Blood-based epigenome-wide analyses of 19 common disease states: A longitudinal, population-based linked cohort study of 18,413 Scottish individuals.

BACKGROUND: DNA methylation is a dynamic epigenetic mechanism that occurs at cytosine-phosphate-guanine dinucleotide (CpG) sites. Epigenome-wide association studies (EWAS) investigate the strength of association between methylation at individual CpG sites and health outcomes. Although blood methylation may act as a peripheral marker of common disease states, previous EWAS have typically focused only on individual conditions and have had limited power to discover disease-associated loci. This study examined the association of blood DNA methylation with the prevalence of 14 disease states and the incidence of 19 disease states in a single population of over 18,000 Scottish individuals. METHODS AND FINDINGS: DNA methylation was assayed at 752,722 CpG sites in whole-blood samples from 18,413 volunteers in the family-structured, population-based cohort study Generation Scotland (age range 18 to 99 years). EWAS tested for cross-sectional associations between baseline CpG methylation and 14 prevalent disease states, and for longitudinal associations between baseline CpG methylation and 19 incident disease states. Prevalent cases were self-reported on health questionnaires at the baseline. Incident cases were identified using linkage to Scottish primary (Read 2) and secondary (ICD-10) care records, and the censoring date was set to October 2020. The mean time-to-diagnosis ranged from 5.0 years (for chronic pain) to 11.7 years (for Coronavirus Disease 2019 (COVID-19) hospitalisation). The 19 disease states considered in this study were selected if they were present on the World Health Organisation's 10 leading causes of death and disease burden or included in baseline self-report questionnaires. EWAS models were adjusted for age at methylation typing, sex, estimated white blood cell composition, population structure, and 5 common lifestyle risk factors. A structured literature review was also conducted to identify existing EWAS for all 19 disease states tested. The MEDLINE, Embase, Web of Science, and preprint servers were searched to retrieve relevant articles indexed as of March 27, 2023. Fifty-four of approximately 2,000 indexed articles met our inclusion criteria: assayed blood-based DNA methylation, had >20 individuals in each comparison group, and examined one of the 19 conditions considered. First, we assessed whether the associations identified in our study were reported in previous studies. We identified 69 associations between CpGs and the prevalence of 4 conditions, of which 58 were newly described. The conditions were breast cancer, chronic kidney disease, ischemic heart disease, and type 2 diabetes mellitus. We also uncovered 64 CpGs that associated with the incidence of 2 disease states (COPD and type 2 diabetes), of which 56 were not reported in the surveyed literature. Second, we assessed replication across existing studies, which was defined as the reporting of at least 1 common site in >2 studies that examined the same condition. Only 6/19 disease states had evidence of such replication. The limitations of this study include the nonconsideration of medication data and a potential lack of generalizability to individuals that are not of Scottish and European ancestry. CONCLUSIONS: We discovered over 100 associations between blood methylation sites and common disease states, independently of major confounding risk factors, and a need for greater standardisation among EWAS on human disease.

Comparison of the intraarticular osteotomy and the "window" osteotomy in the treatment of tibial plateau fracture involving depressed posterolateral fragments.

OBJECTIVES: The methods of reduction of depressed posterolateral fragments in tibial plateau fracture through anterolateral approaches remain controversial. This paper aimed to compare the intraarticular osteotomy technique and the "window" osteotomy technique for the reduction of depressed posterolateral fragments through anterolateral approach. METHOD: From January 2015 to January 2022, we retrospectively reviewed the data on patients with tibial plateau fracture involving depressed posterolateral fragments treated with the intraarticular osteotomy or the "window" osteotomy. 40 patients underwent the intraarticular osteotomy were divided into group A, while 36 patients underwent the "window" osteotomy were divided into group B. The operative time, bone grafting volume, fracture healing time, complication, reduction quality and postoperative functional results were compared between the two groups. RESULTS: The average follow-up duration was 16.6 ± 3.7 months. The average bone grafting volume for all patients in group B was essential larger than group A (p = 0.001). Compared to group B, patients in groups A had significantly shorter fracture healing time (p = 0.011). The depth of depressed articular surface, PSA and the radiographic evaluation at 2 days and 6 months after surgery in group A were significantly lower than group B (p<0.05). Based on the HSS knee-rating score, no significant difference in function results was found between the two groups (p>0.05). No significant difference was found in operation time and blood loss between the two groups (p>0.05). CONCLUSION: The intraarticular osteotomy could obtain satisfactory clinical results in tibial plateau fracture involving posterolateral fragments.

