Covalent modification of bovine testicular hyaluronidase with chondroitin sulphate led to changes in the pattern of glycation of native and modified enzyme in its reaction with neutral saccharides and N-acetylhexosami...Covalent modification of bovine testicular hyaluronidase with chondroitin sulphate led to changes in the pattern of glycation of native and modified enzyme in its reaction with neutral saccharides and N-acetylhexosamines. Thus, mono- and di-saccharides inactivated the native hyaluronidase to a greater extent than the chondroitin sulfate-modified enzyme. N-acetylhexosamine, on the opposite, inactivated the modified hyaluronidase to a greater extent than the native one. These properties made it possible to use native and modified hyaluronidase as an informative research system for in vivo measurement of the predominant type of saccharide agents in the circulation. The proposed approach was experimentally substantiated by obtained results of the study on these interactions of hyaluronidase derivatives with hyaluronan fragments and their mixture. In a model of post-ischemic perfusion of the rat limb, the effect of hyaluronidase derivatives and their components on restoration of the microcirculation were tracked using laser Doppler flowmetry. Native hyaluronidase accelerated the restoration of initial level of microcirculation, but modified enzyme was markedly inhibited by glycocalyx degradation products. N-acetylhexosamine was positioned at the reducing terminal of these products as a natural label for these glycocalyx fragments. These and other data obtained under various experimental conditions supported the participation of endothelial glycocalyx in microcirculation disturbances.展开更多
In this study,the macroporous calcium phosphate cement with oriented pore structure was prepared by freeze casting.SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected ...In this study,the macroporous calcium phosphate cement with oriented pore structure was prepared by freeze casting.SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected aligned along the ice growth direction.The porosity of the as-prepared porous CPC was measured to be 87.6% by Archimede's principle.XRD patterns of specimens showed that poorly crystallized hydroxyapatite was the main phase present in the hydrated porous calcium phosphate cement.To improve the mechanical properties of the CPC scaffold,the 15% gelatine solution was infiltrated into the pores under vacuum and then the samples were freeze dried to form the CPC/gelatine composite scaffolds.After reinforced with gelatine,the compressive strength of CPC/gelatin composite increased to 5.12 MPa,around 50 times greater than that of the unreinforced macroporous CPC scaffold,which was only 0.1 MPa.And the toughness of the scaffold has been greatly improved via the gelatine reinforcement with a much greater fracture strain.SEM examination of the specimens indicated good bonding between the cement and gelatine.In conclusion,the calcium phosphate cement/gelatine composite with oriented pore structure prepared in this study might be a potential scaffold for bone tissue engineering.展开更多
Objective： To testify the effect of the stem cells derived from the widely distributed fat tissue on repairing full-thickness hyaline cartilage defects. Methods： Adipose-derived stem cells （ADSCs） were derived fro...Objective： To testify the effect of the stem cells derived from the widely distributed fat tissue on repairing full-thickness hyaline cartilage defects. Methods： Adipose-derived stem cells （ADSCs） were derived from adipose tissue and cultured in vitro. Twenty- seven New Zealand white rabbits were divided into three groups randomly. The cultured ADSCs mixed with calcium alginate gel were used to fill the full-thickness hyaline cartilage defects created at the patellafemoral joint, and the defects repaired with gel or without treatment served as control groups. After 4, 8 and 12 weeks, the reconstructed tissue was evaluated macroscopically and microscopically. Histological analysis and qualitative scoring were also performed to detect the outcome.Results： Full thickness hyaline cartilage defects were repaired completely with ADSCs-derived tissue. The result was better in ADSCs group than the control ones. The microstructure of reconstructed tissue with ADSCs was similar to that of hyaline cartilage and contained more cells and regular matrix fibers, being better than other groups. Plenty of collagen fibers around cells could be seen under transmission electron microscopy. Statistical analysis revealed a significant difference in comparison with other groups at each time point （t=4.360, P〈0.01）. Conclusion： These results indicate that stem cells derived from mature adipose without induction possess the ability to repair cartilage defects展开更多
Objective: To explore the possibility of repairi ng long segmental bone defects and preventing infection with cefazolin loaded bo ne matrix gelatin (C-BMG).Methods: C-BMG was made from putting cefazolin into BMG by va...Objective: To explore the possibility of repairi ng long segmental bone defects and preventing infection with cefazolin loaded bo ne matrix gelatin (C-BMG).Methods: C-BMG was made from putting cefazolin into BMG by vac uum absorption and lyophilization techniques. The sustaining period of effective drug concentration in vitro and in vivo was detected. The time of inhibiting ba cteria,and the drug concentration in local tissues (bone and muscle) and plasma after implantation of C-BMG were examined by high performance liquid chromatog raphy. Results: The effective inhibition time to staphylococcus aureus of C-BMG was 22 days in vitro; while 14 days in vivo. The cefazolin concentrat ion in local tissues was higher in early stage,and later it kept a stable and l ow drug release. C-BMG showed an excellent ability to repair segmental long bon e defects.Conclusions: C-BMG can gradually release cefazolin with effect ive drug concentration and has excellent ability to repair segmental bone defect s. It can be used to repair segmental long bone defects and prevent infection af ter operation.展开更多
