Lo, Yu Ching. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 79-82). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abbreviations --- p.ii / Abstract --- p.iii / Table of Contents --- p.v / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Basic Theory of FTIR --- p.1 / Chapter 1.1.1 --- Infrared Spectroscopy --- p.1 / Chapter 1.1.2 --- Dispersive Infrared Spectrometer --- p.2 / Chapter 1.1.3 --- Fourier Transform Infrared Spectrometer (FTIR) --- p.3 / Chapter 1.1.4 --- Advantages of FTIR --- p.5 / Chapter 1.1.4.1 --- Multiplex advantage --- p.5 / Chapter 1.1.4.2 --- Throughput advantage --- p.5 / Chapter 1.2 --- Traditional Chinese Medicine (TCM) --- p.6 / Chapter 1.3 --- Identification of Traditional Chinese Herbal Medicine --- p.8 / Chapter 1.3.1 --- Traditional method for the identification of TCM --- p.8 / Chapter 1.3.2 --- Instrumental method for the identification of TCM --- p.9 / Chapter 1.3.2.1 --- Identification of TCM using fingerprint method --- p.9 / Chapter 1.3.3 --- Identification of TCM using FTIR fingerprint method --- p.10 / Chapter 1.4 --- Objective --- p.11 / Chapter Chapter 2 --- Experimental / Chapter 2.1 --- Outline of the Method --- p.12 / Chapter 2.2 --- Reagents and Glassware --- p.13 / Chapter 2.3 --- Instrumentation --- p.13 / Chapter 2.4 --- Library Search Program --- p.13 / Chapter 2.5 --- Samples --- p.14 / Chapter 2.6 --- Sample Pretreatment --- p.16 / Chapter 2.7 --- Extraction of Ingredients --- p.16 / Chapter 2.8 --- Preparation of KBr Pellet --- p.17 / Chapter 2.9 --- IR Spectrum Measurement --- p.17 / Chapter 2.10 --- Data Processing --- p.18 / Chapter 2.11 --- IR Databases --- p.18 / Chapter 2.12 --- Reproducibility of Extraction --- p.29 / Chapter Chapter 3 --- Application of Library Search Techniques - Results and Disscussion / Chapter 3.1 --- Introduction --- p.31 / Chapter 3.2 --- Euclidean Search Method --- p.32 / Chapter 3.2.1 --- Similarly score of reference spectra --- p.32 / Chapter 3.2.2 --- Similarity score of known sample spectra --- p.37 / Chapter 3.3 --- Soft Independent Modeling of Class Analogy (SIMCA) --- p.45 / Chapter 3.3.1 --- Verification Diagnostic Report of the Reference flowers --- p.47 / Chapter 3.3.2 --- Classification of Flowers --- p.51 / Chapter 3.4 --- Performance Limitation --- p.75 / Chapter 3.4.1 --- Euclidean Search Method --- p.75 / Chapter 3.4.2 --- SIMCA --- p.75 / Chapter 3.5 --- Conclusion --- p.77 / References --- p.79

Bioactivity-guided isolation of SFE oil led to the identification of another sesquiterpene lactone, 6-O-angeloylprenolin, containing the bioactive alpha, beta-unsaturated cyclopentenone. MTT results showed that CNE cells were more susceptible to 6-O-angeloylenolin than the normal Hs68 cells. Besides, the inhibitory effect of 6-O -angeloylenolin on the CNE cells was slightly stronger than that of cisplatin, the positive control, albeit statistical insignificance. / Both volatile oils prepared by supercritical fluid extraction (SFE) and steam distillation (SD) were evaluated for their anti-NPC potential. Results showed that SFE oil was much stronger than that of SD oil. SFE oil significantly inhibited the growth of CNE cells by dysfunctioning the mitochondria and activating caspases. Gas chromatography-mass spectrometry analysis revealed that the responsible principals in the SFE oil were likely homologues of sesquiterpene lactones. / Centipeda minima (L.) A. Br. (Compositae), a Chinese medicinal herb, is used to treat nasopharyngeal carcinoma (NPC) in the Chinese folk. However, there is a paucity of information on its anticancer activities. In particular, both of its anti-NPC potential and the potent constituents remain elusive. / In this study, the n-hexane fraction of C. minima showed broad spectrum of inhibitory effects on five human cancer cell lines, including the breast carcinoma MCF7 cells, the prostate carcinoma PC-3 cells, the hepatocellular carcinoma Hep G2 cells, the nasopharyngeal cancer CNE cells and the acute promyelocytic leukemia HL-60 cells, with IC 50 values ranging from 6.1 to 47.3 mug/mL. Bioactivity-guided separation of the n-hexane fraction using the CNE cells as the cellular system led to the isolation of a sesquiterpene lactone, 2beta-(isobutyryloxy)florilenalin (IF), which contained the bioactive alpha-methylene-gamma-lactone ring. IF significantly induced CNE cell death with an IC50 value of 3.1 mug/mL. Despite this potency, its effect on the normal Hs68 cells was much weaker, with an IC50 value larger than 50 mug/mL. Its inhibitory effect on the CNE cells ascribed to apoptotic induction as evidenced by the cumulation of sub-G1 cell population, DNA fragmentation and nuclear condensation, caspase-3 activation and poly (ADP-ribose) polymerase (PARP) cleavage. Mechanistic study showed that both extrinsic and intrinsic apoptotic pathways were activated. In the extrinsic pathway, IF activated caspase-8, which further induced the activation of caspase-3 and