Possible effects of chemokine-like factor-like MARVEL transmembrane domain-containing family on antiphospholipid syndrome.

ABSTRACT: Antiphospholipid syndrome (APS) is a systemic autoimmune disease defined by thrombotic or obstetrical events and persistent antiphospholipid antibodies (aPLs). Chemokine-like factor-like MARVEL transmembrane domain-containing family (CMTM) is widely expressed in the immune system and may closely related to APS. This review aimed to systematically summarize the possible effects of CMTM on APS. Publications were collected from PubMed and Web of Science databases up to August 2020. CKLF, CKLFSF, CMTM, antiphospholipid syndrome, immune cells, and immune molecules were used as search criteria. Immune cells, including neutrophil, dendritic cells (DCs), T-cells, B-cells, and inflammatory cytokines, play an important role in the development of APS. Chemokine-like factor 1 (CKLF1) has a chemotactic effect on many cells and can affect the expression of inflammatory cytokines and adhesion molecules through the nuclear factor-kB (NF-kB) pathway or mitogen-activated protein kinase (MARK) pathway. CKLF1 can participate in the maturation of DCs, T lymphocyte activation, and the activation of neutrophils through the MAPK pathway. CMTM1 may act on Annexin A2 by regulating Ca2+ signaling. CMTM2 and CMTM6 are up-regulated in neutrophils of APS patients. Some CMTM family members influence the activation and accumulation of platelets. CMTM3 and CMTM7 are binding partners of B-cell linker protein (BLNK), thereby linking B cell receptor (BCR) and activating BLNK-mediated signal transduction in B cells. Moreover, CMTM3 and CMTM7 can act on DCs and B-1a cell development, respectively. CMTM may have potential effects on the development of APS by acting on immune cells and immune molecules. Thus, CMTM may act as a novel prognostic factor or immunomodulatory treatment option of APS.

Dihydroartemisinin Inhibits the Proliferation, Colony Formation and Induces Ferroptosis of Lung Cancer Cells by Inhibiting PRIM2/SLC7A11 Axis.

OBJECTIVE: Lung cancer is the first leading cause of cancer-related deaths both worldwide and in China and threatens human health and quality of life. New drugs and therapeutic methods are urgently needed. Our study evaluated the roles of dihydroartemisinin (DHA) in lung cancer and further explored its underlying mechanisms. METHODS: CCK-8, colony formation and trypan blue exclusion assays were used to detect the cell viability, colony formation ability and cell death. qRT-PCR and Western blotting assays were applied to analyze the expressions of key molecules. RESULTS: DHA inhibited the proliferation and colony formation abilities and enhanced the cell death and induced ferroptosis of lung NCI-H23 and XWLC-05 cancer cells. DHA reduced PRIM2 expression and silencing PRIM2 mimicked the inhibitory roles on proliferation and colony formation and promotive roles on cell death and ferroptosis of DHA in lung NCI-H23 and XWLC-05 cancer cells. We further found that DHA treatment and loss of PRIM2 reduced the GSH level and increased the cellular lipid ROS and mitochondrial MDA levels, and further downregulated the expressions of SLC7A11 and ß-catenin in lung cancer cells, respectively. Exogenetic overexpression of PRIM2 recovered the inhibitory effects of DHA on proliferation and colony formation in lung NCI-H23 cancer cells, meanwhile loss of PRIM2 sensitizes NCI-H23 cells to DHA therapy. In vivo experiment further showed that DHA treatment significantly suppressed the tumor growth and downregulated PRIM2 and SLC7A11. CONCLUSION: Our study suggested that DHA inhibited the proliferation, colony formation and enhanced cell death and induced ferroptosis of lung cancer cells by inactivating PRIM2/SLC7A11 axis. Loss of PRIM2 induced ferroptosis might developed to be a novel therapeutic method in lung cancer therapy.

Change of serum B-cell activating factor level in patients with positive antiphospholipid antibodies and previous adverse pregnancy outcomes and its significance.

