1.Preparation of podophyllotoxin nanostructured lipid carriers and its effects on immortalized human cervical epithelial cells with HPV infection in vitro.
Shu-bin CHONG ; Kang ZENG ; Guo-feng LI ; Fei REN ; Xiao-liang ZHU ; Jin-jie ZHOU
Journal of Southern Medical University 2011;31(6):1023-1028
OBJECTIVETo study the effect of podophyllotoxin nanostructured lipid carriers (POD-NLC) on immortalized human cervical epithelial cells (H8) infected with HPV in vitro.
METHODSPOD-NLC was prepared by emulsion evaporation method and characterized using transmission electron microscopy, Zetasizer analyzer and high-performance liquid chromatography (HPLC). H8 cells were treated with different concentrations (0.0001-1 µg/ml) of POD-NLC, free POD, or blank nanostructured lipid carriers (NLC), and the cell proliferation was assessed using MTT assay to evaluate the cytotoxic effects. The changes of cell morphology were observed using fluorescence microscopy, and the cell cycle changes and cell apoptosis were analyzed using flow cytometry.
RESULTSPOD-NLC showed a spherical or elliptical shape with good stability in vitro. The average particle size of POD-NLC was 85.6∓10.25 nm, with a Zeta potential of 26.2∓4.1 mV and entrapment efficiency of POD of (88.56∓3.1)%. POD-NLC caused a significant inhibition of H8 cell proliferation in a concentration- and time-dependent manner. At an equivalent concentration, POD-NLC produced a stronger inhibitory effect on cell proliferation than POD. The inhibition rate of H8 cells after a 48-h exposure to POD-NLC and POD reached 95.8% and 65.6%, respectively, and at the highest concentration of 1 µg/ml, the IC(50) of POD-NLC and POD was 0.015 µg/ml and 0.13 µg/ml, respectively. Blank NLC did not obviously affect the proliferation of H8 cells. POD-NLC and POD both caused obvious increases in G(2)/M phase cell percentages and induced typical apoptotic changes of the cells, and their effects were comparable (P>0.05).
CONCLUSIONCompared with POD, POD-NLC has more potent effect in inhibiting H8 cell proliferation and inducing cell apoptosis, suggesting its potential in the treatment of cervical HPV infection.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cervix Uteri ; cytology ; Drug Carriers ; pharmacology ; Epithelial Cells ; drug effects ; Female ; HIV Infections ; pathology ; Humans ; Lipids ; Nanostructures ; Particle Size ; Podophyllotoxin ; chemical synthesis ; pharmacology
2.An experimental study of the inhibitory effects of Tripterygium extract (LLZ) on hypertrophic scar - derived fibroblasts.
Weiguang XIE ; Huiqing JIANG ; Hanbao LI
Chinese Journal of Burns 2002;18(1):32-33
OBJECTIVETo explore the effects of Tripterygium extract (LLZ) on postburn hypertrophic scar - derived fibroblasts, so as to provide us an experimental support for the treatment of postburn hypertrophic scar.
METHODSHypertrophic scar - derived fibroblasts were harvested and cultured in vitro. A serial concentrations of LLZ (5 x 10(-3), 5 x 10(-4), 5 x 10(-5), 5 x 10(-6) g/L) were added to the culture media. The cellular morphology, proliferating activity and the cytotoxicity of the drug were observed after 24 hours of culture.
RESULTSThe fibroblast morphology could be altered by any concentration of LLZ solution. Furthermore, the cell count and the cellular proliferating activity were all decreased obviously by the treatment of LLZ.
CONCLUSIONThe cell morphology and proliferation activity of postburn scar-derived fibroblasts could all be inhibited by LLZ, and it was the result of its toxic effects.
Burns ; drug therapy ; pathology ; Cell Division ; drug effects ; Cell Size ; drug effects ; Cells, Cultured ; Child ; Child, Preschool ; Cicatrix, Hypertrophic ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Fibroblasts ; cytology ; drug effects ; Humans ; Male ; Phytotherapy ; Plant Extracts ; pharmacology ; Tripterygium
3.Roles of paroxetine and corticosterone on adult mammalian ciliary body cell proliferation.
