Un Chol Shin;Seoku Bae;Suk-man Kim;Min-Woo Lee;Han Sang Jin;Hyun Park;Kyo Chul Lee;Jung Young Kim;Ji Woong Lee
Journal of Radiopharmaceuticals and Molecular Probes
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v.7
no.2
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pp.105-111
/
2021
89Zr is a positron-emitting radioisotope, which has known as well-suited radioisotope for use in a monoclonal antibody-based imaging agent for immuno-PET. The purpose of this study was to quantitatively evaluate the diagnostic ability of general trastuzumab and thio-trastuzumab as HER2 positive receptors based on Df hexadentate iron chelator. Desferrioxamine-p-SCN (Df-Bz-NCS) and desferroixamine-maleimide (Df-Mal) were purchased from Macrocyclics (Dallas, TX, USA). The trastuzumab was purchased from Roche (Schweiz), and thio-trastuzumab was obtained from professor Hyo-Jeong Hong group (Kangwon National University). The radioisotope 89Zr was produced by domestic purification system and KIRAMS using medical cyclotron (50 MeV, Scantronix). The conjugates of Df-trastuzumab and Df-thio-trastuzumab were prepared with Df-Bz-NCS and Df-Mal under basic aqueous solution (pH 8-9) at room temperature, respectively. The conjugates purified by PD-10 column were mixed with dried 89Zr chloride. 89Zr-labeled conjugates were purified and concentrated by Amicon ultra centrifugal filter. The preparation step and time of 89Zr-labeled conjugates was shorted as 4 steps within 2 hours. 89Zr-labeled conjugates showed the highly radiochemical purity of over 98%, and were very stable until 7 days by the analysis of radio-ITLC method. Each radio-labeled conjugates were also exhibited the highly stability in both PBS buffer and mouse serum. Immuno-PET imaging of 89Zr-labeled conjugates in mice bearing gastric cancer xenograft tumors with HER2 expression showed high tumor uptake in the NCI-N87 HER2-expressing. However, 89Zr-Df-Mal-thio-trastuzumab showed a relatively lower tumor-to-background ratio than 89Zr-Df-Bz-trastuzumab, as well as whole-body distribution. In the results, 89Zr-Df-Bz-trastuzumab was evaluated to have a relatively higher HER2 diagnostic ability than 89Zr-Df-Mal-thio-trastuzumab.
Kim, Ji-Young;Yoon, Seok-Joo;Park, Han-Jin;Kim, Yong-Bum;Cho, Jae-Woo;Koh, Woo-Suk;Lee, Michael
Toxicological Research
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v.23
no.1
/
pp.55-63
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2007
Diethylnitrosamine (DEN) is a nitrosamine compound that can induce a variety of liver lesions including hepatic carcinoma, forming DNA-carcinogen adducts. In the present study, microarray analyses were performed with Affymetrix Murine Genome 430A Array in order to identify the gene-expression profiles for DEN and to provide valuable information for the evaluation of potential hepatotoxicity. C57BL/6NCrj mice were orally administered once with DEN at doses of 0, 3, 7 and 20 mg/kg. Liver from each animal was removed 2, 4, 8 and 24 hrs after the administration. The histopathological analysis and serum biochemical analysis showed no significant difference in DEN-treated groups compared to control group. Conversely, the principal component analysis (PCA) profiles demonstrated that a specific normal gene expression profile in control groups differed clearly from the expression profiles of DEN-treated groups. Within groups, a little variance was found between individuals. Student's t-test on the results obtained from triplicate hybridizations was performed to identify those genes with statistically significant changes in the expression. Statistical analysis revealed that 11 genes were significantly downregulated and 28 genes were upregulated in all three animals after 2 h treatment at 20 mg/kg. The upregulated group included genes encoding Gdf15, JunD1, and Mdm2, while the genes including Sox6, Shmt2, and SIc6a6 were largely down regulated. Hierarchical clustering of gene expression also allowed the identification of functionally related clusters that encode proteins related to metabolism, and MAPK signaling pathway. Taken together, this study suggests that match with a toxicant signature can assign a putative mechanism of action to the test compound if is established a database containing response patterns to various toxic compounds.
