Objective To explore the T‐SPOT .TB technology in latent tuberculosis infection (LTBI) who immunosuppres‐sive therapy results in screening for latent tuberculosis infection prevention and control to provide a new basis .Methods Applica‐tion of T‐SPOT .TB kit 162 immunosuppressed patients need to be applied to detect M .tuberculosis‐specific T cells ;while doing all cases tuberculin (TST ) skin test ;of which 28 cases of T‐SPOT .TB‐positive patients before screening technique using anti‐TNF‐αbiologics were given prophylactic treatment of anti‐TB drugs for 4 months and followed a year .Results The positive rates and ac‐curacy rate of T‐SPOT .TB assay were 36 .4% and 94 .9% ,while the positive rates and accuracy rate of TSTs were 28 .4% and 69 .6% .The difference between T‐SPOT .TB assay and TST were statistical significance(P < 0 .05) .Through our 28 cases of T‐SPOT .TB positive screening technology ,prophylactic anti‐TB drugs to treat patients for 4 months and 1 year of follow‐up ,no case of tuberculosis occurred .Conclusion These results demonstrate that the performance of T‐SPOT .TB is better than the classic TST for detection of LTBI in patients receiving immunosuppressive therapy for treatment of systemic autoimmune disorders .The T‐SPOT .TB assay will be a useful tool in early and rapid diagnosis of latent tuberculosis infection .T‐SPOT .TB for LTBI patients di‐agnosed with prophylactic anti‐TB drug treatment is necessary ,has important clinical significance .

Beginning with the authors' experience in analytic-chemistry bilingual teaching,this article probes into some problems of analytic-chemistry in undergraduate education.And the authors give some beneficial suggestions in the teaching material construction,teachers training and teaching quality control.

The present situation and problems in experiment teaching of analytical chemistry were explained,and innovation was explored and practiced in the faculty of laboratory medicine,including the opitimization of teaching contents,teaching methods and score system evaluation reformation.

The academic teaching quality and experimental teaching quality of sanitary chemistry will be improved,through enriching the teacher with specialized knowledge and cultivating the creative ability of thacher and updating the teaching methods.

By evaluation of academic designed experiment performance in Bio-Analytic Chemistry from ten different evaluating indicators including literature search, experimental design, and experimental report and so on. The quantitative analysis results indicate the average is 87.8, the standard deviation is 5.0,the difficulty index is 0.88,the dipartite degree is 0.12 and the confidence coefficient of the assess is 0.9423. The designed experimental assessment can inspire the students’experimental goaheadism, cultivate their ability to analyze and solve problems,and promote their innovative consciousness and practice skills.

On the development of Bio-analytic chemistry curricula,to enhance the build- ing of teaching staff,improve the teaching content according to the developmental trend of model chemistry,adopt multiform methods to explore new teaching pattern,improve the teaching quality based on the manage,the course on Bio-analytic chemistry has became more systematized and standardized.

ABSTRACT:Objective To construct V-set and immunoglobulin domain containing 4 (Vsig4)nanobodies (Nbs) as specific macrophage probes so as to use them as molecular probes of macrophagocytes.Methods A nanobody phage library was generated by using peripheral blood lymphocytes isolated from an alpaca immunized with recombinant Vsig4 protein.After three rounds of selection against recombinant Vsig4.The Nbs were subjected to sequencing and genome alignment to obtain VHH sequence.Nbs were isolated and tested for Vsig4 specificity in an ELISA using recombinant Vsig4.The affinity capacity of Nbs was verified by the cell line stably expressing Vsig4. Results A nanobody phage library with an estimated 7.27 × 107 clones with 70% insertion was successfully constructed.Totally 1 3 6 Vsig4-positive clones were sequenced and aligned according to different CDR3 sequences. In summary,1 5 Vsig4 nanobodies were obtained and grouped into 3 different CDR3 epitopes.The affinity of representing nanobody and Vsig4 was analyzed via ELISA;Nb1 1 9 showed the highest affinity against both recombinant and native Vsig4.Conclusion We successfully constructed and screened Vsig4 specific nanobody number 1 1 9 with high affinity and specificity.It can help with macrophage detection and in vivo monitoring.

