Yong Soo Park;Yeonji Kim;Sung Won Kim; In-Beom Kim
Applied Microscopy
/
v.50
/
pp.11.1-11.3
/
2020
The human turbinate-derived mesenchymal stem cells (hTMSCs), which were DiI-labeled and transplanted into the subretinal space in degenerating mouse retina, were observed in retinal vertical sections processed for rhodopsin (a marker for rod photoreceptor) by confocal microscope with differential interference contrast (DIC) filters. The images clearly demonstrated that DiI-labeled hTMSCs have rhodopsin-immunoreactive appendages, indicating differentiation of transplanted hTMSC into rod photoreceptor. Conclusively, the finding suggests therapeutic potential of hTMSCs in retinal degeneration.
There is increasing evidence that stromal reaction in cancer has an important diagnostic and prognostic significance. The aim of our study is to analyze the relation between the increase in mast cell number and the expression CD34 and alpha-smooth muscle actin (${\alpha}$-SMA) in the stroma of cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). We investigated a total of 29 CIN (1,2,3) and 21 SCC (microinvasive and invasive) specimens and compared the distribution of $CD34^+$ stromal cells, ${\alpha}-SMA^+$ cells, transforming growth factor-${\beta}1$$(TGF-{\beta}1)^+$ cells, and the density of mast cells using immunohistochemistry with antibodies against CD34, ${\alpha}$-SMA, TGF-${\beta}1$, and c-Kit (CD117) respectively. Computerized image analysis was to evaluate the positive area (%) and density of the respective immunoreactive cells. In CIN $CD34^+$ cells were abundant in the stroma but no ${\alpha}-SMA^+$ cells were identified except the wall of blood vessels. $CD34^+$ cells were progressively decreased along the continuum from CIN 2 to microinvasive SCC and not observed in the stroma of invasive SCC. Whereas ${\alpha}-SMA^+$ cells were only observed in the stroma of microinvasive and invasive SCC. We found more intense TGF-${\beta}1$ expression in the increased mast cells in the stroma of invasive SCCs than that in the stroma of CIN. These results indicate that disappearance of $CD34^+$ stromal cells and appearance of ${\alpha}-SMA^+$ cells are associated with the stromal change of CIN to SCC and the transformation of $CD34^+$ stromal cells into ${\alpha}-SMA^+$ cells is mediated by TGF-${\beta}1$ secretions in the stromal mast cell of SCC.
This study attempts to investigate several enteroendocrine cells in the gastrointestinal epithelia of the Korean snakes (Dinodon rufozonatum rufozonotum Rhabdophis tigrina tigrina, Enhydris rufodorsata, Agkistrodon blomhoffii brevicaudus, Agkistrodon saxatilis, Agkistodon calginosus). For a light-microscopical examination of immunocytochemistry, the paraffin sections (5 $\mu$ m) of tissue specimens taken from the various parts of the gastrointestinal tract were stained immunocytochemically by PAP procedure with 10 antisera. The frequency of enteroendocrine cells per unit area (mm$^2$) of each mucosa were counted and the shapes of the cells were observed. In Dinodon rufozonatum rufozonatum, Rhobdophis tigrina tigrina, Enhydris rufodorsata, Agkistrodon saxatilis and Agkistrodon caliginosus, cholecystokinin (CCK)-8, gastrin, pancreatic polypeptide (PP) and serotonin cells were observed. But the freuqency of these immunoreactive cells differ trom each portion of gastrointestinal tracts of all species, respectively. In Agkistrodon blomhoffii brevicaudus, CCK-8, gastrin and serotonin cells were observed. CCK-8 and serotonin cells were found in whole gastrointestinal tracts and gastrin cells were observed in pylorus and mucosa of small intestine. The frequency of these cells was different from each portion. The shapes of CCK-8, gastrin, PP and serotonin cells were pyramidal or oval and closed type in stomach. A large number of these cells were spindle in shape and open type in small intestine and anterior pant of large intestine, whereas some cells were closed type. In posterior part of large intestine and rectum, these cells were oval in shape and closed type.