caspase-7. In the intrinsic pathway, IF regulated the expressions of Bcl-2 family proteins, followed by depletion of mitochondrial membrane potential (Delta&PSgr;m), the release of cytochrome c to cytosol, the activation of caspase-9 and other downstream caspases, and finally the induction of apoptosis. / Mechanistic investigation showed that 6-O-angeloylenolin caused cell cycle arrest at S and G2/M phases and induced apoptosis in CNE cells. For the cell cycle arrest, a sharp decrease was found in the expressions of cyclin D1, cyclin D3, cdc25c, and p-cdc25c, with concomitant decrease in CDK4, cyclin A, cyclin E, p-Rb(Ser780), p21Waf1/Cip1, cdc2 and p-cdc2. For the induction of apoptosis, externalization of phosphatidylserine and depletion of Delta&PSgr;m prior to the detection of sub-G1 peak were found. Other apoptotic features including the presence of apoptotic bodies, the activation of caspase-3 activity and the cleavage of PARP were observed. Activation of caspase-8 and caspase-10 was detected. Besides, 6-O -angeloylenolin induced the release of cytochrome c and AIF to cytosol. The former formed apoptosome with caspase-9, further activated the downstream caspase-3 and caspase-7 and cleaved PARP, while the latter was translocated into the nucleus and caused large-scale DNA fragmentation. Failure of the pan-caspase inhibitor, z-VAD-fmk, to interrupt the apoptotic induction by 6-O-angeloylenolin suggested that caspase-independent pathway was involved. 6-O-Angeloylenolin was able to activate Akt, ERK and JNK pathways. But only with the addition of JNK inhibitor (SP600125), significant suppression of the 6-O-angeloylenolin-induced apoptosis was observed, suggesting the involvement of the JNK pathway in the apoptotic pathway. Taken together, this study provided a better mechanistic insight into the potential application of 6-O-angeloylenolin as a candidate for NPC treatment. / Overall, this study revealed that two sesquiterpene lactones, including IF and 6-O-angeloylenolin were found to be responsible for the potent anti-NPC effect of C. minima. This study reiterates the notion that Chinese medicinal herbs traditionally applied to cancer treatment may be good sources of anticancer drug discovery, and sesquiterpene lactone may be a group of noteworthy lead compounds displaying anti-NPC potential. / Su, Miaoxian. / Adviser: Hau Yin Chung. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 100-113). / 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, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Thesis submitted to the Faculty of Science under the school of Molecular and Cell Biology, University of the Witwatersrand, in partial fulfilment of the requirements for PhD. Gauteng, Johannesburg, 2015 / Cancer is an enormous burden of disease, accounting for millions of deaths annually worldwide. Today, more people are dying from cancer than HIV/AIDS, malaria and tuberculosis combined. According to the American Cancer Society, it is expected that the global cancer burden will double by 2030 if preventative measures are not applied. Breast and gynaecological cancers remains a big scourge in developing countries, with breast cancer being the most common cancer and gynaecological cancers accounting for approximately 25% of all cancers in women in developing countries. Currently, the standard cancer treatments include surgery, radiation and chemotherapy. Adverse toxicities have been associated with these therapies and their effectiveness is also limited to drug resistance. The cost of treatment is another major burden. Limitations associated with these conventional cancer treatments have made discoveries of novel therapeutics which exhibit less toxicity and at a lowered cost of paramount importance. Medicinal plant extracts have recently attracted attention to modern medical science research with their non-lethal activity. Currently, up to 50% of the world drugs including chemotherapeutic drugs such as taxol and camptothecin are made from natural products or their derivatives. In this study we aimed to investigate the anti-cancer properties of the medicinal plant Euphorbia tirucalli. The crude extracts of E. tirucalli extracted using butanol; hexane and methanol solvents were screened for antiproliferative activity in breast (MCF-7 and MDA-MB231), ovarian (RMG-1) and cervical (SiHa) cancer cell lines. MTT assay and Real-Time Cell Analyzer (RTCA), xCELLigence were used to determine cytotoxicity of the extracts and calculate IC50. From MTT and xCELLigence results, we observed that E. tirucalli extracts exhibited dose-dependent inhibition of cell proliferation with RMG-1 and MCF-7 cells being more sensitive than MDA-MB231 and SiHa cells to all three extracts for an unclear reason. The butanol extract appeared to exhibit iv the most cytotoxicity with all cell lines reaching IC50 at low extract concentrations. Most therapies in anticancer treatment, such as chemotherapy, mainly induce cell death by