Objective To establish the anti-infective tissue engineered bones (TEBs) and evaluate the anti-infective and osteogenic effects of the infection-prevention TEBs on femoral large bone defects in goats.Methods Based on the controlledrelease antibiotic system fibrin gel-coated vancomycin alginate beads (FG-Vanco-AB),the infection-prevention TEBs were established and evaluated.They were transplanted into the critical-size defects in the right femurs of goats.TEBs without the controlled-release antibiotic system were used as controls and transplanted into the left femoral defects.The breakpoint sensitivity of vancomycin (5 mg/mL) for S.aureus was used as a standard concentration.Postoperatively,the vancomycin concentrations in the lesion site,in the adjacent site and in the circulation,as well as the anti-infective effects of the infection-prevention TEBs were evaluated by High-performance liquid chromatography (HPLC).Bone hcaling was assessed by histology,CT and ECT.The results were used to evaluate the osteogenic effect of the infection-prevention TEBs.Results Results from ESM,CLSM and in vivo tracing showed that the in vitro and in vivo survival conditions of seeded cells were analogous to those of TEBs.The effective concentration (over the bactericidal concentration) of vancomycin in bilateral defects and in blood lasted for 28 days,2 days and 7 days,respectively.The concentration of vancomycin in the femur decreased gradually from the grafted site to both ends.At 28 and 56 days postoperatively,the ECT results showed no significant difference between the right and left femurs.CT and histology demonstrated that at 14,28 and 112 days after surgery,bone defects in the bilateral femurs were repaired synchronously,and were completely covered by new bone tissue after 112 days.Conclusion The anti-infective TEBs were successfully established.FG-Vanco -AB in the transplanted sites provided the local bone tissues with anti-infective capability whilst not interfered the process of bone reconstnction and wound healing.

Objective To build the suitable index system of the clinical research program acceptance evaluation.Methods The Delphi method and the weighted average method were adopted to determine the weight of each index.Results Followed by two rounds of Delphi method, twenty five evaluation indices were selected, including three first-class indices, seven second-class indices and fifteen third-class indices, and the weight of each index was calculated.Conclusions The index system,simple and reasonable, reflects the characteristics of clinical applied researches to a certain extent with a certain degree of operability, laid the foundation for the further supporting to carry out scientific research projects.

Aim To explore the effects of mangiferin on tissue factor(TF) expression in human umbilical vein endothelial cells(HUVECs) and the underlying mechanisms.Methods HUVECs were isolated and primarily cultured in vitro.After the treatment with mangiferin and oxidized low density lipoprotein(oxLDL), TF expression was determined in HUVECs with real-time PCR and Western blot.Results oxLDLinduced the mRNA and protein expression and pro-thrombotic activity of TF in HUVECs.However, the inductive effects of oxLDL were blocked significantly by mangiferin.Furthermore, mangiferin modified TF expression and activity in a dose-dependent manner.Mangiferin was demonstrated to enhance the activity of peroxisome proliferator-activated receptor gamma(PPARγ).In contrast, GW9662, an antagonist of PPARγ, reversed at least partially the suppressive effects of mangiferin on TF.Conclusion Through activating PPARγ, mangiferin suppresses the expression of TF serving pro-thrombotic functions in endothelial cells.

Liver X receptor (LXR), a member of the superfamily of nuclear receptors, plays an important role in the activation of transcription factors involved in cholesterol metabolism, glucose homeostasis inflammation and lipogenesis. It is shown that LXR agnoists have the potentiality to be used as drugs for the prevention and treatment of atherosclerosis, which is its best investigated therapeutic indication. There are many compounds being studied in preclinical evaluation and biological assay. This paper will review briefly the LXR agonists in recent years.
ATP-Binding Cassette Transporters ; metabolism ; Amines ; chemical synthesis ; chemistry ; pharmacology ; Animals ; Atherosclerosis ; drug therapy ; metabolism ; Benzimidazoles ; chemical synthesis ; chemistry ; pharmacology ; Cholesterol ; analogs & derivatives ; pharmacology ; Glucose ; analogs & derivatives ; pharmacology ; Humans ; Lipid Metabolism ; Lipogenesis ; Liver X Receptors ; Orphan Nuclear Receptors ; agonists ; physiology ; Quinolines ; chemical synthesis ; chemistry ; pharmacology ; Sterol Regulatory Element Binding Protein 1 ; metabolism

