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Philippe S Hwang,1 My-Linh Ma,1–3 Nora Spiegelberg,1 Catherine E Ferland1–5 1Shriners Hospital for Children – Canada, 2Research Institute – McGill University Health Centre, 3Department of Experimental Surgery, 4Department of Anesthesia, McGill University, 5The Alan Edwards Centre for Research on Pain, Montreal, QC, Canada Abstract: Conditioned pain modulation (CPM) paradigms have been used in various studies with healthy and non-healthy adult populations in an attempt to elucidate the mechanisms of pain processing. However, only a few studies so far have applied CPM in pediatric populations. Studies finding associations with chronic pain conditions suggest that deficiencies in underlying descending pain pathways may play an important role in the development and persistence of pain early in life. Twelve studies were identified using a PubMed search which examine solely pediatric populations, and these are reviewed with regard to demographics studied, methodological approaches, and conclusions reached. This review aimed to provide both clinicians and researchers with a brief overview of the current state of research regarding the use of CPM in children and adolescents, both healthy and clinical patients. The implications of CPM in experimental and clinical settings and its potential to aid in refining considerations to individualize treatment of pediatric pain syndromes will be discussed. Keywords: conditioned pain modulation, descending endogenous pain inhibition, chronic pain, pediatrics

Objectives: The aim of the study was to examine the association between Black, White, and Hispanic or Latino race and ethnicity and referral rejection from private postacute care facilities among hospitalized individuals with opioid use disorder (OUD)., Methods: In this retrospective cohort study, we linked electronic postacute care referrals from Boston Medical Center in 2018 to electronic medical record data, which we used to ascertain OUD status and race and ethnicity. Using multivariable logistic regression, we examined the association between Black, White, and Hispanic or Latino race and ethnicity and referral rejection, adjusting for individual-level characteristics including medication for opioid use disorder treatment type and for facility-level factors using facility random effects., Results: We identified 159 hospitalizations from 141 individuals with OUD referred to private postacute medical care, corresponding to 1272 referrals to 244 facilities. Hospitalizations comprised 53 (33%) non-Hispanic Black, 28 (18%) Hispanic or Latino, and 78 (49%) non-Hispanic White individuals. In adjusted analyses, referrals for non-Hispanic Black individuals had significantly higher odds of rejection compared to referrals for non-Hispanic White individuals (adjusted odds ratio 1.83, 95% confidence interval [1.24, 2.69], P = 0.002). There were no significant differences between referrals for Hispanic or Latino individuals and non-Hispanic White individuals (adjusted odds ratio 1.11, 95% confidence interval [0.67, 1.84], P = 0.69)., Conclusions: Among people with OUD referred to private postacute care in Massachusetts, non-Hispanic Black individuals were more likely to be rejected compared to non-Hispanic White individuals, demonstrating racism in postacute care admissions. Efforts to address discrimination against people with OUD in postacute care admissions must also address racial equity., (Copyright © 2024 American Society of Addiction Medicine.)

Background: Cell-free DNA (cfDNA) analysis offers an attractive noninvasive means of detecting and monitoring diseases. cfDNA cleavage patterns within a short range (e.g., 11 nucleotides) have been reported to correlate with cytosine-phosphate-guanine (CpG) methylation, allowing fragmentomics-based methylation analysis (FRAGMA). Here, we adopted FRAGMA to the extended region harboring multiple nucleosomes, termed FRAGMAXR., Methods: We profiled cfDNA nucleosomal patterns over the genomic regions from -800 to 800 bp surrounding differentially methylated CpG sites, harboring approximately 8 nucleosomes, referred to as CpG-associated cfDNA nucleosomal patterns. Such nucleosomal patterns were analyzed by FRAGMAXR in cancer patients and pregnant women., Results: We identified distinct cfDNA nucleosomal patterns around differentially methylated CpG sites. Compared with subjects without cancer, patients with hepatocellular carcinoma (HCC) showed reduced amplitude of nucleosomal patterns, with a gradual decrease over tumor stages. Nucleosomal patterns associated with differentially methylated CpG sites could be used to train a machine learning model, resulting in the detection of HCC patients with an area under the receiver operating characteristic curve of 0.93. We further demonstrated the feasibility of multicancer detection using a dataset comprising lung, breast, and ovarian cancers. The tissue-of-origin analysis of plasma cfDNA from pregnant women and cancer patients revealed that the placental DNA and tumoral DNA contributions deduced by FRAGMAXR correlated well with values measured using genetic variants (Pearson r: 0.85 and 0.94, respectively)., Conclusions: CpG-associated cfDNA nucleosomal patterns of cfDNA molecules are influenced by DNA methylation and might be useful for biomarker developments for cancer liquid biopsy and noninvasive prenatal testing., (© Association for Diagnostics & Laboratory Medicine 2024.)

The analysis of tissues of origin of cell-free DNA (cfDNA) is of research and diagnostic interest. Many studies focused on bisulfite treatment or immunoprecipitation protocols to assess the tissues of origin of cfDNA. DNA loss often occurs during such processes. Fragmentomics of cfDNA molecules has uncovered a wealth of information related to tissues of origin of cfDNA. There is still much room for the development of tools for assessing contributions from various tissues into plasma using fragmentomic features. Hence, we developed an approach to analyze the relative contributions of DNA from different tissues into plasma, by identifying characteristic fragmentation patterns associated with selected histone modifications. We named this technique as FRAGmentomics-based Histone modification Analysis (FRAGHA). Deduced placenta-specific histone H3 lysine 27 acetylation (H3K27ac)-associated signal correlated well with the fetal DNA fraction in maternal plasma (Pearson's r = 0.96). The deduced liver-specific H3K27ac-associated signal correlated with the donor-derived DNA fraction in liver transplantation recipients (Pearson's r = 0.92) and was significantly increased in patients with hepatocellular carcinoma (HCC) ( P < 0.01, Wilcoxon rank-sum test). Significant elevations of erythroblasts-specific and colon-specific H3K27ac-associated signals were observed in patients with β-thalassemia major and colorectal cancer, respectively. Furthermore, using the fragmentation patterns from tissue-specific H3K27ac regions, a machine learning algorithm was developed to enhance HCC detection, with an area under the curve (AUC) of up to 0.97. Finally, genomic regions with H3K27ac or histone H3 lysine 4 trimethylation (H3K4me3) were found to exhibit different fragmentomic patterns of cfDNA. This study has shed light on the relationship between cfDNA fragmentomics and histone modifications, thus expanding the armamentarium of liquid biopsy., Competing Interests: Competing interests statement:J.B., P.J., L.J., M.Y., K.C.A.C., and Y.M.D.L. filed patent applications based on the data in this study. Reviewer S.B. is an inventor on patents related to cfDNA mutation and methylation analysis technologies that have been licensed to Roche and Adela, respectively, and is a co-founder and has ownership in Adela.

