Objective To clinically validate the precision of diagnostic Sepsis-3 criteria, and to guide and generalize its clinical application.Methods A multicenter retrospective observational study was conducted. The patients admitted to intensive care unit (ICU) of 6 tertiary hospitals in Zhejiang Province from January to June 2015 were enrolled, and the patients satisfying the diagnostic criteria of Sepsis-2 and Sepsis-3 were screened. Population characteristics between the patients satisfying two editions were compared, and the diagnosis accuracy rate in different degree hospitals were investigated. According to the doctor's diagnosis, the patients who met the criteria of Sepsis-2 were divided into diagnosis group and non-diagnosis group, and the factors influencing the diagnosis of sepsis were analyzed by logistic regression. The patients meeting Sepsis-2 but no meeting Sepsis-3 were served as exclusion group, and those meeting Sepsis-2 and Sepsis-3 were served as enroll group, and the characteristics of patients between the two groups were compared. Receiver operating characteristic (ROC) curve was plotted to evaluate the predictive value of systemic inflammatory response syndrome (SIRS) score, sepsis-related quick sequential organ failure assessment (qSOFA) and sequential organ failure assessment (SOFA) on death, and whether the consistency of qSOFA and SOFA would affect the sensitivity of definition. The patients meeting Sepsis-2 were divided into non-survived group and survived group, and the factors associated with death were analyzed by logistic regression.Results Finally, 1423 patients were enrolled, 3 patients with age < 18 years and 19 patients with missing data were excluded. There were 363 patients and 329 patients met Sepsis-2 and Sepsis-3, respectively. No significant differences were found in populationcharacteristics between the groups of Sepsis-2 and Sepsis-3 (allP > 0.05) except for acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score [19.10 (8.00) vs. 20.28 (8.00),P < 0.05]. It was shown on the clinical data analysis of the hospitals that the patients meeting Sepsis-2 and Sepsis-3 in hospital 3 had the highest 28-day mortality (60.4% and 60.0%) with the lowest rate of diagnosis (0). While in the hospital 1, the patients had the lowest 28-day mortality (22.9% and 27.2%), and the rate of diagnosis was 19.5%. Interestingly, the patients in hospital 4 had the highest diagnosis rate of sepsis (44.8%), but 28-day mortality was both 58.6%. It was shown by logistic regression analysis that the patients with old age [odds ratio (OR) = 0.970,P = 0.021], high blood lactate (OR= 0.443,P = 0.004), high blood pressure (OR = 0.957,P = 0.009) and low oxygenation index (OR = 1.004,P = 0.024) were easy to neglect diagnosis. Compared with Sepsis-3 exclude group, the patients in Sepsis-3 enroll group were older [years: 68.52 (26.00) vs. 53.75 (18.00),P < 0.01] with higher APACHEⅡ score [20.38 (8.00) vs. 7.72 (6.00),P < 0.01], higher blood lactate [mmol/L: 3.45 (3.00) vs. 1.95 (1.20), P > 0.05], longer length of ICU stay [days: 22.42 (22.00) vs. 15.13 (16.00),P < 0.01], and higher 28-day mortality [45.29% (149/329) vs. 14.71% (5/34),P < 0.01], indicating that the diagnostic efficiency of Sepsis-2 was low, the diagnostic specificity of Sepsis-3 was high, and the prognosis of Sepsis-3 patients was worse. It was shown by ROC curve analysis that the prognostic value of SIRS, qSOFA and SOFA to mortality was gradually increased [area under ROC curve (AUC) was 0.567, 0.597, 0.683, respectively], but the prognostic value were all low. Comparing patients meeting qSOFA and (or) SOFA in Sepsis-2, significant differences were found in APACHE Ⅱ score [17.55 (7.00) vs. 23.24 (8.00)] and 28-day mortality [38.75% (31/80) vs. 58.59% (75/128), bothP < 0.01]. The patients who just met the qSOFA or SOFA, their 28-day mortality was up to 38.75%, suggesting that qSOFA should not be ignored. Compared with survived group, the patients in survived group were older with higher APACHE Ⅱ score and shorter length of ICU stay (allP < 0.05). It was shown by logistic regression analysis that APACHE Ⅱ score (OR = 1.199,P = 0.000) and length of ICU stay (OR = 0.949,P = 0.000) were related with death.Conclusion Patients satisfied Sepsis-3 were easier to develop more organ failure, Sepsis-3 and higher death prediction than Sepsis-2 and higher diagnosis specificity, but data shows that there is extra room for improvement.