The immune-stimulating activities of Bordetella bronchiseptica antigens containing dermonecrotoxin (BBD) loaded in chitosan microspheres (CMs) have already been reported in vitro and in vivo with a mouse alveolar macrophage cell line (RAW264.7) and mice. Therefore, this study attempted to demonstrate the successful induction of mucosal immune responses after the intranasal administration of BBD loaded in CMs (BBD-CMs) in colostrum-deprived pigs. The BBD was introduced to the CMs using an ionic gelation process involving tripolyphosphate (TPP). Colostrum-deprived pigs were then directly immunized through intranasal administration of the BBD-CMs. A challenge with a field isolate of B. bronchiseptica was performed ten days following the final immunization. The BBD-specific IgG and IgA titers, evident in the nasal wash and serum from the vaccinated pigs, increased with time (p<0.05). Following the challenge, the clinical signs of infection were about 6-fold lower in the vaccinated pigs compared with the nonvaccinated pigs. The grades for gross morphological changes in the turbinate bones from the vaccinated pigs were also significantly lower than the grades recorded for the nonvaccinated pigs (p<0.001). Therefore, the mucosal and systemic immune responses induced in the current study would seem to indicate that the intranasal administration of BBD-CMs may be an effective vaccine against atrophic rhinitis in pigs.
In our previous study, we observed that hydrosalpingeal fluid (HSF) adversely effect mouswe embryo development and hatching. The aim of this study was to evaluate the effect of HSF as assessed by the blastocyst development rate (BDR) and by cell counting in vitro. HSF was collected from ninie patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and $T6{\pm}0.4%$ bovine serum albumin were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in micro drops prepared from each of these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The mean BDR in two control media were 88.89% and 85.40%. The mean BDR in media containing 10%, 50%, 100% HSF were 85.87%, 89.58% and $75.57%^*$, respectively ($^*$: p<0.05). The overall mean cell count $({\pm}SEM)$ in control media were $87.6{\pm}9.65\;and\;90.12{\pm}11.38$. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. Mean cell counts were decreased significantly only in blastocysts cultured 100% HSF ($63.8{\pm}13.66$; p<0.01) but not in blastocysts cultured in 10% or 50% HSF ($91.3{\pm}12.44\;and\;82.9{\pm}18.27$, respectively). Thus, it is concluded that HSF has no embyotoxic effect but has a mildly negatively effect on embryonic growth and development.
Lee, Jin A;Lee, Tae Jong;Kim, Jin Young;Shin, Mi-Rae;Park, Hae-Jin;Roh, Seong-Soo
The Korea Journal of Herbology
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v.37
no.5
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pp.63-74
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2022
Objective : Gastritis refers to an inflammatory disease of the gastric mucosa. Alcohol is one of the main aggression factors, causing bleeding and inflammation in the gastric mucosa and it is known to not only increase lipid peroxide levels, but also deplete key antioxidant factors. The purpose of this study was to determine the effect of Uncariae Ramulus et Uncus water extract (URW) in alcohol-induced gastritis. Methods : The total polyphenol and flavonoid contents of URW were confirmed through an in vitro experiment. Also, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity and ferric reducing antioxidant power (FRAP) activity were confirmed. For in vivo experiments, mice were divided into 4 groups (n=8). Also, 1 hr after oral administration of each drug, 50% ethanol was orally administered to induce gastritis. Results : As a result of in vitro experiments, URW showed excellent antioxidant activity. In alcohol-induced gastritis, URW alleviated the damage to the gastric mucosa caused by alcohol. Also, URW decreased reactive oxygen species (ROS) and malondialdehyde (MDA) levels in serum and gastric tissues, and significantly decreased the expression of NADPH oxidases in gastric tissues. In addition, it significantly modulated the nuclear factor erythroid-derived 2-related factor 2 (Nrf2) and nuclear factor-𝜅B p65 (NF-𝜅B) pathways as well as significantly increased the expression of anti-inflammatory proteins. Conclusions : These results suggest that URW not only reduces oxidative stress through excellent antioxidant activity but also relieves gastric mucosal inflammation as a regulator of Nrf2 and NF-𝜅B pathways.
