Effects of metformin, saxagliptin and repaglinide on gut microbiota in high-fat diet/streptozocin-induced type 2 diabetic mice.

INTRODUCTION: There has been increasing evidence that the gut microbiota is closely related to type 2 diabetes (T2D). Metformin (Met) is often used in combination with saxagliptin (Sax) and repaglinide (Rep) for the treatment of T2D. However, little is known about the effects of these combination agents on gut microbiota in T2D. RESEARCH DESIGN AND METHODS: A T2D mouse model induced by a high-fat diet (HFD) and streptozotocin (STZ) was employed. The T2D mice were randomly divided into six groups, including sham, Met, Sax, Rep, Met+Sax and Met+Rep, for 4 weeks. Fasting blood glucose level, serum biochemical index, H&E staining of liver, Oil red O staining of liver and microbiota analysis by 16s sequencing were used to access the microbiota in the fecal samples. RESULTS: These antidiabetics effectively prevented the development of HFD/STZ-induced high blood glucose, and the combination treatment had a better effect in inhibiting lipid accumulation. All these dosing regimens restored the decreasing ratio of the phylum Bacteroidetes: Firmicutes, and increasing abundance of phylum Desulfobacterota, expect for Met. At the genus level, the antidiabetics restored the decreasing abundance of Muribaculaceae in T2D mice, but when Met was combined with Rep or Sax, the abundance of Muribaculaceae was decreased. The combined treatment could restore the reduced abundance of Prevotellaceae_UCG-001, while Met monotherapy had no such effect. In addition, the reduced Lachnospiraceae_NK4A136_group was well restored in the combination treatment groups, and the effect was much greater than that in the corresponding monotherapy group. Therefore, these dosing regimens exerted different effects on the composition of gut microbiota, which might be associated with the effect on T2D. CONCLUSIONS: Supplementation with specific probiotics may further improve the hypoglycemic effects of antidiabetics and be helpful for the development of new therapeutic drugs for T2D.

The transcriptome sequencing analysis reveals immune mechanisms of soybean fermented powder on the loach (Misgurnus anguillicaudatus) in response to Lipopolysaccharide (LPS) infection.

The loach (Misgurnus anguillicaudatus), a small commercial fish that is widely cultivated for its high-quality protein, vitamins, minerals, and essential amino acid, is a member of the genus Misgurnus and the family Cyprinidae. In this study, we gave the LPS-injected loach fermented soybean meal and used transcriptome sequencing to investigate the impact of the fermented soybean powder on the loach's immune system. 3384 up-regulated genes and 12116 down-regulated genes were found among the 15500 differentially expressed genes, according to the results. The differentially expressed genes were shown to be involved in cellular processes, metabolic processes, cellular anatomical entities, and binding, according to the Go functional annotation. Meanwhile, the KEGG enrichment analysis indicated that the soybean fermented powder treated groups showed significant differences in DNA replication, Nucleotide excision repair, Fanconi anemia pathway, and Base excision repair pathways, suggesting that these pathways are closely related to the enhancement of the immune function of loach by soybean fermented powder. The particular conclusions not exclusively can provide a new conception for the rational utilization of soybean fermented powder but also can provide theoretical guidance for the subsequent healthy breeding of loach.

A humanized 4-1BB-targeting agonistic antibody exerts potent antitumor activity in colorectal cancer without systemic toxicity.

