Clinical diagnostic value of presepsin in elderly patients of acute left heart failure complicated with pneumonia / 中华急诊医学杂志

Objective:To investigate the clinical diagnostic value of soluble leukocyte differentiation antigen 14 subtype (sCD14-ST, presepsin) in elderly patients with acute left heart failure (AHF) complicated with bacterial pneumonia.Methods:The data of 111 elderly patients with acute left heart failure complicated with bacterial pneumonia or acute left heart failure (the control group) who were admitted into emergency department from August 2017 to August 2018 were retrospectively analyzed. Chemilluminescence immunoassay was performed to detect presepsin in all patients. And meanwhile, fever or not, presepsin, procalcitonin (PCT), C-reaction protein (CRP) and other clinical data were compared between the two groups. Univariate and multivariate logistic regression analysis were adopted to screen the risk factor influencing the diagnosis. The receiver operating characteristic (ROC) curve was used to analyze the clinical value of presepsin on diagnosing acute left heart failure complicated with bacterial pneumonia in elderly patients.Results:Presepsin of the group complicated with bacterial pneumonia was significantly higher than that of the control group [(500.9±283.5) ng/L vs. (167.7±102.3) ng/L, t=-7.902, P=0.000]. The logistic regression analysis, showed that fever, presepsin and procalcitonin were independent risk factors for AHF combined with bacterial pneumonia diagnosis. The area under ROC curve (AUC) of presepsin, PCT and WBC was 0.887 (95% CI: 0.825-0.949, P<0.001), 0.794(95% CI: 0.704-0.885, P<0.001), and 0.566 (95% CI: 0.455-0.678, P=0.231), respectively. The optimal threshold value of presepsin was 227 ng/L, the sensitivity was 82.0%, specificity was 83.6%, the positive likelihood ratio was 5, the negative likelihood ratio was 0.22, the positive predictive value) was 80.4%, and the negative predictive value was 85%. Conclusions:Presepsin has an important diagnostic value for the identification of AHF combined with bacterial pneumonia in elderly patients.

Effect of Shaoyaotang on Expressions of CD14, FADD and Caspase-8 in Colonic Tissues of Rats with Large Intestinal Damp-heat Syndrome of Ulcerative Colitis / 中国实验方剂学杂志

Objective:To observe the effect of Shaoyaotang on the contents of cell adhesion molecule-1 （ICAM-1） and transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁） in serum of large intestine damp-heat syndrome of ulcerative colitis （UC） in rats， and the gene and protein expressions of leukocyte differentiation antigen14 （CD14）， Fas-related death domain protein （FADD） and cysteinyl aspartate specific protease-8 （Caspase-8） in the focal colon tissue. Method:A total of 80 SPF Wistar rats were randomly divided into the blank group （<italic>n</italic>=10） and modeling group （<italic>n</italic>=70）. The large intestine damp-heat syndrome of UC rats was replicated by the combination of disease and syndrome， which was high-fat， high-sugar and spicy diets combined with 2， 4-dinitrobenzene sulfonic acid （DNBS） and ethanol. After successful modeling， the modeled groups were divided into model group， sulfasalazine （SASP）control group， and low， medium and high-dose Shaoyaotang groups by the method of random number table， with14 rats in each group. Low， medium and high doses of Sulfasalazine 0.2 g·kg^-1·d^-1 and Shaoyaotang （6， 12， 24 g·kg^-1·d^-1）were given by gavage. The blank group and the model group were given equal volume of normal saline for 21 days. The contents of serum ICAM-1 and TGF-<italic>β</italic>₁ were detected by enzyme-linked immunosorbent assay （ELISA）， the expressions of CD14， FADD and Caspase-8 mRNA in colon tissues were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）， and the expressions of CD14， FADD and Caspase-8 protein in colon tissues were detected by Western blot. Result:Compared with the blank group， the serum ICAM-1 level in the model group were significantly increased， whereas the content of TGF-<italic>β</italic>₁ were significantly decreased （<italic>P</italic><0.05）. The relative expression levels of CD14， FADD， Caspase-8 mRNA and protein were significantly increased （<italic>P</italic><0.05）. Compared with the model group， the content of ICAM-1 in the serum of the rats in the medium， high-dose Shaoyaotang groups and the SASP group were significantly decreased， while the content of TGF-<italic>β</italic>₁ in the serum of the rats in the low， medium， high-dose Shaoyaotang groups and the SASP group were significantly increased （<italic>P</italic><0.05）. The expression levels of CD14， FADD， Caspase-8 mRNA and protein in each intervention group were significantly decreased （<italic>P</italic><0.05）， especially in the high-dose Shaoyaotang group and the SASP group. Conclusion:Shaoyaotang has a certain intervention effect on UC rats with large intestine damp-heat syndrome， and its mechanism may be related to the inhibition of CD14， FADD and Caspase-8 genes and proteins expression.

