Polymeric micelles have exhibited attractive properties as drug carriers, such as high stability in vivo and good biocompatibility, and been successfully used to dissolve various drugs of poor aqueous solubilities. In this study, we developed a new type of polymeric micelles with mannose-mediated targeting and pH-responsive drug release properties for anticancer drug delivery. The polymeric micelles were prepared from an amphiphilic polymer, poly (glycidyl methacrylate)-g-mannose (PGMA-Mannose). An anticancer drug, doxorubicin (DOX), was encapsulated into the micelles during the micellization, and could be released rapidly under acidic condition. The specificity of cellular uptake of the micelles by two different cell lines was studied using confocal laser scanning microscopy and the MTT assay. DOX-loaded micelles were efficiently trapped by mannose-receptor-overexpressing cancer cells MDA-MB-231, whereas mannose- receptor-poor cells HEK293 showed much lower endocytosis towards the micelles under the same conditions. Thus, DOX-loaded micelles displayed higher cytotoxicity to MDA-MB-231 cancer cells as compared with free DOX. The present study demonstrates that PGMA-Mannose micelles are a promising targeted drug delivery system for cancer therapy.
Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Delivery Systems ; HEK293 Cells ; Humans ; Lectins, C-Type ; metabolism ; Mannose ; chemistry ; Mannose-Binding Lectins ; metabolism ; Micelles ; Receptors, Cell Surface ; metabolism

OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.

METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.

RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.

CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.

Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology

AIMTo investigate the role of 1 alpha, 25-dihydroxyvitamin D3 (calcitriol) and its analogues tacalcitol and 24, 25(OH)2D3 on the phagocytosis of human monocyte-derived dendritic cells (MoDC).

METHODSMoDC were generated in vitro by differentiating monocytes in the presence of GM-CSF and IL-4 for 5 days. Expression of mannose receptor (MR) and Fc gamma receptors (Fc gamma Rs) by MoDC was analysed by flow cytometry. Zymosan ingestion was measured to assess the phagocytosis of MoDC.

RESULTSMoDC expressed high level of MR and Fc gamma Rs and showed the capacity of zymosan ingestion. Calcitriol and tacalcitol but no 24, 25(OH)2D3 not only upregulated the expression of MR and Fc gamma Rs on MoDC but also correspondingly enhanced their phagocytosis by increasing zymoasan ingestion. Furthermore, the upregulatory role occurred in the early stage of MoDC differentiation and was irreversible. The upregulatory role of calcitriol was dose dependent.

CONCLUSIONCalcitriol and its analogue tacalcitol may play an important role in dendritic cell binding and capturing foreign antigens at the initiation of immune response.

Calcitriol ; pharmacology ; Calcium Channel Agonists ; pharmacology ; Dendritic Cells ; drug effects ; metabolism ; physiology ; Dihydroxycholecalciferols ; pharmacology ; Humans ; Lectins, C-Type ; metabolism ; Mannose-Binding Lectins ; metabolism ; Monocytes ; cytology ; Phagocytosis ; drug effects ; Receptors, Cell Surface ; metabolism ; Receptors, IgG ; metabolism

OBJECTIVETo explore the mechanism of Flt3 receptor-interacting lectin (FRIL) maintains quiescence of hematopoietic stem cells (HSCs) in vitro.

METHODSCord blood CD34+ cells were cultured in suspension medium supplemented with or without FRIL and FL. Cells were collected at different time points and the expression of some cell cycle regulators, especially those involved in G0/G1 phase regulation were detected on mRNA and protein level.

RESULTSThe expressions of G0/G1 phase related cyclins or CDKs were undetectable in the newly isolated CD34+ cells, expressions of Cyclin D3, CDK6 and P27 were the lowest in FRIL cultured group after 3d's culture (FRIL group: 483 +/- 63, 553 +/- 39, 0.312 +/- 0.030; FL group: 2437 +/- 52, 3209 +/- 98, 0.787 +/- 0.024; BLANK: 914 +/- 105, 1497 +/- 55, 0.616 +/- 0.029, respectively), but the expression of P53 was the highest in FRIL group (FRIL group: 4.476 +/- 0.159; FL group: 0.581 +/- 0.099, BLANK: 2.167 +/- 0.114). The expression of positive regulators of cell cycle in FRIL group were the same as that of FL group and blank group or lower.

