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Xiao-Xi Lin,1,* Yu-Qi Yin,2,* Wen-Zheng Zhang,3 Jia-Huan Huang,4 Wei-Shang Hu1 1Geriatric Medicine Center, Department No. 2 of Acupuncture & Massage, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, Hangzhou, Zhejiang, 310024, People’s Republic of China; 2The Third Clinical Medical College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310053, People’s Republic of China; 3Emergency and Critical Care Center, Department of Intensive Care Unit, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, Hangzhou, Zhejiang, 310024, People’s Republic of China; 4Geriatric Medicine Center, Department of General Practice, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, Hangzhou, Zhejiang, 310024, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wei-Shang Hu, Geriatric Medicine Center, Department No. 2 of Acupuncture & Massage, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital of Hangzhou Medical College, No. 1 of WangJiangShan Road, XiHu District, Hangzhou, Zhejiang, 310000, People’s Republic of China, Tel +86 571 87091302, Fax +86 571 87091089, Email huweishanghwsh@126.comBackground: During the Omicron variant outbreak of COVID-19 (2022– 2023), Chinese healthcare institutions combined traditional Chinese medicine (TCM) with Western medical practices to treat COVID-19 patients, especially the elderly. The efficacy and safety of this approach, especially for individuals aged over 85, need further investigation.Methods: In this retrospective study, a cohort of 62 patients aged over 85 years, diagnosed with COVID-19 infection, was examined. Among them, 34 patients were administered Shashen-Maidong decoction in conjunction with Western medicine (SMD+WM group), while the remaining 28 patients received only Western medicine (WM group). Comparative analysis was conducted between the two groups, encompassing parameters such as the duration for the nucleic acid test to turn negative, length of intensive care unit (ICU) stay, mortality rate, utilization of high-flow nasal cannula oxygen (HFNC), occurrence of endo-tracheal intubation, frequency of recurrent respiratory infections within three months, and various laboratory indicators.Results: There were no significant differences observed between the two groups in terms of the duration for the nucleic acid test to turn negative, the length of ICU stay, mortality rate, utilization of HFNC, performance of endo-tracheal intubation, or the frequency of recurrent respiratory infections within three months (P > 0.05). However, in comparison to the WM group, the SMD+WM group exhibited notably lower growth rates in white blood cell (WBC) and neutrophil (NEUT) values. Additionally, the SMD+WM group demonstrated superior improvement in cardiac troponin I (cTnI) and B-type natriuretic peptide (BNP) values.Conclusion: In contrast to the administration of Western medicine alone, the combined use of Shashen-Maidong decoction with Western medicine significantly suppresses the increase in WBC count, particularly in NEUT levels, in elderly patients diagnosed with COVID-19. Moreover, this combined treatment exhibits a protective effect on cardiac function and demonstrates a relatively safe profile.Keywords: COVID-19, traditional Chinese medicine, Shashen-Maidong decoction

Mammalian cells are commonly used as hosts in cell culture for biologics production in the pharmaceutical industry. Structured mechanistic models of metabolism have been used to capture complex cellular mechanisms that contribute to varying metabolic shifts in different cell lines. However, little research has focused on the impact of temporal changes in enzyme abundance and activity on the modeling of cell metabolism. In this work, we present a framework for constructing mechanistic models of metabolism that integrate growth-signaling control of enzyme activity and transcript dynamics. The proposed approach is applied to build models for three Chinese hamster ovary (CHO) cell lines using fed-batch culture data and time-series transcript profiles. Leveraging information from the transcriptome data, we develop a parameter estimation approach based on multi-cell-line (MCL) learning, which combines data sets from different cell lines and trains the individual cell-line models jointly to improve model accuracy. The computational results demonstrate the important role of growth signaling and transcript variability in metabolic models as well as the virtue of the MCL approach for constructing cell-line models with a limited amount of data. The resulting models exhibit a high level of accuracy in predicting distinct metabolic behaviors in the different cell lines; these models can potentially be used to accelerate the process and cell-line development for the biomanufacturing of new protein therapeutics., (© 2024 The Author(s). Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

To explore the incidence of new-onset erectile dysfunction (ED) in diabetes mellitus (DM) patients with sodium-glucose cotransporter 2 inhibitors (SGLT2I) use compared to a control group of non-SGLT2I use by propensity matching (PS) matching approach. Cox proportion hazards regression models were used to examine the effect of SGLT2I and risk factors on the risk of developing ED, presented as a hazard ratio (HR) with a 95% confidence interval (CI). 159773 DM patients using SGLT2I and 159773 PS-matching DM patients who had never used SGLT2I was included. SGLT2I users had a higher risk of ED than the non-SGLT2I users (adjusted HR = 1.55, 95% CI = 1.40-1.72). The likelihood of developing ED was higher in patients with SGLT2I use was found., Competing Interests: The authors declare no conflicts of interest, (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

HIV-1, the causative agent of AIDS, is a retrovirus that packages two copies of unspliced viral RNA as a dimer into newly budding virions. The unspliced viral RNA also serves as an mRNA template for translation of two polyproteins. Recent studies suggest that the fate of the viral RNA (genome or mRNA) is determined at the level of transcription. RNA polymerase II uses heterogeneous transcription start sites to generate major transcripts that differ in only two guanosines at the 5' end. Remarkably, this two-nucleotide difference is sufficient to alter the structure of the 5'-untranslated region and generate two pools of RNA with distinct functions. The presence of both RNA species is needed for optimal viral replication and fitness., Competing Interests: Declaration of competing interest None., (Published by Elsevier Ltd.)