BACKGROUND: Colorectal cancer (CRC) is one of the most common malignancies and the patient survival rate remains unacceptably low. The anti-programmed cell death-1 (PD-1)/programmed cell death ligand 1 (PD-L1) antibody-based immune checkpoint inhibitors have been added to CRC treatment regimens, however, only a fraction of patients benefits. As an important co-stimulatory molecule, 4-1BB/CD137 is mainly expressed on the surface of immune cells including T and natural killer (NK) cells. Several agonistic molecules targeting 4-1BB have been clinically unsuccessful due to systemic toxicity or weak antitumor effects. We generated a humanized anti-4-1BB IgG4 antibody, HuB6, directed against a unique epitope and hypothesized that it would promote antitumor immunity with high safety. METHODS: The antigen binding specificity, affinity and activity of HuB6 were determined by enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), biolayer interferometry (BLI) and flow cytometry. The antitumor effects were evaluated in humanized mice bearing syngeneic tumors, and possible toxicity was evaluated in humanized mice and cynomolgus monkeys. RESULTS: HuB6 showed high specificity and affinity for a binding epitope distinct from those of other known 4-1BB agonists, including utomilumab and urelumab, and induced CD8 + T, CD4 + T and NK cell stimulation dependent on Fcγ receptor (FcγR) crosslinking. HuB6 inhibited CRC tumor growth in a dose-dependent manner, and the antitumor effect was similar with urelumab and utomilumab in humanized mouse models of syngeneic CRC. Furthermore, HuB6 combined with an anti-PD-L1 antibody significantly inhibited CRC growth in vivo. Additionally, HuB6 induced antitumor immune memory in tumor model mice rechallenged with 4 × 106 tumor cells. Toxicology data for humanized 4-1BB mice and cynomolgus monkeys showed that HuB6 could be tolerated up to a 180 mg/kg dose without systemic toxicity. CONCLUSIONS: This study demonstrated that HuB6 should be a suitable candidate for further clinical development and a potential agent for CRC immunotherapy.

Thyroid Disease Is Associated with Higher Age-Related Macular Degeneration Risk: Results from a Meta-Analysis of Epidemiologic Studies.

BACKGROUND: Although epidemiologic studies have suggested that thyroid disease may be a risk factor for age-related macular degeneration (AMD), this finding is still controversial. OBJECTIVES: The aim of this meta-analysis was to investigate whether an association exists between thyroid disease and medication and AMD in epidemiologic studies. METHODS: We searched PubMed, EMBASE, and Google Scholar from their inception to March 2020 for cross-sectional, case-control, and cohort studies that assessed thyroid function and AMD risk. Data from selected studies were extracted, and a meta-analysis was performed using fixed-effects or random-effects models. The statistical heterogeneity (I2) among studies and the possibility of publication bias were evaluated. If I2 >50%, a significant heterogeneity existed among studies, and a random-effects model was used to calculate the pooled RR. Otherwise, a fixed-effects model was performed. RESULTS: A total of 13 epidemiologic studies that consisted of 7 thyroid disease and 7 thyroid medication studies were included. Statistically significant heterogeneity was observed in the study results (I2thyroid disease = 80.1%; I2thyroid medication = 69.0%). A significant positive association was found between thyroid disease and AMD, with an overall relative risk (RR) of 1.25 (95% CI: 1.02, 1.54). However, there was no statistical association between thyroid medication and AMD risk (pooled RR 1.26 [95% CI: 0.92-1.72]). Egger's test indicated that there was no significant publication bias for thyroid disease (p = 0.889) or thyroid medication (p = 0.226). CONCLUSIONS: Our findings indicate that thyroid disease is associated with higher AMD risk. Thyroid disease prevention strategies may have a significant effect on the prevention of AMD and warrant further evaluation.

Upregulated ß-arrestin1 predicts poor prognosis and promotes metastasis via AKT/ERK signaling pathway in gastric cancer.

BACKGROUND: ß-Arrestins have been found to regulate cell proliferation, invasion and migration; transmit anti-apoptotic survival signals; and affect other characteristics of tumours. However, their role in gastric cancer (GC) is not clear. We investigated the role and mechanism of ß-arrestins in the regulation of GC. METHODS: We first examined ß-arrestins mRNA levels in 17 pairs of GC tissues by qRT-PCR. We also used immunohistochemistry to further examine the expression of ß-arrestins in 60 paraffin-embedded primary GC tissues and 20 normal gastric tissues. Then, the function of ß-arrestin1 was investigated in vitro and in vivo. RESULTS: ß-Arrestin1 was upregulated in GC tissue and was associated with tumour stage, lymph node metastasis, invasion depth and patient sex. High expression of ß-arrestin1 expression predicted poor prognosis in GC. ß-Arrestin1 promoted GC cell proliferation, migration and invasion, and it suppressed E-cadherin expression and upregulated Vimentin expression via AKT/ERK signalling pathway. The in vivo metastasis assays showed that knockdown of ß-arrestin1 reduced lung metastasis and inhibited EMT. CONCLUSION: The upregulation of ß-arrestin1 predicts poor prognosis and promotes metastasis and epithelial-mesenchymal transition in GC through AKT/ERK signalling pathway. This study may provide therapeutic advances for the treatment and early diagnosis of patients with metastatic GC.

