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Abstract The analysis of genomic variations in offspring after implantation has been infrequently studied. In this study, we aim to investigate the extent of de novo mutations in humans from developing fetus to birth. Using high-depth whole-genome sequencing, 443 parent-offspring trios were studied to compare the results of de novo mutations (DNMs) between different groups. The focus was on fetuses and newborns, with DNA samples obtained from the families’ blood and the aspirated embryonic tissues subjected to deep sequencing. It was observed that the average number of total DNMs in the newborns group was 56.26 (54.17–58.35), which appeared to be lower than that the multifetal reduction group, which was 76.05 (69.70–82.40) (F = 2.42, P = 0.12). However, after adjusting for parental age and maternal pre-pregnancy body mass index (BMI), significant differences were found between the two groups. The analysis was further divided into single nucleotide variants (SNVs) and insertion/deletion of a small number of bases (indels), and it was discovered that the average number of de novo SNVs associated with the multifetal reduction group and the newborn group was 49.89 (45.59–54.20) and 51.09 (49.22–52.96), respectively. No significant differences were noted between the groups (F = 1.01, P = 0.32). However, a significant difference was observed for de novo indels, with a higher average number found in the multifetal reduction group compared to the newborn group (F = 194.17, P

Abstract Background Chromosomal microarray analysis (CMA) has emerged as a critical instrument in prenatal diagnostic procedures, notably in assessing congenital heart diseases (CHD). Nonetheless, current research focuses solely on CHD, overlooking the necessity for thorough comparative investigations encompassing fetuses with varied structural abnormalities or those without apparent structural anomalies. Objective This study sought to assess the relation of single nucleotide polymorphism-based chromosomal microarray analysis (SNP-based CMA) in identifying the underlying causes of fetal cardiac ultrasound abnormalities. Methods A total of 2092 pregnant women who underwent prenatal diagnosis from 2017 to 2022 were included in the study and divided into four groups based on the presence of ultrasound structural abnormalities and the specific type of abnormality. The results of the SNP-Array test conducted on amniotic fluid samples from these groups were analyzed. Results Findings from the study revealed that the non-isolated CHD group exhibited the highest incidence of aneuploidy, overall chromosomal abnormalities, and trisomy 18, demonstrating statistically significant differences from the other groups (p 0.05). The occurrence rates of 22q11.2 and 15q11.2 were also not statistically different between the isolated CHD group and the non-isolated congenital heart defect group (p > 0.05). Conclusion SNP-based CMA enhances the capacity to detect abnormal CNVs in CHD fetuses, offering valuable insights for diagnosing chromosomal etiology and facilitating genetic counseling. This research contributes to the broader understanding of the utility of SNP-based CMA in the context of fetal cardiac ultrasound abnormalities.

BackgroundCurrent research offers limited clarity on the correlation between waist circumference and chronic pain prevalence.ObjectiveThis investigation seeks to elucidate the potential relationship between waist circumference and chronic pain and their causal association.MethodsAn observational study was conducted, leveraging data from the National Health and Nutrition Examination Survey (NHANES) collected between 2001 and 2004. The multivariable logistic regression was used to assess the relationship between waist circumference and chronic pain. Furthermore, a meta-analysis of Mendelian Randomization (MR) was applied to explore a causal relationship between waist circumference and pain.ResultsThe observational study, post multivariable adjustment, indicated that an increase in waist circumference by 1 dm (decimeter) correlates with a 14% elevation in chronic pain risk (Odds Ratio [OR] = 1.14, 95% Confidence Interval [CI]: 1.04–1.24, p = 0.01). Moreover, the meta-analysis of MR demonstrated that an increased waist circumference was associated with a genetic predisposition to pain risk (OR = 1.14, 95%CI: 1.06–1.23, p = 0.0007).ConclusionObservational analysis confirmed a significant relationship between increased waist circumference and the incidence of chronic pain, and results based on MR Study identified increased waist circumference as potentially causal for pain.

Abstract Introduction Hypoxia‐inducible factor‐2α (HIF‐2α) modulates the hypoxic response pathway in tumors; however, mutations in pathways (including SDHA, SDHB, SDHC, SDHD, FH, and VHL genes) that are suspected to activate HIF‐2α are poorly understood, with limited understanding of the prevalence and clinical prognosis. Methods This retrospective observational study used a de‐identified nationwide (US‐based) clinico‐genomic database (CGDB) across 15 available tumor types. Results Among the 9467 adult patients with advanced/metastatic solid tumors included in the analysis, any mutation at the above‐mentioned six genes was observed in 1.8% (95% CI: 1.5–2.1) of patients. The mutation prevalence ranged from 0.05% of SDHD to 0.93% of VHL. When further stratified by tumor type, the prevalence of gene mutation in each tumor type was well below 1%, except for VHL with 44% in renal cell carcinomas (RCC). Excluding RCC, the prevalence of any HIF‐2α gene mutations in the study population was 0.9% (95% CI: 0.8–1.2). The median overall survival (OS) from 1 and 2 L therapy among patients with any HIF‐2α gene mutation was 14.5 (95% CI: 11.5–24.2) and 9.3 (95% CI: 6.0–18.1) months, respectively, compared with 13.4 (95% CI: 12.9–13.9) and 9.8 (95% CI: 9.3–10.4) months among patients without HIF‐2α gene mutations. Discussion and Conclusions The prevalence of HIF‐2α related gene mutations was generally low (

Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.

The effect of configuration entropy (ΔSconf) on thermal properties and magnetocaloric effects (MCE) of metallic glasses (MGs) remains vague. In this work, Tb55Co17.5Al27.5 low-, (Gd0.5Tb0.5)55Co17.5Al27.5 medium-, and (Gd1/3Tb1/3Dy1/3)55Co17.5Al27.5 high-entropy (LE, ME, and HE) MGs with critical diameters more than 5 mm were successfully fabricated. Systematic investigations into their thermodynamic behaviors suggest that high ΔSconf might deteriorate glass forming ability (GFA) along with reduced viscosity. Simultaneously, an improved thermal stability was observed in ME and HE MGs for their high activation energies for glass transition, crystal nucleation and growth, while the notable ordered structure in HE MG could accelerate glass transition and crystal growth by reducing activation energies. Studies of MCE indicate that the distinguished spin glass behaviors and higher structural order in HE MGs comprehensively contribute to an enhanced relative cooling power and a broader magnetic-transition temperature range, whereas the change of Curie temperature and maximum magnetic entropy more depends on the de Gennes factor rather than ΔSconf. All three designed MGs manifesting the competitive GFA and MCE could be candidates for magnetic refrigeration. Our studies show that entropy tailoring seems to play a dominant role in determining thermodynamic behaviors, which could help to design more stable MCE MGs.

