Objective In this study,we measured the levels of urinary monocyte chemoattractant (MCP)-1 and interferon-γ-inducible protein (IP-10) and further analyzed their associations with clinical and pathological data in lupus nephritis patients in order to find the non-invasive biomarkers which canpredict disease activity.Methods MCP-1,IP-10,VEGF levels were measured in urine samples from 64 lupus nephritis patients and 20 healthy volunteers.Clinical disease activity was determined by SLEDAI and BILAG scores.The lupus nephritis patients were divided into two groups:active disease group (SLEDAI scores ≥ 10points,n=36) and non-active group (SLEDAI score＜10 points,n=28).Of all patients enrolled,37 patients had a concomitant kidney biopsy performed at the time of urine collection.The predictive performance of uri-nary MCP-1 and IP-10 for renal flare,the Student's t test,Mann-Whitney U test,Chi-square test,and re-ceiver operating characteristic (ROC) curves were constructed for analysis.Results The urinary MCP-1 and urinary IP-10 levels of the active group was significantly higher than that of the non-active group [MCP-1672.39(318.05,2 554.23)pg/ml vs 152.52,(55.61,330.44)pg/ml,Z=-4.717,P＜0.01; IP-10 (38±19) pg/ml vs (22±16) pg/ml,t=3.576 P＜0.01].The level of urinary MCP-1 was positively correlated with the levels of hematuria and 24 hours protein quan-tification,as well as the scores of SLEDAI and BILAG (rbemahuria=0.570,P=0.000; r24hpro=0.569,P=0.000; rSLEDAI=0.600,P=0.000; rBILAG=0.606,P=0.000),and it was also positively correlated with the scores of cellular crescent,wire loop,and AI (rCC=0.405,P=0.015; rwire loop=0.430,P=0.014; rAI=0.352,P=0.003),while nega-tively correlated with the level of C3 and plasma albumin (rc3=-0.564,P=0.000; ralb=-0.587,P=0.000).It had no correlation with the scores of wire loop and CI (P＞ 0.05).The level of uIP-10 was positively correlated with the protein quantification in 24 hours and the scores of SLEDAI and BILAG (r24hpro=0.305,P=0.018; rSLEDAI=0.334,P=0.009; rSILAG=0.496,P=0.000),while negatively correlated with the level of C4 (rC4=-0.301,P=0.016).The R0C curve of uMCP-1 to predict the activity of SLE showed that its specificity was 75.0％,sensitivity was 83.3％,and the area under the ROC curve was 0.85±0.05.The ROC curve of urinary IP-10 to predict the activity of SLE showed that its specificity was 50.0％,sensitivity was 97.2％,its area under the ROC curve was 0.74±0.06.The ROC curve of urinary MCP-1 to predict renal flare shows that its specificity was 45.5％,its sensitivity was 100％,and the area under the ROC curve was 0.74±0.80.The ROC curve of urinary IP-10 to predict renal flare showed that its specificitywas 36.4％,its sensitivity was 73.3％,and its area under the ROC curve was 0.49 ±0.10.Conclusion Urinary MCP-1 and urinary IP-10 predict renal flare in patients with lupus nephritis.Furthermore,urinary MCP-1 is a more specific and sensitive forecaster of renal flare in patients with a history of lupus nephritis than urinary IP-10.

Objective To investigate whether expression quantity of HLA-B27 and its subtypes associated with incidence of AS,the gene expression of HLA-B27 and its subtypes were detected in patients with AS.Methods 120 cases patients sus-pected with AS and 50 healthy subjects were enrolled in the study.Main demographic and clinical characteristics of the sub-jects were collected.Meanwhile total RNA was isolated from peripheral blood and real time RT-PCR was used to measure the quantitative expression of HLA-B27 gene.Besides RT-PCR,sequence-based typing (SBT)method was used to confirm the HLA-B27 subtype.All experimental data were analyzed by SPSS17.0 software and P values <0.05 were considered to be significant.Results Firstly,there were 55 subjects were finally diagnosed as AS patients among the 120 patients suspec-ted with AS.There were 53 subjects whose HLA-B27 was positive (96.36%)in 55 patients with AS.It showed that there was a correlation between BASDAI and expression quantity of HLA-B27 gene (r=0.845,P =0.000).Five subtypes were found and which were HLA-B27∶04 subtype (29/53,54.72%),HLA-B27:05 subtype (20/53,37.74%),HLA-B27 ∶02 subtype (2/53,3.77%),HLA-B27∶03 subtype (1/53,1.89%)and HLA-B27∶07 subtype (1/53,1.89%),respectively. Among the 50 healthy subjects,there were only one kind of subtype (2/50,4%),which was HLA-B27∶04.There were no statistical difference in the age (t=0.711,P =0.480),sex (χ2 =0.880,P =0.348),family history (χ2 =0.011,P =0.916) and treatment (χ2 =0.113,P =0.736)between the HLA-B27∶04 and HLA-B27∶05 subtypes.Conclusion HLA-B27∶04 and HLA-B27∶05 were primary subtypes in AS patients which HLA-B27 positive.There was a correlation between gene expression quantity of HLA-B27 and AS disease activity index.

