Developing Chinese herbal-based functional biomaterials for tissue engineering.

The role of traditional Chinese medicine (TCM) in treating diseases is receiving increasing attention. Chinese herbal medicine is an important part of TCM with various applications and the active ingredients extracted from Chinese herbal medicines have physiological and pathological effects. Tissue engineering combines cell biology and materials science to construct tissues or organs in vitro or in vivo. TCM has been proposed by the World Health Organization as an effective treatment modality. In recent years, the potential use of TCM in tissue engineering has been demonstrated. In this review, the classification and efficacy of TCM active ingredients and delivery systems are discussed based on the TCM theory. We also summarized the current application status and broad prospects of Chinese herbal active ingredients in different specialized biomaterials in the field of tissue engineering. This review provides novel insights into the integration of TCM and modern Western medicine through the application of Chinese medicine in tissue engineering and regenerative medicine.

Non-viral Gene Therapy for Melanoma Using Lysenin from Eisenia Foetida.

Earthworms, long utilized in traditional medicine, serve as a source of inspiration for modern therapeutics. Lysenin, a defensive factor in the coelom fluid of the earthworm Eisenia fetida, has multiple bioactivities. However, the inherent toxicity of Lysenin as a pore-forming protein (PFP) restricts its application in therapy. Here, a gene therapy strategy based on Lysenin for cancer treatment is presented. The formulation consists of polymeric nanoparticles complexed with the plasmid encoding Lysenin. After transfection in vitro, melanoma cells can express Lysenin, resulting in necrosis, autophagy, and immunogenic cell death. The secretory signal peptide alters the intracellular distribution of the expressed product of Lysenin, thereby potentiating its anticancer efficacy. The intratumor injection of Lysenin gene formulation can efficiently kill the transfected melanoma cells and activate the antitumor immune response. Notably, no obvious systemic toxicity is observed during the treatment. Non-viral gene therapy based on Lysenin derived from Eisenia foetida exhibits potential in cancer therapy, which can inspire future cancer therapeutics.

Intraspecific Identification of Lingzhi Medicinal Mushroom, Ganoderma lingzhi (Agaricomycetes), Using ISSR Markers.

This study aimed to obtain a set of specific inter simple sequence repeat (ISSR) primers and establish a stable and accurate intraspecific identification method for Ganoderma lingzhi. A total of 117 G. lingzhi strains were identified using internal transcribed spacer sequences from 147 strains determined as G. lingzhi via simple morphological identification. Based on the sequences obtained, specific ISSR primers for G. lingzhi were screened and validated, and 15 specific ISSR primers showed polymorphic banding pattern with clear band resolution. Subsequently, ISSR PCRs of the 15 specific primers were performed for the 117 G. lingzhi strains. As expected, DNA analysis of the ISSR markers could distinguish G. lingzhi strains, with similarity coefficients ranging from 0.11 to 0.89. Thus, the 15 specific ISSR primers can be used for intraspecific identification and polymorphism analysis of G. lingzhi.

