As the development of molecular diagnostic methods, a large number of clinically relevant or disease-related copy number variations (CNVs) could be detected, and the demand for genetic counselling and clinical treatment is also increasing. For patients with pathogenic or likely pathogenic CNVs, preimplantation genetic testing (PGT) could provide a feasible path to prevent the inheritance of the genetic disorder in the offspring. In this study, we included a couple with 1q21.1 recurrent microduplication to conduct molecular diagnosis and PGT clinical application. The optical genome mapping (OGM) successfully verified the orientation and location of the microduplication, which further proved OGM as a promising approach for chromosomal anomalies detection with high resolutions. In PGT application, linkage-analysis-based PGT and high resolution PGT-A were simultaneously conducted for the pedigree and all the embryos. The results were consistent between linkage analysis and high resolution aneuploid analysis in the targeted region. One embryo that was absent of paternal 1q21.1q21.2 duplication was selected for further transplantation. This successful clinical practice in this study shed light for future molecular diagnosis and PGT application in tandem microduplications.

As one of symbolic design features of Generation-III and III+ (G-III and G-III + ) nuclear reactors, the passive safety system has been implemented on majority of G-III and III + PWRs and BWRs. The passive safety systems in those reactors are designed differently yet adhere to identical fundamental principles. It would be interesting to examine the similarities and differences between the passive safety systems of G-III + PWR and BWR. System code simulations for Small Break and Large Break Loss of Coolant Accident for the Economic Simplified Boiling Water Reactor (ESBWR) and China's advanced PWR had been conducted, respectively. The results showed that the passive safety systems of ESBWR and China's Advanced PWR were comparable in design, and when dealing with the similar type of SBLOCA and LBLOCA, the passive safety components with the same functionality could all contribute to the core's cooling with stability and reliability, and the injection flow rate values were nearly equivalent. Separating the injection lines of ESBWR's safety systems guarantees that the action sequence could well remain stable. This design could prevent the injection flow rate and action sequence from being impacted by the interaction of safety injection systems. This work could serve as a reference for the design of subsequent new reactors, thereby contributing to the design of reactors with enhanced economics and safety.

Digital Twin (DT) technology has gained significant attention in various areas, including nuclear energy. Technologies such as machine learning, artificial intelligence, and high-fidelity modeling and simulation have been studied to further the digitalization of the nuclear industry. In this paper, an overview of the DT initiatives and developments for the design optimization and intelligent transformation of nuclear energy systems is described to provide a development trend of DT technology for nuclear reactors. For the design of DT for complex systems such as nuclear reactors, Model Based Systems Engineering (MBSE) technology offers requirements models, design processes, and system verification that are clearer, more precise, and more comprehensive. Four properties of Digital Twin for Nuclear Reactor System (DTNRS) are summarized, which are given by the block definition diagram of MBSE philosophy. Then, to bridge the gap between numerical models and theoretical research of DT, a systematic and hierarchical “V” architecture is proposed, which reflects the digital-related technologies of the DTNRS throughout the whole life cycle. Furthermore, a four-layered decomposition construction model is developed and a prospective application scenario of DTNRS is pictured. Finally, the five-step instructive demonstration of the DTNRS is given based on a Generation III (GEN-III) reactor with its visualized condition map.

