Objective To understand the role of peroxisomes on the intracellular survival of Salmonella typhimurium in macrophages. Methods Vesicles from infected or uninfected RAW264.7 macrophage-like cells were isolated by stepwise centrifugation after cells were broken by nitrogen cavitation and purified with magnetic beads containing polyclonal antibodies to TassC (Target for Salmonella secreted protein SpiC). Western-blot was used to detect the characters of the binding vesicles. Three-dimensional (xyz) fluorescence microscopy was used to determine the recruitment of peroxisomes tagged with pDsRed2-Perxi to the SCV after infection of RAW264.7 cells with Salmonella typhimurium mutant strains spiC:kan producing GFP. Volocity software was employed for image analysis. Overlap of individual fluorescence pixels from separated channels for each optical plane was determined with the Volocity 4.1 colocalization module. Results It was shown by western-blot that the anti-TassC magnetic beads could bind TassC and peroxisomal marker catalase in infected or uninfected RAW264.7 cells. Inducible nitric oxide synthase (iNOS) could be detected in infected samples, but couldn’t in uninfected samples. Lysosome associated membrane protein (LAMP1), one of the lysosome markers, and a bacterium marker RecA could not be detected from the elution of anti-TassC magnetic beads. It was determined by a fluorescence microscopy that the recruitment or overlapping of peroxisomes tagged with pDsRed2-Perxi to the Salmonella-containing vacuoles after infection of RAW264.7 cells with Salmonella mutant strains producing GFP. Calculation by Volocity 4.1 showed that peroxisome abundance increased by 1.2- or 1.3-fold, respectively, at 1h and 24h time point of infection in the infected macrophages than in uninfected cells while the bacteria abundance decreased with the infection time. Conclusion It is suggested that TassC is localized with peroxisomes, but not with lysosomes. Inducible nitric oxide synthase might be localized to peroxisomes and recruited to the SCV during infection with Salmonella mutant strains for killing the bacterium.

Objective:To establish the quality standard for Baochunchangshouling Capsules. Methods: Tanshinone Ⅱ A of Baochunchangshouling Capsules was determined by dual wavelength TLC scanning, Raxdix Glycyrrhizae, Rhizoma Zingiberis were identified by TLC. Polyrhachis Vicina Roger, Pheretima, Concha ostreae were distinguished by Microscopic identification. Results: The average recovery was up to 98.21% and RSD was 0.10% . TLC spot develops were fairly clear, and the blank test showed no interference. Conclusion: This method is reliable. The results are stable with good reproducibility. This method can be used for quality control of the capsules.

Objective To obtain recombinant neural cell adhension molecule L1(L1) and to study its structure and biological activity.Methods The cDNA encoding the mature rat L1 was isolated using RT-PCR from total RNA extracted from newborn SD-rat hippocampus tissue.The expression plasmid pETL1 mutants of several extracellular domains of(L1) were constructed by inserting L1 and its mutants cDNA into plasmid pET-28a(+) containing T7 promoter and transformed into E.coli BL21(DE3). A series expression strain BLL1 mutants were selected.Recombinant L1 and its mutant proteins were expressed at levels about 18.5%～30% of total bacteria protein in form of inclusion body after the induction.By Ni~(2+) chelation affinity chromatography,up to 90% L1's mutant proteins were purified.The expressed plasmid pcDNA3-L1 were transfected into PC12 cells and constructed PC12-engineered cells which stably and highly expressed the L1.Results Purified and refolded IgI-FN5、Ig(Ⅰ-Ⅵ) and FN(1-5) fragments could significantly promote the neurite outgrowth of PC12-engineered cells;fragment Ig(Ⅴ-Ⅵ) also can promote the neurite outgrowth but not soobvious as the before;fragment FN(3-5) have no function to induce the neurite outgrowth of PC12-engineered cells.Conclusions These results suggested that there are at least two segments in extracellular domains of L1 Ig(Ⅰ-Ⅵ) and FN(1-5) fragments are critical for inducing the neurite outgrowth of PC12-L1 cells and the key amino acids for signaling transduction located in the segments of Ig(Ⅰ-Ⅵ) and FN(1-5);fragment Ig(Ⅴ-Ⅵ) is necessary but not crucial for promoting the neurite outgrowth;fragment FN(3-5) is not important for L1 biological function.