Aims: To investigate the underling mechanisms of liver dysfunction in patients with polycystic ovary syndrome (PCOS).Materials and methods: PCOS patients were enrolled according to the Amsterdam criteria while PCOS animal model was established by dihydrotestosterone (DHEA) sustained release tablet implantation on its neck. Further liver damage and iron overload were detected by HE and Prussian blue staining. The liver related enzymes, mRNA and protein levels of hepcidin and GPX4 were tested by ELISA, qRT-PCR and Western blot. RNA interference and miR-761 transfection were routinely performed while the regulation of miR-761 on hepcidin and GPX4 was confirmed by luciferase reporter gene analysis.Results: We found that a part of PCOS patients and animal model had unexplained liver damage, which is independent of nonalcoholic fatty liver disease (NAFLD) and accompanied by increased ferrum (Fe) deposition. Besides, the expression of hepcidin and GPX4 that is important effector proteins for ferroptosis was down regulated in liver, showing the importance of iron metabolism in this unexplained liver damage. Based on the miR-761-hepcidin/GPX4 axis, we systematically studied the effects of miR-761 on ferroptosis and Fe deposition, which further influence the phenotype and liver function of PCOS model. From both in vivo and in vitro levels, changes in PCOS disease phenotype and ferroptosis were observed through hierarchical antagonism or overexpression of miR-761, hepcidin and GPX4.Conclusions: our results provide a novel explanation for unexplained liver damage in PCOS and a potential therapeutic target.

Abstract Background Decidual macrophages participate in immune regulation at the maternal–fetal interface. Abnormal M1/M2 polarization of decidual macrophages might predispose immune maladaptation in recurrent pregnancy loss (RPL). However, the mechanism of decidual macrophage polarization is unclear. We explored the role of Estradiol (E2)-sensitive serum-glucocorticoid regulated kinase (SGK) 1 in promoting macrophage polarization and suppressing inflammation at the maternal–fetal interface. Methods We assessed serum levels of E2 and progesterone during first trimester of pregnancy in women with or without threatened miscarriages (ended in live birth, n = 448; or early miscarriages, n = 68). For detection of SGK1 in decidual macrophages, we performed immunofluorescence labeling and western blot analysis applying decidual samples from RPL (n = 93) and early normal pregnancy (n = 66). Human monocytic THP-1 cells were differentiated into macrophages and treated with Toll-like receptor (TLR) 4 ligand lipopolysaccharide (LPS), E2, inhibitors or siRNA for in vitro analysis. Flow cytometry analysis were conducted to detect macrophages polarization. We also applied ovariectomized (OVX) mice with hormones exploring the mechanisms underlying the regulation of SGK1 activation by E2 in the decidual macrophages in vivo. Results SGK1 expression down regulation in the decidual macrophages of RPL was consistent with the lower concentration and slower increment of serum E2 from 4 to 12 weeks of gestation seen in these compromised pregnancies. LPS reduced SGK1 activities, but induced the pro-inflammatory M1 phenotype of THP-1 monocyte-derived macrophages and T helper (Th) 1 cytokines that favored pregnancy loss. E2 pretreatment promoted SGK1 activation in the decidual macrophages of OVX mice in vivo. E2 pretreatment amplified SGK1 activation in TLR4-stimulated THP-1 macrophages in vitro through the estrogen receptor beta (ERβ) and PI3K pathway. E2-sensitive activation of SGK1 increased M2 macrophages and Th2 immune responses, which were beneficial to successful pregnancy, by inducing ARG1 and IRF4 transcription, which are implicated in normal pregnancy. The experiments on OVX mice have shown that pharmacological inhibition of E2 promoted nuclear translocation of NF-κB in the decidual macrophages. Further more, pharmacological inhibition or knockdown of SGK1 in TLR4-stimulated THP-1 macrophages activated NF-κB by promoting its nuclear translocation, leading to increased secretion of pro-inflammatory cytokines involved in pregnancy loss. Conclusion Our findings highlighted the immunomodulatory roles of E2-activated SGK1 in Th2 immune responses by priming anti-inflammatory M2 macrophages at the maternal–fetal interface, resulting in a balanced immune microenvironment during pregnancy. Our results suggest new perspectives on future preventative strategies for RPL.

Spatial analysis of microbiomes at single cell resolution is challenging. Here the authors report a highly multiplexed method for spatial profiling, sequential error-robust fluorescence in situ hybridisation (SEER-FISH), and show that this allows mapping of microbial communities at micron-scale.

Maturation arrest (MA) is a subtype of non-obstructive azoospermia, and male infertility is a known risk factor for testicular tumors. However, the genetic basis for many affected individuals remains unknown. Here, we identified a deleterious hemizygous variant of X-linked retinoblastoma-binding protein 7 (RBBP7) as a potential key cause of MA, which was also found to be associated with the development of Leydig cell tumors. This mutation resulted in premature protein translation termination, affecting the sixth WD40 domain of the RBBP7 and the interaction of the mutated RBBP7 with histone H4. Decreased BRCA1 and increased γH2AX were observed in the proband. In mouse spermatogonial and pachytene spermatocyte-derived cells, deprivation of rbbp7 led to cell cycle arrest and apoptosis. In Drosophila, knockdown of RBBP7/Caf1-55 in germ cells resulted in complete absence of germ cells and reduced testis size, whereas knockdown of RBBP7/Caf1-55 in cyst cells resulted in hyperproliferative testicular cells. Interestingly, male infertility caused by Caf1-55 deficiency was rescued by ectopic expression of wild-type human RBBP7 but not mutant variants, suggesting the importance of RBBP7 in spermatogenesis. Our study provides insights into the mechanisms underlying the co-occurrence of MA and testicular tumors and may pave the way for innovative genetic diagnostics of these 2 diseases.

Abstract Background To investigate if the correlation between left and right cerebral tissue oxygen saturation (SctO2) was affected by one-lung ventilation (OLV) in patients undergoing lung cancer surgery. Methods Patients who underwent surgery for lung cancer were enrolled. Left and right SctO2 were collected during anesthesia. The primary outcome was the correlation between left and right SctO2 at 30 min after OLV which was analysed by Pearson correlation and linear regression model. Secondary outcomes included the trend of left–right SctO2 change over the first 30 min after OLV, correlation of left–right SctO2 during OLV for each patient; maximal difference between left–right SctO2 and its relationship with postoperative delirium. Results Left–right SctO2 was moderately correlated at baseline (r = 0.690, P

Manufactured sand, known for its artificial production, eco-friendliness, cost-effectiveness, and sustainability, serves as an excellent alternative to natural sand. Stone powder content plays a crucial role in determining the performance of manufactured sand, significantly enhancing concrete compaction and its ability to withstand environmental degradation. This study aims to explore the feasibility of using environmentally and economically advantageous manufactured sand in the construction of subway tunnel segments in coastal areas. It investigates the impact of stone powder content on the corrosion resistance of concrete made with manufactured sand under the combined influence of chloride salts and stray currents. The analysis covers corrosion tendencies, the post-rusting performance of reinforcement bars, the resistance of concrete to chloride transport, and microstructure properties, which are assessed through electron microscope scanning and mercury compression testing. The findings indicate that the corrosion resistance of manufactured sand concrete generally surpasses that of river sand concrete. Furthermore, stone powder content within the 3–8% range leads to a denser concrete microstructure, reduced porosity, lower free-chloride ion levels, increased polarization resistance of post-corrosion reinforcements, lower corrosion current density, and reduced mass loss of reinforcing bars. This research provides valuable theoretical support for promoting the use of environmentally friendly manufactured sand concrete in subway construction projects.

