Objective To establish a RP-HPLC method for the determination of drug release in vitro of atractylenolide Ⅰ liposomes. Methods The release behavior of the drug from liposomes was studied by the third method for dissolution. ZORBAX C18 column (4.6 mm × 250 mm, 5 μm) was used with a mobile phase of Methanol-Acetonitrile-0.2% from liposomes in vitro fitted the log-normal distribution equation and had a property of sustained release. Conclusion The method is simple, fast and selective. It is suitable for the determination of release profile in vitro of atractylenolide Ⅰ liposomes.

Objective To investigate whether uninduced autologous bone-marrow mononuclear cell (ABM-MNC) could survive and differentiate into myocardial cells and endothelial cells in the infarcted heart. Methods 40 male big-ear Japanese rabbits were divided into two groups randomly: the transplanted group (n=20) and the control group (n=20). The model of acute myocardial infarction was made by left anterior descending artery ligation, which was confirmed by ECG. The cardiac function was evaluated by the echocardiography. 7 days later, BrdU labeled ABM-MNCs were injected into infarcted and marginal area myocardium in the transplanted group, while the control rabbits were injected with saline. 6 weeks later, the hearts were harvested for histology and immunohistochemistry evaluation. Results In the transplanted group, viable cells labeled with BrdU could be identified in the infarcted area, and myocytes and endothelial cells labeled with BrdU can also be found in the border area, these cells demonstrate myogenic differentiation with the expression of α-Actin by immunostaining. Moreover, the vessel density of the transplanted group in the borders of the infarction was higher than the control group (P＜0. 05), but there was no difference in the infarcted areas between two groups (P＞0.05). At the 6 weeks after experiment, the cardiac function was improved in both groups, but the transplanted group improved more than that in the control group (P＜0.05). Conclusion Autologous bone-marrow mononuclear cells injected into the infarcted myocardium could survive in both the infarcted and the border areas, differentiated into endothelial cells and other cells which have obtained the characters of myocytes, and increase the vessel density in border area, improved the cardiac function.

Objective To investigate whether uninduced autologous bone-marrow mononuclear cell (ABM-MNC) could survive and differentiate into myocardial cells and endothelial cells in the infarcted heart. Methods 40 male big-ear Japanese rabbits were divided into two groups randomly: the transplanted group (n=20) and the control group (n=20). The model of acute myocardial infarction was made by left anterior descending artery ligation, which was confirmed by ECG. The cardiac function was evaluated by the echocardiography. 7 days later, BrdU labeled ABM-MNCs were injected into infarcted and marginal area myocardium in the transplanted group, while the control rabbits were injected with saline. 6 weeks later, the hearts were harvested for histology and immunohistochemistry evaluation. Results In the transplanted group, viable cells labeled with BrdU could be identified in the infarcted area, and myocytes and endothelial cells labeled with BrdU can also be found in the border area, these cells demonstrate myogenic differentiation with the expression of α-Actin by immunostaining. Moreover, the vessel density of the transplanted group in the borders of the infarction was higher than the control group (P<0.05), but there was no difference in the infarcted areas between two groups (P>0.05). At the 6 weeks after experiment, the cardiac function was improved in both groups, but the transplanted group improved more than that in the control group (P<0.05). Conclusion Autologous bone-marrow mononuclear cells injected into the infarcted myocardium could survive in both the infarcted and the border areas, differentiated into endothelial cells and other cells which have obtained the characters of myocytes, and increase the vessel density in border area, improved the cardiac function.

BACKGROUND:Calcium sulfate used in kyphoplasty and vertebrolplasty has good physical and chemical properties, exerts no toxic effects on human body and has the degradation performance. But its main drawback is rapid degradation.
OBJECTIVE:To develop a chitosan microsphere with silk fibroin/calcium sulfate cement to prepare drug carrier system.
METHODS:Chitosan microspheres were prepared by the emulsion method. Scanning electron microscopy, particle size analysis and swel ing rate were used to study the properties of the microspheres. Different silk concentrations (3%, 6%and 9%) and weight rates (0.5%,1%and 5%) of chitosan microspheres were used to determine the best formula which has the strongest mechanical properties. The composition of this composite bone cement was detected by using X-ray diffraction and Fourier transform infrared spectroscopy.
RESULTS AND CONCLUSION:When the concentration of silk fibroin was 6%and weight rate of chitosan microspheres was 0.5%, we could obtain the maximum compressive strength, which was (39.17±1.96) MPa. With this composition, the initial setting time was (12.99±1.63) minutes and the final setting time was (21.55±0.54) minutes. The results from X-ray diffraction and Fourier transform infrared spectroscopy demonstrated that the main phase composition was calcium sulfate, and silk and chitosan were also included. The composite chitosan microspheres exhibited a slightly wrinkled surface, but were stil intact in spherical shape, indicating the preparation of chitosan microspheres/silk fibroin/calcium sulfate cement was reliable and the product had good structures and properties.

BACKGROUND:With excelent biocompatibility and osteoconduction, calcium phosphate bone cement has been used in clinic, but the poor mechanical properties and lack of osteoinduction restrict its further use. OBJECTIVE:To investigate the cytocompatibility and cytotoxicity of a novel drug-carrying composite of bone cement composed of chitosan microsphere, α-tricalcium phosphate and silk fibroin. METHODS:MC3T3-E1 cels were cultured in vitroin minimum essential medium alpha medium (α-MEM), which was supplemented with 10% fetal bovine serum, and 1% streptomycin sulfate, extract of the cement material at concentrations of 100% and 50%, and 6.4 mL/L phenol. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to measure cellproliferation and the cytotoxicity was assessed by the activity of lactate dehydrogenase. The MC3T3-E1 cels culturedin vitro were colected and seeded on the composite cement material, and cellmorphology was observed by scanning electron microscope. RESULTS AND CONCLUSION:The extract of composite cement material had no influences on the MC3T3-E1 cellproliferation, showing no obvious cytotoxicity. The scanning electron microscope image showed MC3T3-E1 cels adhered and proliferated wel on the composite cement material composed of chitosan microsphere, α-tricalcium phosphate and silk fibroin, and pseudopodia out of the cels were closely attached to the material surface. In conclusion, the cement composite was proved to have satisfactory cytocompatibility and no obvious cytotoxicity.