Recent study showed that T cells in the immune organs and peripheral blood are influenced by estradiol, leading to a dysfunction of the immune system. However, little is known about the thymic-gonadal relationship during the estrous cycle in mouse. Therefore, the purpose of this study was to elucidate the mechanism by which a change in estradiol levels during the estrous cycle regulates the development of T cells in the mouse thymus. Six-week-old ICR mice were used and divided into four groups, including diestrous, proestrous, estrous, and metestrous. We first confirmed that ER-${\alpha}$ and - ${\beta}$ estrogen receptors were expressed in thymic epithelial cells, showing that their expression was not different during the estrous cycle. There was also no significant difference in thymic weight and total number of thymocytes during the estrous cycle. To determine the degree of thymocyte differentiation during the estrous cycle, we analyzed thymocytes by flow cytometry. As a result, the percentage of CD4+CD8+ double-positive (DP) T cells was significantly decreased in the proestrous phase compared to the diestrous phase. However, CD4+CD8- or CD4-CD8+ (SP) T cells were significantly increased in the proestrous phase compared to the diestrous phase. In addition, the percentage of CD44+CD25- (DN1) T cells was significantly decreased in the estrous phase compared to other phases, whereas the percentages of CD44+CD25+ (DN2), CD44-CD25+ (DN3), and CD44-CD25- (DN4) were not changed during the estrous cycle. These results indicate that the development of thymocytes may arrest in the DP to SP transition stage in the proestrous phase displaying the highest serum level of estradiol. This study suggests that a change in estradiol levels during the estrous cycle may be involved in the regulation of thymocyte differentiation in the mouse thymus.
Objective: Liver fibrosis is a highly conserved wound-healing response and the final common pathway of chronic inflammatory injury. This study aimed to evaluate the potential anti-fibrotic effect of the combination of Rhei Radix et Rhizoma water extract (RW) and silymarin in a thioacetamide (TAA)-induced liver fibrosis model. Methods: The liver fibrosis mouse model was established through the intraperitoneal injection of TAA (1 week 100 mg/kg, 2-3 weeks 200 mg/kg, 4-8 weeks 400 mg/kg) three times per week for eight weeks. Animal experiments were conducted in five groups; Normal, Control (TAA-induced liver fibrosis mice), Sily (silymarin 50 mg/kg), RSL (RW 50 mg/kg+silymarin 50 mg/kg), and RSH (RW 100 mg/kg+silymarin 50 mg/kg). Biochemical analyses were measured in serum, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), and ammonia levels. Liver inflammatory cytokines and fibrous biomarkers were measured by Western blot analysis, and liver histopathology was evaluated through tissue staining. Results: A significant decrease in the liver function markers AST and ALT and a reduction in ammonia and total bilirubin were observed in the group treated with RSL and RSH. Measurement of reactive oxygen species and MDA revealed a significant decrease in the RSL and RSH administration group compared to the TAA induction group. The expression of extracellular matrix-related proteins, such as transforming growth factor β1, α-smooth muscle actin, and collagen type I alpha 1, was likewise significantly decreased. All drug-administered groups had increased matrix metalloproteinase-9 but a decreasing tissue inhibitor of matrix metalloproteinase-1. RSL and RSH exerted a significant upregulation of NADPH oxidase 2, p22phox, and p47phox, which are oxidative stress-related factors. Furthermore, pro-inflammatory proteins such as cyclooxygenase 2 and interleukin-1β were markedly suppressed through the inhibition of nuclear factor kappa B activation. Conclusions: The administration of RW and silymarin suppressed the NADPH oxidase factor protein level and showed a tendency to reduce inflammation-related enzymes. These results suggest that the combined administration of RW and silymarin improves acute liver injury induced by TAA.
