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Objective:Serum soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) is a useful biomarker of bacterial infection. However, the diagnostic value of sTREM-1 of alveolar fluid in pulmonary infection is still unclear. This article aimed to explore the value of sTREM-1 of alveolar fluid in the early diagnosis of ventilator-associated pneumonia (VAP) by systematic review of relevant literatures.Methods:CNKI, Wanfang, VIP, PubMed/Medline and Embase databases were retrieved. Articles on diagnosis of VAP by sTREM-1 before June 30, 2019 were collected. QUADAS-2 scale provided by Cochrane Collaboration Network was used to evaluate the quality of diagnostic experiments. RevMan 5.3 and Stata 13.0 software were used to complete Meta-analysis. The levels of sTREM-1 between VAP and non-VAP patients were analyzed by Meta-analysis, and then diagnostic test Meta-analysis was conducted. Heterogeneity, sensitivity and publication bias were analyzed.Results:A total of 24 articles were enrolled. QUADAS-2 scale indicated that the selected literature had low bias and high clinical adaptability. ① In Meta-analysis of sTREM-1 level in alveolar fluid, 20 articles were selected and found to have high heterogeneity ( I2 = 94.4%, P = 0.000). The random effects models were used for Meta-analysis. It was indicated that the sTREM-1 level in alveolar fluid of VAP group was significantly higher than that of non-VAP group with significant difference [standardized mean difference ( SMD) was 1.47, 95% confidence interval (95% CI) was 1.00-1.95, Z = 6.14, P = 0.000]. By subgroup analysis and Meta-regression analysis, no source of heterogeneity was found. Sensitivity analysis suggested that the results of this Meta-analysis were robust and credible, and Begg funnel plot analysis showed that there was no significant publication bias ( Z = 1.46, P = 0.143). ② A total of 18 articles were included in the Meta-analysis of diagnostic experiments. Deek funnel plot showed publication bias ( P = 0.012). The combined sensitivity was 0.87 (95% CI was 0.81-0.91), specificity was 0.80 (95% CI was 0.73-0.86), and diagnostic odds ratio ( DOR) was 26 (95% CI was 13-50). Subgroup analysis of three different sources of alveolar fluid (bronchoalveolar lavage fluid, endotracheal aspiration fluid and exhaled ventilator condensate) showed that STREM-1 had a certain value in early diagnosis of VAP. The I2 of combined DOR was 35.4%, and I2 of sensitivity was 79.46%, I2 of specificity was 77.61%, suggesting heterogeneity in the selected literature. Subgroup analysis found that nationality, subject design, sample source, sample size and diagnostic "gold criteria" were related to heterogeneity, but not age. The area under synthetic receiver operating characteristic (SROC) curve (AUC) was 0.90 (95% CI was 0.87-0.92). Conclusions:The detection of sTREM-1 level in alveolar fluid can be used for the early diagnosis of VAP with high sensitivity and specificity. If combined with other biomarkers, it may have more diagnostic value.
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Objective:To investigate the effect of upregulated and downregulated PSMA7 on the cell cycle and Cyclin D1,CDK4,P16,Rb of RB pathway in A549 cells.Methods:Transfected upregulated pcDNA3.1-PSMA7 vecter and downregulated pGPU6/Hygro-PSMA7-265 vecter into A549 cells,and then tested the effect of PSMA7 on the cell cycle of A549 cells by flow cytometry,and detected the protein level of Cyclin D1,CDK4,P16,Rb by Western blot.Results:Compared with the control group,the cell cycle of the A549 cells did not change significantly,and the expression of Cyclin D1,CDK4 decreased but P16,Rb increased when PSMA7 was upregulated.Compared with the control group,the proportion of phase G0/G1,G2/M of the A549 cells decreased and phase S increased,and the expression of Cyclin D1,CDK4 increased but P16,Rb decreased when PSMA7 was downregulated.There was statistical significance for those results.Conclusion:PSMA7 could affect the expression of Cyclin D1,CDK4,P16,Rb protein level of RB pathway in A549 and promoted the A549 cells into phase S when it′s downregulated.
