Search

Summary Background Market-applicable concurrent electrocardiogram (ECG) diagnosis for multiple heart abnormalities that covers a wide range of arrhythmias, with better-than-human accuracy, has not yet been developed. We therefore aimed to engineer a deep learning approach for the automated multilabel diagnosis of heart rhythm or conduction abnormalities by real-time ECG analysis. Methods We used a dataset of ECGs (standard 10 s, 12-channel format) from adult patients (aged ≥18 years), with 21 distinct rhythm classes, including most types of heart rhythm or conduction abnormalities, for the diagnosis of arrhythmias at multilabel level. The ECGs were collected from three campuses of Tongji Hospital (Huazhong University of Science and Technology, Wuhan, China) and annotated by cardiologists. We used these datasets to develop a convolutional neural network approach to generate diagnoses of arrythmias. We collected a test dataset of ECGs from a new group of patients not included in the training dataset. The test dataset was annotated by consensus of a committee of board-certified, actively practicing cardiologists. To evaluate the performance of the model we assessed the F1 score and the area under the curve (AUC) of the receiver operating characteristic (ROC) curve, as well as quantifying sensitivity and specificity. To validate our results, findings for the test dataset were compared with diagnoses made by 53 ECG physicians working in cardiology departments who had a wide range of experience in ECG interpretation (range 0 to >12 years). An external public validation dataset of 962 ECGs from other hospitals was used to study generalisability of the diagnostic model. Findings Our training and validation dataset comprised 180 112 ECGs from 70 692 patients, collected between Jan 1, 2012, and Apr 30, 2019. The test dataset comprised 828 ECGs corresponding to 828 new patients, recorded between Sept 11, 2012, and Aug 30, 2019. At the multilabel level, our deep learning approach to diagnosing heart abnormalities resulted in an exact match in 658 (80%) of 828 ECGs, exceeding the mean performance of physicians (552 [67%] for physicians with 0–6 years of experience; 571 [69%] for physicians with 7–12 years of experience; 621 [75%] for physicians with more than 12 years of experience). Our model had an overall mean F1 score of 0·887 compared with 0·789 for physicians with 0–6 years of experience, 0·815 for physicians with 7–12 years of experience, and 0·831 for physicians with more than 12 years of experience. The model had a mean AUC ROC score of 0·983 (95% CI 0·980–0·986), sensitivity of 0·867 (0·849–0·885) and specificity of 0·995 (0·994–0·996). Promising F1 scores were also obtained from the external public database using our proposed model without any model modifications (mean F1 scores of 0·845 in multilabel and 0·852 in single-label ECGs). Interpretation Our model is more accurate than physicians working in cardiology departments at distinguishing a range of distinct arrhythmias in single-label and multilabel ECGs, laying a promising foundation for computational decision-support systems in clinical applications. Funding National Natural Science Foundation of China and Hubei Science and Technology Project.

Cytogenetic abnormalities are powerful prognostic factors in multiple myeloma (MM) and are routinely analyzed by FISH on bone marrow (BM) plasma cells (PC). Although considered the gold standard, FISH experiments can be laborious and expensive. Therefore, array-CGH (aCGH) has been introduced as an alternative approach for detecting copy number aberrations (CNA), reducing the number of FISH experiments per case and yielding genome-wide information. Currently, next generation sequencing (NGS) technologies offer new perspectives for the diagnostic workup of malignant disorders. In this study, we examined ultra-low depth whole genome sequencing (LDS) as a valid alternative for aCGH for the detection of CNA in BM PC in MM. To this end, BM aspirates obtained in a diagnostic setting from 20 MM cases were analyzed. CD138+ cell-sorted samples were subjected to FISH analysis. DNA was extracted for subsequent aCGH and LDS analysis. CNA were detected by aCGH and LDS in all but one case. Importantly, all CNA identified by parallel first generation aCGH analysis were also detected by LDS, along with six additional CNA in five cases. One of these additional aberrations was in a region of prognostic importance in MM and was confirmed using FISH. However, risk stratification in these particular cases was unaffected. Thus, a perfectly concordant prognostication between array-CGH and LDS was observed. This validates LDS as a novel and cost-efficient tool for the detection of CNA in MM. ispartof: GENES CHROMOSOMES & CANCER vol:59 issue:8 pages:465-471 ispartof: location:United States status: published

Primary biliary cholangitis (PBC) is an autoimmune liver disease featured with bile duct injury, ductopenia and proliferation, periportal inflammation and fibrosis. Clinical manifestations of PBC vary from almost no symptoms to different degrees of pruritus plus symptoms of liver dysfunction. This review intends to update our understanding in the mechanisms of hepatic fibrogenesis under chronic biliary injury. Underlying mechanisms are proposed for a better understanding and more effective therapeutic targets. With genetic predisposition, lesions from bile ducts cause damage to biliary epithelial cells (BECs); and simultaneous autoimmunity reinforces a vicious cycle of BEC damage, inflammatory infiltration, BEC proliferation and fibrogenic responses. Therapeutic efficacy of immunosuppressive agents has been unsatisfactory. As a major variable in PBC progression, fibrosis has been paid a great deal of attention but has not been responsive to various treatments.

The neutrophil-to-lymphocyte ratio (NLR) has been proved to be a strong predictor of carotid atherosclerotic plaque, but the correlation between NLR and the stability of carotid plaque is not clear. Thus we conducted a study to evaluate the correlation between NLR and the stability of carotid atherosclerotic plaque, and to develop a new evaluation scale for rapid clinical evaluation of carotid plaque stability.We recruited 528 patients with acute anterior circulation ischemic stroke who were in accordance with extracranial and intracranial large artery atherosclerosis of Chinese ischemic stroke subtype. Blood routine examination and carotid ultrasound examination were performed on admission. According to the ultrasonic characteristics, the patients were divided into plaque stabilization group and plaque instability group.There was significant difference in NLR between plaque stability and instability groups (P 0.001). The risk of plaque instability increased with the increase of NLR (odds ratio (OR), 4.737; 95% confidence interval (CI), 3.404-6.592; P 0.001). Receiver operating characteristic (ROC) curve showed that the critical point of NLR is 2.55 and the area under the curve (AUC) was 0.782 (95%CI, 0.740-0.823; P 0.001). The best cut-off value of the evaluation scale was ≥ 4 points (sensitivity, 0.77; specificity, 0.75; accuracy, 0.76).There is a correlation between NLR and carotid plaque instability. NLR may be useful as a potential inflammation biomarker indicating the risk of unstable carotid plaques. The new scoring scale is a reliable index to predict the stability of carotid plaque.

