Effect of Tanxiang Qingyan Tablets on MyD88/NF-κB/ICAM-1 signaling pathway in rats with chronic bronchitis / 中国药理学通报

Aim To explore the effect of Tanxiang Qingyan Tablets on rat model with chronic bronchitisand the effect of MyD88/NF-κB/ICAM-1 signaling pathway expression in bronchopulmonary tissues of rats.Methods A rat model of chronic bronchitis was established by smoking method combined with lipopolysaccharide(LPS,2g·L-1)tracheal injection.The rats were randomly divided into normal group,sham operation group,model group,and positive drug(Guilong Ruikening tablets)1 g·kg-1)group,Tanxiang Qingyan Tablet high,medium and low dose(1.44,0.72,0.36 g·kg-1)group,with intragastric interventionin continuous 15 days.The pathological changes of bronchopulmonary tissues were observed by HE staining,and the infiltration of bronchial inflammatory cells was counted; ELISA method was used to detect the contents of tumor necrosis factor(TNF-α)and interleukin 10(IL-10)in peripheral serum; Western blot and immunohistochemical methodswere employed to detectmyeloid cell differentiation protein 88(MyD88),nuclear transcription factor-κB(NF-κB)and anti-intercellular adhesion molecule 1(ICAM-1)protein expression in bronchopulmonary tissues.Results Compared with normal group and sham operation group,the bronchial mucosal epithelial cells of model group were severely damaged,the alveolar septum was widened,the bronchial inflammatory infiltrationsignificantly increased,the serum TNF-α levels significantly increased,IL-10 levels decreased, and MyD88,NF-κB and ICAM-1 protein expression levels increased significantly(P<0.05,0.01)in bronchopulmonary tissues; compared with model group,the pathological damage and inflammatory changes of the bronchopulmonary tissues of rats in Tanxiang Qingyan Tablet group were reduced,and the serum TNF-α content was significantly reduced,IL-10 content did not change significantly,and MyD88,NF-κB and ICAM-1 protein expression levels in bronchopulmonary tissues were significantly down-regulated(P<0.05,0.01).Conclusions Tanxiang Qingyan Tablets can effectively improve bronchopulmonary tissue inflammatory infiltration,which may be related to reducing the release of inflammatory mediators such as TNF-α and regulating the expression of MyD88/NF-κB/ICAM-1 signaling pathway.

Role and regulatory mechanism of triggering receptor expressed on myeloid cells 2 in mice lung ischemia/reperfusion injury / 中华危重病急救医学

Objective:To investigate the role and regulatory mechanism of triggering receptor expressed on myeloid cell 2 (TREM2) in mice lung ischemia/reperfusion injury (LIRI).Methods:Thirty-six healthy male C57BL/6 mice were divided into six groups according to the random number method ( n = 6): normal control group, and LIRI 2, 6, 12, 24, 48 hours group. Mice LIRI models were established by clamping the left hilum. The wet/dry weight ratio (W/D) of left lung tissue was measured. Lung injury was observed and evaluated by hematoxylin-eosin (HE) staining and electron microscopy. The levels of interleukins (IL-1β, IL-18) in lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of TREM2 and caspase-1 were determined by polymerase chain reaction (PCR). The protein expressions of TREM2, caspase-1, Gasdermin-D (GSDMD) were determined by Western blotting. Results:At 2 hours after LIRI, lung injury began to appear, the lung ultrastructure changed, and the lung injury score increased; at 6 hours, the degree of lung injury was the most serious; after 12 hours, the lung injury gradually reduced and the lung injury score gradually decreased. Compared with the normal control group, lung W/D ratio and lung injury score of LIRI 2, 6, 12, 24, 48 hours groups were significantly higher, the differences were statistically significant (lung W/D ratio: 7.06±0.52, 8.34±0.17, 6.42±0.35, 5.34±0.25, 5.59±0.45 vs. 4.69±0.23; lung injury score: 5.50±0.54, 9.75±0.89, 5.88±0.84, 3.63±0.74, 4.13±0.64 vs. 1.13±0.35, all P < 0.05). Compared with the normal control group, the levels of IL-1β and IL-18 in lung tissue were significantly increased at 2 hours after LIRI, reached a peak at 6 hours [IL-1β (ng/L): 502.76±12.25 vs. 56.50±8.07, IL-18 (ng/L): 414.02±10.75 vs. 81.63±5.29, both P < 0.05], then decreased gradually, and were still significantly higher than the normal control group at 48 hours. The PCR and Western blotting showed that the expression of TREM2 was significantly lower than that in the normal control group at 2 hours after LIRI, and reached a valley at 6 hours [TREM2 mRNA (2 -ΔΔCt): 0.47±0.05 vs. 1.02±0.05, TREM2/GAPDH: 0.23±0.13 vs. 0.48±0.17, both P < 0.05], then gradually increased, and reached the peak at 24 hours [TREM2 mRNA (2 -ΔΔCt): 3.98±0.15 vs. 1.02±0.05, TREM2/GAPDH: 0.71±0.17 vs. 0.48±0.17, both P < 0.05]. The trend of expression of caspase-1 and GSDMD were opposite to that of TREM2, which increased at first and then decreased, and reached a peak at 6 hours after reperfusion [caspase-1 mRNA (2 -ΔΔCt): 2.20±0.13 vs. 1.01±0.02, caspase-1/GAPDH: 0.64±0.02 vs. 0.20±0.06, GSDMD/GAPDH: 1.23±0.01 vs. 0.87±0.02, all P < 0.05]. Conclusions:TREM2 might be involved in LIRI in mice. The mechanism may be related to the effect of TREM2 on caspase-1-mediated pyroptosis.