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Objective To investigate the diagnosis and disease assessment of serum pancreatic stone protein(PSP/reg) in ventilator-associated pneumonia(VAP) by a prospective study.Methods The blood sample was collected in patients with mechanical ventilation(MV) from September 2012 to October 2015.Then the concentration of PSP/reg was detected by ELISA method and the time of MV and the outcome of VAP were recorded,while the patients who did not have VAP occurred in the same period regarded as control group.Results Compared with the control group,there was no significant difference in the serum concentration of PSP/reg in VAP group(P>0.05).From continuous monitoring,compared with the start time of MV,there was a significant increase in the concentration of PSP/reg in VAP group(P0.05).Conclusion PSP/reg can be used as one of the indicators for diagnosis of VAP,and may be related to the severity of VAP.
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Objective: To investigate the expression of HSPC238 in cervical intraepithelial neoplasia and cervical cancer.Methods:We collected 76 cases of cervical cancer,105 cases of CIN and 28 cases of normal cervical epithelial.Then we inves-tigated the expression of HSPC238 by using immunohistochemistry and compared the significant differences between them.Results:There was no significant difference in the expression of HSPC238 between the cervical cancer and normal cervical epithelial ( Z=-0.242,P>0.05).However,there was significant difference between the cervical intraepithelial neoplasia and normal cervical epithelial (χ2=19.159,P<0.01) and the expression of HSPC238 was correlated with the grades of CIN.The expression of HSPC238 decreased when the grade of CIN was increasing.( rs=-0.327,P<0.01 ).Conclusion:The low expression of HSPC238 might be correlated with the development of cervical neoplasia.
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Objective:To investigate the effect of HSPC 238 overexpression or low expression on the regulation of the target protein ( HMOX1, MT2A, UBB, RPS27a ) and the regulation pathways.Methods: To construct the interference vector and overexpression vector of HSPC238 respectively,transfected the HEPG2 cell lines by the liposome method,detect the expression of mRNA and protein of the HSPC 238 and the target proteins by the RT-PCR and Western blot , further to transfer the overexpression plasmids of the target proteins into the HEPG 2 cell lines which had been transfected with interference vector and overexpression vector of HSPC238,the experimental group cell lines were treated with proteasome inhibitor MG 132,to purify the target proteins with nickel column,do immunoblotting with HSPC238 to detect the accumulation situation of the target proteins.Results: The HMOX1,MT2A, RPS27a were downregulated obviously when the HSPC 238 was interfered exogenous;and the MT2A,UBB,RPS27a were up-regulated after the HSPC238 overexpressed.The overexpression plasmid of target proteins were transfected into the HEPG 2 cell lines which have been transfected with interference vector and overexpression vector of HSPC 238 ,compared with the control group ,the target protein band in the experimental group was significantly increased after treatment with the proteasome inhibitor MG 132.Conclusion:The HSPC238 overexpression can upregulate the expression of MT 2A,UBB and RPS27a,and interfering HSPC238 can downregulate the expression of HMOX1,MT2A and RPS27a;the HSPC238 target protein may play a regulatory role through the UPP pathway;the HSPC238 may play a regulatory role on the target proteins through the UPP pathway.
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Objective To investigate the diagnosis and prognosis evaluation value of neutrophil elastase (NE) in ventilator-associated pneumonia (VAP).Methods A retrospective analysis was conducted.The data of patients undergoing mechanical ventilation admitted to Department of Central Intensive Care Unit (ICU) of Boai Hospital of Zhongshan City Affiliated to Southern Medical University from September 2012 to October 2015 were enrolled.The patients were divided into two groups according to whether they suffered from VAP or not.The content of NE in serum and bronchoalveolar lavage fluid (BALF) at the time of mechanical ventilation start,VAP diagnosis (the worst value from 48 hours after mechanical ventilation start to weaning in non-VAP patients),and at the time before mechanical ventilation weaning,as well as inflammation parameters,clinical pulmonary infection score (CPIS),duration of mechanical ventilation and prognosis were recorded.Receiver operating characteristic curve (ROC) was used to analyze the predictive value of NE on VAP diagnosis and prognosis.Results Finally 38 patients were enrolled in the VAP group,and 40 in non-VAP group,and baseline data was similar between the two groups.There was no significant difference in the content of NE in serum and BALF between VAP group and non-VAP group [serum NE (μg/L):67.04 (63.00,75.75) vs.69.00 (63.75,75.00),BALF NE (μg/L):96.26 (85.26,176.01) vs.95.26 (86.76,107.11),both P ＞ 0.05].From continuous monitoring,no significant change in the content of NE in