Scarlet Staining Research Articles

Clinical significance of direct fast scarlet staining on the diagnosis of eosinophilic colitis: A comparative study focusing on the eosinophil degranulation in colonic mucosal tissue.

This study aimed to validate the DFS (direct fast scarlet) staining in the diagnosis of EC (eosinophilic colitis). The study included 50 patients with EC and 60 with control colons. Among the 60 control samples, 39 and 21 were collected from the ascending and descending colons, respectively. We compared the median number of eosinophils and frequency of eosinophil degranulation by HE (hematoxylin and eosin) and DFS staining between the EC and control groups. In the right hemi-colon, eosinophil count by HE was useful in distinguishing between EC and control (41.5 vs. 26.0 cells/HPF, p < 0.001), but the ideal cutoff value is 27.5 cells/HPF (high-power field). However, this method is not useful in the left hemi-colon (12.5 vs. 13.0 cells/HPF, p = 0.990). The presence of degranulation by DFS allows us to distinguish between the groups even in the left hemi-colon (58% vs. 5%, p < 0.001). DFS staining also enabled a more accurate determination of degranulation than HE. According to the current standard to diagnose EC (count by HE staining ≥20 cells/HPF), mucosal sampling from left hemi-colon is problematic since the number of eosinophils could not be increased even in EC. Determination of degranulated eosinophils by DFS may potentiate the diagnostic performance even in such conditions.

SphK1 deficiency ameliorates the development of atherosclerosis by inhibiting the S1P/S1PR3/Rhoa/ROCK pathway

Background and aimsS1P is an important factor regulating the function of the vascular endothelial barrier. SphK1 is an important limiting enzyme for the synthesis of S1P. However, the role of the SphK1/S1P-mediated vascular endothelial barrier function in atherosclerosis has not been fully revealed. This study explored the roles and mechanisms of SphK1 on atherosclerosis in vivo and in vitro. MethodsIn vivo, ApoE−/− and SphK1−/−ApoE−/− mice were fed a high-fat diet to induce atherosclerosis. In vitro, ox-LDL induced HUVECs to establish a cell model. Aortic histological changes were measured by H&E staining, Oil Red O staining, EVG staining, Sirius scarlet staining, immunofluorescence, and Evans Blue Assay. Western blotting was performed to explore the specific mechanism. ResultsWe validated that deficiency of SphK1 resulted in a marked amelioration of atherosclerosis, as indicated by the decreased lipid accumulation, inflammatory factors, oxidative stress, aortic plaque area, inflammatory factor infiltration, VCAM-1 expression, and vascular endothelial permeability. Moreover, deficiency of SphK1 downregulated the expression of aortic S1PR3, Rhoa, ROCK, and F-actin. The results of administration with the SphK1 inhibitor PF-543 and the S1PR3 inhibitor VPC23019 in vitro further confirmed the conclusion that deficiency of SphK1 reduced S1P level and S1PR3 protein expression, inhibited Rhoa/ROCK signaling pathway, regulated protein expression of F-actin, improved vascular endothelial dysfunction and permeability, and exerted anti-atherosclerotic effects. ConclusionsThis study revealed that deficiency of SphK1 relieved vascular endothelial barrier function in atherosclerosis mice via SphK1/S1P/S1PR signaling pathway.

Four-dimensional computed tomography is useful for detection of apical sparing of cardiac amyloidosis.

A 73-year-old male was admitted because of recurrent syncope. He was diagnosed with transient bradycardia caused by a 2:1 atrioventricular block, and he underwent cardiac computed tomography (CT) using 320 detector-row CT to screen for coronary artery disease. Significant coronary artery stenosis was not detected, but diffuse late iodinate enhancement was found on the epi-myocardium and endo-myocardium of the interventricular septum, and endo-myocardium of the anterior and lateral left ventricular (LV) myocardium (LVM) on CT. The ejection fraction and global longitudinal strain (LS) of LVM were 53.97% and - 9.87% on CT. Apical sparing was present, meaning the LS of LV apical segments were preserved compared with basal segments on CT. Pathological findings of LVM demonstrated loss of myocardial cells and extra-cellular amyloid deposition on the direct fast scarlet staining. He was finally diagnosed with transthyretin amyloidosis.