Effect of Bushen Huatan Prescription on Serum LPS and TLR4/MyD88/NF-κB Signaling Pathway in Rats with Ovariectomy-induced Osteoporosis / 中国实验方剂学杂志

Objective:To explore the effect of Bushen Huatan prescription on serum lipopolysaccharide （LPS） and Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88）/nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） signaling pathway in rats with ovariectomy-induced osteoporosis. Method:Sixty SPF 6-month-old female rats were randomly divided into sham operation group， model group， estradiol valerate group and Bushen Huatan prescription low， medium and high dose groups.One week after modeling by bilateral ovariectomy， 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg^-1 by gavage， and Bushen Huatan prescription low， middle and high dose groups were given 4.7， 9.4 and 18.8 g·kg^-1 by gavage， sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention， the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay （ELISA）， while protein expressions of TLR4， MyD88 and phosphorylated （p）-NF-<italic>κ</italic>B p65 in bone tissue were detected by Western blot， and the mRNA expressions of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction（PCR）. Result:Compared with sham operation group， the serum LPS level as well as protein expression of TLR4， MyD88， p-NF-<italic>κ</italic>B p65 and mRNA expression of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 significantly increased in model group（<italic>P</italic><0.05）.Compared with the model group， serum LPS level， protein expression of TLR4， MyD88， and p-NF-<italic>κ</italic>B p65， mRNA levels of TLR4， MyD88， and NF-<italic>κ</italic>B p65 in bone tissues as well as downstream inflammatory factors IL-1<italic>β</italic>， IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group（<italic>P</italic><0.05）. Conclusion:Bushen Huatan prescription can reduce serum LPS content， regulate mRNA and protein expression of TLR4， MyD88， NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in TLR4/MyD88/NF-<italic>κ</italic>B pathway， and down-regulate mRNA levels of IL-1<italic>β</italic> and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis （PMOP）.

Triggering receptor expressed on myeloid cell-2 modulating the polarization of mouse M2 microglia with oxygen-glucose deprivation/re-oxygenation model / 解剖学报

Objective To investigate the mechanism of TREM2 modulating the polarization of M2 microglia treated by oxygen-glucose deprivation/reoxygenation (OGD/R). Methods Mouse N9 microglial cells were cultured in vitro. N9 cells were transfected with lenti virus for TREM-2-overexpression (LV-TREM2), and LV-scramble acted as control group. OGD/R model was established. The OGD/R cells were randomly divided into OGD/R, OGD/R+LV-scramble and OGD/R+LV-TREM2 groups. Real-time PCR was used to detect the expression of TREM2 mRNA in OGD/R N9 cells within 72 hours after re-oxygenation. Immunofluorescence was applied to observe transfection of lentivirus LV-scramble and LV-TREM2 for normal N9 microglia, and Real-time PCR and Western blotting were used to verify the efficiency of lentivirus transfection. The mRNA and protein contents of Ml microglial markers tumor necrosis factor-a (TNF-α), interleukin-lβ (IL-lβ) and inducible nitric oxide synthase (iNOS), M2 microglial markers arginase-1 (Arg-1) and interleukin-10 (IL-10) were detected by Real-time PCR and ELISA. The expressions of phosphorylated-phosphatidylinositol 3-kinases (p-PI3K), PI3K, phosphorylated protein kinase B (p-Akt), Akt, phosphorylated inhibitor of nuclear factor-κB ( NF-κB)α (p-lκBα) and IκBα protein were detected by Western blotting. The distribution of NF-κB P65 (NF-κB P65) protein in N9 cells was analyzed by immunofluorescence method. Results TREM2 mRNA content in the OGD/R group cells increased significantly within 72 hours after re-oxygenation, and peaked at hour 24 and hour 48. Lenti virus LV- TREM2 effectively promoted the expression of TREM2 mRNA and protein of N9 cells in OGD/R model (P<0.001, P<0.01). Compared with the OGD/R group, the mRNA and protein content of TNF-α, IL-lβ and iNOS decreased significantly, while Arg-1 and IL-10 in OGD/R+LV-TREM2 group increased significantly(P<0.05). Besides, the ratios of P-PI3K/PI3K and p-Akt/Akt increased obviously (P<0.05), the ratio of p-IκBα/IκBα decreased significantly in OGD/ R+ LV-TREM2 group (P<0.001), and the nuclear translocation of NF-κB P65 protein was obviously weakened. Conclusion TREM-2 overexpression exerts anti-inflammatory effect by modulating the polarization of microglia from Ml to M2 type, which is associated with PI3K/Akt and NF-κB signaling pathways regulated by TREM2 in N9 microglia with OGD/R model.