Objective:To determine the in vitro release of two kinds of Changankang Pellets (Radix Astragall, Rhizama coptid) coated with Surelease and Eudragit respectively and verify their colon-targeted release in vivo through X-ray photography. Methods: Changankang Pellets were made by mixing the medical BaSO 4 powder and appropriate drugs together according to original produce and coat methods. After determining their in vitro release, they were tested in vivo by 2 volunteers. X-ray films were taken after appropriate time and analyzed to determine the location and status of these pellets inside healthy objects. Results: Pellets made by this way can carry drugs through stomach, intestine and targeted-release them in colon; the coating combined with pH-dependent and time-dependent Eudragit thin layer has better results. Conclusion: X-ray radiography applying BaSO 4 powder could be taken as a quality control method for colon-targeted release drugs. The preliminary results showed that Changankang Pellets met the requirements of colon-targeted release.

Teachers introduced the means of the scientific thinking and scientific research prac-tice in the curriculum of proteomics in order to train the postgraduates to joint the theory and the practical research work in the curriculum course. For example, to cultivate students to truly understand the 2-DE protein separation technique, teachers used the various proteomic profile obtained from our research to points out its problems and limitations based on the classical 2-DE, then compared the improved 2-DE protein separation chromatography with the classical 2-DE graphic and analyzed what is the problem and how to solve them in the practical work. To improve the ability of scientific thinking for postgraduates , teachers took the tumor as the teaching cases and asked the postgraduates to search the references of can-cer associated with their professional , to put forward the scientific problems based on carefully reading these references, to design their research project and to seek the answer to their issue. These new teaching methods were found to be beneficial in training scientific talents of medical postgraduates.

Background Tumstatin is the most active endogenous angiogenesis inhibitor,which has a marked inhibitory effect on pathological neovascularization,and Tum5 is an angiogenesis inhibitors fragment of fulllength tumstatin.Objective This study was to investigate the effects of adenovirus-mediated overexpression of recombinant Tum5 gene on the proliferation,migration and tube formation of human umbilical vein endothelial cells (HUVECs) in physiological status.Methods The empty adenoviral vector expressing green fluorescent protein (rAd-GFP) and the viral vector expressing recombinant Tum5 gene were constructed.The HUVECs cultured in RPMI1640 medium were divided into normal control group,empty vector group (rAd-GFP group) and Tum5 gene infection group (rAd-GFP-Tum5 group).The rAd-GFP and rAd-GFP-Tum5 adenoviral particles at the density of 1 × 1010/ml were added into the medium to infect the cells for 48 hours.The proliferation of the cells was assayed at 24,48 and 72 hours by cell counting kit-8 (CCK-8) to evaluate the proliferative rate;the migration number of the cells was detected at 48 hours after infection by Transwell chamber;the tube formation number of the cells were detected by Matrigel method.The concentration of vascular endothelial growth factor (VEGF) in cell supernatants was assayed by ELISA at 24,48,and 72 hours following adenoviral infection.Results The cultured cells showed green fluorescence in the rAd-GFP group and rAd-GFP-Tum5 group under the inverted fluorescence microscope,and the infection efficiency of rAd-GFP and rAd-GFP-Tum5 was 55.13％ and 50.31％,respectively.No significant difference was found in cell proliferative rate among normal control group,rAd-GFP group and rAd-GFP-Tum5 group both at 24 and 48 hours after infection (both at P＞0.05),and the cell proliferative rate was significantly lower in the rAd-GFP-Tum5 group than that in the normal control group and rAd-GFP group at 72 hours after infection (both at P＜0.01).The migration number of the cells at 48 hours after infection was 2 260.25-±930.44,2 370.00±441.06 and 723.75± 363.80 in the normal control group,rAd-GFP group and rAd-GFP-Tum5 group,showing a significant difference among the groups (F =8.524,P =0.008),and the migrated cells were evidently decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group and the normal control group (both at P＜ 0.01).The tube number at 48 hours after infection was 95.67±5.86,88.00±4.58 and 20.67±3.51 in the normal control group,rAd-GFP group and rAdGFP-Tum5 group,showing a significant difference among the groups (F=226.498,P＜0.01),and the tube number in the rAd-GFP-Tum5 group was significantly reduced in comparison with the normal control group and rAd-GFP group (both at P＜ 0.01).The considerably differences in VEGF concentration in the cell supernatants were found in different groups and various time points (Fgroup =73.260,P＜0.01;Ftime =73.477,P＜0.01),and VEGF concentration in the cell supernatants was significantly decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group at both 48 hours and 72 hours (both at P＜0.01).Conclusions The overexpression of the recombinant Tum5 can inhibit the proliferation,migration and tube formation of the HUVECs in physiological status,which may be associated with Tum-5-mediated down-regulation of VEGF protein in the cell supernatant.