STUDY QUESTION: Can genome-wide haplotyping increase success following preimplantation genetic testing for a monogenic disorder (PGT-M) by including zygotes with absence of pronuclei (0PN) or the presence of only one pronucleus (1PN)? SUMMARY ANSWER: Genome-wide haplotyping 0PNs and 1PNs increases the number of PGT-M cycles reaching embryo transfer (ET) by 81% and the live-birth rate by 75%. WHAT IS KNOWN ALREADY: Although a significant subset of 0PN and 1PN zygotes can develop into balanced, diploid and developmentally competent embryos, they are usually discarded because parental diploidy detection is not part of the routine work-up of PGT-M. STUDY DESIGN, SIZE, DURATION: This prospective cohort study evaluated the pronuclear number in 2229 zygotes from 2337 injected metaphase II (MII) oocytes in 268 cycles. PGT-M for 0PN and 1PN embryos developing into Day 5/6 blastocysts with adequate quality for vitrification was performed in 42 of the 268 cycles (15.7%). In these 42 cycles, we genome-wide haplotyped 216 good quality embryos corresponding to 49 0PNs, 15 1PNs and 152 2PNs. The reported outcomes include parental contribution to embryonic ploidy, embryonic aneuploidy, genetic diagnosis for the monogenic disorder, cycles reaching ETs, pregnancy and live birth rates (LBR) for unaffected offspring. PARTICIPANTS/MATERIALS, SETTING, METHODS: Blastomere DNA was whole-genome amplified and hybridized on the Illumina Human CytoSNP12V2.1.1 BeadChip arrays. Subsequently, genome-wide haplotyping and copy-number profiling was applied to investigate the embryonic genome architecture. Bi-parental, unaffected embryos were transferred regardless of their initial zygotic PN score. MAIN RESULTS AND THE ROLE OF CHANCE: A staggering 75.51% of 0PN and 42.86% of 1PN blastocysts are diploid bi-parental allowing accurate genetic diagnosis for the monogenic disorder. In total, 31% (13/42) of the PGT-M cycles reached ET or could repeat ET with an unaffected 0PN or 1PN embryo. The LBR per initiated cycle increased from 9.52 to 16.67%. LIMITATIONS, REASONS FOR CAUTION: The clinical efficacy of the routine inclusion of 0PN and 1PN zygotes in PGT-M cycles should be confirmed in larger cohorts from multicenter studies. WIDER IMPLICATIONS OF THE FINDINGS: Genome-wide haplotyping allows the inclusion of 0PN and 1PN embryos and subsequently increases the cycles reaching ET following PGT-M and potentially PGT for aneuploidy (PGT-A) and chromosomal structural rearrangements (PGT-SR). Establishing measures of clinical efficacy could lead to an update of the ESHRE guidelines which advise against the use of these zygotes. STUDY FUNDING/COMPETING INTEREST(S): SymBioSys (PFV/10/016 and C1/018 to J.R.V. and T.V.), the Horizon 2020 WIDENLIFE: 692065 to J.R.V., T.V., E.D., A.D. and M.Z.E. M.Z.E., T.V. and J.R.V. co-invented haplarithmisis ('Haplotyping and copy-number typing using polymorphic variant allelic frequencies'), which has been licensed to Agilent Technologies. H.M. is fully supported by the (FWO) (ZKD1543-ASP/16). The authors have no competing interests to declare. ispartof: HUMAN REPRODUCTION vol:33 issue:12 pages:2302-2311 ispartof: location:England status: published

Background: To assess the feasibility of radiomics based on precontrast MRI for the distinguish of s-HCC and pre-HCC.Method: We retrospectively analyzed 146 nodules from 78 patients, with pathological confirmed. Each nodule was segment on precontrast MRI sequence(TIWI and fat-suppression T2WI), retrospectively. 1223radiomics features were extracted and the optimal 10 features were selected by LASSO to establish the logistic regression radiomics model. Result: The AUC, sensitivity and specificity of the training group and test group were 0.757 (95% CI 0.638 -0.853), 83.02% , 66.67% and 0.789 (95% CI 0.643-0.895), 88.89% and 80.00%, respectively. The AUC, sensitivity and specificity of the training group and test group were 0.903 (95% CI 0.807-0.962), 86.79% , 86.67% and 0.778 (95% CI 0.632-0.887), 75.00%, 80.00%, respectively. Delong test has proved that, the diagnositic performances of radiomics model based on T2WI were higher than that of radiomics model based on T1WI (p = 0.0379).Conclusion: Radiomics model can classify s-HCC and pre-HCC based on precontrast MRI. And may serve as an adjunct tool for accurate diagnosis of s-HCC.

Chromosome instability is inherent to human IVF embryos, but the full spectrum and developmental fate of chromosome anomalies remain uncharacterized. Using haplotyping-based preimplantation genetic testing for monogenic diseases (PGT-M), we mapped the parental and mechanistic origin of common and rare genomic abnormalities in 2300 cleavage stage and 361 trophectoderm biopsies. We show that while single whole chromosome aneuploidy arises due to chromosome-specific meiotic errors in the oocyte, segmental imbalances predominantly affect paternal chromosomes, implicating sperm DNA damage in segmental aneuploidy formation. We also show that postzygotic aneuploidy affects multiple chromosomes across the genome and does not discriminate between parental homologs. In addition, 6% of cleavage stage embryos demonstrated signatures of tripolar cell division with excessive chromosome loss, however hypodiploid blastomeres can be excluded from further embryo development. This observation supports the selective-pressure hypothesis in embryos. Finally, considering that ploidy violations may constitute a significant proportion of non-viable embryos, using haplotyping-based approach to map these events might further improve IVF success rate. ispartof: NPJ GENOMIC MEDICINE vol:6 issue:1 ispartof: location:England status: published

Background: Most gastric cancer (GC) patients are diagnosed at an advanced stage, which indicates their poor prognosis. We aimed to investigate novel fecal microbial markers for clinical application in the screening and early diagnosis of gastric cancer (GC). Methods: This is an observational study from three cohorts. In discovery cohort, 16S rRNA gene analysis was performed in paired samples (gastric tissues, feces) from GC and CG patients to determine differential microbes. The relative abundances of the differential microbes were detected using qPCR in training cohort. The diagnostic efficacy of fecal marker Sa/Sc was validated in validation cohort. Findings: 575 gastric cancer (GC) patients, 468 chronic gastritis (CG) patients were enrolled from ten hospitals in China. Significant enrichments of Streptococcus anginosus (Sa) and Streptococcus constellatus (Sc) in both cancerous tissues (P < 0·05) and feces ( P < 0·0001) were observed in patients with intraepithelial neoplasia, early and advanced GC. Either the marker parallel test Sa ∪ Sc or single marker Sa/Sc demonstrated superior sensitivity (Sa: 75·6% vs. 72·1%, P

Hedgehog (Hh) signaling is a classic morphogen in controlling embryonic development and tissue repairing. Aberrant activation of Hh signaling has been well documented in liver cancer, including hepatoblastoma, hepatocellular carcinoma (HCC) and cholangiocarcinoma. The present review aims to update the current understanding on how abnormal Hh signaling molecules modulate initiation, progression, drug resistance and metastasis of HCC. The latest relevant literature was reviewed with our recent findings to provide an overview regarding the molecular interplay and clinical relevance of the Hh signaling in HCC management. Hh signaling molecules are involved in the transformation of pre-carcinogenic lesions to malignant features in chronic liver injury, such as nonalcoholic steatohepatitis. Activation of GLI target genes, such as ABCC1 and TAP1, is responsible for drug resistance in hepatoma cells, with a CD133−/EpCAM− surface molecular profile, and GLI1 and truncated GLI1 account for the metastatic feature of the hepatoma cells, with upregulation of matrix metalloproteinases. A novel bioassay for the Sonic Hh ligand in tissue specimens may assist HCC diagnosis with negative α-fetoprotein and predict early microvascular invasion. In-depth exploration of the Hh signaling deepens our understanding of its molecular modulation in HCC initiation, drug sensitivity and metastasis, and guides precise management of HCC on an individual basis.

Ultrasound segmentation models provide powerful tools for the diagnosis process of ultrasound examinations. However, developing such models for ultrasound videos requires densely annotated segmentation masks of all frames in a dataset, which is unpractical and unaffordable. Therefore, we propose a weakly-supervised learning (WSL) approach to accomplish the goal of video-based ultrasound segmentation. By only annotating the location of the start and end frames of the lesions, we obtain frame-level binary labels for WSL. We design Video Co-Attention Network to learn the correspondence between frames, where CAM and co-CAM will be obtained to perform lesion localization. Moreover, we find that the essential factor to the success of extracting video-level information is applying our proposed consistency regularization between CAM and co-CAM. Our method achieves an mIoU score of 45.43% in the breast ultrasound dataset, which significantly outperforms the baseline methods. The codes of our models will be released.