Objective: To investigate the anatomical structure changes of the anterior segment and dynamic pupil changes in eyes with suspected primary angle-closure (PACS) under light and dark conditions, and their correlation with the occurrence of acute primary angle-closure (APAC). Methods: This cross-sectional study collected data from 37 PACS patients (66 eyes) who visited the ophthalmology clinic of the Second Affiliated Hospital of Harbin Medical University between September 2019 and March 2021. The patients included 12 males and 25 females, with an average age of (61.27±7.35) years. Of the 66 eyes, 58 had no history of APAC in the contralateral eye, while 8 had a history of APAC in the contralateral eye. Patients without a history of APAC in both eyes underwent the dark room prone provocative test (DRPPT), and the eyes were divided into three groups: DRPPT- (44 eyes), DRPPT+ (14 eyes), and APAC (8 eyes). The DRPPT+ and APAC groups were combined into the APAC+ group. All patients underwent ultrasound biomicroscopy (UBM) to measure the changes in various parameters under light and dark conditions, including anterior chamber angle 500 (ACA 500Δ ) and 1000 (ACA 1000Δ ), angle opening distance 500 (AOD 500Δ ) and 1000 (AOD 1000Δ ), and iris thickness 500 (IT 500Δ ) and 1000 (IT 1000Δ ). Anterior segment analysis was performed to capture the pupil changes under light and dark conditions, recording pupil diameter, maximum dilation speed (V max ), maximum constriction speed (V min ), and average speed (V m ). Results: There was no significant difference in the parameters between DRPPT+ group and APAC group ( P> 0.05). In the difference analysis, it was found that the V m value of DRPPT- group [(0.17±0.07) mm/s] was higher than that of APAC+ group [(0.13±0.06)mm/s], and the difference was statistically significant ( P <0.05). There was no significant difference in other parameters ( P >0.05). V max was positively correlated with temporal ACA 1000Δ and temporal AOD 1000Δ in all patients with PACS ( r =0.302, 0.260; P <0.05), V min was negatively correlated with temporal ACA 1000Δ and temporal AOD 1000Δ ( r =-0.338, -0.330; P <0.05). Conclusions: The dynamic changes in the anterior segment and pupil under different lighting conditions provide insights into the risk factors and potential predictive indicators for the occurrence of APAC in PACS patients.

In the human body, carboxylesterases (CEs) play crucial roles in xenobiotic metabolism and lipid homeostasis. But abnormal expression of CEs is highly associated with some diseases, such as hyperlipidemia, diabetes, and liver cancer. Therefore, it is of great importance to develop an efficient tool for the accurate detection of CEs in living organisms. Herein, an innovative near-infrared (NIR) fluorescent probe, TTAP - AB, was designed for CE detection based on the aggregation-induced emission (AIE) mechanism. This probe exhibits rapid response (2 min), excellent sensitivity (limit of detection = 8.14 × 10 -6 U/mL), and high selectivity to CEs. Additionally, owing to its good biocompatibility, the TTAP - AB probe enables the monitoring of dynamic changes in CE levels under drug-induced modulation in living cells and zebrafish. More importantly, the TTAP - AB probe was successfully employed to image liver tumors and assist in tumor resection through the real-time monitoring of CEs, indicating that TTAP - AB is promising to guide liver cancer surgery. Therefore, the TTAP - AB probe can not only enrich the strategies for CE detection in biological systems but also has great potential for some clinical imaging applications, including medical diagnosis, preclinical research, and imaging-guided surgery.

Background: Circulating tumor DNA (ctDNA) analysis has been applied in cancer diagnostics including lung cancer. Specifically for the early detection purpose, various modalities of ctDNA analysis have demonstrated their potentials. Such analyses have showed diverse performance across different studies., Methods: We performed a systematic review of original studies published before 1 January 2023. Studies that evaluated ctDNA alone and in combination with other biomarkers for early detection of lung cancer were included., Results: The systematic review analysis included 56 original studies that were aimed for early detection of lung cancer. There were 39 studies for lung cancer only and 17 for pan-cancer early detection. Cancer and control cases included were heterogenous across studies. Different molecular features of ctDNA have been evaluated, including 7 studies on cell-free DNA concentration, 17 on mutation, 29 on methylation, 5 on hydroxymethylation and 8 on fragmentation patterns. Among these 56 studies, 17 have utilised different combinations of the above-mentioned ctDNA features and/or circulation protein markers. For all the modalities, lower sensitivities were reported for the detection of early-stage cancer., Conclusions: The systematic review suggested the clinical utility of ctDNA analysis for early detection of lung cancer, alone or in combination with other biomarkers. Future validation with standardised testing protocols would help integration into clinical care., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://atm.amegroups.com/article/view/10.21037/atm-23-1572/coif). The series “Lung Cancer Management—The Next Decade” was commissioned by the editorial office without any funding or sponsorship. W.K.J.L. has filed multiple patents on cell-free DNA analyses for cancer diagnostics. W.K.J.L. is a member of the Data Advisory Committee of the Hong Kong Genome Institute. W.K.J.L. is a Board member of DRA and hold equities in Illumina. P.J. holds equities in Grail/Illumina, KingMed Future and Take2. PJ is a consultant to KingMed Future and receives consulting fees. P.J. is Director of DRA, Take2, KingMed Future, and Insighta. P.J. has filed various patents or patent applications related to cell-free DNA and receives royalties from Illumina, Take2, DRA, and KingMed Future. The authors have no other conflicts of interest to declare., (2024 Annals of Translational Medicine. All rights reserved.)

Hypochlorite (ClO - ) and viscosity both affect the physiological state of mitochondria, and their abnormal levels are closely related to many common diseases. Therefore, it is vitally important to develop mitochondria-targeting fluorescent probes for the dual sensing of ClO - and viscosity. Herein, we have explored a new fluorescent probe, XTAP - Bn , which responds sensitively to ClO - and viscosity with off-on fluorescence changes at 558 and 765 nm, respectively. Because the emission wavelength gap is more than 200 nm, XTAP - Bn can effectively eliminate the signal crosstalk during the simultaneous detection of ClO - and viscosity. In addition, XTAP - Bn has several advantages, including high selectivity, rapid response, good water solubility, low cytotoxicity, and excellent mitochondrial-targeting ability. More importantly, probe XTAP - Bn is successfully employed to monitor the dynamic change in ClO - and viscosity levels in the mitochondria of living cells and zebrafish. This study not only provides a reliable tool for identifying mitochondrial dysfunction but also offers a potential approach for the early diagnosis of mitochondrial-related diseases.

The phytohormone cytokinin has various roles in plant development, including meristem maintenance, vascular differentiation, leaf senescence, and regeneration. Prior investigations have revealed that cytokinin acts via a phosphorelay similar to the two-component system by which bacteria sense and respond to external stimuli. The eventual targets of this phosphorelay are type-B ARABIDOPSIS RESPONSE REGULATORS (B-ARRs), containing the conserved N-terminal receiver domain (RD), middle DNA binding domain (DBD), and C-terminal transactivation domain. While it has been established for two decades that the phosphoryl transfer from a specific histidyl residue in ARABIDOPSIS HIS PHOSPHOTRANSFER PROTEINS (AHPs) to an aspartyl residue in the RD of B-ARRs results in a rapid transcriptional response to cytokinin, the underlying molecular basis remains unclear. In this work, we determine the crystal structures of the RD-DBD of ARR1 (ARR1 RD-DBD ) as well as the ARR1 DBD -DNA complex from Arabidopsis . Analyses of the ARR1 DBD -DNA complex have revealed the structural basis for sequence-specific recognition of the GAT trinucleotide by ARR1. In particular, comparing the ARR1 RD-DBD and ARR1 DBD -DNA structures reveals that unphosphorylated ARR1 RD-DBD exists in a closed conformation with extensive contacts between the RD and DBD. In vitro and vivo functional assays have further suggested that phosphorylation of the RD weakens its interaction with DBD, subsequently permits the DNA binding capacity of DBD, and promotes the transcriptional activity of ARR1. Our findings thus provide mechanistic insights into phosphorelay activation of gene transcription in response to cytokinin., Competing Interests: Competing interests statement:The authors declare no competing interest.