Synergistic Anti-cancer Effect and Mechanism of FZKA Decoction Combined with Gefitinib on A549 Cells / 中国实验方剂学杂志

Objective::To observe the effect of Fuzheng Kangai (FZKA) decoction combined with gefitinib on the cells proliferation, apoptosis, invasion and metastasis of human lung adenocarcinoma A549 cells in vitro and in vivo, and relevant mechanisms. Method::The A549 cell proliferation of the control group, FZKA decoction groups (0.2, 0.4, 0.8, 1.6, 3.2 g·L-1), Gefitinib groups (10, 20, 40, 60, 80, 100 μmol·L-1) for 24, 48, 72 hours, and FZKA decoction (2 g·L-1) combined with Gefitinib (10 μmol·L-1) groups for 24 hours was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The changes of cell apoptosis, invasion and metastasis abilities of A549 cells were analyzed by flow cytometry, Wound Healing, transwell invasion assay. Western blot assay was used to examine the protein expressions of cleaved Caspase-3, B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-2 associated X (Bax), F-box and WD repeat domain-containing (FBW7) and myeloid cell leukemia-1 (MCL-1) in vitro. Result::Compared with control group, FZKA decoction group and Gefitinib group inhibited the cell proliferation, cell apoptosis, cell invasion and metastasis abilities in a dose-dependent and time-dependent manner, and improve the protein expressions of Bax, Caspase-3, FBW7, but decreased the protein expressions of Bcl-2, MCL-1 (P<0.05). Compared with treatment with Gefitinib alone, FZKA combined with Gefitinib inhibited the proliferation of A549 cells, and induced apoptosis more significantly (P<0.05). Compared with treatment with Gefitinib alone, the cell scratch healing and invasion abilities were significantly reduced after combined treatment (P<0.05). FZKA decoction combined with Gefitinib up-regulated Bax, Caspase-3 and FBW7 protein expressions, and down-regulated Bcl-2 and MCL-1 protein expressions compared with treatment with Gifitinib alone (P<0.05). Conclusion::FZKA decoction combined with Gefitinib can inhibit the proliferation, invasion and metastasis, and induce apoptosis on A549 cells. The mechanism may be associated with the FBW7/MCL-1 pathway.

Role and related mechanism of Mst-1 on regulating hypoxic reoxygenation induced autophagy and apoptosis in cardiomyocytes of mouse / 中华心血管病杂志

Objective: To explore the role and related mechanism of mammalian sterile 20-like kinase 1(Mst-1)in regulating hypoxia reoxygenation (HR) induced myocardial cell autophagy and apoptosis. Methods: Enzyme digestion method combined with differential adherent method was used to culture neonatal mouse myocardial cells. HR model was established by hypoxia for 24 hours and reoxygenation for 6 hours. The experimental groups including control group (normal cultured cardiomyocytes), Mst-1 empty virus group (cardiomyocytes transfected with recombinant lentiviral empty vector for 48 hours), Mst-1 knockdown group (recombinant lentivirus carrying Mst-1small interfering RNA (siRNA) was transfected into cardiomyocytes for 48 hours), Mst-1 overexpression group (cardiomyocytes were transfected with recombinant lentivirus carrying Mst-1 gene for 48 hours), HR group (cardiomyocytes exposed to HR), Mst-1 knockdown+HR group (HR model of cardiomyocyte was established 48 hours after transfection with recombinant lentivirus carrying Mst-1siRNA) and Mst-1 overexpression+HR group (HR model of cardiomyocyte was established 48 hours after transfection with recombinant lentivirus carrying Mst-1 gene). Real-time fluorescence quantitative RCR (qPCR) and Western blot were used to detect the relative expression of Mst-1 mRNA and protein in the cells, immunofluorescence staining was used to detect cardiomyocyte troponin T (cTnT), and autophagosomes and autophagy enzyme changes. TUNEL method was used to detect myocardial cell apoptosis, Western blot was adopted to detect autophagy-related protein microtubule-related protein 1 light chain 3 (LC3) Ⅱ/LC3 Ⅰ, P62 and apoptosis-related protein cleaved-caspase 9, pro-caspase 9, cleaved-caspase-3, pro-caspase-3, and myeloid leukemia 1 (MCL-1) expression. MCL-1 inhibitor A1210477 was used to validate the signaling pathway of Mst-1 on regulating cardiomyocyte apoptosis and autophagy. Results: Immunofluorescence detection revealed that the cultured cells expressed cardiomyocyte-specific marker cTnT. The expression of Mst-1 in cardiomyocytes increased in HR model. Lentiviral transfection could effectively inhibit or overexpress Mst-1 in treated cells. The levels of autophagosomes and autophagolysosomes in cardiomyocytes undergoing HR and in Mst-1 overexpression+HR group were lower than those of control group, while autophagosomes and autophagolysosomes in cardiomyocytes of Mst-1 knockdown+HR group was significantly higher than in the HR group (all P<0.05). The TUNEL results showed that the proportion of TUNEL positive cells was significantly increased in the HR group and Mst-1 overexpression+HR group than in the control group, while the proportion of TUNEL positive cells was significantly decreased in the Mst-1 knockdown group+HR group as compared to the HR group (all P<0.05). Western blot results showed that the LC3 Ⅱ/LC3 Ⅰ levels were significantly lower, while the expression levels of P62, cleaved-caspase-9 and cleaved-caspase-3 were significantly higher in the HR group and Mst-1 overexpression+HR group than in control group (all P<0.05). The LC3 Ⅱ/LC3 Ⅰ value was significantly higher, and the expression levels of P62, cleaved-caspase-9 and cleaved-caspase-3 were significantly lower in the Mst-1 knockdown+HR group than in the HR group (P both<0.05). The expression level of P-MCL-1 protein was significantly lower in cardiomyocytes of HR and Mst-1 overexpression+HR group than in control group, and the expression level of P-MCL-1 protein was higher in Mst-1 knockdown+HR group than in HR group (P both<0.05). The recovery experiment showed that inhibiting MCL-1 in cells can block the regulatory effect of Mst-1 siRNA on cell autophagy and apoptosis. Conclusion: Inhibiting Mst-1 expression in cardiomyocytes can promote the autophagy of cardiomyocytes induced by hypoxic reoxygenation and reduce the apoptosis of cardiomyocytes via activating McL-1.