Proximal to distal Y stenting technique is a modified bifurcation technique based on the original Y stenting technique described over 20 years ago. We use a bench top model to illustrate the steps of the technique, which can provide both provisional and full coverage options, using radial artery access. This technique may be applied in clinical settings on a wide range of bifurcation anatomies with a number of unique advantages.

To evaluate the plasma levels and clearance of procalcitonin (PCTc) as prognostic biomarkers for patients with abdominal sepsis. A retrospective study of 103 patients with abdominal sepsis was conducted in our intensive care unit (ICU) from 2016 to 2018. Procalcitonin (PCT) plasma levels were measured at the time of diagnosis of abdominal sepsis and daily over the next 5 days. PCTc was calculated from day 3 to day 5. The prognostic accuracy of PCTc was expressed as the area under the receiver operating characteristic curve (AUROC). The Kaplan–Meier method was used to compare the survival curves by log-rank test. Logistic regression analysis was used to predict the 28-day mortality. The primary outcome was 28-day mortality since admission to ICU. Serum PCT levels from day 1 to day 5 did not differ significantly between survivors and non-survivors, while PCTc on day 5 was statistically significantly higher in survivors than that in non-survivors (86.4% vs 38.7%, P 70% compared with those with PCTc-day 5

A couple of molecular-targeting medications, such as Lenvatinib, are available for the treatment of hepatocellular carcinoma (HCC) in addition to Sorafenib in an advanced stage. Approval for the use of immune check-point inhibitors, such as Nivolumab and Pembrolizumab has shifted the paradigm of current HCC treatment, and the monotherapy or in combination with Lenvatinib or Sorafenib has significantly extended overall survival or progression-free survival in a large portion of patients. A combination of programmed cell death ligand-1 (PD-L1) inhibitor Atezolizumab with a vascular endothelial growth factor (VEGF) inhibitor, Bevacizumab, has recently achieved promising outcome in unresectable HCC patients. Other immunotherapy, such as chimeric antigen receptor T (CAR-T) cell therapy has achieved an evolutional success in hematologic malignancies, and has extended its use in deadly solid tumors, such as HCC. Although there exist various barriers, novel approaches are developed to move potential adoptive T cell therapy strategies, including cytokine-induced killer (CIK) cells, tumor-infiltrating lymphocytes (TIL), T cell receptor (TCR) T cells, CAR-T cells, to clinical application.

Background: To study the endoscopic trans-lateral molar (ETLM) approach to infratemporal fossa (ITF) lesions and analyze the advantages and disadvantages of this method. Methods: Four cases of ITF lesions were analyzed retrospectively. The clinical features, diagnosis and treatments, the operative process, and clinical applications of this surgical approach were discussed. Results: Postoperative pathologies were 2 pleomorphic adenomas, 1 schwannoma, and 1 inflammatory lesion. All patients had self-resolving cheek swelling and pharyngalgia in the short term, but 2 patients had numbness in the long term. There was no infection or bleeding in the postoperative period, and no difficulty in chewing after disease recovery. There was no tumor recurrence during the follow-up period. Conclusion: The ETLM approach is convenient, minimally invasive, and allows complete excision of benign ITF lesions, posterior to the lateral pterygoid muscle and mainly below the level of the hard palate. It is a simple and direct access to the ITF, but it is a narrow access because of the limitations of bones and soft tissues. Appropriate patient selection is mandatory for successful surgery.

Background: Pancreatic ductal adenocarcinoma is a common malignancy with high morbidity. MicroRNAs have been demonstrated to be critical posttranscriptional regulators in tumorigenesis. This study aimed to investigate the effect of microRNA-590 on the proliferation and apoptosis of pancreatic ductal adenocarcinoma. Material and Methods: The expression of microRNA-590 and high mobility group AT-hook 2 were examined in clinical pancreatic ductal adenocarcinoma tissues. Pancreatic ductal adenocarcinoma cell line Capan-2 was employed and transfected with microRNA-590 mimics or inhibitor. The correlation between microRNA-590 and high mobility group AT-hook 2 was verified by luciferase reporter assay. Cell viability and apoptosis were detected by MTT and flow cytometry assay. The protein level of high mobility group AT-hook 2, AKT, p-AKT, mTOR, and phosphorylated mTOR were analyzed by Western blotting. Results: MicroRNA-590 was found to be negatively correlated with the expression of high mobility group AT-hook 2 in pancreatic ductal adenocarcinoma tissues. Further studies identified high mobility group AT-hook 2 as a direct target of microRNA-590. Moreover, overexpression of microRNA-590 downregulated expression of high mobility group AT-hook 2, reduced cell viability, and promoted cell apoptosis, while knockdown of miR-590 led to an inverse result. MicroRNA-590 also suppressed the phosphorylation of AKT and mTOR without altering total AKT and mTOR levels. Conclusion: Our study indicated that microRNA-590 negatively regulates the expression of high mobility group AT-hook 2 in clinical specimens and in vitro. MicroRNA-590 can inhibit cell proliferation and induce cell apoptosis in pancreatic ductal adenocarcinoma cells. This regulatory effect of microRNA-590 may be associated with AKT signaling pathway. Therefore, microRNA-590 has the potential to be used as a biomarker for predicting the progression of pancreatic ductal adenocarcinoma.

Background: Electrocardiography (ECG) is a fundamental diagnostic tool, frequently used in clinical practice, for testing heart conditions. Recently, computer-aided ECG interpretation has become more widely investigated. However, researches on concurrent ECG diagnosis for multiple heart abnormalities are limited and therefore a market-applicable multi-label automatic diagnosis framework that covers a wide range of arrhythmias, with better-than-human accuracy in diagnosing ECGs with multi-labels, has not yet been developed. We therefore aimed to engineer a deep learning approach for the automated multi-label diagnosis of impulse or conduction abnormalities by realtime ECG analysis. Methods: We utilized a dataset with 70,692 patients, consisting of adults with age ≥18 years. 180112 ECGs (standard 10-second and 12-lead format) with 21 distinct rhythm classes, including various impulse or conduction abnormalities, were used for the diagnosis of arrhythmias in multi-label and single-label level. Base rhythm labels were assessed by trained personnel under the supervision of cardiologists. We ensured that each abnormality had a sample size of at least 4,000 ECGs, except idioventricular rhythm (1,780 ECGs due to the rarity of said disease), for compiling a balanced dataset. We allocated ECGs to a training or a validation dataset for the multi-label diagnostic model training, and evaluated model performance on a test dataset by calculating the area under the curve (AUC) of the receiver operating characteristic (ROC) curve, as well as quantifying accuracy, sensitivity, specificity, and the F1 score. The test dataset was annotated by the consensus of a committee of board-certified actively-practicing cardiologists. Validation of results for the test dataset were compared to diagnoses made by 53 ECG cardiologists who have a wide range of ECG interpretation experience (range=0-12+ years). Findings: We demonstrated that in multi-label level, our convolutional neural network (CNN) approach toward diagnosing heart abnormalities resulted in AUC scores above 0.913 for most of the diseases, with an average score of 0.985 (95% confidence interval (CI) 0.988-0.994). The model obtained an average F1 score of 0.890 (95% CI0.877- 0.903), sensitivity of 0.869 (95% CI 0.851-0.887), specificity of 0.995 (95% CI 0.994- 0.996). The average F1 score computed from our deep learning model exceeded the score from human cardiologists regardless of ECG interpretation experience (0 12 years: 0.829). The CNN outputs also showed identical or increased performance with respect to average sensitivity and accuracy as compared to the cardiologists. In single-label level, the CNN achieved an average F1 of 0.918, higher than the average level of the cardiologists 0.798. Interpretation: Our automatic ECG diagnosis system is better than human cardiologists to classify particular waveforms into clinically defined categories accurately, to distinguish a wide range of distinct arrhythmias, and to apply both single label and multiple label ECGs. Our study is therefore a promising foundation for the deployment of computational decision-support systems in practical clinical applications. Funding Statement: Project supported by the Major International (Regional) Joint Research Project, the Ministry of Management Science, the National Natural Science Foundation of China (Grant No. 71520107003). Declaration of Interests: Dr. Yuan reports grants from National Natural Science Foundation of China, during the conduct of the study; In addition, Dr. Yuan has a patent An automatic ECG classification method, system and equipment based on deep learning algorithm pending to Ye Yuan, Xiaoyun Yang, Hongling Zhu, Yiran Wang, Cheng Cheng, Xingyi Li, Hang Yin, Jingyi Wang. There are no other conflicts of interest. Ethics Approval Statement: To protect patients’ privacy, we used anonymized data as to prevent breaches. Accordingly, a written informed consent was not required for this study, since the ECG samples were appropriately anonymized and de-identified, as according to the Health Insurance Portability and Accountability Act Safe Harbor provision. The study design was evaluated and exempted from full review by the Huazhong University of Science and Technology Institutional Review Board.