Scopoletin Reactivates Latent HIV-1 by Inducing NF-κB Expression without Global T Cell Activation.

Reversing HIV-1 latency promotes the killing of infected cells and is essential for cure strategies. However, current latency-reversing agents (LRAs) are not entirely effective and safe in activating latent viruses in patients. In this study, we investigated whether Scopoletin (6-Methoxy-7-hydroxycoumarin), an important coumarin phytoalexin found in plants with multiple pharmacological activities, can reactivate HIV-1 latency and elucidated its underlying mechanism. Using the Jurkat T cell model of HIV-1 latency, we found that Scopoletin can reactivate latent HIV-1 replication with a similar potency to Prostratin and did so in a dose- and time-dependent manner. Moreover, we provide evidence indicating that Scopoletin-induced HIV-1 reactivation involves the nuclear factor kappa B (NF-κB) signaling pathway. Importantly, Scopoletin did not have a stimulatory effect on T lymphocyte receptors or HIV-1 receptors. In conclusion, our study suggests that Scopoletin has the potential to reactivate latent HIV-1 without causing global T-cell activation, making it a promising treatment option for anti-HIV-1 latency strategies.

Using receiver operating characteristic curves to evaluate the diagnostic value of the combination of multislice spiral CT and alpha-fetoprotein levels for small hepatocellular carcinoma in cirrhotic patients.

BACKGROUND: The various combination of multiphase enhancement multislice spiral CT (MSCT) makes the diagnosis of a small hepatocellular carcinoma (sHCC) on the background of liver cirrhosis possible. This study was to explore whether the combination of MSCT enhancement scan and alpha-fetoprotein (AFP) level could increase the diagnostic efficiency for sHCC. METHODS: This study included 35 sHCC patients and 52 cirrhotic patients without image evidence of HCC as a control group. The diagnoses were made by three radiologists employing a 5-point rating scale, with postoperative pathologic results as the gold standard. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic value of the three MSCT combination modes (arterial phase+portal-venous phase, arterial phase+delayed phase, arterial phase+portal-venous phase+delayed phase) and AFP levels for sHCC on the background of liver cirrhosis. RESULTS: The area under ROC curve (AUC), sensitivity, and specificity of the combination of arterial phase+portal-venous phase+delayed phase were 0.93, 93%, and 82%, respectively. The average AUC of the arterial phase+portal-venous phase+delayed phase combination was significantly greater than that of the arterial phase+portal-venous phase (AUC=0.84, P=0.01) and arterial phase+delayed phase (AUC=0.85, P=0.03). Arterial phase+portal-venous phase had a smaller AUC (0.84) than arterial phase+delayed phase (0.85), but the difference was insignificant (P=0.15). After combining MSCT enhancement scan with AFP, the AUC, sensitivity, and specificity were 0.95, 94%, and 83%, respectively, indicating a greatly increased diagnostic efficiency for sHCC. CONCLUSIONS: The combination of AFP and 3 phases MSCT enhancement scan could increase the diagnostic efficiency for sHCC on the background of liver cirrhosis. The application of ROC curve analysis has provided a new method and reference in HCC diagnosis.

Enhanced riboflavin production by recombinant Bacillus subtilis RF1 through the optimization of agitation speed.