The Wnt3a/ß-catenin/TCF7L2 signaling axis reduces the sensitivity of HER2-positive epithelial ovarian cancer to trastuzumab.

Epithelial ovarian cancer (EOC) is one of the most common and lethal gynecological cancers. Novel therapeutic agents have been developed for EOC, but patient survival remains poor. Trastuzumab has been approved for breast and gastric cancers with high expression of human epidermal growth factor receptor 2 (HER2), but it has not achieved any clinical success in EOC. Dysregulated Wnt/ß-catenin signaling is involved in cancer development, but whether it plays a role in EOC resistance to trastuzumab remains largely unknown. Here, we observed that high expression of Wnt3a, ß-catenin and TCF7L2, which can form a signaling axis in the Wnt/ß-catenin pathway, commonly existed in HER2-positive EOC tissue samples and was correlated with a poor patient prognosis. Cell proliferation and migration assays and nude mouse xenograft model experiments demonstrated that the Wnt3a/ß-catenin/TCF7L2 signaling axis promoted tumor cell growth and metastasis and reduced tumor sensitivity to trastuzumab. Analysis of downstream Akt signaling suggested that the function of the Wnt3a/ß-catenin/TCF7L2 signaling axis was mediated, at least in part, through increasing Akt phosphorylation. Overall, this study reveals a crucial role for the Wnt3a/ß-catenin/TCF7L2 signaling axis in EOC resistance to trastuzumab and the potential application of HER2-targeted drugs combined with inhibitors of this signaling axis for EOC treatment.

Intrinsically altered lung-resident γδT cells control lung melanoma by producing interleukin-17A in the elderly.

Cancer is an age-associated disease, potentially related to the altered immune system of elderly individuals. However, cancer has gradually decreased incidence in the eldest globally such as the most common lung cancer, the mechanisms of which remain to be elucidated. In this study, it was found that the number of lung-resident γδT cells was significantly increased with altered gene expression in aged mice (20-24 months) versus young mice (10-16 weeks). Aged lung Vγ4+ and Vγ6+ γδT cells predominantly produced interleukin-17A (IL-17A), resulting in increased levels in the serum and lungs. Moreover, the aged mice exhibited smaller tumors and reduced numbers of tumor foci in the lungs after challenge with intravenous injection of B16/F10 melanoma cells compared with the young mice. Aged lung Vγ4+ and Vγ6+ γδT cells were highly cytotoxic to B16/F10 melanoma cells with higher expression levels of CD103. The markedly longer survival of the challenged aged mice was dependent on γδT17 cells, since neutralization of IL-17A or depletion of indicated γδT cells significantly shortened the survival time. Consistently, supplementation of IL-17A significantly enhanced the survival time of young mice with lung melanoma. Furthermore, the anti-tumor activity of aged lung γδT17 cells was not affected by alterations in the load and composition of commensal microbiota, as demonstrated through co-housing of the aged and young mice. Intrinsically altered lung γδT17 cells underlying age-dependent changes control lung melanoma, which will help to better understand the lung cancer progression in the elderly and the potential use of γδT17 cells in anti-tumor immunotherapy.

HS-ß-cyclodextrin-functionalized Ag@Fe3O4@Ag nanoparticles as a surface-enhanced Raman spectroscopy substrate for the sensitive detection of butyl benzyl phthalate.