Myeloid cell leukemia-1 protein expression and myeloid cell leukemia-1 gene amplification in non small cell lung cancer

Background: Myeloid cell leukemia-1 (Mcl-1) is a member of the B-cell lymphoma 2 family known to play a significant role in the regulation of apoptosis. Mcl-1 expression has been studied in nonsmall cell lung cancer (NSCLC) cell lines but has not been previously evaluated as a prognostic factor in clinical samples. Materials and Methods: Formalin-fixed, paraffin-embedded sections from 119 NSCLC, including 33 squamous cell carcinomas (SCC), 55 adenocarcinomas (AC), and 31 either pure adenocarcinoma in situ (AIS) or AC with lepidic features were immunostained by an automated method with rabbit polyclonal Mcl-1. Cytoplasmic Mcl-1 (cMcl-1) immunoreactivity was scored based on intensity and percentage of positive tumor cells in both tumor and adjacent benign epithelium in each case. MCL1 amplification was determined by hybrid capture-based comprehensive genomic profiling (CGP) on a separate cohort of 9393 NSCLC samples. Results: Intense diffuse cMcl-1 overexpression was noted in 35/119 (29%) tumors overall and correlated with tumor type (52% AIS vs. 31% AC vs. 6% SCC, P < 0.0001), tumor grade (48% grade 1 vs. 14% grade 2 vs. 31% grade 3, P = 0.007), small tumor size (36% ?3.0 cm vs. 16% >3.0 cm, P = 0.016), and lengthened survival within the AIS subgroup (100% alive vs. 42% expired, P = 0.018) while showing a trend toward correlation with nonrecurrent disease overall (32% nonrecurrent vs. 11% recurrent, P = 0.072) and within the AC subgroup (33% nonrecurrent vs. 0% recurrent, P = 0.092). MCL1 amplification was identified in 569 (6%) of 9393 NSCLC by CGP. Conclusions: cMcl-1 overexpression appears to occur independently from MCL1 gene amplification in NSCLC and correlates with AIS histologic type, lower tumor grade, smaller tumor size, nonrecurrent disease, and increased survival.

To study the correlation of peripheral blood F-box and WD repeat domain-containing domain protein7 and myeloid cell leukemia-1 expression levels with the prognosis of Icotinib administration in elderly patients with advanced non-small-cell lung cancer / 中华老年医学杂志

Objective To study the correlation of the prognosis of Icotinib administration with the expression levels of F-box and WD repeat domain-containing 7(FBW7) and myeloid cell leukemia-1 (MCL-1) in peripheral blood in elderly patients with advanced non-small-cell lung cancer.Methods A total of 76 patients aged 60 years or over diagnosed with non-small-cell lung cancer(NSCLC) with EGFR-sensitive mutations and under Icotinib treatment were enrolled in this study.FBW7 and MCL-1 mRNA expression levels in peripheral blood were detected by real-time quantitative PCR(RT-QPCR).The correlation of FBW7 and MCL-1 expression levels with clinical and histological parameters,overall survival (OS),and progression-free-survival (PFS) was analyzed.Results The FBW7 expression level and the MCL-1 expression level were negative correlated(r =-0.37,P ＜0.001).High FBW7 expression levels and low MCL-1 expression levels in peripheral blood were associated with improved therapeutic efficacy of Icotinib (P＜0.001) and extended OS and PFS.Cox regression analysis showed that the expression levels of FBW7 and MCL-1 in peripheral blood were independent influencing factors for OS and PFS.Conclusions Patients with high FBW7 expression levels and low MCl-1 expression levels are more likely to benefit from Icotinib treatment.Expression levels for either factor can be used as a predictive indicator for the effectiveness of Icotinib and provide guidance for its clinical use.

