Objective:To observe the effect of overdose fluoride on the expression of bFGF in rat's dental pulps.Methods:NaF at 20 mg/(kg?d) was given to each of 10 Wister rats by stomach perfusion for 8 weeks, another 10 rats were given the same volume of distilled water as the controls.After treatment,HE staining was used to observe the morphology of the teeth, immunohistochemical staining was adopted for study the expression of bFGF in the dental pulps of the rats.Results:The enamel layer and dentin layer of the incisors in NaF treated rats were thinner than those in the controls,the interglobular dentin was increased and the expression of bFGF in dental pulp and inner dentin was inhibited in the NaF treated rats(P

Diabetic nephropathy is a complication of diabete and its incidence is increasing obviously these years. The pathogenic mechanisms of DN are complex, with multiple factors involved such as the glycometabolism disorders , hemodynamic changes, aldose reductase pathway activation, alterations in cytokines, oxidative stress, protein kinase C activation, lipid metabolism disorders and genetic susceptibility etc, which are not completely clear yet. Thus, the research progress of DN pathogenesis is to be reviewed.

Objective To study the effects of atorvastatin on the expressions of peroxisome proliferators-activated receptor gamma(PPAR?)and matrix metalloproteinase-9(MMP-9)in atherosclerosis(AS) tissue.Methods Atherosclerotic rabbit models on aortas were established with the high-cholesterol diet,and animals were divided into control,model and atorvastatin groups(n=8).The expressions of PPAR? and MMP-9 and the effects of atorvastatin on them were observed by means of immunohistochemistry.Results The expression of PPAR? in model group(14.38%?2.58%)was higher than that in control group(7.82%?0.96%)(P

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

Objective:To investigate the use of antibacterial agents for emergency patients with acute upper respiratory infections in tertiary hospitals in Beijing .Methods:We used the medical claim data for urban workers in 10 tertiary hospitals in Beijing from Oct .2010 to Sep.2012.Medical records of emer-gency patients with acute upper respiratory tract infections had been selected as the study sample .The proportions of antibacterial prescriptions and categories of antibacterial drugs were described and ana -lyzed.Results:This study included 135 979 visitors (male:42.7%;mean age:43.6 ±16.2 years).The average antibacterial prescription rate was 71.2% (95%CI 71.0%-71.5%), of which the single kind use was 80.0%(95%CI 79.7%-80.2%).Among acute upper respiratory tract infections , the antibacte-rial prescription rate for acute tonsillitis visits was highest (85.1%, 95%CI 84.5%-85.6%), followed by acute laryngitis and bronchitis (81.69%, 95%CI 80.4%-82.8%), acute pharyngitis (81.4%, 95%CI 77.7% -85.0%), acute sinusitis (77.0%, 95%CI 74.6% -79.4%), acute nasopharyngitis (74.3%, 95%CI 73.7%-75.0%), and common cold (67.6%, 95%CI 67.3%-67.9%).Compared with the female group, the antibacterial prescription rate for the male was higher (73.2%, 95%CI 72.8%-73.6%vs.69.7%, 95%CI 69.4%-70.0%).Compared with the <60 years age cases, the anti-bacterial prescription rate for the ≥60 years cases was higher (72.1%, 95%CI 71.8%-72.3% vs. 66 .8%, 95%CI 66 .2%-67 .5%) .In the visitors who used antibacterial drugs , the average percentage of injection use was 50.6%(95%CI 50.3%-50.9%).The top antibacterial drugs in the list of varie-ties were the second generation cephalosporins (28.4%) , followed by the third generation cephalosporins (21.7%), fluoroquinolones (21.0%) and macrolides (17.6%).Conclusion: The antibacterial pre-scription rate for acute upper respiratory tract infections in the general hospitals in Beijing is high , and the second generation cephalosporins , third generation cephalosporins , fluoroquinolones and macrolides take the lead in the total antibacterial drugs .

Objective To investigate the changes in cytotoxicity of DC-CIK cells to human multiple myeloma RPMI 8226 cells before and after treatment with oridonin. Methods Normal human peripheral blood mononuclear cells were isolated and induced to obtain DC-CIK cells. Cytotoxicity of DC-CIK cells against RPMI 8226 cells which were treated by oridonin was analyzed by LDH releasing assay. The variation for expression of NKG2D ligands on RPMI 8226 cells were measured by flow cytometry. Results DC-CIK cells were successfully induced from the peripheral blood mononuclear cells. At the same effector to target ratio, oridonin obviously enhanced the cytotocixity of DC-CIK cells against RPMI 8226 cells (P<0.01). Flow cytometry showed the expression of NKG2D ligands ULBP1 of RPMI8226 cells was most significantly increased as the cells were treated by oridonin [(9.19 ± 1.85) vs. (15.47 ± 0.67), P<0.01]. Correlation analysis indicated that cytotocixity was positively correlated with changes in ULBP1. Conclusions Oridonin can improve the cytotoxicity of DC-CIK cells against RPMI 8226 cells, which may be related with the increased expressions of NKG2D ligands on the tumor cell surface.

