• The Journal of Pediatrics of Korean Medicine
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• v.12 no.1
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• pp.211-230
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• 1998

Introduction The effects of Bojungikkitang on the immunosuppression induced by methotrexate in rats were investigated in this experiment. The multiple parameters of immunity assessed in. each rats includes the rate of body weight loss, weight changes in thymus, spleen and axillary lymphnode. The number of lymphocyte and CD4+ T cell count in blood, thymus, spleen and axillary lymphnode were also measured. Methodology Male Sprague-Dawley rats were chosen as an experiment object and were divided into 3 groups by a random selection. Each group consisted 6 rats. The normal group didn't receive any treatment. The control group was administered methotrexate for 4 days. The sample group was administered with both Bojungikkitang and methotrexate for 4 days. The dosage of medication was 2cc/day, 1cc given at 10AM and another 1cc given at 5PM. Results The rate of body weight loss was significantly decreased in the sample group. The weight of thymus was significantly increased in the sample group while the weight of spleen did not show much increase. Blood CD4+ T cell count, thymus lymphocyte count, thymus CD4+ T cell count, spleen lymphocyte count, spleen CD4+ T cell count and axillary lymph node CD4+ T cell count were significantly increased in the sample group while blood lymphocyte count and axillary lymphnode lymphocyte count did not show much increase. Conclusion As one can witness from the above results, administration of Bojungikkitang played potent role in increasing immune system among the rats treated with methotrexate which induces immunosuppression. Overall increase of lymphocyte count and CD4+ T cell count in the sample group with Bojungikkitang effectively proves its ability to boost the immune system.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.2
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• pp.263-272
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• 2007

This experiment was conducted to examine the effects of cage type (CT) and cage density (CD) on physiological variables in growing rats. Sprague Dawley rats (n=108) weighing an average of 46 g were housed in metallic cage with woodchip bedding (MCWB), metallic cage with wire mesh (MCWM), and plastic shoebox with woodchip bedding (PCWB) separately by sex at normal ($160-cm^2/rat$, ND) and high ($80-cm^2/rat$, HD) CD from 3 to 10 wks of age. All cages were in dimension of $24{\times}40{\times}20$ cm ($W{\times}D{\times}H$). At the end of the experiment, blood samples were collected and 6 rats from each cage were sacrificed. No death was observed among rats at ND, whereas mortality rate at HD was 22.3% for males and 13.9% for females. Heart weight was affected by CT. Doubling CD caused 23, 11.8, 17.9, 8.6, 6.9, and 16.4% decreases in BW and weights of heart, liver, kidney, testis, and ovary, respectively. Except for adrenal gland, other organs for males were heavier than for females. Liver weight of males and females responded differently to CT and CD. Comparing with females, males had 7.3 and 5.2% heavier and 9.9% lighter liver weights in MCWB, MCWM, and PCWB, respectively. As CD doubled, liver weight for males and females decreased by 22.4 and 13.1%, respectively. Mean adrenal gland weight increased by 8.4% and decreased by 9.7% for males and females, respectively, with doubling CD. CT affected glucose, TG, Ca, and ALP levels. However, CD did not alter blood chemistry. Rats housed in metallic cages had greater neutrophil count and neutrophil:lymphocyte ratio than rats housed in plastic cages. Doubling CD caused a 24.2% increase in lymphocyte count. There were CT by CD, CT by sex, and CD by sex interaction effects on lymphocyte count. Doubling CD caused 0.1% decrease and 49.8 and 26.7% increases in lymphocyte count for rats housed in MCWB, MCWM, and PCWB, respectively. Comparing with females, lymphocyte count for males housed in MCWB, MCWM, and PCWB had 8.9 and 12.9% greater and 30.3% less lymphocyte counts, respectively. Lymphocyte count decreased by 4.12% for males, whereas it increased by 61.0% for females as CD doubled. Doubling CD resulted in 2.5 and 2.3% increases in erythrocyte count and hematocrit value. These data suggest that animals perform better in metallic cages than in plastic cages and that cage density had pronounceable effects on physiological parameters in a cage type and sex dependent-manner.

