Value of vascular endothelial-cadherin in evaluating the severity of patients with sepsis / 中华危重病急救医学

Objective:To explore the value of vascular endothelial-cadherin (VE-cad) in evaluating the severity of sepsis.Methods:A prospective study was conducted to select 85 patients with sepsis treated in the emergency ward of the First Affiliated Hospital of Wenzhou Medical University from June 1, 2015 to November 1, 2017. The gender, age, medical history, first infection site, number of affected organs, laboratory indexes, acute physiology and chronic health evaluationⅡ(APACHEⅡ), simplified acute physiology score Ⅱ(SAPSⅡ), sequential organ failure assessment (SOFA) and the total length of stay, emergency intensive care unit (EICU) length of stay, 28-day at admission and survival during hospitalization were measured, and the VE-cad level within 24 hours at admission was measured. The patients were divided into sepsis group and septic shock group according to the progress of the disease. The patients were divided into multiple organ dysfunction syndrome (MODS) group and non MODS group according to whether they were accompanied by MODS. The differences of the above indexes in patients with different disease progression, MODS and different prognosis were analyzed and compared. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the value of VE-cad in evaluating the severity of sepsis.Results:A total of 85 patients were included, mainly respiratory tract infection. Among them, 38 cases were sepsis and 47 cases were septic shock, 39 cases had MODS, 46 cases had no MODS, 64 cases survived and 21 cases died within 28 days after admission. Compared with sepsis group, the number of affected organs in septic shock group was greater [3 (2, 4) vs. 1 (0, 2)], APACHE Ⅱscore [13 (10, 21) vs. 7 (5, 12)], SAPS Ⅱscore [35 (31, 55) vs. 7 (5, 12)], SOFA score [7.0 (5.0, 10.0) vs. 3.0 (0, 5.0)], blood lactic acid [Lac (mmol/L): 3.5 (2.4, 6.2) vs. 1.9 (1.2, 2.2)], C-reactive protein [CRP (mg/L): 90.0 (58.1, 90.0) vs. 50.5 (38.0, 90.0)] and VE-cad levels [mg/L: 1.427 (1.141, 2.150) vs. 1.195 (0.901, 1.688)] were significantly increased, while platelet count [PLT (×10 9/L): 113.4±67.2 vs. 202.5±109.5] and hemoglobin (Hb) levels (g/L: 106.3±36.3 vs. 118.6±18.0) were significantly decreased (all P < 0.05). Compared with non MODS group, APACHE Ⅱ score [14 (10, 22) vs. 8 (6, 13)], SAPS Ⅱ score [36 (32, 56) vs. 29 (24, 35)], SOFA score (7.9±3.9 vs. 4.0±3.8), in-hospital mortality [53.8% (21/39) vs. 0% (0/46)], Lac [mmol/L: 3.1 (2.3, 6.3) vs. 2.1 (1.4, 4.6)] and VE-cad levels [mg/L: 1.427 (1.156, 1.937) vs. 1.195 (0.897, 1.776)] in MODS group were significantly higher, the length of stay in EICU was significantly longer [days: 6 (3, 12) vs. 3 (0, 7)], and the PLT level was significantly lower (×10 9/L: 118.2±80.0 vs. 182.5±104.0, all P < 0.05). Compared with the death group, the number of affected organs in the survival group was fewer [2 (1, 3) vs. 3 (1, 5)], APACHE Ⅱ score [9 (6, 13) vs. 21 (13, 25)], SAPS Ⅱ score [31 (25, 36) vs. 55 (35, 63)] and SOFA score (4.7±3.7 vs. 8.9±4.5) were significantly reduced, and the length of stay in EICU [days: 4 (1, 8) vs. 8 (3, 15)] was significantly shorter (all P < 0.05). ROC curve analysis showed that area under the ROC curve (AUC) of VE-cad, SOFA score and VE-cad combined with SOFA score in evaluating the severity of sepsis were 0.632 [95% confidence interval (95% CI) was 0.513-0.750], 0.830 (95% CI was 0.744-0.916) and 0.856 (95% CI was 0.779-0.933), respectively. When the cut-off value of VE-cad was 1.240 mg/L, the sensitivity was 68.1% and the specificity was 55.3%, the sensitivity of VE-cad combined with SOFA score was 85.1%, the specificity was 73.7%. Conclusion:VE-cad has a certain evaluation value for the severity of sepsis, and the evaluation value of VE-cad combined with SOFA score is better than that of VE-cad single index.

Predictive value of the C-reactive protein/albumin ratio in tsutsugamushi disease complicated with organ function damage / 中华急诊医学杂志

Objective:To explore the predictive value of the serum C-reactive protein (CRP)/albumin (ALB) ratio (CAR) for organ damage in tsutsugamushi disease.Methods:The clinical data of 166 patients with tsutsugamushi disease admitted to the First Affiliated Hospital of Wenzhou Medical University from January 1, 2010 to December 31, 2020 were retrospectively analyzed. The patients were divided into the organ damage group (72 cases) and non-organ damage group (94 cases) according to the organ damage criteria. The general data and laboratory test results of the two groups of patients were compared. The significant indicators of univariate analysis were analyzed by multivariate logistic regression analysis. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to analyze the predictive value of CAR for organ damage in patients with tsutsugamushi disease.Results:There were no significant differences in age, sex, days of fever, and admission body temperature between the organ damage group and non-organ damage group ( P>0.05). However, the body mass index, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), length of hospital stay, hospitalization expense, percentage of neutrophils (NEUT), lymphocyte count, procalcitonin, CRP, and CAR in the organ damage group were significantly higher than those in the non-organ damage group ( P<0.05), and ALB was significantly lower than that in the non-organ damage group ( P<0.05). Multivariate logistic regression analysis showed that APACHEⅡ( P=0.039), NEUT ( P=0.003), and CAR ( P=0.011) were independent risk factors for tsutsugamushi disease complicated by organ damage. The ROC curve showed that the AUCs of APACHEⅡ, NEUT, and CAR were 0.655, 0.716, and 0.727, respectively. When the cut-off value of CAR was 2.86, the sensitivity was 55.6%, and the specificity was 79.8%. Conclusions:Elevated CAR is an independent risk factor for tsutsugamushi disease complicated with organ damage and can be used as an important indicator to evaluate the presence or absence of organ damage in patients with tsutsugamushi disease.

