Application of a single-cell-RNA-based biological-inspired graph neural network in diagnosis of primary liver tumors.

Single-cell technology depicts integrated tumor profiles including both tumor cells and tumor microenvironments, which theoretically enables more robust diagnosis than traditional diagnostic standards based on only pathology. However, the inherent challenges of single-cell RNA sequencing (scRNA-seq) data, such as high dimensionality, low signal-to-noise ratio (SNR), sparse and non-Euclidean nature, pose significant obstacles for traditional diagnostic approaches. The diagnostic value of single-cell technology has been largely unexplored despite the potential advantages. Here, we present a graph neural network-based framework tailored for molecular diagnosis of primary liver tumors using scRNA-seq data. Our approach capitalizes on the biological plausibility inherent in the intercellular communication networks within tumor samples. By integrating pathway activation features within cell clusters and modeling unidirectional inter-cellular communication, we achieve robust discrimination between malignant tumors (including hepatocellular carcinoma, HCC, and intrahepatic cholangiocarcinoma, iCCA) and benign tumors (focal nodular hyperplasia, FNH) by scRNA data of all tissue cells and immunocytes only. The efficacy to distinguish iCCA from HCC was further validated on public datasets. Through extending the application of high-throughput scRNA-seq data into diagnosis approaches focusing on integrated tumor microenvironment profiles rather than a few tumor markers, this framework also sheds light on minimal-invasive diagnostic methods based on migrating/circulating immunocytes.

Tumor budding in pre-neoadjuvant biopsy and post-neoadjuvant resection specimens is associated with poor prognosis in intrahepatic cholangiocarcinoma-a cohort study of 147 cases by modified ITBCC criteria.

Tumor budding (TB) has been associated with poor survival in a variety of cancers including intrahepatic cholangiocarcinoma (iCCA). As tumor histomorphological features are significantly altered after neoadjuvant therapy (NAT), our study aims to assess the prognostic significance of TB in iCCA patients before and after NAT, by the modified International Tumor Budding Consensus Conference (ITBCC) criteria. 147 NAT-treated iCCA cases were included in this study. In biopsy specimens obtained before NAT, the TB-positive subgroup had lower overall survival (OS) in univariate analysis (P = 0.010). In resection specimens obtained after NAT, the TB-positive subgroup had reduced OS (P = 0.002) and recurrence-free survival (RFS) (P = 0.013) in univariate analysis. In multivariate analysis including TNM stage, lymphovascular invasion and perineural invasion, TB-positive in post-NAT resection was also found as an independent prognostic factor for both OS and RFS (OS, HR, 3.005; 95% CI, 1.333-6.775, P = 0.008; RFS, HR, 1.748; 95% CI, 1.085-2.816, P = 0.022). In conclusion, assessing the presence of TB by modified ITBCC criteria provides robust prognostic information in the NAT setting of iCCA patients and can be considered to be included in routine pathological reporting.

Adsorption of cytochrome c on different self-assembled monolayers: The role of surface chemistry and charge density.

In this work, the adsorption behavior of cytochrome c (Cyt-c) on five different self-assembled monolayers (SAMs) (i.e., CH3-SAM, OH-SAM, NH2-SAM, COOH-SAM, and OSO3--SAM) was studied by combined parallel tempering Monte Carlo and molecular dynamics simulations. The results show that Cyt-c binds to the CH3-SAM through a hydrophobic patch (especially Ile81) and undergoes a slight reorientation, while the adsorption on the OH-SAM is relatively weak. Cyt-c cannot stably bind to the lower surface charge density (SCD, 7% protonation) NH2-SAM even under a relatively high ionic strength condition, while a higher SCD of 25% protonation promotes Cyt-c adsorption on the NH2-SAM. The preferred adsorption orientations of Cyt-c on the negatively-charged surfaces are very similar, regardless of the surface chemistry and the SCD. As the SCD increases, more counterions are attracted to the charged surfaces, forming distinct counterion layers. The secondary structure of Cyt-c is well kept when adsorbed on these SAMs except the OSO3--SAM surface. The deactivation of redox properties for Cyt-c adsorbed on the highly negatively-charged surface is due to the confinement of heme reorientation and the farther position of the central iron to the surfaces, as well as the relatively larger conformation change of Cyt-c adsorbed on the OSO3--SAM surface. This work may provide insightful guidance for the design of Cyt-c-based bioelectronic devices and controlled enzyme immobilization.

