Triptolide Inhibits Expression of Inflammatory Cytokines and Proliferation of Fibroblast-like Synoviocytes Induced by IL-6/sIL-6R-Mediated JAK2/STAT3 Signaling Pathway

Triptolide,a component of the Chinese herb Tripterygium wilfordii Hook F,has been proved to be effective in the treatment of rheumatoid arthritis(RA).However,its underlying mechanisms on RA have not yet been well established.We observed the inhibitory effect of triptolide on the expression of inflammatory cytokines and proliferation of fibroblast-like synoviocytes(FLS)induced by the complex of interleukin-6(IL-6)and the soluble form of the IL-6 receptor(sIL-6R).Furthermore,to clarify the underlying mechanisms,we treated FLS with the Janus-activated kinase 2(JAK2)inhibitor/signal transducer and activator of transcription 3(STAT3)activation blocker AZD1480.In this study,immunohistochemical staining was used to identify vimentin(+)and CD68(−)in FLS.The FLS proliferation was measured by cell proliferation assay,and the cell cycles were analyzed by flow cytometry.Furthermore,ELISA was used to detect the expression of the inflammatory factors in culture solution.The expression levels of p-JAK2,JAK2,p-STAT3 and STAT3 were investigated through Western blotting analysis.The results showed that IL-6/sIL-6R significantly increased the cell proliferation and expression of inflammatory cytokines,including IL-6,interleukin-1β(IL-1β)and vascular endothelial growth factor(VEGF).Triptolide or AZD1480 inhibited the cell proliferation and inflammatory cytokine expression in IL-6/sIL-6R-stimulated FLS by suppressing JAK2/STAT3.The study suggested that the physiological effects of triptolide on RA were due to its contribution to the inhibition of the inflammatory cytokine expression and FLS proliferation by suppressing the JAK2/STAT3 signaling pathway.It may provide an innovative insight into the effect of triptolide in preventing RA pathogenesis.

Molecular mechanism in the formation of pre-metastatic niches

Pre-metastatic niches(PMNs)are the microenvi-ronments prepared by the interaction of tumor cells and the surrounding microenvironment for the colonization of circulating tumor cells at distant metastatic sites and before metastasis and spreading.The formation of PMNs includes processes such as vascular leakage and permeability changes,coagulation abnor-malities,extracellular matrix remodeling,migration and recruit-ment of bone marrow-derived cells,and immunosuppression,in which tumor-derived exosomes,tumor-derived soluble factors(TDSFs),bone marrow-derived cells(BMDCs),etc.are crucial in this process.This article focuses on the pre-metastasis microen-vironment,in-depth analysis of representative factors such as tumor-derived secretory factors,exosomes,bone marrow-derived cells and myeloid-derived suppressor cells,and other molecular network mechanisms involved in the formation of PMNs,and introduces tumors The characteristics of the metastatic cascade and the selection of organ parts outline the different roles of related molecules in the regulation of metastasis.That provide an effective reference value in order to better understand the mechanisms of tumor metastasis,tumor metastasis look for potential markers and diagnosis and treatment of cancer.

Dynamically changing antineutrophil cytoplasmic antibodies in granulomatosis with polyangiitis:A case report

BACKGROUND Granulomatosis with polyangiitis(GPA)is one of the most prevalent forms of the antineutrophil cytoplasmic antibody(ANCA)-associated vasculitis.GPA is characterized histologically by necrotizing granulomatous inflammation in addition to vasculitis.The diagnosis of GPA depends on clinical presentation,serological evidence of a positive ANCA,and/or histological evidence of necrotizing vasculitis or granulomatous destructive parenchymal inflammation.Cytoplasmic ANCA(c-ANCA)is positive in 65%-75% of GPA patients,accompanied by proteinase 3(PR3),the main target antigen of c-ANCA,another 5% of GPA patients had negative ANCA.CASE SUMMARY The patient,a 52-year-old male,presented with unexplained nasal congestion,tinnitus,and hearing loss.After a duration of 4 months experiencing these symptoms,the patient subsequently developed fever and headache.The imaging examination revealed the presence of bilateral auricular mastoiditis and partial paranasal sinusitis,and the ANCA results were negative.The anti-infective therapy proved to be ineffective,but the patient's symptoms and fever were quickly relieved after 1 wk of treatment with methylprednisolone 40 mg once a day.However,after continuous use of methylprednisolone tablets for 3 months,the patient experienced a recurrence of fever accompanied by right-sided migraine,positive c-ANCA and PR3,and increased total protein in cerebrospinal fluid.The and cyclophosphamide 0.8 g monthly,the patient experienced alleviation of fever and headache.Additionally,the ANCA levels became negative and there has been no recurrence.CONCLUSION For GPA patients with negative ANCA,there is a potential for early missed diagnosis.The integration of histopathological results and multidisciplinary communication plays a crucial role in facilitating ANCA-negative GPA.