Objective To evaluate the iron metabolism in host-pathogen interplay.MethodsA total of 13 expression profiles of iron metabolism related genes of RAW264.7 murine macrophages uninfected or infected with Salmonella typhimurium were stuldied by real-time PCR.Results The live wild-type Salmonella typhimurium induced the expression of the transferrin receptor(Tfr1)in host cell macrophages,which resulted in a sustained increase of the labile iron pool inside the host oell 1h or 24h after infection.Gene expression analysis showed that wild-type Salmonella typhimurium drove an active iron acquisition program with induction of ferrireductase(Steap3),iron membrane transporter Dmtl,and iron regulatory proteins(Irpl and Irp2),and a little of iron efflux change through ferriportin(Fpnl).On the other hand,the Salmonella mutant strain spiC-used in present studies did not cause an increase mRNA level for Tfrl at 1h or 24h,but led to an increase mRNA level for Epnl at 24h as compared with 1h. The assessment was that the labile iron pool decreased after infection with spiC-Salmonella for 24h.Conclusion Wild-type Salmonella typhimurium appears to drive an active transferrin-mediated iron uptake program after infection 1h or 24h.The spiC-Salmonella mutant strain used in our studies shows an increased iron efflux from infected cells at 24h as comparde with 1h.

This paper discusses practice of training graduate students’communication and mentoring ability.The mentors help the graduate students communicate with the undergraduates to grasp the experiment skills through evaluation on the spot. Communication and discussion skills are used to transmit the graduate students’ standards and expectation to help them get the correct contents.The strategies help undergraduates become outstanding experimentalists and improve quality of graduate training.

Head ; surgery ; Humans ; Neck ; surgery ; Reconstructive Surgical Procedures

OBJECTIVE　To establish a quality standard for Tiaogu tablet.METHODS　TLC was used to make a qualitative determination for 4 drugs in the formulary.The content of bomeol was determined by GC, the contents of strychnine and brucine were determined by TLC.RESULTS　4 drugs in the formulary were determined under stable condition with high sensitivity and precise results.CONCLUSION　This study may be helpful to develop the quality standard for Tiaogu tablet.

On the basis of the theories of air sterilization, the Virobuster steritubes are very effective in the removal and inactivation of airborne bacteria species, viral particles, fungi and spore by using the high-intensity UV lamp and close-in UVC treatment room with high reflecting sputtering aluminum, the sterilization efficiency were above 99.999%～99.999 9% when the atmosphere passed through the germicidal tubes. As a technical breakthrough, the Virobuster steritubes could overcome the problems of traditional air sterilization and decontamination technology, the air sterilization could reach LOG-6 sterilizing effect. By analyzing of the workings principles and structure characteristics of Virobuster steritube, the optimization design of air sterilization and decontamination is set up for our public buildings air and hospital sterile room.

Swine influenza A virus belongs to the family of Orthomyxoviridae,which can cause upper respiratory tract infection in swine,avian and human.The influenza A virus that could not be sub-typed was detected from the respiratory tract specimens of patients with influenza-like symptoms in March 2009 in USA and Mexico,followed by identification of a novel swine-origin influenza A (H1N1) virus (S-OIV),which led to an epidemic outbreak in the above two countries and then spread all over the world,causing quite a few deaths.S-OIV differs from other influenza viruses in genotyping,antigenic characteristics,propagating characteristics,pathogenic characteristics and therapies.

Objective To establish a GC-MS method for the analysis of the chemical components of essential oil from Fructus Ponciri Trifoliatae Immaturi.Methods The essential oil was extracted by steam distilyation ,then separated by capillary gas chromatography. The components from essential oil were identified by comparing with related standard chromatogram and their amount were determined by normalization method.Results Twenty-one components have been identified from Folium Fructus Ponciri Trifoliatae Immaturi and their amount accounted 99.04 %of total essential oil. The main components are limonene (55.68 %),?-myrcene(21.96 %),?-Pinene(5.17 %),cis-caryophyllene(4.61 %)and ?-Ocimene (1.71 %).Conclusion This method is reliable and effective and can be applied to analyze the essential oil components extracted from Fructus Ponciri Trifoliatae Immaturi.