Objective To research the tourniquet effect on cement mantle thickness in total knee arthroplasty.Methods From June 2013 to June 2014,112 cases of patients were received total knee arthroplasty in the First People's Hospital of Foshan and 94 cases of which received primary operation,82 cases were recruited of the research and randomly divided into experimental group(n=41) without tourniquet and control group(n =41)with tourniquet.The radiological cement mantle thickness was evaluated postoperatively in 2 zones (tibia) on anteroposterior and 4 zones (tibia 2;femur 2) on lateral radiographs,and values were cumulated.Additionally,the calculated blood loss,haemoglobin loss,blood transfusion rate,average transfusion volume,VAS pain score,arc of motion,swelling,ecchymosis and micro thrombus in venules were recorded.Results The study showed that (3.57± 0.62) mm on without tourniquet group and(3.74 ±0.71)mm on tourniquet group in tibia (P=0.240).However,the cement mantle thickness of mm on without tourniquet group(2.00±0.43) mm on tourniquet group(2.19±0.48) in femur (P=0.053),there was no statistically significant difference between two groups.The tourniquet group were reduced on the calculated blood loss,haemoglobin loss,blood transfusion rate and average transfusion volume compared with without tourniquet group(P＜0.05).But VAS pain score,arc of motion,swelling,ecchymosis and micro thrombus in venules were slightly increased in tourniquet group compared with without tourniquet group (P＜0.05).Conclusion The use of a tourniquet in total knee arthroplasty can reduce the calculated blood loss,haemoglobin loss,blood transfusion rate,average transfusion volume,but without using a tourniquet has a better clinical results.

AIM:To investigate the influence of matrine (MA) on the phenotype switching of mouse mono-cytes and alveolar macrophages induced by bleomycin ( BLM) .METHODS:All mice were randomly divided into normal saline (NS) group, BLM group, BLM+NS group and BLM +MA group.The mice were administered with BLM at 2.5 mg/kg via oropharyngeal instillation .The mice in BLM+MA group were treated with MA (15 mg· kg-1 · d-1 ) by oral gavage following BLM administration .The mice were sacrificed on days 3, 7, 14, and 21.The lungs were removed for pathological analysis .The circulating monocyte subsets and polarization state of bronchoalveolar lavage fluid ( BALF)-de-rived alveolar macrophages were analyzed by flow cytometry .RESULTS:The results of HE and Masson trichrome staining in BLM and BLM+NS groups exhibited classical pathological stages of lung fibrosis , including acute inflammation phase and later fibrosis phase .Compared with BLM +NS group, MA treatment alleviated the inflammatory response and the de-gree of fibrosis induced by BLM (P<0.05).There was a rapid change of circulating Ly6Chi monocytes and its magnitude was positively associated with the pulmonary inflammatory response .An expansion of M2-like alveolar macrophages was positively correlated with the magnitude of lung fibrosis .Moreover , MA treatment partially normalized the phenotype switc-hing of monocytes and alveolar macrophages .CONCLUSION:Matrine treatment attenuates BLM-induced pulmonary injury partially via modulating the phenotype switching of monocytes and alveolar mocrophages .