Risk factors analysis and early warning evaluation model construction of disease severity in patients with dangerous upper gastrointestinal bleeding / 中华急诊医学杂志

Objective:To explore the influencing factors of severity of upper gastrointestinal bleeding (UGIB) and to establish the early warning evaluation model in the form of line chart, so as to provide a feasible basis for emergency nurses' triage.Methods:A total of 680 UGIB patients admitted to the Emergency Department of the First Affiliated Hospital of Wenzhou Medical University from January 2019 to January 2020 were retrospectively analyzed. They were divided into a modeling group ( n=510) and a validation group ( n=170) by random number table method, and were divided into a high-risk group and a low-risk group according to the expert Consensus on Emergency Diagnosis and Treatment Procedures for Acute Upper Gastrointestinal Bleeding in 2020. The differences of various indicators between groups were compared, the factors affecting the severity of the disease were analyzed by Logistic regression, and the nomogram was drawn and validated. Results:Multivariate logistic regression analysis showed that hematemesis ( OR=3.875, 95% CI: 2.212-6.79), diabetes ( OR=2.64, 95% CI: 1.184-5.883), syncope ( OR=10.57, 95% CI: 3.675-30.403), heart rate ( OR=3.262, 95% CI: 1.753-6.068), red blood cell distribution width ( OR=3.904, 95% CI: 2.176-7.007), prothrombin time ( OR=3.665, 95% CI: 1.625-8.269), lactic acid ( OR=3.498, 95% CI: 1.926-6.354) and hemoglobin ( OR=4.984, 95% CI: 2.78-8.938) were the influencing factors of the severity of UGIB patients ( P < 0.05). The nomogram model showed good consistency and differentiation (C-index=0.903, 95% CI: 0.875-0.931), and was verified internally (C-index=0.895) and Hosmer-Lemeshow goodness-of-fit test ( P=0.7936). Externally verified C-index was 0.899 (95% CI: 0.846-0.952). The calibration curve prompt warning evaluation model had good stability and the prediction efficiency was better than the modified early warning score ( P < 0.05). Conclusions:The early warning evaluation model has a reliable predictive value, which can provide a reference for emergency medical staff to screen high-risk patients and formulate targeted nursing interventions.

Analysis of prognostic factors in patients with Vibrio vulnificus sepsis / 中华急诊医学杂志

Objective:To explore the prognostic factors of patients with Vibrio vulnificus sepsis. Methods:The clinical data of 67 patients with Vibrio vulnificus sepsis from January 2008 to December 2019 in the First Affiliated Hospital of Wenzhou Medical University were retrospectively analyzed. Univariate analysis was used to compare the differences in general information, clinical manifestations, admission laboratory indicators, antibiotics and surgery between the death group and the cured group. Then the factors with significant difference in univariate analysis were included in multivariate analysis, and the factors of prognosis were obtained. Results:Univariate analysis showed that there were significant difference in liver disease, admission with hypotension shock, multiple limb injuries; admission leukocytes, platelets, pH value, albumin, lactic acid, aspartate aminotransferase, creatinine, procalcitonin, creatine kinase, activated partial thromboplastin time, prothrombin time between the death group and the cured group (all P <0.05). Multivariate analysis showed that admission lactate ( OR=0.628, 95% CI: 0.461-0.855, P=0.003), albumin ( OR=1.330, 95% CI:1.062-1.667, P=0.013), creatine kinase ( OR=0.999, 95% CI: 0.998-1.000, P=0.016) and admission surgery time ( OR=0.118, 95% CI: 0.015-0.938, P=0.043) were risk factors of the prognosis. Patients with high lactate, creatine kinase and low albumin at admission indicate poor prognosis; patients with admission surgery time≤ 12 h have better prognosis. Conclusion:For the treatment of patients with Vibrio vulnificus sepsis, medical staff should dynamically evaluate these prognostic factors in the early stage, and early surgical treatment should be adopted to improve the prognosis of patients.

Curcumin upregulates mitofusin 2 to alleviate acute lung injury in septic mice / 中华急诊医学杂志

Objective To investigate the protective effect and mechanism of curcumin on acute lung injury in septic mice.Methods Totally 120 clean BALB/c male mice were randomly (random number) divided into 8 groups:sham group,sepsis group,curcumin control group,curcumin intervention group,negative virus-sepsis group,negative virus-curcumin intervention group,Mfn2 interference-sepsis group,and Mfn2 interference-curcumin intervention group,15 rats in each group.Mice in the sepsis and the curcumin groups were given the cecal ligation and puncture (CLP) mice in the curcumin intervention and curcumin control groups were given curcumin 200 mg/(kg·d) for 1 week,and mice in the negative virus-sepsis group and negative virus-curcumin intervention groups were established by injection of a negative adeno-associated virus in the tail vein.The Mfn2 interference-sepsis and Mfn2 interference-curcumin intervention groups were established by injecting an adeno-associated virus carrying the Mfn2 interference sequence through the tail vein.Mice were sacrificed after 24 h in each group.The degree of lung injury was examined by lung wet-to-dry weight ratio and pathological examination.The inflammatory factors of alveolar lavage fluid including TNF-α and IL-6 were detected by ELISA,the activation of caspase-3,a key molecule for apoptosis,was detected by Western blot,and apoptosis was detected by TUNEL.The data were analyzed by SPSS 22.0 software,the count data was analyzed by x2 test,and the comparison of measurement data between groups was analyzed by one-way ANOVA.Results Compared with the sham group,the wet-to-dry weight ratio of lung tissue in the sepsis group was significantly increased (71.11 ± 3.78 vs 31.11 ± 5.61,P=0.002),the histopathological score was significantly higher (P=0.006),the inflammatory factors TNF-α (P=0.001) and IL-6 (P=0.012) were dramatically increased,and the apoptosis of lung tissue and the expression of caspase-3 cleaved were also significantly increased (P=0.001).Compared with the sepsis group,the wet-to-dry weight ratio and the histopathological score of lung tissue in the curcumin-treated group was significantly lower (32.84 ± 6.15 vs 71.11 ± 3.78,P=0.004),and the inflammatory factors TNF-α(P=0.013) and IL-6 (P=0.003) were obviously decreased,and apoptosis and apoptosis-related protein caspase-3 cleaved expression were also dramatically decreased (P=0.012).After Mfn2 was down-regulated,Mfn2 interference-curcumin intervention group interfered with Mfn2.Compared with the sepsis group,the dry-to-wet weight ratio and the histopathological score of the lung tissue of the mice was not significantly decreased.Further studies found that after down-regulating Mfn2,compared with the Mfn2 interfere-sepsis group,Mfn2 interfere-curcumin intervention group had no such performance.The inflammatory factors TNF-α and IL-6 were not significantly decreased,and the apoptosis of lung tissue and the expression of apoptosis-related protein caspase-3 cleaved were not significantly reduced.Conclusion Curcumin may attenuate acute lung injury in sepsis by up-regulating the expression of Mfn2.

