and nitrogen (N) atoms.3The efficient interfaces constructed between GD and semiconductors can result in enhanced charge transfer and improved performance. In this case, GD and its composites have been widely utilized in photocatalysis, solar cells, and electrocatalytic reactions.3.1Semiconductor photocatalysts have drawn much attention in environmental remediation and their charge generation, separation and transport efficiency determine the overall performance.In this part, three main applications of GD‐based composites are reviewed, including photocatalytic degradation, photocatalytic reduction, and photocatalytic hydrogen generation.3.1.12, are easy to recombine. GD, as a novel form of carbon materials, possesses many exceptional properties, including high surface area and high electron mobility. GD‐based semiconductor photocatalysts can embrace synergistic effects between the metal oxides and carbon materials, which widen light absorption range and suppress electron–hole recombination.In recent years, photocatalytic degradation plays an increasingly important role in dealing with environmental problems, such as water purification, air cleanup, and waste remediation. Photogenerated electrons and holes of separate semiconductors, especially TiO2 has attracted much attention in photocatalytic degradation of contaminants due to strong UV light absorption and photostability. Wang et al. reported a hybrid of commercialized TiO2 (P25) and GD for photocatalytic degradation of organic dyes ‐GD possessed the highest charge separation capability, the strongest oxidation ability, and the most abundant impurity levels ‐GD also remains longer than that of TiO2 (001)‐GR. The VB position of TiO2 (001)‐GD is lower than that of TiO2, indicating its higher oxidation ability. In addition, TiO2 (001)‐GD possesses positive and negative charges, and GD can introduce impurity levels to TiO2 more easily than graphene, resulting in abundant impurity levels. The photocatalytic degradation of MB is shown in Figure 2 (001), the first‐order rate constant k is estimated to be 0.0152 min−1 by normalizing temporal concentration change (C/C0) of MB with respect to time, whereas TiO2 (001)‐GR composite with 0.6 wt% GR shows a 1.28 times improvement (0.0195 min−1). Interestingly, for TiO2 (001)‐GD composite with 0.4 wt% GD, k is up to 0.0247 min−1, which is 1.62 times that of pure TiO2 (001) and the highest among all these TiO2 (001)‐based catalysts.To further understand the charge transfer mechanism, it is necessary to explore the interactions between the crystal facets of TiOs Figure c.61 In d2 by two orders of magnitude. Thangavel et al. synthesized the hybrid of ZnO‐GD via a hydrothermal method.To further increase the photocatalytic activity, ZnO nanoparticles were introduced because their electron mobility is higher than that of TiO3.1.2Since GD has lower reduction potential, it can be used as a reactant to reduce metal ions. Qi et al. reported that the GD has strong interaction with Pd, which can dramatically avoid the aggregation of atom, and serve as a substrate to deposit ultrafine Pd clusters.In addition, N doping can make the catalyst hydrophilic, and the resulting NGD could be employed as metal‐free photocatalyst.3.1.3Hydrogen energy, as a powerful and clean fuel source, has attracted great attention. Photoelectrochemical hydrogen production by semiconductor is regarded as a promising means in future.Figure−1), which is 2.6 times of pure CdS (1.6 mmol g−1). In addition, the photocatalytic activity of GD2.5 is also much higher than that of physical mixture of GD and CdS, indicating the great impact of forming CdS/GD heterojunction on improvement of the photocatalytic activity of CdS.Metal chalcogenides, especially CdS nanoparticles, have obtained much attention due to relatively negative CB position.Among the various structures of semiconductor catalysts, quantum dots (QDs) have been used as an alternative due to exceptional light‐harvesting ability and relatively large surface area. Li et al. prepared CdSe QDs/GD photocathode for the first time, where GD performs as the hole transfer layer Figure b.92 The 3.2Due to the unique electronic property, GD has been widely used in solar cells. The insertion of GD into solar cells can facilitate charge extraction and transport. Generally, GD can play three kinds of roles in solar cells, including counter electrode, electron transport layer, and hole‐transporting layer.3.2.13−/I−) redox couple of the electrolyte.As an alternate to silicon‐based solar cells, dye‐sensitized solar cells (DSSCs) have been extensively studied due to its low cost, high efficiency and easy operation, in which the counter electrode is the key part. The requirement for counter electrode in DSSC is that it should possess a higher catalytic activity in catalyzing the triiodide/iodide  increased from 13.5% to 14.8%  and ZnO films of perovskite solar cells  layer is critical for improvement of perovskite solar cells. Doping GD into HTM can greatly hinder the recombination of generated electrons and holes and improve charge extraction and transport. With this method, Xiao et al. demonstrated the hybrid of poly (3‐hexylthiophene) (P3HT) and GD can increase the cell performance owing to the formation of strong π–π stacking interaction.44.1With great efforts being devoted into studying non‐noble metal ORR catalysts, a novel class of metal‐free ORR catalysts has recently attracted researcher's attention.FigureZhang et al. prepared NGD using high‐purity ammonia.E1/2 of 0.87 V and Jk of 38.0 mA cm−2 at 0.75 V, even surpassing commercial Pt/C under alkaline conditions, and it showed comparable performance in acidic media  showed a high performance for HER.FigureIn order to further increase the atom utilization efficiency, single‐atom catalysts are explored. The isolated nickel/iron atoms were anchored on GD successfully.2 also showed a high activity and extraordinary stability (Figure 2 catalyst, showing an onset potential of ≈140 mV and Tafel slope of ≈54 mV dec−1 in HER.Besides the noble metals, the hybrid of GD nanosheets and MoSy Figure c,d.67 Ya5To summarize, this review provides an overview of the recent achievement in the synthesis, properties and up‐to‐date applications of GD and GD‐based composites in photorelated and electrocatalytic applications. This new carbon allotrope, i.e., GD, has excellent intrinsic electrical, mechanical, and optical properties. When combined with metal nanoparticles, semiconductors or doped with other elements, it can greatly enhance the charge separation and transportation during the photorelated or electrocatalytic conversion processes, bridging the gaps between semiconductors or metal nanoparticles by providing good electrical contact and low interfacial resistance. Their excellent performance in photo‐ or electrochemical conversion areas suggests they are a cost‐effective and superior alternative to the prevalent carbon materials, like graphene and carbon nanotubes.Though some initial successes have been achieved by GD‐based materials in photoelectric conversion and catalysis areas, this field is still in preliminary stage with significant challenges needed to be solved urgently. First, the few‐layer GD or other kinds of GYs possess more advantages than bulk materials, and the large scale fabrication of high‐quality and defect‐free ultrathin GD or other kinds of GYs still remains a challenge. Second, most studies on GD are still in the theoretical stages and some new members in the GY family with different percentage of acetylenic linkages and adjustable mechanical and electronic structures are anticipated to be obtained, such as 6,6,12‐graphyne. Third, the chemical functionalization of GD and other kinds of GYs, doping them with other elements, and hybridization them with other functional materials are still needed to be improved to endow them with desired bandgap structure and fascinating charge mobility, which play a vital role in a variety of electronic and optoelectronic areas. Fourth, to further develop both fundamental study and application process, the in‐depth understanding of the catalytic nature of GD and GD‐based composites is needed, which may be studied by theoretical simulations and in operando techniques, such as X‐ray adsorption spectroscopy or Fourier‐transform infrared spectroscopy. In a word, more efforts are needed to fully exploit the most valuable nature of these new 2D carbon allotropes. We believe that all the challenges and limitations will be overcome, and the bright future is near in the corner.The authors declare no conflict of interest.