OBJECTIVES: To investigate CEACAM6 expression in nasopharyngeal carcinoma (NPC) and its regulatory effects on tumor cell proliferation, migration, and epithelial-mesenchymal transition (EMT). METHODS: CEACAM6 expression in NPC was analyzed using GEO datasets and validated by immunohistochemistry in NPC tissues and by Western blotting and RT-qPCR in NPC cell lines (HNE1, C666-1, HK1, 5-8F and CNE2Z) and normal nasopharyngeal epithelial NP69 cells. In the NPC cell lines, the effects of lentivirus-mediated CEACAM6 overexpression and knockdown on cell proliferation, migration, invasion and cytoskeletal structures were evaluated using CCK-8 assay, Edu staining, wound healing assay, Transwell assay, and phalloidin staining. Western blotting was performed to determine the expressions of EMT-related proteins (FN1, ITGA5, ITGB1, E-cadherin, N-cadherin and vimentin) in the NPC cells and the effect of FN1 overexpression on ITGA5 and ITGB1 protein expressions. RESULTS: Analysis of the data from the GEO datasets suggested that CEACAM6 was significantly downregulated in NPC, which was associated with poor patient prognosis. Immunohistochemistry also showed low expressions of CEACAM6 in clinical NPC tissues (P<0.05). In NPC cells, CEACAM6 overexpression significantly suppressed cell proliferation, migration and invasion and reduced the fluorescence intensity of actin. CEACAM6 overexpression also resulted in significant downregulation of FN1, ITGA5, ITGB1, N-cadherin and vimentin expressions and upregulation of E-cadherin expression, and FN1 overexpression obviously attenuated the inhibitory effect of CEACAM6 overexpression on ITGA5 and ITGB1 expressions. CONCLUSIONS CEACAM6 inhibits NPC cell migration and invasion by inhibiting EMT via regulating FN1, ITGA5 and ITGB1 expressions.
Humans ; Epithelial-Mesenchymal Transition ; Cell Movement ; Cell Proliferation ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms/metabolism* ; Cell Line, Tumor ; Cell Adhesion Molecules/genetics* ; Antigens, CD/metabolism* ; GPI-Linked Proteins ; Integrin alpha5/metabolism* ; Integrin beta1/metabolism* ; Cadherins/metabolism* ; Fibronectins ; Integrins

BACKGROUND: Biological agents, such as tumor necrosis factor inhibitors (TNFi), have been widely used in rheumatoid arthritis (RA) patients and greatly improved goal achievement. The aim of this study was to investigate whether conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs) combination was better in reducing relapse than methotrexate (MTX) monotherapy, and more cost-effective than continuing TNFi plus MTX in RA patients who achieved low disease activity (LDA) with TNFi and MTX therapy. METHODS: RA patients who failed to csDMARDs received an induction therapy of MTX plus TNFi for maximally 12 weeks. Those achieving LDA in 12 weeks were randomly assigned at a 1:1:1 ratio into three groups: (A) adding hydroxychloroquine and sulfasalazine for the first 12 weeks and then discontinuing TNFi for the following 48 weeks; (B) maintaining TNFi and MTX for 60 weeks; and (C) maintaining TNFi and MTX for the first 12 weeks and then discontinuing TNFi for the following 48 weeks. The primary outcome was relapse. RESULTS: A total of 117 patients were enrolled for induction therapy and 67 patients who achieved LDA within 12 weeks were randomized, with 24, 21, and 22 patients in groups A, B, and C, respectively. The relapse rates of groups A and B during the entire 60 weeks were comparable [10/22 (45.5%) vs. 7/20 (35.0%), χ2 = 0.475, P = 0.491], however, significantly lower than that of group C [10/22 (45.5%) vs. 17/20 (85.0%), χ2 = 5.517, P = 0.019; 7/20 (35.0%) vs. 17/20 (85.0%), χ2 = 11.035, P = 0.004, respectively]. Taking RMB 100,000 Yuan as the threshold of willingness to pay, compared to MTX monotherapy (group C), both TNFi maintenance and triple csDMARDs therapies were cost-effective, but triple csDMARDs therapy was better. CONCLUSION: For RA patients who have achieved LDA with TNFi and MTX, csDMARDs triple therapy was a cost-effective option in favor of reducing relapse. TRIAL REGISTRATION ClinicalTrials.gov, NCT02320630.
Humans ; Cost-Benefit Analysis ; Drug Therapy, Combination ; Treatment Outcome ; Arthritis, Rheumatoid/drug therapy* ; Antirheumatic Agents/therapeutic use* ; Methotrexate/therapeutic use* ; Recurrence