• The Journal of Pediatrics of Korean Medicine
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• v.12 no.1
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• pp.257-276
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• 1998

Introduction The effects of Samchulkunbitang on the immunosuppression induced by methotrexate in rats were investigated in this experument. The multiple parameters of immunity assessed in each rats includes the rate of body weight loss, weight changes in thymus, spleen and axillary lymphnode. The number of lymphocytes and CD4+ T cell count in the blood, thymus, spleen and axillary lymphnode were also measured. Methodology Male Sprague-Dawley rats were chosen as an experiment object and were divided into 4 groups by a random selection. Each group consisted of 6 rats. The normal group didn't receive any treatment. The control group was administered with methotrexate for 5 days. The sample Ⅰ group was administered with Samchulkunbitang for 5 days. The sample Ⅱ group was administered with both Samchulkunbitang and methotrexate for 5 days. The dosage of medication was 2cc/day, 1cc given at 10 AM and another 1cc given at 5 PM. Results The rate of body weight loss was significantly increased in the Sample Ⅰ and Sample Ⅱ group. The weight of thymus and spleen were significantly increased in Sample Ⅰ and Sample Ⅱ groups While the weight of axillary lymphnode did not show much increase. No significant differences were measured among the experimental groups. Blood lymphocyte count, blood CD4+ T cell count, spleen lymphocyte count, axillary lymphnode lymphocyte and CD4+ T cell count were significantly increased in Sample Ⅰ and Sample Ⅱ groups while spleen CD4+ T cell count did not show much increase. No significant differences were measured among the experimental groups. Conclusion As one can witness from the above results, administration of Samchulkunbitang played potent role in increasing immune system among the rats treated with methotrexate which induces immunosuppression. Overall increase of lymphocyte count and CD4+ T cell count in the sample groups with Samchulkunbitang effectively proves its ability to boost the immune system.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.111-129
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• 1998

In order to research the effect of Sipjundaebotang on the immune respons of CD4+ T cell and CD8+ T cell in rats, author have performed this experimental study. Experimental groups are divided into 5 groups(intact, control, sample I, sample II, sample III). Normal group was intact, control group was administrated normal saline 2cc for 5 days, sample I was administrated low concentration of Sipjundaebotang(100/3100pack/ml) 2cc for 5 days, sample II was administrated middle concentration of Sipjundaebotang(500/3100pack/ml) 2cc for 5 days, sample III was administrated high concentration of Sipjundaebotang(2500/3100pack/ml) 2cc for 5 days. WBC, Lymphocyte, CD4+ T cell in the blood, spleen, axillary node and CD8+ T cell in the blood, spleen and axillary node were determined. The results was as follows: 1. WBC count in blood was significantly decrcased in the control, sample I, II, III groups as compared with the normal group. 2. Lymhocyte count in the blood was significantly decreased in the control, sample I, II groups as compared with the normal group. 3. CD4+ T cell count in the blood was significantly increased dose-dependently in the sample I, II, III groups as compared with the normal group. 4. CD4+ T cell count in the spleen was significantly increased in the sample I, II, III groups as compared with the normal group. 5. CD4+ T cell count in the axillary node was significantly increased in the sample III group as compared with the normal group, however significantly decreased in the sample I, II groups as compared with the normal group. 6. CD8+ T cell count in the blood was significantly increased in the sample III groups as compared with the normal group. 7. CD8+ T cell count in the spleen was significantly increased in the sample I, II, III groups as compared with the normal group. but there was no differences in the sample groups. 8. CD8+ T cell count in the axillary node was decreased in the sample I, II groups as compared with the normal group.

• IMMUNE NETWORK
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• v.14 no.6
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• pp.289-295
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• 2014

Flow cytometric immunophenotyping of peripheral blood lymphocyte subsets is a powerful tool for evaluating cellular immunity and monitoring immune-mediated diseases. The numbers and proportions of blood lymphocyte subsets are influenced by factors such as gender, age, ethnicity, and lifestyle. This study aimed to establish reference ranges for peripheral blood lymphocyte subsets in a healthy Korean population. Blood samples from 294 healthy adults were collected. Lymphocyte subsets were analyzed using a single-platform method with a flow cytometer; white blood cells and lymphocytes were analyzed using an automated hematology analyzer. The mean value of the white blood cell count was $5,665cells/{\mu}l$, and the mean values of the subtype counts (percentages) were as follows: lymphocytes, $1,928cells/{\mu}l$ (35.08%); $CD3^+$ cells, $1,305cells/{\mu}l$ (67.53%); $CD3^+CD4^+$ cells, $787cells/{\mu}l$ (40.55%); $CD3^+CD8^+$ cells, $479cells/{\mu}l$ (25.23%); $CD3^-CD19^+$ cells, $203cells/{\mu}l$ (10.43%); and $CD3^-CD56^+$ cells, $300cells/{\mu}l$ (15.63%). Additionally, the $CD4^+/CD8^+$ ratio was 1.81. In this study, gender and age significantly influenced blood lymphocyte subsets. Our results demonstrate that, as with other populations, a healthy Korean population has its own, region-specific, lymphocyte subset reference ranges.