Dissolved oxygen is one of the most important bioprocess parameters that could affect cell growth and product formation, and it is easy to control by changing agitation speed. In this work, the effects of agitation speed on the performance of riboflavin production by recombinant Bacillus subtilis RF1 was investigated in fed-batch fermentation. The lower agitation speed (600 rpm) was beneficial for cell growth and riboflavin biosynthesis in the initial phase of fermentation process. While, during the later phase, higher agitation speed (900 rpm) was favor for cell growth and riboflavin biosynthesis. Thus, a two-stage agitation speed control strategy was proposed based on kinetic analysis, in which the agitation speed was controlled at 600 rpm in the first 26 h and then switched to 900 rpm to maintain high µ for cell growth and high q(p) for riboflavin production during the entire fermentation process. However, it was observed that a sharp increase of agitation speed resulted in an adverse effect on cell growth and riboflavin synthesis within a short time. To avoid this phenomenon, a multi-stage agitation speed control strategy was set up based on the two-stage control strategy, the maximum concentration of riboflavin reached 9.4 g l(-1) in 48 h with the yield of 0.051 g g(-1) by applying this strategy, which were 20.5 and 21.4% over the best results controlled by constant agitation speeds.

Effective in vivo reactivation of HIV-1 latency reservoir via oral administration of EK-16A-SNEDDS.

The latent reservoir of human immunodeficiency virus (HIV) is a major obstacle in the treatment of acquired immune deficiency syndrome (AIDS). The "shock and kill" strategy has emerged as a promising approach for clearing HIV latent reservoirs. However, current latency-reversing agents (LRAs) have limitations in effectively and safely activating the latent virus and reducing the HIV latent reservoirs in clinical practice. Previously, EK-16A was extracted from Euphorbia kansui, which had the effect of interfering with the HIV-1 latent reservoir and inhibiting HIV-1 entry. Nevertheless, there is no suitable and efficient EK-16A oral formulation for in vivo delivery and clinical use. In this study, an oral EK-16A self-nanoemulsifying drug delivery system (EK-16A-SNEDDS) was proposed to "shock" the HIV-1 latent reservoir. This system aims to enhance the bioavailability and delivery of EK-16A to various organs. The composition of EK-16A-SNEDDS was optimized through self-emulsifying grading and ternary phase diagram tests. Cell models, pharmacokinetic experiments, and pharmacodynamics in HIV-1 latent cell transplant animal models suggested that EK-16A-SNEDDS could be absorbed by the gastrointestinal tract and enter the blood circulation after oral administration, thereby reaching various organs to activate latent HIV-1. The prepared EK-16A-SNEDDS demonstrated safety and efficacy, exhibited high clinical experimental potential, and may be a promising oral preparation for eliminating HIV-1 latent reservoirs.

Epigenetic Contributions to Clinical Risk Prediction of Cardiovascular Disease.

BACKGROUND: Cardiovascular disease (CVD) is among the leading causes of death worldwide. The discovery of new omics biomarkers could help to improve risk stratification algorithms and expand our understanding of molecular pathways contributing to the disease. Here, ASSIGN-a cardiovascular risk prediction tool recommended for use in Scotland-was examined in tandem with epigenetic and proteomic features in risk prediction models in ≥12 657 participants from the Generation Scotland cohort. METHODS: Previously generated DNA methylation-derived epigenetic scores (EpiScores) for 109 protein levels were considered, in addition to both measured levels and an EpiScore for cTnI (cardiac troponin I). The associations between individual protein EpiScores and the CVD risk were examined using Cox regression (ncases≥1274; ncontrols≥11 383) and visualized in a tailored R application. Splitting the cohort into independent training (n=6880) and test (n=3659) subsets, a composite CVD EpiScore was then developed. RESULTS: Sixty-five protein EpiScores were associated with incident CVD independently of ASSIGN and the measured concentration of cTnI (P<0.05), over a follow-up of up to 16 years of electronic health record linkage. The most significant EpiScores were for proteins involved in metabolic, immune response, and tissue development/regeneration pathways. A composite CVD EpiScore (based on 45 protein EpiScores) was a significant predictor of CVD risk independent of ASSIGN and the concentration of cTnI (hazard ratio, 1.32; P=3.7×10-3; 0.3% increase in C-statistic). CONCLUSIONS: EpiScores for circulating protein levels are associated with CVD risk independent of traditional risk factors and may increase our understanding of the etiology of the disease.

A unified framework for estimating country-specific cumulative incidence for 18 diseases stratified by polygenic risk.

