ObjectiveTo investigate the levels of serum interleukin -17(1L-17) and tumor necrosis factor -alpha( TNF- α ) and synovium pathological changes in patients with rheumatoid arthritis ( RA ) and its clinical significance.MethodsThe levels of serum IL-17 and TNF- α in 50 patients with active phase of RA ( active phase of RA group),50 patients with stable phase of RA ( stable phase of RA group ) and 50 normal controls (NC group) were measured by enzyme-linked immunospecific assay(ELISA),and the synovium was detected by pathological examination.ResultsThe levels of serum IL- 17 and TNF- α in active phase of RA group [(42.60 ± 11.30),( 113.20 ± 13.11 ) mg/L]were significantly higher than those in stable phase of RA group [( 19.60 ± 5.75),( 14.50 ± 5.33) mg/L]and NC group [(7.40 ± 3.32),( 10.90 ± 2.24 ) mg/L](P ＜0.01 ),the level of serum IL-17 in stable phase of RA group was significantly higher than that in NC group ( P ＜ 0.05),but the level of TNF- α had no significant difference between stable phase of RA group and NC group ( P ＞ 0.05).The synovium was significantly different between RA patients and NC group.Conclusion IL-17 and TNF-α are more sensitive index for detecting the activity of RA and may play important rolesinmonitoring the therapeutic effects and prognosis of RA.

Objective To estabolish the ALDH2 Glu504Lys genotyping technique for oral swab sample type based on PCR-gold magnetic nanoparticle chromatographyl.Methods According to the ALDH2 Glu504Lys gene the specific primers were designed by ARMS-PCR technique.Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR.60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography.The results were verified by sequencing.Results The detected 60 cases of oral swab samples contain 16 cases heterozygous mutation,wild type 42 cases,homozygous mutation 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results.Conclusion The Goldmag nanoparticles chromatographic detection technique of ALDH2 Glu504Lys buccal swabs system was established.With accuracy,fastness,reliability,it is worthy of clinical promotion.

Objective:To research the TLC identification method for Longdan Xiegan pills( honey pills) . Methods:TLC was used in the identification. The samples were extracted by 70% methanol with a heating reflux method, and then extracted by the agents with dif-ferent polarity, including petroleum ether, ethyl acetate and butanol. The petroleum ether part was detected by fluorescence at 365nm for Angelica sinensis, and 1% vanillin-sulfuric acid color reaction was used to detect Alisma orientale. The ethyl acetate part was determined by fluorescence at 365nm for Scutellaria baicalensis, and the butanol part was detected with chloroform-methanol-water (30∶12∶3) as the developing solvent for Bupleurum and Glycyrrhiza, and with acetone-ethyl acetate-water (6∶6∶1) as the developing solvent for gentiopi-croside, geniposide and liquiritin. Results:The developed TLC spots were clear with good separation, high specificity and promising re-producibility. Conclusion:The method can be exactly used in the qualitative identification and quality control of Longdan Xiegan pills ( honey pills) .

Many Chinese medicinal materials are difficult to be distinguished in characters resulting in easy confusion. In the pa-per, the basic concepts of three groups of easily confused Chinese medicinal materials ( schisandrae chinensis fructus and schisandrae sphenantherae fructus, phellodendri cortex and phellodendri amurensis cortex, akebiae caulis and clematidis armandii caulis) were illus-trated and the development of verification methods were reviewed in the respect of modern instrumental analysis technology. With the improvement of analysis technology, the identification of Chinese medicinal materials relies on experimental data instead of artificial ex-perience, which makes it more objective and accurate.

Objective: To establish a method for the content determination of naringin and neohesperidin in Weitong pills.Methods: An HPLC method was adopted.The determination was performed on an Agilent TC-C18 column with the mobile phase consisting of acetonitrile-water(20∶80)at the flow rate of 1.0 ml·min-1.The detection wavelength was set at 283nm.The column temperature was 30 ℃.Results: Naringin and neohesperidin respectively within the concentration range of 0.027-4.552 μg (r=0.999 9) and 0.029-4.016 μg(r=0.999 8) had good linear relationship with the peak area, the average recovery was 102.19% and 103.60%,and the RSD was 2.88% and 1.79%(n=9), respectively. Conclusion: The method is simple,accurate and reproducible, and suitable for the content determination of naringin and neohesperidin in Weitong pills.

Objective To investigate the effect of cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal therapy on biochemical indicators of postoperative intracranial infection, in order to improve the clinical diagnosis and treatment. Methods 70 cases with intracranial infection collected in Third Hospital of Beijing Armed Police Corps from February 2010 to April 2013 were as subject, and randomly divided into two groups. Control group(n=35) were given cerebrospinal lfuid replacement and ceftriaxone intravenously, observation group(n=35) were given cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal injection. The clinical effects and biochemical indicators were observed after treatment in two groups. Results In control group, the cure rate was 22.86%and total efifciency was 77.14%. In observation group, the cure rate was 37.14% and total efficiency was 91.43%. The differences between two groups were statistically significant (P<0.05). The differences of leukocytes, glucose, protein, intracranial pressure in two groups after treatment were also statistically signiifcant(P<0.05). Conclusion Cerebrospinal lfuid replacement combined with vancomycin and dexamethasone intrathecal injection therapy can increase intracranial infection.