Objective To explore effect of immune function of children with EB virus infectious mononu-cleosis (IM). Methods From Sep. 2011 to Jun. 2013, a total of 62 cases of children with IM were analyzed retrospectively. The proportion of T-lymphocyte subsets ,B-lymphocytesand and NK cells in EBV-DNA group with positive and negative were compared. Results The numbers of IM EBV-DNA-negative cases were 18 (29.03%), the numbers of IM EBV-DNA-positive cases were 44(70.97%), the averages of EBV-DNA in positive group is 6.1 × 103/mL. CD3+,CD8+cell ratios in IM children are significantly higher than control group (P<0.01), CD4+, CD19+cell ratios are significantly lower than control group (P<0.01), NK cells are not significantly different (P>0.05). In the IM children, CD4+, CD19+cells in DNA-positive group are significantly less than DNA-negative group (P<0.01), CD8+cells are higher than DNA-negative group (P<0.05), NK cells are not significantly different (P>0.05). Conclusions Positive blood EBV-DNA is closely related to changes in T-lymphocyte and B-lymphocyte, but isn′t obviously related in NK cells. So effect of EB virus in IM child have certain clinical values to judge the severity and prognosis.

3T3-L1 adipocytes were cultured and differentiated into mature adipocytes in vitro. The adipocytes were intervened by ACAT inhibitor( 2 μg/ml) and ox-LDL with various concentrations (0,25,50,75,and 100 μg/ml)for 48 h,ACAT inhibitor( 2 μg/ml) and ox-LDL( 50 μg/ml) at the 0,6,18,36,and 48 h,or ACAT inhibitor( 2 μg/ml),ox-LDL( 50 μg/ml),and TUDCA with various concentrations(0,100,200,and 400 μ mol/L)for 48 h,respectively.The levels of visfatin in supernatant were examined by ELISA and the expressions of protein GRP78 and CHOP in adipocytes were detected by Western blot.After the adipocytes were treated with ACAT inhibitor and ox-LDL at different concentrations for 48 h,the cholesterol concentration and the expressions of GRP78 and CHOP protein in adipocytes and the visfatin levels in the supernatant fluid were increased with the increase of the ox-LDL concentration.The differences had statistical significance in the experimental groups compared with blank control group( all P＜0.05 ).After the intervention with ACAT inhibitor and ox-LDL for different durations,the expressions of GRP78 and CHOP protein in adipocytes and the visfatin levels in the supernatant fluid were up-regulated in a timedependent manner.The differences between experimental groups and blank control had statistical significance( all P＜0.05 ).After the intervention with ACAT inhibitor,ox-LDL,and different concentrations of TUDCA for 48 hours,the expressions of GRP78 and CHOP protein in adipocytes and visfatin levels in the supernatant fluid were down-regulated in a dose-dependent manner and as compared with blank control group the difference were statistically significant( all P＜ 0.05 ).The increase of cholesterol load in adipocyte may promote the visfatin secretion,denoting that the mechanism might be due to the enhancement of endoplasmic reticulum stress in aidpocytes.

Objective To investigate the regulation on extracellular signal regulated kinase (ERK )1/2 and p38 signaling pathways and immune expression by moist exposed burn therapy/moist exposed burn ointment(MEBT/MEBO)in diabetic foot and their relationship ,to explore the repair mechanism of MEBT/MEBO on diabetic foot ulcers .Methods Totally 40 diabetic foot pa‐tients were treated by MEBT/MEBO ,to take wound tissue before and after treatment and detect the expression of ERK 1/2 ,p38 , MAPKK6 ,c‐myc ,Akt ,ATF2 ,IgA ,IgM ,IgG ,C3c and C4c by immunohistochemistry ,to investigate their relationship .Results Af‐ter treatment with MEBT/MEBO ,the area of foot wounds in 39 patients was reduced in different degree .Only one patient had no obvious change .14 patients(35 .00% ) were markedly effective ,25 patients(62 .50% ) and 1 patient(2 .50% ) were ineffective .Before andaftertreatment,allpositiveexpression,positiveimmunoreactivity(anyindex)andpositiveexpression(specificindex)ofsignal pathway molecules were statistically significant (P<0 .05) .While the positive rate of molecular expression in wound pathway in‐creased ,the positive expression rate of immune factor increased .Before treatment ,a small amount of immune factors were found in the wound tissue .After treatment ,the immune factors IgA ,IgM ,IgG ,C3c ,C4c were distributed widely and diffusely .Before treat‐ment ,the wound tissue showed a very small number of signal molecules .After treatment ,the signal pathway molecules MEBT/MEBO and p38 ,MAPKK6 ,c‐myc ,Akt ,ATF2 showed broad and diffuse distribution .Conclusion MEBT/MEBO may promote the expression of ERK1/2 and p38 signaling molecules and immune in diabetic foot ,p38 and ERK1/2 signaling pathway may promote the healing of diabetic foot wound by increasing the expression of immune .