Total body irradiation of donors can alter the course of tolerance and induce acute rejection in a spontaneous tolerance rat liver transplantation model

Liver transplantation is an established therapy for end-stage liver diseases. Graft rejection occurs unless the recipient receives immunosuppression after transplantation. This study aimed to explore the mechanism of acute rejection of liver allografts in rats pre-treated with total body irradiation to eliminate passenger lymphocytes and to define the role of CD4＋CD25＋ regulatory T cells in the induction of immunotolerance in the recipient. Male Lewis rats were used as donors and male DA rats were re- cipients. Rats were randomly assigned to the following four groups： control group, homogeneity liver transplantation group, idio-immunotolerance group and acute rejection group. After transplantation, the survival time of each group, serum alanine aminotransferase, total bilirubin levels, number of Foxp3＋CD4＋CD25＋ regulatory T cells, expression of glucocorticoid-induced tumor necrosis factor receptor on T cell subgroups, histopathology of the hepatic graft and spleen cytotoxic T lymphocyte lytic activity were measured. In the acute rejection group, where donors were preconditioned with total body in＇adiation before liver transplantation, all recipients died between day 17 and day 21. On day 14, serum alanine aminotransferase increased signifi- cantly to （459.2±76.9） U L^- 1, total bilirubin increased to （124.1±33.7） μmol L-1 （P〈0.05） and the ratio of Foxp3＋CD4＋CD25＋ regulatory T cells decreased significantly to 1.50%±0.50% （P〈0.05） compared with the other groups. Analysis of the T cell subpopulations in the acute rejection group varied from the other groups. Histological analysis showed typical changes of acute rejection in the acute rejection group only. Preconditioning of the donors with total body irradiation eliminated passenger lymphocytes of the liver graft, and thus affected the course of tolerance and induced acute rejection after liver transplantation.

Polyglycolic Acid Fibrous Scaffold Improving Endothelial Cell Coating and Vascularization of Islet

Background：Improving islet graft revascularization has become a crucial task for prolonging islet graft survival.Endothelial cells （ECs） are the basis of new microvessels in an isolated islet,and EC coating has been demonstrated to improve the vascularization and survival of an islet.However,the traditional method of EC coating of islets has low efficiency in vitro.This study was conducted to evaluate the effect of a polyglycolic acid （PGA） scaffold on the efficiency of islet coating by ECs and the angiogenesis in the coated islet graft.Methods：A PGA fibrous scaffold was used for EC coating of islet culture and was evaluated for its efficiency of EC coating on islets and islet graft angiogenesis.Results：In in vitro experiments,we found that apoptosis index of ECs-coating islet in PGA group （27% ± 8%） was significantly lower than that in control group （83% ± 20%,P 〈 0.05） after 7 days culture.Stimulation index was significantly greater in the PGA group than in the control group at day 7 after ECs-coating （2.07 ± 0.31 vs.1.80 ± 0.23,P 〈 0.05）.vascular endothelial growth factor （VEGF） level in the PGA group was significantly higher than the coating in the control group after 7 days culture （52.10 ± 13.50 ng/ml vs.16.30 ± 8.10 ng/ml,P 〈 0.05）.Because of a tight,circumvallated,adhesive and three-dimensional growth microenvironment,islet cultured in a PGA scaffold had higher coating efficiency showing stronger staining intensity of enzyme than those in the control group after 14 days of culture following ECs-coating.For in vivo study,PGA scaffold significantly prolonged the average survival time of EC-coated islet graft after transplantation compared with control group （15.30 ± 5.60 days vs.8.30 ± 2.45 days,P 〈 0.05）.The angiogenesis and area of survived grafts were more in the PGA group compared with the control group by measuring the mean microvessel density （8.60 ± 1.21/mm2 vs.5.20 ± 0.87/mm2,P 〈 0.05）.In addition,expression of VEGF and tyrosin-protein kinase receptor （Tie-2） gene increased in PGA scaffold group than that in control group by real-time reverse transcription-polymerase chain reaction analysis.Conclusions：These results demonstrate that the efficiency of EC coating of islets was successfully increased by culturing ECs on a PGA scaffold.This method enhances the function,survival,and vascularization of isolated islets in vitro and in vivo.