Antioxidant mechanism of HDAC2 regulating Nrf2 acetylation in LPS-induced type Ⅱ alveolar epithelial cell injury / 中华急诊医学杂志

Objective To explore the antioxidant mechanism ofhistone deacetylase 2 (HDAC2) regulating Nrf 2 acetylation in lipopolysaccharide (LPS)-induced type Ⅱ alveolar epithelial cell injury.Methods The experiment was divided into two parts.The first part was the routine culture of type Ⅱ alveolar epithelial cells of mice.The cells were stimulated with different concentrations of LPS (10 ng/ mL,100 ng/mL and 1 000 ng/mL).CCK-8 was used to detect the cell activity at 0 h,6 h,12 h,24 h and 48 h,respectively.The second part:Alveolar epithelial cells of type Ⅱ were cultured and divided into the normal control group (control group),LPS group,HDAC2 lentivirus interference group (siRNA-HDAC2 group) and HDAC2 lentivirus overexpression group (LV-HDAC2 group).The expression of HDAC2 and Nrf2 were detected by Western blot,the acetylation of Nrf2 was detected by immunoprecipitation,and the stability of nrf2 was detected after actinidone action.The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by chemical colorimetry.SPSS 23.0 statistical software was used.LSD-t test was used for comparison between two groups,and one-way ANOVA test was used for comparison among multiple groups.Results Compared with the control group,the expression of HDAC2 protein in the LPS group increased (t=5.974,P=0.027),the acetylation level of Nrf2 decreased (t=7.223,P=0.002),the Nrf2 protein level increased (t=2.929,P=0.043),the protein stability of Nrf2 increased,the SOD activity decreased (t=121,P＜0.01),and the MDA content increased (t=10.45,P=0.000 5).Compared with the LPS group,Nrf2 acetylation level decreased in the LV-HDAC2 group (t=1 1.29,P=0.000 4),Nrf2 protein expression increased (t=3.194,P=0.033),Nrf2 protein stability increased,SOD activity increased (t=4.678,P=0.009),and MDA content decreased in the LV-HDAC2 group (t=5.417,P=0.005 6).While the opposite trend was observed in the siRNA-HDAC2 group.Conclusion After LPS stimulation,oxidative stress of type Ⅱ alveolar epithelial cells was aggravated.HDAC2 could decrease the level of Nrf2 acetylation,increase the expression of Nrf2 protein,and alleviate LPS-induced oxidative stress.

Effect of Xuebijing on paraquat-induced cell oxidative stress and inflammation level in human renal proximal convoluted tubular epithelial cell-2 / 中国中西医结合急救杂志

Objective To investigate the effect of Xuebijing on paraquat-induced oxidative stress and inflammation level in human kidney cell line-2 (HK-2) cells. Methods The HK-2 cells were routinely cultured and divided into blank control group, paraquat poisoning model group, Xuebijing injection pretreatment group and Xuebijing control group according to random number table method. The concentration of paraquat in HK-2 cells were measured by high performance liquid chromatography (HPLC); the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) were measured by chemical colorimetry method; the levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA). Results With prolonged treatment, paraquat concentration gradually increased in HK-2 cells in the paraquat poisoning model group and the Xuebijing injection pretreatment group, reaching the peak value after treatment for 48 hours, moreover, the concentration of paraquat in HK-2 in the Xuebijing injection pretreatment group was significantly lower than that in the paraquat poisoning model group (mg/L: 4.26±0.20 vs. 5.77±0.18, P < 0.05). Compared with the blank control group, the contents of MDA, TNF-α and IL-6 of paraquat poisoning model group were obviously elevated [MDA (nmol/mg): 4.47±0.10 vs. 2.21±0.08, TNF-α (ng/L): 206.91±13.22 vs. 98.14±5.67, IL-6 (ng/L): 253.33±5.22 vs. 116.97±13.54, all P <0.05], and the activity of SOD was significantly reduced (U/mg: 33.30±0.62 vs. 41.58±0.17, P < 0.05). Compared with paraquat poisoning model group, the contents of MDA, TNF-α and IL-6 in Xuebijing injection pretreatment group were obviously decreased [MDA (nmol/mg): 2.92±0.17 vs. 4.47±0.10, TNF-α (ng/L): 166.29±15.47 vs. 206.91±13.22, IL-6 (ng/L): 209.39±3.18 vs. 253.33±5.22, all P < 0.05], and the activity of SOD was significantly increased (U/mg:38.10±0.67 vs. 33.30±0.62, P < 0.05) in the paraquat pretreatment group. Conclusion Xuebijing could reduce the concentration of paraquat in HK-2 cells, and decrease oxidative stress and inflammation factor levels in HK-2 cells.