Objective To discuss a optimal culture method of primary hippocampal neurons and a more suitable method of mor-phological observation ,and provide basis to the study of synapse in Alzheimer′s Disease .Methods Postnatal 0 -1 days (P0 -1 ) C57BL/6J mice were decollated and bilateral hippocampus were separated .Low level concentration of trypsin and mechanical disso-ciation were adopted .And culture medium without serum was used to culture neurons .After 17 days culturing ,transfected neurons with Green Fluorescent Protein(GFP) by calcium phosphate precipitation ,and then observed neurons and spines by fluorescence mi-croscope .Results The neurons looked good and healthy by using this method .And the axons ,dendrites and spines which were typ-ical structure of neurons were observed clearly after transfected with GFP .Conclusion The cultured hippocampal neurons look good by this method .And the morphological characteristics of neurons and spines are observed much more clearly after transfected GFP by calcium phosphate precipitation .

BACKGROUNDRemodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta1 (TGFbeta1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGFbeta1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.

METHODSAdenoviral vector containing TGFbeta1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGFbeta1, the expression of VEGF165 and TGFbeta1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGFbeta1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.

RESULTSThe results showed the successful construction of a recombinant co-expression adenovirus vector containing TGFbeta1 and VEGF165 genes. Co-expression of TGFbeta1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.

CONCLUSIONCo-expression of TGFbeta1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.

Adenoviridae ; genetics ; Animals ; Anterior Cruciate Ligament ; cytology ; metabolism ; Cell Movement ; Cells, Cultured ; Collagen ; genetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibroblasts ; physiology ; Fibronectins ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing

3a and 7a are nonstructural proteins of SARSCoV, which are encoded separately by ORF 3a and ORF 7a in SARSCoV genome. The expression of 3a has been founded in cells infected by virus in vivo or in vitro. Firstly, the pGL3Control vector was reconstructed , the pGL3Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN? promoter gene was cloned into the pGL3Enhancer vector and pGLIP21, the Luciferase reporter plasmid with IFN? promoter was established. The availability of pGLIP21 was verified by NDV ,the inductor of IFN?, the Luciferase activity was assayed in cells transfected with pGLIP21 by Luminometer. In order to see the function of 3a and 7a protein of SARSCoV,CHO cells expressing 3a or 7a protein were transfected with pGLIP21, the intensity of luciferase activity was analyzed . By analysis, in vitro, 3a and 7a protein of SARSCoV had the similar ability in triggering the expression of Luceferase gene, i.e 3a and 7a protein of SARSCoV could effectively activate the promoter fragment of IFN? gene. This result will help studying the function of 3a and 7a protein and provide a method to study the nosogenesis mechanism of SARSCoV.

Dyspepsia is a syndrome consisting of epigastric pain, burning, fullness, discomfort, early satiety, nausea, vomiting and belching. Functional dyspepsia (FD) is diagnosed if upper gastrointestinal endoscopy does not show structural abnormality explaining these symptoms. 8%-30% and 8%-23% of Asian people suffer from of uninvestigated dyspepsia and FD, respectively. Most patients with uninvestigated dyspepsia are found to have FD. Patients with FD are usually young and there is no predilection to any gender. Overlap of FD with other functional bowel diseases such as irritable bowel syndrome and gastroesophageal reflux disease is common in Asia. Cultural difference in reporting of symptoms of dyspepsia is well-known. Moreover, dietary factors, socio-cultural and psychological issues, gastrointestinal infection including that caused by Helicobacter pylori, frequency of organic diseases such as peptic ulcer and gastric cancer responsible for dyspeptic symptoms in the study population may also influence epidemiology of dyspepsia. There is considerable heterogeneity in the above issues among different Asian countries. More studies on epidemiology of FD are needed in Asia.
Asia ; Asian Continental Ancestry Group ; Burns ; Dyspepsia ; Endoscopy, Gastrointestinal ; Eructation ; Gastroesophageal Reflux ; Gastrointestinal Diseases ; Gastroparesis ; Helicobacter pylori ; Humans ; Irritable Bowel Syndrome ; Nausea ; Peptic Ulcer ; Population Characteristics ; Quality of Life ; Stomach Neoplasms ; Vomiting

OBJECTIVETo explore the toxic effect of fluoride on the human thyroid cells (Nthy-ori 3-1) and its mechanism.

