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Huang Lei,* Xingyou Dong,* Longkun Li,* Feng Huan, Xiao Zhong, Qingjian Wu, He Fang, Teng Zhang, Xinliang Yang, Jingzhen Zhu, Jia Li, Zhao Jiang Department of Urology, Second Affiliated Hospital, Army Military Medical University, Chongqing 400037, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhao JiangDepartment of Urology, Second Affiliated Hospital, Army Military Medical University, Chongqing 400037, People’s Republic of ChinaTel +86 23 68774624Email urologyzhaoj@sohu.comObjective: To explore the risk factors, pathogenic bacteria distribution and drug resistance of systematic transrectal ultrasound-guided prostate biopsy (TRUS-Bx), 329 cases of TRUS-Bx were collected, retrospectively, in the Second Affiliated Hospital, Army Military Medical University, from April 2017 to October 2019.Methods: A total of 329 cases were all qualified and grouped into the SIRS group (25 cases) and the non-SIRS group (304 cases). Of all the cases, incidence and risk factors of systemic inflammatory response syndrome (SIRS) were analyzed. Urine and blood samples of patients with SIRS after TRUS-Bx were also collected for bacterial culture and drug sensitivity test.Results: Multivariate logistic regression analysis showed that BMI ≥ 25 kg/m2 (OR = 1.66, 95% CI = 1.34– 2.12, P < 0.001), history of diabetes (OR = 5.48, 95% CI = 1.53– 19.68, P = 0.008), urinary infection before operation (OR = 9.19, 95% CI = 2.92– 20.93, P < 0.001) and erythrocyte sedimentation (ESR) ≥ 20 mm/h (OR = 1.04, 95% CI = 1.01– 1.08, P = 0.039) were independent risk factors of SIRS after TURS-PB.Conclusion: The incidence of SIRS and urinary sepsis was 7.59% and 2.13%, respectively, and major pathogens of SIRS after TRUS-Bx were Escherichia coli (58.33%), Klebsiella pneumoniae (12.5%) and Pseudomonas aeruginosa (12.5%). Imipenem, meropenem, tigecycline, piperacillin/tazobactam, teicoplanin, vancomycin, amikacin and cefoperazone/sulbactam had a very strong inhibitory effect to those pathogenic bacteria (sensitivity 85.72%∼ 100%). Levofloxacin, ciprofloxacin, gentamicin, penicillin G, compound neonomine and second-generation cephalosporins showed less but also worked as a good inhibitor to pathogenic bacteria (42.86%∼ 80.95%).Keywords: systematic transrectal ultrasound-guided prostate biopsy, systemic inflammatory response syndrome, prostate cancer, risk factors infection, pathogens

Abstract In this study, a structural adhesive was used to bond unidirectional prepreg and fiber fabric in a single lap joint. The mechanical properties of the structural adhesive were investigated under room temperature dry state (RTD) and elevated temperature wet state (ETW, 71 ℃/85% RH), and different adhesive layer thicknesses (0.5 mm, 1.0 mm, 1.5 mm, and 2.0 mm). The fracture surfaces of the bonded joints were examined using scanning electron microscopy (SEM), and finite element simulations were conducted to observe the failure modes and failure paths. Additionally, the specimens were immersed in water and hydraulic oil, and their tensile shear strength was tested to evaluate their liquid sensitivity. The experimental results indicated that with increasing adhesive layer thickness, the strength of the specimens decreased by 21% in the RTD and by 52% in the ETW. The strength differences between different environments were minimal for adhesive layer thicknesses of 1 mm and 1.5 mm. The shear strength of the specimens decreased after immersion in water and hydraulic oil, with reductions of 43.78% and 39.21%, compared to the room temperature dry respectively. SEM observations of the bonded joint sections revealed that the primary failure modes were adherend failure and adhesive layer failure. Finite element simulations indicated that fiber tearing and crack initiation occurred in stress concentration areas during loading, leading to structural failure.

Abstract Background Respiratory infectious diseases have the highest incidence among infectious diseases worldwide. Currently, global monitoring of respiratory pathogens primarily focuses on influenza and coronaviruses. This study included influenza and other common respiratory pathogens to establish a local respiratory pathogen spectrum. We investigated and analyzed the co-infection patterns of these pathogens and explored the impact of lifting non-pharmaceutical interventions (NPIs) on the transmission of influenza and other respiratory pathogens. Additionally, we used a predictive model for infectious diseases, utilizing the commonly used An autoregressive comprehensive moving average model (ARIMA), which can effectively forecast disease incidence. Methods From June 2023 to February 2024, we collected influenza-like illness (ILI) cases weekly from the community in Xuanwu District, Nanjing, and obtained 2046 samples. We established a spectrum of respiratory pathogens in Nanjing and analysed the age distribution and clinical symptom distribution of various pathogens. We compared age, gender, symptom counts, and viral loads between individuals with co-infections and those with single infections. An autoregressive comprehensive moving average model (ARIMA) was constructed to predict the incidence of respiratory infectious diseases. Results Among 2046 samples, the total detection rate of respiratory pathogen nucleic acids was 53.37% (1092/2046), with influenza A virus 479 cases (23.41%), influenza B virus 224 cases (10.95%), and HCoV 95 cases (4.64%) being predominant. Some pathogens were statistically significant in age and number of symptoms. The positive rate of mixed infections was 6.11% (125/2046). There was no significant difference in age or number of symptoms between co-infection and simple infection. After multiple iterative analyses, an ARIMA model (0,1,4), (0,0,0) was established as the optimal model, with an R2 value of 0.930, indicating good predictive performance. Conclusions The spectrum of respiratory pathogens in Nanjing, Jiangsu Province, was complex in the past. The primary age groups of different viruses were different, causing various symptoms, and the co-infection of viruses did not correlate with the age and gender of patients. The ARIMA model estimated future incidence, which plateaued in subsequent months.

Genetically encoded circuits have been successfully utilized to assess and characterize target variants with desirable traits from large mutant libraries. Adenosylcobalamin is an essential coenzyme that is required in many intracellular physiological reactions and is widely used in the pharmaceutical and food industries. High-throughput screening techniques capable of detecting adenosylcobalamin productivity and selecting superior adenosylcobalamin biosynthesis strains are critical for the creation of an effective microbial cell factory for the production of adenosylcobalamin at an industrial level. In this study, we developed an RNA-protein hybrid biosensor whose input part was an endogenous RNA riboswitch to specifically respond to adenosylcobalamin, the inverter part was an orthogonal transcriptional repressor to obtain signal inversion, and the output part was a fluorescent protein to be easily detected. The hybrid biosensor could specifically and positively correlate adenosylcobalamin concentrations to green fluorescent protein expression levels in vivo. This study also improved the operating concentration and dynamic range of the hybrid biosensor by systematic optimization. An individual cell harboring the hybrid biosensor presented over 20-fold higher fluorescence intensity than the negative control. Then, using such a biosensor combined with fluorescence-activated cell sorting, we established a high-throughput screening platform for screening adenosylcobalamin overproducers. This study demonstrates that this platform has significant potential to quickly isolate high-productive strains to meet industrial demand and that the framework is acceptable for various metabolites.

