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Objective: To explore the correlation between abnormal expression of serum 25(OH)D and immune-related factors and subclinical atherosclerosis (AS) in patients with impaired glucose regulation. Methods: A total of 142 patients with impaired glucose regulation admitted to Zaozhuang Municipal Hospital from December 2019 to April 2021 were selected. The patients with impaired glucose regulation were selected as the control group (n=86) and patients with subclinical atherosclerosis were selected as the observation group (n=56) according to carotid ultrasound examination and bramolecular pulse wave conduction velocity. The serum 25(OH)D and immune factors were compared between the two groups. Carotid ultrasound was used to measure the carotid inti-media thickness, and Pearson method was used to determine the correlation between the intimedia thickness, 25(OH)D and immune factors. Baseline data and hematological indicators were collected, univariate and multivariate Logistic regression analysis was performed to determine the influencing factors of subclinical atherosclerosis, and ROC curve was used to evaluate the diagnostic efficacy of serum 25(OH)D and immune factors in subclinical atherosclerosis. Results: The serum 25(OH)D [ (24.01 ±4.87) mmol/L vs (30.74±5.01)mmol/L, t=7.909, P=0.000) in the observation group was significantly lower than that of control group. TNF-α [(48.32± 8.02)ng/Lr vs (33.21±9.00)ng/L, t=10.199, P=0.000) and IL-6 [(41.22±9.43)ng/L vs (30.21±7.01)ng/L, t=7.492, P=0.000) in observation group were significantly higher than those in control group. Carotid intima-media thickness was negatively correlated with serum 25(OH)D (r=-0.428, P<0.001), and was positively correlated with serum TNF-α and IL-6 (r=0.574, 0.577, P<0.001). Logistic regression analysis showed that serum 25(OH)D (OR=0.520, 95%CI: 0.401~0.675), serum TNF-α (OR=1.667, 95%CI: 1.131~2.457) and serum IL-6 (OR=1.478, 95%CI: 1.213~1.802) were the influencing factors of subclinical atherosclerosis. ROC curve showed that the optimal cut-off value of serum 25 (OH) D was 28.32 mmol/L, and the corresponding sensitivity, specificity and AUC were 69.64%, 70.93% and 0.803 (95%CI: 0.749~0.855), respectively. The critical value of serum TNF-α was 40.56 ng/L, corresponding sensitivity was 71.43%, specificity was 72.09%, and AUC was 0.761 (95%CI: 0.717~0.823). Serum IL-6 cut-off value was 36.13 ng/L, corresponding sensitivity was 60.71%, specificity was 60.47%, and AUC was 0.627 (95%CI: 0.566~0.702). The sensitivity, specificity and AUC of regression analysis were 85.71%, 81.40% and 0.889(95%CI: 0.830~0.915). Conclusion: Serum 25 (OH)D and immune-related factors alone and in combination can effectively predict the occurrence of subclinical atherosclerosis, which is correlated with carotid intima-media thickness, and serum 25 (OH) D and immune-related factors are predictors of subclinical atherosclerosis. [ABSTRACT FROM AUTHOR]

Trichomonas gallinae (T. gallinae) is a flagellated protozoan and the causative agent of trichomoniasis, or canker, in birds. In the current study, the prevalence of T. gallinae was firstly investigated in five breeds. According to the results of the prevalence study, White King pigeons were selected as the experimental animals. A total of 135 White King squabs at one day of age were randomly divided into two groups and raised in separate isolators. The challenged group (N = 100) was challenged intranasally with 5 × 106 parasites/mL of the T. gallinae strain, and the control group (N = 35) was intranasally administered medium of equivalent volume. At 1, 2, 3 and 5 days post infection (DPIs), the crops and esophagi were collected for RNA extraction and formaldehyde fixation. The results showed that prevalence of T. gallinae in the five breeds ranged from 27.13% (White Carneau) to 43.14% (White King). After the challenge, mild microscopic lesions were observed in both tissues. Apoptosis rates were higher in the challenged group than in the control group at 2 and 5 DPIs in the crop and at 1, 2 and 7 DPIs in the esophagus. For both tissues, relative expression of IL-1β increased dramatically at the beginning and decreased at 5 DPIs, and TGF-β increased stably in the challenged group.

