The pharmacokinetics of small interfering RNAs (siRNAs) is a pivotal issue for siRNA-based drug development.In this study, we comprehensively investigated the behavior of siRNAs in vivo in various tissues and demonstrated that intravenously-injected naked siRNA accumulated remarkably in the submandibular gland, bulbourethral gland, and pancreas, with a respective half-life of ~22.7, ~45.6, and ~30.3 h.This was further confirmed by gel separation of tissue homogenates and/or supernatants.In vivo imaging and cryosectioning suggested that delivery carriers significantly influence the distribution and elimination profiles of siRNA.Gene-silencing assays revealed that neither naked nor liposome-formulated siRNA resulted in gene knockdown in the submandibular and bulbourethral glands after systemic administration, suggesting that these glands function as drug reservoirs that enable slow siRNA release into the circulation.But robust gene-silencing was achieved by local injection of liposome-encapsulated siRNA into the submandibular gland.Our results enhance understanding of the pharmacokinetic properties of siRNAs and we believe that they will facilitate the development of siRNA therapy, especially for the submandibular gland.
Effects of potato-onion extraction concentration on Fusarium oxysporum spore germination,hyphae growth,fungi biomass,and spore production were evaluated in this research.Results showed that potato-onion extraction concentration had inhibition on Fusarium oxysporum spore germination rate,fungi biomass,and spore production to some extent.The hyphae growth inhibition reached the highest value with 69.21% with treated by 1 000 mg·mL-1 of potato-onion extraction.The aim of this research was to explore the biocontrol feasibility of potato-onion extraction concentration on Fusarium oxysporum and provide theory in practice control.
To determine the role of miR-25 in non-small cell lung cancer (NSCLC), we first detected miR-25 expression in clinical specimens and lung cancer cell lines by quantitative real-time polymerase chain reaction. The levels of miR-25 were elevated in the plasma of NSCLC patients and NSCLC cell lines. Transfection of A549 and 95-D cells with a miR-25 inhibitor resulted in reduced cell proliferation and enhanced apoptosis. Moreover, the modulator of apoptosis 1 (MOAP1) gene was identified as a novel target of miR-25. The ability of miR-25 to promote cell proliferation and block apoptosis is attributable to its effect on MOAP1 suppression. In addition, miR-25 antagomir significantly inhibited lung cancer growth via upregulation of MOAP1 in a mouse xenograft model. Collectively, these data demonstrate that miR-25 is an important biomarker for lung cancer, and miR-25 promotes cell proliferation and inhibits apoptosis in NSCLC cells by negatively regulating MOAP1 expression.
Hydrophobization of cationic polymers, as an efficient strategy, had been widely developed in the structure of cationic polymer micelles to improve the delivery efficiency of nucleic acids. However, the distribution of hydrophobic segments in the polymer chains is rarely considered. Here, we have elaborated three types of hydrophobized polyethylene glycol (PEG)-blocked cationic polymers with different distributions of the hydrophobic segments in the polymer chains PEG–PAM–PDP (E–A–D), PEG–PDP–PAM (E–D–A), and PEG–P(AM/DP) (E–(A/D)), which were synthesized by reversible addition–fragmentation chain transfer polymerization of methoxy PEG, cationic monomer aminoethyl methacrylate, and pH-sensitive hydrophobic monomer 2-diisopropylaminoethyl methacrylate, respectively. In aqueous solution, all of the three copolymers, E–A–D, E–D–A, and E–(A/D), were able to spontaneously form nanosized micelles (100–150 nm) (ME–A–D, ME–D–A, and ME–(A/D)) and well-incorporated small interfering RNA (siRNA) into complex micelles (CMs). The effect of distributions of the hydrophobic segments on siRNA delivery had been evaluated in vitro and in vivo. Compared with ME–D–A and ME–(A/D), ME–A–D showed the best siRNA binding capacity to form stable ME–A–D/siRNA CMs less than 100 nm, mediated the best gene-silencing efficiency and inhibition effect of tumor cell growth in vitro, and showed better liver gene-silencing effect in vivo. In the case of ME–(A/D) with a random distribution of cationic and hydrophobic segments, a gene-silencing efficiency higher than Lipo2000 but lesser than ME–A–D and ME–D–A was obtained. As the mole ratio of positive and negative charges increased, ME–D–A/siRNA and ME–A–D/siRNA showed similar performances in size, zeta potential, cell uptake, and gene silencing, but ME–(A/D)/siRNA showed reversed performances. In addition, ME–A–D as the best siRNA carrier was evaluated in the tumor tissue in the xenograft murine model and showed good anticancer capacity. Obviously, the distribution of the hydrophobic segments in the amphiphilic cationic polymer chains should be seriously considered in the design of siRNA vectors.
