• Journal of Veterinary Science
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• 제21권3호
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• pp.46.1-46.18
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• 2020

Background: High concentrations of particulate matter less than 2.5 ㎛ in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. Objectives: In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. Methods: High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. Results: Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. Conclusions: The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.

• Journal of Veterinary Science
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• 제24권4호
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• pp.52.1-52.13
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• 2023

Background: Mesenchymal stem cells (MSCs) have been investigated as therapeutic agents for inflammatory bowel disease (IBD). Stimulation of MSCs with pro-inflammatory cytokines is an approach to enhance their immunomodulatory effects. However, further investigation is required to support their application in immune-mediated disorders and companion animals. Objectives: This study aimed to assess the therapeutic effect of tumor necrosis factor (TNF)-α-stimulated feline adipose tissue-derived MSCs (fAT-MSCs) in a dextran sulfate sodium (DSS)-induced colitis mouse model. Methods: Colitis mice was made by drinking water with 3% DSS and fAT-MSCs were injected intraperitoneally. Colons were collected on day 10. The severity of the disease was evaluated and compared. Raw 264.7 cells were cultured with the conditioned medium to determine the mechanism, using quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results: TNF-α-stimulated fAT-MSCs more improved severity of DSS-induced colitis in disease activity, colon length, histologic score, and inflammatory cytokine. In sectionized colon tissues, the group comprising TNF-α-stimulated fAT-MSCs had higher proportion of CD11b⁺CD206⁺ macrophages than in the other groups. In vitro, TNF-α-stimulation increased cyclooxygenase-2 (COX-2) expression and prostaglandin E₂ (PGE₂) secretion from fAT-MSCs. The conditioned medium from TNF-α-stimulated fAT-MSCs enhanced the expression of interleukin-10 and arginase-1 in LPS-activated Raw 264.7 cells. Conclusions: These results represent that TNF-α-stimulated fat-mscs ameliorate the inflamed colon more effectively. Furthermore, we demonstrated that the effectiveness was interlinked with the COX-2/PGE₂ pathway.

• IMMUNE NETWORK
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• 제24권2호
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• pp.17.1-17.16
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• 2024

We have reported that anterior cruciate ligament (ACL) injury leads to the differential dysregulation of the complement system in the synovium as compared to meniscus tear (MT) and proposed this as a mechanism for a greater post-injury prevalence of post traumatic osteoarthritis (PTOA). To explore additional roles of complement proteins and regulators, we determined the presence of decay-accelerating factor (DAF), C5b, and membrane attack complexes (MACs, C5b-9) in discarded surgical synovial tissue (DSST) collected during arthroscopic ACL reconstructive surgery, MT-related meniscectomy, osteoarthritis (OA)-related knee replacement surgery and normal controls. Multiplexed immunohistochemistry was used to detect and quantify complement proteins. To explore the involvement of body mass index (BMI), after these 2 injuries, we examined correlations among DAF, C5b, MAC and BMI. Using these approaches, we found that synovial cells after ACL injury expressed a significantly lower level of DAF as compared to MT (p<0.049). In contrast, C5b staining synovial cells were significantly higher after ACL injury (p<0.0009) and in OA DSST (p<0.039) compared to MT. Interestingly, there were significantly positive correlations between DAF & C5b (r=0.75, p<0.018) and DAF & C5b (r=0.64 p<0.022) after ACL injury and MT, respectively. The data support that DAF, which should normally dampen C5b deposition due to its regulatory activities on C3/C5 convertases, does not appear to exhibit that function in inflamed synovia following either ACL injury or MT. Ineffective DAF regulation may be an additional mechanism by which relatively uncontrolled complement activation damages tissue in these injury states.

• IMMUNE NETWORK
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• 제22권2호
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• pp.18.1-18.15
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• 2022

Dysfunction of mitochondrial metabolism is implicated in cellular injury and cell death. While mitochondrial dysfunction is associated with lung injury by lung inflammation, the mechanism by which the impairment of mitochondrial ATP synthesis regulates necroptosis during acute lung injury (ALI) by lung inflammation is unclear. Here, we showed that the impairment of mitochondrial ATP synthesis induces receptor interacting serine/threonine kinase 3 (RIPK3)-dependent necroptosis during lung injury by lung inflammation. We found that the impairment of mitochondrial ATP synthesis by oligomycin, an inhibitor of ATP synthase, resulted in increased lung injury and RIPK3 levels in lung tissues during lung inflammation by LPS in mice. The elevated RIPK3 and RIPK3 phosphorylation levels by oligomycin resulted in high mixed lineage kinase domain-like (MLKL) phosphorylation, the terminal molecule in necroptotic cell death pathway, in lung epithelial cells during lung inflammation. Moreover, the levels of protein in bronchoalveolar lavage fluid (BALF) were increased by the activation of necroptosis via oligomycin during lung inflammation. Furthermore, the levels of ATP5A, a catalytic subunit of the mitochondrial ATP synthase complex for ATP synthesis, were reduced in lung epithelial cells of lung tissues from patients with acute respiratory distress syndrome (ARDS), the most severe form of ALI. The levels of RIPK3, RIPK3 phosphorylation and MLKL phosphorylation were elevated in lung epithelial cells in patients with ARDS. Our results suggest that the impairment of mitochondrial ATP synthesis induces RIPK3-dependent necroptosis in lung epithelial cells during lung injury by lung inflammation.

