1.Preparation of doxorubicin-loaded nanoparticles based on modified popaccharide and its targeting effect on hepatocellular carcinoma cells
Yang BAI ; Qingqing XIONG ; Hai WANG ; Liyun PANG ; Tianqiang SONG
International Journal of Biomedical Engineering 2017;40(1):-
Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)% and the encapsulation efficiency was (64.2±0.9)%.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.
2.Effects of different types of cell culture dishes on fluorescent image with confocal microscope
Liyun PANG ; Hai WANG ; Huanlai LIU ; Yuqi XU ; Dunwan ZHU
International Journal of Biomedical Engineering 2012;35(3):177-180,后插5,后插6
Objective To investigate the effects of different thickness of cell culture dishes on fluorescent image with confocal microscope.Methods The fluorescent staining experiments of live cells and fixed cells were used to determine the differences among three dishes with different thickness coverslips of 0.085~0.13 mm,0.13~0.16 mm and 0.16~0.19 mm,while the cell appearance,fluorescence lightness and mean of fluorescence intensity were studied with confocal microscope.Results Demonstrated by the results of cytoskeleton staining experiments,the dish with 0.13~0.16 mm thickness coverslip was the best choice for confocal microscope,the dish with 0.16~0.19 mm thickness coverslip was the second one,the dish with 0.16~0.19 mmthickness coverslip was the last one.ConclusionThe dish with 0.13~0.16 mm thickness coverslip is the best choice for confocal microscope.On this type of dish,the cytoskeleton is unfolding and clear after staining.The intensity of fluorescence is the strongest,and the imaging effect is the best.
3.Heme oxygenase decreases the generation of ROS in AngⅡ induced proliferation and hypertrophy of cultured vascular smooth muscle cells
Dongwei PANG ; Li HAN ; Jie LIU ; Tiemin MA ; Hai XU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM:To investigate the role of heme oxygenase (HO) in AngⅡ induced proliferation and hypertrophy of cultured vascular smooth muscle cells. METHODS:(1) Western blotting analysis was carried out to detect protein level of HO-1 in the tissues. (2) [3H]-TdR,[3H]-leucine incorporation was measured in cultured vascular smooth muscle cells. (3) 2',7'-dichlorofluorescin diacetate (DCFH-DA) as an index was used to determine the cellular reactive oxygen species (ROS) level. RESULTS:(1) No significant difference in HO-1 protein expression level between AngⅡ-stimulated and control groups was observed,but HO-1 protein level in Hemin-induced group was higher than that in other two groups (P
4.Changes of HO/CO pathway in renal hypertension of two-kidney one-clip hypertensive rats
Dongwei PANG ; Li HAN ; Junchang LIU ; Tiemin MA ; Hai XU
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To investigate the effect of heme oxygenase /carbon monoxide(HO/CO) system on the pathogenesis of hypertension and effect of Heme-L- Lysinate(HLL), Zine protoporphyrin-IX(ZnPPIX) on vascular tension in a two kidney one-clip model. METHODS: The changes in mean arterial pressure and heart rate as well as phenylephrine(PHE)-induced contraction in isolated aortic rings of HLL, ZnPPIX- treated rats were examined after intraperitoneal administration of HLL and carbon monoxide( CO). RESULTS: (1) Injection of exogenous CO resulted in a significant decrease in mean arterial pressure in Sham and 2K1C groups(P
5.Effect of heme oxygenase on vascular remodeling in renal hypertension
Li HAN ; Dongwei PANG ; Tiemin MA ; Hai XU ; Lilin WU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To investigate the effect of heme oxygenase on vascular remodeling in renal hypertension. METHODS: Male Wistar rats were randomly divided into sham-operated, 2K1C (two-kidney one-clip) and hemin-induced groups. Four weeks after the treatments, the thickness of aortic media and HO enzymatic activity of the aorta were determined. Immunohistochemical staining was carried out to detect protein of HO-1 in the aorta. RESULTS: The blood pressure in 2K1C renal hypertension rats started to increase two weeks after the surgery and stabled at a high level at the 4th week. Hemin, an inducer of HO-1, markedly inhibited the increase in blood pressure. Aortic medium thickness of the 2K1C rats at 4th week was 27 5% thicker than that in the sham-operated rats. The thickness of aortic medium of the hemin-induced rats was 16 1% less than that in 2K1C group. At the 4th week after operation, protein level and enzymatic activity of HO-1 in aorta were higher than that in 2K1C group compared to those in the sham-operated group. CONCLUSION: Renal hypertension caused vascular remodeling and the activation of HO-1. HO-1 induction decreased the blood pressure of renal hypertension and reduced vascular remodeling.