A bibliometric analysis of respiratory mechanics research related to acute respiratory distress syndrome from 1985 to 2023.

Background: Acute respiratory distress syndrome (ARDS) is a severe condition characterized by lung stiffness and compromised gas exchange, often requiring mechanical ventilation for treatment. In addition to its clinical significance, understanding the publication trends and research patterns in respiratory mechanics related to ARDS can provide insights into the evolution of this field from a bibliometric perspective, aiding in strategic planning and resource allocation for future research endeavors. Objective: This study aimed to explore the trends and identify the hotspots in respiratory mechanics research related to ARDS. Methods: All relevant studies on respiratory mechanics of ARDS published between 1985 and 2023 were retrieved from the Web of Science Core Collection (WoSCC), and the retrieval strategy was topic search "TS = respiratory mechanics OR lung mechanics AND TS = ARDS OR acute respiratory distress syndrome." Annual trends, citation patterns, and contributions from countries, institutions, authors, and journals were analyzed using Bibliometrix Biblioshiny. Networks and overlay of authors, institutions, countries, journals, co-citations, and keywords were analyzed and visualized using VOSviewer. Results: Our analysis included 1,248 articles published between 1985 and 2023, revealing fluctuations in publication output over time. The United States emerged as the leading contributor, with Critical Care Medicine being the most prominent journal. Key research themes included mechanical ventilation, acute lung injury, and protective ventilation strategies. International collaboration was evident, facilitating knowledge exchange and interdisciplinary cooperation. Conclusion: Our study sheds light on the evolving landscape of respiratory mechanics research in ARDS. International collaboration is pivotal in advancing the field, while researchers increasingly focus on personalized approaches to address the complexities of ARDS respiratory mechanics.

Long-term effects of vagus nerve stimulation on EEG aperiodic components in patients with drug-resistant epilepsy.

Background: Drug-resistant epilepsy (DRE) affects approximately one-third of epilepsy patients who do not achieve adequate seizure control with medication. Vagus nerve stimulation (VNS) is an adjunctive therapy for DRE, but its long-term effects on cortical excitability remain unclear. Objectives: This study aims to elucidate the long-term effects of VNS on electroencephalography (EEG) aperiodic components in patients with DRE. Our objective is to identify biomarkers that can serve as indicators of therapeutic efficacy and provide mechanistic insights into the underlying neural processes. Design: This longitudinal observational study focused on patients with DRE undergoing VNS therapy at Sanbo Brain Hospital. The reduction in seizure frequency rates was quantified over short-term (⩽1 year), medium-term (1-3 years), and long-term (⩾3 years) intervals to assess the therapeutic efficacy of VNS. Both the periodic and aperiodic components of EEG data were analyzed. Methods: Advanced signal processing techniques were utilized to parameterize the periodic and aperiodic components of EEG data, focusing particularly on "offset" and "exponent." These measures were compared before and after VNS therapy. Correlation analyses were conducted to explore the relationship between these EEG parameters and clinical outcomes. Results: In all, 18 patients with DRE participated in this study. During the long-term follow-up period, the responder rate was 55.56%. Significant decreases were observed in aperiodic offset (p = 0.022) and exponent (p = 0.039) among responders. The impact of age on these results was not significant. Correlation analyses revealed a negative association between therapeutic efficacy and a decrease in offset (R = -0.546, p = 0.019) and exponent (R = -0.636, p = 0.019). Conclusion: EEG aperiodic parameters, including offset and exponent, have the potential to serve as promising biomarkers for evaluating the efficacy of VNS. An understanding of the regulatory influence of VNS on cortical excitability through these aperiodic parameters could provide a basis for the development of more effective stimulation parameters and therapeutic strategies.

Monitoring Gene Sequences of Staphylococcus aureus Using a Love-Mode Surface Acoustic Wave Biosensor Coated with Cellulose Acetate/Polyethylenimine Nanofibers and Au Nanoparticles.

