• Title/Summary/Keyword: granulocytes

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Production of the Novel Disease Animal Model by Used Tet-off System

  • Park, Jun-Hong;Kim, Kil-Soo;Lee, Eun-Ju;Kim, Myoung-Ok;Kim, Sung-Hyun;Kyoungin-Cho;Jung, Boo-Kyung;Kim, Hee-Chul;Sol ha Hwang
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.54-54
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    • 2003
  • The activation of protooncogenes or the inactivation of their gene products may be a specific and effective functional study for human neoplasia. To examine this possibility, we have used the tetracycline regulatory system to generate transgenic mice that conditionally express the HccR-2 protooncogene in vivo. The new human cervical cancer protooncogene (HccR-2) was detected from cervical cancer cell line. To elucidate its biological functions, we generated transgenic mice that expressed the HccR-2 gene. The sustained expression of the HccR-2 transgene culminated chronic neutrophilic leukemia (CNL). CNL is a rare chronic myeloproliferative disorder that presents as a sustained, mature neutrophilic leukocytosis with few or no circulating immature granulocytes, the absence of peripheral blood monocytosis, basophilia, or eosinophilia, and infiltration of neutrophils at the liver, spleen and kidney. Mice expressing the HccR-2 and tetracycline-transactivating protein (tTa) transgene were found to have altered myeloid development that was characterized by increased percentages of mature neutrophil and band form neutrophil in the peripheral blood, liver and spleen. Activation of the transgene causes CNL. In our model, expression of HccR-2 transgene mice was similar in many respects to the human CNL. This model will be valuable not only for investigating the biological properties of the HccR-2 and other protooncogenes in vivo but also for analyzing the mechanism involved in the progression of CNL.

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Genome-wide DNA methylation pattern in a mouse model reveals two novel genes associated with Staphylococcus aureus mastitis

  • Wang, Di;Wei, Yiyuan;Shi, Liangyu;Khan, Muhammad Zahoor;Fan, Lijun;Wang, Yachun;Yu, Ying
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.203-211
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    • 2020
  • Objective: Staphylococcus aureus (S. aureus) is one of the major microorganisms responsible for subclinical mastitis in dairy cattle. The present study was designed with the aim to explore the DNA methylation patterns using the Fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) techniques in a S. aureus-infected mouse model. Methods: A total of 12 out-bred Institute of Cancer Research female mice ranging from 12 to 13 weeks-old were selected to construct a mastitis model. F-MSAP analysis was carried out to detect fluctuations of DNA methylation between control group and S. aureus mastitis group. Results: Visible changes were observed in white cell counts in milk, percentage of granulocytes, percentage of lymphocytes, CD4+/CD8+ ratio (CD4+/CD8+), and histopathology of mice pre- and post-challenge with S. aureus. These findings showed the suitability of the S. aureus-infected mouse model. A total of 369 fragments was amplified from udder tissue samples from the two groups (S. aureus-infected mastitis group and control group) using eight pairs of selective primers. Results indicated that the methylation level of mastitis mouse group was higher than that in the control group. In addition, NCK-associated protein 5 (Nckap5) and transposon MTD were identified to be differentially methylated through secondary polymerase chain reaction and sequencing in the mastitis group. These observations might play an important role in the development of S. aureus mastitis. Conclusion: Collectively, our study suggests that the methylation modification in Nckap5 and transposon MTD might be considered as epigenetic markers in resistance to S. aureus-infected mastitis and provided a new insight into S. aureus mastitis research in dairy industry and public health.

Histopathological Studies of Mice after Administration of Radioactive Iodine($^{131}I$) (방사성동위원소옥소(放射性同位元素沃素)($^{131}I$)가 "마우스"의 주요장기(主要臟器)의 병리조직상(病理組織像)에 미치는 영향(影響)에 관(關)한 연구(硏究))