BACKGROUND Congenital diaphragmatic hernia (CDH) is characterized by a defective closure of the diaphragm occurring as an isolated defect in 60% of cases. Lung size, liver herniation, and pulmonary circulation are major prognostic indices. Isolated CDH genetics is heterogeneous and poorly understood. Whether genetic lesions are also outcome determinants has never been explored. OBJECTIVES To identify isolated CDH genetic causes, to fine map the mutational burden, and to search for a correlation between the genotype and the disease severity and outcome. METHODS Targeted massively parallel sequencing of 143 human and mouse CDH causative and candidate genes in a cohort of 120 fetuses with isolated CDH and detailed outcome measures. RESULTS Pathogenic and likely pathogenic variants were identified in 10% of the cohort. These variants affect both known CDH causative genes, namely, ZFPM2, GATA4, and NR2F2, and new genes, namely, TBX1, TBX5, GATA5, and PBX1. In addition, mutation burden analysis identified LBR, CTBP2, NSD1, MMP14, MYOD1, and EYA1 as candidate genes with enrichment in rare but predicted deleterious variants. No obvious correlation between the genotype and the phenotype or short-term outcome has been found. CONCLUSION Targeted resequencing identifies a genetic cause in 10% of isolated CDH and identifies new candidate genes.

Surfactin, a lipopeptide produced by Bacillus species, has been used for the oral delivery of insulin. In this study, another lipopeptide of iturin was tested for its ability to orally delivery insulin alone or plus surfactin. Iturin could form co-precipitate with insulin at acidic pH values. After treatment by ultrasonification, the structure of coprecipitate was destroyed that led to a significant decrease in hypoglycemic effect after oral administration. Iturin weakly binds to (Kd = 257 μM) and induce insulin structure more compact that is favorable for insulin uptake by the intestine. After being coated with Acryl-Eze by lyophilization, the coprecipitate formed the spherical enteric-coated insulin microparticles delivered by iturin with a relative oral bioavailability of 6.84% in diabetic mice. For further improving oral hypoglycemic effect, surfactin was added to form the spherical enteric-coated insulin microparticles in a formulation containing insulin, Acryl-Eze, iturin and surfactin at a ratio of 1:1:0.5: 0.5 (w/w), with an insulin encapsulation efficiency of 66.22%. The enteric-coated insulin microparticles delivered by iturin plus surfactin showed a classical profile for controlled release in the intestine with a relative bioavailability of 7.67% after oral administration, which could effectively control the postprandial blood glucose at a level about 50% of the initial one just like the subcutaneous injection. Collectively, iturin plus surfactin is more efficient for oral delivering insulin than the sole one, and the resultant enteric-coated insulin microparticles are potential for the development of oral insulin to control postprandial blood glucose in diabetic patients.

Surfactin is a Bacillus-produced natural lipopeptide, which can overcome the epithelial cell barriers for orally delivering insulin, but its ability to promote uptake of insulin by the intestine need to be further improved for a higher oral bioavailability. Here, we designed and synthesized several surfactin variants to improve its ability for oral delivery of insulin. Firstly, we replaced Glu with Gln in surfactin for decreasing its negative charges, but found this replacement weakened its ability to orally delivery insulin. We further chemically synthesized surfactin variant by replacing its fatty acid chain (C15) with a shortened one (C14), and found this replacement did not influence its ability to orally deliver insulin. Lastly, we replaced its amino acids (Leu) with more hydrophobic ones (Ile), and found the replacement could significantly improve its ability to deliver insulin, with a maximal blood glucose decrease to 27.33% of the initial level and an insulin bioavailability of 18.25%. We also replaced its amino acids of Leu with Val, and Val with Ile, and found this replacement could also significantly improve its ability to deliver insulin with a maximal blood glucose decrease to 18.36% of the initial level and a high insulin bioavailability of 26.32% in diabetic mice. Further analysis by CD, we found the surfactin variants with more hydrophobic amino acid residuals significantly induced insulin from rigid (α-helix) to flexible structure (β-sheet and random coil), that is favorable for insulin to permeate across the intestine epithelial membrane. Collectively, surfactin variants with more hydrophobicity are very potential for delivery of insulin in the everyday control of blood glucose.

We created Immense Surface Area Carbons (ISACs) by a novel heat treatment that stabilized the micelle structure in a biological based precursor prior to high temperature combined activation – pyrolysis. While displaying a morphology akin to that of commercial activated carbon, ISACs contain an unparalleled combination of electrochemically active surface area and pore volume (up to 4051 m2 g−1, total pore volume 2.60 cm3 g−1, 76% small mesopores). The carbons also possess the benefit of being quite pure (combined O and N: 2.6–4.1 at%), thus allowing for a capacitive response that is primarily EDLC. Tested at commercial mass loadings (∼10 mg cm−2) ISACs demonstrate exceptional specific capacitance values throughout the entire relevant current density regime, with superior rate capability primarily due to the large fraction of mesopores. In the optimized ISAC, the specific capacitance (Cg) is 540 F g−1 at 0.2 A g−1, 409 F g−1 at 1 A g−1 and 226 F g−1 at a very high current density of 300 A g−1 (∼0.15 second charge time). At intermediate and high currents, such capacitance values have not been previously reported for any carbon. Tested with a stable 1.8 V window in a 1 M Li2SO4 electrolyte, a symmetric supercapacitor cell yields a flat energy–power profile that is fully competitive with those of organic electrolyte systems: 29 W h kg−1 at 442 W kg−1 and 17 W h kg−1 at 3940 W kg−1. The cyclability of symmetric ISAC cells is also exceptional due to the minimization of faradaic reactions on the carbon surface, with 80% capacitance retention over 100 000 cycles in 1 M Li2SO4 and 75 000 cycles in 6 M KOH.

