Comparison of liposome-polycation-DNA(LPD) and monophosphoryl lipid A(MPL) adjuvant formulations in BALB/c mice models.

It is of fundamental importance to use an appropriate adjuvant to generate a potent immune response for immunotherapy. In this study, we had a comparative investigation on the effectiveness of two adjuvant formulations, liposome-polycation-DNA (LPD) and monophosphoryl lipid A(MPL) in combination with a truncated peptide of bFGF(tbFGF) as antigen. LPD/tbFGF induced continuously increasing antibodies expression during the whole immunization period. In contrast, the level of antibodies was variable in MPL/tbFGF-immunized mice, MPL/tbFGF elicited potent antibodies response in the early-phase of immunization (during the first 3 immunizations), but the later immunizations did not produce a significant increase in the level of antibodies. Evaluation of IFN-γ and IL-4 responses revealed that both LPD/tbFGF and MPL/tbFGF demonstrated generation of higher level of IFN-γ, whereas no significant increase in IL-4 levels was detected in the two groups. In addition, histological analysis exhibited obvious germinal centers in the spleen tissues of LPD/tbFGF mice. The data suggested that LPD would be a promising long-effective adjuvant due to its potent and persistent immunostimulation and MPL could play an appropriate role in short-acting immunization.

[Complications and the management of fully covered retrievable metal stent placement in benign tracheal stenosis].

OBJECTIVE: To study complications and the management of the use of covered retrievable expandable metallic stents in the treatment of benign tracheal stenosis. METHODS: Fully covered retrievable metal stents were placed in 21 patients with benign tracheal stenosis. Stent-related complications and the management were reviewed and analysed. RESULTS: Twenty-eight fully covered retrievable metallic stents were successfully placed fluoroscopically in 21 patients. Stents were removed 4 - 12 months [mean (5.5 ± 2.2) mouths] after placement in all patients. Stent-related complications included granulation tissue (n = 18), stent migration (n = 4), stent expectoration (n = 2), halitosis (n = 8), mucous retention (n = 21) and mucus plugging (n = 1). Granulation tissue was removed with a carbon dioxide laser in 2 patients. Stents were replaced for 2 times and 3 times respectively in 2 patients after stent migration and stent expectoration. An additional stent was placed in 2 patients after stent migration. Symptom of halitosis was relieved after prolonged course of systemic antibiotics treatment in 8 patients. Symptom of mucous retention was relieved with nebulized saline and N-acetylcysteine saline inhalation. Mucous plug was expelled after severe coughing after suctioning using an aspirator in one patient. There were statistically significant differences in stricture diameter, rank of tachypnea and pulmonary function (FEV(1)) in all 21 patients before stent insertion and after stents removal. No patient has experienced recurrence during the follow-up period of 1 - 36 months [mean (23.2 ± 8.0) months]. CONCLUSION: Fully covered retrievable metallic stent may be a safe and effective in benign tracheal stenosis. Stent-related complications may be effectively managed.

Europium-Pyridinedicarboxylate-Adenine Light-Up Fluorescence Nanoprobes for Selective Detection of Phosphate in Biological Fluids.

Phosphate (Pi) plays important roles in various physiological processes. Its quantification in biological fluids is highly crucial for timely warning of Pi accumulation. Herein, an europium (Eu)-based coordination polymer nanoprobe (Eu/DPA/Ade) is prepared by coordinating 2,6-pyridinedicarboxylic acid (2,6-DPA) and adenine (Ade) with Eu3+. Eu/DPA/Ade exhibits light-up fluorescence response to Pi. The strong coordinating interaction between Eu3+ and O atoms in the Pi group not only shortens the Eu3+-ligand distance to improve the energy transfer from 2,6-DPA to Eu3+ but also attenuates the fluorescence quenching from water molecules in the coordinating sphere of Eu3+. Eu/DPA/Ade produces red emission at λem 618 nm via the "antenna effect". The coligand Ade further promotes the fluorescent emission. The selective recognition of Pi within 10-60 µM is achieved with a detection limit of 4.65 µM. In addition, a certain level of Pi (100-170 µM) causes an exponential increment on the fluorescence of Eu/DPA/Ade and makes it feasible for visual estimation of Pi under irradiation by an ultraviolet lamp at 254 nm. The quantitative detection and visual estimation of Pi in human urine and saliva have been demonstrated.

Astragalus Saponins and Liposome Constitute an Efficacious Adjuvant Formulation for Cancer Vaccines.