Objective To investigate the effect of lidocaine on Staphylococcus aureus exotoxin-stimulated peripheral blood mononuclear cells (PBMCs) from patients with atopic dermatitis (AD).Methods Peripheral blood samples were collected from 6 patients with AD,and PBMCs were isolated by a routine method.Then,the PBMCs were stimulated by the Staphylococcus aureus exotoxin toxic shock syndrome toxin-1 (TSST-1) in the absence or presence of lidocaine at varying concentrations.The 3H-TdR incorporation method was performed to detect the proliferation of monocytes,and enzyme-linked immunosorbent assay (ELISA) to quantify the levels of T helper type 1 (Th1) and Th2 cytokines released by PBMCs.Human HaCaT keratinocytes were co-cultured with lidocaine-and TSST-1-stimulated PBMCs from patients with AD for 72 hours,then,Western blot was conducted to examine the expression of filaggrin protein in HaCaT cells.Results TSST-1 (100 μg/L) significantly enhanced the proliferation of PBMCs from patients with AD (stimulation index =75 ± 2.12,P ＜ 0.05),as well as the release of tumor necrosis factor-α (TNF-α),interferon (IFN)-γ,interleukin (IL)-2,IL-12,IL-4,IL-5 and IL-13 by the PBMCs (all P ＜ 0.05).Compared with the blank control group,100 μmol/L lidocaine significantly inhibited the TSST-1-stimulated proliferation of PBMCs from patients with AD (stimulation index =58 ± 3.14,P＜ 0.05),as well as the release of IL-4,IL-5,IL-13,TNF-α and IFN-γ by the stimulated PBMCs (all P ＜ 0.05).Western blot showed that 100 μmol/L lidocaine significantly blocked the down-regulation of filaggrin expression in HaCaT cells (P ＜ 0.01).Conclusion Lidocaine has a significant inhibitory effect on the activation of TSST-1-stimulated PBMCs from patients with AD.

Chronic lung diseases, including bronchial asthma, chronic obstructive pulmonary disease (COPD), chronic bronchitis, allergic rhinitis and repeated respiratory tract infection (RRTL) in infants, exacerbate frequently in winter because of respiratory viral infections and low temperature. Summer acupoint application therapy (SAAT) is thought to be effective in reducing exacerbation frequency of chronic lung diseases in winter. It is a kind of therapy using a herbal mixture for external application on special acupoints during summer. The herbal mixture basically contains Semen Sinapis Albae, Herba Asari, Radix Euphorbiae Kansui and Rhizoma Corydalis. The acupoints include Feishu (BL13), Dazhui (GV14) and Danzhong (CV17). Through a large-scale multicenter trial based on three years of clinical observation, and retrospective and prospective analyses, this study aims to explore the efficacy of SAAT.

Objective:To investigate the effects of Xiaoyu-Jiangzhi capsules on blood lipid, carotid artery atherosclerosis (CAA) and plaque in apolipoprotein E knockout (ApoE -/-) mice. Methods:The ApoE -/- mice were fed with high-fat food to establish carotid atherosclerosis model. The ApoE -/- mice were randomly by weight divided into model group, Atorvastatin group, low- and high-dose Xiaoyu-Jiangzhi capsules group. The C57BL/6cnc mice were used as control group and fed with normal diet. The Atorvastatin group was given atorvastatin suspension 1.3 mg/kg, low and high dose groups were given Xiaoyu-Jiangzhi capsule suspension 325 and 975 mg/kg, and the control group and model group were given equal volume of distilled water. The mice were gavaged with 0.1 ml/10 g body weight, once a day, and the weight of mice was recorded weekly. After 12 weeks of continuous intragastric administration, the blood lipid and liver /body weight index of the mice were measured. Carotid arteries were sliced to conduct oil red O staining and VG staining for the pathological analysis. Results:After 12 weeks of drug administration, the weight of mice in the high-dose group was significantly lower than the model group. The level of TC (25.92 ± 4.21 mmol/L vs. 30.39 ± 4.67 mmol/L) and LDL-C (7.97 ± 2.14 mmol/L vs. 10.26 ± 1.97 mmol/L) in the high-dose group significantly decreased ( P<0.05), the level of HDL-C in the low and high-dose group significantly increased ( P<0.05). The pathological results showed that after 12 weeks of administration, the carotid artery lipid deposition blockage rate in the Atorvastatin group and the high dose group were significantly smaller than the model group( P<0.05), and no vascular plaque has been formed. Conclusion:The Xiaoyu-Jiangzhi capsules could reduce LDL-C, increase HDL-C levels, reduce the constriction of arterial stenosis and slow down the formation process of carotid plaque.

