• Korean Journal of Acupuncture
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• 제39권4호
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• pp.132-141
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• 2022

Objectives : Some acupoints are commonly utilized to treat a variety of diseases. The acupoints appear to have a wide range of effects caused by several mechanisms. The purpose of this study is to investigate into the potential role of microRNAs (miRNAs) in the multipotent effects of individual acupoint stimulation. Methods : We examined the miRNA expressions in the dorsal root ganglia (DRG) of neuropathic or inflammatory pain rats following ST36 and GB34 electroacupuncture (EA) stimulation. Neuropathic pain was induced by L5 spinal nerve ligation. Inflammatory pain was induced by knee joint injection of Complete Freund's adjuvant (CFA). EA was given under gaseous anesthesia with the same parameters (1mA, 2Hz, 30 min) in 5 consecutive days. Pain behaviors and miRNA expressions were analyzed. Results : In rats with neuropathic and inflammatory pain, EA treatments significantly enhanced the paw withdrawal threshold and weight-bearing force. After nerve injury, 36 miRNAs were upregulated in the DRG of neuropathic rats, while EA downregulated 10 of them. Furthermore, 14 miRNAs were downregulated following nerve damage, while one was increased by EA. 15 miRNAs were increased in the DRG of inflammatory rats following CFA injection, while 5 were downregulated by EA. Furthermore, 17 miRNAs were downregulated following CFA injection, while 7 were increased by EA. The miRNAs rno-miR-335, rno-miR-381-5p, rno-miR-1306-3p, and rno-miR-1839-3p were regulated by EA in both models. Conclusions : In two pain models, EA applied to ST36 and GB34 regulated miRNA expression differently. There appeared to be both acupoint-specific and non-specific miRNAs, and miRNA regulation of differential protein expression may modulate a variety of EA mechanisms.

• 대한약침학회지
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• 제26권3호
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• pp.257-264
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• 2023

Objectives: The study was designed to evaluate anti-arthritic activity of Ajmodadi Churna (AC) and its effect on Complete freund's adjuvant (CFA)-induced arthritis in Wistar rats. Methods: Arthritis was induced by injecting 0.2 mL CFA into sub plantar surface of left hind paw. Test sample AC-1 and AC-2, 200 and 400 mg/kg, respectively was given to the animals for 21 consecutive days. The increase in swelling was observed after induction of arthritis. The paw edema was measured on 0, 3, 7, 14 and 21 day using Vernier caliper after the induction of arthritis. The collected blood samples further used for the estimation of red blood cells (RBC), white blood cells (WBC), erythrocytes sedimentation rate (ESR), and hemoglobin (Hb), using hematology analyzer. Serum concentration of IL-6 and TNF-α were also measured using rat ELISA kits. Results: Results showed that a significant reduction in paw edema was observed in AC-2 treated rats. The paw edema was restored on day 21 was 4.48 mm for AC-2, which is near to the control group. The arthritis score in treated rats was found to be considerably lower than in the control group i.e. 0.83 for AC-2 and 1.50 for AC-1. A decrease in levels of RBC and hemoglobin were observed in arthritic rats. Inflammation was significantly reduced and serum levels of IL-6 and TNF-α were lowered after treatment with the test drug. Conclusion: It can be concluded from the study that AC possess significant anti-arthritic activity. Furthermore, this condition was linked to a reduction in abnormal humoral immune responses.

• IMMUNE NETWORK
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• 제20권4호
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• pp.33.1-33.19
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• 2020

We tested how adjuvants effect in a cancer vaccine model using an epitope derived from an autoactivation loop of membrane-type protease serine protease 14 (PRSS14; loop metavaccine) in mouse mammary tumor virus (MMTV)-polyoma middle tumor-antigen (PyMT) system and in 2 other orthotopic mouse systems. Earlier, we reported that loop metavaccine effectively prevented progression and metastasis regardless of adjuvant types and TH types of hosts in tail-vein injection systems. However, the loop metavaccine with Freund's complete adjuvant (CFA) reduced cancer progression and metastasis while that with alum, to our surprise, were adversely affected in 3 tumor bearing mouse models. The amounts of loop peptide specific antibodies inversely correlated with tumor burden and metastasis, meanwhile both T_H1 and T_H2 isotypes were present regardless of host type and adjuvant. Tumor infiltrating myeloid cells such as eosinophil, monocyte, and neutrophil were asymmetrically distributed among 2 adjuvant groups with loop metavaccine. Systemic expression profiling using the lymph nodes of the differentially immunized MMTV-PyMT mouse revealed that adjuvant types, as well as loop metavaccine can change the immune signatures. Specifically, loop metavaccine itself induces T_H2 and T_H17 responses but reduces T_H1 and T_reg responses regardless of adjuvant type, whereas CFA but not alum increased follicular T_H response. Among the myeloid signatures, eosinophil was most distinct between CFA and alum. Survival analysis of breast cancer patients showed that eosinophil chemokines can be useful prognostic factors in PRSS14 positive patients. Based on these observations, we concluded that multiple immune parameters are to be considered when applying a vaccine strategy to cancer patients.

