Effect of proprotein convertase subtilisin/kexin type 9 on platelet activation associated with sepsis / 中华危重病急救医学

Objective:To investigate the effect of proprotein convertase subtilisin/kexin type 9 (PCSK9) on platelet activation in sepsis.Methods:① Clinical trial: a prospective study was conducted. Patients with sepsis and septic shock aged ≥ 18 years old who met the diagnostic criteria of Sepsis-3 admitted to the department of intensive care medicine of the Affiliated Hospital of Binzhou Medical College from January to October in 2021 were selected as subjects. Healthy subjects in the same period were taken as healthy control group. Platelet count (PLT) in the first routine blood test after admission was recorded. Venous blood was taken 1 day after diagnosis, and serum PCSK9 level was determined by enzyme-linked immunosorbent assay (ELISA). The differences of PCSK9 level and PLT between the two groups were compared, and subgroup analysis was conducted based on PLT for patients with sepsis. The correlation between PCSK9 level and PLT in septic patients was analyzed by Pearson correlation method. ② Animal experiment: 80 male C57BL/6 mice were randomly divided into control group, sepsis model group [lipopolysaccharide (LPS) group], PCSK9 inhibitor pretreatment group (PCSK9 inhibitor+LPS group) and PCSK9 inhibitor control group (PCSK9 inhibitor group), with 20 mice in each group. The mouse model of sepsis was reproduced by intraperitoneal injection of LPS 12 mg/kg, and the control group and PCSK9 inhibitor group were intraperitoneally injected with the same amount of sterile normal saline. PCSK9 inhibitor+LPS group and PCSK9 inhibitor group were pretreated with PCSK9 inhibitor 5 mg/kg intraperitoneal injection for 7 days before injection of LPS or normal saline, respectively, and the control group and LPS group were injected with an equal amount of sterile normal saline. The lung tissues were taken for pathological and immunohistochemical observation 24 hours after modeling. Blood was taken from the heart for determining PLT. Platelet activation was detected by flow cytometry. The expression level of platelet-activation marker CD40L was detected by Western blotting.Results:① Clinical trial: there were 57 cases in the sepsis group and 27 cases in the healthy control group. Serum PCSK9 level in the sepsis group was significantly higher than that in the healthy control group (μg/L: 232.25±72.21 vs. 191.72±54.92, P < 0.05), and PLT was significantly lower than that in the healthy control group [×10 9/L: 146.00 (75.50, 204.50) vs. 224.00 (194.00, 247.00), P < 0.01]. Subgroup analysis showed that the serum PCSK9 level in the thrombocytopenia patients ( n = 20) was significantly higher than that in the non-thrombocytopenia patients ( n = 37; μg/L: 264.04±60.40 vs. 215.06±72.95, P < 0.01). Correlation analysis showed a significant negative correlation between serum PCSK9 levels and PLT in septic patients ( r = -0.340, P = 0.010). ② Animal experiment: there were no significant pathological changes in lung tissue in the control group and PCSK9 inhibitor group under light microscope, and no significant differences in PLT, platelet activation and plasma CD40L protein expression was found between the two groups. In the LPS group, a large number of inflammatory cells were infiltrated in the pulmonary interstitium, the alveolar structure was damaged obviously, the alveolar septum was widened, the alveolar cavity was extensively bleeding, the capillary dilatation with bleeding and platelet aggregation were found, the PLT was significantly decreased, the platelet activation and the expression level of CD40L protein in plasma were significantly increased. The infiltration of inflammatory cells in lung tissue of mice in the PCSK9 inhibitor+LPS group was reduced to a certain extent, the thickening of alveolar septa was reduced, the platelet aggregation in lung tissue was decreased as compared with the LPS group, the PLT was significantly increased (×10 9/L: 515.83±46.60 vs. 324.83±46.31, P < 0.05), the platelet activation and the expression level of CD40L protein in plasma were significantly decreased [positive expression rate of platelet activation dependent granule surface facial mask protein CD62P: (12.15±1.39)% vs. (18.33±2.74)%, CD40L protein (CD40L/β-actin): 0.77±0.08 vs. 1.18±0.10, both P < 0.05]. Conclusion:PCSK9 level has a certain effect on promoting platelet activation in sepsis, and inhibition of PCSK9 level may have potential research value in improving adverse outcomes caused by sepsis thrombocytopenia.

Analysis of chest imaging features of novel coronavirus pneumonia, bacterial pneumonia and viral pneumonia / 中华危重病急救医学