Vitamin B12 is a complex compound synthesized by microorganisms. The industrial production of vitamin B12 relies on specific microbial fermentation processes. E. coli has been utilized as a host for the de novo biosynthesis of vitamin B12, incorporating approximately 30 heterologous genes. However, a metabolic imbalance in the intricate pathway significantly limits vitamin B12 production. In this study, we employed multivariate modular metabolic engineering to enhance vitamin B12 production in E. coli by manipulating two modules comprising a total of 10 genes within the vitamin B12 biosynthetic pathway. These two modules were integrated into the chromosome of a chassis cell, regulated by T7, J23119, and J23106 promoters to achieve combinatorial pathway optimization. The highest vitamin B12 titer was attained by engineering the two modules controlled by J23119 and T7 promoters. The inclusion of yeast powder to the fermentation medium increased the vitamin B12 titer to 1.52 mg/L. This enhancement was attributed to the effect of yeast powder on elevating the oxygen transfer rate and augmenting the strain's isopropyl-β-d-1-thiogalactopyranoside (IPTG) tolerance. Ultimately, vitamin B12 titer of 2.89 mg/L was achieved through scaled-up fermentation in a 5-liter fermenter. The strategies reported herein will expedite the development of industry-scale vitamin B12 production utilizing E. coli.

Abstract Background Bacillus subtilis is widely used in industrial-scale riboflavin production. Previous studies have shown that targeted mutagenesis of the ribulose 5-phosphate 3-epimerase in B. subtilis can significantly enhance riboflavin production. This modification also leads to an increase in purine intermediate concentrations in the medium. Interestingly, B. subtilis exhibits remarkable efficiency in purine nucleoside synthesis, often exceeding riboflavin yields. These observations highlight the importance of the conversion steps from inosine-5’-monophosphate (IMP) to 2,5-diamino-6-ribosylamino-4(3 H)-pyrimidinone-5’-phosphate (DARPP) in riboflavin production by B. subtilis. However, research elucidating the specific impact of these reactions on riboflavin production remains limited. Result We expressed the genes encoding enzymes involved in these reactions (guaB, guaA, gmk, ndk, ribA) using a synthetic operon. Introduction of the plasmid carrying this synthetic operon led to a 3.09-fold increase in riboflavin production compared to the control strain. Exclusion of gmk from the synthetic operon resulted in a 36% decrease in riboflavin production, which was further reduced when guaB and guaA were not co-expressed. By integrating the synthetic operon into the genome and employing additional engineering strategies, we achieved riboflavin production levels of 2702 mg/L. Medium optimization further increased production to 3477 mg/L, with a yield of 0.0869 g riboflavin per g of sucrose. Conclusion The conversion steps from IMP to DARPP play a critical role in riboflavin production by B. subtilis. Our overexpression strategies have demonstrated their effectiveness in overcoming these limiting factors and enhancing riboflavin production.

Abstract Biotin, serving as a coenzyme in carboxylation reactions, is a vital nutrient crucial for the natural growth, development, and overall well-being of both humans and animals. Consequently, biotin is widely utilized in various industries, including feed, food, and pharmaceuticals. Despite its potential advantages, the chemical synthesis of biotin for commercial production encounters environmental and safety challenges. The burgeoning field of synthetic biology now allows for the creation of microbial cell factories producing bio-based products, offering a cost-effective alternative to chemical synthesis for biotin production. This review outlines the pathway and regulatory mechanism involved in biotin biosynthesis. Then, the strategies to enhance biotin production through both traditional chemical mutagenesis and advanced metabolic engineering are discussed. Finally, the article explores the limitations and future prospects of microbial biotin production. This comprehensive review not only discusses strategies for biotin enhancement but also provides in-depth insights into systematic metabolic engineering approaches aimed at boosting biotin production.

ObjectiveTo analyze epidemiological characteristics and determinants of hand, foot, and mouth disease (HFMD) outbreaks in Jiangsu province over the period of 2010 to 2022 for developing local HFMD prevention and control measures. MethodsThe data on 238 HFMD outbreaks reported in Jiangsu province from 2010 through 2022 were collected from the Public Health Emergency Information Management System of the Chinese Center for Disease Control and Prevention. The epidemiological characteristics of the HFMD outbreaks were described using statistical analysis. The spatial distribution characteristics of the outbreaks were analyzed using ArcGIS10.7 and GeoDa1.18 software. The main influencing factors of the incidence and duration of the HFMD outbreaks were analyzed using a unconditional multivariate logistic regression model. ResultsA total of 238 outbreaks of HFMD were reported in the province during the study period, involving a total of 167 934 individuals exposed to the epidemic and resulting in 5 296 confirmed cases. The median attack rate for these outbreaks was 3.43%. The HFMD outbreaks were more frequently observed during the spring season (March to May) and autumn season (September to November), accounting for 36.97% and 29.83% of all outbreaks respectively. Furthermore, 68.07% of the outbreaks occurred in the southern region of the province. Almost all (99.16%) of these outbreaks took place in childcare institutions and schools. The time interval between incidence and identification ranged from 0 to 44 days (median = 8 days), while the outbreak duration varied from 3 to 78 days, with a median duration of 15.5 days. The predominant strains responsible for the HFMD outbreaks differed by year: Coxsackievirus A16 (Cox A16) was prevalent in 2010, 2012, 2014, 2015, 2018, 2019, and 2021; enterovirus 71 (EV71) dominated in 2011, 2013, and 2017; Coxsackievirus (Cox A6) was prominent in 2020; and both Cox A16 and EV71 were significant contributors in 2016; finally, Cox A16 and Cox A6 were identified as major strains in 2022. Geographically speaking, the HFMD outbreaks exhibited an aggregated distribution pattern with high-high clustering areas primarily located within three districts in Wuxi city, a district and a prefecture in Changzhou city, and three districts in Lianyungang city. Results obtained from multivariate logistic regression analysis indicated that certain characteristics were associated with higher attack rates among these outbreaks. These included being caused by Cox A16 or multiple viruses, having an epidemic duration lasting 21 days or longer, and being reported by schools, health centers, and education department. On the other hand, outbreaks occurring outside childcare institutions tended to have lower attack rates. Additionally, outbreaks classified as general epidemics, taking place in towns, and having a lag identification time of 15 days or more were more likely to have a longer epidemic duration. ConclusionThe HFMD outbreaks in Jiangsu province from 2010 to 2022 predominantly occurred during the spring and autumn seasons, with Cox A16 as the predominant strain. The southern region of Jiangsu province was the clustering area for the HFMD outbreaks. The main factors influencing the incidence and duration of these outbreaks include the outbreak setting, type of predominant virus, epidemic duration, reporting institution and epidemic level, location of the site, and lag time of epidemic identification.

ABSTRACTThe effectiveness of business process analysis heavily relies on the quality of event logs. However, the presence of outliers and missing values often compromises the integrity of event logs, consequently exerting adverse effects on process analysis and associated decision-making. Existing log repair research mainly focuses on the reconstruction of missing activity, whereas few efforts are carried out from the perspective of predicting missing activity. This paper introduces a log repair approach based on a masked Transformer, which innovatively combines the self-attention mechanism of Transformers with the task of event log repair. Firstly, by employing various masking strategies, we simulate diverse low-quality event log scenarios that may occur in practical situations. Subsequently, a Masked Language Model is trained on preprocessed datasets to predict masked activities by leveraging contextual information within traces, thereby capturing behavioral information of activities in the event log. Upon completion of model training, we apply it to real event log data for repair tasks. The proposed approach, originating from the perspective of event logs, does not rely on any a priori knowledge related to business process models for generating event logs. Experimental results demonstrate that the masked Transformer-based approach outperforms baseline methods in most event log repair tasks.