Love-mode surface acoustic wave (SAW) sensors show great promise for biodetection applications owing to their low cost, digital output, and wireless passive capability, but their performance is often restricted by the availability of suitable sensitive membrane layers. Herein, a composite layer of electrospun fibers made from cellulose acetate and polyethylenimine, coated with gold nanoparticles, is proposed as a porous and sensitive membrane coated onto a love-mode SAW biosensor for monitoring gene sequences of Staphylococcus aureus. The results showed that the developed sensor exhibited an impressive sensitivity of 122.56 Hz/(nmol/L) for detecting gene sequences of S. aureus, surpassing the sensitivity of conventional SAW sensors employing a bare Au film as the sensitive layer by 5-fold. The analysis revealed a remarkably linear detection (R2 of 0.97827) of S. aureus gene sequences within the range of 0 to 100 nmol/L. The limit of detection was impressively low at 0.9116 nmol/L. The good stability and specificity of the biosensor in liquid environments were demonstrated for clinical diagnostics.

Unraveling the orientation of an enzyme adsorbed onto a metal-organic framework.

Bio-conversion of lignocellulosic biomass to bioethanol fuel is a highly desirable yet challenging objective because of the low catalytic activity and high cost of ß-glucosidase (BGL). Recently, ZIF-8, an emerging organic porous material, has been proposed as a promising candidate for enzyme immobilization to improve associated activity and stability. However, the underlying interaction mechanism of binding BGL on the ZIF-8 surface is yet to be clarified. Here, the adsorption of BGL onto ZIF-8 is explored for the first time by molecular dynamics simulations. The results show that BGL adsorbs on the ZIF-8 surface with a "back-on" orientation. The adsorption free energy analysis shows that the adsorption process is enthalpy driven. In addition, the electrostatic interaction between negatively charged residues and Zn2+ on the surface of ZIF-8 is found to play a decisive role in surface binding, which accounts for 98% of the total interaction energy. The secondary structure of BGL is not affected despite the strong adsorption, suggesting the good biocompatibility of ZIF-8. This study not only provides a reliable theoretical insight into understanding the interaction mechanism between BGL and ZIF-8, but also helps the rational design of ZIF-8-based materials for bio-related applications.

Cocreation of social value: A tripartite game analysis with Chinese state-owned enterprises as an example.

In the context of the digital economy era, it is urgent for Chinese state-owned enterprises (SOEs) to engage in social value cocreation activities. The government and consumers' roles in SOEs' social value cocreation system cannot be ignored. Therefore, it is necessary to explore the tripartite social value cocreation model involving the government, SOEs, and consumers. In this respect, this study constructs a tripartite evolutionary game model of the government, SOEs, and consumers, and explores the influencing factors and evolutionary mechanism of the system overall. Matlab software is used to analyze the simulation data. The results reveal that the prerequisite for SOEs' successful social value cocreation is that consumers receive additional social value benefits greater than the level of improvement in social welfare. The allocation coefficient of consumers' additional social value benefits, the degree of the government's digital empowerment subsidy, and the level of the punishment for SOEs that violate the government's cocreation requirements will accelerate the achievement of equilibrium in the social value cocreation system, without affecting the final equilibrium result. By analyzing the strategic choices and interactive relationships among the government, SOEs, and consumers in social value cocreation in-depth, this study offers suggestions to promote the government, SOEs, and consumers' participation in social value cocreation. This research contributes to clarifying SOEs' social value cocreation model and has significant implications for promoting enterprises' high-quality development.

An Infrequent 1D Manganese-Tin Selenide with Bifunctional Catalytic Properties.

The optoelectronic, electrocatalytic, and photocatalytic properties of new tin selenides are of great significance for various energy applications. Herein, a facile solvothermal reaction of Sn, Se, and MnCl2 in 1,3-propodiamine (1,3-dap) solution at 150 °C for 7 days was used to achieve a new type of one-dimensional (1D) organic hybrid manganese-tin selenide [Mn2(1,3-dap)4(µ-1,3-dap)Sn2Se6]n (MnSnSe-1), whose 1D framework is built up from the linkage of rare unsaturated binuclear [Mn2(1,3-dap)4(µ-1,3-dap)]2+ cations and dimeric [Sn2Se6]4- anions. The combination of MnSnSe-1 and Ni nanoparticle is first applied for the preparation of a Ni/MnSnSe-1/NF electrode (NF = porous Ni foam) as the efficient electrocatalyst for the hydrogen evolution reaction (HER), indicating excellent HER electrocatalytic property with an overpotential of 117 mV at 10 mA·cm-2 in a neutral medium. Owing to its narrow absorption edge of 1.65 eV, implying prominent harvesting ability in the visible-light region, MnSnSe-1 shows a remarkable photocurrent response and excellent visible-light-driven photocatalytic property for the degradation of methylene blue in aqueous solution.

