M.Sc. / Background: The continuous emergence of drug resistant strains of HIV as a result of errors made by reverse transcriptase coupled with undesirable side effects of available drugs, latency problems, cost etc, warrants the continuous search for new drug candidates. Chrysotherapy which is the use of gold compounds for the treatment of various ailments has been practiced since 2500 BC. The use of gold compounds such as auranofin for the treatment of rheumatoid arthritis has lead to remission of this disease. Gold compounds such as auranofin not only prevented the progression of arthritis but also increased the CD4+ count of an HIV positive patient who was not on antiretrovirals. These compounds have been implicated in the treatment of cancers, autoimmune diseases and microorganism infections. Objectives: In this work, novel gold compounds were evaluated with the aim of identifying lead compound(s) that can eventually serve as anti-HIV agents. Materials and Methods: Eleven gold (I) phosphine complexes, four of their corresponding ligands (compound without gold atom), and a gold (III) complex were tested for the ability to inhibit reverse transcriptase (RT) and protease (PR) in direct enzyme assays. Uptake of the compounds by host cells was evaluated with inductively coupled plasma atomic emission spectrometry (ICP-AES). Potential toxicity of the gold compounds was screened for by viability dyes and flow cytometry assays. To determine inhibition of whole virus by other mechanisms in addition to RT or PR, p24 production by infected cells was evaluated. Prior to all these analysis, stability of compounds in solution was determined by 31P nuclear magnetic resonance (NMR) and UV-visible spectroscopy. Results: The compounds were shown to be stable in solution over a one week period and were taken up by both continuous cell lines and primary cells. Eight of the gold compounds significantly inhibited HIV-1 reverse transcriptase at concentrations of 25 and 250 μM while four compounds and the four ligands did not. In a fluorogenic assay against HIV-1 PR, four of the gold compounds demonstrated inhibitory activity. The gold compounds were toxic to cells lines but not to primary cells. One of the complexes (EK231) significantly reduced p24 (p=0.0042) production at a concentration of 25 μM. Conclusion: Data provided here suggests that the therapeutic benefits of these gold containing compounds as potential HIV-1 reverse transcriptase and protease inhibitors should be considered.

Prof. H. Abrahamse

M.Tech. / The homoeopathic medicine Arnica montana is often prescribed in cases of trauma, before and after surgery and in cases where there is bleeding. Many conventional medical practitioners, however, do not advise its use for the above complaints due to the herbally prepared Arnica montana mother tincture containing coumarin derivatives which are said to have an anti-coagulant effect and cause a potential risk of bleeding. The aim of this particular study was to investigate the in vivo effect of the complex remedy Arnica montana composing of potencies 6C, 30C and 200C on coagulation and bleeding. This study forms part of a three part in vivo study to determine the effect of homoeopathic Arnica montana in various potencies on blood coagulation. The effect of Arnica montana on blood coagulation was evaluated by measuring the Bleeding Time (BT), activated Partial Thromboplastin Time (aPTT) and Prothrombin Time (PT). This is a double blind, placebo controlled trial with a total sample group of eighty healthy participants between the ages of eighteen to thirty five. As this study forms part of a three part study the total sample group was shared. Twenty participants were allocated to the placebo group and received 20% ethanol. Twenty participants were allocated to the experimental group and received the complex homoeopathic preparation of Arnica montana 6C, 30C and 200C in 20% ethanol. The Bleeding Time was measured by a trained medical technologist and blood samples underwent coagulation tests comprising of aPTT and PT. The study was conducted over a period of two weeks at the UJ Doornfontein Campus Homoeopathy Health Centre. After two weeks another venous sample was drawn by the phlebotomist and sent away for the same coagulation studies as described above. The technologist again measured the Bleeding Time. Results obtained from the Prothrombin Time, activated Partial Thromboplastin Time and Bleeding Time tests pre- and post medication were compared and v analysed. Analysis of data was done using SPSS 15.0 (Statistical Package for Social Sciences). Results showed that complex remedy Arnica montana 6C, 30C and 200C had no significant effect on blood coagulation and bleeding in vivo.