Coding system is the basis of delicacy and informatization management of medical supplies, and is the link between enterprise production and clinical use. Rational construction of coding system can realize dynamic analysis of big data, guide clinical use, improve the use efficiency, and form a traceability system. Coordinating with medical security standardization work guidance, the authors analyzed the characteristics of current classification, and constructed a new medical supplies coding system by combining the clinical characteristics, price supervision, level of risk, properties of medical charge and insurance, production date and traceability, etc. This new medical supply coding system could contribute to analyzing the usage structure and strengthening price supervision, medical supplies classification and traceability management, which could be appropriate for the monitoring of the whole supply chain, from production, logistics, procurement, supplies usage, charge, medical insurance to traceability management.

Objective To establish a prokaryotic expression system of interstitial lung disease associated autoantigen human bactericidal/permeability-increasing fold-containing B1 (BPIFB1), providing tools for the study on its function in immune responese. Methods The coding region of BPIFB1 gene was amplified with specific primers from recombinant pGEM-C20ORF114 plasmid and cloned into the pET28a-MBP-His and pGEX-5X-1 vectors. The recombinant pET-BPIFB1-MBP-His and pGEX-BPIFB1-GST plasmids were transfected into Top10 cells. The positive clones were selected and sequenced. The correct clones of pET-BPIFB1-MBP-His and pGEX-BPIFB1-GST were transfected into prokaryotic expression strain Rosetta (DE3) and induced by Isopropyl β-D-Thiogalactoside (IPTG). The expression of recombinant BPIFB1 fusion protein was analyzed by SDS-PAGE and Western blotting, and purified by urea modified and renaturation and affinity chromatography of nickel NTA-resin. Results The polymerase chain reaction (PCR) produced specific product with the molecular weight equivalent to that of BPIFB1. The recombinant pET-BPIFB1-MBP-His and pGEX-BPIFB1-GST plasmids were cloned by double restriction enzyme digestion and ligation and confirmed by sequencing. The SDS-PAGE result showed that both BPIFB1-MBP and BPIFB1-GST fusion proteins were mainly expressed in the form of inclusion bodies. The Western blotting result revealed that the recombinant BPIFB1-MBP-His protein could be recognized by Anti-6 ×His antibody. The purified soluble BPIFB1-MBP fusion protein was obtained by urea denaturation, affinity chromatography of nickel NTA-resin and then renaturation after purification. Conclusion The BPIFB1 prokaryotic expression system is established by construct recombinant plasmid pET-BPIFB1-MBP-His, and an approach of renaturation after nickel resin affinity purification in denatured condition.

Objective To investigate the expression of histone deacetylase 8 (HDAC8) in the labial gland of patients with Sj?gren's syndrome (SS), and whether it is related to the abnormal activation of B cells of SS patients. Methods The expression of HDAC8 was detected by immunohistochemistry in the labial glands from SS patients (n=10) and non-SS patients (n=4). The subtype of lymphocytes which expressed HDAC8 was determined by double staining immunofluorescence. Person correlation analyses were performed to evaluate the relation between the proportion of HDAC8 positive cells and the level of immunoglobulins in SS patients. Results The expression of HDAC8 in the labial gland from SS patients was significantly higher than that in the non-SS patients.The proportion of HDAC8 positive cells in lymphocytic foci was higher than that in scattered interstitial lymphocytes [(0.76±0.05) vs (0.40±0.03), t=18.5, P<0.01]. IF showed that HDAC8 was expressed in CD20+B cells, and CD138+plasma cells in labial gland from SS group,but not CD4+T cells. The ratio of HDAC8+cell in infiltrating lymphocytic loci was not related to the level of IgG or IgA of SS patients. Conclusion HDAC8 is expressed in CD20+B cells and CD138+plasma cells in labial gland from SS patients. These data suggests that HDAC8 may be involved in abnormally activated B cells and plasma cells in SS patients.

