Objective To investigate the effects and mechanisms of 5-aza-2'-deoxycytidine (5-Aza-CdR) on endometrial cancer cell.Methods In vitro experiments of 5-Aza-CdR were done using human endometrial cancer cell line HEC-1B.Evaluation of cellular proliferation and apoptosis was ascertained respectively using trypan blue exclusion and flow cytometry.RT-PCR and methylation specific PCR(MSP) was done to detect the expression of RASSF1 A mRNA and methylation status of RASSF1 A promoter of HEC-1B cell line.Results (1) The status of cellular growth and apeptosis of HEC-1 B cell line:the growth inhibition effects of 5-Aza-CdR on HEC-1B cell line were both concentration-dependent (P ＜ 0.01) and time-dependent(P ＜0.01),as well as the apoptosis rate of HBC-1-B cell line depended on the dose of 5-Aza-CdR obviously(P ＜0.01).(2)The expression of RASSF1A mRNA of HEC-1B cell line:RASSF1A mRNA was expressed in HEC-1B cell after 5-Aza-CdR treatment,but it was undetectable before the treatment.In the groups with different concentration of 5-Aza-CdR (0.05,0.1,1,5,10 nmol/ml),the expression of RASSF1A mRNA was respectively 0.074±0.004,0.105±0.004,0.167±0.006,0.334±0.005,0.484±0.007,which were remarkably different from the group without 5-Aza-CdR(the expression of RASSF1A mRNA was 0;P ＜ 0.01).(3) The hypermethylation of RASSF1A promoter of HEC-1B cell line:the hypermethylation of RASSF1A promoter was detected in HEC-1B cell line.The status of hypermethylation was decreased after treatment with 5-Aza-CdR of 0.05,0.1,1,5 nmol/ml,meanwhile,both methylation bands and demethylation bands were observed by methylation specific PCR.After the treatment with 5-Aza-CdR of 10 nmol/ml the hypermethylation was absent absolutely.Conclusions (1) In HEC-1B cell line,5-Aza-CdR can inhibit cell proliferation and induce cell apopotosis.(2) 5-Aza-CdR can renew the expression of RASSF1A mRNA of HEC-1B cell line and reverse the hypermethylation of RASSF1A promoter.

Objective To compare the influence of cryopreservation duration on quality of rozen-thawed embryo and clinical outcome in frozen-thawed transfer cycles. Methods A retrospective study was performed to evaluate the clinical outcomes and quality of rozen-thawed embryo of frozen embryo transfer(FET)cycles includ-ing freezing cycles and vitrification cycles. FET cycles were divided into 4 group acorrding to the storage duration of vitrified embryos:1~6 months group ,7~12 months group ,13~24 months group and ≥ 25 months group. Result There was no significant difference in recovery rate ,clinical pregnancy rate and abortion rate (98.1%, 99.0%,96.8%&95.7%;54.4%,55.4%,49.6%&51.5%;0.7%,8.9%,9.4%&11.1%;P>0.05). The embryos recovery rate and clinical pregnant rate after vitrification were significantly higher than programmed freezing (P<0.05). The blastocyst recovery rate in group A was significantly higher than the cleavage stage embryo recov ery rate in group B(P<0.05). Conclusions Vitrification has better effects on cryopreservation and clinical outcomes. Blastocyst may be the best stage for vitrifying and freezing of embryo.

Objective:To prepare urokinase targeted thrombus microbubbles combined with low frequency ultrasound to dissolve thrombus in rabbit femoral artery, and to explore the mechanism of thrombolysis through the change of urokinase concentration.Methods:Twenty-four rabbits with thrombosed femoral artery were randomly divided into four treatment groups: urokinase alone (UK) group, ultrasound with non-targeted microbubble and urokinase (US+ M+ UK) group, platelet-targeted microbubble with urokinase (R+ UK) group, ultrasound with platelet-targeted microbubble and urokinase (US+ R+ UK) group. Thrombus-targeted microbubbles were injected through the ear vein and irradiated by ultrasound for 30 minutes, and the pulsed Doppler blood flow meter was used to continuously monitor the blood flow at 0 min, 10 min, 20 min, 30 min, 60 min, 90 min and 120 min after injection. At the same time, the changes in urokinase concentration were monitored, and the characteristics of blood flow and urokinase concentration changes were analyzed.Results:UK and US+ M+ UK groups failed to show recanalization at 120 minutes after treatment.In contrast, the US+ R+ UK and R+ UK groups both achieved persistent recanalization( P<0.001). Compared with the basal state. Both the R+ UK group and R+ UK+ US group showed a decrease at 60 minutes, and the difference was statistically significant( P<0.05). Conclusions:In the thrombolysis process of low-frequency ultrasound combined with a targeted contrast agent carrying urokinase, the targeting can make the local urokinase concentration of thrombus reach the highest level. At the same time, ultrasound and targeted microbubbles promote urokinase to enter the thrombus, and finally achieve the strongest thrombolytic effect.