BACKGROUND: B-cell activating factor (BAFF) is vital for B cell survival. Serum BAFF levels are elevated in thrombotic antiphospholipid syndrome, but little is known about levels in patients with positive antiphospholipid antibodies (aPLs) and previous adverse pregnancy outcomes (APOs). We aimed to analyze serum BAFF concentrations of these patients in early pregnancy along with different pregnancy outcomes. METHODS: Thirty-six pregnant patients positive for aPLs and previous APOs (patient group), 25 healthy pregnant females (HP group) and 35 healthy non-pregnant females (HNP group) from the Peking University Third Hospital, between October 2018 and March 2019, were enrolled in this study. Serum of HNP and serum of patients as well as HP in the first gestational trimester were collected. Enzyme-linked immunosorbent assay kits were used to measure serum BAFF and interferon-alpha (IFN-α) concentrations. Cytometric bead array analysis was used to measure serum concentrations of cytokines. The patient group was further divided into APOs and non-APOs (NAPOs) group, fetal loss and live birth group according to pregnancy outcomes. The Mann-Whitney U-test was used to assess significance between and within groups. Spearman rank-order was used to evaluate correlation coefficients between BAFF and related cytokines. RESULTS: The serum BAFF level in HP group was significantly lower than HNP group (245.24 [218.80, 265.90] vs. 326.94 [267.31, 414.80] pg/mL, Z = -3.966, P < 0.001). The BAFF level was obviously elevated in patient group compared to that in HP group (307.77 [219.86, 415.65] vs. 245.24 [218.80, 265.90] pg/mL, Z = -2.464, P = 0.013). BAFF levels in APOs group tended to be higher than that in NAPOs group (416.52 [307.07, 511.12] vs. 259.37 [203.59, 375.81] pg/mL, Z = -2.718, P = 0.006). Compared to HP group, concentrations of IFN-α, interleukin (IL-6) and tumor necrosis factor were higher in patient group (33.37 [18.85, 48.12] vs. 13.10 [6.85, 25.47] pg/mL, Z = -2.023, P = 0.043; 39.16 [4.41, 195.87] vs. 3.37 [2.92, 3.90] pg/mL, Z = -3.650, P < 0.001; 8.23 [2.27, 64.46] vs. 1.53 [1.25, 2.31] pg/mL, Z = -3.604, P < 0.001, respectively). Serum BAFF levels had a positive correlation with the concentrations of both IL-6 and IL-10 (IL-6: r = 0.525, P = 0.002; IL-10: r = 0.438, P = 0.012). CONCLUSIONS: Serum BAFF levels are increased in patients with positive aPLs and previous APOs as compared to healthy pregnant females and tend to be higher in individuals with current APOs. The BAFF levels have a positive correlation with serum IL-6 and IL-10.

Chemokine-like factor-like MARVEL transmembrane domain-containing family in autoimmune diseases.

The chemokine-like factor (CKLF)-like MARVEL transmembrane domain-containing family (CMTM) is widely expressed in the immune system. Abnormal expression of CMTM is associated with the development of various diseases. This article summarizes the relevant research on the role of the CMTM family in immune disorders. This information will increase our understanding of pathogenesis and identify promising targets for the diagnosis and treatment of autoimmune diseases. The CMTM family is highly expressed in peripheral blood mononuclear cells. CKLF1 may be involved in the development of arthritis through its interaction with C-C chemokine receptor 4. CKLF1 is associated with the pathogenesis of lupus nephritis and psoriasis. Both CMTM4 and CMTM5 are associated with the pathogenesis of systemic lupus erythematosus. CMTM1, CMTM2, CMTM3, and CMTM6 play a role in rheumatoid arthritis, systemic sclerosis, Sjögren syndrome, and anti-phospholipid syndrome, respectively. The CMTM family has been implicated in various autoimmune diseases. Further research on the mechanism of the action of CMTM family members may lead to the development of new treatment strategies for autoimmune diseases.

CXCL16 upregulates RANKL expression in rheumatoid arthritis synovial fibroblasts through the JAK2/STAT3 and p38/MAPK signaling pathway.