The objective was to assess the protective role of anti-diabetic agent (ADA) in predicting interstitial lung disease (ILD) among patients with diabetes mellitus (DM). We formed a cohort of DM patients between 2009 and 2016 using data from Taiwan. Univariable and multivariable Cox proportion hazards regression models were used to examine the effect of risk factor on the risk of developing ILD, presented as a hazard ratio (HR) with a 95% confidence interval (CI). Cox proportional hazard regression analysis for the risk of DM-associated ILD with joint effect of dipeptidyl peptidase-4 inhibitor (DPP4I), glucagon-like peptide-1 receptor agonist (GLP-1 RA), and sodium-glucose cotransporter 2 inhibitors (SGLT2I) showed that SGLT2I, GLP-1 RA, and DPP-4I had a decreased risk of ILD with adjusted HR of 0.14 (0.11, 0.18), 0.29 (0.24, 0.35), and 0.64 (0.62, 0.67), respectively. DPP4I, GLP-1 RA, and SGLT2I could be considered to be introduced to this DM population for ILD risk reduction in DM, especially with SGLT2I usage., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Recombinant adeno-associated virus (rAAV) is a major gene delivery vehicle. We have constructed a stable rAAV producer cell line by integrating essential rAAV genome, viral and helper genes into the genome of HEK293 cell under the control of inducible promoters. Upon induction, the cell line produces transducing rAAV. To gain insight into enhancing rAAV productivity and vector quality, we performed a comparative transcriptomic and proteomic analysis of our synthetic cell line GX2 and two wild-type AAV (wtAAV) production systems, one by virus co-infection and the other by multi-plasmid transfection. The three systems had different kinetics in viral component synthesis but generated comparable copies of AAV genomes; however, the capsid titer of GX2 was an order of magnitude lower compared to those two wtAAV systems, indicating that its capsid production may be insufficient. The genome packaging efficiency was also lower in GX2 despite it produced higher levels of Rep52 proteins than either wtAAV systems, suggesting that Rep52 protein expression may not limit genome packaging. In the two wtAAV systems, VP were the most abundant AAV proteins and their levels continued to increase, while GX2 had high level of wasteful cargo gene expression. Furthermore, upregulated inflammation, innate immune responses, and MAPK signaling, as well as downregulated mitochondrial functions, were commonly observed in either rAAV or wtAAV systems. Overall, this comparative multi-omics study provided rich insights into host cell and viral factors that are potential targets for genetic and process intervention to enhance the productivity of synthetic rAAV producer cell lines. KEY POINTS: • wtAAV infection was more efficient in producing full viral particles than the synthetic cell GX2. • Capsid protein synthesis, genome replication, and packaging may limit rAAV production in GX2. • wtAAV infection and rAAV production in GX2 elicited similar host cell responses., (© 2024. The Author(s).)

Multisource remote sensing data classification is a challenging research topic, and how to address the inherent heterogeneity between multimodal data while exploring their complementarity is crucial. Existing deep learning models usually directly adopt feature-level fusion designs, most of which, however, fail to overcome the impact of heterogeneity, limiting their performance. As such, a multimodal joint classification framework, called global clue-guided cross-memory quaternion transformer network (GCCQTNet), is proposed for multisource data i.e., hyperspectral image (HSI) and synthetic aperture radar (SAR)/light detection and ranging (LiDAR) classification. First, a three-branch structure is built to extract the local and global features, where an independent squeeze-expansion-like fusion (ISEF) structure is designed to update the local and global representations by considering the global information as an agent, suppressing the negative impact of multimodal heterogeneity layer by layer. A cross-memory quaternion transformer (CMQT) structure is further constructed to model the complex inner relationships between the intramodality and intermodality features to capture more discriminative fusion features that fully characterize multimodal complementarity. Finally, a cross-modality comparative learning (CMCL) structure is developed to impose the consistency constraint on global information learning, which, in conjunction with a classification head, is used to guide the end-to-end training of GCCQTNet. Extensive experiments on three public multisource remote sensing datasets illustrate the superiority of our GCCQTNet with regards to other state-of-the-art methods.

Scope: Little is known about the effect of blood vitamin D status on the gut mycobiota (i.e., fungi), a crucial component of the gut microbial ecosystem. The study aims to explore the association between 25-hydroxyvitamin D [25(OH)D] and gut mycobiota and to investigate the link between the identified mycobial features and blood glycemic traits., Methods and Results: The study examines the association between serum 25(OH)D levels and the gut mycobiota in the Westlake Precision Birth Cohort, which includes pregnant women with gestational diabetes mellitus (GDM). The study develops a genetic risk score (GRS) for 25(OH)D to validate the observational results. In both the prospective and cross-sectional analyses, the vitamin D is associated with gut mycobiota diversity. Specifically, the abundance of Saccharomyces is significantly lower in the vitamin D-sufficient group than in the vitamin D-deficient group. The GRS of 25(OH)D is inversely associated with the abundance of Saccharomyces. Moreover, the Saccharomyces is positively associated with blood glucose levels., Conclusion: Blood vitamin D status is associated with the diversity and composition of gut mycobiota in women with GDM, which may provide new insights into the mechanistic understanding of the relationship between vitamin D levels and metabolic health., (© 2024 The Authors. Molecular Nutrition & Food Research published by Wiley‐VCH GmbH.)

HIV-1 cores, which contain the viral genome and replication machinery, must disassemble (uncoat) during viral replication. However, the viral and host factors that trigger uncoating remain unidentified. Recent studies show that infectious cores enter the nucleus and uncoat near the site of integration. Here, we show that efficient uncoating of nuclear cores requires synthesis of a double-stranded DNA (dsDNA) genome >3.5 kb and that the efficiency of uncoating correlates with genome size. Core disruption by capsid inhibitors releases viral DNA, some of which integrates. However, most of the viral DNA is degraded, indicating that the intact core safeguards viral DNA. Atomic force microscopy and core content estimation reveal that synthesis of full-length genomic dsDNA induces substantial internal strain on the core to promote uncoating. We conclude that HIV-1 cores protect viral DNA from degradation by host factors and that synthesis of long double-stranded reverse transcription products is required to trigger efficient HIV-1 uncoating.

Recombinant adeno-associated virus (rAAV) is widely used as an in vivo delivery vector for gene therapy. It is used in a very large dose, and the large quantities required for broad applications present manufacturing challenges. We have developed a synthetic biology platform of constructing cell lines integrated with essential viral genes which can be induced to produce rAAV without plasmid transfection or virus transduction. Through iterative design-construct-characterization cycles, we have showcased the potential of this synthetic cell production system. Systems characterization of the dynamics of viral transcripts and proteins as well as virus assembly and packaging revealed that the expression level and balance of viral genome and capsid protein are keys to not only the productivity but also the full particle content, an important product quality attribute. Boosting cap gene expression by sequential transfection and integration of multiple copies of the cap gene elevated the rAAV titer to levels on a par with traditional plasmid transfection and virus infection. However, overexpression of the cap gene shifted the balance and kinetics of the genome and capsid. We independently tuned the dynamics of genome amplification and capsid protein synthesis by modulating the induction concentration as well as the time profile, and significantly enhanced full particle content while maintaining a high productivity. This strategy of constructing an inducible stable producer cell line is readily adaptable to rAAV vectors of different serotypes and payloads. It can greatly facilitate scalable production of gene therapy vectors., (© 2024 The Authors. Biotechnology Journal published by Wiley-VCH GmbH.)