Objective To evaluate the clinical and radiographic characteristics and function of erosive hand osteoarthritis (EOA) patients. Methods Data were obtained from 19 patients with EOA, including their social conditions, clinical conditions, radiographic scores and hand function evaluation. The number of hand osteoarthritis (HOA) patients was 312. The control group consisted of non-EOA patients with hand osteoarthritis with a ratio of 4:1 to EOA patients. A non-parameter test analysis was performed. All data were analyzed by SPSS 23.0 statistical analysis, t test, χ2 test, Fisher exact probility and Spearman's correlations analysis were used for statistical analysis. Results Totally data of 19 patients were collected. Eighteen were female. Onset age was (56±8). Average duration was 56 (12~120) months. FIHOA scores of all the EOA patients were at least 5. All the erosions of 39 joints were characteristically central and erosive changes in 7 joints (18%) showed up as gull-wing. Among 39 erosive joints, including 12 (31%) E and 27 (69%) R, 34 (87%) distal interphalangeal joints were involved. Data analysis found out that EOA patients had longer disease duration (Z=2.610, P=0.009), more severe K-L level (44 ±11 vs 26 ±7, t=7.134, P<0.01), higher AUSCAN total score (28±6 vs 21±7, t=3.781, P<0.01) and higher AUSCAN function score (18±6 vs 12±6, t=4.042, P<0.01). The differences of ESR and CRP were not significant between EOA and non-EOA patients. Conclusion Erosions seen in EOA patients are centrally located gull-wing in the DIP joints. EOA patients have longer duration, more severe radiographic damage and worse joint function.

Objective To investigate the relationship between the expressions of VEGF,ENS, MVD and arteriosclerosis (AS) plaque in atherosclerosis obliterans (ASO) patients. Methods Immunohistochemical staining was used to determine the expression of VEGF, ENS and MVD value in the atherosclerosis plaque of ASO patients hospitalized from October 2007 to August 2010 in our hospital (n =30),and the femoral arterials of control were collected from 22 amputation after traumatic injury. Results The expressions of VEGF (2. 24 ±0.31 vs. 1.87 ±0.27,t =3.58),ENS (1.84 ±0.41 vs. 1. 56 ±0. 38, t =2.09) and MVD value (8.79 ±2.46 slip/HP vs. 6. 05 ±1.68 slip/HP, t =3.64) were significantly higher in the AS plaque of ASO patient than control (P <0.01 or P < 0. 05). We found positive correlations between the expression of VEGF, VEGF/ENS with MVD (r = 0. 391, P < 0. 01; r =0. 583, P < 0. 05) , while negative correlation between the expression of ENS with MVD (r=-0. 328,P <0. 05). Conclusion VEGF promotes the angiogenesis in atherosclerosis plaque,while ENS shows opposite effect The comprehensive performance of them is promoting angiogenesis.

Objective To investigate the expression levels of DNA methyltransferases 1, 3A and 3B in CD4+ T cells of MRL/lpr mice, and explore their relationship with the expression levels of methylation-sensitive genes (ITGAL, CD70). Methods CD4+ T ceils were isolated from spleens of 16-week-old MRL/lpr and BALB/c control mice by anti-CIM antibody labeled magnetic beads. Transcription levels of DNA methyhransferases 1, 3A and 3B and methylation-sensitive genes(ITGAL, CD70) were measured by real-time mice when compared with BALB/c control mice and the difference was significant (P<0.05), while the expression of DNMTI and DNMT3A showed a tendency of decrease (P>0.05). The mRNA expression of CD70 was significantly higher (P<0.01), but the expression of ITGAL had no significant difference between the two P<0.01 ). Conclusion Decreased expression of DNMT3B may attribute to the elevated expression of methylation-seusitive gene CD70, thus lead to the dysfunction of CD4+ T cell and play an important role in the pathogenesis of SLE.

Objective To explore the association between the signal transducer and activator of transcription 4 (STAT4) gene polymorphism and Chinese Han systemic lupus erythematosus patients. Methods Pyrosequencing technique was used to genotype the 3 single nucleotide polymorphisms (SNP) in the samples of patients and normal individuals. Results Not only these 3 SNPs, but also the haplotypes composed by them showed significant difference between the SLE patients and normal individuals[rs11889341:P=0.012 02, OR (95%CI)=1.22( 1.044～1.424); rs7574865:P=0.003 454, 0R(95%CI)=1.25(1.076～1.451 ); rs8179673: P=0.004 275, OR (95%CI)=1.274 (1.079～1.505)]. Conclusion rs11889341, rs7574865 and rs8179673 of STAT4 are associated with the pathogenesis of Chinese Han SLE, and the STAT4 gene is a susceptibile gene for SLE in a few racial cohorts.