Protective effect of Baicalin on lipopolysaccharide induced H9C2 cell inflammation and its mechanism / 中华急诊医学杂志

Objective To investigate the protective effect of Baicalin on inflammation induced by lipopolysaccharide in H9C2 cardiomyocytes and its possible mechanism. Methods H9C2 myocardial cells were cultured and pretreated with baicalin at the final concentration of 10, 20, 30 μmol/L for 12 hours, then stimulated with LPS at the final concentration of 1 μg/mL for 6 hours. The control group was treated with the same amount of saline to collect cell samples. CCK-8 (The Cell Counting Kit-8) was used to detect cell activity, enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of interleukin-6 (IL-6), interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α), Western blot was used to detect the protein expression levels of NF-κB p65, p-NF-κB p65, p38 MAPK, p-p38 MAPK, IκBα and p-IκBα. SPSS 23.0 statistical software was used. Independent sample t test was used for comparison between two groups, and one-way ANOVA test was used for comparison among multiple groups. Results The survival rate of myocardial cells in the control group was (93.67 +1.453)％. Compared with the control group, the survival rate of H9C2 myocardial cells induced by LPS decreased (P< 0.05). In the control group, the expression of IL-6 in H9C2 myocardial cells was (49.33 +2.42) pg/mL, the expression of TNF-α was (86.33 +1.85) pg/mL, and the expression of IL-1β was (28.67 +4.66) pg/mL. Compared with the control group, the expression levels of IL-6, IL-1β and TNF-α in H9C2 myocardial cells increased after LPS induction (P< 0.05), while the levels of p-NF-κ B p65, p-p38 MAPK and p-I κ B α protein increased (P< 0.05), while the levels of I κ B α protein decreased (P< 0.05), while the expressions of NF-κ B p65 and p38 MAPK protein did not change significantly (P> 0.05). Compared with LPS group, the survival rate of H9C2 myocardial cells in baicalin intervention group increased (P<0.05), the expression levels of IL-6, IL-1β and TNF-a decreased (P < 0.05), the levels of p-NF-κB p65, p-p38 MAPK, p-I κBα protein decreased (P< 0.05), and the level of IκBα protein increased (P< 0.05), while the expression of NF-κB p65 and p38 MAPK did not change significantly. (P>0.05). Conclusions Baicalin may alleviate LPS-induced cardiomyocyte inflammation by inhibiting the activation of NF-kappa B and p38 MAPK, and improve cell survival.

The protective effect and mechanism of somatostatin on the lung injury of paraquat intoxicated mice / 中华劳动卫生职业病杂志

Objective@#To explore the protective effect and mechanism of Somatostatin (SS) on the mice with Paraquat (PQ) poisoning, and to provide theoretical basis for clinical treatment of PQ poisoning.@*Methods@#48 SPF male BALB/c mice were randomly divided into control group, SS group, PQ group and PQ+SS group, with 12 mice in each group. 20 ml/kg SS solution was intraperitoneally injected into the SS group and PQ+SS group, and the same amount of normal saline was intraperitoneally injected into the PQ group and control group. After 1 hour of the above treatment, PQ group and PQ+SS group were given 60 mg/kg PQ solution by one-time gavage, while the control group and SS group were given the same amount of normal saline by gavage. After the above treatment for 3 hours, the SS group and PQ+SS group were intraperitoneally injected with SS solution (20 ml/kg) again, and the PQ group and the control group were intraperitoneally injected with the same amount of normal saline. 6 eyeballs were randomly selected from each group for blood collection, and the levels of TNF-α, MPO and il-6 in the blood of mice were detected by ELISA and other methods. The left lung was taken after blood collection to calculate the D/W ratio. The levels of SOD, caspase-3 and MDA were detected in some lung tissues by chemical colorimetry, and the amount of NF-κB was detected by Western blot. The lung histopathological changes were observed under light microscope.@*Results@#The mice in the control group and SS group showed normal activity and good general condition; Mice in the PQ group ate less and moved less, responded slowly to stimulation, breathed shallow and fast with thickened breath sound, had messy and dull fur, and had varying degrees of cyanosis on their lips and limbs; The above performance of PQ+SS group was less than that of PQ group. Under the light microscope, the alveolar structure of PQ group was disordered and seriously damaged. The pathological changes of lung tissue in PQ+SS group were significantly improved compared with that in PQ group, and the pathological scores were decreased (all P<0.05) . Compared with the control group, there was no significant change in the levels of various indicators in the SS group (all P>0.05) . While the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 in PQ mice were significantly increased, and the levels of oxidative cytokines SOD and lung tissue D/W were significantly decreased (all P<0.05) . Compared with the PQ group, the PQ+SS group, the levels of inflammatory cytokines (IL-6, TNF-α, Protein expression of NF-κB) , oxidative cytokines (MPO, MDA) and apoptotic cytokines caspase-3 decreased, and the levels of oxidative cytokines SOD and lung tissue D/W increased (all P<0.05) .@*Conclusion@#PQ poisoning can cause lung injury in mice, while SS can alleviate lung injury in PQ poisoned mice, improve the general survival state of mice, and play a certain protective role in lung injury caused by PQ poisoning, which may be achieved by SS throμgh reducing the inflammatory response, oxidative stress response and lung tissue apoptosis in lung injury caused by PQ poisoning.

Clinical analysis of 22 cases of syndrome caused by acute poisoning / 中华劳动卫生职业病杂志

Objective@#To analyze the clinical characteristics, treatment and prognosis of rhabdomyolysis (RM) caused by acute poisoning.Summarize the clinical characteristics and treatment experience, pay attention to the complications and improve the quality of rescue.@*Methods@#We collecte and summarize the clinical data, treatment and prognosis of 22 cases of RM caused by acute poisoning.@*Results@#We found that 21 patients (95.5%) had muscle damage, 13(59.1%) with coma, 8(36.4%) with brown, tea or even soy sauce urine, 6(27.3%) had acute renal injury (AKI), and 4(18.2%) had multiple organ dysfunction syndrome (MODS). After the treatment, 21 cases (95.5%) got better, and one case were discharged. All the patients with AKI were survived, three of them were treated by hemodialysis, and the other recovered gradually after massive fluid replacement.@*Conclusion@#Acute poisoning combined with RM is not uncommon in clinic. We should pay attention to examination of serum enzymes and other indicators, observe the clinical symptoms and make early diagnosis. The key to diagnosis and treatment is early fluid resuscitation, comprehensive treatment, blood purification and maintain the stability of water and electrolyte.

