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Objective:To study the signaling pathway of the up-regulation of claudin-5 expression by Xuebijing injection.Methods:Animal and cell models of acute respiratory distress syndrome (ARDS) were induced by lipopolysaccharide (LPS). ① In vivo study, 20 male Sprague-Dawley (SD) rats were randomly divided into 4 groups: control group, LPS group (LPS injection 10 mg/kg for 12 hours), Xuebijing control group (Xuebijing injection 1 mg/kg, twice a day, for 3 days), and Xuebijing intervention group (LPS injection after pretreatment of Xuebijing injection), according to random number method with 5 rats in each group. The lung tissues were taken to detect lung dry/wet weight ratio (W/D) and the morphological changes in each group. Claudin-5, phosphorylated forkhead box transcription factor O1 (p-FOXO1), total FOXO1 (t-FOXO1), phosphorylated Akt (p-Akt) and total Akt (t-Akt) in lung tissues were detected by immunohistochemical staining (IHC) and Western blotting. ② In vitro study, human pulmonary microvascular endothelial cells (HPMECs) were divided into 6 groups (5 holes in each group): control group, Xubijing control group (incubated with 2 g/L Xubijing for 24 hours), phosphoinositide 3-kinases (PI3K) signaling pathway LY294002 control group (incubated with 10 μmol/L LY294002 for 1 hour), LPS group (incubated with 1 mg/L LPS for 12 hours), Xubijing intervention group (incubated with 2 g/L Xuebijing for 24 hours, then with 1 mg/L LPS for 12 hours) and LY294002 intervention group (incubated with 10 μmol/L LY294002 for 1 hour, then with 2 g/L and Xubijing for 24 hours, and then with 1 mg/L LPS for 12 hours). The expression levels of claudin-5, p-FOXO1, t-FOXO1, p-Akt and t-Akt of HPMECs in each group were assessed by Western blotting. Results:In vivo study: ① Compared with the control group, the lung W/D ratio increased significantly in LPS group (6.79±0.42 vs. 4.19±0.13), and decreased significantly after the intervention of Xuebijing (4.92±0.38 vs. 6.79±0.42, P < 0.01). ② Morphological changes of lung tissue: compared with the control group, the injury of lung tissue in LPS group was more serious, which was significantly improved after Xuebijing intervention. ③ Expression levels of claudin-5, p-Akt/t-Akt and p-FOXO1/t-FOXO1: the expression levels of claudin-5, p-Akt/t-Akt and p-FOXO1/t-FOXO1 in LPS group were significantly decreased as compared with the control group (claudin-5/GAPDH: 0.33±0.03 vs. 1.03±0.07, p-Akt/t-Akt: 0.18±0.02 vs. 1.01±0.13, p-FOXO1/t-FOXO1: 0.16±0.06 vs. 1.00±0.19, all P < 0.01). After the intervention of Xuebijing, the expression levels were significantly increased as compared with the LPS group (claudin-5/GAPDH: 0.53±0.05 vs. 0.33±0.03, p-Akt/t-Akt: 0.56±0.12 vs. 0.18±0.02, p-FOXO1/t-FOXO1: 0.68±0.10 vs. 0.16±0.06, all P < 0.01). In vitro study: compared with the control group, the expression level of claudin-5 in the LPS group was significantly decreased (claudin-5/β-actin: 0.45±0.03 vs. 1.01±0.15, P < 0.01), and the expression level of claudin-5 in Xuebijing intervention group was also significantly decreased (claudin-5/β-actin: 0.80±0.08 vs. 1.01±0.15, P < 0.01). After the intervention of LY294002, the expression of claudin-5 was significantly decreased as compared with the Xubijing intervention group (claudin-5/β-actin: 0.41±0.02 vs. 0.80±0.08, P < 0.01). Conclusion:Xuebijing injection improve pulmonary vascular barrier function in rats with ARDS by up-regulating claudin-5 expression through PI3K/Akt/FOXO1 signaling pathway.
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Breast cancer in younger patients exhibits stronger invasiveness and poorer outcomes compared with that in elders. Cut-off for young age remains unclear, and its basis as prognostic factor representing a continuous variable requires further investigation. Preg-nancy-associated breast cancer in young patients also possesses unique biological features. In this review, we define breast cancer di-agnosed in patients below 25 years old as very young breast cancer and summarize recent studies based on genetic factors, clinicopa-thology, prognostic characteristics, and pregnancy-associated breast cancer to assist in management of very young breast cancer pa-tients.