causing either G0/G1 or G2/M cell cycle arrest and then inducing an apoptotic pathway. Therefore, cell cycle arrest and the induction of apoptosis in cancer cells become the major indicators of anticancer effects. Cells were stained with propidium iodide dye to determine if cells were arrested at G0/G1 or G2/M cell cycle stages while annexin V and PI staining were used to determine the type of cell death induced by the extracts. Cell cycle analyses revealed MCF-7, MDA-MB231 and SiHa cancer cells underwent arrest at G0/G1 following treatment with the plant extracts. Annexin V and PI staining revealed different proportions of apoptotic and necrotic populations. The extracts mainly induced apoptosis on MCF-7 and MDA-MB 231 cells, with the butanolic extracts inducing the most apoptosis. RMG-1 and SiHa cells had a high proportion of cells undergoing both late apoptosis and necrosis. The molecular mechanism of cell death induction was investigated using real time PCR and western blot. From the gene expression studies, p21 was observed to be over expressed in all cells following all treatments, in line with the observed cell cycle arrest at G0/G1. The extrinsic pathway of apoptosis was identified as the type of cell death induced with caspase 8 being overexpressed in MDA-MB 231 cells treated with butanol and hexane extracts. Upon further fractionation, flavonoids and especially isorhamnetin were identified as the active compounds in these extracts. Overall, the plant contains compounds that have some activity against cell proliferation and can be a promising tool to treat cancer cells. However, more work needs to be done to verify which compounds are mainly involved.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Traditional Chinese medicine (TCM) plays an important role in the Chinese healthcare system for over five thousand years. It includes the use of herbal medicine, acupuncture, Tui Na (推拿), and diet therapy. TCM helps to maintain a balance of Yin-Yang (阴阳), Five Phases (五行), Meridians (经络) and Qi (气) inside the body. In practise, pregnant women take tocolytic drugs to tonify the blood and qi to provide a continuous supply of nutrients for baby. Traditional Chinese herbal medicines usually prescribed as a complex formula to produce synergistic or agonistic effect to maintain a well balance of the above components in human bodies. Moreover, TCM usually cannot produce immediate effect on patients, therefore, the efficacy of individual component remains largely unknown. This study aims to investigate whether Chinese tocolytic drug components could modulate fertility by affecting the in vitro spheroid (blastocyte surrogate) attachment process by using trophoblastic (JEG-3) and endometrial epithelial (Ishikawa) cells to mimic the embryo-endometrial implantation process. Nine Chinese herbal medicinal compounds (Atractylenolide I(白术内酯), Atractylenolide II(白术内酯II), Atractylenolide III(白术内酯III), Paeoniflorin(芍药苷), Albiflorin(芍药内酯苷), Nuzhenide(女贞子甙), Ecliptasaponin A(旱莲甙A), Wedelolactone(蟛蜞菊内酯) and Columbianadin(二氢欧山芹醇当归酸酯)) which are commonly found in traditional Chinese tocolytic drug formula were selected to study (1) the toxicity of the drugs on trophoblastic (JEG-3) and endometrial epithelial (Ishikawa) cells growth, (2) the effect of three tocolytic drugs (Atractylenolide I, Atractylenolide II and Atractylenolide III) on spheroid attachment, and (3) their effect of the expression of Wingless (Wnt) signaling molecules (Active-β-Catenin, Axin-2, β-catenin, E-cadherin, GSK-3β, and Mucin-1). It was found that the nine compounds, Atractylenolide I, Atractylenolide II, Atractylenolide III, Paeoniflorin, Albiflorin, Nuzhenide, Ecliptasaponin A, Wedelolactone and Columbianadin did not affect cell viability at 25μM, 25μM, 5μM, 0.2μM, 125μM, 125μM, 125μM, 5μM and 25μM, respectively, by cell proliferation assay. However, at these concentrations, the spheroid attachment was not significantly increased by Atractylenolide I, Atractylenolide II and Atractylenolide III. Interestingly, the protein expression of GSK-3β and Active-β-catenin were up-regulated by the three compounds in both cells and JEG-3 cells respectively. The expressions of Axin-2 and E-cadherin were up-regulated by Atractylenolide III in Ishikawa cells and Atractylenolide II in JEG-3 cells. Atractylenolide I and Atractylenolide III increase the Ishikawa cells expression of Active-β-catenin and β-catenin respectively and together suppress the JEG-3 cells Mucin-1 and β-catenin expression. In conclusion, the nine tocolytic compounds have different effect on cell proliferation. Atractylenolide I, Atractylenolide II and Atractylenolide III did not enhance the attachment rate of JEG-3 spheroid onto Ishikawa monolayer. However, they affected Wnt-signaling molecules expression, suggesting that they may modulate endometrial receptivity. Further experiments are needed to study their combined effect on co-culture and expression of Wnt-signaling molecules. / published_or_final_version / Obstetrics and Gynaecology / Master / Master of Medical Sciences