Comparison of different scoring systems in prognosis evaluation of acute poisoning / 中华劳动卫生职业病杂志

Objective@#To compare the predictive value of PSS, APACHEII, SAPSII and SOFA in the prognosis evaluation of acute poisoning.@*Methods@#Clinical data (including PSS score, APACHEII score, SAPSII score and SOFA score, within 24 hours after admission) of 231 acute poisoning patients admitted to the emergency intensive care unit EICU of our hospital from January 2015 to October 2016 was retrospectively analyzed. The patients were divided into the survival group and the dead group according to the 28-day clinical outcomes, comparing the differences of clinical data in each group. To analyze the correlation between PSS score, APACHEII score, SAPSII score and SOFA score in each group, comparing the value and the area under the ROC curve of four scoring systems and evaluate the predictive value of the four scoring systems.@*Results@#Comparing with the survival group and the dead group, PSS score, APACHEII score, SAPSII score and SOFA score were significantly different (P<0.01) . PSS score, APACHEII score, SAPSII score and SOFA score were significantly positive correlation (P<0.01) , the area under the ROC curve (AUC) of the four scoring systems were 0.833, 0.887, 0.843 and 0.843 respectively. The area under the ROC curve (AUC) of APACHEII score was higher than PSS score, SAPSII score and SOFA score, the difference was statistically significant (z=2.351, 2.317, 2.217; P=0.019, 0.021, 0.027) , there was no significant difference in the area (AUC) between the three scoring curves (P>0.05) . The cutoff value (cut-off) , sensitivity, specificity and accuracy rates of PSS score, APACHEII score, SAPSII score and SOFA score were (2.5, 93.1%, 50.9%, 61.5%) , (14.5, 82.8%, 75.7%, 77.48%) , (31.5, 77.6%, 76.90%, 77.08%) , (5.5, 77.60%, 74.60%, 75.35%) .@*Conclusion@#PSS score, APACHEII score, SAPSII score and SOFA score can evaluate the prognosis of patients with acute poisoning, but the APACHEII score is better than the other three scoring systems in evaluating the prognosis for its evaluation ability and accuracy rate.

The protective effect of Xuebijing on paraquat-induced HK-2 cells apoptosis and the underlying mechanisms / 中华劳动卫生职业病杂志

Objective@#To investigate the protective effect and mechanism of Xuebijing (XBJ) on paraquat (PQ) -induced apoptosis in Human kidney cell line-2 (HK-2) cells.@*Methods@#Routinely cultured HK-2 cells, (1) Cell growth inhibition experiment after PQ and XBJ intervention: PQ was divided into 0、200、400、800、1600 and 3200 μmol/L PQ groups, and the cell survival rate was detected after intervening 24、48 and 72 h. XBJ was divided into 0、5、10、20、40 mg/ml XBJ groups, and the cell survival rate was detected after intervening 24、48 and 72 h.To determine the rational drug concentration and the duration of action of XBJ and PQ. (2) PQ-induced HK-2 cell growth inhibition experiment antagonized by XBJ: The cells were divided into normal control group, PQ group (800 μmol/L) and PQ+XBJ group (The cells were pretreated with 5、10 and 20 mg/ml XBJ for 1 h, then cultured with PQ of 800 μmol/L) , After cultured 24 h、48 h and 72 h separately, the cell survival rate was detected. (3) HK-2 cells were divided into normal control group、PQ group (800 μmol/L PQ cultured for 24 h) 、PQ+XBJ group (pretreated with 10 mg/ml XBJ for 1 h, and then 800 μmol/L PQ cultured for 24 h) and XBJ group (10 mg/ml XBJ cultured 24 h). The apoptosis of cells was detected by flow cytometry. The protein expression of Bcl-2 and BAX in each group was detected by Western blotting. The expressions of caspase-3 and caspase-9 were detected by caspase-3 and caspase-9 activity kit active.@*Results@#(1) PQ could significantly reduced the survival rate of HK-2 cells and showed time and concentration dependence. The survival rate of HK-2 cells was about 55% after 800 μmol/L PQ contacted 24 h, XBJ under 20 mg/ml was no significant effect on the survival rate of HK-2 cells after cultured 72 h. (2) Compared with the PQ group, the survival rate of HK-2 cells of PQ+XBJ group was significantly increased (P<0.05). (3) Compared with the normal control group, the cell apoptosis rate of PQ group was significantly increased (P<0.05). Compared with the PQ group, the cell apoptosis rate of PQ+XBJ group was significantly decreased (P<0.05). (4) Compared with the normal control group, Bcl-2 protein expression in PQ group was significantly decreased and BAX protein expression in PQ group was significantly increased (P<0.05) ; compared with PQ group, Bcl-2 protein expression in PQ+XBJ group was significantly increased, BAX protein expression in PQ+XBJ group was significantly decreased (P<0.05). (5) Compared with the normal control group, the activities of caspase-3 and caspase-9 in PQ group were significantly increased (P<0.05). Compared with PQ group, the activities of caspase-3 and caspase-9 in PQ+XBJ group were decreased significantly (P<0.05) .@*Conclusion@#XBJ (10 mg/ml) has obvious protective effect on HK-2 cell injuried by PQ (800 μmol/L) , It can improve the survival rate of cells through reducing the apoptosis of HK-2 cells which induced by PQ.