Dehydropachymic acid decreases bafilomycin A1 induced ß-Amyloid accumulation in PC12 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Fuling, the sclerotium of Poria cocos, was frequently used in traditional Chinese medicine (TCM) formulae for Alzheimer's disease (AD) intervention over the past 10 centuries. And its extracts exhibited significant effects in both cellular and animal models of AD in previous studies. However, its mechanisms on prevention and treatment of AD have not been well elucidated yet. AIM OF THE STUDY: To investigate the effect and corresponding mechanisms of dehydropachymic acid, which is one of the major triterpenes in P. cocos, on the clearance of ß-amyloid accumulation in bafilomycin A1 induced PC12 cells. MATERIALS AND METHODS: MTT assay was used to examine the DPA effect on the viability of PC12 cells stable transfected with pCB6-APP (PC12-APP). PC12-APP cells were treated with DPA at the concentration of 6.25, 12.5, 25µg/mL for 4h, and then co-treated with 50nmol/L bafilomycin A1 for 48h except the controls. The Aß1-42 content in culture medium was determined by ELISA. The intracellular amount of APP, Aß1-42, LC3, cathepsin D was measured by Western blotting and normalized to GAPDH loading control. The PC12 cells stable transfected with pSelect-LC3-GFP (PC12-LC3-GFP) was used in the fluorescence microscopy estimation of autophagosomes accumulation. The internal pH in lysosome was detected by LysoTracker Red staining. RESULTS: DPA had no significant effect on the cell viability but could significantly decrease Aß1-42 content in culture medium and eliminate the intracellular accumulation of APP and Aß1-42 in bafilomycin A1 induced PC12-APP cells. Furthermore, DPA lowered the LC3-II/LC3-I ratio and reduced the GFP-labeled LC3 puncta which were elevated by bafilomycin A1. And the increase in internal pH of lysosome and decrease in mCatD amount in Bafilomycin A1 induced PC12-APP cells were restored by DPA treatment. These results indicated that DPA could restore the lysosomal acidification and recover the autophgic flux which is impaired by bafilomycin A1. CONCLUSIONS: DPA could effectively clear the accumulation of Aß1-42 in bafilomycin A1 impaired PC12 cells through restoring the lysosomal acidification and recovering the autophgic flux. And these results highlight its therapeutic potential for AD treatment.

Metagenomic Analysis of Genes Encoding Nutrient Cycling Pathways in the Microbiota of Deep-Sea and Shallow-Water Sponges.

Sponges host complex symbiotic communities, but to date, the whole picture of the metabolic potential of sponge microbiota remains unclear, particularly the difference between the shallow-water and deep-sea sponge holobionts. In this study, two completely different sponges, shallow-water sponge Theonella swinhoei from the South China Sea and deep-sea sponge Neamphius huxleyi from the Indian Ocean, were selected to compare their whole symbiotic communities and metabolic potential, particularly in element transformation. Phylogenetically diverse bacteria, archaea, fungi, and algae were detected in both shallow-water sponge T. swinhoei and deep-sea sponge N. huxleyi, and different microbial community structures were indicated between these two sponges. Metagenome-based gene abundance analysis indicated that, though the two sponge microbiota have similar core functions, they showed different potential strategies in detailed metabolic processes, e.g., in the transformation and utilization of carbon, nitrogen, phosphorus, and sulfur by corresponding microbial symbionts. This study provides insight into the putative metabolic potentials of the microbiota associated with the shallow-water and deep-sea sponges at the whole community level, extending our knowledge of the sponge microbiota's functions, the association of sponge- microbes, as well as the adaption of sponge microbiota to the marine environment.

[Progress on the research of acupuncture for smoking cessation in foreign and domestic].

Up to December of 2011, the domestic and overseas literatures of acupuncture for smoking cessation are searched and collected through Pubmed, CNKI, Wanfang and Chongqing VIP databases, which are analyzed from treatment method, action mechanism, influencing factors of efficacy and efficacy evaluation research and so on. The result shows that (1) acupuncture has positive effect on smoking cessation, and large clinical trials has been carried out. However, most of them are needling methods and the short-term effect observation, which lack of long-term efficacy research of withdrawal symptoms, especially continuous withdrawal symptoms and index of life quality. (2) Because of unified clinical treatment, manipulation standard and curative effect criteria, it is difficult to make comparison among them. (3) The study of mechanism on acupuncture for smoking cessation is not of in-depth. In future, clinical research project of acupuncture on smoking cessation should be optimized and regulated, including the unified treatment, manipulation standard and curative effect criteria, also long-term researches with large-scale samples should be launched to confirm effect ofacupuncture for smoking cessation to further discuss its action mechanism.

[The invasion ability and expressions of matrix metalloproteinase-13 and calcium-binding protein S100A4 are inhibited by hyperthermia in human Tca8113 cells].