M. Tech. Chemistry. / Discusses essential oils from indigenous Lippia species, L. scaberrima, L. rehmannii and L. javanica, can be used as bactericides and fungicides. Incorporation of these oils into a slow release gel will provide a mechanism for controlled prolonged release of the volatile compounds.

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

Antioxidant activity was measured in 14 plant species incorporated in more than 30% of Tibetan medicines used for cardiovascular disease and related symptoms according to indigenous pharmacopoeias. The study was undertaken in order to explore possible dietary/medicinal elements which may contribute to the reportedly low incidence of cardiovascular disease among Tibetan highlanders despite high hematocrit levels and a high saturated fat/low fruit and vegetable diet. Extracts of Terminalia chebula, Syzygium aromaticum, Aquilaria agallocha, Santalum album, Amomum subulatum, Justicia adhatoda and Myristica fragrans were strong scavengers of the 1,1 diphenyl-2-picryl-hydrazyl (DPPH) radical (P < 0.05). Cu2+-catalyzed low-density lipoprotein (LDL) oxidation was measured in vitro using thiobarbituric acid reactive substances (TBARS) formation and monitoring change in absorbency at 234 rim from conjugated dienes. The hexane fraction of S. aromaticum significantly reduced LDL susceptibility to oxidation (1339.96 +/- 7.01 min. lag time, P < 0.05), more than three times longer than TroloxRTM (431.02 +/- 21.19 min). Results of TBARS (90 min.: r = 0.71, P < 0.005; 180 min.: r = 0.74, P < 0.005) and DPPH (r = 0.69, P < 0.05) assays positively correlated to conjugated dienes formation. Our results suggest that these plants are likely to contribute to the therapeutic effects of traditional drugs used to treat cardiovascular disease.

This research study undertook to investigate and evaluate for efficacy and safety, the herbal remedies used for treating Diabetes mellitus in northern KwaZulu-Natal. In addition, it sought to gain knowledge and better understanding of traditional healing systems and the medicinal use of the natural flora. During the process of assimilating the desired information, the epidemiological and socio-economic factors which determine the form of medicine chosen by rural people in the region, were quantified. Both aspects of explanatory studies i.e. experimental and observational were used. Firstly, to evaluate the safety of the two herbal remedies, laboratory animals were given an oral dose of the herbal medicine and observed for a period of 14 days. Efficacy was assessed by treating Streptozotocin-induced diabetic rats with the herbal remedies and comparing their effect on blood glucose with that of a conventional sulphonylurea. The second part of the study was observational and it involved monitoring human subjects (patients) for twelve months, who were already taking the herbal preparations (n=56) and comparing their prognoses with that of a group taking conventional medicine (n=97). A third group using both types of medicine (n=42) was included as control measure for a possible confounding factor. Main outcome measures; Both subjective and objective measures of the perceived health of the diabetic patients were measured, as well as the determinants of using traditional medicine versus conventional medicine. The battery of toxicity tests which utilises behavioural and functional observations of the laboratory animals, yielded no signs of toxicity or abnormal behaviour. The histopathological examination results of the sample organs from the treated rats also revealed no signs of abnormality that could be attributed to the herbal remedies tested. There was no sex variation recorded in the response. The first HP tested (HP-1) demonstrated minimal hypoglycaemic effect whereas HP-2 significantly lowered the blood glucose of the streptozotocin-induced diabetic rats by an average of 59%. This was comparable to the conventional medicine (Glibenclamide) used in the experiment. After 12 months of follow-up, 93 % of traditional medicine users (n=56) were convinced that their blood sugar was controlled because of the traditional remedy they were using. The proportion of diabetic cases who used conventional medicine were no better off than those who used traditional medicine or vice versa. Health status and the financial situation (income) of the respondents greatly influenced their choice for diabetic treatment. The herbal remedies that were investigated were non-toxic and safe for use and internal consumption. One preparation demonstrated a significant hypoglycaemic effect, which was comparable to the conventional allopathic medicine used in treating Diabetes mellitus. This study should serve as a springboard to encourage more pharmacological evaluation of herbal medicines. / Thesis (Ph.D.)-University of Durban-Westville, 1998.