Cancer vaccines mostly aim to induce cytotoxic T lymphocytes (CTLs) against tumors. An appropriate adjuvant is of fundamental importance for inducing cellular immune response. Since the antigen in particulate form is substantially more immunogenic than soluble form antigen, it is beneficial to interact with antigen-presenting cells membrane to induce robust CD8+ T cell activation following vaccination. Based on previous research, we designed an adjuvant formulation by combining Astragalus saponins, cholesterol, and liposome to incorporate antigen into a particulate delivery system, so as to enhance cellular immune response. Meanwhile, angiogenesis contributes to tumor growth and metastasis, and basic fibroblast growth factor (bFGF) is involved in tumor angiogenesis. Therefore, using lipo-saponins adjuvant formulation and a human recombinant bFGF antigen protein, we tried to induce bFGF-specific CTL response to inhibit tumor angiogenesis to achieve antitumor activity. After five immunizations, the lipo-saponins/bFGF complex elicited robust antibody response and markedly higher amount of interferon-γ in BALB/c mice, resulting in superior antitumor activities. Decreased microvessel density in CD31 immunohistochemistry and the lysis of vascular endothelial cells by the T lymphocytes from the immunized mice indicated that the immunity inhibited the angiogenesis of tumors and further led to the inhibition of tumors. Our data suggest that the approach to construct adjuvant formulation between liposome and Astragalus saponins appeared highly desirable, and that Astragalus saponins may be utilized as a valuable additive for enhancing the effectiveness of vaccines and stimulating an appropriate immune response that can benefit tumor therapy.

[Influence of different restorations on the fracture resistance of endodontically treated premolars with minor loss of dental defect].

PURPOSE: To evaluate the influence of 3 different restorations on the fracture resistance of endodontically treated premolars with minor loss of dental defect in order to direct clinical design. METHODS: Seventy-two recently extracted intact and single-rooted human mandibular premolars for orthodontic reason were collected and randomly divided into 3 groups (group A, B, C, n=24). Each group was respectively prepared into 1/3 defect of occlusal surface (group A), 1/2 defect of occlusal surface (group B) and 1/3 defect of proximal-occlusal surface model (group C). Then each group was divided into 3 subgroups with one restored with a light-curing composite resin(group A1, B1, C1), one restored with cast metal full crowns following a light-curing composite resin (group A2, B2, C2), and one restored with fiber posts and resin cores and cast full crowns after teeth preparations (group A3, B3, C3, n=8). Static loading tests were performed on each specimen until cracked.Fracture strength was tested and fracture patterns were examined. The data was analyzed using SPSS17.0 software package. RESULTS: The fracture resistance of subgroup A2 was different when compared with subgroup A1 and subgroup A3. In group B and C, the fracture resistance of teeth in subgroup B2, B3, C2 and C3 were significantly higher than that in subgroup B1 and C1. There was no significant difference in the fracture mode of 3 restorations in group A, B and C. CONCLUSIONS: A composite resin combined with cast metal full crowns can be used as the first choice to restore endodontically treated premolars with proximal-occlusal 1/3 defects and occlusal defects that do not surpass 1/2 dimension of occlusion.

[Effects of Different Residue Part Inputs of Corn Straws on CO2 Efflux and Microbial Biomass in Clay Loam and Sandy Loam Black Soils].

The decomposed rate of crop residues is a major determinant for carbon balance and nutrient cycling in agroecosystem. In this study, a constant temperature incubation study was conducted to evaluate CO2 emission and microbial biomass based on four different parts of corn straw (roots, lower stem, upper stem and leaves) and two soils with different textures (sandy loam and clay loam) from the black soil region. The relationships between soil CO2 emission, microbial biomass and the ratio of carbon (C) to nitrogen (N) and lignin of corn residues were analyzed by the linear regression. Results showed that the production of CO2 was increased with the addition of different parts of corn straw to soil, with the value of priming effect (PE) ranged from 215. 53 µmol . g-1 to 335. 17 µmol . g -1. Except for corn leaves, the cumulative CO2 production and PE of clay loam soil were significantly higher than those in sandy loam soil. The correlation of PE with lignin/N was obviously more significant than that with lignin concentration, nitrogen concentration and C/N of corn residue. The addition of corn straw to soil increased the contents of MBC and MBN and decreased MBC/MBN, which suggested that more nitrogen rather than carbon was conserved in microbial community. The augmenter of microbial biomass in sandy loam soil was greater than that in clay loam soil, but the total dissolved nitrogen was lower. Our results indicated that the differences in CO2 emission with the addition of residues to soils were primarily ascribe to the different lignin/N ratio in different corn parts; and the corn residues added into the sandy loam soil could enhance carbon sequestration, microbial biomass and nitrogen holding ability relative to clay loam soil.

Acupuncture combined with curcumin disrupts platelet-derived growth factor ß receptor/extracellular signal-regulated kinase signalling and stimulates extracellular matrix degradation in carbon tetrachloride-induced hepatic fibrosis in rats.