METHODSNthy-ori 3-1 cells were exposed to 0.0, 0.1, 1.0, 3.0 mmol/L of sodium fluoride (NaF) in vitro. After 24 hours incubation, 3 (4,5-Dimethylthiazol-z-yl)-3, 5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) assay were used to measure cell viability and the LDH leakage rate. Reactive oxygen species (ROS) level, constituent ratio of the cell cycle, and apoptosis rate were measured by flow cytometry.

RESULTSComparing to viability of control group (set as 100.00%), the cell viability of the 1.0, 3.0 mmol/L fluoride-treated groups (76.64 +/- 9.13)%, (64.04 +/- 6.32)% were significantly decreased (all P values <0.01). LDH leakage rate and ROS level of the 3.0 mmol/L fluoride-treated group ((48.66 +/-7.15)%, (29993.50 +/- 1786. 86) FI) were significantly increased (all P values <0.01) compared to control group ((35.24 +/- 3.02)%, (13021.33 +/- 1067.55) FI). The G0/G1 phase cells of the 1.0 mmol/L fluoride-treated group ((40.76 +/- 5.65)%) were lower than control group (60.09 +/- 1.76)% (P < 0.01), yet the percentage of cells in S phase ((54.05 +/- 4.59)%) were higher than the control group (32.59 +/- 2.43) % (P < 0.01). Comparing to control group ((9.64 +/- 3.44)%), the percentage of apoptosis cells increased in the 3.0 mmol/L fluoride-treated group ((20.09 +/- 3.22)%) (P < 0.01).

CONCLUSIONTo Nthy-ori 3-1 cells, fluoride under experimental concentrations decreases cell viability, improve the LDH leakage rate, and ROS level. It blocks the cells in S phase and induce cell apoptosis.

Apoptosis ; drug effects ; Cell Cycle ; Cell Division ; Cell Line ; Fluorides ; toxicity ; Humans ; Reactive Oxygen Species ; analysis ; Thyroid Gland ; cytology ; drug effects

OBJECTIVETo detect the IgH-MMSET fusion gene resulted from t (4;14) translocation in multiple myeloma and illuminate its significance.

METHODSIgH-MMSET fusion gene was detected in bone marrow specimens of 25 multiple myeloma (MM) patients and MM cell line NCI-H929 using reverse-transcription PCR (RT-PCR) assay followed by nested PCR to increase the sensitivity. The purified PCR products were cloned into pGEM-T vector and then sequenced using M13 forward primers. The fragment sequences were compared with that in GenBank to find matched sequences.

RESULTSOnly a 438 base pair long fragment was obtained after RT-PCR assay and was confirmed by sequencing to be a fusion gene product of IgH gene and MMSET gene in MM cell line NCI-H929. The breakpoints were located within the C micro region of IgH gene on chromosome 14 and intron 3 of MMSET gene on chromosome 4. IgH-MMSET hybrid transcripts were detected in 3 of 25 MM patients through nested PCR assay. The amplified fragments of the 3 patients were 237 base pairs (bp), 239 bp and 239 bp in length, respectively. The breakpoints on chromosome 4 were identical to that of NCI-H929 cell.

CONCLUSIONSThe formation of IgH-MMSET fusion gene is resulted from t (4;14) translocation in MM. The incidence rate is 12.0%. The presence of IgH-MMSET fusion gene may predict poor prognosis.

Adult ; Aged ; Base Sequence ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 4 ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; Oncogene Proteins, Fusion ; genetics ; Protein-Tyrosine Kinases ; Receptor, Fibroblast Growth Factor, Type 3 ; Receptors, Fibroblast Growth Factor ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Translocation, Genetic