Serum Presepsin in the diagnosis and assessment of sepsis in children / 中国小儿急救医学

Objective:To investigate clinical significance of Presepsin(soluble CD14 subtype) in the diagnosis and condition assessment of sepsis in children compared with traditional biomarkers.Methods:For the prospective study, 102 children with sepsis admitted to the PICU of the Children′s Hospital of the Capital Institute of Pediatrics from January 2017 to December 2018 were selected, including 57 cases in the sepsis group, 45 cases in the severe sepsis/septic shock group and 25 cases in the non-infectious systemic inflammatory response syndrome(SIRS group), and 35 children with healthy physical examination during the same period as the control group.The sepsis group was further divided into the survival group( n=86)and the death group( n=16)based on the 28-day mortality.The data collected included serum Presepsin, procalcitonin(PCT), C-reaction protein(CRP) and interleukin(IL)-6 levels on days 1, 3 and 7 of admission, and compared with paediatric critical case scores. Results:(1)The levels of serum Presepsin [12.43(7.21, 15.07) ng/mL], PCT [23.00(5.70, 87.00) ng/mL], CRP [160.0(105.5, 200.0) mg/L], IL-6 [1 000.0(125.0, 1, 000.0) pg/mL] were significantly higher than those in the sepsis, SIRS and control groups( P<0.001). (2) The area under the ROC curve(AUC) values for Presepsin, PCT, and IL-6 subjects on day 1 of admission were 0.856, 0.812, and 0.516, respectively.The sensitivity of Presepsin at a cut-off value of 4.40 ng/mL for the diagnosis of sepsis was 81.1% and the specificity was 72.3%, which would significantly higher diagnostic efficacy of the combination of Presepsin, PCT and IL-6.(3) There was a significant difference between the survival and death groups in Presepsin( P<0.001), and Presepsin was significantly higher in the death group on days 3 and 7 than those in the survival group(both P<0.001); IL-6 was significantly higher in the death group on day 3 than that in the survival group( P=0.04); the differences in PCT and CRP between the death and survival groups at all time points were not statistically significant(both P>0.05 ). (4) The AUCs of inflammatory factors on days 1, 3 and 7 to predict sepsis outcome were 0.597, 0.656 and 0.951 for Presepsin, 0.576, 0.613 and 0.655 for PCT and 0.726, 0.786 and 0.664 for IL-6, respectively.The diagnostic values of Presepsin on day 7 and IL-6 on days 1 and 3 were higher.The combination of Presepsin, PCT and IL-6 significantly improved the prognostic judgment of sepsis.(5) The difference between sepsis-related acute kidney injury(AKI) and non-AKI was not statistically significant when comparing Presepsin on day 1 and 3(all P>0.05). Presepsin levels on day 7 were significantly higher in children with sepsis-associated AKI than in those without AKI( P<0.001). Conclusion:Presepsin is a good biomarker for sepsis diagnosis in children, which is equivalent to PCT in the diagnosis of sepsis, superior to IL-6 and superior to PCT in the prognosis evaluation.Combined testing of Presepsin, PCT and IL-6 may improve the diagnosis of sepsis and the assessment of the condition in children.

LRRC25 plays a key role in all-trans retinoic acid-induced granulocytic differentiation as a novel potential leukocyte differentiation antigen

Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.