Heterostructured ZnS/InP nanowires, composed of single-crystalline ZnS nanowires coated with a layer of InP shell, were synthesized via a one-step chemical vapor deposition process. As-grown heterostructured ZnS/InP nanowires exhibited an ultrahigh Ion/Ioff ratio of 4.91 × 103, a high photoconductive gain of 1.10 × 103, a high detectivity of 1.65 × 1013 Jones and high response speed even in the case of very weak ultraviolet light illumination (1.87 μW cm-2). The values are much higher than those of previously reported bare ZnS nanowires owing to the formation of core/shell heterostructures. Flexible ultraviolet photodetectors were also fabricated with the heterostructured ZnS/InP nanowires, which showed excellent mechanical flexibility, electrical stability and folding endurance besides excellent photoresponse properties. The results elucidated that the heterostructured ZnS/InP nanowires could find good applications in next generation flexible optoelectronic devices.

Effects of propofol, midazolam and dexmedetomidine on patients with sepsis in intensive care unit (ICU) and on arterial blood gas (ABG) were studied. In total 429 ICU patients with sepsis, admitted to Renji Hospital, School of Medicine, Shanghai Jiaotong University from May 2015 to January 2019, were selected as research subjects for a prospective analysis. All patients received basic treatment, such as anti-infection treatment, correction of shock and improvement of microcirculation. One hundred and fifty-two patients who were treated with propofol for sedation served as group A, 146 patients who were treated with midazolam for sedation served as group B, and 131 patients who were treated with dexmedetomidine for sedation served as group C. The three groups of patients were compared in terms of diastolic blood pressure (DBP), systolic blood pressure (SBP), heart rate (HR), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), cardiac troponin T (cTnT) and creatine kinase-MB (CK-MB) before and after treatment. APACHE II score was used to evaluate the sedative effects. The wake-up time of the patients, the length of ICU stay and the adverse reactions were recorded. There was no significant difference among groups A, B and C in terms of HR, SBP, DBP, PaO2, PaCO2, cTnT, CK-MB and APACHE II score before treatment, and SBP, DBP, cTnT and HR after treatment (P>0.050). After treatment, there was no significant difference between groups A and B with respect to CK-MB and APACHE II score (P>0.050). The wake-up time in group A was significantly longer than that in groups B and C (P

Multidrug resistance is one of the reasons for low survival of advanced hepatocellular carcinoma (HCC). Our previous studies indicate that the hedgehog signalling is involved in hepatic carcinogenesis, metastasis and chemo‐resistance. The present study aims to uncover molecular mechanisms underlying hepatoma chemo‐resistance. TAP1 and GLI1/2 gene expression was assessed in both poorly differentiated hepatoma cells and HCC specimens. Potential GLI‐binding site in the TAP1 promoter sequence was validated by molecular assays. Approximately 75% HCC specimens exhibited an elevated expression of hedgehog GLI1 transcription factor compared with adjacent liver tissue. Both GLI1/2 and TAP1 protein levels were significantly elevated in poorly differentiated hepatoma cells. Both Huh‐7‐trans and Huh‐7‐DN displayed more karyotypic abnormalities and differential gene expression profiles than their native Huh‐7 cells. Sensitivity to Sorafenib, doxorubicin and cisplatin was remarkably improved after either GLI1 or TAP1 gene was inhibited by an RNAi approach or by a specific GLI1/2 inhibitor, GANT61. Further experiments confirmed that hedgehog transcription factor GLI1/2 binds to the TAP1 promoter, indicating that TAP1 is one of GLI1/2 target genes. In conclusion, TAP1 is under direct transcriptional control of the hedgehog signalling. Targeting hedgehog signalling confers a novel insight into alleviating drug resistance in the treatment of refractory HCC.

STUDY QUESTION Is it possible to haplotype parents using parental siblings to leverage preimplantation genetic testing (PGT) for monogenic diseases and aneuploidy (comprehensive PGT) by genome-wide haplotyping? SUMMARY ANSWER We imputed identity-by-state (IBS) sharing of parental siblings to phase parental genotypes. WHAT IS KNOWN ALREADY Genome-wide haplotyping of preimplantation embryos is being implemented as a generic approach for genetic diagnosis of inherited single-gene disorders. To enable the phasing of genotypes into haplotypes, genotyping the direct family members of the prospective parent carrying the mutation is required. Current approaches require genotypes of either (i) both or one of the parents of the affected prospective parent or (ii) an affected or an unaffected child of the couple. However, this approach cannot be used when parents or children are not attainable, prompting an investigation into alternative phasing options. STUDY DESIGN, SIZE, DURATION This is a retrospective validation study, which applied IBS-based phasing of parental haplotypes in 56 embryos derived from 12 PGT families. Genome-wide haplotypes and copy number profiles generated for each embryo using the new phasing approach were compared with the reference PGT method to evaluate the diagnostic concordance. PARTICIPANTS/MATERIALS, SETTING, METHODS This study included 12 couples with a known hereditary genetic disorder, participating in the comprehensive PGT program and with at least one parental sibling available (e.g. brother and/or sister). Genotyping data from both prospective parents and the parental sibling(s) were used to perform IBS-based phasing and to trace the disease-associated alleles. The outcome of the IBS-based PGT was compared with the results of the clinically implemented reference haplotyping-based PGT method. MAIN RESULTS AND THE ROLE OF CHANCE IBS-based haplotyping was performed for 12 PGT families. In accordance with the theoretical prediction of allele sharing between sibling pairs, 6 out of 12 (50%) couples or 23 out of 56 embryos could be phased using parental siblings. In families where phasing was possible, haplotype calling in the locus of interest was 100% concordant between the reference PGT method and IBS-based approach using parental siblings. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION Phasing of parental haplotypes will only be possible when the disease locus lies in an informative region (categorized as IBS1). Phasing prospective parents using relatives with reduced genetic relatedness as a reference (e.g. siblings) decreases the size and the occurrence of informative IBS1 regions, necessary for haplotype calling. By including more than one extended family member, the chance of obtaining IBS1 coverage in the interrogated locus can be increased. A pre-PGT work-up can define whether the carrier couple could benefit from this approach. WIDER IMPLICATIONS OF THE FINDINGS Phasing by relatives extends the potential of comprehensive PGT, since it allows the inclusion of couples who do not have access to the standard phasing references, such as parents or offspring. STUDY FUNDING/COMPETING INTEREST(S) The study was funded by the KU Leuven grant (C14/18/092), Research Foundation Flanders (FWO; GA09311N), Horizon 2020 innovation programme (WIDENLIFE, 692065) and Agilent Technologies. J.R.V., T.V. and M.Z.E. are co-inventors of a patent ZL910050-PCT/EP2011/060211-WO/2011/157846 ‘Methods for haplotyping single-cells’ and ZL913096-PCT/EP2014/068315-WO/2015/028576 ‘Haplotyping and copy number typing using polymorphic variant allelic frequencies’ licensed to Agilent Technologies. The other authors have no conflict of interest to declare.

