• Title/Summary/Keyword: Normal mice

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Skin elasticity improvement effect of Young persimmon and Heated young persimmon by decreased Advanced glycation end products(AGEs) (떫은감과 포제 떫은감의 최종당화산물 생성 억제를 통한 피부 탄력 개선 효과)

  • Kim, Soo Hyun;Lee, AhReum;Kim, SuJi;Kim, Kyeong Jo;Kwon, Ojun;Choi, Joon Young;Koo, Jin Suk;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.32 no.4
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    • pp.17-24
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    • 2017
  • Objectives : Advanced glycation end products (AGEs) is bind formation of glucose and protein. Acceleration of AGE formation during hyperglycemia is associated with the pathogenesis of diabetic complications and causes kidney and skin damage. The aim of this study was investigated the AGEs inhibitory activity and antioxidant activity of water extracts from young persimmon (YP) and heated young persimmon (HYP). Methods : Paeoniae Radix Alba (YP) is prepared by heating with 30% ethanol. AGEs formation inhibitory activities of YP and HYP measured using bovine serum albumin. To evaluate the protective effects of YP and HYP in diabetic rats induced with streptozotocin (STZ) and methyl glyoxal (MGO), SD rats were distributed into four groups; normal mice (Nor), AGEs-induced rats (Con), AGEs-induced rats treated with 100 mg/kg YP (YP), AGEs-induced rats treated with 100 mg/kg heated YP (HYP) for 3weeks. Heated young persimmon respectively decrease AGEs construction. Results : YP and HYP administration inhibited the biomarkers of AGEs in serum, kidney and skin tissues. AGE-induced rats revealed that the significant decreased collagen however, heat processing methods of young persimmon up regulated inhibits AGEs-induced collagen decrease. The expressions of AGEs were decreased in YP and HYP treated group compared with the control group in tissues. It specifies that HYP has potential to serve as a positive regulator of via AGEs path way. Conclusion : It has proposed that may have an improvement effect on diabetic complications, heated young persimmon has AGEs inhibitory excellent activities and antioxidant effect.

PET Imaging of Click-engineered PSMA-targeting Immune Cells in Normal Mice

  • Hye Won Kim;Won Chang Lee;In Ho Song;Hyun Soo Park;Sang Eun Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.8 no.2
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    • pp.53-61
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    • 2022
  • This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N3) using Ac4ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac4ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [89Zr]Zr(oxinate)4/5 × 106 cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac4ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac4ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [89Zr]Zr(oxinate)4 could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.

Liver Protective Effect of the Co-treatment of Rhei Radix et Rhizoma and Silymarin on TAA-induced Liver Injury (대황과 실리마린의 병용투여의 간섬유화 보호 효과)

  • Il-ha Jeong;Sang-woo Ji;Seong-soo Roh
    • The Journal of Internal Korean Medicine
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    • v.44 no.3
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    • pp.402-417
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    • 2023
  • 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.

The Effect of Baekhogainsam-tang on Metabolism through Modulation of the Gut Microbiota and Gene Expression in High-Fat Diet Induced Metabolic Syndrome Animal Model (고지방식이로 유도된 대사증후군 모델 동물에서 백호가인삼탕(白虎加人參湯)의 장내미생물 및 유전자 발현 조절을 통한 대사 개선 효과)

  • Min-Jin Cho;Song-Yi Han;Soo Kyoung Lim;Eun-Ji Song;Young-Do Nam;Hojun Kim
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.3
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    • pp.1-15
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    • 2023
  • Objectives We aimed to find out the improvement effect of Baekhogainsam-tang (Baihu Jia Renshen-tang, BIT) on metabolic syndrome and alteration of microbiota and gene expression. Methods We used male C57BI/6 mice and randomly assigned them into three groups. Normal control group was fed 10% kcal% fat diet, high-fat diet (HFD) group was fed 45% kcal% fat diet and 10% fructose water. BIT group was fed same diet as HFD group and treated by BIT for once daily, 6 days per week, total 8 weeks. We measured their body weight and food intake every week and performed oral glucose tolerance test 1 week before the end of the study. Then we collected the blood sample to measure triglyceride, total cholesterol, high-density lipoprotein cholesterol, insulin, and hemoglobin A1c. We harvested tissue of liver, muscle, fat, and large intestine for quantitative polymerase chain reaction (qPCR) and histopathological examination. Fresh fecal samples were collected from each animal to verify alterations of gut microbiota and we used RNA from liver tissue for microarray analysis. Results The body weight and fat weight of BIT group were reduced compared to HFD group. The qPCR markers usually up-regulated in metabolic syndrome were decreased in BIT group. Bacteroides were higher in BIT group than other groups. There were also differences in gene expressions between two groups such as Cyp3a11 and Scd1. Conclusions We could find out BIT can ameliorate metabolic syndrome and suggest its effect is related to gut microbiota composition and gene expression pattern.