M.Tech. / Low level laser therapy, commonly known as LLLT or biomodulation, is a form of phototherapy which involves the application of low power monochromatic and coherent light to injuries and lesions to stimulate healing. In the medical field, lasers are classified as high power or surgical lasers and low level lasers which are used to stimulate cellular responses. Phototherapy has been successfully used for pain attenuation and induction of wound healing in non healing defects. Even though phototherapy has been found to be beneficial in a wide variety of therapeutic applications, it has been shown that phototherapy can induce DNA damage; however this damage appears to be repairable (Houreld and Abrahamse, 2008). DNA repair is vital to cells to avoid mutation. Literature reports show that red light or phototherapy up or down regulates genes involved in DNA repair (Zhang et al., 2003). N-methylpurine DNA glycosylase (MPG) is involved in DNA repair by catalysing the excision of a variety of modified bases. The exact mechanism by which phototherapy works is still poorly understood. Several authors have demonstrated that phototherapy enhances cell proliferation and migration. However, these cellular responses seem to confuse scientists as to whether wound healing is due to cell proliferation or migration or both. To determine the effect of phototherapy on cell proliferation or migration, a mini project was conducted (Zungu et al., 2008). Thus, cell proliferation was arrested using 5 mM hydroxyurea (HU) which is an antiproliferative drug. Wounded (W) human skin fibroblast cells (WS1, ATCC iii CRL 1502) were irradiated with 5 J/cm2 using a Helium-Neon (He-Ne) laser with a wavelength (λ) of 632.8 nm on day 1 and 4. Cell morphology, viability and proliferation were measured 24 h post irradiation. Reports indicate that several cell culture studies have used HU to control proliferation (Cai et al., 2000; Hamuro et al., 2002). Thereafter, the main study which was aimed at determining the effects of phototherapy on DNA damage and gene activation related to repair using 5 or 16 J/cm2 on W human skin fibroblast (WS1) cells was performed. Both studies involved growing WS1 cells aseptically in complete minimum essential medium (MEM) with Earle’s balanced salt solution and incubated at 37 °C in 5% CO2 and 85% humidity. Normal (N) and W cell cultures were irradiated with 5 or 16 J/cm2 30 min and 72 h (day 1 and 4) post wounding. Non irradiated cells (0 J/cm2) served as controls, while irradiated cells were the experimental groups. A wound was simulated by creating a central scratch across a monolayer of cells using a sterile 1 ml pipette. A 3 mW/cm2 He-Ne laser, λ 632.8 nm, was used to irradiate cells. After a repair time of 1 or 24 h on day 4, cell morphology (microscopy), cell viability (Trypan blue exclusion test and ATP luminescent assay), proliferation (XTT assay) and DNA integrity (alkaline comet assay with and without Formamidopyrimidine glycosylase [Fpg]) were assessed. The up or down regulation of the DNA repair gene, MPG, and regulation of three reference genes namely; beta Actin (ACTB), Glyceraldehyde 3 phosphate dehydrogenase (GPDH) and Ubiquitin c (UBC) were assessed by real time reverse transcriptase polymerase chain reaction (real time RT-PCR). iv Non irradiated HU treated cells had a reduced number of cells in the central scratch compared to non irradiated non treated cells, suggesting that HU inhibited cellular proliferation. Irradiated HU treated cells showed an increased number of cells in the central scratch compared to non irradiated treated cells. This observation proved that this increase was due to the stimulatory effect of irradiation with 5 J/cm2. The addition of HU had no significant effect on cell viability. The Trypan blue exclusion test showed no significant difference in percent viability between treated and non treated cells. Irradiated non treated cells showed a significant increase in the formazan dye, which is as a result of cleavage of XTT by the mitochondrial succinate dehydrogenase in actively proliferating cells, compared to non irradiated non treated cells (P=0.01). W cells, which were not irradiated, showed incomplete wound closure at both 1 and 24 h, while W cells irradiated with 5 J/cm2 showed complete wound closure. Similarly, W cells irradiated with 16 J/cm2 showed incomplete wound closure at 1 and 24 h. Cell viability, proliferation and