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Objective To explore the predictive value of early changes in platelet counts in the prognosis of severe pneumonia in aged patients.Methods This retrospective study included elderly patients with severe pneumonia,who were ≥65 years old and whose length of ICU stay ≥72 hours,admitted to the intensive care unit(ICU)of NO.2 People's Hospital of Changzhou from January 2014 to January 2017.They were divided into a survival group and a death group according to the 28-day outcome.General information and serum platelet levels at 0,24,36,and 72 hours after admission were collected.Receiver operating characteristic curve (ROC)was plotted according to platelet counts,changes in platelet counts and rates of change in platelet counts to evaluate their predictive value for 28-day prognosis.Kaplan-Meier survival curve was used to analyze the 28-day cumulative survival rate between different groups of patients,who were further divided according to platelet counts at 0 and 72 hours after admission to ICU,changes in platelet counts and rates of change in platelet counts at 72 hours after admission.Results (1)One hundred elderly patients with severe pneumonia were enrolled,among whom 41 cases were in the death group,thus with a mortality of 41.0％.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),sequential organ failure assessment (SOFA),C-reactive protein (CRP),and procalcitonin(PCT)in the death group were significantly higher than those in the survival group.2)Serum platelet levels showed a downward trend in both the death group and the survival group.The level of serum platelets at 72 hours after admission to ICU in the death group was significantly lower than that in the survival group(80.00 × 109/L vs.171.00 × 109/L,Z=5.786,P＜0.05);changes in platelet counts and rates of change in platelet counts in the death group were significantly higher than those in the survival group,especially at 72 hours after admission to ICU(/△PLT72:-79.00 × 109/L vs.-38.00 × 109/L,Z =4.938,P ＜0.05;△PLT 72％:43.6％ vs.-17.7％,Z=6.816,P＜0.05).(3)ROC curve analysis showed that platelet levels,changes in platelet counts,and rates of change in platelet counts could predict 28-day mortality in aged patients with severe pneumonia.The largest area under ROC curve was 0.902 when plotted with the rate of platelet counts at 72 hours after admission to ICU.Using the cut-off value of-36.14％ in the rate of change at 72 hours after admission to evaluate the predictive value in 28 day mortality,the sensitivity and specificity were 89.8％ and 75.6％,respectively.(4)Kaplan-Meier survival analysis showed that the 28 day survival rate was significantly higher and the length of survival was significantly greater when platelet counts at 0 and 72 hours after admission to ICU were higher than the cut-off value,and this also occurred in changes in platelet count and rates of change at 72 hours after admission to ICU.Conclusions Continuous decline in serum p[atelet levels indicates poor prognosis.When combined with platelet counts,changes in platelet counts and rates of change in platelet counts at 72 hours after admission to ICU,it may play an important role in assessing the prognosis of aged patients with severe pneumonia.
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Objective To observe the effect of exosomes secreted by retinal pigment epithelial (RPE) cells which damaged by blue light to Nod-like receptor protein (NLRP3). Methods Cultured ARPE-19 cells were divided into 2 groups; one group of RPE cells were exposed to blue light irradiation for 6 hours, the other group was cultured in routine environment. Total exosomes were extracted from the two groups by differential ultracentrifugation in low-temperature, and examined by transmission electron microscope to identify their forms. The exosomes were then incubated with normal ARPE-19 cells. The expression level of CD63, interleukin (IL)-1β, IL-18 and caspase-1 on the exosome surface were measured by Western blotting. The expressions of NLRP3 mRNA in RPE cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR). Results Blue light damaged the cellular morphology. Transmission electron microscopy showed that the exosomes were 50-200nm in diameter and like double-concave disks. Blue light damaged cell-derived exosomes had significantly higher expression of IL-1β (t=18.04),IL-18 (t=12.55) and caspase-1 (t=14.70) than the control group (P<0.001). ARPE-19 cells cultured with blue light damaged cell-derived exosomes also had significantly higher expression of IL-1β (t=18.59), IL-18 (t=23.95) and caspase-1 (t=35.27) than control exosomes (P<0.001). RT-PCR showed that the relative expression of NLRP3 mRNA of PRE cells in experimental group and control group were 1.000±0.069 and 0.2±0.01, respectively, the difference was significant (t=12.20, P<0.001). Conclusion The expression IL-1β, IL-18 and caspase-1 and NLRP3 mRNA were upregulated by exosomes secreted by blue light damaged-RPE cells.