serum and BALF during mechanical ventilation was found in the non-VAP group,but the content of NE in serum and BALF at the time of VAP diagnosis in VAP group was significantly higher than that at mechanical ventilation start [μg/L:157.00 (153.04,165.75) vs.67.04 (63.00,75.75),178.04 (153.00,188.25) vs.96.26 (85.26,176.01),both P ＜ 0.05],and NE content in serum and BALF was significantly decreased at the time after VAP clinical recovery and before mechanical ventilation weaning [μg/L:75.67 (64.51,110.55) vs.157.00 (153.04,165.79),95.50 (66.56,183.02) vs.178.04 (153.00,188.25),both P ＜ 0.05].The NE in the start time of VAP in VAP group was divided into four groups according to quartile,it was found that with the increase of NE content in serum and BALF,the CPIS was increased,the duration of mechanical ventilation was prolonged,and the prognosis was poor (all P ＜ 0.01).Compared with non-VAP group,white blood cell count (WBC),neutrocyte proportion,C-reactive protein (CRP),and procalcitonin (PCT) in VAP group were significantly increased (all P ＜ 0.01).NE in BALF was significantly positively correlated with WBC,neutrocyte proportion,CRP and PCT (r value was 0.507,0.432,0.779,and 0.519,respectively,all P =0.000),among which the highest correlation was CRP.NE in BALF used for VAP diagnosis has good accuracy,with sensitivity of 87.4％,and specificity of 90.6％,and sensitivity and specificity of NE in serum for VAP diagnosis was 78.6％ and 79.2％ respectively.Conclusion NE can be used as one of the indicators for VAP diagnosis,and it is related to the prognosis of VAP.
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Objective To investigate the effect of up‐regulation and down‐regulation of HSPC238 on the RB gene mRNA and pRb protein .Methods PcDNA3 .1‐HSPC238 vector and PLL3 .7‐HSPC238 vector was transiently transfected into HepG2 cells re‐spectively .The level of RB mRNA was detected by real‐time PCR and pRb was detected by Western blotting .Results The level of RB mRNA and pRb in up‐regulation group both increased compared with control grops respectively .Conversely ,the level of RB mRNA and pRb in up‐regulation group both decreased compared with control grops respectively .Above results were all statistically significant(P<0 .05) .Conclusion HSPC238 could have a positive effect on the expression of the RB gene .
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Objective:To construct a bait vector for HSPC238, and to screen the target proteins which interact with the HSPC238.Methods:Gene synthesis method was used to synthetic gene HSPC238, then connected with the pGBKT7 vector after digesting by the sfiIA and sfiIB,to obtain the bait plasmid pGBKT7-HSPC238,then transferred into the yeast strains AH109 with the empty plasmid pGBKT7after sequencing,to observe its self-activating effect in the nutrient deficiencies medium,and further to screen the target proteins which interact with HSPC238 from the human fetal liver cDNA library.Results:The bait vector pGBKT7-HSPC238 was successfully constructed,and it had no self-activating effect through the phenotypic screening,after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5(RPL5) may be the one of the target proteins which interacted with HSPC238 from the human fetal liver cDNA library.Conclusion: We successfully constructed the bait plasmid vector PGBKT7-HSPC238,and after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5( RPL5) may be the one of the target proteins which interacted with HSPC238.
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Objective To evaluate the diagnostic value of serum α-L-fucosidase (AFU)in primary hepatic carcinoma(PHC)by using the meta analysis method.Methods The databases of Cochrane Library,PubMed,web of knowledge(Medline,BIOSIS Pre-views),Springer link and Science Direct were retrieved from January 1997 to December 2013.The domestic databases of CBMdisc (Chinese BioMedical Literature on disc),CNKI,Wangfang and VIP(VIP Database for Chinese Technical Periodicals)were retrieved from January 1984 to December 2013.Retrieval languages were limited to Chinese and English.The related literatures on the study of serum AFU diagnostic value for PHC were collected and the included studies meeting the inclusion criteria were performed the quality assessment.The meta-Disc 1 .4 software was adopted to conduct the analysis for comprehensively evaluating serum AFU value in the diagnosis of PHC.Results 20 articles were included (15 articles in Chinese and 5 articles in English).A total of 2 114 cases in the patients groups and 5 718 cases in the control groups were analyzed.The heterogeneity test was carried out on the in-cluded 20 articles,suggesting that the heterogeneity of the included studies was caused by the non-threshold effect,so the random effects model was selected for conducting the meta-analysis.And the pooled sensitivity was 0.77,the pooled specificity was 0.87, the pooled positive likelihood ratio was 5.37,the pooled negative likelihood ratio was 0.28,the pooled diagnostic odds ratio was 20. 47 and the SROC area under the curve(AUC)was 0.87.Conclusion Serum AFU has highly sensitivity and specificity for the diag-nosis of PHC and has reference value for its clinical diagnosis.
