BACKGROUND:The auditory ossicle chain reconstruction is stil an important method to treat conductive deafness. Although a great variety of materials have been applied, the blood supply of otosteon after the implantation is ignored. Moreover, there is no real bone formed.
OBJECTIVE:To observe the angiogenesis of vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold which is implanted into the otocyst of guinea pig.
METHODS:Total y 60 guinea pigs were randomly divided into experimental group (vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold), col agen I control group (col agen I modifiedβ-tricalcium phosphate porous scaffold) and blank control group (β-tricalcium phosphate porous scaffold). The guinea pigs were executed under anesthesia at weeks 1, 2, 3, 4 respectively. The surface of scaffolds was observed by scanning electron microscopy. The angiogenesis of scaffolds were observed by hematoxylin-eosin staining and CD34 immunohistochemistry staining, and then the microvascular density was counted. The osteogenesis of the scaffolds was observed by toluidine blue staining.
RESULTS AND CONCLUSION:Endothelial cel proliferation and lumen formation could be observed after 1 week in the experimental group, and the angiogenesis reach the peak after 3 weeks with traffic branches formedbetween micropores. In the other two groups, the lumen formed at 2 weeks but no traffic branches were visible. The sprouting of new blood vessels in the pores were observed more in the experimental group than the other two groups (P<0.05). The adherence and proliferation of cel s could be examined in the surface and pores of the scaffold by scanning electron microscope. After 4 weeks, the osteogenesis could be observed by toluidine blue staining, especial y in the experimental group. These findings suggest that the vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold can realize an effective vascularization in the environment of guinea pigs’ middle ear. What’s more, the scaffold also can promote bone formation.

The genetic analysis of herpesviruses has been a constant challenge, due to the large, complex genomes of herpesviruses and mutagenesis of viral genes by conventional recombination methods in cell culture. Recently, a completely new approach for full-length infectious clones of herpesviruses based on bacterial artificial chromosomes (BACs) has been developed. This technique allows the maintenance, propagation and genetic modification of the viral genome as a BAC plasmid in E.coli, thus making the procedures fast, safe and effective in prokaryotic cells. This technique also makes it possible for the reconstitution of viral progeny or mutants by transfection of the BAC plasmid into eukaryotic cells, thereby facilitating the analysis of viral gene functions in the context of genome. In this presentation, Epstein-Barr virus was used as an example to describe the principle, establishment of the technique and mutation introduction into the BAC plasmid, and to discuss the perspective in the use of BAC-cloned herpesviruses.

25 is also an important impact factor among various components.

OBJECTIVETo explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

Bidens ; chemistry ; Child ; Flavones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Immunoglobulin A ; blood ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Purpura, Schoenlein-Henoch ; blood ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Animals ; Calcitonin Gene-Related Peptide ; pharmacology ; Ischemic Postconditioning ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley

The Rhei Radix et Rhizoma was one of the most widely used traditional Chinese medicine for its special biological activities. The content of rhein, one of its major compounds, was an important standard for the quantity control of Rhei Radix et Rhizoma. The major method used for the detection of rhein was instrumental analysis like HPLC, but it was complex, time-consuming and cannot detect large samples at the same time. The enzyme-linked imunmosorbent assay (ELISA) was accurate, reliable, simple, low costs, and of a high-throughout. Recently, it was widely used for the determination of those small molecule compounds in some traditional Chinese medicinal plants. In this study, an artificial antigen were synthesized by the carbodiimide (CDI) method. Rhein-bovine (rhein-BSA) conju gate and rhein-ovalbumin (rhein-OVA) conjugate, were produced as the immunogen and coating antigen, respectively. The conjugate and the hapten number in the conjugate were determined by UV-Vis spectrophotometry (UV). The conjugation ratio of Rhein and BSA was about 4.0:1, rhein acid and OVA was 2.6 : 1, respectively. Rhein-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The antiserum titer of the Rhein were higher than 8000 detected by ELISA. The successfully synthesized conjugate antigen rhein-BSA implies its feasibility in the establishment of fast immunoassay for the rhein content determination.
Animals ; Anthraquinones ; analysis ; immunology ; Antibodies ; analysis ; immunology ; Antigens, Plant ; analysis ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Mice ; Mice, Inbred BALB C ; Rheum ; chemistry ; immunology ; Rhizome ; chemistry ; immunology

