Objective To culture preadipocytes in vitro and to study the cell compatibility of PLGA scaffolds,collagen scarfolds and hyaluronic acid-based scaffolds and tO choose the optimal seeding method.Methods The preadipocytes from human abdominal adipose tissue were isolated and cultured in enzyme-digesting method.The generation of human preadipocytes was planted on PLGA scaffolds,collagen scaffolds and hyaluronic acid-based scaffolds.and the cell compatibility was observed by MTT method.The seeding efficiency of human preadipocytes on scaffolds.human preadipocytes were seeded to hyaluronic acid-based scaffolds by static culture and stirred culture.Results Compared compatibility of preadipocyte with three different scaffolds,there was great difference between hyaluronic acid-based scaffolds and PLGA scaffolds.Difference also existed between hyaluronic acid-based scaffolds and collagen scaffolds that were different from PLGA scaffolds.Among them,hyaluronic acid-based scaffolds was the best.Conclusion Hyaluronic acid iS a better scaffolds material for adipose tissue engineering compared with PLGA and collagen.The seeding efficiency of stirred culture is higher than static culture,which is an optimal method for cell seeding tO 3-D scaffolds.

BACKGROUND: Smooth muscle cells (SMCs) proliferation and transmigration and platelet activation cause thrombogenesis and lead to grafted vessel restenosis. Nitric oxide (NO) can inhibit the above-mentioned biological responses, but whether endothelial nitric oxide synthase (eNOS) gene transfection can inhibit the neointimal hyperplasia in graft seeded with SMCs remains uncertain.OBJECTIVE: This study was designed to further investigate the effect of eNOS gene transfection on neointimal hyperplasia in the grafts seeded with SMCs.DESIGN, TIME AND SETTING: This study, a repeated observation and measurement experiment, was performed at the Central Laboratory and Laboratory of Molecular Biology of Xi'an Jiaotong University Medical College from April 2006 to May 2007.MATERIALS: One 1-month-old New Zealand rabbit was used to acquire SMCs. Another 18 adult New Zealand rabbits were randomly divided into 3 groups (n=6).In normal control group,the vessel graft with no SMCs were transplanted; In SMC/lacZ group, the vessel grafts with SMCs transfected with lacZ were transplanted;In SMC/eNOS group,the vessel grafts with SMCs seeded with eNOS were transplanted.METHODS: Rabbit SMCs were transduced with pseudotyped retroviral vectors, Murine leukemia virus/vesicular stomatitis virus G glycoprotein, carrying genes coding for eNOS or lacZ gene. The SMCs then were seeded on the vessel grafts and implanted into the rabbit abdominal aorta using vessel bypass transplantation.MAIN OUTCOME MEASURES: Nitric oxide (NO) content in the supernatant of cells transfected with eNOS and lacZ gene was detected by citrulline method. The grafts were stained with X-gal to visualize the seeded cells: the seeded SMCs were stained blue,while eNOS were stained red. The thickness of the neointima on a graft was measured with a microscope.RESULTS: Eighteen rabbits were all included in the final analysis. NO content in the SMC/eNOS group was significantly higher than that in the normal control group (P<0.05). The SMCs transfected with lacZ gene showed blue after X-gal staining under the inverted microscope. Thirty days after implantation, there was no difference in neointimal thickness between normal grafts and grafts seeded with eNOS or lacZ transduced SMCs (P>0.05).100 days after implantation,the neointimal thickness on grafts seeded with eNOS transduced SMCs was similar to that of unseeded grafts (P>0.05 ), but was significantly thinner than that on grafts seeded with SMCs transduced with only lacZ gene (P<0.05).CONCLUSION: eNOS gene transfection inhibits nenintimal hyperplasia in the vessel graft seeded with SMCs.