Journal of Physiology & Pathology in Korean Medicine
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v.26
no.2
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pp.166-174
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2012
To determine the effects of Mahaengeuigam-tang(MHEGT) on obesity, the obesity-related factors (gastrin, CGRP, ghrelin, glucagon-like peptide-1, insulin, orexin, leptin, serotonin, NPY) were investigated in the stomach, pancreas, brain of mice by immunohistochemical methods for 4 weeks after Mahaengeuigam-tang(MHEGT) administration. The change of boy weight decreased in MHEGT administered group than that of control group. The immunohistochemical density of the gastrin and CGRP positive cells on pylorus of stomach increased in MHEGT administered group than that of control group. The number of ghrelin immunoreactive cells on stomach decreased in MHEGT administered groups than that of control group. The immunohistochemical density of GLP-1 in the pancreas decreased in MHEGT administered group than that of control group. The immunohistochemical density of insulin positive cells in the pancreas decreased in MHEGT administered group than that of control group. The immunohistochemical density of orexin and NPY positive neurons in the diencephalon was slightly stronger in MHEGT administered group than that of control group. The immunohistochemical density of serotonin and leptin positive neurons was stronger in MHEGT administered group than that of control group. These results demonstrate that Mahaengeuigam-tang(MHEGT) increased the immunohistochemical density of factors related to appetite inhibitors, and decreased the immunohistochemical density of factors related to stimulator of food intake in stomach, pancreas and brain.
In order to develop procedures for the rapid isolation of recombinant sugar transporter in functional form from away from the endogenous insect cell transporter, gene fusion techniques were exploited. Briefly, BamH1-digested human HepG2 type glucose transport protein cDNA was first cloned into a transfer vector pBlueBacHis, containing a tract of six histidine residues. Recombinant baculoviruses including the human cDNA were then generated by allelic exchange following transfection of insect cells with wild-type BaculoGold virus DNA and the recombinant transfer vector. Plaque assay was then performed to obtain and purify recombinant viruses expressing the human transport protein. All the cell samples that had been infected with viruses from the several blue plaques exhibited a positive reaction in the immnuassay, demonstrating expression of the glucose transport protein. In contrast, no color development in the immunoassay was observed for cells infected with the wild-type virus or no virus. Immunoblot analysis showed that a major immunoreactive band of apparent Mr 43,000~44,000 was evident in the lysate from cells infected with the recombinant baculovirus. Following expression of the recombinant fusion protein with the metal-binding domain and enterokinase cleavage site, the fusion protein was recovered by competition with imidizole using immobilized metal charged resin. The leader peptide was then removed from the fusion protein by cleavage with porcine enterokinase. Final separation of the recombinant protein of the interest was achieved by passage over $Ni^{2+}$-charged resin under binding conditions. The expressed transport protein bound cytochalasin B and demonstrated a functional similarity to its human counterpart.
Kang Gyung Rae;Yuk Sang Won;Ko Hyoung Gon;Lee Kwang Gyu;Lee Chang Hyun
Journal of Physiology & Pathology in Korean Medicine
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v.16
no.6
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pp.1170-1176
/
2002
We examined the effects of Whadamcheongwha-tang(WDCWT) extract on the acetic-acid induced antigastric ulcer in rats. These experiments investigated the numerical changes of gastrin and histamine secreting cells of the gastric mucosa by immunohistochemical method, and the changes of mucin of gastric mucosa by PAS-AB stain methods after the oral administration of WDCWT extract(1.0ml/day) and omeprazole(0.2mg/day) for 1, 3 and 6 weeks. The result are as follows; 1. When WDCWT extract was administrated for 1, 3, 6 weeks, in result, gastrin secreting cells in gastric mucosa were increased compared to the control group. 2. When WDCWT extract was administrated for 1, 3, 6 weeks, in result, the density of immunoreactive gastrin cells was increased compare to the control group. 3. When WDCWT extract was administrated for 1, 3, 6 weeks, in results, the changs of mucosal thickness stained by PAS/PAS-AB was increased compared to the control group. 4. When WDCWT extract was administrated for 1, 3, 6 weeks, in results, the density of PAS stain was decreased compare to control group, but density of AB stain was increased compare to control group. The results suggest that WDCWT extract inhibits a gastric acid secretion in rat gastric mucosa, and is useful in the treatment of the hyperacidity and gastric ulcer.
Objective : Heat shock protein family is related to protective mechanism of cells by environmental changes. This study was performed to evaluate the effect of cryopreservation on the heat shock protein 90 (Hsp90) expression in mouse ovarian tissue. Methods : Cryopreservation of mouse ovarian tissue was carried out by slow freezing method. The mRNA level of Hsp90 expression in both fresh and cryopreserved mouse ovarian tissue was analyzed by RT-PCR. The protein expression of Hsp90 was evaluated by Western blot analysis and immunohistochemistry. Results: The mRNA and protein of Hsp90 were expressed in both fresh and cryopreserved mouse ovarian tissue. The amount of Hsp90 mRNA was increased in cryopreserved ovarian tissue after 60 and 90 minutes after thawing and incubation. The amount of Hsp90 protein was increased in the cryopreserved ovarian tissue after 6 hours of the incubation in Western blot analysis. In immunohistochemical study, Hsp90 protein was localized in cytoplasm of oocytes and granulosa cells. Significant level of immunoreactive Hsp90 protein was detected in theca cells contrast to the weak expression in ovarian epithelial cells. Conclusion: This results showed the increase of Hsp90 expression in both mRNA and protein level in the cryopreserved mouse ovarian tissue. It can be suggested that Hsp90 may play a role in the protective or recovery mechanism against the cell damage during cryopreservaion.