• Tuberculosis and Respiratory Diseases
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• 제47권1호
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• pp.13-25
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• 1999

연구배경: 결핵은 대식세포와 T 림프구가 주로 관여하는 세포성 면역에 의하여 발생되는 대표적인 질환이다. 특히 Th1 또는 Th2 림프구 가농에 의하여 이루어지는 면역반응의 결과에 따라 결핵균에 대한 감수성 또는 저항성이 결정된다. 본 연구는 치료실패 폐결핵환자의 말초혈액단핵구를 PPD 또는 수용성 TSP 항원으로 결핵의 보호면역과 관계가 있는 Th1 반응과 결핵의 감수성과 관계가 있다고 알려진 Th2 반응을 관찰하였다. 방법: 수용성 TSP 항원과 대조항원인 PPD 항원으로 건강인, 폐결핵으로 진단되어 단기치료지침에 의하여 균음 전화된 치료반응 환자 및 치료실패 환자의 말초혈액 단핵구를 대상으로 단핵구 증식반응과 Th1 반응 및 Th2 반응과 각각 관계가 있는 IFN-$\gamma$ 및 RANTES와 MCP-1 mRNA 발현 빈도를 역전사효소 중합효소연쇄반응으로 조사하였다. 결과: PPD 피부반응 양성 건강인 모두는 PPD 또는 TSP 항원에 의하여 자극지수 4 이상의 유의한 증식반응을 보였으나 PPD 음성 건강인 모두는 PPD 또는 TSP 항원에 의하여 자극지수 4 미만의 증식반응을 보였다. 치유된 환자는 80%의 증식반응을 보였으나 치료실패 환자는 PPD 에 의하여 30.8% 그리고 TSP 항원에 의하여 15.4% 만이 자극지수 4 이상의 유의한 증식반응을 보였다. 치유된 환자의 1FN-$\gamma$ mRNA 발현빈도는 90.0% 이었으나 치료실패 환자는 PPD 또는 TSP 항원에 의하여 23.1% 만이 유도되었다. PPD 양성 건강인의 말초혈액단핵구를 PPD, TSP 또는 PHA로 자극하면 RANTES가 모두 발현되었다. 치료실패 환자, PPD 피부반응 음성 및 치유된 환자를 PPD로 발현을 유도한 경우 각각 76.9%, 80.0%로 대상간에 차이가 없었다. 그러나 TSP 항원으로 유도하면 건강 대조군 및 치유된 환자에 비하여 치료실패 환자는 46.2%로 발현빈도가 유의하게 감소되었다. 또한 PHA로 자극한 경우에서도 치료실패 환자는 69.2%로 감소하여 IFN-$\gamma$ mRNA 발현율 감소 경향과 유사하였다. 치료실패 환자는 MCP-1의 발현빈도가 치유된 환자에 비하여 유의하게 증가되었다. 치료실패 환자에 있어서 PHA 자극의 53.8% 보다는 PPD 또는 TSP로 자극한 경우에 발현빈도가 각각 76.9%로 높았다. 그러나 PPD 양성 건강인 및 치유된 환자는 PPD 또는 TSP로 유도한 결과 40% 이었으며 PHA로 유도한 경우는 각각 80%와 90%로 결핵균 항원에서 낮은 발현 빈도를 보여 치료실패 환자와 상반되는 결과를 보였다. 결론: 치료실패 환자는 PPD 피부반응 양성 건강인 및 치유된 환자에 비하여 말초혈액단핵구의 증식능, 1FN-$\gamma$ 및 RANTES mRNA 발현빈도가 현저히 감소되어 Th1 반응이 억제되어 있었다. 반면에 MCP-1 mRNA의 발현빈도는 현저히 증가되어 Th2 반응의 증가로 결핵균 사균 능력이 치료실패 환자는 감소되어 있다고 생각된다.

