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針灸抗腫瘤免疫臨床試驗文獻研究楊靜一, 11 June 2016 (has links)
2012年,全球大約有1410萬人患上癌症, 820萬人死於癌症。我國惡性腫瘤發病率亦高達235.23/10萬。目前公認的放化療不僅存在副作用,且嚴重影響患者免疫功能,與之不同的是,針灸干預並非直接作用於腫瘤,而是機體的神經-內分泌-免疫網路、心理(安慰)甚至基因方面,使機體產生抗腫瘤的效應,其中提高機體的免疫力即相當於中醫的“扶正固本。本研究擬以針灸抗腫瘤免疫臨床研究試驗中,分析放化療基礎上運用針灸療法的臨床療效,以期進一步臨床指導。 方法:選用國內權威數據庫中國期刊全文數據庫( CNKI )、中國優秀碩士學位論文全文數據庫、中國博士學位論文全文數據庫、維普中文期刊數據庫( VIP )、萬方學術期刊全文數據庫、中國生物醫學文獻服務系統( CBM )以及MEDLINE(OVID)等作為資料來源。選擇在針對放他療治療基礎上運用針灸療法(治療組)抗腫瘤免疫的臨床試驗文獻,進行穴位頻次等基礎數據的綜合分析,並將經Jadad量表評分2分以上的文獻納入系統評價(Meta分析) 結果:經嚴格篩選,最終納入文獻64篇,涉及4286例患者,進行基礎數據的綜合分析並從中選擇品質較高的18篇文獻進行系統評價。從統計結果分析,與對照主比較,治療組在抗腫瘤提升免疫功能方面具有統計學意義, 證實針灸抗腫瘤免疫的療效肯定。 結論:針灸抗腫瘤免疫治療優勢明顯,對各型腫瘤存在廣泛治療作用,但能否實現對某一型腫瘤或某一類免疫細胞靶向性治療仍有待研究。故還需要設計良好的隨機對照及多中心臨床試驗做進一步探討,為尋找腫瘤治療的新出路提供依據。
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Developing new immuno-oncology drugs from traditional Chinese medicineLi, Yang 28 October 2020 (has links)
The most exciting area in current cancer research is immuno-oncology, which aims to develop immunotherapy that activates the human immune system to attack cancers. However, we still lack broadly effective drugs and drug targets for this promising new cancer treatment modality. In an attempt to seek new immuno-oncology drugs that particularly target the antitumor innate immunity, our lab had previously screened traditional Chinese herbal medicine and found that water extract from a medicinal plant, Alocasia Cucullata (AC), has strong anticancer activity in mouse solid tumor models and acts partly by promoting antitumor, proinflammatory macrophages. However, the active components responsible for this exciting immuno-oncology activity and the corresponding immune targets are unknown. Therefore, the aim of my PhD study is to develop chemical biology strategies to isolate and purify the active components of AC from the crude water extract and identify the corresponding cellular targets and mechanisms. Results from my study identified two separable activities and active components, one smaller than 3K and the other larger than 100K, which work synergistically to simulate antitumor macrophages. Further analysis revealed the >100K active component is a large polysaccharide that binds to multiple Toll-like Receptors (TLRs) critical for activating proinflammatory M1-type macrophages. Identity of the Nonetheless, I was able to clean up this fraction by 50 fold and perform RNAseq to examine the innate immune targets of this intriguing drug lead and found it acts to differentiate monocytes to macrophages. Overall my PhD thesis has explored new chemical biology strategies to purify and characterize active components from traditional Chinese medicine towards new drug development and developed a variety of cell-based immune activity assays for identifying and characterizing novel innate immune drug targets and mechanisms
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Complementary and alternative medicine use and perceptions of control among women diagnosed with breast cancerHenderson, Jessica W. 26 June 2001 (has links)
The number of women living with a breast cancer diagnosis will continue to
increase with growing breast cancer incidence rates, greater utilization of early
detection, and longer length of survival times. The prevalence of complementary
and alternative medicine (CAM) is likely to increase as well, making it important to
determine the nature and extent of CAM use in this population. This study
explored CAM use and the influence of the control constructs in the context of the
theory of cognitive adaptation. Computer-assisted telephone interviews were
completed with 551 women diagnosed with breast cancer in Portland, Oregon.
Results indicated that two-thirds (66%) of the women used at least one CAM
therapy during the past 12 months. The majority of women had high perceptions of
cancer control and believed the CAM therapies were important in influencing the
course of the cancer. Logistical regression analysis found that significant
demographic predictors of CAM use were younger age, higher education, and
private insurance. Confirmatory factor analysis was used to refine and test the
construct validation of the Cancer Locus of Control scale. Results supported a
three-factor model (control over cause of cancer, control over course of cancer, and
religious control of cancer) of the scale. Results of multinomial logistical
regression indicated that higher perceptions of control over the course of the cancer
significantly predicted CAM use in three categories. Religious control over the
cancer was not a predictor of CAM use. The findings from this study will help
health care professionals and policy makers identify patient needs that go beyond
surgery, chemotherapy and radiation, and address patient-centered health-related
goals and outcomes for optimal health and recovery from breast cancer. / Graduation date: 2002
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Cytotoxicity and gene expression of selected apoptotic markers in the human laryngeal carcinoma cell line (HEp-2) by Bulbine spp. fractionsSingh, Rishan 30 July 2013 (has links)
Dissertation submitted in fulfilment of the requirements for the Degree of Master of Technology: Biotechnology, Durban University of Technology, 2012. / Apoptosis, or programmed cell death, is a process which is pivotal in eliminating damaged,
infected, or unwanted cells from the body. It has been studied in numerous types of cell lines
ranging from normal to infected cell lines, and there have been a wide range of studies on
apoptosis in laryngeal cancer because this type of cancer has become one of the most
common types of head or neck cancer due to the high incidence of alcohol consumption,
tobacco smoking and chewing of betel quid amongst populations. Laryngeal cancer is usually
treated with radiotherapy or is surgically removed, but due to the loss of the function of the
larynx after surgery, it has been suggested that alternative strategies or ways of treating
laryngeal cancer are required. This has prompted the use of, and research in the field of, plant
medicine to combat laryngeal cancer.
