BACKGROUND:Based on the original advantages of silk fibroin, positive charged water-soluble chitosan modified silk fibroin is modified on surface and could improve celladhesion on the scaffolds. OBJECTIVE:To verify the biocompatibility of chitosan-modified silk fibroin with human adipose-derived stem cells (hADSCs), and feasibility of constructing tissue engineered adipose in vitro. METHODS:The hADSCs at passage 3 were seeded on chitosan-modified silk fibroin at the concentration of 1×107/L, as the experiment group;at the same cellconcentration, hADSCs were seeded in 96-wel plates as the control group. MTT tests were performed to evaluate the adhesion, growth and proliferation of hADSCs on chitosan-modified silk fibroin. Then hADSCs were implanted on the chitosan-modified silk fibroin scaffolds at the concentration of 1×109/L. The hADSCs seeded onto chitosan-modified silk fibroin complexes were respectively cultured with adipogenic differentiation medium and ordinary high-glucose DMEM. The complexes were stained with oil red O, and detected with RT-PCR after cultured 14 days. RESULTS AND CONCLUSION:The hADSCs adhered to and proliferated on the scaffolds. After cultured with adipogenic differentiation medium for 14 days, oil red O staining demonstrated that there were amount of mature adipocytes on the scaffold. The peroxisome proliferator activated receptorγ2 was positively expressed. The chitosan-modified silk fibroin possessed excellent biocompatibility in vitro. The co-cultured hADSCs could be induced to mature adipocytes successful y.

BACKGROUND:Low toxicity of Genipin has certain species and cellspecificity. Biocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels is essential for construction of
tissue-engineered adipose.
OBJECTIVE:To investigate the bbiocompatibility of Genipin cross-linked type I colagen with human adipose-derived stem cels.
METHODS:Human adipose-derived stem cels were isolated and cultured to the third generation, and the cels were seeded on Genipin cross-linked type I colagen scaffold. MTT assay was used to evaluate the adhesion and proliferation of cels on the scaffold, and the toxic effects of Genipin cross-linked type I colagen on human
adipose-derived stem cels. Optical microscopy and scanning electron microscopy were utilized to observe the adhesion and growth process of human adipose-derived stem cels on the scaffold as wel as the morphological changes of cels.
RESULTS AND CONCLUSION:Human adipose-derived stem cels could adhere to the scaffold immediately
after seeded and increase gradualy on the scaffold, with the average adhesion rate of 86.5%. Optical microscopy and scanning electron microscopy showed that human adipose-derived stem cels adhered wel on the scaffold. The cels increased gradualy over time, and could migrate into the scaffold, and distribute evenly with the passage of time when observed with optical microscopy. The result showed Genipin possesses very low cytotoxicity to the cels, and the outstanding biocompatibility is found between the cels and scaffoldin vitro after cross-linked with Genipin.

Objective To investigate the influence of IL-8 on the tight junction of vascular endothelial cells.Methods Immunofluorescence was used to observe the modality and the distribution of three tight junction proteins (occludin,claudin-5 and ZO-1) of the EA.hy926 cells treated with IL-8 under different concentrations and different times.RT-PCR was used to measure the mRNA expression of these three proteins.Results The results demonstrated that IL-8 could change the distribution of occludin,claudin-5 and ZO-1 in EA.hy926 cells,and the mRNA expression of occludin,claudin-5 and ZO-1 decreased with the increase of IL-8 concentration and treated time.Conclusion The effects of IL-8 on the distribution and the expression of occludin,claudin-5 and ZO-1 are dose and time-dependent.

BACKGROUND: Low frequency-pulsed electromagnetic fields (LF-PEMFs) have been confirmed to not only treat delayed bone union or nonunion, but also be effective for fresh bone fractures, ostectomy and fatigue fractures.OBJECTIVE: To systematically evaluate the efficacy and safety of LF-PEMFs for bone healing.METHODS: CNKI and PubMed databases were retrieved for the randomized controlled trials concerning LF-PEMFs promoting bone healing published from January 1985 to May 2016. Meta-analysis was conducted on RevMan 5.1 software after data management and quality assessment.RESULTS AND CONCLUSION: Thirteen eligible literatures were enrolled, involving 1244 patients. Meta-analysis showed that the overall effectiveness and bone mineral density in the LF-PEMFs group were significantly higher than those in the control group. The healing time in the LF-PEMFs group was significantly shorter than that in the control group. These results suggest that LF-PEMFs show high efficacy and safety for bone healing, and also can markedly shorten the healing time. However, more high-quality multi-center, large-scale randomized controlled trials are still needed to testify all above conclusions.