Clinical outcomes of three haploidentical transplantation protocols for hematologic malignancies based on data from the Chinese Bone Marrow Transplantation Registry Group.

This study aimed to demonstrate the clinical outcomes of granulocyte colony-stimulating factor (G-CSF)/antithymocyte globulin (ATG), posttransplantation cyclophosphamide (PTCy) and PTCy combined with lowdose ATG (PTCy with ATGlow)-based haploidentical transplantation protocols in patients with haematologic malignancies. The comparisons were conducted via propensity score matching (PSM) analysis to balance the basic characteristics among different groups and were based on the transplantation data reported to the Chinese Bone Marrow Transplantation Registry Group (CBMTRG) from January 2020 to December 2022. For each patient in the PTCy or PTCy with ATGlow group, patients (at a 1:2 ratio) from the GCSF/ ATG group were selected. In total, the PTCy group (n=122) was matched with G-CSF/ATG Group 1 (n=230), and the PTCy+ATGlow group (n=123) was matched with G-CSF/ATG Group 2 (n=226). Compared with those in the PTCy group, the incidences of 28-day neutrophil engraftment (P=0.005), 100- day platelet engraftment (P=0.002), median time to neutrophil engraftment (P.

Hypoglycemic drugs, circulating inflammatory proteins, and gallbladder diseases: A mediation mendelian randomization study.

BACKGROUND: The relationship of hypoglycemic drugs, inflammatory proteins and gallbladder diseases remain unknown. METHODS: Four hypoglycemic drugs were selected as exposure: glucagon-like peptide-1 receptor agonists (GLP-1RA), dipeptidyl peptidase-4 inhibitors (DPP-4i), sodium-glucose cotransporter 2 inhibitors (SGLT-2i), and metformin. The outcome were two gallbladder diseases: cholecystitis and cholelithiasis. Mendelian Randomization (MR) was employed to determine the association between hypoglycemic drugs and gallbladder diseases. RESULTS: DPP-4i and SGLT-2i had no effect on cholecystitis and cholelithiasis. However, a causal relationship was found between inhibition of ETFDH gene, a target of metformin expressed in cultured fibroblasts, and cholelithiasis (OR: 0.84, 95 %CI: (0.72,0.97), p = 0.021), as well as between GLP1R expression in the brain caudate basal ganglia and cholecystitis (OR: 1.29, 95 %CI: (1.11,1.49), p = 0.001). The effect of ETFDH inhibition on cholelithiasis through Interleukin-10 receptor subunit beta (IL-10RB) levels and Neurotrophin-3 (NT-3) levels, with a mediated proportion of 8 % and 8 %, respectively. CONCLUSION: Metformin plays a protective role in cholelithiasis, while GLP-1RA have a harmful effect on the risk of cholecystitis. Metformin may reduce the risk of cholelithiasis by modulating the levels of Neurotrophin-3 (NT-3) and Interleukin-10 receptor subunit beta (IL-10RB). Further clinical and mechanistic studies are required to confirm these findings.

Facilitating integrative and personalized oncology omics analysis with UCSCXenaShiny.

The continuous generation of multi-omics and phenotype data is propelling advancements in precision oncology. UCSCXenaShiny was developed as an interactive tool for exploring thousands of cancer datasets available on UCSC Xena. However, its capacity for comprehensive and personalized pan-cancer data analysis is being challenged by the growing demands. Here, we introduce UCSCXenaShiny v2, a milestone update through a variety of improvements. Firstly, by integrating multidimensional data and implementing adaptable sample settings, we create a suite of robust TPC (TCGA, PCAWG, CCLE) analysis pipelines. These pipelines empower users to conduct in-depth analyses of correlation, comparison, and survival in three modes: Individual, Pan-cancer and Batch screen. Additionally, the tool includes download interfaces that enable users to access diverse data and outcomes, several features also facilitate the joint analysis of drug sensitivity and multi-omics of cancer cell lines. UCSCXenaShiny v2 is an open-source R package and a web application, freely accessible at https://github.com/openbiox/UCSCXenaShiny .