OBJECTIVETo investigate the expression of full-length spleen tyrosine kinase [SYK (L)] mRNA and protein in human oral squamous cell carcinoma (OSCC) as well as its possible effects on the invasion and metastasis of OSCC.

METHODSThe expression of SYK (L) was detected in 27 cases of OSCC tissues and its matched adjacent non-cancerous tissues by real-time quantitative polymerase chain reaction (RT-qPCR), Western blot, and immunohistochemistry. Fourteen cases of normal oral gingival tissues were also analyzed as a normal control.

RESULTSReduced mRNA and protein expression of SYK (L) in OSCC tissues was observed compared with that in normal oral gingival tissues (P<0.01) and adjacent non-cancerous tissues (P<0.05). SYK(L) expression was significantly associated with lymph-node metastasis (P<0.05).

CONCLUSIONSYK(L) is a candidate tumor suppressor for OSCC tissues, and has an inhibitive effect on the initiation, proliferation, and lymph-node metastasis of human OSCC.

Blotting, Western ; Carcinoma, Squamous Cell ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Neoplasms ; metabolism ; RNA, Messenger ; Syk Kinase ; metabolism

Objective To validate of the Chinese system for cardiac operative risk evaluation (SinoSCORE) in Cantonese surgery patients. Methods Data from Guangdong Cardiovascular Institute in the period January 2004 through December 2008 were analyzed on 2462 Cantonese heart surgery patients. First, compared risk factors of this series and database of SinoSCORE, and then calculated the additive score of each patients and evaluate the discrimination and calibration of sinoSCORE in Cantonese patients. Results There were some differences between the risk factors of patients from two groups. The gender,smoking, diabetes, hyperlipemia, hypertension, chronic pulmonary diseases, stroke, cardiovascular surgery history, left main disease, atrial fibrillation/atrial flutter, pulmonary arterial hypertension, concomitant coronary surgery and concomitant valve surgery in Cantonese patients were different between two groups. However, The SinoSCORE was able to predict the in-hospital mortality of senior patients with good discrimination ( Hosmer-Lemeshow test, P = 0. 34 ) and calibration ( the area under the receiver operating characteristic curve, 0.84, P ＜ 0.01 ). Conclusion SinoSCORE was able to predict the in-hospital mortality of Cantonese heart surgery patients.

OBJECTIVETo investigate the role of histone acetylation in regulating influenza virus replicative intermediate double-stranded RNA (dsRNA)-induced interleukin-6 (IL-6) expression in A549 cells.

METHODSA549 cells were treated with influenza virus replicative intermediate dsRNA, histone deacetylase (HDAC) inhibitor trichostatin A (TSA), or HADC small interfering RNA (siRNA). The changes in the cellular IL-6 promoter activities were detected by dual-luciferase assay, and IL-6 mRNA and protein expressions in the cells were determined using real-time RT-PCR and ELISA, respectively.

RESULTSInfluenza virus replicative intermediate dsRNA obviously up-regulated IL-6 expression in the cells. HDAC inhibitor TSA significantly enhanced the activity of IL-6 promoter and increased IL-6 mRNA expression in A549 cells, and HDAC3 may play an important role in this process. HDAC inhibitor TSA and DNMT inhibitor DAC showed no synergic effect in regulating IL-6 expression.

CONCLUSIONSInfluenza virus replicative intermediate dsRNA-induced IL-6 expression in A549 cells is regulated by histone acetylation.

Acetylation ; Cell Line, Tumor ; Gene Expression Regulation ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Humans ; Interleukin-6 ; metabolism ; Orthomyxoviridae ; genetics ; metabolism ; Promoter Regions, Genetic ; RNA, Double-Stranded ; RNA, Messenger ; genetics ; RNA, Viral