Millions of workers in China rely on respirators and other personal protective equipment to reduce the risk of injury and occupational diseases. However, it has been >25 years since the first survey of facial dimensions for Chinese adults was published, and it has never been completely updated. Thus, an anthropometric survey of Chinese civilian workers was conducted in 2006. A total of 3000 subjects (2026 males and 974 females) between the ages of 18 and 66 years old was measured using traditional techniques. Nineteen facial dimensions, height, weight, neck circumference, waist circumference and hip circumference were measured. A stratified sampling plan of three age strata and two gender strata was implemented. Linear regression analysis was used to evaluate the possible effects of gender, age, occupation and body size on facial dimensions. The regression coefficients for gender indicated that for all anthropometric dimensions, males had significantly larger measurements than females. As body mass index increased, dimensions measured increased significantly. Construction workers and miners had significantly smaller measurements than individuals employed in healthcare or manufacturing for a majority of dimensions. Five representative indexes of facial dimension (face length, face width, nose protrusion, bigonial breadth and nasal root breadth) were selected based on correlation and cluster analysis of all dimensions. Through comparison with the facial dimensions of American subjects, this study indicated that Chinese civilian workers have shorter face length, smaller nose protrusion, larger face width and longer lip length.
Objective To observe the effect of leptin on proliferation and apoptosis of breast cancer MCF-7 cell line,and to explore the effect of leptin on occurrence and development of breast cancer.Method The MCF-7 cell line was treated with different concentration of leptin in vitro.Cell proliferation was evaluated by MTT assay. Distribution of cell cycle was determined by flow cytomery, meanwhile the rates of apoptosis were estimated on the basis of Annexin V-FITC/PI apoptosis detection. Results When treated with different concentration of leptin for 24 h, 48 h and 72 h, they could significantly induce the proliferation of MCF-7 cells by MTT method.There was not interaction between concentration of leptin and time course (F=0.919,P=0.523).The main effect of concentration of leptin and time course was statistically significant (F=12.699,P=0.000;F=647.881, P=0.000). Compared 200 ng/ml and 400 ng/ml with the control group, we found the difference was statistically significant by multiple comparison (P=0.007,P=0.000,respectively).The difference was also statistically significant among time course by multiple comparison (P=0.000,respectively).By the flow cytometry analysis,it was found that the 100 ng/ml and 400 ng/ml leptin groups could change the distribution of cell cycle of MCF-7 cell line after 48 h. Compared with control group, the cell number decreased by 14.42 ％ in G0/G1 phase (F=10.464, P=0.044),but increased by 7.57 ％ and 22.19 ％ respectively in S phase (F=47.361,P=0.005).The difference was not statistically significant in G2/M phase (F=1.77, P=0.311).However, the effect of apoptosis inhibition was not obvious. Conclusions Leptin could stimulate the proliferation of MCF-7 cell line and change the distribution of cell cycle.But leptin could not inhibit apoptosis of MCF-7 cell line.It suggested that leptin may serve as a risk factor of breast cancer development.
Key words:
Breast neoplasms; Leptin; Cell proliferation; Apoptosis
MR (mineralocorticoid receptor) antagonists have been demonstrated to provide beneficial effects on preventing atrial fibrosis. However, the underlying cellular and molecular mechanisms remain unclear. We aim to determine the role of osteoblast MR in atrial fibrosis and to explore the underlying mechanism. Using osteoblast MR knockout mouse in combination with mutant TGF (transforming growth factor)-β1 transgenic mouse, we demonstrated that MR deficiency in osteoblasts significantly attenuated atrial fibrosis. Mechanistically, MR directly regulated expression of OCN (osteocalcin) in osteoblasts. Both carboxylated and undercarboxylated OCNs (ucOC) were less secreted in osteoblast MR knockout mice. Mutant TGF-β1 transgenic mice supplemented with recombinant ucOC showed aggravated atrial fibrosis. In cultured atrial fibroblasts, ucOC treatment promoted proliferation and migration of atrial fibroblasts, whereas cotreatment with an antagonist for a GPRC6A (G-protein-coupled receptor, family C, group 6, member A) abolished these effects. Western blotting analysis revealed upregulation of PKA (protein kinase A) and CREB (cAMP-response element-binding protein) phosphorylation after ucOC treatment. Inhibition of PKA with its antagonist reduced ucOC-induced proliferation and migration of atrial fibroblasts. Finally, the impact of osteoblast MR deficiency on atrial fibrosis was abolished by ucOC administration in mutant TGF-β1 transgenic mice. Taken together, MR deficiency in osteoblasts attenuated atrial fibrosis by downregulation of OCN to promote proliferation and migration of atrial fibroblasts.
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