Portal vein tumour thrombosis radiotherapy improves the treatment outcomes of immunotherapy plus bevacizumab in hepatocellular carcinoma: a multicentre real-world analysis with propensity score matching.

Background: This study aimed to evaluate the efficacy and safety of sequential immune checkpoint inhibitors (ICIs) plus bevacizumab therapy after radiotherapy for portal vein tumour thrombosis (PVTT) in patients with hepatocellular carcinoma (HCC). Methods: Retrospective data were collected from 113 patients with HCC with PVTT. Patients in the PVTT radiotherapy (radiotherapy + ICIs + bevacizumab) and control groups (ICIs + bevacizumab) were enrolled according to propensity score matching (PSM) analysis (1:1). The differences in progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and potential factors affecting PFS between the groups were analysed. The adverse events (AEs) were compared between the two groups. Results: There were 47 patients in the two groups after PSM (1:1). The differences in neutrophil and lymphocyte counts, neutrophil-to-lymphocyte ratio (NLR), CRP, and CD4, CD8, and CD4-to-CD8 ratio before and after radiotherapy for PVTT (P < 0.05) in the PVTT radiotherapy group were significant. The patients in the PVTT radiotherapy group had a longer PFS (median, 9.6 vs. 5.4 months, P < 0.001), and the PFS rates of 3, 6, 9, and 12 months were 97.87% vs. 94.19%, 80.85% vs. 44.68%, 53.19% vs. 6.38%, and 23.40% vs. 0.00%, respectively (P < 0.001). There were also significant differences in the ORR (48.94% vs. 27.66%, P = 0.0339) and DCR (97.87% vs. 82.98%, P = 0.0141) between the two groups, and no serious AEs were observed. Multivariate Cox analysis showed that AFP expression, gross classification of HCC, PVTT type, extrahepatic metastasis, PVTT radiotherapy, and reduction in PVTT were independent factors influencing PFS (P < 0.05). Conclusions: Sequential ICIs plus bevacizumab therapy after radiotherapy for PVTT in patients with HCC is safe and feasible and may further prolong the PFS of patients.

Isolation and characterization of dihydropyran-ring containing meroterpenoids from Ganoderma lucidum and their inhibitory activity against renal fibrosis-related protein expression.

The Ganoderma lucidum mushroom, which has been used as a traditional medicine in China for more than 2000 years, is a source of many interesting natural product. In this study, the five undescribed minor meroterpenoids baoslingzhines F-J (1-5), containing a dihydropyran moiety, were isolated as racemic mixtures from the fruiting bodies of G. lucidum. These substances were structurally and stereochemically characterized by using spectroscopic and computational methods. Chiral HPLC was employed to separate the (+)- and (-)-antipodes. A survey of the activities against kidney fibrosis showed that both enantiomers of baoslingzhines F-J inhibit expression of renal fibrosis-related proteins, including fibronectin, collagen I and ɑ-SMA in TGF-ß1-induced rat kidney proximal tubular cells.

COX-2 and iNOS inhibitory epimeric meroterpenoids from Ganoderma cochlear and structure revision of cochlearol Q.

Four novel epimeric meroterpenoids, ganadone A (1), 3',10'-di-epi-ganadone A (2), 10'-epi-ganadone A (3), and 3'-epi-ganadone A (4) as well as another pairs of epimers, ganadone B (5) and 10'-epi-ganadone B (6), with a same basic skeleton compound ganadone C (7), together with two lactonized meroterpenoids, ganadones D and E (8 and 9) were isolated from the fruiting bodies of Ganoderma cochlear. Compounds 1-7 were constructed with fascinating adjacent 6',7'-bifuran ring system. Fortunately, we have revised our previously reported structure cochlearol Q, which was proposed pyrano[6',7'-b]pyran ring system into 6',7'-bifuran motif. All the isolates were characterized by analysis of HRESIMS, NMR spectroscopy and 1 was supported by X-ray crystallography analysis. The absolute stereochemistry of 1-9 were assigned by quantum chemical calculations. Biological evaluation of 1-9 showed that 5, 6, and 9 have significant anti-inflammatory potentials.