Plant medicine has been used for centuries by the Chinese, Indian and Arabian population in
Uhani, Ayurveda and Siddha as a form of replacing conventional medicine with
complementary and alternative medicine, these include many plants from the family
Asphodelaceae, which have become marketable commodities owing to their medicinal values
and traditional uses. Amongst these plants, the genus Bulbine has been used as a form of
natural medicine in rural Africa and they are also exploited for their aloe vera properties as
well as their possession of phytochemical compounds such as isoflavanoids, nor-lignans,
naphthalene derivatives, anthracene and poly prenylated flavonoids. There has been a
compelling amount of literature on the traditional uses of the Bulbine spp. because these are
linked to the Bulbine spp. having secondary metabolites such as pyroles, chromones,
coumarins, bianthraceane, benzene as well as alkaloids. However, for Bulbine natalensis and
B. frutescens, the plants of interest in this study, the location of anticancer compounds in
them are the only amounts of information available. It has been reported, traditionally, that B.
natalensis possesses the anticancer potential in the roots, while the anticancer potential for B.
frutescens is in the leaves. However, this requires scientific clarification. Therefore, this study
was conducted to assess programmed cell death or apoptosis by analysing the responses of
the human laryngeal carcinoma cell line (HEp-2) to crude aqueous and organic (50% and
100% ethanol) fractions of B. natalensis and B. frutescens. In order to have achieved this, the
HEp-2 cell line was exposed to the above mentioned fractions at three different final
concentrations (20, 2 and 1μg/ml) and assessed for cytotoxicity using the 3-(4,5-
dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay as an
indicator of cell death after fraction utilisation (3 days) for 5 and 8 days. The differences in
the potency of the Bubline spp. fractions were confirmed using the non-parametric ANOVA
test. Thereafter, selected fractions were screened for apoptotic potential using reverse
transcriptase-polymerase chain reaction (RT-PCR) to determine the expression of bax and
caspase-3 biomarkers, which are the biomarkers that participate in mitochondrial,
endoplasmic reticulum and receptor-ligand mechanism of apoptosis. The fractions of B.
frutescens were selected relative to those of B. natalensis for the RT-PCR procedure (read
section 3.4.1. for the selection procedure) and links between the cytotoxicity and gene
expression results were analysed.
It was found that the B. natalensis fractions had a much greater cytotoxic effect on the HEp-2
cell line compared to fractions of B. frutescens by the fifth day of the MTT assay. On the
eight day of incubation, there was an increase in HEp-2 cell line proliferation by the fractions
of both plant species administered. The fractions selected for bax and caspase-3 gene
expression analysis for B. natalensis were the: 20 μg/ml root and corm aqueous fractions, 20
μg/ml leaf and corm 100% ethanol fractions, 20 μ g/ml corm 50% ethanol fraction, 2 μg/ml
root aqueous fraction, 2 μg/ml leaf 100% ethanol fraction and the corm 1 μg/ml aqueous and
50% ethanol fractions. The fractions that were compared to B. natalensis were the 20 μg/ml
root and leaf aqueous and 100% ethanol fractions respectively, the 2 μg/ml root aqueous
fraction and the 2 μg/ml leaf 100% ethanol fraction. It was found from RT-PCR analysis that
all of the B. natalensis fractions tested induced expression of caspase-3, which indicated that
those fractions were capable of inducing apoptosis in laryngeal carcinoma in vitro, since
caspase-3 is the molecular indicator of apoptosis. The aqueous B. frutescens root fraction, did
not induce expression of caspase-3 gene, although it caused expression of bax. This implied
that the root aqueous B. frutescens fraction, may be involved in some other form of cell
death, other than apoptosis. It was also found that there was variability in the response of the
HEp-2 cell line to the Bulbine spp. fractions because of the variation in bax expression among
fractions of different concentration. It was difficult, from this study, to classify fractions into
categories for their mechanism of action, because not all of the fractions that caused the
expression of capase-3, induced bax gene expression. Hence, proper conclusions were unable
to be made, more so, because all the mechanisms of apoptosis mentioned, involve bax gene
activation in order to proceed to completion. Therefore for those Bulbine spp. fractions to
which the HEp-2 cell line exhibited a variable response to, it was postulated that cell death or
apoptosis occurred through some other unknown mechanism. Overall, the cytotoxicity result
didn’t correlate to the gene expression results because fractions that promoted HEp-2 cell line
growth by day five, expressed apoptotic markers, which highlighted the sensitivity and
accuracy of the cells-to-cDNATM II kit for detecting a few possibly apoptosed cells. This was
confirmed by the fact that the HEp-2 cell line used in the MTT cytotoxicity assay and gene
expression study had the same passage number and were viable, the latter being achieved
because the MTT assay only measures the cytotoxicity of compounds once they have been
taken up by viable cells – measuring mitochondrial activities expressed as absorbances.
