Efficacy of Botulinum Toxin B Injection for Raynaud's Phenomenon and Digital Ulcers in Patients with Systemic Sclerosis.

The efficacy and safety of botulinum toxin B (BTX-B) for treatment of Raynaud's phenomenon and digital ulcers in patients with systemic sclerosis was assessed. A total of 45 patients with systemic sclerosis who had Raynaud's phenomenon were blinded and divided randomly into 4 groups: a no-treatment control group, and 3 treatment groups, using 250, 1,000 or 2,000 international units (U) of BTX-B injections in the hand with more severe symptoms. Four weeks after injection, pain/numbness visual analogue scale scores and Raynaud's score in the groups treated with 1,000 and 2,000 U BTX-B were significantly lower than in the control group and the group treated with 250 U BTX-B. These beneficial effects were sustained until 16 weeks after the single injection. At 4 weeks after injection skin temperature recovery in the group treated with 2,000 U BTX-B was significantly improved. The numbers of digital ulcers in the groups treated with 1,000 and 2,000 U BTX-B were significantly lower than in the control group. In conclusion, 1,000 and 2,000 U BTX-B injections significantly suppressed the activity of Raynaud's phenomenon and digital ulcers in patients with SSc without serious adverse events.

Drug-induced Steatohepatitis Caused by Long-term Use of Topical Steroids for Atopic Dermatitis: A Case Report.

Atopic dermatitis is common in children and often treated with topical corticosteroids (TCs). A boy in his late teens who had been using TCs for atopic dermatitis was diagnosed with liver damage during a health checkup. A medical examination revealed severe steatotic liver disease and elevated liver enzyme levels despite the absence of typical symptoms such as central obesity. After discontinuation of TCs, an improvement in liver enzyme levels was observed, leading to the diagnosis of drug-induced steatohepatitis. This case underscores the potential liver risks associated with prolonged TC use in children, highlighting the need for parental education.

Anti-polymyositis/Scl antibody-positive overlap syndrome of diffuse cutaneous systemic sclerosis, dermatomyositis, systemic lupus erythematosus, and antiphospholipid syndrome.

A 37-year-old Japanese man with a 3-year history of diffuse cutaneous systemic sclerosis was admitted to our hospital with high fever, arthralgia, myalgia, and muscle weakness. A physical examination revealed facial erythema, Gottron's sign, and mechanic's hands in addition to skin sclerosis. Laboratory data revealed significantly elevated levels of creatine kinase and decreased complement. Anti-RNP, anti-Smith, anti-DNA, anti-ß2 -glycoprotein 1, anti-polymyositis (PM)/Scl75, and anti-PM/Scl100 antibodies were detected. He also had urinary protein, interstitial lung disease, pericarditis, multifocal cerebral infarctions, and leukoencephalopathy. Thus, a diagnosis of overlap syndrome of diffuse cutaneous systemic sclerosis, dermatomyositis, and systemic lupus erythematosus with antiphospholipid syndrome was made. Because of the intractable course, he was treated with multiple immunosuppressive and immunomodulatory drugs, including three rounds of 1000 mg methylprednisolone pulse therapy. This is the first case report of anti-PM/Scl antibody-positive overlap syndrome of three major connective tissue diseases.

Patient- and Therapist-Related Risk Factors for Adverse Events in Acupuncture and Moxibustion in Japan: Multicenter Survey of Acupuncture and Moxibustion Teaching Clinics.

Background/Objective: To investigate the patient- and therapist-related factors underlying adverse events (AEs) in acupuncture and moxibustion (A&M). Design: Retrospective study using data from medical records. Subjects: Patients who underwent A&M therapy in 4 clinics providing A&M over a 6-month period and their therapists. Main Outcome Measures: Survey items included the number of patients, age, sex, number of sessions, number and type of AEs, patients' underlying disease, and the therapist's years of clinical experience. The chi-squared test was used for intergroup comparisons. Spearman's rank correlation coefficient was used to analyze the correlations between the number of sessions and AEs. Logistic regression analysis was performed with AEs as the objective variable to investigate the relationships between the various parameters and AEs. Results: The analysis included 615 patients and 113 therapists. A total of 421 AEs occurred in a total of 4,369 sessions (9.6%). The number of sessions and number of AEs were significantly and positively correlated with patients (r = 0.47, P < 0.001) and therapists (r = 0.65, P < 0.001). Logistic analysis identified patient sex (adjusted odds ratio: 1.78, 95% confidence interval: [1.39-2.30]), liver disease (0.40, [0.19-0.84]), and years of clinical experience (to a cutoff of 2 or fewer years, 2-4 years: 0.65, [0.48-0.88], 5-9 years: 0.62, [0.44-0.87], 10 years or more: 0.50, [0.37-0.68]) as significant variables. Conclusions: Female sex and fewer years of clinical experience were factors that increased the risk of AEs, and underlying liver disease was a factor that decreased the risk of AEs.

