Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Brain Neoplasms ; drug therapy ; secondary ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Humans ; Quinazolines ; pharmacology ; therapeutic use ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism

Objective To evaluate the effects of ozoned water on the synovial inflammation in rabbits with knee osteoarthritis.Methods Thirty-two rabbits were randomly and evenly divided into four groups by random number method.All the rabbits were made into osteoarthritis models except those in groups A and D.After the osteoarthritis models were made successfully,rabbits in groups C and D received intra-articular injection of ozoned water of 20 μg/ml (2 ml)once a week for three weeks,and the other two groups did not.The morphology of synovium was observed and the expres-sion levels of IL-6 and TNF-α in the synovium were compared among the four groups.Results In group A,there was no hyperemia,edema or cell hyperplasia in the synovium,and the synovium re-mained normol tissue structure.In group B,the synovial structure was damaged,with serious cell hyperplasia,masses of inflammatory cells invading,vascular proliferation and hyperemia,and signifi-cantly increased synovium thickness compared with the normal.In group C,synovial hyperemia and edema were improved,the inflammatory cells reduced,and the synovium thickness was thinner than that in group B.And the group D had no synovitis phenomenon.Compared with group A,the expres-sions of IL-6 and TNF-αwere slightly higher in group D,and they were significantly increased (P <0.05)in the other two groups.Compared with group B,the IL-6 and TNF-α contents of synovium were reduced (P < 0.05)in group C.Conclusion Injecting 2 ml ozoned water of 20 μg/ml into artic-ular cavity can significantly improve synovial inflammation and reduce the expression of IL-6 and TNF-αin the synovium,which does no damage normal synovium.

Objective To explore the effect of levetiracetam(LEV)on cognitive function and quality of life of patients with partial seizure.Methods There were two phase for this study.Phase one was a randomized,double-blind,placebo-controlled study for 16 weeks.Phase two was an open label period for 24 weeks.Medicated patients underwent test on the cognitive function and Health Related Quality of Life (HRQOL)at the baseline respectively at the end of 16 weeks and at the end of 40 weeks.Results(1) Compared the result at the end of 16 weeks with the baseline,LEV group used less time to do Wisconsin card sorting test(WCST,reduce scores -110.3),delayed logical memory were improved(reduce scores 3.4,P

OBJECTIVETo investigate the dynamic changes that occur in T cell subsets, particularly involving the surface expression of programmed death 1 (PD-1), in response to pegylated (Peg)-interferon (IFN) a-2a therapy in patients with chronic hepatitis C virus (HCV) infection.

METHODSTwenty-five patients with HCV genotype 1b chronic infection and 10 healthy controls were enrolled in the study. All the HCV patients received combination antiviral therapy of Peg-IFNa-2a (180 mug/week) plus ribavirin. At treatment weeks 0 (baseline), 4, 12, 24 and 48, the level of PD-1 protein expression on the surface of total peripheral CD8+ and CD4+ T cells was determined by flow cytometry and the level of PD-1 mRNA expression in peripheral blood mononuclear cells (PBMCs) was determined by reverse transcription-polymerase chain reaction. Independent student's t-test were used to compare mean values between the two groups, repeat measure variance analysis was used to compare mean values among multiple groups, and Pearson's correlation coefficient was used to assess correlation significance.

RESULTSOver the course of antiviral therapy, the proportions of CD4+ T cells and CD8+ T cells, as well as the CD4+/CD8+ ratio, increased (F = 81.23, 39.28, and 7.01 respectively; all P less than 0.01). In contrast, the PD-1 protein expression frequency on CD4+ T cells and CD8+ T cells significantly declined (F = 100.11 and 158.40 respectively; all P less than 0.01). The PD-1-mRNA expression level in PBMCs was: 1.40+/-0.26 at baseline, 1.30+/-0.27 at week-4, 1.14+/-0.18 at week-12, 1.06+/-0.26 at week-24, and 0.83+/-0.25 at week-48 (F = 20.09; P less than 0.01). A positive correlation existed between the PD-1 protein expression frequencies on CD4+ T cells and CD8+ T cells and the HCV RNA load detected at baseline (r = 0.82 and 0.75 respectively; all P less than 0.01).