• Journal of Microbiology and Biotechnology
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• 제34권7호
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• pp.1419-1424
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• 2024

Secretin PilQ is an antigenically conserved outer membrane protein that is present in most meningococci and PorA is a major protein that elicits bactericidal immune response in humans following natural disease and immunization. In the present study, BALB/c mice were immunized subcutaneously with rPilQ406-770 or rPorA together with Freund's adjuvant (FA). Serum antibody responses to serogroup A and B Neisseria meningitides whole cells or purified proteins and functional activity of antibodies were determined by ELISA and serum bactericidal assay (SBA), respectively. Serum IgG responses were significantly increased in the immunized group with rPilQ406-770 or rPorA together with FA compared to control groups. IgG antibody response of mice immunized with rPilQ406-770 was significantly more than mice immunized with rPorA (OD at 450 nm was 1.6 versus 0.83). The booster injections were effective in increasing the responses of antirPilQ406-770 or anti-rPorA IgG significantly. Antisera produced against rPilQ406-770 or rPorA demonstrated strong surface reactivity to serogroup B N. meningitides in comparison with control groups. Antisera raised against rPorA or rPilQ406-770 and FA demonstrated SBA titers from 1/1024 to 1/2048 against serogroup B. The strongest bactericidal activity was detected in sera from mice immunized with rPilQ406-770 mixed with FA. These results suggest that rPilQ406-770 is a potential vaccine candidate for serogroup B N. meningitidis.

• 한국미생물·생명공학회지
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• 제24권1호
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• pp.119-125
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• 1996

In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

• 한국전문물리치료학회지
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• 제13권3호
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• pp.33-40
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• 2006

The purpose of this study was to assess the effects of High-Voltage Pulsed Current (HVPC) and ultrasound on adjuvant-induced arthritis in rats. Adjuvant arthritis was induced in female Sprauge-Dawley rats by the subcutaneous injection of a single dose of $.1m{\ell}$ of Complete Freund's Adjuvant (CFA) (1 mg of Mycobacterium Butyricum suspended in $.1m{\ell}$ paraffin oil) into the right hind paw. A randomized, parallel-groups design of 24 subjects was used. All rats were randomly assigned to control (n=8), ultrasound (n=8), and HVPC (n=8) were compared with those of injured rats. The rats in the pulsed ultrasound group were treated at 1 MHz frequency with $.5W/cm^2$ intensity in 1:4 mode for 5 minutes per day. The rats in the HVPC group were treated at 120 pulses per second and $50{\mu}s$ phase duration, 20 mA intensity for 30 min per day. Treatment was done in the left and right hind limb for 2 weeks. We evaluated clinical, radiographic, hematologic and histopathologic findings before and after treatment and obtained the following results. 1. Edema of the right hind paw was more significantly reduced in the ultrasound and HVPC groups than the control group on days 9, 12, and 14 (p<.05). Edema of the left hind paw was more significantly reduced in ultrasound and HVPC groups than the control group on days 12, 14 (p<.05). 2. WBC counts of the ultrasound and HVPC groups as compared with the control group were becoming remarkably decreased after the treatment. 3. In radiologic findings, arthritis formation was seen according to the score of arthritis, which was the highest in the control group, upon the observation of radiographs of the left and right hind paws. However, no statistically significant difference was present in the score within three groups. 4. In the histopathologic findings, ultrasound and HVPC groups had effectively suppressed erosions of articular cartilage and inflammatory cell infiltrations. Therefore, the results of the study show that rats that were treated with the ultrasound and HVPC effectively suppressed adjuvant arthritis. However, no statistically significant difference was present between the ultrasound group and the HVPC group.

