Enhanced Expression of Carbonic anhydrase II in Hypokalemic Rat Kidney / 체질인류학회지

A number of acid-base or electrolyte disorders are associated with decreased or increased HCO3- reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Carbonic anhydrase II in the kidneys of normal and potassium-depleted rats using Western blot analysis and immuno-histochemistry. Western blot analysis demonstrated that CA II protein, ~30 kDa at molecular mass, was abundantly expressed in normal group. All potassium-depleted groups showed slightly increased CA II protein compared to normal group. In control group, immunoreactivity of CA II protein was detected in the entire collecting duct. Signal intensity was prominent in the intercalated cells and weak in the principal cells of the cortical collecting ducts. In potassium-depleted groups, the pattern of cellular labeling of CA II protein was identical to that of normal group, but the signal intensity was decreased in cortical collecting duct, markedly increased in the inner stripe of outer medullary and inner medullary collecting ducts, and unchanged in the outer stripe of outer medullary collecting duct. These results suggest that chronic hypokalemia impact the expression pattern of CA II protein depending the portion of the collecting duct.

Alterations of Aquaporin-1, 2, 3 Protein Expression in Pregnant Rat Kidney / 대한해부학회지

The pregnancy causes the marked change in maternal renal hemodynamic and volume homeostasis. During pregnancy, renal sodium and water retention result in an expansion of extracellular fluid and plamsma volume. Although many studies suggested that water balance or water balance disorder was associated with regulation of Aquaporin (AQP) expression, the studies were only limited to AQP-2 expression during the pregnancy. The present study was to examine altered expression and distribution of AQP-1, 2, and 3 proteins in the kidneys of non-pregnant (NP) and pregnant rats using Westhern blot analysis and immunohistochemistry. Pregnant Sprague-Dawley rats were evaluated on various time sets: days 10.5 (P10.5), 12.5 (P12.5), 17.5 (P17.5), and 19.5 (P19.5). In Westhern blot analysis, expression of AQP-1, 2 was peaked at P17.5 and AQP-3 at 19.5. Immunoreactivity of AQP-1 of NP rat was detected in the apical membranes of proximal tubules and thin limb of Henle loop. In pregnant rats, the pattern of cellular labeling of AQP-1 protein was identical to NP rat, but signal intensity was continuously increased from P10.5 and peaked at P17.5. In NP rat, immunoreactivity of AQP-2 was the most prominent in apical region and moderate in cytoplasm of the principal cells of entire collecting duct. In pregnant rats, the pattern of cellular labeling of AQP-2 protein was identical to NP rat, but signal intensity was moderately expressed in P10.5 and P12.5 and most prominent signal was observed in P19.5. In NP rat, immunoreactivity of AQP-3 was most prominent in the bosolateral plasma membrane of principal cells of entire collecting duct. In pregnant rats, the pattern of cellular labeling of AQP-3 protein was identical to NP rat, but signal intensity was continuously increased from P10.5 to P17.5 and peaked at P19.5. These results suggest that the expansion of extracellular fluid volume and water retention are regulated by AQP-1, 2, and 3 during the pregnancy, especially at late stage.

Analysis of Hair Protein from Extrinsic Hair Damage / 대한피부과학회지

BACKGROUND: Various kinds of hair products are widely used due to the increase of interest in hair styling. Cosmetic procedures such as permanent waving are very popular today, but the medical studies related to the meaning and restoration pattern of hair damage are mainly based on structural findings. In measuring the degree of hair damage by the moleculobiological methods rather than the structural studies, the findings seem to be highly objective and standardized. OBJECTIVE: The purpose of this study was to identify the patterns of hair damage and restoration through electrophoresis and western blot analysis of hair proteins. METHODS: The three volunteers who we selected as subjects did not have any specific medical illness and had not performed any special cosmetic procedures which could have caused hair damage during the six months before the study. We conducted permanent waving on them. Human hair samples were obtained from the occipital scalp, which were that was not affected by the androgen. We performed extraction and concentration of the whole and partial hair protein, then operated electrophoresis and western blot analysis of the hair protein. RESULTS: In the western blot analysis of whole hair proteins, there was one positive finding on subject A. This may have resulted from the small amount of partial proteins among the whole hair proteins. In the western blot analysis of partial hair proteins, subject A and B showed positive findings. In particular, positive findings were found on the 14th week of the experiment. CONCLUSION: These results show a change in the hair proteins due to hair damage, and ultrastructurally, we found the possibility of prologation of actual hair damage longer than expected.