CLAVATA3/Embryo surrounding region (CLE) peptides, the largest family of plant peptide hormones, is critical in regulating cell differentiation and division in meristematic tissues in plants. In this study, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF MS) equipped with an electrospray ionization source (ESI) was used to investigate the fragmentation mechanism of five typical CLE peptides. The results showed that the strongest parent ion of CLE peptides was two or three fold protonated ion. Under the optimal collision energy (15-20 V), the collision-induced dissociation (CID) of CLE peptides mainly generate b-type and y-type fragment ions due to the cleavage of polypeptide bonds. Based on the MS/MS spectra of peptides standards, it was concluded that highly abundant product ions such as b3 and y4 ion were easily formed by the cleavage at the Xxx-Pro (Xxx denotes any other amino acid), Xxx-Hyp, Xxx-Gly or Gly-Xxx amide bond. In order to verify above fragment mechanism, a synthesized CLE-like peptide (CLEL) was tested and the mass spectrometry fragmentation behavior of experiment was completely in line with that of predictions. It could be seen that the fragmentation mechanism mentioned above had strong applicability to CLE family peptides. According to the structure activity relationship revealed by plant physiologist, the CLE residues R1/H1, P4, P7, P9, G6 and H12/N12 are critical for their function. Combined with the cleavage patterns of CLE peptides and the conserved amino acid residues mentioned above, y1, y4, b5, b6, y6 and y7 ion are valuable for qualitative analysis. So the mass spectrum fragmentation mode and fragment ions (y1, b3, y4, and y7 ion) of the CLE dodecapeptide motif in Arabidopsis were predicted, which would provide the necessary scientific basis for the quantitative and qualitative analysis of CLE peptides. Considering the highly conservative structure of the CLE family structure, a data-independent CLE peptides screening and identification strategy based on these mass spectrometry fragmentation behaviors was put forward. Herein, CLEL was used as an unknown peptide to initially verify the feasibility of above screening strategy. The deduced structure by MassLynx software was almost identical with real peptide sequence of CLEL but minor difference in order of the last four residues, which could be easily corrected based on the conserve structure of CLE peptide family. This work will be very helpful for biologists to search those hidden predicted CLE peptides in plant kingdom. Furthermore, the high-abundance product ions of CLE peptides, b3, y4 and b6 ion, are valuable for quantitative analysis.

BackgroundThe prediction model of postoperative pneumonia (POP) after lung cancer surgery is still scarce.MethodsRetrospective analysis of patients with lung cancer who underwent surgery at The Fourth Hospital of Hebei Medical University from September 2019 to March 2020 was performed. All patients were randomly divided into two groups, training cohort and validation cohort at the ratio of 7:3. The nomogram was formulated based on the results of multivariable logistic regression analysis and clinically important factors associated with POP. Concordance index (C-index), receiver operating characteristic (ROC) curve, calibration curve, Hosmer-Lemeshow goodness-of-fit test and decision curve analysis (DCA) were used to evaluate the predictive performance of the nomogram.ResultsA total of 1252 patients with lung cancer was enrolled, including 877 cases in the training cohort and 375 cases in the validation cohort. POP was found in 201 of 877 patients (22.9%) and 89 of 375 patients (23.7%) in the training and validation cohorts, respectively. The model consisted of six variables, including smoking, diabetes mellitus, history of preoperative chemotherapy, thoracotomy, ASA grade and surgery time. The C-index from AUC was 0.717 (95%CI:0.677-0.758) in the training cohort and 0.726 (95%CI:0.661-0.790) in the validation cohort. The calibration curves showed the model had good agreement. The result of DCA showed that the model had good clinical benefits.ConclusionThis proposed nomogram could predict the risk of POP in patients with lung cancer surgery in advance, which can help clinician make reasonable preventive and treatment measures.

ABSTRACT Transcriptional factors (TFs) and their regulons make up the gene regulatory networks. Here, we developed a method based on TF-directed activation-induced cytidine deaminase (AID) mutagenesis in combination with genome sequencing, called AIDmut-Seq, to detect TF targets on the genome. AIDmut-Seq involves only three simple steps, including the expression of the AID-TF fusion protein, whole-genome sequencing, and single nucleotide polymorphism (SNP) profiling, making it easy for junior and interdisciplinary researchers to use. Using AIDmut-Seq for the major quorum sensing regulator LasR in Pseudomonas aeruginosa, we confirmed that a few TF-guided C-T (or G-A) conversions occurred near their binding boxes on the genome, and a number of previously characterized and uncharacterized LasR-binding sites were detected. Further verification of AIDmut-Seq using various transcriptional regulators demonstrated its high efficiency for most transcriptional activators (FleQ, ErdR, GacA, ExsA). We confirmed the binding of LasR, FleQ, and ErdR to 100%, 50%, and 86% of their newly identified promoters by using in vitro protein-DNA binding assay. And real-time RT-PCR data validated the intracellular activity of these TFs to regulate the transcription of those newly found target promoters. However, AIDmut-Seq exhibited low efficiency for some small transcriptional repressors such as RsaL and AmrZ, with possible reasons involving fusion-induced TF dysfunction as well as low transcription rates of target promoters. Although there are false-positive and false-negative results in the AIDmut-Seq data, preliminary results have demonstrated the value of AIDmut-Seq to act as a complementary tool for existing methods. IMPORTANCE Protein-DNA interactions (PDI) play a central role in gene regulatory networks (GRNs). However, current techniques for studying genome-wide PDI usually involve complex experimental procedures, which prevent their broad use by scientific researchers. In this study, we provide a in vivo method called AIDmut-Seq. AIDmut-Seq involves only three simple steps that are easy to operate for researchers with basic skills in molecular biology. The efficiency of AIDmut-Seq was tested and confirmed using multiple transcription factors in Pseudomonas aeruginosa. Although there are still some defects regarding false-positive and false-negative results, AIDmut-Seq will be a good choice in the early stage of PDI study.