TGF-beta, as an inhibitor of hemopoiesis, excreted by hematopoietic stem and progenitor cells, down-regulates the expression of cytokines such as Flt-3 ligand, SCF, IL-3 etc on the stem and progenitor cells. The effect of anti-TGF-beta antibody on ex vivo expansion and expression of adhesive molecules on cord blood CD34(+) cells was studied in this research. The CD34(+) cells from six units of fresh umbilical cord blood were enriched by density gradient sedimentation and purified by miniMACS cell isolation system, and plated them into the SFEM serum free culture system which containing SCF, Flt-3L, TPO and IL-3 in the condition of 37 degrees C, 5% CO2, and saturated moisture. There were three groups in this experiment: (1) blank group: same as the culture system described above; (2) control group: added with normal rabbit IgG into the mentioned culture system; (3) test group: the same culture system with anti-TGF-beta1 antibo-dy. Cultured for 6 days, the number of mononuclear cells (MNC) was counted, the expression of CD34 antigen, CD117 (c-kit) antigen, CD11a antigen, CD49d antigen and CD33 antigen was tested with FCM. Meanwhile, cells of the three groups were plated in the methylcellulose culture system for 14 days, the number of CFU-GEMM, BFU-E, CFU-GM was counted. The results indicated that the expansion multiples of MNC, CD34(+) cells, CD34(+)c-kit(+) cells, CFU-GEMM in the test group (41.82 +/- 13.49, 15.62 +/- 6.95, 13.36 +/- 6.12, 11.07 +/- 4.05) were significantly higher than in the control group (28.86 +/- 9.03, 10.40 +/- 4.98, 9.04 +/- 4.40, 6.36 +/- 2.37) (P = 0.001, 0.002, 0.003, 0.002) respectively. The expansion multiple of more primitive CD34(+)c-kit(-) subpopulation in the test group (69.10 +/- 41.06) was even higher than in the control group (27.29 +/- 10.40) (P = 0.024). Adhesion molecule expression on the CD34(+) cells after short-term expansion: the expression of CD11a on the CD34(+) cells of the original cord blood was (61.73 +/- 4.13)%, and CD49d was (55.12 +/- 5.22)%. After expansion in each group the expression of CD11a on the CD34(+) cells did not change with statistical significance (P > 0.05), the expression of CD49d increased (P < 0.05). Compared with blank group and control group, anti-TGF-beta antibody did not impact on the expression of CD11a and CD49d (P > 0.05). It is concluded that anti-TGF-beta antibody can synergize other cytokines to effectively enhance the proliferation of cord blood NC, CD34(+) cells, progenitor subpopulation of CD34(+)c-kit(-) cells, and increase the output of more primitive progenitor colony, CFU-GEMM and BFU-E. At the same time, anti-TGF-beta antibody did not depresss the expression of adhesion molecules on CD34(+) cells.
Antibodies ; pharmacology ; Antigens, CD34 ; analysis ; CD11a Antigen ; analysis ; Cell Adhesion Molecules ; analysis ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Humans ; Integrin alpha4 ; analysis ; Pregnancy ; Proto-Oncogene Proteins c-kit ; analysis ; Transforming Growth Factor beta ; immunology

OBJECTIVETo observe the effect of the crude extracts of Dan-shen root and San-qi of different proportion on platelet aggregation and adhesion in normal rabbits.

METHODWith rabbits, ig. (4d, exsanguinated via carotid artery, percentage of platelet aggregation and adhesion was measured.

RESULT AND CONCLUSIONAspirin (4.4 mg/Kg) could markedly inhibit platelet aggregation and adhesion in normal rabbit. The proportions of Dan-shen root/San-qi (10:0, 10:1, 10:3, 10:6, 1:10) could markedly inhibit platelet aggregation, among which 10:3 was the best. San-qi alone had little effect on aggregation. The proportions of Dan-shen root/San-qi (10:3, 10:6, 0:10) could markedly inhibit platelet adhesion, among which 0:10 was the best, and the proportions(10:0, 10:1, 1:10) had little effect.

Animals ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Male ; Panax ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Platelet Adhesiveness ; drug effects ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rabbits ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).

METHODSWAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.

RESULTSWAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).

CONCLUSIONSThe WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.

Adolescent ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Proliferation ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Female ; Humans ; Infant ; Jurkat Cells ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; etiology ; metabolism ; RNA, Messenger ; analysis ; Wiskott-Aldrich Syndrome Protein Family ; analysis ; genetics ; physiology

OBJECTIVETo investigate the effect of two methods of pigmentation on the flexural strength of dental Y-TZP/porcelain layered structure.