Polygenic scores (PGSs) offer the ability to predict genetic risk for complex diseases across the life course; a key benefit over short-term prediction models. To produce risk estimates relevant to clinical and public health decision-making, it is important to account for varying effects due to age and sex. Here, we develop a novel framework to estimate country-, age-, and sex-specific estimates of cumulative incidence stratified by PGS for 18 high-burden diseases. We integrate PGS associations from seven studies in four countries (N = 1,197,129) with disease incidences from the Global Burden of Disease. PGS has a significant sex-specific effect for asthma, hip osteoarthritis, gout, coronary heart disease and type 2 diabetes (T2D), with all but T2D exhibiting a larger effect in men. PGS has a larger effect in younger individuals for 13 diseases, with effects decreasing linearly with age. We show for breast cancer that, relative to individuals in the bottom 20% of polygenic risk, the top 5% attain an absolute risk for screening eligibility 16.3 years earlier. Our framework increases the generalizability of results from biobank studies and the accuracy of absolute risk estimates by appropriately accounting for age- and sex-specific PGS effects. Our results highlight the potential of PGS as a screening tool which may assist in the early prevention of common diseases.

Coumarin 6 staining method to detect microplastics.

Microplastics have contaminated the ocean in large quantities and are widely distributed throughout the world. Thus, our understanding of the concentration of microplastics in various environments should be increased. However, current methods to detect microplastics require considerable effort and expensive equipment. In this study, we developed a fluorescence staining technique using coumarin 6 and examined its effectiveness. A mixture of acetone and ethanol was used as the solvent, and 10 different types of plastics were able to be stained with coumarin 6. The fluorescence peak for coumarin 6 staining was approximately 500 nm for each plastic type. The optimal immersion time and coumarin 6 concentration for staining were determined to be 60 min and 1 mg L-1, respectively. Using this technique, we were able to stain all of the microplastics obtained from samples collected in Tokyo Bay seawater.

[Osteotomy of non-core weight-bearing area of the lateral tibial plateau, reduction, and internal fixation in treatment of tibial plateau fractures involving posterolateral column collapse].

Objective: To investigate the effectiveness of osteotomy of non-core weight-bearing area of the lateral tibial plateau, reduction, and internal fixation in the treatment of tibial plateau fractures involving posterolateral column collapse. Methods: A clinical data of 23 patients with tibial plateau fractures involving posterolateral column collapse, who had undergone osteotomy of non-core weight-bearing area of the lateral tibial plateau, reduction, and internal fixation between January 2015 and June 2021, was retrospectively analyzed. There were 14 males and 9 females with an average age of 42.6 years ranging from 26 to 62 years. The causes of injury included traffic accident in 16 cases, falling from height in 5 cases, and other injuries in 2 cases. According to Schatzker classification, there were 15 cases of type Ⅴ and 8 cases of type Ⅵ. The time from injury to operation was 4-8 days with an average of 5.9 days. The operation time, intraoperative blood loss, fracture healing time, and complications were recorded. The depth of articular surface collapse of posterolateral column and posterior inclination angle (PSA) of the tibial plateau were compared before operation and at 2 days and 6 months after operation; fracture reduction of tibial plateau fracture was evaluated by Rasmussen anatomic score. The recovery of knee function was evaluated by Hospital for Special Surgery (HSS) score at 2 days and 6 months after operation. Results: All 23 patients were completed the operation successfully. The operation time was 120-195 minutes, with an average of 152.8 minutes; the intraoperative blood loss was 50-175 mL, with an average of 109.5 mL. All patients were followed up 12-24 months, with an average of 16.7 months. One patient had superficial wound infection after operation, and the incision healed after dressing change; primary healing of incision of other patients was obtained. The fracture healing time was 12-18 weeks, with an average of 13.7 weeks. No failure of internal fixation, varus and valgus deformity of the knee joint, and instability of the knee joint was found at last follow-up. One patient developed joint stiffness and the range of motion of the knee joint was 10°-100°; the range of motion of the knee joint of other patients was 0°-125°. At 2 days and 6 months after operation, the depth of articular surface collapse of posterolateral column, PSA, and Rasmussen anatomic scores significantly improved when compared with those before operation ( P<0.05). There was no significant difference between the two postoperative time points ( P>0.05). The HSS score at 6 months after operation was significantly higher than that at 2 days after operation ( P<0.05). Conclusion: For tibial plateau fractures involving posterolateral column collapse, reduction and internal fixation through osteotomy of non-core weight-bearing area of the lateral tibial plateau has the advantages of fully expose the posterolateral column fragment, good articular surface reduction, sufficient bone grafting, and fewer postoperative complications. It is beneficial to restore knee joint function and can be widely used in clinic.