Clustering of Lifestyle Factors and Its Association with Low Back Pain: A Cross-Sectional Study of Over 400,000 Japanese Adults.

PURPOSE: Although many studies have indicated the association between low back pain (LBP) and lifestyle factors, the combined effect of lifestyle factors on LBP has not been adequately investigated. We aimed to investigate the association between a cluster of unhealthy lifestyle behaviors and LBP using a large cohort of Japanese adults. METHODS: We included 419,003 adults aged over 20 years who underwent an annual health checkup between April 2013 and March 2014 in Japan. Information on the following lifestyle factors was collected using the standardized questionnaire: smoking, alcohol intake, exercise, physical activity, walking speed, weight control, eating habits, and sleep. Each factor was evaluated as a dichotomous variable (1: health risk, 0: no health risk). A lifestyle risk score was calculated by summing the score of each lifestyle factor (range: 0-12) and was categorized into three groups (low, moderate, high). LBP was defined as self-reported LBP under treatment. Logistic regression analysis was conducted to calculate the odds ratio (OR) and 95% confidence interval (CI) for LBP. RESULTS: In multivariable logistic regression analysis, the OR for LBP was significantly higher in the moderate-risk score group (adjusted OR: 1.33 [95% CI: 1.23-1.44] in men; 1.40 [95% CI: 1.27-1.54] in women) and the high-risk score group (adjusted OR: 1.54 [95% CI: 1.43-1.67] in men; 1.83 [95% CI: 1.64-2.03] in women) than in the low-risk score group. A trend of higher risk of LBP associated with higher lifestyle risk score was observed in both sexes (p for trend < 0.001). These results were similar even in subgroup analysis by age and body mass index (BMI). CONCLUSION: Clustering of unhealthy lifestyles was associated with increased risk of LBP regardless of age and BMI. These results may provide implications for better prevention and management of LBP, considering modifiable lifestyle factors.

Sex differences in the association of metabolic syndrome with low back pain among middle-aged Japanese adults: a large-scale cross-sectional study.

BACKGROUND: Although some recent studies have indicated an association between metabolic syndrome (MetS) and musculoskeletal disease, little is known about the association of MetS with low back pain (LBP). The present study aimed to investigate sex differences in the association of MetS and the clustering of MetS components with LBP among middle-aged Japanese individuals. METHODS: Study subjects were 45,192 adults (30,695 men, 14,497 women) aged 40-64 years who underwent annual health checkups conducted from April 2013 to March 2014. MetS was defined according to the criteria of the Examination Committee of Criteria for MetS in Japan as abdominal obesity plus at least two of dyslipidemia, high blood pressure, or high blood glucose. Information on LBP and health-related lifestyles were collected using a self-administered questionnaire. Logistic regression modeling was used to calculate the odds ratio (OR) and 95% confidence interval (CI) for LBP. RESULTS: After adjusting for age and lifestyle factors, the OR of MetS for LBP was 1.15 (95% CI 0.95-1.40) in men and 2.16 (95% CI 1.32-3.53) in women. Compared to subjects without abdominal obesity, the presence of abdominal obesity significantly increased the OR for LBP among men (abdominal obesity only: OR 1.34, 95% CI 1.02-1.76; abdominal obesity plus one component: OR 1.24, 95% CI 1.01-1.52; abdominal obesity plus two or more components: OR 1.26, 95% CI 1.02-1.55). Among women, adding other components of MetS to abdominal obesity significantly increased ORs for LBP (abdominal obesity only: OR 1.70, 95% CI 0.94-3.08; abdominal obesity plus one component: OR 1.66, 95% CI 1.06-2.60; abdominal obesity plus two or more components: OR 2.30, 95% CI 1.41-3.78). CONCLUSIONS: This large-scale cross-sectional study indicated that MetS was significantly associated with LBP among women only and that a sex-difference existed in the association between the clustering of MetS components and LBP. Clustering of MetS components by sex may need to be considered for the prevention of LBP, although further prospective studies are needed to clarify the causality.