Recombinant adeno-associated virus (rAAV) is among the most commonly used vectors for gene therapy. It is commonly produced by transfection of HEK293 cells with three plasmids each containing the vector genome including gene of interest (GOI), helper functions, and rep and cap genes for genome replication and capsid formation. To meet the potential clinical needs, the productivity of the production system needs to be enhanced. A better process characterization of the production system will further advance our insights into ways to enhance productivity. Here, we employed transcriptomic analysis to quantify the dynamics of different isoforms of viral transcripts and to assess the shift of cellular physiology, and deployed targeted proteomic analysis for absolute quantification of viral proteins and tandem mass tags (TMTs) for assessing cellular responses at the protein level. Functional analysis at transcriptome and proteome levels identified defense and immune response, unfolded protein response, p53 signaling as enriched. The small molecule additive intervention study based on functional analysis showed the potential of such omics-guided productivity enhancement. Together, multi-omics analysis advanced understanding of rAAV production and provided insight into enhancing rAAV production by plasmid transfection., (© 2024 The Authors. Biotechnology Progress published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)

This research is an attempt to investigate the benefit of sodium-glucose cotransporter-2 inhibitor (SGLT2I) use in patients with diabetes mellitus (DM) for outcomes of sepsis/septic shock. We used Taiwan's national data set to identify patients and patients' characteristics to investigate sepsis/septic shock among diabetes patients who use SGLT2I compared to those who do not. We have compared the two groups for several relevant categories of potential risk factors for sepsis/septic shock and adjusted the Cox regression models accordingly. The adapted diabetes complications severity index (DCSI) was used for stratifying the advancing disease of DM. Compared to patients with DCSI = 0, patients with DCSI ≥ 2 had a significantly higher risk of sepsis/septic shock (adjusted HR = 1.52, 95% CI = 1.37-1.68). A significantly lower risk of sepsis/septic shock events was observed in the SGLT2I cohort than in the non-SGLT2I cohort with the DCSI groups [adjusted HR = 0.6 (DCSI group = 0), adjusted HR = 0.61 (DCSI group = 1), adjusted HR = 0.55 (DCSI group ≥ 2)]. Patients who received SGLT2I for a cumulative duration of ≥ 90 days had a significantly lower risk of sepsis/septic shock than patients with a duration of < 90 days (adjusted HR = 0.36, 95% CI = 0.34-0.39). We described a decreased risk of sepsis/septic shock among diabetic patients who took SGLT2I., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Influenza A virus (IAV) is a negative-sense RNA virus that causes seasonal infections and periodic pandemics, inflicting huge economic and human costs on society. The current production of influenza virus for vaccines is initiated by generating a seed virus through the transfection of multiple plasmids in HEK293 cells followed by the infection of seed viruses into embryonated chicken eggs or cultured mammalian cells. We took a system design and synthetic biology approach to engineer cell lines that can be induced to produce all viral components except hemagglutinin (HA) and neuraminidase (NA), which are the antigens that specify the variants of IAV. Upon the transfection of HA and NA, the cell line can produce infectious IAV particles. RNA-Seq transcriptome analysis revealed inefficient synthesis of viral RNA and upregulated expression of genes involved in host response to viral infection as potential limiting factors and offered possible targets for enhancing the productivity of the synthetic cell line. Overall, we showed for the first time that it was possible to create packaging cell lines for the production of a cytopathic negative-sense RNA virus. The approach allows for the exploitation of altered kinetics of the synthesis of viral components and offers a new method for manufacturing viral vaccines.

Objective: To investigate predictors of anticoagulation efficacy in deep venous thrombosis (DVT) by ultrasound elastography (UE)., Methods: The basic clinical, laboratory and ultrasound treatment data of fifty-eight patients with DVT were collected and analyzed. Then the results of ultrasound after 3-month anticoagulation treatment were compared among different groups. Multiple logistic regression analysis was used to identify independent risk factors that affected anticoagulation efficacy. The predictive efficacy of each independent risk factor was accessed by drawing operating characteristic (ROC) curves., Results: According to the regression analysis, the elastic modulus (OR = 0.631, P = 0.001) and strain rate ratio (OR = 0.332, P = 0.006) were identified as independent risk factors for the effectiveness of anticoagulation therapy in patients with DVT. According to the ROC curves, elastic modulus and strain rate ratio could predict effective anticoagulation therapy for DVT, and the optimal threshold values were 22.10 kPa and 1.80 respectively. The corresponding AUC values were 0.879 and 0.854, with a sensitivity of 71.4% and 59.5%, a specificity of 93.7%, and a Youden index of 65.1% and 62.7%, respectively., Conclusions: The elastic modulus (≤22.10 kPa) or strain rate ratio (≤1.80) of the thrombus were independent predictors for the effectiveness of anticoagulation therapy.

Live-cell imaging has become a powerful tool for dissecting the behavior of viral complexes during HIV-1 infection with high temporal and spatial resolution. Very few HIV-1 particles in a viral population are infectious and successfully complete replication (~1/50). Single-particle live-cell imaging enables the study of these rare infectious viral particles, which cannot be accomplished in biochemical assays that measure the average property of the entire viral population, most of which are not infectious. The timing and location of many events in the early stage of the HIV-1 life cycle, including nuclear import, uncoating, and integration, have only recently been elucidated. Live-cell imaging also provides a valuable approach to study interactions of viral and host factors in distinct cellular compartments and at specific stages of viral replication. Successful live-cell imaging experiments require careful consideration of the fluorescent labeling method used and avoid or minimize its potential impact on normal viral replication and produce misleading results. Ideally, it is beneficial to utilize multiple virus labeling strategies and compare the results to ensure that the virion labeling did not adversely influence the viral replication step that is under investigation. Another potential benefit of using different labeling strategies is that they can provide information about the state of the viral complexes. Here, we describe our methods that utilize multiple fluorescent protein labeling approaches to visualize and quantify important events in the HIV-1 life cycle, including docking HIV-1 particles with the nuclear envelope (NE) and their nuclear import, uncoating, and proviral transcription., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Recombinant adeno-associated virus (rAAV) is among the most commonly used in vivo gene delivery vehicles and has seen a number of successes in clinical application. Current manufacturing processes of rAAV employ multiple plasmid transfection or rely on virus infection and face challenges in scale-up. A synthetic biology approach was taken to generate stable cell lines with integrated genetic modules, which produced rAAV upon induction albeit at a low productivity. To identify potential factors that restrained the productivity, we systematically characterized virus production kinetics through targeted quantitative proteomics and various physical assays of viral components. We demonstrated that reducing the excessive expression of gene of interest by its conditional expression greatly increased the productivity of these synthetic cell lines. Further enhancement was gained by optimizing induction profiles and alleviating proteasomal degradation of viral capsid protein by the addition of proteasome inhibitors. Altogether, these enhancements brought the productivity close to traditional multiple plasmid transfection. The rAAV produced had comparable full particle contents as those produced by conventional transient plasmid transfection. The present work exemplified the versatility of our synthetic biology-based viral vector production platform and its potential for plasmid- and virus-free rAAV manufacturing., (© 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)

