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This study tested the hypothesis that administration of deracoxib, a cyclooxygenase-2 specific (COX-2) inhibitor, would result in lower gastric lesion scores than administration of buffered aspirin and gastric lesion scores similar to placebo when administered to healthy dogs for 28 days. Twenty-four, healthy, random source dogs were divided into three groups. Group I received buffered aspirin, 23.6 mg/kg PO q 8h, group II received deracoxib, 1.6 mg/kg PO q 24h and placebo twice daily PO q 8h after deracoxib administration, and group III received placebo PO q 8h. Gastroscopy was performed on days -7, 6, 14, and 28 of treatment. Four regions of the stomach (pylorus, incisura, cardia, and body) were evaluated separately and lesions scored on a scale of 1 (mucosal hemorrhage) to 12 (perforating ulcer) by an observer unaware of which treatments the dogs received. Dogs were observed every 8 hours for vomiting, diarrhea and anorexia. Feces were scored from 1-5 (scores <4 were considered diarrhea). Lesion scores for each group, at each location, and total scores, at each time period, were evaluated for the effects of time and treatment using a Kruskal-Wallis test. Total dog days of vomiting and dog days of diarrhea in each group were compared using a Wilcoxon rank sums test. Significance was determined at p<0.05. Significantly higher median total gastric lesion scores were found in the aspirin group compared to the deracoxib or placebo groups on days 6, 14, and 28. There were no significant differences in median total gastric lesion scores between the deracoxib or placebo groups at any time during the study. There was no location effect on gastric lesion scores and there was no significant change in gastric lesion scores over time in any of the groups during treatment. Significantly more dog-days of vomiting occurred in the aspirin group as compared to the deracoxib group. No significant differences were found between groups for dog-days of diarrhea. In this study, the administration of deracoxib to healthy dogs resulted in significantly lower gastric lesion scores compared to dogs receiving aspirin and lesion scores similar to those receiving placebo. / Master of Science

No / Inflammatory changes, characterized by an increase in pro-inflammatory cytokine production and up-regulation of the corresponding signaling pathways, have been described in the brains of aged rats and rats treated with the potent immune modulatory molecule lipopolysaccharide (LPS). These changes have been coupled with a deficit in long-term potentiation (LTP) in hippocampus. The evidence suggests that anti-inflammatory agents, which attenuate the LPS-induced and age-associated increase in hippocampal interleukin-1ß (IL-1ß) concentration, lead to restoration of LTP. Here we report that atorvastatin, a member of the family of agents that act as inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase, exerts powerful anti-inflammatory effects in brain and that these effects are mediated by IL-4 and independent of its cholesterol-lowering actions. Treatment of rats with atorvastatin increased IL-4 concentration in hippocampal tissue prepared from LPS-treated and aged rats and abrogated the age-related and LPS-induced increases in pro-inflammatory cytokines, interferon-¿ (IFN¿) and IL-1ß, and the accompanying deficit in LTP. The effect of atorvastatin on the LPS-induced increases in IFN¿ and IL-1ß was absent in tissue prepared from IL-4¿/¿ mice. The increase in IL-1ß in LPS-treated and aged rats is associated with increased microglial activation, assessed by analysis of major histocompatibility complex II expression, and the evidence suggests that IFN¿ may trigger this activation. We propose that the primary effect of atorvastatin is to increase IL-4, which antagonizes the effects of IFN¿, the associated increase in microglial activation, and the subsequent cascade of events.

Orientador: Wagner Vilegas / Resumo: Apesar dos diversos relatos do uso de Rhipsalis Gaërtn (Cactaceae) por populações tradicionais no tratamento de uma gama de enfermidades, há pouquíssimos estudos fitoquímicos com esse grupo de cactos e por vezes estes são superficiais, apenas indicando a possível presença de algumas classes de produtos naturais. Ademais, estudos avaliando a atividade biológica de Rhipsalis são escassos. Neste viés, este trabalho objetivou investigar a composição química do gênero Rhipsalis e avaliar possíveis atividades ligadas à sua composição, subsidiando desta forma seu uso popular. Esta dissertação foi dividida e organizada da seguinte forma: a primeira parte apresenta uma introdução geral sobre plantas medicinais e sua importância, assim como uma revisão sobre estudos fitoquímicos em Cactaceae. Em seguida, foram redigidas três seções: na primeira, investigamos a composição química de Rhipsalis teres (Vell.) Steud identificando 5 saponinas derivadas do ácido oleanólico, 2 flavonoides C-glicosilados derivados da apigenina e 2 ácidos fenólicos; na segunda, investigamos a composição química do gênero Rhipsalis identificando 28 saponinas cujo core é o ácido oleanólico e 8 flavonoides, também sugerimos fingerprints para cada espécie avaliada com base nas substâncias majoritárias, os resultados quantitativos obtidos por UPLC-MS foram eficientes na identificação das espécies com base na filogenia; na terceira, avaliamos as atividades antioxidante, antifúngica e anti-inflamatória de espécies de R... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Despite the several reports of the use of Rhipsalis Gaërtn (Cactaceae) by traditional populations in the treatment of a range of diseases, there are very few phytochemical studies with this group and sometimes these are superficial, only indicating the possible presence of some classes of natural products. In addition, there are very few studies evaluating the biological activity of Rhipsalis. Therefore, this work aimed to investigate the chemical composition of the genus Rhipsalis and to evaluate possible activities related to its composition, thus subsidizing its popular use. This dissertation was divided and organized as follows: the first part presents a general introduction on medicinal plants and their importance as well as a review on phytochemical studies in Cactaceae. Then, three sections were written: in the first one, we investigated the chemical composition of Rhipsalis teres, identifying 5 saponins derived from oleanolic acid, 2 C-glycosilated flavonoids derived from apigenin and 2 phenolic acids; in the second section, we investigate the chemical composition of the genus Rhipsalis identifying 28 saponins whose core is oleanolic acid and 8 flavonoids, we also suggested fingerprints for each species evaluated on the basis of the majority compounds, the quantitative results obtained by UPLC-MS were efficient in the identification of the species based on the phylogeny; in the third, we evaluated the antioxidant, antifungal and anti-inflammatory activities of Rhipsal... (Complete abstract click electronic access below) / Mestre

研究目的：本論文之研究主要旨在採用兩種非甾體抗炎藥--布洛芬（IBP）及氟比洛芬（FBP）來考察一項新型的納米粒子製備技術--瞬時納米沉澱技術（FNP）。IBP和FBP具有不同理化性質，但其親油性屬大部分藥物所具的典型親油性 （log P = 2-5）。研究證實IBP和FBP有治療阿爾茨海默氏病的潛在藥效，但在血液中廣泛與血漿蛋白結合，導致其腦血管通透性很低。因此，本研究的另一目的為考察FNP製備的納米處方是否能改善此類藥物的大腦遞送。 / 方法：應用FNP，用多入口渦旋混合器（MIVM）將藥物載入聚乙二醇-聚乳酸（PEG-PLA）的納米粒中。通過改變關鍵工藝流程變量考察了變量對納米粒物理性質及穩定性的影響。使用動態光散射儀測定了納米粒粒徑和粒徑分佈，使用zeta 電位分析測定了粒子表面電荷，使用原子力顯微鏡（AFM）確定了納米粒形態，使用x射線光電子能譜（XPS）分析了粒子表面化學成分，使用高效液相色譜測定了的處方載藥量和包封率。使用MDCK和Caco-2細胞株評估了優化後納米處方的細胞通透性，使用健康小鼠進行了優化后纳米處方的体内腦攝取實驗。 / 結果：IBP和FBP納米粒的粒徑均在30-100 nm的範圍內，粒徑分佈均低於或接近0.2。AFM結果顯示，納米粒具有近球狀形態。由多次線性回歸分析各工藝流程變量對IBP納米粒粒徑的影響所得相對重要性的結果為：PLA對PEG之分子量比 > 過飽和比 > 藥物對聚合物比 > 雷諾數。用相同統計方法分析FBP樣品所得結果顯示，PLA對PEG之分子量比亦為影響粒子粒徑的最重要變量。最穩定的IBP納米處方可以在懸浮液狀態下穩定超過1個月，而FBP納米處方為2天。IBP和FBP納米粒的載藥量和包封率均分別超過25%和70%。XPS，AFM和zeta電位測定結果共同表明納米粒中的PEG均偏重位於粒子表面，而相比之下，IBP納米粒中的PEG較FBP納米粒更加偏向於分佈於粒子表面。優化後的IBP納米粒由聚山梨醇酯80包裹後，與IBP溶液相比，顯著增加了IBP的健康小鼠腦攝取量。 / 結論：應用FNP及MIVM製備的聚合物IBP和FNP納米粒，粒徑小，粒徑分佈窄，重現性高，且有較高的載藥量和包封率。納米粒的粒徑主要取決於所採用的共聚物。IBP 納米粒子明顯優越的物理穩定性可歸功於粒子表面較高的PEG濃度。用聚山梨醇酯80包裹納米粒子對於提高IBP的大腦遞送有決定性作用。 / Objectives: The present thesis work was primarily aimed at assessing a relatively novel nanoparticle (NP) production technology termed flash nanoprecipitation (FNP) using two non-steroidal anti-inflammatory drugs, ibuprofen (IBP) and flurbiprofen (FBP), with different physiochemical properties and lipophilicity typical of most drugs (log P = 2-5). Both model drugs were proven to be of potential benefits to the treatment of Alzheimer’s disease, but exhibited poor brain delivery in vivo which could be ascribed to their extensive binding with plasma proteins in the blood. Therefore another aim of the present thesis was to determine whether FNP-produced NP formulations could enhance the delivery of these drugs into the brain. / Methods: Drugs were loaded into NPs of polyethylene glycol (PEG)-polylactic acid (PLA) copolymers of different molecular weights (MWs) by FNP using a four-stream multi-inlet vortex mixer (MIVM). The influence of several key processing variables on the physical properties and stability of the NPs was investigated. The NP preparations were characterized for particle size and size distribution by dynamic light scattering (DLS) sizing analysis; surface charges by zeta potential measurement; particle morphology by atomic force microscopy (AFM); surface composition by x-ray photoelectron spectroscopy (XPS); and drug loading (DL) and encapsulation efficiency (EE) by high performance liquid chromatography. Optimal IBP NP