DNA integrity assays showed that irradiated and non irradiated N cells were not significantly affected at both 1 and 24 h post irradiation. W cells (1 h) irradiated with 5 J/cm2 showed a significant increase in percentage cell viability and ATP compared to non irradiated W cells (1 h), (P=0.05 and P=0.04 respectively), while irradiation with 16 J/cm2 showed a significant decrease (P=0.014 and P=0.02 respectively). W cells (24 h) irradiated with 5 J/cm2 also showed a significant increase in percentage cell viability and ATP when compared to non irradiated W cells (24 h), (P=0.006 and P=0.04 respectively). Contrary, irradiation with 16 J/cm2 showed a significant decrease (P<0.001 and P=0.003 respectively). v Cell proliferation results showed that irradiation with 5 J/cm2 was stimulatory while 16 J/cm2 was inhibitory. The comet assay demonstrated that N cells irradiated with 5 or 16 J/cm2 exhibited an insignificant change in DNA damage at both 1 and 24 h when compared to their respective controls. This finding is in agreement with Karu et al., (2003) who observed that phototherapy does not alter the biological activity of cells which at the time of irradiation are functioning normally. W cells (1 and 24 h) irradiated with 16 J/cm2 showed a significant increase in DNA damage compared to their respective controls. However, there was a significant decrease in damage at 24 h compared to 1 h incubation due to the activation of DNA repair mechanisms. Though not significant, comet assay with Fpg (modified comet assay) showed more DNA damage compared to comet assay without the enzyme (conventional comet assay). It can be explained that the modified comet assay detected and cleaved oxidised bases in addition to single strand breaks, which the conventional comet assay detected, suggesting that the modified comet assay is more sensitive than the conventional comet assay. After validation of the three reference genes, ACTB was chosen to be the gene with which to normalise MPG expression in WS1 cells. It was found to be the least variable; its expression was consistent in W cells as well as cells exposed to a He-Ne laser at a fluence of 5 or 16 J/cm2. It produced an acceptable correlation coefficient (R2 >0.999) and PCR efficiency (94%). Conversely, other primers like GAPDH produced a low PCR efficiency (82%), while UBC produced a low R2 (0.898). Wang et al., (2006) recommends the value of R2 to be more than 0.995 and a PCR efficiency of between 90 and 100% for PCR results to be reliable. Other researchers have not supported the use of ACTB as a reference gene, stating that it is highly regulated (Wang et al., 2006), however this study showed that ACTB was not regulated by laser irradiation (632.8 nm at 5 or 16 J/cm2). The cell culture conditions and vi laser irradiation in this study did not induce MPG expression; perhaps an alternative repair pathway might have been induced, and hence repaired the DNA damage. In conclusion, the mini project demonstrated that HU is able to inhibit cell proliferation through its cytostatic effect without affecting the viability of W WS1 cells. This study also showed that irradiation of W cells with 5 J/cm2 using the correct parameters enhances cell migration and proliferation as evidenced by the presence of more cells in the central scratch in HU treated cells, and a significant increase in cell proliferation as shown by the XTT assay in non treated cells respectively. Thus, migration and proliferation are the direct result of phototherapy as both are involved in wound closure. This study further confirmed that irradiation of W cells with 5 J/cm2 stimulated ATP production, and hence cellular viability, as well as cell proliferation and migration. Irradiation of cells with higher fluences such as 16 J/cm2 is damaging to DNA and inhibitory to cell proliferation, migration and possibly to MPG expression. The study further showed that N cells are not stimulated by phototherapy, supporting the notion that lasers stimulate compromised cells. Thus, if they are growing normally there is nothing to stimulate. This understanding helps to clarify why N cells irradiated with 5 or 16 J/cm2 had insignificant responses. Cell culture conditions, fluence and duration of exposures are important parameters that can affect gene expression, and hence documentation of all experimental conditions needs to be emphasised and published if reproducibility is to be achieved.