Objective To discuss the clinical features of behavioral and psychological symptoms of demen-tia(BPSD)and the relation between different sub-clinical syndromes and cognition. Methods One hundred and sixteen dementia patients were assessed with neuropsychiatric inventory and mini-mental state examination (MMSE)and made factor analysis according to DSM-IV-R. Results Twelve common behavioral and psychological symptoms could be further divided into five sub-syndromes,including disinhibition behavior,psychosis,agitation, emotion and apathy factors. MMSE total score and years of education entered regression equation of apathy factor (P<0.05). Conclusion BPSD can be divided into five factors and apathy factor are related with cognitive function.

Objective:To clone and express Mycobacterium tuberculosis(Mtb) Rv2460c gene(encoding ClpP2 protein),and evaluate the immunogenicity of its coding product. Methods: The recombinant plasmid of pET32a (+) vector-ClpP2 that H37Rv Rv2460c gene was cloned into the plasmid pET32a(+)vector,was transformed into E. coli BL21(DE3)and induced expression by IPTG,then purified by affinity chromatography. The recombinant protein was confirmed by SDS-PAGE and Western blot. The analysis of immunogenicity of Mtb ClpP2 and its epitope prediction were performed by bioinformatic methods. The antibody levels of polyclonal antibody titer against ClpP2 protein in rabbits and TB patients′ serum were detected by ELISA. Results: The recombinant ClpP2 protease was expressed as inclusion bodies in E. coli. The purity of purified protein was 93% by bandscan software analysis. The bioin-formatics analysis shows Mtb ClpP2 protein has multiple preponderant B cell and T cell epitopes. Rabbit antiserum titer was 1∶64 000;Serum anti-ClpP2 antibody levels in TB patients was higher than that in healthy control subjects. Conclusion:The recombinant ClpP2 protein was purified, and specific Rabbit anti-ClpP2 polyclonal antibody was prepared successfully. Experiment and bioinformatic information studies showed that Mtb ClpP2 protease has strong immunogenicity.

Objective To study the clinical value of orbital magnetic resonance imaging(MRI) in diagnosis and making a judgment about the curative effect of glucocorticoid in patients with Graves' ophthalmopathy (GO).Methods A total of 106 patients with GO were divided into active GO group or inactive GO group according to whether clinical activity score(CAS) ≥3 or not,respectively.T2 relaxation times(T2RTs) (ms) and areas(mm2) of four extra-ocular muscles (EOMs) (superior,inferior,medial,and lateral rectus) were directly measured by MRI T2-Mapping.Correlation analysis of T2 RTs and areas with eyeball motility,diplopia,exophthalmus,visual acuity and intraocular pressure (IOP) were also performed.The T2RTs and areas of EOMs before and after intravenous methylprednisolone(iv MP) pulse therapy were recorded,respectively.Results The mean T2RTs (88.9 ± 13.8)and mean areas(58.1 ± 23.8) of EOMs in active GO were significantly greater than those in inactive GO (80.6 ± 10.0and 46.2 ± 18.6,respectively),bing both significantly greater than those in volunteers with normal eyes(76.2 ±4.7and 30.3 ± 6.1,respectively) (P＜0.01).Maximal T2RTs and maximal areas of EOMs showed significant positive correlation(P＜0.01),and both were positively correlated weakly with CAS,reduced movement of eyeball,diplopia,exophthalmus and loss of visual acuity (P＜0.01).On the contrary,maximal T2RTs and maximal areas of EOMs showed negative correlation or no correlation with spontaneous retrobulbar pain,painful eye movement,redness of eyelids and abnormal IOP.Even if 1 ≤ CAS＜3,ophthalmic symptoms and signs also showed a positive responses to iv MP in patients with moderate to severe GO as long as prolonged T2RTs occurred at least in one of EOMs.The maximal T2RTs,maximal areas,mean T2RTs and mean areas of EOMs were significantly decreased at the end of iv MP treatment,meanwhile ophthalmic symptoms and signs were obviously improved.Conclusions Prolonged T2RTs,especially accompanied by enlarged area on the identical EOMs is an remarkable symbols of acute inflamed muscles.The findings suggested that the use of prolonged T2RTs can improve diagnostic sensitivity and predict the curative effect of iv MP in patients with moderate to severe GO and CAS ≥ 1.