HIV-1 virions incorporate viral RNA, cellular RNAs, and proteins during the assembly process. Some of these components, such as the viral RNA genome and viral proteins, are essential for viral replication, whereas others, such as host innate immune proteins, can inhibit virus replication. Therefore, analyzing the virion content is an integral part of studying HIV-1 replication. Traditionally, virion contents have been examined using biochemical assays, which can provide information on the presence or absence of the molecule of interest but not its distribution in the virion population. Here, we describe a method, single-virion analysis, that directly examines the presence of molecules of interest in individual viral particles using fluorescence microscopy. Thus, this method can detect both the presence and the distribution of molecules of interest in the virion population. Single-virion analysis was first developed to study HIV-1 RNA genome packaging. In this assay, HIV-1 unspliced RNA is labeled with a fluorescently tagged RNA-binding protein (protein A) and some of the Gag proteins are labeled with a different fluorescent protein (protein B). Using fluorescence microscopy, HIV-1 particles can be identified by the fluorescent protein B signal and the presence of unspliced HIV-1 RNA can be identified by the fluorescent protein A signal. Therefore, the proportions of particles that contain unspliced RNA can be determined by the fraction of Gag particles that also have a colocalized RNA signal. By tagging the molecule of interest with fluorescent proteins, single-virion analysis can be easily adapted to study the incorporation of other viral or host cell molecules into particles. Indeed, this method has been adapted to examine the proportion of HIV-1 particles that contain APOBEC3 proteins and the fraction of particles that contain a modified Gag protein. Therefore, single-virion analysis is a flexible method to study the nucleic acid and protein content of HIV-1 particles., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

To compare the potential role of sodium-glucose cotransporter-2 inhibitors (SGLT2I) in the development of psychiatric disease among patients with type 2 diabetes mellitus (DM). Using a large population-based database, SGLT2I users and non-SGLT2I users were 1:1 matched according to the covariates of sex, age, comorbidities, adapted diabetes complications severity index (DCSI), medications, and index year using propensity score matching and a logistic regression model. We calculated the incidence of major psychiatric disorders and adjusted hazard ratios (HR) with 95% confidence interval (CI) for SGLT2I users and the non- SGLT2I users using a Cox proportional hazards model. SGLT2I were associated with a lower risk for psychiatric disorders than those not treated with SGLT2I (HR 0.80 and 95% CI 0.72-0.88). Among patients with DM, SGLT2I were associated with a lower risk of psychiatric disease., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

In July 2022, the World Health Organization announced monkeypox as a public health emergency of international concern (PHEIC), and over 85,000 global cases have been reported currently. However, preventive and therapeutic treatments for the monkeypox virus (MPXV) remain limited. MPXV mRNA cap N7 methyltransferase (MTase) is composed of two subunits (E1 C-terminal domain (E1 CTD ) and E12) which are essential for the replication of MPXV. Here, we solved a 2.16 Å crystal structure of E12. We also docked the D1 CTD of the vaccinia virus (VACV) corresponding to the E1 CTD in MPXV with E12 and found critical residues at their interface. These residues were further used for drug screening. After virtual screening, the top 347 compounds were screened out and a list of top 20 potential MPXV E12 inhibitors were discovered, including Rutin, Quercitrin, Epigallocatechin, Rosuvastatin, 5-hydroxy-L-Tryptophan, and Deferasirox, etc., which were potential E12 inhibitors. Taking the advantage of the previously unrecognized special structure of MPXV MTase composing of E1 CTD and E12 heterodimer, we screened for inhibitors targeting MTase for the first time based on the interface between the heterodimer of MPXV MTase. Our study may provide insights into the development of anti-MPXV drugs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Radial artery cannulation helps to maintain the stability of maternal hemodynamics and reduce complications; however, it is difficult for women with gestational hypertension. Subcutaneous nitroglycerin was found to improve the first attempt success rate of radial artery cannulation in pediatric patients. Therefore, this study evaluated the effect of subcutaneous nitroglycerin on the radial artery diameter and area, blood flow rate and the success rate of radial artery cannulation in women with pregnancy-induced hypertension., Methods: A total of 94 women with gestational hypertension and risk of intraoperative bleeding undergoing cesarean section were identified and randomized into the subcutaneous nitroglycerin group and control group. The primary outcome was the success rate of left radial artery cannulation within 3 min after subcutaneous injecting (T2). The puncture time, number of attempts, the overall complications, and ultrasonographic measurements including radial artery diameter, cross-sectional area and depth were also recorded before subcutaneous injection (T1), 3 min after subcutaneous injection (T2) and immediately after radial artery cannulation (T3)., Results: The first attempt success rate of radial artery cannulation was significantly higher (97.9% vs. 76.6%, p = 0.004) and procedure time to success was significantly shorter (111 ± 18 s vs. 171 ± 70 s, p < 0.001) in the subcutaneous nitroglycerin group as compared to the control group. The subcutaneous nitroglycerin group also had a significantly less overall number of attempts as 1/2/3 attempts (n), 46/1/0 vs. 36/7/4 (p = 0.008). Compared with the control group, the diameter and cross-sectional area of radial artery increased significantly at the T2 and T3 points in the subcutaneous nitroglycerin group (p < 0.001), as well as percentage change of radial artery diameter and CSA. Vasospasm (6.4% vs. 31.9%; p = 0.003) was significantly lower in the subcutaneous nitroglycerin group; however, no difference was found in hematoma (2.1% vs. 12.8%; p = 0.111)., Conclusion: Subcutaneous nitroglycerin along with the routine local anesthetic preparation before radial artery cannulation increased the first attempt success rate of radial artery cannulation and decreased the overall number of cannulation attempts in women with gestational hypertension and risks of intraoperative bleeding undergoing cesarean section, it also decreased cannulation times and overall number of vasospasms., (© 2023. The Author(s).)