Three-Dimensional Printed Plate Template Versus Hemipelvis Model in Patient-Specific Plate Preparation for Posterior Wall Acetabular Fractures.

OBJECTIVE: The application of three-dimensional (3D) printing technology in the management of posterior wall acetabular fractures can greatly reduce surgical invasiveness and operative time and simplify the procedure of reconstruction plate contouring, but the cost and time of patient-specific plate preparation on the basis of traditional 3D-printed pelvis model should not be neglected. We described a new method for patient-specific plate preparation by using 3D-printed plate template. The study aimed to assess the effectiveness and feasibility of the 3D-printed plate template in patient-specific plate preparation for posterior wall acetabular fractures. METHODS: A total of 65 cases of posterior wall acetabular fractures with surgical treatment from December 2012 to December 2020 were chosen. According to the different plate contouring methods, the 65 cases were divided into three groups, which were group A (21 cases), group B (20 cases), and group C (24 cases). In group A, the 3D-printed plate template was used to contour the patient-specific reconstruction plate before surgery, whereas the 3D-printed hemipelvis model was adopted for group B. In group C, the reconstruction plate was contoured intraoperatively. Among the three groups, the instrumentation time, surgical time, blood loss, patient-specific plate preparation time, complications, reduction quality, and hip function were compared. The Kruskal-Wallis test was used to analyze the reduction quality and hip function among three groups. RESULTS: In comparison with group C, patients in groups A and B were featured by obviously shorter instrumentation time (-22, -23 min), shorter surgical time (-46, -44 min), and less intraoperative blood loss (-110, -122 mL). Compared to the hemipelvis model in group B (2.29 ± 0.56 vs. 12.70 ± 3.79 days), the 3D printing time for plate templates in group A was significantly shorter. The reduction quality and hip function had no obvious statistical difference among the three groups. The complication rate within group A (3/21) and group B (3/20) were both slightly lower than group C (5/24), with no obvious difference. CONCLUSIONS: Both the patient-specific pre-contoured plate fixation methods based on the 3D-printed hemipelvis model and plate template can achieve satisfactory clinical efficacy, with the advantage of shorter instrumentation and surgical time, and less intraoperative blood loss. However, 3D printing of plate template is easier and less time-consuming, considering the shorter time and less cost for 3D printing of physical model.

Development and validation of DNA methylation scores in two European cohorts augment 10-year risk prediction of type 2 diabetes.

Type 2 diabetes mellitus (T2D) presents a major health and economic burden that could be alleviated with improved early prediction and intervention. While standard risk factors have shown good predictive performance, we show that the use of blood-based DNA methylation information leads to a significant improvement in the prediction of 10-year T2D incidence risk. Previous studies have been largely constrained by linear assumptions, the use of cytosine-guanine pairs one-at-a-time and binary outcomes. We present a flexible approach (via an R package, MethylPipeR) based on a range of linear and tree-ensemble models that incorporate time-to-event data for prediction. Using the Generation Scotland cohort (training set ncases = 374, ncontrols = 9,461; test set ncases = 252, ncontrols = 4,526) our best-performing model (area under the receiver operating characteristic curve (AUC) = 0.872, area under the precision-recall curve (PRAUC) = 0.302) showed notable improvement in 10-year onset prediction beyond standard risk factors (AUC = 0.839, precision-recall AUC = 0.227). Replication was observed in the German-based KORA study (n = 1,451, ncases = 142, P = 1.6 × 10-5).