Pluripotent Stem Cell-derived Strategies to Treat Acute Liver Failure: Current Status and Future Directions

Liver disease has long been a heavy health and economic burden worldwide.Once the disease is out of control and progresses to end-stage or acute organ failure,orthotopic liver transplantation(OLT)is the only therapeutic alternative,and it requires appropriate donors and aggressive administration of immunosuppressive drugs.Therefore,hepatocyte transplantation(HT)and bioartificial livers(BALs)have been proposed as effective treatments for acute liver failure(ALF)in clinics.Although human primary hepatocytes(PHs)are an ideal cell source to support these methods,the large demand and superior viability of PH is needed,which restrains its wide usage.Thus,a finding alternative to meet the quantity and quality of hepatocytes is urgent.In this context,human pluripotent stem cells(PSC),which have unlimited proliferative and differential potential,derived hepatocytes are a promising renewable cell source.Recent studies of the differentiation of PSC into hepatocytes has provided evidence that supports their clinical application.In this review,we discuss the recent status and future directions of the potential use of PSC-derived hepatocytes in treating ALF.We also discuss opportunities and challenges of how to promote such strategies in the common applications in clinical treatments.

Association between the eNOS gene polymorphisms and rheumatoid arthritis risk in a northern Chinese population

Background Several genetic polymorphisms in the endothelial nitric oxide synthase （eNOS） gene are associated with the pathogenesis of rheumatoid arthritis （RA）. The objective of the present study was to investigate whether the two SNPs （T-786C and G894T） of the eNOS gene are associated with rheumatoid arthritis risk in a northern Chinese population. Methods In this study, the eNOS genes T-786C and G894T were studied in 196 cases with rheumatoid arthritis and 201 healthy controls with gender, age and ethnicity matched. The two SNPs were genotyped using polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. The analyses of association were statistically compared using the chi-square test with SPSS software for Windows. Results The frequency of the -786C allele was significantly higher in the rheumatoid arthritis patients than in the healthy controls （19.64% vs. 14.18%, P 〈0.05）. However, the 894T allele of the eNOS gene was not increased in the rheumatoid arthritis patients compared to the healthy controls. Conclusions Individuals with the -786CC genotype have an increased risk of rheumatoid arthritis. Further study with an increased sample size is necessary for the study of the role of this SNP in rheumatoid arthritis.

Whole-body magnetic resonance imaging vs.clinical evaluation of enthesitis in patients with spondyloarthritis

To the Editor:Enthesitis is an inflammation of the iliac attachment of ligaments or tendons.In spondyloarthropathies,peripheral enthesitis precedes joint symptoms and is associated with a higher degree of erosive disease.Four methods are available for evaluation of enthesitis,including tenderness,conventional radiography,ultrasonography,and magnetic resonance imaging(MRI).However,over 100 entheses are found in the human body.

Current health resources required for the management of ankylosing spondylitis in developing areas of China

To the Editor:Ankylosing spondylitis(AS)is a chronic inflammatory disease that causes bony fusion of spine and sacroiliac joints,resulting in restricted spinal movement and deformity.In China,the prevalence of AS is about 0.29%with a male to female ratio of approximately 3:1,and most cases occur in the adolescent period.[1]The misdiagnosis rate of AS patients in China is>50%.

Shikonin alleviates collagen-induced arthritis mice by inhibiting M1 macrophage polarization

OBJECTIVE: To investigate the effects of shikonin(SKN) on M1 and M2 polarization of macrophages both in vitro and in vivo. METHODS: Collagen-induced arthritis(CIA) in male DBA/1 mice were treated with a dose of 4 mg/kg/day of SKN for 23 d(n = 6/group). The histopathology of inflamed joints in CIA mice was evaluated to test the antiarthritic effect of SKN. M1/M2 polarization of macrophages induced by lipopolysaccharide(LPS) and interferon(IFN)-γ or interleukin(IL)-4 and IL-13, were used to assess the effect of SKN(0.05, 0.1, and 0.2 μM). The effect of SKN on the protein expression of nitric oxide synthase, arginase, CD68, and CD206 was evaluated using western blot analysis. RESULTS: The results of this study revealed that SKN delayed the arthritis feet symptom score, reduced the incidence rate of arthritis, and relieved the inflammation of joints in CIA mice. SKN inhibited M1 macrophage polarization but did not affect M2 macrophage polarization in the joints of CIA mice. Moreover, SKN inhibited M1 polarization induced by LPS and IFN-γ, but did not affect M2 polarization induced by IL-4 and IL-13. CONCLUSION: These findings suggest that SKN alleviated CIA through inhibiting M1 macrophage polarization and has great potential as a new drug for RA treatment.