Objective To obtain the change of bcl-2/bax/fas/fasL in splenic lymphoctyes with different lasting time of hypoxicischemic brain damage (HIBD). Methods The newborn rat were divided into 6 groups by the time of being HIBD model randomly, includes 1/6/12/24/48/72 hour(s) (8 for every group),and control groups were established at the same time point. The following four apoptosis related genes bcl-2/bax/fas/fasL were tested by real time PCR. Results ( 1 ) bcl-2: the mRNA expressions of HIBD groups were lower than control groups at the same time ( P<0.01 ). Eliminated the control effects, the mRNA expressions of HIBD groups were differernt by the modeling time(P ＜0.01 ). (2)bax: the mRNA expressions of HIBD groups were higher than control groups at the same time( P ＜0.01 ), and in control group the expression of 6 h was much higher than any other groups (P<0.01 ). Eliminated the control effects, the mRNA expressions of H IBD groups were different by the modeling time( P<0.01 ). (3)bcl-2/bax: the ratios of HIBD groups were lower than control groups at the same time( P ＜0.05 ), the ratios in control groups were higher than 1 ( except for 1 h); while in HIBI) groups the ratios were lower than 1; Eliminated the control effects, the ratios were different in all the groups. (4)fas: the mRNA expressions of HIBD groups were higher than control groups at the same time ( P ＜0.01 ), and both were maximum at 6 h. (5)fasL: the mRNA expressions of HIBD groups were higher than control groups in 1 h and 6 h ( P<0.01 ), while lower than control group at other time points( P<0.01 ),the expression of 24 h was the maximum of control groups and 12 h was the maximum of HIBD groups. (6)fas/fasL: the ratios of HIBD groups were higher than control groups( P ＜0.01 ) (except for 6 h), and the ratios in control groups were lower than 1 ( P<0.01 ) ( except for 6 h), and not concentrated, while in HIBD groups were higher than 1 ( except for 24 h), between 0.69 to 5.65. Conclusion Pro-apoptosis genes ( include bax/fas/fasL) were promoted by HIBD, while anti-apoptosis gene(bcl-2) was inhibited. The maximum of pro-apoptosis genes became early in HIBD. Both the pro- and anti-apoptosis genes got their maximum at 6 h and 12 h of HIBD. The apoptosis suppression was the main effects in control groups from the ratio of bcl-2/bax, which was lower than 1. The apoptosis promotion was the main effects in HIBD groups from the ratio of bcl-2/bax, which was higher than 1, especially at 12 h. Thefas/fasL effect which is the major way of lymphocytes apoptosis was strengthened in HIBD.

Objective To establish a rat model of liver metabolism profile in chronic heart failure (CHF), to explore the dynamics of liver metabolism in CHF from the point of view of metabolism, and to find the characteristic metabolites valuable for the molecular mechanism and management of CHF.Methods Twenty male Wistar rats were assigned to the CHF group to receive aortic coarctation or to the control group to receive sham surgery, and were bred for 24 weeks following surgery.The metabolic profiling of the rat liver tissues was analyzed on a metabonomics research platform. Orthogonal partial least squares-discriminant analysis ( OPLS-DA) model and principal component analysis ( PCA) model were established for liver tissues of the CHF rats, and the characteristic metabolites were finally derived by data processing with SPSS 19.0 software.Results The PAC and OPLS-DA models were established successfully.Ten characteristic metabolites with significant differences between the CHF and control groups, including lysophosphatidyl choline, lysophosphatidyl ethanolamine, oleic acid, glycocholic acid, and dehydroepiandrosterone sulfate, were screened and identified from the models.Conclusions The metabolic disorders in CHF rats are well fitted to the established metabolic profile models, and these identified characteristic metabolites may provide reference for the pathophysiological molecular mechanism and management, etc., of chronic heart failure.

OBJECTIVETo study the effect of different composition structures of total paeony glycoside (TPG) component and total phenolic acid of Ligusticum chuanxiong ( TLPA) on sodium dithionite (Na2S2O4) -induced human umbilical vein endothelial cells (HUVEC) hypoxic injury. The baseline geometric proportion was used to design different components structure. And then the best structure of components by cell injury model were optimized.