Objective:To investigate the in vitro effects of platelet-rich plasma(PRP) on human periodontal ligament fibroblasts(PDLFs). Methods: Various concentrations of PRP (10, 50, 100, 200, 300, 500 ml/L) were applied to primary cultures of human PDLFs. MTT assays were utilized to assess cell proliferation ability. Migration was determined by assessing the cell response to a concentration gradient with Transwell chamber. Differentiation was assessed using alkaline phosphatase (ALP) kit. Results: A beneficial effect on proliferation was observed, especially in response to 200 ml/L PRP.PRP had stimulatory effects on the migration of human PDLFs. PRP facilitated differentiation of PDLFs. Conclusion: PRP can exert a positive effect on human PDLFs,but this effect is concentration specific, while higher concentrations is not necessary to result in optimal outcomes.

ObjectiveTo find specific biomarkers related to HAART treatment in plasma samples of AIDS patients for clinical therapeautic efficacy evaluation and guidance for the prognosis of HIV treatment.MethodPlasma samples of AIDS patients were collected from Infectious Disease Department 1 of Shanghai Public Health Clinical Center in June of 2008 to February of 2009,including 11 successfully HAART treated cases (HIV load ＞ 50 copies/ml) and 11 unsuccessfully HAART treated cases (HIV load ＜50 copies/ml).Patients' age ranged from 22 to 63.Plasma samples were treated by Bio-rad AurumTM Serum Protein Mini Kit to remove high abundant proteins:albumin and immunoglobulin were removed.The treatedplasmaproteinswereseparatedbytwo-dimensionalelectrophoresisandanalyzedby electrophoretogram using Imagemaster software to find differentially-expressed proteins related to therapeutic efficacy.After digestion by trypsin,the differentially-expressed proteins were identified by online reversed-phasenano-flow liquid chromatography coupled with electrospray ionization ion trap mass spectrometry.ResultsLow abundant proteins were efficiently enriched after the AurumTM Serum Protein Mini Kit treatment.Six differentially-expressed proteins were detected while comparing successfully and unsuccessfully HAART treated group.These proteins were accurately identified by tandem Mass spectrometry (MS), including serum transferrin, serum β-fibrinogen, etc.ConclusionsOur proteomic research revealed that the differentially-expressed proteins such as transferrin,which is related to plasma virus loading in AIDS patients in the process of treatment,might be potential biomarkers evaluating HAART therapeutic efficacy.

A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of lopinavir and ritonavir in human plasma. Analytes were separated from plasma by a combination of alkalinized protein precipitation and liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a Agilent ZORBAX Eclipse XDB-C18 column with the mobile phase consisted of methanol-0.1% formic acid in water (80:20). A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the positive ion mode. Quantification was performed using multiple reaction monitoring (MRM) of the transitions m/z 629.6 --> 155.2, m/z 721.4 --> 268.2, and m/z 515.2 --> 276.2 for lopinavir, ritonavir and telmisartan (internal standard), respectively. The method showed a good linearity in a concentration range of 62.5 - 10000 ng mL(-1) for lopinavir, and 12.5 - 2000 ng mL(-1) for ritonavir. The lower limits of quantification were 15 pg mL(-1) and 8 pg mL(-1) for lopinavir and ritonavir, respectively. The intra- and inter-day precision was less than 15% and the absolute recovery was above 75%. This method was selective and rapid, sensitive for investigating blood drug concentrations in clinics.

Objective To analyse clinical application of hepatitis B virus core antibody(HBcAb)detected by using the chemilu-minescence microparticle immunoassay.Methods A total of 1 6 830 specimen with positive HBcAb detected by using the two pairs of semi-hepatitis test from January 2012 to November 2014 were collected,and divided into three groups according to the cut off in-dex(COI)of detection results of HBcAb,including group 1.0-<9.0,group 9.0-<1 1.0 and group COI≥1 1.0,and detection re-sults were statistically analysed.The hepatitis B virus(HBV)DNA test was carried out in specimen with negative hepatitis B surface antigen(HBsAg)and hepatitis B surface antibody (HBsAb)and COI≥1 1.0.Results The detection rate of HBsAg(+)HBsAb(-) (13.84%)was significantly higher than other expression patterns in group ≥1 1.0(P <0.05).There was no statistically significant differences in positive rate among all expression patterns of HBsAg and HBsAb in the group 9.0-<1 1.0(P >0.05).The detec-tion rate of HBsAg(+)HBsAb(-)of group 9.0-<1 1.0 was significantly lower than that of the other two groups(P <0.05).A total of 304 specimen were HBsAg(-)HBsAb(-)and COI≥1 1,among them 64 specimen were HBV DNA postive and the posi-tive rate was 21.0%.Conclusion In the detection of HBcAb,COI≥1 1 and 1.0-<9.0 could be reference indicators for diagnosiing current and past HBV infection respectively,which should be combined with other laboratory indicators of HBV clinical data for comprehensive analysis.