Clinical diagnostic value of presepsin in patients with rheumatoid arthritis complicated with bacterial infection / 中华风湿病学杂志

Objective To investigate the clinical diagnostic value of plasma soluble leukocyte differentiation antigen 14 (presepsin) in patients with rheumatoid arthritis (RA) complicated with pulmonary bacterial infection.Methods A total of 133 patients with RA and 60 healthy controls were enrolled in this study.Fifty-eight RA patients were infected with lung bacterial infection and 75 patients were non-infected with RA.Among them,RA activity was performed in 43 patients and RA was stable in 32 patients.Chemilluminescence immunoassay was used to detect presepsin (P-SEP) in all subjects,and its correlation with inflammatory markers such as white blood cells,blood sedimentation rate and C-reactive protein was analyzed.Results The P-SEP of RA (561 ±142) pg/ml combined with pulmonary bacterial infection group was significantly higher than that of active RA group (378±100) pg/ml (t=8.12,P＜0.01),higher than that of stable RA group (197±68) pg/ml (t=8.51,P＜0.01) and healthy control group (113±9) pg/ml (t=13.75,P＜0.01).There was a positive correlation between P-SEP and leukocyte count in patients with RA complicated with pulmonary bacterial infection (r=0.627,P＜0.01).The degree of the disease activity was correlated with CRP (r=0.63,P＜O.O1),regardless of the P-SEP level (r=0.47,P=0.521).The optimal threshold value of P-SEP in diagnosing RA patients with bacterial infection was 458.9 pg/ml,with a sensitivity of 79.3％ and a specificity of 81.4％.Conclusion P-SEP has an important diagnostic value for the identification of bacterial infection in RA patients,which is not related to RA disease activity.

Role of Mac-1 in osteoclast differentiation / 中国病理生理杂志

AIM:To investigate the function of macrophage differentiation antigen-1 (Mac-1) in receptor acti-vator of nuclear factor-κB ligand ( RANKL )-induced osteoclast differentiation and the mechanisms .METHODS: The spleen cells were isolated from 4-week-old C57BL/6J mice, and cultured with RANKL, macrophage colony-stimulating fac-tor and CD11b and CD18 antibodies for 1 week.The actin bundles were stained with rhodamine-labeled phalloidin, and nu-clei were stained with DAPI.CD11b and CD18 antibodies, lentivirus with interfering vector plasmid of target gene Itgam (encoding CD11b) and empty virus (control virus) were used to treat osteoclasts for 1 week, and then immunofluorescence staining was performed.CD11b antibody, lentivirus and control virus were used to treat osteoclasts for 1 week, and total protein was taken for Western blot .RESULTS:Lower multinuclear positive rates in the groups treated with CD 11b anti-body were observed than that in the groups treated with CD 18 antibody and control group .Lower immunofluorescence inten-sity of Syk, CD11b and NFATc1 was found in CD11b antibody group than that in control group .Lower Syk, CD11b and NFATc1 immunofluorescence intensity was also observed in lentivirus group than that in control virus group .The results of Western blot analysis showed that the protein levels of CD 11b, Syk, NFATc1, c-Fos and p-ERK/ERK in CD11b antibody group were decreased as compared control group .Compared with control virus group , the protein levels of CD11b, Syk, NFATc1, c-Fos and p-ERK/ERK in lentivirus group were also decreased .CONCLUSION:CD11b subunit of Mac-1 pro-motes osteoclast differentiation by up-regulating c-Fos, ERK activity and NFATc1.

Effects of DNA methylation of thymocyte differentiation antigen-1 in beryllium sulfate induced human fetal lung fibroblast fibrosis / 中国职业医学