  • Ro, Chae-Song;Lee, Mun-Ho
    • The Korean Journal of Nuclear Medicine
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    • v.1 no.2
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    • pp.85-97
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    • 1967
  • Histopathological changes of various organs of the mice after intra-peritoneal injections of radioactive iodine ($^{131}I$) were experimentally observed. Sixity healthy female mice, weighing average 25 gm, devided into 6 groups, were used. The various doses of $^{131}I$ were injected intraperitoneally at different intervals. The histopathological changes after these treatments were observed in organs such as thyroids, parathyroids, livers, kidneys and gonads. Following were the results; 1) Thyroid: In the group A given $^{131}I$ with a single dose of $10{\mu}C$ per gm body weight, it was observed that the protoplasms of follicular epithelial cells were destroyed, the nuclei were expanded or dissoluted, showing pyknotic changes of nuclei and vacuolizations of protoplasms. In the group B given $^{131}I$ with a single dose of $5{\mu}C$ per gm body weight, hyperemias, hemorrhages and hyaline degenerations in the whole area were observed. In the group C given $^{131}I$ with 3 doses of $2.5{\mu}C$ per gm body weight every week, the thyroid parenchyms were destroyed and epithelial cells of varing size were observed in the fibrinous tissues. In the group D given $^{131}I$ with 6 doses of $0.5{\mu}C$ per gm body weight every week, some destroyed follicles and new borne follicles were observed. But the histopathological changes resemble the follicles of the normal thyroid gland. In the group E and F given $^{131}I$ with 8 and 10 doses of $0.2{\mu}C\;and\;0.01{\mu}C$ for each group per gm body weight every two days, both pyknotic changes of nuclei and cytoplasmic vacuolizations of the follicular epithelia, hypertrophies of follicles and abnormal irregular follicular structures were observed, and in the group F, lymphocytes appeared around the thyroid glands. 2) Parathyroid: In the group A, hyperemia, proliferations of connective tissues, karyorrhexes and vacuolizations were observed. In other experimental groups, no particular pathological change was observed. 3) Liver: The degnerative changes and acute or chronic inflammatory changes were observed in proportion to the amount of $^{131}I$ injected. Atrophies of the liver cells, dilatations of sinusoids, hyaline degenerations and necrotic pictures were observed. 4) Kidney: In the group A, congestions and infiltrations of mononuclear cells and granulocytes were observed around the cortical arteries, and in the group B, the degenerative changes of cortexes, and, in the group C and D, hydronephrotic changes were observed respectively, and hyaline degenerations were partially observed. 5) Gonad: In the group A, the follicles were degenerated. The ova in the follicles showed irregular figures. The changes in the group B were almost the same as in the group A, but the changes were mild. In the group C, the destructions of whole ova, the hypertrophies of ovarian follicular membranes and pyknotic changes of nuclei were observed. In the group D, the pathological changes were similar to that of group C, but mild in the grade. In the group E, almost none of ovarian follicular fluid was observed, and in the group F, the tissue pictures were almost similar to that of the normal group.

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Virulence of Ornithobacterium rhinotracheale Isolates for Embryonated SPF Eggs and Broilers (국내에서 분리한 Ornithobacterium rhinotracheale 균의 종란과 육계에서의 병원성)

  • Kwon, Yong-Kuk;Jeon, Woo-Jin;Kang, Min-Soo;Oh, Jae-Young;An, Byung-Ki;Song, Eun-A;Kwon, Jun-Hun;Lee, Cheong-San;Kim, Jae-Hong
    • Korean Journal of Poultry Science
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    • v.37 no.2
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    • pp.159-165
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    • 2010
  • Field strains of Ornithobacterium rhinotracheale (OR) were tested on their virulence in specific pathogen free (SPF) embryonated chicken eggs and 3-week-old broilers. When infected with three different OR isolates (OR-161, OR-240 and OR-295) through yolk sac infection route, all strains appeared to be highly pathogenic with responsible mortality 66% and 100% within 12 days post infection (DPI). To test the virulence of OR in the commercial broilers, 3 week-old broilers were grouped depends on the inoculation route of OR isolate (OR-295) through five different infection routes; group 1 (IT: intratracheal), group 2 (IM: intramuscular), group 3 (IV: intravenous), group 4 (aerosol) and group 5 [Mixed: NDV (LaSota)+OR aerosol]. Within 5 to 7 days after inoculation, only broilers given NDV+OR were slightly depressed and coughing, and had mild facial redness. Grossly, foamy and yellow-white yogurt like exudate in the air sacs, predominantly in the abdominal air sacs was present. In histology, infiltration of the air sac epithelium and lamina propria by macrophage and polymorphonuclear granulocytes was seen with cell debris and inflammatory cells, correlated with the presence of OR antigen, as demonstrated by immunohistochemistry. Field strains of OR were able to induce high mortality in the embryonated chicken eggs, whereas broilers were less susceptible to OR infection. Interestingly, in the absence of NDV infection, the four groups of OR single infection only different route showed minimal and temporary microscopic air sac lesions. Thus, Newcastle disease virus (LaSota strain) showed triggering effects on the OR infection in chickens.