Objective To explore the characteristics and survival of connective tissue disease (CTD) patients with both of pulmonary arterial hypertension (PAH) arnd interstitial lung disease (ILD),and to compare with CTD patients with isolated PAH.Methods All adult CTD patients who visited one of the three referral centers in China with a diagnosis of PAH confirmed by right heart catheterization from July 2006 to May 2011 were enrolled.They were then divided into two groups (ILD with and without-ILD group) based on chest CT and then the comparison of baseline characteristics and survival at the endpoint of follow up were made between the two groups.T test,Mann-Whitney U test,x2 test,Kaplan-Meier survival analysis and Cox regression analysis were used for statistical analyses.Results One hundred and twenty-six patients were recruited into the study.Patients with ILD (n=27) were older than those without ILD (n=99).Lung function results including FVC [(75±18)％ vs (83±13)％,t=2.212,P=0.037] and DLCO [(54±22)％ vs (68±20)％,t=2.392,P=0.019] in ILD group were significantly wose than those without-ILD group.Although some important hemodynamic parameters such as mean pulmonary arterial pressure and pulmonary vascular resistance were better in the ILD group than the without-ILD group,Kaplan-Meier analysis showed that the short term survival of ILD group was significantly worse than that of the without-ILD group (72.7％ versus 94.7％ at 1 year and 63.6％ versus 81.1％ at 3 year,P=0.047).In ILD group,Cox regression analysis showed that SvO2 was the only independent factor for the short term survival [HR=0.19,95％CI (0.04,0.83),P=0.027],and Kaplan-Meier analysis showed patients with SvO2＜60％ had significantly lower short term survival than patients with SvO2 ≥60％ (1 and 2 year survival were 60.0％ and 40.0％ versus 92.9％ and 77.4％ respectively,P=0.002).Conclusion Patients with both PAH and ILD is a special subtype in CTD.Although with the superiority of hemodynamics,these patients have significantly worse survival than CTD patients with isolated PAH.Low SvO2 is the independent risk factor for the short term mortality in patients of CTD complicated by both PAH and ILD.More attention should be paid to these patients and the management strategy should be investigated further.

Objective To understand the mental health status between left‐behind children and un‐left‐behind children during primary school period in Yongchuan district of Chongqing city ,and to provide the theoretical basis for the study of the left‐behind children children′s mental problems and formulating the strategy and measures for preventing their psychological problems .Methods The Mental Health Rate Scale for Pupil (M HRSP) was used to evaluate the mental health condition in 4 987 primary school pu‐pils from 4 primary schools in Yongcuan discrict ,including 3 482 un‐left‐behind children(69 .82% ) and 1 505 left‐behind children (30 .18% ) .Results Totally 4 753 effective questionnaires were taken back with the effective rate of 95 .31% .The detection rate of total score deviation in left‐behind children was 13 .98% ,which was significantly higher than in un‐left‐behind children(6 .33% ) . The deviation detection rate of learning disabilities in left‐behind children was 14 .90% ,emotional disorder was 16 .81% ,character flaw was 12 .36% ,social adjustment disorder was 13 .91% ,moral defect was 14 .27% ,bad habits was 15 .61% ,behavior disorder rate was 16 .53% and the special obstacles was 11 .72% .Compared with un‐left‐behind children ,excluding the character flaw ,social adjustment and moral defect ,the deviation detection rate and scores of other 5 items in left‐behind children were higher than those in un‐left‐behind children .The scores of learning disabilities ,emotional disorder and bad habits in male pupils were higher than those in female pupils with statistical difference(P< 0 .05) .The scores of learning disabilities ,emotional disorder ,character flaw and behavior disorder in the high grade pupils were higher than those in the low grade pupils ,the differences were statistically sig‐nificant(P<0 .05) .Conclusion The mental health level of rural left‐behind children in Yongchuan district is low ,the problems are more serious compared with un‐left‐behind children .In carrying out the mental health education work in primary school pupils ,es‐pecially the mental health education should be paid attention to rural left‐behind children ,moreover the mental health education should be carried out aiming at different ages and different genders .

Objective:To verify the fracture risk assessment tool ( FRAX) to estimate the probability of osteoporotic fracture in patients with rheumatoid arthritis ( RA ) with or without bone mineral density (BMD), and identify associated risk factors of osteoporosis .Methods: In the study, 200 patients with rheumatoid arthritis aged more than 40 years in Peking University First Hospital from Dec .2009 to Dec. 2012 were recruited.Clinical information was obtained from a questionnaire of their case history and medical records.FRAX tool was administered.Their lumber spine and left femoral BMD were determined by dual energy X ray absorptiometry.The gender, age, disease duration, menopause status, body mass index ( BMI) and accumulative dose of glucocorticoid were obtained in retrospect .Correlation analysis was conducted between the BMD and clinical information .Results:The study population ( female, 77.5%) had a mean age of 59.4 years, in which 10 (13%) patients showed a normal BMD, 67 (87%) were osteopenia or osteoporosis , while 32 patients (16%) had fragile fracture.Compared with the patients with normal BMD, the subjects with low BMD had significantly older age , longer period for corticoids usage , higher day dose and accumulated dose of corticoids .The 10-year fracture risk of sustai-ning major osteoporotic fractures and hip fracture was higher .No significant difference was observed be-tween the 10-year fracture risks calculated with BMD and without BMD .The values of the different area under the receiver operating characteristic ( ROC) curve ( AUC) for major and hip fractures calculated in three ways:without BMD, with the femoral neck BMD, and with T-score.The best result was for FRAX tool for hip fracture with the T-score ( AUC 0 .899 ) .A stepwise multivariate linear regression model was constructed to explore the relationship between the different clinical factors studied and a low BMD . Three statistically significant variables for lumber BMD were pain on visual assessment scale ( VAS ) (P=0.02), fracture history (P=0.003) and a higher steroid accumulated dose (P=0.008).Three statistically significant variables for left hip BMD were age (P<0.001), fracture history (P=0.05) and lower BMI ( P=0.03) .Conclusion:Low BMD is a common complication in RA patients .Risk factors for major fracture and hip fracture are increased .There is a positive correlation between FRAX calculated with and without BMD or T score .FRAX with the femoral neck T score or BMD presents a discriminatory capacity better than FRAX without BMD , according to the AUC ROC .

