Traditional Chinese medicine (TCM) is a typical traditional medicine (TM) with a long-standing history of preventing and curing diseases in China and other countries in East Asia. Standardization of TCM has been a topic of discussion over the past few decades in China with the goal of promoting advances in TCM in China and elsewhere around the world. Many quality and safety control standards for TCMs have been implemented in China, but systematic standards of efficacy have not been established for TCMs until now because of the absence of evidence from clinical practice. Evidence-based medicine (EBM) is the best way to provide evidence from clinical practice, but the quality of current EBM studies of TCM, and especially randomized controlled trials (RCTs) of TCM, needs to be improved. International registration of clinical trials (CTs) of TCM is a good way to provide quality evidence from clinical practice of TCM because it can improve research transparency and ultimately enhance the validity and value of scientific evidence. This evidence will provide the springboard for efforts to standardize TCM.
Neurofibromatosis (NF) is a family of genetic diseases which are caused by dysfunction of either NF1 gene or NF2 gene. One in 3,000 people suffer from this tumor-carrying NF. NF1 gene product is a RAS GTPase activating protein (GAP) of 2,818 amino acids, which normally attenuates the GTP-dependent signal transducing activity of the G protein RAS. Dysfunction of this GAP leads to the abnormal activation of RAS, and eventually an oncogenic kinase called PAK1 as well. NF2 gene product is ''Merlin'' which directly inactivates PAK1. Thus, dysfunction of Merlin causes the abnormal activation of PAK1. In other words, dysfunction of NF1 gene (causing type 1 NF) is basically the same as dysfunction of NF2 gene (causing type 2 NF). In fact the growth of both NF1 and NF2 tumors requires PAK1, and all PAK1 blockers, synthetic chemicals or natural products, suppress the growth of these NF tumor cells both in vitro (cell culture) and in vivo (mice). However, until recently, no FDA-approved effective NF therapeutics is available on the market. Here a series of anti-PAK1 products shall be introduced, which would be potentially useful for the life-long treatment of NF patients in the future. These include the most potent HDAC (histone deacetylase) inhibitor FK228 (IC50: around 1 nM), that eventually blocks PAK1, the direct PAK1 inhibitor PF3758309 (IC50: around 10 nM), a CAPE (caffeic acid phenethyl ester)-based propolis extract called ''Bio 30'' from NZ (New Zealand), and an ARC (artepillin C)-based green propolis extract (GPE) from Brazil. Although the first two drugs are potent, none of them is available on the market as yet. The last two natural (bee-made) products are available on the market, and have been used for the therapy of NF and tuberous sclerosis (TSC) as well as many PAK1-dependent solid cancers such as breast and pancreatic cancers as well as glioma, which altogether represent more than 70% of all human cancers. Since PAK1 is not essential for the normal cell growth, propolis extracts cause no side effects.
Nucleocytoplasmic transport of proteins across the nuclear pore complex (NPC), mediated by the nuclear localization signal (NLS) and the nuclear export signal (NES), is a vital homeostatic process in eukaryotic cells and also in mitogen-activated protein kinase (MEK) signaling molecule in tumor cell proliferation. Some viruses, including the influenza virus and HIV-1, also employ this nuclear export mechanism during their life cycle. Hence, drugs that control nucleocytoplasmic transport of proteins are putative candidate antivirals or anti-cancer agents. Thus, we previously developed a GFP/NES-MDCK reporter cell system for screening novel nuclear export inhibitors. NES signal-conjugated GFP accumulates in the nucleus in the presence of the nuclear export inhibitor leptomycin B (LMB). In this study, a stable GFP/NLS/NES fusion protein-expressing cell line was established, and its potential as a reporter was evaluated. The GFP/NLS/NES-MDCK cell line demonstrates improved nuclear accumulation of GFP in a time-course treatment with LMB. In addition, the dose-response data demonstrated superior sensitivity of GFP/NLS/NES-MDCK over GFP/NES-MDCK cells. As low as 0.01 ng/mL LMB is sufficient to cause accumulation of the GFP fusion protein in the nucleus in GFP/NLS/NES-MDCK cells, while at least 1 ng/mL of LMB is needed for the accumulation of GFP fusion protein in the nucleus of GFP/NES-MDCK cells. These results indicate that the newly established GFP/NLS/NES-MDCK cell line is a potentially powerful tool to screen for novel nuclear export inhibitors.