6.Alpha-lipoic acid inhibits mitochondrial oxidative stress in the rat skeletal muscle with chronic hypoxia exposure
Pin XIAO ; Yihui PANG ; Peng PENG ; Hai BO
Chinese Journal of Tissue Engineering Research 2013;(28):5216-5222
BACKGROUND: α-lipoic acid is named as “nature antioxidant” and “mitochondrial nutrition”. But it is unclear whether α-lipoic acid can be used to protect skeletal muscle with chronic hypoxia exposure, as wel as the relative mechanism. OBJECTIVE: To observe the effect of α-lipoic acid on the antioxidant enzymes and oxidative stress in rat skeletal muscle with chronic hypoxia exposure, and to investigate the relative signaling pathway of α-lipoic acid. METHODS: Thirty-six Sprague Dawley rats were randomly divided into three groups: normoxia control group, hypoxia control group, and hypoxia+α-lipoic acid group. Rats in the hypoxia control group were subjected to hypoxia exposure in normobaric hypoxic tent with 11.3% oxygen concentration. Rats in the hypoxia+α-lipoic acid group were induced by adding α-lipoic acid (0.25%) in the normal diet. Al the interventions were lasted for 4 weeks. RESULTS AND CONCLUSION: α-lipoic acid in hypoxia could markedly enhance the mitochondrial Sirtuin-3 expression, improve the mitochondrial adenosine triphosphate synthesis activity and membrane potential, up-regulate the mitochondrial state 3 respiratory rate, respiratory control ratio and ratio of phosphorus to oxygen, down-regulate the mitochondrial state 4 respiratory rate and promote and up-regulate the activity of mitochondrial antioxidant enzymes such as manganese superoxide dismutase, glutathione peroxidase and catalase, thus inhibiting mitochondrial H2O2 generation rate and reducing mitochondrial malondialdehyde level. The results indicated that α-lipoic acid could improve the efficiency of energy metabolism of chronic hypoxia skeletal muscle mitochondria and inhibit reactive oxygen generation, and it could inhibit the oxidative stress through improving antioxidant enzyme activity of mitochondria. The protection mechanism of α-lipoic acid on hypoxia skeletal muscle mitochondria may be related to the increasing of mitochondrial state 3 respiratory rate.
7.Observation on curative effect of puerarin on Alzheimer's disease in ovariectomized versus non-ovariectomized rat models
Hai LI ; Guangfu PANG ; Jisheng XIE ; Biao LI ; Shuqui ZHANG
Chinese Journal of Geriatrics 2013;32(10):1110-1112
Objective To observe the curative effect of puerarin on Alzheimer's disease (AD) in ovariectomized versus non-ovariectomized rat.Methods 100 6-month-old female SD rats were randomly divided into 5 groups:the control group,non ovariectomized AD group,ovariectomized AD group,non-ovariectomized AD treatment group and ovariectomized AD treatment group (n =20each).Rat model of AD was made by treating with combined D-galactose and sodium nitrite for 60days.Treatment group rats were treated with puerarin for 8 weeks.At the end of the experiment,rats learning and memory abilities were tested by water maze,the serum estradiol,progesterone,follicle stimulating hormone and luteinizing hormone were measured,and the activities of cholinesterase (TchE) and superoxide dismutase (SOD),malondialdehyde (MDA) concentration in brain homogenate were determined.Results Compared with the control group,TchE activity and MDA content were increased,and SOD activity was reduced (all P<0.01),and learning and memory abilities were significantly decreased in the ovariectomized and non-ovariectomized AD groups.Compared with the model groups,the activity of TchE and MDA concentration were reduced,and SOD activity was increased (all P<0.01),and learning and memory abilities were significantly improved in the two treatment groups.As compared with non ovariectomized AD treatment group,the increase in SOD and TchE activities and decrease in MAD concentration were much more in ovariectomized AD treatment group (P<0.05),and the improvement in learning and memory abilities was more significant.Conclusions Puerarin has a therapeutic effect on rat model of AD,even better on the ovariectomized rat model of AD.
