Objective: To investigate the clinicopathological characteristics of reactive epithelial hyperplasia and dysplasia in the stomach, as well as the clinical value of mucin special staining and proliferating cell nuclear antigen (Ki-67) in distinguishing the two gastric lesions. Methods: The clinical pathological data of 63 patients with gastric reactive epithelial hyperplasia, 54 patients with low-grade dysplasia, and 63 patients with high-grade dysplasia diagnosed from May 2018 to May 2021 in Beijing Chaoyang Hospital, Capital Medical University, Beijing, China were analyzed. Alcian blue periodic acid Schiff (AB-PAS) and Ki-67 staining were performed to examine the mucin staining pattern, number of Ki-67 positive cells, Ki-67 staining patterns in the three groups of lesions, and histopathologic characteristics. Results: The positive rates of AB-PAS in the reactive epithelial hyperplasia and gastric dysplasia groups were 87.3%(55/63) and 10.3%(12/117), respectively. The expression of AB-PAS in the reactive epithelial hyperplasia was gradually increased from the base to the surface of the epithelium. In low-grade dysplasia and high-grade dysplasia, there was no mucin present in the dysplasia epithelium. The difference between the two groups was statistically significant (P<0.01). The positive rate of Ki-67 in the epithelial reactive hyperplasia (>10%) was 81.0% (51/63), and the positive cells were mainly located in the neck and middle parts of the mucosal glands (58/63, 92.1%). In the low-grade dysplasia group, the positive rate of Ki-67 (>10%) was 90.7%(49/54); the positive cells were mainly located in the upper mucosa (33/54, 61.1%), showing a banded distribution pattern; in the high-grade dysplasia group, the positive rate (>10%) was 95.2%(60/63), and the positive cells were mainly located in the whole mucosa (49/63, 77.8%), showing a diffuse/diffuse scattered distribution pattern. The three groups had statistically different rates and distribution patterns of Ki-67 expression (P<0.01). Conclusion: The gastric epithelial reactive hyperplasia and dysplasia can be differentiated using clinicopathological features, AB-PAS staining and Ki-67 expression pattern.
Alcian Blue ; Humans ; Hyperplasia ; Ki-67 Antigen/metabolism* ; Periodic Acid ; Staining and Labeling ; Stomach Neoplasms/diagnosis*

BACKGROUND: This study aimed to observe the cartilaginous matrix production in SRY (sex determining region Y)-box 9 (SOX9)- and/or telomerase reverse transcriptase (TERT)-transfected chondrocytes from monolayer to three-dimensional (3D) culture.
Alcian Blue ; Cartilage ; Chondrocytes ; Clothing ; Coloring Agents ; Eosine Yellowish-(YS) ; In Vitro Techniques ; Proteoglycans ; Real-Time Polymerase Chain Reaction ; Regeneration ; Telomerase ; Tissue Engineering ; Transfection