Protective effects of histone deacetylase inhibitors trichostatin A on acute lung injury in septic mice / 中华急诊医学杂志

Objective To investigate the effects of histone deacetylase inhibitors trichostatin A (TSA) on acute lung injury in septic mice.Methods Septic mice model was induced by cecal ligation and puncture(CLP).Ninty male BALB/c mice of clean grade were randomly(random number) divided into six groups(n=15),namely sham operation group,CLP group,CLP+DMSO group,CLP+TSA 1 mg group,CLP+TSA 5 mg group,and CLP+TSA 10 mg group.TSA(1 mg/kg,5 mg/kg,10 mg/kg) was administrated 12 hours before operation by intraperitoneal injection.And mice in sham group were only treated with laparotomy without CLP,and 24 h later,all survived mice were sacrificed to obtain specimens.ELISA method was employed to detect the concentrations of TNF-α and IL-1β in BALF.The lung wet/dry ratio was calculated.Histopathology changes of lung tissues were observed under light microscope.Lung tissue cell apoptosis was detected by TUNEL method.Caspase-3,Caspase-9 and CytC were assayed by Western blotting.The survival rate of mice in each group was calculated by additional 120 mice.Data were analyzed by SPSS 23.0.Statistical analyses were performed using independent sample t-test to compare between two groups or one-way analysis of variance test to compare among muhiple groups.The survival rate of mice was analyzed by univariate analysis using log-rank test.Results The lung W/D(P=0.021),the concentrations of TNF-α(P=0.000 1)and IL-1β(P=0.000 6)in BALF,puhnonary pathological change(P=0.001 6),lung tissue cell apoptotic index(P=0.000 9),the levels of apoptosis proteins (P＜0.05) in CLP group were higher than those in sham group,while survival rate (P=0.000 1) in CLP groups was lower than that in sham group.Compared with DMSO,the TSA significantly reduced the lung W/D,the levels of TNF-α.IL-1β in BALF,pathologic changes of lung tissue,lung tissue cell apoptotic index and the levels of apoptosis proteins in septic mice(P＜0.05).The increase in survival rate (P=0.007 2) associated with TSA(10 mg/kg)administration.Conclusion TSA exerts protective effects through attenuating pro-inflammatory cytokines and lung tissue cell apoptosis in sepsis induced acute lung injury in mice.

The effects of miR-10a on the immune function of splenic CD4+CD25+ Treg cells in septic mice / 中华急诊医学杂志

Objective To investigate the role of miR-10a in CD4+CD25+Treg-mediated immunosuppression during sepsis and its potential role in immunotherapy for sepsis.Methods Sepsis mouse model was established by cecal ligation and puncture(CLP).Balb/c mice of clean grade were sacrificed 1,3,5,and 7 days after operation.Blood as well as spleen samples were harvested at given intervals.The splenic CD4+CD25+Treg cells and CD4+T cells were isolated by MACS microbeads.Cells were cultured,and phenotypes were analyzed by flow cytometry.The miR-10a expressed in Treg cells were detected by Real-time PCR.After administration of LV-mmu-miR-10a-5p-inhibition,the immunosuppressive function have been detected.Statistical analyses were performed using one-way analysis of variance (SPSS 19.0,Chicago,USA) test followed by Dunnett-t test to compare among three or more groups or by Student's t-test to compare between two groups.Results The percentages of splenic Tregs (CD4+CD25+/CD4+T) was (7.34±1.2)％ in normal group,and the increase in percentage of Tregs in spleen has been observed in septic mice (P＜0.05).The mean fluorescence intensity (MFI) of Foxp3+Treg was increased in septic mice compared with sham group (P＜0.05).The expression of miR-10a was significantly elevated on CLP 1-7 day (P＜0.05).After down-regulation of miR-10a in septic mice,the percentages of Tregs (CD4+CD25+/CD4+T) was significantly increased in septic mice (P＜0.05),the MFI of Foxp3+Treg was increased in septic mice compared with control group (P＜0.05).The CD4+T cell proliferative activity in CLP-induced mice was significantly suppressed on CLP 3 day compared with sham group (P＜0.05).After down-regulation of miR-10a in septic mice,the CD4+T cell proliferative activity was significantly suppressed compared with control group (P＜0.05).Conclusions Treg plays a critical role in immunosuppression in septic mice.Inhibition of miR-10a in vivo could enhence immunesuppression of CD4+CD25+Treg.Therefore miR-10a may participate in the regulation of CD4+CD25+Treg immunosuppression in sepsis and become the target for immunotherapy.

Mono-carbonyl analogues of curcumin prevents paraquat-induced apoptosis in HK-2 cell line by inhibiting oxidative damage and inflammation / 中华劳动卫生职业病杂志

Objective@#To investigate the effects of mono-carbonyl analogues of curcumin (L6H21) on paraquat (PQ) -induced injury in HK-2 cell line and explore its underlying mechanisms.@*Methods@#Cultured HK-2 cells were challenged by PQ with or without L6H21 treatment. Cell viability and apoptosis were determined by CCK-8 assay and flow cytometry, respectively. Gene expressions and protein levels of apoptotic and inflammatory factors were assessed by RT-PCR, ELISA, and western blot. Intracellular ROS production was detected by DCFH-DA staining. Superoxide dismutase (SOD) and malondialdehyde (MDA) were examined by chemical colorimetry.@*Results@#1) PQ challenge significantly inhibited HK-2 cells proliferation, which was prevented by L6H21 administration. PQ dramatically induced HK-2 apoptosis evidenced by increasing expressions of caspase-9, caspase-3 and Bax, while decreasing Bcl-2 level. However, PQ induced these apoptotic effects in HK-2 cells were reversed by L6H21. Similarly, PQ exposure obviously enhanced activity of NF-κB and levels of cytokines (TNF-α、IL-6) in HK-2 cells, which was inhibited by L6H21. Furthermore, administration of L6H21 inhibited PQ induced ROS and MDA production, and promoted SOD level in HK-2 cells.@*Conclusion@#L6H21 administration inhibits PQ-induced apoptosis in HK-2 cells possibly by reducing inflammation and oxidative damage.