Damage of radioprotection and antitumor effects of water-soluble propolis

  • Terai, Kaoru;Ryu, Myung-Sun;Itokawa, Yuka;Maenaka, Toshihiro;Nakamura, Takashi;Hasegawa, Takeo;Choi, In-Suk;Ishida, Torao;Gu, Yeun-Hwa
    • Advances in Traditional Medicine
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    • v.6 no.1
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    • pp.12-20
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    • 2006
  • Some natural products are able to inhibit radiation effects and exert an antitumor effect with fewer adverse reactions; however, their antitumor effects are less than those of widely-used synthetic drugs. Propolis is a natural material that has been attracting attention, and we extracted this material with water and investigated the effect of continuous propolis administration on radioactivity-induced reduction of hemocytes, in addition to the antioxidant and antitumor effects of propolis. Following a 1-week adjustment period, water-soluble propolis was administered intraperitoneally to male ICR mice at a dose of 100 mg/kg every other day for 2 weeks. Following administration, 2 Gy whole-body irradiation was performed and the counts of leukocytes, lymphocytes, and granulocytes and monocytes in the peripheral blood were determined 1, 3, 7, 15 and 30 days after irradiation. These cells were considered since they are closely associated with immunity to radioactivity. In a second experiment, water-soluble propolis was similarly administered to the mice for 2 weeks after a 1-week adjustment period, and 2 Gy whole-body irradiation was performed. The antioxidant effects in hemocytes were then investigated using 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), a radical generator. In a third experiment, $1\;{\times}\;10^6$ Sarcoma-180 cells were inoculated into the right thigh of mice, which were divided into four groups: control, water-soluble propolis-treated, 6 Gy irradiated and water-soluble propolis-treated + 6 Gy irradiated groups, and changes in tumor size were measured for 20 days. Statistical analysis was conducted using ANOVA for multiple groups. In the three experiments, administration of water-soluble propolis inhibited the reduction of hemocytes caused by whole-body irradiation, showed antioxidant effects against radioactivity, and inhibited tumor growth, respectively. In conclusion, our data suggest that the antioxidant effect of watersoluble propolis inhibits hemocyte reduction caused by whole-body irradiation and enhances immunological inhibition of tumor growth.

Effects of SPDJTK(SoPungDoJeokTangKami) and Concurrent Administration of AJ (Atopy cream, Jawoongo) Plus SPDJTK on Atopic Dermatitis-like Skin Lesions in NC/Nga Mouse Induced by BMAC (아토피양(樣) 피부염 NC/Nga 생쥐에서 소풍도적탕가미(消風導赤湯加味)와 아토피크림, 자운고(紫雲膏) 및 소풍도적탕가미(消風導赤湯加味)의 병용투여가 피부염에 미치는 영향)