M.Tech. / Low Level Laser Therapy (LLLT), also known as photo-biostimulation or simply phototherapy, has widely been used in the treatment of wounds, with its history dating back to the early 1960s (Ohshiro and Calderhead, 1991). Despite some literature reporting negative and non-existent cellular responses to LLLT, a growing body of literature reports the positive and beneficial effects of LLLT. LLLT has proved to be efficient in speeding and improving the quality of wound healing. Stressed cells respond more favourably to LLLT by recovering to their most natural state and functional capability (Bernett, 1998; Karu, 1998). When healing appears to be impaired, these tissues respond positively to the appropriate doses of light, especially light that is within 600 to 1,000 nm wavelengths (Enwemeka et al., 2004). Cellular responses to LLLT include changes in mitochondrial intracellular calcium ion (Ca2+) levels, Mitochondrial Membrane Potential (MMP), Adenine Triphosphate (ATP) concentration, and cyclic 5’, 3’ Adenosine Monophosphate (cAMP) (Karu, 1998). The mitochondrion is the power house of a cell and the major location of cellular ATP synthesis (Bayens and Dominiczak, 1999). ATP is an energy rich molecule that drives processes responsible for cell growth or proliferation (Klug et al., 2003). LLLT alters intracellular pH which is related to activation of ATPase leading to an increase in ATP production in the mitochondria of the cell (Alexandratou et al., 2002; Karu, 1998). However the mechanisms by which the beneficial effects are attained by cells in stress or injury state are not clear.

M. Tech. / Stem cells are defined as undifferentiated cells that can proliferate indefinitely and have the capacity of both self-renewal and differentiation to one or more types of specialised cells. Traumatic tissue injury and age-related degenerative diseases are a major problem in South Africa and worldwide. Stem cells could be used for tissue engineering and reconstructive surgery. In treating these conditions, the main principle of stem cell therapy is the replacement of damaged and dead cells in injured tissues and organs with new healthy ones expanded in vitro from stem cells (Orlic et al., 2002). These cells can be isolated from adipose tissue in significant numbers and exhibit stable growth and proliferation kinetics in culture and could be differentiated into bone, fat, cartilage and muscle when treated with established lineage-specific factors (Zuk et al., 2002). Low Level Laser Therapy (LLLT) is currently applied in the treatment of numerous diseases and pathological conditions (Gasparyan et al., 2004). LLLT produces positive effects on irradiated cells and tissues such as proliferation of cells, capillary growth and adenosine triphosphate (ATP) activation (Schindl et al., 1998). Low level laser radiation at different intensities has been shown to stimulate as well as to inhibit cellular processes (Moore et al., 2005). Epidermal growth factor (EGF) is a growth factor that plays important roles in the regulation of cell growth, proliferation and differentiation. This study investigated the effect of low level laser radiation alone as well as in combination with EGF on adult adipose derived stem cells (ADSCs) isolated from human adipose tissue. ADSCs were isolated from human adipose tissue through collagenase digestion and cultured in DMEM-F12 containing 10% FBS and antibiotics and incubated at 37°C in a humidified atmosphere of 5% CO2 (Zuk et al., 2001). iii Semi-confluent monolayers of ADSCs were exposed to low level laser at 5 J/cm2 using 636 nm diode laser with a power density of 12.1 mW/cm2 at room temperature in the dark. Cell morphology was monitored at 0, 24 and 48 h using an inverted light inverted microscope. Cell viability was evaluated at 0, 24 and 48 h using the Trypan Blue exclusion test and an adenosine triphosphate (ATP) luminescence assay. bFGF (basic fibroblast growth factor) indirect ELISA and optical density assays were used to monitor cell proliferation at 0, 24 and 48 h post irradiation. In addition the expressions of stem cell markers, β1-integrin and Thy-1, were monitored by immunocytochemical live cell surface labelling and Western blot analysis. Cells were incubated with EGF to enhance proliferation and differentiation and the cell morphology, viability and proliferation were monitored as well as the expressions of stem cell markers, β1-integrin and Thy-1. Morphology of the cells was not altered by irradiating them with 5 J/cm2 using diode laser at 0, 24 and 48 h. Cell viability and proliferation showed an increase at 24 and 48 h post irradiation. At 0 h, there was no significant difference between irradiated and non-irradiated cells in cell viability and proliferation. There was an increase in the expression of β1-integrin and Thy-1 after irradiation as shown by Western blot analysis and immunocytochemical live cell surface labelling. Cell viability and proliferation showed a significant increase at all time points post irradiation with the addition of EGF. There was no noticeable change in cellular morphology at any time point. Low level laser irradiation of human ADSC’s at 636 nm with 5 J/cm2 and 12.1 mW/cm2 increased the viability and proliferation of these cells in vitro. Furthermore, low level laser irradiation appeared to increase the expression of stem cell markers, β1-integrin and Thy-1. In addition, laser irradiation did not alter the morphology of the cultured cells. The addition of EGF to the cells also increased their viability and proliferation as well the expression of the markers, β1-integrin and Thy-1. The study showed that laser irradiation stimulates two important cellular responses namely cell viability and proliferation which indicates that ADSCs may be suitable for tissue engineering and future cell differentiation studies.