Value of clearance rate of sTREM-1c in assessing prognosis in patients with septic shock / 实用医学杂志

Objective To monitor the dynamic change of soluble myeloid cells trigger receptor 1 (sTREM-1)and the clearance rate of sTREM-1 (sTREM-1 c) in patients with sepsis shock and to explore its value in assessing the prognosis.Methods A total of 54 patients from January to December 2016 were divided into improved group and death group,sTREM-1 and sTREM-1c level at 1,5,7 and 9 d were monitored and the receiver-operating characteristic curve analysis was used to judge its value in prognosis.Results Comparison of baseline of APACHE Ⅱ score,PCT and age in 2 groups was statistically significant.After treatment,the sTREM-1 level declined,especially in improved group.Similarly,sTREM-1c in improved group at 5,7 and 9 d dropped more significantly than that in death group (P ＜ 0.05).At different time points,sTREM-1 7 topped the predictive value of AUC on the prognosis,followed by APACHE Ⅱ and sTREM-1 5,PCT,sTREM-1 9,sTREM-1c 9 and sTREM-1 1,and sTREM-1c 5 and sTREM-1c 7 were the minimum.Conclusion Effect of dynamic monitoring of sTREM-1 and sTREM-1c analysis in clinic is better than that of simply monitoring of sTREM-1.

Expression changes of soluble urokinase-type plasminogen activator receptor and soluble triggering receptors expressed by myeloid cell-1 in serum of children with primary nephrotic syndrome and their clinical signifi-cance / 中华实用儿科临床杂志

Objective To investigate the expression changes of soluble urokinase-type plasminogen activator receptor(suPAR)and soluble triggering receptors expressed by myeloid cell-1(sTREM-1)in serum of children with primary nephrotic syndrome(PNS)and their clinical significance.Methods A total of 92 cases of newly diag-nosed PNS children were selected in Central Hospital of Yidu Affiliated to Weifang Medical College from June 2014 to September 2016.According to presence or absence of acute tubular necrosis,they were divided into acute renal injury group(27 cases)and non-acute renal injury group(65 cases).According to pathology type,they were divided into mesangial proliferative glomerulonephritis(30 cases),focal segmental glomerulosclerosis(23 cases),membranous ne-phropathy(18 cases),minimal change disease(14 cases)and membrane proliferative glomerulonephritis(7 cases).In the same period,45 healthy children were selected as the healthy control group.The clinical data were collected.The serum levels of suPAR and sTREM-1 were measured by adopting enzyme-linked immunosorbent assay(ELISA). Results The levels of total cholesterol(TC),triglycerides(TG),uric acid(UA),urinary protein/creatinine,24 h urinary protein,urinary N-acetyl-β-glucosaminidase(NAG)and β2-microglobulin(MG)in children with PNS were higher than those in the healthy control group,while serum albumin(ALB)was lower than that in the healthy con-trol group,and the differences were statistically significant(all P<0.05).The serum levels of suPAR and sTREM-1 in PNS patients were(133.09 ± 62.48)ng/L and(79.29 ± 34.68),respectively,which were significantly higher than those in the healthy control group[(31.11 ± 11.61)ng/L and(25.08 ± 8.10)ng/L](t=51.714,49.435;all P=0.000).The serum levels of suPAR and sTREM-1 in acute renal injury group were(188.82 ± 32.21)ng/L and (109.11 ± 24.78)ng/L,respectively,which were significantly higher than those in non -acute renal injury group [(75.96 ± 28.69)ng/L and(52.23 ± 14.07)ng/L]and healthy control group[(31.11 ± 11.61)ng/L and (25.08 ± 8.10)ng/L](F=16 739.607,10 487.256,all P=0.000).The serum levels of suPAR and sTREM-1 in children with focal segmental glomerulosclerosis and membrane proliferative glomerulonephritis were higher than those with minimal change disease,membranous nephropathy and mesangial proliferative glomerulonephritis,and the differences were statistically significant(all P<0.05).Pearson correlation analysis results showed that the serum levels of suPAR and sTREM -1 were positively correlated with TC,TG,urinary protein/creatinine,24 h urinary protein, urinary NAG and β2-MG(all P <0.05),while negatively correlated with ALB(P <0.05). Conclusions The serum levels of suPAR and sTREM-1 are elevated in children with PNS,and which are related with acute renal injury and pathological type,which can reflect the degree of renal tubular disease and kidney function to a certain extent.