Traditional Mongolian medicine (HHQG) attenuates CCl4-induced acute liver injury through inhibiting monocyte/macrophage infiltration via the p-P38/p-JNK pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Honghua Qinggan 13 Flavor Pills (HHQG), whose Mongolian name is Guri Gumu-13, is a traditional Mongolian medicine, that was stated in the "Diagnosis and Treatment of Ming Medical Code". The HHQG has been included in the Mongolian Medicine Division of the Ministry of Health Drug Standards (1998 edition). Based on our clinical expertise, HHQG demonstrated satisfactory therapeutic effects in hepatitis and liver failure. However, the pharmacological effects and potential mechanisms of HHQG have not been investigated. AIM OF THE STUDY: In this study, we combined network pharmacology, transcriptomics, and molecular biology to detect the underlying mechanism for the effect of HHQG on acute liver injury in mice. MATERIALS AND METHODS: Network pharmacology was used to explore the pathways involved in the protective effect HHQG in acute liver injury. This effect was further verified by injecting carbon tetrachloride (CCl4; 10 mL/kg, i.p.) to induce acute liver injury in mice. Serum markers of liver injury, morphology, histology, and monocyte/macrophage infiltration in the liver tissue were investigated. Transcriptomics further defined the HHQG targets. Transwell analysis was performed to confirm that HHQG inhibited monocyte/macrophage RAW.264.7 infiltration. qPCR and Western blot were performed to explore the mechanism of action of HHQG. RESULTS: Network pharmacology showed that HHQG exerted anti-oxidative and anti-inflammatory effects and promoted metabolic effects against acute liver injury. Pretreatment of mice with HHQG significantly maintained their body weight and decreased serum tumor necrosis factor-alpha (TNF-α) levels induced by CCl4 treatment in vivo. Histopathological examination further confirmed that HHQG protected the liver cells from CCl4-induced damage. Importantly, HHQG significantly inhibited CCl4-induced monocyte/macrophage infiltration. Transcriptomic analysis revealed that HHQG significantly reduced the expression of chemokines and cell adhesion molecules. We determined that HHQG significantly downregulated the expression of the key chemokine (monocyte chemokine protein-1, CCL2) at the gene and protein levels. Further research showed that HHQG inhibited chemokine production in hepatocytes by inhibiting the p-P38 and p-JNK pathways, thereby reducing monocyte/macrophage infiltration. CONCLUSIONS: These combined data showed that HHQG alleviated acute liver injury in mice, and further verified that HHQG exerted protective effects by inhibiting the production of CCL2 and reducing the infiltration of monocyte/macrophage by inhibiting the p-P38 and p-JNK pathways.

Design and Synthesis of Dipeptidomimetic Isocyanonaphthalene as Enhanced-Fluorescent Chemodosimeter for Sensing Mercury Ion and Living Cells.

A novel valine-based isocyanonaphthalene (NpI) was designed and synthesized by using an easy method and enabled the selective fluorescence detection of Hg2+. The chemodosimeter can display an immediate turn-on fluorescence response (500-fold) towards target metal ions upon the Hg2+-mediated conversion of isocyano to amino within NpI. Based on this specific reaction, the fluorescence-enhancement probe revealed a high sensitivity toward Hg2+ over other common metal ions and exhibited excellent aqueous solubility, good antijamming capability, high sensitivity (detection limit: 14.2 nM), and real-time detection. The response mechanism of NpI was supported by NMR spectroscopy, MS analysis and DFT theoretical calculation using various techniques. Moreover, a dipeptidomimetic NpI probe was successfully applied to visualize intracellular Hg2+ in living cells and monitor Hg2+ in real water samples with good recoveries and small relative standard deviations.

Upregulated miR-322-5p regulates cell cycle and promotes cell proliferation and apoptosis by directly targeting Wee1 in mice liver injury.

