Objective To observe the effect of severe heatstroke on intestinal mucosal barrier function,and explore its correlation with systemic inflammatory reaction.Methods The SPF male BALB/c mice were randomly divided into normal control group,40 ℃ heat stress group and 42 ℃ heat stress group,with 6 mice in each group.The mice in normal control group were observed at normal temperature with (25.0 ± 0.5)℃,and the mice in heat stress groups were challenged with a temperature of (35.5 ± 0.5) ℃ and a humidity of (60 ± 5)％ until body temperature increase up to 40 ℃ or 42 ℃ followed by recovering the surroundings temperature to normal temperature for 12 hours.The blood of medial angle of eye of mice in each group was collected for determination of plasma lipopolysaccharide (LPS),tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) levels,and diamine oxidase (DAO) activity with enzyme linked immunosorbent assay (ELISA).The level of D-lactic acid was determined with ultraviolet spectrophotometer.Then the mice in each group were sacrificed,and mesenteric lymph node (MLN),liver,spleen,lung,kidney tissues,and the blood from portal vein and caval vein were collected for colony count to observe the intestinal bacterial translocation.The ileum tissue was collected for observation of changes in histomorphology and ultrastructure of small intestine mucous membrane with microscope.Pearson linear regression analysis was used to explore the correlation between intestinal mucosal barrier dysfunction and systemic inflammatory response.Results Compared with normal control group,plasma LPS,inflammatory parameters such as TNF-α and IL-6,and gut barrier function parameters such as DAO and D-lactic acid levels as well as the rate of bacterial translocation after heat stress were significantly increased,and the differences were more obvious in 42 ℃ heat stress group [LPS (EU/L):740±50 vs.340±40,TNF-α (ng/L):148.06±36.61 vs.12.89 ± 1.67,IL-6 (ng/L):110.91 ± 9.97 vs.18.02 ± 2.20,DAO (U/L):1 760 ± 400 vs.670± 50,D-lactic acid (mg/L):9.60 ± 1.48 vs.5.08 ± 0.28,rate of bacterial translocation:78.6％ (33/42) vs.9.5％ (4/42),all P ＜ 0.01].It was shown by Pearson linear regression analysis that plasma LPS,TNF-α,IL-6 were positively correlated with DAO activity (r values were 0.834,0.808,0.836,respectively) and D-lactic acid (r values were 0.811,0.811,0.800,respectively) in 42 ℃ heat stress group (all P =0.000).It was showed by microscope that the changes in histomorphology and ultrastructure changes in intestinal mucosa were found after heat stress,and was obvious in 42 ℃ heat stress group as following:villus atrophy and falling off,infiltration of neutrophils and lymphocytes,the microvillus on the surface of mucosa cells were short and small,arranged in disorder,the tight junction between epithelial cells became widen,the mitochondrion and endoplasmic reticulum swelled obviously.Conclusion During the early stage of severe heatstroke,the damage of intestinal mucosal was obvious,and it has close correlation with systemic inflammatory response.

Objective: To preliminarily investigate the relationship between the baseline level of serum soluble ST2 (sST2) and 30-day MACE occurrence rate in patients with ST-elevation myocardial infarction (STEMI).
Methods: A total of 121 patients with confirmed diagnosis of STEMI in our hospital from 2015-05-01 to 2015-07-30 were consecutively enrolled. According to baseline sST2 level, the patients were divided into 2 groups:Low sST2 group, the patients with sST2≤56.68 ng/ml, n=61 and High sST2 group, the patients with sST2>56.68 ng/ml, n=60. Clinical condition and 30-day MACE (defined as death and new onset of congestive heart failure) occurrence rate were compared between 2 groups.
Results: ① The systolic blood pressure (SBP), Killip class≥II grade, blood levels of cTNI, NT-proBNP, hs-CRP and LVEF were different between 2 groups, all P<0.05. ② Baseline sST2 level was positively related to heart rate (r=0.271, P=0.003), Hs-CRP (r=0.359, P=0.000), cTNI (r=0.324, P=0.000) and NT-proBNP (r=0.425, P=0.000);negatively related to SBP (r=-0.226, P=0.013) and LVEF (r=-0.406, P=0.000).③30-day MACE occurrence rate was different between 2 groups (8.2%vs 30%, P=0.002). ④ Multivariate Cox regression analysis presented that sST2>56.68 ng/ml was the risk factor for 30-day MACE occurrence (HR=1.152, 95%CI 1.078-1.231, P=0.000).
Conclusion: Increased baseline level of sST2 implied the higher incidence of death and new onset of congestive heart failure in STEMI patients.