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Objective:To investigate the association between rs1042713 polymorphisms of ADRB2 gene and the susceptibility of asthma in Chinese Population by meta-analysis.Methods: The Pubmed database,Emabase database,Web of Knowledge database, CNKI database,Wanfang database and Weipu database were searched for all publications about the susceptibility of asthma and the rs1042713 polymorphisms of ADRB2 gene in Chinese Population.The article which met the inclusion criteria were assessed by the STA-TA12.0 software.Results:12 studies were included,with 2 193 asthmatic patients and 2 033 controls.All the included articles were satisfied to the Hardy-Weinberg equilibrium.The results of Meta-analysis was showed that the risk of asthma of the mutations G carriers ( GG +GA) on ADRB2 gene rs1042713 loci in Chinese people compared with the wild-type homozygotes( AA) was not significantly in-creased overall(OR=1.08,95% CI=0.82-1.44).However,subgroup analysis showed that the risk of G carriers of children had a relatively higher incidence(OR=1.69,95% CI 0.99-2.87),while the risk of adult-onset have a relatively lower incidence(OR=0.88,95%CI 0.68-1.15).Conclusion:The ADRB2 gene rs1042713 polymorphisms have a certain correlation with the susceptibility of asthma in Chinese children,the mutant gene G carriers may relatively increase the risk of asthma in childhood.
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Objective:To construct the recombinant eukaryotic expression vector pCDNA3.1-MT2A and to investigate the cellular localization of MT2A protein in 293T and SMCC7721cell lines.Methods: Gene synthesis method was used to synthetic gene MT2A,added a Kozak sequence and His tag sequence at the N-terminus,the amplified target gene was connected to the pcDNA3.1(+) vector which was double digested between the BamH Ⅰ and Not Ⅰ.After transformation to E.coli DH5α, the positive clones were picked for plasmid extraction then Electrophoretic and sequenced.The pCDNA3.1-MT2A plasmids which passed through electrophoretic and sequenced were transfected 293T and SMMC7721 cell lines by liposome method,and then observed their expression and localization in eukaryotic cells by laser confocal microscopy.Results: The recombinant plasmid pCDNA3.1-MT2A was confirmed by restriction analysis and DNA sequencing,the sequence of the target gene MT2A was entirely correct,eukaryotic expression vector was successfully constructed and cell lines which had transfected recombinants could see the expression of green fluorescent protein in the cytoplasm.Conclusion:Successfully constructed fusion gene of pCDNA3.1-MT2A and expressed in eukaryotic cells,we found that the MT2A was mainly localized in the cytoplasm of 293T and SMMC7721 cell lines.The findings can help us to lay the foundation for the functions of MT2A in hepatoma cells.
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Endothelial monocyte-activating polypeptide-Ⅱ (EMAP-Ⅱ ) is a novel proinflammatory cytokine with proinflammatory,proapoptotic and antiangiogenic properties.It is associated with many tumorassociated proteins,such as tumor necrosis factor,vascular endothelial growth factor,hypoxia inducible factor-1α,arginyl-tRNA synthetase,and insulin-like growth factor- Ⅰ.EMAP-Ⅱ has anti-tumor properties and has a good prospect in cancer prevention and treatment.
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0.05). The data proved that PSMA7 was overexpressed in pcDNA3.1(-)/PSMA7-transfected A549 cell line, both in mRNA level and in protein level. Conclusion Eukaryotic expression plasmid pcDNA3.1(-)/PSMA7 has been successfully constructed, and the PSMA7 is stably expressed in A549 cell line. This would pave the way for further study of PSMA7.