This experiment aimed to study the effects on liver tissue structure and immune performance of grass carp Ctenopharyngodon idellus when the common reed Phragmites australis is in its diet, instead of wheat Triticum aestivum L. Seventy-two healthy grass carps (145.52 ± 2.56 g) were randomly divided into three groups according to their body weight. Fishes in each group were fed an essential diet with 0% (control group), 50% and 100% (test group) common reed, instead of wheat, respectively. After feeding for 41 days, the changes of serum biochemical indices, liver tissue structure and immune related indices of grass carp were detected. The results showed that, compared with the control group, the activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) in the substitution groups were significantly increased (p < 0.05), but still at a normal level. The contents of total protein, albumin and globulin did not change significantly (p > 0.05). Compared with the control group, the liver cells of grass carp in the substitution groups had clear boundaries, tight arrangement and less vacuolation. The contents of serum interleukin-1 (IL-1) and complement 3 (C3) in the 100% substitution group were significantly higher than those in the control group (p < 0.05), and the contents of liver IL-1 and total complement (CH50) in the 100% substitution group were significantly higher than those in the control group (p < 0.05). The contents of IL-1, C3 in serum and IL-1, CH50 in liver in the 50% substitution group were significantly higher than those in the control group (p < 0.05). The mRNA relative expression levels of C3, IL-1, MHC-I and interferon (IFN) in the head-kidney, kidney, liver and spleen of grass carp were significantly affected by feeding the grass carp with different common reed substitution ratios (p < 0.05). In summary, common reed, instead of wheat, in feed can improve the liver tissue structure, and increase the non-specific immune response level, of grass carp. [ABSTRACT FROM AUTHOR]

Background: Growing evidence suggests that colorectal cancer (CRC) should be considered a heterogeneous disease. The right side (RCC) and left side (LCC) colorectal cancer have different clinical characteristics and immune landscapes. The aim of this study was to analyze differential expression and prognostic correlation of immune-related factors between RCC and LCC. Methods: The gene expression profile and clinical characteristics of CRC patients were retrieved from The Cancer Genome Atlas data portal (n=525). Using a deconvolution algorithm, immune cell infiltration in RCC and LCC based on the RNA-seq data was analyzed. Differentially expressed genes (DEGs) were obtained by performing differential gene expression analysis. Immune-related DEGs were derived by the intersection with immune-related factors downloaded from the IMMPORT database. To further validate the findings, we applied immunohistochemical (IHC) staining of a CRC tissue microarray (TMA). The distribution of immune cells in RCC and LCC and changes in the expression of immune molecules on their membranes were verified. The expression levels of circulating cytokines were measured by flow cytometry to detect the cytokines secreted by immune cells in RCC and LCC. Furthermore, to reveal the prognostic value of differential immune factors on RCC and LCC patients, survival analysis based on mRNA levels using TCGA cohort and survival analysis using protein levels was performed using our CRC patients. Results: The infiltration of immune cells differed between RCC and LCC, the infiltration degree of macrophages M0 was significantly higher in LCC, while the infiltration degree of differentiated macrophages M1 and M2, CD4+ T and CD8+ T cells was significantly higher in RCC. The expression of related molecules by immune cells also differed between RCC and LCC. The expression of 7 genes in RCC was higher than that in LCC, which were CCR5, CD209, CD8A, HCK, HLA-DPB1, HLA-DQA1, HLA-DRA, respectively. Meanwhile, the expression of 2 genes in LCC was higher than in RCC, which were IL34 and PROCR. Patients with RCC having high expression of HLA-DQA1 mRNA or proteins had better survival and LCC patients with high expression of IL 34 mRNA or protein had better survival. Conclusions: In this study, we comprehensively compared differences in immune cells and regulating factors between left and right colorectal cancer. Different expression patterns and their effects on survival were identified. The analysis of immune-related factors may provide a theoretical basis for precise immunotherapy of RCC and LCC. [ABSTRACT FROM AUTHOR]