Background: Aging is a biological and gradual deterioration of function in living organisms. Aging is one of the risk factors for heart disease., Objective: Although mesenchymal stem cell transplantation shows potential in heart disease treatment, the relationship between stem cell-based therapy and oxidative stress/inflammasome axis regulation remains unclear. This study hypothesized that intervention of stem cells showed a protective effect on heart aging induced by D-galactose through regulation of oxidative stress/inflammasome axis., Methods: An aging animal model was designed to test the above hypothesis. Experimental animals were divided into three groups, including Sham, D-gal (aging rats induced by d-galactose), and D-gal+WJSC (aging rats receiving mesenchymal stem cells)., Results: Compared to the Sham, the experimental results indicate that structural alteration (HE stain and Masson's Trichrome stain), oxidative stress elevation (increase of TBARS level, expression of gp-91 and suppression of Sirt-1 as well as SOD2), increase of aging marker p53, suppression of cardiogenesis marker Troponin T, and inflammasome related protein markers expression (NLRP3, caspase-1 and IL-1 beta) were significantly observed in D-gal. In contrast, all pathological pathways were significantly improved in D-gal+WJSC when compared to D-gal. In addition, migration of stem cells to aging heart tissues was observed in the D-gal+WJSC group., Conclusion: These findings suggest that mesenchymal stem cell transplantation effectively ameliorates aging hearts through oxidative stress/inflammasome axis regulation. The results from this study provide clinical potential for stem cell-based therapy in the treatment of aging hearts., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Cell therapy offers the potential for curative treatment of cancers. Although T cells have been the predominantly used cell type, natural killer (NK) cells have attracted great attention owing to their ability to kill cancer cells and because they are naturally suitable for allogeneic applications. Upon stimulation by cytokines or activation by a target cell, NK cells proliferate and expand their population. These cytotoxic NK cells can be cryopreserved and used as an off-the-shelf medicine. The production process for NK cells thus differs from that of autologous cell therapies. We briefly outline key biological features of NK cells, review the manufacturing technologies for protein biologics, and discuss their adaptation for developing robust NK cell biomanufacturing processes., Competing Interests: Declaration of interests F.C. is a paid consultant for Fate Therapeutics and receives research funds and stock options from this relationship. He also receives research financial support from Gamida Cell. The remaining authors have no interests to declare., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

We report a retrospective analysis of a nationwide health database to study the association between sodium-glucose cotransporter-2 inhibitor (SGLT2I) use and the incidence of adverse clinical outcomes among heart failure (HF) patients with and without atrial fibrillation (AF) stratified by CHA2DS2-VASc score. The outcome of this study was on the development of adverse events, including acute myocardial infarction (AMI), hemorrhagic stroke, ischemic stroke, cardiovascular (CV) death, and all-cause mortality. By dividing the number of adverse events by the total person-years, the incidence rate was calculated. The hazard ratio (HR) was estimated by the Cox proportional hazard model. A total of 95% confidence interval (CI) was also presented to show the risk of adverse events for HF patients with and without AF taking SGLT2I. SGLT2I users had a lower risk of AMI (adjusted HR = 0.83; 95% CI = 0.74, 0.94), CV death (adjusted HR = 0.47; 95% CI = 0.42, 0.51), and all-cause death (adjusted HR = 0.39; 95% CI = 0.37, 0.41). Considering HF patients without AF and SGLT2I as the reference group, HF patients without AF but with SGLT2I had a reduced risk of adverse outcomes of 0.48 (95% CI = 0.45, 0.50), and HF patients with AF and SGLT2I had the decreased hazard ratio of 0.55 (95% CI = 0.50, 0.61). The adjusted HR of adverse outcomes for HF patients with CHA2DS2-VASc score less than 2 and SGLT2I without and with AF relative to HF patients without AF nor SGLT2I were 0.53 (95% CI = 0.41, 0.67) and 0.24 (95% CI = 0.12, 0.47), respectively. Compared to HF patients with no history of AF and SGLT2I, if patients additionally with SGLT2I and CHA2DS2-VASc score ≥ 2, the risk of the adverse outcomes was reduced with adjusted HR of 0.48 (95% CI = 0.45, 0.50); if patients additionally with AF and CHA2DS2-VASc score ≥ 2, the risk of the adverse outcomes was decreased with adjusted HR of 0.88 (95% CI = 0.80, 0.97); if patients additionally with AF, SGLT2I, and CHA2DS2-VASc score ≥ 2, the risk of the adverse outcomes was diminished with adjusted HR of 0.52 (95% CI = 0.47, 0.58). We concluded that SGLT2I has a protective effect in HF patients, and the risk reduction is greater with a score of < 2 and without AF., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Abstract: The authors report the impact of angiotensin receptor-neprilysin inhibitor (ARNI) versus renin-angiotensin system inhibitor (RASI) on the management and outcomes in dementia among heart failure (HF) patients as obtained from the real-life nationwide registry. In this study, HF patients between January 1, 2017 and December 31, 2019 were divided into 2 groups, including subjects receiving RASI and ARNI. The incidence rate of dementia was calculated with the unit of 1000 person-years. Cox proportional hazard model was applied for the examination of the hazard ratio, and also presented with 95% confidence interval. Between 2017 and 2019, RASI and ARNI cohorts contain 18,154 subjects. After adjusting with age, sex, comorbidities, and medications, ARNI cohort had a lower risk of dementia (adjusted hazard ratio = 0.83; 95% confidence interval = 0.72, 0.95) than RASI cohort. The authors concluded that use of ARNI was associated with a lower risk of new-onset dementia in patients with HF., Competing Interests: The authors report no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Gene therapy strategies that effectively inhibit HIV-1 replication are needed to reduce the requirement for lifelong antiviral therapy and potentially achieve a functional cure. We previously designed self-activating lentiviral vectors that efficiently delivered and expressed a Vif-resistant mutant of APOBEC3G (A3G-D128K) to T cells, which potently inhibited HIV-1 replication and spread with no detectable virus. Here, we developed vectors that express A3G-D128K, membrane-associated fusion inhibitor peptide mC46, and O 6 -methylguanine-DNA-methyltransferase (MGMT) selectable marker for in vivo selection of transduced CD34 + hematopoietic stem and progenitor cells. MGMT-selected T cell lines MT4, CEM, and PM1 expressing A3G-D128K (with or without mC46) potently inhibited NL4-3 infection up to 45 days post infection with no detectable viral replication. Expression of mC46 was sufficient to block infection >80% in a single-cycle assay. Importantly, expression of mC46 provided a selective advantage to the A3G-D128K-modified T cells in the presence of replication competent virus. This combinational approach to first block HIV-1 entry with mC46, and then block any breakthrough infection with A3G-D128K, could provide an effective gene therapy treatment and a potential functional cure for HIV-1 infection., Competing Interests: The authors declare no competing interests.