The purpose of this study was to focus on the underlying relationship between the hyperactivity for the peripheral monocytes and heat stroke by investigating the inflammatory oxidative activity of and the expression of superficial molecules. Peripheral blood samples were collected from 10 healthy adult volunteers. Human blood monocytes were isolated by density gradient centrifugation and sequent adherent culture. The objectives were divided into four groups: 43°C heat stress combined with lipopolysaccharide (LPS) group, 43°C heat stress group, LPS group, and control group. There were 10 cases in each group. An enzyme-linked immunosorbent assay (ELISA) test was used to measure the concentrations of supernatant inflammatory mediators (tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β) and interleukin-10 (IL-10)). After loaded by 2,7-Dichlorodi-hydrofluorescein-diacetate (DCFHDA) fluorescent probe, intracellular reactive oxygen species (ROS) levels were determined by a flow cytometry. After fluorescent microspheres incubation, the phagocytosis of monocytes was observed under a fluorescent microscope. Respectively, the flow cytometry and Western blot were used to evaluate the level of triggering receptor expressed on myeloid cells-1 (TREM-1) and Toll-like receptor-4 (TLR-4) on the monocytes. Furthermore, the mRNA expression of TREM-1 and TLR-4 was detected by real-time polymerase chain reaction (RT-PCR). The heat stress combined with LPS stimulation promoted the peripheral monocytes to produce inflammatory mediators (TNF-α, IL-1β, and IL-10) and release ROS. Otherwise, such complex strike significantly suppressed the phagocytic activity of monocytes in peripheral blood. Moreover, the expression of TREM-1, TLR-4 and CD86 was measured by the flow cytometry on peripheral monocytes which were respectively promoted by the union of heat stress and LPS. The results of Western blot and RT-PCR demonstrated the similar kinetics on these superficial molecules (TREM-1, TLR-4, and CD86) stimulated by the combination of heat stress and LPS. The underlying mechanism of the dysfunction for the peripheral monocytes may be related to the abnormal expression of superficial molecules TREM-1, TLR-4, and CD86 on the monocytes induced by heat stress and LPS.

The high expression of sonic hedgehog ligand (SHh) is closely correlated to the metastasis, drug resistance and poor prognosis of hepatocellular carcinoma (HCC). Therefore, sensitive, specific and efficient detection methods for SHh are needed for the early diagnosis and assessment of prognosis. Herein, an aptamer, AP32 that specifically binds to SHh (KD = 25.7 ± 4.1 nM) was obtained by SELEX technology with further optimization. In vivo experiments confirmed that AP32 has the potential to be an imaging probe for Huh-7 cell-derived xenograft. The interaction mode in 3-dimensional configuration between the aptamer and SHh was established by molecular simulation and confirmed by mutations at key sites of the aptamer. An aptasensor-based assay was successfully developed by conjugating Texas-Red-labeled AP32 to microbeads, and was used to analyze SHh content in hepatoma cell lysates, serum and HCC specimens. The method exhibited a broad detection range from 0.07 to 62.5 nM with a low detection limit of 69 pM, and a recovery rate of 104.6 ± 3.9% in serum. When the assay was used to measure SHh content in tissue lysates, the results demonstrated that it possessed 57.1% positivity, 100% specificity in distinguishing 28 HCC specimens from normal tissues, and was compensatory for detection of HCC in AFP-negative cases. Moreover, elevated SHh levels are indicative of portal vein invasion at 77.8% positive rate. This novel aptasensor-based SHh assay may offer a reliable means in predicting early metastasis and poor prognosis in hepatocellular carcinoma.

Bovine mastitis is an economical important microbial disease in dairy industry. Some recent human clinical trials have shown that oral probiotics supplementation could effectively control clinical mastitis, suggesting that the mechanism of mastitis protection might be achieved via the host gut microbiota. We aimed to test our hypothesis that bovine mastitis was related to changes in both the mammary and gut microbial profiles. By quantitative PCR, the milk and faecal microbial profiles of cows with low (5 cells/ml) and high (>1×106 cells/ml) somatic cell count (SCC) were compared. Firstly, we observed drastic differences in both the milk and faecal microbial compositions at genus and Lactobacillus-species levels between the two groups. Secondly, the pattern of faecal microbial community changes of mastitis cows was similar to that of the milk, characterised by a general increase in the mastitis pathogens (Enterococcus, Streptococcus and Staphylococcus) and deprivation of Lactobacillus and its members (L. salivarius, L. sakei, L. ruminis, L. delbrueckii, L. buchneri, and L. acidophilus). Thirdly, only the faecal lactobacilli, but not bifidobacteria correlated with the milk microbial communities and SCC. Our data together hint to a close association between bovine mastitis, the host gut and milk microbiota.

Previous studies demonstrated that tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) caused adverse effects on thyroid hormone (TH) imbalance in aquatic and avian organisms. This study focused on the effects of TDCIPP on thyroid function and hormone homeostasis in mammals. Pubertal female Sprague-Dawley rats were orally administered 50, 100, or 250 mg per kg per d of TDCIPP from postnatal day (PND) 22 to PND42 for 21 days. The serum triiodothyronine (T3) levels increased significantly at 250 mg per kg per d of TDCIPP. There were no significant differences in the body weight, serum thyroxine (T4) and free thyroxine (FT4) levels between the control and TDCIPP treated groups. There were significant dose-dependent increases in the mRNA and protein expression levels of genes related to drug metabolism (cytochrome-p450-3A1, CYP3A1) and TH clearance (udp-glucuronosyltransferase-1A6, UGT1A6) in the liver. Treatment with TDCIPP increased hepatic type 1 deiodinase (DIO1) mRNA at 250 mg per kg per d but down-regulated hepatic TH receptor beta (TRβ) mRNA expression. In addition, TDCIPP exposure induced slight thyroid follicular hyperplasia, and several genes involved in TH biosynthesis (NIS, TPO, Tg) were altered at 100 and 250 mg per kg per d of TDCIPP. Nevertheless, serum thyroid stimulating hormone (TSH) levels and the receptor (TSHr) mRNA significantly decreased at only the low dose group. Based on these results, we certified that TDCIPP disturbs the normal bioprocess on TH synthesis, biotransformation or clearance, and hepatic detoxification of pubertal female SD rats, causing thyroid function disorder.

Objective: To investigate the effects and mechanisms of p38 signaling pathway in pentagastrin-regulated cell proliferation of colorectal carcinoma cell line HT-29. Methods: HT-29 cell line of colorectal carcinoma was in vitro incubated and divided into the control group, pentagastrin group, proglumide group, and pentagastrin + proglumide group. MTT reduction assay was performed to detect the proliferation status of HT-29 cell line and determine the optimal dosage of pentagastrin and proglumide. Annexin V-fluorescein isothiocyanate flow cytometry was used to detect the proliferation index (PI) and apoptosis rate (AR) of HT-29 cells. Reverse transcriptase polymerase chain reaction was performed to detect the mRNA expression of the pentagastrin receptor/cholecystokinin-B receptor (CCK-BR) and p38. The protein and phosphorylation levels of p38 were estimated by western blotting. Results: RT-PCR detection showed that CCK-BR mRNA was expressed in the HT-29 cell line. Pentagatrin improved HT-29 cell proliferation in dosage of 6.25 - 100 mg/L, and the optimal dosage of pentagastrin was 25.0 mg/L. Proglumide had no significant effect on the proliferation of HT-29 cells, but significantly inhibited the proliferation of HT-29 cells stimulated by pentagastrin when the dosage of proglumide was 8.0 - 128.0 mg/L, and the optimal dosage was 32.0 mg/L. The AR in the pentagastrin group was significantly lower than that in the control group and in the pentagastrin + proglumide group. The PI in the pentagastrin group was significantly higher than that in the control group and in the pentagastrin + proglumide group. P38 phosphorylation level in the pentagastrin group was significantly lower than that in the control group, and in the pentagastrin + proglumide group. There were no significant differences in the mRNA and protein expression of p38 in the control, pentagastrin, proglumide and pentagastrin + proglumide groups. Conclusion: Pentagastrin can improve proliferation of the CRC cell line HT-29 and inhibit apoptosis via the p38 signal transduction pathway. This mechanism may be associated with suppressed p38 protein phosphorylation level due to inhibition of proglumide, a gastrin receptor antagonist.