Cutaneous adult xanthogranuloma with a small portion of BRAFV 600E mutated Langerhans cell histiocytosis populations: A case report and the review of published work.

Histiocytoses, including Langerhans cell histiocytosis (LCH), juvenile or adult xanthogranuloma (AXG) and Rosai-Dorfman disease (RDD), are rare disorders characterized by the proliferation of cells derived from monocyte/macrophage lineages. A few cases of LCH coexisting with xanthogranuloma or RDD have been reported. The etiology of these diseases remains unclear. However, oncogenic BRAFV 600E mutations have been identified in LCH. Here, we report the case of a 26-year-old Japanese man with a 3-month history of a solitary occipital nodule. No abnormality was detected in his other organs, and a total resection of the nodule was performed. Histopathological examination revealed the coexistence of LCH and AXG with prominent emperipolesis characteristic of RDD. Immunohistochemistry showed that most of the large histiocytes were positive for CD68, weakly positive or negative for S100, and negative for CD207 and CD1a, supporting the diagnosis of AXG. The tumor cells with emperipolesis did not show S100-positive findings characteristic of RDD. The focally aggregated oval histiocytic cells were positive for CD1a, CD207, CD68 and S100, and were compatible with the immunophenotype of LCH cells. In addition, these cells were positive for BRAFV 600E mutation. The tumor cells in our patient exhibited a cellular morphology characteristic of multiple histiocytoses in a solitary cutaneous nodule, which may imply an etiological association among LCH, AXG and RDD. To our knowledge, this is the first report of a BRAFV 600E mutation-positive case of LCH coexisting with AXG. Because patients with BRAFV 600E mutation have higher risks of multisystemic LCH and recurrence, we should carefully follow up the patient.

Suppressive Regulation by MFG-E8 of Latent Transforming Growth Factor ß-Induced Fibrosis via Binding to αv Integrin: Significance in the Pathogenesis of Fibrosis in Systemic Sclerosis.

OBJECTIVE: Several studies have demonstrated that the secreted glycoprotein and integrin ligand milk fat globule-associated protein with epidermal growth factor- and factor VIII-like domains (MFG-E8) negatively regulates fibrosis in the liver, lungs, and respiratory tract. However, the mechanisms and roles of MFG-E8 in skin fibrosis in systemic sclerosis (SSc) have not been characterized. We undertook this study to elucidate the role of MFG-E8 in skin fibrosis in SSc. METHODS: We assessed expression of MFG-E8 in the skin and serum in SSc patients. We examined the effect of recombinant MFG-E8 (rMFG-E8) on latent transforming growth factor ß (TGFß)-induced gene/protein expression in SSc fibroblasts. We examined the effects of deficiency or administration of MFG-E8 on fibrosis mouse models. RESULTS: We demonstrated that MFG-E8 expression around dermal blood vessels and the serum MFG-E8 level in SSc patients (n = 7 and n = 44, respectively) were lower than those in healthy individuals (n = 6 and n = 28, respectively). Treatment with rMFG-E8 significantly inhibited latent TGFß-induced expression of type I collagen, α-smooth muscle actin, and CCN2 in SSc fibroblasts (n = 3-8), which suggested that MFG-E8 inhibited activation of latent TGFß as well as TGFß signaling via binding to αv integrin. In a mouse model of bleomycin-induced fibrosis (n = 5-8) and in a TSK mouse model (a genetic model of SSc) (n = 5-10), deficient expression of MFG-E8 significantly enhanced both pulmonary and skin fibrosis, and administration of rMFG-E8 significantly inhibited bleomycin-induced dermal fibrosis. CONCLUSION: These results suggest that vasculopathy-induced dysfunction of pericytes and endothelial cells, the main cells secreting MFG-E8, may be associated with the decreased expression of MFG-E8 in SSc and that the deficient inhibitory regulation of latent TGFß-induced skin fibrosis by MFG-E8 may be involved in the pathogenesis of SSc and may be a therapeutic target for fibrosis in SSc patients.