METHODA HUVEC hypoxic injury model was established by being induced of Na2S2O4. Cell viability was measured by MTI colorimetric method, intracellular superoxide dismutase (SOD) activity, malondialdehyde (MDA), lactate dehydrogenase( LDH) levels, nitric oxide (NO) contents were measured by kits. At last, Western blot analysis was used to detect the expression of two proteins, Bcl-2 and Bax.

RESULTCompared with the model group, TPG component, TLPA component at different composition structures can significantly increase SOD activity and decrease MDA, LDH, NO levels (P < 0.01, P < 0.05). Paeoniae Radix Rubra and Chuanxiong Rhizoma components can downregulate the expression of Bax protein and upregulate the expression of Bcl-2 protein. The ratio of Bcl-2 and Bax was significantly increased (P < 0 01, P < 0 05), it means that cell apoptosis was inhibited. The results indicate that among all the component composition structures, TPG and TLPA component at the proportion of 8: 2 had the best protection on hypoxic injury of endothelial cells.

CONCLUSIONTPG component and TLPA component can resist HUVEC hypoxia injury, the protective effect was the most evident under the structure of 8: 2, which may be due to the inhibition of intracellular lipid peroxidation and cell apoptosis.

Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Glycosides ; analysis ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; metabolism ; Humans ; Hydroxybenzoates ; analysis ; pharmacology ; Hypoxia ; drug therapy ; genetics ; metabolism ; physiopathology ; Malondialdehyde ; metabolism ; Oxygen ; metabolism ; Paeonia ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Rhizome ; chemistry

BACKGROUND:Stem cel is a kind of pluripotent cels with self-replication ability, which can differentiate into various cels under certain conditions. Furthermore, stem cels are rich in a variety of growth factors, which can induce the generation of vessels and nerves, and improve the blood supply of lower limbs, thereby achieving the treatment and preventions of lower limb ischemia OBJECTIVE:To summarize and compare the recent achievements in the theory and therapeutic efficacy of stem cels from different sources in the treatment of diabetic foot. METHODS:The first and second authors retrieved PubMed, Sciencedirect and Medline databases for relevant articles published from January 2000 to January 2015. The key words were “diabetic foot, pathogenesis, stem cel therapy” in English. Initialy, 186 articles were retrieved, and finaly 44 articles were included in result analysis. RESULTS AND CONCLUSION:Stem cels can be a new choice for the treatment of diabetic foot. After stem cel therapy, corresponding symptoms have been aleviated, including the generation of new blood vessels and the reshaping of the colateral vessels, the improvement of motor nerve conduction velocity and nerve reflex, the improvement of the sense of skin pain and temperature, and pain relief. It is stil unclear whether alogeneic stem cels are safe or not, but autologous stem cels, especialy bone marrow mesenchymal stem cels, can be better able to repair damaged vessels and nerves and restore the microcirculation of blood supply. Currently, we need to do more basic and clinical researches to solve the folowing problems: to confirm the effectiveness and safety of stem cel therapy for diabetic foot; to identify whether there is a difference in the differentiation and secretory activity between stem cels in diabetic patients and ordinary people; to give ful play to the treatment of diabetic foot.