Immune cell infiltration and tumor-related immune molecules play key roles in tumorigenesis and tumor progression. The influence of immune interactions on the molecular characteristics and prognosis of clear cell renal cell carcinoma (ccRCC) remains unclear. A machine learning algorithm was applied to the transcriptome data from The Cancer Genome Atlas database to determine the immunophenotypic and immunological characteristics of ccRCC patients. These algorithms included single-sample gene set enrichment analyses and cell type identification. Using bioinformatics techniques, we examined the prognostic potential and regulatory networks of immune-related genes (IRGs) involved in ccRCC immune interactions. Fifteen IRGs (CCL7, CHGA, CMA1, CRABP2, IFNE, ISG15, NPR3, PDIA2, PGLYRP2, PLA2G2A, SAA1, TEK, TGFA, TNFSF14, and UCN2) were identified as prognostic IRGs associated with overall survival and were used to construct a prognostic model. The area under the receiver operating characteristic curve at 1 year was 0.927; 3 years, 0.822; and 5 years, 0.717, indicating good predictive accuracy. Molecular regulatory networks were found to govern immune interactions in ccRCC. Additionally, we developed a nomogram containing the model and clinical characteristics with high prognostic potential. By systematically examining the sophisticated regulatory mechanisms, molecular characteristics, and prognostic potential of ccRCC immune interactions, we provided an important framework for understanding the molecular mechanisms of ccRCC and identifying new prognostic markers and therapeutic targets for future research.

Predicting nutrient loads is essential to understanding and managing one of the environmental issues faced by the northern Gulf of Mexico hypoxic zone, which poses a severe threat to the Gulf’s healthy ecosystem and economy. The development of hypoxia in the Gulf of Mexico is strongly associated with the eutrophication process initiated by excessive nutrient loads. Due to the complexities in the excessive nutrient loads to the Gulf of Mexico, it is challenging to understand and predict the underlying temporal variation of nutrient loads. The study was aimed at identifying an optimal predictive machine learning model to capture and predict nonlinear behavior of the nutrient loads delivered from the Mississippi/Atchafalaya River Basin (MARB) to the Gulf of Mexico. For this purpose, monthly nutrient loads (N and P) in tons were collected from US Geological Survey (USGS) monitoring station 07373420 from 1980 to 2020. Machine learning models—including autoregressive integrated moving average (ARIMA), gaussian process regression (GPR), single-layer multilayer perceptron (MLP), and a long short-term memory (LSTM) with the single hidden layer—were developed to predict the monthly nutrient loads, and model performances were evaluated by standard assessment metrics—Root Mean Square Error (RMSE) and Correlation Coefficient (R). The residuals of predictive models were examined by the Durbin–Watson statistic. The results showed that MLP and LSTM persistently achieved better accuracy in predicting monthly TN and TP loads compared to GPR and ARIMA. In addition, GPR models achieved slightly better test RMSE score than ARIMA models while their correlation coefficients are much lower than ARIMA models. Moreover, MLP performed slightly better than LSTM in predicting monthly TP loads while LSTM slightly outperformed for TN loads. Furthermore, it was found that the optimizer and number of inputs didn’t show effects on the LSTM performance while they exhibited impacts on MLP outcomes. This study explores the capability of machine learning models to accurately predict nonlinearly fluctuating nutrient loads delivered to the Gulf of Mexico. Further efforts focus on improving the accuracy of forecasting using hybrid models which combine several machine learning models with superior predictive performance for nutrient fluxes throughout the MARB.

Abstract Intracellular proteome aggregation is a ubiquitous disease hallmark with its composition associated with pathogenicity. Herein, this work reports on a cell‐permeable photosensitizer (P8, Rose Bengal derivative) for selective photo induced proximity labeling and crosslinking of cellular aggregated proteome. Rose Bengal is identified out of common photosensitizer scaffolds for its unique intrinsic binding affinity to various protein aggregates driven by the hydrophobic effect. Further acetylation permeabilizes Rose Bengal to selectively image, label, and crosslink aggregated proteome in live stressed cells. A combination of photo‐chemical, tandem mass spectrometry, and protein biochemistry characterizations reveals the complexity in photosensitizing pathways (both Type I & II), modification sites and labeling mechanisms. The diverse labeling sites and reaction types result in highly effective enrichment and identification of aggregated proteome. Finally, aggregated proteomics and interaction analyses thereby reveal extensive entangling of proteostasis network components mediated by HSP70 chaperone (HSPA1B) and active participation of autophagy pathway in combating proteasome inhibition. Overall, this work exemplifies the first photo induced proximity labeling and crosslinking method (namely AggID) to profile intracellular aggregated proteome and analyze its interactions.

Omega-3 fatty acids are in high demand due to their efficacy in treating hypertriglyceridemia and preventing cardiovascular diseases. However, the growth of the industry is hampered by low purity and insufficient productivity. This study aims to develop an efficient RP-MPLC purification method for omega-3 fatty acid ethyl esters with high purity and capacity. The results indicate that the AQ-C18 featuring polar end-capped silanol groups outperformed C18 and others in retention time and impurity separation. By injecting pure fish oil esters with a volume equivalent to a 1.25% bed volume on an AQ-C18 MPLC column using a binary isocratic methanol–water (90:10, v:v) mobile phase at 30 mL/min, optimal omega-3 fatty acid ethyl esters were obtained, with the notable purity of 90.34% and a recovery rate of 74.30%. The total content of EPA and DHA produced increased from 67.91% to 85.27%, meeting the acceptance criteria of no less than 84% set by the 2020 edition of the Pharmacopoeia of the People’s Republic of China. In contrast, RP-MPLC significantly enhanced the production efficiency per unit output compared to RP-HPLC. This study demonstrates a pioneering approach to producing omega-3 fatty acid ethyl esters with high purity and of greater quantity using AQ-C18 RP-MPLC, showing this method’s significant potential for use in industrial-scale manufacturing.

Abstract Adenosylcobalamin (AdoCbl), a biologically active form of vitamin B12 (coenzyme B12), is one of the most complex metal-containing natural compounds and an essential vitamin for animals. However, AdoCbl can only be de novo synthesized by prokaryotes, and its industrial manufacturing to date was limited to bacterial fermentation. Here, we report a method for the synthesis of AdoCbl based on a cell-free reaction system performing a cascade of catalytic reactions from 5-aminolevulinic acid (5-ALA), an inexpensive compound. More than 30 biocatalytic reactions are integrated and optimized to achieve the complete cell-free synthesis of AdoCbl, after overcoming feedback inhibition, the complicated detection, instability of intermediate products, as well as imbalance and competition of cofactors. In the end, this cell-free system produces 417.41 μg/L and 5.78 mg/L of AdoCbl using 5-ALA and the purified intermediate product hydrogenobyrate as substrates, respectively. The strategies of coordinating synthetic modules of complex cell-free system describe here will be generally useful for developing cell-free platforms to produce complex natural compounds with long and complicated biosynthetic pathways.

Abstract Molecular rotor‐based fluorophores (RBFs) activate fluorescence upon increase of micro‐viscosity, thus bearing a broad application promise in many fields. However, it remains a challenge to control how fluorescence of RBFs responds to viscosity changes. Herein, we demonstrate that the formation and regulation of intramolecular hydrogen bonds in the excited state of RBFs could modulate their rotational barrier, leading to a rational control of how their fluorescence can be activated by micro‐viscosity. Based on this strategy, a series of RBFs were developed based on 4‐hydroxybenzylidene‐imidazolinone (HBI) that span a wide range of viscosity sensitivity. Combined with the AggTag method that we previously reported, the varying viscosity sensitivity and emission spectra of these probes enabled a dual‐color imaging strategy that detects both protein oligomers and aggregates during the multistep aggregation process of proteins in live cells. In summary, our work indicates that installing intracellular excited state hydrogen bonds to RBFs allows for a rational control of rotational barrier, thus allow for a fine tune of their viscosity sensitivity. Beyond RBFs, we envision similar strategies can be applied to control the fluorogenic behavior of a large group of fluorophores whose emission is dependent on excited state rotational motion, including aggregation‐induced emission fluorophores.