In recent years, there have been incidents involving the illegal addition of phthalic acid esters (PAEs) to liquors. It is well known that PAEs such as butyl benzyl phthalate (BBP) have estrogen-like effects, so high PAE levels in the body can lead to a decreased sperm count in males and altered sexual organ development in children, for example. The rapid detection of PAEs in liquor is therefore an important task. Compared with traditional methods of testing for PAEs, surface-enhanced Raman spectroscopy (SERS) offers higher sensitivity and the ability to search for chemical fingerprints, allowing the rapid detection of particular PAEs. In the present work, we synthesized Ag@Fe3O4@Ag/ß-cyclodextrin (CD) nanoparticles for use as a SERS-active substrate. Fe3O4 aggregates quickly under the influence of an external magnetic field, making it possible to magnetically separate out the NPs, which simplifies sample processing. The detection limit of the system for PAEs is also improved because the ß-CD acts as a functional group with a cavity structure that is capable of adsorbing BBP to form a host (ß-CD)-guest (BBP) complex. This substrate was shown to possess good repeatability and sensitivity when using malachite green (MG) as a probe molecule. Furthermore, the nanoparticle-based SERS substrate permitted the detection of BBP down to a level of 1.3 mg/kg in liquor, which is low enough to be able to detect BBP in real-world liquor samples. We expect that this method will prove useful for the rapid detection of PAEs in food. Graphical abstract.

Liquid-liquid interfacial self-assembled Au NP arrays for the rapid and sensitive detection of butyl benzyl phthalate (BBP) by surface-enhanced Raman spectroscopy.

Recent years have seen a large number of incidents involving the contamination of liquor with phthalate plasticizers (PAEs). There is therefore an urgent need to develop novel analytical strategies for the rapid and sensitive detection of PAEs. The PAE butyl benzyl phthalate (BBP) is very harmful to the human body, so we developed a surface-enhanced Raman spectroscopy (SERS) platform for the rapid detection of BBP in liquor based on liquid-liquid extraction and the simultaneous self-assembly of arrays of Au nanoparticles (Au NPs) at an organic/aqueous interface. The self-assembly of Au NPs occurs under the influence of the surface tension at the interface between the immiscible solvents. In the first step of the strategy, cyclohexane (CYH) is mixed with BBP-containing liquor to extract the BBP. Then the self-assembly of Au NPs at an organic/aqueous interface is induced using the CYH supernatant as the organic phase, a colloid of Au NPs as the aqueous phase, and ethanol as the inducer. During this process, the BBP molecules extracted from the liquor participate directly in the Au NP self-assembly process, which causes the analytes to be loaded into SERS-active nanogaps in the Au NP arrays, thus permitting the sensitive detection of BBP. BBP levels as low as 1.3 mg/kg in the liquor were detected using this method. Fifteen batches of the assembled SERS platform produced a relative standard deviation of 10.58% in the SERS intensity of the peak at 1178 cm-1 generated in the presence of 0.08 ppm crystal violet, indicating that this strategy possesses good reproducibility. Furthermore, interfacial assembly allowed the dual-analyte detection of BBP in the organic phase and an edible pigment (sunset yellow) in the aqueous phase to be achieved with high sensitivity and credible reproducibility using the SERS platform. Interfacial self-assembled SERS-active arrays therefore show great potential for the rapid and sensitive in situ detection of BBP in liquor samples. Graphical abstract ᅟ.

Clinicopathological and prognostic significance of circulating tumor cells in patients with lung cancer: a meta-analysis.

BACKGROUND: The prognostic significance of circulating tumor cells in patients with lung cancer is controversial. Therefore, we aimed to comprehensively and quantitatively assess the prognostic role of CTCs in patients with lung cancer. METHODS: The relevant literature was searched using PubMed, the Cochrane database and the China National Knowledge Internet database (up to June 2016). Using Review Manager 5.1.2, a meta-analysis was performed using hazard ratio (HR), odds ratio (OR) and 95% confidence interval (CI) as effect values. RESULTS: Thirty studies comprising 2,060 patients with lung cancer were analyzed. The pooled HR values showed that circulating tumor cells were significantly correlated with overall survival (HR =2.63, 95% CI [2.04, 3.39]) and progression-free survival (HR =3.74, 95% CI [2.49, 5.61]) in these patients. Further subgroup analyses were conducted and categorized by sampling time, detection method, and histological type; these analyses showed the same trend. The pooled OR values showed that circulating tumor cells were associated with non small cell lung cancer stage(OR = 2.11, 95% CI [1.42, 3.14]), small cell lung cancer stage (OR = 10.91, 95% CI [4.10, 29.06]), distant metastasis (OR =7.06, 95%CI [2.82, 17.66]), lymph node metastasis (OR =2.31, 95% CI [1.19,4.46]), and performance status(OR =0.42, 95%CI [0.22, 0.78]). CONCLUSION: The detection of circulating tumor cells in the peripheral blood of patients with lung cancer can be indicative of a poor prognosis.

