5-min mindfulness audio induction alleviates psychological distress and sleep disorders in patients with COVID-19.

BACKGROUND: Mindfulness meditation is beneficial to mitigate the negative effects of the coronavirus disease 2019 (COVID-19) pandemic in the general population, but no study examined such meditation in the COVID-19 patients themselves. AIM: To explore the short-term efficacy of mindfulness meditation in alleviating psychological distress and sleep disorders in patients with COVID-19. METHODS: This prospective study enrolled patients with mild COVID-19 treated at Wuhan Fangcang Hospital in February 2020. The patients were voluntarily divided into either a mindfulness or a conventional intervention group. The patients were evaluated before/after the intervention using the Short Inventory of Mindfulness Capability (SMI-C), Hospital Anxiety and Depression Scale (HADS), and Pittsburgh Sleep Quality Index (PSQI). RESULTS: Seventy-five participants were enrolled in this study, with 43 and 32 in the mindfulness and conventional groups, respectively. Before the intervention, there were no differences in SMI-C, HADS, or PSQI scores between the two groups. After the 2-wk intervention, the mindfulness level (from 30.16 ± 5.58 to 35.23 ± 5.95, P < 0.001) and sleep quality (from 12.85 ± 3.06 to 9.44 ± 3.86, P < 0.001) were significantly increased in the mindfulness group. There were no differences in the conventional group. After the intervention, the mindfulness level (35.23 ± 5.95 vs 31.17 ± 6.50, P = 0.006) and sleep quality (9.44 ± 3.86 vs 11.87 ± 4.06, P = 0.011) were significantly higher in the mindfulness group than in the conventional group. Depression decreased in the mindfulness group (from 14.15 ± 3.21 to 12.50 ± 4.01, P = 0.038), but there was no difference between the two groups. CONCLUSION: Short-term mindfulness meditation can increase the mindfulness level, improve the sleep quality, and decrease the depression of patients with COVID-19.

Resilience as a Mediator of the Association Between Perceived Stigma and Quality of Life Among People With Inflammatory Bowel Disease.

Background: Improving Quality of Life (QOL) is an essential objective in the management of inflammatory bowel disease. An accumulating body of research has been conducted to explore the association between perceived stigma and QOL among patients with chronic illness. Still, underlying mechanisms behind this pathway have not been thoroughly examined. Objective: To investigate (a) the effect of perceived stigma on QOL among patients with inflammatory bowel disease; and (b) the mediating role of resilience in the association between perceived stigma and QOL. Methods: This cross-sectional study included a convenient sample of patients diagnosed with inflammatory bowel disease from four tertiary hospitals in Jiangsu Province, China. Patients completed the Perceived Stigma Scale in Inflammatory Bowel Disease (PSS-IBD), the Resilience Scale for Patients with Inflammatory Bowel Disease (RS-IBD), and the Inflammatory Bowel Disease Questionnaire (IBDQ). A bootstrapping analysis was implemented using the SPSS macro PROCESS. Results: A total of 311 patients with Cohn's disease and ulcerative colitis participated in this study, and 57.6% were men. The mean disease duration was 3.51 ± 1.04 years. Approximately 40% of the sample exceeded the criterion score for moderate stigma. Patients who perceived moderate or severe stigma reported lower QOL compared with those with mild stigma. After controlling for sociodemographic and clinical variables, we observed that perceived stigma was negatively associated with resilience. Moreover, resilience was found to mediate the relationship between perceived stigma and all aspects of QOL. Conclusions: These findings suggested that QOL of patients with inflammatory bowel disease was associated with perceived stigma and resilience and identified the mediating effects of resilience in the relationship between perceived stigma and QOL. Furthermore, this suggests that integrating intervention techniques to target resilience into the QOL improvement program of individuals with perceived stigma is possible.

Saturated fatty acids activate the inflammatory signalling pathway in Schwann cells: Implication in sciatic nerve injury.