Background: Previous studies have shown that EDCs may activate nuclear transcription factor, such as activator protein-1 (AP-1), nuclear factor of activated Tcells (NF-AT) and nuclear factor kappa B (NF-κB) in the process of immune damage. At the same time, some experts believed that estrogen may play an important role in this process. As a typical representative of EDCs, nonylphenol (NP) has not been reported. The aim of this work was to explore the relationship between the immune inflammatory damage and the changes in estrogen expression in male rats during the chronic exposure to NP at environmental concentrations. Sixty SPF Sprague–Dawley rats were divided into five groups (n = 12 per group): blank control group (corn oil), low-dose NP exposure group (0.4 mg/kg/d), medium-dose NP exposure group (4 mg/kg/d), high-dose NP exposure group (40 mg/kg/d), and estradiol control group (E2: 30 μg/kg/d). Results: Compared with the control group, rat spleen organ coefficient, number of spleen nodules, relative area of lymph nodes and white pulp were relatively reduced in the L (NP, 0.4 mg/kg) and H (NP, 40 mg/kg) exposure dose groups (P < 0.001). Lymphocytes were rich in cytoplasm, mitochondria were swollen, part of the cristae was reduced, and rough endoplasmic reticulum was expanded. The serum levels of IgG (P < 0.001) and IgM (P = 0.002) showed a downward trend. The percentage of Th cells (CD3+CD4+) was significantly decreased (P < 0.001), and the percentage of B lymphocytes shows an opposite trend (P < 0.001). Giemsa staining showed that the number of neutrophils (P < 0.001) was increased. The expressions of estrogen receptor ER-α and ER-β protein in the spleen increased significantly (P < 0.001). The expressions of AP-1 protein and NF-AT protein in the spleen were increased, and the expression of NF-KB protein was decreased (P < 0.001). The expressions of IL-4, ER-α and ER-β (P < 0.001) levels in serum increased. The mRNA-seq bioinformatics detection showed the final differentially expressed immune-inflammatory-related genes between the control and H-NP groups as follow: down-regulated: TLR4, Gata3, IL12, up-regulated: TNF-a, IL10, INOS. The mRNA expressions of ER-α, ER-β, NF-KB, IL4, AP-1, TLR4, Gata3, and NF-AT were consistent with the results of mRNA-seq analysis. NP content was correlated with the expressions of ER-α, ER-β, IL4, AP-1, NF-AT, TLR4, NF-KB, as well as IL-12 proteins in the spleen tissue ([r] < 1, P < 0.05). Conclusions: Chronic exposure to NP at environmental concentration could cause immune dysfunction, resulting in immunotoxicity and inflammatory effects, and lead to changes in the activity of transcription factors and differential immune inflammatory factors in rats. [ABSTRACT FROM AUTHOR]

This experiment aimed to study the effects on liver tissue structure and immune performance of grass carp Ctenopharyngodon idellus when the common reed Phragmites australis is in its diet, instead of wheat Triticum aestivum L. Seventy-two healthy grass carps (145.52 ± 2.56 g) were randomly divided into three groups according to their body weight. Fishes in each group were fed an essential diet with 0% (control group), 50% and 100% (test group) common reed, instead of wheat, respectively. After feeding for 41 days, the changes of serum biochemical indices, liver tissue structure and immune related indices of grass carp were detected. The results showed that, compared with the control group, the activities of serum alanine transaminase (ALT) and aspartate transaminase (AST) in the substitution groups were significantly increased (p < 0.05), but still at a normal level. The contents of total protein, albumin and globulin did not change significantly (p > 0.05). Compared with the control group, the liver cells of grass carp in the substitution groups had clear boundaries, tight arrangement and less vacuolation. The contents of serum interleukin-1 (IL-1) and complement 3 (C3) in the 100% substitution group were significantly higher than those in the control group (p < 0.05), and the contents of liver IL-1 and total complement (CH50) in the 100% substitution group were significantly higher than those in the control group (p < 0.05). The contents of IL-1, C3 in serum and IL-1, CH50 in liver in the 50% substitution group were significantly higher than those in the control group (p < 0.05). The mRNA relative expression levels of C3, IL-1, MHC-I and interferon (IFN) in the head-kidney, kidney, liver and spleen of grass carp were significantly affected by feeding the grass carp with different common reed substitution ratios (p < 0.05). In summary, common reed, instead of wheat, in feed can improve the liver tissue structure, and increase the non-specific immune response level, of grass carp.

Trichomonas gallinae ( T. gallinae ) is a flagellated protozoan and the causative agent of trichomoniasis, or canker, in birds. In the current study, the prevalence of T. gallinae was firstly investigated in five breeds. According to the results of the prevalence study, White King pigeons were selected as the experimental animals. A total of 135 White King squabs at one day of age were randomly divided into two groups and raised in separate isolators. The challenged group (N = 100) was challenged intranasally with 5 × 10 6 parasites/mL of the T. gallinae strain, and the control group (N = 35) was intranasally administered medium of equivalent volume. At 1, 2, 3 and 5 days post infection (DPIs), the crops and esophagi were collected for RNA extraction and formaldehyde fixation. The results showed that prevalence of T. gallinae in the five breeds ranged from 27.13% (White Carneau) to 43.14% (White King). After the challenge, mild microscopic lesions were observed in both tissues. Apoptosis rates were higher in the challenged group than in the control group at 2 and 5 DPIs in the crop and at 1, 2 and 7 DPIs in the esophagus. For both tissues, relative expression of IL-1β increased dramatically at the beginning and decreased at 5 DPIs, and TGF-β increased stably in the challenged group.