Commensal microbiota maintains alveolar macrophages with a low level of CCL24 production to generate anti-metastatic tumor activity.

Microbiota maintains host tissue homeostasis and influences tissue-resident macrophages. However, the mechanisms by which commensal bacteria in regulating the alveolar macrophages remain unclear. Here, by using an antibiotic-treated (Abt) mouse model, we found commensal bacteria depletion induced lower frequencies and numbers of alveolar macrophages. This effect was accompanied by the altered levels of genes involved in several biological pathways, including M2 macrophage polarization, as determined by gene expression analysis. Alveolar macrophages from the Abt mice had higher protein and gene levels of Arg1, CCL24, IL-13, IL-10, IL-6 and IL-1ß, which could be recovered to normal levels by reconstructing commensal bacteria in the upper respiratory of Abt mice. Moreover, alveolar macrophages performed significant enhancement of M2 functions, especially CCL24 secretion, in the Abt mice challenged with B16/F10 melanoma. Adoptive transfer of normal alveolar macrophages or antibody neutralization of CCL24 significantly recovered the decrease of γδT17 cells and rescued the defect anti-tumor response of Abt mice, indicating the elevated amount of alveolar macrophage-derived CCL24 inhibited γδT cell mediated anti-tumor response. In conclusion, we demonstrated the ability of commensal bacteria to maintain alveolar macrophages with a low level of CCL24 production, which was necessary for the normal anti-tumor response in the lung.

Microbiota modulate tumoral immune surveillance in lung through a γδT17 immune cell-dependent mechanism.

Commensal bacteria are crucial to maintain immune homeostasis in mucosal tissues and disturbances in their ecology can affect disease susceptibility. Here, we report evidence that commensal bacteria shape the efficiency of immune surveillance in mucosal tissues. Antibiotic-treated (Abt) mice were more susceptible to development of engrafted B16/F10 melanoma and Lewis lung carcinoma, exhibiting a shortened mean survival time with more numerous and larger tumor foci in the lungs. The defective antitumor response of Abt mice was independent of dehydration caused by antibiotics. Host defenses relied upon intact commensal bacteria with no class specificity. Mechanistic investigations revealed a defective induction of the γδT17 cell response in lungs of Abt mice; here, more aggressive tumor development was observed, possibly related to a reduction in IL6 and IL23 expression there. Adding normal γδT cells or supplementing IL17 restored the impaired immune surveillance phenotype in Abt mice. Overall, our results demonstrated the importance of commensal bacteria in supporting the host immune response against cancer, defined an important role for γδT17 responses in the mechanism, and suggested deleterious effects of antibiotic treatment on cancer susceptibility and progression.

Elevated dopamine D2 receptor in prefrontal cortex of CUMS rats is associated with downregulated cAMP-independent signaling pathway.

Because depression is associated with significant morbidity and functional disability, it is important to reveal the mechanism of action. A variety of studies have suggested the involvement of dopaminergic receptors in the pathophysiological mechanism of non-stress-associated depression-like behavior in rodents. Nevertheless, controversy exists about whether chronic stress acts on dopaminergic receptors in the prefrontal cortex. Thus, we investigated the level of dopamine D2 receptors (DRD2) and the possible mechanisms involved in a chronic unpredictable mild stress (CUMS) rat model of depression. The results showed CUMS-induced, depression-like symptoms in the rat, characterized by reduced sucrose consumption and body mass, and increased duration of immobility in a forced swimming test. Moreover, chronic stress upregulated the expression of DRD2 but downregulated protein kinase A (PKA), transcription factor cAMP response element binding protein (CREB), and phospho-CREB (p-CREB) in the prefrontal cortex, as demonstrated by Western blot. Notably, in the rat model of depression, decreased cyclic adenine monophosphate (cAMP) levels and PKA activity were present at the same time, which is consistent with clinical findings in depressed patients. Our findings suggested that dopaminergic system dysfunction could play a central role in stress-related disorders such as depression.

Prognostic role of human epidermal growth factor receptor in gastric cancer: a systematic review and meta-analysis.