Sciatic nerve injury affects quality of life. Many immune cells and inflammatory cytokines have been reported to be involved in sciatic nerve injury, but little is known about the ligands and receptors that trigger inflammatory responses. By using a modified sciatic nerve clamp injury method, we found that the recruitment of Schwann cells and the inflammatory response were enhanced after sciatic nerve injury. Toll-like receptor 4 (TLR4), one of the major members of the TLR family, is highly expressed in Schwann cells. Under certain conditions, myeloid differentiation protein 2 (MD2) binds to TLR4 on the membrane and plays important roles in the inflammatory response. The reductions in the recruitment of Schwann cells and the inflammatory response induced by the blockade of TLR4 or MD2 suggest that TLR4 and MD2 are involved in sciatic nerve injury. What are the endogenous signals that activate the inflammatory response? A large number of free saturated fatty acids (SFAs) are released from Schwann cells, adipocytes and the blood after sciatic nerve injury. Liang et al reported that Schwann cells can be stimulated by palmitic acid (PA). Here, we found that the expression and secretion of TNF-α and IL-6 were enhanced by PA treatment. Moreover, PA activated TLR4 signalling pathway-related proteins and stimulated a strong association between TLR4 and MD2. Blocking TLR4 or MD2 reversed the PA-induced inflammatory response and TLR4 downstream signalling pathway. Thus, we speculated that SFAs act as endogenous ligands that activate TLR4/MD2, thus triggering Schwann cell inflammation during sciatic nerve injury.

NLRP3 inflammasome is involved in nerve recovery after sciatic nerve injury.

The activation of the inflammasome plays an important role in the central nervous system. However, only a few studies have investigated the effects of inflammasome activation in the peripheral nerve, especially in the sciatic nerve, and the mechanism of this activation remains elusive. Moreover, how interleukin-1 beta (IL-1ß) is produced after sciatic nerve injury is also unknown. In our study, we aimed to investigate whether the nucleotide-binding oligomerization domain-like pyrin domain containing protein 3 (NLRP3) inflammasome is activated after sciatic nerve injury and to explore its role in sciatic nerve injury. The results of immunoblotting and immunofluorescence microscopy indicate that the NLRP3 inflammasome was activated after sciatic nerve injury in wild-type (WT) mice, as demonstrated by upregulated inflammasome-related components, e.g., NLRP3, procaspase-1 and ASC. Furthermore, upregulated inflammasome-related components cis-cleavage precursor IL-1ß (proIL-1ß) and precursor interleukin-18 (proIL-18) to IL-1ß and IL-18, contributing to the inflammatory response. Consequently, the inflammatory response after sciatic nerve injury in NLRP3 knockout (NLRP3-KO) mice was less severe than that in WT mice. Moreover, NLRP3-KO mice exhibited an increased sciatic functional index (SFI), which was determined by footprint analysis, suggesting that NLRP3 deficiency is beneficial to sciatic nerve recovery after injury. Therefore, our results indicate that NLRP3 is involved in the recovery from sciatic nerve injury and mediates the production of inflammatory factors, such as IL-1ß, after sciatic nerve injury.

Withdrawal Notice: Cryptotanshinone Suppresses Liver Fibrosis by Attenuating Epithelial-Mesenchymal Transition through Targeting Hedgehog Pathway

The article has been withdrawn at the request of the authors because some of the authors do not agree with the authorship. Bentham Science apologizes to the readers of the journal for any inconvenience this may have caused. The Bentham Editorial Policy on Article Withdrawal can be found at https://benthamscience.com/editorial-policies-main.php Bentham Science Disclaimer: It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submitting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript, the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Cross-regulation by TLR4 and T cell Ig mucin-3 determines severity of liver injury in a CCl4-induced mouse model.

Acute liver injury is a common pathological basis for a variety of acute liver diseases in the clinic, which can eventually lead to liver fibrosis and even liver failure. In this study, we found that T cell Ig and mucin domain protein 3 (Tim-3) and TLR4 receptors play important roles in CCl4-induced acute liver injury. Tim-3 is a negative regulator that is expressed by T cells and macrophages. Using antibodies against Tim-3 (anti-Tim-3 Ab), we studied the Tim-3 signal in an animal model of acute liver injury and found that a large number of inflammatory factors were upregulated. In vitro experimental data shown that anti-Tim-3 Ab treatment increased interferon-É£ production by concanavalin A (ConA)-stimulated spleen T cells, and we found that the expression level of interleukin (IL)-6 was increased in a macrophage/spleen T cell coculture system, while administration of galectin-9 (Gal-9, a Tim-3 ligand) reduced the IL-6 production. This indicates the importance of the Tim-3/Gal-9 signalling pathway in maintaining hepatic homeostasis. The Tim-3 signalling pathway inhibits TLR4-mediated NF-κB activity, and an anti-Tim-3 Ab does not affect the liver injury in TLR4-deficient mice. Regulation between Tim-3 and TLR4 determines the severity of liver damage. The negative regulation of Tim-3 reflects the protective mechanisms of patients with impaired liver function, and these results provide important information about innate and adaptive responses in the regulation of liver damage. This finding is potentially important for the study of early liver injury.