Objective:To investigate and summarize the chest CT imaging features of patients with novel coronavirus pneumonia (COVID-19), bacterial pneumonia and other viral pneumonia.Methods:Chest CT data of 102 patients with pulmonary infection due to different etiologies were retrospectively analyzed, including 36 patients with COVID-19 admitted to Hainan Provincial People's Hospital and the Second Affiliated Hospital of Hainan Medical University from December 2019 to March 2020, 16 patients with other viral pneumonia admitted to Hainan Provincial People's Hospital from January 2018 to February 2020, and 50 patients with bacterial pneumonia admitted to Haikou Affiliated Hospital of Central South University Xiangya School of Medicine from April 2018 to May 2020. Two senior radiologists and two senior intensive care physicians were participated to evaluated the extent of lesions involvement and imaging features of the first chest CT after the onset of the disease.Results:Bilateral pulmonary lesions were more common in patients with COVID-19 and other viral pneumonia, and the incidence was significantly higher than that of bacterial pneumonia (91.6%, 75.0% vs. 26.0%, P < 0.05). Compared with other viral pneumonia and COVID-19, bacterial pneumonia was mainly characterized by single-lung and multi-lobed lesion (62.0% vs. 18.8%, 5.6%, P < 0.05), accompanied by pleural effusion and lymph node enlargement. The proportion of ground-glass opacity in the lung tissues of patients with COVID-19 was 97.2%, that of patients with other viral pneumonia was 56.2%, and that of patients with bacterial pneumonia was only 2.0% ( P < 0.05). The incidence rate of lung tissue consolidation (25.0%, 12.5%), air bronchial sign (13.9%, 6.2%) and pleural effusion (16.7%, 37.5%) in patients with COVID-19 and other viral pneumonia were significantly lower than those in patients with bacterial pneumonia (62.0%, 32.0%, 60.0%, all P < 0.05), paving stone sign (22.2%, 37.5%), fine mesh sign (38.9%, 31.2%), halo sign(11.1%, 25.0%), ground-glass opacity with interlobular septal thickening (30.6%, 37.5%), bilateral patchy pattern/rope shadow (80.6%, 50.0%) etc. were significantly higher than those of bacterial pneumonia (2.0%, 4.0%, 2.0%, 0%, 22.0%, all P < 0.05). The incidence of local patchy shadow in patients with COVID-19 was only 8.3%, significantly lower than that in patients with other viral pneumonia and bacterial pneumonia (8.3% vs. 68.8%, 50.0%, P < 0.05). There was no significant difference in the incidence of peripheral vascular shadow thickening in patients with COVID-19, other viral pneumonia and bacterial pneumonia (27.8%, 12.5%, 30.0%, P > 0.05). Conclusions:The probability of ground-glass opacity, paving stone and grid shadow in chest CT of patients with COVID-19 was significantly higher than those of bacterial pneumonia, and it was more common in the lower lungs and lateral dorsal segment. In other patients with viral pneumonia, ground-glass opacity was distributed in both upper and lower lungs. Bacterial pneumonia is usually characterized by single lung consolidation, distributed in lobules or large lobes and accompanied by pleural effusion.

Mechanism of high mobility group protein B1 in lipopolysaccharide-induced acute lung injury/acute respiratory distress syndrome / 中华危重病急救医学

Objective:To investigate the role and possible pathogenesis of high mobility group protein B1 (HMGB1) in lipopolysaccharide (LPS)-induced acute lung injury/acute respiratory distress syndrome (ALI/ARDS).Methods:① In vivo, 24 SPFC57BL/6 male mice were randomly divided into normal control group, ALI/ARDS model group, ethyl pyruvate (EP) treatment group and EP control group, with 6 mice in each group. The ALI/ARDS model was established by intraperitoneal injection of 20 mg/kg LPS. Mice in normal control group and EP control group were intraperitoneally injected with the same amount of sterile normal saline. Then, mice in the EP treatment group and EP control group were intraperitoneally injected with 40 mg/kg HMGB1 inhibitor EP. After 6 hours, the mice were sacrificed and lung tissues were collected. The expressions of heparan sulfate (HS), syndecans-1 (SDC-1), heparanase (HPA) and matrix metalloproteinases-9 (MMP-9) in lung tissues were detected by immunofluorescence technique. Orbital blood of mice was collected and serum was extracted to detect the content of HMGB1 by enzyme linked immunosorbent assay (ELISA). ② In vitro, human umbilical vein endothelial cells (HUVECs) were randomly divided into 6 groups: normal control group, HUVECs damage group (treated with 1 mg/L LPS for 6 hours), HMGB1 group (treated with 1 μmol/L recombinant HMGB1 for 6 hours), HMGB1+EP group (treated with recombinant HMGB1 for 1 hour and then added 1 μmol/L EP for 6 hours), LPS+EP group (treated with LPS for 1 hour and then added 1 μmol/L EP for 6 hours), EP group (treated with 1 μmol/L EP for 6 hours). The expressions of HS, SDC-1, HPA and MMP-9 in endothelial cells were detected by immunofluorescence technique. Results:① In vivo, light microscopy showed that the alveolar space was thickened after LPS stimulation, and there were a large number of inflammatory cells infiltrating in the alveolar space. Compared with ALI/ARDS model group, the expressions of HS and SDC-1 in lung tissue of EP treatment group were significantly increased [HS (fluorescence intensity): 0.80±0.20 vs. 0.53±0.02, SDC-1 (fluorescence intensity): 0.72±0.02 vs. 0.51±0.01, both P < 0.05], and the expressions of HPA and MMP-9 were significantly decreased [HPA (fluorescence intensity): 2.36±0.05 vs. 3.00±0.04, MMP-9 (fluorescence intensity): 2.55±0.13 vs. 3.26±0.05, both P < 0.05]; there were no significant changes of the above indexes in EP control group. Compared with ALI/ARDS model group, the content of serum HMGB1 in EP treatment group decreased significantly (μg/L: 131.88±16.67 vs. 341.13±22.47, P < 0.05); there was no significant change in the EP control group. ② In vitro, compared with HMGB1 group, the expressions of HS and SDC-1 in HMGB1+EP group were significantly higher [HS (fluorescence intensity): 0.83±0.07 vs. 0.56±0.03, SDC-1 (fluorescence intensity): 0.80±0.01 vs. 0.61±0.01, both P < 0.05], and the expressions of HPA and MMP-9 were significantly lower [HPA (fluorescence intensity): 1.30±0.02 vs. 2.29±0.05, MMP-9 (fluorescence intensity): 1.55±0.04 vs. 2.50±0.06, both P < 0.05]; the expression of HS, SDC-1, HPA and MMP-9 had no significant changes in EP group. Conclusion:HMGB1 participates in LPS-induced injury of endothelial cell glycocalyx, leading to increased lung permeability, and inhibition of HMGB1 can alleviate lung injury.