OBJECTIVE: To explore the influence of chemokine, CXCL16, on the expression of the receptor activator nuclear factor κB ligand (RANKL) in rheumatoid arthritis (RA) fibroblast-like synoviocytes (RA-FLS). METHODS: The expression of CXCL16/CXCR6 and RANKL in RA or osteoarthritis (OA) patient synovia was examined by Western blot and immunohistochemistry. The serum concentration of CXCL16 and RANKL was measured by enzyme-linked immunosorbent assay (ELISA). RA-FLS were treated with recombinant CXCL16, and RANKL mRNA and protein were measured using PCR, Western blot and ELISA. RESULTS: The synovial expression of CXCL16, CXCR6, and RANKL was higher in RA patients than in patients with OA. The serum CXCL16 and RANKL levels were higher in RA patients compared with OA patients and healthy controls. CXCL16 correlated with erythrocyte sedimentation rate, C reactive protein, disease activity, serum rheumatoid factor, and RANKL. RA-FLS treated with CXCL16 showed markedly increased expression of RANKL. When STAT3 or p38 activation was blocked by an inhibitor, CXCL16 failed to upregulate RANKL expression. In contrast, inhibiting the Akt or Erk pathway did not achieve the same effect. CONCLUSIONS: CXCL16 upregulates RANKL expression in RA-FLS and these effects are mainly mediated by the JAK2/STAT3 and p38/MAPK signaling pathways.

Efficacy of low-dose 90Sr-90Y therapy combined with topical application of 0.5% timolol maleate solution for the treatment of superficial infantile hemangiomas.

The aim of the present study was to investigate the clinical efficacy and safety of low-dose 90Sr-90Y therapy combined with the topical application of 0.5% timolol maleate solution for the treatment of superficial infantile hemangiomas (IHs). A total of 72 infants with hemangiomas were allocated at random into the observation group (17 cases aged ≤3 months, 20 cases aged >3 months) or the control group (15 cases aged ≤3 months, 20 cases aged >3 months). The observation group was treated with low-dose 90Sr-90Y combined with timolol, while the control group received an identical dose of 90Sr-90Y with physiological saline. Data were collected for statistical analysis, and treatment efficacy was compared between the two groups. In the observation group, 100% (37/37) of subjects exhibited an 'excellent' response to the treatment, while 94.1% (16/17) of patients aged ≤3 months and 85.0% (17/20) aged >3 months were classed as being cured. In the control group, the treatment was classed as 'effective' in 100% (35/35) of the subjects, while the excellent response rate was 86.7% (13/15) among the infants aged ≤3 months and 75.0% (15/20) among the infants aged >3 months. The 'cure' rates in the control group were 66.7% (10/15) and 60.0% (12/20) for the ≤3-month- and >3-month-old subjects, respectively. The excellent response and cure rates were notably higher in the observation group than those in the control group. Comparison between the two groups revealed a χ2 value of 13.90 (P<0.01) for excellent responses in subjects aged ≤3 months, while for patients aged >3 months the χ2 value was 28.57 (P<0.01). Analysis of the cure responses gave similar results [≤3 months, χ2=23.22 (P<0.01); >3 months, χ2=15.67 (P<0.01)]. At 3-4 months after the first course of treatment, the cure rate was 33.3% (11/33) in the observation group, which was significantly higher than the rate of 18.32% (4/22) in the control group (χ2=5.92, P<0.05). No serious adverse reactions were observed in either group. In summary, low-dose 90Sr-90Y therapy combined with the topical application of 0.5% timolol maleate induces a rapid response in superficial IH, with excellent efficacy and no obvious adverse reactions, and may represent a clinically applicable intervention.

[Pregnancy outcome in 54 patients with antiphospholipid syndrome: a retrospective clinical study].

