This study was carried out to investigate the pathological changes with paraquat(1.1'-dimethyl-4.4'-dipyrildiylium dichloride) administered by intraperitoneally, orally, skin applied in mice, rats and rabbits. Results were obtained as follows; In 2 days after paraquat administration clinical signs anorexia, depression, tachypnea, and tachycardia, respiratory failure, coma and death were observed in mice, rats and rabbits. Compared toxicity of paraquat with mouse were observed toward to paraquat that resistance was strong than rats and rabbits. Also, mortality of skin application were found the low than intraperitoneal and high than oral administration. In the case of gross observation were appear lips moisture in orally administered rats and rabbits by skin application. Lung of all laboratory animals were observed congestion and haemorrhage, swelling or atrophy. In the case of microscopic findings were severe congestion and haemorrhage, interstitial pneumonia of lung. Congestion and haemorrhage of liver, congestion and haemorrhage, renal tubule epithelium necrosis of kidney were observed in mice, rats and rabbits. Skin application group of mice, rats and rabbits showed infiltration of inflammatory cells and folliculitis of epidermis and dermis. Also, in oral administration group showed congestion and haemorrhage, tachment, necrosis of alimentary tract mucosa.
During normal early embryonic development in mammals, the global pattern of genomic DNA methylation undergoes marked. changes. The level of methylation is high in male and female gametes. Thus, we cloned the cDNA of the porcine DNA methyltransferase 1 (Dnmt1) gene to promote the efficiency of the generation of porcine clones. In this study, porcine Dnmt1 cDNA was sequenced, and Dnmt1 mRNA expression was detected by reverse transcription-polymerase reaction (RT-PCR) in porcine tissues during embryonic development. The porcine Dnmt1 cDNA sequence showed more homology with that of bovine than human, mouse, and rat. The complete sequence of porcine Dnmt1 cDNA was 4,774-bp long and consisted of an open reading frame encoding a protein of 1611 amino acids. The amino acid sequence of porcine DNMT1 showed significant homology with those of bovine (91%), human (88%), rat (76%), and mouse (75%) Dnmt1. The expression of porcine Dnmt1 mRNA was detected during porcine embryogenesis. The mRNA was detected at stages of porcine preimplantation development (1-cell, 2-cell, 4-cell, 8-cell, morula, and blastocyst stages). It was also abundantly expressed in tissues (lung, ovary, kidney and somatic cells). Further investigations are necessary to understand the complex links between methyltransferase 1 and the transcriptional activity in cloned porcine tissues.
Journal of Physiology & Pathology in Korean Medicine
/
v.23
no.2
/
pp.351-359
/
2009
Etiological studies of diabetes and its complications showed that oxidative stress might play a major role, Therefore, many efforts have been tried to regulate free oxygen radicals for treating diabetes and its complications. Mori Fructus extract has been known to be effective for the antidiabetic, antihyperlipidemic and antiobesitic prescription, and composed of four crude herbs, The aim of this study was to investigate the effect of Mori Fructus extract in male ob/ob mouse with severe obesity, hyperinsulinemia, hyperglycemia, hyperlipidemia. Mice were grouped and treated for 5 weeks as follows. Both the lean (C57BL/6J black mice) and diabetic (ob/ob mice) control groups received standard chow. The experimental groups were fed with a diet of chow supplemented with 7.5, 15 and 30 mg Mori Fructus extract per 1 kg of body weight for 14 days. The effects of Mori Fructus extract on the ob/ob mice were observed by measuring the serum levels of glucose, insulin, lipid components, and the kidney levels of reactive oxygen species (ROS), MDA+HAE, GSH and also the enzyme activities involved in polyol pathway. Western blotting was performed using anti-AGE, anti-RAGE respectively. Mori Fructus extract lowered the levels of serum glucose and insulin in a dose dependent manner. Total cholesterol, triglyceride and free fatty acid levels were decreased, while the HDL-cholesterol level was increased, in Mori Fructus extract treated groups. Renal aldose reductase and sorbitol dehydrogenase activities were increased in the ob/ob mice, whereas those were inhibited in the Mori Fructus extract-administered groups. Mori Fructus extract inhibited the generation of ROS in the kidney. MDA+HAE level was increased and the GSH level was decreased in the ob/ob mice, whereas those were improved in the Mori Fructus extract-administered groups. Mori Fructus extract inhibited the expression of AGE, RAGE in the ob/ob mice. The results suggested that Mori Fructus exerted the antidiabetic and antihyperlipidemic activities by regulating the activities of polyol pathway enzymes, scavenging the ROS, decreasing the MDA+HAE level, increasing the GSH level and inhibiting the expression of AGE, RAGE in the ob/ob mice.