Establishment and refinement of a DEN-induced hepatocellular carcinoma model in rats

Abstract Objectives Hepatocellular carcinoma is one of the most common malignant tumors in the world with complex etiology, high tumor heterogeneity, and low efficacy of treatment. The establishment of an animal model that is close to the clinical situation of hepatocellular carcinoma and can be successfully modeled many times is of great significance to the study of the pathogenesis, diagnosis and treatment of hepatocellular carcinoma. Methods We used Diethylnitrosamine (DEN) to induce hepatocellular carcinoma in rodents and compared four models of DEN-induced hepatocellular carcinoma. Group C (Control): rats were fed a standard laboratory rat diet and freely drank normal water. Group P (Peritoneal injection): rats were administered an IP injection (50 mg/kg/week) between 5 and 23 weeks after birth. Ten microliter of DEN solution would be injected per g of rat. Group O (DEN-Fed group): rats were allowed unrestricted access to water contaminated with 0.01 % DEN between the ages of 7 and 15 weeks. 0.2 mL of DEN drinking water was consumed per gram of rat. Group P+O (Combined peritoneal injection and Oral feeding): rats were administered an IP injection (50 mg/kg) at weeks 3 and 5 post-birth, and they freely drank water contaminated with 0.012 % DEN between weeks 7 and 14 post-birth. We used an ultrasound scan, biochemical testing, haematoxylin, and eosin staining, Masson staining, Wolf scarlet staining, Ki67, CD34, a-SMA, CD8, and CD68 staining to compare between groups. Results Liver dissection and ultrasound scan showed that compared to other groups, the liver of Group P+O was darker in color, with more grey-white cancer nodules and larger localized tumors, and the structure of the tumors was slightly disorganized, with the elastography hardness of the middle lobe and the right lobe was slightly increased. The alanine aminotransferase and total bilirubin of Group P+O were higher than those of Group O but lower than those of Group P. Haematoxylin and eosin staining showed that the tumors of Group P+O were large, with large tumor cords and pseudo-glandular, the degree of differentiation was medium and surrounded by more fatty lesions. Conclusions We conclude that combined DEN treatment is more effective, stable, and has the advantage of multiple modalities, leading to faster tumor formation.

Discordance of light chain isotypes between serum and glomerular deposits in proliferative glomerulonephritis with monoclonal IgG deposits: a case report and review of the literature

BackgroundProliferative glomerulonephritis with monoclonal immunoglobulin G (IgG) deposits (PGNMID) is a disease entity with nonorganized granular glomerular deposition with monoclonal proteins of both heavy and light chains. Dysproteinemia was observed in only 30% of the patients with PGNMID. We herein report a case of PGNMID with discrepancy between serum and glomerular deposits.Case presentationThe patient was a 50-year-old man who had been followed at a local clinic due to hypertension, type 2 diabetes, hyperlipidemia, hyperuricemia, fatty liver, and obesity. Proteinuria had been noted five years previously, and he had been referred to a hematology department due to hyperproteinemia, high gamma globulin, and κ Bence-Jones protein (BJP) positivity one year previously. Bone marrow aspiration showed 5% plasma cells, and he was referred to the nephrology department to evaluate persistent proteinuria. He was hypertensive, and his estimated glomerular filtration rate was 54.2 ml/min/1.73 m2. His urinary protein level was 0.84 g/g⋅Cr. Urine and serum immunofixation showed BJP-κ type and IgG-κ type, respectively. Kidney biopsy showed an increase in mesangial cells and matrix without nodular lesions under a light microscope. Immunofluorescence microscopy showed granular deposits of IgG and C3 on the capillary wall and weak positivity for C1q. IgG3 was predominant among the IgG subclasses, and intraglomerular κ and λ staining was negative for κ and positive for λ. Direct fast scarlet staining was negative. Electron microscopy showed lumpy deposits without a fibrillar structure in the subepithelial area. Based on the above findings, a diagnosis of membranous nephropathy-type PGNMID was made. Since proteinuria increased gradually after three years of treatment with valsartan (40 mg, daily), oral prednisolone (30 mg, daily) was initiated, which led to decreased proteinuria. The dose of oral prednisolone was gradually tapered to 10 mg per day. At that time, proteinuria was 0.88 g/g⋅Cr. We found 204 cases in 81 articles in the PubMed database, among which 8 showed discrepancy in the heavy and/or light chains between serum and kidney.ConclusionsWe experienced a case of membranous nephropathy-type PGNMID with discrepancy in light chains between serum and kidney that was successfully treated with oral prednisolone.