$V_H$ Gene Expression and its Regulation on Several Different B Cell Population by using in situ Hybridization technique

  • Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.111-122
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    • 1993
  • The mechanism by which $V_H$ region gene segments is selected in B lymphocyte is not known. Moreover, evidence for both random and nonrandom expression of $V_H$ genes in matured B cells has been presented previously. In this report, the technique of in situ hybridization allowed us to analyze expressed $V_H$ gene families in normal B lymphocyte at the single cell level. The analysis of normal B cells in this study eliminated any posssible bias resulting from transformation protocols used previously and minimized limitation associated with sampling size. Therefore, an accurate measure of the functional and expressed $V_H$ gene repertoire in B lymphocyte could be made. One of the most important controls for the optimization of in situ hybridization is to establish probe concentration and washing stringency due to the degree of nucleotide sequence similarlity between different families which in some cases can be as high as 70%. When the radioactive $C{\mu}$ and $V_{H}J558$ RNA probes are tested on LPS-stimulated adult spleen cells, $2{\sim}4{\times}106cpm$/slide shows low background and reasonable frequency of specific positive cells. For the washing condition. 40~50% formamide at $54^{\circ}C$ is found to be optimum for the $C{\mu}$. $V_{H}S107$ and $V_{H}J558$ probes. The analyzed results clearly demonstrate that the level of each different $V_H$ gene family expression is dependent upon the complexity or size of that family. These findings are also extended to the level of $V_H$ gene family expression in separated bone marrow B cells depend upon the various stage of differentiation and conclude no preferential utilization of specific $V_H$ gene family. Thus, the utilization of VH gene segments in B lymphocyte of adult BALB/c mice is random and is not regulated or changed during the differentiation of B cells.

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Effect of Dietary Intake of Salicornia herbacea L. Hot Water Extract on Anti-obesity in Diet-induced Obese Rats (함초 열수추출물의 섭취가 비만유도 흰쥐의 항비만 효과에 미치는 영향)

  • Lee, Hye-Sung;Choi, Jun-Hyeok;Kim, Youn-Geon;Lee, Chang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.7
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    • pp.950-956
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    • 2012
  • Salicornia herbacea L. is an annual herb that grows in salt marshes and salt fields along the seashores. It is also commonly used as a folk remedy in Korea. The objective of this study was to investigate the beneficial effects of Salicorrnia herbacea L. (SH) hot water extract on obesity. Five-week old male Sprague-Dawley rats (n=36) were divided into two groups and provided either a normal fat diet (11.5% fat from kcal) or a high fat diet (40.5% fat from kcal) for 6 weeks to induce obesity. Then, rats were blocked into six groups of six mice each and provided either a diet containing SH (0.5% of diet; g/kg) or a normal diet for another 6 weeks. Final body weights were significantly reduced when rats were fed SH among the high fat diet groups (HNS and HHS). Serum triglyceride concentrations significantly decreased in every group provided SH. HDL-cholesterol concentrations significantly increased in SH-fed groups among the high fat diet groups. Further, atherogenic index significantly decreased when rats were fed SH diet (HHS). There were no differences in LDL-cholesterol between the high fat diet groups, and glucose concentrations decreased when rats were fed SH diet (HNS). These results indicate that dietary intake of Salicornia herbacea L. hot water extract might have beneficial effects on obesity by reducing body weight, fat weights, and improving blood lipid profile.

Effects of Bacillus Calmette-Guerin (BCG) on the DNA Synthesis of Mouse Duodenal Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells (Bacillus Calmette-Guerin (BCG) 투여가 Ehrlich 종양세포를 이식한 생쥐 샘창자 상피세포의 DNA합성에 미치는 영향)

  • Ko, Jeong-Sik;Kim, Heung-No;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Duk-Soo
    • Applied Microscopy
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    • v.40 no.4
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    • pp.193-200
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    • 2010
  • This experiment was performed to evaluate the morphological responses of the duodenal epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Bacillus Calmette-Guerin (BCG). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8\sim0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of $0.7{\mu}Ci$/g of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and duodenal tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab, England) in a dark room and dried and were placed in a light-tight box. The number of labeled epithelial cells in the duodenal mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On the light microscopic study, duodenal mucosae had no severe morphological changes following the injection of BCG. In the tumor control and BCG treated groups, a number of small lymphocytes and eosinophile leucocytes are slightly increased as compared with those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 632.3 (${\pm}14.47$), 761.7 (${\pm}27.65$) and 505.0 (${\pm}17.09$) respectively. From the above results, BCG may suppress the DNA synthesis of the duodenal epithelial cells, but does not results severe structural defect on the duodenal mucosae. And it is suggested that BCG may greatly improve the anticancer therapy on certain kind of cancer.

