Identifying the importance of PCK1 in maintaining ileal epithelial barrier integrity in Crohn's disease.

BACKGROUND: Crohn's disease (CD) is marked by disruption of intestinal epithelial barrier, with unclear underlying molecular mechanisms. This study aimed to investigate key genes regulating the intestinal barrier in CD patients. METHODS: Differential gene expression analysis and gene set enrichment analysis were conducted to identify potential key genes involved in CD within the GEO database. Single-cell RNA sequencing from ileum samples in GSE134809 of 59,831 inflamed and uninflamed cells from 11 CD patients and microarray data from ileal tissues in GSE69762 (3 controls and 4 CD patients) and GSE75214 (11 controls and 51 CD patients) with GSE179285 (49 uninflamed and 33 inflamed from CD patients) as the validation set. Protein-protein interaction and logistic regression analyses identified key downregulated genes in CD. A key gene was then investigated through immunohistochemistry of ileal tissues from 5 CD patients and in the Caco-2 cell line with RNA interference and treatment with IFN-γ and TNF-α to stimulate inflammation. RESULTS: Single-cell RNA-seq identified 33 genes and microarray identified 167 genes with significant downregulation in inflamed CD samples. PCK1 was identified and validated as one of the most promising candidate genes. Reduced PCK1 expression was evident in inflamed ileal tissues. In vitro, knockdown of PCK1 resulted in decreased cell viability, increased apoptosis, and reduced nectin-2 production, while combination of IFN-γ and TNF-α significantly reduced PCK1. CONCLUSIONS: PCK1 is downregulated in inflamed ileal tissues of CD patients and may be a key factor in maintaining epithelial integrity during inflammation in Crohn's disease.

Amylose content controls the V-type structural formation and in vitro digestibility of maize starch-resveratrol complexes and their effect on human gut microbiota.

The chain structure of starch affects its interaction with polyphenol molecules which in turn determines the nutritional function of starch. In this study, starch with different amylose content including waxy maize starch (WMS), normal maize starch (NMS) and G50 high-amylose maize starch (G50) were selected to complex with resveratrol (RA) in high-pressure homogenization (HPH) environment, and structural changes of the complexes, together with their effects on in vitro digestibility and gut microbiota were discussed. The results showed that with increasing amylose content, RA could form more inclusion complex with starch through non-covalent bonds accompanied by the increased single helix structure, V-type crystalline structure, compact nano-aggregates and total ordered structure content, which thus endowed the complex lower digestibility and intestinal probiotic function. Notably, when RA addition reached 3 %, the resistant starch (RS) content of HP-G50-3 % rose to 29.2 %, correspondingly increased the relative abundance of beneficial gut microbiota such as Megamonas and Bifidobacterium, as well as the total short-chain fatty acids (SCFAs) content. Correlation analysis showed that V-type crystalline structure positively correlated with the growth of Pediococcu and Blautia (p < 0.05) for producing SCFAs. These findings provided feasible ideas for the development of personalized nutritional starch-based foods.

Heat-moisture treatment enhances the ordered degree of starch structure in whole chestnut flour and alters its gut microbiota modulation in mice fed with high-fat diet.

Currently, chestnuts attract more attention among consumers due to its rich nutritional functions, but systematic evaluation on the effect of thermal processing on its nutritional value is still limited. In this work, based on results of microstructural properties that heat-moisture treatment (HMT) could enhance the total ordered degree of starch structure in whole chestnut flour (CN) and promote the formation of anti-enzymatic component, in vitro experiment was then conducted and confirmed that HMT could significantly reduce the predicted glycemic index (pGI) of CN from 75.6 to 64.3 and improve its dietary fiber content from 7.06 to 13.42 g/100 g (p < 0.05). Further dietary intervention studies with CN and heat-moisture treated CN (HMT-CN) supplementation on the high-fat diet (HFD) consuming mice were discussed in terms of gut microbiota and its metabolites changes. The results showed that both CN and HMT-CN significantly resisted the weight gain induced by HFD, while HMT-CN had better serum lipid regulation effect. However, they had different effects on the gut metabolism pathways, among which CN inhibited the production of stearamine by promoting the proliferation of Dubosiella, while HMT-CN contributed to the growth of Lachnoclostridium, Desulfovibrio, and Faecalibaculum which stimulated the formation of associated metabolites including jwh-018-d11, valylproline, tetranor-12(S)-HETE, and PA (3:0/18:0). Overall, these discoveries could provide basic data for the effective utilization of CN in food industry processing.