Colonic amyloidosis: Rare endoscopic findings mimicking a depressed neoplastic lesion.

A colonic depressed lesion was found in a 77-year-old man and suspected as a neoplastic lesion using conventional endoscopy. Magnifying endoscopy, however, did not support it and biopsy revealed the lesion to be AL amyloidosis.

Experimental Study of Using Aloe Vera Gel to Treat Diabetic Chronic Cutaneous Ulcers in Bama Miniature Pigs

To investigate the efficacy, safety, and mechanism of topical application of aloe vera gel (AVG) to treat diabetic chronic cutaneous ulcers in Bama miniature pigs. The Bama miniature pig model of diabetic chronic skin wounds was constructed and the model pigs were randomly assigned to AVG daily administration group (AVG QD), aloe vera gel every-other-day administration group (AVG QOD), and diabetic control group (DC). A non-diabetic chronic skin wounds model pig was set as the non-diabetic control group (NDC). Treatment efficacy was evaluated based on the amount of time needed for complete healing of the wounds, healing rates, granulation growth rates, and skin histopathological changes. Safety was evaluated according to whether adverse reactions were observed. In addition, the dynamic changes of the relative expression levels of miR21, miR29a, miR126, miR146a, miR155, and miR210 in wound granulation tissues were examined. 1) Efficacy and safety: The amount of time needed for complete healing of the wounds was shorter in the NDC group than those of the three other groups, DC group, AVG QD group, and AVG QOD group (all P<0.05). The amount of time needed for complete healing of the wounds was shorter in the AVG QD group and AVG QOD group than that of DC group (all P<0.05). The amount of time needed for complete healing of the wounds was shorter in the AVG QOD group than that of AVG QD group (all P<0.05). No adverse reactions were detected in the whole process of AVG topical treatment. The granulation growth rate of NDC group was higher than those of DC group, AVG QD group, and AVG QOD group (all P<0.05). The wound healing rate of NDC group was higher than those of DC group, AVG QD group, and AVG QOD group (all P<0.05); the wound healing rate of AVG QOD group was higher than those of DC group and AVG QD group (all P<0.05). 2) Histopathology: The results of HE staining light microscopy showed that collagen fiber production increased, and that microvascular formation with slight inflammatory cell infiltration was observed in the dermal interstitium at the initial stage of wound healing after AVG treatment. One year of after complete healing, pathological examination results of wound healing skin showed that the epidermal keratinization was complete, that collagen was arranged neatly and orderly, and that many microvessels were found in the interstitium. The results of picric acid celestite scarlet staining showed that, after AVG treatment, type Ⅰ collagen mainly increased in the initial stage of wound healing, type Ⅲ collagen gradually increased when the wound healed completely, and the collagen was arranged neatly during the whole process. 3) The relative expression of microRNAs: The relative expression of miR21, miR126, and miR210 in NDC group, AVG QD group, and AVG QOD group were higher than that in DC group (all P<0.05). The relative expression of miR29a and miR155 in NDC group, AVG QD group, and AVG QOD group was lower than that in DC group (all P<0.05). The relative expression of miR146a in NDC group was higher than that in DC group ( P<0.05). AVG topical application can shorten the time needed for complete healing of diabetic chronic wounds in Bama minipigs. The wound healing speed of the alternate-day treatment group was faster than that of the daily treatment group. No adverse reactions were observed over the course of the treatment. The mechanism may be related to the up-regulation of the expressions of miR21, miR126, and miR210 and the down-regulation of miR29a and miR155 in wound granulation tissue.

Massive eosinophilia despite severe aplastic anemia.