Transcatheter aortic valve replacement (TAVR) has become the standard of care for high surgical risk patients with aortic stenosis. The most common approach to TAVR is transfemoral utilizing monitored anesthesia care or general anesthesia. On occasion, transfemoral access is not possible and alternative access to TAVR is required. Herein, we describe the case of a patient undergoing a transcarotid approach to TAVR with regional anesthesia and monitored anesthesia care utilizing a multidisciplinary heart team.

Background: Lipid metabolism is a complex process that includes lipid uptake, transport, synthesis, and degradation. Trace elements are vital in maintaining normal lipid metabolism in the human body. This study explores the relationship between serum trace elements and lipid metabolism., Methods: In this study, we reviewed articles on the relationship between alterations in somatic levels of zinc, iron, calcium, copper, chrome, manganese, selenium, and lipid metabolism. In this systematic review and mate-analysis, databases such as PubMed, Web of Science, and China National Knowledge Infrastructure (CNKI), Wanfang was searched for articles on the relationship published between January 1, 1900, and July 12, 2022. The meta-analysis was performed using Review Manager5.3 (Cochrane Collaboration)., Results: No significant association was found between serum zinc and dyslipidemia, while other serum trace elements (iron, selenium, copper, chromium, and manganese) were associated with hyperlipidemia., Conclusion: The present study suggested that the human body's zinc, copper, and calcium content may be related to lipid metabolism. However, findings on lipid metabolism and Iron, Manganese have not been conclusive. In addition, the relationship between lipid metabolism disorders and selenium levels still needs to be further studied. Further research is needed on treating lipid metabolism diseases by changing trace elements., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Protein-biomolecule interactions play pivotal roles in almost all biological processes. For a biomolecule of interest, the identification of the interacting protein(s) is essential. For this need, although many assays are available, highly robust and reliable methods are always desired. By combining a substrate-based proximity labeling activity from the pupylation pathway of Mycobacterium tuberculosis and the streptavidin (SA)-biotin system, we developed the Specific Pupylation as IDEntity Reporter (SPIDER) method for identifying protein-biomolecule interactions. Using SPIDER, we validated the interactions between the known binding proteins of protein, DNA, RNA, and small molecule. We successfully applied SPIDER to construct the global protein interactome for m 6 A and mRNA, identified a variety of uncharacterized m 6 A binding proteins, and validated SRSF7 as a potential m 6 A reader. We globally identified the binding proteins for lenalidomide and CobB. Moreover, we identified SARS-CoV-2-specific receptors on the cell membrane. Overall, SPIDER is powerful and highly accessible for the study of protein-biomolecule interactions., (© 2023. Science China Press.)

Thirteen consecutive patients with entrapped temporal horn syndrome in the Department of Neurosurgery of Beijing Tiantan Hospital from February 2018 to September 2022 were retrospectively analyzed, and there were 5 males and 8 females, with a mean age of (43±21) years. Increased intracranial pressure caused by hydrocephalus was the main clinical symptom. All the patients underwent refined temporal-to-frontal horn shunt, and all the symptoms were improved after surgery. Postoperative Karnofsky performance score (KPS) [90 (90, 100)] was higher than preoperative KPS [57 (40, 70)] ( P =0.001). However, postoperative entrapped temporal horn volume [13.85 (8.90, 15.25) cm 3 ] decreased, compared with preoperative volume [66.52 (38.65, 88.65) cm 3 ] ( P =0.001). Likewise, postoperative midline shift [0.77 (0, 1.50) mm] was longer than preoperative midline shift [6.69 (2.50, 10.00) mm] ( P =0.002). No surgery-related complications were observed after the operation. Therefore, the refined temporal-to-frontal horn shunt is safe and effective treatment for entrapped temporal horn syndrome, with favorable outcomes.

Cell-free DNA (cfDNA) fragmentation is nonrandom, at least partially mediated by various DNA nucleases, forming characteristic cfDNA end motifs. However, there is a paucity of tools for deciphering the relative contributions of cfDNA cleavage patterns related to underlying fragmentation factors. In this study, through non-negative matrix factorization algorithm, we used 256 5' 4-mer end motifs to identify distinct types of cfDNA cleavage patterns, referred to as "founder" end-motif profiles (F-profiles). F-profiles were associated with different DNA nucleases based on whether such patterns were disrupted in nuclease-knockout mouse models. Contributions of individual F-profiles in a cfDNA sample could be determined by deconvolutional analysis. We analyzed 93 murine cfDNA samples of different nuclease-deficient mice and identified six types of F-profiles. F-profiles I, II, and III were linked to deoxyribonuclease 1 like 3 (DNASE1L3), deoxyribonuclease 1 (DNASE1), and DNA fragmentation factor subunit beta (DFFB), respectively. We revealed that 42.9% of plasma cfDNA molecules were attributed to DNASE1L3-mediated fragmentation, whereas 43.4% of urinary cfDNA molecules involved DNASE1-mediated fragmentation. We further demonstrated that the relative contributions of F-profiles were useful to inform pathological states, such as autoimmune disorders and cancer. Among the six F-profiles, the use of F-profile I could inform the human patients with systemic lupus erythematosus. F-profile VI could be used to detect individuals with hepatocellular carcinoma, with an area under the receiver operating characteristic curve of 0.97. F-profile VI was more prominent in patients with nasopharyngeal carcinoma undergoing chemoradiotherapy. We proposed that this profile might be related to oxidative stress.

The data of 1 268 newly diagnosed gliomas from the Fourth Ward of Neurosurgery Department of Beijing Tiantan Hospital between April 2013 and March 2022 were retrospectively analyzed. Based on postoperative pathology, the gliomas were divided into groups: oligodendrogliomas ( n =308), astrocytomas ( n =337) and glioblastomas ( n =623). According to the O6-methylguanine-DNA methyl transferase (MGMT) promoter status defined by the 12% of best cut-off value in previous research results, patients were divided into methylation group ( n =763) and non-methylation group ( n =505). Methylation level [ M ( Q 1 , Q 3 )] in patients with glioblastoma, astrocytoma and oligodendroglioma was 6% (2%, 24%), 17% (10%, 28%) and 29% (19%, 40%), respectively ( P <0.001). Compared with non-methylation patients, the progression-free survival (PFS) and overall survival (OS) of glioblastomas patients with methylation of MGMT promoter demonstrated more favorable prognosis [ M ( Q 1 , Q 3 )]) of PFS: 14.0 (6.0, 36.0) months vs 8.0 (4.0, 15.0) months, P <0.001; M ( Q 1 , Q 3 ) of OS: 29.0 (17.0, 60.5) months vs 16.0 (11.0, 26.5) months, P <0.001]. In astrocytomas patients, the PFS was much longer for those with methylation [the median PFS of patients with methylation was not observed at the end of follow-up, but those without methylation showed a median PFS of 46.0 (29.0, 52.0) months] ( P =0.001). However, no statistically significant difference was observed in OS [the median OS of patients with methylation was not observed at the end of follow-up, but those without methylation had a median OS of 62.0 (46.0, 98.0) months] ( P =0.085). In oligodendrogliomas patients, no statistically significant differences of PFS and OS were observed between patients with methylation and those without methylation. MGMT promoter status was a related factor affecting PFS and OS in glioblastomas (PFS: HR =0.534, 95%CI : 0.426-0.668, P <0.001; OS: HR =0.451, 95%CI : 0.353-0.576, P <0.001). Moreover, MGMT promoter status was also a related factor affecting PFS in astrocytomas ( HR =0.462, 95% CI : 0.221-0.966, P =0.040), but not for OS ( HR =0.664, 95% CI : 0.259-1.690, P =0.389). The methylation level of MGMT promoter differed substantially in different types of gliomas, and the status of MGMT promoter profoundly affected the prognosis of glioblastomas.