OBJECTIVE: To summarize the maternal/fetal outcome of pregnancy in antiphospholipid syndrome (APS) patients to evaluate the influence of treatment on the outcomes of pregnancy, and to investigate the possible clinical predictors of unsuccessful pregnancy. METHODS: The clinical characteristics, laboratory profiles and the outcomes of delivery of 54 APS patients from January 2000 to March 2013 were investigated retrospectively. RESULTS: (1) Maternal/fetal outcome: 17 pregnancies (31.4%) resulted in full term delivery, 7 (12.9%) in stillbirth, 16 (29.6%) in spontaneous abortion,10 (18.5%) in premature birth due to eclampsia or severe preeclampsia or signs of placental insufficiency, 4 (7.4%)received therapeutic termination of pregnancy due to eclampsia or severe preeclampsia. In 27 live birth cases, 8 (29.6%) were fetal growth restriction, 4 (14.8%) were low birth weight infants, and 3 (11.1%) were very low birth weight infants. (2) Influence of treatment on the pregnancy outcomes and complications: 24 APS patients were given the treatment of aspirin or aspirin combined with low molecular weight heparin, and 30 patients received no treatment. Compared with the untreated group, the treated group had lower rate of fetal loss, higher rate of full-term delivery, increased gestational age and birth weight, decreased incidence of preeclampsia / eclampsia and thrombocytopenia. There was a significant difference between the two groups (P<0.05). (3)Possible risk factors of unsuccessful pregnancy: there were 17 successful pregnancies and 37 unsuccessful pregnancy. The rate of double APL positive and antibody titers ≥ three times the upper limit of normal were higher in the unsuccessful pregnancy group than the successful pregnancy group. Antibody negative rate before pregnancy proportion of patients received treatment and the level of complement 4 were lower in the unsuccessful pregnancy group. CONCLUSION: Pregnant women with APS are an extremely high risk group for adverse maternal /fetal outcome. Treatments can improve the pregnancy outcome of the APS patients. APL not turning negative before pregnancy double APL positive, antibody titers ≥ three times the upper limit of normal and complement 4 decrease may be the risk factors for pregnancy failure and treatment may be a protective factor for successful pregnancy.

AG490 inhibits NFATc1 expression and STAT3 activation during RANKL induced osteoclastogenesis.

Commonly, JAK/STAT relays cytokine signals for cell activation and proliferation, and recent studies have shown that the elevated expression of JAK/STAT is associated with the immune rejection of allografts and the inflammatory processes of autoimmune disease. However, the role which JAK2/STAT3 signaling plays in the receptor activator of nuclear factor-κB ligand (RANKL)-mediated osteoclastogenesis is unknown. In this study, we investigated the effects of AG490, specific JAK2 inhibitor, on osteoclast differentiation in vitro. AG490 significantly inhibited osteoclastogenesis in murine osteoclast precursor cell line RAW264.7 induced by RANKL. AG490 suppressed cell proliferation and delayed the G1 to S cell cycle transition. Furthermore, AG490 also suppressed the expression of nuclear factor of activated T cells (NFAT) c1 but not c-Fos in RAW264.7. Subsequently, we investigated various intracellular signaling components associated with osteoclastogenesis. AG490 had no effects on RANKL-induced activation of Akt, ERK1/2. Interestingly, AG490 partly inhibited RANKL-induced phosphorylation of Ser(727) in STAT3. Additionally, down-regulation of STAT3 using siRNA resulted in suppression of TRAP, RANK and NFATc1 expression. In conclusion, we demonstrated that AG490 inhibited RANKL-induced osteoclastogenesis by suppressing NFATc1 production and cell proliferation via the STAT3 pathway. These results suggest that inhibition of JAK2 may be useful for the treatment of bone diseases characterized by excessive osteoclastogenesis.

[Analysis for clinical features of cardiovascular complication of diffuse connective tissue disease].

OBJECTIVE: To analyze the clinical features and prognosis of cardiovascular complications of diffuse connective tissue disease(dCTD). METHODS: Clinical data of cardiovascular complications of 181 cases of dCTD were retrospectively analyzed. Clinical data of two subsets [rheumatoid arthritis (RA, 81) and systemic lupus erythematosus (SLE, 42)] were also analyzed. Follow-up was carried out for all the patients. RESULTS: RA and SLE were the most common disease complicated by cardiovascular disease needing hospitalization. The most frequent cardiovascular complications in dCTD patients were hypertension, hyperlipidemia and coronary artery disease. The most common echocardiographic abnormalities were valvular regurgitation, left ventricular diastolic dysfunction, enlargement of left atrium, pulmonary hypertension and pericardial effusion (57.8%, 50.6%, 33.7%, 21.7% and 19.3%, respectively). Compared with the subset of RA, patients were younger at onset of hypertension, coronary artery disease and hyperlipidemia in the subset of SLE [(40+/-11) vs (56+/-15), P<0.001; (53+/-12) vs (64+/-10), P=0.011; (44+/-16) vs (58+/-12), P=0.012, respectively]. Both pericardial effusion (P<0.001) and enlargement of left ventricle (P=0.03) were more frequent, and left ventricular diastolic dysfunction was less common (P<0.001). Median survival time of these dCTD patients was 9.8 years. CONCLUSION: RA and SLE are the most common diffuse connective tissue diseases complicated by cardiovascular disease needing hospitalization. The most frequent cardiovascular complications in dCTD patients are hypertension, hyperlipidemia and coronary artery disease. The prognosis of dCTD patients complicated with cardiovascular diseases is poor. SLE patients are younger at onset of cardiovascular diseases.