ABSTRACT Objective Currently, the most commonly used methods for linkage analysis of pre‐implantation genetic testing for monogenic disorders (PGT‐M) are next generation sequencing (NGS) and SNP array. We aim to investigate whether the application efficacy of Asian screening array (ASA) in PGT‐M preclinical workup for the Chinese population is superior to NGS based single nucleotide polymorphism (SNP) panels. Methods We conducted a retrospective analysis by reviewing 294 couples from a single center over the past 4 years and compared the detection results between NGS‐based SNP panels and ASA. Using the numbers of informative SNPs upstream and downstream flanking of variants, we assessed the detection efficiency of both methods in monogenic diseases, chromosomal microdeletion syndrome and males with de novo variants, among other scenarios. Results Results indicate that ASA offers a greater number of informative SNPs compared with NGS‐based SNP panels. Additionally, data analysis for ASA is generally more straightforward and may require less computational resources. While ASA can address most PGT‐M challenges, we have also identified certain genes in previous tests that are not suitable for PGT‐M using ASA. Conclusion The application of ASA in PGT‐M preclinical workup for Chinese populations has good practical value as it can perform linkage analysis for most genetic variants. However, for certain variants, NGS or other testing methods, such as mutated allele revealed by sequencing with aneuploidy and linkage analysis (MARSALA), may still be necessary for completion.

Background Couples with balanced chromosome rearrangement (BCR) are at high risk of recurrent miscarriages or birth defects due to chromosomally abnormal embryos. This study aimed to provide real-world evidence of the euploidy rate of blastocysts from couples with BCR using preimplantation genetic testing (PGT) and to guide pretesting genetic counselling. Methods A continuous four-year PGT data from couples with BCR were retrospectively analyzed. Biopsied trophectoderm cells were amplified using whole genome amplification, and next-generation sequencing was performed to detect the chromosomal numerical and segmental aberrations. Clinical data and molecular genetic testing results were analyzed and compared among the subgroups. Results A total of 1571 PGT cycles with 5942 blastocysts were performed chromosomal numerical and segmental aberrations detection during the four years. Of them, 1034 PGT cycles with 4129 blastocysts for BCR couples were included; 68.96% (713/1034) PGT cycles had transferable euploid embryos. The total euploidy rate of blastocysts in couples carrying the BCR was 35.29% (1457/4129). Couples with complex BCR had euploid blastocyst rates similar to those of couples with non-complex BCR (46.15% vs. 35.18%, P > 0.05). Chromosome inversion had the highest chance of obtaining a euploid blastocyst (57.27%), followed by Robertsonian translocation (RobT) (46.06%), and the lowest in reciprocal translocation (RecT) (30.11%) ( P < 0.05). Couples with males carrying RobT had higher rates of euploid embryo both in each PGT cycles and total blastocysts than female RobT carriers did, despite the female age in male RobT is significant older than those with female RobT ( P < 0.05). The proportions of non-carrier embryos were 52.78% (95/180) and 47.06% (40/85) in euploid blastocysts from couples with RecT and RobT, respectively ( P > 0.05). RecT had the highest proportion of blastocysts with translocated chromosome-associated abnormalities (74.23%, 1527/2057), followed by RobT (54.60%, 273/500) and inversion (30.85%, 29/94) ( P < 0.05). Conclusions In couples carrying BCR, the total euploidy rate of blastocysts was 35.29%, with the highest in inversion, followed by RobT and RecT. Even in couples carrying complex BCR, the probability of having a transferable blastocyst was 46.15%. Among the euploid blastocysts, the non-carrier ratios in RecT and RobT were 52.78% and 47.06%, respectively. RecT had the highest proportion of blastocysts with translocated chromosome-associated abnormalities.

Introduction: X-linked recessive chronic granulomatous disease (XR-CGD) is a severe primary immunodeficiency principally caused by a CYBB (OMIM: 300481) gene variant. Recurrent fatal bacterial or fungal infections are the main clinical manifestations of XR-CGD. Patient concerns: In the current case, in vitro fertilization (IVF) associated with preimplantation genetic testing for monogenic disorder (PGT-M) was applied for a Chinese couple who had given birth to a boy with XR-CGD. Diagnosis: Next-generation sequencing-based SNP haplotyping and Sanger-sequencing were used to detect the CYBB gene variant (c.804 + 2T>C, splicing) in this family. Interventions: The patient was treated with IVF and PGT-M successively. Outcomes: In this IVF cycle, 7 embryos were obtained, and 2 of them were euploid and lacked the CYBB gene variant (c.804 + 2T>C). The PGT results were verified by prenatal diagnosis after successful pregnancy, and a healthy girl was eventually born. Conclusion: PGT-M is an effective method for helping families with these fatal and rare inherited diseases to have healthy offspring. It can availably block the transmission of disease-causing loci to descendant.