Abstract: This study was to evaluate the association between heart failure (HF) patients with and without sacubitril-valsartan use with incident cancer risk. This study consisted of 18,072 patients receiving sacubitril-valsartan and 18,072 control group participants. In the Fine and Gray model, which extends the standard Cox proportional hazards regression model, we estimated the relative risk of developing cancer between the sacubitril-valsartan cohort and the non-sacubitril-valsartan cohort by using subhazard ratios (SHRs) and 95% confidence intervals (CIs). The incidence rates of cancer were 12.02 per 1000 person-years for the sacubitril-valsartan cohort and 23.31 per 1000 person-years for the non-sacubitril-valsartan cohort. Patients receiving sacubitril-valsartan had a significantly lower risk of developing cancer with an adjusted SHR of 0.60 (0.51, 0.71). Sacubitril-valsartan users were less to be associated with the development of cancer., Competing Interests: The authors report no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

In the area of manipulating the spatial electromagnetic (EM) waves fields, the metasurfaces have become much more attractive and powerful in recent years. Here, we propose a design to realize the simultaneous control of spatial fundamental and harmonic EM waves applying nonlinear metasurfaces in microwave band. The proposed meta-atom composed of three topological layers which are transmitting antenna, nonlinear wave guiding and receiving antenna respectively. And the critical factor of generating the harmonic is the nonlinear chip which is integrated into the middle layer. The microstrip power divider and phase shifter in each meta-atom are preciously tailored to actualize the spatial control of the fundamental and harmonic transmission beams in the far field. One prototype of the nonlinear metasurfaces is fabricated and corresponding radiation patterns of fundamental and harmonic modes are observed very well in the experience that can verify the validity of our proposed method.

Purpose: We tried to define the association of type 2 diabetes mellitus (T2DM) patients with and without sodium-glucose cotransporter-2 inhibitors (SGLT2I) use and incident cataract using nationwide data in Taiwan., Methods: In a Cox proportional hazards regression model, we estimated the hazard ratios (HR) and 95% confidence intervals (95% CI). We considered risk factors for variables of sex, age, comorbidities and medications that we adjusted in multivariable Cox model., Results: We identified 20,768 T2DM patients in this study; 10,384 patients in the SGLT2I cohort, and 10,384 controls in the non-SGLT2I cohort. Compared with the T2DM patients without SGLT2I, T2DM patients using SGLT2I had a 2.04-fold increased risk of cataract, after adjustment by sex, age, comorbidities and medications., Conclusion: Increased risk of cataract among diabetic patients who took SGLT2I was found in this study., (© 2023. Springer-Verlag Italia S.r.l., part of Springer Nature.)

Objective: This study is on the use of the adapted Diabetes Complications Severity Index (aDCSI) for erectile dysfunction (ED) risk stratification in male patients with type 2 diabetes mellitus (DM)., Methods: This is a retrospective study with records obtained from Taiwan's National Health Insurance Research Database. Adjusted HRs (aHRs) were estimated by multivariate Cox proportional hazards models with 95% confidence intervals (CIs).., Results: A population of 84 288 eligible male patients with type 2 DM were included. Compared with change in aDCSI score of 0.0-0.5 per year, the aHRs and the corresponding 95% CIs for other changes in aDCSI scores are summarised as follows: 1.10 (0.90 to 1.34) for change in aDCSI score of 0.5-1.0 per year; 4.44 (3.47 to 5.69) for change in aDCSI score of 1.0-2.0 per year; and 10.9 (7.47 to 15.9) for change in aDCSI score of >2.0 per year.., Conclusions: Progression in aDCSI score might be used for ED risk stratification in men affected by type 2 DM., (© The Author(s) 2022. Published by Oxford University Press on behalf of Postgraduate Medical Journal. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Continuous glucose monitoring (CGM) has shown potential in improving maternal and neonatal outcomes in individuals with type 1/2 diabetes, but data in gestational diabetes mellitus (GDM) is limited. We aimed to explore the relationship between CGM-derived metrics during pregnancy and pregnancy outcomes among women with GDM., Methods: We recruited 1302 pregnant women with GDM at a mean gestational age of 26.0 weeks and followed them until delivery. Participants underwent a 14-day CGM measurement upon recruitment. The primary outcome was any adverse pregnancy outcome, defined as having at least one of the outcomes: preterm birth, large-for-gestational-age (LGA) birth, fetal distress, premature rupture of membranes, and neonatal intensive care unit (NICU) admission. The individual outcomes included in the primary outcome were considered as secondary outcomes. We conducted multivariable logistic regression to evaluate the association of CGM-derived metrics with these outcomes., Findings: Per 1-SD difference in time above range (TAR), glucose area under the curve (AUC), nighttime mean blood glucose (MBG), daytime MBG, and daily MBG was associated with higher risk of any adverse pregnancy outcome, with odds ratio: 1.22 (95% CI 1.08-1.36), 1.22 (95% CI 1.09-1.37), 1.18 (95% CI 1.05-1.32), 1.21 (95% CI 1.07-1.35), and 1.22 (95% CI 1.09-1.37), respectively. Time in range, TAR, AUC, nighttime MBG, daytime MBG, daily MBG, and mean amplitude of glucose excursions were positively associated, while time blow range was inversely associated with the risk of LGA. Additionally, higher value for TAR was associated with higher risk of NICU admission. We further summarized the potential thresholds of TAR (2.5%) and daily MBG (4.8 mmol/L) to distinguish individuals with and without any adverse pregnancy outcome., Interpretation: The CGM-derived metrics may help identify individuals at higher risk of adverse pregnancy outcomes. These CGM biomarkers could serve as potential new intervention targets to maintain a healthy pregnancy status among women with GDM., Funding: National Key R&D Program of China, National Natural Science Foundation of China, and Westlake Laboratory of Life Sciences and Biomedicine., Competing Interests: No potential conflicts of interest relevant to this article were reported., (© 2023 The Author(s).)

The objective was to assess the role of the combination approach with ezetimibe 10 mg/simvastatin 20 mg versus atorvastatin 40 mg in predicting atrial fibrillation (AF) in type 2 diabetes mellitus patients with acute coronary syndrome and acute ischemic stroke. The authors formed a cohort of diabetic patients with extensive vascular diseases between 2000 and 2018 using data from the National Health Insurance Research Database in Taiwan. AF was the outcome of interest in this study. Cox proportional hazards regression analysis was performed to estimate the hazard ratios and 95% confidence intervals in the analysis. After controlling for sex, age, comorbidities and medications, the patients coexisting with type 2 diabetes mellitus, acute coronary syndrome and acute ischemic stroke with ezetimibe 10 mg/simvastatin 20 mg treatment were not significantly at risk of AF, compared to the patients with atorvastatin 40 mg treatment (adjusted hazard ratio, 0.85; 95% confidence interval, 0.52-1.38). A similar effect for AF risk between ezetimibe 10 mg/simvastatin 20 mg and atorvastatin 40 mg users was observed in the current investigation., Competing Interests: The authors have no conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc.)