Therefore, the deduction that HEp-2 cell death may be due to bax/caspase-3 expression was
valid because the mRNA was isolated from viable HEp-2 cells that had been killed by
Bulbine spp. fractions of different polarity. Furthermore, the lack of correlation between the
cytotoxicity and gene expression results indicated the amount of HEp-2 cell line proliferation
by the fraction out-competes those that died, thereby producing a negative cytotoxicity result.
There was a relationship between the traditional information about the anticancer potential
for B. natalensis and B. frutescens. For example, the aqueous root fractions of B. natalensis
were found to be non-toxic to the HEp-2 cell line, but did express caspase-3, which indicated
the possibility of apoptosis. Similarly, the 100% ethanol leaf B. frutescens fractions were
non-toxic to the HEp-2 cell line, but were able to induce apoptosis as well. This emphasised
that the MTT cytotoxicity assay should be compared with other methods of measuring
cytotoxicity when performing studies like this, because although literature has emphasised
many advantages of using the MTT cytotoxicity assay in apoptotic studies, this study proved
otherwise.
When identical HEp-2 cells were treated with the same extract, only some cells were killed
(apoptosis) whereas others proliferated. This was because although the cells were identical
phenotypically, they were all probably at different phases of the cell cycle resulting in the
HEp-2 cells responding variably to the same fraction at different concentrations. It was also
found that the responses were concentration independent. For example, the 1 μg/ml B.
natalensis corm fraction exhibited the highest toxicity of the three concentrations
administered. The lowest cytotoxicity was achieved for the 20 μg/ml fraction – showing a
proliferative effect on the HEp-2 cell line. Similarly, the 2 μg/ml aqueous B. natalensis leaf
fraction induced the highest cytotoxicity level in the HEp-2 cell line followed by the 1 μg/ml
and then the 20 μg/ml fractions. Apart from the genetic variation in identical HEp-2 cells;
this indicated that the HEp-2 cell line was selective to particular fractions of the Bulbine spp.
for utilisation. Concentration independence and HEp-2 cell preferential selection has been
reported in many other studies involving plant fractions/extracts and natural products.
This study demonstrated that although all the tested B. natalensis fractions were capable of
inducing HEp-2 cell death possibly via. apoptosis (caspase-3 induction), a lack of any link
between apoptosis and the cytotoxicity results (hence the 20 μg/ml corm fraction had a
negative cytotoxicity but expressed both apoptotic markers), indicated the need for
phytochemical screening of both Bulbine spp. in future, to determine the compounds that are
responsible for the cytotoxicity and gene expression result outcomes of both Bulbine spp.
fractions. Furthermore, procaspase genes also have to be analysed since genes are expressed
to form procaspases, which then form active caspases.
Although normal cells also express caspase-3 genes during apoptosis, this study focused
exclusively on the effect of Bulbine natalensis and B. frutescens fractions (selected relative to
the cytotoxicity results of B. natalensis) on the HEp-2 cell line (read cell culture and
cytotoxicity discussion for selection of HEp-2 cell line). The validity of this study is
confirmed by similar experimental designs that assayed the cytotoxicity of plant-derived or
natural compounds on cancer cell lines only, and the detection of apoptosis through caspase-
3 induction and other unrelated methods. This is the first study to report the induction of
apoptosis in cancer cell lines by Bulbine spp. fractions using cytotoxicity and the expression
of bax and caspase-3 apoptotic markers. It provides insight into the interaction between the
HEp-2 cell line and the aqueous and organic fractions of B. natalensis and B. frutescens by
analyzing links between cytotoxicity and bax and caspase-3 gene expression; which could
probably contribute to drug design with selected Bulbine spp. fractions. Further investigations
are required in future, to confirm the possible drug targets of the studied Bulbine spp.
fractions in an attempt of assaying their therapeutic importance. / National Research Foundation
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A longitudinal study of emotional distress and the use of complementary and alternative medicine in women with breast cancerShumay, Dianne M January 2005 (has links)
Thesis (Ph. D.)--University of Hawaii at Manoa, 2005. / Includes bibliographical references (leaves 68-75). / Also available by subscription via World Wide Web / ix, 75 leaves, bound 29 cm
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The relationship of belief in control and commitment to life to cancer patients' inclination to use unproven cancer therapiesSkinn, Barbara Jean January 1990 (has links)