Objective To observe the changes of invasion and migration patterns and integrin expression of malignant glioma cells when Rac and Rho pathways are suppressed,respectively,in 3D hydrogel.Methods Liposome mediated pCMVLifeAct-TagGFP2 plasmids were transfected into glioma U87 cells,and then,these cells were divided into NSC23766 treatment group,Y-27632 treatment group,NSC23766 and Y-27632 combined treatment group,and control group.The three treatment groups were added NSC23766 (100 nmol/mL),Y-27632 (10 nmol/mL),100 nmol/mL NSC23766 and 10 nmol/mL Y-27632,respectively;the cells in the control group were added the same amount of medium.Cells were cultured in hydrogel;the composition of round cells,spindle cells and the mesenchymal and amoeboid cell movement transition were observed under confocal microscopy.The hydrogels of each group were infused into the microslide,and the cell chemotaxis effect were recorded and the cell movement velocities and distances were calculated in the living cell workstation.Immunofluorescence was used to observe the alternation ofintegrin expression.Results U87 cells cultured in 3D hydrogel exhibited spindle-like and round-like shapes,corresponding to mesenchymal and amoeboid cell movement.As compared with that in the control group,the proportion of round cells in the NSC23766 treatment group was significantly higher,and that of spindle cells in the Y-27632 treatment group was significantly higher (P＜0.05).The conversion rate ofmesenchymal-amoeboid transition was 50.0％ in NSC23766 treatment group and amoeboid mesenchymal transition was 42.8％ in Y-27632 treatment group as compared with that in the control group,with significant differences (P＜0.05).The velocity and distance of cells cultured in 3D hydrogel decreased orderly in NSC23766 treatment group,control group,Y-27632 treatment group and combined treatment group in chemotaxis test.The immunofluorescence test showed that integrin expression in the Y-27632 treatment group was significantly higher than that in the other three groups,and that in the control group was statistically higher than that in the NSC23766 treatment group and combined treatment group (P＜0.05).Conclusions 3D hydrogel can be used as a favorable substrate for cell culture.The combination targeted inhabitation of Rac 1 and RohA pathways provides theoretical basis for anti-invasion treatment against glioma.

Objective To explore the protective effcets of Schisandra chinensis extract (SCE) in paraquat (PQ)-induced pulmonary fibrosis in mice ,and its intrinsic molecular mechanisms thereof. Methods A total of 108 mice were randomly allocated into six groups (n=18):control group, model group, low concentration of SCE group (200 mg/kg), medium concentration of SCE group (400 mg/kg), high concentration of SCE group (800 mg/kg) and vitamin C group (100 mg/kg). Except control group, mice were given by intragastric administration with PQ (100 mg/kg) and administered with SCE and Vitamin C once per 24 h after PQ modeling. Mice were sacrificed at 7, 14 and 21 d after modeling. Six mice were executed at different time points. The degree of lung tissue inflammation and fibrosis were observed by HE staining and Masson staining. The mRNA and protein expression levels of transforming growth (TGF)-β1, interleukin (IL)-6 and IL-17 in lung tissue were determined by RT-PCR and ELISA respectively. Results (1) Compared with control group, the lung tissue of model group showed a large number of inflammatory cell infiltration, space congestion, and its inflammation scores increased at 7 and 14 days after modeling (P<0.05). At the same time, compared with model group and vitamin C group, inflammation scores were significantly decreased in medium concentration of SCE group and high concentration of SCE group (P<0.05). (2) Compared with control group, collagen fibers and the degree of fibrosis were significantly increased in model group ,while pulmonary fibrosis were decreased in medium concentration of SCE group and high concentration of SCE group at 14 and 21 days after modeling (P<0.05). (3) With the extension of modeling time, both mRNA and protein expressions of TGF-β1 were obviously elevated, IL-6 decreased and IL-17 reduced after the first increase in PQ group. Compared with PQ group, levels of three cytokines mRNA and protein expression in medium concentration of SCE group and high concentration of SCE group changed as follows:IL-6 level was markedly decreased at 7 and 14 days after modeling;TGF-β1 level was markedly increased at 14 and 21 days after modeling. However, IL-17 level was markedly decrease at three time points(P<0.05). Conclusion SCE can relieve PQ-induced lung inflammation and fibrosis by suppressing TGF-β1, IL-6, and IL-17 expressions.