OBJECTIVE: To investigate the effects of Zibushenjing Fang (formula for tonifying the kidney essence) on skeletal development and brain tissue antioxidation in mice with kidney essence insufficiency. METHODS: Fifty ma...OBJECTIVE: To investigate the effects of Zibushenjing Fang (formula for tonifying the kidney essence) on skeletal development and brain tissue antioxidation in mice with kidney essence insufficiency. METHODS: Fifty male Kunming mice were randomly divided into five groups: normal group, model group, Jinkuishenqi Wan group, Zibushenjing Fang high dose group, and Zibushenjing Fang low dose group, with 10 mice in each group. The model of kidney essence insufficiency syndrome was established in all the mice except the normal group by using a cat to threaten the mice and by swimming until exhaustion daily which lasted about 21 days. Mice in the model group were administered 20 mL/ kg -1 · d -1 of normal saline intragastrically. The Jinkuishenqi Wan group was given 2.7 g/kg-1 · d-1 ofa solution of Jinkuishenqi Wan. The Zibushenjing Fang high dose group was given 20 g/kg -1 · d -1 and the Zibushenjing Fang low dose group was given 10 g/kg -1 · d -1 of a solution of Zibushenjing Fang.The general condition of all the groups was observed, including the quantity of food and water intake, swimming time, length of femur, and weight of the femur and musculus quadriceps femoris. The total superoxide dismutase (T-SOD), glutathione peroxi- dase (GSH-Px), and methane dicarboxylic aldehyde (MDA) activities in the brain tissues were detected. RESULTS: Zibushenjing Fang could improve the manifestation of kidney essence insufficiency, increase the quantity of food and water intake, swimming time, femur length, and femur and musculus quadriceps femoris weight. It could also regulate the activities of T-SOD, GSH-Px, and MDA in brain tissue. CONCLUSION: Zibushenjing Fang may play an important role in treating kidney essence insufficiency syndrome by promoting body development and improving brain tissue antioxidation.展开更多
Objective: To investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro. Methods: Chondrocytes from the knee joints of New Zealand wh...Objective: To investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro. Methods: Chondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3 (4, 5 dimethylthiazol 2 yl) 2, 5 diphenyl tetra zolium bromide. (MTT, Sigma, M2128). Results: Basic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50 500 ng/ml and that of HA was 10 50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone. Conclusions: bFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.展开更多
Objective To observe the therapeutic efficacy of warm needling moxibustion plus intra-articular injection of sodium hyaluronate in treating chondromalacia patellae and its effect on inflammatory factors in knee joint ...Objective To observe the therapeutic efficacy of warm needling moxibustion plus intra-articular injection of sodium hyaluronate in treating chondromalacia patellae and its effect on inflammatory factors in knee joint fluid.Methods Sixty-eight patients with chondromalacia patellae were randomized into a control group and an observation group,with 34 cases in each group.The control group was treated with intra-articular injection of sodium hyaluronate,while the observation group was given additional warm needling moxibustion treatment.Before and after treatment,the two groups were scored using Western Ontario and McMaster Universities osteoarthritis index(WOMAC)and visual analog scale(VAS),examined by magnetic resonance imaging(MRI)and determined for the levels of nuclear factor-KB(NF-kB),tumor necrosis factor(TNF)-αand interleukin(IL)-1βin knee joint fluid.Clinical efficacy was estimated after treatment.Results The effective rate was higher in the observation group than in the control group(P<0.05).After treatment,the scores of pain,stiffness and daily activities,as well as the general score of WOMAC declined significantly in both groups(all P<0.05),and were lower in the observation group than in the control group(all P<0.05);the time cost for climbing up and down one staircase and VAS score decreased markedly in both groups(all P<0.05),and were shorter or lower in the observation group than in the control group(both P<0.05);the MRI grading showed no significant change in the control group after intervention(P>0.05),while the grading in the observation group showed notable improvement(P<0.05),and was better than that in the control group(P<0.05);the levels of NF-kB and IL-1βin knee joint fluid dropped significantly in the control group after treatment(both P<0.05),while the levels of NF-kB,TNF-αand IL-1βin knee joint fluid all decreased significantly in the observation group(all P<0.05)and were lower than those in the control group(all P<0.05).Conclusion Warm needling moxibustion plus intra-articular injection of sodium hyaluronate can produce definite efficacy in treating chondromalacia patellae;it can mitigate the clinical symptoms,improve the lesion extent of chondromalacia and down-regulate the levels of NF-kB,TNF-αand IL-1βin knee joint fluid.展开更多