ABSTRACT The part of expression noise that is brought about by transcriptional regulation (represented here as NTR) is an important criterion for estimating the regulatory mode of a gene. However, characterization of NTR is an under-explored area, and there is little knowledge regarding the genome-wide NTR in the model pathogen Pseudomonas aeruginosa. Here, with a library of dual-color transcriptional reporters, we estimated the NTR for over 90% of the promoters in P. aeruginosa. Most promoters exhibit low NTR, while 42 and 115 promoters with high NTR were screened out in the exponential and the stationary growth phases, respectively. Specifically, a rearrangement of NTR was found in promoters involved in amino acid metabolism when bacteria enter the exponential phase. In addition, during the stationary phase, high NTR was found in a wide range of iron-related promoters involving siderophore synthesis and heme uptake, ExsA-regulated promoters involving bacterial virulence, and FleQ-regulated promoters involving biofilm development. We also found a large-scale negative dependence of transcriptional regulation between high-NTR promoters belonging to different functional categories. Our findings offer a global view of transcriptional heterogeneity in P. aeruginosa. IMPORTANCE The phenotypic diversity of Pseudomonas aeruginosa is frequently observed in research, suggesting that bacteria adopt strategies such as bet-hedging to survive ever-changing environments. Gene expression noise (GEN) is the major source of phenotypic diversity. Large GEN from transcriptional regulation (represented as NTR) represent an evolutionary necessity to maintain the copy number diversity of certain proteins in the population. Here, we provide a system-wide view of NTR in P. aeruginosa under nutrient-rich and stressed conditions. High NTR was found in genes involved in flagella biosynthesis and amino acid metabolism under both conditions. Specially, iron acquisition genes exhibited high NTR in the stressed condition, suggesting a great diversity of iron physiology in P. aeruginosa. We further revealed a global negative dependence of transcriptional regulation between those high-NTR genes under the stressed condition, suggesting a mutually exclusive relationship between different bacterial survival strategies.

Background: Alport syndrome (AS) is a hereditary renal basement membrane disease that can lead to end-stage renal disease in young adults. It can be diagnosed by genetic analysis, being mostly caused by mutations in COL4A3, COL­4A4, and COL4A5. To date, there is no radical cure for this disease. Objectives: The aim of this study was to avoid the transmission of AS within an affected family by selecting healthy embryos for uterine transfer. The embryos were identified by preimplantation genetic testing for monogenic disorders (PGT-M). Methods: We used next-generation sequencing (NGS) to identify mutations in the proband and his parents. The results of NGS were confirmed by Sanger sequencing. Targeted NGS combined with targeted single-nucleotide polymorphism haplotyping was used for the in vitro identification of COL4A5 mutations in human embryos to prevent their intergenerational transmission. Results: The c.349_359delGGACCTCAAGG and c.360_361insTGC mutations in COL4A5 were identified in a family affected by X-linked AS. Whole-genome sequencing by NGS with targeted haplotyping was performed on biopsied trophectoderm cells. A healthy baby was born after transfer of a single freeze-thawed blastocyst. Conclusions: The use of targeted NGS for identifying diagnostic markers combined with targeted haplotyping is an easy and efficient PGT-M method for preventing intergenerational transmission of AS.

IntroductionThe mortality rate of non-HIV-infected Pneumocystis jirovecii pneumonia (PCP) is high. This research aimed to develop and validate two clinical tools for predicting the risk of death and intensive care unit (ICU) admission in non-HIV-infected patients with PCP to reduce mortality.MethodsA retrospective study was conducted at Peking Union Medical College Hospital between 2012 and 2021. All proven and probable non-HIV-infected patients with PCP were included. The least absolute shrinkage and selection operator method and multivariable logistic regression analysis were used to select the high-risk prognostic parameters. In the validation, the receiver operating characteristic curve and concordance index were used to quantify the discrimination performance. Calibration curves were constructed to assess the predictive consistency compared with the actual observations. A likelihood ratio test was used to compare the tool and CURB-65 score.ResultsIn total, 508 patients were enrolled in the study. The tool for predicting death included eight factors: age, chronic lung disease, respiratory rate, blood urea nitrogen (BUN), lactate dehydrogenase (LDH), cytomegalovirus infection, shock, and invasive mechanical ventilation. The tool for predicting ICU admission composed of the following factors: respiratory rate, dyspnea, lung moist rales, LDH, BUN, C-reactive protein/albumin ratio, and pleural effusion. In external validation, the two clinical models performed well, showing good AUCs (0.915 and 0.880) and fit calibration plots. Compared with the CURB-65 score, our tool was more informative and had a higher predictive ability (AUC: 0.880 vs. 0.557) for predicting the risk of ICU admission.ConclusionIn conclusion, we developed and validated tools to predict death and ICU admission risks of non-HIV patients with PCP. Based on the information from the tools, clinicians can tailor appropriate therapy plans and use appropriate monitoring levels for high-risk patients, eventually reducing the mortality of those with PCP.

BackgroundBoth double-lumen tube (DLT) and bronchial blocker (BB) are used for lung isolation in patients undergoing lung cancer surgery. However, the effects of different devices for lung isolation remain inconclusive. Present study was designed to investigate the association between the choice of the two devices and postoperative pulmonary complications (PPCs) in patients with lung cancer.MethodsIn this retrospective cohort study, patients who underwent lung cancer surgery between January 1, 2020 and October 31, 2020 were screened. Patients were divided into two groups according to different devices for lung isolation: DLT group and BB group. Primary outcome was the incidence of a composite of PPCs during postoperative in-hospital stay.ResultsA total of 1721 were enrolled for analysis, of them, 868 received DLT and 853 BB. A composite of PPCs was less common in patients with BB (25.1%, [214/853]) than those received DLT (37.9% [329/868] OR 0.582 95% CI 0.461-0.735 P < 0.001). Respiratory infection was less common in BB group (14.4%, [123/853]) than DLT group (30.3%, [263/868], P

Emerging per- and polyfluoroalkyl substances (PFAS) alternatives are increasingly used in daily life. Although legacy PFAS have been associated with miscarriage in previous studies, it remains unknown whether exposure to emerging and legacy PFAS has any impact on the risk of unexplained recurrent spontaneous abortion (URSA). We conducted a case-control study with 464 URSA cases who had at least 2 unexplained miscarriages and 440 normal controls who had at least one normal livebirth. Concentrations of 21 PFAS in plasma, including three emerging PFAS alternatives, eight linear and branched PFAS isomers, four short-chain PFAS, and six legacy PFAS, were measured by ultra-performance liquid chromatography coupled with a tandem mass spectrometry (UPLC-MS/MS). Multiple logistic regression was applied to evaluate the relationship between PFAS and URSA risk. Perfluorooctanoic acid (PFOA, median: 6.18 ng/mL), perfluorooctane sulfonate (PFOS, median: 4.10 ng/mL), and 6:2 chlorinated perfluoroalkyl ether sulfonic acid (6:2 Cl-PFESA, median: 2.27 ng/mL) were the predominant PFAS in the controls. Exposure to 6:2 Cl-PFESA [adjusted odds ratio (aOR) = 1.18 (95% CI: 1.00, 1.39)] and hexafluoropropylene oxide dimer acid (HFPO-DA) [aOR = 1.35 (95% CI: 1.15, 1.59)] were significantly associated with increased risks of URSA. Women with older age (>30 years old) had a stronger association between PFAS and URSA. Our results suggest that emerging PFAS alternatives may be an important risk factor for URSA.