Effect of Tumor Hypoxia on Efficacy of Tirapazamine Combined with Fractionated Irradiation in Mouse Tumor (마우스종양에서 분할방사선조사와 병용된 Tirapazamine의 효과에 미치는 종양 저산소상태의 영향)

  • Kim, Il-Han
    • Radiation Oncology Journal
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    • v.18 no.2
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    • pp.120-126
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    • 2000
  • Purpose : Tumor hypoxia can be overcome with hypoxic cytotoxin. In mouse tumor, tirapazamine's efficacy of the potentiating radiation effect was tested by the tumor oxygenation status combined with hype facti on ated rad iotherapy .:The control and hypoxic mouse tumors we established by inoculation of RIF-1 tumor cells into the normal or previously irradiated back and thigh of C3H mice. When the tumors reached a proper size, both the control and hypoxic tumors were given hypefractionated treatments (8fractions/4 days) with saline (0.02 ml/g), tirapazamin (0.08 mM/0.02 ml/kg), irradiation (2.5 Gy), irradiation combined with tirapazamine given 30 minutes prior to each irradiation. The response was evaluated by the growth delay assay by measuring tumor size from day 0 (12 hrs prior to the first fractionation) to the day when the volume had 4-fold increase or cross sectional area had 2-fold increase. Results : Overall growth pattern showed that tirapazamine Potentiated radiation effect in back and thigh tumors grew in the normal and preirradiated tumor bed. With growth delay assay using reference point of initial tumor volume or cross sectional area, tirapazamine potentiated radiation effect 1.9 times for the control and 2.4 times for the hypoxic tumors in back, and 1.85 times for the control and 1.6 times for the hypoxic tumors. With reference of 4-fold increase of the initial volume or 2-fold increase of the cross sectional area, tirapazamine potentiated radiation effect 1.48 times for the control and 2.02 times for the hypxic tumors in back, and 1.85 times for the control and 1.6 times for the hypoxic tumors. Conclusions : Present result indicated that radiation response of hypoxic tumors was potentiated by tirapazamine in the back or thigh tumors grew in the control or preirradiated tumor bed, and potentiation of the hypoxic tumors was eDual to or greater than that of the control tumors in the back or thigh.

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Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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Antibacterial Activity of Ceftizoxime Against Gram Negative Enteric Bacteria in vitro and in vivo (Ceftizoxime의 장내세균에 대한 시험관내 및 생체내 항균효과)

  • Byun, Woo-Mok;Chang, Jae-Chun;Park, Bok-Hwan;Kim, Hee-Sun;Kim, Sung-Kwang
    • Journal of Yeungnam Medical Science
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    • v.6 no.1
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    • pp.59-68
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    • 1989
  • Ceftizoxime sodium is a new synthetic ${\beta}$-lactam antibiotic combining potent antibacterial activity with high stability to a wide range of bacterial ${\beta}$-lactamase. This experiment was achieved to evaluate the antibacterial activities of ceftizoxime sodium againist Gram negative enteric bacteria isolated from in outpatient visiting Yeungnam university hospital and to study the emergence of drug induced bacterial varients which resist to ceftizoxime in vitro. The antibacterial activity of the ceftizoxime was compared with that of antibiotics and its effect on population of normal intestinal flora in mice was observed. The results are summarized as follows : 1. Highly effective antibacterial activity of ceftizoxime against Gram negative enteric bacilli was demonstrated and this antibacterial activity was superior to that of ampicillin. 2. Several test strains shows multiple antibiotic resistence. Among 15 strains of Escherichia coli, 1 strain was resistent to ampicillin, cefadroxyl, gentamicin, tetracycline, and 2 strains were resistent to ampicillin, cefadroxyl, tetracycline, five strains of Escherichia coli and Enterobacter cloacae was resistent to amplicillin, tetracycline and Shigella dysenteria was resistent to ampicillin, gentamicin, tetracycline. 3. The frequency of in vitro emergence of resistent varients among ceftizoxime sensitive bacteria in the presence of increasing concentrations of the compound was found to be low. 4. Plasmid was isolated in 6 of 9 strains (6 strains of Escherichia coli, Shigella dysenteriae, Enterobacter cloaceae and Salmonella typhi) That showed different antibiotic resistance. They were 5 strains of Escherichia coli and 1 strain of Shigella dysenteriae. However, plasmid could not be considered as a hallmark for antibiotic resistance by this. Further studies with curing experiment are to be accomplished for this purpose. 5. Changes in the bacterial count of normal intestinal flora following 25mg/kg/day administration of ceftizoxime over S consecutive days were not significant. In conclusion, ceftizoxime appeared to be a drug of choice in the treatment of Gram negative enteric bacilli infection.

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