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.

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.

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

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

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.

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.

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.

Objective To investigate the cell compatibility of polystyrene(PS) plate chemically modified with RGD peptides.Methods PS surfaces were carboxylated by permanganate oxidation in diluted sulfuric acid,and carboxyls were activated with water-soluble carbodiimide to graft with gelatin,collagen and RGD peptides.IR,X-ray photo-electronic spectroscopy (XPS) and dynamic contact angle were used to characterize the surface modification of PS surface.Results XPS results confirmed the existence of nitrogen element from protein molecules and the covalently binding of proteins to PS surfaces.Dynamic contact angle measurement indicated hydrophilicity of PS surfaces was improved obviously after grafting modification.The cell culture results showed that the cell adhesion and proliferation was better on modified surfaces than the initial.Conclusion The cell compatibility of PS surface was great improved after modification with RGD peptides,which would provide a potential strategy to improve the culture of purified endothelial progenitor cells isolated by immunomagnetic beads.

BACKGROUND: Deep vein thrombosis (DVT) of the lower limbs is one of the severe complications of total hip replacement (THR)during the perioperative period. The incidence rate was high. The effect of primary disease on the DVT after THR in the elderly remains poorly understood.OBJECTIVE: To explore the effect of primary disease on DVT after THR in the elderly.METHODS: 147 cases with unilateral THR aged 64-93 years, were included and divided into two groups. Fracture group contained 68 cases, which of them were all traumatic femoral neck fracture ones. Osteopathia group contained 79 cases, and they had no traumatic injury. We selected total hip prostheses according to their physiological age, preoperative socialization ability,substantia ossea and life expectancy. Biological prosthesis in 5 cases and mixed prosthesis in 12 cases were used, while the others used bone cement prosthesis. If the patient had pain and/or swelling on the injured limb, with or without Homans/Neuhofs sign, we did the pressurized ultrasonic Doppler to examine whether the patient had got DVT.RESULTS AND CONCLUSION: In the fracture group, 32 cases had swelling in the injured limbs, and 20 cases with pain, 15 cases with Homans/Neuhofs sign, and 29 DVT cases were confirmed by the pressurized ultrasonic Doppler. One femoral neck fracture case of THR had no DVT clinical signs and was dead 17 days later. The autopsy found that it was an mixed type combined of pulmonary embolism; Osteopathia group: 20 cases had swelling in the injured limbs, and 11 cases of pain, 9 cases of Homans/Neuhofs sign, 20 DVT cases were confirmed by the pressurized ultrasonic Doppler. The blood coagulation state was greater, and the incidence rate of DVT in the lower limbs was greater in the fracture group compared with osteopathia group (P ＜ 0.05). These indicated that femoral neck fracture is a high risk factor for DVT development in lower limb affer THR in the elderly.

Objective:To explain the mechanism of benefiting qi and activating blood formula on chronic heart failure (CHF)by observing the signal transduction through the medium of AngⅡ,PKC in rat myocardium.Methods:Healthy male SD rats were selected and CHF model were made by ligation of coronary artery combined with starvation and exhausted swimming. Successful model rats were randomly divided into 4 groups:model group,Chinese medicine high dose group,Chinese medicine middle dose group,west medicine control group,each group with 8 rats.In addition,normal control group was set.After 8 weeks, the content of AngII,PKC in myocardial tissue was tested by ABC immunohistochemical method,and the ultrastructural changes of myocardial cell were observed with transmission electronic microscopy.Results:Compared with normal control group,heart function decreased obviously(P