samples were assessed in vitro for cellular permeability using Caco-2 and MDCK cell lines and in vitro for brain uptake in normal mice. / Results: Both IBP and FBP NPs exhibited mean particle size in the range of 30-100nm and polydispersity below or around 0.2. The particles were nearly spherical in shape, as examined by AFM. Multiple linear regression analysis revealed that the relative impact of the processing variables on the particle size of IBP NPs followed the order: PLA-to-PEG MW ratio > supersaturation ratio > drug-to-copolymer ratio > Reynolds number. Similar statistical analysis for FBP NPs also indicated PLA-to-PEG MW ratio being the most significant determinant of particle size. The most stable IBP and FBP NPs in suspension form could last for over 1 month and 2 days respectively. NPs with DLs > 25% and EEs > 70% could be obtained by FNP. XPS in conjunction with AFM and zeta potential analysis revealed that PEG was located more on the surfaces of both IBP and FBP NPs than in the core, but the surface PEG density was higher for the IBP NPs. Coating of optimal IBP NPs with polysorbate 80 significantly improved the brain uptake of IBP in normal mice, compared to IBP solution. / Conclusion: Polymer-stabilized IBP and FBP NPs with particle size below 100 nm and narrow size distribution can be consistently generated by FNP using the MIVM. The copolymer used is the most important determinant of particle size. The superior physical stability of the IBP NPs can be ascribed to their relatively high surface PEG density. High DLs and EEs are achievable with the FNP process. Nanoparticle coating with polysorbate 80 is critical to enhancing the delivery of IBP to the brain in normal mice. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Zhang, Xinran. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 205-245). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese. / Table of Contents --- p.I / Acknowledgements --- p.VI / Abstract --- p.VIII / 摘要 --- p.X / List of Figures --- p.XII / List of Tables --- p.XVII / List of Abbreviations --- p.XIX / Chapter Chapter One --- Introduction / Chapter 1.1 --- Rationale of the study --- p.1 / Chapter 1.2 --- General review of nanoparticulate drug carrier systems --- p.4 / Chapter 1.2.1 --- Background of nanoscience --- p.4 / Chapter 1.2.2 --- Applications of nanoparticulate drug delivery systems --- p.4 / Chapter 1.2.2.1 --- Improved delivery of poorly water soluble drugs --- p.5 / Chapter 1.2.2.2 --- Targeted drug delivery --- p.6 / Chapter 1.2.2.3 --- Drug delivery across the blood brain barrier --- p.8 / Chapter 1.2.2.4 --- Other drug delivery applications --- p.10 / Chapter 1.2.3 --- Types of nanoparticulate drug delivery systems --- p.10 / Chapter 1.2.3.1 --- Nanocrystals --- p.10 / Chapter 1.2.3.2 --- Solid lipid nanoparticles --- p.11 / Chapter 1.2.3.2.1 --- Preparation methods --- p.12 / Chapter 1.2.3.2.2 --- Drug delivery --- p.12 / Chapter 1.2.3.3 --- Polymeric nanoparticles --- p.14 / Chapter 1.2.3.3.1 --- Preparation methods --- p.15 / Chapter 1.2.3.3.2 --- Drug delivery --- p.17 / Chapter 1.2.4 --- Characterization of nanoparticulate drug delivery systems --- p.20 / Chapter 1.2.4.1 --- Particle size and size distribution --- p.21 / Chapter 1.2.4.2 --- Morphology --- p.21 / Chapter 1.2.4.3 --- Zeta potential --- p.23 / Chapter 1.2.4.4 --- Surface chemical composition --- p.23 / Chapter 1.2.4.5 --- Crystallinity --- p.24 / Chapter 1.3 --- Flash Nanoprecipitation technique --- p.25 / Chapter 1.3.1 --- Mechanism and evolution --- p.25 / Chapter 1.3.2 --- Applications --- p.30 / Chapter 1.4 --- Ibuprofen and flurbiprofen --- p.32 / Chapter 1.4.1 --- General characteristics --- p.32 / Chapter 1.4.2 --- Physicochemical properties --- p.33 / Chapter 1.4.3 --- New therapeutic indications --- p.34 / Chapter 1.5 --- Scope of the thesis --- p.36 / Chapter Chapter Two --- Influence of Processing Variables on the Physical Properties and Stability of Ibuprofen and Flurbiprofen Nanosuspensions / Chapter 2.1 --- Introduction --- p.38 / Chapter 2.2 --- Materials and Methods --- p.39 / Chapter 2.2.1 --- Materials --- p.39 / Chapter 2.2.2 --- Solubility of ibuprofen and flurbiprofen in water and acetone mixtures --- p.39 / Chapter 2.2.3 --- Nanoparticle formulation preparation --- p.40 / Chapter 2.2.3.1 --- Determination of the minimum Reynolds number (Re) for homogenous mixing --- p.40 / Chapter 2.2.3.2 --- Effects of processing parameters on particle size and size distribution of ADCP-protected IBP and FBP nanoparticles. --- p.41 / Chapter 2.2.4 --- Particle size and size distribution measurement --- p.42 / Chapter 2.2.5 --- Statistics --- p.42 / Chapter 2.2.6 --- Assessment of nanosuspension stability --- p.42 / Chapter 2.3 --- Results and discussion --- p.43 / Chapter 2.3.1 --- Solubilities of ibuprofen and flurbiprofen in water and acetone mixtures --- p.43 / Chapter 2.3.2 --- Determination of the minimum Re for homogenous mixing --- p.44 / Chapter 2.3.3 --- Effects of processing parameters on particle size and size distribution of the ADCP-protected IBP and FBP nanoparticles. --- p.47 / Chapter 2.3.3.1 --- Effect of solvent type --- p.47 / Chapter 2.3.3.2 --- Effect of PLA-to-PEG MW ratio --- p.64 / Chapter 2.3.3.3 --- Effect of supersaturation --- p.64 / Chapter 2.3.3.4 --- Effect of Re --- p.70 / Chapter 2.3.3.5 --- Effect of drug-to-ADCP ratio --- p.71 / Chapter 2.3.4 --- Effects of processing parameters on the stability of ADCP-stabilized IBP and FBP nanoparticles --- p.72 / Chapter 2.3.4.1 --- Three-day stability --- p.72 / Chapter 2.3.4.2 --- Long-term stability --- p.83 / Chapter 2.4 --- Summary --- p.85 / Chapter Chapter Three --- Drying of Ibuprofen Nanoparticle Suspensions / Chapter 3.1 --- Introduction --- p.86 / Chapter 3.2 --- Materials and Methods --- p.88 / Chapter 3.2.1 --- Materials --- p.88 / Chapter 3.2.2 --- Preparation of IBP nanoparticle formulations with hydrophilic stabilizers or at refrigerated temperature --- p.89 / Chapter 3.2.3 --- Dialysis of nanoparticle formulations --- p.89 / Chapter 3.2.4 --- Freeze-thawing of selected nanoparticle preparations --- p.89 / Chapter 3.2.5 --- Freeze-drying of nanoparticle formulations --- p.90 / Chapter 3.2.6 --- Reconstitution --- p.90 / Chapter 3.2.7 --- Hydrogen bonding coacervate precipitation --- p.91 / Chapter 3.3 --- Results and discussion --- p.91 / Chapter 3.3.1 --- Preparation and dialysis of IBP nanoparticle formulations with hydrophilic stabilizers --- p.92 / Chapter 3.3.2 --- Freeze-drying using cryoprotectants and lyoprotectants --- p.94 / Chapter 3.3.3 --- Freeze-drying with different concentrations of glucose, sucrose and PVA --- p.101 / Chapter 3.3.4 --- Freeze-drying of nanoparticles prepared under other processing conditions --- p.105 / Chapter 3.3.5 --- Hydrogen bonding coacervate precipitation --- p.108 / Chapter 3.4 --- Summary --- p.110 / Chapter Chapter Four --- Physicochemical Characterization of Ibuprofen and Flurbiprofen Nanoparticles / Chapter 4.1 --- Introduction --- p.111 / Chapter 4.2 --- Materials and Methods --- p.112 / Chapter 4.2.1 --- Materials --- p.112 / Chapter 4.2.2 --- Encapsulation efficiency (EE) and drug loading (DL) of IBP nanoparticles --- p.112 / Chapter 4.2.3 --- HPLC analysis of IBP and FBP --- p.113 / Chapter 4.2.4 --- Nanoparticle morphology --- p.114 / Chapter 4.2.4.1 --- SEM --- p.114 / Chapter 4.2.4.2 --- AFM --- p.114 / Chapter 4.2.5 --- Zeta potential measurement --- p.115 / Chapter 4.2.6 --- Surface composition analysis --- p.115 / Chapter 4.3 --- Results and discussion --- p.116 / Chapter 4.3.1 --- Encapsulation efficiency (EE) and drug loading (DL) of IBP nanoparticles --- p.116 / Chapter 4.3.2 --- Nanoparticle morphology --- p.121 / Chapter 4.3.3 --- Surface charges of the nanoparticles --- p.126 / Chapter 4.3.4 --- Surface composition of nanoparticles --- p.128 / Chapter 4.4 --- Summary --- p.145 / Chapter Chapter Five --- Cellular Permeability and In Vivo Brain Uptake of Ibuprofen Nanoparticles / Chapter 5.1 --- Introduction --- p.146 / Chapter 5.2 --- Materials and methods --- p.148 / Chapter 5.2.1 --- Materials --- p.148 / Chapter 5.2.2 --- Methods --- p.148 / Chapter 5.2.2.1 --- Cellular permeability study --- p.148 / Chapter 5.2.2.1.1 --- Cell culture --- p.148 / Chapter 5.2.2.1.2 --- Cell viability study --- p.149 / Chapter 5.2.2.1.3 --- MDCK and Caco-2 cell monolayer permeability assay --- p.150 / Chapter 5.2.2.2 --- In vivo brain uptake study --- p.151 / Chapter 5.2.2.2.1 --- HPLC-UV analysis --- p.151 / Chapter 5.2.2.2.2 --- Preparation of calibration samples --- p.151 / Chapter 5.2.2.2.3 --- Sample preparation --- p.152 / Chapter 5.2.2.2.4 --- Validation of assay methods --- p.153 / Chapter 5.2.2.2.5 --- Animal experiments --- p.154 / Chapter 5.2.2.2.6 --- Data Analysis --- p.155 / Chapter 5.3 --- Results and discussion --- p.155 / Chapter 5.3.1 --- Cellular permeability study --- p.155 / Chapter 5.3.1.1 --- Cell viability study --- p.155 / Chapter 5.3.1.2 --- MDCK and Caco-2 cell monolayer permeability assay --- p.157 / Chapter 5.3.2 --- In vivo brain uptake study --- p.158 / Chapter 5.3.2.1 --- Method validation --- p.158 / Chapter 5.3.2.2 --- Brain uptake of IBP nanoparticles --- p.159 / Chapter 5.4 --- Summary --- p.166 / Chapter Chapter Six --- Conclusions and Future Studies / Chapter 6.1. --- Conclusions --- p.167 / Chapter 6.2. --- Future studies --- p.172 / Appendices --- p.174 / References --- p.205