Expression of Cdc25A, Cdc25B and Cdc25C in cervical carcinoma / 대한산부인과학회지

OBJECTIVE: This study was undertaken to quantitatively detect Cdc25A, Cdc25B and Cdc25C in cervical carcinoma and determine the relationship between the expression of mRNA and protein of cell division cycle (Cdc)25 phosphatase and various clinicopathologic prognostic factors of cervical carcinoma. METHODS: 39 patients diagnosed with cervical carcinoma between February 2000 to March 2005 and 10 patients with benign gynecologic disease were enrolled in this study. A reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were used to analyze the expression of Cdc25 phosphatase mRNA and protein in fresh invasive cervical cancer tissue and normal cervix tissue. RESULTS: The mRNA expressions of Cdc25A, Cdc25B and Cdc25C in the cancer tissues were significantly greater than in the control (p=0.02, 0.01, 0.02), respectively. A Western blot analysis yielded same results (p=0.01, 0.02, 0.01). There were also significant relationships between the age and the Cdc25B mRNA expression (p=0.03), between the cell type and the Cdc25C mRNA expression (p=0.04). However, other clinicopathologic prognostic factors including stage, subtype, SCC Ag level, DNA flow cytometry, lymph node metastasis, lymphovascular space invasion and HPV positivity were not statistically significant. CONCLUSION: Our results show that Cdc25A, Cdc25B and Cdc25C expression levels were significantly greater in cervical cancer patient group than in those of control group. Thus Cdc25 phosphatase might play an important role in carcinogenesis of cervical carcinoma. Further studies based on the correlation between Cdc25 phosphatase and survival rate would be need to support Cdc25 phosphatase as a prognostic factor of cervical carcinoma.

Study on expression of glycosaminoglycan in adenoid cystic carcinoma

Adenoid cystic carcinoma is malignant tumor in salivary gland, and its behavior is very invasive. Of all malignant tumor adenoid cystic carcinoma is occured in frequency of 4.4% in major salivary gland, and 1.29% in minor salivary gland. Histopathologically, adenoid cystic carcinoma is characterized by a cribriform appearance, and tubular form and solid nest type tumor can be seen. The tumor cell structure composed of modified myoepithelial cell, and basaloid cell. Extracellular matrix of this tumor cell contains variable ground substance with basement membrane component. Basement membrane matrix composed of collagen fibers, glycoproteins, proteoglycans, and its function is well known that it participate in differentiation, proliferation, and growth of tumor cell. Basement membrane molecule is essential for invasion of peripheral nerve, blood vessel, skeletal muscle in tumor cell of adenoid cystic carcinoma. In many studies, the tumor cell of adenoid cystic carcinoma containing modified myoepithelial cell participate in synthesis of proteoglycan. In this study, tissue sample of adenoid cystic carcinoma of human salivary gland were obtained from 15 surgical specimen, and all specimen were routinely fixed in 10% formalin and embedded. Serial 4-micrometer thick sections were cut from paraffin blocks. the histopathologic evaluation was done with light microscopy. And, the immunohistochemical staining, characteristics of glycosaminoglycan were observed. For biochemical analysis of glycosaminoglycan, isolation of crude glycosaminoglycan from tumor tissue and Western bolt analysis were carried out. With transmission electomicroscopy, tumor cell were observed. Biologic behavior of adenoid cystic carcinoma was observed with distribution and expression of basement membrane of glycosaminoglycan in tumor cells, The results obtained were as follows: 1. In immunohistochemical study, chondroitin sulfate is postively stained in tumor cell and interstitial space, dermatan sulfate is weakly stained in ductal cell. But keratan sulfate is negatively stained. 2. In immunohistochemical study, heparan sulfate is strong positive stained in tumor cell and basement membrane, especially in invasion area to peripheral nerve tissue. 3. In transmission electromicroscpic view, the tumor cells are composed modifed myoepithelial cells, and contains many microvilli and rough endoplasmic reticulum. 4. In Western blot analysis, the expression of glycosaminoglycan is expressed mostly in heparan sulfate. From the results obtained in this study, tumor cell of adenoid cystic carcinoma is composed modified myoepithelial cell, and glycosaminoglycan of basement membrane molecule of heparan sulfate and chondroitin sulfate mostly participate in the development and invasiveness of adenoid cystic carcinoma by immunohistochemical study and western blot analysis.