A 47-year-old nonsmoking man developed very severe aplastic anemia (AA) (hemoglobin 56 g/L, neutrophils 110/μl, eosinophils 0/μl, platelets 4000/μl) with fatty marrow (Figure 1A: hematoxylin–eosin stain, ×4 objective). He had a history of bronchial asthma for 3 years and had been treated with inhaled corticosteroid in combination with long-acting β2-agonist (budesonide/formoterol fumarate). In addition, he had developed eosinophilic pneumonia requiring systemic administration of prednisolone (<30 mg/day) twice in the last 1.5 years. His family history was not significant for any serious diseases. Treatment with antithymocyte globulin, eltrombopag olamine, and cyclosporine (CsA) (500 mg/day) eliminated the need for blood transfusions and the CsA dose was gradually reduced. While pancytopenia continued (hemoglobin <97 g/L, neutrophils <1400/μl, platelets <30,000/μl), eosinophilia (>500/μl) appeared 17 months after the start of treatment. This increased to 4550/μl in 4 months after discontinuation of CsA treatment at 33 months. In addition, he presented with a rash with purpura on the legs, for which prednisolone (<20 mg/day) was not effective. Bone marrow examination showed mild hypocellular marrow with eosinophil expansion (43.8%) (Figure 1B: hematoxylin–eosin stain, ×4 objective; Figure 1C: hematoxylin–eosin stain, ×40 objective; Figure 1D: direct Fast Scarlet stain, ×40 objective). Increased blasts and significant dysplasia were not observed. G-banding and fluorescence in situ hybridization for the FIP1L1–PDGFRA re-arrangement detected no chromosomal aberrations. Moreover, skin biopsy revealed eosinophilic vasculitis and panniculitis, compatible with eosinophilic granulomatosis with polyangiitis (Figure 1E: hematoxylin–eosin stain, ×20 objective; Figure 1F: direct Fast Scarlet stain, ×40 objective). After re-administration of CsA (150 mg/day), eosinophilia and skin lesions disappeared in 5 weeks. This case indicates that massive reactive eosinophilia can occur even in AA patients with eosinophil-associated diseases, especially during dose reduction, or after the discontinuation of CsA. None. The authors declare that there is no conflict of interest. Aya Takahashi and Tsutomu Shinohara designed the research, drafted the manuscript, and created the figure. Aya Takahashi performed the skin biopsy. Yoshihito Iwahara, Aya Takahashi, Hisanori Machida, Keishi Naruse, Eiji Takeuchi, and Tsutomu Shinohara analyzed the data. Keishi Naruse contributed to pathological assessment. All authors revised the manuscript critically and approved the final manuscript.

Intestinal flora polymorphisms with different lesional stages in an animal model of MAFLD

Objective: To study the intestinal flora specific differences with different lesional stages of metabolic (disorder) associated fatty liver disease (MAFLD), namely simple steatosis and steatohepatitis, so as to provide a new direction for MAFLD-related intestinal flora transplantation and targeted therapy. Methods: Mice were fed with normal diet, methionine-choline deficient diet (MCD) and a high-fat high-fructose diet (HFHF) for 12 weeks to construct simple steatosis and steatohepatitis models. HE and Sirius scarlet staining was performed to observe the liver pathological changes. The qPCR method was used to evaluate inflammation and liver fibrosis factors. A fully automatic biochemical analyzer was used to detect changes in liver transaminase and blood lipids. 16S rRNA sequencing method was used to observe the intestinal flora differences in the feces of each group of mice. The comparison of means between two groups was performed by t-test, and the comparison of means between multiple groups was performed by one-way analysis of variance. Kruskal-Wallis rank sum test was used for non-normally distributed data. Results: NAFLD scores were determined with pathological sections (HE and Sirius scarlet staining) of mice liver, which showed that the inflammation and liver fibrosis scores of the MCD and HFHF groups were 2.12 ± 0.18 and 1.06 ± 0.24, and 2.22 ± 0.16 and 0.46 ± 0.10, respectively. The degree of liver inflammation and fibrosis was significantly higher in the MCD than the HFHF group (P < 0.001 and P < 0.01). Lipid deposition was higher in the HFHF than the MCD group (P < 0.001), and the scores were 2.36 ± 0.17 and 1.60 ± 0.24 respectively. Simultaneously, the inflammatory [tumor necrosis factor-A (TNF-a), chemokine factor-2 (CXCL-2)] and hepatic fibrosis indicators [vascular smooth muscle actin alpha (a-SMA) and connective tissue growth factor (CTGF)] had confirmed the above-mentioned results at the transcription level. Moreover, the intestinal flora diversity was reduced (P < 0.05) in the MCD group than the HFHF group, and the Simpson and Shannon index were 0.31 ± 0.10 and 0.42 ± 0.05, and 2.03 ± 0.33 and 1.70 ± 0.28, respectively, and the differences were significant between different intestinal flora groups. The levels of Desulfovibrio, Odoribacter, and Roseburia flora were significantly increased in the HFHF than the MCD group, and the levels of Faecalibaculum, Parasutterella, Alipis, Butyricimonas_virosa, Turicibacter_sp, and Romboutsia_ilealis were significantly increased in the MCD than the HFHF group, and the difference was statistically significant (P < 0.05). Conclusion: There are significant differences in intestinal flora diversity between simple steatosis and steatohepatitis models. Therefore, clarifying the difference between the two may provide a new direction for the stage manner treatment of MAFLD.