The rapid development of chip manufacturing technology has increased the demand for precise characterization techniques. The characterization technique of the physical field on the surface of a chip is crucial for analyzing chip failures and diagnosing faults. In this review, the latest advancements based on different measurement mechanisms are analyzed and summarized for the electromagnetic field characterization of the chip surfaces. In addition, their advantages and limitations are discussed. Finally, prospects for future development are presented.

Abstract Background Circular RNAs (circRNAs) may regulate the onset and progression of human malignancies by competitively binding to microRNA (miRNA) sponges, thus regulating the downstream genes. However, aberrant circRNA expression patterns and their biological functions in prostate cancer (PCa) warrant further studies. Our research sought to shed further light on the possible role and molecular mechanism of circEPHA3 action in controlling the growth and metastasis of PCa cells. Materials and methods circEPHA3 (has_circ_0066596) was initially screened from a previous circRNA microarray and identified following Actinomycin D and RNase R assays. Fluorescence in situ hybridization, biotin-coupled probe RNA pulldown, and dual-luciferase reporter gene assays were performed to examine the relationship between circEPHA3 and miR-513a-3p. The biological role of circEPHA3 in PCa was assessed by CCK8, wound healing, Transwell assays, and animal experiments. Results We identified a novel circular RNA, circEPHA3 (has_circ_0066596), which was down-regulated in high-grade PCa tissues and cell lines. The outcomes of CCK8, wound healing, Transwell assays, and animal experiments revealed that circEPHA3 prohibited the progression and metastasis of PCa in vivo and in vitro. Mechanistically, circEPHA3 was directly bound to miR-513a-3p and regulated the downstream gene, BMP2, thereby serving as a tumor suppressor in PCa. Conclusions As a tumor suppressor, circEPHA3 inhibited the proliferation and metastasis of PCa cells through the miR-513a-3p/BMP2 axis, suggesting that circEPHA3 might be a potential therapeutic target for PCa.

Introduction: Mantle cell lymphoma (MCL) can be stratified into blastoid and classical subtypes based on morphological features, with the former subtype having a poorer prognosis. Despite recent advances in targeted approaches, including multiple bruton tyrosine kinase inhibitors which yield impressive clinical responses and improve prognoses, MCL remains an incurable disease with frequent relapses. Additional therapeutic interventions are therefore unmet medical needs for the management of patients with MCL. Methods: Cell viability and apoptosis assays were employed to analyze the therapeutic interaction of venetoclax combined with selinexor in MCL cells. Western blot was used to investigate the potential mechanism of action for the synergy of venetoclax in combination with selinexor in MCL cells. Results: In this study, we revealed that both blastoid and classical MCL cells were vulnerable to the cytotoxic effects of selinexor, a well-established XPO1 inhibitor, manifested by loss of cell viability and induction of cell apoptosis. Moreover, our data indicated that the addition of venetoclax to selinexor showed synergistically decreased cell viabilities and increased cell deaths in blastoid and classical MCL cells compared to each single drug treatment. Either selinexor or venetoclax treatment alone decreased MCL1 expressions and increased BAX levels in MCL cells, and these effects were further enhanced by their combined regimen. Mechanistically, our findings demonstrated that induction of DNA damage and inactivation of DNA damage response were involved in the synergistic interaction of the drug combination regimen. Conclusion: Collectively, this study might provide a potential attractive therapy option for the treatment of MCL. However, the conclusion needs additional experimental validation in in vivo models and clinical evaluations are mandatory.

Anti-tumor drug resistance is a challenge for human triple-negative breast cancer (TNBC) treatment. Our previous work demonstrated that TNFAIP2 activates RAC1 to promote TNBC cell proliferation and migration. However, the mechanism by which TNFAIP2 activates RAC1 is unknown. In this study, we found that TNFAIP2 interacts with IQGAP1 and Integrin β4. Integrin β4 activates RAC1 through TNFAIP2 and IQGAP1 and confers DNA damage-related drug resistance in TNBC. These results indicate that the Integrin β4/TNFAIP2/IQGAP1/RAC1 axis provides potential therapeutic targets to overcome DNA damage-related drug resistance in TNBC.

In monomictic lakes, the potential intensification of hypolimnetic deoxygenation due to human activities and climate change threatens lake ecosystems and water quality. Understanding the trends and mechanisms of hypolimnetic deoxygenation would allow for the adaptive management of monomictic lakes. This study investigated the trends and drivers of hypolimnetic deoxygenation in Lake Fuxian (a monomictic lake in south-western China) over the past 150 years using paleolimnological evidence and historical records. Analysis of enrichment factors for molybdenum (Mo), uranium (U), and vanadium (V), and ratios of U/thorium (Th), pristane (Pr)/phytane (Ph), and total organic carbon (TOC)/total phosphorus (TP) in a dated sediment core indicated that the extent of hypolimnetic deoxygenation was relatively low before 1994 and increased substantially thereafter. Nutrient monitoring records and sediment n-alkane proxies indicated notable eutrophication of the lake since 1986 and a greater degree of human disturbance in the catchment between 1951 and 1967. Due to their asynchronous occurrence, eutrophication and catchment disturbance could not be the major drivers of hypolimnetic deoxygenation. Interestingly, climate records indicated that the local ambient temperature exhibited similar trends to that of hypolimnetic deoxygenation, including markedly higher temperatures after 1994. This implies that prolonged water column stratification due to climate warming could have promoted hypolimnetic deoxygenation by limiting oxygen exchange between the upper and lower water columns. In comparing the results with those of Lake Erhai, another monomictic lake with contrasting morphology and hydrology characteristics, we found that the low surface area/depth ratio and long water residence time in Lake Fuxian enable dissolved oxygen of the lake hypolimnion vulnerable to temperature over the lake.

Abstract Background Numerous studies have manifested long noncoding RNAs (lncRNAs) as biomarkers to determine the prognosis of multiple myeloma (MM) patients. Nevertheless, the prognostic role of lncRNAs in MM is still ambiguous. Herein, we performed a meta‐analysis to evaluate the predictive value of aberrantly expressed lncRNAs in MM. Methods A systemic literature search was performed in PubMed, EMBASE, Cochrane, and Web of Science databases until October 9, 2021, and the protocol was registered in the PROSPERO database (CRD42021284364). Our study extracted the hazard ratios (HRs) and 95% confidence intervals (CIs) of overall survival (OS), progression‐free survival (PFS), or event‐free survival (EFS). Begg's and Egger's tests were employed to correct publication bias. Result Twenty‐six individual studies containing 3501 MM patients were enrolled in this study. The results showed that aberrant expression of lncRNAs was associated with poor OS and PFS of MM patients. The pooled HRs for univariate OS and PFS were 1.48 (95% CI = 1.17–1.88, p

Event logs of business processes provide a valuable starting point for process mining, and high-quality event logs can significantly enhance the quality of process mining. However, event logs often contain a substantial amount of sensitive and personal information. Therefore, the release of event logs should prioritize the model’s quality while minimizing the risk of privacy exposure. Specifically, quantifying performance indicators between the original event logs and the released ones enables the operational goals. To date, privacy benefit and utility loss are two main target performance indicators, especially from the perspective of structural similarity comparison of mined process models. To the best of our knowledge, no study aims to measure the privacy-preserving performance indicators from the point of behavior differentiation between the original event logs and released ones. In this paper, we propose an approach to quantify the behavior differentiation between the original event logs and the corresponding released ones. Specifically, an approach of event log release mechanism that effectively combines behavior privacy gain and behavior utility loss is presented in this paper. Firstly, we discuss challenges in scenarios where event data is released without privacy preservation, and describe potential attacks that could occur when third-party businesses perform process mining techniques. Based on these potential attacks, we present a behavior differentiation-based event log release mechanism named PLI-Assess to combat these threats. Finally, we conduct experiments on four groups of practical event logs for comparisons with the baseline methods.The experimental results suggest feasibility of privacy-utility trade-offs.