Objective To clarify the inhibitory effect of low molecular weight chitosan(LMCTS)on hepatic fibrosis induced by carbon tetrachloride (CCl4 )in the rats, and to investigate its effect on the expression of Toll-like receptor-4(TLR4 )and to lay the foundation for the development of the clinical candidate drug of liver fibrosis. Methods 72 male Wistar rats were randomly divided into blank control group, CCl4 group (model group), glycyrrhizinate (DG)group,50,100 and 150 mg·kg-1 LMCTS groups (low,middle and high doses of LMCTS groups).In addition to blank control group,the rats in the remaining groups were given 40% CCl4-vegetable oil (1.75 mL·kg-1 ),2 times per week for 8 weeks,by intraperitoneal injection to establish the model of rat hepatic fibrosis.And the rats in blank control group were injected with the same amount of 100% vegetable oil agent. From the ninth week, the rats in DG and LMCTS groups were given DG and LMCTS by intragastric administration, 1 time/week for 4 weeks. Then all rats were sacrificed, the activities of serum glutamic acid aminotransferase (ALT),aspartate aminotransferase (AST)and alkaline phosphatase (ALP)were detected with ELISA kit;the pathological changes in liver tissue were observed under light microscope, and the TLR4 expressions were detected by RT-PCR and Western blotting method. Results The serum ALT, AST and ALP activities in middle and high doses of LMCTS groups were lower than those in model group (P<0.05).The serum ALT activity in middle dose of LMCTS group was lower than that in low dose of LMCTS group (P<0.05),but the activities of AST and ALP had no statistically significant change(P>0.05).There were statistically significant differences in the serum ALT, AST and ALP activities between high dose of LMCTS group and middle dose of LMCTS group (P<0.05).There were obvious hepatocyte steatosis,inflammatory cell infiltration,collagen fiber hyperplasia and hepatic lobule damage in the rats in model group.However,all the changes in liver tissue of the rats in LMCTS group were significantly reduced, especially in high dose group. The results of RT-PCR and Western blotting method showed that the expression of TLR4 was declined in LMCTS groups compared with model group (P<0.05,P<0.01),especially in high dose of LMCTS group,and there were statistically significant differences between different doses of LMCTS groups (P<0.05).Conclusion High dose of LMCTS can decrease the serum transaminase activity of liver fibrosis rats and improve liver function,and this process may be related to declining the expression of TLR4 .

To compare the clinical efficacy of treating Denis type B thoracolumbar burst fracture with allogenic bone and calcium sulfate implanting in injured vertebra.A total of 46 patients were randomly assigned into 2 groups.Group A( n=22) received allogenic bone implanting in injured vertebra while group B( n=21) had calcium sulfate grafting in injured vertebra.Group A was better than group B in maintaining tanterior vertebral body height and lessening the degree of bone defect ( P <0.05 ).No significant differences existed in operaive duration , blood loss volume, correcting Cobb′s angle, preventing degeneration of adjacent segments , visual analogue scale ( VAS) and Japanese Orthopaedic Association ( JOA) scores and the degree of bone defect ( P>0.05).

The effect of triptolide on proliferation and apoptosis of human multiple myeloma RPMI-8226 cells in vitro, as well as the roles of nuclear factor-kappa B (NF-κB) and IκBα was investigated. The effect of tritptolide on the growth of RPMI-8226 cells was studied by MTT assay. Apoptosis was detected by Hoechest 33258 staining and Annexin V/PI double staining assay. The expression of NF-κB and IκBα was observed by Western blot and confocal microscopy. The results showed that triptolide inactivated NF-κB apoptotic pathway in human multiple myeloma RPMI-8226 cells. Triptolide at nM range induced proliferation inhibition in a dose- and time-dependent manner and apoptosis in a dose-dependent fashion in RPMI-8226 cells. Besides, we observed the inhibition of NF-κB /p65 in the nuclear fraction was correlated with the increase in the protein expression of IκBα in the cytosol. These results suggested that triptolide might exhibit its strong anti-tumor effects via inactivation of NF-κB/p65 and IκBα.

Hypecoi Erecti Herba is one of ethnic and folk medicines in China, mostly known in Tibetan medicine. It has the function of heat-clearing and detoxifying, and analgesia, indicated for the treatment of a variety of heat syndrome, such as "Chi Ba" pestilence, and warm toxin. Previous phytochemical studies revealed isoquinoline alkaloids are its main chemical components, and contemporary pharmacological research demonstrated that Hypecoi Erecti Herba has antibacterial, liver protection, analgesic, anti-inflammatory, and antiviral activities. The present paper describes the resource distribution, chemical constituents, and pharmacological activities of the plants in Hypecoum L. to provide the references for further research and development of the plants in Hypecoum L.