• Journal of Nutrition and Health
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• 제41권2호
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• pp.141-146
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• 2008

생체 내 (ex vivo) 실험에서 혼합 (고들빼기, 돌미나리, 메밀, 톳, 생강)시료의 물 추출물을 4주간 격일로 마우스 체중 kg 당 0, 50, 500 mg/kg B.W.의 농도로 마우스에 경구 투여한 후 비장세포 증식능, LPS에 의해 활성화된 복강 대식세포가 분비하는 염증성 사이토카인 ($IL-1{\beta}$, IL-6, $TNF-{\alpha}$)의 생성량을 측정하였다. 그 결과 혼합시료의 물추출물은 ConA나 LPS로 자극하지 않은 경우 50 mg./kg B.W. 농도에서 대조군에 비해 유의적으로 높은 비장 증식능을 보였다. LPS에 의해 활성화된 복강 대식세포의 사이토카인 분비량을 측정한 결과에서도 $IL-1{\beta}$, IL-6, $TNF-{\alpha}$의 분비량은 마우스 체중 kg 당 50 mg/kg B.W. 농도의 투여군에서 높은 생성량을 보여주었으나, 500 mg/kg B.W.의 농도 투여군에서는 IL-6 분비량은 대조군에 비해 높은 반면 $TNF-{\alpha}$의 분비량은 큰 차이를 보이지 않았다. 이상의 결과에 따라 비장세포 증식능과 사이토카인 분비능을 검색한 결과 고들빼기, 도미나리, 메밀, 톳, 생강을 혼합한 혼합시료의 추출물 50 mg/kg B.W. 농도 투여가 면역 세포와 면역 기관의 주요기능을 증진시키리 가능성이 있을 것으로 사료된다. 이러한 연구결과를 토대로 앞으로 혼합식을 통한 기능성 식품 개발에 기초 연구 자료가 될 것으로 기대된다.

• Journal of Animal Science and Technology
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• 제50권2호
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• pp.185-198
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• 2008

급성기 반응 중인 육계 병아리에서 미역 2.0% 사료가 영양소 대사, 항산화 효소활성 및 세포성 면역에 미치는 영향이 조사 되었다. 갓 부화한 숫 육계 병아리(Ross)는 4주간의 실험 사육기간에 미역 0.0%(기초) 및 2.0% 사료를 급여하고, 8, 10, 및 12일령에 Salmonella typhymurium lipopolysaccharide(LPS)를 복강내 주입하여 급성기 반응을 활성화 하였다. 실험사육 전기간을 통한 생산성은 사료 중 미역 함량이나 급성기 반응 또는 그 경험의 영향이 없었다. 대조(정상) 병아리에 비하여, 급성기 반응은 미역 2.0% 사료에서 일당 증체량에 영향이 없었으나, 혈장 과산화물 분해효소, 세포외액 SOD(EcSOD) (p<0.05), 적혈구 MnSOD와 CuZnSOD 활성을 높이고, 그리고 사료 중 미역 함량과 관계없이 대사체중(kg0.75) 당 NB와 ME이용량를 낮추고 TNF-α와 ovotransferrin 활성을 높였다. 미역 0.0% 사료에 비하여, 미역 2.0% 사료는, 급성기 반응과 관계없이 kg0.75 당 NB를 증가시켰고, 급성기 반응 중에는 kg0.75당 UAN 배설에 영향이 없었으나, 혈장 과산화물 농도와 적혈구 세포액의 과산화물 분해효소와 MnSOD(p<0.05) 활성을 낮추고 혈장 과산화물 분해효소와 EcSOD 활성은 높이고(p<0.05), 4주령 병아리에서 LPS 자극 PBMC에서 분비한 IL-1 활성을 증가시켰다.(주 단어:미역, LPS, 단백질과 에너지대사, 항산화계, 친염증성 사이토카인, PBMC, 육계 병아리)

• IMMUNE NETWORK
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• 제2권1호
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• pp.41-48
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• 2002

Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

• 한국식품영양과학회지
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• 제38권11호
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• pp.1535-1542
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• 2009

수삼과 버섯의 새로운 용도를 개발하기 위하여, 수삼추출물이 첨가된 액체배지에서 배양된 노루궁뎅이 균사체 심부발효물의 열수추출물로부터 조다당획분을 조제하였다. 수삼추출물이 액체배지 전체양의 1%, 3%와 5% 첨가된 노루궁뎅이버섯 균사체 심부발효물의 열수추출물로부터 조다당획분(HE-GE-CP-1, 3과 5)이 분획되었을 때, 수율은 각각 5.7, 5.1과 4.8%이었다. 이러한 조다당획분 중 HE-GE- CP-5(saline 대조군의 1.89배)는 HE-GE-CP-1(1.64배)과 HE-GE-CP-3(1.76배)보다 유의적으로 높은 비장세포의 마이토젠 활성을 보여주었다. 또한 HE-GE-CP-5(1.83배)는 소장의 국소 면역조직인 Peyer's patch를 경유한 골수세포 증식활성에서도 HE-CP(1.59배), HE-GE-CP-1(1.44배)과 HE-GE-CP-3(1.69배)보다 높은 활성을 나타내었으며, 항암효과로서의 암전이 억제활성 측정에서는 수삼추출물이 5% 첨가된 조다당획분에서 72.4%의 높은 암전이 억제활성도 보여주었다. 그러나 복강 내 마크로파지의 lysosomal phosphatase의 활성측정에서는 HE-GE-CP-3에서 2.03배의 가장 높은 마크로파지 자극활성을 확인할 수 있었다. 한편, 이러한 면역 및 암전이 억제활성을 갖는 조다당획분인 HE-GE-CP-5는 중성당(63.2%)과 함께 상당량의 산성당(19.3%)을 함유하였으며 단백질은 8.8%가 구성분으로 분석되었다. 이러한 결과로부터 HE-GE-CP-5는 암전이를 억제하기 위한 면역계를 자극할 수 있음이 확인되었으며, 이러한 암전이 억제의 항종양 활성은 마크로파지, 비장세포 및 Peyer's patch cell 등의 활성화 기작과 연계되어 있음을 추정할 수 있었다.

