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Chronic hepatitis B virus (HBV) infection is a major cause of liver cirrhosis and liver cancer, despite strong prevention and treatment efforts. The study of the epigenetic modification of HBV has become a research hotspot, including the N6-methyladenosine (m6A) modification of HBV RNA, which plays complex roles in the HBV life cycle. In addition to m6A modification, 5-methylcytosine (m5C) is another major modification of eukaryotic mRNA. In this study, we explored the roles of m5C methyltransferase and demethyltransferase in the HBV life cycle. The results showed that m5C methyltransferase NSUN2 deficiency could negatively regulate the expression of HBV while m5C demethyltransferase TET2 deficiency positively regulates the expression of HBV. Subsequently, we combined both in vitro bisulfite sequencing and high-throughput bisulfite sequencing methods to determine the distribution and stoichiometry of m5C modification in HBV RNA. Two sites: C2017 and C131 with the highest-ranking methylation rates were identified, and mutations at these two sites could lead to the decreased expression and replication of HBV, while the mutation of the "fake" m5C site had no effect. Mechanistically, NSUN2-mediated m5C modification promotes the stability of HBV RNA. In addition, compared with wild-type HepG2-NTCP cells and primary human hepatocytes, the replication level of HBV after NSUN2 knockdown decreased, and the ability of the mutant virus to infect and replicate in wild-type HepG2-NTCP cells and PHHs was substantially impaired. Similar results were found in the experiments using C57BL/6JGpt-Nsun2+/- mice. Interestingly, we also found that HBV expression and core protein promoted the endogenous expression of NSUN2, which implied a positive feedback loop. In summary, our study provides an accurate and high-resolution m5C profile of HBV RNA and reveals that NSUN2-mediated m5C modification of HBV RNA positively regulates HBV replication by maintaining RNA stability. Author summary: The study of the epigenetic modification of viral RNA has become a research hotspot, including the N6-methyladenosine (m6A) modification and 5-methylcytosine (m5C). However, compared with m6A modification, fewer studies on RNA m5C modification regulating viruses have been reported. This is partly due to the low abundance of m5C modification in mRNA, and the limits of sequencing technology, which hinder the identification of accurate and repeatable m5C landscape of virus RNA. Here, we discovered that depletion of NSUN2, an m5C methyltransferase, could negatively regulate the replication of hepatitis B virus (HBV). Importantly, we combined in vitro bisulfite sequencing and high-throughput bisulfite sequencing methods to determine the distribution of HBV RNA m5C modification. Two sites modified by NSUN2 with the highest-ranking methylation rates were identified, and mutations of these two sites could lead to decreased replication of HBV. Mechanistically, m5C modification promotes the stability of HBV RNA. Taken together, we provide a high-resolution and accurate atlas of m5C in HBV RNA and reveal a functional role of m5C in viral replication, which provides potential targets for the development of antiviral drugs. [ABSTRACT FROM AUTHOR]

The facing is a key component for the design of soil nail walls. However, current design specifications of soil nail walls in China do not specify methods for estimation of facing tensile forces, which has posed challenges for engineers to produce consistent, safe, and cost-effective facing designs. This study first constructs the facing tensile force as the product of the maximum nail load and a facing factor. Then a database containing 91 measured facing tensile force data collected from the literature is developed and used to calibrate the facing factor matching six maximum nail load models that are commonly adopted in China, namely, the simplified incremental calculation models based on the Rankine and Coulomb earth pressure theories, the China Academy of Building Research model, and the China Association for Engineering Construction Standardization model. Based on the developed database, the accuracies of the six proposed models for facing tensile forces are demonstrated to be unbiased on average and the dispersions in the prediction accuracy are ranked from low to medium. Furthermore, the influences of three wall working conditions on prediction accuracy of the developed models are investigated and shown to be practically insignificant. Last, the model biases are characterised as lognormal random variables. [ABSTRACT FROM AUTHOR]

Municipal water mains are built with a target service age of several decades. In such a long life, breaks can occur, even multiple times. Water mains can be maintained before or right at breaks. The former is referred to as the preventive strategy, whereas the latter is the corrective strategy. Depending on the costs of repair, replacement, and failure consequence, different strategies should typically be implemented in order to achieve the optimal watermain management in terms of life cycle costs. This study aims to investigate the optimal scenarios for the two strategies based on a two-time-scale (TTS) point process used to model the deterioration of water mains. The corrective strategy is to determine the optimal number n , where upon the n -th break, implementing a replacement for water main is justified, compared to a minimal repair. The preventive strategy is to determine the optimal replacement time in terms of pipe survival probability P s . Monte Carlo simulations are used to investigate the optimal n and P s considering a number of influential factors, including model parameters of the intensity function and ratios of maintenance, replacement, and consequence costs. Then, the full distributions of the life cycle costs are characterized with the mean of total life cycle costs being the target for optimization. Last, a case study is illustrated to demonstrate the application of both strategies in real water systems. An important finding is that with a typical pipe diameter of 400 mm and length of 200 m, the optimal n is typically less than five, and the optimal P s is below 50%. [ABSTRACT FROM AUTHOR]