CONCLUSIONFRIL preserves HSCs effectively in vitro through the mechanisms of down-regulation of cyclin D3 and CDK6 and activation of P53. P27 is mostly involved in the differentiation of HSCs.

Antigens, CD34 ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Mannose-Binding Lectins ; pharmacology ; Plant Lectins ; pharmacology ; RNA, Messenger ; genetics

Ex vivo maintainance of human stem cells is crucial for many clinical applications. Current culture conditions provide some level support but cytokines induce most quiescent stem cells to proliferate and differentiate. Better control of primitive cells is needed to extend the time and range of manipulation of such cells. A recently identified plant lectin Flt3 receptor-interacting lectin (FRIL) present may a special ability to preserve primitive CB progenitors for extended periods in culture without exogenous cytokines. But the mechanisms of FRIL preserving quiescent primitive cells are still unknown. Recently a novel protein HTm4 and its alternatively spliced variant HTm4S, which serve as hematopoietic cell cycle regulators, have been identified. In this report we studied the effect of FRIL on the in vitro maintenance of quiescent human cord blood stem cells and the expression of the novel hematopoietic cell cycle regulator HTm4 and HTm4S in progenitor cells cultured in FRIL. We analyzed the proliferation and the HPP-CFC proportion of CD34(+) cells treated with FRIL. The human HTm4 and HTm4S mRNA expression was detected by semi-quantitative RT-PCR, and the cell cycle status of CB CD34(+) cells was analyzed by FACS. The results showed that incubation of CD34(+) cells in FRIL resulted in a low proliferation of progenitor cells and fewer cycling cells, but FRIL selectively maintained a higher number of primitive cells with proliferative potential in suspension culture. CB CD34(+) cells cultured in FRIL showed significant diversity in the expression of HTm4 and HTm4S during 0~14 d. On d 0, HTm4 was detected at high level, downregulated on d 1, but upregulated during d 3 to d 14, and reaching the highest level on d 7. But the expression levels of HTm4S changed little in the cells cultured in FRIL except the obviously increased expression on d 7. Exogenous expression showed that HTm4 was localized around the karyon while HTm4S scatted in the cytoplasm, respectively, which may be responsible for their difference in function. Thus, FRIL can preserve quiescent primitive CD34(+), and FRIL's ability to preserve quiescent primitive cells in a reversible manner may significantly expand the time and range of ex vivo manipulations of human stem cells for clinical applications. In other words, HTm4 and HTm4S may play a crucial role in the cell cycle modulation of CD34(+) progenitor cells maintained with FRIL in vitro.
Antigens, CD20 ; biosynthesis ; genetics ; Antigens, CD34 ; metabolism ; Cell Cycle ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Separation ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Mannose-Binding Lectins ; pharmacology ; Membrane Proteins ; biosynthesis ; genetics ; Plant Lectins ; pharmacology

OBJECTIVETo study the mechanism of oligochitosan-induced macrophage activation.

METHODSOligochitosan was chemically modified with fluorophore 2-aminoacridone (2-AMAC). The cellular events of 2-AMAC-oligochitosan-macrophage interaction were analyzed with confocal laser microscopy and the fluorescence intensity of cells was analyzed by BD LSR flow cytometer. The mechanism of oligochitosan uptake by macrophages was studied by competitive inhibition test and the effect of calcium, trypsin and colchicine on oligochitosan recognition and internalization were also determined. RT-PCR was performed to investigate the level of TNF-alpha secretion.

RESULTMacrophage could bind and uptake oligochitosan, which was dependent on the temperature: the uptake proceeded rapidly at 37 degrees C and at 4 degrees C macrophage could only bind oligochitosan. EDTA decreased oligochitosan uptake. Trypsin treatment significantly reduced the internalization, and uptake was recovered by trypsin termination. Colchicine significantly inhibited the internalization process and was dose dependent. 0.1 mol/L mannose inhibited TNF-alpha expression induced by oligochitosan.