Objective Pneumosilicosis is characterized by pulmonary fibrosis and cannot be effectively treated at present. This study was to explore the changes of monocyte subsets in the mouse model of silicon dioxide-induced experimental pneumosilicosis and the correlation of the changes with lung inflammatory injury and pulmonary fibrosis. Methods A total of 100 male C57BL/6J mice weighing 18-22 g were equally randomized into a normal saline (NS) group and a silicon dioxide (quartz) group.The model of experimental pneumosilicosis was established by oropharyngeal aspiration of quartz suspension.At 1, 3, 7, 14, and 28 days after treat-ment, the mice were sacrificed and the proportions of different circulating monocyte subpopulations determined by flow cytometry.Dif-ferent types of inflammatory cells in the bronchoalveolar lavage fluid ( BALF) were routinely counted.The inflammation score and col-lagen volume fraction ( CVF) of the lung tissue were obtained by HE and picrosirius red staining. Results At 7 days after quartz treatment, silicotic nodules were observed in the lung tissue.Compared with the NS controls, the model mice showed significantly in-creased inflammation score and CVF at 7 days (0.920 ±0.049 vs 1.400 ±0.089, P<0.01;0.525 ±0.048 vs 1.950 ±0.065, P<0.01) and 28 days (0.800 ±0.089 vs 1.520 ±0.136, P<0.01;0.850 ±0.050 sv 5.300 ±0.776, P<0.01).In comparison with the NS group, the quartz group also exhibited significant increases in the number of total cells at days 1-28 (P<0.01) and the count of neutrophils at days 1-14 (P<0.01) in the bronchoalveolar lavage fluid (BALF) of the model mice, as well as in the number of macrophages in the BALF at 3 days (0.980 ±0.663 vs 6.821 ±2.627, P<0.01), 7 days (1.225 ±0.601 vs 6.697 ±1.864, P<0.01), 14 days (1.492 ±0.438 vs 2.574 ±0.396, P<0.01), and 28 days (2.035 ±0.456 vs 3.249 ±0.492, P<0.01).The count of neutrophilic granulocytes in the BALF was remarkably higher in the quartz than in the NS group at 1, 3, 7, and 14 days (P<0.01) but not at 28 days (P>0.05).Compared with the NS controls, the quartz-treated mice showed markedly increased proportion of Ly6Chimonocytes at all time points, which peaked at 7 days (58.750 ±2.386 vs 78.300 ±2.517, P<0.01), with a positive corre-lation with the inflammation score (P<0.01) and CVF of the lung tissue (P<0.01) at 7 and 28 day. Conclusion The propor-tions of circulating Ly6Chi and Ly6Clo monocytes changed dynamically in the murine model of quartz-induced experimental pneumosilico-sis.The increased proportion of the Ly6Chi monocyte subpopulation might be closely related with lung inflammatory injury and pulmona-ry fibrosis in pneumosilicosis.

Objective To compare the strengths and limitations of the old and revised guidelines for the diagnosis in patients with idiopathic pulmonary fibrosis(IPF).Methods Patients who were admitted and diagnosed as interstitial lung diseases (ILDs) in our hospital from 2009 to 2014 were enrolled in our study.Eachpatient was reevaluated respectively according to the old and revised guidelines of IPF.Results A total of 553 cases were initially reviewed,among whom 355 cases were excluded for pulmonary fibrosis secondary to definite underlying diseases,28 excluded due to high resolution computed tomography(HRCT) not done,26 excluded because serum immunology examination was not available.The remaining 144 cases were finally enrolled in this study including 92 males and 52 females with median age 21-92 (68 ± 11) years old.Twenty five patients (17.4％,25/144) met the diagnostic criteria of IPF by the old guideline.While by the revised guideline,53 patients (36.8％,53/144) were diagnosed as classical IPF,29 patients (20.1％,29/144) as probable cases,another 69 non-IPF patients accounting for 43.1％ (62/144).The result revealed that there's a significant difference between the two guidelines in the diagnosis of IPF.Conclusions The revised guideline favors an early diagnosis of IPF and simplifies the diagnostic process.However the possibility of over diagnosis or missed diagnosis by the revised guideline does exist.On the other hand,despite of the delayed diagnosis by the old guideline,it may reduce the misdiagnosis of IPF in some circumstance.