Objective To explore the inhibitory effects of lobaplatin and cisplatin and their regulation of apoptosis-related genes in ovarian cancer cells in nude mice.Methods SKOV3 cells were implanted into nude mice.In monotherapy treatment study,the nude mice bearing human SKOV3 cells were randomly divided into control,lobaplatin,and cisplatin groups,with 7 mice in each group.The mice in each group were received corresponding treatment.The volume of tumor and the weight of nude mice were measured three times per week,respectively.Tumor inhibitory rate was calculated.The protein expressions of bax and bcl-2 were detected by flow cytometry.Results The growth inhibitory rate was 47.2％ in lobaplatin group and 42.8％ in cisplatin group,without significant difference between two groups (P ＞ 0.05).The expression of bcl-2 was decreased but the bax was increased in lobaplatin and ciaplatin groups compared to the control group.Conclusions Lobaplatin can significantly inhibit the growth of ovarian cancer cells,induce apoptosis by down-regulation of bcl-2 and up-regulation of bax.

Objective To study the role of human MutS homolog 2 (hMSH2), a newly identified protein ligand that was recognized by Vγ9δ2 T cells , in innate anti-gastric carcinoma immunity .Methods Flow cytometry and laser confocal microscopy were used to identify hMSH 2 that ectopically expressed on gas-tric carcinoma cell line 803.An anti-hMSH2 antibody was used to block hMSH2 to evaluate its effects on the cytotoxicity of Vγ9δ2 T cells and their cytokines secretion .Subcellular distribution of hMSH 2 in gastric car-cinoma tissues was examined by tissue microarray immunohistochemistry analysis .Results Ectopic mem-brane expression of hMSH 2 was observed on 803 cells at a relatively high level .Vγ9δ2 T cells blocked with specific anti-hMSH2 antibody showed a decreased cytotoxicity and a reduced IFN-γbut an increased TNF-αsecretion.Ectopic expression of hMSH2 was found in various types of gastric carcinoma tissues at different stages.Enhanced expression of hMSH2 was detected in specimens collected from patients with chronic super-ficial gastritis.Conclusion Ectopically expressed hMSH2 served as a stress-induced endogenous ligand which could promote the cytotoxicity of Vγ9δ2 T cells against gastric carcinoma cells and enhance their IFN-γsecretion.hMSH2 played an essential role in innate anti-gastric carcinoma immunity .

Objective To construct human lung cancer cell model with human MutS homologous protein 2 (hMSH2) overexpression for exploring the effect of hMSH2 molecule in the cytotoxicity of γδ T cell against lung cancer cells.Methods hMSH2 coding sequence was cloned by PCR for construction of recombinant vector which over expressed hMSH2-EGFP fusion protein using homologous recombination.The recombinant vector was transfected to lung cancer cell line NCI-H520 to construct human lung cancer cell model overexpressing hMSH2 molecule.The expression of hMSH2 molecule in NCI-H520 was detected by Western blot.The expression of hMSH2 on the cell membrane was measured by flow cytometry.Cytotoxicity of expanded γδ T cells from peripheral blood mononuclear cells against NCI-H520 cells was detected by LDH release assay in vitro.Results hMSH2 coding sequence (2805 bp) was cloned and the result of restriction endonuclease digestion of Fugw-hMSH2 recombinant vector was accordance with the anticipated objective strip size.Exogenous hMSH2-EGFP fusion protein was expressed in NCIH520 cells.The level of hMSH2 molecule on the surface of NCI-H520 cells with overexpression of hMSH2 was significantly increased (P＜0.001).Cytotoxicity of γδ T cells against NCI-H520 cells with overexpression of hMSH2 was significantly increased compared to the wild type NCI-H520 cells (P＜0.05).Conclusions Lung cancer cell model that overexpresses hMSH2 molecule is successfully constructed,hMSH2 molecule on the cell membrane is increased and the cytotoxicity of γδ T cells against lung cancer cells is enhanced.

To obtain non-pathogenic rabies virus glycoprotein(RV-G),we expressed RV-G in Saccaromyces Cerevisiae(S.cerevisiae).In our study,tat-G fusion gene was cloned into the expression vector pYes2.0,which allows expression of a foreign gene in the yeast cells under the control of GAl1 promoter.Transforma-tion was performed by using lithium-treated yeast cells and several Ura+-tranformants were isolated.Ac-cording to the relative mobility in SDS-PAGE,we know probably two forms(designated as yGI and yGⅡ) of RV-G analogues produced in S.cerevisiae,their molecular weights were estimated as 66 kD and 56 kD,respectively.On the other hand,there was a specific band about 56 kD shown in western blot result.Com-bining precursors’ achievements,we will draw a conclusion that trans-membrane domain(TD) and cyto-plasmic domain have a negative regulation on RV-G antigen immunogenicity in S.cerevisiae.