The purpose of this study was to explore the relationship of belief in control and commitment to life to the adult cancer patient's inclination to use unproven cancer therapies. A convenience sample of 40 lung cancer patients completed the Wallston's Multidimensional Health Locus of Control Scale, Crumbaugh's Purpose in Life Scale, Hiratzka's Alternative Therapy Scale, and a patient information sheet. The majority of participants exhibited a strong internal locus of control orientation and a strong commitment to life. Belief in control, commitment to life, and the degree of inclination to use unproven cancer therapies were not significantly associated. However, age was negatively correlated with inclination to use unproven cancer therapies. The majority of participants had heard of five or more unproven cancer remedies, and exhibited a strong inclination to use these unorthodox therapies. The most frequently used unproven therapies were anti-medicines - imagery, faith-healing, megadose vitamins, and taheebo. The rising popularity of these anti-medicines has been reported in the literature. The findings were discussed in relation
to theoretical expectations, other research studies, and the methodological problems inherent in the study. Implications of the findings for nursing practice, theory, and education were suggested. Recommendations for further nursing research were made. / Applied Science, Faculty of / Nursing, School of / Graduate
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Anti-cancer effects of the products of Ganoderma lucidum, G. tsugae and their artificial hybrid on breast cancer cells.January 2005 (has links)
Luk Wing Yan Vivien. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 207-239). / Abstracts in English and Chinese. / Acknowledgment --- p.i / Abstract --- p.iii / 摘要 --- p.vi / Contents --- p.viii / List of Figures --- p.xiv / List of Table --- p.xxv / Abbreviations --- p.xxv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Ganoderma spp --- p.1 / Chapter 1.2 --- Bioactive components of Ganoderma spp --- p.3 / Chapter 1.2.1 --- Lingzhi polysaccharide --- p.3 / Chapter 1.2.2 --- Terpenes --- p.4 / Chapter 1.3 --- Ganoderma spp. as Chinese traditional medicine --- p.5 / Chapter 1.4 --- Artificial hybridisation of Ganoderma luciudm and G. tsugae --- p.6 / Chapter 1.4.1 --- Protoplast isolation and fusion of Ganoderma tsugae and G. lucidum --- p.8 / Chapter 1.5 --- Breast Cancer --- p.8 / Chapter 1.5.1 --- Anti-tumor effects of natural substances against breast cancer cell MCF-7 --- p.9 / Chapter 1.5.2 --- Anti-tumor effects of natural substances against breast cancer cell MDA-MB-231 --- p.11 / Chapter 1.5.3 --- Anti-proliferation of cancer --- p.12 / Chapter 1.5.3.1 --- Cell cycle arrest --- p.12 / Chapter 1.5.3.2 --- Cell death --- p.13 / Chapter 1.5.4 --- Anti-proliferation assays --- p.17 / Chapter 1.5.4.1 --- MTT assay --- p.17 / Chapter 1.5.4.2 --- Trypan blue cell viability assay --- p.18 / Chapter 1.5.4.3 --- BrdU assay --- p.18 / Chapter 1.6 --- Endocrine system and hormones --- p.19 / Chapter 1.6.1 --- Estrogen --- p.23 / Chapter 1.6.2 --- Estrogen receptors --- p.24 / Chapter 1.6.3 --- Estrogen action --- p.29 / Chapter 1.6.4 --- Estrogenicity assays --- p.32 / Chapter 1.6.4.1 --- Recombinant yeast assay --- p.33 / Chapter 1.6.4.2 --- E-screen assay --- p.35 / Chapter 1.6.4.3 --- Estrogen receptor competitor binding assay --- p.36 / Chapter 1.6.4.4 --- Endogenous estrogen-regulated gene expression assay --- p.39 / Chapter 1.6.4.4.1 --- Transforming growth factor --- p.39 / Chapter 1.6.4.4.2 --- Monoamine oxidase A --- p.40 / Chapter 1.6.4.4.3 --- pS2 --- p.40 / Chapter 1.6.4.5 --- Uterotrophic assay --- p.41 / Chapter 1.6.4.6 --- Comparison of in vitro and in vivo assay --- p.42 / Chapter 1.7 --- Aim of study --- p.45 / Chapter 1.7.1 --- Objectives --- p.45 / Chapter Chapter 2 --- Materials and Methods --- p.47 / Chapter 2.1 --- Fungal culture --- p.47 / Chapter 2.2 --- Artificial hybridization of Ganoderma tsugae and G. lucidum --- p.47 / Chapter 2.2.1 --- Protoplast isolation of Ganoderma tsugae and G. lucidum --- p.47 / Chapter 2.2.2 --- Protoplast fusion of Ganoderma tsugae and G. lucidum --- p.48 / Chapter 2.3 --- Screening and selection of hybrid ´Ø --- p.49 / Chapter 2.3.1 --- Temperature screening --- p.49 / Chapter 2.3.2 --- DNA fingerprint by Arbitarily-primed polymerase chain reaction --- p.49 / Chapter 2.3.2.1 --- Extraction of genomic DNA --- p.49 / Chapter 2.3.2.2 --- Arbitrarily-primed polymerase chain reaction --- p.50 / Chapter 2.3.2.3 --- Gel electrophoresis --- p.51 / Chapter 2.4 --- Confirmation test --- p.51 / Chapter 2.4.1 --- Somatic incompatibility test --- p.51 / Chapter 2.4.2 --- DNA fingerprinting by specific polymerase chain reaction --- p.52 / Chapter 2.4.2.1 --- Specific Polymerase Chain Reaction (PCR) --- p.52 / Chapter 2.4.2.2 --- Purification of PCR products --- p.52 / Chapter 2.4.2.3 --- Cycle-sequencing --- p.53 / Chapter 2.3.2.4 --- Sequencing --- p.54 / Chapter 2.3.2.5 --- Sequence analysis --- p.54 / Chapter 2.5 --- Characterization of the selected hybrid --- p.56 / Chapter 2.5.1 --- Scanning electron microscopy (SEM) --- p.56 / Chapter 2.5.1.1 --- Preparation of specimens for scanning electron microscopy --- p.56 / Chapter 2.5.1.2 --- "Cytological studies of pileus, stipe and spores of G. lucidum, G. tsugae and hybrid" --- p.57 / Chapter 2.5.2 --- Temperature effect --- p.57 / Chapter 2.5.3 --- Submerged fermentation --- p.57 / Chapter 2.5.4 --- Fruiting test --- p.58 / Chapter 2.6 --- "Bioactive components of G. lucidum, G. tsugae and hybrid" --- p.58 / Chapter 2.6.1 --- Sample preparation --- p.58 / Chapter 2 6.2 --- Lingzhi polysaccharide --- p.59 / Chapter 2.6.3 --- Terpenes --- p.59 / Chapter 2.7 --- Effect of extracts against breast cancer cell lines --- p.60 / Chapter 2.7.1 --- Cell culture --- p.60 / Chapter 2.7.2 --- Lingzhi Extract preparation --- p.61 / Chapter 2.7.3 --- Optimization of cell density --- p.61 / Chapter 2.7.3.1 --- MTT assay --- p.61 / Chapter 2 7.3.2 --- Trypan blue cell viability assay --- p.62 / Chapter 2.7.3.3 --- BrdU assay --- p.62 / Chapter 2.7.3.4 --- Growth curve of MCF-7 --- p.63 / Chapter 2.7.3.5 --- Growth curve of MDA-MB-231 --- p.64 / Chapter 2.7.4 --- Anti-proliferative effect of extracts on MCF-7 cells --- p.69 / Chapter 2.7.4.1 --- MTT assay --- p.69 / Chapter 2 7.4.2 --- Trypan blue cell viability assay --- p.69 / Chapter 2.7.4.3 --- BrdU assay --- p.70 / Chapter 2.7.5 --- Study of cultured medium effect of biomass and pileus extracts on MCF-7 cells --- p.71 / Chapter 2.7.5.1 --- Cultured medium effect ofbiomass and pileus extracts --- p.71 / Chapter 2.7.6 --- mRNA expression assay (RT-PCR) --- p.71 / Chapter 2.7.6.1 --- Effect of extract on gene expression --- p.71 / Chapter 2.7.6.2 --- Time effect of extract on gene expression --- p.72 / Chapter 2.7.6.3 --- Isolation of RNA --- p.72 / Chapter 2.7.6.4 --- Quantification and qualification of DNA and RNA by spectrophotometry --- p.73 / Chapter 2.7.6.5 --- First strand cDNA synthesis --- p.73 / Chapter 2.7.6.6 --- Amplification of cDNA --- p.74 / Chapter 2.7.7 --- Effect of biomass and pileus lingzhi polysacchandes and terpenes on MCF-7 cells --- p.75 / Chapter 2.7.7.1 --- Effect of reconstitution of lingzhi polysacchande and terpenes on MCF-7 cells --- p.75 / Chapter 2.7.8 --- Effect of biomass and pileus extracts on MDA-MB-231 cells --- p.76 / Chapter 2.8 --- Estrogenicigy assay --- p.76 / Chapter 2 8.1 --- E-screen test --- p.76 / Chapter 2.8.2 --- Estrogen receptor competitor binding assay --- p.77 / Chapter 2.8.3 --- pS2 mRNA expression assay --- p.78 / Chapter 2.9 --- DNA microarray analysis --- p.79 / Chapter 2.9.1 --- mRNA purification --- p.79 / Chapter 2.9.2 --- RT and LPR (Linear Polymerase Reaction) labeling --- p.80 / Chapter 2 9.3 --- pre-hybridization --- p.81 / Chapter 2.9.4 --- Hybridization --- p.82 / Chapter 2.9.5 --- Detection --- p.82 / Chapter 2.9.6 --- Image acquisition and analysis --- p.83 / Chapter Chapter 3 --- Result --- p.84 / Chapter 3.1 --- Artificial hybndization of Ganoderma tsugae and G. lucidum --- p.84 / Chapter 3.1.1 --- protoplast isomation and fusion of Ganoderma tsugae and G. lucidum --- p.84 / Chapter 3.2 --- Screening and selection of hybrid --- p.84 / Chapter 3.2.1 --- Temperature screening --- p.84 / Chapter 3.2.2 --- DNA fingerprint by Arbitrarily-primed polymerase chain reaction --- p.86 / Chapter 3.3 --- Confirmation tests --- p.88 / Chapter 3.3.1 --- Somatic incompatibility test --- p.88 / Chapter 3.3.2 --- DNA fingerprinting by specific polymerase chain reaction --- p.90 / Chapter 3.4 --- Characterization of selected hybrid --- p.100 / Chapter 3.4.1 --- Scanning electron micscropy --- p.100 / Chapter 3.4.2 --- Temperature effect --- p.103 / Chapter 3.4.3 --- Submerged fermentation --- p.105 / Chapter 3.4.4 --- Fruiting test --- p.107 / Chapter 3.5 --- "Bioactive components of G. lucidum, G. tsugae and hybrid" --- p.109 / Chapter 3.5.1 --- Lingzhi polysaccharide --- p.109 / Chapter 3.5.2 --- Terpenes --- p.109 / Chapter 3.6 --- Effect of extracts against breast cancer cell lines --- p.119 / Chapter 3.6.1 --- Anti-proliferative effect of extracts on MCF-7 cells --- p.119 / Chapter 3.6.2 --- Study of medium effect of biomass and pileus extracts on MCF-7 cells --- p.139 / Chapter 3.6.3 --- mRNA expression assay (RT-PCR) --- p.143 / Chapter 3.6.4 --- Effect of biomass and pileus lingzhi polysaccharides and terpenes on MCF-7 cells --- p.150 / Chapter 3.6.5 --- Effect of biomass and pileus extracts on MDA-MB231- cells --- p.159 / Chapter 3.7 --- Estrogenicity assay --- p.166 / Chapter 3.7.1 --- E-screen