Menaquinone-7 (MK-7), a valuable member of the vitamin K2 series, is an essential nutrient for humans. It is used for treating coagulation disorders, and osteoporosis, promoting liver function recovery, and preventing cardiovascular diseases. In this study, to further improve the metabolic synthesis of MK-7 by the mutant strain, the effect of surfactants on the metabolic synthesis of MK-7 by the mutant strain Bacillus subtilis 168 KO-SinR (BS168 KO-SinR) was analyzed. The scanning electron microscopy and flow cytometry results showed that the addition of surfactants changed the permeability of the cell membrane of the mutant strain and the structural components of the biofilm. When 0.7% Tween-80 was added into the medium, the extracellular and intracellular synthesis of MK-7 reached 28.8 mg/L and 59.2 mg/L, respectively, increasing the total synthesis of MK-7 by 80.3%. Quantitative real-time PCR showed that the addition of surfactant significantly increased the expression level of MK-7 synthesis-related genes, and the electron microscopy results showed that the addition of surfactant changed the permeability of the cell membrane. The research results of this paper can serve as a reference for the industrial development of MK-7 prepared by fermentation., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Purpose: The relation between hospitalization timing and risk of clinical outcomes among patients with atrial fibrillation (AF) with and without stroke remained undetermined., Methods: Rehospitalization due to AF, cardiovascular (CV) death and all-cause mortality were the outcomes of interest in this study. Multivariable Cox proportional hazard model was applied to estimate the adjusted hazard ratio (HR) and 95% confidence interval (CI)., Results: While considering patients with AF hospitalized during weekdays without stroke as the reference group, patients with AF hospitalized during weekends with stroke had the risk of AF rehospitalization, CV death and all-cause death by 1.48 (95% CI 1.44 to 1.51), 1.77 (95% CI 1.71 to 1.83) and 1.17 (95% CI 1.15 to 1.19) times, respectively., Conclusion: Patients with AF hospitalized during weekends with stroke had the worst clinical outcomes., (© The Author(s) 2022. Published by Oxford University Press on behalf of Postgraduate Medical Journal. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

HIV-1 relies on host RNA polymeraseII (Pol II) to transcribe its genome and uses multiple transcription start sites (TSS), including three consecutive guanosines located near the U3-R junction, to generate transcripts containing three, two, and one guanosine at the 5' end, referred to as 3G, 2G, and 1G RNA, respectively. The 1G RNA is preferentially selected for packaging, indicating that these 99.9% identical RNAs exhibit functional differences and highlighting the importance of TSS selection. Here, we demonstrate that TSS selection is regulated by sequences between the CATA/TATA box and the beginning of R. Furthermore, we have generated two HIV-1 mutants with distinct 2-nucleotide modifications that predominantly express 3G RNA or 1G RNA. Both mutants can generate infectious viruses and undergo multiple rounds of replication in T cells. However, both mutants exhibit replication defects compared to the wild-type virus. The 3G-RNA-expressing mutant displays an RNA genome-packaging defect and delayed replication kinetics, whereas the 1G-RNA-expressing mutant exhibits reduced Gag expression and a replication fitness defect. Additionally, reversion of the latter mutant is frequently observed, consistent with sequence correction by plus-strand DNA transfer during reverse transcription. These findings demonstrate that HIV-1 maximizes its replication fitness by usurping the TSS heterogeneity of host RNA Pol II to generate unspliced RNAs with different specialized roles in viral replication. The three consecutive guanosines at the junction of U3 and R may also maintain HIV-1 genome integrity during reverse transcription. These studies reveal the intricate regulation of HIV-1 RNA and complex replication strategy.

The authors investigated whether there is an association between angiotensin-converting-enzyme inhibitor (ACEI)/angiotensin II receptor blocker (ARB) users with hemorrhoids. A total of 21,670 ACEI users and 21,670 ARB users during 2000 - 2012 from a national health insurance database were included. Hemorrhoid incidences were monitored to the end of 2013. To examine the risk of hemorrhoids, the Cox proportional hazard model and the Kaplan-Meier method were used. The incidence rate of hemorrhoids in ARB users was 6.64 per 1000 person-years, which was higher than that of the ACEI users (5.48 per 1000 person-years). The adjusted hazard ratio of hemorrhoids in patients who received ACEI relative to those who received ARB was 0.83 (95% confidence intervals [CI] = 0.75, 0.92). Compared to ARB users, patients who used ACEI more than 740 days per year (adjusted hazard ratios = 0.44; 95% CI = 0.36, 0.54) and more than 7800 mg (adjusted hazard ratios = 0.54; 95% CI = 0.46, 0.65) had a lower risk of hemorrhoids. ACEI users who took a relatively long period or high cumulative dosage were found to less prone to develop hemorrhoids., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc.)