The Regulation of Skin Fibrosis in Systemic Sclerosis by Extracellular ATP via P2Y2 Purinergic Receptor.

Tissue injury/hypoxia and oxidative stress induced-extracellular adenosine triphosphate (ATP) can act as damage-associated molecular pattern molecules, which initiate inflammatory response. Our objective was to elucidate the role of extracellular ATP in skin fibrosis in systemic sclerosis (SSc). We identified that hypoxia enhanced ATP release and that extracellular ATP enhanced IL-6 production more significantly in SSc fibroblasts than in normal fibroblasts. There were no significant differences of P2X and P2Y receptor expression levels between normal and SSc fibroblasts. Nonselective P2 receptor antagonist and selective P2Y2 receptor antagonists, kaempferol and AR-C118925XX, significantly inhibited ATP-induced IL-6 production and phosphorylation of p38 in SSc fibroblasts. ATP-induced IL-6 production was significantly inhibited by p38 inhibitors, SB203580, and doramapimod. Collagen type I production in SSc fibroblasts by ATP-induced IL-6/IL-6 receptor trans-signaling was inhibited by kaempferol and SB203580. The amount of ATP in bleomycin-treated skin was increased, and administration of AR-C118925XX significantly inhibited bleomycin-induced dermal fibrosis in mice. These results suggest that vasculopathy-induced hypoxia and oxidative stress might enhance ATP release in the dermis in SSc and that extracellular ATP-induced phosphorylation of p38 via P2Y2 receptor might enhance IL-6 and collagen type I production in SSc fibroblasts. P2Y2 receptor antagonist therapy could be a treatment for skin sclerosis in patients with SSc.

Inhibitory Regulation of Skin Fibrosis in Systemic Sclerosis by Apelin/APJ Signaling.

OBJECTIVE: Apelin/APJ signaling has been determined to regulate cardiac and arterial fibrosis and to be involved in the pathogenesis of pulmonary arterial hypertension. Our objective was to elucidate the role of apelin in skin fibrosis in systemic sclerosis (SSc). METHODS: Expression of apelin/APJ in normal and SSc fibroblasts was compared. Effects of small interfering RNA depletion and the addition of apelin in fibroblasts were analyzed. The effect of apelin injections on bleomycin-induced dermal fibrosis in mice was investigated. We analyzed the effects of the biased agonist of APJ, MM07, on skin fibrosis in vitro and in vivo. RESULTS: The expression of apelin in SSc fibroblasts was significantly lower than that in normal fibroblasts. Serum apelin levels were negatively correlated with the modified Rodnan skin thickness score in SSc patients. Stimulation with transforming growth factor ß1 (TGFß1) inhibited apelin expression in fibroblasts, suggesting that activation of TGFß1 signaling in SSc might be responsible for reduced apelin expression in SSc fibroblasts. Small interfering RNA depletion of apelin from fibroblasts significantly enhanced fibrosis-related gene expression, and treatment with apelin protein significantly inhibited TGFß1 signaling in fibroblasts. Administration of apelin significantly inhibited bleomycin-induced dermal fibrosis in mice. We demonstrated that MM07 had greater potential than apelin to inhibit fibrosis in vivo and in vitro. CONCLUSION: Collectively, TGFß1 signaling and apelin signaling may counteract each other in the fibrotic process of SSc. Inhibitory regulation of TGFß1-induced skin fibrosis by apelin/APJ signaling may be involved in the pathogenesis of SSc and could be a therapeutic target for fibrosis in SSc patients.

Botulinum toxin B suppresses the pressure ulcer formation in cutaneous ischemia-reperfusion injury mouse model: Possible regulation of oxidative and endoplasmic reticulum stress.