ObjectiveTo assess the value of the area of ground glass opacities (GGOs) in lungs displayed by high-resolution computed tomography (HRCT) in paraquat (PQ) poisoned patients in evaluating prognosis. Methods Clinical and imaging data of 137 patients with acute PQ poisoning admitted to Affiliated Hospital of the Medical College of Chinese People's Armed Police Forces from January 2012 to August 2014 were analyzed retrospectively. The plasma concentration of PQ on admission and the area of GGOs were compared between two groups. The lung HRCT within 10 days of poisoning was performed every 3 days, and the areas of GGOs were evaluated on five levels, including aortic arch, aortic pulmonary window, left upper lobe bronchial, right inferior pulmonary vein, and left diaphragmatic dome. Receiver operating characteristic curve (ROC) was plotted to evaluate the value of all the parameters for prognosis.Results Among 137 patients, 45 died within 28 days after poisoning, with the mortality rate of 32.85%. The plasma PQ level in the non-survivors was significantly higher than that in the survivors (mg/L:7.06±0.67 vs. 3.51±0.34,t = 5.280,P = 0.000). The areas of GGOs at three time points in the non-survivors were significantly higher than those in the survivors [1-3 days: (32.0±5.0)% vs. (2.5±0.4)%,t = 7.860,P = 0.000;4-6 days: (45.5±5.7)% vs. (2.8±0.5)%,t = 12.420,P = 0.000; 7-10 days: (68.0±4.8)% vs. (3.0±0.6)%, t = 23.950,P = 0.000]. ROC analysis demonstrated that the area under the ROC curve (AUC) of GGOs in 7-10 days was 1.000, which could be used to determine the prognosis, but it was too late for the treatment. The AUC of GGOs in 4-6 days was 0.979, with the threshold of> 12.0%, the specificity of 96.15%, the sensitivity of 85.19%, the positive predictive value of 88.46%, and the negative predictive value of 94.94%, which presented good effect in predicting prognosis in the early stage of acute PQ intoxication. But plasma PQ concentration was relatively poor for determining prognosis, AUC was 0.821, with the threshold of> 1.95 mg/L, the specificity of 34.52%, the sensitivity of 88.64%, the positive predictive value of 41.49%, and the negative predictive value of 85.29%.Conclusions The area of GGOs displayed by HRCT can be used to evaluate the fully developed acute PQ lung injury, and it is superior to plasma PQ concentration. The area of GGOs displayed by HRCT 4-6 days after intoxication can be used for the evaluation of PQ induced pulmonary injury in the early stage and the evaluation of clinical prognosis.

AIM: To establish a cell line of stable silencing of P2X7 receptor (P2X7R) expression through short hairpin RNA ( shRNA)-mediated interference in murine RAW264.7 macrophages, and to investigate the proliferation and apoptosis in the cell line.METHODS:Stable silencing of P2X7 R gene in the RAW264.7 cells was achieved by re-combinant shRNA plasmid targeting murine P2X7 R gene via liposome mediated transfection, followed by G418 selection. The efficacy of plasmid transfection and P2X7 R silencing in G418 resistant cells was verified by immunofluorescent micros-copy and real-time PCR, respectively.The proliferative activity was analyzed by CCK-8 assay and EdU cell proliferation as-say.The cell cycle distribution and apoptosis were evaluated by flow cytometry.RESULTS:The expression of P2X7 R at mRNA and protein levels was down-regulated by 80% in shP2X7 R group compared with negative control ( NC) plasmid transfection.In addition, P2X7 R-silencing cells exhibited higher proliferative activity compared with NC and wild-type RAW264.7 cells (P<0.05).Compared with NC cells, P2X7R silencing resulted in an increase in the phagocytosis of the cells ( P<0.05) .CONCLUSION:A cell line RAW264.7 of stable silencing of P2X7 R expression was successfully es-tablished.P2X7 R gene silencing stimulates the proliferation, and changes phagocytic function in murine RAW264.7 macro-phages.

OBJECTIVETo investigate the prognostic values of initial serum paraquat (PQ) concentration and time lag after PQ ingestion in patients with PQ poisoning.