Comparative study on circulating endothelial progenitor cells in systemic lupus erythematosus patients at active stage.

Circulating endothelial progenitor cells (CEPCs) play an important role in the process of atherosclerosis. Most previous studies on CEPC in systemic lupus erythematosus (SLE) patients were on their number and some functions and the results were not consistent. No studies on their anti-inflammatory function and integrated status were reported. The purpose of this study was to determine the number, function (including anti-inflammatory function), and the integrated status of CEPCs in active SLE patients. The study was performed in 35 active SLE patients (28 females, 7 males) and 35 age-and gender-matched healthy controls. CEPC number was determined by Fluorescence-Activated Cell Sorting. Proliferation capacity of CEPC was assessed by PCNA staining. Adhesion capacity of CEPC to fibronectin and adhesion capacity of THP1 cell to CEPC were determined by cell adhesion assay. Migratory capacity of CEPC was measured by transwell chamber assay and the potential to form tubes on Matrigel of CEPC was determined by in vivo tube formation on Matrigel test. The expression of inducible nitric oxide synthase (iNOS) and interleukin-6 (IL-6) assessed by quantitative PCR as well as the expression of intercellular adhesion molecule-1 (ICAM-1) and phosphorylated-Akt (p-Akt) assessed by western-blotting were used to evaluate the anti-inflammatory function and cell status of CEPCs. The number of CEPC in SLE patients was not different from that in control (p > 0.05). Proliferation capacity of CEPC was decreased in active SLE patients (p = 0.027). Adhesion capacity of CEPC to fibronectin was decreased (p = 0.04) in SLE patients and adhesion capacity of THP1 cell to CEPC was increased in SLE patients (p < 0.001). Migratory activity was reduced in patient CEPCs (p < 0.001). Capacity of CEPCs to form tube on Matrigel was decreased in SLE patients (p < 0.001). Expression of iNOS and IL-6 (p < 0.001, p = 0.006, respectively) and ICAM-1 were increased in CEPC of SLE patients and expression of p-Akt was decreased in CEPC of SLE patients. Our data show that CEPC number in active SLE patients was not significantly different from healthy controls, but their functions were partly impaired, including proliferation, adhesion, migration, and tube formation. Bad cell status and increased susceptibility to inflammatory process of CEPCs in active SLE were also observed in our study.

Comparative study on low back pain misdiagnosed as spondyloarthropathy.

This study aims to investigate features of different diseases with low back pain misdiagnosed as spondyloarthropathy so as to improve the accuracy of diagnosis for spondyloarthropathy. The clinical and laboratory data of 24 cases misdiagnosed as spondyloarthropathy in recent 3 years were comparatively and retrospectively analyzed. The diagnostic accuracy of the European Spondyloarthropathy Study Group (ESSG) criteria, Amor criteria, and the combination of them in these misdiagnosed cases was also analyzed. The final diagnoses of these 24 cases were listed below: four malignant tumors (retroperitoneal adipose sarcoma, advanced gastric carcinoma, ovarian papillary epithelioma, acute lymphocytic leukemia), six benign tumors (two parathyroid adenoma with hyperparathyroidism, one intraspinal lipoma, intraspinal ependymomas, sacral tubulocyst, and intraspinal schwannoglioma, respectively). The other 14 cases included fibromyalgia syndrome (3), osteitis condensans (3), diffuse idiopathic skeletal hyperostosis (2), lumbar intervertebral disk protrusion (1), congenital scoliosis (1), Wilson's disease (1), ochronosis (1), Fanconi syndrome (1) and hypophosphatemic rachiopathy (1). Among patients with tumor, all except three patients had persistent low back pain without morning stiffness, which aggravated at night and could not be relieved by rest or exercise. The symptoms could not be relieved by administration of multiple nonsteroidal anti-inflammatory drugs. Eleven patients had inflammatory low back pain defined by Calin. Of the total misdiagnosed cases, 54.17-83.33% could be prevented by application of ESSG criteria or Amor criteria, or a combination of them. From the data, we could see that the clinical features of different diseases with low back pain were different from each other and from those of spondyloarthropathy. The various criteria for spondyloarthropathy may be more effective in combination, along with other clinical information like these clinical features.