Prognostic value of PCSK9 and blood lipid in patients with sepsis / 中华危重病急救医学

Objective:To investigate the prognostic value of proprotein convertase subtilisin/kexin type 9 (PCSK9) and blood lipid indexes in patients with sepsis.Methods:Patients with sepsis or septic shock who were ≥ 18 years old and met the Sepsis-3.0 diagnostic criteria admitted to the department of critical care medicine of Binzhou Medical University Hospital from January to October 2021 were enrolled. Healthy adults at the same period were selected as healthy control group. Baseline characteristics, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score were recorded. Venous blood samples were collected within 24 hours after diagnosis, and serum PCSK9 was determined by enzyme-linked immunosorbent assay (ELISA) at 1, 3 days and 5 days. Meanwhile, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglyceride (TG) and lipoprotein A were detected. The differences of each index between sepsis group (28-day death group and survival group) and healthy control group were compared. Meanwhile, the indexes of patients with different severity and 28-day prognosis in sepsis group were compared. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of PCSK9 and blood lipid for the prognosis of sepsis. Multivariate Logistic regression was used to analyze the influencing factors for the prognosis of sepsis, and the Kaplan-Meier survival curve at 28th day was drawn.Results:There were 50 patients in sepsis group (including 19 patients with sepsis, 31 patients with septic shock) and 27 patients in healthy control group. In the sepsis group, 19 patients died and 31 patients survived within 28 days. The serum PCSK9 in the sepsis group was significantly higher than that in the healthy control group [μg/L: 223.09 (198.47, 250.82) vs. 188.00 (165.27, 214.90), P < 0.01], and HDL-C, LDL-C, TC and lipoprotein A were significantly lower than those in the healthy control group [HDL-C (mmol/L): 0.82±0.35 vs. 1.45±0.24, LDL-C (mmol/L): 1.53 (1.14, 2.47) vs. 2.89 (2.55, 3.19), TC (mmol/L): 2.03 (1.39, 2.84) vs. 4.24 (3.90, 4.71), lipoprotein A (g/L): 8.80 (5.66, 17.56) vs. 27.03 (14.79, 27.03), all P < 0.01]. PCSK9 in the sepsis death group was higher than that in the survival group [μg/L: 249.58 (214.90, 315.77) vs. 207.01 (181.50, 244.95), P < 0.01], and the HDL-C, LDL-C and TC were lower than those in the survival group [HDL-C (mmol/L): 0.64±0.35 vs. 0.93±0.30, LDL-C (mmol/L): 1.32±0.64 vs. 2.08±0.94, TC (mmol/L): 1.39 (1.01, 2.23) vs. 2.69 (1.72, 3.81), all P < 0.01]. With the progression of the disease, the PCSK9 in the sepsis death group and the survival group was significantly lower than that within 1 day of diagnosis (all P < 0.05). ROC curve analysis showed that PCSK9 had higher predictive value of 28-day death than HDL-C, LDL-C, TC [area under ROC curve (AUC) and 95% confidence interval (95% CI): 0.748 (0.611-0.885) vs. 0.710 (0.552-0.868), 0.721 (0.575-0.867), 0.702 (0.550-0.854)]. Multivariate Logistic regression analysis showed that PCSK9 was an independent risk factor affecting the 28-day prognosis of sepsis (β value was 1.014, P = 0.020). Kaplan-Meier survival curve analysis showed that when PCSK9 ≥ 208.97 μg/L, with the increase of PCSK9, the 28-day survival rate of sepsis patients decreased significantly. Conclusions:PCSK9, HDL-C, LDL-C and TC can all predict the 28-day prognosis of patients with sepsis. The prognostic value of PCSK9 is the highest. PCSK9 is an independent risk factor affecting the prognosis of sepsis. In the early stage of the disease, PCSK9 may have a good predictive value for the prognosis of sepsis. When PCSK9 ≥ 208.97 μg/L, the 28-day survival rate decreased significantly.

Assistant role of manifestations under tracheoscopy in the diagnosis of invasive pulmonary aspergillosis in severe patients / 中华危重病急救医学

Objective:To evaluate the assistant role of manifestations under tracheoscopy in the diagnosis of invasive pulmonary aspergillosis (IPA) in severe patients.Methods:A retrospective study was conducted. The patients with suspected IPA admitted to intensive care unit (ICU) of Affiliated Hospital of Binzhou Medical College from January 2015 to December 2019 were enrolled. The diagnosis, clinical diagnosis and suspected diagnosis were made according to the grading criteria of Guidelines for the diagnosis and treatment of invasive fungal infection in severe patients (2007). Those who met the criteria were enrolled in the IPA group, and those who did not meet the criteria or other pathogens were enrolled in the non-IPA group. The general data of the patients were collected, and the changes of tracheal and bronchial mucosa under tracheal microscope before and after treatment were recorded, as well as the results of galactomannan (GM) test and aetiology culture of bronchoalveolar lavage fluid (BALF). The baseline, bronchoscopy and pulmonary CT manifestations and their dynamic changes were compared in each group. Results:A total of 142 patients with suspected IPA were finally enrolled. Among them, 12 were pathologically proven IPA, 77 were probable IPA, 22 were possible IPA, and 31 were undefined IPA. Of the 142 patients, 60 had typical manifestations of mucosal injury under bronchoscopy, including 7 proven IPA patients (58.3%), 52 probable IPA patients (67.5%), and 1 possible IPA patient (4.5%), but none undefined IPA patient. The patients undergoing lung CT scan were 12 proven IPA patients (100%), 73 probable IPA patients (94.8%), and 21 possible IPA patients (95.5%), respectively. Most of the Chest CT showed patchy or strip density increasing and other non-specific manifestations. There were 3 proven IPA patients (25.0%), 7 probable IPA patients (9.0%), and 0 possible IPA patient (0%) who had typical IPA CT manifestations (halo sign and cavity or crescent sign). Among the patients of proven IPA and probable IPA (89 cases), there were a total of 35 cases with endoscopic airway mucosal injury and tracheoscopy reexamination ≥ 3 times. All the 35 patients received anti-aspergillus treatment, among which 16 survived and 19 died. Among the 16 patients who survived, the microscopic appearance of mucosal injury was gradually reduced and the clinical manifestations were gradually improved. Of the 19 patients who died, 16 had deteriorated endoscopic airway mucosal injury.Conclusions:The specific manifestations of severe patients with bronchial mucosal injury are of great significance in the diagnosis of IPA. In the case of severe patients who cannot receive pathological examination or chest CT in time, dynamic observation of the changes of airway mucosal injury is a simple auxiliary method to discover the changes of patients' condition in time, evaluate the effect of antifungal therapy and the prognosis of IPA.