A representative echinoderm, the sea cucumber Apostichopus japonicus has a special regeneration mechanism. The sea cucumber has no specific immune tissues or organs. Coelomocytes of sea cucumbers are involved in their cellular and humoral immunity. In this study, expression profiles of the main immune-related factors in sea cucumber coelomocytes were detected during coelomocyte regeneration after evisceration in A. japonicus. Immune-related factors Cu/Zn superoxide dismutase (Cu/Zn SOD), Catalase (CAT), C-type lysozyme (C-LYZ), Cathepsin D (CTSD), Melanotransferrin (MTF), Toll-like receptor (TLR), C-type lectin (C-LCT), Complement 3 (C3), Myeloid differentiation factor 88 (MYD88), Nuclear factor kappa-B (NF-κB), NF-κB inhibitor (IKB), TNF receptor-associated factor 6 (TRAF6), Peroxiredoxins (PRX), Nitric oxide synthase(NOS), Caspase-2 (CASP-2), Phenoloxidase (PO), and Glutathione Peroxidase(GPX), Heat shock protein 70 (HSP70) were detected by real-time fluorescent quantitative PCR at different time points during regeneration. The main immune-related genes in sea cucumber coelomocytes were significantly differentially expressed after evisceration, and an upregulation was observed for the majority of the considered genes. In summary, the discharge of viscera had a significant effect on expression of immune-related genes of sea cucumber coelomocytes. The expression level of each gene had a certain correlation with the sea cucumber regeneration process. The results provide reference data for the immune response of coelomocytes during regeneration.

The 4-week feeding experiments were conducted to investigate the effects of astragalus polysaccharides (APS) on hemocytes phagocytosis and gene expression of immune-related factors in Eriocheir sinensis—with initial weights of 6.11 ± 2.25 g. The phagocytic activity of hemocytes in E. sinensis was determined by fluorescence labeling, and the expression of immune-related factor genes in different tissues of E. sinensis was analyzed by quantitative real-time PCR (qPCR). Results showed that APS significantly improved the phagocytic activity of hemocytes in E. sinensis (p < 0.05), and the phagocytic activity of hemocytes in the 0.1% APS group was the highest (50.84%). Moreover, 0.05% APS significantly increased the expression of glutathione peroxidase (GSH-Px), anti-lipopolysaccharide (ALF), phenol oxidase (PO) and heat shock protein (HSP) genes in the hemocytes, and HSP in the hepatopancreas, and masquerade-like protein (MasL) in the gill (p < 0.05). Furthermore, 0.1% APS significantly increased the expression levels of PO, GSH-Px, ALF, MasL, and HSP genes in the hemocytes and gill (p < 0.05) and significantly increased the expression levels of PO, Crusl, and HSP genes in crab hepatopancreas (p < 0.05). In addition, 0.2% APS significantly increased the expression levels of all immune-related factor genes in the hemocytes and gill (p < 0.05) and significantly increased the expression levels of PO, ALF, Crusl, and HSP genes in the crab hepatopancreas (p < 0.05). It is estimated that APS could enhance the immune function of E. sinensis by increasing the phagocytic activity of hemocytes and the expression levels of the PO, GSH-Px, ALF, Crus1, MasL, and HSP genes. [ABSTRACT FROM AUTHOR]

A representative echinoderm, the sea cucumber Apostichopus japonicus has a special regeneration mechanism. The sea cucumber has no specific immune tissues or organs. Coelomocytes of sea cucumbers are involved in their cellular and humoral immunity. In this study, expression profiles of the main immune-related factors in sea cucumber coelomocytes were detected during coelomocyte regeneration after evisceration in A. japonicus. Immune-related factors Cu/Zn superoxide dismutase (Cu/Zn SOD), Catalase (CAT), C-type lysozyme (C-LYZ), Cathepsin D (CTSD), Melanotransferrin (MTF), Toll-like receptor (TLR), C-type lectin (C-LCT), Complement 3 (C3), Myeloid differentiation factor 88 (MYD88), Nuclear factor kappa-B (NF-κB), NF-κB inhibitor (IKB), TNF receptor-associated factor 6 (TRAF6), Peroxiredoxins (PRX), Nitric oxide synthase(NOS), Caspase-2 (CASP-2), Phenoloxidase (PO), and Glutathione Peroxidase(GPX), Heat shock protein 70 (HSP70) were detected by real-time fluorescent quantitative PCR at different time points during regeneration. The main immune-related genes in sea cucumber coelomocytes were significantly differentially expressed after evisceration, and an upregulation was observed for the majority of the considered genes. In summary, the discharge of viscera had a significant effect on expression of immune-related genes of sea cucumber coelomocytes. The expression level of each gene had a certain correlation with the sea cucumber regeneration process. The results provide reference data for the immune response of coelomocytes during regeneration. [ABSTRACT FROM AUTHOR]