Yu Chun Cai;Chun Li Yang;Peng Song;Muxin Chen;Jia Xu Chen
Parasites, Hosts and Diseases
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v.62
no.1
/
pp.53-63
/
2024
The intracellular parasite Babesia microti is among the most significant species causing human babesiosis and is an emerging threat to human health worldwide. Unravelling the pathogenic molecular mechanisms of babesiosis is crucial in developing new diagnostic and preventive methods. This study assessed how priming with B. microti surface antigen 1 (BHSA 1) and seroreactive antigen 5-1-1 (BHSA 5-1-1) mediate protection against B. microti infection. The results showed that 500 ㎍/ml rBMSA1 and rBMSA5-1-1 partially inhibited the invasion of B. microti in vitro by 42.0±3.0%, and 48.0±2.1%, respectively. Blood smears revealed that peak infection at 7 days post-infection (dpi) was 19.6%, 24.7%, and 46.7% in the rBMSA1, rBmSA5-1-1, compared to the control groups (healthy mice infected with B. microti only), respectively. Routine blood tests showed higher white blood cell, red blood cell counts, and haemoglobin levels in the 2 groups (BMSA1 and BMSA5 5-1-1) than in the infection control group at 0-28 dpi. Moreover, the 2 groups had higher serum interferon-γ, tumor necrosis factor-α and Interleukin-17A levels, and lower IL-10 levels than the infection control group throughout the study. These 2 potential vaccine candidate proteins partially inhibit in vitro and in vivo B. microti infection and enhance host immunological response against B. microti infection.
Park, Hee-Juhn;Nam, Jung-Hwan;Jung, Hyun-Ju;Kim, Won-Bae;Park, Kwang-Kyun;Chung, Won-Yoon;Choi, Jong-Won
Korean Journal of Pharmacognosy
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v.36
no.4
s.143
/
pp.318-323
/
2005
The leaves (KPL) of Kalopanax pictus (KP) are used as a vegetable or a functional food in Korean society. The stem bark (Kalopanacis Cortex, KPS) has been traditionally used to treat neurotic pain, rheumatoid arthritis and diabetic disease. This research was undertaken to demonstrate that the leaf extract of KP (KPL) has also the antinociceptive and antiinflammatory effects like the extract (KPS) of Kalopanacis Cortex and to compare the activity levels of several extracts obtained from KP. Antinociceptive and antiinflammatory effects were measured against the extracts described as followings; KPL-1 (the MeOH extract obtained from the leaf shoot of KP collected on May), KPL-2 (the MeOH extract from KP collected on June), KPL-3 (the MeOH ectract from KP with no thorns), KPS-1 (MeOH extract from KPS of a Korean habitat), KPS-2 (MeOH extract from KPS of a Chinese habitat). The antimociceptive test undertaken by acetic acid-induced writhing, hot plate-, and tail-flick methods using mice. The anti-inflammatory test was also undertaken by measuring the edema in the carrageenan-induced test. The order of activity potency in the antinociceptive and antiinflammatory assays was commonly shown as followings: KPL-3>KPS>1>KPS-2>KPL-1>KPL-2. This order was also observed in acetic acid-induced vascular permeability test. The antiinflammatory activity in carrageenan-induced assay was also observed as the following order: KPL-3>KPS- 1>PS-2>KPL-1>KPL-2. In addition, adjuvant-induced rats were used for a model to assess the oxidative stress. Treatment of the rat with the extracts reduced serum thiobarbituric acid-reactive substances (TBARS), hydroxy radical(OH) and superoxide dismutase(SOD) activity caused by FCA together together with the inhibition of hepatic TBARS level and lipofuscin content. The above finding suggests that the leaf extract has the antinociceptive and antinflammatory activity. It is also suggested that KPL-3 with more potent activity than other tested extracts could be developed for a new available biomaterial.
Rubus coreanus and Astragalus membranaceus Bunge have been used as a traditional medicine with various pharmacological properties for a long time in Asia. We investigated the proliferative and differentiative effects on osteoblastic cells, MG-63 in various mixture ratios of immature fructus extracts of R. coreanus and root extracts of A. membranaceus. With preliminary in vivo ovariectomized mice, we confirmed the effects of the supplementation of various mixture on alkaline phosphatase (ALP) activity in serum. The ratio of 7:3 is considered as the most effective ratio for the proliferation of osteoblastic cells as they increased by 56.8% similar to estradiol. ALP activity in osteoblast increased by the combination of R. coreanus and A. membranaceus and peaked in the ratio of 72:28 and 68:32 respectively. These results indicate that the optimal combination of R. coreanus and A. membranaceus in 7:3 has the most beneficial effects on the activation of osteoblast cells, and give further possibilities that it can be used clinically in the treatment of osteoporosis.
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