Prognostic analysis of severe patients with bloodstream infection caused by Enterobacteriaceae bacteria / 中华危重病急救医学

Objective:To investigate the prognostic factors of severe patients with bloodstream infection (BSI) caused by Enterobacteriaceae bacteria. Methods:Patients suffered from BSI caused by Enterobacteriaceae bacteria admitted to department of critical care medicine of Binzhou Medical University Hospital from October 2016 to October 2019 were enrolled. The information of gender, age, combined shock, acute physiology and chronic health evaluationⅡ(APACHEⅡ), sequential organ failure assessment (SOFA), sensitivity of initial antibiotics, as well as the baseline of procalcitonin (PCT), white blood cell count (WBC), platelet (PLT), albumin (ALB) were collected. The 72-hour PCT clearance rate (72 h PCTc) was calculated after 72 hours' treatment. According to the clinical outcome after 28 days, the patients were divided into recovery group and death group. The differences of clinical indicators between the two groups were compared, and then the statistical significant variables were further performed by Logistic regression to analyze the factors affecting the prognosis of patients. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive efficacy of the factors in severe BSI. Results:A total of 86 patients were enrolled, among whom 54 cases recovered while 32 cases died, and the 28-day mortality was 37.2%. There was no significant difference in gender, age, sensitivity of initial antibiotics, baseline levels of PCT and WBC between two groups. In the death group, the shock incidence, APACHEⅡscore, SOFA score were significantly higher than those in recovery group [shock incidence: 84.4% (27/32) vs. 46.3% (25/54), APACHEⅡ: 24.94±7.65 vs. 17.02±6.57, SOFA: 11.00±3.27 vs. 6.30±2.65, all P < 0.01]; the PLT and ALB baseline levels, 72 h PCTc were significantly lower than those in recovery group [PLT (×10 9/L): 73.38±49.15 vs. 138.69±101.80, ALB (g/L): 25.47±5.91 vs. 28.59±4.53, 72 h PCTc: -44 (-170, 27)% vs. 63 (40, 77)%, all P < 0.01]. The above 6 variables were included in Logistic regression. The results showed that SOFA score was a risk factor for death in these patients [odds ratio ( OR) = 1.930, P = 0.037], while 72 h PCTc and ALB were protective factors ( OR values were 0.043, 0.783, P values were 0.008, 0.047). The SOFA, 72 h PCTc and ALB can be used to predict the prognosis of severe BSI, and the diagnostic value of the combination of three factors was the largest [area under the ROC curve (AUC) = 0.953, 95% confidence interval (95% CI) was 0.909-0.997], the sensitivity was 100%, and the specificity was 79.6%. Conclusions:Severe patients with BSI caused by Enterobacteriaceae bacteria had a high mortality. Higher SOFA score, and lower ALB and 72 h PCTc predicted the adverse outcome. The combination of the three factors has the greatest prognostic efficacy.

Study on Mechanism of Platelet Aggregation Inhibitory Effects and Lung Tissue Protective Effects of Baicalein in Model Rats with Acute Pulmonary Embolism Based on Notch Signaling Pathway / 中国药房

OBJECTIVE：To explore the mec hanism of baicalein plat elet aggregation inhibitiory effect and lung tissue protective effect of baicalein in model rats with acute pulmonary embolism. METHODS ：Totally 36 rats were randomly divided into normal control group （n＝6）and modeling group （n＝30）. The acute pulmonary embolism model was established by autologous thrombus replication in modeling group ，and the sham operation of rats in normal control group was carried out. After modeling ， 30 model rats were randomly divided into model control group ，positive drug group （low molecular weight heparin calcium 0.01 mL/kg，subcutaneous injection ），baicalein low-dose ，middle-dose and high-dose groups （25，50，100 mg/kg，intraperitoneal injection），with 6 rats in each group. Normal control group and model control group were intraperitoneally injected constant volume of normal saline ；administration groups were given relevant medicine ，once a day ，for consecutive 7 d. After medication ， platelet aggregation rates of rats after activated with adenosine diphosphate （ADP） and arachidonic acid （AA） and platelet activation index （RPI）were detected ；lung histopathology was observed by HE staining ；serum platelet activation markers granule membrane（CD62P）and lysosomal membrane glycoprotein （CD63），growth differentiation factor- 15（GDF-15）and N-terminal B-type natriuretic peptide （NT-proBNP）were measured by ELISA. The mRNA expression levels of Notch 2，Notch3 and Notch signaling ligand PLL 1，JAG2 were detected by RT-PCR method. The protein expression levels of Notch 2，Notch3，DLL1 and JAG2 in lung tissue were detected by immunohistochemistry and Western blotting assay. RESULTS ：Compared with normal control group，plasma ADP-activated platelet aggregation rate ，AA-activated platelet aggregation rate ，RPI，serum levels of CD 62P， CD63，GDF-15 and NT-proBNP were increased significantly （P＜0.05）. The lung tissue of rats was in a state of severe inflammatory infiltration. mRNA and protein expression levels of Notch 2，Notch3，DLL1 and JAG 2 in lung tissue decreased significantly（P＜0.05）. Compared with model control group ，changes of above indexes of rats were improved significantly in baicalein groups （P＜0.05）. CONCLUSIONS ：Baicalein can reduce platelet aggregation and improve the pathological state of lung tissue in rats with acute pulmonary embolism. Its mechanism 0270） may be related to activating Notch signal pathway.

Practice of constructing a performance evaluation system of teaching faculty in teaching management of municipal general hospitals / 中华医学教育探索杂志

The construction of teaching faculty is not only a core element to carry out medical education in municipal general hospitals, but also a weak link. Constructing a performance evaluation system of teaching faculty has important significance in promoting teaching reform and innovation, and improving teachers' enthusiasm for teaching as well as their teaching quality in municipal general hospitals. Taking the Affiliated Dongguan People's Hospital of Southern Medical University as an example, this paper focused on the construction of teaching faculty and aimed to provide references for municipal general hospitals to build a performance evaluation system through the promotion of a series of measures such as teaching ability assessment, quantitative assessment of teaching work, teaching quality evaluation, and teaching contribution assessment.