Schistosome is the causative agent of schistosomiasis which is one of the world's most prevalent tropical diseases. In the skin of infected host, significant inflammatory response to the parasite is not observed. Previous studies from Schistosoma mansoni showed that this subdued inflammatory response was due to a 16-KDa protein, Sm16, which is present abundantly in the secretions of schistosomulae. Provided that Schistosoma japonicum shares the same infective pathway as S. mansoni by penetrating the skin, it seems logical that S. japonicum has a protein with a similar role to Sm16 to down-regulate host immune responses. According to the cDNA sequence of Sm16, a corresponding gene (designated Sj16) of Sm16 has previously been amplified and cloned from the cercarial cDNA of S. japonicum. Sequence analysis showed that Sj16 shares 99% identity with Sm16 in its nucleotide sequence, and 100% identity in its protein sequence. While previous studiers reported their failure in obtaining the soluble recombinant protein of Sm16, we expressed and purified the recombinant Sj16 (rSj16) from E. coli in the present study. Western blot and ELISA analysis showed that S. japonicum-infected rabbit sera could not recognize rSj16, indicating that native Sj16 might fail to induce circulating antibodies during S. japonicum infection. In the in vivo study, rSj16 dramatically suppressed not only the recruitment of leukocytes to the peritoneal cavity of BALB/c mice injected with thioglycollate, but also the maturation of thioglycollate-induced peritoneal macrophages. The suppression effect was accompanied by a marked up-regulation of IL-10 and IL-1RA transcripts, and down-regulation of IL-12p35, IL-1beta and MIP-2 transcripts in peritoneal cells. Further analysis revealed that rSj16 also inhibited both humoral and cellular immune responses to heterologous antigens. In addition, rSj16 was found to induce macrophage differentiation of the murine myeloid leukemia WEHI-3B (JCS) cells, and regulate the differentiation of mouse hematopoietic cells towards the macrophage lineage. Although previous studies indicated the involvement of endogenous IL-1alpha, IL-1beta and TNF-alpha in the macrophage differentiation of JCS cells, the results from this study suggested that rSj16-induced JCS cell differentiation do not rely on the endogenous production of these three cytokines. This is the first study to successfully express and purify sufficient soluble rSj16, and demonstrate the anti-inflammatory and immunomodulatory effects of the rSj16. / Hu, Shaomin. / Adviser: Ming Chiu Fung. / Source: Dissertation Abstracts International, Volume: 71-01, Section: B, page: 0210. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 139-154). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Thesis (M.S.)--University of Michigan, 1993.

Dissertation submitted in partial compliance with the requirements for a Masters Degree in Technology: Chiropractic, Durban University of Technology, 2009. / Headaches are one of the most common clinical conditions in medicine, and 80% of these are tension-type headaches (TTH). TTH has a greater socioeconomic impact than any other type of headache due to its prevalence. Within the TTH category, episodic TTH are more prevalent than chronic TTH. The mainstay in the treatment of TTH are simple analgesics and NSAIDs. Unless contraindicated, NSAIDs are often the most effective treatment for ETTH. However patients suffering with TTH tend to relate their headaches to increased muscle stiffness in the neck and shoulders and thus the non-pharmacological treatment of ETTH could be directed at the associated musculoskeletal components of ETTH. It is therefore proposed that the Kinesio Taping® Method may have an effect in the treatment of the muscular component of ETTH. Method: This study was a prospective randomised clinical trial with two intervention groups (n=16) aimed at determining the relative effectiveness of a NSAID and the Kinesio Taping® Method in the treatment of ETTHs. The patients were treated at 5 consultations over a 3 week period. Feedback was obtained using the: NRS – 101, the CMCC Neck Disability Index and a Headache Diary. Results: The Headache Diary showed a reduction in the presence and number, mean duration and pain intensity of ETTH in both groups. These treatment effects were sustained after the cessation of treatment with the exception of mean pain intensity in the Kinesio Taping® Method group. The mean NRS score decreased in both groups but at a slightly faster rate in the Kinesio Taping® Method group. The CMCC showed an improvement in the functional ability of the patients in both groups. Conclusion: There seems to be no significant difference in the relative effectiveness of the treatment modalities. We can thus state that the overall short-term reduction in symptomatology supports the use of NSAIDs or Kinesio Taping® Method in the treatment of ETTH.