Less disease severity and favorable prognosis are associated with postmenopausal systemic lupus erythematosus patients.

The aim of this study is to characterize the clinical manifestations of postmenopausal systemic lupus erythematosus (SLE) patients. Of the 699 SLE inpatients, 20 postmenopausal and 70 menstruous SLE patients were evaluated and compared for the clinical manifestations. The mean age of onset was 55.05 years (range from 42 to 66) with a peak of 50-60 in postmenopausal lupus patients. The average time from SLE onset to diagnosis was 2.18 years. Arthritis was the most frequent initial manifestation in the postmenopausal group. Other common clinical manifestations and laboratory abnormalities include lassitude, fever, alopecia, malar rash, cardiac impairment and weight loss, and elevated ESR, decreased C3, ANA >or= 1:80, hypergammaglobulinemia and anti-RNP antibody positive. Compared with menstruous lupus patients, postmenopausal patients were more likely to have weight loss (P < 0.01), myalgia and myasthenia (P < 0.01), and less likely to have malar rash (P < 0.05), renal involvement (P < 0.01), leukocytopenia (P < 0.05) and positive ANA (P < 0.01). Thus, less disease severity and favorable prognosis were associated with postmenopausal SLE patients. Misdiagnosis and missed diagnosis were easy to make with their non-specific symptoms with fewer features suggestive of diagnosis.

[Recombinant adeno-associated viral vector 2-mediated gene transfer of VEGF165 improved porcine chronic myocardial ischemia].

OBJECTIVE: Vectors commonly used for therapeutic angiogenesis such as adenovirus and plasmid had their own limitations. Adenovirus-associated virus (AAV) is a relatively new but probably more ideal vector as it is safe and efficient. We will study the efficiency of recombinant AAV-2 mediated vascular endothelial growth factor165 gene transfer in inducing angiogenesis and arteriogenesis, in improving blood flow and myocardium function in a porcine chronic myocardial ischemic model. METHODS: Chinese experimental minipigs underwent placement of a left circumflex artery aneroid constrictor. Five weeks later, electrocardiogram, coronary angiography and magnetic resonance imaging were performed to confirm occlusion of LCX or ischemia of myocardium in LCX territory. Coronary blood flow, myocardium perfusion and left ventricular wall function were also evaluated. Then the animals were randomized to treatment with rAAV2-VEGF(165) (1 x 10(12) virus genome) or administration of PBS, both by direct myocardial injection. Three and six months after therapy, the animals were evaluated with regard to expression of VEGF(165) Capillary density and arteriole density of the ischemic myocardium, coronary angiography, myocardial perfusion and left ventricular function were also assessed six months after therapy. RESULTS: Five weeks after aneroid occluder implantation, all the animals demonstrated complete or nearly complete occlusion of LCX and perfusion deficiency in LCX territory. Three months after therapy, expression of VEGF(165) mRNA and protein were higher in the VEGF than control group. The difference between the two groups diminished after six months. There was significant increase in capillary density (1404.06 +/- 250.48/mm(2) vs 976.88 +/- 344.79/mm(2), P < 0.05) and arteriole density (167.81 +/- 36.29/mm(2) vs 116.56 +/- 34.48/mm(2), P < 0.05) in VEGF group compared with control. Comparison of myocardial perfusion demonstrated marked differences between the two groups with significant improvement in animals treated with rAAV2-VEGF(165). No significant improvement in left ventricular function was seen in either the VEGF or control group. CONCLUSIONS: Transmyocardial delivery of rAAV2-VEGF(165) resulted in VEGF gene expression for at least three months and stimulated angiogenesis and arteriogenesis in porcine model of chronic myocardial ischemia. Myocardial perfusion was also improved after VEGF gene delivery.