Spinal cord injury (SCI) is a serious injury of the central nervous system and up until now there is no evident effective treatment for SCI. Axonal regeneration is the only way to restore functions after serious SCI that interrupt the long tracts mediating motor and sensory function. The hurdles for axonal regeneration in SCI include: glial scar tissue and molecular barriers, the inhibiting microenvironment, and the lack of sufficient neurotrophic support. Therefore, the key point of applying stem cells to treat SCI is to build a microenvironment conducive to the survival and differentiation of stem cells and regulate neurotrophic factor expression. Adult mesenchymal stem cells (MSCs) have been applied in experimental animal models and clinical trials of SCI. Genetic modification of MSCs can increase secretion of peptides or total length proteins with potential to repair SCI and promote survival of themselves and survival or regeneration of neurons. There are many proteins that have been applied to modified MSCs, such as neurotrophic factors (neurotrophin 3, brain-derived neurotrophic factor, glial cell line-derived neurotrophic factor, nerve growth factor, and MNTS1), receptor tyrosine kinases (tropomyosin-related kinase C), and hepatocyte growth factor. In the future, there will be more molecules acting as transgenes in MSCs for treatment of SCI.
The plasma membrane presents a remarkable barrier for the delivery of peptide and nucleic acid based drugs to the inside of cells. This restraint in the path of their development as therapeutic agents can be offset by their conjugation to cell penetrating peptides (CPPs) that can lead to an improved pharmacological profile. In this context, conformational behavior of Vimentin Tubulin Binding Site (TBS) peptide, Vim-TBS (58-81), was investigated for its acknowledged cell penetrating properties along with Trans-activating Tat (48-60) peptide and a pro-apoptogenic peptide of p21/WAFI protein (p10). Also, the fusion peptides Vim-TBS (58-81)-p10 & Tat (48-60)-p10 were studied using molecular mechanics (MM) and molecular dynamics (MD) based strategies. MM results revealed formation of stable α-helix like secondary structures in Vim-TBS (58-81), Tat (48-60) and p10 peptides. In water, three peptides adopted either a helical structure or a random conformation; the stability of either of the two states being governed by the formation of polar contacts with the solvent. The fusion peptides formed helical structures after MD simulations but the structure obtained for the fusion peptide, Vim-TBS-p10 is relatively better characterized in terms of its amphipathic nature with a hydrophilic face formed by the positively charged residues facilitating a better interaction of this fusion peptide with the membrane as compared to that of Tat-p10 peptide. This is the first report on the conformational characteristics of the Vim-TBS (58-81) peptide and the fusion peptide, Vim-TBS (58-81)-p10. The results presented here are significant for their potential role in guiding and facilitating the future efforts of designing peptide based cell penetrating drugs.
MLS128 monoclonal antibody, which binds an epitope consisting of two or three consecutive Tn-antigens, inhibits colon cancer cell growth by binding to a 110 kDa glycoprotein (GP). Previous studies suggested a possible association of insulin-like growth factor-I receptor (IGF-IR) signaling in the inhibition of colon cancer cell growth by MLS128 (Morita et al. Biosci Trends. 3, 32-37, 2009; Zamri et al. ibid. 6, 303-312, 2012). The current study thus investigated the nature of 110 kDa GP and its possible association with IGF-IR. MLS128 treatment for 3 days caused down-regulation of IGF-IR and disappearance of 110 kDa GP in HT29 colon cancer cells. Immunoprecipitation/immunoblotting experiments did not reveal a direct association between the two molecules in HT29 cells. In LS180 and HT29 cells, however, 110 kDa GP and IGF-IR were found in microdomains. Treatment of these cells with MLS128 for 3 days caused a reduction in the IGF-IR and 110 kDa GP associated with microdomains. Two-dimensional gel electrophoresis/MLS128 immunoblotting of HT29 and LS180 cell lysates and immunoprecipitates revealed three spots, from which tryptic peptides were recovered for protein sequencing. Identification of 110 kDa GP was unsuccessful due to its heterogeneity and resistance to tryptic digestion. During this study, however, limited proteolysis of 110 kDa GP was observed in the microdomain-associated 110 kDa GP from HT29 and LS180 cells, suggesting that protease-susceptible sites or domains exist in the middle of 110 kDa GP. This information on limited proteolysis may provide a clue to identifying 110 kDa GP.
The present study is designed to observe the inhibitory effect of compound A5, a semi- synthetic analogue of the natural compound andrographolide, on angiogenesis and its underlying mechanism. Compound A5 is semi-synthesized from natural compound neoandrographolide. Andrographolide, the aglycon of neoandrograoholide, and A5 all inhibited vascular endothelial growth factor (VEGF)-induced human umbilical vein endothelial cells (HUVECs) proliferation, and that the inhibition shown by A5 is the best. A5 also inhibited VEGF-induced tube formation in HUVECs in a concentration-dependent manner. VEGF-induced neoangiogenesis in vivo was observed by Matrigel formation assay. The Matrigel picture and CD31 staining results showed that A5 inhibited VEGF-induced neoangiogenesis in vivo. Further, Western-blot results showed that A5 inhibited VEGF- induced phosphorylation of VEGF receptor 2 (VEGFR2), extracellular signal-regulated kinase 1 and 2 (ERK1/2), and p38 kinase. The antitumor effect of A5 was analyzed in a xenograft mouse tumor model inoculated with hepatoma Hep3B cells. The results showed that A5 decreased tumor weight and tumor size without affecting body weight in the xenograft mouse, and A5 also decreased CD31 staining in tumor tissue. Taken together, the present study demonstrates that compound A5 inhibits tumor growth via blocking neoangiogenesis, and the cellular VEGFR2-p38/ERK1/2 signal pathway.