Effects of two dusting methods on histopathology and cytokines in rat lung tissue

Objective: To investigate the Effects of different dusting methods on pathological changes and cytokines in rat lung tissue. Methods: 84 healthy male SPF Wistar rats, were randomly divided into control group and dynamic dusting group, tracheal perfusion group, On the day 3, 14, 28, and 60 after the dust exposure, 7 rats in each group were randomly selected and killed. Record the weight of rats and calculate the lung coefficient; Observe the pathological changes of lung tissue by HE staining, Evaluate the pulmonary collagen fibrosis by Masson staining, observe The changes of area ratio of collagen fibers in lung tissue Sirius scarlet staining, Use enzyme-linked immunosorbent assay to detect the content of transforming growth factor (TGF) -β1 and hydroxyproline (HYP) in lung tissue homogenate. Results: After the rats were exposed to dust, the dynamic dusting group and the tracheal perfusion group varied significantly in lung histopathology at four time points, and alveolar inflammation and Pulmonary fibrosis scores were higher than the control group (P<0.05) ; The percentage of type I and type III collagen fiber area of tracheal perfusion group was higher than that in the dynamic dusting group in the four time points (P<0.05) ; The content of TGF-β1 in lung tissue homogenate of rats in the tracheal perfusion group was higher than that in the dynamic dusting group on the day 3 and 14 (P<0.05) ; The content of Hyp in the tracheal perfusion group was higher than that in the dynamic dusting group on the day 28 (P<0.05) . Conclusion: The acute lung injury of rats can be caused by dynamic dusting group and tracheal perfusion group. The lung histopathology shows inflammation and fibrosis, but the lung tissue injury of rats is more serious and the pathological changes are more obvious.

Monoclonal gammopathy of renal significance (MGRS)-related AL amyloidosis complicated by amyloid myopathy: a case report

BackgroundLately, monoclonal gammopathy of renal significance (MGRS) has been defined as a group of renal disorders that are strongly associated with monoclonal protein, including amyloid immunoglobulin light chain (AL) amyloidosis. Amyloid myopathy is rare (1.5% of all patients with amyloidosis) and the prognosis is poor. Furthermore, only approximately 20% of patients with amyloid myopathy are reported to have renal involvement, indicating a lack of data in the literature.Case presentationHere, we report a rare case of MGRS-related AL amyloidosis complicated by amyloid myopathy that presented with muscle weakness in the upper and lower limbs, neck and fingers, and nephrotic syndrome. Blood, urine, and bone marrow examination revealed monoclonal gammopathy of undetermined significance (MGUS) (Bence Jones protein-lambda). Muscle biopsy of the vastus lateralis muscle demonstrated amyloid proteins in the sarcolemma and in the blood vessel walls on Congo red staining, suggesting amyloid myopathy, and tiny inclusions in fibers on modified Gomori trichrome stain. Although we thought they were reminiscent of nemaline bodies, we could not confirm the nature of this structure. Renal biopsy demonstrated amyloid proteins in the mesangial region, part of the capillary walls, and the blood vessel walls on direct fast scarlet staining. As these amyloid proteins were positive for p-component staining and negative for amyloid A staining, β2-microglobulin, and pre-albumin, and as lambda light chains were positive in the mesangial region, we diagnosed the patient with MGRS-related AL amyloidosis. Although he was treated with melphalan and dexamethasone, his symptoms did not improve.ConclusionsAL amyloidosis involving the kidneys and muscles has a poor prognosis, and a delayed diagnosis of amyloid myopathy is common because of its rarity and frequent misdiagnosis, which increases organ function deterioration. Therefore, early detection, therapeutic intervention, and careful follow-up are crucial.