Best Estimate Plus Uncertainty analysis has been used in the design, certification, and safety review of Nuclear Power Plants. To analyze the small break loss of coolant accident of a Generation III + Boiling Water Reactor, a streamlined BEPU method has been established and partially demonstrated to a Bottom Drain Line small break Loss Of Coolant Accident post-test analysis in this investigation. A RELAP5 input model was built for an integral test facility simulating a Generation III + Reactor with passive safety features. The Fast Fourier Transform Based Method was utilized to preliminarily evaluate the accuracy of the code simulation results. The DAKOTA toolkit was utilized to drive uncertainty quantification and sensitivity analysis. The simulation results generally reproduce the experimental phenomenon observed during experiments. The uncertainty bands of output parameters were established to demonstrate the enclosing of experimental data and the safety margin according to the design limits.

Chromosome rearrangement is one of the main causes of abortion. In individuals with double chromosomal rearrangements, the abortion rate and the risk of producing abnormal chromosomal embryos are increased. In our study, preimplantation genetic testing for structural rearrangement (PGT-SR) was performed for a couple because of recurrent abortion and the karyotype of the male was 45, XY der (14; 15)(q10; q10). The PGT-SR result of the embryo in this in vitro fertilization (IVF) cycle showed microduplication and microdeletion at the terminals of chromosomes 3 and 11, respectively. Therefore, we speculated whether the couple might have a cryptic reciprocal translocation which was not detected by karyotyping. Then, optical genome mapping (OGM) was performed for this couple, and cryptic balanced chromosomal rearrangements were detected in the male. The OGM data were consistent with our hypothesis according to previous PGT results. Subsequently, this result was verified by fluorescence in situ hybridization (FISH) in metaphase. In conclusion, the male’s karyotype was 45, XY, t(3; 11)(q28; p15.4), der(14; 15)(q10; q10). Compared with traditional karyotyping, chromosomal microarray, CNV-seq and FISH, OGM has significant advantages in detecting cryptic and balanced chromosomal rearrangements.

The Pressurizer Surge Line Break (PSLB) Loss of Coolant Accident (LOCA) is one of the representative design basis accidents (DBAs) which leads to the discharge of the primary coolant into the containment atmosphere. In this paper, the RELAP5 best-estimate code was used to perform a PSLB LOCA on China's Advanced Pressurized Water Reactor (PWR). According to the phenomenon identification and ranking table (PIRT), the parameters that contribute significantly to the reactor safety standard were determined. Then, simple random sampling was performed using DAKOTA software and a set of 59 calculations were performed based on the Wilks' formula. The uncertainty bands of key output parameters were obtained with 95/95 criterion, and the upper band of maximum containment pressure did not exceed the safety limit. Lastly, important uncertainty contributors for the maximum containment pressure were identified through sensitivity analysis.

The autosomal dominant form of polycystic kidney disease (ADPKD) is the most common hereditary disease that causes late-onset renal cyst development and end-stage renal disease. Preimplantation genetic testing for monogenic disease (PGT-M) has emerged as an effective strategy to prevent pathogenic mutation transmission rely on SNP linkage analysis between pedigree members. Yet, it remains challenging to establish reliable PGT-M methods for ADPKD cases or other monogenic diseases with de novo mutations or without a family history. Here we reported the application of long-read sequencing for direct haplotyping in a female patient with de novo PKD1 c.11,526 G > C mutation and successfully established the high-risk haplotype. Together with targeted short-read sequencing of SNPs for the couple and embryos, the carrier status for embryos was identified. A healthy baby was born without the PKD1 pathogenic mutation. Our PGT-M strategy based on long-read sequencing for direct haplotyping combined with targeted SNP haplotype can be widely applied to other monogenic disease carriers with de novo mutation.