Practice of constructing a performance evaluation system of teaching faculty in teaching management of municipal general hospitals / 中华医学教育探索杂志

The construction of teaching faculty is not only a core element to carry out medical education in municipal general hospitals,but also a weak link.Constructing a performance evaluation system of teaching faculty has important significance in promoting teaching reform and innovation,and improving teachers' enthusiasm for teaching as well as their teaching quality in municipal general hospitals.Taking the Affiliated Dongguan People's Hospital of Southern Medical University as an example,this paper focused on the construction of teaching faculty and aimed to provide references for municipal general hospitals to build a performance evaluation system through the promotion of a series of measures such as teaching ability assessment,quantitative assessment of teaching work,teaching quality evaluation,and teaching contribution assessment.

Efficacy of bundle care under the supervision of professionals in preventing ventilator-associated pneumonia / 中国感染控制杂志

Objective To investigate the efficacy of bundle care under the supervision of professionals in preven-ting ventilator-associated pneumonia(VAP). Methods Patients who received mechanical ventilation between July 2013 and June 2014 in the intensive care unit(ICU)of a hospital were selected,July-December 2013 was as control group (n= 150),patients received bundle care,but without special supervision;January-June 2014 was as special supervision group (n= 177),patients received bundle care under the supervision of professionals,incidence of VAP,duration of mechanical ventilation,and length of ICU stay were compared between two groups. Results In special supervision group,141 patients received invasive mechanical ventilation,mechanical ventilation day were 1937 days,9 patients developed VAP,incidence of VAP was 4.65‰ ;in control group,127 patients received inva-sive mechanical ventilation,mechanical ventilation day were 1965 days,21 patients developed VAP,incidence of VAP was 10.69‰ ,difference was statistically significant between two groups(χ2= 5.68,P= 0.042). In special su-pervision group,overall compliance rate of bundle management strategies,duration of average invasive mechanical ventilation,and length of ICU stay were 94.92% ,(11.11±2.57)days,and(15.11±2.88)days respectively,in con-trol group were 48.67% ,(14.67±4.35)days,and(19.33±5.81)days respectively,difference was statistically sig-nificant between two groups (all P<0.05). Conclusion Bundle care strategies supervised by professionals can ef-fectively prevent the occurrence of VAP,the implementation of bundle care strategies should be set up according to the clinical practice.

Variation and clinical value of endothelial glycocalyx in the patients with septic shock / 中华危重病急救医学

Objective To explore the variation and clinical value of the degradation of endothelial glycocalyx in the patients with septic shock. Methods A prospective case control study was conducted. Patients of 18 years or older diagnosed with septic shock and admitted to Department of Critical Care Medicine of Affiliated Hospital of Binzhou Medical University from June 2014 to May 2015 were enrolled. The levels of degradation products, including hyaluronic acid (HA) and heparin sulfate (HS), at 0, 6, 12, 24, 48 hours were determined, while 20 healthy people were enrolled and served as controls. The changes of HA and HS were analyzed in the patients with septic shock. The differences of HA and HS between survival group and death group after 28 days were also analyzed. The relationships between HA, HS and tumor necrosis factor-α (TNF-α), sequential organ failure assessment (SOFA) score, arterial blood lactate (Lac), platelet, albumin were analyzed by Pearson correlation analysis. The receiver-operating characteristic (ROC) curve was plotted to assess the prognostic value of HA and HS for patients with septic shock. Results Thirty-one patients diagnosed as septic shock were enrolled, among whom 17 patients died after 28 days, with a mortality of 54.8%. The levels of HA and HS in patients with septic shock were increased significantly as compared with those of health control group, peaked at 48 hours, and the levels of HA and HS at 48 hours were significantly higher than those at 0 hour [HA (μg/L): 119.47±32.44 vs. 94.84±23.63, HS (μg/L): 72.83±19.03 vs. 58.83±16.63, both P < 0.05]. The levels of HA and HS at 0 hour and 48 hours in death group were significantly higher than those of the survival group [HA (μg/L): 130.42±27.67 vs. 93.29±29.80, 105.14±19.18 vs. 70.82±13.24; HS (μg/L): 67.23±25.01 vs. 39.23±14.58, 79.74±19.84 vs. 56.17±14.53, all P < 0.05]. The levels of HA and HS in patients with septic shock were remarkably positively correlated with the levels of TNF-α, SOFA score, Lac, and platelet, but were remarkably negatively correlated with albumin levels (r value of HA was 0.595, 0.462, 0.545, 0.466, -0.534, respectively; r value of HS was 0.607, 0.468, 0.563, 0.547, -0.455, respectively; all P < 0.05). It was demonstrated by ROC curves that the areas under ROC curve (AUC) of HA and HS at 0 hour and 48 hours for predicting the prognosis of patients with septic shock were 0.881, 0.940 and 0.833, 0.821, respectively, the sensitivities of HA and HS were 87.5%, 100.0% and 83.3%, 81.3%, respectively, and the specificities of HA and HS were 82.6%, 78.3% and 91.3%, 78.3%, respectively. Conclusions The concentrations of degradation products generated by endothelial glycocalyx in the blood of the patients with septic shock are remarkably increased. The elevated levels of the degradation products are closely associated with the severity of septic shock, microcirculation disturbance, and the levels of inflammatory factors.

