Relish-mediated C2H2 zinc finger protein IMZF restores Drosophila immune homeostasis via inhibiting the transcription of Imd/Tak1.

The dysregulation of intensity and duration in innate immunity can result in detrimental effects on the body, emphasizing the crucial need for precise regulation. However, the intricate and accurate nature of innate immunity implies the existence of numerous undiscovered innate immunomodulators, particularly transcription factors. In this study, we have identified a Drosophila C2H2 zinc finger protein CG18262, named Immune-mediated Zinc Finger protein (IMZF), capable of suppressing immune responses of Imd pathway. Mechanistically, IMZF serves as a transcription factor that represses the expression of Imd and Tak1. Intriguingly, our findings also reveal that Relish, an NF-κB transcription factor, positively regulates the expression of IMZF, consequently inhibiting the activation of Imd and Tak1 to prevent an exaggerated immune response. Additionally, we have elucidated the pivotal role played by the Relish-IMZF-Imd/Tak1 axis in restoring immune homeostasis of Drosophila Imd pathway. In summary, our findings not only unveil a novel C2H2 zinc finger immunoregulatory transcription factor, IMZF, along with its specific mechanism of immune regulation, but also shed light on the dual functionality of Relish in different stages of the immune response by modulating distinct effectors. This discovery provides new insights and enlightenment into the complex regulation of Drosophila innate immunity.

NF-κB factors cooperate with Su(Hw)/E4F1 to balance Drosophila/human immune responses via modulating dynamic expression of miR-210.

MicroRNAs (miRNAs) play crucial regulatory roles in controlling immune responses, but their dynamic expression mechanisms are poorly understood. Here, we firstly confirm that the conserved miRNA miR-210 negatively regulates innate immune responses of Drosophila and human via targeting Toll and TLR6, respectively. Secondly, our findings demonstrate that the expression of miR-210 is dynamically regulated by NF-κB factor Dorsal in immune response of Drosophila Toll pathway. Thirdly, we find that Dorsal-mediated transcriptional inhibition of miR-210 is dependent on the transcriptional repressor Su(Hw). Mechanistically, Dorsal interacts with Su(Hw) to modulate cooperatively the dynamic expression of miR-210 in a time- and dose-dependent manner, thereby controlling the strength of Drosophila Toll immune response and maintaining immune homeostasis. Fourthly, we reveal a similar mechanism in human cells, where NF-κB/RelA cooperates with E4F1 to regulate the dynamic expression of hsa-miR-210 in the TLR immune response. Overall, our study reveals a conservative regulatory mechanism that maintains animal innate immune homeostasis and provides new insights into the dynamic regulation of miRNA expression in immune response.

The identification of regulatory ceRNA network involved in Drosophila Toll immune responses.

Non-coding RNAs play important roles in the innate immunity of Drosophila, with various lncRNAs and miRNAs identified to maintain Drosophila innate immune homeostasis by regulating protein functions. However, it remains unclear whether interactions between lncRNAs and miRNAs give rise to a ceRNA network. In our previous study, we observed the highest differential expression levels of lncRNA-CR11538, lncRNA-CR33942, and lncRNA-CR46018 in wild-type flies after Gram-positive bacterial infection, prompting us to investigate their role in the regulation of Drosophila Toll immune response through RNA-seq analysis. Herein, our comprehensive bioinformatics analysis revealed that lncRNA-CR11538, lncRNA-CR33942, and lncRNA-CR46018 are involved in defense mechanisms and stimulus response. Moreover, lncRNA-CR11538 and lncRNA-CR46018 can also participate in the metabolic recovery processes following Gram-positive bacterial infection. Subsequently, we employed GSEA screening and RT-qPCR to identify seven miRNAs (miR-957, miR-1015, miR-982, miR-993, miR-1007, miR-193, and miR-978) that may be regulated by these three lncRNAs. Furthermore, we predicted the potential target genes in the Toll signaling pathway for these miRNAs and their interaction with the three lncRNAs using TargetScan and miRanda software and preliminary verification. As a result, we established a potential ceRNA regulatory network for Toll immune responses in Drosophila, comprising three lncRNAs and seven miRNAs. This study provides evidence of a ceRNA regulatory network in Drosophila Toll immune responses and offers novel insights into understanding the regulatory networks involved in the innate immunity of other animals.

Synchronized delivery of dual-drugs for potentiating combination chemotherapy based on smart triple-responsive polymeric micelles.