BACKGROUND: Acupuncture treatment has been increasingly used to treat chronic liver diseases. We previously reported that acupuncture combined with curcumin, a natural antifibrotic compound, could remarkably attenuate liver fibrosis in chemically intoxicated rats, but the underlying molecular mechanisms are poorly understood. The present study was aimed at investigating the effects of acupuncture combined with curcumin on platelet-derived growth factor (PDGF) signalling and extracellular matrix (ECM) regulation in the fibrotic liver. METHODS: A total of 60 Sprague-Dawley male rats were randomly divided into control, model, sham, acupuncture, curcumin and combination treatment groups. During the establishment of fibrosis using carbon tetrachloride (CCl(4)), acupuncture at LR3, LR14, BL18 and ST36 and/or curcumin treatment by mouth were performed simultaneously. After treatment, serum PDGF levels were measured. Protein and mRNA expression of key effectors in PDGF pathway and fibrinolysis in the liver was determined. RESULTS: Acupuncture combined with curcumin potently reduced serum PDGF levels and selectively disrupted the PDGF-ßR/extracellular signal-regulated kinase (ERK) cascade. Combination treatment also significantly repressed expression of connective tissue growth factor and upregulated expression of matrix metalloproteinase-9, promoting fibrinolysis in the fibrotic liver. CONCLUSIONS: The beneficial effects of acupuncture and its combination with curcumin could be attributed to the disruption of PDGF-ßR/ERK pathway and stimulated ECM degradation in the fibrotic liver. Acupuncture treatment significantly enhanced curcumin effects at the molecular level. These findings may provide molecular insights into the potential of acupuncture combined with curcumin for prevention of hepatic fibrosis.

Acupuncture combined with curcumin attenuates carbon tetrachloride-induced hepatic fibrosis in rats.

BACKGROUND: Increasingly, studies demonstrate the effectiveness of acupuncture therapy against liver fibrosis. Curcumin is a natural product with antifibrotic effects, but has poor pharmacokinetic profiles. This study aimed to evaluate whether acupuncture combined with curcumin could more potently attenuate liver fibrosis in chemical intoxicated rats. METHODS: 60 Sprague-Dawley male rats were randomly divided into control, model, sham, acupuncture, curcumin and combination therapy groups. During the establishment of fibrosis using carbon tetrachloride (CCl(4)), acupuncture at LR3, LR14, BL18 and ST36 and/or curcumin treatment by mouth were performed simultaneously. After treatment, pathological indexes and histology for hepatic injury and fibrogenesis were detected. The expression of extracellular matrix (ECM) components was also determined. RESULTS: Acupuncture combined with curcumin potently protected the liver from CCl(4)-induced injury and fibrogenesis, as indicated by reduced levels of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, hyaluronic acid, laminin and procollagen III. Combined use also led to significant liver histological improvements. Furthermore, combined use effectively inhibited ECM expression such as α-smooth muscle actin, fibronectin and α1(1) collagen. CONCLUSIONS: Acupuncture treatment could significantly enhance the antifibrotic efficacy of curcumin on CCl(4)-induced hepatic fibrosis in rats in vivo, suggesting that a combination of acupuncture with curcumin may be exploited for the prevention of hepatic fibrosis.

An N-, C-terminally truncated basic fibroblast growth factor and LPD (liposome-polycation-DNA) complexes elicits a protective immune response against murine colon carcinoma.

Basic fibroblast growth factor (bFGF) is a mitogen for endothelial cells, which participates in tumor angiogenesis. Active immunity against bFGF could be a promising approach for the biotherapy of cancer. Because bFGF is abundant in normal and malignant tissues, it is presumably difficult for normal bFGF to induce immunity due to self-tolerance. In addition, previous studies have shown that a complex consisting of a cationic liposome and a non-coding plasmid DNA can be used to stimulate innate immunity. This stimulation initiates a potent cytokine response, which can inhibit tumor growth. To investigate the effects of immunity against bFGF on murine colon carcinomas, we employed an N-, C-terminally truncated basic fibroblast growth factor (tbFGF, of human origin) as an antigen and a liposome-DNA complex as an adjuvant. After six immunizations, a robust bFGF-specific immune response was elicited. Subsequently, inhibition of tumor growth and a significant reduction in tumor vasculature were observed. The antitumor effect was confirmed by adoptive therapy of activated spleen cells from the immunized mice. In vitro, a CTL assay revealed that bFGF-specific cytotoxic T lymphocytes (CTL) resulted in the lysis of mouse microvascular endothelial cells (MS1) rather than that of the CT26 colorectal cancer cells. These results suggest that anti-angiogenesis treatment induced by a bFGF-specific CTLs against microvascular endothelial cells may be a useful method for cancer therapy.