OBJECTIVE: To study the effect of hyperthermia on anti-invasion of Tca8113 and the expression change of matrix metalloproteinase-13 (MMP-13) and calcium-binding protein S100A4 (S100A4). METHODS: Tca8113 cell pools were incubated at 43 degrees C for 0, 40, 80, 120 min, respectively, and at 37, 41, 43, 45 degrees C respectively for 80 min. The effect of high temperatures on the invasion ability of Tca8113 was measured in vitro. The slides of cells were made and incubated at 43 degrees C for 0, 40, 80, 120 min, respectively. Immunocytochemical method was employed for detecting the expression change of MMP-13 and S100A4 protein. Tca8113 cells were incubated at 43 degrees C for 0, 40, 80, 120 min respectively and at 37, 41, 43, 45 degrees C respectively for 80 min. Western blot method was conducted for detecting the expressionchange of MMP-13 and S100A4 protein. RESULTS: As incubating time at higher temperature lasted, the proportion of the cells with invasion ability decreased. Except groups of 40 min and 80 min at 43 degrees C and 41, 43 degrees C for 80 min, the rest groups show significant statistic differences (P < 0.05). The expression intensity of MMP-13 and S100A4 proteins in Tca8113 cells would decrease as incubating time at higher temperature lasted. The content of MMP-13 and S100A4 proteins would decrease as incubating time at higher temperature lasted or incubating temperature increased. Except the groups of 40, 80 min at 43 degrees C and 41, 43 degrees C for 80 min, statistic differences were identified (P < 0.05). CONCLUSION: The invasion of Tca8113 could be inhibited by hyperthermia. The mechanism of this effect may be due to protein expression inhibition of MMP-13 and S100A4.

Variability of fecal indicator bacteria in flowing and ponded waters in southern California: implications for bacterial TMDL development and implementation.

Recreational water quality is assessed by using water quality objectives for fecal indicator bacteria (FIB) including total coliform, fecal coliform (or E. coli), and/or Enterococcus. It is required under the Clean Water Act that a TMDL be developed for a bacteria-impaired water body. The development and implementation of bacterial TMDLs has proven challenging and often difficult due to unknown source(s) of FIB. This study found that FIB levels varied significantly in flowing water, ponded water, and associated sediment. FIB levels in isolated ponded water in waterways were significantly higher than in flowing water. Sediment under ponded water contained a great amount of FIB. Furthermore, FIB concentrations in ponded water tended to increase with increasing water temperature and to decrease with increasing water salinity. The result provides the field evidence of survival/growth of FIB in water and sediment under ambient conditions in southern California. A holistic approach including natural sources (e.g., a reference system) should be considered for practical and applicable purposes while developing and implementing bacterial TMDLs for pathogen-impaired waterbodies.

[Study on effect of berbamine on multidrug resistance leukemia K562/Adr cells].

OBJECTIVE: To study the effect and mechanism of berbamine on the apoptosis of multidrug resistant leukemia K562/Adr cells and in reversing the drug resistance. METHODS: IC50 value of K562/Adr cell was determined with MTT method, cell apoptosis rate was analyzed by flow cytometry with Annexin V FITC-PI assay, with the peak and cell cycle detected by PI staining. At the same time, flow cytometry was also used in determining Caspase-3, P-GP protein expression and drug accumulating capacity in cells, and RT-PCR method was used to analyze the gene expression of mdr-1. RESULTS: Berbamine could inhibit human leukemia K562/Adr cell growth in dose-dependent manner, it could also induce cell apoptosis, increase the protein expression of Caspase-3 and the drug excretion capacity of cells, reduce the mRNA and protein expression levels of mdr-1 gene. CONCLUSION: Berbamine could activate Caspase-3 to induce human leukemia K562/Adr cell apoptosis, and by reducing mdr-1 gene expression to reverse its multidrug resistance.