Liver injury from any number of causes (e.g. chemical material, drugs and diet, viral infection) is a global health problem, and its mechanism is not clearly understood. MicroRNAs (miRNAs) expression profiling is gaining popularity because miRNAs, as key regulators in gene expression networks, can influence many biological processes and have also shown promise as biomarkers for disease. Previous studies reported the regulation effects of miRNAs in liver injury, whereas function and molecular mechanisms of miR-322-5p were still unclear. Therefore, our study focused on the biological role of miR-322-5p in carbon tetrachloride (CCl4)-induced liver injury proliferation, apoptosis, and cell cycle. A mouse model of CCl4-induced liver injury was established, and the transcriptomes and miRNAs transcriptomes of 2d and 5d liver tissues after injury were sequenced. The expression of miR-322-5p and the cell cycle genes were detected in liver tissues and Hepa1-6 cell line by miRNA RT-PCR, qRT-PCR. The effects of miR-322-5p on liver cell proliferation, cell cycle and apoptosis were evaluated using MTS assays and flow cytometry analysis. The relationship between miR-322-5p and Wee1 was predicted and confirmed by bioinformatics analysis and a dual luciferase reporter assay. Functional experiments, including an MTS assay and flow cytometric analysis, were performed to study the effects of Wee1. MiR-322-5p was upregulated in injury liver tissues, and downregulated miR-322-5p was proved to inhibit proliferation, apoptosis and arrest cell cycle at G2/M in vitro. The dual-luciferase reporter assay results indicated that miR-322-5p has a binding site at position 285 in the Wee1 3´UTR. The effects of miR-322-5p in proliferation and cell cycle regulation can be abolished by Wee1 through rescue experiments. By directly targeting Wee1 influenced the expression of several cell cycle factors, including Cyclin dependent kinase 1 (Cdk1), cyclin B1 (Ccnb1) and Cell division cyclin 25C (Cdc25C). MiR-322-5p may function as a suppressive factor by negatively controlling Wee1, thus, highlighting the potential role of miR-322-5p as a therapeutic target for liver injury.Abbreviations: ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; GSH: Glutathione, γ-glutamyl cysteinel + glycine; CCl4: Carbon tetrachloride; HE: Haematoxylin and eosin; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Epigenetic scores for the circulating proteome as tools for disease prediction.

Protein biomarkers have been identified across many age-related morbidities. However, characterising epigenetic influences could further inform disease predictions. Here, we leverage epigenome-wide data to study links between the DNA methylation (DNAm) signatures of the circulating proteome and incident diseases. Using data from four cohorts, we trained and tested epigenetic scores (EpiScores) for 953 plasma proteins, identifying 109 scores that explained between 1% and 58% of the variance in protein levels after adjusting for known protein quantitative trait loci (pQTL) genetic effects. By projecting these EpiScores into an independent sample (Generation Scotland; n = 9537) and relating them to incident morbidities over a follow-up of 14 years, we uncovered 137 EpiScore-disease associations. These associations were largely independent of immune cell proportions, common lifestyle and health factors, and biological aging. Notably, we found that our diabetes-associated EpiScores highlighted previous top biomarker associations from proteome-wide assessments of diabetes. These EpiScores for protein levels can therefore be a valuable resource for disease prediction and risk stratification.

Although our genetic code does not change throughout our lives, our genes can be turned on and off as a result of epigenetics. Epigenetics can track how the environment and even certain behaviors add or remove small chemical markers to the DNA that makes up the genome. The type and location of these markers may affect whether genes are active or silent, this is, whether the protein coded for by that gene is being produced or not. One common epigenetic marker is known as DNA methylation. DNA methylation has been linked to the levels of a range of proteins in our cells and the risk people have of developing chronic diseases. Blood samples can be used to determine the epigenetic markers a person has on their genome and to study the abundance of many proteins. Gadd, Hillary, McCartney, Zaghlool et al. studied the relationships between DNA methylation and the abundance of 953 different proteins in blood samples from individuals in the German KORA cohort and the Scottish Lothian Birth Cohort 1936. They then used machine learning to analyze the relationship between epigenetic markers found in people's blood and the abundance of proteins, obtaining epigenetic scores or 'EpiScores' for each protein. They found 109 proteins for which DNA methylation patterns explained between at least 1% and up to 58% of the variation in protein levels. Integrating the 'EpiScores' with 14 years of medical records for more than 9000 individuals from the Generation Scotland study revealed 130 connections between EpiScores for proteins and a future diagnosis of common adverse health outcomes. These included diabetes, stroke, depression, various cancers, and inflammatory conditions such as rheumatoid arthritis and inflammatory bowel disease. Age-related chronic diseases are a growing issue worldwide and place pressure on healthcare systems. They also severely reduce quality of life for individuals over many years. This work shows how epigenetic scores based on protein levels in the blood could predict a person's risk of several of these diseases. In the case of type 2 diabetes, the EpiScore results replicated previous research linking protein levels in the blood to future diagnosis of diabetes. Protein EpiScores could therefore allow researchers to identify people with the highest risk of disease, making it possible to intervene early and prevent these people from developing chronic conditions as they age.