全球過敏性疾病的患病率逐漸增加。大約30 - 40的世界人口患有一個或多個過敏性疾病，它絶對是一個國際性的公共健康問題。在過去三十年，過敏性皮膚炎的發病率增加了2-3 倍，當中患病率最高的是嬰兒和兒童，而且現時並沒有明確的治療方法。然而，對過敏性疾病有效的治療方法仍然缺乏，大多數傳統的治療涉及臨床改善，但不針對促進過敏性炎症發病機制中的主要因素。這些傳統的治療方法都有不良副作用。因此，發展一個更安全和非類固醇的治療方式成為了新的趨勢。 / 從過往的臨床試驗中，患有中度至嚴重過敏性皮膚炎的兒童服用由五種中藥制成的Pentaherbs（PHF）膠囊藥丸後， 顯著地改善他們的生活質數，降低了過敏性皮膚炎指數（SCORAD）及減少使用傳統藥物類固醇的份量，更沒有出現任何不良藥物作用。實驗結果指出PHF 有潛力替代類固醇，成為治療過敏性皮膚炎的取代品。在本研究中，我們使用炎症相關的細胞因子IL-33，激活在有或沒有與皮膚成纖維細胞一起培植下的嗜鹼性細胞系KU812 細胞， 來探討了PHF，牡丹皮（DP，PHF 的五種草藥之一）和沒食子酸（GA，牡丹皮的主要成分之一）的抗炎和抗過敏特性。 / 在過敏性炎症中，嗜鹼性粒細胞是一個重要的效應細胞。我們利用細胞因子IL-33 激活嗜鹼性粒細胞系KU812 細胞，並從研究結果發現出PHF，DP 和GA 能有效及顯著地抑制細胞間粘附分子ICAM-1 的表達，炎症相關趨化因子CCL2，CCL5，CXCL8 和促炎細胞因子IL-6 的釋放。這證實出PHF, DP 和GA有抗炎和抗過敏的特性。在進一步的研究中，我們加入一種常用醫治過敏性炎症藥物的合成類固醇地塞米松, 與PHF, DP 和GA 結合使用。從各種組合的不同濃度地塞米松與PHF，DP 和GA 中，我們發現聯合使用低濃度為0.01 微克/毫升的地塞米松和10 微克/毫升GA 可進一步抑制ICAM-1 在KU812 的表達，趨化因子CCL2 和CCL5 釋放。此外，沒食子酸可顯著抑制細胞內信號分子p38 絲裂原活化蛋白激酶 (MAPK)，IκB-α和JNK 的表達。這表明了黏附分子的表達，趨化因子和細胞因子的釋放的抑製是經由p38 MAPK，IκB-α和JNK 訊息傳遞路徑所調節。實驗證實PHF，DP 和GA 具有抗炎和抗過敏的特性，與過往的臨床試驗結果一致。 / 為了更進一步研究沒食子酸和地塞米松在過敏性炎症的發病機制中扮演的角色， 我們建立了一個體外的模仿患者皮膚皮炎症的模型，共同培養嗜鹼性細系KU812 細胞和皮膚成纖維細胞 。我們發現，單沒食子酸的應用已經可以顯著地抑制在KU812 細胞和成纖維細胞表面粘附分子的表達，並減低釋放過敏性炎症相關的趨化因子CCL2，CCL5，CXCL8 和促炎細胞因子IL-6。此外，地塞米松和沒食子酸的結合使用能增強抑制KU812 細胞面上ICAM-1 的表達，和皮膚成纖維細胞面上ICAM-1 和VCAM-1 的表達，與及趨化因子CCL2 和CXCL8，促炎性細胞因子IL-6 的釋放。 / 上述調查結果表明，天然植物產品PHF，DP 和GA 是具有消炎和抗過敏的作用，抑制嗜鹼性粒細胞趨化遷移至發炎處和隨後釋放的過敏性炎症介質，如炎症相關的趨化因子和促炎細胞因子。結果表明，沒食子酸天然植物產品可能是一個潛在的過敏性皮膚炎的治療劑，而沒食子酸和地塞米松的結合使用，可以有效降低在患者治療皮膚炎症時使用地塞米松的劑量。總結，這項研究結果揭示使用天然植物衍生產品具有更安全,高效力和副作用少的一種新治療方式。 / The worldwide prevalence of allergic diseases has been increasing gradually. Around 30 - 40% of the world population suffers from one or more allergic conditions, it is definitely a national public health issue. The incidence of Atopic Dermatitis (AD) has increased by 2-3 folds in the past 3 decades with no definitive cure, where the case is the highest during the early infancy and childhood. However, effective treatments on allergic diseases are still lacking, with most of the traditional treatment involves clinical improvement but not targeting the primary factors promoting the pathogenesis of allergic inflammation. These traditional treatments have undesirable side effects. Therefore, there has been a rising interest in the development of a safer and nonsteroid immunomodulation formula to cure the disease. / From previous clinical trails, it is revealed that children with moderate-to-severe AD treated with traditional Chinese Medicine, Pentaherbs formula (PHF), have significantly improved their quality of life, lowered the Scoring of Atopic Dermatitis (SCORAD) index and the use of topical steroids without any adverse drug effect, suggesting that PHF can be an alternative potential adjunct therapy for AD. In this present studies, we elucidated the in vitro anti‐inflammatory and anti‐allergic activities of PHF, Cortex Moutan / Danpi (DP, one of the five herbs in PHF) and gallic acid (GA, one of the main ingredients in Danpi) using human basophilic KU812 cells, with or without human dermal fibroblast, upon the activation with alarmin inflammation-related cytokine IL-33. / Human basophilic KU812 cells activated by alarmin cytokine IL-33 were used as basophil cell model for study since basophils are a crucial effector cells in allergic inflammation. Our results showed that PHF, DP and GA exhibited the anti-inflammatory and anti‐allergic activities indicated by significant suppressive effects on the intercellular adhesion molecule (ICAM)‐1 expression, the release of inflammation‐related chemokines CCL2, CCL5, CXCL8 and proinflammatory cytokine IL-6 from IL-33‐activated KU812 basophilic cells. The studies were further investigated with the combined use of synthetic steroid dexamethasone which is a common drug for AD. Among various combinations with different concentrations of dexamethasone with PHF, DP and GA, we demonstrated that the combined use of a concentration as low as 0.01 μg/ml dexamethasone and 10 μg/ml GA could further suppress ICAM‐1 expression, chemokines