文摘Covalent modification of bovine testicular hyaluronidase with chondroitin sulphate led to changes in the pattern of glycation of native and modified enzyme in its reaction with neutral saccharides and N-acetylhexosamines. Thus, mono- and di-saccharides inactivated the native hyaluronidase to a greater extent than the chondroitin sulfate-modified enzyme. N-acetylhexosamine, on the opposite, inactivated the modified hyaluronidase to a greater extent than the native one. These properties made it possible to use native and modified hyaluronidase as an informative research system for in vivo measurement of the predominant type of saccharide agents in the circulation. The proposed approach was experimentally substantiated by obtained results of the study on these interactions of hyaluronidase derivatives with hyaluronan fragments and their mixture. In a model of post-ischemic perfusion of the rat limb, the effect of hyaluronidase derivatives and their components on restoration of the microcirculation were tracked using laser Doppler flowmetry. Native hyaluronidase accelerated the restoration of initial level of microcirculation, but modified enzyme was markedly inhibited by glycocalyx degradation products. N-acetylhexosamine was positioned at the reducing terminal of these products as a natural label for these glycocalyx fragments. These and other data obtained under various experimental conditions supported the participation of endothelial glycocalyx in microcirculation disturbances.
基金National Natural Science Foundation of Chinagrant number:50772037 and 50732003+1 种基金Science and Technology Program ofGuangdong Province of Chinagrant number:2008A030102008
文摘In this study,the macroporous calcium phosphate cement with oriented pore structure was prepared by freeze casting.SEM observation showed that the macropores in the porous calcium phosphate cement were interconnected aligned along the ice growth direction.The porosity of the as-prepared porous CPC was measured to be 87.6% by Archimede's principle.XRD patterns of specimens showed that poorly crystallized hydroxyapatite was the main phase present in the hydrated porous calcium phosphate cement.To improve the mechanical properties of the CPC scaffold,the 15% gelatine solution was infiltrated into the pores under vacuum and then the samples were freeze dried to form the CPC/gelatine composite scaffolds.After reinforced with gelatine,the compressive strength of CPC/gelatin composite increased to 5.12 MPa,around 50 times greater than that of the unreinforced macroporous CPC scaffold,which was only 0.1 MPa.And the toughness of the scaffold has been greatly improved via the gelatine reinforcement with a much greater fracture strain.SEM examination of the specimens indicated good bonding between the cement and gelatine.In conclusion,the calcium phosphate cement/gelatine composite with oriented pore structure prepared in this study might be a potential scaffold for bone tissue engineering.
基金This work was funded by the National Natural Science Foundation of China （No. 30801167）.
文摘Objective： To testify the effect of the stem cells derived from the widely distributed fat tissue on repairing full-thickness hyaline cartilage defects. Methods： Adipose-derived stem cells （ADSCs） were derived from adipose tissue and cultured in vitro. Twenty- seven New Zealand white rabbits were divided into three groups randomly. The cultured ADSCs mixed with calcium alginate gel were used to fill the full-thickness hyaline cartilage defects created at the patellafemoral joint, and the defects repaired with gel or without treatment served as control groups. After 4, 8 and 12 weeks, the reconstructed tissue was evaluated macroscopically and microscopically. Histological analysis and qualitative scoring were also performed to detect the outcome.Results： Full thickness hyaline cartilage defects were repaired completely with ADSCs-derived tissue. The result was better in ADSCs group than the control ones. The microstructure of reconstructed tissue with ADSCs was similar to that of hyaline cartilage and contained more cells and regular matrix fibers, being better than other groups. Plenty of collagen fibers around cells could be seen under transmission electron microscopy. Statistical analysis revealed a significant difference in comparison with other groups at each time point （t=4.360, P〈0.01）. Conclusion： These results indicate that stem cells derived from mature adipose without induction possess the ability to repair cartilage defects
基金ThisprojectwassupportedbyagrantfromtheMinistryofHealth (No .96 1 14 5 )
文摘Objective: To explore the possibility of repairi ng long segmental bone defects and preventing infection with cefazolin loaded bo ne matrix gelatin (C-BMG).Methods: C-BMG was made from putting cefazolin into BMG by vac uum absorption and lyophilization techniques. The sustaining period of effective drug concentration in vitro and in vivo was detected. The time of inhibiting ba cteria,and the drug concentration in local tissues (bone and muscle) and plasma after implantation of C-BMG were examined by high performance liquid chromatog raphy. Results: The effective inhibition time to staphylococcus aureus of C-BMG was 22 days in vitro; while 14 days in vivo. The cefazolin concentrat ion in local tissues was higher in early stage,and later it kept a stable and l ow drug release. C-BMG showed an excellent ability to repair segmental long bon e defects.Conclusions: C-BMG can gradually release cefazolin with effect ive drug concentration and has excellent ability to repair segmental bone defect s. It can be used to repair segmental long bone defects and prevent infection af ter operation.
