Analysis of the effect of siRNA silencing α1 antitrypsin on rheumatoid arthritis fibroblasts / 中华风湿病学杂志

Objective To investigate the effect of siRNA silencing α1 antitrypsin (α1-AT) gene on the biological behavior of rheumatoid arthritis fibroblast-like synoviocytes (RA-SFs).Methods Primary culture of knee synovial tissue from 5 patients with rheumatoid arthritis (RA) was performed.The artificially synthesized silencing α1-AT siRNA specifically inhibits the expression of α1-AT in RA-SFs.After 24 and 36 hours of transient transfection,the inhibition efficiency was detected by Quantitative reverse transcription polymerase chain reaction (RT-qPCR),and the expression of related genes after α1-AT gene silencing was detected.Furthermore,ethyl thiazolyl tetrazolium (MTT) assay,Trans-well chamber,cell scratch and enzyme-linked immunosorbent assay (ELISA) were used to detect the effects of interfering α1-AT expression on cell proliferation,invasion and migration,and secretion of interleukin (IL)-17,Tumor necrosis factor (TNF)-α,IL-1α,IL-1β and other related inflammatory factors.At the same time,when the pathway inhibitor (ERK inhibitor,signal transducer and activator of transcription 3 (STAT3) inhibitor,NF-κB inhibitor) stimulated cells,the effect on α1-AT was changed.One-way analysis of variance was used for comparison between the two groups;further pairwise comparison using LSD-t test;The count data was checked by x2 test.Results Compared with the negative control group,the siRNA α1-AT group was transfected for 24 hours,the α1-AT mRNA level was significantly inhibited (P＜0.05),and the proliferation rate of RA-SFs was not affected (P＞0.05),and the invasion and migration ability of cells decreased significantly (P＜0.05).The secretion of IL-17 and IL-1β was slightly decreased (P＞0.05),while TNF-α and IL-1α were not affected (P＞0.05).In addition,the expression of α1-AT downstream genes Matrix metallop roteinases (MMP)-2 169,Hu-Bax1,MMP-9,Hu Bax and Hu Bcl-xl were significantly decreased (P＜0.05).Moreover,the signaling pathway inhibitors ERK inhibitor (PD98059) and NF-κB inhibitor (PDTC) had significant effects on α1-AT (P＜0.05).Conclusion The α1-AT gene silencing has potential effect on the biological behavior of RA SFs.Further study of this mechanism is helpful to provide evidence for the treatment of RA.

Analysis of the effect of siRNA silencing α1 antitrypsin on rheumatoid arthritis fibroblasts / 中华风湿病学杂志

Objective@#To investigate the effect of siRNA silencing α1 antitrypsin (α1-AT) gene on the biological behavior of rheumatoid arthritis fibroblast-like synoviocytes (RA-SFs).@*Methods@#Primary culture of knee synovial tissue from 5 patients with rheumatoid arthritis (RA) was performed. The artificially synthesized silencing α1-AT siRNA specifically inhibits the expression of α1-AT in RA-SFs. After 24 and 36 hours of transient transfection, the inhibition efficiency was detected by Quantitative reverse transcription polymerase chain reaction (RT-qPCR), and the expression of related genes after α1-AT gene silencing was detected.Furthermore, ethyl thiazolyl tetrazolium (MTT) assay, Trans-well chamber, cell scratch and enzyme-linked immunosorbent assay (ELISA) were used to detect the effects of interfering α1-AT expression on cell proliferation, invasion and migration, and secretion of interleukin (IL)-17, Tumor necrosis factor (TNF)-α, IL-1α, IL-1β and other related inflammatory factors. At the same time, when the pathway inhibitor (ERK inhibitor, signal transducer and activator of transcription 3 (STAT3) inhibitor, NF-κB inhibitor) stimulated cells, the effect on α1-AT was changed. One-way analysis of variance was used for comparison between the two groups; further pairwise comparison using LSD-t test; The count data was checked by χ2 test.@*Results@#Compared with the negative control group, the siRNA α1-AT group was transfected for 24 hours, the α1-AT mRNA level was significantly inhibited (P<0.05), and the proliferation rate of RA-SFs was not affected (P>0.05), and the invasion and migration ability of cells decreased significantly (P<0.05). The secretion of IL-17 and IL-1β was slightly decreased (P>0.05), while TNF-α and IL-1α were not affected (P>0.05). In addition, the expression of α1-AT downstream genes Matrix metallop roteinases (MMP)-2 169, Hu-Bax1, MMP-9, Hu Bax and Hu Bcl-xl were significantly decreased (P<0.05). Moreover, the signaling pathway inhibitors ERK inhibitor (PD98059) and NF-κB inhibitor (PDTC) had significant effects on α1-AT (P<0.05).@*Conclusion@#The α1-AT gene silencing has potential effect on the biological behavior of RA SFs. Further study of this mechanism is helpful to provide evidence for the treatment of RA.