BACKGROUND: Vascular injuries to the extremities are frequently concomitant with vascular defects. Vascular transplantation repair can induce infection and vascular occlusion, etc.OBJECTIVE: To analyze the outcome of vascular reconstitution in 44 patients with vascular injuries of the extremities undergoing vascular transplantation.DESIGN: A retrospective case analysis.SETTING: Department of Vascular Surgery and Department of Orthopaedics of Hospital Affiliated to Jinhua College of Profession and Technology.PARTICIPANTS: Forty-four patients with vascular injuries to the extremities undergoing autologous and artificial vascular transplantation were selected at the Department of Vascular Surgery and Department of Orthopaedics of Hospital Affiliated to Jinhua College of Profession and Technology from April 1994 to October 2003. There were 29 patients with open injury and 15 patients with closed injury.METHODS: A total of 52 blood vessels were transplanted into 44 patients, including 42 blood vessels in autologous vein transplantation (35 blood vessels in great saphenous vein transplantation by end-to-end anastomosis, 5 blood vessels in small saphenous vein transplantation by end-to-end anastomosis and 2 blood vessels in superficial femoral vein and popliteal vein transplantation) and 10 blood vessels in artificial and trimming vascular transplantation by interrupted suture technique in end-to-end anastomosis.MAIN OUTCOME MEASURES: Outcomes of autologous and artificial vascular transplantation.RESULTS: Three patients received amputated extremity. Six patients developed ischemic contracture. Seven patients developed imperfect recovery of nerve function. In other patients, blood flow in the graft was satisfactory, and there was good condition of blood circulation at the distal extremities.CONCLUSION: Autologous vein is the first choice in vascular transplantation, and prosthetic material is another choice when necessary. It is important to prevent the occurrence of complication after transplantation such as vascular infection.

Objective To explore the expression of c-fos,bcl-2,bcl-xL gene and apoptosis in rats with diffuse brain injury combination secondary injury(SBI).Methods 40 rats were randomly divided into the normal control group 10,10 cerebral ischemia,head injury group 10 and group 10 SBI to immunohistochemical method to detect brain cells in the c-fos,bcl-2,bcl-xL expression and in-situ to apoptosis(TUNEL).Results The expression of brain injury group and SBI group of cortex area c-fos gene (23.6±9.4),(37.1±5.5)positive cells/entries/H was significantly higher than that of cerebral ischemia group (5.6±1.4) positive cells/entries/H(t= 3.458,t = 3.648, all P<0.01) ;the expression of brain injury group and SBI group cortex area bcl-2 gene (18.2±4.6) ,(15.6±3.7)positive cells/entries/H lower than that brain blood group(23.6±4.3)positive cells/entries/H(t=2.345, t=2.447 ,all P <0.05) ;the expression of bcl-xL gene changes llttle;the apoptosis SBI group cortex area (36.6±5.3)cells/0.1mm2 higher than the brain injury group (21.6±4.4) cells/0.lmm2 (t = 2.378 ,P < 0.05 );the apoptosis and level of bcl-2 expression showed a negative correlation(r = -0.857 ,P <0.01 ).Conclusion The expression of c-fos,bcl-2,bcl-xL were increased with closely related to apoptosis in rats with diffuse brain injury combination secondary injury.

Objective To prepare polypeptide nerve growth factor (NGF) thermosentitive gel and observe drug release in vitro,in order to provide scientific information for biopharmaceuticals delivery system design aiming to treat the inner disease.Methods The thermosensitive in situ gel was prepared with NGF as main component and Pluronic F127 as the gel matrixes.The effect of concentration of gel matrix PF127 on lower critical solution temperature(LCST) were investigated.In vitro release kinetic studies were performed using membraneless dissolution method and reversed-phase high performance liquid chromatography (HPLC) method was adopted to determine NGF content in the dissolution medium.Results The average LCST of NGF-loaded gel prepared by different concentration of PF127 was 28.48-36.26℃ and the gels had good stability.The in vitro release kinetics was well characterized by sustained release and can be fitted by zero order kinetics.The in vitro accumulated release ratio of NGF in the thermosensitive gel reached to above 95％.hβ-NGF loaded in higher PF127 gel resulted in a more sustained release of hβ-NGF from the thermosensitive gel.Conclusion Well-prepared NGF thermosensitive gel is a promising inner ear-oriented drug delivery system for treating deafness and deserves to further development.

Objective To optimize the prescription and preparation technclogy of ligustilide liposomes. Methods Ligustilide liposomes were prepared by ethanol injection method. The encapsulation efficiency was taken as inspection target and the preparation of liposomes was optimized by orthogonal design. HPLC was used to measure the encapsulation efficiency. Results The best prescription was ligustilide-lecithin (1:10),lecithin-cholesterol (2:1),the water phosphate buffer solution (pH 7.5),the hydration temperature was 35 ℃. Conclusion The optimized formulation of ligustilide is reasonable in prescription and practicable in technology.