OBJECTIVE: The change of DNA methylation of thymocyte differentiation antigen-1( Thy-1) was observed in beryllium sulfate( Be SO4) stimulated human fetal lung fibroblast( MRC-5 cell) to explore the effects of Thy-1 in Be SO4 induced lung fibrosis. METHODS: MRC-5 cell culture in vitro model was used. The final concentrations of Be SO4were1. 0,10. 0 and 100. 0 μmol / L( low-,medium- and high-dose groups). The control was untreated. Other 2 intervention groups were the 5-azacytidine( AZC) intervention group( 10. 0 μmol / L of AZC and 10. 0 μmol / L Be SO4) and the trichostatin A( TSA) intervention group( 0. 5 μmol / L of TSA and 10. 0 μmol / L Be SO4). The cells were collected 24,48 and 72 hours after exposure. Real-time quantitative polymerase chain reaction( PCR) was used to determine the relative expression of collagen typeⅠ( Col Ⅰ),collagen type Ⅲ( Col Ⅲ),α-smooth muscle actin( α-SMA) and Thy-1 mRNA.The nested landed methylation specific PCR was used to detect the Thy-1 DNA methylation level. RESULTS: At 24 hours,the relative expression level of Col Ⅲ mRNA in MRC-5 cells showed an increasing trend with increasing dose( P < 0. 05);at 48 and 72 hours,the relative expression levels of Col Ⅰ,Col Ⅲ and α-SMA mRNA in MRC-5 cells increased with the increasing dose( P < 0. 05). All these 3 indicators in MRC-5 cells of 3 dose groups increased with the increase of expose time( P < 0. 05). The relative expression level of Thy-1 mRNA in MRC-5 cells of all 3 dose groups were lower than that in control( P < 0. 05). The relative expression level of Thy-1 mRNA of the high-dose group was lower than that of the lowdose group( P < 0. 05). The Thy-1 DNA methylation levels in the medium- and high-dose groups were both higher than that of the control( P < 0. 05). The Thy-1 DNA methylation levels of the 3 dose groups increased with the increasing dose( P < 0. 05). The Thy-1 DNA methylation levels of MRC-5 cells in the 2 intervention groups were higher than that of the control( P < 0. 05),but there was no significant difference when compared with the medium-dose group( P > 0. 05).CONCLUSION: Be SO4 stimulation can induce the fibrosis of MRC-5 cells. In this process,the Thy-1 DNA methylation level increases,while the Thy-1 mRNA expression level decrease. Thy-1 DNA methylation might be one of the important mechanisms of lung fibrosis induced by Be SO4.

The prognosis and relationship between soluble differentiation antigen of cluster designation 14 and the injury severity score of patients with multiple injuries / 中华急诊医学杂志

ObjectiveTo explore the prognosis and relationship between soluble differentiation antigen of cluster designation 14 (sCD14) and the injury severity score (ISS) of casualties with multiple injuries.Methods A total of 86 casualties with multiple injuries were enrolled from October 2009 to March 2010,and the severity of trauma of casualties were assessed in ISS score within the first 24 hours after accident,and the patients were divided into survival and non-survival groups as per the outcomes in 28 days after accident. Another 20 healthy subjects served as control group. In multiple injuries group,sCD14 concentrations were detected and APACHE Ⅱ scores were calculated on the 1st,3rd,5th and 7th days after accident.The related coefficient between sCD14 and ISS was calculated and then the values of sCD14 in predicting prognosis were analyzed by ROC curve. ResultsCompared with control group, sCD14 concentrations were obviously higher in casualties of multiple injuries group at all observation intervals ( P ＜0.01). Compared with survival group, casualties of non-survival group had more higher sCD14 concentrations and APACHE Ⅱ scores ( P ＜ 0.05 ).The sCD14 concentration was correlated with ISS and the related coefficient was 0.469 ( P ＜ 0.01 ). ROC curve analysis suggested sCD14 had values in predicting prognosis of casualties with multiple injuries and area under ROC curve was 0.820.Conclusions The concentration of sCD14 is correlated with ISS and can be used for predicting the prognosis of casualties with multiple injuries.

Establishment of Flow-FISH method for simultaneous detection of telomere length and cell differentiation antigen / 中华微生物学和免疫学杂志

Objective To establish the Flow-FISH method for simultaneous detection of telornere length and cell differentiation antigen. Methods HL60, Raji, Molt4 cells were cultivated. Each step and the conditions of the Flow-FISH procedure were optimized, standardized and validated, then 14 acute leukemia patients were observed for the changes of telomere length combined with differentiation antigen after complete remission by the method. Results Cells were stained with Alexa Fluor(R) 647-labeled antibody. Anti-gen-anfibedy complexes were covalenfly cross-linked onto the cell membrane before telomere staining. Cells were hybridized with telomere-specific fluorescein isothiocyanate (FITC)-conjugated peptide nucleic acid (PNA) probes followed by being counterstained with propidium iodide(PI). Multicolor Flow-FISH was performed to analyze telomere length and differentiation antigen simultaneously. The patients showed longer telomere and lower antigen expression after complete remission. Conclusion The Flow-FISH method for simultaneous detection of telomere length and differentiation antigen was successfully established which might prove to be a promising means for leukemia research especially in those without special molecular markers.