BACKGROUND AND AIMS: Human epidermal growth factor receptor (EGFR) and HER2 (ErbB2) both belong to EGFR family, which are overexpressed in a significant proportion of cases of gastric cancer (GC). Various studies have evaluated the prognostic value of EGFR or HER level in GC. However, the overall test performance remains unclear. We undertook this study to perform a systematic review and meta-analysis of prognostic cohort studies evaluating the use of EGFR or HER2 as a predictor of survival time in patients with GC. METHODS: Eligible studies were identified through multiple search strategies. Studies were assessed for quality using the Newcastle-Ottawa Tool. Data were collected comparing overall survival (OS) in patients with high and low EGFR or HER2 level. Studies were pooled and summary hazard ratios were calculated. RESULTS: Studies were listed twice if they provided overall survival data for both EGFR and HER2. Eight studies (seven for EGFR and eight for HER2) were included. Two distinct groups were pooled for analysis and revealed that high EGFR, HER2 levels predicted poor overall (HR = 1.66, 95% CI: 1.35-2.02) and (HR = 1.43, 95% CI: 1.09-1.88) survival. No publication bias was found. CONCLUSIONS: This meta-analysis result suggested that EGFR or HER2 should have significant predictive ability for estimating overall survival in GC patients and may be useful for defining prognosis of GC patients.

5,6,7,5'-Tetra-meth-oxy-3',4'-methyl-ene-dioxy-flavone monohydrate.

THE TITLE COMPOUND [SYSTEMATIC NAME: 5,6,7-trimeth-oxy-2-(7-meth-oxy-1,3-dihydro-2-benzofuran-5-yl)-4H-chromen-4-one monohydrate], C(20)H(18)O(8)·H(2)O, was isolated from the popular Chinese medicinal plant Entada phaseoloides. In the crystal, inversion-related mol-ecules are joined by pairs of weak C-H⋯O hydrogen bonds. The dimers are further inter-connected by a bridging water mol-ecule via weak C-H⋯O(water) and pairs of (O-H)(water)⋯O hydrogen bonds into a linear tape running parallel to the b axis.

[Effects of low-dose aspirin on primary prevention of cardiovascular events: a systematic review].

OBJECTIVE: To evaluate the effect and safety of low-dose aspirin for primary prevention of cardiovascular events. METHODS: We searched for randomized controlled trials (RCT) in the following electronic databases: MEDLINE, EMbase, the Cochrane Library (Issue 3, 2008), CBM, CNKI. Quality assessment and data extraction were conducted by two reviewers independently. All data were analyzed using Review Manager 4.2. RESULTS: Six studies (TPT, HOT, PPP, WHS, POPADAD, J-PAD) involving a total of 72,466 participants met the inclusion criteria. Meta-analysis results showed that: (1) Compared with placebo, the incidences of total cardiovascular events (RR = 0.85, 95% CI: 0.80-0.92), stroke (RR = 0.87, 95% CI: 0.77-0.98), nonfatal stroke (RR = 0.81, 95% CI: 0.70-0.95) and transient ischemic attack (RR = 0.76, 95% CI: 0.64-0.90) were significantly lower in low-dose aspirin group than those in placebo control group (all P < 0.05). (2) Nonfatal myocardial infarction (RR = 0.89, 95% CI: 0.77-1.02), death from cardiovascular causes (RR = 0.98, 95% CI: 0.86-1.13) and death from any cause (RR = 0.95, 95% CI: 0.88-1.02) were similar between the 2 groups (all P > 0.05). (3) The risk of coronary heart disease was reduced in low-dose aspirin group in the elderly (RR = 0.81, 95% CI: 0.70-0.94, P < 0.05). (4) The risk of bleeding was higher in low aspirin group compared to placebo group (RR = 1.15, 95% CI: 1.12-1.18, P < 0.01). CONCLUSIONS: Low-dose aspirin use could reduce the incidences of total cardiovascular events, stroke, nonfatal stroke and transient ischemic attack but increase the risk of bleeding, the incidence of nonfatal myocardial infarction, death from cardiovascular causes and death from any cause was not affected by low-dose aspirin use. Low-dose aspirin use was also significantly reduced the risk of coronary heart disease in the elderly.