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Objective To investigate the effects of exosomes from cultured human retinal pigment epithelium (ARPE-19) cells affected by oxidative stress on the proliferation and expression of vascular endothelial growth factor-A (VEGF-A) and Akt of ARPE-19 cells. Methods Culture ARPE-19 cells. The concentration of 2.5μmol/L rotenone was selected to simulate oxidative stress and isolated ARPE-19-exosome. Exosomes were isolated by ExoQuick exosome precipitation solution. Transmission electron microscopy was used to identify the morphology of exosomes. Western blot was used to detect exosomes’ surface-specific maker protein CD63. ARPE-19 cells affected by oxidative stress were cultured with exosome as experimental group, normal ARPE-19 cells were cultured with exosome as control group. The cell proliferation was examined by methyl thiazolyl tetrazolium assay. Western blot and immunofluorescence assay were used to detect the expression levels of VEGF-A and Akt protein. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the levels of VEGF-A mRNA and Akt mRNA. Results The diameter of normal ARPE-19-exosomes ranged from 50 to 150 nm. The isolated exosomes expressed CD63. AREP-19 cells were cultured with ARPE-19 (affected by rotenone)-exosome, the cell viability in experimental group was significantly reduced than in the control group. Green fluorescence was observed in the cytoplasm under fluorescence microscope. Compared with the control group, VEGF-A was up-regulated expressed and Akt was down-regulated expressed. Western blot results showed that, VEGF-A protein expression in the experimental group were higher than the control group. Akt protein expression in the experimental group were less than the control group. The difference was statically significant (t=3.822, 6.527;P<0.05). RT-PCR results showed that VEGF-A mRNA expression levels was higher in the experimental group than the control group. Akt mRNA expression levels was lower in the experimental group than the control group. The difference was statically significant (t=8.805,?7.823;P<0.05). Conclusions Exosomes from ARPE-19 cells affected by oxidative stress inhibit the proliferation of normal ARPE-19 cells, increase the expression of VEGF-A and reduce the expression of Akt.
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Objective To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L norArginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro.Methods The RF/6A cells were divided into the following 4 groups:normal control group (5.0 mmol/L of glucose,group A),high glucose group (25.0 mmol/L,group B),high glucose with 125 mg/L nor-NOHA group (group C),and high glucose with 1％ DMSO group (group D).The proliferation,migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT),transwell chamber and tube assay respectively.The express of Arg Ⅰ,eNOS,iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR),Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells.Results The proliferation,migration,and tube formation ability of group A (t=2.367,5.633,7.045;P＜0.05) and group C (t=5.260,6.952,8.875;P＜0.05)were significantly higher than group B.RT-PCR results showed the Arg Ⅰ and iNOS expression in group B was higher than that in group A (t=6.836,3.342;P＜0.05) and group C (t=4.904,7.192;P＜0.05).The eNOS expression in group B was lower than that in group A and group C (t=4.165,6.594;P＜0.05).ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925,5.368;P＜0.05).IL-1b expression in group B was higher than that in group A and group C (t=5.032,7.792;P＜0.05).Conclusions Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation,migration and tube formation.The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS.
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Objective To explore the predictive value of dynamic serum phosphorus levels in the evaluation of prognosis in patients with sepsis. Methods A retrospective study was conducted. The septic patients admitted to intensive care unit (ICU) of the Second People's Hospital of Changzhou from January 2016 to June 2017 were enrolled, who were ≥18 years old and whose length of ICU stay > 72 hours. These patients were divided into survival group and death group according to 28-day outcome. The general information, the acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ) score within 24 hours of ICU admission, the serum phosphorus at 1, 3, 5, 7 days after admission were collected. Receiver operating characteristic curve (ROC) was plotted according to the dynamic serum phosphorus levels and APACHE Ⅱ score for evaluating the predictive value of 28-day prognosis. Kaplan-Meier survival curve was used to analyze the 28-day cumulative survival rate between two groups of patients, who were divided according to the corresponding time of the serum phosphorus cut-off value. Results ① Eighty-five patients with sepsis were enrolled, among whom 25 cases were in the death group and the mortality was 29.4%. APACHE Ⅱ score in the death group was significantly higher than that in the survival group (22.28±3.98 vs. 16.05±5.44, P < 0.01), the rate of using vasoactive drugs was significantly higher than that in the survival group [64.0% (16/25) vs. 31.7% (19/60), P < 0.01], but there was no significant difference in the length of invasive mechanical ventilation and ICU stay between two groups.