BACKGROUND: We previously identified that botulinum toxin A (BTX-A) suppressed pressure ulcer (PU) formation after cutaneous ischemia-reperfusion (I/R) injury; however, regulation of cutaneous I/R-induced oxidative and endoplasmic reticulum (ER) stress by BTX-B was not investigated. Additionally, the efficacy of BTX-B injection has never been examined. OBJECTIVE: Objective was to assess the effects of BTX-B on the formation of PU by cutaneous I/R injury, and the regulation of oxidative and ER stress in I/R injury by BTX-B. METHODS: BTX-B was subcutaneously injected into I/R area, and wound size, vascular damage, hypoxic area, and apoptotic cells in I/R area were analyzed. We evaluated the extent of oxidative and ER stress in I/R area by using OKD48 mice and ERAI mice, respectively, which enabled evaluating oxidative and ER stress through bioluminescence detection. RESULTS: BTX-B injection significantly suppressed the formation of PU by cutaneous I/R injury. Cutaneous I/R-induced vascular damage, hypoxic area, and number of oxidative-damaged cells and apoptotic cells were suppressed by BTX-B injection. BTX-B administration significantly inhibited I/R-induced oxidative stress signal in OKD48 mice. BTX-B reduced the I/R-induced oxidative stress-associated factors. BTX-B significantly inhibited the oxidant-induced reactive oxygen species and apoptosis of endothelial cells and fibroblasts. BTX-B significantly inhibited I/R-induced ER stress signal in ERAI mice. Cutaneous I/R injury-induced ER stress-response factors and GRP78/BiP and CHOP-positive cells in I/R area were significantly decreased by BTX-B injection. CONCLUSION: BTX-B injection might have protective effects against PU formation after cutaneous I/R injury by reducing vascular damage, hypoxia-induced oxidative and ER stress, and apoptosis.

Morphological classification of the anterior and middle talocalcaneal joints in terms of biomechanical stress dispersion.

Of the various foot joints, the level of interest shown in the talocalcaneal joint has been notably low. However, when considering stress dispersion during bipedal locomotion, the talocalcaneal joint plays an important role. Present study examined and morphologically classified the talocalcaneal joint. The present study investigated the morphology of the anterior talocalcaneal articular surface (A) and middle talocalcaneal articular surface (M) of the calcaneus. Using 94 feet of 47 human cadavers for anatomical dissection, the morphology of the A and M was classified into the following 6 types: Type 1, double ellipsoid, and A is markedly smaller than M; Type 2, gourd-form, and A is markedly smaller than M; Type 3, double ellipsoid, but A and M are partially in contact, Type 4, double ellipsoid, A and M are completely separated; Type 5, cylinder-form; and Type 6, others. Types 4 and 5 accounted for the majority of cases. In upright standing and bipedal locomotion, the anterior and middle talocalcaneal joints receive the anterior component of body weight dispersion in the posterior foot region. Morphological classification of articular surfaces that actually bear body weight is important, reflecting the degree of stress dispersion.

A Multicenter Prospective Survey of Adverse Events Associated with Acupuncture and Moxibustion in Japan.

Background: There have been only a few prospective surveys on adverse events (AEs) in Japanese-style acupuncture practice, and these surveys were conducted only in a single college acupuncture clinic. Objective: The goal of this research was to assess the safety of acupuncture and moxibustion performed in educational facilities in Japan. Materials and Methods: This was a multicenter prospective survey, using paper reporting forms. It was conducted in eight acupuncture clinics affiliated with educational institutions. The subjects were outpatients attending the clinics. The main outcome measure was the number of reported adverse events. The study was conducted for 5-7 months at each facility between October 2014 and June 2015. Participating acupuncture practitioners were instructed to self-report AEs observed during and after treatment; patients were interviewed upon treatment completion. For returning patients, treatment was preceded by an interview survey regarding the AEs identified after the previous treatment session. A specialized 4-sheet questionnaire was used. Results: Two hundred and thirty-two acupuncture practitioners participated, 2180 patients received treatment, and there were 14,039 sessions, overall. In total, 847 (6.03%) AEs were reported. The most common AEs included subcutaneous bleeding and hematomas (370, 2.64%), followed by discomfort (109, 0.78%) and residual pain at insertion points (94, 0.67%). No infections or serious AEs were reported. Conclusions: Acupuncture and moxibustion performed in educational facilities in Japan were safe because most of the AEs reported were mild and transient. However, the risk cannot be defined definitely because the survey sample size was too small.

Protective effect of mesenchymal stem cells on the pressure ulcer formation by the regulation of oxidative and endoplasmic reticulum stress.