Localization of Cyclooxygenase Isozymes in Dermal Wound Healing in Mouse

Cyclooxygenase(COX)-1 and COX-2 expression in dermal wound healing of mouse was detected by immunohistochemistry and Western blot analysis. In order to gain more information on the functional importance of COX-1 and COX-2 in dermal wound healing, we analysed COX-1 and COX-2 protein levels using the Western blotting technique. In addition, we used immunohistochemistry to determine the cellular localization of the protein products. The collected skins were rapidly frozen and kept at -70degrees Cuntil assayed. Each frozen skin was lysed with 0.5 ml of ice-cold solution. Large tissue debris and nuclear fragments were removed by two low-speed centrifugations and the resulting supernatant fraction was used for blots. The skin extracts were stored below -20degrees Cfor further experiments. By Western blotting, compared to the activity of COX-2 in normal skin, its activity was increased at days 1, 4, 8, and 12 and was maximal at 1 day after incisional wound of mouse skin whereas COX-1 was barely detectable. In normal skin, COX-1 immunostaining was observed among the basal cells of epidermis whereas COX-2 immunostaining was detected in the more differentiated, suprabasal keratinocytes. At post-incision 1-4 days, COX-2 staining was particularly prominent in the inflammatory cells, and at day 8, many macrophage-like cells were stained positively. COX-2 immunoreactive fibroblast, macrophage-like cells, and newly formed vascular endothelial cells were increased in number at 12 days after incision. These data suggest that COX-2 is constitutively expressed, just as is COX-1, in epidermis and is associated with keratinocyte differentiation. In addition, these findings support the well-established role for COX-2, the prostaglandins that they generate, as mediators of inflammatory response.

A Comparison Study On Distraction Osteogenesis In The Rat's Mandible According To Distraction Rhythm

Anti-glomerular basement membrane autoantibodies in Chinese patients with rapidly progressive glomerulonephritis and their clinical relevance / 中华肾脏病杂志

Objective To determine the prevalence of autoantibodies against glomerular basement membrane(GBM) in sera of Chinese patients with rapidly progressive glomerulonephritis(RPGN)and to evaluate their clinical relevance. Methods Serum anti-GBM antibodies were detected in 29 RPGN patients by enzyme-linked immunoassay(ELISA)using collagenase solublized human GBM as solid phase ligand. Positive sera were also oonfirmed by Western blot analysis. Results Of the 29 RPGN patients, 5(17%)were positive for anti-GBM autoantibodies. One positive for both anti-GBM autoantibody and ANCA. On Western blot analysis. 23 000～27 000 and 40 000～54 000 polypeptides could be blotted. On direct immunofluoresence there were granular deposits of immunocomplex in capillary loops in three of four. Conclusions The prevalence of anti-GBM antibody mediated RPGN is not rare in China. Using ELISA to detect circulating anti-GBM autoantibodies had been proved to be a more specific and sensitive methods. It is important to detect circulating anti-GBM autoantibodies early for patients with RPGN in order to save time for appropriate therapy.

Expression of Survivin, Anti Apoptosis Gene in Leukemia and Correlation between Expression of Survivin,Fas and Caspase / 实用儿科临床杂志

Objective To explore the relation ship between expression of survivin gene in leukemia cells and its clinical effects, and to study the mechanism of survivin resist-apoptosis function in leukemia.Methods Survivin expression was detected by Western blots analysis and expressions of Fas and Caspase were examined by immunohistochemistry in 18 leukemia patients.Results Thirteen cases in peripheral blood mononuclear cell survivin positive expression was detected in 18 leukemia patients(72.2%), but no survivin expression in 10 normal persons. There were significant difference of survivin expression in ALL/ANLL patients groups and different WBC groups(P

Expression of Survivin in leukemia by Western blot analysis and immunofluorescence method / 西安交通大学学报(医学版)

Objective To compare Western blot analysis and immunofluorescence method in Survivin expression. Methods By using Western blot analysis and immunofluorescence method, Survivin gene expressions of the peripheral blood mononuclear cells in 18 patients with leukemia and 10 healthy persons were analysed. Results Positive expression of Survivin gene was detected in 13 of 18 leukemic patients by Western blot analysis and in 11 of 18 leukemic patients by immunofluorescence method. Negative expression of Survivin gene was detected in 10 healthy persons. The results of Survivin expression were the same in 16 patients by Western blots analysis and immunofluorescence method (consistent rate is 88.89%), but the positive expression of Survivin by Western blot analysis was the negative expression by immunofluorescence method in another 2 patients. Conclusion Survivin gene expressed selectively in leukemic cells, but not in normal mononuclear cell in peripheral blood. It indicated that Survivin expression is effective monitoring index in diagnosis and prognosis of leukemia. Survivin expression by Western blot analysis was consistent with immunofluorescence method.