CCL2 and CCL5 release in IL-33 activated KU812 cells. Furthermore, gallic acid could significantly suppress the intracellular signaling molecules p38 MAPK, IκB-α and JNK in KU812 cells, thereby suggesting the underlying mechanisms for the suppressive effect on adhesion molecules expression, and the chemokines and cytokines release. Both in vivo and in vitro experiments show that PHF, DP and GA exhibit the anti-inflammatory and anti‐allergic activities in concordance to the previous clinical trials using PHF on AD children. / In order to further study the involvement of gallic acid and dexamethasone in the pathogenesis of AD, we then established an in vitro skin inflammatory cell model by co‐culturing human basophilic KU812 cells and human dermal fibroblasts mimicking the skin lesions of the AD patients. We revealed that the application of gallic acid alone could already significantly suppress the adhesion molecules expression on KU812 cell and fibroblasts, and the release of AD-related chemokines CCL2, CCL5, CXCL8 and pro-inflammatory cytokine IL-6 from the co‐culture. In addition, the combined use of dexamethasone and gallic acid showed an enhanced suppressive effect on ICAM-1 on KU812, and ICAM-1 and VCAM-1 on fibroblasts, AD‐releated chemokines CCL2 and CXCL8, and pro-inflammatory cytokine IL-6. / The above findings suggest that PHF, DP and GA are anti-inflammatory and anti-allergic natural plant products by suppressing the transmigration of basophils into the inflamed sites and the subsequent release of allergic inflammation mediators e.g. inflammation-related chemokines and proinflammatory cytokines.The results suggest that natural plant product gallic acid could be a potential therapeutic agent in treating skin inflammation in AD, and the combined use of gallic acid with dexamethasone could lower the dosage of dexamethasone used in AD patients. Together, of the results of this study shed light for a novel therapeutic modality of AD using a safer natural plant derived product with high potency and less side effects to treat AD. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Liu, Yan Ping Kelly. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 107-122). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.III / 摘要 --- p.VI / PUBLICATIONS --- p.IX / ABBREVIATIONS --- p.XI / TABLES OF CONTENTS --- p.XIII / Chapter CHAPTER 1: --- General Introduction / Chapter 1.1 --- Allergy --- p.1 / Chapter 1.1.1 --- Definition of Allergy --- p.1 / Chapter 1.1.2 --- Allergic diseases and their prevalence --- p.2 / Chapter 1.1.3 --- Allergic inflammation and its characteristics --- p.2 / Chapter 1.1.4 --- Treatment of allergy --- p.4 / Chapter 1.1.5 --- Atopic Dermatitis --- p.7 / Chapter 1.2 --- Biology of basophils --- p.8 / Chapter 1.2.1 --- Development of basophils --- p.8 / Chapter 1.2.2 --- Morphology and phenotype --- p.9 / Chapter 1.2.3 --- Mast cells and basophils --- p.11 / Chapter 1.2.4 --- Basophils and allergic inflammation --- p.12 / Chapter 1.2.5 --- Human basophilic KU812 cell line --- p.13 / Chapter 1.3 --- Adhesion molecules in allergic inflammation --- p.14 / Chapter 1.3.1 --- Selectins --- p.14 / Chapter 1.3.2 --- Integrins --- p.16 / Chapter 1.3.3 --- Immunoglobulin gene super family --- p.16 / Chapter 1.4 --- Chemokines in allergic inflammation --- p.18 / Chapter 1.4.1 --- C chemokines --- p.18 / Chapter 1.4.2 --- CC chemokines --- p.18 / Chapter 1.4.3 --- CXC chemokines --- p.19 / Chapter 1.4.4 --- CX3C chemokines --- p.19 / Chapter 1.5 --- Cytokines in allergic inflammation --- p.20 / Chapter 1.5.1 --- Proinflammatory cytokines --- p.20 / Chapter 1.5.2 --- Anti-inflammatory cytokines --- p.23 / Chapter 1.6 --- Signal Transduction in allergic inflammation --- p.25 / Chapter 1.6.1 --- Intracellular signaling mechanisms --- p.25 / Chapter 1.6.2 --- RAS-RAF-MAPK pathway --- p.27 / Chapter 1.6.3 --- JAK/STAT pathway --- p.28 / Chapter 1.6.4 --- PI3K-Akt pathway --- p.28 / Chapter 1.6.5 --- NF-κB