Clinical value of serum VEGF,GGT and sTREM-1 in early COPD / 国际检验医学杂志

Objective To explore the clinical value of serum vascular endothelial grow th factor(VEGF), γ-glutamyl transferase(GGT)and soluble myeloid cell trigger receptor-1(sTREM-1)in early chronic obstruc-tive pulmonary disease(COPD).Methods 30 cases of COPD patients admitted from February 2013 to Febru-ary 2015 were selected as the observation group,and 30 healthy subjects were selected as the control group. The levels of serum VEGF,GGT and sTREM-1 were compared between the two groups.Results The levels of serum VEGF,GGT and sTREM-1 were significantly decreased in the observation group after treatment,the difference was statistically significant(P<0.05).The levels of serum VEGF,GGT and sTREM-1 in the obser-vation group were significantly higher than those in the control group(P<0.05).The serum VEGF,GGT and sTREM-1 were respectively detected under the ROC curve area was 0.83,0.80,0.67,specificity and sensitivi-ty were 73.3%,71.1%,58.9% and 91.3%,88.6%,80.2%.ROC curve area of combined detection of VEGF, and GGT in early diagnosis of COPD was 0.94,the sensitivity and specificity were 81.3% and 96.4%;ROC curve area of combined detection of VEGF,and sTREM-1 in early diagnosis of COPD was 0.89,the sensitivity and specificity were 75.5% and 92.8%.Conclusion Serum VEGF,GGT,sTREM-1 level can be used as an important monitoring index for early diagnosis of COPD.

Potential therapeutic target cells of multiple sclerosis-myeloid cells / 药学学报

The discussion on pathophysiological of multiple sclerosis (MS) mainly focuses on T and B cells in adaptive immune response, but less involve in the myeloid cells (dendritic cells, monocytes, macrophages and microglia) in the innate immune system, which also play an important role in the pathogenesis of MS. The myeloid cell population acts as antigen-presenting cells and effector cells in neuroinflammation in the innate immune system. The interactions between T cells and myeloid cells form a vicious cycle which makes the disease continuous deterioration. At present, the studies on the therapeutic drugs for MS mainly are focused on the adaptive immune system, but pay less attention to myeloid cells. In this article, we reviewed the sub-types and functions of myeloid cells, their changes in MS patients and animal models, as well as the effects of some therapeutic drugs for MS on myeloid cells, in the purpose of finding new targets and strategies for development for MS therapy.

Diagnostic value of TREM-1, PCT and CD64 in children with sepsis disease and its effect on the application of low dose hormone / 中国生化药物杂志

Objective To study the diagnostic value of TREM-1,PCT and CD64 in children with sepsis disease and its effect on the application of low dose hormone.Methods 80 cases children with septic disease who received therapy from September 2014 to September 2016 in the third affiliated hospital of wenzhou medical university,28 cases of severe sepsis disease,52 cases of general sepsis disease,and selected 40 cases of healthy children in our hospital during the same period,the levels of TREM-1,PCT,CD64 of three groups were compared.And through the random number table method,those 80 cases children with sepsis disease were divided into the observation group(n=40) and the control group(n=40),the control group was treated with routine treatment,while the observation group was treated with hydroprednisone on the basis of the control group,after treatment for seven days,the levels of TREM-1,PCT,CD64 of two groups were compared.Results The levels of TREM-1, PCT and CD64 in the severe sepsis disease group and general sepsis disease group were higher than the healthy group(P<0.05),and the levels of TREM-1, PCT and CD64 in severe sepsis disease group were significantly higher than the general sepsis disease group(P<0.05).After treatment,the levels of TREM-1,PCT and CD64 were improved in two groups of patients with sepsis disease(P<0.05),after treatment of one day and seven days,the levels of TREM-1,PCT and CD64 in observation group were lower than in the control group(P<0.05).Conclusion TREM-1,PCT,CD64 levels can be used as early detection in children with sepsis disease,low dose glucocorticoid can effectively reduce the expression of TREM-1,PCT and CD64 levels in children with sepsis,it's conducive to the disease control.

PCT and sTREM-1 combined with APACHE Ⅱ score for assessing diagnosis of sepsis and prognostic value / 实用医学杂志

Objective PCT and sTREM-1 combined with APACHE Ⅱ score for assessing diagnosis of sepsis and prognostic value.Methods The clinical data for related patients who had hospitalized between January and October 2015 were collected by prospective study methods.According to China guidelines for treatment of severe sepsis/septic shock (2014).The patients were divided into SIRS group,sepsis group,severe sepsis group,septic shock group,and control group.Results Levels of CRP and sTREM-1 were significantly higher in total sepsis group and subgroups than in SIRS group and control group,with a statistical difference (P ＜ 0.05);but there were no significant differences among he subgroups.Lactic acid level differed statistically between both total sepsis group and its subgroups and both control group and SIRS group,so did level of lactic acid between the other groups and septic shock group (P ＜ 0.05).PCT was significantly higher in total sepsis group and its subgroups than in the control group and SIRS group,so did it in septic shock and severe sepsis group than in sepsis group.PCT level was significantly higher in septic shock than in severe sepsis (P ＜ 0.05).APACHE Ⅱ scores were markedly in total sepsis group and its subgroups than in SIRS group and the control group;and it differed statistically between septic shock group and sepsis group (P ＜ 0.05).According to the ROC curve analysis,the area under the curve was 0.935,0.877,0.816,and 0.856 for PCT,sTREM-1,APACHE Ⅱ,CRP,and lactic acid,respectively.Conclusions Detection of serum PCT and sTREM-1 combined with APACHE Ⅱ score can be used to assess diagnosis of sepsis and prognostic value,which has more benefit to the diagnosis and treatment of sepsis.