CONCLUSIONThe ability of Peg-IFN-a-2a-based antiviral therapy to suppress HCV replication may involve reduction of PD-1 protein expression on the surface of CD8+ T cells and CD4+ T cells.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; CD4-CD8 Ratio ; Case-Control Studies ; Female ; Hepatitis C, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Programmed Cell Death 1 Receptor ; metabolism ; Recombinant Proteins ; therapeutic use ; Ribavirin ; therapeutic use ; T-Lymphocyte Subsets ; metabolism ; Young Adult

Objective To explore the effects of lamotrigine on the cognitive function and the quality of life in epilepsy patients.Methods This was a prospective study and 91 newly diagnosed epilepsy patients were enrolled.The neuropsychological tests score and the quality of life in epilepsy inventory(QOLIE-31) were obtained before and after the treatment with lamotrigine.A battery of neuropsychological tests comprised the auditory verbal learning test(AVLT), the logical memory test(LMT), the digital symbol test(DST), the stroop color word test(SCWT), the trail making test(TMT), the verbal fluency test(VFT), the WAIS block design test(WBDT), the WAIS digital span test(WDST)and the Boston naming test(BNT). Results The repeated assessments in the patients taking lamotrigine were associated with significant improvements in many domains.The greatest changes were observed in the immediate and delayed recall of AVLT, DST, the time consuming of SCWT card C and TMT test A and B, the immediate and delayed recall of LMT, VFT, WBDT and BNT.For the quality of life, significant improvements were recorded in the fields of the seizure worry(38.81?16.06 vs 45.68?15.18), the overall quality of life(59.12?13.50 vs 64.99?13.33), the social function(64.59?25.14 vs 69.41?22.70)and the self-health evaluation (71.18?13.73 vs 76.75?11.30).Conclusion Improvements of the cognitive function and the quality of life can be observed in the initial period of medication with lamotrigine in epilepsy patients.

Objective To test the reliability and validity of the Chinese version of Profile of Fatigue Scale (PROF)among the patients with Sjogren′s syndrome. Methods The English version of PROF was translated into Chinese according to Brislin Translation Model. 107 patients with Sjogren′s syndrome were investigated to test its reliability and validity. Results The Cronbachαcoefficient , Guttman split-half coefficient, test-retest reliability and content validity of Chinese version of PROF were 0.976, 0.953, 0.872 and 0.875. Two factors were extracted by factor analysis and the cumulated variance was 80.75%. The structure of PROF was in line with the original scale. The criterion-related validity was 0.621 measured by comparing with visual analog scale of fatigue, and was -0.707 measured by comparing with vitality domain in the MOS item Short From Health Survey (SF-36). Conclusions The Chinese version of PROF has been proved to be reliable and valid. It can be used to measure the fatigue in Chinese patients with Sjogren′s syndrome.

Purpose To investigate the expression of Cav-1 in gastric cancer ( GC) cell lines and GC samples, and to analyze the pos-sible effect of gene methylation in expression of Cav-1. Methods Methylation specific PCR ( MSP) method was applied to examine the CpG methylation of the Cav-1 promoter in GC cell lines (AGS, MKN45, BGC-823) and 104 samples of GC and corresponding ad-jacent tissues. RT-PCR method was applied to examine the mRNA expression in GC cell lines. IHC were applied to examine the pro-tein expression of Cav-1 in the GC tissues. Results The expression level of Cav-1 mRNA was obviously increased after treated with 5-aza-2’-deoxycytidine (5-Aza-Dc, a demethylation agent) in AGS cell line. We detected the positive expression of Cav-1 gene in MKN45 and BGC-823 cell lines before and after treated with 5-Aza-Dc. The level of Cav-1 mRNA expression was no any change in AGS, MKN45 and BGC-823 cell lines treated with trichostatin A (TSA). MSP results showed that it can be amplified methylated bands in AGS cell line, and the methylated bands disappeared after treated with 5-Aza-Dc. MKN45 and BGC-823 cell lines were no any methylated bands amplified before and after treatment. The methylation frequency of Cav-1 gene was 29. 8% (31/104), which was significantly higher than that in adjacent tissues (P=0. 000). Furthermore, Cav-1 gene hypermethylation status was correlated with lymph node metastasis and family history of upper gastrointestinal cancers ( UGIC) , but not with pathological grade and clinical stage (P>0. 05). The positive frequency of Cav-1 expression was 51. 9%(54/104) in GC, which was significantly lower than that in adja-cent tissues (P=0. 000). The expression of Cav-1 was correlated with the frequency of gene methylation in GC tissues (P=0. 000). Conclusion The expression of Cav-1 was reduced in GC tissues and the gene hyermethylation may be one of the mechanisms causing Cav-1 gene silencing.