• 한국전문물리치료학회지
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• 제10권2호
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• pp.45-59
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• 2003

The purpose of this study was to assess the effects of ultrasound on adjuvant-induced arthritis in rats. Adjuvant arthritis was induced in 26 female Sprauge-Dawley rats by the subcutaneous injection of a single dose of .1 mL of Complete Freund's Adjuvant (CFA) (1 mg of Mycobacterium Butyricum suspended in .1 $m{\ell}$ paraffin oil) into the right hind paw. After confirming inflammatory edema and arthritis in the paw 2 weeks later, the arthritic rats were divided into 3 groups, i.e., a control group, a pulsed ultrasound group (Group A), and a continuous ultrasound group (Group B) with 8 rats placed in each group. The rats in Group A were treated with pulsed ultrasound at 1 MHz frequency with .5 $W/cm^2$ intensity in 1 : 4 mode for 3 minutes. The rats in Group B were treated with continuous ultrasound at 1 MHz frequency with 2 $W/cm^2$ intensity in the continuous mode for 3 minutes. The ultrasound treatment was done in the left and right ankles for 2 weeks. Clinical, radiographic and histopathologic findings were then evaluated before and after treatment and yielded the following results. 1. No significant difference was present in body weight between the control group and the treated groups. 2. A statistically significant decrease in the edema of the paw was seen in the rats in Group A that was treated with pulsed ultrasound by 26~29 days after the treatment started (p<.05). 3. According to radiological examination, Group A showed the lowest score in arthritis scale which means it showed a tendency to suppress arthritic inflammation of the left and right hind paws. However, no statistically significant difference was present in the score between the control group, Group A and Group B. 4. According to histopathologic findings, the degree of infiltration by inflammatory cells and hypertrophy of the synovium were less in Group A compared with the control group and Group B. The results of the study show that rats that were treated with the pulsed ultrasound effectively suppressed adjuvant arthritis. However, more effort is needed to objectively prove the effectiveness of ultrasound by developing more sensitive testing methods that could quantitatively evaluate the treatment effects of acute rheumatoid arthritis and by trying out different ultrasound treatment methods.

• Toxicological Research
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• 제18권1호
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• pp.87-98
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• 2002

This study was conducted to evaluate the safety of a recombinant human Factor VIII(GC-$\gamma$ AHF) manufactured by Korea Green Cross Company with different technology according to the Regulation of Korean Food and Drug Administration (l 998. 12. 3). In acute toxicity test, both genders of Sprague-Dawley rats and Beagle dogs were administered intravenously with GC-$\gamma$ AHF of three doses (3,125, 625 and 125 IU/kg), and single dose of 3,125 IU/kg, respectively. No dead animal and abnormal autopsy findings were found in Control and GC-$\gamma$ AHF treated group. Therefore, the 50% lethal dose ($LD_{50}$) of GC-$\gamma$ AHF was conidered to be higher than 3,125 IU/kg in rats and dogs. In the four weeks repeated intravenous toxicity study, GC-$\gamma$ AHF was administrated intravenosly to both genders of rats and dogs with 3 doses (500, 150, 50 IU/kg). There were neither dead animals nor significant changes of body weights during the experimental Period. In addition, no significant GC-$\gamma$ AHF related changes were found in clinical sign, urinalysis and other finding. Statistically changes were observed in hematological, biochemical and organ weight parameters of treated groups: however these changes were not dose dependent. No histopathological lesion were observed in both control and treated animals. Above data suggest that no observed adverse effect level of test materials in rats and dogs might be over 500 IU/kg/day in this study. In ocular irritation test, any injury on iris, conjunctiva and cornea in rabbits were not observed. The acute ocular irritation index (A.O.I.), mean ocular irritation index (M.O.I.) and Day-7 individual ocular irritation Index (I.O.I.) of GC-$\gamma$ AHF were 0. In the primary skin Irritation test, the primary irritation index (P.I.I.) oj GC-$\gamma$ AHF were 0. Therefore, the GC-$\gamma$ AHF is considered not to have the primary skin and eye toxicity in rabbits. In active systemic anaphylaxis (ASA) test, GC-$\gamma$ AHF and GC-$\gamma$ AHF emulsified with Freund's complete adjuvant (FCA) did not induce any symptom of anaphylactic shock in guinea pigs. In passive cutaneous anaphylxis (PCA) test, after sensitization with antisera of GC-$\gamma$ AHF sensitized mice, blue spots were observed on the hypodermis of back of rats, but diameter of each spot was smaller than 5 mm in each test groups except the positive control group. Based on the results of this study, GC-$\gamma$ AHF is not conidered to have any antigenic potential. In conclusion, at levels of up to 500 IU/kg, GC-$\gamma$ AHF did not produce treatment-related toxicity under the conditions of these acute-, four week repeated-toxicity, primary skin and eye toxicity, and antigenicity test.