Imiquimod enhances DNFB mediated contact hypersensitivity in mice.

Imiquimod (Imiq) is a synthetic imizoquinoline compound which can act on Toll-like receptor (TLR)7 and transduce signals involved in cell activation. We investigated the role of Imiq on contact hypersensitivity (CHS) and explored the potential mechanisms of mast cells involved in the process. Topical application of Imiq cream augmented DNFB mediated CHS in C57BL/6 mice. Imiq application induced skin inflammation and increased the number of dendritic cells (DCs) in the draining lymph nodes (DLNs). The splenic cell proliferation to DNBS in DNFB and Imiq treated mice was greater than that in mice of DNFB treatment alone. Peritoneal cell-derived mast cells (PCMCs) expressed TLR7 mRNA. The results from toluidine blue staining for mast cells and histamine detection indicated that Imiq alone did not induce mast cell degranulation while Imiq plus DNFB significantly induced mast cell degranulation. Cromolyn, pyrilamine and cimetidine attenuated CHS reaction induced by Imiq. Our findings suggest that Imiq could augment the intensity of CHS reaction. The mechanisms underlying the effect may relate to histamine release by mast cells and induction of DC homing to DLNs. Blocking histamine action in early time of allergen contact is beneficial to the alleviation of CHS.

Imiquimod enhances the potency of an exogenous BM-DC based vaccine against mouse melanoma.

Dendritic cell (DC) vaccine is a potent immunotherapeutic approach for cancer treatment, but the clinical efficacy needs to be improved. In this study, we evaluated the combinational effect of Toll-like receptor 7 (TLR7) agonist Imiquimod and BM-DC vaccine against mouse melanoma and explored the potential mechanisms. We found that topical application of Imiquimod cream caused skin inflammation and enhanced exogenous BM-DC homing to draining lymph nodes. Imiquimod treatment enhanced DC vaccine efficacy against B16-OVA melanoma. The combinational modality enhanced cytotoxicity of splenic lymphocyte to tumor cells and inhibited CD4+FOXP3+Treg cell production. TLR7 mRNA expression was confirmed in both MC/9 mast cells and DCs. MC/9 cells treated by R837 (soluble form of Imiquimod) enhanced CD80, CD86, MHC-II and CCR7 expression on DCs. R837 inhibited B16-OVA cell growth in vitro. Our findings suggest that Imiquimod can be used as a potent adjuvant in the formulation of a DC-based tumor fighting vaccine. The mechanisms underlying these effects of Imiquimod are related with enhanced DC homing to DLNs, inhibition of Treg's production, direct tumor cell toxicity and synergistic function with mast cell in enhancing DC activation.

[Imiquimod combined with dendritic cell vaccine decreases Treg proportion and enhances anti-tumor responses in mice bearing melanoma].