Analyses on relevant factors of the prognosis of elderly patients with acute poisoning / 中华劳动卫生职业病杂志

Objective@#To explore the risk factors influencing the prognosis of elderly patients with acute poisoning.@*Methods@#We retrospected 177 elderly patients with Acute Poisoning who were treated in the emergency department of the first affiliated hospital of wenzhou medical university from July 2009 to May 2015. According to the outcome of patients, we distributed the patients to death group (31 cases) and survival group (146 cases) , compared the clinic data and using multivariate analysis with Logistic regression to prognosis factors.@*Results@#There were 177 cases in total, with 146 survivors (82.5%) and 31 deaths (17.5%) . In which 102 cases (57.6%) had chronic underlying diseases. There were 28 cases of pesticide poisoning in the death group, and the fatality rate of pesticide poisoning was 23.5%. The mortality rate was 12.8% in the 60-69 years-old group (11/86) , 20% (13/65) in the 70-79 years-old group, 26.9% (7/26) in the 80-89 years-old group. The most common reason of poisoning was intentional ingestion, with 100 cases (56.5%) . The tract of the poisoning was mainly in digestive system, including 148 cases (83.6%) . The PSS score and APACHE-II score were 2.97±0.18 and 19.8±2.8 in the death group, 2.27±0.81 and 12.8±5.3 in the survival group. Compared with the survival group, poison (pesticides or non) 、poisoning route、cause of poisoning、PSS score、APACHEⅡ score have significant difference in death group (P<0.05) . Poison (pesticides or non) 、PSS score、APACHEⅡ, were the independent risk factors of poor prognosis.@*Conclusion@#Most of the elderly patients with acute poisoning have one or more chronic underlying diseases, the digestive tract ingestion and pesticide poisoning are more common. The fatality rate of the old patients is significantly higher than that of non elderly poisoning. Type of toxications, PSS score and APACHE-II score are the prognostic factors in elderly patients.

Value of base excess in predicting the prognosis of patients with paraquat poisoning / 中华劳动卫生职业病杂志

Objective@#To explore prognostic factors of acute paraquat poisoning (APP) , analyze the correlation between base excess (BE) and plasma concentration of paraquat (C-PQ) and discuss BE level in evalua-tion of prognosis of acute paraquat poisoning patients.@*Methods@#We retrospectively selected 84 APP patients who admitted to Emergency Intensive Care Unit (EICU) of our hospital from 2009.9.1 to 2015.8.31.Clinical data from 84 APP patients were analyzed. BE、C-PQ、time of gastric lavagesince ingestion、time of hemoperfusion since ingestion、severity index of paraquat poisoning (SIPP) 、white blood cell (WBC) 、percentage of neutrophils (N%) 、hemoglobin (HB) 、creatinine (Cr) 、alanine aminotransferase (ALT) 、aspartate aminotransferase (AST) 、partial pressure of oxygen (PaO2) 、partial pressure of carbon dioxide (PaCO2) and other laboratory parameters were measured. A total of 41 patients in non-survivors died during the 30 days after admission and 43 patients in survivors survived during the 30 days. The factors of prognosis in paraquat poisoining and the role of BE in evalu-ating prognosis was analyzed, as well as the correlation between BE and C-PQ.@*Results@#1.Logistic regression analyses showed BE、C-PQ、ALT、AST、Cr was of prognostic significance[odds ratio (OR) of BE: 0.511, 95%CI 0.267, 0.978; C-PQ:-=0.999, 95%CI 0.999, 1.000; both P<0.05] ; 2.The area under the receiver operating characteristic curve (ROC curve) of BE、C-PQ and prognosis were 0.775、0.927 respectively, BE≤-1.7 mmol/L was the best cut-off value, the sensitivity、specificity for predicting were 82.9%、62.8%, the evaluation value was lower to C-PQ>3 273.935 ng/ml (AUC 0.927, 78.0%、95.3%) ; 3.BE negative correlated with C-PQ[-1.100 (-4.100, -0.200) , -5.900(-8.650, -2.500) , both P<0.05]. (r=-0.4, P<0.01).@*Conclusion@#These results suggest that BE may be useful for the prediction of prognosis in PQ poisoning and BE negative correlated with C-PQ.

The trend of changes in expression of Klotho and autophagy in sepsis-induced acute kidney injury mice model / 中华急诊医学杂志

Objective To investigate the relationship between Klotho and autophagy in sepsisinduced acute kidney injury mice model.Methods The male healthy Balb/c mice were used to establish the model of sepsis-induced acute kidney injury by using cecal ligation and puncture (CLP).Mice were sacrificed at 3 h,6 h,12 h,1 d,2 d,3 d,and 5 d after CLP (n =12 for each interval) and on 1 d 6 mice in sham group as well as 6 mice in normal group were sacrificed at the same time.Scr and BUN in the blood serum were detected.The HE and PAS staining were employed for observation on the histopathological changes in kidney tissues under light microscope.The autophagosomes were observed under transmission electron microscope (TEM).The renal protein of Klotho,LC3 and P62 were detected by using Western blot and Immunohistochemistry.Statistical analyses were performed using Student's t-test by SPSS 23.0.software.Results Scr and BUN increased significantly after CLP,especially on 1 d,respectively (165.64 ± 20.56) μmol/L and (45.51 ± 4.05) mmol/L.HE and PAS staining showed renal tissue was damaged obviously 1 d after CLP,as indicated by desquamation of the brush border of proximal tubular epithelial cells,appearance of bare basement membrane,and interstitial inflammatory cell infiltration.Under TEM,autophagosomes and phagocytosis were observed.Compared with sham group,the expression of Klotho protein decreased gradually from 3 h to 1 d and dropped to the trough at 1 d (t =51.851,P ＜0.01),then resumed gradually from 2 d to 5 d.On the contrary,the activation of autophagy increased as indicated by the expression of LC3-Ⅱ/L3-Ⅰ and p62.Autophagy was induced gradually from 3 h to 1 d and reached peak at 1 d,then declined gradually from 2 d to 5 d (P ＜ 0.01).The protein of Klotho and LC3-Ⅱ mainly distributed in renal tubular cytoplasm,and Klotho was reduced significantly (t =-8.371,P ＜ 0.01) and LC3-Ⅱ appeared in high density remarkably (t =4.995,P =0.001) on 1 d after CLP.Conclusions Klotho protein reduction and autophagy protein increase were observed in sepsis-induced acute kidney injury,and the expressions of Klotho and autophagy acted out in certain extent of time dependence.