Impact of Duration of Adjuvant Therapy on Patients with Initially Unresectable Hepatocellular Carcinoma After Conversion Surgery: A Propensity Score Matching Study.

Background: This study aimed to assess the effect of adjuvant therapy with different durations in patients with initially unresectable hepatocellular carcinoma (uHCC) after conversion surgery. Methods: This study included 85 patients with initially uHCC who received conversion surgery between May 2019 and November 2022. They were divided into the long duration group (n = 57) and short duration group (n = 28) based on postoperative medication duration. Recurrence-free survival (RFS) and overall survival (OS) were analyzed and compared between the cohorts. Results: No significant difference in RFS or OS was found between the two groups [RFS: hazard ratio (HR) = 0.486; 95% confidence interval (CI), 0.229-1.034, P = 0.061; OS: HR = 0.377; 95% CI, 0.119-1.196, P = 0.098]. Patients without major pathologic response (MPR) in the long duration group had better RFS and OS results compared to those in the short duration group (RFS: HR = 0.242; 95% CI, 0.092-0.634, P = 0.004; OS: HR = 0.264; 95% CI, 0.079-0.882, P = 0.031). No significant difference was detected in RFS or OS between the two groups in patients with MPR (RFS: HR = 1.250; 95% CI, 0.373-4.183, P = 0.718; OS: HR = 7.389; 95% CI, 0.147-372.4, P = 0.317). After propensity score matching, 25 pairs of patients were selected and the results remained consistent. Conclusion: At least 6 months of adjuvant therapy may be beneficial for patients without MPR after conversion surgery. However, in patients with MPR, the effect of adjuvant therapy remains unclear. Further studies are needed to confirm the optimal duration of adjuvant therapy.

Correlation between insulin-like growth factor and complexity of glucose time series index in patients with newly diagnosed acromegaly: a PILOT study.

BACKGROUND: Acromegaly has a high risk of abnormal glucose metabolism. The complexity of the glucose time series index (CGI) is calculated from refined composite multi-scale entropy analysis of the continuous glucose monitoring (CGM) data. CGI is a new indicator of glucose imbalance based on ambulatory glucose monitoring technology, which allows for earlier response to glucose metabolism imbalance and correlates with patient prognosis. OBJECTIVE: To compare the differences in glucose metabolic profile and CGI between acromegaly with normal glucose tolerance (NGT) and healthy subjects. METHODS: Eight newly diagnosed patients with acromegaly (GH group) and eight age- and gender-matched healthy subjects (Control group) were included in this study. All participants underwent oral glucose tolerance test (OGTT) and 72-h CGM. A refined composite multi-scale entropy analysis was performed on the CGM data to calculate the CGI and we compare the differences in glycemic profiles and CGI between the two groups. RESULTS: After OGTT, compared with the control group, patients in the GH group had higher 2 h blood glucose (BG) (mmol/L) [GH vs control, 6.7 (6.1, 7.0) vs 5.2 (3.8, 6.3), P = 0.012], 3 h BG [5.1 (3.8, 6.5) vs 4.0 (3.4, 4.2), P = 0.046], mean BG [6.3 (6.1, 6.5) vs 5.5 (5.1, 5.9), P = 0.002], 2 h insulin (mU/L) [112.9 (46.8, 175.5) vs 34.1 (17.1, 55.6), P = 0.009], and 3 h insulin [26.8 (17.1, 55.4) vs 10.4 (4.2, 17.8), P = 0.016]. CGI was lower in the GH group [2.77 (1.92, 3.15) vs 4.2 (3.3, 4.8), P = 0.008]. Spearman's correlation analysis showed insulin-like growth factor (IGF) (r = -0.897, P < 0.001) and mean glucose (r = -0.717, P = 0.003) were significantly negatively correlated with CGI. Multiple linear stepwise regression showed that IGF-1 (r = -0.652, P = 0.028) was independent factor associated with CGI in acromegaly. CONCLUSION: IGF-1 was significantly associated with CGI, and CGI may serve as a novel marker to evaluate glucose homeostasis in acromegaly with normal glucose tolerance.