Meroterpenoids with a large conjugated system from Ganoderma lucidum and their inhibitory activities against renal fibrosis.

Baoslingzhines A-E (1-5), five new meroterpenoids were isolated from the fruiting bodies of Ganoderma lucidum. The structures including their absolute configurations were characterized by using spectroscopic and computational methods. Compound 1 is a novel trinormeroterpenoid featuring the presence of an unusual dihydronaphthalene representing an unprecedented meroterpenoid skeleton. Compounds 2-4 are mononormeroterpenoids characteristic of a large conjugated system. Among them, racemic 3 and 4 were separated by HPLC on chiral phase. Biological evaluation toward kidney fibrosis found that compounds 2 and (+)-3 could inhibit the expression of fibronectin and collagen I dose dependently in TGF-ß1-induced rat kidney proximal tubular cells (NRK-52e). Additionally, (+)-3 could also down regulate ɑ-SMA in a concentration dependent manner. Further investigation showed that 2 could inhibit Smad2 phosphorylation.

Spiroganodermaines A-G from Ganoderma species and their activities against insulin resistance and renal fibrosis.

Ganoderma mushrooms are a renowned Chinese medicine and functional food used worldwide. Seven undescribed spiro Ganoderma meroterpenoids spiroganodermaines A-G were isolated from Ganoderma species. Their structures were characterized by using spectroscopic, computational and X-ray diffraction methods. Biological studies showed that (+)-spiroganodermaine G significantly activates glucose uptake and IRS1 phosphorylation in insulin resistance C2C12 cells. Furthermore, (-)-spiroganodermaine G inhibits the expressions of fibronectin and α-SMA in TGF-ß1 induced NRK-52E cells. These findings demonstrate the potential of Ganoderma meroterpenoids as medicines and dietary supplements.

TNFα mediates stress-induced depression by upregulating indoleamine 2,3-dioxygenase in a mouse model of unpredictable chronic mild stress.

Depression is often preceded by exposure to stressful life events. Chronic stress causes perturbations in the immune system, and up-regulates production of proinflammatory cytokines, which has been proposed to be associated with the pathogenesis of clinical depression. However, the potential mechanisms by which stress-induced proinflammatory cytokines lead to the development of depression are not well understood. Here, we sought to screen the main proinflammatory cytokines and the potential mechanisms linking inflammation to depression-like behavior during unpredictable, chronic, mild stress (UCMS), in vivo. Mice were allocated into four groups in each separate experiment: saline-control, saline-UCMS, drug-control and drug-UCMS. Development of depression-like behavior was reflected as a reduction in sucrose preference, and increased immobility in both the forced swim and tail suspension tests. The following drugs were administered intraperitoneally: the pan-anti-inflammatory tetracycline derivative, minocycline (30 mg/kg, daily), the tumor necrosis factor (TNF)α monoclonal antibody, infliximab (10 mg/kg, twice weekly), and the indoleamine 2, 3-dioxygenase (IDO) inhibitor, 1-methyltryptophan (1-MT, 10 mg/mouse, daily). Plasma TNFα, IL-1ß and IL-18 increased significantly after the four-week UCMS exposure. Pretreatment of mice with minocycline completely blocked any upregulation. Concurrent with development of depression-like behaviors, the concentration of TNFα in plasma and the cerebral cortex increased remarkably. The tryptophan-degrading enzyme IDO was up-regulated in the cortex following UCMS exposure. Treatment of mice with minocycline, infliximab or 1-MT prevented the development of depression-like behaviors. Furthermore, blockade of TNFα inhibited expression of IDO and protected cortical neurons from UCMS-induced damage. These results suggest that TNFα plays a critical role in mediating UCMS-induced depression through up-regulation of IDO and subsequent damage of cortical neurons.

Involvement of inflammasome activation in lipopolysaccharide-induced mice depressive-like behaviors.