Cutaneous ischemia-reperfusion (I/R) injury is associated with the early pathogenesis of cutaneous pressure ulcers (PUs). The objective of this study was to investigate the effect of mesenchymal stem cells (MSCs) injection on the formation of PUs after I/R injury and determine the underlying mechanisms. We found that the subcutaneous injection of MSCs into areas of I/R injured skin significantly suppressed the formation of PUs. I/R-induced vascular damage, hypoxia, oxidative DNA damage, and apoptosis were decreased by MSCs injection. Oxidative stress signals detected after I/R in OKD48 (Keap1-dependent oxidative stress detector, No-48-luciferase) mice were decreased by the injection of MSCs. In cultured fibroblasts, MSCs-conditioned medium significantly inhibited oxidant-induced reactive oxygen species (ROS) generation and apoptosis. Furthermore, endoplasmic reticulum (ER) stress signals detected after I/R in ERAI (ER stress-activated indicator) mice were also decreased by the injection of MSCs. These results suggest that the injection of MSCs might protect against the development of PUs after cutaneous I/R injury by reducing vascular damage, oxidative cellular damage, oxidative stress, ER stress, and apoptosis.

Mechanistic insight into the norepinephrine-induced fibrosis in systemic sclerosis.

Raynaud's phenomenon is frequently observed in systemic sclerosis (SSc) patients, and cold- or stress-induced norepinephrine (NE) has been speculated to be associated with vasoconstriction. Objective was to elucidate the role of NE in fibrosis in SSc. IL-6 is a potent stimulator of collagen production in fibroblasts. NE enhanced IL-6 production and proliferation more significantly in SSc fibroblasts than in normal fibroblasts. Furthermore, the production of IL-6 and phosphorylation of p38 in SSc fibroblasts was enhanced by adrenergic receptor (AR)ß agonist, isoproterenol, but not ARα agonist, oxymetazoline. ARß blocker, propranolol, inhibited NE-induced IL-6 production and phosphorylation of p38 in SSc fibroblasts. NE-induced IL-6 was significantly inhibited by p38 inhibitor, SB203580, suggesting that NE-induced phosphorylation of p38 via ARß enhances IL-6 production in SSc fibroblasts. NE-induced phosphorylation of ERK1/2 via ARα inhibited IL-6 production in SSc fibroblasts. Combined treatment with NE and endothelin-1 resulted in an additive increase in IL-6 production in SSc fibroblasts. NE-induced IL-6/IL-6 receptor trans-signaling increased the production of collagen type I in SSc fibroblasts, and both propranolol and SB203580 inhibited NE-induced collagen production. These results suggest that cold exposure and/or emotional stress-induced NE might contribute to the skin fibrosis via potentiation of IL-6 production from fibroblasts in SSc.

MFG-E8 Drives Melanoma Growth by Stimulating Mesenchymal Stromal Cell-Induced Angiogenesis and M2 Polarization of Tumor-Associated Macrophages.

Secretion of the powerful angiogenic factor MFG-E8 by pericytes can bypass the therapeutic effects of anti-VEGF therapy, but the mechanisms by which MFG-E8 acts are not fully understood. In this study, we investigated how this factor acts to promote the growth of melanomas that express it. We found that mouse bone marrow-derived mesenchymal stromal cells (MSC) expressed a substantial amount of MFG-E8. To assess its expression from this cell type, we implanted melanoma cells and MSC derived from wild type (WT) or MFG-E8 deficient [knockout (KO)] into mice and monitored tumor growth. Tumor growth and M2 macrophages were each attenuated in subjects coimplanted with KO-MSC compared with WT-MSC. In both xenograft tumors and clinical specimens of melanoma, we found that MFG-E8 expression was heightened near blood vessels where MSC could be found. Through in vitro assays, we confirmed that WT-MSC-conditioned medium was more potent at inducing M2 macrophage polarization, compared with KO-MSC-conditioned medium. VEGF and ET-1 expression in KO-MSC was significantly lower than in WT-MSC, correlating in vivo with reduced tumor growth and numbers of pericytes and M2 macrophages within tumors. Overall, our results suggested that MFG-E8 acts at two levels, by increasing VEGF and ET-1 expression in MSC and by enhancing M2 polarization of macrophages, to increase tumor angiogenesis. Cancer Res; 76(14); 4283-92. ©2016 AACR.