The effect of tempering temperature on microstructure and corrosion resistance of M390 powder metallurgical martensitic stainless steel.

The corrosion resistance of M390 powder metallurgical martensitic stainless steel with different tempering temperatures was investigated by potentiodynamic polarization measurements, salt spray tests, and microstructural analyses utilizing scanning electron microscopy (SEM), X-ray diffraction (XRD), and transmission electron microscopy (TEM). The tempering temperature had no significant effect on the size and volume fraction of carbides. The corrosion resistance of M390 steel gradually deteriorated with increasing tempering temperature, and a loss passivation (LOP) effect was observed when tempered at 450 °C, 500 °C, and 550 °C. Transmission electron microscopy (TEM) analysis showed that the width of the Cr-depleted zones around the undissolved M7C3 carbides increased with increasing tempering temperature, while the Cr content in these zones decreased, which was the main reason for the deterioration of corrosion resistance. This study offers valuable insights into optimizing the tempering process to improve the corrosion resistance of M390 steel for practical applications.

Effect of family history of cancer on postoperative survival in patients with non-small cell lung cancer.

Background: Family history of cancer (FHC) has been reported to increase mortality of non-small cell lung cancer, mainly comprised of lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). However, the impact of FHC on long-term survival remains controversial. This study aims to identify the impact of FHC on postoperative survival in LUAD and LUSC. Methods: Patients underwent lung resection for LUAD or LUSC in West China Hospital from 2009 to 2021 were enrolled. The 5-year overall survival (OS), lung cancer-specific survival (LCSS) and progression-free survival (PFS) were compared between the patients with and without FHC. Multivariable Cox regression was also performed. Results: A total of 6,253 patients were enrolled, including 5,685 LUAD and 568 LUSC. Altogether 18.9% (1,077/5,685) patients had FHC in LUAD, and 12.7% (72/568) patients had FHC in LUSC. In LUAD, the patients with FHC showed comparable survival compared with the patients without FHC regarding 5-year OS (87.9% vs. 86.5%, P=0.49), 5-year PFS (84.8% vs. 80.9%, P=0.06), and 5-year LCSS (89.2% vs. 88.0%, P=0.96). In LUSC, the patients with FHC had poorer survival compared with the patients without FHC according to 5-year OS (40.9% vs. 68.2%, P=0.007), 5-year PFS (42.3% vs. 66.2%, P=0.003), and 5-year LCSS (45.8% vs. 72.7%, P=0.003). Multivariate analyses indicated that FHC was an independent prognostic factor of OS, PFS, and LCSS in the patients with LUSC. Conclusions: FHC was associated with a poor survival after lung resection in LUSC not LUAD patients. More attention should be paid in postoperative monitoring and treatment in LUSC patients with FHC.

A nomogram predicting the risk of extrathoracic metastasis at initial diagnosis of T≤3cmN0 lung cancer.

Background: The risk and risk factors of extrathoracic metastasis at initial diagnosis in T≤3cmN0 lung cancer patients are not fully understood. We aimed to develop a model to predict the risk of extrathoracic metastasis in those patients. Methods: Clinicopathological data of patients were collected from Surveillance, Epidemiology, and End Results (SEER) database. Univariable and multivariable analyses using logistic regression were conducted to identify risk factors. A predictive model and corresponding nomogram were developed based on the risk factors. The model was assessed using the area under the receiver operating characteristic curve (AUC), Hosmer-Lemeshow test, and decision curve. Results: A total of 20,057 T≤3cmN0 patients were enrolled, of whom 251 (1.25%) were diagnosed with extrathoracic metastasis at the initial diagnosis. Aged ≤50 [odds ratio (OR): 2.05, 95% confidence interval (CI): 1.19-3.53, P=0.01] and aged ≥81 [1.65 (1.05-2.58), P=0.03], Hispanic [1.81 (1.20-2.71), P=0.004], location of bronchus [3.18 (1.08-9.35), P=0.04], larger tumor size, pleural invasion, and a history of colorectal cancer [2.01 (1.01-4.00), P=0.046] were independent risk factors. In the training cohort and validation cohort, the AUCs of the developed model were 0.727, 0.728 respectively, and the results of Hosmer-Lemeshow test were P=0.47, P=0.61 respectively. The decision curve showed good clinical meaning of the model. Conclusions: Extrathoracic metastasis at initial diagnosis in T≤3cmN0 lung cancer patients was not rare. The model based on the risk factors showed good performance in predicting the risk of extrathoracic metastasis.