② The level of serum phosphorus was increased in survival group along with time of the treatment, and the death group showed a downward trend. The levels of serum phosphorus at 3, 5, 7 days after admission to ICU in death group were significantly lower than those in survival group (mmol/L: 0.90±0.24 vs. 1.05±0.19 at 3 days, 0.96±0.16 vs. 1.11±0.17 at 5 days, 0.83±0.19 vs. 1.21±0.14 at 7 days, all P < 0.01).③ROC curve analysis showed that APACHE Ⅱ score and serum phosphorus level on the 7th day could significantly predict 28-day mortality in patients with sepsis, and the areas under ROC curve (AUC) of them were 0.813 and 0.945 respectively (both P < 0.01). The AUC of serum phosphorus level on the 3rd day and 5th day were 0.692 and 0.745 respectively (both P < 0.01). Based on serum phosphorus cut-off value 1.01 mmol/L on the 7th day to evaluate the predictive value of 28-day mortality, the sensitivity was 91.7%, the specificity was 84.0%, the positive and negative likelihood ratios were 5.73 and 0.10 respectively. ④ Kaplan-Meier survival analysis showed that the 28-day survival rate was significantly higher and the length of survival was significantly longer if the serum phosphorus were higher than the cut-off value at different time points of ICU admission. The 28-day survival rate was significantly higher and the length of survival was significantly longer in the patients with serum phosphorus > 1.01 mmol/L than those serum phosphorus ≤ 1.01 mmol/L on the 7th day [28-day survival rate: 93.2% (55/59) vs. 22.7% (5/22), χ2= 49.697, P = 0.000; survival period (days): 27.1±3.6 vs. 19.8±7.8, t = 4.768, P =0.000]. Conclusion The continuous decline of serum phosphorus indicates poor prognosis, and the serum phosphorus level on the 7th day is one of the most important indicator to evaluate the prognosis of patients with sepsis.
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Objective To observe the effect of exosomes secreted by retinal pigment epithelial (RPE) cells which damaged by blue light to Nod-like receptor protein (NLRP3). Methods Cultured ARPE-19 cells were divided into 2 groups; one group of RPE cells were exposed to blue light irradiation for 6 hours, the other group was cultured in routine environment. Total exosomes were extracted from the two groups by differential ultracentrifugation in low-temperature, and examined by transmission electron microscope to identify their forms. The exosomes were then incubated with normal ARPE-19 cells. The expression level of CD63, interleukin (IL)-1β, IL-18 and caspase-1 on the exosome surface were measured by Western blotting. The expressions of NLRP3 mRNA in RPE cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR). Results Blue light damaged the cellular morphology. Transmission electron microscopy showed that the exosomes were 50-200nm in diameter and like double-concave disks. Blue light damaged cell-derived exosomes had significantly higher expression of IL-1β (t=18.04),IL-18 (t=12.55) and caspase-1 (t=14.70) than the control group (P<0.001). ARPE-19 cells cultured with blue light damaged cell-derived exosomes also had significantly higher expression of IL-1β (t=18.59), IL-18 (t=23.95) and caspase-1 (t=35.27) than control exosomes (P<0.001). RT-PCR showed that the relative expression of NLRP3 mRNA of PRE cells in experimental group and control group were 1.000±0.069 and 0.2±0.01, respectively, the difference was significant (t=12.20, P<0.001). Conclusion The expression IL-1β, IL-18 and caspase-1 and NLRP3 mRNA were upregulated by exosomes secreted by blue light damaged-RPE cells.
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Objective: To assess the regional systolic and diastolic function of left ventricle by pulsed wave Doppler tissue imaging ( PW-DTI) in patients with coronary artery disease ( CAD ) . Methods : Seventy-seven cases of coronary angiography were adopted for study and grouped in terms of severity of coronary arterial lesion. Peak systolic, peak early diastolic, and peak late diastolic motion velocities (Sm, Em and Am, respectively) were recorded at 6 different sites on the basal and medial segments corresponding to the anteroseptal, posteroseptal, lateral, anterior, inferior, and posterior walls of the left ventricle with PW-DTI. Results: Am in segments corresponding to normal coronary arteries (group A) was lower than in segments corresponding to slightly stenosed coronaries (group B). There was no significant difference in Sm and Em between group A and group B. Compared with group A, Sm, Em and Am were significantly decreased in segments corresponding to severely stenosed coronary arteries ( group D). Conclusion : PW-DTI could be used to evaluate quantitatively and accurately the regional mvocardial function of left ventricle in patients with CAD.
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Echocardiography was once clinically considered as a tool for diagnosing coronary artery disease with limited sensitivity and specificity. Recently with the developments of modern electronic technologies,limitations of poor image quality were overcome and several new technical modalities,such as color kinesis(CK),Doppler tissue imaging(DTI)and anatomic M-mode echocardiography(AME)have been demonstrated to be useful for enhancing the accuracy and clinical applicability of the echocardiography. These new noninvasive ultrasonic technologies,adding more direct and quantitative information on specific topics of cardiac function,may play an important role in the study of coronary artery disease.