Background: Nuclear-derived cell-free DNA (cfDNA) molecules in blood plasma are nonrandomly fragmented, bearing a wealth of information related to tissues of origin. DNASE1L3 (deoxyribonuclease 1 like 3) is an important player in shaping the fragmentation of nuclear-derived cfDNA molecules, preferentially generating molecules with 5 CC dinucleotide termini (i.e., 5 CC-end motif). However, the fragment end properties of microbial cfDNA and its clinical implication remain to be explored., Methods: We performed end motif analysis on microbial cfDNA fragments in plasma samples from patients with sepsis. A sequence context-based normalization method was used to minimize the potential biases for end motif analysis., Results: The end motif profiles of microbial cfDNA appeared to resemble that of nuclear cfDNA (Spearman correlation coefficient: 0.82, P value 0.001). The CC-end motif was the most preferred end motif in microbial cfDNA, suggesting that DNASE1L3 might also play a role in the fragmentation of microbe-derived cfDNA in plasma. Of note, differential end motifs were present between microbial cfDNA originating from infection-causing pathogens (enriched at the CC-end) and contaminating microbial DNA potentially derived from reagents or the environment (nearly random). The use of fragment end signatures allowed differentiation between confirmed pathogens and contaminating microbes, with an area under the receiver operating characteristic curve of 0.99. The performance appeared to be superior to conventional analysis based on microbial cfDNA abundance alone., Conclusions: The use of fragmentomic features could facilitate the differentiation of underlying contaminating microbes from true pathogens in sepsis. This work demonstrates the potential usefulness of microbial cfDNA fragmentomics in metagenomics analysis., (© American Association for Clinical Chemistry 2022.)

The majority of people in China have been immunized with the inactivated viral vaccine BBIBP-CorV. The emergence of the Omicron variant raised the concerns about protection efficacy of the inactivated viral vaccine in China. However, longitudinal neutralization data describing protection efficacy against Omicron variant is still lacking. Here we present one-year longitudinal neutralization data of BBIBP-CorV on authentic Omicron, Delta, and wild-type strains using 224 sera collected from 14 volunteers who have finished three doses BBIBP-CorV. The sera were also subjected for monitoring the SARS-CoV-2 specific IgG, IgA, and IgM responses on protein and peptide microarrays. The neutralization titers showed different protection efficacies against the three strains. By incorporating IgG and IgA signals of proteins and Spike protein derived peptide on microarray, panels as potential surrogate biomarkers for rapid estimation of neutralization titers were established. These data support the necessity of the 3 rd dose of BBIBP-CorV vaccination. After further validation and assay development, the panels could be used for reliable, convenient and fast evaluation of the efficacy of vaccination., (© 2022 Wiley-VCH GmbH.)

Rod-like particles have attracted increasing attention because of their unique shape-dependent properties, which enable their superior performance compared to their isotropic counterparts. Thus, rod-like particles have potential applications in many fields, especially in biomedicine. However, the fabrication of uniform rod-like particles is challenging because of the principle of interfacial energy minimization. Herein, we present a facile, rapid, and cost-effective strategy for preparing starch-based microrods with tunable aspect ratios via shear-assisted antisolvent-induced nanoprecipitation and solidification. The preformed spherical particles swollen by the mixed solvent were elongated by the shear force and solidified in rod-like shape by antisolvent induction. The resulting starch-based microrods can encapsulate hydrophobic active substances and be modified with functional groups, indicating their potential applications as drug carriers and biologically active materials. The formation mechanism of the starch-based microrods discovered in this study provides a new perspective on the fabrication of rod-like polymer particles.

Cell-free DNA (cfDNA) fragmentation patterns contain important molecular information linked to tissues of origin. We explored the possibility of using fragmentation patterns to predict cytosine-phosphate-guanine (CpG) methylation of cfDNA, obviating the use of bisulfite treatment and associated risks of DNA degradation. This study investigated the cfDNA cleavage profile surrounding a CpG (i.e., within an 11-nucleotide [nt] window) to analyze cfDNA methylation. The cfDNA cleavage proportion across positions within the window appeared nonrandom and exhibited correlation with methylation status. The mean cleavage proportion was ∼twofold higher at the cytosine of methylated CpGs than unmethylated ones in healthy controls. In contrast, the mean cleavage proportion rapidly decreased at the 1-nt position immediately preceding methylated CpGs. Such differential cleavages resulted in a characteristic change in relative presentations of CGN and NCG motifs at 5' ends, where N represented any nucleotide. CGN/NCG motif ratios were correlated with methylation levels at tissue-specific methylated CpGs (e.g., placenta or liver) (Pearson's absolute r > 0.86). cfDNA cleavage profiles were thus informative for cfDNA methylation and tissue-of-origin analyses. Using CG-containing end motifs, we achieved an area under a receiver operating characteristic curve (AUC) of 0.98 in differentiating patients with and without hepatocellular carcinoma and enhanced the positive predictive value of nasopharyngeal carcinoma screening (from 19.6 to 26.8%). Furthermore, we elucidated the feasibility of using cfDNA cleavage patterns to deduce CpG methylation at single CpG resolution using a deep learning algorithm and achieved an AUC of 0.93. FRAGmentomics-based Methylation Analysis (FRAGMA) presents many possibilities for noninvasive prenatal, cancer, and organ transplantation assessment.

Background: Currently, there are many therapeutic methods for lung adenocarcinoma (LUAD), but the 5-year survival rate is still only 15% at later stages. Epithelial- mesenchymal transition (EMT) has been shown to be closely associated with local dissemination and subsequent metastasis of solid tumors. However, the role of EMT in the occurrence and development of LUAD remains unclear., Aim: To further elucidate the value of EMT-related genes in LUAD prognosis., Methods: Univariate, least absolute shrinkage and selection operator, and multivariate Cox regression analyses were applied to establish and validate a new EMT-related gene signature for predicting LUAD prognosis. The risk model was evaluated by Kaplan-Meier survival analysis, principal component analysis, and functional enrichment analysis and was used for nomogram construction. The potential structures of drugs to which LUAD is sensitive were discussed with respect to EMT-related genes in this model., Results: Thirty-three differentially expressed genes related to EMT were found to be highly associated with overall survival (OS) by using univariate Cox regression analysis (log2FC ≥ 1, false discovery rate < 0.001). A prognostic signature of 7 EMT-associated genes was developed to divide patients into two risk groups by high or low risk scores. Kaplan-Meier survival analysis showed that the OS of patients in the high-risk group was significantly poorer than that of patients in the low-risk group ( P < 0.05). Multivariate Cox regression analysis showed that the risk score was an independent risk factor for OS (HR > 1, P < 0.05). The results of receiver operator characteristic curve analysis suggested that the 7-gene signature had a perfect ability to predict prognosis (all area under the curves > 0.5)., Conclusion: The EMT-associated gene signature classifier could be used as a feasible indicator for predicting OS., Competing Interests: Conflict-of-interest statement: The authors declare no conflicts of interest., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

