Adolescent ; Adult ; Aged ; Facial Nerve ; anatomy & histology ; pathology ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Parotid Neoplasms ; surgery ; Young Adult

Objective To study the effects of citrullinated vimentin (cVim) on the maturation and immunologic function of dendritic cells (DCs) from rheumatoid arthritis (RA) peripheral blood.Methods In the present study,mononuclear cells were isolated from the peripheral blood of patients with RA and cultivated in media containing GM-CSF and IL-4 to generate immature DCs (imDCs).The imDCs generated were stimulated with citrullinated vimentin and vimentin.LPS was used as the positive control and PBS was used as the negative control.The expression of surface molecules on the DCs,such as CD14,CD80,CD83,CD86,MHC Ⅰ and MHC Ⅱ were analyzed with FACS.The capability of the stimulatory activity of the DCs on allogeneic T cells in mixed reaction was tested by MTS.t-test was used for statistical analysis.Results Compared to untreated DCs,DCs treated with LPS increased the expression levels of MHC Ⅱ,CD80,CD83 and CD86 (1.07±0.14,1.25±0.13,1.90±1.08,2.44±0.65,P＜0.05),while cVim increased the expression levels of MHC Ⅱ ( 1.18±0.09,P＜0.05) and CD83 ( 1.97±0.99,P＜0.01 ),and Vim decreased the expression levels of CD80 (0.82±0.18,P＜0.01 ).It was demonstrated that the expression levels of MHC Ⅱ on DCs pulsed with cVim were significantly higher than that of the DCs with LPS,but the expression levels of CD80 and CD86 were not significantly different.The expression levels of MHC Ⅱ and CD83 on DCs pulsed with cVim were significantly higher than that of the DCs with Vim.The mixed lymphocyte reaction showed that the DCs induced by LPS and cVim trigerred the proli-feration of allogenic T cells obviously.Conclusion This result suggests that cVim could promote the phenotypic maturation of DCs and increase the expression of costimulatory molecules.

China ; Congresses as Topic ; Fatty Liver ; Humans

OBJECTIVETo investigate the methylation status of zonula occludens-1 (ZO-1) gene promoter and its clinical significance in children with stage IV non-Hodgkin lymphoma (NHL) and to provide a basis for further etiological study and early diagnosis of this disease.

METHODSFifty-five children with a confirmed diagnosis of stage IV NHL (40 cases of T-NHL and 15 cases of B-NHL) were selected as the case group, and 20 children with diseases other than hematologic malignancies were selected as the control group. Bone marrow samples were collected from these subjects. Methylation-specific PCR (MS-PCR) was applied to evaluate the methylation status of ZO-1 gene promoter, and the integrated optical density (IOD) was determined. RT-PCR was used to measure the mRNA expression of ZO-1.

RESULTSMS-PCR showed that the methylated bands of ZO-1 gene promoter were found in 39 (70.9%) of 55 patients in the case group before treatment, while no ZO-1 gene promoter methylation was detected in the control group. With close tracking of 47 cases in the study group, consisting of 32 cases of T-NHL and 15 cases of B-NHL, the rates of ZO-1 gene promoter methylation prior to treatment were 72% and 67%, respectively, (P>0.572). The cases of T-NHL and B-NHL showed no significant changes in methylation rate in the early and middle phases of chemotherapy (P>0.05), but they showed significant changes in methylation rate in the late phase of chemotherapy (P<0.05). RT-PCR showed that the NHL cases carrying methylated ZO-1 gene had no mRNA expression of ZO-1, while all children in the control group had mRNA expression of ZO-1. There was no linear relationship between the total number of peripheral blood leukocytes and ZO-1 gene IOD (r=0.093, P=0.575); a positive correlation was found between the number of malignant cells in bone marrow and ZO-1 gene IOD (r=0.669, P<0.001).

CONCLUSIONSZO-1 gene shows a hypermethylation status in children with NHL, and the methylation level is positively correlated with the number of malignant cells in bone marrow. ZO-1 may be used as a novel molecular marker in early diagnosis, outcome assessment, prognostic evaluation, and detection of minimal residual disease.

Adolescent ; Child ; Child, Preschool ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma, Non-Hodgkin ; genetics ; Male ; Promoter Regions, Genetic ; Zonula Occludens-1 Protein ; genetics

Objective To investigate the relationship between gene polymorphism of transcripion factor 7-like 2 (TCF7L2) at positions rs290487, rs11196205, rs11196218 and gestational diabetes mellitus (GDM) in Chinese women.Methods In 1140 unrelated pregnant Northern Chinese women (335 women with GDM, 158 gestational cases with impaired glucose tolerance and 647 pregnant non-diabetic controls) ,three single nucleotide polymorphisms (rs290487, rs11196205, and rs11196218) in the TCF7L2 gene were genotyped using ligase detection reaction (LDR).In the present study, cases with GDM and impaired glucose tolerance (IGT) were indistinguishable clinically and biochemically, and were combined into case group.Results The frequency of C allele of rs290487 was 41.6% in case group, being significantly higher than that in control group (36.3%, P=0.012).There was significant difference in the frequency of CC genotype between case group and control group (18.7% vs 14.0%, P=0.033).Compared with T allele carriers, CC genotype carriers had a 1.418-fold increased risk of GDM (95% CI 1.028-1.955).After adjusting for age, body mass index, family history of diabetes,systolic blood pressure,and diastolic blood pressure, pregnant women with CC genotype carriers of rs290487 were more prone to hyperglycemia compared with the T allele carriers (OR 1.518, 95% CI 1.064-2.166).Conclusions The TCF7L2 rs290487 variant may contribute to the genetic predisposition to GDM.CC genotype is likely to be associated with an increased risk of GDM in the pregnant Chinese women.