文摘OBJECTIVE: To investigate the effects of Zibushenjing Fang (formula for tonifying the kidney essence) on skeletal development and brain tissue antioxidation in mice with kidney essence insufficiency. METHODS: Fifty male Kunming mice were randomly divided into five groups: normal group, model group, Jinkuishenqi Wan group, Zibushenjing Fang high dose group, and Zibushenjing Fang low dose group, with 10 mice in each group. The model of kidney essence insufficiency syndrome was established in all the mice except the normal group by using a cat to threaten the mice and by swimming until exhaustion daily which lasted about 21 days. Mice in the model group were administered 20 mL/ kg -1 · d -1 of normal saline intragastrically. The Jinkuishenqi Wan group was given 2.7 g/kg-1 · d-1 ofa solution of Jinkuishenqi Wan. The Zibushenjing Fang high dose group was given 20 g/kg -1 · d -1 and the Zibushenjing Fang low dose group was given 10 g/kg -1 · d -1 of a solution of Zibushenjing Fang.The general condition of all the groups was observed, including the quantity of food and water intake, swimming time, length of femur, and weight of the femur and musculus quadriceps femoris. The total superoxide dismutase (T-SOD), glutathione peroxi- dase (GSH-Px), and methane dicarboxylic aldehyde (MDA) activities in the brain tissues were detected. RESULTS: Zibushenjing Fang could improve the manifestation of kidney essence insufficiency, increase the quantity of food and water intake, swimming time, femur length, and femur and musculus quadriceps femoris weight. It could also regulate the activities of T-SOD, GSH-Px, and MDA in brain tissue. CONCLUSION: Zibushenjing Fang may play an important role in treating kidney essence insufficiency syndrome by promoting body development and improving brain tissue antioxidation.
文摘Objective: To investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro. Methods: Chondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3 (4, 5 dimethylthiazol 2 yl) 2, 5 diphenyl tetra zolium bromide. (MTT, Sigma, M2128). Results: Basic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50 500 ng/ml and that of HA was 10 50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone. Conclusions: bFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.
文摘Objective To observe the therapeutic efficacy of warm needling moxibustion plus intra-articular injection of sodium hyaluronate in treating chondromalacia patellae and its effect on inflammatory factors in knee joint fluid.Methods Sixty-eight patients with chondromalacia patellae were randomized into a control group and an observation group,with 34 cases in each group.The control group was treated with intra-articular injection of sodium hyaluronate,while the observation group was given additional warm needling moxibustion treatment.Before and after treatment,the two groups were scored using Western Ontario and McMaster Universities osteoarthritis index(WOMAC)and visual analog scale(VAS),examined by magnetic resonance imaging(MRI)and determined for the levels of nuclear factor-KB(NF-kB),tumor necrosis factor(TNF)-αand interleukin(IL)-1βin knee joint fluid.Clinical efficacy was estimated after treatment.Results The effective rate was higher in the observation group than in the control group(P<0.05).After treatment,the scores of pain,stiffness and daily activities,as well as the general score of WOMAC declined significantly in both groups(all P<0.05),and were lower in the observation group than in the control group(all P<0.05);the time cost for climbing up and down one staircase and VAS score decreased markedly in both groups(all P<0.05),and were shorter or lower in the observation group than in the control group(both P<0.05);the MRI grading showed no significant change in the control group after intervention(P>0.05),while the grading in the observation group showed notable improvement(P<0.05),and was better than that in the control group(P<0.05);the levels of NF-kB and IL-1βin knee joint fluid dropped significantly in the control group after treatment(both P<0.05),while the levels of NF-kB,TNF-αand IL-1βin knee joint fluid all decreased significantly in the observation group(all P<0.05)and were lower than those in the control group(all P<0.05).Conclusion Warm needling moxibustion plus intra-articular injection of sodium hyaluronate can produce definite efficacy in treating chondromalacia patellae;it can mitigate the clinical symptoms,improve the lesion extent of chondromalacia and down-regulate the levels of NF-kB,TNF-αand IL-1βin knee joint fluid.