M.Tech. / Stem cells possess self-renewal capacity, long-term viability, and multilineage potential. Stem cells play important roles in normal physiological and disease processes, they also have great therapeutic potential. However, there have been controversies surrounding stem cells in political, religious and ethical arenas. Although the use of certain stem cells (i.e. embryonic stem cells) and the means by which they are obtained contravene certain basic ethical laws, researchers have developed methods with which to ethically obtain and create stem cell lines. Stem cells can be classified as either: totipotent, pluripotent, multipotent, oligopotent and unipotent (Moore, 2007). Totipotent cells have the ability to differentiate into all cell types of an embryo, including the extra-embryonic and post embryonic tissues and organs. Pluripotent cells have the potential to differentiate into almost all tissues found in an embryo (including germ cells), but are not capable of giving rise to supporting cells and tissues. Multipotent stem cells have progeny of several differentiated cell types - but all within a particular tissue, organ, or physiological system. A good example of multipotent cells, are the haematopoietic stem cells that produce blood cell-restricted progenitors, as well as all cell types and elements, such as platelets, that are normal components of blood. Oligopotent stem cells produce two or more lineages within a specific tissue, such as neural stem cells that are able to produce subsets of neurons in the brain. Unipotent cells self-renew, as well as give rise to a single mature cell type, a prime example being the spermatogonial stem cells, that give rise to spermatozoa (Moore, 2007). Adult human subcutaneous adipose tissue contains cells with multilineage developmental plasticity like marrow-derived mesenchymal stem cells (Strem et al., 2005, Tong et al., 2000). Adipose derived stem cells can be obtained in abundance and can differentiate into osteogenic, adipogenic, myogenic and chondrogenic lineages when treated with appropriate growth factors.

M.Tech. (Homoeopathy) / Insulin resistance is defined as the impaired ability of plasma insulin to facilitate peripheral glucose disposal, suppress hepatic gluconeogenesis and inhibit very low density lipoprotein (VLDL) output (Caceres et al., 2008). Insulin resistance is not a disease, but rather a feature and attribute of the Metabolic Syndrome, which is associated with a high risk of developing type two diabetes mellitus and cardiovascular disease (Brewer, 2005). Insulin resistance (IR) is globally regarded as the common and fundamental aetiological factor of the various components of the Metabolic Syndrome namely abdominal obesity, dyslipidaemia, insulin resistance and hypertension (Zimmet, 1991; Haffner et al., 1992 Stern, 1997; Beck Nielson & Groop, 1994; Tsai et al., 2012). Diabetes mellitus type 2 is a rapidly growing worldwide epidemic, but only diagnosed once the underlying metabolic abnormalities have caused damage. The fact that many recently diagnosed diabetic patients already suffer from so called “late complications of diabetes” indicates that the pre-diabetic condition is harmful to health and needs to be addressed promptly to slow down or avoid the progression to diabetes mellitus type 2 (Beck Nielson & Groop, 1994; Brewer, 2005; Tsai et al., 2012). Treatment of insulin resistance proves very difficult as dietary and lifestyle choices play an integral role in the development, treatment and management of insulin resistance; and insulin resistant patients also seem resistant to changing their behaviour (Brewer, 2005). Current conventional treatment options are limited in efficacy and may be associated with significant side-effects (Brewer, 2005; Snyman, 2009; Neal, 2003), while coherent studies on combination complementary forms of treatment are lacking (Chen et al., 2003; Guan et al., 2000; Hull, 2008; Verma et al., 1998; Winston & Kuhn, 2007; Ye et al., 2001). This study aimed to determine the efficacy of the herbal and nutritional formulation Glucostate™ in FoodState® form, on the HOMA index of insulin resistant patients. This was a collaborative randomized double-blind and double-dummy placebo controlled quantitative research study that included 40 participants. Due to the inherent nature of associated race, age and gender bias, participants