METHODSA retrospective analysis was performed on 189 patients who ingested PQ between 2.5 and 48 h before admission. The values of initial serum PQ concentration and time lag after PQ ingestion for the prognosis after poisoning were analyzed by the receiver operating characteristic (ROC) curve and multiple logistic regression analysis.

RESULTSThe serum PQ concentration of non-survivors was significantly higher than that of survivors (P<0.01) , and the time lag after PQ ingestion of non-survivors was significantly longer than that of survivors (P<0.01). The ROC curve analysis showed that the area under the ROC curve (AUC) showed no significant difference between initial serum PQ concentration and serum PQ concentration-time data when the time lag after PQ ingestion was between 2.5 and 48 h (z=0.563, P=0.574) ; the AUC showed a significant difference between initial serum PQ concentration and serum PQ concentration-time data when the time lag after PQ ingestion was between 2.5 and 6 h (z=2.199, P=0.034) and between 6 and 48 h (z=2.525, P=0.012).

CONCLUSIONInitial serum PQ concentration has a better predictive power than serum PQ concentration-time data in evaluating the prognosis of patients when the time lag after PQ ingestion is within 6 h. However, serum PQ concentration-time data has a better predictive power than PQ concentration alone in evaluating the prognosis of patients when the time lag after PQ ingestion is between 6 and 48 h.

Area Under Curve ; Humans ; Paraquat ; blood ; Poisoning ; blood ; diagnosis ; Prognosis ; ROC Curve ; Retrospective Studies ; Survivors ; Time Factors

Objective The unbalanced phenotype of pe-ripheral blood monocyte is closely related to the pathological progres-sion of pulmonary fibrosis .The present study was designed to address the dynamic changes of circulating monocyte subsets in the experimen-tal mouse model of pulmonary fibrosis , and explore the relationship of circulating monocyte subsets with pulmonary inflammation and fibro-sis. Methods A total of 100male C 57BL/6J mice were random-ized as control group and a bleomycin A 5 group to be treated with sterile saline and bleomycin A5 at 2 mg/kg, respectively.The mice were sacrificed on day 1, 3, 7, 14, and 21 after treatment.The inflammation score and collagen volume fraction ( CVF) of the lung tissue were obtained by HE and Masson staining .The total number and different types of cells in the bronchoalveolar lavage fluid ( BALF) were counted using the routine method .The mRNA expressions of collagens ⅠandⅢwere determined by real-time PCR, the content of hydroxyproline (HYP) assayed by the chloramine-T method, and the proportions of different monocyte subsets measured by flow cytometry . Results Compared with the saline control , the bleo-mycin A5 group showed significantly increases in the inflammation score at 3 and 7 days ( P<0 .01 ) , CVF at 14 and 21 days ( P<0.01), and the numbers of total cells and macrophages in BALF at 3-21 days, the count of neutrophils granulocytes at 1-3 days (P<0.01), The numbers of neutrophile granulocyles were significant higer than that in control groups on the 1st(9.086 ±1.268 vs 1.108 ±0.229), 3rd(5.551 ±0.511 vs 0.315 ±0.100) and 7th(8.093 ±0.922 vs 0.249 ±0.074)day.The mRNA expressions of collagens ⅠandⅢat 14 and 21 days (P<0.05), the content of HYP at 7-21 days (P<0.01), and the proportion of Ly6Chi mon-ocytes on day 1, which peaked on day 3 (P<0.01) and then decreased from day 14 to 21.The proportion of Ly6Chi monocytes was positively correlated with the inflammation score (P<0.000 1) and CVF of the lung tissue (P=0.001 3). Conclusion In the mouse model of bleomycin A5-induced pulmonary fibrosis, dynamic changes of circulating Ly6Chi and Ly6Clo monocyte subsets occurred in different pathophysiological stages .Compared with the pathological process of inflammatory infiltration , Ly6Chi circulating monocytes displayed a rapid response to tissue injury and inflammation .The increased proportion of Ly6Chi monocyte subsets might be closely re-lated with pulmonary inflammation and fibrosis .