AIMS: The NLRP3 inflammasome is a cytoplasmic multiprotein complex of the innate immune system that regulates the cleavage of interleukin-1ß and interleukin-18 precursors. It can detect a wide range of danger signals and trigger a series of immune-inflammatory reactions. There were plenty of studies indicating that activation of the immune system played pivotal roles in depression. However, the underlying mechanisms of immune-depression interactions remained elusive and there was no report about the involvement of inflammasome activation in depression. METHODS: We established an acute depression mouse model with lipopolysaccharide to explore the involvement of inflammasome activation in depression. RESULTS: The lipopolysaccharide-treated mice displayed depressive-like behaviors and pro-inflammatory cytokine interleukin-1ß protein and mRNA levels significantly increased. The NLRP3 inflammasome mRNA expression level also significantly elevated in depressed mice brain. Pretreatment with the NLRP3 inflammasome inhibitor Ac-YVAD-CMK significantly abrogated the depressive-like behaviors induced by lipopolysaccharide. CONCLUSION: These data suggest for the first time that the NLRP3 inflammasome is involved in lipopolysaccharide-induced mice depressive-like behaviors. The NLRP3 inflammasome may be a central mediator between immune activation and depression, which raises the possibility that it may be a more specific target for the depression treatments in the near future.

[Roles of cytokines in stress-induced depression].

Depression is a very common mental health problem in our modern society. Stress is involved in the provocation of depression. The pathogenesis of depressive disorder is still not well known. The development of neuroendocrine immunology opens a new sight for clarification of mechanism underlying stress-induced depression. Chronic stress activates peripheral and central immune systems accompanied with the release of inflammatory mediators, including cytokines. Activated immune system mediates the process of depression through the interaction with neuron system and neuroendocrine system, including regulating the monoamine neurotransmitter system in synthesis, metabolism and reuptake, inducing the overactivation of hypothalamus-pituitary-adrenal (HPA) axis and its negative feedback regulation, and reducing neurogenesis. This present paper reviews the cytokines mechanisms underlying stress-induced depression.

Dexamethasone induces rapid promotion of norepinephrine­mediated vascular smooth muscle cell contraction.

The aim of the present study was to identify the rapid effect of dexamethasone (Dex) on norepinephrine (NE)­mediated contraction of vascular smooth muscle cells (VSMCs) and to establish the underlying mechanism(s). Rat VSMCs were preincubated with lipopolysaccharide to simulate acute septic shock. Myosin light chain (MLC20) phosphorylation of VSMCs was detected by western blot analysis to observe the effects of Dex on NE­mediated contraction. Activation of the RhoA/ RhoA kinase (ROCK), extracellular signal­regulated kinase (ERK) and p38 signaling pathways was detected by western blot analysis to explore the mechanism. It was identified that Dex rapidly promoted NE­induced phosphorylation of MLC20 in VSMCs and this effect may be non­genomic. The RhoA/ROCK, ERK and p38 pathways were demonstrated to be important for the rapid effect of Dex­induced promotion of NE­mediated contraction in VSMCs. The present results indicate that Dex may rapidly reverse the hyporeactivity of vasoconstriction to NE in vitro and this effect may be mediated by specific non­genomic mechanisms through increased activation of the RhoA/ROCK, ERK and p38 signaling pathways.

Inducible nitric oxide synthase is involved in the modulation of depressive behaviors induced by unpredictable chronic mild stress.