Abstract Objectives We analysed the peripheral blood lymphocyte subsets of human immunodeficiency virus (HIV)-negative patients infected with pneumocystis pneumonia (PCP) to determine the relationships between the levels of different types of lymphocytes and the prognosis of patients. Methods We retrospectively reviewed HIV-negative patients with PCP diagnosed in our department. All the eligible patients underwent lymphocyte subset analysis on admission. Results A total of 88 HIV-negative PCP patients were enrolled in the study. In univariate analyses, low CD4+ T cell count, low CD8+ T cell count, and low natural killer cell (NK cell) count were associated with higher in-hospital mortality. CD8+ T cell count ≤300/μL was found to be an independent risk factor for poor prognosis in multivariate logistical regression analysis (p = 0.015, OR = 11.526, 95% CI = 1.597–83.158). Although low CD4+ T cell and NK cell counts were not independent risk factors, the mortality rates of PCP patients decreased as the CD4+ T cell and NK cell counts increased. Conclusion The immune process of Pneumocystis jirovecii infection is complex but important. We propose that lymphocyte subsets could give clinicians a better understanding of patient immune status, helping with the early identification of potentially lethal infections and treatment decision making, such as adjusting the immunosuppressive regimen and choosing an appropriate patient monitoring level.

In laser tracking systems, capturing moving targets is a prerequisite to guaranteeing a tracking system’s performance. Previous studies have confirmed that the capture probability of hexagonal spiral scanning is higher than that for other scanning methods, but there is still room for improvement. This article proposes an improved hexagonal honeycomb scanning capture method based on hexagonal spiral scanning for a prior moving target model with a Gaussian distribution positioned within the scanning range of the visual threshold. Through experimental verification, it was found that, within the same scanning field of view, the capture probability can be increased by 3% compared to that in traditional hexagonal spiral scanning, making the capture probability greater than 98%. The improved hexagonal honeycomb structure scanning method proposed in this article provides a new solution for target acquisition problems in fields such as laser communication, laser docking, and airborne radar.

Free-form surfaces have good aberration correction capability and balance capability for nonrotationally symmetric imaging systems. In this study, we analyzed the quantitative relationship between X–Y polynomial combination and aberration for the efficient design of X–Y free-form optical systems. The purpose of this study was to provide an exhaustive design method for off-axis triple inverse optical systems with X–Y free-form surfaces. Finally, we designed a free-form off-axis optical system with a large field of view (FOV) and low distortion based on the off-axis triple inverse optical system without an intermediate image plane. The primary mirror (PM) of the system adopted an X–Y polynomial free-form surface to correct the aberration of different FOVs of the system and improve the image width and quality. The optical system had a focal length of 1000 mm, an F-value of 9.5, an FOV angle of 23° × 1°, a maximum distortion grid in the FOV less than or equal to −0.05%, and a full-field average wave aberration better than 0.055 λ (λ/18.2, λ = 632.8 nm). The analysis of the design results showed that the system had high-quality imaging and a compact structure. This design method can provide a technical reference for the study of such free-form off-axis systems.

Lithium−sulfur battery (LSB) is one of the most promising next‐generation batteries due to its high energy density and low cost. However, the serious shuttle effect, large volume change, and poor conductivity have hindered their commercialization. Herein, alk‐MXene@Fe3O4 composite is synthesized through ultrasonic treatment of alkalized MXene and Fe3O4 and it is used as additive in the sulfur cathode. The results indicate that Alk‐MXene@Fe3O4 additive can anchor polysulfides by the strong polar adsorption and accelerate the conversion of the LiPSs. The cathode with Alk‐MXene@Fe3O4 retains a substantially stable discharge capacity of 681.7 mAh g−1 after 120 cycles at a rate of 0.2 C, which is equivalent to 58.6% of the initial discharge capacity (1163 mAh g−1), and the corresponding capacity decay per cycle is 0.35%. At a rate of 1 C, it can still retain 595 mAh g−1 after 200 cycles. It has important reference value for the development of cathode additives to improve the performance of active sulfur.

Summary: Despite their widespread study, lithium metal anodes still face the bottleneck problem of low average Coulombic efficiency. Herein, we adopt an electroless plating method and introduce additive (vanillin) to develop nanoscale silver hosts. The uniform nanoscale silver layer is conducive to seamless Li/Ag alloying process, thereby enabling flat lithium deposition. Since the Li/Ag alloying process occurs after the formation of solid electrolyte interphase (SEI), the seamless alloying process stabilizes the SEI, which improves the reversibility of the lithium metal anode. On this basis, a 3D-printed copper host-coated dense nanoscale silver layer is constructed, which reduces the effective current density and hence lowers the polarization of lithium deposition or stripping. Moreover, the 3D structure induces the epitaxial growth of lithium and thus alleviates the volume change of lithium metal. As a result, an average Coulombic efficiency of 99.61% is achieved in the Li/3D Ag half-cell.

In order to explore the quantitative characterization method of non-oriented acrylic sheet fracture and the correlation between the fracture morphology of acrylic sheet and tensile temperature. The non-oriented acrylic sheet for aircraft was used as the research object. Firstly, the size and surface roughness of the mist region were measured by stereo microscope and 3D Laster Scanner(LEXT). Secondly, the fracture was measured by box counting method. Finally, the energy consumed by the rupture of the acrylic sheet to form the mist zone was estimated by two hypotheses and was related to the fractal dimension of the fractured mist zone. The results show that when the stretching rate is constant, the stretching temperature is raised from -55 ℃ to 60 ℃, the size of the fractured mist zone I3 is increased from 1.257 mm to 4.978 mm, and the surface roughness Raa is reduced from 0.517 μm to 0.330 μm, the fractal dimension D is increased from 1.357 to 1.579. The fitting degree factors of the fitting curves of the tensile temperature and the size, surface roughness and fractal dimension of the fracture foggy area are higher than 0.9, and there fitting degree of these curves are higher. Therefore, the morphological parameters of the tensile fracture of the acrylic sheet have a certain correlation with the fracture conditions, and the energy consumed in the formation of the mist region of the acrylic sheet is positively correlated with its fractal dimension.