Objective: This study focused on the predictive ability of the 3 scores for all-cause mortality in 6444 patients with atrial fibrillation (AF)., Methods: To assess the predictive accuracy of risk of death modelled by HATCH, HAVOC and CHA2DS2-VASc scores, the area under the curve of receiver operating characteristics (AUROC) was applied., Results: Over follow-up time, the cumulative incidence of death was clearly associated with the three scores (log-rank test, p<0.001). The AUROC for the HATCH (0.6618) was significantly higher than HAVOC Score (0.5733) and CHA2DS2-VAScs Score (0.6423)., Conclusions: HATCH score has better ability in predicting mortality in comparison to other two scores in patients with AF., (© The Author(s) 2022. Published by Oxford University Press on behalf of Postgraduate Medical Journal. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Purpose: The study compared the incidence of cancer between patients with diabetes with and without sodium-glucose cotransporter-2 (SGLT2) inhibitors use., Methods: This study identified a non-SGLT2 inhibitor cohort of 325,989 patients and a SGLT2 inhibitor cohort of 325,990 patients. The primary interest of this study was the occurrence of cancer. Hazard ratios (HRs) and 95 % confidence intervals (CIs) were estimated using Cox proportional hazard models., Results: Patients receiving SGLT2 inhibitors (adjusted HR = 0.79, 95 % CI = 0.76-0.83) had a significantly lower risk of developing cancer than patients without receiving SGLT2 inhibitors., Conclusion: The results demonstrated that patients with diabetes receiving SGLT2 inhibitors had a significantly lower risk of cancer., Competing Interests: Declaration of competing interest None., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Aim: This retrospective analysis aims to explore the risk of chronic kidney disease (CKD) among type 2 diabetes mellitus (DM) patients with different scores of adapted diabetes complications severity index (DCSI) who received sodium-glucose cotransporter-2 inhibitors (SGLT2Is)., Methods: This study includes 113,449 DM patients from the Taiwan National Health Insurance Research Database (NHIRD). We analyzed the data collected from 107,440 patients showing a DCSI score change of < 1 per year, 3720 patients with a score change of 1 to 2 per year and 2289 patients with a score change of > 2 per year. Cox proportional hazard models were used to evaluate the CKD risk throughout the overall follow-up period, and were adjusted for sex, age, comorbidities and medications of a-glucosidase inhibitors, angiotensin-converting enzyme inhibitors or angiotensin receptor blockers, biguanides, dipeptidyl peptidase-4 inhibitors, glucagon like peptide-1 receptor agonists, insulin, meglitinides, sulphonylurea and thiazolidinedione., Results: The incidence of CKD increased from 18.30 per 1000 person-years in patients with a score change of < 1 per year to 137.55 per 1000 person-years for those with a score change of > 2 per year. Patients with a higher score change (> 2 per year) and receiving SGLT2Is had a lower risk of developing CKD than patients who did not receive SGLT2Is., Conclusion: The use of SGLT2Is was significantly associated with the reduction in CKD incidence in diabetic patients with a higher DCSI., (© 2022. The Author(s) under exclusive licence to Italian Society of Nephrology.)

Objective: The current study detects the effect of sodium-glucose cotransporter-2 inhibitor (SGLT2I) versus beta-blocker (BB) in diabetes mellitus (DM) with chronic hepatitis B or C on hepatocellular carcinoma (HCC) outcomes. Methods: The multivariate logistic regression model, including all baseline characteristics and index year, was used to calculate the propensity scores, and we performed the greedy algorithm on propensity scores to create matched pairs of SGLT2I and BB users. Hazard ratios (HRs) and the corresponding 95% confidence intervals (CIs) of HCC were estimated by Cox proportional hazards regression models, and we adjusted for confounding factors by including the baseline characteristics in the regression models. Results: After matching in a ratio of 1:1, 7023 SGLT2I users and 7023 BB users were included in the following statistical analyses. The overall HRs showed a significantly lower risk of HCC in SGLT2I users in comparison to a reference group of BB users with an adjusted HR of 0.27 (0.21, 0.34). Conclusions: Compared to BB use, SGLT2I was associated with a significant risk reduction in HCC occurrence.

Background: A growing number of cytogenetic techniques have been used for prenatal diagnosis. This study aimed to demonstrate the usefulness of karyotyping, BACs-on-Beads (BoBs) assay and single nucleotide polymorphism (SNP) array in prenatal diagnosis during the second trimester based on our laboratory experience., Methods: A total of 10,580 pregnant women with a variety of indications for amniocentesis were enrolled in this retrospective study between January 2015 and December 2020, of whom amniotic fluid samples were analysed in 10,320 women. The main technical indicators of participants in the three different technologies were summarized, and cases of chromosome abnormalities were further evaluated., Results: The overall abnormality detection rate of karyotyping among all the amniotic fluid samples was 15.4%, and trisomy 21 was the most common abnormality (20.9%). The total abnormality detection rate of the BoBs assay was 5.6%, and the diagnosis rate of microdeletion/microduplication syndromes that were not identified by karyotyping was 0.2%. The detection results of the BoBs assay were 100.0% concordant with karyotyping analysis in common aneuploidies. Seventy (87.5%) cases of structural abnormalities were missed by BoBs assay. The total abnormality detection rate of the SNP array was 21.6%. The detection results of common aneuploidies were exactly the same between SNP array and karyotyping. Overall, 60.1% of structural abnormalities were missed by SNP array. The further detection rate of pathogenic significant copy number variations (CNVs) by SNP was 1.4%., Conclusions: Karyotyping analysis combined with BoBs assay or SNP array for prenatal diagnosis could provide quick and accurate results. Combined use of the technologies, especially with SNP array, improved the diagnostic yield and interpretation of the results, which contributes to genetic counselling. BoBs assay or SNP array could be a useful supplement to karyotyping., (© 2023. The Author(s).)

An important quality attribute of a recombinant adeno-associated virus (rAAV) as a therapeutic vector is its infectivity. Current assays to quantify infectious rAAV rely on coinfection with a helper virus such as adenovirus (Ad), which requires helper virus preparation and introduces additional variability. A simple method that has high sensitivity and removes the need for helper virus would improve assay consistency and facilitate high-throughput applications such as rAAV producer cell line development. In this study, we describe a stable assay cell line that was generated by integrating the coding sequences for AAV Rep68 and Ad E4orf6 and DNA binding protein under the control of inducible promoters. The Rep68 protein expression was further modulated by a ligand-responsive destabilization domain. In several benchmarks, the cell line gave comparable titers with those obtained using a classical Ad coinfection method. The cell line was also used to titer vectors of multiple rAAV serotypes. This cell line has the potential to serve as an effective and robust tool for quantifying infectious rAAV titers to advance gene therapy vector biomanufacturing.

Over the last two decades, model-based metabolic pathway optimization tools have been developed for the design of microorganisms to produce desired metabolites. However, few have considered more complex cellular systems such as mammalian cells, which requires the use of nonlinear kinetic models to capture the effects of concentration changes and cross-regulatory interactions. In this study, we develop a new two-stage pathway optimization framework based on kinetic models that incorporate detailed kinetics and regulation information. In Stage 1, a set of optimization problems are solved to identify and rank the enzymes that contribute the most to achieving the metabolic objective. Stage 2 then determines the optimal enzyme interventions for specified desired numbers of enzyme adjustments. It also incorporates multi-scenario optimization, which allows the simultaneous consideration of multiple physiological conditions. We apply the proposed framework to find enzyme adjustments that enable a reverse glucose flow in cultured mammalian cells, thereby eliminating the need for glucose feed in the late culture stage and enhancing process robustness. The computational results demonstrate the efficacy of the proposed approach; it not only captures the important regulations and key enzymes for reverse glycolysis but also identifies differences and commonalities in the metabolic requirements for different carbon sources., (© 2022 Wiley Periodicals LLC.)