• Korean Journal of Veterinary Research
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• v.50 no.3
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• pp.179-185
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• 2010

The present study was undertaken to establish reference values for the composition blood lymphocyte populations and compare forty three Hanwoo neonatal calves (KC) with twenty one Holstein calves (HC) by blood cell count and immunophynotying. The percentages of CD2+, CD4+, CD8+, CD26+, ACT2+, MHC class, MHC class II and WC1+ T cells, B cells were determined by flow cytometry. The number of lymphocyte and monocyte in HC were higher than those of KC. However, the number of neutrophils was higher in HC than KC. The proportions of CD2+, CD4+, CD8+, MHC class, and WC1+ lymphocytes remained relatively stable during the study period, while there was a moderate increase in the relative percentage of CD26+, ACT2+, MHC class II and B cell from birth to approximately 3 weeks of age. Marked differences in the relative proportions of the lymphocyte subpopulations were noted between the individual calves. The present study shows that the T-cell subpopulations are present in peripheral blood of KC at levels comparable with HC, while the MHC class II and B cell population of KC increases significantly with age. The absolute number of WBC in KC was due to the decrease of absolute number of neutrophil rather than the increase of lymphocyte. The results indicated that KC have significantly higher number of neutrophils, and proportion of MHC class II and B cell than HC.

• IMMUNE NETWORK
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• v.1 no.3
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• pp.236-243
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• 2001

Background: An immunological approach for aging mechanism appears to be important. Lymphocyte subsets analysis in peripheral blood is widely performed to assess the immune status and to diagnose and monitor various diseases. Some lymphocyte subsets are known to change with age, but only few data about age-related reference ragnes for these subsets in healthy individuals have been reported. So we attempted to report reference ranges for these subsets in each age group and review changes of the results with age for the secondary studies about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement (VDJ) including recombination activating genes (RAG-1 and RAG-2). Methods: Lymphocyte subset analysis was performed on 302 subjects, 189 males and 113 females with age group of all decades of life. Two color direct immunofluorescene flow cytometry (FCM) was done using $Simultest^{TM}$ IMK-Lymphocyte kit (Becton Dickinson, USA), $FACScan^{TM}$ (Becton Dickinson, USA) and $FACSCalibur^{TM}$ (Becton Dickinson, USA). Lymphocyte subsets analysed were T ($CD3^+$) and B cells ($CD19^+$), helper/inducer T ($CD4^+$) and suppressor/cytotoxic T cells ($CD8^+$), helper/suppressor ($CD4^+/CD8^+$) ratio and natural killer (NK) cells ($CD3^-CD16^+/CD56^+$). The absolute numbers of each subset were calculated from total lymphocyte counts. Data collected was analysed using SAS 6.12. A P-value of < 0.05 was considered significant. Results: We reported the counts and percentages of lymphocyte and these subsets in each age group. There were no statistically significant differences between male and female subjects. The percentage of $CD4^+$ T cells, and the count of NK cells did not show the significant difference among the various age groups. The age-related changes observed in our study were as following: 1) a decrease in the percentages of T cells, B cells and $CD8^+$ T cells ; 2) a decrease in the counts of B cells and $CD8^+$ T cells ; 3) an increase in the percentage and count of NK cells ; and 4) an increase in the $CD4^+/CD8^+$ ratio. Conclusion: The characteristics of aging process appeared to be showing a marked decrease of lympocyte subsets T and B cells as well as T8 ($CD8^+$). The age-related increase of the percentage of cells bearing NK marker can be interpreted as a compensatory consequence to cope with the decrease of T cells related to the thymic involution. These changes with age appeared to be for the secondary study about immune cell function as lymphocyte blast transformation and immunoglobulin gene rearrangement.