Enhancing the circular dichroism signals of chiral plasmonic nanostructures is vital for realizing miniaturized functional chiroptical devices, such as ultrathin wave plates and high-performance chiral biosensors. Rationally assembling individual plasmonic metamolecules into coupled nanoclusters or periodic arrays provides an extra degree of freedom to effectively manipulate and leverage the intrinsic circular dichroism of the constituent structures. Here, we show that sophisticated manipulation over the geometric parameters of a plasmonic stereo-metamolecule array enables selective excitation of its surface lattice resonance mode either by left- or right-handed circularly polarized incidence through diffraction coupling, which can significantly amplify the differential absorption and hence the intrinsic circular dichroism. In particular, since the diffraction coupling requires no index-matching condition and its handedness can be switched by manipulating the refractive index of either the superstrate or the substrate, it is therefore possible to achieve dynamic tuning and active control of the intrinsic circular dichroism response without the need of modifying structure parameters. Our proposed system provides a versatile platform for ultrasensitive chiral plasmonics biosensing and light field manipulation.

Abstract Objective The aim of this study was to validate the results of two Emanuel syndromes detected by non-invasive prenatal screening (NIPS) screening using invasive methods, providing clinical performance of NIPS on chromosome microduplication detection. Methods NIPS was performed to diagnose the Emanuel syndrome. Amniocentesis or cordocentesis was performed to confirm the positive screening result of Emanuel syndrome cases. Fetal sample was detected by karyotyping, fluorescence in situ hybridization (FISH), and single nucleotide polymorphism array (SNP Array). Parental karyotyping and FISH were also carried out. Results Two cases with chromosomal abnormalities of 11q23.3q25 and 22q11.1q11.21 were found by NIPS. Chromosomal karyotyping showed that the two fetuses each have a small supernumerary marker chromosome (sSMC), SNP Array further demonstrated double duplications approximately 18 Mb in 11q23.3q25 and 3 Mb in 22q11.1q11.21. FISH confirmed that the small supernumerary marker chromosome (sSMC) was ish der(22)t(11;22) (TUPLE1+, ARSA-). Ultrasound scan and MRI showed some structure malformations in two fetuses. The two mothers were found to be a balanced carrier: 46,XX, t(11;22)(q23.3;q11.2). Conclusion NIPS could effectively identify Emanuel syndrome, which may indicate risks of a parent being a balanced rearrangement carrier. The followed confirmation test for positive sample is necessary and ensures the accuracy of the diagnosis.

IntroductionAssisted reproductive technology (ART) and previous Cesarean section (CS) are independently associated with the risk of adverse obstetric and perinatal outcomes in general. Few studies have focused on the association between adverse obstetric and perinatal outcomes and ART used in the high-risk population of women with previous CS.Materials and MethodsA retrospective cohort study including 14,099 women with a previous delivery and a subsequent delivery between April 2014 and April 2020 was conducted at our hospital. We assessed the risk of adverse obstetric and perinatal outcomes in pregnancies conceived by ART in women with previous CS, using log-binomial regression models.ResultsIn women with previous CS, ART singleton pregnancies were associated with an increased risk of maternal complications, such as pregnancy complications, placental anomalies of implantation, postpartum hemorrhage, and preterm birth (PTB), as compared to spontaneously conceived pregnancies. The implementation of ART and previous CS interacted in a synergistic manner to increase the likelihood of the placenta accreta spectrum in women with singleton pregnancies [adjusted relative risk (aRR) 5.30, 95% confidence interval (CI) 4.01–7.00; relative risk due to interaction: 1.41, 95%CI 0.07–2.75]. In women with previous CS who underwent ART, women with singletons conceived through intracytoplasmic sperm injection were at increased risk of velamentous placenta (aRR 2.46, 95%CI 1.35–4.48) compared with those with singletons conceived through in vitro fertilization (IVF), whereas women with singletons conceived through cleavage-stage embryo transfer (ET) were at increased risk of gestational diabetes mellitus (GDM) (aRR 1.74, 95%CI 1.16-2.60) than those with singletons conceived through blastocyst–stage ET.ConclusionPregnancies conceived through ART were at increased risk for adverse obstetric and perinatal outcomes in women who had previously delivered by CS, particularly for placental anomalies of implantation. In women with previous CS undergoing ART, IVF and blastocyst–stage ET may be a relatively safe treatment.

Abstract Objectives The effectiveness and costs of new treatments should be assessed in relation to existing practice. We describe treatments, survival and costs for advanced or metastatic small cell lung cancer (SCLC) patients receiving systemic therapy in the period preceding the introduction of immunotherapies. Materials and Methods This was a retrospective cohort study of patients aged ≥65 years, identified using linked Surveillance, Epidemiology, and End Results and Medicare databases. Individuals with a new primary diagnosis of SCLC between January 2007 and December 2013 were followed until December 2014. Chemotherapy treatments, health care visits and costs (in 2016 USD), and survival were determined by line of therapy. Results A total of 11 812 patients were identified with SCLC. First‐line (1L) chemotherapy was received by 6509 (55.1%) patients, most (93.2%) with carboplatin‐ (71.0%) or cisplatin‐ (22.2%) based therapies, typically combined with etoposide (79.2%). Second‐ (2L) and third‐ (3L) line chemotherapies were received by 2238 (18.9%) and 679 (5.7%) patients, of which 48.4% and 30.9%, respectively, were platinum‐based. The median durations of 1L, 2L, and 3L carboplatin‐based therapies were 5.9, 4.8, and 5.4 months, respectively, and the corresponding durations of cisplatin‐based therapies were 5.3, 4.2, and 5.3 months. During 1L, 2L, and 3L chemotherapies, patients averaged 8.2, 7.4, and 7.3 health care visits per month, respectively, and incurred total mean health care costs of $60 223, $42 636, and $35 903 per patient, respectively. Median survival from the start of 1L, 2L, and 3L chemotherapy was 9.2, 6.0, and 5.7 months, respectively. Conclusion First‐line chemotherapy was primarily platinum‐based, and a plethora of different regimens was used for 2L and 3L chemotherapies. Median survival from the start of 1L chemotherapy was 9 months, with an associated health care cost of $60 000. These data highlight an unmet medical need among SCLC patients receiving systemic therapy.