It is a fact that plenty of process variants are derived from the same base model in practical applications, and the main reasons include inevitable software maintenance and adaptability of process models. This fact raises a question that many process variants are with high similarity degree, and can not be differentiated from each other under the state-of-the-art similarity measurements. In order to differentiate similar-but-different process variants, we propose an approach of process variants’ behavior differentiation in this paper. The method analyzes the complex structures of process variants with invisible tasks, and utilizes task execution relationship to construct an integrated similarity measurement, which extends the existing similarity measurement capacity in terms of dealing with invisible tasks. It is proved that the proposed task execution relationship can capture the dominate features of similar-but-different process variant including invisible tasks, which means that as long as the task execution behaviors of two process variants are different, the corresponding behavior matrices must be different. Furthermore, a set of experiments are carried out, in order to evaluate the properties of effectiveness, semantic uniqueness expression and correctness of the proposed method. Meanwhile, the experimental results give evidences that the proposed method outperforms the existing model similarity measurements in accuracy.

ObjectiveTo analyze the surveillance results of influenza-like illness （ILI） cases in a sentinel hospital in Nantong from 2014 to 2021 and to understand the seasonal and etiological characteristics of influenza.MethodsThe routine ILI case information in a sentinel hospital in Nantong from 2014 to 2021 was collected from China Influenza Surveillance Information system， and the surveillance results were described and analyzed.ResultsA total of 64 110 ILI cases were reported in the hospital from 2014 to 2021， and accounted for 3.93% of outpatient and emergency visits. ILI is mainly concentrated in children under 15 years old， accounting for 55.39% of the total ILI cases. ILI% roughly showed two epidemic peaks in spring and winter. A total of 8 639 nasopharyngeal swab samples of ILI cases were collected during 2014‒2021， of which 838 were influenza positive， with an average positive rate of 10.01% （3.27%‒16.75%）. The epidemic strains varied from year to year and in different age groups. In 2014 and 2015， H3N2 influenza was the epidemic strain； in 2016 and 2017， H3N2 influenza and B-Victoria influenza were the co-epidemic strains； in 2018， H1N1 influenza was the epidemic strain； and in 2019‒2021， B Victoria influenza was the epidemic strain. The epidemic strain in 0‒4 years old， 15‒24 years old， 25‒59 years old and >60 years old was H3N2， and the epidemic strain in 5‒14 years old was B Victoria. There was a significant difference in the positive rate between different gender groups （χ2=7.364， P=0.006）.ConclusionInfluenza often occurs in winter and spring in a sentinel hospital in Nantong. Influenza subtypes are prevalent alternately in different age groups and different years. We need to closely monitor the etiology of influenza virus in Nantong to provide a basis for influenza prevention and control.

Abstract Background Erectile dysfunction (ED), as one of the most prevalent consequences in male diabetic patients, has a serious impact on men's physical and mental health, and the treatment effect of diabetic mellitus erectile dysfunction (DMED) is often worse. Therefore, the development of a novel therapeutic approach is urgent. As stem cells with high differentiation potential, human umbilical cord mesenchymal stem cells (HUCMSCs) have been widely used in the treatment of diseases in other systems, and are expected to be a promising strategy for the treatment of DMED. In this study, we investigated the role of HUCMSCs in managing erectile function in rat models of type 1 diabetes mellitus (T1DM) and type 2 diabetes mellitus (T2DM) and compared the effects of two different injection methods. Methods T1DM and T2DM ED rats were given labelled HUCMSCs by corpus cavernosum injection and tail vein injection, respectively. ICP and MAP were monitored simultaneously by electrical stimulation four weeks after injection to indicate the erectile function of rats. To track the development and colonisation capabilities of stem cells, we performed EdU assay with penile tissue. The histological changes of the penis were observed by hematoxylin–eosin staining, and Masson’s trichrome staining was conducted to evaluate the smooth muscle content and the degree of fibrosis in the rat penis. Then, we employed specific kits to measure the level of NO, cGMP, MDA, SOD and Fe in penis. Electron transmission microscopy was implemented to observe morphology of mitochondria. Besides, western blot and immunofluorescence staining were performed to demonstrate the expression of ferroptosis-related genes. Results We found that HUCMSCs improved erectile function in T1DM and T2DM ED rats, with no difference in efficacy between corpus cavernosum injection and tail vein injection. The EdU assay revealed that only a tiny percentage of HUCMSCs colonised the corpus cavernosum, while smooth muscle in the penis expanded and collagen decreased following HUCMSC injection. Moreover, the levels of oxidative stress in the penis of the rats given HUCMSCs were dramatically reduced, as was the tissue iron content. HUCMSCs normalised mitochondrial morphology within corpus cavernosum smooth muscle cells (CCSMCs), which were characteristically altered by high glucose. Furthermore, the expression of ferroptosis inhibitory genes SLC7A11 and GPX4 was obviously elevated in CCSMCs after stem cell management, but the abundances of ACSL4, LPCAT3 and ALOX15 showed the polar opposite tendency. Conclusions HUCMSCs can effectively and safely alleviate erectile dysfunction in T1DM and T2DM ED rats, while restoring erectile function by attenuating diabetes-induced ferroptosis in CCSMCs. Additionally, this study provides significant evidence for the development of HUCMSCs as a viable therapeutic strategy for DMED.

BackgroundUrothelial carcinoma of the bladder (UCB) is the most prevalent malignant tumor of the urinary system worldwide, which has a significant recurrence rate despite multiple treatment options available. As a unique and novel copper-dependent programmed cell death mechanism, the comprehensive impact of cuproptosis on the tumor immune microenvironment, clinicopathological characteristics and the prognosis of patients remains largely unclear.MethodsA total of 568 UCB samples were thoroughly examined for cuproptosis patterns using data downloaded from TCGA and GEO, based on 10 cuproptosis-related genes reported previously. Then, the univariate COX regression analysis was performed on the genes that differed across the various patterns. To measure individual cuproptosis pattern, a cuproptosis score system was constructed using a principal component analysis algorithm. To validate the scoring system, immunohistochemical staining was performed on tumor tissues with different pathological grades, and experiments in vitro were conducted about the differentially expressed genes related to prognosis. Finally, the capacity of scoring system to predict the response to immunotherapy was verified by using data from IMvigor 210 cohort.ResultsFour unique cuproptosis clusters and two gene clusters were finally found by the investigation. The clinical features and prognosis of patients, as well as the mRNA transcriptome, pathway enrichment, and immune cell infiltration in TME, varied dramatically between various cuproptosis clusters and gene clusters. To identify individual cuproptosis patterns in UCB patients, we also established a cuproptosis scoring system. After validation with multiple methods, it was indicated that the score system could predict the prognosis of UCB patients and was significantly connected to clinical features such TNM category, tumor grade, molecular type and ultimate survival status. The clinical outcomes of UCB patients were predicted effectively according to the tumor mutation burden in conjunction with the scoring system. Furthermore, we found that the cuproptosis score had a significant correlation with the response to immunotherapy and the sensitivity to chemotherapy.ConclusionThis study revealed the potential impact of cuproptosis on the UCB tumor immune microenvironment and clinical pathological characteristics. The cuproptosis score system could effectively predict the prognosis of patients and the response to chemotherapy and immunotherapy.