BACKGROUND: Articular cartilage damage is still a troublesome problem. Hence, several researches have been performed for cartilage repair. The aim of this study was to evaluate the chondrogenicity of demineralized bone matrix (DBM) scaffolds under cyclic hydrostatic pressure (CHP) in vitro. METHODS: In this study, CHP was applied to human bone marrow mesenchymal stem cells (hBMSCs) seeded on DBM scaffolds at a pressure of 5 MPa with a frequency of 0.5 Hz and 4 h per day for 1 week. Changes in chondrogenic and osteogenic gene expressions were analyzed by quantifying mRNA signal level of Sox9, collagen type I, collagen type II, aggrecan (ACAN), Osteocalcin, and Runx2. Histological analysis was carried out by hematoxylin and eosin, and Alcian blue staining. Moreover, DMMB and immunofluorescence staining were used for glycosaminoglycan (GAG) and collagen type II detection, respectively. RESULTS: Real-time PCR demonstrated that applying CHP to hBMSCs in DBM scaffolds increased mRNA levels by 1.3-fold, 1.2-fold, and 1.7-fold (p < 0.005) for Sox9, Col2, and ACAN, respectively by day 21, whereas it decreased mRNA levels by 0.7-fold and 0.8-fold (p < 0.05) for Runx2 and osteocalcin, respectively. Additionally, in the presence of TGF-β1 growth factor (10 ng/ml), CHP further increased mRNA levels for the mentioned genes (Sox9, Col2, and ACAN) by 1.4-fold, 1.3-fold and 2.5-fold (p < 0.005), respectively. Furthermore, in histological assessment, it was observed that the extracellular matrix contained GAG and type II collagen in scaffolds under CHP and CHP with TGF-β1, respectively. CONCLUSION: The osteo-inductive DBM scaffolds showed chondrogenic characteristics under hydrostatic pressure. Our study can be a fundamental study for the use of DBM in articular cartilage defects in vivo and lead to production of novel scaffolds with two different characteristics to regenerate both bone and cartilage simultaneously.
Aggrecans ; Alcian Blue ; Bone Marrow ; Bone Matrix ; Cartilage ; Cartilage, Articular ; Collagen Type I ; Collagen Type II ; Eosine Yellowish-(YS) ; Extracellular Matrix ; Fluorescent Antibody Technique ; Gene Expression ; Hematoxylin ; Humans ; Hydrostatic Pressure ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Osteocalcin ; Real-Time Polymerase Chain Reaction ; RNA, Messenger

BACKGROUND: Tissue engineering is a multidisciplinary field which attracted much attention in recent years. One of the most important issue in tissue engineering is how to obtain high cell numbers and tissue regeneration while maintaining appropriate cellular characteristics in vitro for restoring damaged or dysfunctional body tissues and organs. These demands can be achieved by the use of three dimensional (3D) dynamic cultures of cells combined with cell-adhesive micro-carriers. METHODS: In this study, human mesenchymal stem cells (hMSCs) were cultured in a wave-bioreactor system for up to 100 days, after seeding on Cultisphere-S porous gelatin micro-carriers. Cell counting was performed at the time points of 7, 12, 17, 31 days and compared to those of hMSCs cultured under static condition. Higher growth and proliferation rates was achieved in wave-type dynamic culture, when cell culture continued to day 31. A scanning electron microscope (SEM) photographs, both live and dead and MTT assays were taken to confirm the survival and distribution of cells on porous gelatin micro-carrier surfaces. The results of histological stains such as hematoxylin and eosin, Masson’s trichrome, Alcian blue and Alizarin red S also showed improved proliferation and tissue regeneration of hMSCs on porous gelatin micro-carriers. CONCLUSION: The experimental results demonstrated the effect and importance of both micro-carriers and bioreactor in hMSC expansion on cell proliferation and migration as well as extracellular matrix formation on the superficial and pore surfaces of the porous gelatin micro-carriers, and then their inter-connections, leading to tissue regeneration.
Alcian Blue ; Bioreactors ; Cell Count ; Cell Culture Techniques ; Cell Proliferation ; Coloring Agents ; Eosine Yellowish-(YS) ; Extracellular Matrix ; Gelatin ; Hematoxylin ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Regeneration ; Tissue Engineering