The search for cheap, non toxic and readily available antidiabetic drugs has been a challenge for researchers and the pharmaceutical industries. Diabetes mellitus is a metabolic disease characterized by defects in the synthesis of insulin and/or insensitivity to the action of insulin at the target cells. The disease has been on the increase mostly in developing countries where large proportions of the population have little access to good medical care due to either accessibility or non availability of synthetic drugs. This has led to the use of medicinal plants to treat diabetes because it is safe, cheap and with few side effects. There is little scientific evidence on the dosages, active compounds, mechanisms of action and toxicity of these traditionally used plants. Two of the most frequently used plants; Albuca setosa and Albuca bracteata were investigated in this study. The qualitative analysis of different extractions of these plants revealed the presence of phenolics, alkaloids, tannins and saponins. The antioxidant properties of aqueous, acetone and methanollic extracts of Albuca setosa and Albuca bracteata were investigated using models such as Diphenyl-1-Picrylhydrazyl (DPPH), 2, 2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), Ferric ion reducing antioxidant potential (FRAP), Nitric Oxide and Hydrogen Peroxide (H2O2). Both plants revealed inhibitions against DPPH in a concentration - dependent manner with Albuca setosa (0.330 mg/ml) showing higher activity than Albuca bracteata (0.647 mg/ml) determined from the IC50. The aqueous extract of Albuca setosa showed a higher inhibition against DPPH radical compared to the Albuca bracteata aqueous extract at all concentrations investigated. The isolated saponins from Albuca bracteata had a higher DPPH scavenging activity than the crude methanolic extract of the plant in a concentration - dependent manner but are significantly different from each other at 0.4, 0.6 and 1.0 mg/ml only. The IC50 of the saponins was also observed to be higher than the crude extracts and standards.The Albuca setosa aqueous extract showed a higher percentage inhibition of ABTS radicals than Albuca bracteata at all the concentrations investigated. Overall, the Albuca setosa aqueous extract (0.0809 mg/ml) showed maximum activity against ABTS radicals. The iron reducing power was significantly higher (P < 0.05) in the methanolic extract of both plants compared to the aqueous counterpart. Overall, the Albuca bracteata aqueous extract (0.344 mg/ml) showed maximum activity as indicated by the IC50. The aqueous extracts of both plants also revealed percentage inhibitions in a concentration - dependent manner against NO2. The aqueous extract of Albuca bracteata bulb was more active against nitric oxide and hydrogen peroxide inhibition. In this study, the cytotoxicity of the extracts was evaluated at a high dose of 100 μg/ml on Chang liver cells and determined using MTT, crystal violet, glucose consumption, lactate production and lactate dehydrogenase release and FRAP. The aqueous extracts of both Albuca setosa and Albuca bracteata were non-toxic on Chang liver cells at the concentrations investigated. The MTT revealed that the aqueous extract of Albuca setosa bulb had the optimum cell viability of 108.09 percent while the acetonic extract of Albuca bracteata showed the least cell viability (37.72 percent) compared with the control. The crystal violet test also revealed the acetone extract of Albuca bracteata to have the least percentage of cell viability at 31.47 percent, while the aqueous extract of Albuca setosa showed the maximum cell viability at 112.5 percent. The aqueous extracts of both plants showed higher percentage cell density on the second day of incubation from the proliferation assay. All the tested samples were observed to consume more glucose than the blank except for the methanollic and acetone extracts of Albuca bracteata bulb. The aqueous and methanolic extracts of Albuca setosa bulbs produced the highest lactate with 120.2 μg/ml and 113.7 μg/ml respectively. The acetone extracts of both Albuca setosa and Albuca bracteata revealed toxicity with a higher lactate dehydrogenase release compared to the control.