Aims: Osteocalcin contributes to the regulation of endocrine system. However, the association between osteocalcin and ketosis has not been evaluated. We thus aimed to explore the relationship between total osteocalcin and risk of ketosis in type 2 diabetes (T2DM).Materials and Methods: We identified 6157 diabetes patients from Shanghai Tenth People's Hospital between 1 January 2011 and 1 March 2017. Six hundred eight subjects were enrolled in the retrospective cross-sectional study: 304 T2DM patients with ketosis whose age, gender, and body mass index were matched with 304 T2DM patients without ketosis. A further retrospective nested case-control study was conducted in 252 T2DM patients without ketosis for a mean duration of 21.58 ± 12.43 months to investigate the occurrence of ketosis.Results: Osteocalcin levels were negatively correlated with blood ketones (adjusted r = -0.263) and urine ketones (adjusted r = -0.183). The inverse dose-dependent relationship of osteocalcin and risk of ketosis was present across osteocalcin level quintiles (top quintile as the reference, adjusted odds ratio [95% CI] = 2.56 [0.80-8.17], 3.71 [0.90-15.29], 10.77 [2.63-44.15], 23.81 [4.32-131.17] per osteocalcin quintile, respectively). Ketosis occurred in 17 of the 252 T2DM patients during follow-up. The Cox regression analysis indicated that osteocalcin was an independent protective factor against development of ketosis (adjusted hazard ratio [95% CI]: 0.668 [0.460-0.971]).Conclusions: Total osteocalcin can be used as a predictor of ketosis in T2DM. [ABSTRACT FROM AUTHOR]

Aims/Introduction To investigate whether donor liver steatosis increases the incidence of new-onset diabetes after transplantation ( NODAT) in liver transplant recipients. Materials and Methods We retrospectively analyzed liver transplant recipients at Zhongshan Hospital, Shanghai, China, from April 2001 to December 2014. The final analysis involved 763 patients. The cumulative incidence of NODAT at 1, 3, 5 and 10 years after liver transplantation was investigated. Furthermore, according to the findings of donor liver biopsy before transplantation, patients were divided into steatotic and non-steatotic donor liver groups, and NODAT incidence was compared between these groups. Multivariate Cox regression was used to explore the risk factors for NODAT in the patients. Results Of the 763 donors, 309 (40.5%) had liver steatosis. At the end of follow up, 130 (42.1%) patients in the steatotic donor liver group developed NODAT, an incidence that exceeded that in the non-steatotic donor liver group ( P = 0.001). The cumulative incidence of NODAT among all patients at 1, 3, 5, and 10 years after transplantation was 33, 43, 50 and 56%, respectively. The cumulative incidences of NODAT at 1, 3, 5 and 10 years in the steatotic donor liver group were significantly higher than those in the non-steatotic donor liver group ( P = 0.003). Multivariate Cox regression analyses showed that donor liver steatosis was an independent risk factor for NODAT among liver transplant recipients, after other potential risk factors were adjusted for (hazard ratio 1.774, 95% confidence interval: 1.025-3.073; P = 0.041). Conclusions Donor liver steatosis increases NODAT incidence among liver transplant recipients. [ABSTRACT FROM AUTHOR]