Analysis of the distribution characteristics and drug resistance of non-fermenting bacterial infection in intensive care unit from 2009 to 2015 / 中华危重病急救医学

Objective To study the distribution characteristics and drug resistance of non-fermenting bacterial infection in intensive care unit (ICU) at a tertiary hospital during seven consecutive years,and to provide evidence for rational use of antibiotics in ICU.Methods A retrospective analysis was conducted.The related data about non-fermentative bacteria obtained from clinical specimens,collected from lower respiratory tract,blood,urine,bile and other secretions of ICU patients admitted to Binzhou Medical University Hospital from January 2009 to December 2015 were retrospectively analyzed.The distribution characteristics and drug resistance of non-fermentative bacteria,and isolation rate of multiple drug resistance (MDR) strains were analyzed.Results 2 672 strains of nonfermentative bacteria were isolated during seven consecutive years,accounting for 57.9％ gram negative (G-) bacilli (2 672/4 613),and 35.2％ of all bacteria (2 672/7 587).The top five were Acinetobacter baumannii (38.4％),Pseudomonas aeruginosa (34.6％),Onion burkholderia cepacia (9.9％),Stenotrophomonas maltophilia (6.2％),and Pseudomonas fluorescens (5.6％).Non-fermentative bacteria were mainly isolated from the lower respiratory tract (60.9％).Isolation of the non-fermentative bacteria accounted for over 50％ of G-bacilli during seven consecutive years,and the isolation rate of the top five types of bacteria showed no obvious change,while positive rate of Acinetobacter baumannii showed a tendency to increase (obviously from 26.5％ in 2009 to 50.2％ in 2015),and a lowering trend of positive rate of Onion burkholderia cepacia,Stenotrophomonas maltophilia,and Pseudomonas fluorescens was obvious (from 15.6％,10.6％,13.0％ in 2009 to 5.6％,7.4％,1.4％ in 2015 respectively) was observed.The isolation rate of Pseudomonas aeruginosa was stable (about 30％) during seven consecutive years.The drug susceptibility results showed that the resistant rates of Acinetobacter baumannii against imipenem,meropenem,aminoglycosides and third-generation cephalmsporins were all higher than 70％,while its resistant rate to cefoperazone-sulbactam was relatively lower (40.2％-68.1％)with relatively higher sensitivity rate (23.6％-46.0％).In contrast,the resistant rates of Pseudomonas aeruginosa against antibiotics were low,while the sensitivity rate to fourth-generation cephalmsporins cefepime (58.3％-87.7％)and third-generation cephalmsporins was high (ceftazidime:55.6％-79.3％,piperacillin-tazobactam:62.5％-86.2％,cefoperazone-sulbactam:46.0％-89.8％).From 2009 to 2015,the incidence of MDR strains of Acinetobacter baumannii showed an obvious increasing tendency (from 68.0％ to 84.1％);in contrast,the incidence of MDR strains of Pseudomonas aeruginosa did not show an obviously increase in incidence from 2009 to 2012,on the other hand,it showed a decreasing tendency from a peak 68.6％ in 2012 to 23.5％ in 2015.Conclusions The isolation rate of non-fermentative bacteria was high and the drug resistance situation was serious.Therefore,it is important to grasp the knowledge regarding distribution characteristics,drug resistance and variation of non-fermentative bacteria in ICU.It is not only beneficial for both rational use of antibiotics,improve efficacy but also helpful in reducing the emergence of drug resistance stains.

Research of 24-hour dynamic sleep monitoring and melatonin changes in patients with delirium in intensive care unit / 中华危重病急救医学

Objective To dynamic monitor and analyze the characteristic of polysomnography (PSG)and melatonin levels of delirium patients in intensive care unit (ICU). Methods A prospective observational study was performed from December 2013 to April 2014. The patients admitted to ICU of Affiliated Hospital of Binzhou Medical College for more than 72 hours were evaluated with confusion assessment method for the ICU (CAM-ICU),and were divided into delirium group and non-delirium group. Sleep patterns of all the patients underwent continuous PSG for up to 24 hours were evaluated. Melatonin levels were determined every 4 hours with enzyme linked immunosorbent assay (ELISA)duration sleep monitoring. Results Eighteen patients were enrolled,and 9 were delirium patients. All the patients had sleep disorders:a decrease in rapid eye movement (REM)sleep 〔(5.91±5.26)%〕,an increase in the sleep fragmentations 〔arousal index was (15.40 ±12.79)times/h〕,and the N3 sleep stage was on the lower limit of normal 〔(14.67 ±11.10)%〕. Compared with non-delirium group,the REM sleep was significantly decreased in delirium group〔(0.10±0.20)%vs.(8.83±3.81)%,t=4.782,P=0.001〕. Melatonin levels lost rhythm between day and night,and there was no difference in melatonin between delirium group and non-delirium group (time effect:F=1.370,P=0.287;between-group effect:F=1.646,P=0.250;interaction effect:F=1.558,P=0.247). The peak of melatonin levels of delirium group appeared on 06:00 〔(137.84±62.21)ng/L〕and 14:00 〔(148.24±58.8)ng/L〕, the minimum value on 22:00 〔(64.47 ±26.97) ng/L〕. But in non-delirium group,the peak of melatonin levels appeared on 02:00 〔(63.52 ±39.75)ng/L〕,the minimum value on 10:00 〔(44.87 ±11.19)ng/L〕. Conclusions ICU patients have sleep disorders,and the delirium patients have less REM stage. Normal rhythmic melatonin secretion changes of ICU patients were lost. The delirium peak of patients appears in the daytime.