Hua WANG ; Benson W M LAU ; Suk-yu YAU ; Suk-yee LI ; Nelson LEUNG ; Ning-li WANG ; Siu-wa TANG ; Tatia M C LEE ; Kwok-fai SO
Chinese Medical Journal 2010;123(10):1305-1310
BACKGROUNDThe neurogenesis in retina of adult mammals is generally abolished, and this renders the retina lack of regenerative capacity. Despite this, there is a small population of nestin-positive cells in the ciliary epithelium which retains neurogenic potential. The present study aimed at investigating the effect of two drugs, corticosterone and paroxetine, on the cell proliferation of the ciliary body.
METHODSAdult Sprague-Dawley rats were given vehicle, corticosterone, paroxetine, or both corticosterone and paroxetine treatment for 14 days. Cell proliferation in the ciliary body was quantified using 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry. Co-labelling of BrdU and stem cell marker was used to phenotype the BrdU immunoreactive cells.
RESULTSCorticosterone treatment suppressed while paroxetine treatment increased the cell proliferation of the ciliary body. Co-labelling with cell markers revealed that the BrdU positive cells also showed nestin expression but not glial fibrillary acidic protein (GFAP).
CONCLUSIONSThe results illustrate that proliferation of retinal progenitor cells situated in ciliary body are subjected to regulation by selective serotonin reuptake inhibitors (SSRI) and corticosteroid, which is similar to our previous findings in neurogenic regions in central nervous system (CNS). Paroxetine treatment could reverse the suppressive effect of corticosterone on ciliary body cell proliferation. This provides information for future investigation of retinal stem cell biology and potential treatment of retinal degenerative diseases.
Adrenal Glands ; drug effects ; pathology ; Animals ; Body Weight ; drug effects ; Cell Proliferation ; drug effects ; Ciliary Body ; cytology ; drug effects ; Corticosterone ; pharmacology ; Immunohistochemistry ; In Vitro Techniques ; Male ; Organ Size ; drug effects ; Paroxetine ; pharmacology ; Rats ; Rats, Sprague-Dawley
4.Effects of daidzein on sperm quality, testis gain and testosterone in mice.
Zhao-Yu HAN ; Gen-Lin WANG ; Wen CHEN ; Li-Jun ZHANG
National Journal of Andrology 2003;9(8):566-568
OBJECTIVETo evaluate the effects of daidzein on sperm quality of male mice.
METHODSThree different doses of daidzein were supplemented to pubertal male mice for 21 days(5 mg/kg, 100 mg/kg, 1,000 mg/kg ration). The viability of sperm was determined by eosin-Y, the acrosome was observed by Wright-Giemsa staining, and the testosterone was measured by radioimmunoassay.
RESULTSDaidzein at the dose of 5 mg/kg ration significantly increased serum testosterone levels (P < 0.01), prompted testis gain (P < 0.05), and improved spermatozoa quality. Daidzein at dose of 1,000 mg/kg ration could inhibit the secretion of serum testosterone (P < 0.01), without significant variation in spermatozoa quality. Daidzein at dose of 100 mg/kg ration did not significantly affect sperm quality and other index.
CONCLUSIONSDaidzein can affect sperm quality and in dosage-dependant ways.
Animals ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Isoflavones ; pharmacology ; Male ; Mice ; Mice, Inbred Strains ; Organ Size ; drug effects ; Phytoestrogens ; pharmacology ; Spermatozoa ; cytology ; drug effects ; Testis ; drug effects ; Testosterone ; blood
5.Inhibition of all-trans retinoic acid on MDM2 gene expression in astrocytoma cell line SHG-44.
Yi ZENG ; Zhong YANG ; Xiao-Dong LONG ; Chao YOU
Neuroscience Bulletin 2008;24(5):297-304
OBJECTIVETo investigate the impact of all-trans retinoic acid (ATRA) on MDM2 gene expression in astrocytoma cell line SHG-44, and to provide basic data for further research on the progression mechanism and gene therapy of human astrocytoma.
METHODSThe differential expressions of MDM2 gene and protein in SHG-44 cells were detected by cDNA microarray and Western blot, respectively, before and after treatment of ATRA. The expressions of MDM2 protein in WHO grade II and grade IV astrocytomas were determined by immunohistochemical streptavidin-peroxidase method. Some differentially expressed genes were selected randomly for Northern blot analysis.