Kim, Ki Yup;Yang, Won Yong;Kwon, Seok Min;Kang, Sang Yoon
Archives of Plastic Surgery
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v.36
no.5
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pp.663-666
/
2009
Purpose: Primary malignant lymphomas of the salivary glands are uncommon. The parotid gland was most frequently involved, followed by the submandibular gland, minor salivary gland and sublingual gland. The most common subtype is mucosa - associated lymphoid tissue(MALT) lymphoma. We experienced a case of salivary MALT lymphoma involving parotid gland duct, so report a case with a review of the literature. Methods: A 65 year old female presented with a palpable mass on the left side of her cheek. There was no clinical or laboratory evidence of pre - existing autoimmune disease. Preoperative facial and neck CT with contrast showed $2.1{\times}1.7cm$ sized, ill defined, homogeneous low density mass near left masseter muscle, and no evidence of other enlarged lymph nodes. Results: At operation, a yellowish oval shaped mass was found slightly adhered to middle portion of the parotid gland duct, meaduring $2{\times}1.5{\times}0.7cm$. Microscopic finding showed that centrocyte - like cells, monocyte B cells and plasma cells were diffusely infiltrated. Immunophenotyping was preformed on fixed section. The majority of the small cells were immunoreactive for the B cell marker CD20. Based on the typical histological findings supported by immunostaining, the mass was defined as MALT lymphoma. Conclusion: We report that very rare case of MALT lymphoma involving parotid gland duct in 65 year old female patient was experienced with clinical characteristics, histologic features and references.
The regional distributions and relative frequencies of some gastrointestinal endocrine cells in the 8 portions (fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) of the gastrointestinal tract of ICR mouse (ICR) with immunohistochemical method using 7 types of specific antisera against somatostatin, serotonin, glucagon, cholecystokinin (CCK)-8, secretin, pancreatic polypeptide (PP) and gastrin. In this study, somatostatin-, serotonin-, glucagon-, CCK-8-, secretin- and gastrin-immunoreactive (IR) cells were identified. Most of these IR cells in the intestinal portion were generally spherical or spindle in shape (open-typed cell) while cells showing round in shape (close-typed cell) were found in the stomach regions occasionally. Their relative frequencies were varied according to each portion of gastrointestinal tract. Somatostatin-IR cells were demonstrated throughout whole gastrointestinal tract except for large intestine. Serotonin-IR cells were detected throughout whole gastrointestinal tract and they were most predominant endocrine cell types in this species of mouse. Glucagon-IR cells were restricted to the fundus and rectum with moderate and a few frequencies, respectively. CCK-8-IR cells were observed in the pylorus, duodenum and ileum with numerous, moderate and rare frequencies, respectively. Secretin-IR cells were restricted to the duodenum and ileum with a few and rare frequencies, respectively. Gastrin-IR cells were restricted to the pylorus with numerous frequency. However, no PP-IR cells were found in this study. In conclusion, some peculiar distributional patterns of gastrointestinal endocrine cells were found in the ICR mouse compared to those of other mammals.
One of the most extensively studied populations of multipotent adult stem cells are mesenchymal stem cells (MSCs). MSCs derived from the human umbilical cord vein (HUC-MSCs) are morphologically and immunophenotypically similar to MSCs isolated from bone marrow. HUC-MSCs are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. Since neural tissue has limited intrinsic capacity of repair after injury, the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesenchymal-like stem cells from the human umbilical cord vein, and studied transdifferentiation-promoting conditions in neural cells. Dopaminergic neuronal differentiation of HUC-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulfoxide (DMSO) and butylated hydroxyanisole (BHA) in N2 medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. HUC-MSCs treated with bFGF, SHH and FGF8 were differentiated into dopaminergic neurons that were immunopositive for tyrosine hydroxylase (TH) antibody. HUC-MSCs treated with DMSO and BHA rapidly showed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including NeuroD1, $\beta$-tubulin III, GFAP and nestin was markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after neural differentiation, we confirmed the differentiation of dopaminergic neurons by TH/$\beta$-tubulin III positive cells. In conclusion, HUC-MSCs can be differentiated into dopaminergic neurons and these findings suggest that HUC-MSCs are alternative cell source of therapeutic treatment for neurodegenerative diseases.
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