BACKGROUND: It has been demonstrated in our previous study that poorly-differentiated hepatoma cells with low α-feto-protein (AFP) expression and a CD133-/EpCAM- surface profile exhibited aberrant hedgehog signaling (Hh) activity, epithelial-mesenchymal transition and chemoresistance. However, expression of canonical Hh molecules (Sonic ligand and SMO) was not increased in poorly-differentiated hepatoma cells in contrast to well-differentiated hepatoma cells, indicating that Hh transcription factors Gli-1/2 may be activated through a non-classic pathway. The present study aims to investigate whether the TGF-β/Smad3 pathway is responsible for Gli-2 activation through a non-canonical pathway. METHODS: Expression of Sonic Hh (Shh) and Smad3 was assessed in both hepatoma cells and HCC specimens. Gli-2 and Smad3 expression was suppressed by lentiviral vector-mediated RNA interference in poorly-differentiated hepatoma Huh-7-DN and HLE cells. Chemosensitivity was evaluated in hepatoma cells with a Gli-2 or Smad3 inhibitor. RESULTS: Significantly increased Gli-1 and Gli-2 protein levels were observed in 78% (11 out of 14) and 50% (7 out of 14) HCC specimens. In 7 HCC patients with a normal serum AFP value ( CONCLUSION: TGF-β/Smad3 activated Gli-2 and its direct transcriptional target ABCC1 in poorly-differentiated and chemoresistant hepatoma cells. Suppressing TGF-β/Smad3/Gli-2 signaling activity confers a potential molecular target in partially alleviating Sorafenib resistance for the treatment of refractory hepatoma. Citation Format: Jia Ding, Hui-yan Li, Li Zhang, Xiao-tian Zhou, Jian Wu. Non-canonical activation of Gli-2 by TGF-β/Smad3 signaling pathway in modulation of chemoresistance of hepatocellular carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 6336.

Antiangiogenic therapy can enhance the efficacy of chemotherapy against solid tumors. This study was to determine whether TT46, a potential VEGF vaccine from DNA shuffling with a helper T-cell epitope of tetanus toxin B subunit, could enhance the efficacy of doxorubicin to combat hepatocellular carcinoma (HCC). Compared with monotherapy, the combination with TT46 vaccination and doxorubicin could significantly reduce microvessel counts and inhibit tumor angiogenesis. Enhanced immunization with TT46 for total six doses could induce long-term response maintenance with high anti-VEGF antibody titers in body. As a result, the combination with enhanced TT46 vaccination and doxorubicin significantly inhibited the HCC growth, and improved the survival rate of HCC-bearing mice in both of subcutaneous tumor model and lung metastasis model. In conclusion, the combined therapy with TT46 vaccination and doxorubicin has antitumor effects in both a prophylactic and therapeutic setting in a mouse model of H22 hepatocellular carcinoma.

Non-invasive prenatal diagnosis for the common trisomies 21, 18 and 13 has become standard of care in many countries world-wide. We developed methods that enable not only the detection of those common trisomies but leverage the identification of all fetal and maternal genomic and chromosomal disorders. As of July 2017 NIPT is reimbursed for all pregnant women in Belgium, so the outcome of this screening program will be presented, as well as our innovative NIPT pipeline. Non-invasive prenatal diagnosis (NIPD) for monogenic diseases has only been tested on a small number of genes using a locus-specific approach and has not become widely implemented. Here, we present a new method that enables the deduction of fetal haplotype to detect inherited monogenic diseases and aneuploidy. Using targeted sequencing in a genome-wide level of resolution, parental haplotype inheritance in the fetus was determined non-invasively with high accuracy based on cfDNA haplotyping analysis. In all clinical cases, the haplotypes blocks linked with the wild-type allele were correctly identified, and a maternally inherited trisomy case was properly deduced. The method is generic and can deduce the presence or absence of any inherited disease allele. Moreover, it leverage the genome wide fetal haplotype at relatively low cost. Since genome wide haplotyping of embryos (karyomapping or OnePGT) enables the reconstruction of genome-wide haplotypes, copy-number profiles and are able to determine the segregational origin of haplotypes, those methodologies are being implemented as a generic method for preimplantation genetic testing (PGT) in many IVF laboratories. This non-invasive prenatal haplotyping approach can also confirm that the fetus is derived from the embryo transferred and will replace invasive prenatal confirmation testing.

Spiral ganglion neuron (SGN) damage and apoptosis can lead to noise-induced hearing loss, age-associated hearing loss and, in certain cases, auditory neuropathy. The apoptosis-inducing factor (AIF)-associated pathway may be important in this process. The present study aimed to investigate the expression levels of AIF and calpain in damaged SGNs. Glutamate (Glu) perfusion and cell culture in different concentrations of Glu were performed to damage the SGNs of Sprague-Dawley (SD) rats, with saline water used as a control Different concentrations (5, 10, 20 and 40 mM) of Glu were injected into the cochlear tympanic canal of 18 SD rats, and 10, 20 and 40 mM Glu were added to SGN cultures. Auditory brainstem responses (ABR) were measured prior to and 2 days following the injection of Glu. Immunofluorescent staining was used to detect the SGN damage and the expression levels of AIF and calpain in vivo and in in vitro. Transmission electron microscopy (TEM) was used to measure cell apoptosis and reverse transcription-quantitative polymerase chain reaction was used to analyse the gene expression levels of AIF and calpain in the damaged SGNs. The TEM identified mitochondrial vacuolisation, swelling of the SGN and hetero-chromatin formation. Injection of Glu reduced the number of SGNs and induced apoptosis. AIF was observed to translocate into the nuclei of the SGNs in the 20 and 40 mM Glu groups, and the expression levels of AIF and calpain were markedly upregulated in the modiolus of the Glu-damaged SGNs. The upregulation of AIF and calpain may be important in the process of SGN damage and apoptosis.

Hepatocellular carcinoma (HCC) is the third most lethal cancer and resistant to common chemotherapy. Tumor-initiating cells (T-ICs) that are thought to be responsible for tumorigenesis share surface markers and signaling pathways similar to normal tissue stem cells. Identification of T-ICs and elucidation of aberrant epigenetic modulation and self-renewal pathways may provide new insights into hepatic carcinogenesis, metastasis and chemotherapeutic resistance. Histone modification, DNA methylation and microenvironmental changes are considered as key elements to promote the derivation and function of T-ICs. In this review, we intend to compare the similarity and difference between normal liver stem cells and T-ICs, and to define the intrinsic and micro-environmental factors that lead to the transformation from normal liver stem cells to hepatic T-ICs. We believe that etiology, microenvironmental alteration, epigenetic modification and epithelial-mesenchymal transition play a fundamental role in initiating the transformation. Strategies targeting signaling molecules critical in modulating these processes may offer a personalized therapy for HCC in the future.

Cochlear progenitor cells are considered as one of the best candidates for hair cell regeneration, thus, the regulation of cochlear progenitor cell proliferation has become a focus in this field. Several genes expressed in the inner ear during postnatal development have been demonstrated to be involved in maintaining the proliferative potential of progenitor cells, but the mechanism for regulating the proliferation and differentiation of cochlear progenitor cells remains poorly understood. Telomerase reverse transcriptase (TERT) has rate limiting telomerase activity and the overexpression of TERT has been shown to promote cell proliferation in series of cell lines. The aim of the present study was to evaluate the expression of TERT in the postnatal development of the cochlea and progenitor cells. The results demonstrated that TERT was expressed in the basilar membranes during the first postnatal week. In vitro, TERT expression in progenitor cells reached a maximum at day 4 after culture and decreased as the culture time prolonged or the cell passage number increased. These results led us to hypothesize that TERT may be involved in the development of the cochlea and in maintaining the proliferation ability of progenitor cells.