Objective This study aimed to compare the clinical outcomes for transfer of Day 3 (D3) double cleavage-stage embryos and Day 5/6 (D5/6) single blastocysts in the frozen embryo transfer (FET) cycle to formulate a more appropriate embryo transplantation strategy. Methods We retrospectively analyzed 609 FET cycles from 518 women from April 2017 to March 2021. All FETs were assigned to the D3-DET group (transfer of a Day 3 double cleavage-stage embryo), D5-SBT group (transfer of a Day 5 single blastocyst), or D6-SBT group (transfer of a Day 6 single blastocyst). Clinical outcomes were comparatively analyzed. Results There were no significant differences in the biochemical pregnancy rate, clinical pregnancy rate, or ongoing pregnancy rate between the D3-DET and D5-SBT groups, but these rates in the two groups were all significantly higher compared with those in the D6-SBT group. The implantation rate in the D5-SBT group was significantly higher than that in the D3-DET group. The twin pregnancy rate in the D5-SBT and D6-SBT groups was significantly lower than that in the D3-DET group. Conclusion This study suggests that D5-SBT is the preferred option for transplantation. D6-SBT reduces the pregnancy rate, making it a more cautious choice for transfer of such embryos.

Assisted reproductive technology (ART) has been linked to cholesterol metabolic and respiratory disorders later in life, but the mechanisms by which biosynthetic signaling remain unclear. Lung inflammatory diseases are tightly linked with the sterol regulatory element-binding protein (SREBP) and SREBP cleavage-activating protein (SCAP), but this has not been shown in an ART offspring. Here, mouse models from a young to old age were established including in vitro fertilization (IVF), intracytoplasmic injection (ICSI), and in vivo fertilized groups. In our results, significantly higher plasma levels of CRP, IgM, and IgG were identified in the aged ICSI mice. Additionally, pulmonary inflammation was found in four aged ART mice. At three weeks, ART mice showed significantly downregulated levels of Scap, Srebp-1a, Srebp-1c, and Srebf2 mRNA in the lung. At the same time, significant differences in the DNA methylation rates of Scap-Srebfs and protein expression of nuclear forms of SREBPs (nSREBPs) were detected in the ART groups. Only abnormalities in the expression levels of Srebp-1a and Srebp-1c mRNA and nSREBP1 protein were found in the ART groups at 10 weeks. However, at 1.5 years old, aberrant expression levels and DNA methylation of SCAP, SREBP1, and SREBP2, and their associated target genes, were observed in the lung of the ART groups. Our results indicate that ART increases long-term alterations in SCAP/SREBP expression that may be associated with their aberrant methylation status in mouse.

Objectives: The aim of this study was to investigate the efficacy of combination therapy of caspofungin and TMP/SMZ (trimethoprim/sulfamethoxazole) in moderate to severe pneumocystis jirovecii pneumonia (PJP) in patients without human immunodeficiency virus infection (HIV) and the relationship between therapeutic effect and plasma (1, 3) Beta-d-Glucan (BDG) levels. Methods: We retrospectively reviewed HIV-negative patients with PJP diagnosed in our department, who were treated with combination therapy of caspofungin and TMP/SMZ or monotherapy of TMP/SMZ during a six and a half year period. Results: A total of 126 moderate to severe PJP patients were enrolled in the study. In the multivariate analysis, low lymphocyte counts, high serum lactate dehydrogenase levels at the diagnosis of PJP and progression to shock were significant risk factors for death. In all patients, there was no significant difference in risk of death at 3 months. In the group of BDG ≥ 800 pg/m, patients receiving combination therapy was associated with a significantly decreased risk of death at 3 months, whereas in the group of BDG

Abstract Background Chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids (CLIPPERS) is an inflammatory disorder in the central nervous system (CNS) with distinct clinical, radiological, and pathological characteristics. The pathophysiology of CLIPPERS still remains unclear. Because a few cases about lymphoma mimicking the manifestations of CLIPPERS were reported and the prognosis of lymphoma is much worse, early identification of lymphoma is very important. Case presentation A 31-year-old woman was admitted with 3 months’ history of diplopia, dizziness, gait ataxia, and right facial numbness. The diagnosis of CLIPPERS was established based on the finding of punctate enhancing lesions in the cerebellum, thalamus, pons, medulla, and midbrain region in magnetic resonance imaging (MRI), together with the favorable clinical and radiological responses to corticosteroids. However, she was diagnosed as peripheral T cell lymphoma, not otherwise specified (PTCL-NOS) by the pulmonary nodular and the skin biopsy almost 10 years later, and she got complete remission within 1 year after chemotherapy. Conclusion We report the first case of CLIPPERS developing PTCL-NOS. This case proposes that when brain biopsy was difficult to achieve, biopsies in extra-cerebral lesions under the assisting examination of positron emission tomography-computed tomography (PET-CT) can be helpful in further identification.

Abstract Background Nail-patella syndrome (NPS) is an autosomal dominant developmental disorder most commonly characterized by dyplasia of nail or patella, the radial head or the humeral head hypoplasia, and, frequently ocular abnormalities and renal disease. It is caused by heterozygous loss-of-function mutations in the LMX1B gene, which encodes LIM homeodomain transcription factor and is essential for regulating the dorsal limb fate. Methods A five generation pedigree was recruited. Genomic DNA was extracted from the peripheral blood samples. Mutation detection was performed by Sanger sequencing the LMX1B gene. In silico functional annotation of the variant was performed using the in silico predictors SIFT, PolyPhen-2 and Mutation Taster. Results A novel heterozygous small deletion within exon 4 of LMX1B, c.712_714delTTC, was identified in a rare five-generation NPS pedigree. The mutation resulted in a deletion of the conserved amino acid phenylalanine at codon 238 (p.Phe238del), which located in the homeodomain of LMX1B may abolish DNA binding with the molecule. Conformational prediction showed that the variation could transform the helical structure comprising p.Phe234, p.Lys235, p.Ala236, and p.Ser237. Conclusion We identified a novel NPS-causing LMX1B mutation and expanded the spectrum of mutations in the LMX1B gene. The c.712_714delTTC mutation may affect the quaternary structure of LMX1B, which is essential for the specification of dorsal limb fate at both zeugopodal and autopodal levels, leading to typical NPS.

Amphihelical antimicrobial peptides (AMPs) are bactericidal host defense factors, but their function as immunomodulators is emerging. Here the authors show that several AMPs organize DNA into periodic nanocrystals by self-assembling into superhelical protofibril scaffolds, which potentiates DNA sensing by TLR9.

Image encryption is an efficient visual technology to protect private images. This paper develops an image encryption algorithm utilizing the principles of the Josephus problem and the filtering technology. The encryption algorithm follows the classical diffusion and confusion structure. The principle of Josephus problem is used to shuffle the image pixels to different positions to achieve the confusion property. Using a randomly generated filter, the filtering technology can spread slight changes of the original image to all pixels of the cipher image to obtain diffusion property. The simulation results show that the developed image encryption algorithm is able to encrypt different kinds of images into cipher images with uniform distribution. The security analysis demonstrates that it has an extremely sensitive secret key, can resist various security attacks, and has a better performance than several advanced image encryption algorithms.