Review of E2F Transcription Factor 2 Expression in Rheumatoid Arthritis Fibroblast-like Synovial Cells (RA FLs) / 医学研究杂志

Objective To study the relationship between E2F2 and the pathological features of RA and the mechanism of E2F2 expression.Methods The interference efficiency was detected by RT-PCR after RA FLs transfected with E2F2 siRNA.The cell proliferation was analyzed by MTS and the expression of numerous genes was tested by ELISA.To analyze the influence of E2F2 in the immune response by RT-PCR,RA FLs were separately treated with TNF-t and PDTC.The way of regulating E2F2 experssion was tested by Chip.Results Compared with negative control groups,the level of E2F2 mRNA were decreased in the cells transfected with E2F2 siRNA (P ＜ 0.01).Silencing E2F2 suppresses the proliferation of RA FLs (P ＜ 0.05).TNF-α regulates the expression of E2F2 in RA FLs via the NF-κB pathway.Conclusion Up-regulation of E2F2 is associated with the process of RA and TNF-α regulates the expression of E2F2 in RA FLs via the NF-κB pathway.Thus E2F2 might be a potential target for use in the development of new therapeutic approches of RA.

Preliminary build of neonatal surgical nursing quality evaluation indicator system / 中国实用护理杂志

Objective To establish a scientific and objective indicator system for nursing quality evaluation according to nursing works of neonatal surgical department. Methods To establish the indicator system based on the three- dimension quality structure. Literature retrieval, expert interview and Delphi expert enquiry were used to filter and improve indicators. Analytic hierarchy process was used to determine the index weight and check the consistency. Results The quality indicator system included three levels. There were 3 indicators in level one, 14 indicators in level two and 70 indicators in level three. Experts coefficient was 0.867, mean of importance values of two rounds were 3.83-5.00 and 4.08-5.00 , Kendall coordination coefficients were 0.292(P<0.01) and 0.301(P<0.01). Conclusions The nursing quality evaluation indicator system has high reliability and it can be used for guiding continuous improvement of nursing in neonatal surgical department.

Investigation of the factors influencing quality of life in parents with asthma children in community / 现代临床护理

Objective To investigate the life quality of parents of asthma children as well as the influencing factors? Method Totally 125 parents of children with asthma from three different communities in Guangzhou were involved in the survey by the Chinese Paediatric Asthma Caregiver’s Quality of Life Questionnaire (PACQLQ) and demographic questionnaire? Results The total score on PACQLQ was(4?34±1?03): the scores on dimentions of limited motion and affection were(4?29±1?55)and(4?36±0?89),respectively?The major factors influencing the life quality of their parents included the relationship with them and their disease course? Conclusions The life quality of their parents reaches the lowest level at the beginning of confirmed diagnosis of asthma? The health education should be performed right after confirmed diagnosis? During health education,their mental stress is worth our great attention?

Paired basic amino acid cleaving enzyme 4 and tumor / 国际肿瘤学杂志

Paired basic amino acid cleaving enzyme 4 (PACE4),a subtilisin-like endoprotease,which is thought to play a significant role in tumor occurrence and development.Its over or low expression may lead to enhanced proliferation and invasion of tumor cells,and even increase the malignant degree.The specific regulation according to the roles of PACF4 expression in different tumors may be helpful for tumor treatment and prognosis improvement.

Effect of individualized health education to asthmatic children on the quality of life of their parents / 现代临床护理

Objective To explore the influence of individualized health education to asthma children on the quality of life of their parents.Methods One hundred and one parents of asthmatic children from 3 different communities were randomly assigned to the intervention group(n=52)and the control group(n=49).Follow-ups were performed by phone calls and the Children Asthma Health Education Brochures were distributed among them in the two groups,meanwhile the individualized health education was given to the intervention group.The paediatric asthma caregiver's quality of life questionnaire(PACQLQ)was used to assess the quality of life of two groups.Results One month after intervention,the scores of the intervention group on PACQLQ total score as well as its items of activity limitation and emotional function were significantly higher than the control group(P<0.001 for all).Conclusion The individualized health education to the asthmatic children can improve the quality of life of their parents.