A better knowledge of the spatial and temporal variation in atmospheric aerosol and its influencing factors is of great significance to controlling atmospheric pollution and improving the atmospheric environment. First, the visible infrared imaging radiometer suite (VIIRS) intermediate product (IP) aerosol optical depth (AOD) data from 2013 to 2019 were used to analyze the temporal and spatial variation in AOD in the North China Plain. Secondly, SO 2 , NO 2 , PM 2.5 , meteorological data, NDVI, DEM, GDP, and POPU were selected as influencing factors, and the linkage models between AOD and its influencing factors were established based on the XGBoost model for each of the five representative cities in the North China Plain to quantitatively estimate and reveal the contribution of various influencing factors behind the temporal and spatial distribution in AOD. The results showed that in terms of spatial distribution, the AOD of the North China Plain was bounded by the Taihang Mountains, showing a pattern of high AOD in the southeast and low AOD in the northwest. In terms of temporal changes, the annual average value of AOD in the five cities showed an overall decreasing trend, and the monthly average value of AOD first increased and then decreased, with the highest value appearing in July and the lowest value in December. In addition, the AOD estimation model established in this paper for the five cities in North China had high accuracy, with R 2 ranging from 0.60 to 0.67. Among the factors influencing AOD in the North China Plain, NO 2 and SO 2 were the most influential factors contributing to AOD in the five cities. In addition, PM 2.5 was another important pollutant emission factor. In terms of meteorological factors, temperature ( T ), relative humidity (RH), wind speed (WS), and wind direction (WD) were the other four important influencing factors. There were both commonalities and differences in the rankings of the contribution and importance of AOD influencing factors in the five representative cities in North China.

Urinary cell-free DNA (ucfDNA) is a potential biomarker for bladder cancer detection. However, the biological characteristics of ucfDNA are not well understood. We explored the roles of deoxyribonuclease 1 (DNASE1) and deoxyribonuclease 1-like 3 (DNASE1L3) in the fragmentation of ucfDNA using mouse models. The deletion of Dnase1 in mice (Dnase1-/-) caused aberrations in ucfDNA fragmentation, including a 24-fold increase in DNA concentration, and a 3-fold enrichment of long DNA molecules, with a relative decrease of fragments with thymine ends and reduction of jaggedness (i.e., the presence of single-stranded protruding ends). In contrast, such changes were not observed in mice with Dnase1l3 deletion (Dnase1l3-/-). These results suggested that DNASE1 was an important nuclease contributing to the ucfDNA fragmentation. Western blot analysis revealed that the concentration of DNASE1 protein was higher in urine than DNASE1L3. The native-polyacrylamide gel electrophoresis zymogram showed that DNASE1 activity in urine was higher than that in plasma. Furthermore, the proportion of ucfDNA fragment ends within DNase I hypersensitive sites (DHSs) was significantly increased in Dnase1-deficient mice. In humans, patients with bladder cancer had lower proportions of ucfDNA fragment ends within the DHSs when compared with participants without bladder cancer. The area under the curve (AUC) for differentiating patients with and without bladder cancer was 0.83, suggesting the analysis of ucfDNA fragmentation in the DHSs may have potential for bladder cancer detection. This work revealed the intrinsic links between the nucleases in urine and ucfDNA fragmentomics., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: K.C.A.C., R.W.K.C., and Y.M.D.L. hold equities in DRA, Take2, and Grail/Illumina. K.C.A.C., R.W.K.C., and Y.M.D.L. were consultants to Grail/Illumina. K.C.A.C., R.W.K.C., and Y.M.D.L. received research funding from Grail/Illumina. Y.M.D.L. was a scientific co-founder of and served on the scientific advisory board of Grail/Illumina. W.K.J.L and P.J. holds equities in Grail/Illumina. P.J. is a consultant to Take2. P.J. is a Director of KingMed Future.

Background: Posterior internal fixation (PIF) is commonly used in the treatment of thoracolumbar fracture (TLF), but there is still no standard for the number of fixed segments. The objective of this meta-analysis was to evaluate the efficacy and safety of short segment (SS), intermediate segment (IS) and long segment (LS) in the fixation of TLF., Methods: Two authors independently searched through PubMed, Embase, Cochrane Library and Web of Science for studies of thoracolumbar fracture treated by posterior internal fixation, which were published until the end of April 2021. The Aggregate Data Drug Information System (ADDIS) software was used for data evaluation according to the Markov chain Monte Carlo (MCMC) method based on the Bayesian theorem., Results: Nineteen trials evaluating a total of 970 patients were enrolled in these studies, of which 340 in the SS group, 429 in the IS group and 201 in the LS group. For anterior vertebral height ratio (AVHR), IS had the highest AVHR, LS had the second highest AVHR. IS also ranked first in reducing visual analogue scale (VAS), SS ranked second. For sagittal Cobb's angle (SCA), LS had the lowest SCA and IS had the second lowest SCA. In terms of adverse events, IS had the lowest implant failure rate and LS had the second lowest implant failure rate., Conclusions: IS may be the most desirable treatment option for TLF in reducing SCA, implant failure rate, VAS, and improving AVHR. However, more randomized controlled trials are needed to verify these results., Competing Interests: Declaration of Competing Interest None., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Major depressive disorder is a serious and common neuropsychiatric disorder that affects more than 350 million people worldwide. Electroconvulsive therapy is the oldest and most effective treatment available for the treatment of severe major depressive disorder. Electroconvulsive therapy modifies structural network changes in patients with major depressive disorder and schizophrenia. And it can also affect neuroinflammatory responses and may have neuroprotective effects. Electroconvulsive therapy plays an irreplaceable role in the treatment of major depressive disorder., Competing Interests: Conflict-of-interest statement: The authors declare no potential conflicts of interest with respect to the research, authorship, or publication of this article., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Pharmacological inhibitors of glutathione synthesis and circulation, such as buthionine-sulfoximine, inhibit glutathione metabolism. These drugs decrease the aggressiveness of pancreatic cancer, inhibit tumor stem cell survival, and reduce chemotherapy resistance. Nevertheless, buthionine-sulfoximine also decreases the content of glutathione in normal cells, disrupts the balance between reactive oxygen species and glutathione, and eventually induces cell apoptosis. Pancreatic cancer is usually diagnosed at an advanced stage and has a poor prognosis. Consequently, the use of biomarkers to screen high-risk patients can be an effective method., Competing Interests: Conflict-of-interest statement: The authors declare that they have no conflict of interest., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Epilepsy (EP) is a common neurological disorder which is characterized by excessive abnormal synchronization of neuronal discharges in the brain due to chronic recurrent seizures of multiple etiologies. Variety of microRNAs have been associated with the occurrence and development of EP. This study aimed to determine the aberrant expression of miR-378 and miR-575 in EP patients to validate their potential to distinguish EP from healthy patients., Methods: RT-qPCR was used to determine the expressions of miR-378 and miR-575 from serum specimens of 106 EP and 103 control individuals. Clinical indicators between EP patients and controls were assessed. Based on surgical outcome, EP patients were further divided into Engel I-IV EP. The potentials of miR-378 and miR-575 in discriminating EP from healthy participants and predicting surgical prognosis were calculated by receiver operating characteristic (ROC) analysis., Results: We found the miR-378 and miR-575 were significantly declined (P<0.001) in Engel I-II and III-IV EP patients with no difference in clinical parameters compared. Moreover, miR-378 and miR-575 displayed high sensitivity, specificity, and accuracy in distinguishing EP patients and predicting surgical outcomes. Moreover, after surgical treatment, miR-378 and miR-575 levels were increased compared with those at admission, suggesting their potentials in treatment response., Conclusions: miR-378 and miR-575 could be utilized as novel and non-invasive serum biomarkers in discriminating EP from healthy controls and predicting surgical outcome, shedding new insights on epileptogenesis and EP treatment., Competing Interests: Conflict of Interest: The authors stated that they have no conflicts of interest regarding the publication of this article., (2022 Xiuxiu Li, Zhiqing Gao, Mei Ling Ma, Li Li, Shifeng Guo, published by CEON/CEES.)