assay on biomass and pileus extracts --- p.166 / Chapter 3.7.2 --- E-screen assay on biomass and pileus terpenes and lingzhi polysaccharide --- p.166 / Chapter 3.7.3 --- Estrogen receptor competitor binding assay --- p.169 / Chapter 3.7.4 --- pS2 mRNA expression assay --- p.175 / Chapter 3.8 --- DNA microarray analysis --- p.177 / Chapter Chapter 4 --- Discussion --- p.184 / Chapter 4.1 --- Artificial hybridization of Ganoderma tsugae and G. lucidum --- p.184 / Chapter 4.1.1 --- Protoplast isolation and fusion of Ganoderma tsugae and G. luciudm --- p.184 / Chapter 4.1.2 --- Screening and selection of hybrid --- p.184 / Chapter 4.1.3 --- Characterization of the selected hybrid --- p.185 / Chapter 4.1.4 --- "Nature of hybrid, mutant and variant" --- p.189 / Chapter 4.2 --- Effect of extracts against breast cancer cell lines --- p.190 / Chapter 4.2.1 --- Anti-proliferative effect of extracts on MCF-7 cells --- p.190 / Chapter 4.2.2 --- Study of effect of cultured medium of biomass and pileus extracts on MCF-7 cells --- p.193 / Chapter 4.2.3 --- Effect of biomass and pileus extracts on MDA-MB231- cells --- p.194 / Chapter 4.2.4 --- mRNA expression assay (RT-PCR) --- p.195 / Chapter 4.3 --- Estrogenicity --- p.198 / Chapter 4.3.1 --- E-screen assay --- p.198 / Chapter 4.3.2 --- Estrogen receptor competitor binding assay --- p.199 / Chapter 4.3.3 --- pS2 mRNA expression assay --- p.200 / Chapter 4.3.4 --- Ganoderma spp. As hormonal therapy --- p.201 / Chapter 4.4 --- DNA microarray analysis --- p.201 / Chapter 4.5 --- Further investigation --- p.204 / Chapter Chapter 5 --- Conclusion --- p.205 / Chapter Chapter 6 --- Reference --- p.207
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The effects of Tulbaghia violacea leaf, bulb and stalk extracts on Jurkat cells.Mackenzie, Jared Stuart. January 2012 (has links)
Studies have shown that the traditional healers have used Tulbaghia violacea (TV) (also
known as ‘wild garlic’) for the treatment of a number of ailments including fever,
tuberculosis, stomach problems, and oesophageal cancer. However, little is known with
regards to the anticancer and antiproliferative properties of this plant. Therefore, this
study investigated the effects of TV and domesticated garlic extracts on Jurkat cells, in
order to determine whether or not these extracts possess anti-proliferative properties.
Cultured Jurkat cells were treated with IC50 concentrations of garlic (14μg/ml), TV leaf
(256μg/ml), TV bulb (225μg/ml) and TV stalk (216μg/ml) extracts as determined by the
methylthiazol tetrazolium assay. Free radical production was measured using the
thiobarbituric acid reactive substance (TBARS) and nitric oxide (NO) assays, while
glutathione (GSH) concentration was measured using the GSH-Glo™ assay. The
apoptosis inducing properties of each extract were measured using flow cytometry
(Annexin V- Fluos and JC-1 assays) and luminometry (caspases 3/7, 8, 9 and ATP).
Western blots were run to determine protein expression, while comet and DNA
fragmentation assays were used to determine the level of DNA damage induced. Wild
and domesticated garlic extracts induced a significant increase in malondialdehyde
concentration ([MDA]), with TV bulb extract inducing the highest concentration
(p<0.0001). A significant increase in NO concentration was observed in the bulb
(p<0.0001) and stalk (p<0.001) extracts, and leaf (p<0.05) and stalk (p<0.05) TV
extracts significantly increasing GSH concentration. The longest comet tails were
observed in TV bulb extracts (p<0.0001) and comprised mainly of single strand breaks,
while the comets induced following garlic exposure contained double strand breaks. All
extracts, except TV leaf, increased the percentage of cells undergoing apoptosis.
Tulbaghia violacea leaf induced a significant (p<0.0001) increase in percentage of cells
undergoing necrosis, whereas TV bulb resulted in a significant (p<0.0001) decrease.
All TV extracts induced caspase 3/7 and 9 activity, with the most significant increase in
caspase 9 activity observed for TV leaf and bulb. No significant change in caspase 3/7
activity was evident for domesticated garlic. Cleavage of PARP and expression of
NF B and HSP 70 occured for all extracts. However, HSP 70 was not differentially
expressed. Exposure to wild and domesticated garlic extracts induced peroxidative lipid
and DNA damage within the cells, indicating oxidative stress. This damage occurred in
conjunction with increased percentage of cells undergoing apoptosis and expression of
caspase 3/7. Therefore, these findings suggest that TV is inducing cell death through
apoptosis in Jurkat cells using a number of mechanisms, including the induction of
oxidative stress. This is of clinical significance, as cell death through apoptosis is the
preferred method of action for anti-cancer drugs. / Thesis (M.Med.)-University of KwaZulu-Natal, Durban, 2012.