The protective effect of bone marrow mesenchymal stem cells carrying antioxidant gene superoxide dismutase on paraquat lung injury in mice / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the possible mechanism and protective effect of BMSCs (bone mesenchymal stem cells) carrying superoxide dismutase (SOD) gene on mice with paraquat-induced acute lung injury.</p><p><b>METHODS</b>To establish the cell line of BMSCs bringing SOD gene, lentiviral vector bringing SOD gene was built and co-cultured with BMSCs. A total of 100 BALB/c mice were randomly divided into five groups, namely Control group, poisoning group (PQ group) , BMSCs therapy group (BMSC group) , BMSCs-Cherry therapy group (BMSC-Cherry group) , BMSCs-SOD therapy group (BMSC-SOD group) . PQ poisoning model was produced by stomach lavaged once with 1 ml of 25 mg/kg PQ solution, and the equal volume of normal saline (NS) was given to Control group mice instead of PQ. The corresponding BMSCs therapy cell lines were delivered to mice through the tail vein of mice 4h after PQ treatment.Five mice of each group were sacrificed 3 d, 7 d, 14 d and 21 days after corresponding BMSCs therapy cell lines administration, and lung tissues of mice were taken to make sections for histological analysis. The serum levels of glutathione (GSH) , malondialdehyde (MDA) , SOD, and the levels of transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in lung tissue were determined. The level of SOD was assayed by Westen-blot.</p><p><b>RESULTS</b>Compared with Control group, the early (3 days) levels of SOD protein in lung tissue of PQ group obviously decreased, and the late (21 days) levels of SOD obviously increased, while in therapy groups, that was higher than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Compared with Control group, the levels of plasma GSH and SOD of PQ group and each therapy group wae significantly lower than those in Control group, while in therapy groups, those were higher than those of PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) .Compared with Control group, the level of plasma MDA, TNF-α and TGF-β in PQ group and therapy groups were significantly higher, while in therapy groups, that was lower than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Lung biopsy showed that, the degree of lung tissue damage in each therapy group obviously reduced.</p><p><b>CONCLUSION</b>SOD is the key factor of the removal of reactive oxygen species (ROS) in cells, that can obviously inhibit the oxidative stress damage and the apoptosis induced by PQ, thus significantly increasing alveolar epithelial cell ability to fight outside harmful environment.</p>

The effect of resveratrol on paraquat-induced acute lung injury in mice and its mechanism / 中华危重病急救医学

Objective To investigate the effect of resveratrol (Res) on paraquat (PQ)-induced acute lung injury (ALI) and mortality in mice and the mechanism of nuclear factor-κB (NF-κB) inflammatory pathway. Methods Sixty-eight healthy male ICR mice with grade SPF were enrolled, among them 20 mice were used for mortality observation (n = 10), and other 48 were used for determination of related parameters (n = 6). The mice were randomly divided into four group s: normal saline (NS) control group, Res control group, PQ group and PQ + Res group. The mice in the latter two groups were subdivided into 6, 24, 72 hours subgroups. The PQ poisoning model of mice was reproduced by one injection of 30 mg/kg PQ intraperitoneally. The mice in PQ + Res group were given 60 mg/kg Res intraperitoneally on the contralateral side after PQ injection. The mice were sacrificed at 6, 24, 72 hours after PQ poisoning, and lung tissue was harvested. The serum levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6 and IL-1β) were determined by enzyme linked immunosorbent assay (ELISA). The pathological changes in lung tissue were observed with electron microscopy. Apoptosis cells in the lung were identified by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) for the estimation of apoptosis rate. The protein expression of NF-κB p65 was determined by Western Blot. Results Compared with PQ group, the death number of mice at 48, 72, 96 hours in PQ + Res group was slightly decreased (0 vs. 2, 2 vs. 5, 4 vs. 6) but without statistically significant difference (all P > 0.05). Under electron microscope, the lung injury in PQ group was severer than that in NS control group, and Res was found to be able to alleviate the lung injury. Compared with NS control group [(2.45±0.61)%], the apoptosis rate at 6 hours in PQ group was significantly increased [(8.42±1.48)%], and peaked at 72 hours [(21.23±3.47)%]. Res could decrease the apoptosis rate after PQ poisoning [6 hours: (5.56±1.31)% vs. (8.42±1.48)%, 24 hours: (11.14±2.07)% vs. (16.88±2.96)%, 72 hours: (13.28±2.32)% vs. (21.23±3.47)%, all P < 0.05]. The serum levels of TNF-α, IL-6, and IL-1β, and NF-κB p65 in lung tissue were all markedly increased after PQ poisoning, and they were significantly decreased after Res intervention as compared with those of PQ group [TNF-α (ng/L): 2.62±0.29 vs. 4.06±0.74 at 6 hours, 3.98±0.41 vs. 6.79±0.80 at 24 hours, 5.06±0.75 vs. 11.00±0.75 at 72 hours; IL-6 (ng/L): 14.19±1.54 vs. 16.55±1.24 at 6 hours, 13.21±1.37 vs. 19.73±0.85 at 24 hours, 13.72±0.56 vs. 22.45±0.72 at 72 hours; IL-1β (ng/L): 8.54±1.64 vs. 12.59±0.66 at 6 hours, 10.15±0.29 vs. 16.24±1.03 at 24 hours, 16.14±0.70 vs. 19.55±0.56 at 72 hours; 6-hour NF-κB p65: (1.34±0.07) folds vs. (1.86±0.11) folds when the expression in NS control group was represented as 1, all P < 0.05]. Conclusions Res cannot lower the mortality in mice with PQ poisoning, but it seems to be able to attenuate PQ-induced ALI and cell apoptosis. The mechanism responsible for the latter maybe the inhibitive effect of Res on NF-κB p65 translocation and cytokines production.