Expression and Significance of Mcl-1 Gene in Hepatocellular Carcinoma / 现代检验医学杂志

Objective To investigate the expression and significance of myeloid cell leukemia-1 (Mcl-1 )gene and protein in Hepatocellular Carcinoma(HCC).Methods The expression of Mcl-1 was detected respectively by Reverse Transcription-Polymerase Chain Reaction (RT-PCR),Western blot and ENVISION immunohistochemistry in 20 HCC specimens,19 liver cirrhosis(LC)specimens,and 12 control ones.Their relationship with clinical and pathological characteristics of HCC was investigated.Results The expression of Mcl-1 mRNA in the control group,LC group and HCC group was 0.52±0.17, 3.46±1.7,3.65±2.93,respectively.The level in HCC and LC group was statistically different compared with the control group,respectively (t=7.925,5.334,P<0.05).The relative expression of Mcl-1 protein in LC group (0.51±0.35)and HCC group (0.75±0.36)were significantly higher than that in the control group (0.21±0.19)(t=5.526,6.355,P<0.05).The positive expression rate of Mcl-1 in HCC group was 55.00% (11/20),significantly higher than that in the con-trol group 33.33% (4/12)(t=7.835,P<0.05).The positive expression of Mcl-1 was related to tumor necrosis and TNM staging (χ2=4.201,P<0.05).Conclusion Mcl-1 gene and protein are differentially expressed in HCC,LC and the control, which may be involved in the occurrence,development and malignant transformation of HCC.

Clinical value of inflammatory factors in sepsis / 实用医学杂志

Objective To study the clinical value of sTREM-1, PCT, CRP in sepsis. Methods A prospective study was conducted and clinical data of related patients from January to October 2015 were collected. Thirty-nine cases of sepsis patients , 15 patients with SIRS and 12 normal controls were detected by ELISA then the prognostic value of sTREM-1, PCT, CRP was determined in sepsis. Results In sepsis and SIRS group, sTREM-1 and PCT level were significantly higher than those in control group (P < 0.05); CRP only in sepsis group and control group was statistically significant (P < 0.05). In sepsis group, sTREM-1,PCT and CRP were significantly higher than those in SIRS group (P < 0.05). The ROC curve analysis indicated that sTREM-1,PCT and CRP for SIRS and non SIRS area under the curve was 0.914,0.887 and 0.831 respectively. Conclusion sTREM-1 is a good indicator for early diagnosis of sepsis, and it has high sensitivity and specificity.

The role of soluble triggering receptor expressed on myeloid cell-1 in children with community acquired pneumonia / 临床儿科杂志

Objective To investigate the role of soluble triggering receptor expressed on myeloid cell-l (sTREM-1) in children with community acquired pneumonia.Methods One hundred and seventy-six children with community acquired pneu-monia, 98 cases were mild and 78 cases were servere, were recruited. Thirty healthy children were recruited as control group. The white blood cell count (WBC), neutrophil percentage (N%), C-reactive protein (CRP), procalcitonin (PCT), interleukin-6 (IL-6), interleukin-10 (IL-10), and sTREM-1 were measured.Results The levels of WBC, N%, CRP, IL-6, IL-10, IL-6/IL-10, PCT, and sTREM-1 were signiifcantly different among children with mild pneumonia, severe pneumonia, and healthy controls. All of the indicators were elevated in children with mild and severe pneumonia than those in healthy controls (P<0.05). IL-6/IL-10 sTREM-1 were further signiifcantly elevated in children with severe pneumonia than children with mild pneumonia (P<0.05). IL-6/IL-10 was positively correlated with sTREM-1 (r=0.42,P<0.05).Conclusions sTREM-1 may help for evaluating the severity and outcome of children with community acquired pneumonia.

Construction and identification of Mcl-1 gene shRNA expression plasmid in targeted silenced macrophages / 西安交通大学学报(医学版)