CONCLUSIONMacrophage could uptake oligochitosan via mannose receptor mediated pinocytosis. Mannose receptor is crucial for the oligochitosan-induced macrophages activation.

Cells, Cultured ; Chitin ; analogs & derivatives ; pharmacology ; Humans ; Lectins, C-Type ; metabolism ; Macrophage Activation ; drug effects ; Macrophages ; cytology ; Mannose-Binding Lectins ; metabolism ; Pinocytosis ; drug effects ; Receptors, Cell Surface ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

The research progress on Chinese medicine plant resources with pesticide activities, the active components and their reaction mechanism as well as the application and prospect were reviewed in this paper. Some proposals on the exploitation of traditional Chinese medicine plant origin pesticide were given. It is suggested to found compounds with pesticide activities from heat clearing and toxic clearing medicinal plants.
Fungicides, Industrial ; isolation & purification ; pharmacology ; Fusarium ; drug effects ; Insecticides ; isolation & purification ; pharmacology ; Lectins ; isolation & purification ; pharmacology ; Pesticides ; isolation & purification ; pharmacology ; Plant Oils ; isolation & purification ; pharmacology ; Plant Viruses ; drug effects ; Plants, Medicinal ; chemistry

OBJECTIVETo explore the anti-inflammatory mechanisms of high density lipoprotein (HDL) by observing the effects of apoprotein (apo)AI, a major protein component of HDL, on the inflammatory macrophage cell polarity.

METHODSCultured mice marrow-derived macrophages were stimulated with lipopolysaccharide and interferon after 10 µg/ml of apoAI were added to the macrophages for 24 hours. The expression of membrane molecules CD16/32, CD206 were detected by fluorescence activated cell sorting (FACS). ELISA was used to detect the secretion of IL-10 and IL-12. Real-time quantitative PCR was used to detect the mRNA expression of TLR4, MyD88 and IRF5.

RESULTSCompared to macrophages stimulated by interferon and lipopolysaccharide but without pretreatment with apoAI, pre-incubation with apoAI significantly downregulated the expression of CD16/32 (91.17% ± 1.99% vs.50.47% ± 1.02%, P < 0.05), IL-12 [(747.27 ± 3.74)pg/ml vs. (73.80 ± 4.56)pg/ml, P < 0.05], upregulated the expression of CD206(0.33% ± 0.12% vs. 3.00% ± 0.36%, P < 0.05), IL -10 expression [(23.56 ± 4.30) pg/ml vs.(32.91 ± 2.47) pg/ml, P < 0.05], and reduced the mRNA expression of TLR4 (1.000 ± 0.025 vs.0.708 ± 0.003, P < 0.05) , MyD88 (1.591 ± 0.005 vs. 1.341 ± 0.005, P < 0.05) , IRF5 (0.954 ± 0.005 vs. 0.463 ± 0.003, P < 0.05) .

CONCLUSIONApoAI enhances the switch of inflammatory macrophages to anti-inflammatory macrophages possibly through inhibiting TLR4-MyD88-IRF5 pathway.

Animals ; Apolipoprotein A-I ; pharmacology ; Cell Line ; Female ; Interferon Regulatory Factors ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Lectins, C-Type ; metabolism ; Macrophages ; drug effects ; immunology ; metabolism ; Mannose-Binding Lectins ; metabolism ; Mice ; Mice, Inbred C57BL ; Myeloid Differentiation Factor 88 ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, IgG ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo investigate the changes of serum levels of chitinase-3-like-1 protein (YKL-40) and high-sensitivity C-reactive protein (hs-CRP) in patients with non-ST segment elevation acute coronary syndrome (ACS), to explore its correlation with its severity, and to observe the effects of Guizhi Fuling Decoction (GFD) on levels of blood lipids, YKL-40, and hs-CRP.