Melanoma is a highly malignant neoplasm of melanocytes with considerable metastatic potential and drug resistance, explaining the need for new candidates that inhibit tumor growth and metastasis. The signal transducer and activator of the transcription 3 (Stat3) signaling pathway plays an important role in melanoma and has been validated as promising anticancer target for melanoma therapy. In this study, nifuroxazide, an antidiarrheal agent identified as an inhibitor of Stat3, was evaluated for its anti-melanoma activity in vitro and in vivo. It had potent anti-proliferative activity against various melanoma cell lines and could induce G2/M phase arrest and cell apoptosis. Moreover, nifuroxazide markedly impaired melanoma cell migration and invasion by down-regulating phosphorylated-Src, phosphorylated-FAK and expression of matrix metalloproteinase (MMP) -2, MMP-9 and vimentin. It also significantly inhibited tumor growth without obvious side effects in the A375-bearing mice model by inducing apoptosis and reducing cell proliferation and metastasis. Notably, nifuroxazide significantly inhibited pulmonary metastases, which might be associated with the decrease of myeloid-derived suppressor cells (MDSCs). These findings suggested that nifuroxazide might be a potential agent for inhibiting the growth and metastasis of melanoma.

Preimplantation genetic testing (PGT) has been increasingly used to prevent rare inherited diseases. In this study, we report a case where PGT was used to prevent the transmission of disease-caused variant in a SCID-X1 (OMIM:300400) family. SCID-X1 is an X-linked recessive inherited disease whose major clinical manifestation of immune deficiency is the significant reduction in the number of T-cells and natural killer cells. This family gave birth to a boy who was a hemizygous proband whose IL2RG gene was mutated (c.315T > A, p(Tyr105*), NM_000206.3, CM962677). In this case, Sanger sequencing for mutated allele and linkage analysis based on single-nucleotide polymorphism (SNP) haplotype via next-generation sequencing were performed simultaneously. After PGT for monogenic disorder, we detected the aneuploidy and copy number variation (CNV) for normal and female carrier embryos. Four embryos (E02, E09, E10, and E11) were confirmed without CNVs and inherited variants at the IL2RG gene. Embryo E02 (ranking 4BB) has been transferred after considering the embryo growth rate, morphology, and PGT results. Prenatal genetic diagnosis was used to detect amniotic fluid cells, showing that this fetus did not carry the variant of the IL2RG gene (c.315T > A). Ultimately, a healthy girl who had not carried disease-causing variants of SCID-X1 confirmed by prenatal diagnosis was born, further verifying our successful application of PGT in preventing mutated allele transmission for this SCID family.

Thalassemia is a group of common hereditary anemias that cause significant morbidity and mortality worldwide. However, precisely diagnosing thalassemia, especially rare thalassemia variants, is still challenging. Long-range PCR and long-molecule sequencing on the PacBio Sequel II platform utilized in this study could cover the entire HBA1 , HBA2 and HBB genes, enabling the diagnosis of most of the common and rare types of thalassemia variants. In this study, 100 cases of suspected thalassemia were subjected to traditional thalassemia testing and third-generation sequencing for thalassemia genetic diagnosis. Compared with traditional diagnostic methods, an additional 10 cases of rare clinically significant variants, including 3 cases of structure variants and 7 cases of single nucleotide variations (SNVs) were identified, of which a case with − α 3.7 subtype III (− α 3.7III ) was first identified and validated in the Chinese population. Other rare variants of 11.1 kb deletions (− 11.1/αα), triplicate α-globin genes (aaa 3.7 /αα) and rare SNVs have also been thoroughly detected. The results showed that rare thalassemia variants are not rare but have been misdiagnosed by conventional methods. The results further validated third-generation sequencing as a promising method for rare thalassemia genetic testing.