• Radiation Oncology Journal
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• v.14 no.1
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• pp.53-59
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• 1996

Purpose : Radiation-induced alteration in the immune function is well known phenomenon in cancer patients. Our purpose is to evaluate the extent of immune suppression immediately after mediastinal or pelvic irradiation, which include significant volume of active bone marrow in adults. Materials and Methods'48 cancer patients with mediastinal(N=29) and pelvic irradiation(N=19) were the basis of this analysis. Age ranged from 36 to 76 and mean and median value was 57 years, respectively Sex ratio was 1.3(M: F=27/21). The immunological parameters were the complete blood cell(CBC) with differential cell(D/C) count, T cell subset(CD3, CD4, CD8 CDl9), NK cell test(CDl6, CD56), and serum immunoglobulin(IgG, IgA, IgM) level. Results : The mean value of white blood cell(WBC) was reduced from 7017 to 4470 after irradiation(p=0.0000). In the differential count, the number of lymphocyte, neutrophil, and basophil was markedly reduced with statistical significance(p<0.01) and the number of monocyte was not changed and, on the contrary, that of eosinophil was increased by irradiation. In the lymphocyte subpopulation analysis, the number of all subpopulations, CD3(T cell), CD4(helper T cell), CD8(suppressor T cell), CDl6(NK cell), CDl9(B, cell) was reduced with statistical significance. The mean ratio of CD4 to CD8 in all patients was 1.09 initially and reduced to 0.99 after radiotherapy(p=0.34) , but the proportional percentage of all subpopulations was not changed except CD19(B cell) after irradiation. In the immunoglobulin study, initial values of Ig G, Ig A, and Ig M were relatively above the normal range and the only Ig M was statistically significantly reduced after radiotherapy(p=0.02). Conclusion : Mediastinal and pelvic irradiation resulted in remarkable suppression of lymphocyte count in contrast to the relatively good preservation of other components of white blood cells. But the further study on the functional changes of lymphocyte after radiotherapy may be necessary to conclude the effects of the radiation on the immunity of the cancer Patients.

• The Journal of Internal Korean Medicine
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• v.25 no.4
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• pp.117-128
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• 2004

Objectives : To examine the effects of Gojineumja on white rats which deteriorated immunity caused by Methotrexate(MTX), first of all, MTX was fed to the rats once a day for 4 day. Methods : After the immune response of the rats are deteriorated, dried extracts of Gojineumja(GJE) mixed in water was fed to the white rats once a day for l4days. The next conclusion was made by examining the rates of B-cells and T-cells of the peripheral blood and the changes in rates of CD4+ T-cells and CD8+ T -cells of the blood sampled from the spleen and peripheral region. Especially the count of CD3+ CD4+ T-cells of the peripheral blood and the count of CD3+ CD4+ T-cells of the spleen the count of CD4+/ CD8+ T-cell of the peripheral blood and the spleen proved the significant effect of increasing immune responses statistically. Results :(1) The following are the summary of the results. (2) The percentage of B lymphocyte of peripheral blood was increased significantly in GJE group as compared with control group. (3) The percentage of CD3+ CD4+ T-cell of peripheral blood was increased significantly in GJE group as compared with control group. (4) The percentage of CD3+ CD8+ T-cell of peripheral blood was not different statistically. (5) The percentage of CD4+/ CD8+ T-cell of peripheral blood was increased significantly in GJE group as compared with control group. (6) The percentage of CD3+ CD4+ T-cell of spleen was increased significantly in GJE group as compared with control group. (7) The percentage of CD3+ CD8+ T-cell was not different statistically. (8) The percentage of CD4+ /CD8+ T-cell was not different statistically. Conclusions : Gojineumja has an effect of increasing immune responses on white rats with deteriorated immunity caused by MTX.

• Korean Journal of Veterinary Pathology
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• v.1 no.1
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• pp.1-6
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• 1997

The cellularity and composition of the spleen lymph node thymus and peripheral blood and tempo of regeneration were studied at various time points after syngeneic bone marrow transplantation(BMT) in C3H/Hen mice. Significant depression of absolute lymphocyte count was noted on week 1 after lethal whole-body irradiation and BMT. In comparison to the lymph node thymus and spleen had an rapid regeneration of cellularity. The distinct cell populations($CD4^+,\;CD8^+,\;CD28^+,\;B220^+) have determined in the lymphoid tissue of mice subjected to irradiation. The relative representation of these subpopulations was significantly different from that in nonirradiated control. $CD4^+\;and\;CD8^+$ cells were present in very low numbers whereas the $B220^+$ cells reached more than normal range at 2 weeks after BMT. The number of $CD4^+$ cells returned to normal relatively soon than $CD8^+$ cell. At week 4 after BMT, the cellularity and composition of spleen lymph node and peripheral blood lymphocyte reached about 50% of the normal range therefore we can choose this time point for the other tests of immune function after BMT.