were matched according to these criteria and randomly allocated to either an experimental or control group (Appendix C). The placebo group was shared between two collaborative studies. Participants volunteered to participate in the study, were between the ages of 20-45 years and consented to the procedures of the study. Participants in the treatment group received Glucostate™ tablets and placebo drops and the participants in the placebo group received placebo tablets and placebo drops. The research study was conducted over a period of 16 weeks per participant at the University of Johannesburg Health Training Centre, Doornfontein Campus. Participants were asked to maintain their original lifestyle and diet and continue as is normal for them, as alterations in weight have an effect on insulin levels. Participants were screened using blood pressure, abdominal girth, height, weight and Body Mass Index (BMI); these measurements were repeated at weeks 4, 8, 12 and 16. Fasting blood tests consisting of a lipogram, fasting glucose and fasting insulin level were done prior to and at the conclusion of the study. The Homeostasis Model Assessment (HOMA) index was calculated from the fasting insulin and glucose values. Results acquired from the research study were statistically analyzed by Statkon at the University of Johannesburg by means of descriptive statistics, parametric and non-parametric tests. The only parameters which showed statistically significant improvement for the Glucostate™ group and not the placebo group were systolic blood pressure (SBP) (p=0.004) and diastolic blood pressure (DBP) (p=0.050). There was no statistically significant change in any of the other parameters when compared to placebo. This research study determined that Glucostate™ was not effective in reducing insulin resistance and the parameters directly associated with its measurement especially when compared to the effects of placebo.

M.Tech. / Obesity is a serious health problem throughout the world, with the number of cases having increased three-fold over the last two decades, reaching epidemic levels in the United States (Kumar and Clark, 2009). According to the South African Demographic and Health Survey (SADHS) of 2002, 29.2% of South African men were overweight, whereas 56.6% of women were overweight. Obesity leads to a decreased quality of life and can have a detrimental effect on general well being (SADHS, 2002). Overweight and obesity have major psychological effects on the individual. Obese individuals often suffer from body image issues, unhappiness, a decreased morale, low self esteem and eating disorders (Rogge et al., 2004). The stigma associated with adolescent obesity causes bullying in school as well as childhood psychiatric morbidity (Gortmaker et al., 1993). The aim of this study was to determine the effect of an herbal formulation consisting of Coleus Forskohlii, Camellia sinensis, Coffea canephora, Caffeine, Evodiamine, Ilex paraguariensis and Phaseolus vulgaris on general well being in overweight and obese individuals. This study was a quantitative double-blind, placebo-controlled study, using matched pairs according to gender and body mass index (BMI). The study was conducted over twelve weeks during the period of October 2011 to January 2012 at the University of Johannesburg’s Homeopathy Clinic. The conducted study formed part of a larger study where the research sample was shared by two additional researchers (Durrheim et al., 2012) (Withers et al., 2012). This allowed for a larger sample group to be tested with numerous variables being researched. The study included sixty overweight and obese males and females between the ages of eighteen and forty five, with a BMI above 25 kg/m² and under 35kg/m². Participants were required to sign a Participant Information and Consent form (Appendix B) giving the necessary information regarding this study. The groups were randomly matched assigned by the dispenser (according to the participant’s BMI) to the experimental group or placebo group, each consisting of thirty participants. The medication was randomised by the homeopathic dispenser at the University of Johannesburg’s Homeopathy Clinic. The experimental group received a bottle of capsules containing the herbal formulation, and the placebo received identical looking placebo capsules containing a pharmaceutical maize starch.