Objective To observe the oxidative stress, integrity of lysosome and apoptosis of intestinal epithelial cells 6 (IEC-6) after heat stress, and explore the pathogenesis of intestinal damage caused by heat stress.Methods In the heat stress groups,the cells were incubated at 43℃ for 1 hour, then, further incubated at 37℃ and 5% CO2 for 0, 1, 3, 6 and 12 hours respectively; in the medicine intervention group, the cells were pretreated with the medicine 1h before heat stress; while in control group, the cells were incubated at 37℃ and 5% CO2. The amount of reactive oxygen species (ROS) was assayed with 2', 7'-dichlorofluorescin diacetate (DCFH-DA) and dihydroethidium (DHE) staining. The stability of lysosome membrane was checked by AO staining. Apoptosis was analyzed by flow cytometry using annexinⅤ-FITC/PI staining, CCK-8 assay was used to assess cellular viability.Results Compared with control group, cell viability decreased and apoptosis increased at 1 h after heat stress, which was the most obvious at 12h after rewarming (P<0.05). While ROS and pale cells increased immediately after heat stress and the increase become the most obvious (P<0.05). The cell viability in E-64 pretreatment group was significantly improved such as apoptosis reduction, compared with heat stress group (P<0.05).Conclusion Heat stress could induce robust increase of ROS, which mediates lysosome damage and results in cell apoptosis, thus suggesting that ROS-lysosome pathway may play an important role in intestinal injury in heat stress.

Objective To explore the protective effect of propofol on endothelial cells during heat stress and its protective effect to mitochondra. Methods Heat stress model of human umbilical vein endothelial cell was established when cells were incubated at 43℃ for 2h, then further incubted at 37℃, 5%CO2 for 6h. The experimental group was subdivided into six groups, including 37℃ group, 37℃ plus intralipid group (negative control group), 37℃ plus propofol group, 43℃ plus propofol group, 43℃ plus intralipid group, H2O2 plus propofol group (positive control group); Pretreated with 50μmol/L propofol, 0.2ml intralipid or 25μmol/L H2O2 before heat stress at 43℃, while the cells in the control group were incubated at 37℃. Cell viability was tested by CCK-8. ROS, mitochondrial membrane potential and the changes in mitochondrial permeability transition pore were determined by flow cytometry. The level of ATP was detected by fluorescein-luciferase. The changes of caspase-9 and caspase-3 were analyzed by Caspase Activity Assay Kit. Results HUVESs cell viability and damage of mitochondra were significantly decreased after heat stress. Compared with 43℃ heat stress group, pretreatment with propofol induced the recovery of cell viability and the ROS levels were significantly decreased in HUVEC cells (P<0.05). Meanwhile, the number of cells representing the decrease of mitochondrial membrane potential (the proportion of JC-1 monomer) was significantly decreased (P<0.05) by propofol. The average fluorescence intensity of calcein which representing the MPTP changes and intracellular ATP content was significantly increased (P<0.05). In addition, the activation of mitochondrial apoptotic pathway mediated by caspase-9/3 was also inhibited. Conclusions Propofol have anti-oxidative, anti-apoptosis and mitochondria protective effect against endothelial cell injury during heat stress.

Objective To detect the single nucleotide polymorphisms (SNPs) in the upstream promoter region of chemokine like factor (CKLF) gene and analyze their possible associations with asthma and asthma-related phenotypes.Methods Direct Sequence of the 1553bp upstream promoter region of CKLF gene was performed in 245 Chinese Han human genomic DNAs (119 asthmatics and 126 controls).The frequencies of alleles, genotypes, and haplotypes were determined and the association of these SNPs with asthmawere further analyzed.Results Fournovel SNPs,SNP88 (T>C),SNPI96 (T>C),SNP568 (C> G) ,and SNP1047 (C > G) were found in the promoter region of CKLF.The frequency of rare allele was 0.168 (SNP88C), 0.168 (SNP196C), 0.352 (SNP568G) and 0.167 (SNP1047G), respectively.Haplotypes,their frequencies and the linkage disequilibrium coefficients between SNPs were constructed.Complete linkage disequilibrium (LDs) were observed between SNP88 and SNP196,SNP88 and SNP1047, as well as SNPI96 and SNP1047 ,respectively (D1 = 1.000,r2 = 1.000).SNP568 was in partial LD with the other three SNPs (r2 = 0.366).No association between asthma and the SNPs was observed.Conclusions Four SNPs in the regulatory region of CKLF in Chinese Han population were firstly identified.Although no significant correlation with asthma was revealed, the SNP and haplotype information is useful for other disease association studies in the future.