Abstract Thalassemia is a group of common hereditary anemia that cause significant morbidity and mortality around the world, yet precisely diagnosing of thalassemia especially for rare thalassemia variants are still challenging. Long range PCR and long-molecule sequencing on the PacBio Sequel II platform utilized in this study could cover the entire HBA1, HBA2 and HBB gene which enable to diagnose most of common and rare types of thalassemia variants. In this study, 100 cases of suspected thalassemia were conducted traditional thalassemia testing and third generation sequencing for thalassemia genetic diagnosing. Compared with traditional diagnostic methods, here additional 10 cases of rare clinically significant variants, including 3 cases of structure variants (SVs) and 7 cases of single nucleotide variations (SNVs) were identified, of which a case with -α3.7 subtype III (-α3.7III) was first identified and validated in Chinese population. Other rare variants of 11.1 kb deletions (--11.1/αα), triplicate α-globin genes (aaa3.7/αα) and rare SNVs have also been thoroughly detected. The results showed that rare thalassemia variants are not rare but just been miss-diagnosed by conventional methods. The results further validated third generation sequencing as a promising method for rare thalassemia genetic testing.

Targeting phosphatidylinositol 3-kinase δ (PI3Kδ) is an important therapeutic strategy for indolent non-Hodgkin lymphomas (NHLs). However, we previously observed reactivation of phosphatidylinositol 3-kinase (PI3K) pathways in aggressive NHL cell lines following continuous exposure to PI3Kδ inhibitors (PI3Kδi), which limited their efficacy and suggests that more studies should be focused on this reactivation to improve current PI3Kδi-based treatments. Herein we conducted a drug synergy screening that combined a marketed PI3Kδi, idelalisib, with 14 well-characterized epigenetic drugs across several types of aggressive NHL cell lines. We identified BRD4 inhibitors (BRD4i) as potent partners that, in combination with idelalisib, were capable of synergistically exerting anti-proliferative activity and inducing cell apoptosis in a panel of aggressive NHL cell lines through continuous suppression of PI3K pathways. More importantly, the combination of BRD4i and PI3Kδi simultaneously inhibited transcription and translation of the oncogenic transcription factor c-MYC, downregulating the expression of c-MYC and continuously suppressing the proliferation of cancer cells in vitro, as well as the growth of tumors in vivo even after drug withdrawal. This study, thus, reveals the potential of simultaneously targeting PI3Kδ and BRD4 as a new therapeutic strategy for aggressive forms of NHL.

Diffuse large B-cell lymphoma (DLBCL) is a clinically aggressive B-cell non-Hodgkin's lymphoma (NHL) with high treatment difficulty and high relapse rate. The bromodomain and extra-terminal (BET) proteins play significant roles in supporting the transcription of known DLBCL oncogene MYC, which provides a way for the development of targeted therapeutic agents to address this kind of malignant tumor. Here, we reported a novel benzoxazinone derivative YLT-LL-11 as potential BRD4 inhibitor and further investigated the biological activities against DLBCL. The results suggested that YLT-LL-11 inhibited cell growth against a panel of human hematopoietic malignancies cell lines in a dose- and time-dependent manner. In addition, flow cytometry and Western blotting assays showed that YLT-LL-11 inhibited the proliferation of a DLBCL cell line OCI-LY10 via inducing G0/G1 cell cycle arrest with regulation of the cyclin-dependent kinases (CDKs) expression. Furthermore, YLT-LL-11 facilitated OCI-LY10 cell apoptosis by up-regulation of pro-apoptotic protein BAX and down-regulation of anti-apoptotic protein Bcl-2. Taken together, these results revealed that BRD4 inhibitor YLT-LL-11 can down-regulate growth-associated transcription factors MYC in DLBCL thus resulted in cell growth inhibition and apoptosis.