Artemisinin, a new antimalarial to treat patients infected with strains of Plasmodium jalciparum, derived from the plant Artemisia annua Linn, has immunopharmacologic actions such as enhence the PHA -induced lymphocyte transformation rate, increased the weight of spleen but reduced the weight of thymus, reduced phagocytic function of peritoneal macrophage, remarkably reduced the level of serum IgG and hemolysin fonning capacity (sentitized with SRBC), inhibited the activity of Ts cells of donor mice by supraoptimal immunuization(SOI), but enhenced activity of Ts cells of recipient mice by SOI. These results suggested that Ts cells may be the target cells of artemisinin. To the serum complement C3 level of plasmodium berghei-infeted mice, artemisinin (i. m,) could remarkly increase it. The artemisinin also obviously reduced the prostaglandin E(PGE) in the mouse hind paw swelling induced by carrageenin. Numerous studies have demonstrated that pharmacologic doses of PGE attenuate the development of immunocomplex nephritis. Some autologous immune mechanisms may be invoolved In the pathogensis of some types of glomurulonephritis. Glomerular abnormalities can be induced in animals by variety of immunological manipulations. The resulting disorder has many clinical and pathogical similarities to the disease in human. Our purpose was therefore to test the ability of the artemisinin to lessen the severity of rabbit IgG accelerated nephrotoxic serum glomerulonephritis in mice model. Mice which had treated with rabbit IgG and NTS, administrated with saline, showed Significant inceases of urinary protein, cholesterol level, and decrease of serum albumin in NS group. On the contrary, By i.g. adminstration of artemisinin at dose of 12.5, 25 and 50 mg/kg for 14 days after NTS injection, shown that artemisinin inhibited the nephritic changes in some parameters by means of urinary protein(p<0.05, p<0.01) and serum choleterol(p<0.05, p<0.01) and albumin (p<0.05, p<0.01), blood urea nitrogen (p<0.05, p<0.01), serum albumin(p<0.05, p<0.01); Cyclophosphamide(i.p. 10mg/kg for 14d) had almost same effect as the artemisinin had. Morphological studies shown that The picture of kidney from the mouse with NTS-nephritis accerated with rabbit IgG, treated with i.g. saline as the control, the mesangiocapillary were enlarged and proliferated; There were inflammatory cells infiltrating around the glomeruli; The ethelial cell were proliferated in the wall of Bowman's capsule. Histopatholological picture of kidney from the NTS-nephritis accerated with rabbit IgG mouse treated with i.p. 10mg/kg cyclophosphamide as the positive control. No siginicant histopathological evidence were found. Treaded with i.p. 12.5mg/kg artemisinine, the picture shown that mesangiocapillary were lightly proliferated; There were inflammatory cells infiltrating around the glomeruli; Treaded with i.p. 25mg/kg artemisinine, The picture shown that the mesangiocapillary were lightly proliferated; Treaded with i.p. 50mg/kg artemisinine, The picture shown that both the mesangiocapillary proliferated and the inflammatory cells infiltrating around the glomeruli are less than treated with saline, 12.5 and 25 mg/kg artemisinine. On the basis of these studies we conclude that the artemisinin can relieve pathological change caused by NTS-nephritis aacerated with rabbit IgG.
Kim, Kyoung-Shin;Moon, Sung-Sikm;Kim, Byoung-Soo;Kang, Jung-Soo
Journal of Haehwa Medicine
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v.20
no.1
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pp.11-23
/
2011
Systemic Lupus Erythematosus(SLE) is an autoimmune disease invading the skin, joint, kidney, intestinal membrane, neurosystem and other organs. SLE is an autoimmune disease characterized by immune dysregulation resulting in the production of antinuclear antibodies(ANA), generation of circulating immune complexes, and activation of the complement system. In Korean medicine, lupus can be classified as acute arthritis, reddish butterfly erythema, asthenic disease, edema and so on. The cause and procedure of the diseases are flourishing noxious heat, excessive fire due to deficiency of yin, blood stasis due to stagnation of qi, internal movement of the liver-wind, congenital deficiency, exhausted vital-qi, which are treated by clearing away heat and cooling the blood, nourshing yin and extinguishing fire, treating flatulence and activating blood circulation, nourishing the blood to expel wind, invigorating the liver and kidney, invigorating qi and replenishing the blood. To experimentally examine the influence of Insam-Buja-Tang (Ginseng & Aconiti Extract, IBT) on the outbreak and development of lupus, lupus induce MRL/MpJ-Faslpr lupus-prone mice model was used. As IBT was orally administrated to a lupus model mouse, various tests such as the weight, urine protein, renal function, Lymph cell test of the spleen, Cytokine expression, histopathological analysis of kideny were performed to see the influence on the kidney and whether it work effectively on the immune function. The main purpose of this study is to evaluate the effect of IBT on MRL/MpJ-Faslpr lupus-prone mice model. The effect of IBT on MRL/MpJ-Faslpr lupus-prone mice that can have autoimmune disease similar to SLE in human was evaluated after IBT per oral in the present study.
In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney mononuclear cells and partially characterized 7 Mabs in the morphological and mitogen-based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for lCK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK II-I. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright's stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Con A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens.