SL1122-37 is a novel derivative of sorafenib that was characterized by introducing trifluoromethyl on the 4-position of indazole. We aimed to evaluate the effects of SL1122-37 on human hepatocellular carcinoma (HCC) growth and on umbilical vein vascular endothelial cells (HUVECs) angiogenesis. Its efficacy and mechanisms were compared with sorafenib. SL1122-37 significantly prevented PLC/PRF/5 cell proliferation as estimated by colorimetric assay. Flow cytometry analysis showed the induction of apoptosis and arrest of cell cycle in G1 phase. Western blotting showed the decrease of cyclin D1 and regulation of apoptotic proteins. Further analysis suggested that these effects of SL1122-37 might arise from its roles in the inhibition of multi-kinases, including c-Kit and its downstream targets and the Wnt/β-catenin pathway in PLC/PRF/5 cells. SL1122-37 also possessed the activity of antiangiogenesis, showing the prevention of HUVEC migration and capillary tube formation. Western blotting indicated the inhibition of VEGF and phosphorylation of VEGFR-2 in HUVECs. Statistical analysis suggested that SL1122-37 might possess greater activities than sorafenib in the prevention of HCC proliferation and HUVEC angiogenesis. Conclusion, SL1122-37 could develop as a potent anticancer agent for the treatment of HCC.
Like health care systems in other developed countries, Japan’s health care system faces significant challenges due to aging of the population and economic stagnation. Advanced medicine (Senshin Iryou) is a unique system of medical care in Japan offering highly technology-driven medical care that is not covered by public health insurance. Advanced medicine has recently developed and expanded as part of health care reform. Will it work? To answer this question, we briefly trace the historical development of advanced medicine and describe the characteristics and current state of advanced medical care in Japan. We then offer our opinions on the future of advanced medicine with careful consideration of its pros and cons. We believe that developing advanced medicine is an attempt to bring health care reform in line rather than the goal of health care reform.
Evidence-based clinical practice guidelines (CPGs) have been used in many countries around the world to promote standardized management of hepatocellular carcinoma (HCC). Guidelines implemented in Japan provide a good example of "translating the best current research evidence into clinical practice and obtaining new evidence in the course of influencing practitioners' attitudes and clinical decision-making". The Japanese guidelines have achieved remarkable results in terms of HCC management in Japan. The first Japanese evidence-based CPGs for HCC (J-HCC Guidelines) were published in 2005 and then revised in 2009. A second updated version that incorporates new evidence was just published on October 15, 2013. China accounts for 55% of HCC cases worldwide. Although the Government devised a series of directives on the management of HCC and Chinese Guidelines on HCC were published in 2009, neither were based on systematic review and evaluation of the literature and neither included recommendations supported by data. The novel concept of "precision surgery" was recently proposed in China. This concept is based on surgeons' clinical experience and should encourage the standardized management of HCC in China. However, recommendations supported by data are still urgently needed to guide clinical decision-making in order to facilitate standardized management of HCC in China.
Influenza pandemics are a serious threat to public health in today's world. In the past 10 years, the outbreak of three forms of severe influenza – H5N1, H1N1, and H7N9 – has caused tremendous loss of life and property. In order to better cope with pandemics, antivirals such as oseltamivir are being stockpiled in great quantities, placing a substantial burden on government budgets and potentially resulting in massive waste because of the uncertainty as to when an influenza pandemic will strike and whether emerging virus strains will be resistant to the stockpiled drugs. Complementary and alternative medicine (CAM) is generally available, affordable, and commonly used in China and many other countries and CAM has a long track record of fighting influenza. The Chinese Government appropriated funds to intensively investigate herbal medicines in accordance with the principles of evidence-based medicine in order to identify effective, inexpensive, and easily stockpiled medicines. Thus far, several drugs including Shufeng Jiedu capsules, Lianhua Qingwen capsules, Maxing Shigan decoction, Yinqiao powder, and Jinhua Qinggan granules have demonstrated effectiveness in fighting influenza. In the future, CAM is expected to make greater contribution in controlling the prevalence of influenza pandemics.
In the spring of 2013, an emerging infectious disease emerged in China, 132 cases of human were infected with the H7N9 avian influenza virus, 39 cases were resulted in death within 3 month, which sparked a global concern about public health. Many reports have been published about this disease, including clinical characteristics and genomic information. However, more emerging infectious disease may infect human in the future. Confronted with the escalating scale of compounding probabilities, physicians or clinicians as the first line that meet patients who suffering from emerging infectious disease, we should do better by using our intellect and strong will to carry out public health measures, biomedical research, and technological advances.