Objective Using the third high sensitive cardiac troponin I assay to establish the reference intervals of serum cardiac marker high sensitivity cardiac troponin I ( hs-cTnI ) in apparently healthy subjects of Beijing area, and to explore the relationship between cTnI and other parameters.Methods Using random selection method, a total of 440 serum samples were collected from apparently healthy subjects of Beijing area.Among them, 217 cases were male, average age was (50.0 ±14.3) years old;223 cases were female, average age was ( 49.0 ±14.0 ) years old.All of these serum samples were detected hs-cTnI using i2000 Abott biochemical analyzer and ARCHITECT STAT Abbott assay kit.And the 99th percentile upper reference limit was used as the reference interval.Non-parametric tests were used to compare the different groups, the U Mann-Whitney test was used between two groups, the Kruskal-Wallis test was used in many groups, and the correlation analysis was used Spearman method.Results The 99th percentiles of hs-cTnI in 440 apparently healthy subjects is 9.66 ng/L.While the male group is 9.66 ng/L, and the female group is 8.25 ng/L.The serum hs-cTnI level of male is higher than female ( 9.95 ng/L vs 8.25 ng/L, P<0.05 ).According to the spearman correlation analysis, the serum hs-cTnI level appears positive correlation with age, brain natriuretic peptide (BNP), alanine aminotransferase (ALT),aspartate transaminase ( AST) ,gamma-glutamyl transferase ( GGT) ,total bilirubin ( TBIL) ,serum creatinine ( SCR) , hemoglobin (HGB)(P<0.05,R>0),and appears negative correlation with platelet (PLT)(P<0.05,R<0).Conclusion The serum hs-cTnI level apparently differents with gender, so it is necessary to establish reference intervals among different gender group of Beijing area.

Cleistocalycis Operculati Cortex is the dry bark of Cleistocalyx operculatus. It is the raw material of Compound Hibiscuse which is external sterilization antipruritic drugs. The quality standard of Cleistocalycis Operculati Cortex in Guangdong Province "standard for the traditional Chinese medicine" (second volumes) only contains TLC identification. It is unable to effectively monitor and control the quality of Cleistocalycis Operculati Cortex. A reversed-phase HPLC method was established for the determination of 3, 3'-O-dimethylellagic acid from Cleistocalycis Operculati Cortex and the content was calculated by external standard method for the first time. Under the selected chromatographic conditions, the target components between peaks to achieve effective separation. 3,3'-O- dimethylellagic acid standard solution at the concentration of 1.00 - 25.0 mg x L(-1) showed a good linear relationship. The standard curve was Y = 77.33X + 7.904, r = 0.999 5. The average recovery was 101.0%, RSD was 1.3%. The HPLC method for the determination of 3,3'-O-dimethylellagic acid in Cleistocalycis Operculati Cortex is accurate and reliable. It can provide a strong technical support for monitoring the quality of Cleistocalycis Operculati Cortex.
Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Bark ; chemistry ; Syzygium ; chemistry

OBJECTIVETo study the endothelioid differentiation effect of Epimedin C on murine embryonic mesenchymal stem cells (C3H/10T1/2).

METHODSC3H/10T1/2 cells were cultivated in vitro. The cytotoxicity of Epimedin C at different concentrations was determined by MTT assay and crystal violet assay. Morphological changes were observed under microscope after treated with Epimendin C. The effect of Epimendin C on the cell cycle distribution was determined by flow cytometry. mRNA expression levels of endothelial markers, such as CD31, CD34, vascular endothelial zinc finger 1 (Vezf1), angiopoietin 1 (Ang1), and angiopoietin 2 (Ang2) were detected by semi-quantitative PCR. Protein expression levels of platelet endothelial adhesive molecule 1 (CD31), ecto-5'-nucleotidase (CD73), endothelial cell specific molecule-1 (ESM-1), and integrin β5 were determined by immunocytochemical (IHC) staining.

RESULTSEpimedin C could not affect the survival rate of C3H/10T1/2 cells at 1-30 μmol/L. Its cell cycle distribution was not significantly changed after treated by 30 μmol/L Epimedin C for 24 h. C3H/10T1/2 cells were differentiated to vascular endothelial cells by Epimedin C treatment, with significant morphological changes (whirlpool-like structure). PCR results indicated that mRNA levels of classic endothelial mark- ers, namely CD34, Vezf1, Ang1, and Ang2 were significantly increased in C3H/10T1/2 cells after treated with Epimedin C for 5 days (P < 0.05, P < 0.01). Protein expression levels of CD31, CD73, and ESM-1 were also positively expressed after treated with Epimedin C for 5 days, showing statistical difference when compared with those of the control group (P < 0.01, P < 0.05).

CONCLUSIONEpimendin C could induce C3H/10T1/2 cells to differentiate into endothelioid cells.

Animals ; Cell Differentiation ; drug effects ; Cell Line ; Cells, Cultured ; Flavonoids ; pharmacology ; therapeutic use ; In Vitro Techniques ; Mesenchymal Stromal Cells ; physiology ; Mice ; RNA, Messenger