Glucose management for people with type 2 diabetes mellitus is essential but challenging due to the multi-factored and chronic disease nature of diabetes. To control glucose levels in a safe range and lessen abnormal glucose variability efficiently and economically, an intelligent prediction of glucose is demanding. A glucose trajectory prediction system based on subcutaneous interstitial continuous glucose monitoring data and deep learning models for ensuing glucose trajectory was constructed, followed by the application of personalised prediction models on one participant with type 2 diabetes in a community. The predictive accuracy was then assessed by RMSE (root mean square error) using blood glucose data. Changes in glycaemic parameters of the participant before and after model intervention were also compared to examine the efficacy of this intelligence-aided health care. Individual Recurrent Neural Network model was developed on glucose data, with an average daily RMSE of 1.59 mmol/L in the application segment. In terms of the glucose variation, the mean glucose decreased by 0.66 mmol/L, and HBGI dropped from 12.99 × 102 to 9.17 × 102. However, the participant also had increased stress, especially in eating and social support. Our research presented a personalised care system for people with diabetes based on deep learning. The intelligence-aided health management system is promising to enhance the outcome of diabetic patients, but further research is also necessary to decrease stress in the intelligence-aided health management and investigate the stress impacts on diabetic patients.

Abstract Inorganic nitrate production is critical in atmospheric chemistry that reflects the oxidation capacity and the acidity of the atmosphere. Here we use the oxygen anomaly of nitrate (Δ17O( $$\rm{NO}_{3}^{-}$$ NO 3 − )) in high-time-resolved (3 h) aerosols to explore the chemical mechanisms of nitrate evolution in fine particles during the winter in Nanjing, a megacity of China. The continuous Δ17O( $$\rm{NO}_{3}^{-}$$ NO 3 − ) observation suggested the dominance of nocturnal chemistry (NO3 + HC/H2O and N2O5 + H2O/Cl−) in nitrate formation in the wintertime. Significant diurnal variations of nitrate formation pathways were found. The contribution of nocturnal chemistry increased at night and peaked (72%) at midnight. Particularly, nocturnal pathways became more important for the formation of nitrate in the process of air pollution aggravation. In contrast, the contribution of daytime chemistry (NO2 + OH/H2O) increased with the sunrise and showed a highest fraction (48%) around noon. The hydrolysis of N2O5 on particle surfaces played an important role in the daytime nitrate production on haze days. In addition, the reaction of NO2 with OH radicals was found to dominate the nitrate production after nitrate chemistry was reset by the precipitation events. These results suggest the importance of high-time-resolved observations of Δ17O( $$\rm{NO}_{3}^{-}$$ NO 3 − ) for exploring dynamic variations in reactive nitrogen chemistry.

Abstract The piezoelectric nanopositioning platform requires extremely accurate tracking during the task, while the model uncertainty caused by load variations requires strong robustness of the system. The high accuracy and robustness in the control design are coupled to each other, making it difficult to achieve both optimally at the same time. In addition, the system itself has a weakly damped resonant mode, which makes it extremely difficult to control the piezoelectric nanopositioning platform while suppressing the inherent resonance of the system as well as meeting the requirements for robustness and high accuracy. For the multi-performance integrated control problem of piezoelectric nanopositioning platform, this paper gives two kinds of control designs (integral resonance control (IRC) and H∞ control) satisfying accuracy requirements and robustness, and carries out simulation study and comparative analysis with positive position feedback control (PPF). Simulation results show that the H∞ control strategy given in this paper has the smallest tracking error compared to PPF and IRC under 5, 10 and 20 Hz input grating scan signals, though it has a higher order, with better robustness to mechanical load variations and high frequency signal perturbations in the 0–1000 g load range.

Erectile dysfunction (ED), as a common male disease, can seriously reduce the life quality of men and their partners. With the improvement of human living standards, ED is considered to be an important health issue that plagues men. However, it is difficult for existing therapeutic approaches to meet the needs of all patients, so it is necessary to develop novel treatment strategies. Exosomes, as a class of vesicles secreted by cells with bilayer membrane structure, are involved in various physiological and pathological processes in human body and considered to have great therapeutic potentials. This review summarizes the recent advances on exosome therapy with animal models of ED, and proposes the prospect of future research in order to provide a basis for clinical trials and clinical translation.

Abstract Background The reliability of Criteria for Assessing Prescription Quality in Chinese Hospitals (CAPQCH) has never been rigorously verified. This study was designed to verify the reliability of the CAPQCH among pharmacists in China. Methods Fourteen pharmacists, 5 from hospitals and 9 from the communities were recruited. We randomly selected 200 prescriptions, and made the testing prescriptions including appropriate and inappropriate testing prescriptions. Pharmacists assessed these testing prescriptions according to criteria in CAPQCH. Three test sets (Set 1, Set 2, and Set 3) were evaluated at 6-month intervals. Before administration of Set 3, pharmacists were informed that achievement on Set 3 would be reflected in their performance appraisal. We also evaluated the performance based on prescription comments before and after combining several confusing criteria. Cohen’s Kappa statistic, Fleiss’ Kappa statistic, and accuracy were employed to evaluate reliability among pharmacists. Results Median values of Cohen’s Kappa were 0.61 in Set 1, 0.66 in Set 2, and 0.80 in Set 3; reliability is thus substantial. Our data indicate no significant differences between Set 1 and Set 2, whereas Set 3 indicates significantly improved performance. Moreover, combinations of confusing criteria contributed little to improvement of performance in prescription comments. Conclusion Our results verified the reliability of CAPQCH application by working pharmacists. Adding performance based on prescription comments to personal appraisals was effective in improving the quality of prescription comments. These findings may be useful when future modification of the CAPQCH is considered. Moreover, this study contributes to improving the understanding of the prescription assessment situation in China.

On a daily basis, a wide range of materials including inorganic nanoparticles (NPs) inadvertently find their way into the environment. Meanwhile, intentionally used NPs, such as the new generation of nanofertilizers (NFs) are designed to enhance agronomic production. However, their physicochemical properties and not-so-well understood effects raise potential risks to the plant reproductive cycle, specifically pollen development, a subject largely absent in academic research. Even slight contamination, deformation, or aberration of pollen could have enormous impacts on the ecosystem. Thus, our objective was to evaluate the influence of various metal-based NPs on sunflower pollen morphology and its yield. Nano-formulations were applied during the 2019–2021 agronomic seasons on two sunflower hybrids, Neostar and Edison, in Dolná Malanta, near Nitra, Slovak Republic. Pollen morphology findings indicated that conventional ZnSO4 had the most positive impact on the size of pollen grains compared to ZnO-NPs, Fe3O4-NPs, and the NP-free control. Gold-NPs on SiO2 mesoporous silica (AuSi-NPs) showed a statistically insignificant impact, while the use of TiO2-NPs in agriculture remained a topic of debate. Surprisingly, pollen characteristics did not fully correspond to crop yields. Despite causing a reduction in pollen grain size, the TiO2-NPs consistently showed the highest yield compared to other variants. Employing low concentrations of NFs did not notably alter pollen morphology, reinforcing our commitment to eco-friendly, precise, and sustainable agriculture.