RESULTSThe intensity ratio of ATRA-treated to untreated SHG-44 cell was 0.37 in the cDNA microarray, suggesting that the expression of MDM2 gene was down-regulated in SHG-44 cells after treatment with ATRA. Some genes differentially expressed in the microarray were confirmed by Northern blot. Western blot demonstrated that the optical density ratios of MDM2 to beta-actin in ATRA-treated and untreated SHG-44 were 14.02+/-0.35 and 21.40+/-0.58 (t = 24.728, P = 0.000), respectively, suggesting that the expression of MDM2 protein was inhibited in ATRA-treated SHG-44 cells. Moreover, the percentages of MDM2-positive protein were 24.00% (6/25) and 56.52% (13/23) (chi(2) = 5.298, P = 0.021) in WHO grade II and grade IV astrocytomas, respectively, suggesting that the expression of MDM2 protein may increase along with the elevation of astrocytoma malignancy.
CONCLUSIONATRA can inhibit MDM2 gene expression in SHG-44 cells, and MDM2 is related to astrocytoma progression.
Antineoplastic Agents ; pharmacology ; Astrocytoma ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Size ; drug effects ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism ; Time Factors ; Tretinoin ; pharmacology
6.Preparation and evaluation of RGD and TAT co-modified paclitaxel loaded liposome.
Journal of Central South University(Medical Sciences) 2014;39(8):769-774
OBJECTIVE:
To prepare Arg-Gly-Asp (RGD) and cell penetrating peptide TAT co-modified paclitaxel loaded liposome (RGD/TAT-LP-PTX) for MCF-7 cell inhibition.
METHODS:
The co-modified liposome was prepared by film-ultrasonic method. The appearance, particle size and zeta potential were evaluated. The cellular uptake by MCF-7 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/TAT-LP-PTX was evaluated by MTT assay. Tumor spheroids were used to evaluate anti-tumor ability of RGD/TAT-LP-PTX in vitro.
RESULTS:
The particle diameter of the co-modified liposome was (138.8 ± 12.4) nm with the Zeta potential of (25.85 ± 2.75) mV. The entrapment efficiency of PTX was 88.3%. The RGD/TAT-LP uptaken by MCF-7 cells at 4 h was 1.79 times higher than that at 2 h. The co-modified liposome uptaken by MCF-7 cells was 2.25 and 2.72 times higher than that of TAT-LP and RGD-LP, respectively. The anti-proliferation rate of RGD/TAT-LP-PTX increased with time. The inhibition rate of RGD/TAT-LP-PTX for MCF-7 cells at 48 h was 1.78 times higher than that at 24 h. The MTT assay demonstrated the cell viability of RGD/TAT-LP-PTX was 1.65, 1.82 and 2.55 times higher than that of TAT-LP-PTX, RGD-LP-PTX and LP-PTX, respectively.
CONCLUSION
Co-modified liposome may serve as a promising breast cancer delivery system for antitumor drugs.
Antineoplastic Agents
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pharmacology
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Breast Neoplasms
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Cell Survival
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Humans
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Liposomes
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MCF-7 Cells
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drug effects
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Oligopeptides
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chemistry
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Paclitaxel
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pharmacology
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Particle Size
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Peptide Fragments
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chemistry
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Spheroids, Cellular
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drug effects
7.In Utero Exposure to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin Affects the Development of Reproductive System in Mouse.
Mei Hua JIN ; Hae Kyung KO ; Chang Hee HONG ; Sang Won HAN
Yonsei Medical Journal 2008;49(5):843-850
PURPOSE: Exposure of male reproductive organs to 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) has been reported to cause developmental changes. In this study, we evaluated the effects of in utero TCDD exposure on male reproductive development. MATERIALS AND METHODS: Pregnant C57BL/6 mice were administered a single intraperitoneal injection of TCDD (1microgram/kg) on gestation day (GD) 15. The offspring were examined in the immature stage on postnatal day (PND) 30 and in the mature stage on PND 60. The testes were examined for histological changes, androgen receptor (AR), proliferating cell nuclear antigen (PCNA) and apoptosis following the measurement of morphological changes. RESULTS: Anogenital distance (AGD) and testis weights were reduced by TCDD exposure both on PND 30 and PND 60 while body weights and length of male offspring were not affected by TCDD. The regular sperm developmental stage was impaired with TCDD treatment on PND 30. However, no difference was found between the control group and TCDD groups on PND 60. Simultaneously, the expression of AR was also reduced on PND 30, while it was increased on PND 60 compared with the control group. The expression of PCNA was decreased whereas apoptosis was not affected by TCDD both on PND 30 and PND 60. CONCLUSION: These results suggest that in utero exposure to TCDD influences the development of testes by inhibiting the expression of AR and PCNA. Moreover, the adverse effects of TCDD on male offspring reduced over time.