Serum ferritin and the risk of short-term mortality in critically ill patients with chronic heart failure: a retrospective cohort study.

Background: Serum ferritin levels are associated with a higher risk of incident heart failure (HF). Whether serum ferritin levels, either increased or decreased, predict the risk of mortality in individuals with chronic heart failure (CHF) remains unknown. Objectives: This study aimed to clarify the potential predictive significance of serum ferritin levels in assessing the short-term mortality in critically ill patients with chronic heart failure (CHF). Methods: Critically ill patients with CHF were identified from the Multiparameter Intelligent Monitoring in Intensive Care III and IV (MIMIC III and IV) databases. Linear and logistic regression models and Cox proportional hazards models were applied to assess the associations between serum ferritin and survival. Results: A total of 1,739 and 2,322 patients with CHF identified from the MIMIC III and IV databases, respectively, fulfilled the inclusion criteria. In the MIMIC III group, compared with the reference group (serum ferritin ≥70 and <500 ng/mL), serum ferritin ≥1000 ng/mL was a significant predictor of 28-day (odds ratio [OR], 1.76; 95% confidence interval [CI], 1.14-2.72) and 90-day mortality (OR, 1.64; 95% CI, 1.13-2.39). The results from the Cox regression and Kaplan-Meier curves revealed similar results. In the MIMIC IV group, serum ferritin ≥1000 ng/mL was a significant predictor of in-hospital (OR, 1.70; 95% CI, 1.18-2.46), 28-day (OR, 1.83; 95% CI, 1.24-2.69), and 90-day mortality (OR, 1.57; 95% CI, 1.11-2.22) after adjusting for confounding factors. Conclusion: High ferritin levels (≥1000 ng/mL) were associated with increased short-term mortality in critically ill patients with CHF, indicating that serum ferritin may serve as a useful prognostic marker for CHF.

The emerging roles of HDACs and their therapeutic implications in cancer.

Deregulation of protein post-translational modifications is intensively involved in the etiology of diseases, including degenerative diseases, inflammatory injuries, and cancers. Acetylation is one of the most common post-translational modifications of proteins, and the acetylation levels are controlled by two mutually antagonistic enzyme families, histone acetyl transferases (HATs) and histone deacetylases (HDACs). HATs loosen the chromatin structure by neutralizing the positive charge of lysine residues of histones; whereas HDACs deacetylate certain histones, thus inhibiting gene transcription. Compared with HATs, HDACs have been more intensively studied, particularly regarding their clinical significance. HDACs extensively participate in the regulation of proliferation, migration, angiogenesis, immune escape, and therapeutic resistance of cancer cells, thus emerging as critical targets for clinical cancer therapy. Compared to HATs, inhibitors of HDAC have been clinically used for cancer treatment. Here, we enumerate and integratethe mechanisms of HDAC family members in tumorigenesis and cancer progression, and address the new and exciting therapeutic implications of single or combined HDAC inhibitor (HDACi) treatment.

Tricine as a Novel Cryoprotectant with Osmotic Regulation, Ice Recrystallization Inhibition and Antioxidant Properties for Cryopreservation of Red Blood Cells.