Objective:To prove the effect of PADI-4 gene in the development of rheumatoid arthritis.Methods:Four SiRNA sequences were designed for PADI-4 gene,and the SiRNAs were cloned into blank pSiRNA-hH1neo G2 vectors.The vectors were transformed into GT116 E.coli competent cells.By white-blue selection system,the right vectors were gotten.After transfection into HL-60 cells,the cells were collected on 3,5,7,10 and 14 day,the levels of PADI-4 mRNA were detected by Real-time PCR.Results:Digestion by Acc 65Ⅰand Hind Ⅲ,the recombinant expressive vector of RNA interference was obtained successfully.The PADI-4 mRNA generated by the cells transfected with the vector of SiRNAs were reduced,and the level was not change in normal cells.Conclusion:The recombinant expressive vector of RNA interference is obtained successfully and the recombinant expressive vector can affect expression of PADI-4 gene in HL-60 cells.

Objective To study the injury and protective action of drugs on neurons, the model of glutamate excitotoxicity on primary cultured hippocampual neurons from new born rats were( set up. Methods)By use of trypan blue dye staning and testing the lactate dehydrogenase leakage from cultured neurons, to investigate the neuron survival rate. Results We found the injury of neurons was related with the concentration of glutamate. NMDAR non-competitive antagonist —MK-801 could protect the glutamate excitotoxic damage on neurons. Conclusion The glutamate results in neuron injury through NMDAR; the model of neuron culture was sufficient for glutamate-induced excitotoxicity.

AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Objective Triple negative breast cancer(TNBC), a special breast cancer subtype, is lack of effective target therapy.The article aimed to investigate the role of lysophosphatidic acid (LPA) and Hippo Yes-associated protein (Hippo-YAP) signaling pathway in TNBC invasion and metastasis and the mechanisms.Methods The specific small interfering RNA (siRNA) of YAP was synthetized in vitro, and was transfected into MDA-MB-231 cells using liposome transfection.The experiment was divided into YAP-siRNA group, positive control group and blank control group.Each group is provided with 2 parallel holes.Evaluation was made on the effects of each group on Hippo-YAP, the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group, YAP content, the capacity of invasion and metastasis after transfection ([0.035±0.005], [2.200±1.000], [3.500±0.800]) significantly decreased compared with positive control group([0.343±0.012], [27.600±5.100], [22.300±5.000]) and blank control group([0.384±0.017], [26.500±4.800], [22.350±6.000]) (P<0.05).YAP expression levels at 60 min, 120 min, and 240 min in experiment group significantly decreased compared with positive control group and blank control group (P<0.05).YAP relative expression levels of 10, 20, 50 μmol/Lwere significantly lower than those of positive control group and blank control group (P<0.05).After respective interference of C3 transferase and Y27623, significant difference was found in the pYAP mRNA contents of experiment group([0.255±0.052], [0.326±0.017]), blank control group([0.048±0.032], [0.534±0.017]) and positive control group([0.052±0.021], [0.528±0.024])(P<0.05).The expression levels of YAP mNA and AREG mNA significantly increased in experiment group([0.176±0.032], [0.263±0.008]) compared with blank control group([0.043±0.013], [0.263±0.008]) and positive control group([0.049±0.025], [0.057±0.043])(P<0.05).Conclusion LPA induces breast cancer invasion and metastasis, which is YAP-dependent, time-dependent and concentration-dependent.LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.

Objective To evaluate the feasibility of using comprehensive geriatric assessment (CGA) in estimating if standard dose treatment is fit for the elderly patients with diffuse large B cell lymphoma.Methods.Comprehensive geriatric assessments including three assessments of activity of daily living,instrumental activity of daily living and comorbidity scoring according to Cumulative Illness Rating Score for Geriatrics were adopted to assess if standard dose treatment is fit for the elderly patients in our prospective study.Thirty seven patients with diffuse large B cell lymphoma,aged ＞70 years were enrolled in the study,and grouped into fit,unfit and frail groups according to comprehensive geriatric assessment scoring and their age.The treatment protocolswere not determined by comprehensive geriatric assessment scores,but by clinical judgments made by clinicians based on their clinical experience and disease features.The clinically effective response and overall survival (OS) were analyzed in the three groups.Results According to CGA scores,patients were grouped into fit [21 cases (56.8％)],unfit [7 (18.9％)] and frail [9 (24.3％)].37 cases received 213 courses of treatment at average 5.76 courses per case.The overall response (complete / partial remission) rates were [85.7％(18/21) vs.28.6％ (2/7) vs.44.4％ (4/9),x2=9.69,P=0.008] and median survival times were (44 months vs.10 months vs.9 months;x2 =7.03,P=0.03) among fit,unfit and frail groups with statistically significant differences.Total effective rate (achieving all clinical targets) in fit group of 21 cases were 100 ％ (12/12)with receiving standard dose therapy,and 66.7％ of(6/9)with low dose therapy(P=0.06).Overall response rate(total/partial remission) [85.7％(18/21) vs.28.6％(2/7) vs.44.4％(4/9),x2=9.69,P=0.008] and median survival (44 months vs.10 months vs.9 months;x2 =7.03,P=0.03) amongfit,unfit and frail groups.In fit group,the two-year overall survival was higher in patients receiving standard dose treatment than receivingpalliativetreatment,with statistical significance [83.3 ％ (10/12) vs.33.3 ％ (3/9),P =0.032],without significant hematologic toxicity observed between the subgroups.Conclusions Comprehensive geriatric assessment can identify if elderly patients diffuse large B cell lymphoma can acquire a satisfactory curative effect from a standard dose treatment ofimmunochemotherapy.