• 한방안이비인후피부과학회지
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• 제15권2호
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• pp.169-182
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• 2002

Background: Allergic rhinitis(AR) is a heterogeneous disorder that despite its high prevalence is often undiagnosed. It is characterized by one or more symptoms including sneezing, itching, nasal congestion, and rhinorrhea. And it is frequently accompanied by symptoms involving the eyes, ears, and throat, including postnasal drainage. There are many different causes of rhinitis in children and adults. Approximately 50$\%$ of all cases of rhinitis are caused by allergy. In the case of rhinitis caused by allergens, symptoms arise as a result of inflammation induced by a gamma globulin E-mediated immune response to specific allergens such as pollens, molds, animal dander, and dust mites. The immune response involves the release of inflammatory mediators and the activation and recruitment of cells to the nasal mucosa. AR is similar to 鼻？, hypersensitive rhinitis in Oriental Medicine. I think hypersensitive rhinitis is including of AR, vasomotor rhinitis and non-allergic rhinitis related with eosinophil increased and so on. Purpose: To perform a clinical analysis of hypersensitive rhinitis including allergic rhinitis and estimate the efficacy of Oriental Medical treatment. Objective: We studied 96 patients who had visited our hospital with complaints of nasal symptoms from March 2000 to February 2002; they had the signs more than 2 - nasal obstruction, watery discharge, sneezing and eye or nasal itching. Parameters Observed & Methods: We treated them with acupuncture & herb-medication. Sometime they used aroma oil or external medicine. 1) the distribution of sex & age groups 2) the clinical type based on duration & the severity of symptom 3) the breakdown of complication & pasl history of Otolaryngologic or allergic disease 4) the clinical assessment and classification of rhinitis(sneezers and runners & blockers) 5) the associated symptoms and signs 6) the classification of Byeonjeung 7) the classification of prescriptions and 8) the efficacy of treatment. Result: 1. In the clinical type of based on duration, the intermittent type was 42.7$\%$ and the persistent was 57.3$\%$. 2. We observed the severity of symptoms based on the quality of life. The mild type was 24.0$\%$ and the moderate-severe was 76.0$\%$. 3. In the clinical assessment and classification of rhinitis, the sneezers and runners type was 69.8$\%$ and the blockers was 30.2$\%$. 4. The most common family history with otolaryngologic or allergic disease were allergic rhinitis(17.7$\%$), urticaria, paranasal sinusitis and T.B.(3.1$\%$). 5. The most common past history with otolaryngologic or allergic disease were paranasal sinusitis(14.6$\%$), atopic dermatitis and asthma(8.3$\%$). It was 31.3$\%$ they had a family history and 44.8$\%$, past history. 6. The most common complication was paranasal sinusitis(15.6$\%$). In decreasing order the others were otitis media with effusion(9.4$\%$), GERD and headache(6.3$\%$), asthma, bronchitis, nasal bleeding and allergic dermatitis(5.2$\%$). 7. Classification through Byeonjeung : ⅰ) 39 cases(34.9$\%$) were classified as showing Deficiency syndrome. The insuffficiency of Qi was 17.7$\%$, deficiency of Kidney-Yang, 12.5$\%$ and Lung-Cold, 10.4$\%$. ⅱ) 57 cases(59.4$\%$) were classified as showing Excess syndrome. The Fever of YangMing-meridian was 35.4$\%$, Lung-Fever, 24.0$\%$. 8. The efficacy of treatments showed: an improvement in 22cases(22.9$\%$); an improvement partly in 24 cases(25.0$\%$); no real improvement or changes in 16 cases(16.7$\%$); and couldn't check the results 18cases(18.6$\%$). Conclusion: We suggest that this study could be utilized as a standard of clinical Oriental Medical treatment when we treat hypersensitive rhinitis including allergic rhinitis.