The cryopreservation of red blood cells (RBCs) plays a key role in blood transfusion therapy. Traditional cryoprotectants (CPAs) are mostly organic solvents and may cause side effects to RBCs, such as hemolysis and membrane damage. Therefore, it is necessary to find CPAs with a better performance and lower toxicity. Herein, we report for the first time that N-[Tri(hydroxymethyl)methyl]glycine (tricine) showed a great potential in the cryopreservation of sheep RBCs. The addition of tricine significantly increased the thawed RBCs' recovery from 19.5 ± 1.8% to 81.2 ± 8.5%. The properties of thawed RBCs were also maintained normally. Through mathematical modeling analysis, tricine showed a great efficiency in cryopreservation. We found that tricine had a good osmotic regulation capacity, which could mitigate the dehydration of RBCs during cryopreservation. In addition, tricine inhibited ice recrystallization, thereby decreasing the mechanical damage from ice. Tricine could also reduce oxidative damage during freezing and thawing by scavenging reactive oxygen species (ROS) and maintaining the activities of endogenous antioxidant enzymes. This work is expected to open up a new path for the study of novel CPAs and promote the development of cryopreservation of RBCs.

Environmental enteric dysfunction induces regulatory T cells that inhibit local CD4+ T cell responses and impair oral vaccine efficacy.

Environmental enteric dysfunction (EED) is a gastrointestinal inflammatory disease caused by malnutrition and chronic infection. EED is associated with stunting in children and reduced efficacy of oral vaccines. To study the mechanisms of oral vaccine failure during EED, we developed a microbiota- and diet-dependent mouse EED model. Analysis of E. coli-labile toxin vaccine-specific CD4+ T cells in these mice revealed impaired CD4+ T cell responses in the small intestine and but not the lymph nodes. EED mice exhibited increased frequencies of small intestine-resident RORγT+FOXP3+ regulatory T (Treg) cells. Targeted deletion of RORγT from Treg cells restored small intestinal vaccine-specific CD4 T cell responses and vaccine-mediated protection upon challenge. However, ablation of RORγT+FOXP3+ Treg cells made mice more susceptible to EED-induced stunting. Our findings provide insight into the poor efficacy of oral vaccines in EED and highlight how RORγT+FOXP3+ Treg cells can regulate intestinal immunity while leaving systemic responses intact.

Venous thromboembolic events in patients with COVID-19: a systematic review and meta-analysis.

BACKGROUND: High incidence of venous thromboembolic complications in coronavirus disease 2019 (COVID-19) patients was noted recently. OBJECTIVE: This study aimed to explore the factors associated with prevalence of venous thromboembolism (VTE) in COVID-19 patients. METHODS: A literature search was conducted in several online databases. Fixed effects meta-analysis was performed for the factors associated with prevalence of VTE in COVID-19 patients. RESULTS: A total of 39 studies were analysed in this analysis. The incidence of pulmonary embolism and VTE in severe COVID-19 patients were 17% (95% CI, 13-21%) and 42% (95% CI, 25-60%), respectively. VTE were more common among individuals with COVID-19 of advance age. Male COVID-19 patients are more likely to experience VTE. Higher levels of white blood cell (WBC; WMD = 1.34 × 109/L; 95% CI, 0.84-1.84 × 109/L), D-dimer (WMD = 4.21 µg/ml; 95% CI, 3.77-4.66 µg/ml), activated partial thromboplastin time (APTT; WMD = 2.03 s; 95% CI, 0.83-3.24 s), fibrinogen (WMD = 0.49 µg/ml; 95% CI, 0.18-0.79 g/L) and C-reactive protein (CRP; WMD = 21.89 mg/L; 95% CI, 11.44-32.34 mg/L) were commonly noted in COVID-19 patients with VTE. Patients with lower level of lymphocyte (WMD = -0.15 × 109/L; 95% CI, -0.23--0.07 × 109/L) was at high risk of developing VTE. The incidence of severe condition (OR = 2.66; 95% CI, 1.95-3.62) was more likely to occur among COVID-19 patients who developed VTE. CONCLUSION: VTE is a common complication in severe COVID-19 patients and thromboembolic events are also associated with adverse outcomes.