Investigation on the effect of performance training of adverse events / 中华医学教育探索杂志

Objective To investigate the application and effect on the performance training of adverse events, and to improve the quality of nursing teaching. Methods Totally 120 new graduate nurses in our hospital were randomly divided into study group(n=60)and control group(n=60). Study group used performance training while control group used traditional training mode and evaluation was made from the adverse event knowledge score , number of adverse events and satisfaction degree of stu-dents. Results Adverse event knowledge score of study group (9.57±0.563) was obviously higher than that of control group(84.90±3.245). Satisfaction degree of students in study group(9.57±0.563) was higher than that of control group(7.35 ±0.917), with statistical differences(P<0.01). Number of ad-verse events(n=3) was obviously lower in study group than in control group(n=15), with statistical differences(P<0.01). Conclusion In adverse events teaching, performance training can be helpful in arousing students' interests in learning. Experiences of self-examination and self-correction also make the students have a deep impression on knowledge, which improves the quality of adverse events teaching and reduces the incidence of adverse events in the new graduates.

Copy number variations of DNA and neoplasms / 国际肿瘤学杂志

The DNA copy-number variant (CNV) is a kind of segments of DNA ranging from 1 kb to 3 Mb that is present in a variable number of copies.CNVs widely distribute across the human genome,and dramatically increases genetic diversity.In recent years,researches have found that most CNVs are closely related to complex diseases.If a cancer gene is directly encompassed or overlapped by a CNV,it may lead to activation of oncogenes or inactivation of tumor suppressor genes,and finally results in tumorigenesis.CNVs can affect gene expression,phenotype differences and phenotypic adaptations by changing gene dosages and gene activities,and then sequentially lead to tumor or any other genetic dieases.Investigating CNVs is apparently helpful for studing chromosome recombination,genomic evolution,gene expression and the pathogenesis of multiple complex diseases especially tumor.

Expression of vitamin D-binding protein in synovial tissues of rheumatoid arthritis / 中华风湿病学杂志

Objective To screen the proteins with decreased expression in the synovial tissues of rheumatoid arthritis (RA) patients by comparing their expression profiles with that of osteoarthritis (OA) and ankylosing spondylitis (AS) patients by a proteomic approach,and to explore the association of reduced expression with disease susceptibility by a single nucleotide polymorphism (SNP) analysis.Methods Proteins extracted from the synovial membranes (n=10 for each disease) were separated by 2-D electrophoresis.The proteins with significantly decreased expression in the RA samples were subjected to MALDI-TOF-MS.The results were verified using Western blotting.Tag SNPs located in the targeted gene were assessed using the Taqman assay in a cohort of 267 Chinese patients with RA and 160 healthy controls.The genotyping results were confirmed in a large cohort of 389 patients with RA and 371 healthy controls.SPSS 11.5 software package was used for one way ANOVA and Fisher's exact test.Results The expression of vitamin D-binding protein (VDBP) in the synovial membranes from patients with RA was significantly decreased when examined by proteomic approach.This result was confirmed by Western blotting analysis.The rs2282679 was significantly associated with RA (P=0.026 794).This result was confirmed in a large cohort of RA( OR=0.678 639,95％CI 0.54l 113-0.851 118,P=0.000 776).Conclusion Compared with samples from patients with OA and AS,RA patients' synovial tissues have low VDBP expression when examined by the proteomic method.The tag SNP rs2282679 located in VDBP is significantly associated with RA.The decreased expression and the genetic effect of VDBP in RA suggest that a novel pathogenic pathway,in which vitamin D contributes,may be involved in the arthritis process of RA.

Correlation of peptidylarginine deiminase 4 gene polymorphism and colorectal cancer / 国际肿瘤学杂志

Objective We aim to investigate the association between peptidylarginine deiminase type 4 (PADl4) and colorectal cancer by genotyping threetag single nucleotide polymorphisms (tagSNP) genetic maker, to explore the role of polymorphism of PAD14 gene in development of colorectal cancer. Methods A case-control study was conducted using TaqMan-PCR methods to analyze the genotypes of three TagSNPs such as rs1886302, rs2477131 and rs1635561 for 515 patients with colorectal cancer and 549 health controls. Results There is significantly different in allelic frequencies and genotype frequencies of rs1886302 between cases and controls (P=0.039 and 0.040,respectively). The OR value of A allele and AA genotype is 0.796 and 0.731,respectively,and A allele may reduce the probability of incidence of colorectal cancer. No association Was found among SNPs of rs2477131 and rs1635561 within intronl and intron 15 in terms of coloretal cancer. Conclusion Rs1886302 on PADI4 might be a susceptible SNP to coloretal cancer.