The effect of apigenin on lipopolysaccharide-induced acute lung injury and its mechanism in mice / 中国中西医结合急救杂志

Objective To observe the effect of apigenin on acute lung injury (ALI) induced by lipopolysaccharide(LPS)in mice,and to discuss its possible mechanism. Methods Forty healthy male Kunming mice were randomly divided using random number table into control group,model group and low,medium,high dose groups of apigenin intervention,and each group consisted of 8 mice. The model of ALI was reproduced by intraperitoneal injection of 5 mg/kg LPS. Mice of the low,medium and high-dose intervention groups were given intraperitoneal injection of apigenin 10,25,50 mg/kg,respectively,1 hour before LPS modeling. Pathological changes in right upper lobe of lung tissue were examined after hematoxylin and eosin(HE)staining and pathology score was observed at 6 hours after modeling. Right inferior lung was weighed to measured wet/dry ratio(W/D). Intercellular adhesion molecule-1(ICAM-1)and tumor necrosis factor-α(TNF-α)in serum and bronchoalveolar lavage fluid (BALF)were determined by enzyme linked immunosorbent assay(ELISA). The mRNA expressions of p38 mitogen-activated protein kinase(p38MAPK),ICAM-1,and TNF-α were determined by reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with control group,lung W/D ratio in model group was significantly increased(17.79±2.89 vs. 5.56±0.37,P<0.05),and the pathology score was significantly elevated(10.32±0.23 vs. 1.87±0.54,P<0.05),ICAM-1 and TNF-α contents,in serum and BALF were increased〔ICAM-1(ng/L) in serum:21.4±2.7 vs. 14.3±3.5,TNF-α(ng/L)in serum:254.8±10.6 vs. 142.3±13.7;ICAM-1(ng/L)in BALF:20.3±2.4 vs. 11.5±3.2,TNF-α(ng/L)in BALF:230.3±5.8 vs. 110.5±11.2,all P<0.05〕,and the mRNA expressions of p38MAPK,ICAM-1 and TNF-α were also increased significantly(the mRNA expression of p38MAPK,ICAM-1 and TNF-αwere 4.42±0.37,4.89±0.27,3.28±0.13,respectively,all P<0.05). Different doses of apigenin could obviously alleviate the damaging effect to the lung,and the most obvious effect was seen in the medium dose group,in which lung W/D ratio was 13.28±1.21,ICAM-1 in serum was(18.5±4.3)ng/L,TNF-αin serum was(169.4±20.8)ng/L,ICAM-1 in BALF was(17.8±3.5)ng/L,TNF-αin BALF was(150.4±7.1)ng/L, the mRNA expression of p38MAPK,ICAM-1 and TNF-αin lung tissue was 2.99±0.28,3.97±0.17,2.87±0.27, respectively. Statistically significant difference was found when they were compared with that of model group(P<0.05 or P<0.01). Conclusion Different doses of apigenin have some antagonistic effect against LPS in producing ALI in mice,the best improvement effect was seen in the medium dose group,and the protective effect may be related to inhibition of p38MAPK signaling pathway activity and reduction of pro-inflammatory factors such as TNF-αand ICAM-1 expression.

Gene-transfected bone marrow mesenchymal stem cells combined with allogeneic bone for repair of sheep critical-size bone defects / 中国组织工程研究

BACKGROUND:Many in vivo and in vitro experiments indicate that implantation of exogenous basic fibroblast growth factor can significantly promote the process of bone formation, but the in vivo degradation of exogenous basic fibroblast growth factor affects the therapeutic efficacy. OBJECTIVE:To observe the effects of basic fibroblast growth factor-transfected mesenchymal stem cells which transfected using molecular biology techniques combined with al ogeneic bone in the repair of critical-size bone defects in sheep. METHODS:Al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells combined with al ogeneic bone material stents, al ograft bone material,β-tricalcium calcium material were respectively implanted into critical-size bone defects in sheep. After 4, 8 and 12 weeks, histological and immunohistochemical staining was performed. RESULTS AND CONCLUSION:At 12 weeks after implantation of al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells as tissue engineering bone, there were many cartilage-like structures in the operative binding region and a large amount of osteoblast-like cells in the center of operative region, and there was more material degradation in the entire operative area as compared with other groups;there were fibrous connective tissues ful of the pores, and osteoclast-like cells were commonly seen around the implant material;bone sialoprotein and col agen type Ⅰ expression were strongly positive. In the other three groups, although the cartilage-like structure appeared in the binding region, dead bone structure was found in the central area, and bone sialoprotein and type Ⅰ col agen expression was weak. These findings indicate that al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells can basical y repair critical-size bone defects in sheep.

Xenogeneic bone properties prepared by calcination and freeze-dried methods / 中国组织工程研究

BACKGROUND:Xenogeneic bone structure and biological characteristics are similar to human bone tissue, and the xenogeneic bone has a decreased antigenicity after physicochemical treatment, with a natural porous structure and rich source, and can be kept for a long time, which is considered to be an effective way to solve the shortage of the autogenous bone and al ograft bone. OBJECTIVE:To compare the physical and chemical properties of xenogenic bone materials prepared by two different methods. METHODS:Sheep cancellous bone treated with chemical method was placed into the muffle furnace at 1 000 ℃for 2 hours to prepare calcined bone. Another cancellous bone was placed into an 80 ℃ refrigerator for 4 weeks and then placed into a vacuum instrument to prepare freeze-dried bone. Cancellous bone rinsed with ultra-pure water served as controls. RESULTS AND CONCLUSION: Three groups of samples retained three-dimensional porous structure which similar with human bone tissue under microscopic observation. The framework was stil intact, with a smal pore of 55-650μm and high porosity of 65%-80%. For the calcined bone, the toughness was decreased and the brittleness increased significantly, but the freeze-dried bone had a little changes in the mechanical properties. Through diffraction analysis, hydroxyapatite was the main composition of the three groups. However, there was a smal amount ofβ-tricalcium phosphate in the calcined bone. Spectrum analysis confirmed that calcium and phosphorus content in these three groups were al close to the human body. The results suggest the cancellous bone treated with these two methods is similar to human bone structure, and the major elements are close to the body. In addition, the cancellous bone after processing has enough smal pore and higher porosity. However, calcination process has a more influence on the mechanical property of scaffold materials, and the freeze-dried bone has a little change but the antigen cannot be completely removed that can reach the basic requirements.