Utility of Direct Fast Scarlet Staining in the Histopathological Diagnosis of Eosinophilic Esophagitis: A Short Report

Eosinophilic esophagitis (EoE), an atopic disease of the esophagus, has become increasingly recognized over the last 15 years. The epidemiology of EoE has now been reported from many countries around the world. While the clinical diagnosis of this disease depends on the patient’s clinical manifestations, the final diagnosis should be made based on the histopathological examination of esophageal mucosal biopsies. In the diagnosis of EoE, to facilitate the appropriate treatment of patients, it is extremely important to precisely recognize the presence of eosinophils in biopsy specimens of the esophageal mucosa. If eosinophils are present, EoE patients should be referred to an allergist for appropriate management with dietary modification, pharmacological agents (including corticosteroids), and/or mechanical dilation of the esophagus. We herein present and recommend the use of direct fast scarlet staining for the easy and precise recognition of eosinophils in biopsy specimens of the esophageal mucosa, a technique that has been routinely used in our laboratory.

Xanthoangelol Isolated from Angelica keiskei Roots Prevents Dextran Sulfate Sodium-Treated Colitis in Mice

Background: The therapeutic effects of a number of natural products on Inflammatory Bowel Disease (IBD) have recently been examined in detail. The whole herb and roots of Angelica keiskei (Umblliferae) have traditionally been used as a diuretic, to treat gastrointestinal diseases such as gastric ulcers and diarrhea in Japan. Objectives: The present study was performed to investigate the effects of xanthoangelol, a major chalcone of Angelica keiskei roots, on diarrhea and inflammation in the large intestine of IBD model mice. Methods: Xanthoangelol (10 &amp; 25 mg/kg) was orally administered to mice with 3% Dextran Sulfate Sodium (DSS)-induced colitis. Blood samples were collected during the experimental period, subjected to a full blood count test, and colonic cytokine and chemokine levels were measured. Results: Xanthoangelol (25 mg/kg) reduced the Disease Activity Index (DAI) of colitis. It also attenuated DSS-induced reductions in red blood cell and platelet counts as well as Hb and Ht levels. A histological examination of the colon using direct fast scarlet staining showed that xanthoangelol prevented DSS-induced mucosal ulceration and eosinophil infiltration. Xanthoangelol also reduced DSS-induced increases in colonic MCP-1, IL-1β, and TNF-α levels. Conclusions: Xanthoangelol reduced DSS-induced increases in colonic IL-1β, TNF-α, and MCP-1 levels and prevented eosinophil infiltration, which supports its potential as a treatment for IBD.

IgM monoclonal gammopathy with heavy-and-light-chain amyloidosis resembling fibrillary glomerulonephritis determined by tandem mass spectrometry: a case report

BackgroundFibrillary glomerulonephritis (FGN) is distinguished from amyloidosis by thicker fibrils and the lack of staining with histochemical dyes typically reactive with amyloid. However, congophilic FGN has been proposed recently and adding laser microdissection followed by mass spectrometry (LMD/MS) to conventional pathological methods would be helpful to diagnose FGN. Here, we report a patient initially diagnosed with FGN whose final pathological diagnosis was changed to immunoglobulin heavy-and-light-chain amyloidosis (AHL) after LMD/MS.Case presentationA 75-year-old male developed nephrotic syndrome. Protein electrophoresis showed IgM κ type M proteinemia and he was diagnosed with IgM monoclonal gammopathy. A renal biopsy was performed and pathological examination showed marked periodic acid-Schiff-positive enlargement of the mesangial region and silver stain positivity, but weak direct fast scarlet staining. Immunofluorescence analysis showed monoclonal deposition of IgM-κ chain in the glomerulus. Under electron microscopy, the fibrils were about 20 nm in diameter, which was thicker than typical amyloid fibrils. Based on these findings, the patient was diagnosed with FGN. Although cyclophosphamide and prednisolone were administered, his renal function deteriorated and progressed to end stage renal disease requiring maintenance hemodialysis. As congophilic FGN has been recognized since 2018, Congo red staining and LMD/MS were performed. The Congo red staining was positive and LMD/MS results indicated that this was a case of AHL.ConclusionsWe reported a case of μ and κ chain AHL resembling FGN requiring LMD/MS for definitive diagnosis. Since FGN and amyloidosis exhibit pathological findings, even if Congo red staining is positive, LMD/MS needs to be considered in cases atypical pathological findings, such as silver stain positivity or thicker fibrils.