Animals
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Apoptosis/drug effects
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Cell Proliferation
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Embryonic Development/*drug effects
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Environmental Pollutants/*toxicity
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Female
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Male
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*Maternal Exposure
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Mice
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Mice, Inbred C57BL
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Organ Size/drug effects
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Pregnancy
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Receptors, Androgen/metabolism
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Testis/*drug effects/embryology/pathology
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Tetrachlorodibenzodioxin/*toxicity
8.Effect of oviductus ranae and oviductus ranae eggs on bone metabolism and osteoporosis.
Dan-Hui WANG ; Wei WU ; Jian-Ming TIAN ; Zhao-Hui WANG ; Dan-Tong WANG ; Ke XIANG ; Guo-Ying ZHU ; Tie HAN
Chinese journal of integrative medicine 2013;19(7):532-538
OBJECTIVETo evaluate the roles or effects of oviductus ranae (OR) or oviductus ranae eggs (ORE) in preventing and treating postmenopausal osteoporosis.
METHODSIn vivo experiment: Sixty female adult Wistar rats were randomly divided into 5 groups of 12. To provide an osteoporosis model 4 groups of rats were ovariectomized (OVX), with the 5th being sham operated. Medication commenced 7 days after the operation and lasted continuously for 12 weeks. Sham operated and OVX groups were given equivalent volumes of 5% Tween-80. The other three groups intragastrically received conjugated estrogens (CE), OR or ORE of the corresponding doses. At the 12th week, serum estrogen, bone gla protein (BGP), serum calcium, phosphorus, and alkaline phosphatase (ALP) were assayed; bone mineral densities (BMD) were measured and bone scanning was conducted; uteri were weighed, and weight, volume and length of the femoral bones were determined; and cortical thickness of femoral heads and area of bone trabecula were measured by image analyzer. In vitro experiment: Eighty 10-month old SD rats, with equal numbers of males and females, were randomly divided into 8 groups. Osteoblasts were isolated from neonatal rat calvariae, and the cells were exposed to various concentrations of serum from OR and ORE groups to study the impact of these sera on osteoblastic proliferation, ALP activity and mineralization. Osteoclastic numbers were determined using tartrate resistant acid phosphatase (TRAP).
RESULTSIn vivo experiment: The body weight of the four OVX groups increased significantly (P<0.01). Uterine weight of the CE group was the highest (P<0.01); Compared with the model group, estrogen level, BMD, bone scanning/bone imaging index weight of the femoral bones, cortical thickness of femoral heads in the OR and ORE groups increased significantly (P<0.05, P<0.01); femoral volume in the ORE group increased significantly (P<0.05); and the content of osteocalcin, phosphorus, and ALP in serum decreased significantly (P<0.05, P<0.01). In vitro experiment: Sera from OR and ORE groups had notable effects on the proliferation of osteoblasts (P<0.05 and P<0.01, repsectively) and stimulated the formation of calcium nodes (P<0.05, P<0.01), while the enhancement of ALP activity in osteoblasts was significant (P<0.05, P<0.01). The number of TRAP-positive cells was significantly reduced as well (P<0.01).
CONCLUSIONSOR and its eggs could effectively suppress OVX-induced osteoporosis in rats, and increase bone turnover possibly by both an increase in osteoblastic activity and a decrease in osteoclastic activity. The present study provides evidence that OR and its eggs could be considered a complementary and alternative medicine for the treatment of postmenopausal osteoporosis.