Objective: To investigate the therapeutic effect of Toll-like receptor 7( TLR7) agonist imiquimod combined with dendritic cell( DC)-based tumor vaccine on melanoma in mice and the potential mechanism. Methods: Melanoma-bearing mouse models were established by subcutanous injection of B16-OVA cells into C57 BL /6 mice. DCs were isolated from mouse bone marrow and propagated in culture medium with recombinant mouse granulocyte-macrophage colony-stimulating factor( rm GM-CSF) and recombinant mouse interleukin-4( rm IL-4). DC vaccine( OVA-DC) was prepared by overnight incubation of DCs added with chicken ovalbumin. C57 BL /6 mice were separated into four groups which were treated with PBS,topical imiquimod application,OVA-DC intradermal injection and imiquimod plus OVA-DC,respectively. The tumor size was calculated by digital vernier caliper. Peripheral blood CD4~+FOXP3~+Tregs of the tumor-bearing mice was detected by flow cytometry. The cytotoxicity of splenic lymphocyte against B16-OVA was assessed in vitro by CCK-8 assay. Results: Compared with the other three groups,B16-OVA-bearing mice treated with imiquimod plus DC vaccine had the smallest tumor volume. The percentage of CD4~+FOXP3~+Tregs decreased significantly in the combined treated mice. The combined treatment enhanced significantly cytotoxicity of splenic lymphocytes against B16-OVA cells. Conclusion: Imiquimod combined with antigen-pulsed-DC vaccine could reduce CD4~+FOXP3~+Treg proportion and promote anti-tumor effect in mice with melanoma

[Establishment of a syngeneic mouse model of liver tumor stably expressing hepatitis B virus antigens].

OBJECTIVE: To establish a syngeneic mouse model of liver tumor stably expressing hepatitis B virus (HBV) antigens. METHODS: Melanoma cell line B16 cells were transfected with pLXSN-2HBV. Cells (named B16/HBV) stably and persistently expressing HBV surface (HBsAg) and core (HBcAg) antigens were identified. The cells were injected into the hepatic subcapsular space of fifteen C57BL/6J mice. The mice were divided into 3 groups, receiving 100, 1000 or 5000 cells in a total volume of 5 µl per mouse, respectively, five mice in each group. Two weeks after the tumor cell inoculation, serum samples from the mice were collected weekly and the serum concentration of HBsAg and anti-HBs was quantified by ELISA. The tumor growth in the mouse liver was monitored by a high-resolution ultrasound system. Expression of HBsAg and HBcAg in the tumor tissues was determined by immunohistochemistry. RESULTS: Liver tumors were formed in all the mice receiving 1000 and 5000 B16/HBV cells per mouse, and in 80% of the mice receiving 100 B16/HBV cells. HBsAg and anti-HBs were detectable in their sera from 2 weeks after tumor cell inoculation. The mice receiving 100 cells per mouse began to die 4 weeks, those receiving 1000 cells per mouse began to die 3 - 4 weeks and those receiving 5000 cells began to die 2 - 3 weeks after the cell inoculation. All the tumor cells expressed HBsAg and HBcAg. CONCLUSIONS: The B16/HBV cells stably and persistently express HBV antigens both in vitro and in vivo. A mouse model of transplanted liver tumor stably expressing HBV antigens has been successfully established by inoculation of those cells into the hepatic subcapsular space.

[Effect of tetramethylpyrazine on JAK-STAT signal transduction in cardiomyocyte hypertrophy].

AIM: To study the effect of Tetramethylpyrazine (TMZ), the extracts of Chinese Herbs which accelerat blood circulation, on JAK-STAT signal transduction in cardiomyocyte hypertrophy. METHODS: Cardiomyocyte hypertrophy Wistar rats model was established. The cardiomyocytes were seperated from one-day-old neonatal rats. The total RNA of cardiomyocytes was extracted by TRIzol reagent. Then the ANP mRNA were detected by RT-PCR. pJAK2, pJAK1 and pSTAT3 molecules were analysed by Western blot respectively. RESULTS: Statistics analysis showed that TMZ significantly decreased the expression of ANP mRNA in cardiomyocytes (P<0.01), and that TMZ also decreased the levels of pJAK2, pJAK1 or pSTAT3 (P<0.01). CONCLUSION: TMZ has the inhibitory effect on JAK-STAT signal transduction in cardiomyocyte hyertrophy, suggesting the clinical application of traditional Chinese medicine on cardiomyocyte hypertrophy treatment.

[Mast cell degranulation enhances exogenous dendritic cells homing to draining lymph nodes].