Evolved cytidine and adenine base editors with high precision and minimized off-target activity by a continuous directed evolution system in mammalian cells.

Continuous directed evolution of base editors (BEs) has been successful in bacteria cells, but not yet in mammalian cells. Here, we report the development of a Continuous Directed Evolution system in Mammalian cells (CDEM). CDEM enables the BE evolution in a full-length manner with Cas9 nickase. We harness CDEM to evolve the deaminases of cytosine base editor BE3 and adenine base editors, ABEmax and ABE8e. The evolved cytidine deaminase variants on BE4 architecture show not only narrowed editing windows, but also higher editing purity and low off-target activity without a trade-off in on-targeting activity. The evolved ABEmax and ABE8e variants exhibit narrowed or shifted editing windows to different extents, and lower off-target effects. The results illustrate that CDEM is a simple but powerful approach to continuously evolve BEs without size restriction in the mammalian environment, which is advantageous over continuous directed evolution system in bacteria cells.

Development of purity certified reference materials to establish metrological traceability for the measurement of nitroimidazoles in agricultural products.

It is of significant importance to public health that reliable monitoring of nitroimidazoles be conducted, while certified reference materials (CRMs) are essential for accurate and reliable detection. A project has been initiated with the objective of developing nitroimidazole purity CRMs to ensure that results from nationwide monitoring laboratories for nitroimidazoles in antibiotic residues can be compared and traced. The candidates were successively characterized in terms of their structure by means of infrared (IR) spectroscopy and mass spectrometry (MS). The mass balance (MB) method and the quantitative nuclear magnetic resonance (qNMR) method were utilized to determine the purity of nitroimidazoles with remarkable accuracy. Furthermore, a methodical investigation was conducted on homogeneity, stability, and uncertainty. Six nitroimidazole purity CRMs, including tinidazole (GBW09252), secnidazole (GBW09286), ronidazole (GBW09288), metronidazole (GBW(E)090755), dimetridazole (GBW(E)090819), and ornidazole (GBW(E)090820), were finally manufactured following authorization from China's State Administration for Market Regulation (SAMR). By using these CRMs, it is possible to improve the traceability, accuracy, and comparability of nitroimidazole measurements in a range of agricultural products, protecting public health.

Lung-Targeted Lipid Nanoparticle-Delivered siUSP33 Attenuates SARS-CoV-2 Replication and Virulence by Promoting Envelope Degradation.

As a structural protein of SARS-CoV-2, the envelope (E) protein not only plays a key role in the formation of viral particles, but also forms ion channels and has pathogenic functions, including triggering cell death and inflammatory responses. The stability of E proteins is controlled by the host ubiquitin-proteasome system. By screening human deubiquitinases, it is found that ubiquitin-specific protease 33 (USP33) can enhance the stability of E proteins depending on its deubiquitinase activity, thereby promoting viral replication. In the absence of USP33, E proteins are rapidly degraded, leading to a reduced viral load and inflammation. Using lipid nanoparticle (LNP) encapsulation of siUSP33 by adjusting the lipid components (ionizable cationic lipids), siUSP33 is successfully delivered to mouse lung tissues, rapidly reducing USP33 expression in the lungs and maintaining knockdown for at least 14 days, effectively suppressing viral replication and virulence. This method of delivery allows efficient targeting of the lungs and a response to acute infections without long-term USP33 deficiency. This research, based on the deubiquitination mechanism of USP33 on the E protein, demonstrates that LNP-mediated siRNA delivery targeting USP33 plays a role in antiviral and anti-inflammatory responses, offering a novel strategy for the prevention and treatment of SARS-CoV-2.