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Potential anticancer activity of in rhizomes of ginger species (Zingiberaceae family). / Potential anticancer activity in rhizomes of ginger species (Zingiberaceae family).Kirana, Chandra January 2003 (has links)
Title page, table of contents and abstract only. The complete thesis in print form is available from the University of Adelaide Library. / The aim of the work described in this thesis was initially to screen the ethanol extracts of eleven Indonesian ginger species (Zingiberaceae family) for anticancer activity. MCF-7 breast and HT-29 colon cancer cells were used for the investigations. Extracts of Zingiber aromaticum and Boesenbergia pandurata were found to be the most active species, similar to that of Curcuma langa which has been shown to possess anticancer activity in vitro and in vivo (Aruna and Sivaramakrishnan, 1992; Azuine and Bhide, 1992). These two active species were then further investigated. Bioactive compounds from the species were isolated and identified using various chromatography procedures and nuclear magnetic resonance (NMR) and their anticancer activities were further tested on MCF-7 breast and HT-29 colon cancer cells including cell cycle analysis and measurements of apoptosis. The ethanol extracts of these two active species were also investigated using the AOM-induced colon cancer model in rats. The antiinflammatory activity of the ethanol extract of Z. aromaticum was also investigated using dextran sulfate sodium (DSS) induced ulcerative colitis (UC) in rats. The inhibitory activity of ethanol extracts of rhizomes of 11 ginger species was initially tested against MCF-7 breast and HT-29 colon cancer cells using colorimetric tetrazolium salt (MTT) assay. Ethanol extracts of eight species (Amommum cardamomum, C. longa, C. mangga, C. xanthorrhiza, Boesenbergia pandurata, Zingiber aromaticum, Z. officinale, Z. cassumunar) showed a strong inhibitory effect on the growth of the cancer cells with the IC50 concentrations between 100-100 g/ml. The ethanol extract of Curcuma aeruginosa was less active (IC5o between 100-120 g/ml) and extracts of Kaempferia galangal and K. rotunda had no effect on the growth of either cell lines at concentrations up to 250 g/ml. Ethanol extract of C. longa was used as a comparison since curcumin, an active compound isolated from this species, has had demonstrated its anticancer activity in vitro, in vivo and is currently undergoing clinical trial against colon cancer (Greenwald, et al., 2001; Sharma et al., 2001). Extracts of Z. aromaticum and B. pandurata had very strong inhibitory activity similar to the extract of C. longa. Curcumin was not detectable in either Z. aromaticum or B. pandurata. The ethanol extracts of the active species were not toxic on human skin fibroblast cells (SF 3169). The ethanol extracts of Z. aromaticum and B. pandurata were further fractionated using two different solvents by reversed phase preparative HPLC. Fraction A was eluted with a mobile phase containing 5% vlv aqueous methanol containing 0.025% v/v trifluoroacetic acid (TFA) and fraction B was eluted with 100% methanol. The inhibitory activity of fractions was then investigated against HT-29 colon cancer cells and assayed using the MTT assay. Zerumbone, a sesquiterpenoid compound was isolated from fraction B of the extract of Z. aromaticum and a chalcone derivative, panduratin A was isolated from fraction B of the extract of B. pandurata. Curcumin was in fraction A of extract of C. longa. The anticancer activity of zerumbone and panduratin A was investigated using MCF-7 breast. HT-29 and CaCo-2 colon cancer cells. The inhibitory activity of the active compounds was assessed using the MTT assay. The ICso of zerumbone in each of the cell lines was about 10 uM and of curcumin on HTU29 cells was 25 uM. The IC50 of panduratin A in HT-29 cells was 16 uM and in MCF-7 cells was 9 uM. Zerumbone and panduratin A showed antiproliferative effects by alteration of the DNA distribution in the cell cycle and induction of apoptosis. HT-29 cells treated with zerumbone at concentrations of 10 -25 uM or panduratin A at concentrations of 9 -65 uM for 24 h were stained with propidium iodide (PI) to determine cell cycle distribution and analysed using FACScan flow cytometry. The proportion of cells in the S phase was reduced from 18.7% in untreated cells to 10.2% in HT-29 cells after treatment with zerumbone at 10 uM to 3.1% at 25 uM. Cells in the G2 phase increased from 18.5% at 10 uM to 40% at a concentration of 25 uM. Panduratin A increased the proportion of cells in the GO/G1 phase from 33% of untreated cells to 71% after treatment with 65 uM for 24 h. Panduratin A slightly reduced the proportion of cells in S phase and cells in G2/M phase also decreased from 36,8% in untreated cells to 15.4% at 65 M. Apoptosis was determined using double labelled (Annexin-V-Fluos and PI) and then evaluated using FACScan Flow Cytometry. Morphological features of apoptosis were also examined using DiffQuick stain and fluorescent Hoechst 3355 and 4,6-diamino-2-phenylindole (DAPI). Zerumbone induced apoptosis in HT-29 cells in a dose dependent rnanner, At 48 h, 2% of cells treated with 10 M of zerumbone underwent apoptosis, which increased to 8% when treated with 50 M, Panduratin A at 28 M increased the number of cells undergoing apoptosis from 2,2% to 16.7% when treated with a concentration of 65 M. The ethanolic extracts of Z. aromaticum and B. pandurata were also investigated using the azoxymethane (AOM) induced aberrant crypt foci (ACF) model of colon cancer in rats in a short and long term study. Ethanolic extracts of C. tonga and curcumin were used as comparison. The basal diet used throughout all animal studies in this thesis was a semi-purified AIN-93 G diet (Reeves et aI., 1993). ACF were induced by two doses (15 mg/kg BW) subcutaneously of AOM one week apart and ACF were visualised in the formalin fixed colon using methylene blue stain. The ACF study was run over a short (5 weeks) and long (13 weeks) experiments. Diets containing ethanol extracts prepared from the equivalent of 2% (w/w) dried rhizome of Z. aromaticum, B. pandurate or C. tonga in a short term study did not affect the formation of ACF in rats compared to those in the control diet group. The ACF formation in a short term study was dominated by small numbers of aberrant crypts (1 or 2) per focus. It is suggested that large ACF (4 or more ACs/focus) are better predictors of colon cancer (Uchida et aI., 1997; Jenab et aI., 2001). Diets containing ethanol extracts of the equivalent of 4% by weight of dried rhizomes of Z. aromaticum, B. pandurata, C. longa were investigated over 13 week study, Total ACF were significantly reduced by Z. aromaticum extract (0.34%) in the diet (down 