There are many causes of anemia. It is unreasonable to simply associate anemia with asymptomatic ulcers in the small intestine., Competing Interests: Conflict-of-interest statement: Each author stated that there is no conflict of interest., (©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.)

Single-stranded ends of double-stranded DNA (jagged ends) are more abundant in urinary DNA than in plasma DNA. However, the lengths of jagged ends in urinary DNA remained undetermined, as a previous method used for urinary DNA jagged end sequencing analysis (Jag-seq) relied on unmethylation at CpG sites, limiting the resolution. Here, we performed high-resolution Jag-seq analysis using methylation at non-CpG cytosine sites, allowing determination of exact length of jagged ends. The urinary DNA bore longer jagged ends (~26-nt) than plasma DNA (~17-nt). The jagged end length distribution displayed 10-nt periodicities in urinary DNA, which were much less observable in plasma DNA. Amplitude of the 10-nt periodicities increased in patients with renal cell carcinoma. Heparin treatment of urine diminished the 10-nt periodicities. The urinary DNA jagged ends often extended into nucleosomal cores, suggesting potential interactions with histones. This study has thus advanced our knowledge of jagged ends in urine DNA., (© 2022. The Author(s).)

The purpose of this study was to compare the clinical effectiveness of minimally invasive clamp-assisted reduction and open reduction with wire cerclage and intramedullary nails for unstable subtrochanteric fractures. Between January 2016 and October 2019, 68 patients who had unstable subtrochanteric fractures experienced intramedullary nail surgery in this retrospective study. There were 41 cases in the minimally invasive clamp or closed reduction group (group A) and 27 cases in the open reduction with wire cerclage group (group B). There were 3 cases of complications in group A and 2 cases of complications in group B. Remarkable distinction was observed between the two groups in the operation time ( p < 0.05), quality of reduction ( p < 0.05), and union time ( p < 0.05). For the successful surgical treatment of unstable femoral subtrochanteric fractures, an anatomical reduction is crucial. Reduction and wire cerclage are cut to give medial support for the anatomical reduction, which has a positive effect on fracture healing., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this study., (Copyright © 2022 Dong Liu et al.)

Introduction: To improve compliance with voiding diaries in children with primary monosymptomatic nocturnal enuresis (PMNE), a new modified 3-day weekend frequency-volume chart (FVC) was designed, and the compliance and validity of this modified FVC was evaluated by comparing with the International Children's Continence Society (ICCS) recommended voiding diary., Methods: A total of 1200 patients with PMNE were enrolled in the study from 13 centers in China and were randomly assigned to record this modified FVC or the ICCS-recommended voiding diary. The primary outcome measure was the compliance, assessed by comparing the completing index and the quality score of diaries between two groups. The secondary outcome measure was the validity, evaluated by comparing the constituent of subtypes, micturition parameters and response rate to desmopressin., Results: Among the 1200 participants enrolled in the study, 447 patients completed the ICCS-recommended voiding diary and 469 completed the modified diary. The diurnal completing index and the quality score of the modified FVC group were better than those of the ICCS group. In addition, there was no significant difference between these two groups in the subtype classification, or in the response rate to desmopressin., Conclusions: The modified FVC could be applied to obtain the voiding characteristics of children with PMNE as the ICCS-recommended voiding diary does and offers a reasonable and better choice for children with PMNE from the unselected population in the future., (© 2021. Children's Hospital, Zhejiang University School of Medicine.)

The etiology of psychiatric disorders remains largely unknown. The exploration of the neurobiological mechanisms of mental illness helps improve diagnostic efficacy and develop new therapies. This review focuses on the application of concurrent transcranial magnetic stimulation and electroencephalography (TMS-EEG) in various mental diseases, including major depressive disorder, bipolar disorder, schizophrenia, autism spectrum disorder, attention-deficit/hyperactivity disorder, substance use disorder, and insomnia. First, we summarize the commonly used protocols and output measures of TMS-EEG; then, we review the literature exploring the alterations in neural patterns, particularly cortical excitability, plasticity, and connectivity alterations, and studies that predict treatment responses and clinical states in mental disorders using TMS-EEG. Finally, we discuss the potential mechanisms underlying TMS-EEG in establishing biomarkers for psychiatric disorders and future research directions. This article is part of the special Issue on 'Stress, Addiction and Plasticity'., (Copyright © 2021. Published by Elsevier Ltd.)

Long noncoding RNA (lncRNA) CDKN2B‑antisense RNA 1 (AS1) functions as a tumor oncogene in numerous cancers. However, the roles and mechanism of CDKN2B‑AS1 in colorectal cancer (CRC) have not been explored. The present study aimed to investigate whether and how CDKN2B‑AS1 contributes to CRC progression. The data revealed that CDKN2B‑AS1 expression was upregulated in CRC tissues. Loss‑of‑function assays demonstrated that CDKN2B‑AS1 in CRC modulated cell proliferation and apoptosis, which was mediated by cyclin D1, cyclin‑dependent kinase (CDK) 4, p‑Rb, caspase‑9 and caspase‑3. Bioinformatics analysis and luciferase reporter assays indicated direct binding of microRNA (miR)‑28‑5p to CDKN2B‑AS1. Moreover, the results herein revealed that the expression of miR‑28‑5p was negatively correlated with that of CDKN2B‑AS1 in CRC tissue. Moreover, CDKN2B‑AS1 acted as a miR‑28‑5p competing endogenous RNA (ceRNA) to target and regulate the expression of URGCP. These findings indicated that CDKN2B‑AS1 plays roles in CRC progression, providing a potential therapeutic target or novel diagnostic biomarker for CRC.