METHODSRecruited were 72 patients with unstable angina (UA) or non-ST segment elevation myocardial infarction (NSTEMI) at Department of Integrative Medicine, First Affiliated Hospital of Xinxiang Medical College from August 2010 to June 2011. They were randomly assigned to the treatment group (36 cases) and the control group (36 cases). All patients were treated by routine treatment, but patients in the treatment group took GFD additionally. The course of treatment was four weeks. According to the severity degree, all patients were graded to four ranks: low-risk group of UA, medium-risk group of UA, high-risk group of UA, and NSTEMI. The levels of YKL-40 and hs-CRP, and the correlation of severity degree were analyzed. Before and after treatment levels of triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) were measured. Before treatment, at two weeks, and after treatment the serum levels of YKL-40 and hs-CRP were detected. The relationship of YKL-40, hs-CRP and the severity of the disease were analyzed.

RESULTSLevels of YKL-40 and hs-CRP were positively correlated with the severity of the disease respectively (r = 0.729, P < 0.05; r = 0.655, P < 0.05). The positive correlation also existed between YKL-40 and hs-CRP (r = 0.848, P < 0.05). There was no statistical difference in the levels of blood lipids, YKL-40, or hs-CRP between the two groups before treatment (P > 0.05). Compared with before treatment, the levels of YKL-40 and hs-CRP significantly decreased in both groups after two weeks of treatment (P < 0.05). The levels of TG, TC, LDL-C, YKL-40, and hs-CRP significantly decreased, while the HDL-C level increased in both groups after treatment (P < 0.05). The level of HDL-C in the treatment group was higher, while levels of YKL-40 and hs-CRP were lower after treatment, when compared with the control group (all P < 0.05).

CONCLUSIONOn the basis of anti-inflammation and adjusting blood lipids by Western medicine, GFD could further reduce the serum levels of YKL-40 and hs-CRP of ACS patients, elevate the HDL-C level, and play anti-atherosclerosis effects.

Acute Coronary Syndrome ; blood ; drug therapy ; physiopathology ; Adipokines ; blood ; Aged ; C-Reactive Protein ; metabolism ; Chitinase-3-Like Protein 1 ; Cholesterol, HDL ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Lectins ; blood ; Male ; Middle Aged ; Triglycerides ; blood

AIMTo investigate the enhancing effect on insulin absorption through GI tract in mice by using tomato lectin (TL) modified liposomes as the carrier.

METHODSTL-phosphatidylethanolamine (PE) conjugate (TL-PE) was synthesized by using carbodiimide cross-linking method, then the compound was incorporated into the conventional liposomes of insulin. The agglutination test was performed to examine TL biological activities after synthesis and incorporation. When TL modified liposomes were administrated orally to the normal mice or diabetic mice at insulin dose of 350 u x kg(-1), the hypoglycemic effect was determinated according to the blood glucose level.

RESULTSThe blood glucose levels of the normal mice were reduced by modified liposomes. The glucose levels were (85 +/- 5)% at 4 h, (54 +/- 11)% at 8 h, (57 +/- 6)% at 12 h postdose compared with the glucose levels prior to oral administration respectively. However, the conventional liposomes and saline have no hypoglycemic effect. The blood glucose levels of the diabetic mice were obviously reduced by TL modified liposomes, the glucose levels were (38 +/- 13)% at 4 h, (50 +/- 15)% at 8 h, (50 +/- 16)% at 12 h respectively.

CONCLUSIONTL modified liposomes promote the oral absorption of insulin due to the specific-site combination on GI cell membrane.

Administration, Oral ; Animals ; Blood Glucose ; metabolism ; Delayed-Action Preparations ; Diabetes Mellitus, Experimental ; blood ; metabolism ; Drug Carriers ; Drug Delivery Systems ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; pharmacology ; Insulin ; administration & dosage ; pharmacokinetics ; pharmacology ; Intestinal Absorption ; drug effects ; Intestine, Small ; metabolism ; Liposomes ; Mice ; Phosphatidylethanolamines ; chemistry ; Plant Lectins ; chemistry ; pharmacology ; Technology, Pharmaceutical ; methods