Background: Targeting phosphatidylinositol 3-kinase δ (PI3Kδ) is an important therapeutic strategy for indolent non-Hodgkin lymphomas (iNHL). However, reactivation of the PI3K pathway in aggressive NHL cell lines has been observed after continuous exposure to PI3Kδ inhibitors (PI3Kδi), which limits their efficacy. Hence, more studies focused on this problem are required to improve current PI3Kδi-based treatment.Methods: We conducted a drug synergy screening combining a marketed PI3Kδi, idelalisib, with 14 well-characterised epigenetic drugs using several types of aggressive NHL cell lines.Findings: We identified bromodomain-containing 4 (BRD4) inhibitors (BRD4i) as potent adjuvants, which exerted synergistic anti-tumour activity in combination with idelalisib and induced apoptosis in a panel of aggressive NHL cell lines by continuously suppressing the PI3K pathway. More importantly, co-treatment with BRD4i and PI3Kδi simultaneously inhibited the transcription and translation of the oncogenic transcription factor MYC proto-oncogene, bHLH transcription factor (c-MYC), to synergistically downregulate its expression. Furthermore, this action continuously suppressed cancer cell proliferation in vitro and tumour growth in vivo even after drug withdrawal.Interpretation: Thus, the findings of this study reveal the potential efficacy of simultaneously targeting PI3Kδ and BRD4 as a therapeutic strategy for aggressive forms of NHL.Funding Statement: This work was supported by Fundamental Research Funds for the Central Universities (No. A0920502051903-37). This work was also supported by the China Postdoctoral Science Foundation (grant number 2019M652543) and Sichuan Provincial Science and technology program for Key research and development, China (No. 2018SZ0007).Declaration of Interests: The authors declare that they have no potential conflicts of interest.Ethics Approval Statement: Mice maintenance were performed in a specific-pathogen-free (SPF) condition facility and all animal experiments have been approved by the Institutional Animal Care and Treatment Committee of Sichuan University in China (Permit Number: 20170135), which were carried out in accordance with the approved guidelines.

Triple negative breast cancer (TNBC) patients have a high risk of brain metastases. This deadly disease represents a major challenge for successful treatment, in part because of the poor ability of drugs to penetrate the blood-brain barrier. Antipsychotic drugs show good bioavailability in the brain, and some of them have exhibited anticancer effects in several cancer types. In this study, we investigated the potential of repurposing fluphenazine hydrochloride (Flu) for the treatment of TNBC and the brain metastases. Our data showed that Flu inhibited survival of metastatic TNBC cells. It induced G0/G1 cell cycle arrest and promoted mitochondria-mediated intrinsic apoptosis in vitro. Pharmacokinetic studies in BALB/c mice showed a brain/plasma drug concentration ratio of Flu above 25 for at least 24 hours after dosing. Flu moderately suppressed tumor growth in a TNBC subcutaneous xenograft mouse model. Importantly, Flu exhibited good anti-metastatic potential in a mouse brain metastasis model with an inhibition rate of 85%. In addition, Flu showed a strong inhibitory effect on spontaneous lung metastasis. Moreover, Flu didn't cause serious side effects in the mice. Taken together, this study prompts further preclinical and clinical investigation into repurposing Flu for treating metastatic TNBC patients, which urgently need new treatment options.

New analogues of antitubercular drug Delamanid were prepared, seeking drug candidates with enhanced aqueous solubility and high efficacy. The strategy involved replacement of phenoxy linker proximal to the 2-nitroimidazooxazole of Delamanid by piperidine fused 5 or 6-membered ring heterocycles (ring A). The new compounds were all more hydrophilic than Delamanid, and several class of analogues showed remarkable activities against M. bovis. And among these series, the tetrahydro-naphthyridine-linked nitroimidazoles displayed excellent antimycobacterial activity against both replicating (MABA) and nonreplicating (LORA) M. tb H37Rv and low cytotoxicity. Compared to Delamanid, these new compounds (6, 7, 45) demonstrated dramatically improved physicochemical properties and are suitable for further in vitro and in vivo evaluation.