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.

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.

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.

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.

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.

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.

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.

Roles of oxidative damage and mitochondria-mediated apoptosis in ethylbenzene-induced hepatotoxic effects in rat.

The mechanisms underlying hepatoxic effects of ethylbenzene still remain unknown. We investigated the toxic effects of ethylbenzene on liver and explored the mechanism of mitochondria-mediated apoptosis pathway. Forty male Sprague-Dawley rats were used as an in vivo model with ethylbenzene inhalation of 0, 433.5 mg/m(3), 4335 mg/m(3) and 6500 mg/m(3) for 13 weeks. Levels of malondialdehyde, glutathione, glutathione peroxidase and superoxide dismutase were assayed. Meanwhile, the ultrastructure of hepatic tissues was observed and cell apoptosis was detected via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Furthermore, we investigated the expression levels of mRNA and protein of bax, bcl-2, cytochrome c, caspase-9 and caspase-3 in rat liver tissues. Compared with control group, the malondialdehyde levels were significantly elevated while glutathione levels and activities of glutathione peroxidase and superoxide dismutase were decreased, respectively. The mitochondria of liver appeared swollen with vacuolar structure and loss of cristae in 6500 mg/m(3) ethylbenzene-treated group, and ethylbenzene induced a significant increase in the percentage of apoptotic cells as compared to the control group. In addition, enhanced mRNA and protein expression levels of all measured genes were observed in ethylbenzene-treated groups except the decreased bcl-2 expression levels. Our results indicated that ethylbenzene may induce oxidative damage and apoptosis in rat liver. Mitochondrial-mediated pathway was involved in the apoptosis process.

[Effects of 1-bromopropane on liver and kidney functions of exposed workers].

OBJECTIVE: To study the effects of 1-bromopropane (1-BP) on liver and kidney functions of exposed workers. METHODS: Occupational health situation in three 1-BP plants was investigated. Fifty-four workers from the 1-BP manufacturing line were chose to be contact group, while 42 workers from non-1-BP manufacturing line as control group. All workers underwent questionnaire survey, liver function test as well as kidney function test. RESULT: Working years has no impact on liver and kidney functions of workers from contact group. Compared with the control, liver and kidney functions test of the two groups showed no statistical difference either. CONCLUSION: The present investigation doesn't prove any impact of occupational 1-BP exposure on worker's liver and kidney functions.

IV. Discovery of CXCR3 antagonists substituted with heterocycles as amide surrogates: improved PK, hERG and metabolic profiles.

The structure-human CXCR3 binding affinity relationship of a series of pyridyl/pyrazinyl-piperazinyl-piperidine derivatives were explored with a focus to improve PK, hERG and metabolic profiles. Several small heterocycles were identified as amide surrogates, which minimized many potential metabolite issues. During the course of SAR development, we have observed the additive effect of desirable functional groups to improve hERG and PK profiles which lead to the discovery of many clinically developable CXCR3 antagonists with excellent overall profile.

[Effect of smoking on the microRNAs expression in pneumoconiosis patients].

OBJECTIVE: To investigate the effect of smoking on the microRNAs (miRNAs) expression in pneumoconiosis patients. METHODS: Real-time qPCR was used to measure the expression levels of miR-21, miR-200c, miR-16, miR-204, miR-206, miR-155, let-7g, miR-30b, and miR-192 in 36 non-smoking patients with pneumoconiosis and 38 smoking patients with pneumoconiosis, and the differences in expression levels between the two groups were evaluated by two-independent samples t-test. RESULTS: The expression of miR-192 in serum showed a significant difference between non-smoking and smoking pneumoconiosis patients (P < 0.05), and it decreased gradually in smoking patients with stage I and II pneumoconiosis. In the serum of all pneumoconiosis patients, the expression level of miR-16 was the highest, while the expression level of miR-204 was the lowest. CONCLUSION: Pneumoconiosis patients have differential expression of miRNAs in serum, and smoking has an effect on the miRNAs expression in pneumoconiosis patients.

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.

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.

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.

[The inhibitory effect of latent transforming growth factor ß1 activation and silicosis by CD36 targeted RNA interference in silicosis model of rat].

OBJECTIVE: To investigate the inhibitory effects of CD36-targeting RNA interference on the latent transforming growth factor ß1 (L-TGF-ß1) activation and silicotic fibrosis in rat silicosis model. METHODS: Wistar rats were divided into four groups: saline control group (n=24), SiO2 model group (10 mg SiO2 per rat) (n=24), SiO2+Lv-shCD36 group (lentiviral vector expressing specific shRNA against CD36) (n=24), and SiO2+Lv-shCD36-NC group (non-silence control lentivirus) (n=24). At 7, 21, and 28 d after instillation, the rats were sacrificed. The activity of TGF-ß1 in bronchoalveolar lavage fluid (BALF) was measured by evaluating its inhibitory effect on the proliferation of mink lung epithelial cells. The pathological changes of lung tissue were observed by HE staining and van Gieson staining. The hydroxyproline content in the lungs was determined by alkaline lysis method. RESULTS: At 7 d after instillation, the expression of CD36 mRNA in alveolar macrophages was significantly lower in the SiO2+Lv-shCD36 group than in the saline control group, SiO2 model group, and SiO2+Lv-shCD36-NC group (P < 0.05); the quantity and percentage of active TGF-ß1 in BALF were significantly lower in the SiO2+Lv-shCD36 group than in the SiO2 model group and SiO2+Lv-shCD36-NC group (P < 0.05). At 28 d after instillation, there were cellular silicotic nodules in the lungs of rats in SiO2+Lv-shCD36 group and fibrotic cellular silicotic nodules in the lungs of rats in SiO2 model group and SiO2+Lv-shCD36-NC group. At 21 and 28 d after instillation, the hydroxyproline content was significantly lower in the SiO2+Lv-shCD36 group than in the SiO2 model group and SiO2+Lv-sh CD36-NC group (P < 0.05). CONCLUSION: CD36-targeting RNA interference has inhibitory effects on the L-TGF-ß1 activation and silicotic fibrosis in rat silicosis model.