Intake of Anthocyanins and Gastric Cancer Risk: A Comprehensive Meta-Analysis on Cohort and Case-Control Studies.

This meta-analysis aimed to explore the association between anthocyanins intake and the risk of gastric cancer. All the relative articles have been searched in the online databases, including PubMed, EMBASE, Web of Science, and the Cochrane Library until June 11th, 2018. Risk ratios (RRs) or odds ratio (ORs) and their 95% confidence intervals were calculated and pooled through the STATA 12.0. A total of 6 studies were finally selected in the meta-analysis. No significant association was found between total anthocyanins consumption and gastric cancer risk (RR=0.92, 95%CI: 0.81-1.04). Likewise, there was also no significant evidence of the relationship between anthocyanins intake and gastric cancer in tumor site (cardia: RR=0.90, 95%CI: 0.62-1.31; noncardia: RR=0.86, 95%CI: 0.69-1.07) and gender (men: RR=1.02, 95%CI: 0.73-1.40; women: RR=0.80, 95%CI: 0.52-1.23). The dose-response relationship was also not found in this meta-analysis. The Grades of Recommendations Assessment, Development and Evaluation (GRADE) quality in our study was very low. The results of our meta-analysis suggested the intake of anthocyanins had no association with the risk of gastric cancer and further studies are needed.

Anthocyanin Consumption and Risk of Colorectal Cancer: A Meta-Analysis of Observational Studies.

This meta-analysis aimed to summarize the association between anthocyanin consumption and the risk of colorectal cancer. All relative articles were located on online databases, including PubMed, Embase, and the Cochrane Library as of June 11, 2018. Risk ratios (RRs) or odds ratio and their 95% confidence intervals (CIs) were calculated through the STATA 12.0 software package. A total of seven studies were included in the meta-analysis. A significant inverse association was found between total anthocyanin consumption and colorectal cancer risk (RR = 0.78; 95% CI, 0.64-0.95). Likewise, there was significant evidence of a relationship between anthocyanin intake and colorectal cancer in the colon site (RR = 0.81; 95% CI, 0.71-0.92); men (RR = 0.88; 95% CI, 0.81-0.95), and case-control studies (RR = 0.69; 95% CI, 0.60-0.78). A dose-response relationship was not found in this meta-analysis. The Grades of Recommendations Assessment, Development, and Evaluation quality in our study was very low. This meta-analysis indicates that anthocyanin consumption is inversely associated with the risk of colorectal cancer. Anthocyanins may play an active role in the prevention of colorectal cancer. Key teaching points: Some epidemiological studies found an inverse correlation between the high consumption of anthocyanins and low risk of colorectal cancer. Because of this structure, anthocyanins/anthocyanidins have a powerful capability of donating electrons, which can be characterized as antioxidant properties. Anthocyanins can also inhibit colon cancer by interfering in the cell cycle and inducing the effect of anti-proliferation and apoptosis. The formation of cytoplasmic vacuoles in cells also indicates that anthocyanins may induce autophagy. From the findings of nonrandomized controlled trials, anthocyanins may play an active role in the prevention of colorectal cancer.

Biochar reduces cadmium accumulation in rice grains in a tungsten mining area-field experiment: effects of biochar type and dosage, rice variety, and pollution level.

Cadmium (Cd)-contaminated rice (Oryza sativa) in Southern China is a great threat to food security, and the paddy soil remediation is urgently needed to reduce Cd accumulation in rice. Application of biochar could effectively immobilize soil Cd and reduce Cd uptake by rice. Fields that were applied with soil treatments including control and 15 and 30 t ha-1 each hickory nut shell-derived biochar (KC) or maize straw-derived biochar (MC), and grown with two rice varieties (hybrid rice and late japonica rice) were selected for this study. The long-term effect of biochars on decreasing Cd bioavailability in paddy soils was evaluated. The results showed when MC was applied at 15 t ha-1, DTPA-Cd (soil cadmium extracted by diethylenetriamine pentaacetic acid) was reduced by 20.0 and 34.5% in Field A (slightly Cd pollution) and B (moderately Cd pollution), respectively. In Field B, soil DTPA-Cd concentrations with application of 30 t ha-1 biochars were all lower than that of 15 t ha-1 biochar, but there were no significant differences between the two types of biochars. Cd concentration in rice grains and straws of hybrid rice are two times more than those of late japonica rice. Cd bio-concentration factor both of grains and straw was significantly increased by biochar application, which in Field A was higher than that in Field B. Our results suggest that biochars reduce Cd accumulation in rice grains by immobilizing soil Cd. KC has a higher potential in lowering Cd bioavailability than MC. Hybrid rice should be prohibited to cultivate in these areas.