Objective To analyze the nuclear factor-kappa-B activator 1 (Act1) expression in synovial tissues and peripheral blood lymphocytes in patients with rheumatoid arthritis (RA),and explore its possible regulatory mechanisms.Methods The Act1 expression of synovial tissue from 10 RA patients and 5 osteoarthritis patients were analyzed by immunohistochemical staining,and double immunofluorescence staining was used to identify the Act1-expressing cells.The peripheral blood of 47 RA patients and 22 healthy controls (HC) were obtained.The Act1 expression in different lymphocyte subpopulation from peripheral blood was determined by flow cytometry and Act1-mRNA expression was detected by real-time quantitative polymerase chain reaction (RT-PCR).The data was analyzed by Mann-Whitney U test,two independent sample t test and Pearson correlation analysis.Results Remarkable infiltration of inflammatory cells and expression of Act1 in synovium from RA patients was observed.The Act1 was expressed in T lymphocytes,in contrast to the few expression in B lymphocytes.Compared to HC,the expression of Act1-mRNA was lower in peripheral CD4+ T cells and CD8+ T cells [CD4+ T cells:133 (69,380) vs 158 (117,246),U=36,P＞0.05; CD8+ T cells:127(63,363) vs 129(81,222),U=35,P＞0.05],however,higher in CD20+ B cells in RA patients [62(34,81) vs 52(33,105),U =36,P ＞0.05].There was no significant difference of Act1-mRNA levels in peripheral blood mononuclear cells between HC and RA patients (0.71 ±0.32 vs 0.79±0.21,t=1.717,P=0.091).However,a tendency of down-regulation of Act1 expression was noticed in RA patients than HC.No significant correlation was found between Act1-mRNA level and RA disease activity index.Conclusion Our data suggest that Act1 may be involved in the pathogenesis of RA,however,we could not yet found its association with RA disease activity.

Objective To investigate the outcomes of patients with rheumatoid arthritis (RA) treated by different combination of synthetic disease modifying antirheumatic drugs (DMARDs) under the guidance of treat-to-target strategy.Methods Forty-two RA patients with high disease activity were enrolled into this randomized,open-label and prospective study.It was comprised of a maximal 36-week induction phase and then followed by a maintenance phase up to 84 weeks.Combination of synthetic DMARDs was initiated in the induction phase,with or without low dose glucocorticoids (GCs) during the first 12 weeks.Patients who achieved low disease activity (LDA) were randomized into two maintenance groups.An increase of DAS28 by 0.6 was defined as relapse.The patients achieved LDA in the induction phase,relapsed during maintenance phase and possible relevant risk factors were analyzed.Results Twenty-seven (64％) patients achieved LDA during the induction phase.More non-smoking patients achieved LDA than those smoked [85％ (11/13) vs 55％(16/29),P＜0.05].During the maintenance phase,14 (61％) out of 27 patients relapsed.Patients taking GCs during the first 12 weeks had a significantly higher relapse rate compared to those without GC (83％ vs 36％,P=0.021).Patients who entered maintenance phase at week 12 had a significantly higher tendency to relapse compared to those who entered the maintenance phase at week 24 [75％(9/12) vs 33％(3/9),P=0.026].Conclusion Smoking seems to be a risk factor for RA patients who fail to reach LDA.Low dose GCs as a bridge therapy may require a longer duration.High relapse rates in both the maintenance groups indicat that a longer tight induction phase may be appropriate before downstairs therapy.