Ochratoxin A was produced from Aspergillus ochraceus ATCC 18472 which was then orally administered into the experimental mice to study the toxic levels of ochratoxin A. AELISA (enzyme-linked immunosorbent assay) method which is more rapid and safe than conventional analytical method, was developed by using ochratoxin A antibody. This method was successfully used to measure the levels of ochratoxin Ain blood, liver and kidney of mice. In order to produce a large amount of ochratoxin A to study toxicity in the mice, Aspergillus ochraceus ATCC 18412 was incubated in the rice medium and as a result 0.5 g of ochratoxin A form rice medium (kg) was produced after extraction and purification Feed consumption and gain in body weight of mice with ochratoxin A at a level of $10 \mu\textrm{g}/g$ body weight was significantly (p<0.05) reduced as compared with control during period of 3 weeks. Ochratoxin A-BSA conjugate was made by putting 13 mole of ochratoxin A on I mole of BSA. This conjugate was used to develop ELISA method. The minmum detection level of ochratoxin A by established ELISA method was 0.5 ppb. After oral adminstraton of ochratoxin A dose of $10\;\mu\textrm{g}/g$ every two day for 3 weeks, concentration of ochratoxin A was measured in the blood, liver and kidney by ELISA method. The level of ochratoxin A was $11\;\mu\textrm{g}/dl,\;0.9 \mu\textrm{g}/g\;and\;3.7\;\mu\textrm{g}/g$ in the blood, liver and kidney, respectively.
Background: Minor histocompatibility HY antigen, as a transplantation antigen, has been known to cause graft rejection in MHC (major histocompatibility complex) matched donor-recipient. The aim of our study is to investigate the role of male antigen (HY) disparity on MHC matched pancreatic islet transplantation and to examine the mechanism of the immune reaction. Methods: Pancreatic islets were isolated and purified by collagen digestion followed by Ficoll gradient. The isolated islets of male C57BL6/J were transplanted underneath the kidney capsule of syngeneic female mice rendered diabetic with streptozotocine. Blood glucose was monitored for the rejection of engrafted islets. After certain period of time, tail to flank skin transplantation was performed either on mouse transplanted with HY mismatched islets or on sham treated mouse. The rejection was monitored by scoring gross pathology of the engrafted skin. Results: HY mismatched islets survived more than 300 days in 14 out of 15 mice. The acceptance of second party graft (male B6 islets) and the rejection of third party graft (male BALB/c islets) in these mice suggested the tolerance to islets with HY disparity. B6 Skin with HY disparity was rejected on day $25{\pm}7$. However, HY mismatched skin transplanted on the mice tolerated to HY mismatched islets survived more than 240 days. Tetramer staining in these mice indicated the CTL recognizing MHC Db/Uty was not deleted or anergized. Conclusion: The islet transplantation across HY disparity induced tolerance to HY antigen in C57BL6 mouse, which in turn induced tolerance to HY mismatched skin, which otherwise would be rejected within 25 days. The MHC tetramer staining suggested the underlying mechanisms would not be clonal deletion or anergy.
We have cloned a novel mouse zinc finger protein gene Znf313 by rapid amplification of cDNA ends (RACE) according to the homologue of human ZNF313 gene. The cDNA is 2,163 base pairs (bp) in length and encodes a 229 amino acids (aa) protein with a $C_3HC_4$ ring finger domain and three $C_2H_2$ domains. 89% and 93% nucleotide (nt) and aa sequence identity is observed with its human homologue. Revealed by Northern blot and RT-PCR, full mRNA consists of 2.16 kb and widely expresses in tissues as a single transcript, most abundantly in heart, liver, kidney and testis. The expression of Znf313 in testis is detected in all development stages. Western blot analysis also reveals that Znf313 is expressed in the tissues. Immunohistochemical staining and subcellular localization demonstrate that Znf313 is expressed both in the cytoplasm and nucleus whereas predominantly localized in the nucleus. Present data suggests that Znf313 gene might play a fundamental role in gene transcription and regulation in organism and relates to spermatogenesis.
Flounder (Paralichthys olivaceus) Immunoglobulins (Igs) were purified from the serum of mouse IgG-immunized flounder by using affinity chromatography. Under denaturing conditions in SDS-PAGE, the flounder Igs appeared to be composed of 2 heavy (H) chains (72 and 77 kDa) and two light (L) chains (26 and 28 kDa). Monoclonal antibodies (MAbs) were produced by the fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells that were previously sensitized against affinity-purified flounder Igs. In a Western blot analysis, the produced MAbs, FIM511, FIM519, and FIM562 recognized both the 72 and 77 kDa H chains, 26 kDa, and 28 kDa L chain, respectively. Mouse antiserum against flounder Igs reacted more strongly with the L chain of 28 kDa than with 26 kDa, suggesting that the 28 kDa molecule is more immunogenic than the 26 kDa L chain molecule. In a FACS analysis, the ratios of the Ig+ cell population in the flounder head kidney and spleen cells were 49% and 24%, respectively. Unexpectedly, however, the ratios of the Ig+ B-like cell population in the flounder were not significantly augmented, even after the immunization of an immunogenic antigen. This suggests that the humoral immune response in fish could be considerably different from that in mammals. The produced MAbs in this study would be useful in characterizing flounder Ig+ B-like cells and in developing flounder Ig detecting an immunoassay system.
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