Coronavirus disease 2019 (COVID-19), which is caused by SARS-CoV-2, varies with regard to symptoms and mortality rates among populations. Humoral immunity plays critical roles in SARS-CoV-2 infection and recovery from COVID-19. However, differences in immune responses and clinical features among COVID-19 patients remain largely unknown. Here, we report a database for COVID-19-specific IgG/IgM immune responses and clinical parameters (named COVID-ONE-hi). COVID-ONE-hi is based on the data that contain the IgG/IgM responses to 24 full-length/truncated proteins corresponding to 20 of 28 known SARS-CoV-2 proteins and 199 spike protein peptides against 2360 serum samples collected from 783 COVID-19 patients. In addition, 96 clinical parameters for the 2360 serum samples and basic information for the 783 patients are integrated into the database. Furthermore, COVID-ONE-hi provides a dashboard for defining samples and a one-click analysis pipeline for a single group or paired groups. A set of samples of interest is easily defined by adjusting the scale bars of a variety of parameters. After the "START" button is clicked, one can readily obtain a comprehensive analysis report for further interpretation. COVID-ONE-hi is freely available at www.COVID-ONE.cn., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

One of the best ways to control COVID-19 is vaccination. Among the various SARS-CoV-2 vaccines, inactivated virus vaccines have been widely applied in China and many other countries. To understand the underlying protective mechanism of these vaccines, it is necessary to systematically analyze the humoral responses that are triggered. By utilizing a SARS-CoV-2 microarray with 21 proteins and 197 peptides that fully cover the spike protein, antibody response profiles of 59 serum samples collected from 32 volunteers immunized with the inactivated virus vaccine BBIBP-CorV were generated. For this set of samples, the microarray results correlated with the neutralization titers of the authentic virus, and two peptides (S1-5 and S2-22) were identified as potential biomarkers for assessing the effectiveness of vaccination. Moreover, by comparing immunized volunteers to convalescent and hospitalized COVID-19 patients, the N protein, NSP7, and S2-78 were identified as potential biomarkers for differentiating COVID-19 patients from individuals vaccinated with the inactivated SARS-CoV-2 vaccine. The comprehensive profile of humoral responses against the inactivated SARS-CoV-2 vaccine will facilitate a deeper understanding of the vaccine and provide potential biomarkers for inactivated virus vaccine-related applications., (© 2021. The Author(s).)

The immunogenicity of the SARS-CoV-2 proteome is largely unknown, especially for non-structural proteins and accessory proteins. In this study, we collect 2,360 COVID-19 sera and 601 control sera. We analyze these sera on a protein microarray with 20 proteins of SARS-CoV-2, building an antibody response landscape for immunoglobulin (Ig)G and IgM. Non-structural proteins and accessory proteins NSP1, NSP7, NSP8, RdRp, ORF3b, and ORF9b elicit prevalent IgG responses. The IgG patterns and dynamics of non-structural/accessory proteins are different from those of the S and N proteins. The IgG responses against these six proteins are associated with disease severity and clinical outcome, and they decline sharply about 20 days after symptom onset. In non-survivors, a sharp decrease of IgG antibodies against S1 and N proteins before death is observed. The global antibody responses to non-structural/accessory proteins revealed here may facilitate a deeper understanding of SARS-CoV-2 immunology., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

To fully decipher the immunogenicity of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Spike protein, it is essential to assess which part is highly immunogenic in a systematic way. We generate a linear epitope landscape of the Spike protein by analyzing the serum immunoglobulin G (IgG) response of 1,051 coronavirus disease 2019 (COVID-19) patients with a peptide microarray. We reveal two regions rich in linear epitopes, i.e., C-terminal domain (CTD) and a region close to the S2' cleavage site and fusion peptide. Unexpectedly, we find that the receptor binding domain (RBD) lacks linear epitope. We reveal that the number of responsive peptides is highly variable among patients and correlates with disease severity. Some peptides are moderately associated with severity and clinical outcome. By immunizing mice, we obtain linear-epitope-specific antibodies; however, no significant neutralizing activity against the authentic virus is observed for these antibodies. This landscape will facilitate our understanding of SARS-CoV-2-specific humoral responses and might be useful for vaccine refinement., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Serological tests play an essential role in monitoring and combating the COVID-19 pandemic. Recombinant spike protein (S protein), especially the S1 protein, is one of the major reagents used for serological tests. However, the high cost of S protein production and possible cross-reactivity with other human coronaviruses pose unavoidable challenges. By taking advantage of a peptide microarray with full spike protein coverage, we analyzed 2,434 sera from 858 COVID-19 patients, 63 asymptomatic patients and 610 controls collected from multiple clinical centers. Based on the results, we identified several S protein-derived 12-mer peptides that have high diagnostic performance. In particular, for monitoring the IgG response, one peptide (aa 1148-1159 or S2-78) exhibited a sensitivity (95.5%, 95% CI 93.7-96.9%) and specificity (96.7%, 95% CI 94.8-98.0%) comparable to those of the S1 protein for the detection of both symptomatic and asymptomatic COVID-19 cases. Furthermore, the diagnostic performance of the S2-78 (aa 1148-1159) IgG was successfully validated by ELISA in an independent sample cohort. A panel of four peptides, S1-93 (aa 553-564), S1-97 (aa 577-588), S1-101 (aa 601-612) and S1-105 (aa 625-636), that likely will avoid potential cross-reactivity with sera from patients infected by other coronaviruses was constructed. The peptides identified in this study may be applied independently or in combination with the S1 protein for accurate, affordable, and accessible COVID-19 diagnosis.

Objectives: Due to the maternally-inherited nature of mitochondrial DNA (mtDNA), there is a lack of information regarding fetal mtDNA in the plasma of pregnant women. We aim to explore the presence and topologic forms of circulating fetal and maternal mtDNA molecules in surrogate pregnancies., Methods: Genotypic differences between fetal and surrogate maternal mtDNA were used to identify the fetal and maternal mtDNA molecules in plasma. Plasma samples were obtained from the surrogate pregnant mothers. Using cleavage-end signatures of BfaI restriction enzyme, linear and circular mtDNA molecules in maternal plasma could be differentiated., Results: Fetal-derived mtDNA molecules were mainly linear (median: 88%; range: 80%-96%), whereas approximately half of the maternal-derived mtDNA molecules were circular (median: 51%; range: 42%-60%). The fetal DNA fraction of linear mtDNA was lower (median absolute difference: 9.8%; range: 1.1%-27%) than that of nuclear DNA (median: 20%; range: 9.7%-35%). The fetal-derived linear mtDNA molecules were shorter than the maternal-derived ones., Conclusion: Fetal mtDNA is present in maternal plasma, and consists mainly of linear molecules. Surrogate pregnancies represent a valuable clinical scenario for exploring the biology and potential clinical applications of circulating mtDNA, for example, for pregnancies conceived following mitochondrial replacement therapy., (© 2020 The Authors. Prenatal Diagnosis published by John Wiley & Sons Ltd.)