To minimize the eutrophication pressure along the Gulf of Mexico or reduce the size of the hypoxic zone in the Gulf of Mexico, it is important to understand the underlying temporal and spatial variations and correlations in excess nutrient loads, which are strongly associated with the formation of hypoxia. This study’s objective was to reveal and visualize structures in high-dimensional datasets of nutrient yield distributions throughout the Mississippi/Atchafalaya River Basin (MARB). For this purpose, the annual mean nutrient concentrations were collected from thirty-three US Geological Survey (USGS) water stations scattered in the upper and lower MARB from 1996 to 2020. Eight surface water quality indicators were selected to make comparisons among water stations along the MARB over the past two decades. Principal component analysis (PCA) was used to comprehensively evaluate the nutrient yields across thirty-three USGS monitoring stations and identify the major contributing nutrient loads. The results showed that all samples could be analyzed using two main components, which accounted for 81.6% of the total variance. The PCA results showed that yields of orthophosphate (OP), silica (SI), nitrate–nitrites (NO3-NO2), and total suspended sediment (TSS) are major contributors to nutrient yields. It also showed that land-planted crops, density of population, domestic and industrial discharges, and precipitation are fundamental causes of excess nutrient loads in MARB. These factors are of great significance for the excess nutrient load management and pollution control of the Mississippi River. It was found that the average nutrient yields were stable within the sub-MARB area, but the large nitrogen yields in the upper MARB and the large phosphorus yields in the lower MARB were of great concern. t-distributed stochastic neighbor embedding (t-SNE) revealed interesting nonlinear and local structures in nutrient yield distributions. Clustering analysis (CA) showed the detailed development of similarities in the nutrient yield distribution. Moreover, PCA, t-SNE, and CA showed consistent clustering results. This study demonstrated that the integration of dimension reduction techniques, PCA, and t-SNE with CA techniques in machine learning are effective tools for the visualization of the structures of the correlations in high-dimensional datasets of nutrient yields and provide a comprehensive understanding of the correlations in the distributions of nutrient loads across the MARB.

Background: The aim of this study was to evaluate the long-term sequelae and cognitive profiles resulting from severe hand, foot, and mouth disease (HFMD) with central nervous system (CNS) involvement. Methods: 294 HFMD cases were included in a retrospective follow-up study. Physical examinations were conducted. The Chinese Wechsler Preschool and Primary Scale of Intelligence, Fourth Edition (WPPSI-IV) was used to assess intelligence. Results: 58 mild HFMD cases and 99 severe HFMD cases with mild CNS involvement did not present any neurological sequelae. In comparison, the sequelae incidence for severe HFMD with more severe CNS complications was 50.0%. The proportion of full-scale intelligence quotient (FSIQ) impairment was 45.0%. In the 2:6–3:11 age group, severe HFMD with more severe CNS complications and lower maternal education level were risk factors for verbal comprehension disorder. Urban–rural residence and lower paternal education level were risk factors for FSIQ disorder. Furthermore, in the 4:0–6:11 age group, severe HFMD with more severe CNS complication was a risk factor for visual spatial disorder and fluid reasoning disorder. Lower paternal education level was a risk factor for FSIQ disorder. Conclusion: Early assessment and intervention among severe HFMD patients with more severe CNS involvement at a very young age will prove beneficial for their future performance.

Background: Enhanced recovery after surgery (ERAS) has received increasing attention. Preoperative oral carbohydrate and postoperative early oral feeding (POC-PEOF) as the basic nutrition administration in the ERAS program suffers from low adherence. The role and benefits of administering POC-PEOF in elderly patients with hepatocellular carcinoma (HCC) are unclear. Therefore, the randomized controlled trial evaluated the effects of POC-PEOF in elderly patients with HCC undergoing hepatectomy with inflammation and patient self-reported symptom burden compared with the corresponding outcomes of traditional fasting protocols. Methods: Elderly patients with HCC (n = 126) were randomly assigned to two groups using the sealed envelope technique. Sixty-three patients were included in the intervention (POC-PEOF) group and received POC-PEOF administration, whereas the 63 patients in the control (FAST) group underwent conventional fasting. Acute-phase inflammation markers, patient self-reported symptom burdens, and postoperative outcomes were compared between the two groups. Results: The average age was 69.60 ± 5.00 years in the POC-PEOF group and 70.44 ± 6.15 years in the FAST group. Compared to prolonged fasting, POC-PEOF achieved significant positive results, including lower overall levels of inflammatory response mediators (CRP, IL-6) on postoperative day (POD) 1, POD 3, and POD 5 (P

Abstract Background Proteases catalyze the hydrolysis of peptide bonds of proteins, thereby improving dietary protein digestibility, nutrient availability, as well as flavor and texture of fermented food and feed products. The lactobacilli Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) and Pediococcus acidilactici are widely used in food and feed fermentations due to their broad metabolic capabilities and safe use. However, extracellular protease activity in these two species is low. Here, we optimized protease expression and secretion in L. plantarum and P. acidilactici via a genetic engineering strategy. Results To this end, we first developed a versatile and stable plasmid, pUC256E, which can propagate in both L. plantarum and P. acidilactici. We then confirmed expression and secretion of protease PepG1 as a functional enzyme in both strains with the aid of the previously described L. plantarum-derived signal peptide LP_0373. To further increase secretion of PepG1, we carried out a genome-wide experimental screening of signal peptide functionality. A total of 155 predicted signal peptides originating from L. plantarum and 110 predicted signal peptides from P. acidilactici were expressed and screened for extracellular proteolytic activity in the two different strains, respectively. We identified 12 L. plantarum signal peptides and eight P. acidilactici signal peptides that resulted in improved yield of secreted PepG1. No significant correlation was found between signal peptide sequence properties and its performance with PepG1. Conclusion The vector developed here provides a powerful tool for rapid experimental screening of signal peptides in both L. plantarum and P. acidilactici. Moreover, the set of novel signal peptides identified was widely distributed across strains of the same species and even across some closely related species. This indicates their potential applicability also for the secretion of other proteins of interest in other L. plantarum or P. acidilactici host strains. Our findings demonstrate that screening a library of homologous signal peptides is an attractive strategy to identify the optimal signal peptide for the target protein, resulting in improved protein export.

The inherent weakly damped resonant modes of the piezoelectric nanopositioning platform and the presence of model uncertainty seriously affect the performance of the system. A structured H∞ design is used in this paper to solve the accuracy and robustness problems respectively using a two-loop control structure. The multiple performance requirements of the system are constituted into an H∞ optimization matrix containing multi-dimensional performance diagonal decoupling outputs, and an inner damping controller d is set according to the damping of the resonant modes; the second-order robust feedback controller is preset in the inner loop to improve the robustness of the system; the tracking controller is connected in series in the outer loop to achieve high accuracy scanning; finally, the structured H∞ controller is designed to meet the multiple performance requirements. To verify the effectiveness of the proposed structured H∞ control, simulation comparison experiments are done with the integral resonant control (IRC) and H∞ controller. The results demonstrate that the designed structured H∞ controller achieves higher tracking accuracy compared to the IRC and H∞ controllers under grating input signals of 5, 10, and 20 Hz. Moreover, it has good robustness under 600g and 1000g loads and high frequency disturbances close to the resonant frequency of the system, meeting multiple performance requirements. Compared with the traditional H∞ control, yet with lower complexity and transparency, which is more suitable for engineering practice applications.