Abstract Background Recent advances in semiconductor sequencing platform (SSP) have provided new methods for preimplantation genetic diagnosis/screening (PGD/S). The present study aimed to evaluate the applicability and efficiency of SSP in PGD/S. Methods The artificial positive single-cell-like DNAs and normal single-cell samples were chosen to test our semiconductor sequencing platform for preimplantation genetic diagnosis/screening (SSP-PGD/S) method with two widely used whole-genome amplification (WGA) kits. A total of 557 single blastomeres were collected from in vitro fertilization (IVF) couples, and their WGA products were processed and analyzed by our SSP-PGD/S method in comparison with array comparative genomic hybridization (array-CGH). Results Our SSP-PGD/S method indicated high compatibilities with two commercial WGA kits. For 557 single blastomeres, our method with four million reads in average could detect 24-chromosome aneuploidies as well as microdeletion/microduplication of the size over 4 Mb, providing 100% consistent conclusion with array-CGH method in the classification of whether it was transplantable. Conclusions Our studies suggested that SSP-PGD/S represents a valuable alternative to array-CGH and brought PGD/S into a new era of more rapid, accurate, and economic.

Phosphorylation of intrinsically disordered proteins (IDPs) can produce changes in structural and dynamical properties and thereby mediate critical biological functions. How phosphorylation effects intrinsically disordered proteins has been studied for an increasing number of IDPs, but a systematic understanding is still lacking. Here, we compare the collapse propensity of four disordered proteins, Ash1, the C-terminal domain of RNA polymerase (CTD2'), the cytosolic domain of E-Cadherin, and a fragment of the p130Cas, in unphosphorylated and phosphorylated forms using extensive all-atom molecular dynamics (MD) simulations. We find all proteins to show V-shape changes in their collapse propensity upon multi-site phosphorylation according to their initial net charge: phosphorylation expands neutral or overall negatively charged IDPs and shrinks positively charged IDPs. However, force fields including those tailored towards and commonly used for IDPs overestimate these changes. We find quantitative agreement of MD results with SAXS and NMR data for Ash1 and CTD2' only when attenuating protein electrostatic interactions by using a higher salt concentration (e.g. 350 mM), highlighting the overstabilization of salt bridges in current force fields. We show that phosphorylation of IDPs also has a strong impact on the solvation of the protein, a factor that in addition to the actual collapse or expansion of the IDP should be considered when analyzing SAXS data. Compared to the overall mild change in global IDP dimension, the exposure of active sites can change significantly upon phosphorylation, underlining the large susceptibility of IDP ensembles to regulation through post-translational modifications.

Nowadays, most of the preimplantation genetic testing (PGT) is performed with a strategy of comprehensive chromosome screening and trophectoderm biopsy. Nevertheless, patients with ovarian insufficiency may not have competent blastocysts. In the present study, we aimed to establish the value of multiple annealing and looping-based amplification cycle (MALBAC)-based next-generation sequencing (NGS) for PGT in day-3 embryos. A total of 94.3% (1168/1239) of embryos yielded informative results, and the overall embryo euploid rate was 21.9% (256/1168). Overall, 225 embryos were transferred in 169 cycles with a clinical pregnancy rate of 49.1% (83/169). The live birth and implantation rates were 47.3% (80/169) and 44.4% (100/225), respectively. Double embryos transfer showed higher clinical pregnancy and live birth rates compared with single embryo transfer, but the implantation rates were similar (44.2% vs. 44.6%, P > 0.05). The euploid rate for reciprocal translocations (16.1%) was significantly lower than that for Robertsonian translocations (28.0%, P < 0.01) and inversions (28.0%, P < 0.01). However, higher percentages of embryos with de novo abnormalities were observed with Robertsonian translocations (23.3%, P < 0.01) and inversions (30.5%, P < 0.01) than with reciprocal translocations (11.6%). We demonstrated that NGS for PGT on day-3 embryos is an effective clinical application, particularly for patients with a diminished ovarian reserve and limited embryos.

Aim: PD-L1 expression and high levels of microsatellite instability (MSI-H) may predict response to checkpoint inhibitors, but their prevalence and prognostic value are unknown in many cancers. Methods: We retrospectively evaluated PD-L1 combined positive score (CPS) and MSI-H and their association with clinical outcomes among patients with ten advanced uncommon cancers. Results: 398 of 426 patients (93%) had a valid PD-L1 result; most (242; 61%) had CPS ≥1. Prevalence of MSI-H tumors was 8/360. Median overall survival was shorter among patients with PD-L1 CPS ≥1 tumors after first-line treatment (23.0 vs 39.7 months, p = 0.014). Conclusion: PD-L1 was commonly expressed in solid tumors, and CPS ≥1 was associated with shorter overall survival. Prevalence of MSI-H was low.

The impact of aging on reproductive outcomes has received considerable critical attention; however, there is much less information available on the effects of paternal age compared to the effects of maternal age. In this study, methylation levels of sperm rDNA promoter regions and Long Interspersed Nucleotide Element 1 (LINE-1) were measured using pyrosequencing and fertilization, day 3 good-quality embryo, pregnancies, and implantation results were assessed. We observed significantly increasing levels of DNA methylation in the sperm rDNA promoter regions with age based on stratifying the samples by age alone (P = 0.0001) and performing linear regression analysis (P < 0.0001). Meanwhile, no statistically significant correlations were observed between global LINE-1 methylation with age. No statistically significant correlations were observed between sperm rDNA promoter methylation levels and either the day 3 good-quality embryo rate or clinical pregnancy rate. In contrast, the correlation between sperm rDNA promoter methylation levels and fertilization (2 pronuclei) rate was nearly significant (P = 0.0707), especially the methylation levels of some individual CpG units (CpG_10, P = 0.0176; CpG_11, P = 0.0438; CpG_14, P = 0.0232) and rDNA promoter methylation levels measured using primerS2 (P = 0.0513). No significant correlation was found between sperm rDNA promoter methylation levels and fertilization rates (2 pronuclei, 1 pronuclei, and 1 polypronuclei). Our results demonstrate that sperm are susceptible to age-associated alterations in methylation levels of rDNA promoter regions, suggesting that sperm rDNA promoter methylation levels can be applied to DNA methylation-based age prediction, and that the aberrant methylation of rDNA promoters may be partially responsible for enhanced disease susceptibility of offspring sired by older fathers. Methylation levels of sperm rDNA promoter regions may correlate with polypronuclei rates of IVF programs.