Preventive Effects of Mallotus japonicus Cortex Extracts on Dextran Sulfate Sodium-Induced Ulcerative Colitis in C57 BL/6J Mice

Background: The cortex of Mallotus japonicus (Euphorbiaceae) has traditionally been used to treat gastric ulcers, duodenal ulcers, and gastric hyperacidity in Japan. A large number of studies have recently focused on its effects on Inflammatory Bowel Disease (IBD). Objective: The aim of the present study was to examine the effects of M. japonicus (MJ) extracts on large intestinal diarrhea and inflammation using Inflammatory Bowel Disease (IBD) mouse models. Methods: The present study used 3% Dextran Sulfate Sodium (DSS)-treated colitis models. Red blood cell, platelet, and leukocyte counts in addition to hematocrit (Ht), hemoglobin (Hb), and colonic cytokine and chemokine levels were measured in DSS-treated C57BL/6J mice during the experimental period. Results: The Disease Activity Index (DAI) was lower in 3% DSS-treated mice orally administered MJ (200 and 500 mg/kg) than in mice administered 3% DSS only. Furthermore, MJ inhibited decreases in red blood cell and platelet counts as well as Hb and Ht levels in DSS-treated mice. Colon histology using direct fast scarlet staining revealed that MJ prevented mucosal membrane ulceration and eosinophil infiltration of the mucosal membrane induced by the DSS treatment. Increases in colonic Monocyte Chemoattractant Protein 1 (MCP)-1, interleukin (IL)-1β, and Tumor Necrosis Factor (TNF)-α levels in DSS-treated mice were reduced by orally administered MJ extracts. Conclusion: The present results suggest that M. japonicus cortex extracts are an effective treatment for IBD through the inhibition of increases in colonic IL-1β, TNF-α, and MCP-1 levels and eosinophil infiltration of the colon in DSS-treated mice.

Effects of kampo medicine keishikashakuyakuto on large intestine disease in mice

ABSTRACTAimThe aim of this study was to examine the effects of keishikashakuyakuto (KST) on large intestine diarrhea and inflammation in irritable bowel syndrome (IBS) and inflammatory bowel disease (IBD) mouse models.MethodsThis study examined the effects of KST on diarrhea and inflammatory bowel syndrome using 5‐hydroxytryptamine (5‐HT)‐induced diarrhea and 3% dextran sulfate sodium (DSS)‐treated colitis models.ResultsKeishikashakuyakuto (200 and/or 500 mg/kg) and ramosetron (5 μg/kg) prolonged the onset‐time of 5‐HT‐induced diarrhea in mice. KST and ramosetron prevented diarrhea and the stool score was decreased 60 min after 5‐HT treatment. The disease activity index in the 3% DSS‐treated mice was lower after oral KST than in mice given 3% DSS only. Furthermore, KST inhibited the decreases in red cells and platelets, and in hemoglobin and hematocrit that were observed in DSS‐treated mice. On colon histology using hematoxylin–eosin and direct fast scarlet staining, KST prevented the mucosa membrane ulceration and eosinophil infiltration of the mucosa membrane caused by DSS treatment. The increase in colonic monocyte chemoattractant protein 1, interleukin‐1β and tumor necrosis factor‐α in DSS‐treated mice was reduced by oral KST.ConclusionKeishikashakuyakuto may be effective for IBS and IBD.

脾破裂と後腹膜血腫を契機に発見されたアミロイドーシスの1例(A case of amyloidosis diagnosed as spontaneous splenic rupture and retroperitoneal hemorrhage)

要旨 我々は，当初は外傷の可能性も考慮されたが，結果的にALアミロイドーシスと診断された体幹部多発出血の1例を経験し，適切な治療にて救命し得たので報告する。症例は高血圧の既往がある74歳の女性で，自宅で動けなくなっているところを発見・救急要請され，前医に搬送された。前医で行った腹部CT検査で後腹膜血腫を認め，骨盤骨折の疑いでドクターヘリが要請された。フライトドクターが現場でFAST（focused assessment with sonography for trauma）陽性と判断した。病院到着後，腹腔内液体貯留が増加，急速輸液に反応しないショック状態に対して，緊急開腹手術を行った。術中所見では，腫大脾の破裂がみられ，脾臓摘出術を行った。また，続いて行った血管造影検査にて，右腰動脈からのextravasationが確認され，コイル塞栓術を行って止血を完了した。摘出した脾臓組織の病理検査を行い，ALアミロイドーシスと診断された。その後，患者は退院し，外来通院中である。アミロイドーシスにおいて腫大脾が破裂・出血する症例はあるが，後腹膜出血を同時に併発した報告は検索する限り皆無であった。本症例のように外傷機転のない体幹多発出血の原因には，アミロイドーシスを鑑別に挙げるべきである。