Objective To establish a high purity primary culture methods of human nasal epithelial cells (HNEC) in vitro and to provide a successful primary culture model for evaluation experiments of the nasal preparation .Methods Primary culture of human nasal epithelial cells were performed with enzymatic dissociation of isolated tissue and cultured in serum-free medium .HNEC were separated through magnetic field by immunomagnetic beads .We determined the purity of the separated cells by light microscopy and flow cytometry .The morphology of HNEC was observed with a scanning electron microscope .Results Under an inverted phase microscope ,the cells morphology was paving stone shaped .Under the scanning electron microscopy ,abundant microvilli and cilia differentiation were observed .Flow cytometry showed the epithelial cells accounted for 99% .Conclusion The highly purified HNEC can be directly isolated by the magnetic cell sorting system .The cell model can be used for the basic research of nasal cavity preparation .

Objective To investigate the relationship between mtND4 point mutation in sperms and asthenospermia. Methods Fifty-six asthenospermia cases and 44 control cases were collected using the WHO criterion for defining asthenospermia, the regions of mtND4 gene were amplified by using PCR of 3 pairs primers. Consequently, the point mutation, missense mutation and multiple single nucleotide polymorphisms (SNP) were analyzed by employing sequencing technology and bioinformatics tools. Results Six mutations never before identified were found. The frequency of single point mutation T10873C and T11944C in the control group were significantly higher than those in the asthenospermia group (P＜0.05). Eight cases involved T10873C or T11944C among the 10 cases in control groups with missense mutations were found. But, there were only 2 cases with such mutation in the 10 asthenospermia cases with missense mutations (P＜0.05). The previous 20 cases of missense mutations can be described as either multiple SNP group (with T10873C or T11944C) or nonmultiple SNP group. The percentage of a range and a plus b range of multiple SNP group of sperm was significantly higher than the non-multiple SNP group(P＜0.05). Conclusions mtND4 gene mutation, especially the missense mutation may induce loss of sperm motility. The mutations of T10873C and T11944C may be useful for sperm motility or counteract the influence for the sperm motility caused by these harmful mutations.

Objective To study the serum peptide fingerprint using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) technology,and find the different peaks with potential significance and establish the diagnosis model of Staphylococcus aureus and Escherichia coli bloodstream infection.Methods To establish ICR mice model of S.aureus and E.coli bloodstream infections,and collect serum samples.The serum samples were purified by weak cation exchange beads,the serum peptide fingerprint was recognized by using MALDI-TOF MS and BioExplorer software between infections group and normal control group.Results Compared with the normal control group,6 peptides were up-regulated,7 peptides downregulated and 8 peptides up-regulated first and then down-regulated in S.aureus infection group;And 5 peptides down-regulated,4 peptides down-regulated first and then up-regulated,and 8 peptides up-regulated first and then down-regulated in E.coli infection group.Conclusion MALDI-TOF MS combined with BioExplorer software may be used as a tool to study the serum peptides of S.aureus and E.coli bloodstream infection,effectively find significant peptides for establishing a diagnosis model of these two bacterial infections,and has a certain value for the diagnosis of bacterial bloodstream infection.

Objective Subcutaneous transplantation Lewis lung carcinoma model is commonly used in experimental studies.Researchers often choose different transplantation sites to create the models while little attention was paid on the effect of different inoculation sites on the formation of transplanted tumors.The aim of this study was to compare the effect of tumor cell inoculation at different sites on tumor formation in mice.Methods Lewis lung adenocarcinoma (ll2-luc-m38) cells stably expressing luciferase protein were subcutaneously injected into C57 BL/6 mice at the right armpit, right groin, or footpad, respectively.An IVIS spectrum in vivo imaging system was used to observe the tumor and metastasis formation.The survival time and mortality were recorded.H-E stained pathology was performed to examine the histological changes of the lung tissues and tumor metastesis.Results The tumor formation time was earlier in the armpit and groin groups, both with a tumor formation rate of 100%, while the tumors occurred later, with a tumor formation rate of 33% in the footpad group.The pulmonary metastasis rate was 70% in the groin group, 50% in the ampit group, and 0% in the footpad group, at the 21st day after inoculation.The footpad group had a high mortality.The tumors in the groin group and armpit group can be surgically resected, with a postoperative survival rate of 100%.Conclusions In this mouse model of subcutaneously transplanted Lewis adenocarcinoma, the groin and ampit groups have advantages such as a high tumor formation rate, good tolerance of tumor resection, low surgical mortality rate, easy to monitor, simple operation and high reproducibility.The axillary group has an even higher metastasis rate.