Application of Field-of-View Optimized and Constrained Undistorted Single Shot (FOCUS) with Intravoxel Incoherent Motion (IVIM) in 3T in Locally Advanced Rectal Cancer.

PURPOSE: To evaluate the efficacy of field-of-view (FOV) optimized and constrained undistorted single shot (FOCUS) with IVIM in 3T MRI in the grading of patients with locally advanced rectal cancer. METHODS: From January 1st to December 31st, 2019, patients with locally advanced rectal cancer were retrieved. FOCUS DWI and FOCUS IVIM were obtained. Apparent diffusion coefficient (ADC) and IVIM parameters including mean true diffusion coefficient (D), pseudodiffusion coefficient associated with blood flow (D∗), and perfusion fraction (f) of the tumor parenchyma and normal rectal wall, as well as the normalized tumor parameters by corresponding normal intestinal wall parameters (ADCNOR, D NOR, D∗NOR, and f NOR), were compared between the well/moderately differentiated and poorly differentiated groups by Student's t-test. The relationship between the above parameters and the histologic grade was analyzed using Spearman's correlation test, with the ROC curve generated. RESULTS: Eighty-eight patients (aged 31 to 77 years old, mean = 56) were included for analysis. D tumor and f tumor were positively correlated with the tumor grade (r = 0.483, p < 0.001 and r = 0.610, p < 0.001, respectively). All the normalized parameters (ADCNOR, D NOR, D∗NOR, and f NOR) were positively correlated with the tumor grade (r = 0.267, p = 0.007; r = 0.564, p = 0.001; r = 0.414, p = 0.005; and r = 0.605, p < 0.001, respectively). The best discriminative parameter was the f tumor value, and the area under the ROC curve was 0.927. With a cut-off value of 22.0%, f tumor had a sensitivity of 88.9% and a specificity of 100%. CONCLUSION: FOCUS IVIM-derived parameters and normalized parameters are useful for predicting the histologic grade in rectal cancer patients.

Associations between Dietary Nutrient Intakes and Hepatic Lipid Contents in NAFLD Patients Quantified by ¹H-MRS and Dual-Echo MRI.

Dietary habits are crucial in the progression of hepatic lipid accumulation and nonalcoholic fatty liver disease (NAFLD). However, there are limited studies using ¹H-magnetic resonance spectroscopy (¹H-MRS) and dual-echo in-phase and out-phase magnetic resonance spectroscopy imaging (dual-echo MRI) to assess the effects of dietary nutrient intakes on hepatic lipid contents. In the present study, we recruited 36 female adults (NAFLD:control = 19:17) to receive questionnaires and medical examinations, including dietary intakes, anthropometric and biochemical measurements, and ¹H-MRS and dual-echo MRI examinations. NAFLD patients were found to consume diets higher in energy, protein, fat, saturated fatty acid (SFA), and polyunsaturated fatty acid (PUFA). Total energy intake was positively associated with hepatic fat fraction (HFF) and intrahepatic lipid (IHL) after adjustment for age and body-mass index (BMI) (HFF: ß = 0.24, p = 0.02; IHL: ß = 0.38, p = 0.02). Total fat intake was positively associated with HFF and IHL after adjustment for age, BMI and total energy intake (HFF: ß = 0.36, p = 0.03; IHL: ß = 0.42, p = 0.01). SFA intake was positively associated with HFF and IHL after adjustments (HFF: ß = 0.45, p = 0.003; IHL: ß = 1.16, p = 0.03). In conclusion, hepatic fat content was associated with high energy, high fat and high SFA intakes, quantified by ¹H-MRS and dual-echo MRI in our population. Our findings are useful to provide dietary targets to prevent the hepatic lipid accumulation and NAFLD.