Background and purpose: As one of the important epigenetic phenomena, DNA methylation plays an important regulatory function for the expression of genes. Study shows that abnormal changes of DNA methy-lation patterns of normal tumor cell genome leads to dysfunction of cancer related gene, and this may be associated with tumor occurrence and development. The study investigated the promoter methylation and expression of caveolin-1 (CAV-1) gene in esophageal squamous cell carcinoma (ESCC), and to elucidate its role in ESCC. Methods:We used MSP approach, RT-PCR, and immunohistochemistry method respectively to examine the methylation status of the 5’CpG island of CAV-1 gene and its expression at mRNA and protein levels in tumors and corresponding normal tissues. Results: CAV-1 mRNA expression in tumor tissues (0.86±0.56) was signiifcantly higher than that in corresponding normal tissues (0.40±0.36, P<0.05). The mRNA expression of CAV-1 was correlated with status of lymphatic metastasis and TNM stage of ESCC patients (P<0.05). The protein expression of CAV-1 in tumor specimens (66.7%, 34/51) was signiifcantly higher than that in corresponding normal tissues (15.7%, 8/51, P<0.01). The protein expression of CAV-1 was signiifcantly associated with lymphatic metastasis of ESCC (P<0.05), however, it was not associated with differen-tiation and TNM stage (P>0.05). The promoter methylation frequency of CAV-1 in tumor specimens was 2.0%(1/51), and the methylation phenomenon has not been found in corresponding normal tissues. The promoter methylation fre-quency of CAV-1 in tumor specimens showed no signiifcant difference compared with the corresponding normal tissues (P>0.05). Conclusion:The mRNA and protein expression of CAV-1 in tumor specimens was signiifcantly higher than that in corresponding normal tissues. Aberrant high expression of CAV-1 has played a certain role in promoting tumori-genesis and lymph node metastasis. The expression both in ESCC and corresponding normal tissues has no correlation with the promoter methylation status.

Objective To evaluate the safety and efficacy of Zonisamide(ZNS) as adjunctive therapy in patients with refractory partial seizures receiving other antiepileptic drags (AEDs).Methods This was a randomized,double-blind,placebo-controlled study conducted at multi-centers.All 240 subjects were randomized to either the ZNS group or the placebo group in a 1:1 ratio.The double-blind treatment phase included a titration phase during which zonisamide dose inereased from 100 mg/day to 300 mg/day over 4 weeks and then a 12-week fixed-dose phase.The primary efficacy endpoint was,the median % reduction from baseline in all pattial seizure frequency(CPS+SPS+SGS)during the fixed-dose phase.The important secondaw endpoint wag the responder rate.Safety profiles and tolerance were also evaluated.Results The FAS analysis showed the median reduction from baseline in the ZNS group was greater than in the placebo group(48.4%vs 26.6%),the difference was significant for ZNS compared with placebo(F=4.904,P=0.028);The responder rates for all partial seizures(48.6%vs34.9%,X2=4.046,P=0.044)and for complex seizures(52.2% vs 33.3%,X2=5.607,P=0.018)were significantly higber in the ZNS group than in the placebo group in the FAS population.The overall adverse events(AEs)profile was comparable between the two groups.The most frequent AEs considered to be related to zonisamide by the investigator were headache,dizziness,somnolence,anorexia,nausea,etc.Conclusions ZNS is superior to placebo in reducing the frequency of partial seizures and well-tolerated.ZNS could be a choice of adjunctive therapy in patients with refractory partial seizures.