• 대한약침학회지
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• 제2권1호
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• pp.93-109
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• 1999

To investigate effects of Achyranthis Radix herbal-acupuncture on adjuvant arthritis in rats, the edema rate, the number of WBC, the quantity of total protein, albumin and globuline in the blood serum and histological test of the muscular tissue were measured in the arthritis part. 1. After elicitating arthritis of Sprague dawley(SD) rats by injection of Freund's complete adjuvant for 2 weeks, normal saline was injected for the Exp. I group and Achyranthis Radix herbal-acupuncture was injected for the Exp. II group during 30days. Selected point was $D\acute{u}b\acute{i}(ST_{35})$ in both the groups. And then the volume of the paw were checked. The volume of the paw was $0.84{\pm}0.14mm$ in the Exp. I group and $0.47{\pm}0.11mm$ in the Exp.II group, the swelling of the paw was restricted significantly in the Exp. II group(p<0.05). 2. The number of WBC was $10.34{\pm}0.14(10^3/ml)$ in the normal group and $37.47{\pm}5.46(10^3/ml)$ in the Exp. I group. It was $21.24{\pm}2.58(10^3/ml)$ in the Exp. II group. This fact showed that the group Exp. II with Achyranthis Radix herbal-acupuncture was more effective than the Exp. II group in the treatment of arthritis(p<0.05). 3. The content of the total protein in the blood serum was $6.14{\pm}0.43g/dl$ in the normal group, $7.95{\pm}0.94g/dl$ in the Exp. I group, and $6.41{\pm}0.68g/dl$ in the Exp. II group. There was no significance in total protein between the Exp. II group and the Exp. I group from the statistical analysis. 4. The content of albumin in the blood serum was $2.94{\pm}0.13g/dl$ in the normal group, $2.01{\pm}0.48g/dl$ in Chang Tong-young the Exp. I group and $3.15{\pm}0.27g/dl$ in the Exp. II group. This fact showed that the Exp. II group had significant increase in the serum albumin from the statistical analysis compared with the Exp. I group. 5. The content of the globulin in the blood serum was $3.19{\pm}0.48g/dl$ in the normal group, $4.70{\pm}1.26g/dl$ in the Exp. I group and $3.26{\pm}0.57g/dl$ in the Exp. II group. There was no significance in the serum globulin between the Exp. II group and Exp. I group from the statistical analysis. 6. In histological finding, because of severe inflammatory reaction, remarkably irregular tissue and large amount of inflammatory cells were found in the Exp. I group. But the Exp. II group showed small amount of inflammatory cells, the refrained inflammatory state and even recovering state.

• 식물병연구
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• 제9권2호
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• pp.94-98
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• 2003

느타리버섯에 발생하여 갈색부패병을 일으키는 P. tolaasii 및 P.agarici 를 면역화학적 방법으로 신속하게 검출하기 위한 항체를 생산하였고 면역확산법과 ELISA 로 본 항체의 특이성을 조사하였다. $\alpha$-P tolaasii 항체는 P. tolaasii와 반응하여 선명한 침강선을 형성하지 않아 P. tolaasii 에 대하여만 특이적으로 반응하는 항체임을 알 수 있었다. 한편 $\alpha$-P. agarici 항체는 P.agarici뿐만 아니라 다른 세균에 대해서도 침강선을 형성하지 않는 특징을 나타내었다. 비경합 간접 ELISA의 표준곡선으로부터 P.agarici의 검출에 대하여 최고 발색치의 50% 갑을 나타내는 균의 밀도는 $2{\times}10^4$ cfu/ml로 나타났으며, 검출한계농도는 $2{\times}10^3$ cfu/ml 이었다.${\alpha}$-P. agarici 항체는 P. tolaasii의 특이적인 검출에도 효과를 나타냈다. ${\alpha}$-P. tolaasii, ${\alpha}$-P. agarici 항체를 이용하여 확립한 ELISA법은 직접 균체나 버섯 자실체내 P. tolaasii 와 P. agarici의 신속한 검출을 가능하게 하였고, P. tolaasii 와 P. agarici에 오염된 시료를 간단하게 진단하는데 아주 유용하게 사용될 수 있을 것으로 판단되었다.