BACKGROUND/OBJECTIVES: Osteoarthritis (OA) is a major public health issue in Japan and other countries, and foods that prevent or treat OA are in strong demand. Proteins and peptides in chicken meat and bones are known for being rich in functional and nutritional ingredients for the improvement of osteoporosis. We speculated that chicken legs, a food consumed in many regions of the world, may also contain such ingredients. In this study, we aim to (i) evaluate the effect of chicken leg extract (CLE) on the promotion of cartilage matrix production and (ii) identify the active ingredient in CLE that contributes to this function. MATERIALS/METHODS: Artificial CLE digest was prepared, and the acid mucopolysaccharide production-promoting activity of the CLE digest was evaluated by alcian blue staining of ATDC5 cells. CLE was orally administered to rabbits with burr holes in the knee joint of the femur, and the degree of regeneration of cartilage matrix was evaluated. Furthermore, we investigated orally administered CLE-derived peptides in human plasma using LC-MS. From measuring the acid mucopolysaccharide production-promotion activity of these peptides, a molecule considered to be an active ingredient in the CLE digest was identified. RESULTS: CLE digest promoted acid mucopolysaccharide production and facilitated regeneration of cartilage matrix in in vitro and in vivo experiments. Four peptides including phenylalanyl-hydroxyproline (Phe-Hyp) were detected as CLE-derived peptides in human plasma. The effect of CLE was inferred to be due to Phe-Hyp, which was confirmed to be present in the CLE digest. CONCLUSIONS: It was shown that CLE stimulated the production of articular cartilage matrix both in vitro and in vivo, and that CLE could be an effective food for preventing or treating OA. Furthermore, only Phe-Hyp was confirmed as the active compound in the CLE digest, suggesting that the activity of CLE was due to Phe-Hyp.
Alcian Blue ; Cartilage ; Cartilage, Articular ; Chickens ; Femur ; Humans ; In Vitro Techniques ; Japan ; Knee Joint ; Leg ; Meat ; Osteoarthritis ; Osteoporosis ; Peptides ; Plasma ; Public Health ; Rabbits ; Regeneration

Ovarian tumors are relatively uncommon in paediatric age group and mostly occur in the period after menarche and are extremely rare prior to menarche. However, in children and adolescent, the epithelial ovarian tumors account approximately 10% to 28% of all ovarian tumors. In the present case, the patient was presented with abdominal pain for 1 day and no mass was felt on physical examination. Contrast-enhance computed tomography abdomen and pelvis showed a left ovarian cystic mass measuring 7.3×5.1 cm in size and unilateral oophorectomy was done. Tumor shows multiloculated cysts of varying sizes filled with mucinous fluid. Cysts were lined by tall columnar epithelial cells and show predominantly periodic acid-Schiff positive neutral mucin favouring benign nature of lesion. We present this case due to its uncommon age of presentation and the patient was premenarchal girl.
Abdomen ; Abdominal Pain ; Adolescent ; Alcian Blue ; Child ; Epithelial Cells ; Female* ; Fertility ; Humans ; Menarche ; Mucins ; Neoplasms, Glandular and Epithelial ; Ovarian Cysts ; Ovarian Neoplasms ; Ovariectomy ; Pelvis ; Periodic Acid-Schiff Reaction ; Physical Examination

To develop decellularized heart valve scaffold from porcine for heart valve regeneration. Porcine heart valves were decellularized with unique optimized approach by using 1% sodium dodecyl sulfate solution and 5% dimethyl sulfoxide for the first time. Effect of decellularization process on scaffold were characterized by hematoxylin-eosin, 4′,6-diamidino-2-phenylindole, Masson's trichrome, alcian blue staining and scanning electron microscopy for extracellular matrix (ECM) analysis in scaffold. The results showed that developed protocol for decellularization of heart valve scaffold shown complete removal of all cellular components, without changing the properties of ECM. The developed protocol was successfully used for heart valve ECM scaffolds development from porcine. The developed protocol seems to be promising solution for the heart valve tissue engineering application.
Alcian Blue ; Detergents* ; Dimethyl Sulfoxide ; Extracellular Matrix ; Heart Valves* ; Heart* ; Microscopy, Electron, Scanning ; Regeneration ; Sodium Dodecyl Sulfate ; Tissue Engineering*

We report the first published case of multiple eruptive myxoid dermatofibroma (MEMDF) in a male with chronic hepatitis B infection presenting with eruptive lesions showing marked deposits of dermal mucin. Alcian blue and immunohistochemistry confirmed the diagnosis of myxoid dermatofibroma. Further work-up showed asymptomatic chronic hepatitis B infection without cirrhosis.This case highlights an extremely rare histologic variant and the importance of screening for altered immunity in patients with eruptive dermatofibromas.