Acid Phosphatase ; metabolism ; Alkaline Phosphatase ; metabolism ; Animals ; Biomarkers ; blood ; Body Weight ; drug effects ; Bone Density ; drug effects ; Bone and Bones ; metabolism ; Calcification, Physiologic ; drug effects ; Cell Count ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Female ; Femur ; drug effects ; metabolism ; pathology ; Isoenzymes ; metabolism ; Male ; Materia Medica ; pharmacology ; therapeutic use ; Organ Size ; drug effects ; Osteoblasts ; drug effects ; enzymology ; pathology ; Osteoclasts ; drug effects ; enzymology ; pathology ; Osteoporosis ; blood ; drug therapy ; metabolism ; physiopathology ; Ovariectomy ; Ovum ; metabolism ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; Uterus ; drug effects ; pathology
9.CD99 type II is a determining factor for the differentiation of primitive neuroectodermal cells.
Eun Jung LEE ; Hyun Gyu LEE ; Sung Hye PARK ; Eun Young CHOI ; Seong Hoe PARK
Experimental & Molecular Medicine 2003;35(5):438-447
CD99 is a 32-kDa cell surface molecule present on thymocytes, peripheral T cells, many other hematopoietic stem cells and somatic cells were implicated in cell-cell adhesion and cell-activation phenomena. Two major subtypes have been identified so far, designated CD99 type I and type II. We have investigated the correlation between the degree of neural differentiation and the expression of CD99 subtypes in three differentially differentiated cell lines such as CADO-ES1, RD-ES, and SH-N-SY5Y, in order of differentiation. In addition, we induced differentiation of the RD-ES cell line by N(6),2'-dibutyryl-cAMP (db-cAMP). Six days after treatment with db-cAMP, RD-ES cell line has changed its morphology from uniform round cells to cells with neurites, and initially CD99 type II-overexpressed RD-ES cells showed significant down-regulation of CD99 type II, whereas CD99 type I expression remained constant. When RD- ES cells were transfected with the cDNA encoding for CD99 type I-green fluorescence protein (GFP) and type II-GFP, CD99 type II transfected RD-ES cell line remained unchanged with morphology of undifferentiated form. Our data suggest that CD99 type II acts as a negative regulator in the neural differentiation of precursor cells that might occur during nerve system development.
Antigens, CD/genetics/*metabolism
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Bucladesine/pharmacology
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Cell Adhesion Molecules/genetics/*metabolism
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*Cell Differentiation/drug effects
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Cell Line
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Cell Size/drug effects
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Ectoderm/*cytology/drug effects/*metabolism
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Human
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Neurites/drug effects
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Neurons/*cytology/drug effects/*metabolism
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Protein Isoforms/genetics/metabolism
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Support, Non-U.S. Gov't
;
Transfection
10.Effect of phospholipid composition on pharmaceutical properties and anti-tumor activity of stealth liposomes containing brucine.
Minglei CHEN ; Jun CHEN ; Ting HOU ; Yun FANG ; Weiwei SUN ; Rongrong HU ; Baocang CAI
China Journal of Chinese Materia Medica 2011;36(7):864-867
OBJECTIVETo compare the pharmaceutical properties and the anti-tumor activities of three kinds of stealth liposomes prepared with different phospholipid composition containing brucine.
METHODStealth liposomes with different phospholipids composition, such as soybean phosphatidycholine (SPC), hydrogenated soybean phosphatidylcholine (HSPC) and the complex of SPC and HSPC, were prepared by ammonium sulfate transmembrane gradient method. Pharmaceutical properties such as shape, encapsulation efficiency and size of three stealth liposomes were compared intensively. Anti-tumor activity of SPC, HSPC and novel stealth liposomes composed of both SPC and HSPC were compared by established mouse liver cancer H22 model. Meanwhile, the mice body weight and immune organ weight were also compared.
RESULTThe encapsulation efficiency of novel, SPC and HSPC stealth liposomes were 77.7%, 64.8% and 74.8%, respectively. The mean diameters of them were less than 100 nm. The tumor inhibition rate of novel, HSPC and SPC stealth liposomes were 57.88%, 49.15%, 23.37%, respectively. The mice body weight, thymus gland index of three stealth liposomes group and spleen index of novel stealth liposomes group had no significant difference with the negative group while SPC and HSPC stealth liposomes group increased the spleen index.
CONCLUSIONPhospholipids composition is the key factor which determines the antitumor activity of brucine-loaded stealth liposomes.
Animals ; Antineoplastic Agents ; chemistry ; pharmacology ; Body Weight ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Liposomes ; adverse effects ; chemistry ; Mice ; Particle Size ; Phospholipids ; chemistry ; Strychnine ; analogs & derivatives ; chemistry