Objective To study the expressions of myeloid cell leukemia-1 (Mcl-1 ) gene in mouse macrophages Raw264.7 and human macrophages THP-1,to screen out the cell lines with high levels of expression as the experimental cells,and based on the screening results to construct the short hairpin RNA(shRNA)eukaryotic expression plasmid targeting mice Mcl-1 gene for transfection and further screen out the shRNA expression plasmid with the most obvious effect in silencing Mcl-1 gene.Methods Semi-quantitative PCR method was used to detect the expression of Mcl-1 mRNA in the two kinds of macrophages.Western blotting was adopted to detect the expressions of Mcl-1 proteins in the two kinds of macrophages.Three different gene loci for Mcl-1 shRNA fragments were designed with small molecules interfering RNA (siRNA)software.Eukaryotic expression plasmid Mcl-1 shRNA 1-3 carrying the shRNA fragments was constructed by a company.And the eukaryotic expression plasmid vector was transfected into scavenger cells Raw264.7 mice via through the liposome.The transfection results 24 h and 48 h after transfection were observed under the inverted fluorescence microscope;Mcl-1 mRNA and protein expression were detected by real time quantitative PCR and Western blot,respectively.Results The relative expression levels of Mcl-1 mRNA and protein in mouse macrophages Raw264.7 were significantly higher than those of human macrophages (P <0.05).The shRNA expression vector constructed within 24 h and 48 h could decrease Mcl-1 mRNA and protein levels in Raw264.7 cells,especially with the most obvious silencing effect at 48 h.The 48-h transfection group differed significantly from normal group,liposome group and negative control group (P <0.05).Compared with Mcl-1shRNA1and Mcl-1shRNA2,Mcl-1shRNA3 had the strongest effect in silencing Mcl-1mRNA and protein.Conclusion We have successfully screened out experimental Raw264.7 cells and Mcl-1 shRNA eukaryotic expression plasmid which has an obvious silencing effect targeting on Mcl-1 in mice macrophages Raw264.7.

Detection of soluble triggering receptor expressed on myeloid cell-1 for diagnosis of early-onset neonatal sepsis / 中华临床感染病杂志

Objective To assess the clinical value of serum soluble triggering receptor expressed on myeloid cell-1 (sTREM-1) for diagnosis of early-onset neonatal sepsis (EONS).Methods A total of 90 neonatal cases with risk factors or symptoms of bacterial infections were enrolled in the study.All infants were admitted to Huai' an Maternity and Child Healthcare Hospital within 24 h after birth during January and June 2014.The enrolled neonates were divided into sepsis group (n =33),general infection group (n =23) and non-infected group (n =34);and the sepsis group was further divided into culture-confirmed group (n =6) and clinical-diagnosed group (n =27).Twenty healthy neonates were also enrolled as the healthy control group.Blood samples were obtained from neonatal patients on d1,d3 and d7 after birth,and for healthy controls,the blood samples were only obtained at the first day.Serum levels of sTREM-1 and interleukin (IL)-6 were measured by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA).The receiver operating characteristic (ROC) curve was applied to evaluate the values of sTREM-1 and IL-6 in diagnosis of EONS.Results Compared with that in general infection group,non-infected group and healthy control group,the serum level of sTREM-1 was significantly higher in sepsis group (all P ＜ 0.05);serum levels of sTREM-1 in general infection group and non-infected group were also higher than that in healthy control group (all P ＜ 0.05);but no difference was observed between general infection group and non-infected group,between culture-confirmed group and clinical-diagnosed group (P ＞ 0.05).Serum level of sTREM-1 showed upward trend during d1-3 after the birth,and downward trend during d3-7.The areas under the ROC curve (AUC) were 0.810 and 0.811 (all P ＜ 0.05) for sTREM-1 levels on d1 and d3 in diagnosis of EONS,and the optimal cut-off values were 234.44 ng/L and 269.79 ng/L,respectively.If sTREM-1 and IL-6 on d1 were combined for diagnosis of EONS,the AUC,sensitivity and specificity were 0.858,92.00％ and 93.10％,respectively.Conclusion Serum level of sTREM-1 in early stage is valuable for diagnosis of EONS,and the combined use of serum sTREM-1 and IL-6 may improve the diagnostic value.

Clinical significance of miR-26a-5p-regulated MCL-1 expression in pre-eclampsia / 中国病理生理杂志

AIM:To investigate the clinical significance of microRNA-26a-5p (miR-26a-5p)-regulated mye-loid cell leukemia-1 (MCL-1) expression in the development of maternal preeclampsia.METHODS:Plasma and placen-tal tissues were collected from 21 cases of normal pregnancy, 13 cases of maternal gestational hypertension, 15 cases of mild preeclampsia and 26 cases of severe preeclampsia.The levels of plasma and placental miR-26a-5p and placental MCL-1 mRNA were detected by real-time PCR.Western blotting analysis was used to determine the protein expression of placen-tal MCL-1.The clinical significance of the above parameters was also analyzed.RESULTS:miR-26a-5p expression gradu-ally increased(P<0.01) in the 4 groups of maternal plasma and placentas with the disease development, and the mRNA expression of MCL-1 was significantly reduced in the placentas (P<0.01), both showing a significant negative correlation (P<0.01).Meanwhile, the expression of miR-26a-5p and MCL-1 protein in the placental tissues was negatively correla-ted (P<0.01).The miR-26a-5p up-regulation in maternal plasma and placental tissues was negatively correlated with ges-tational age, maternal plasma albumin levels and fetal weight, while it was positively correlated with maternal blood pres-sure and urinary protein level (P<0.01), which was in contrary to the down-regulation of placental MCL-1.CONCLU-SION:Up-regulation of miR-26a-5p is involved in the occurrence and development of preeclampsia by down-regulation of MCL-1.