METHODSKaVo zirconia substructures were pigmented by dipping presintered blocks in the coloring solution VITA LL1 and LL5, and colored TZ-3YS zirconia substructures were fabricated by adding pigments before isostatic pressing. The colors No.1 and No.5 were used for the test. The specimens were made in monolithic or bilayered forms, and the flexural strength was tested. XRD and SEM with EDX were used to analyze the characteristics of the surface structure.

RESULTSIn KaVo group, no significant differences were found in the flexural strength between white and LL1 and LL5 colored monoclinic materials, nor in bilayered structures. While in TZ-3YS group, significant differences were noted in the flexural strength between color No.5 white and color No.1 monoclinic materials, but not between the latter two subgroups. The flexural strength was significantly lowered by veneering with porcelain in both zirconia groups, and similar findings were observed with the monoclinic materials. Only the tetragonal phase was detected in both of the zirconia groups.

CONCLUSIONPigmentation has no apparent effects on the bonding strength between the veneering porcelain and zirconia. Both coloring methods are appropriate when the concentration of the pigments is under deliberate control.

Dental Bonding ; Dental Materials ; Dental Porcelain ; chemistry ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Pigmentation ; Tensile Strength ; Yttrium ; chemistry ; Zirconium ; chemistry

OBJECTIVETo compare the actions of the three flavone ingredients in choerospondias axillaris on arrhythmias Induced by aconitine.

METHODLangendorff perfuse was applied in the experiment, the antiarrhythmic action was to study by using aconitine on the the isolated heart; The antiarrhythmic action of the three flavone ingredients in choerospondias axillaris was to study by using i.v. aconitine in rat to induce arrhythmias.

RESULTCompared with the NS group, sample 1 and sample 2 both significantly prolonged the beginning time of VF of isolated heart and increased the dosage of aconitine, sample 3 reduced the beginning time of VF of isolated heart and decreased the dosage of aconitine, sample 1 and sample 2 both greatly prolonged the beginning time of VE, VT, VF, HA; sample 3 greatly reduced the beginning time of VT,VF. The actions of the three samples were in a concentration-dependent way.

CONCLUSIONSample 1 and sample 2 both resisted the occurrence of arrhythmias induced by aconitine, sample 3 markedly promoted the occurrence of arrhythmias induced by aconitine.

Aconitine ; Anacardiaceae ; chemistry ; Animals ; Anti-Arrhythmia Agents ; isolation & purification ; therapeutic use ; Arrhythmias, Cardiac ; chemically induced ; prevention & control ; Dose-Response Relationship, Drug ; Female ; Flavones ; isolation & purification ; therapeutic use ; In Vitro Techniques ; Male ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar

The possibility of using a subunit or fragment of human chorionic gonadotropin (hCG) as an immunogen for birth control has been actively explored for many years. This protein homone is produced by the fertilized egg and is required for implantation of the blastocyst into the maternal uterus and the maitenance of pregnancy. In previous studies, several bio-synthesized hCG chimeric peptides (CP) that contain three linear B-cell epitopes (beta5, beta9 and beta8) of beta-hCG subunit together with various foreign 'promiscuous' T-cell epitopes were constructed and expressed as potential new hCG vaccine immunogens. In order to detect antibodies to each of the individual B-cell epitopes present in the animal antiserum raised against the hCG CPs, we decided to construct three recombinant proteins, each contains a single target B-cell epitope (betaE) of beta-hCG. Two sets of DNA fragments were chemically synthesized encoding the beta5, beta9 and beta8 epitopes (betaE) 45 approximately 52, 113 approximately 116 or 133 approximately 144 of beta-hCG subunit and were inserted into the downstream of streptavidin (Stv) gene in pTSA18 separately, with or without an extra TAA codon at the 3'-terminals of the genes. SDS-PAGE analysis revealed that only Stv-betaE (-beta5, -beta9 or -beta8) fusion genes set with the TAA codon can be expressed in E. coli BL21 (DE3) pLysS strain at high level after 1mM IPTG induction for 4 hours. Additionally, these fusion proteins can all be recognized by specific polyclonal antiserum (RS-4157) generated upon immunization with the loop peptide 38 approximately 57 of beta-hCG, monoclonal antibody (mAb) FB12 to beta9 epitope and mAb OT3A that specially recognizes reporter sequence 133 approximately 139 of beta8 epitope 137 approximately 144. Each of the proteins can be purified to 95% relative homogeneity using an improved method of preparative gel polyacrylamide gel electrophoresis. The yields were 5 mg per 1 L culture. The three target Stv-betaE fusion proteins will be useful in determining the immunogenicity of designed hCG CPs and hCG vaccines, including hCG DNA vaccines.
Chorionic Gonadotropin, beta Subunit, Human ; genetics ; immunology ; Epitopes, B-Lymphocyte ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; isolation & purification ; Streptavidin ; genetics ; Vaccines, Synthetic ; immunology