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ObjectiveTo investigate the early clinical risk factors of severe acute inhaled organic fluorine poisoning.Methods The clinical data of patients with acute poisoning of organic fluorine inhalation admitted since 2004 in Northern Jiangsu People's Hospital were retrospectively analyzed. According toDiagnostic Criteria of Occupational Acute Fluorohydrocarbon Poisoning(GBZ66-2002), all the patients were divided into three groups: mild, moderate and severe poisoning groups, the severe cases were included in the intensive group, and the others were grouped in the non-intensive group. The contents in the survey were as follows: gender, age, vital signs on admission (body temperature, pulse rate, respiratory rate, systolic blood pressure), arterial blood gas analysis record〔arterial oxygen saturation(SaO2), oxygenation index(PaO2/FiO2), lactic acid(Lac) and arterial partial pressure of carbon dioxide(PaCO2), pH value(pH)〕. Before treatment, the white blood cell(WBC) count, platelet(PLT) count, levels of alanine transaminase(ALT), creatinine(Cr), blood glucose, electrolytes(potassium, sodium, chloride, calcium), creatine kinase isoenzyme(CK-MB), etc. were examined and recorded. All the patients were immediately arranged for bedside chest X-ray examination, and the chest X-ray lung injury scores were recorded. By univariate and multivariate logistic regression analyses, the receiver operating characteristic curve(ROC curve) was drawn to evaluate the diagnostic value of the clinical risk factors.Results Sixty-two cases consisting with the standard criteria of enrollment were collected in the study, 36 cases being in intensive group and 26 cases in non-intensive group. The univariate analysis showed that the differences in pulse rate, respiratory rate, PaO2/FiO2, WBC, SaO2, Lac, pH, and lung injury score were statistically significant(P<0.05 orP<0.01). Logistic multiple regression analysis showed that PaO2/FiO2, WBC, Lac and chest X-ray lung injury score were the four indexes for predicting the independent risk factors of severe acute inhaled organic fluorine poisoning. The area under ROC curve(AUC) of PaO2/FiO2 was the highest(0.884), 95% confidence interval(95%CI) was 0.784 - 0.984, the critical value was 96.5 mmHg(1 mmHg=0.133 kPa), with the sensitivity of 75.6%, specificity of 95.2%, positive predictive value(PPV) of 92.3% and the negative predictive value(NPV) of 71.4%, in sequence, the rest were WBC(AUC 0.846, 95%CI 0.728 - 0.965, the criticalvalue 12.15×109/L), Lac(AUC 0.800, 95%CI 0.662 - 0.938, the critical value 4.2 mmol/L), chest X-ray lung injury score(AUC 0.795, 95%CI 0.652 - 0.938, the critical value 2.50), the sensitivity of the above three items was 90.2%, 83.6%, 88.5%, specificity was 90.2%, 83.6%, 88.5%, the PPV was 86.7%, 82.4%, 85.8% and NPV was 72.0%, 73.9%, 69.2% respectively.ConclusionThe blood WBC count, Lac, PaO2/FiO2 and chest X-ray lung injury score can be used as the early clinical risk factors of severe acute inhaled organic fluorine poisoning.
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Objective To investigate the expression and clinical significance of platelet activating factor [PAC]-1, CD62P and TPP hi severe sepsis. Method Patients with severe sepsis who were admitted into the EICU of Subei People's Hospital from April 2007 to March 2008 were included. Patients with severe sepsis (Group Ⅲ)were treated according to the treatment guidelines for severe sepsis, and were divided, according to their clinical records, into those who survived and those who died within 28 days of admission. Patients admitted during the same period with symptoms of infection but without severe sepsis were included as the General Infected Group (Group Ⅱ). A Control Group (Group Ⅰ) comprised patients who visited the hospital over the same period for physical examination or the healthy volunteers. The group members were all included randomly, and the gender and sex of patients in all three groups were similar. Patients with acute brain infarction, acute coronary syndrome,serious diabetes, hyperlipidemia, malignant tumor, leukemia, primary liver, renal and hematopoietic system dis-eases,long-term bedridden patients, pregnant women, and patients taking hormone treatment or hranunosuppres-sants were excluded from the study. Morning venous blood was collected and ELISA and Flow Cytometry performed on the fwst day of admission for Groups Ⅰ- and Ⅱ, and on the first, third and fifth day after admission for Group Ⅲ, to determine the TpP,PAC-1 and CD62P respectively; and the Marshall score was determined. Data were ana-lyzed by SPSS 12.0 software. For continuous variables, comparisons among groups were analyzed by ANOVA.Levene's and LSD test were applied to assess homogeneity. Bivariate test is applied to Correlation Analysis. P<0.05 was regarded as a statistically significant difference. Results There were a total of 20 patients each in GroupⅠ-and GroupⅡ, and 30 in Group Ⅲ; of these, 19 were classed as survivors and 11 died during the 28-day peri-od. On the first day of admission, there were no significant differences in PAC-1, CD62P or TpP expression between Groups Ⅰ- and Ⅱ(P>0.05); however, Group Ⅲ was significantly different compared with both Group Ⅰ and Group Ⅱ (both:P<0.05). The expression of PAC-1, CD62P and TpP tended to decline in the survivor group,and became normal with the treatment process, while the expression of PAC-1 ,CD62P and TpP in the patients who died remained high, and even increased significantly over time. On the first day, the expression of CD62P and TpP in the patients who survived and in those who died was not significantly different (P>0.05); on the third day,however, a significant difference appeared with values of (2.89±1.48) % vs. (5.04±2.57) % (P<0.01) for CD62P, and (5.24±2.22) mg/L vs. (9.20±1.93) mg/L (P<0.01) for TpP. The expression of PAC-1 was significantly different between the two subgroups on the first day, with values of (3.15±0.42)% vs. (5.30±.48)% (P<0.01). The Marshall score of the two groups showed similar changes. Correlation analysis showed that PAC-1, CD62P and TpP were significantly correlated with the Marshall score. Conclusions Platelet activation and microthrombosis existing in the early stage of severe sepsis work together in the early hypercoagulable state.They both play important roles in disease development and progression. The dynamic detection of CD62P and TpP is beneficial to the diagnosis and prognosis of severe sepsis.PAC-1 appears to hold a risk stratification effect, as pa-tients with high expression of PAC-1 in the early stage show poor prognosis. Therefore, PAC-1 could be used as a marker of severe sepsis and poor prognsis.