21%, p<0.05) relative to rats fed the control diet. A similar reduction was observed with C, longa extract (0.86%) in the diet (down 24%, p<0.01) and with 2000 ppm curcumin. There was no significant different in small ACFs (1-2 ACs/ focus) between dietary treatments. The number of foci containing 3-4 ACs/focus was significantly reduced (35%, p<0,001) in animals fed the Z. aromaticum extract and 34% (p<0.001) of animals fed the C. tonga extract. The total number of ACF containing 5 or more ACs per focus of animals fed 0.34% Z. aromaticum extract was 41 % lower than control (p<0.05) and for 0.86 % C. tonga extract was 22% (not significant). A diet containing extract (0.56%) of B. pandurata did not significantly affect the formation of ACF compared to the control AIN group. The concentration of zerumbone in the Z.aromaticum extract diet was assayed at 300 ppm, and of curcumin in the C. tonga extract diet was also 300 ppm. The concentration of panduratin A was not assayed in the diet due to late identification of the active compound. The antiinflammatory activity of ethanol extract of Z. aromaticum was investigated using dextran sulfate sodium (DSS) induced ulcerative colitis in rats. Sulfasalazine, a widely used compound to treat inflammatory bowel disease (IBD) in humans was used as the positive control. Diets containing ethanol extracts (0.34% and 0.68%) prepared from the equivalent of 4% and 8% by weight of dried rhizomes of Z. aromaticum were given to the animals throughout the experiment. On day three, rats were given 2% DSS in drinking water for 5 d and then just water for 3 d and then were killed. During the DSS treatment rats were maintained in metabolic cages, body weight, food and fluid intake and clinical symptoms such as consistency of stools and blood in faeces were recorded daily. There was slight but not significant reduction in the body weight of rats fed 0.68% extract of Z. aromaticum in the diet due to reduced food consumption. The extract of Z. aromaticum (0.34%) and sulfasalazine suppressed clinical signs of ulcerative colitis. Eleven percent of the controls were hemoccult positive on day 2 after DSS administration, which progressed further by day three with 67% being hemoccult positive and 100 % on day five. By comparison, blood appeared on day 3 of rats treated with diet containing 0.34% and 0.68% extract of Z. aromaticum and 0.05% sulfasalazine, and only 33%, 67% and 22%, of rats being hemoccult positive on day 5 respectively. The disease activity index (DAI) of rats fed diet containing 0.34% extract of Z. aromaticum was about 0.4 and similar to those which were fed with diet containing sulfasalazine. The DAI of untreated rats was 1.4. The crypt score of rats fed the extract of Z. aromaticum was slightly reduced but it was not significantly different from those of untreated rats. Other histological scores were not significantly different between dietary treatments. Extract of Z. aromaticum significantly decreased the content of PGE-2 in colon tissue compared to that of untreated animals. There was a reduction of TX8-2 content in colonic tissue of rats fed with extracts of Z. aromaticum but this was not significant. The activity of myeloperoxidase (MPO) activity in the colonic tissue of rats fed with sulfasalazine was significantly lower than that of the untreated controls and those which fed with extracts of Z. aromaticum. The results from the studies performed in this thesis showed that extract of Z. aromaticum which contains an active sesquiterpenoid zerumbone have anticancer and antiinflammatory activity suggesting that the extract may have benefits as a chernopreventative agent. However further studies are needed to elucidate their other pharmacological actions. Panduratin A showed potential anticancer activity in cell culture in vitro. However an extract of B. pandurata did not have effect on the AOM-induced colon cancer model. Different cancer models such as breast and prostate cancer could be used to further investigate the anticancer activity of extract of B. pandurata and panduratin A and to elucidate their mechanism. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1097849 / Thesis (Ph.D.) -- University of Adelaide, Dept of Medicine, 2003
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The prevalence of dietary related complementary and alternative therapies and their usefulness among cancer patients attending the Colney Cancer Center in the Norwich Area, United KingdomVan Tonder, Esmarie 03 1900 (has links)
Thesis (MNutr (Human Nutrition))--Stellenbosch University, 2008. / Background: Cancer patients have been documented to use complementary and
alternative medicine (CAM) frequently, a subject that has been extensively researched.
There is however a lack in the current literature of controlled studies that investigate the
prevalence of CAM use among cancer patients compared to non-cancer controls.
Aim: To assess and compare the prevalence of dietary related CAM use among adult
cancer patients and non-cancer controls in the Norwich area, England.
Methods: Self-administered questionnaires were used to survey cancer patients
attending a comprehensive cancer centre in Norwich, and non-cancer controls attending
three dental surgeries also in the Norwich area. Questions addressed patient
demographics, information relating to cancer diagnosis (cancer cases only) and
information on CAM use. CAM users were asked about types and duration of CAM use,
reasons for use, information sources used, disclosure to health professionals, reported
side effects and benefits and satisfaction with CAM therapies.
Results: Questionnaires were distributed to 132 cancer cases and 126 controls, with 98
and 96 assessable replies received from the cases and controls respectively. Overall,
47% of the cancer cases used CAM, in comparison to 53% of the control group, with no
significant difference (p=0.673) between the two groups. Large quantities of juice,
multivitamins, fish oils and glucosamine were the most popular CAM therapies among
the two groups. Usage was significantly associated with the cancer site (p=0.036) and
duration of cancer diagnosis (p=0.050). Only 54% of the cancer cases and 44% of the
controls informed a health professional about their CAM use. The main reasons for
using CAM were to boost the immune system and to improve quality of life. Reported
benefits included increased optimism and hope.
Conclusions: Although CAM was commonly used by British cancer patients, there was
no significant difference in comparison to the non-cancer controls. Therefore, increased
awareness and knowledge of CAM use should not be limited only to those working with
oncology patients, but be extended to health professionals in all patient groups.
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