In order to develop an effective strategy of breast cancer therapy targeting survivin and its splice variants survivin-ΔEx3 and survivin-2B, the present study constructed four expression vectors by fusing the survivin antisense gene, the survivin (T34A) gene, the survivin-ΔEx3 antisense gene, and the survivin-2B gene with the enhanced green fluorescent protein (eGFP) gene. Each of these vectors was transiently transfected into the B-Cap-37 human breast cancer cell line. The effects of these four vectors with diverse genes on the proliferation and apoptosis of B-Cap-37 breast cancer cells were examined and compared in vitro using MTT and flow cytometry assays. Results of the MTT assay indicated that all four gene therapy plasmids were most effective at inhibiting the proliferation of B-Cap-37 cells 72 h after transfection. However, the four gene therapies had different rates of cell inhibition. pcDNA3.1(+)-egfp-anti-survivin and pcDNA3.1(+)-survivin (T34A)-egfp had almost equivalent or better effectiveness at suppressing cell growth. pcDNA3.1(+)-egfp-anti-survivin-ΔEx3 moderately inhibited the growth of B-Cap-37 cells. In contrast, pcDNA3.1(+)-survivin-2B-egfp had limited inhibition of cell growth. Similar profile of effectiveness of four gene therapies in soliciting cell apoptosis was also observed. These results suggest the relative importance of targeting survivin and its splice variant survivin-ΔEx3 in breast cancer treatment.
This study investigated the effect of ferulic acid (FA) on the up-regulation of heme oxygenase-1 (HO-1) in lymphocytes and the molecular mechanisms involved. Lymphocytes were treated with FA (0.001-0.1 μM) for certain times. Cell viability, the activity and level of expression of HO-1, and signal pathways were analyzed. FA significantly up-regulated HO-1 expression both at the level of mRNA and protein in lymphocytes. Moreover, FA-induced NF-E2-related factor (Nrf2) nuclear translocation and transcriptional activity, which is upstream of FAinduced HO-1 expression. In addition, lymphocytes treated with FA exhibited activation of extracellular regulated kinase (ERK) and treatments with U0126 (an ERK kinase inhibitor) attenuated the FA-induced activation of Nrf2, resulting in a decrease in HO-1 expression. Zinc protoporphyrin (ZnPP, a HO-1 inhibitor) markedly suppressed cytoprotection from radiation-induced cell damage by FA. Results suggested that the ERK signaling pathway controlled the anti-oxidation of FA by regulating the expression of the antioxidant enzyme HO-1.
As typical periodontopathic bacteria, Porphyromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) were exposed to electrolyzed ion-reduced water (ERI) and ERI containing 1% sodium carboxymethylcellulose (CMC-Na) (ERI-1% CMC-Na), and the time course of their bactericidal action was evaluated. More than 99% of each bacteria species were killed after exposure to each solution for 15 sec. In addition, 1% CMC-Na, which was added to prolong bactericidal action, did not affect the bactericidal action of ERI. Its bactericidal action was concentration-dependent. No viable P. gingivalis bacteria were observed at a concentration of 15% of the undiluted solution and no viable A. actinomycetemcomitans bacteria were observed at a concentration of 50%, indicating differences in the bactericidal action of ERI for the two bacteria species. These results suggest that ERI may be extremely useful in preventing and treating periodontal diseases.
Fenoterol HBr is a bronchodilator known to be subject to first pass effect after oral administration. The aim of this study was to prepare and evaluate fenoterol HBr suppositories. Suppositories were prepared by a fusion method using different fatty bases, viz. Witepsol H15, Witepsol E75, Suppocire AP, and Suppocire BM, as well as different hydrophilic bases, viz. polyethylene glycol and poloxamer bases. In vitro release studies revealed a greater release of the drug from hydrophilic bases than from fatty bases. The effect of incorporating different types and concentrations of non-ionic surfactants (Tween 60 and Span 20) on the release rate of the drug from Witepsol H15, as a model fatty base, was investigated. Results showed an enhanced release at low surfactant concentrations. A very fast 100% drug release was achieved when the drug was incorporated as an aqueous solution in Witepsol H15 (F17). This formula was selected to test the effect of fenoterol HBr suppositories on histamine-induced bronchospasms in Guinea pigs. No dyspnea of the animals was recorded for up to 30 min. In addition, thermogel liquid suppositories of different poloxamer 188 and poloxamer 407 proportions in the presence of sodium alginate as a mucoadhesive polymer were prepared. The different formulations behaved similarly concerning sustainment of drug release, however, only the formula containing 15% poloxamer 188 and 25% poloxamer 407 (F20) showed optimal gelation at body temperature. In conclusion, among the studied suppository bases there are bases suitable for fast release of the drug like F17 and hydrophilic bases especially polyethylene glycol, as well as other bases for sustained release applications of fenoterol HBr like fatty and thermogel bases.