The fungus Curvularia tsudae can survive in environments that are extremely contaminated by heavy metals; however, the underlying molecular mechanisms of heavy metal tolerance are not clear. In this study, we determined the effects of lead (Pb) stress on the growth of C. tsudae and used RNA-Seq to identify significant genes and biological processes involved. The present study showed that C. tsudae had an outstanding resistant capacity to Pb stress and could survive at a concentration of 1600 mg L−1 Pb. Although an obvious inhibition on the growth was observed, the fungus exhibited tolerance as it continued to grow at a Pb concentration of 1600 mg L−1 for seven days. A total of 9997 (9020 up and 977 down) differentially expressed genes (DEGs) were detected in the mycelium of C. tsudae at Pb free (0 mg L−1) and Pb stressed samples. Pathway enrichment analysis identified several biological processes for managing Pb stress. Genes involved in carbohydrate metabolism tended to be modulated in response to Pb stress, while amino acids and the lipid metabolism would also be induced by Pb stress, and up-regulated genes involved in antioxidant substances and ABC transporters may be committed to high Pb tolerance. Our study contributes to the current literature on C. tsudae response to Pb stress and provides a useful reference for fungi as bioremediators in heavy metal-contaminated environments.

Dynamic regulation of transcription is crucial for the cellular responses to various environmental or developmental cues. Gdown1 is a ubiquitously expressed, RNA polymerase II (Pol II) interacting protein, essential for the embryonic development of metazoan. It tightly binds Pol II in vitro and competitively blocks the binding of TFIIF and possibly other transcriptional regulatory factors, yet its cellular functions and regulatory circuits remain unclear. Here, we show that human GDOWN1 strictly localizes in the cytoplasm of various types of somatic cells and exhibits a potent resistance to the imposed driving force for its nuclear localization. Combined with the genetic and microscope-based approaches, two types of the functionally coupled and evolutionally conserved localization regulatory motifs are identified, including the CRM1-dependent nucleus export signal (NES) and a novel Cytoplasmic Anchoring Signal (CAS) that mediates its retention outside of the nuclear pore complexes (NPC). Mutagenesis of CAS alleviates GDOWN1’s cytoplasmic retention, thus unlocks its nucleocytoplasmic shuttling properties, and the increased nuclear import and accumulation of GDOWN1 results in a drastic reduction of both Pol II and its associated global transcription levels. Importantly, the nuclear translocation of GDOWN1 occurs in response to the oxidative stresses, and the ablation of GDOWN1 significantly weakens the cellular tolerance. Collectively, our work uncovers the molecular basis of GDOWN1’s subcellular localization and a novel cellular strategy of modulating global transcription and stress-adaptation via controlling the nuclear translocation of GDOWN1.

Nyctocalos is a genus of flowering lianas belonging to the family Bignoniaceae, and occurring from South-Central China to Malesia. In this study, we assembled the first complete chloroplast genome of N. pinnatum. The total length of the chloroplast genome is 159591 bp, with a GC content of 38.04%, which includes a pair of inverted repeats of 30,480 bp, a small single-copy region of 12,774 bp and a large single-copy region of 85,857 bp. The chloroplast genome contains 135 genes, consisting of 89 protein-coding genes, 38 transfer RNAs, and 8 ribosomal RNAs. We constructed a phylogenomic tree with representative chloroplast genomes from Bignoniaceae. N. pinnatum is revealed to be sister to Oroxylum in the tribe Oroxyleae, with a high bootstrap support. This is the first chloroplast genome assembled in Nyctocalos, and it provides essential information for further ecology and evolutionary studies in this genus and Bignoniaceae.

PCSK9 negatively regulates low-density lipoprotein receptor (LDLR) abundance on the cell surface, leading to decreased hepatic clearance of LDL particles and increased levels of plasma cholesterol. We previously identified SURF4 as a cargo receptor that facilitates PCSK9 secretion in HEK293T cells (Emmer et al., 2018). Here, we generated hepatic SURF4-deficient mice (Surf4fl/fl Alb-Cre+) to investigate the physiologic role of SURF4 in vivo. Surf4fl/fl Alb-Cre+ mice exhibited normal viability, gross development, and fertility. Plasma PCSK9 levels were reduced by ~60% in Surf4fl/fl Alb-Cre+ mice, with a corresponding ~50% increase in steady state LDLR protein abundance in the liver, consistent with SURF4 functioning as a cargo receptor for PCSK9. Surprisingly, these mice exhibited a marked reduction in plasma cholesterol and triglyceride levels out of proportion to the partial increase in hepatic LDLR abundance. Detailed characterization of lipoprotein metabolism in these mice instead revealed a severe defect in hepatic lipoprotein secretion, consistent with prior reports of SURF4 also promoting the secretion of apolipoprotein B (APOB). Despite a small increase in liver mass and lipid content, histologic evaluation revealed no evidence of steatohepatitis or fibrosis in Surf4fl/fl Alb-Cre+ mice. Acute depletion of hepatic SURF4 by CRISPR/Cas9 or liver-targeted siRNA in adult mice confirms these findings. Together, these data support the physiologic significance of SURF4 in the hepatic secretion of PCSK9 and APOB-containing lipoproteins and its potential as a therapeutic target in atherosclerotic cardiovascular diseases.

Bovine mastitis caused by Streptococcus dysgalactiae (S. dysgalactiae) is usually treated with antibiotics, which may potentially increase drug resistance as the abuse. NZ2114, a variant of fungal defensin plectasin, displayed a potent antibacterial activity against S. dysgalactiae. The inhibition/eradication effect of the antimicrobial peptide NZ2114 on the early/mature biofilm of S. dysgalactiae CVCC 3938 was evaluated, as well as the elimination of bacteria in mature biofilms. In this study, NZ2114 displayed potent antibacterial activity against S. dysgalactiae CVCC 3938 and three clinical isolated S. dysgalactiae strains (0.11-0.45 μM). The early biofilm inhibition of S. dysgalactiae CVCC 3938 was 55.5–85.9% after treatment with NZ2114 at concentrations of 1–16 × MIC, which was better than that of vancomycin at the same concentration. The mature biofilm eradication rate was up to 92.7–97.6% with the increasing concentration (2–16 × MIC) of NZ2114, and the eradication rate did not change significantly with further increase of NZ2114 concentration, while the biofilm eradication rate of vancomycin-treated group at the same concentration remained at 92.5%. NZ2114 reduced the number of persister bacteria in biofilm. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) further demonstrated that NZ2114 could effectively reduce the biofilm thickness and bacterial number of S. dysgalactiae CVCC 3938. In vivo therapeutic effect of NZ2114 on murine mastitis model showed that NZ2114 was better than vancomycin in alleviating mammary gland inflammation by regulating cytokines production, inhibiting bacterial proliferation, and reducing the number of mammary gland bacteria. These data suggested that NZ2114 is a potential peptide candidate for the treatment of mastitis.

In this work, a fluorophores-assisted excitation/emission matrix (EEM) fluorescence method was proposed to trace the origin of lily in the Chinese market. There are few active components in lilies that have fluorescent signals, and too few characteristic variables may lead to unsatisfactory accuracy in the subsequent classification. Therefore, three fluorophores, 2-Aminoethyl diphenylborinate (DPBA), o-Phthalaldehyde (OPA) and Rhodamine B (RB), were used to enrich the information of the fluorescent fingerprint of lily, which can improve the classification accuracy. The lily samples were characterized by using EEM fluorescence coupled with the alternating trilinear decomposition (ATLD) algorithm, which was able to extract information of various fluorophores in lily samples. Two chemical pattern recognition methods, principal component analysis-linear discriminant analysis (PCA-LDA) and partial least squares-discrimination analysis (PLS-DA), were used to model and trace the origin of different lilies. When the fluorophores were added, the accuracy of the test set and prediction set obtained by the classification model increased from 71.4% to 92.9% and 66.7% to 100%, respectively. The proposed method combined fluorophores-assisted EEM fluorescence with multi-way chemometric methods to extract comprehensive information on the samples, which provided a potential method for the origin traceability of traditional Chinese medicine.