MK-8325: A silyl proline-containing NS5A inhibitor with pan-genotype activity for treatment of HCV.

HCV NS5A inhibitors have shown impressive in vitro potency profiles in HCV replicon assays thus making them attractive components for inclusion in an all oral fixed dose combination regimen. Herein, we describe the discovery and characterization of silyl proline-containing HCV NS5A inhibitor MK-8325 with good pan-genotype activity and acceptable pharmacokinetic properties.

Development of a prodrug of hydantoin based TACE inhibitor.

Our research on hydantoin based TNF-α converting enzyme (TACE) inhibitors led to fused bi-heteroaryl hydantoin series that demonstrate sub-nanomolar potency (Ki) as well as excellent activity in human whole blood (hWBA). However, lead compound 2 posed some formulation challenges which prevented it for further development. A prodrug approach was investigated to address this issue. The pivalate prodrug 3 can be formulated as stable neutral form and demonstrated improved DMPK properties when compared with parent compound.

Structure-activity relationships of proline modifications around the tetracyclic-indole class of NS5A inhibitors.

We describe the impact of proline modifications, in our tetracyclic-indole based series of nonstructural protein 5A (NS5A) inhibitors, to their replicon profiles. This work identified NS5A inhibitors with an improved and flattened resistance profiles.

Matched and mixed cap derivatives in the tetracyclic indole class of HCV NS5A inhibitors.

A matched and mixed capping SAR study was conducted on the tetracyclic indole class of HCV NS5A inhibitors to examine the influence of modifications of this region on the overall HCV virologic resistance profiles.

Alkyl substituted aminal derivatives of HCV NS5A inhibitor MK-8742.

HCV NS5A inhibitors have demonstrated impressive in vitro potency profiles in HCV replicon assays and robust HCV RNA titer reduction in the clinic making them attractive components for inclusion in an all oral fixed dose combination regimen for the treatment of HCV infection. Herein we describe our continued research efforts around the alkyl "Z group" modification of the tetracyclic indole-based NS5A inhibitor MK-8742, which led to the discovery of a series of potent NS5A inhibitors. Compounds 10 and 19 are of particular interests since they are as potent as our previous leads and have much improved rat pharmacokinetic profiles.

Discovery of fused tricyclic core containing HCV NS5A inhibitors with pan-genotype activity.

HCV NS5A inhibitors have demonstrated impressive in vitro potency profiles in HCV replicon assays and robust HCV RNA titer reduction in the clinic making them attractive components for inclusion in an all oral fixed dose combination regimen for the treatment of HCV infection. Herein, we describe research efforts that led to the discovery of a series of fused tricyclic core containing HCV NS5A inhibitors such as 24, 39, 40, 43, and 44 which have pan-genotype activity and are orally bioavailable in the rat.

Discovery of silyl proline containing HCV NS5A inhibitors with pan-genotype activity: SAR development.

HCV NS5A inhibitors have shown impressive in vitro potency profiles in HCV replicon assays thus making them attractive components for inclusion in an all oral fixed dose combination treatment regimen. Herein we describe the research efforts that led to the discovery of silyl proline containing HCV NS5A inhibitors such as 7e and 8a with pan-genotype activity profile and acceptable pharmacokinetic properties.