  • Han, Dal-Soo;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.24 no.1
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    • pp.9-35
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    • 2010
  • Objectives The purpose of this study is to investigate the effect of SPDJTK(SoPungDoJeokTangKami) and concurrent administration of AJ(Atopy cream, Jawoongo)+SPDJTK on atopic dermatitis-like skin lesions by using in NC/Nga atopic dermatitis mouse induced by BMAC-induced mice. Methods Clinical skin score, hematology and Serum total IgE and IgG1 of NC/Nga atopic dermatitis mice were evaluated. Moreover, the cytokine level, total cell number, Immunohistochemical staining and Histological features of axillary lymph node(ALN), draining lymph node(DLN), peripheral blood mononuclear cells(PBMCs) and dorsal skin tissue were used in NC/Nga mice. Results Orally administrated SPDJTK with concurrent administration of SPDJTK and AJ decreased the clinical skin score, total cell number of WBC, eosinophils in blood, serum total IgE & IgG1, IL-5, IL-13, IFN-$\gamma$. Also, total cell number of ALN and dorsal skin tissue, absolute cell number of CD4+, CD8+, CD3+CD69+, CD3+CCR3+, CCR3+, CD4+CXCR5+ in ALN, absolute cell number of CD3+CCR3+, CCR3+ in DLN, granulocytes in PBMCs, activation cell number of CD3+CD69+, CCR3+, total cell number of CD3+ T cell in dorsal skin tissue were significantly decreased. Furthermore, thickness of epidermis, infiltrated inflammatory immune cell and mast cell in dermis, amount of Eotaxin2 mRNA, CCR3 mRNA in dorsal skin tissue, gene expression of IL-5, IL-13 mRNA in ALN, CD4+ Th cell in dorsal skin tissue and CCR3+ eosinophils in ALN were all significantly decreased. However, total number of DLN, absolute number of CD3e+ T cell and CD19+ B cell, absolute number of CD4+, number of Th cell in DLN and gene expression of foxp3 mRNA were significantly increased significantly. Conclusions Concurrent administration of SPDJTK and AJ on atopic dermatitis in NC/Nga atopic dermatitis mouse was very effective treatment for atopic dermatitis.

Inflammation Scan Using $^{99m}Tc-HMPAO$ Labelled Leukocytes ($^{99m}Tc-HMPAO$를 이용한 자가백혈구표지 및 그를 이용한 염증병소의 스캔)

  • Yang, Woo-Jin;Chung, Soo-Kyo;Shinn, Kyung-Sub;Bahk, Yong-Whee;Kim, Hoon-Kyo
    • The Korean Journal of Nuclear Medicine
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    • v.23 no.2
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    • pp.219-223
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    • 1989
  • Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.

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Anti-leukemic Effects of Hwoangbaec-tang in Human Promyelocytic Leukaemia, HL-60 Cells (항백탕 추출물의 인간 백혈병 세포주 HL-60에서 항백혈병 효과)

  • Kim, Young-Jun;Jeon, Byung-Hun;Ju, Sung-Min;Lee, Jang-Chun;Park, Yang-Koo;Lee, Sang-Hyun;Chon, Young-Gyun;Hwang, Joo-Min;Lim, Dae-Hwan;Yun, Young-Gab
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.3
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    • pp.633-639
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    • 2005
  • The composition of Hwoangbaec-tang has been traditionally used in Korea to treat cancer. Hwoangbaec-tang I is the water extracts prepared from Angelica dahurica, Fritillariae verticillata, Ailanthus altissima, Viscum coloratun, Scutellaria Radix, Ginseng Radix, Astragalus membranaceus, and Glycyrrhizae Radix. Hwoangbaec-tang II also is the water extracts prepared from Ginseng Radix, Astragalus membranaceus, and Glycyrrhizae Radix. The anti-leukemic effects of human promyelocytic leukaemia (HL-60 cells) by Hwoangbaec-tang I or II was accessed by propidium iodide staining flow cytometric analysis, and apoptosis-inducing activity was further confirmed by a nuclear morphological change, a ladder pattern of DNA fragmentation, and an activation of caspase-3 and 9. Hwoangbaec-tang I was found to induce the apoptosis of HL-60 cells via caspase-3 and 9 pathway. In the other side, Hwoangbaec-tang II was found to inhibit the growth of HL-60 cells by inducing these cells to differentiate toward granulocytes. These results indicate that the different anti-leukemic effects of Hwoangbaec-tang in HL-60 cells can be induce the apoptosis or differnetiation of HL-60 cells in Hwoangbaec-tang composition dependent manner.

Treatment with Rutin - A Therapeutic Strategy for Neutrophil-Mediated Inflammatory and Autoimmune Diseases - Anti-inflammatory Effects of Rutin on Neutrophils -

  • Nikfarjam, Bahareh Abd;Adineh, Mohtaram;Hajiali, Farid;Nassiri-Asl, Marjan
    • Journal of Pharmacopuncture
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    • v.20 no.1
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    • pp.52-56
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    • 2017
  • Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.

Expression and Production of Human Granulocyte Colony Stimulating Factor (G-CSF) in Silkworm Cell Line (누에세포를 이용한 인간 G-CSF의 발현 및 생산)

  • Park, Jeong-Hae;Jang, Ho-Jung;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1577-1581
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    • 2010
  • Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.