Acrolein is a ubiquitous dietary and environmental pollutant, which can also be generated endogenously during cellular stress. However, the molecular mechanisms underlying acrolein-induced neurotoxicity, especially in ototoxicity conditions, have not been fully determined. In this study, we investigated the mechanisms on acrolein-induced toxicity in primary cultured cochlear nucleus neurons with focus on Sirt3, a mitochondrial deacetylase. We found that acrolein treatment induced neuronal injury and programmed cell death (PCD) in a dose dependent manner in cochlear nucleus neurons, which was accompanied by increased intracellular reactive oxygen species (ROS) generation and lipid peroxidation. Acrolein exposure also significantly reduced the mitochondrial membrane potential (MMP) levels, promoted cytochrome c release and decreased mitochondrial ATP production. In addition, increased ER tracker fluorescence and activation of ER stress factors were observed after acrolein treatment, and the ER stress inhibitors were shown to attenuate acrolein-induced toxicity in cochlear nucleus neurons. The results of western blot and RT-PCR showed that acrolein markedly decreased the expression of Sirt3 at both mRNA and protein levels, and reduced the activity of downstream mitochondrial enzymes. Furthermore, overexpression of Sirt3 by lentivirus transfection partially prevented acrolein-induced neuronal injury in cochlear nucleus neurons. These results demonstrated that acrolein induces mitochondrial dysfunction and ER stress in cochlear nucleus neurons, and Sirt3 acts as an endogenous protective factor in acrolein-induced ototoxicity.

Aim: Development of EGF and VEGF vaccines with high antigenicity for combined immunotherapy of EGF–EGFR signaling-dependent epithelial tumors such as breast cancer. Method: EGF genes from mouse, human and chicken were randomly assembled to chimeric genes by DNA shuffling, then a chimeric EGF was selected out by PCR, SDS-PAGE and immunization for combined immunotherapy of breast cancer with a previously constructed chimeric VEGF vaccine from shuffling. Results: Combined vaccination with chimeric EGF and VEGF from shuffling could induce high titer of antibodies against EGF and VEGF to inhibit tumor growth and angiogenesis, and improve the survival rate of mice with breast cancer. Conclusion: Combined vaccination with EGF and VEGF from shuffling showed better immunotherapy on EGF–EGFR signaling-dependent epithelial tumors such as breast cancer than the single-agent EGF vaccination.

Early detection of pulmonary cancer is the most promising way to enhance a patient’s chance for survival. Accurate pulmonary nodule detection in computed tomography (CT) images is a crucial step in diagnosing pulmonary cancer. In this paper, inspired by the successful use of deep convolutional neural networks (DCNNs) in natural image recognition, we propose a novel pulmonary nodule detection approach based on DCNNs. We first introduce a deconvolutional structure to Faster Region-based Convolutional Neural Network (Faster R-CNN) for candidate detection on axial slices. Then, a three-dimensional DCNN is presented for the subsequent false positive reduction. Experimental results of the LUng Nodule Analysis 2016 (LUNA16) Challenge demonstrate the superior detection performance of the proposed approach on nodule detection (average FROC-score of 0.893, ranking the 1st place over all submitted results), which outperforms the best result on the leaderboard of the LUNA16 Challenge (average FROC-score of 0.864).

Spiral ganglion neuron (SGN) injury is a generally accepted precursor of auditory neuropathy. Receptor-interacting protein 3 (RIP3) has been reported as an important necroptosis pathway mediator that can be blocked by necrostatin-1 (Nec-1). In our study, we sought to identify whether necroptosis participated in SGN injury. Ouabain was applied to establish an SGN injury model. We measured the auditory brain-stem response (ABR) threshold shift as an indicator of the auditory conditions. Positive β3-tubulin immunofluorescence staining indicated the surviving SGNs. RIP3 expression was evaluated using immunofluorescence, quantitative real-time polymerase chain reaction and western blot. SGN injury promoted an increase in RIP3 expression that could be suppressed by application of the necroptosis inhibitor Nec-1. A decreased ABR threshold shift and increased SGN density were observed when Nec-1 was administered with apoptosis inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD). These results demonstrated that necroptosis is an indispensable pathway separately from apoptosis leading to SGN death pathway, in which RIP3 plays an important role.

A novel and sensitive method has been successfully developed for quantitative detection of invA gene of Salmonella. The surface of sensor chip was modified and then target sequence and probe-2 were induced, which formed a sandwich structure. Under optimal conditions, the SPR-based DNA sensor displayed a linear range of 200 pM - 10 nM and a low detection limit of 200 pM for synthetic target sequence. The excellent sensitivity and high stability of the designed biosensor, make this strategy a promising tool for Salmonella screening.

Introduction Genome-wide haplotyping strategies, such as karyomapping, haplarithmisis and OnePGT, have paved the way for comprehensive PGT, leveraging PGT-M, PGT-A and PGT-SR in a single workflow. Nevertheless, family-specific challenges can complicate the implementation of current methods. First, to enable haplotype reconstruction, genotyping the prospective parents and direct family members, i.e. offspring of the couple or their parent(s), is required. However, if these references are not available or suitable for phasing, such approaches cannot be used, prompting investigation towards alternative phasing modules. Additionally, the advancement of current technologies leads to the identification of an increasing number of variants of unknown significance. Development of novel analysis approaches can assist further evaluation and interpretation of such variants, based on the familial segregation of the variant, in order to assess if PGT is an appropriate clinical option. Material & methods Twelve couples i) burdened with a known hereditary genetic disorder participating in the genome-wide haplotyping-based PGT-M program and ii) with at least one parental sibling from the parental side carrying the genetic disorder available, were recruited for the development of the new method. Genotyping data from both prospective parents and parental sibling(s) were used to trace the shared and/or different allele and impute the disease-carrying allele based on identity-by-state (IBS) information. Concurrent haplotyping and copy number typing of embryo biopsies was then performed using an adapted version of siCHILD/haplarithmisis. Results Six out of 12 (50%) PGT-M couples could be phased using parental sibling(s). This is in line with the theoretical expectations of allele-sharing between sibling-pairs. Genome-wide haplotypes and copy-number profiles generated for each embryo using the new phasing approach were consequently compared to the clinical results, showing 100% concordancy. Clinical implementation of the method has resulted in the analysis of 7 families so far. Additionally, IBS analysis has enabled the inclusion of two special cases in the PGT program. In the first, PGT-M was offered for an X-linked disorder. The female partner had karyotype 47,XXX with presence of both maternal X chromosomes, so that no distinction could be made between the affected and unaffected allele. In the second, IBS analysis revealed the de novo and consequently pathogenic nature of an autosomal dominant variant in the maternal grandmother, allowing PGT. Conclusions IBS analysis in the context of PGT can be proven to be advantageous on several levels. First, it allows offering genome-wide linkage analysis-based comprehensive PGT to families lacking standard phasing reference(s). A rapid pre-PGT work-up can define whether the couple can benefit from this technology. Importantly, by including more than one extended family member, the chance of obtaining informative results in the interrogated locus increases. Furthermore, by following the familial segregation of the variant of interest using IBS information, more evidence can be obtained regarding its pathogenicity. This is important for making the decision if the specific indication is valid for PGT.