Clinical analysis of CT guided percutaneous radiofrequency thermocoagulation for treatment of trigeminal neuralgia / 中国基层医药

Objective To analyze and evaluate the clinical effect of CT guided percutaneous semilunar ganglion radiofrequency thermocoagulation for treatment of trigeminal neuralgia.Methods 52 patients with trigeminal neuralgia treated in our hospital were selected.They underwent the CT three-dimensional guidance on trigeminal neuralgia Ⅱ,Ⅲ,Ⅱ + Ⅲ neuralgia neuralgia patients with anterior foramen ovale puncture semilunar ganglion,verify the operative target,and a standard radiofrequency target temperature damage under general anesthesia.Results Of the 52 patients,the treatment effect was excellent in 49 cases,good in 1 patient,the effect of other 2 cases was poor,the excellent and good rate was 96.15％.Conclusion CT guided percutaneous puncture semilunar ganglion radiofrequency thermocoagulation in the treatment of patients with trigeminal neuralgia is simple,safe,and easy to be accepted by patients,the cure rate is high,it is worth learning and promotion.

Expression and significance of angiopoietin-2 and Tie-2 receptors in a rat model of acute lung injury / 中国病理生理杂志

AIM: To explore the expressions and significance of angiopoietin-2 (Ang-2) and tyrosine kinase with immunoglobulin and epidermal growth factor homology domains-2 (Tie-2) receptors in a rat model of acute lung injury (ALI). METHODS: Wistar rats (n=42) were divided into control group (n=12) and cecal ligation and puncture (CLP) group (n=30). Control group underwent sham operation, and CLP group underwent cecal ligation and puncture to make the model of ALI. 12 h after sham operation or CLP, 6 rats in each group were killed, and arterial blood gas analysis and lung coefficient were tested. The expressions of Ang-2 and Tie-2 receptors in lung tissue were observed by immunohistochemical method. Blood samples of the rest rats were collected from vena caudalis, and Ang-2 levels were measured by enzyme linked immunosorbent assay (ELISA). The mortality rate in each group within 36 h was compared. The lung architecture was observed under microscope. RESULTS: The lung architecture in control group was clear and intact. Alveolar septum was thicker, blood capillary was congested, and neutrophils and macrophages were infiltrated in the lung tissue in CLP group. Tie-2 receptors were expressed in bronchial epithelial cells, smooth muscle cells and endothelial cells in control group. Besides the similar expression as control group, high expression of Tie-2 on neutrophils and macrophages in CLP group was observed. In the adhesion location of Tie-2 receptors positive inflammatory cells, there was stronger staining in endothelial cells. Ang-2 was expressed in smooth muscle cells, bronchial epithelial cells and endothelial cells in control group. The Ang-2 level in CLP group were higher than that in control group [(8.14±1.74) μg/L vs (4.63±0.49) μg/L, P<0.01], and the Ang-2 level of dead rats was higher than that of survival rats within 36 h in CLP group [(8.95±1.61)μg/L vs (6.80±0.96)μg/L, P<0.01]. Oxygen partial pressure in control group was lower (P<0.01) and lung coefficient was higher (P<0.01) than that in CLP group. CONCLUSION: Ang-2 and Tie-2 receptors may participate in the pathophysiology of ALI, and Ang-2 level is correlated with mortality.

A non-invasive equation in diagnosis of idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia / 中华内科杂志

Objective Idiopathic pulmonary fibrosis (IPF) and nonspecific interstitial pneumonia (NSIP) are the two largest subsets of idiopathic interstitial pneumonia (IIP). They have a sharply differences in therapy and survival, however, the identification of them is difficult In general, NSIP has a much better responsiveness to therapy and good survival On the other hand, in IPF, was demonstrated to be associated with poorer survival and responsiveness to therapy. To diagnosis of IPF and NSIP need histopathologic classification which however, requires surgical lung biopsy. Therefore, we try to set up an equation using those data from non-invasive methods to identify IPF and NSIP. Methods A retrospective review of 32 patients with IPF (n = 14) and NSIP (n = 18) was carried out. General clinical data, pulmonary function, chest radiographic, and bronchoalveolar lavage fluid were recorded. The statistical methods of logistic regression and multiple linear regression are used to establish the equation. The method of curve fitting is used to find the critical value of the equation to the differential diagnosis between the IPF and NSIE Results (1) Compare to NSIP, patients of IPF are older, more males than females, having a bigger proportion of smokers. (2) Compared with NSIP patients, IPF patients have higher CT score of "grid shadow", "cellular shadow" and lower CT score of "ground-glass shadow". (3) Bronchoalveolar lavage (BAL) lymphocytosis was more frequently observed in NSIP than IPF. (4) We get an equation: y = 0. 9 + 0. 123x1 -0. 045x2 +0. 009x3 +0. 033x4 and it can improve the differential diagnosis between the IPF and NSIP in this group of patients. Conclusion Clinical,high resolution computerized tomography and BAL are useful non-invasive tools in diagnosis IPF and NSIP. The equation: y = 0.9 + 0. 123x1 - 0.045x2 + 0. 009x3 +0. 033x4 can help us to distinguish the IPF and NSIP.

Doppler studies of maternal uterine artery and fetal circulation in women with early-onset of severe preeclampsia / 中华围产医学杂志

Objective To observe the hemodynamic variations of both the mother and fetus when the pregnancy was complicated with early-onset of severe preeclampsia, and to evaluate the role of monitoring the blood flow with Doppler in the management and prognostic value in the same condition. Methods A prospective case-control study was conducted in 36 pregnant women admitted for early-onset of severe preeclampsia (S-PE). The control group included 72 healthy pregnant women matched with the S-PE group. All had color Doppler sonography monitored during this study period. Data were collected for the umbilical artery (UA) pulsatility index (PI) and systolic/diastolic ratio (S/D) , the middle cerebral artery (MCA) PI and S/D and the maternal mean uterine artery (UtA) PI and S/D. Those subjects with adverse perinatal outcomes were followed up after birth. Results The mean Doppler indices(S/D and PI) of fetal UA, MCA and maternal UtA were all declined gradually with the advancing gestational ages in both groups. The values of UA-S/D, UA-PI, UtA-S/D, and UtA-PI in the early-onset S-PE group were significantly higher than those of the control(P