Human ; Female ; Middle Aged ; Histiocytoma, Benign Fibrous ; Mucins ; Alcian Blue ; Hepatitis B, Chronic ; Immunohistochemistry ; Muc1 Protein, Human ; Mucin-1 ; Liver Cirrhosis ; Exanthema

BACKGROUND/OBJECTIVES: Constipation is a condition that can result from intestinal deformation. Because humans have an upright posture, the effects of gravity can cause this shape deformation. Oligosaccharides are common prebiotics and their effects on bowel health are well known. However, studies of the physiological functionality of a product that contains both lactulose and galactooligosaccharides are insufficient. We investigated the constipation reduction effect of a dual-type oligosaccharide, Dual-Oligo, in loperamide-treated rats. MATERIALS/METHODS: Dual-Oligo consists of galactooligosaccharides (15.80%) and lactulose (51.67%). Animals were randomly divided into four groups, the normal group (normal), control group (control), low concentration of Dual-Oligo (LDO) group, and high concentration of Dual-Oligo (HDO) group. After 7 days of oral administration, fecal pellet amount, fecal weight, water content of fecal were measured. Blood chemistry, short-chain fatty acid (SCFA), gastrointestinal transit ratio and length and intestinal mucosa were analyzed. RESULTS: Dual-Oligo increased the fecal weight, and water content of feces in rats with loperamide-induced constipation. Gastrointestinal transit ratio and length and area of intestinal mucosa significantly increased after treatment with Dual-Oligo in loperamide-induced rats. A high concentration of Dual-Oligo tended to produce more acetic acid than that observed for the control group, and Dual-Oligo affected the production of total SCFA. Bifidobacteria concentration of cecal contents in the high-concentration oligosaccharide (HDO) and low-concentration oligosaccharide (LDO) groups was similar to the result of the normal group. CONCLUSIONS: These results showed that Dual-Oligo is a functional material that is derived from a natural food product and is effective in ameliorating constipation.
Acetic Acid ; Administration, Oral ; Alcian Blue ; Animals ; Chemistry ; Constipation* ; Feces ; Gastrointestinal Transit ; Gravitation ; Humans ; Intestinal Mucosa ; Lactulose ; Loperamide ; Oligosaccharides* ; Posture ; Prebiotics ; Rats* ; Water

Endochondral bone formation is the process by which mesenchymal cells condense into chondrocytes, which are ultimately responsible for new bone formation. The processes of chondrogenic differentiation and hypertrophy are critical for bone formation and are therefore highly regulated. The present study was designed to investigate the effect of aloe-emodin on chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Aloe-emodin treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. ATDC5 cells were treated with aloe-emodin and stained with alcian blue. Compared with the control cells, the ATDC5 cells showed more intense alcian blue staining. This finding suggested that aloe-emodin induced the synthesis of matrix proteoglycans and increased the activity of alkaline phosphatase. Aloe-emodin also enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, BSP and RunX2 in a time-dependent manner. Furthermore, examination of the MAPK signaling pathway showed that aloe-emodin increased the activation of extracellular signal-regulated kinase (ERK), but had no effect on p38 and c-jun N-terminal kinase (JNK). Aloe-emodin also enhanced the protein expression of BMP-2 in a time-dependent manner. Thus, these results showed that aloe-emodin exhibited chodromodulating effects via the BMP-2 or ERK signaling pathway. Aloe-emodin may have potential future applications for the treatment of growth disorders.
Alcian Blue ; Alkaline Phosphatase ; Animals ; Cartilage ; Chondrocytes ; Chondrogenesis ; Collagen ; Growth Disorders ; Hypertrophy ; JNK Mitogen-Activated Protein Kinases ; Mice ; Osteogenesis ; Phosphotransferases* ; Proteoglycans