Here, we combined reversible addition-fragmentation chain transfer (RAFT) polymerization and amide coupling reaction to develop a novel drug-polymer conjugate using poly(AMA-co-IMMA)-b-poly(OEGMA) (termed as PAIPO) as nanocarriers. In order to enhance cellular uptake and obtain subsequent endo/lysosomal escape capacity, the dual-drugs-conjugated prodrug was then coupled with 2,3-dimethylmaleimide (DA) moieties and implanted with imidazolyl groups, respectively. Paclitaxel (PTX) was conjugated to PAIPO via 3,3'-dithiodipropionic acid (DPA) to construct a GSH-responsive moiety, while doxorubicin (DOX) was conjugated to PAIPO via 4-formyl benzoic acid to construct a pH-responsive moiety, which synergistically enabled a synchronized and precise drug delivery. The micelles self-assembled from DOX/PTX@PAIPODA showed an ideal average diameter (163.2-178.3 nm), contributing to passive targeting by the EPR effect. Moreover, a switch of the surface Zeta potential of micelles from steady negatively charged (- 9.74 ± 0.54 mV) at pH 7.4 to positively charged (+ 6.33 ± 1.25 mV) at pH 6.5, facilitated the long blood circulation and cellular endocytosis of micelles, respectively. More importantly, in vitro studies confirmed that DAM(DOXn/PTX) exhibited a strong synergism against tumor cells, and under slightly acidic conditions (pH 6.5), the combination index (CI) values for DAM(DOX1/PTX) on HeLa and Skov-3 cells were estimated to be 0.47 and 0.49 (previous to be 0.50 and 0.56 at pH 7.4), respectively. And in vivo results showed effective tumor accumulation potential, remarkable biosafety, and biocompatibility. Combined, such synchronized delivery approach based on multi-responsive micelles might potentiate the efficacy of combination chemotherapy in clinical cancer treatment.

Prediction of the Short-Term Risk of New-Onset Renal Dysfunction in Patients with Type 2 Diabetes: A Longitudinal Observational Study.

Background: Studies in the past decade have reported many novel biomarkers for predicting the new-onset or progression risk of renal dysfunction in patients with type 2 diabetes (T2D) based on the genomic, metabolomic, and proteomic technologies. These novel predictive markers, however, are difficult to be widely used in clinical practice over the short term due to their high technology content, instability, and high cost. This study was aimed at evaluating the associations of clinical features and six traditional renal markers with the short-term risk of new-onset renal dysfunction in patients with T2D. Methods: This study involved 213 participants with T2D and normal renal function at baseline. The baseline levels of the albumin-to-creatinine ratio (ACR), estimated glomerular filtration rate (eGFR), alpha-1-microglobulin-to-creatinine ratio (A1MCR), neutrophil gelatinase-associated lipocalin-to-creatinine ratio, transferrin-to-creatinine ratio (UTRF/Cr), and retinol-binding protein-to-creatinine ratio (URBP/Cr) were analyzed. Multivariate logistic models were established and validated. Results: During the two-year follow-up period, 23.01% participants progressed to renal dysfunction. The basal levels of ACR, A1MCR, UTRF/Cr, and URBP/Cr were the independent risk factors of new-onset renal dysfunction (P < 0.05). Several logistic models incorporating clinical characteristics and these renal markers were constructed for predicting the short-term risk of new-onset renal dysfunction. Comparatively, the model including age, glycated hemoglobin (HbA1c), hypertension, ACR, A1MCR, UTRF/Cr, and URBP/Cr levels at baseline had the highest potential (C - index = 0.785, P < 0.001). This model was validated using the K-fold cross-validation method; the accuracy was 0.815 ± 0.013 in training sets and 0.784 ± 0.019 in validation sets, indicating a good consistency for predicting the new-onset renal dysfunction risk. Finally, a nomogram based on this model was constructed to provide a quantitative tool to assess the individualized risk of short-term new-onset renal dysfunction. Conclusion: The model incorporating these markers and clinical features may have a high potential to predict the short-term risk of new-onset renal dysfunction.

Prevalence, Genetics and Evolutionary Properties of Eurasian Avian-like H1N1 Swine Influenza Viruses in Liaoning.

Swine influenza virus (SIV) is an important zoonosis pathogen. The 2009 pandemic of H1N1 influenza A virus (2009/H1N1) highlighted the importance of the role of pigs as intermediate hosts. Liaoning province, located in northeastern China, has become one of the largest pig-farming areas since 2016. However, the epidemiology and evolutionary properties of SIVs in Liaoning are largely unknown. We performed systematic epidemiological and genetic dynamics surveillance of SIVs in Liaoning province during 2020. In total, 33,195 pig nasal swabs were collected, with an SIV detection rate of 2%. Our analysis revealed that multiple subtypes of SIVs are co-circulating in the pig population in Liaoning, including H1N1, H1N2 and H3N2 SIVs. Furthermore, 24 H1N1 SIVs were confirmed to belong to the EA H1N1 lineage and divided into two genotypes. The two genotypes were both triple reassortant, and the predominant one with polymerase, nucleoprotein (NP), and matrix protein (M) genes originating from 2009/H1N1; hemagglutinin (HA) and neuraminidase (NA) genes originating from EA H1N1; and the nonstructural protein (NS) gene originating from triple reassortant H1N2 (TR H1N2) was detected in Liaoning for the first time. According to our evolutionary analysis, the EA H1N1 virus in Liaoning will undergo further genome variation.

A nomogram for predicting the risk of new-onset albuminuria based on baseline urinary ACR, orosomucoid, and HbA1c in patients with type 2 diabetes.