Until now, emerging viral diseases have been posing ongoing threats to the global public health. Among the notorious viruses, the HIV that causes the AIDS has been spreading continuously since it was first identified in 1981 and is the most quickly spreading disease of the century. Although considerable advance has been made by drug discovery groups, the therapeutic management is still challenged by the rapid mutations of the virus to yield resistant strains, so as the emergence of side effects. Therefore, the development of novel potent anti-HIV agents is urgently sought. Owing to the chemical diversity, we believe that natural products may serve as potential "lead" compounds for discovery of anti-HIV drugs.;In order to search for novel naturally occurring compounds with potent inhibitory effects against HIV, we began with isolation of natural products from two medicinal plants of Justicia by means of silica gel column chromatography, and preparative HPLC, namely, J. gendarussa that displayed potent anti-HIV activity in our initial screening, and J. procumbens, and their chemical structures and determinated by spectroscopic and chemical methods such as IR, UV, HRESIMS, 1H NMR and 13C NMR spectrometry (including DEPT, 1H-1H COSY, and HMBC techniques). Upon the complete identification of compounds, we focused on the synthesis of one potential lead compound isolated from J. gendarussa, patentiflorin A (3). Nevertheless, we evaluated all the isolated natural compounds and synthetic 3 via bioactivity screening for anti-HIV activity.;In the phytochemical investigation of J. gendarussa, a rare, shade-loving, quick-growing, evergreen scented shrub collected in Vietnam, the bioassay-directed fractionation of the methanol extract of the roots and stems of the plant led to the isolation two new arylnaphthalide lignan glycosides, named justiprocumins A and B (1--2), together with a known one, patentiflorin A (3). On the other hand, the phytochemical investigation of the methanol extract of the aerial parts of J. procumbens resulted in the isolation of four novel arylnaphthalide lignans, procumbenosides G (4), H (5), I (6) and J (7), along with 23 known compounds, namely, tuberculatin (8), procumbenoside B (9), procumbenoside E (10), ciliatoside B (11), ciliatoside A (12), 5-methoxy-4, 42-di-O-methylsecolariciresinol (13), secoisolariciresinol dimethyl ether (14), 2, 3-bis(3, 4-dimethoxybenzyl)-4-hydroxybutyl acetate (15), secoisolariciresinol (16), hemiariensin (17), ariensin (18), secoisolariciresinol dimethyl ether diacetate (19), hinokinin (20), justicidin E (21), justicidin D (22), justicidin C (23), cilinaphthalide A (24), 5'-methoxy-4'-O-methyllariciresinol (25), 3, 5, 7, 32, 42, 52-hexamethoxyflavone (26), 3, 5, 7, 8, 32, 42-heptamethoxyflavone (27), 3, 5, 7, 8, 32, 42, 52-heptamethoxyflavone (28), methyl ferulate (29) and loliolide (30). In addition, the compound 3 was totally synthesized with a yield of 68.3%;In the anti-HIV evaluation for all the isolated compounds using the defective HIV-based pseudotyped assay, patentiflorin A (3) was found to have anti-HIV activity with an IC50 value of 26.9 nM, while justicin E (21) showed 65.4 % inhibitory effect against HIV replication at 2.5 μg/mL. In the evaluation for the broadness of the spectrum of anti-HIV activity using a standardized human PBMC assay, 2 gave IC50 values of 14-21, and 3 gave IC50 values 24-37 nM in inhibiting the particle production of all the four HIV-1 isolates [BAL and SF162 (both are M-tropic), LAV0.04 (T-tropic), and 89.6 (dual tropic)], while the synthetic 3 showed quite similar activity as that of natural 3. In the test of cytotoxicity, natural 3 exhibited no apparent cytotoxicity at 19.0 mM in A549 and Hela cells, and the synthetic 3 displayed much lower cytotoxicity (CC50: 75.5 mM) than that of the natural 3 (CC50: 18.4 mM) in PBMC cells. That means 2 and 3 have great potentials as anti-HIV lead compounds for further drug development.;In conclusion, natural compounds isolated from medicinal plants serves as one of the most important sources of potentially anti-HIV compounds, which can be employed as "lead" compounds to develop novel therapeutic drugs against HIV.