Unique acidophilic inclusions in the submucosal layer of colon – a case report

We report a case of colon cancer which revealed unique acidophilic inclusions in the submucosal layer of colon broadly. The patient was a 70-year-old male. He underwent surgery for transverse colon cancer. The cancer invaded the subserosal layer. There was a large number of acidophilic inclusions in the submucosal layer of the colon broadly. The inclusions were found around the cancer and were invaded by the cancer. The inclusions had clear borders and included fat cells, vessels, nerve plexus, collagen fibers, and elastic fibers. They were periodic acid-Schiff stain negative and did not show positivity for amyloid in the direct fast scarlet stain. In ultrastructual study, the inclusions revealed homogenous dense material accompanied by collagen fibers and elastic fibers. We cannot find any literature about the unique inclusions, and conclude they are unknown material deposits in the colonic wall.

The characteristics of type I, III collagen and LN in pulmonary fibrosis induced by uranium ore dust in rats

To explore the characteristics of LN and type I, III collagen in pulmonary fibrosis induced by uranium ore dust in rats. 60 adult Wistar rats were divided randomly into two groups, control group (30 rats) and uranium ore dust group (30 rats). Non-exposed intratracheal instillation method was used. Uranium ore dust group was exposed 20 mg/ml uranium ore dust suspension 1ml per rat, meanwhile control group was exposed normal saline 1ml per rat. Post-exposed the 7, 14, 21, 30 and 60 d, 6 rats in each group were killed randomly, lung tissue were collected. The pathological changes in lung tissue were observed by microscope using HE staining, the collagen I and III in lungs were observed by polarizing microscope using Biebrich scarlet staining. The expression of LN protein in lung tissue was observed by immunohistochemistry-SP. During lung fibrosis, a large amount of the proliferated I and III collagen in lungs were observed. Post-exposure to uranium ore dust, the characteristics in proliferated collagen in lungs were type I collagen deposited in lung interstitium mainly in the early stage. The area percentage of collagen I and III was increased significantly at 7, 14, 21, 30 and 60d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01). The over expression of LN in the lung tissue were observed. The expression of LN was distributed in the lung tissue as thickening of the linear or cluster. The integral optical density of LN was increased significantly at 21, 30 and 60 d in the experimental group as compared with that in the control group (P < 0.05 or P < 0.01). After exposure to uranium ore dust, the characteristics in proliferated collagen in lungs are the type of I collagen deposited in lung interstitium mainly in the early stage, while the type of III collagen increase significantly at the later period. The overexpression of LN exists in the process of pulmonary fibrosis. It suggests that LN has a role effect in the process of pulmonary fibrosis.

Correlation of MMP1 and TIMP1 expression with pituitary adenoma fibrosis

To explore the correlation of matrix metalloproteinase-1 (MMP(1)) and tissue inhibitor of metalloproteinase-1 (TIMP(1)) with pituitary adenoma fibrosis. Thirty-eight pituitary adenoma specimens were divided into fibrous group (6 patients) and non-fibrous group (32 patients). MMP(1) and TIMP(1) expression was detected by RT-PCR and immunohistochemistry. The collagen content was determined by Sirius scarlet staining. In fibrous and non-fibrous group: (1) the collagen content was 20.95 +/- 8.42% and 7.98 +/- 5.18% respectively, and there was a statistical significance (P < 0.01). (2) The expression of MMP(1)mRNA was 0.47 +/- 0.40 and 0.59 +/- 0.54 respectively and its protein expression was 0.12 +/- 0.09 and 0.13 +/- 0.09 respectively, and there was no statistical significance (P > 0.05). Their expression was not related to collagen content (P > 0.05). (3) The expression of TIMP(1)mRNA was 1.61 +/- 1.09 and 0.79 +/- 0.59 respectively and its protein expression was 0.58 +/- 0.11 and 0.32 +/- 0.18 respectively, and there was a statistical significance (P < 0.01). Their expression was related to collagen content (P < 0.01). The pathological features of pituitary adenoma fibrosis are excessive collagen deposition. High expression of TIMP(1) may be an important cause.