Association between polymorphisms of CYP3A5,MDR1,COX-2 and chemotherapy outcomes of advanced NSCLC / 国际肿瘤学杂志

Objective To investigate whether genotypes of CYP3A5,MDR1 and cyclooxygenase-2 are associated with the sensitivity of vinorelbine-platinum to NSCLC.Methods The genotypes of CYP3A5(*3),MDR1 (2677G>T at exon 21 and 3435C>T at exon 26 and their haplotypes),cyclooxygenase-2 (-1 195G>A) were determined by RFLP-PCR and chemotherapy responses were analyzed in 69 non-small-celllung cancer (NSCLC) Chinese Han patients.They received a combination chemotherapy of vinorelbine-cispla-tin.Chi-square test was used to investigate the potential association of genotype with chemotherapy response.OR and 95% C1 were calculated.Results The 3435 CC genotype was associated with a significantly betterchemotherapy response compared with the combined 3435 CT and 1Tr genotypes(P=0.033).The 2677 GG genotype was also associated with a significantly better chemotherapy response compared with the combined 2677 GT and IT genotype(P=0.012).Moreover.patients with the 2677 G-3435 C haplotype seemed to have a better response to chemotherapy compared with those with the other haplotypes(P=0.063).CYP3A5*3 was not likely to correlate with sensitivity of vinorelbine-platinum to NSCLC.Cyclooxygenase-2-1195G>A was likely to have better response to vinorelbine but not statistically significant(P=0.067).Conclusion Polymor-Dhisms of MDR1 3435 C>T and MDR1 2677 G>A/T can be used for predicting treatment response to vinorel-bine-cisplatin chemotherapy in NSCLC patients.

Protein-chip for autoantibodies profile detection / 生物工程学报

We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solution, spotting concentration of the 12 antigens and the dilution of serum. We prepared a protein chip that could detect 12 autoantibodies simultaneously using the optimized antigen concentration. We established a new method to determine the cutoff of each autoantibodies by evaluation of 678 positive and 120 negative serum of clinical sample. We also evaluated the sensitivity and specificity of our new detection system. The optimal spotting solution was 0.1% TBST, the dilution of serum was 1:4 and the best spotting concentration of the 12 antigens were ANA 520 microg/mL, Ro-60/SSa 465 microg/mL, La/SSb 530 microg/mL, Jo-1 530 microg/mL, Scl-70 525 microg/mL, Sm 520 microg/mL, Ro-52/SSa 615 microg/mL, RF 340 microg/mL, CCP 465 microg/mL, ulRNP 410 microg/mL, CENP-B 490 microg/mL and dsDNA 580 microg/mL respectively. It had higher coincidence rate compared to current clinical used methods. We have developed a 12 antigens protein chip for the detection of autoantibodies based on the NBT/BCIP color reaction system. This system was fast, convenient, efficient, and cost-effective.

Preparation conditions optimization and preliminary application of the microarray for detecting autoantibodies / 中华风湿病学杂志

Objective To optimize several key factors in preparing protein microarray,and establish microarrays to detect autoantibodies.Methods Four factors,including incubation time of the microarray after spotting,the blocking reagent,the length of blocking time,the length of time conjugating with the second-an- tibody,were selected to carry out orthogonal optimization,then we determined the optimal spotting concentra- tion of antigen and the best dilution of serum with the optimized conditions obtained from the above.On the basis of the optimized conditions,we prepared microarray to detect six autoantibodies simultaneously,which were compared with IIF and ELISA.Results The best conditions we determined from the experiment were: incubating the spot microarray for 120 min,PBST containing 1% BSA and 2.5% saccharose as the blocking reagent and blocking for 30 min,reacting with the second-antibody for 30 min,the appropriate spotting con- centrations of six antigens differed from 100?g/ml to 300?g/ml and the best serum dilution was 1:2.Com- pared with IIF for ANA detection,the sensitivity of the microarray was 88.6%,the specificity was 83.3%,and compared with ELISA in detecting the other five autoantibodies,the sensitivity of the microarray was between 80.0% and 88.2%,and the specificity was between 90.2% and 98.6%.Conclusion The optimization condi- tions we obtained are suitable for preparing protein microarray,and the detection sensitivity and specificity of autoantibodies by microarray are consistent with the conventional methods in general.The microarray for de- tecting autoantibodies is applicable in the clinical diagnosis of autoimmune disease.