8.Detection the serum IgM and IgG specific for TORCH in 1307 women in the period of pre-pregnancy and pregnancy.
Wen QIN ; Da-chun HU ; Hai-li PANG
Chinese Journal of Experimental and Clinical Virology 2011;25(4):292-294
OBJECTIVEThe objective is to obtain the information of women in the period of prepregnancy and pregnancy infected by TORCH (Toxoplasma, Rabella Virus, Cytomegalovirus, and Herpes Simplex Virus) in Kunming for preventing fetus from infection by TORCH in uterus.
METHODSThe serum IgM and IgG specific for TORCH from 1307 women in the period of pre-pregnancy and pregnancy were measured with ELISA and The reagent is the product of virion/serion.
RESULTSThe serum IgM specific for TORCH were detected in 2.83% of 1307 pregnant women for Toxoplasma, 2.37% for Rebulla Virus, 0.46% for Cytomegalovirus, 2.45% for Herpes Simplex Virus. The total positive rate of serum IgM specific for TORCH was 1.45%. The serum IgG specific for TORCH were detected in 3.98% of 1307 pregnant women for Toxoplasma, 72.3% for Rebulla Virus, 97.78% for Cytomegalovirus, 80.34% for Herpes Simplex Virus. The total positive rate of serum IgG specific for TORCH was 63.60%. There was no significant difference among the women with different pregnant situation in terms of the serum IgM and IgG specific for TORCH.
CONCLUSIONThere are some of women in the period of pre-pregnancy and pregnancy with current infection by TORCH in Kunming. It is recommended to screen for infection by TORCH in women in the period of pre-pregnancy and pregnancy to prevent fetus from infection by TORCH in uterus.
Adolescent ; Adult ; Antibodies, Protozoan ; blood ; Antibodies, Viral ; blood ; Cytomegalovirus ; immunology ; isolation & purification ; Female ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Mass Screening ; Middle Aged ; Pregnancy ; Pregnancy Complications, Infectious ; immunology ; parasitology ; prevention & control ; virology ; Rubella virus ; immunology ; isolation & purification ; Simplexvirus ; immunology ; isolation & purification ; Toxoplasma ; immunology ; isolation & purification ; Women ; Young Adult
9.Enlightenment of adverse reaction monitoring on safety evaluation of traditional Chinese medicines.
Hai-bo SONG ; Xiao-xi DU ; Jing-tian REN ; Le YANG ; Xiao-xin GUO ; Yu PANG
China Journal of Chinese Materia Medica 2015;40(8):1620-1623
The adverse reaction monitoring is important in warning the risks of traditional Chinese medicines at an early stage, finding potential quality problems and ensuring the safe clinical medication. In the study, efforts were made to investigate the risk signal mining techniques in line with the characteristics of traditional Chinese medicines, particularly the complexity in component, processing, compatibility, preparation and clinical medication, find early risk signals of traditional Chinese medicines and establish a traditional Chinese medicine safety evaluation system based on adverse reaction risk signals, in order to improve the target studies on traditional Chinese medicine safety, effective and timely control risks and solve the existing frequent safety issue in traditional Chinese medicines.
China
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epidemiology
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Drug Evaluation
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Drug-Related Side Effects and Adverse Reactions
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epidemiology
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etiology
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Drugs, Chinese Herbal
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adverse effects
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therapeutic use
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Humans
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Product Surveillance, Postmarketing
10.Opportunity and challenge of post-marketing evaluation of traditional Chinese medicine.
Xiao-Xi DU ; Hai-Bo SONG ; Jing-Tian REN ; Le YANG ; Xiao-Xin GUO ; Yu PANG
China Journal of Chinese Materia Medica 2014;39(18):3427-3429
Post-marketing evaluation is a process which evaluate the risks and benefits of drug clinical application comprehensively and systematically, scientific and systematic results of post-marketing evaluation not only can provide data support for clinical application of traditional Chinese medicine, but also can be a reliable basis for the supervision department to develop risk control measures. With the increasing demands for treatment and prevention of disease, traditional Chinese medicine has been widely used, and security issues are also exposed. How to find risk signal of traditional Chinese medicine in the early stages, carry out targeted evaluation work and control risk timely have become challenges in the development of traditional Chinese medicine industry.
Drug Evaluation
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methods
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Drugs, Chinese Herbal
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Humans
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Medicine, Chinese Traditional