pathway --- p.28 / Chapter 1.7 --- Aim of Study --- p.29 / Chapter Chapter 2: --- Materials and Methods / Chapter 2.1 --- Materials --- p.32 / Chapter 2.1.1 --- Cell Culture --- p.32 / Chapter 2.1.2 --- Serum Supplements --- p.33 / Chapter 2.1.3 --- Recombinant human cytokine --- p.33 / Chapter 2.1.4 --- Dexamethasone --- p.33 / Chapter 2.1.5 --- Phosphate-buffered saline --- p.34 / Chapter 2.1.6 --- Dimethyl sulfoxide --- p.34 / Chapter 2.1.7 --- Nucleotide-binding oligomerization domain ligands --- p.34 / Chapter 2.1.8 --- BAY 117082 --- p.34 / Chapter 2.1.9 --- Cell surface and intracellular immunofluorescence staining --- p.35 / Chapter 2.1.10 --- In vitro XTT based toxicology assay kit --- p.38 / Chapter 2.1.11 --- Quantitative analysis of inflammatory mediators release --- p.38 / Chapter 2.1.12 --- Natural Products --- p.39 / Chapter 2.1.13 --- Animal Experiment --- p.40 / Chapter 2.2 --- Methods --- p.41 / Chapter 2.2.1 --- Cell Culture --- p.41 / Chapter 2.2.2 --- Preparation of plant extracts --- p.42 / Chapter 2.2.3 --- Cell toxicity of the natural products --- p.42 / Chapter 2.2.4 --- Flow cytometric analysis of cell surface expression of molecules --- p.43 / Chapter 2.2.5 --- CBA assay --- p.43 / Chapter 2.2.6 --- Flow cytometric analysis of activated intracellular molecules --- p.44 / Chapter 2.2.7 --- Allergic asthmatic mice model --- p.45 / Chapter 2.2.8 --- Statistical analysis --- p.45 / Chapter Chapter 3: --- Anti-inflammatory and anti-allergic properties of Pentaherbs formula, Danpi and Gallic acid / Chapter 3.1 --- Introduction --- p.46 / Chapter 3.1.1 --- Basophils in inflammation --- p.46 / Chapter 3.1.2 --- IL-33 --- p.47 / Chapter 3.1.3 --- Natural plant products --- p.48 / Chapter 3.1.4 --- Dexamethasone --- p.51 / Chapter 3.1.5 --- Hypothesis and aim of study --- p.52 / Chapter 3.2 --- Results --- p.54 / Chapter 3.2.1 --- Cell cytotoxicity of PHF, DP and GA on human basophilic KU812 cells --- p.54 / Chapter 3.2.2 --- Effect of adhesion molecules expression on IL-33-activated KU812 cells treated with PHF, DP and GA --- p.56 / Chapter 3.2.3 --- Effect of PHF, DP and GA on inflammation-related chemokines CCL2,CCL5, CXCL-8 production from IL-33-activated KU812 cells --- p.59 / Chapter 3.2.4 --- Effect of PHF, DP and GA on pro-inflammatory cytokine IL-6 production from IL-33-activated KU812 cells --- p.64 / Chapter 3.2.5 --- Intracellular signaling pathways involved in GA treatment on IL33-activated KU812 cells --- p.67 / Chapter 3.2.6 --- Effect on the adhesion molecules expression, chemokines and cytokines release of IL-33-activated human basophilic KU812 cells upon the combined treatment of PHF/DP/GA with dexamethasone --- p.73 / Chapter 3.2.7 --- In vivo effect of PHF and DP on Th2 and inflammatory cytokines concentration in serum or BALF in allergic inflammatory mice models --- p.76 / Chapter 3.3 --- Discussion --- p.79 / Chapter Chapter 4: --- Gallic acid and Dexamethasone in Atopic Dermatitis / Chapter 4.1 --- Introductions --- p.84 / Chapter 4.1.1 --- Atopic Dermatitis --- p.84 / Chapter 4.1.2 --- Basophils in AD --- p.86 / Chapter 4.1.3 --- Dermal fibroblasts in AD --- p.86 / Chapter 4.1.4 --- Hypothesis --- p.87 / Chapter 4.2 --- Results --- p.88 / Chapter 4.2.1 --- Effect of the combined use of GA and dexamethasone on ICAM-1 expression on KU812 cells co-cultured with fibroblasts --- p.88 / Chapter 4.2.2 --- Effect of the combined use of GA and dexamethasone on ICAM-1 and VCAM-1 expression on fibroblasts co-cultured with KU812 cells --- p.90 / Chapter 4.2.3 --- Effect on chemokines release from the co-culture upon the treatment with GA and dexamethasone --- p.93 / Chapter 4.2.4 --- Effect on cytokine release from the co-culture treated with GA and dexamethasone --- p.96 / Chapter 4.3 --- Discussions --- p.98 / Chapter Chapter 5: --- Concluding Remarks and Future Prospective / Chapter 5.1 --- Concluding remarks --- p.100 / Chapter 5.2 --- Future prospective --- p.101 / References --- p.107