Using particle agglomeration to help separate fine particles receives increasing attention in air purification processes. Recent studies suggested that enhanced turbulence using bluff bodies can significantly improve the agglomeration of fine particles. However, since micron-sized particles are difficult to measure, the current exploration of flow fields and particles is mainly based on simulations and lacks experimental validation. This paper innovatively used a phase doppler particle analyzer to measure the flow field in terms of velocity, streamline and turbulence intensity, as well as particle size and number concentration in an agglomeration chamber with a bluff body located at the center. Different shaped bluff bodies were also considered. It was found that low-velocity recirculation region with different shapes appeared in the wake, and vortex flow with special structures were observed. From the perspective of particles, the development of the flow field generated by a single bluff body effectively caused an increase in particle size and a decrease in particle number, which is because that the flow field promotes collisional agglomeration. More specifically, for V-structure bluff body, the particle size improved from 6 μm to a maximum of 13 μm and the particle number concentration decreased by 8 times. The agglomeration efficiency for PM 2.5 and PM 10 reached 89.06% and 87.38% respectively. Furthermore, by analyzing the velocity of different sized particles, it was reasonable to assume that different sized particles appeared to have velocity differences due to recirculating vortex flow, while the overlapping trajectories of positive and negative moving particles was mainly responsible for the collision probability enhancement. This paper presents new insights into particle agglomeration separation for this environment-friendly and low-cost technology. [Display omitted] • Flow field and particles were observed simultaneously by PDPA experiments. • The addition of the bluff body considerably enhances the particle size. • V-structure has the most considerable effect in enhancing the particle size. • Different sized particles show significant velocity differences in the wake. • Particle-Particle collision probability is improved by recirculating vortices. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Diabetes is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Diabetes is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Highlights: [•] Curcumol suppresses osteoclasts differentiation in vitro. [•] Curcumol impairs JNK/AP-1 signaling pathway. [•] Curcumol may be used for treating osteoclast related diseases. [ABSTRACT FROM AUTHOR]

Animal models of COVID-19 facilitate the development of vaccines and antivirals against SARS-CoV-2. The efficacy of antivirals or vaccines may differ in different animal models with varied degrees of disease. Here, we introduce a mouse model expressing human angiotensin-converting enzyme 2 (ACE2). In this model, ACE2 with the human cytokeratin 18 promoter was knocked into the Hipp11 locus of C57BL/6J mouse by CRISPR - Cas9 (K18-hACE2 KI). Upon intranasal inoculation with high (3 × 10 5 PFU) or low (2.5 × 10 2 PFU) dose of SARS-CoV-2 wildtype (WT), Delta, Omicron BA.1, or Omicron BA.2 variants, all mice showed obvious infection symptoms, including weight loss, high viral loads in the lung, and interstitial pneumonia. 100% lethality was observed in K18-hACE2 KI mice infected by variants with a delay of endpoint for Delta and BA.1, and a significantly attenuated pathogenicity was observed for BA.2. The pneumonia of infected mice was accompanied by the infiltration of neutrophils and pulmonary fibrosis in the lung. Compared with K18-hACE2 Tg mice and HFH4-hACE2 Tg mice, K18-hACE2 KI mice are more susceptible to SARS-CoV-2. In the antivirals test, REGN10933 and Remdesivir had limited antiviral efficacies in K18-hACE2 KI mice upon the challenge of SARS-CoV-2 infections, while Nirmatrelvir, monoclonal antibody 4G4, and mRNA vaccines potently protected the mice from death. Our results suggest that the K18-hACE2 KI mouse model is lethal and stable for SARS-CoV-2 infection, and is practicable and stringent to antiviral development.

Chronic hepatitis B virus (HBV) infection is a major cause of liver cirrhosis and liver cancer, despite strong prevention and treatment efforts. The study of the epigenetic modification of HBV has become a research hotspot, including the N6-methyladenosine (m6A) modification of HBV RNA, which plays complex roles in the HBV life cycle. In addition to m6A modification, 5-methylcytosine (m5C) is another major modification of eukaryotic mRNA. In this study, we explored the roles of m5C methyltransferase and demethyltransferase in the HBV life cycle. The results showed that m5C methyltransferase NSUN2 deficiency could negatively regulate the expression of HBV while m5C demethyltransferase TET2 deficiency positively regulates the expression of HBV. Subsequently, we combined both in vitro bisulfite sequencing and high-throughput bisulfite sequencing methods to determine the distribution and stoichiometry of m5C modification in HBV RNA. Two sites: C2017 and C131 with the highest-ranking methylation rates were identified, and mutations at these two sites could lead to the decreased expression and replication of HBV, while the mutation of the "fake" m5C site had no effect. Mechanistically, NSUN2-mediated m5C modification promotes the stability of HBV RNA. In addition, compared with wild-type HepG2-NTCP cells and primary human hepatocytes, the replication level of HBV after NSUN2 knockdown decreased, and the ability of the mutant virus to infect and replicate in wild-type HepG2-NTCP cells and PHHs was substantially impaired. Similar results were found in the experiments using C57BL/6JGpt-Nsun2+/- mice. Interestingly, we also found that HBV expression and core protein promoted the endogenous expression of NSUN2, which implied a positive feedback loop. In summary, our study provides an accurate and high-resolution m5C profile of HBV RNA and reveals that NSUN2-mediated m5C modification of HBV RNA positively regulates HBV replication by maintaining RNA stability., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Feng et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)