The present study evaluated the effects of dietary β-glucan (0, 0.05%, 0.1%, and 0.2%) on growth performance after 42 days of feeding. Thereafter, rainbow trout ( Oncorhynchus mykiss ) were infected with Aeromonas salmonicida , and survival rates as well as the regulating processes of stress- and immune-related factors were analyzed. In general, higher dietary β-glucan levels obviously improved specific growth rate (SGR), weight gain (WG) and feed efficiency (FE) ( P ≤ 0.05). Survival rates in β-glucan groups increased significantly compared with the control group after A. salmonicida infection ( P ≤ 0.05). Serum total superoxide dimutase (T-SOD), peroxidase (POD) as well as catalase (CAT) activities, and their mRNA expressions in the head kidney of fish in the β-glucan groups generally increased to higher levels after infection, and more quickly, compared with in the control group. Serum lysozyme (LSZ) and its expression in the head kidney in β-glucan groups reached a higher peak earlier than in the control group. Serum glutamic oxalacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels in the β-glucan groups were significantly lower than in the control group ( P ≤ 0.05). The peak of heat shock protein 70 (HSP70) expression in the 0.2% β-glucan group was higher and occurred earlier than in other groups ( P ≤ 0.05). These results confirm that 0.1% and 0.2% dietary β-glucan are beneficial for promoting growth in rainbow trout and enhancing resistance against A. salmonicida . Furthermore, β-glucan could play an important role in regulating stress- and immune-related factors in rainbow trout to more quickly fight against bacterial infection. [ABSTRACT FROM AUTHOR]

Background: Growing evidence suggests that colorectal cancer (CRC) should be considered a heterogeneous disease. The right side (RCC) and left side (LCC) colorectal cancer have different clinical characteristics and immune landscapes. The aim of this study was to analyze differential expression and prognostic correlation of immune-related factors between RCC and LCC., Methods: The gene expression profile and clinical characteristics of CRC patients were retrieved from The Cancer Genome Atlas data portal (n=525). Using a deconvolution algorithm, immune cell infiltration in RCC and LCC based on the RNA-seq data was analyzed. Differentially expressed genes (DEGs) were obtained by performing differential gene expression analysis. Immune-related DEGs were derived by the intersection with immune-related factors downloaded from the IMMPORT database. To further validate the findings, we applied immunohistochemical (IHC) staining of a CRC tissue microarray (TMA). The distribution of immune cells in RCC and LCC and changes in the expression of immune molecules on their membranes were verified. The expression levels of circulating cytokines were measured by flow cytometry to detect the cytokines secreted by immune cells in RCC and LCC. Furthermore, to reveal the prognostic value of differential immune factors on RCC and LCC patients, survival analysis based on mRNA levels using TCGA cohort and survival analysis using protein levels was performed using our CRC patients., Results: The infiltration of immune cells differed between RCC and LCC, the infiltration degree of macrophages M0 was significantly higher in LCC, while the infiltration degree of differentiated macrophages M1 and M2, CD4+ T and CD8+ T cells was significantly higher in RCC. The expression of related molecules by immune cells also differed between RCC and LCC. The expression of 7 genes in RCC was higher than that in LCC, which were CCR5, CD209, CD8A, HCK, HLA-DPB1, HLA-DQA1, HLA-DRA, respectively. Meanwhile, the expression of 2 genes in LCC was higher than in RCC, which were IL-34 and PROCR. Patients with RCC having high expression of HLA-DQA1 mRNA or proteins had better survival and LCC patients with high expression of IL 34 mRNA or protein had better survival., Conclusions: In this study, we comprehensively compared differences in immune cells and regulating factors between left and right colorectal cancer. Different expression patterns and their effects on survival were identified. The analysis of immune-related factors may provide a theoretical basis for precise immunotherapy of RCC and LCC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hu, Ding, Wu, Gao, Ge, Shao, Liu and Ye.)