OBJECTIVES: Numerous biomarkers have been shown to be associated with albuminuria. However, few of them are valuable separate predictors of albuminuria development. This study aimed to develop a model for predicting the short-term risk of new-onset albuminuria in normoalbuminuric patients with type 2 diabetes (T2D). METHODS: 213 patients with T2D who were normoalbuminuric at the baseline were enrolled in this study. Basal levels of clinical characteristics and renal biomarkers including urinary orosomucoid (alpha-1-acid-glycoprotein, UORM), neutrophil gelatinase-associated lipocalin, retinol-binding protein, alpha-1-microglobulin, transferrin, and albumin-to-creatinine ratio (ACR) were utilized to analyze the association with the short-term risk of new-onset albuminuria. RESULTS: 19.72% of normoalbuminuric subjects at baseline progressed to albuminuria over the 2-year follow-up period. Except for NGAL, the basal levels of the other five renal biomarkers were significantly associated with new-onset albuminuria risk in the univariate analysis. In the multivariate logistic regression analysis using Forward: LR method, a model incorporating UORM/Cr, ACR, and HbA1c was established. Comparatively, this model had a higher potential to predict new-onset albuminuria risk compared with the single use of renal markers. In the validation of this model performed by 5-fold cross-validation method, the accuracy of this model was 0.818 ± 0.008 in the training sets, 0.827 ± 0.062 in the test sets, indicating a good capability for assessing albuminuria risk. Finally, a nomogram based on this model was constructed to facilitate its use in clinical practice. CONCLUSION: The combined analysis of UORM/Cr, ACR and HbA1c may be of potential value for predicting the short-term risk of new-onset albuminuria in such patients.

The appearance of volatile aromas in Tieguanyin tea with different elevations.

Chinese people have consistentlypreferred high mountain tea because specific flavors are memorable for them, and also, people have traditionally considered this type of tea to be good for health. Tieguanyin is one of the famous traditional Chinese tea that has ever-changing aromas. To illustrate the various characteristics in volatile fragment compounds from Tieguanyin tea, fresh tea leaves collected from different elevations (450, 650, and 900 m) were detected using GC-MS by solid-liquid extraction. The results showed that volatile aromatic compounds, such as benzyl alcohol, phenyl ethanol, and acetophenone, were the most abundant in tea leaves located with high elevation. Meanwhile, 1-hexanol, 1-nananol, and nanoic acid, as a type of aliphatic aroma, were more prevalent in low-elevation tea orchards. Catechols and alkaloids are largely cumulated in low- and high-elevation tea leaves, respectively. Our findings also showed that elemene was widely consisted of high-elevation tea metabolites. It provided practicality for the preparation of tea manufacturing in major Tieguanyin tea-producing regions.

Analysis of Trends in Insulin Utilization and Spending Across Canada From 2010 to 2015.

OBJECTIVES: To describe insulin utilization and spending across Canada and investigate how interprovincial variations in long-acting insulin uptake impact provincial spending. METHODS: We conducted a cross-sectional time-series analysis of insulin products dispensed nationally from January 1, 2010, to December 31, 2015, using data from IQVIA (Durham, North Carolina, United States). Analysis was stratified according to insulin type, payer and province. We report annual numbers for national insulin dispensing and spending and provincial numbers for publicly funded long-acting insulin dispensing and spending rates that are standardized by public drug beneficiary enrolment and diabetes prevalence. We report the percent of change of an annual provincial cost-to-utilization index of total insulin spending to total insulin dispensing between 2010 and 2015. RESULTS: Between 2010 and 2015, total insulin utilization increased 21% (4.4 million to 5.3 million prescriptions), and total insulin costs increased 54% ($345 million to $530 million) nationally. The national dispensing rate of long-acting insulin (+96%) and rapid-acting insulin (+38%) increased, while the national dispensing rate for intermediate-acting (-23%), short-acting (-37%) and premixed (-28%) insulins declined. Large interprovincial variation was observed for the rate of long-acting insulin uptake (range, Alberta, +1,505%; British Columbia, +27%) and the rate of long-acting insulin spending (Alberta, +2,177%; British Columbia, +44%) between 2010 and 2015 after standardization. Provinces with higher rates of long-acting insulin uptake experienced faster increases in their cost-to-utilization index (Alberta, +78%; British Columbia, +24%). CONCLUSIONS: Overall, the rate of uptake of long-acting insulins has increased nationally. Uptake varies widely among provinces and is likely to be associated with differential cost increases across public payers in Canada.

Network meta-analysis: an introduction for pharmacists.

Network meta-analysis is a new tool used to summarize and compare studies for multiple interventions, irrespective of whether these interventions have been directly evaluated against each other. Network meta-analysis is quickly becoming the standard in conducting therapeutic reviews and clinical guideline development. However, little guidance is available to help pharmacists review network meta-analysis studies in their practice. Major institutions such as the Cochrane Collaboration, Agency for Healthcare Research and Quality, Canadian Agency for Drugs and Technologies in Health, and National Institute for Health and Care Excellence Decision Support Unit have endorsed utilizing network meta-analysis to establish therapeutic evidence and inform decision making. Our objective is to introduce this novel technique to pharmacy practitioners, and highlight key assumptions behind network meta-analysis studies.