AIM: To establish a method of enhancing exogenous bone marrow derived dendritic cells (BM-DC) homing to draining lymph nodes by induction of local mast cell degranulation. METHODS: Compound 48/80 (c48/80) was injected into C57BL/6 scapular skin to induce local mast cell degranulation. BM-DC generated from bone marrow of syngenic mice were labeled with YG-Microspheres and injected into c48/80 or normal saline treated scapular skin. Cells derived from draining lymph nodes (DLN) were pooled together 48 h after BM-DC injection and stained with conjugated anti-CD11c. The efficiency of BM-DC homing to lymph nodes was analyzed by FCM. RESULTS: Mast cell degranulation was locally boosted by c48/80 injection and caused enhancement of the homing of exogenous BM-DC to DLN, with an increase of 67%+/-43%. The total cells in lymph nodes also increased significantly. The fold of increase was 55%+/-43%. CONCLUSION: Exogenous BM-DC homing to DLN can be boosted by inducing local skin mast cell degranulation.

TLR8 agonists stimulate newly recruited monocyte-derived cells into potent APCs that enhance HBsAg immunogenicity.

We previously reported that synthetic or natural Toll-like receptor (TLR) 7/8 agonists present within dead cells enhanced cell-associated antigen presentation both in vitro and in vivo. Here, we investigated the immunopotency of different chemically synthesized TLR7/8 agonists, Resiquimod, Gardiquimod, CL075, and CL097, on HBsAg immunogenicity. These agonists stimulated inflammatory monocyte-derived cells to become potent antigen-presenting dendritic cells (DCs), which augmented HBsAg specific T cell proliferation after they were conditioned with HBsAg. The TLR8 agonist CL075 and the TLR7/8 dual agonist CL097 showed more potent effects than the TLR7 agonist. Compared with alum adjuvant, when HBsAg mixed with CL075 was injected intramuscularly into mice, more monocyte-derived DCs carried antigens into draining lymph nodes and spleens. Specific Abs, particularly IgG2a, were significantly increased, and more IL-5 and IFN-gamma were produced by splenocytes and intrahepatic immunocytes in mice that received HBsAg mixed with CL075 and CL097. These results suggest that TLR8 agonists are good candidates to enhance recombinant HBsAg immunogenicity to induce specific humoral and cellular immune responses.

Exogenous dendritic cell homing to draining lymph nodes can be boosted by mast cell degranulation.

Dendritic cells (DCs), as potent antigen presenting cells, are increasingly used for immunotherapeutic approaches, predominantly in oncology. Low efficiency of injected Ag-pulsed DC homing to draining lymph nodes (DLNs) is one of the factors that affect the efficacy of therapy. As Langerhans cell emigration was enhanced after skin mast cell degranulation, we investigated the effect of local mast cell activation on exogenous bone marrow-derived DCs (BM-DCs) homing to DLNs. Product of activated MC/9 mast cells enhanced chemotaxis of BM-DCs to CCL21 in vitro. Intradermal injection of compound 48/80 (c48/80) induced local skin mast cell obvious degranulation and boosted exogenous BM-DC homing to DLNs. Both Ag-specific lymphocyte proliferation and TH1/TH2 cytokine production increased after HBsAg-pulsed BM-DC was injected into c48/80 pretreated mice. These results suggest that transferred DC homing to DLNs promoted by local mast cell degranulation may have potential application to improve DC-based immunotherapy.

Activity after site-directed mutagenesis of CD59 on complement-mediated cytolysis.

CD59 may inhibit the cytolytic activity of complement by binding to C8/C9 and protect host cell membranes against homologous membrane attack complex (MAC). However, CD59 is widely overexpressed on tumor cells, which has been implicated in tumorigenesis. The active site of CD59 relative to MAC is still confused. As reported the MAC binding site is located in the vicinity of a hydrophobic groove on the membrane distal face of the protein centered around residue W40. Here two site-directed mutagenesis were performed by overlapping extension PCR to delete residue W40 site (Mutant 1, M1) or to change C39W40K41 to W39W40W41 (Mutant 2, M2). Then we constructed mutant CD59 eukaryotic expression system and investigated their biological function on CHO cells compared with wild-type CD59. Stable populations of CHO cells expressing recombinant proteins were screened by immunotechnique. After 30 passages culturing, proteins could be tested. Dye release assays suggest that M1CD59 loses the activity against complement, while M2CD59 increases the anti-complement activity slightly. Results indicate that W40 of human CD59 is important to its activity, and prohibition of this site may be a potential way to increase complement activity and to treat tumors.