The adhesion GPCR ADGRG2, also known as GPR64, is a critical regulator of male fertility that maintains ion/pH homeostasis and CFTR coupling. The molecular basis of ADGRG2 function is poorly understood, in part because no endogenous ligands for ADGRG2 have been reported, thus limiting the tools available to interrogate ADGRG2 activity. It has been shown that ADGRG2 can be activated by a peptide, termed p15, derived from its own N-terminal region known as the Stachel sequence. However, the low affinity of p15 limits its utility for ADGRG2 characterization. In the current study, we used alanine scanning mutagenesis to examine the critical residues responsible for p15-induced ADGRG2 activity. We next designed systematic strategies to optimize the peptide agonist of ADGRG2, using natural and unnatural amino acid substitutions. We obtained an optimized ADGRG2 Stachel peptide T1V/F3Phe(4-Me) (VPM-p15) that activated ADGRG2 with significantly improved (>2 orders of magnitude) affinity. We then characterized the residues in ADGRG2 that were important for ADGRG2 activation in response to VPM-p15 engagement, finding that the toggle switch W 6.53 and residues of the ECL2 region of ADGRG2 are key determinants for VPM-p15 interactions and VPM-p15-induced Gs or arrestin signaling. Our study not only provides a useful tool to investigate the function of ADGRG2 but also offers new insights to guide further optimization of Stachel peptides to activate adhesion GPCR members., Competing Interests: Conflict of interest The authors declare no conflicts of interest in regards to this manuscript., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The specific expression profile and function of circular RNAs (circRNAs) in mammalian ovarian follicles, especially during the atresia process, are unclear. In this study, genome-wide deep circRNA sequencing was applied to screen circRNAs in healthy and early atretic antral follicles in pig ovaries. A total of 40,567 distinct circRNAs were identified in follicles, among which 197 circRNAs (108 upregulated and 89 downregulated) were significantly shifted during the early atresia process. Most differentially expressed circRNAs (DECs) lacked protein-coding potential. Annotation analysis of the DECs revealed 162 known host genes, or noncoding RNAs, and 10 intergenic regions. The key pathways in which these host genes are involved include the focal adhesion-PI3K-Akt-mTOR signaling pathway, vascular endothelial growth factor A (VEGFA)-vascular endothelial growth factor receptor 2 signaling pathway and transforming growth factor-beta signaling pathway. Further comparison analysis between host genes of DECs and the differentially expressed linear messenger RNA transcripts revealed the cotranscription of circRNAs and their linear mRNAs in inhibin beta units (INHBA and INHBB), glutathione S-transferase (GSTA1), and VEGFA. In addition, we predicted 196 pairs of potential circRNA-micro RNA (miRNA) interactions among 77 DECs and 101 porcine miRNAs. We have identified 16 functional miRNAs by comparing the 101 miRNAs to the functional miRNAs reported in mammal ovarian follicle atresia and granulosa cell apoptosis studies. Our study adds new knowledge to circRNA distribution profiles in pig ovarian follicles, offers a valuable reference for transcriptomic profiles in the initiation of follicular atresia, highlights warranted circRNAs for further functional investigation, and provides possible biomarkers for ovarian dysfunctions., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Objective: To analyze the prognosis factors of cerebrospinal fluid (CSF) spread after surgery in glioblastoma (GBM) patients when tumors progressed and the effect factors on prognosis. Methods: A retrospective study was conducted on 124 patients who were pathologically diagnosed as glioblastoma after surgery, and found tumor progressed during regularly follow-up at Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University between January 2009 and August 2017.There were 82 males and 42 females, aged 47.9 years(range: 19 to 75 years) .Patients were divided into local recurrence group(96 cases) and CSF spread group (28 cases) .Clinical data were recorded in detail and compared by independent sample t test or χ(2) test.Kaplan-Meier survival curves was used to demonstrated the distribution of progression free survival (PFS) overall survival (OS) and post progression survival (PPS), and differences between local recurrence and CSF spread groups were assessed by Log-rank test.Cox proportion hazard regression analysis was used to identify independent prognostic factors. Results: Logistics regression analysis showed ventricle entry was the only prognosis factor of CSF spread ( OR= 2.667, 95 % CI : 1.128 to 6.304, P= 0.025).No significant distinction was observed in PFS between CSF spread group and local recurrence group(7.0 months vs .9.3 months, P= 0.066).However, OS and PPS were substantially shortened in CSF spread group (13.0 months vs .23.0 months, P= 0.011; 6.0 months vs .11.0 months, P= 0.022, respectively).Mutations of isocitrate dehydrogenase gene, distant spread, gross-total resection, Ki-67 index>30% were independent prognostic factors of GBM patients. Conclusions: Ventricle entry is a prognosis factor for CSF spread, after which the median OS and PPS are markedly diminished.However, ventricle entry is not independent prognosis factor shortening survival.

Objective: To explore the clinical characteristics and follow-up outcomes of a pedigree of maturity onset diabetes of the young (MODY) induced by a novel mutation of glucokinase (GCK). Methods: The clinical features and laboratory data of a pedigree diagnosed with GCK-MODY in Peking Union Medical College Hospital was analyzed. Genomic DNA was extracted, and Sanger sequencing was performed to detect the gene mutation of the family members. The proband and her father were followed up for 3 years. Wanfang and PubMed were used to search literatures on follow-up studies for treatment of GCK-MOYD. Results: Both the proband and her father were found to have a novel mutation on the GCK gene located in exo10 c.1348G.T (p. Ala450Thr). The proband was treated with diet and exercise control only. At the end of the follow-up, her fasting plasma glucose (FPG, 6.8 mmol/L), 2 h postprandial plasma glucose (2hPG, 7.4 mmol/L), and glycated hemoglobin (HbA1c, 6.3%) were all within the control targets. Additionally, the levels homeostasis model assessment of insulin resistance (HOMA-IR) tended to improved comparing to that at baseline (4.09 to 2.32), and glucose disposition index (DI) was improved compared with baseline (16.22 to 20.05). As to the proband's father, the treatment with insulin plus acarbose was converted to sulfonylureas monotherapy. His FPG and 2hPG mostly were within the target range, and the levels of HbA1c were significantly reduced by 0.5%-0.7% when compared to that at baseline. The HOMA-IR or islet beta cell function was comparable to those at baseline. Conclusions: Screening patients whose clinical performance meets GCK-MODY and their family members with proper genetic testing is of great importance to reduce misdiagnosis of GCK-MODY, so as to obtain a better glucose control without unnecessary over-treatment and protect islet beta cell function.

Background: Laparoscopic common bile duct exploration (LCBDE) is one of the minimally invasive options for choledocholithiasis. Primary closure of the common bile duct (CBD) upon completion of laparoscopic choledochotomy is safe in selected patients. The present study aimed to evaluate the feasibility and safety of primary closure of CBD after LCBDE in patients aged 70 years or older., Methods: A total of 116 patients (51 males and 65 females) who suffered from choledocholithiasis and underwent primary closure of the CBD (without T-tube drainage) after LCBDE from January 2003 to December 2017 were recruited. They were classified into two groups according to age: group A (≥70 years, n = 56), and group B (<70 years, n = 60). The preoperative characteristics, intraoperative details, and postoperative outcomes of the two groups were evaluated., Results: The mean operative time was 172.02 min for group A and 169.92 min for group B (P = 0.853). The mean hospital stay was 7.40 days for group A and 5.38 days for group B (P < 0.001). Bile leakage occurred in two patients in group A and one in group B (3.57% vs 1.67%, P = 0.952). There were no significant differences in the rates of postoperative complications and mortality between the two groups. At median follow-up time of 60 months, stone recurrence was detected in one patient in group A and two in group B (1.79% vs 3.33%, P = 1.000). Stenosis of CBD was not observed in group A and slight stenosis in one patient in group B (0 vs 1.67%, P = 1.000)., Conclusion: Primary closure of the CBD upon completion of laparoscopic choledochotomy is safe and feasible in elderly patients ≥70 years old., (Copyright © 2019 First Affiliated Hospital, Zhejiang University School of Medicine in China. Published by Elsevier B.V. All rights reserved.)