Objective:To explore the effect of ionizing radiation on apoptosis of lung cancer H460 cells after ATRX was silenced by RNAi and its mechanism.Methods:The lentivirus expression vectors targeting ATRX were transfected into the 293T cells,and the lung cancer H460 cells were infected with lentivirus twice,and the ATRX silenced cell model was obtained after puromycin positive screening,then they were named as sh-ATRX1-H460,sh-ATRX2-H460,and sh-ATRX3-H460 cells;the sh-control-H460 cells were regarded as control cells.The cells were divided into sh-control-H460 group and sh-ATRX3-H460 group,accroding to the silencing results and were irradiated by 0,2 and 8 Gy X-rays.The expression levels of ATRX,poly(ADP-ribose) polymerase 1(PARP1),and caspase-3 proteins were measured by Western blotting method;the apoptotic rate was measured by flow cytometry and AnnexinⅤ-FITC/PI kits.Results:The lung cancer cell model of sh-ATRX3-H460 silenced by ATRX was obtained successfully.After 2 and 8 Gy X-ray irradiation,compared with before irradiation,the expression level of ATRX protein in sh-control-H460 group was increased,while there was no expression of ATRX protein in sh-control-H460 group;compared with before irradiation,the apoptotic rates of cells in two groups were increased(P<0.05 or P<0.01);the apoptotic rate in sh-ATRX3-H460 group was significantly higher than that in sh-control-H460 group after 8 Gy X-ray irradiation (P<0.01).The expressions of cleaved PARP1 in the cells in both two groups after 2 Gy and 8 Gy X-ray irradiation were increased and showed similar rule.The expression level of procaspase-3 protein in sh-control-H460 group had little change,and it was increased significantly in sh-ATRX3-H460 group after 8 Gy X-ray irradiation.Conclusion:ATRX silencing can be achived by RNAi,then the silencing could increase the apoptosis induced by irradiation and its mechanism may be related to the PARP1-caspase-3 pathway.

Research of co-authorship network can reveal the scientific research collaboration network and can thus help us to have an understanding of it. Graphic database and Neo4J were described in detail due to the limitations of relationship database in processing the data of co-authorship network. Graphic database Neo4J-based research and practice of co-authorship network were analyzed with the Institutional Knowledge System of AMMS that we were involved in its construction as an example, and the advantages of Neo4J-based co-authorship network were summarized.

This study aimed at analyzing the medication regularity based on differentiation in traditional Chinese medicine (TCM) for losing weight using text mining technique.All the references over losing weight were retrieved in CNKI,Wangfang Database,VIP Database and Pubmed.The drugs from the references were classified in accordance with drug property,drug flavor,channel tropism and drug efficacy.Frequency and constituent ratio of a single drug in TCM prescriptions for losing weight were put into analysis using chi square test and factor analysis to find out the medication regularity.It was found that the properties of TCM drugs in the prescriptions contained both cold and warm,while the flavors of the drugs involved pungent,sweet and light.The channel tropism of the drugs mainly belonged to spleen meridian,liver meridian,stomach meridian and lung meridian.They were mostly tonic,relieving,blood-activating,qi regulating,inhibiting-damp and antipyretic drugs.Through factor analysis we found that the common formula compatibilities were concluded as:cassia seed,lotus leaf,hawthorn,salvia miltiorrhiza,polygonum cuspidatum and radix polygonum multiflorum;capillary artemisia,epimedium herb,stephania tetrandra and ligusticum wallichii;dried tangerine peel,pinellia ternata and poria cocos;plantain seed,pericarpium arecae and selfheal;paeonia lactiflora,angelica sinensis,scutellaria baicalensis and ligusticum wallichii;poria cocos,cassia twig,atractylodes and glycyrrhiza;immature bitter orange and bark of magnolia;radix bupleuri,lycium chinensis and jujube;Chinese yam and coix seed;and astragalus,pueraria lobata and polygonatum.In conclusion,formula compatibility mainly combined syndrome differentiation with disease differentiation for the treatment of obesity in clinic,using the drugs belonging to liver meridian,spleen meridian,stomach meridian and lung meridian with the flavors of sweet,bitterness or pungent and the nature of both warm and cold.

OBJECTIVETake human oral squamous cell carcinoma Tca8113 as experimental model, and study the anti oral squamous cell carcinoma activity of high mobility group chromosomal protein N2 (HMGN2) molecule.

METHODSTrain a large number of recombinant human HMGN2 expression vector Escherichia coli BL21. HMGN2 was expressed under isopropyl-1-thio-beta-galactopyranoside (IPTG) induction and purified by B-PER GST Fusion Protein Purification Kit. A variety of concentrations HMGN2 were added to cell culture medium, cells were tested by MTT, Hoechst 33342 fluorescence staining, flow cytometry assay and Western-blot.

RESULTSMTT results proved that HMGN2 could significantly inhibit human oral squamous cell carcinoma Tca8113 growth. Hoechst 33342 fluorescence staining, flow cytometry assay test and Western-blot proved HMGN2 could make Tca8113 cells morphological change, make Tca8113 cells block in S period of cell cycle and strongly promote Tca8113 cells to apoptosis.

CONCLUSIONHMGN2 can promote apoptosis of oral squamous cell carcinoma cells.