Effect of specific short hairpin RNA of nucleostemin on differentiation antigens and biological properties of HL-60 cells / 第三军医大学学报

Objective To study the role of the specific short hairpin RNA(NS-shRNA) of nucleostemin in inhibiting the expression of nucleostemin(NS) gene and evaluate the effect of NS-shRNA on differentiation antigen and biological properties of HL-60 cells,so as to explore the relationship between the biological functions of NS and acute leukemia,the potential perspective of NS gene therapy with RNA interference(RNAi).Methods HL60 cells were directly transfected NS-shRNA by using special transfection reagent.After 48 h,the inhibition rate of NS-shRNA to NS gene was detected by RTPCR,the proliferation ability of HL-60 cells was detected by MTT,the differentiation antigens were assayed by flow cytometry(FCM),the morphologic peculiarities such as cell shape,growth condition were observed,and the volume and granularity of HL-60 cells were analyzed by blood cell analyzer.Results The expression of NS gene were significantly inhibited by both two NS-shRNA,the inhibition rate were 37.82% and 71.88%,respectively.The significant inhibition effect of NS-shRNA to the proliferation of HL-60 cells existed in a time-dependent and concentration-dependent manner,and the best time was 48 h,the best concentration was 10 nmol/L.The change of differentiation antigen included increase of CD11b,CD33,CD14,CD64,HLA-DR and decrease of CD38,indicating continuous maturation of HL-60 cells to granulocytes and redifferentiation trend to monocytes.The aggregation of cells declined and the cell fragments increased.Myeloperoxidase(MPO) and ?-naphthol acetate esterase(?-NAE) activity increased after NS-shRNA-2 transfection.Furthermore,some HL-60 cells changed from round to fusiform shape even to pseudopod.The cells of small volume and karyorrhexis increased. Conclusion Through blocking the expression of NS gene,NS-shRNA can inhibit the cell proliferation and induce the differentiation of HL-60 cells,weaken malignant degree of HL-60 cells and trigger cell apoptosis.NS-shRNA is of clinical potential in gene therapy for acute leukemia.

Immature thymocyte antigen, JL1, as a possible immunodiagnostic and immunotherapeutic target for leukemia

The identification of tumor-specific antigens has represented a critical milestone in cancer diagnosis and therapy. Clinical research in this area for leukemia has also been driven over the past few decades by the hope that surface antigens with restricted tissue expression would be identified. Disappointingly, only a small number of the leukemic antigens identified to date, meet sufficient criteria to be considered viable immunophenotypic markers. In this paper, we nominate anti-JL1 monoclonal antibody as an immunodiagnostic and immunotherapeutic candidate for leukemia. The JL1 molecule appears to be a novel cell surface antigen, which is strictly confined to a subpopulation of limited stages during the hematopoietic differentiation process. Despite the restricted distribution of the JL1 antigen in normal tissues and cells, anti-JL1 monoclonal antibody specifically recognizes various types of leukemia, irrespective of immunophenotypes. On the basis of these findings, we propose JL1 antigen as a tumor-specific marker, which shows promise as a candidate molecule for diagnosis and immunotherapy in leukemia, and one that spares normal bone marrow stem cells.

Electroporation transfer of human leukocyte differentiation antigenes CD_4, CD_(19) genes into mammalian cells / 中国免疫学杂志

Human leukocyte differentiation antigens CD4 and CD19 gene were transfected into Cos cell line by electroporation method. Indirect immunoflucence and APAAP immunocytochemical staining shows that transient expression and highefficiency transformation of CD4, CD19 genes was obtained. It can provide a new way for reserch on molecular strcture and function of IL-2 receptor gene, identification and screen of Anti-IL-2 receptor McABs.