BACKGROUND: Experiences and inflammatory mediators are fundamental in the provocation of major depressive disorders (MDDs). We investigated the roles and mechanisms of inducible nitric oxide synthase (iNOS) in stress-induced depression. METHODS: We used a depressive-like state mouse model induced by unpredictable chronic mild stress (UCMS). Depressive-like behaviors were evaluated after 4 weeks of UCMS, in the presence and absence of the iNOS inhibitor N-(3-(aminomethyl)benzyl)acetamidine (1400 W) compared with the control group. Immunohistochemistry was used to check the loss of Nissl bodies in cerebral cortex neurons. The levels of iNOS mRNA expression in the cortex and nitrites in the plasma were measured with real-time reverse transcription PCR (RT-PCR) and Griess reagent respectively. RESULTS: Results showed that the 4-week UCMS significantly induced depressive-like behaviors, including decreased sucrose preference in a sucrose preference test, increased duration of immobility in a forced swim test, and decreased hole-searching time in a locomotor activity test. Meanwhile, in the locomotor activity test, UCMS had no effect on normal locomotor activities, such as resting time, active time and total travel distance. Furthermore, the levels of iNOS mRNA expression in the cortex and nitrites in the plasma of UCMS-exposed mice were significantly increased compared with that of the control group. Neurons of cerebral cortex in UCMS-exposed mice were shrunken with dark staining, together with loss of Nissl bodies. The above-mentioned stress-related depressive-like behaviors, increase of iNOS mRNA expression in the cortex and nitrites in the plasma, and neuron damage, could be abrogated remarkably by pretreating the mice with an iNOS inhibitor (1400 W). Moreover, neurons with abundant Nissl bodies were significantly increased in the 1400 W + UCMS group. CONCLUSIONS: These results support the notion that stress-related NO (derived from iNOS) may contribute to depressive-like behaviors in a mouse model, potentially concurrent with neurodegenerative effects within the cerebral cortex.

The glycan profile of endothelial cells in the present of tumor-conditioned medium and potential roles of beta-1,6-GlcNAc branching on HUVEC conformation.

Endothelium plays a vital role in the logistics of the immune system, as well as the maintenance of the homeostasis. The major objective of this study is to unravel the relationship between expression changes of carbohydrate structures and the dysfunction of human umbilical vein endothelial cells (HUVEC) stimulated with tumor-conditioned medium (TCM), which is involved in tumor cell extravasation. Using flow cytometry (FCM) assay, the expression profiles of a selected group of 9 carbohydrate structures have been determined in HUVEC under control conditions and TCM-treated conditions, six of which increased significantly in expression after induction. Particularly, the expression level of beta-1,6-GlcNAc branching glycan was extremely higher after the stimulation. In parallel, the conformation change of HUVEC monolayer has been detected with inverted phase contrast microscopy and confocal microscopy. Under TCM stimulation, the actin cytoskeleton underwent rearrangement and formed abundant stress fiber within cells; therefore cell contraction was induced, which resulted in paracellular gap formation and barrier dysfunction. We furthered our study to investigate the mechanism underlying the conformation change of HUVEC. The results demonstrated that TCM induced the increase in beta-1,6-GlcNAc branching expression of PECAM-1, accompanied by the tyrosine phosphorylation of PECAM-1. The downstream effector RhoA was activated in consequence of the activation of PECAM-1. In conclusion, our results strongly suggested that the carbohydrate composition of endothelial cell surface is very important for the cells to exert their physiological effects correlated with cancer extravasation.

Expression of polymeric immunoglobulin receptor mRNA and protein in human paneth cells: Paneth cells participate in acquired immunity.

OBJECTIVE: Paneth cells are important effectors of intestinal innate immunity. It has been generally accepted that Paneth cells do not participate in the synthesis of polymeric immunoglobulin receptor (pIgR) or the secretion of immunoglobulin A (IgA) in the small intestine. However, we have previously shown that pIgR is specifically localized in Paneth cells of the rat small intestine. We therefore investigated the possibility that pIgR is also localized in human Paneth cells. METHODS: Double-labeled fluorescent immunohistochemistry and double-labeled fluorescent in situ hybridization were used to determine RNA and protein expression in normal human small intestine. RESULTS: Both pIgR mRNA and protein were colocalized with lysozyme in normal human Paneth cells. Furthermore, IgA was colocalized with lysozyme in the secretory granules of human Paneth cells. CONCLUSIONS: This is the first study to demonstrate that pIgR and IgA are colocalized in the secretory granules of human Paneth cells. These findings suggest that, in addition to their well-recognized role in innate immunity, Paneth cells are involved in IgA-mediated acquired immunity in the gastrointestinal tract. Furthermore, these results add to accumulating evidence that Paneth cells participate in intestinal inflammation.