Objective:To investigate the effect of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in modulat-ing the effects of oral squamous cell carcinoma (OSCC) invasion. Methods:Real-time polymerase chain reaction was employed to de-tect the expression of MALAT1 in samples of OSCC post-radical resection, normal oral mucosa samples, and oral squamous cell lines. MALAT1-siRNA was transfected into TSCCa human tongue squamous cell carcinoma cell lines. Cell proliferation was determined by methyl-thiazolyl-tetrazolium reduction assay. Cell migration and invasive ability were evaluated by scratch test and transwell assay. The expression of proteins that regulated invasion and apoptosis were examined using Western blot assay. Immunofluorescence assay was used to detect changes in epithelial-mesenchymal transition (EMT)-associated proteins in the cells. Tumor-bearing nude mouse models were established by subcutaneous implantation of TSCCa cells. Immunohistochemistry was used to detect up-regulation of proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2/9 (MMP-2/9). Results:MALAT1 expression was significantly higher in OSCC than in normal tissues (P<0.05). MALAT1 expression was inhibited by transfecting MALAT1-siRNA. After MALAT1 expres-sion was down-regulated in TSCCa cells, proliferation was inhibited and invasion was attenuated, showing significant differences com-pared with the cells transfected with scrambled siRNA and control cells (P<0.05). Expression of N-cadherin and MMP-2/9 were down-regulated in the cells after MALAT1 was knocked down. Tumor growth was significantly slower in the MALAT1-siRNA group than in the control groups. IHC indicated that PCNA and MMP-2/9 expression of tumor tissues were significantly inhibited in MALAT1-siR-NA group. Conclusion:MALAT1 is over-expressed in human OSCC. MALAT1 reduction can inhibit the proliferation and invasion of OSCC cells. Furthermore, MALAT1 may promote OSCC invasion and metastasis by modulating EMT.

Objective: To analyze the strengths, weaknesses, opportunities and threats of the construction on intelligent vaccination clinics in Zhejiang Province, so as to provide countermeasures for promoting the construction of intelligent vaccination clinics in Zhejiang Province. Methods: By reviewing the annual reports of Zhejiang immunization planning, survey data from Zhejiang Centers for Disease Control and Prevention and Immunization Intelligent Service System, data of human resources of immunization planning, vaccine procurement, construction progress of intelligent vaccination clinics and vaccination were collected. The relevant literature was searched to gather information on the construction standards and norms of intelligent vaccination clinics. The analysis of the strengths, weaknesses, opportunities and threats (SWOT) of the construction of intelligent vaccination clinics was conducted, and corresponding countermeasures and suggestions were proposed. Results: The National Immunization Program reported vaccine rate in Zhejiang Province is more than 99%, and standardized vaccination clinics have been popularized throughout the province. The vaccination staff are professional, and a province-wide intelligent immunization service information system has been established, providing the resources and conditions for the construction of intelligent vaccination clinics. However, there are problems such as low data quality and matching efficiency in vaccination, insufficient data interoperability and sharing, unbalanced regional capabilities in intelligent transformation, and uneven distribution of talent and resources. It is crucial to seize the opportunities presented by the development of big data and artificial intelligence, rely on the regional development of the Internet and health industry, seize the opportunity of rapid growth in demand for intelligent vaccination services and high public acceptance, accelerate the construction of intelligent vaccination clinics, and establish intelligent vaccination service standards as soon as possible. Conclusion We should seize the opportunities presented by the digital reform and development, fully utilize the existing vaccination resources and strengths, address the shortcomings, and accelerate the construction of intelligent vaccination clinics in Zhejiang Province.

Aim To design and synthesize of a new rhein derivative 1 , 8-diacetyl rhein ( 2-bromo )-ethyl ester ( rhein derivatives B ) and explore its effect on os-teosarcoma MG-63 cells and the related mechanism . Methods 1 , 8-diacetyl rhein ( 2-bromo )-ethyl ester was synthesized from rhein and its structure was identi-fied by UV, IR and NMR spectra .The purity of the synthetic product was determined by HPLC .The in vitro anti-proliferative activity of rhein and the synthetic product on osteosarcoma MG-63 cells were determined by MTT assay .Apoptosis and cell cycle distribution were detected by flow cytometry .Results The molec-ular structure of 1 , 8-diacetyl rhein ( 2-bromo )-ethyl ester was confirmed by UV , IR, 1 H NMR and 13 C NMR, and the purity was higher than 98％.The IC50 values of rhein and rhein derivatives B on osteosarcoma MG-63 cells were 110.60μmol· L-1 and 25.78μmol · L-1 , respectively .The Results of flow cytometry showed that the apoptotic rates of rhein and rhein de-rivatives B at the concentration of 80 μmol· L-1 were (6.87 ±0.53)％and (48.84 ±2.20)％, respective-ly, and the cell cycle was mainly arrested in S phase on MG-63 cells.Conclusions The anti-tumor activity of 1,8-diacetyl rhein-(2-bromo)-ethyl ester is superior to that of rhein in vitro, and the mechanisms may be associated with blocking the process of cell cycle of os-teosarcoma MG-63 cells and initiating apoptosis .

XYL1 gene,which encodes xylose reductase with dual coenzyme activity from Candida tropicalis,was transformed into Pichia pastoris X-33 by expression vector pGAPZB.The recombination strain was immobilized in Ca-alginate beads and fermentation characterization is studied using corn cob hydrolysates.Fermentation conditions were as follow:initial pH value 6.0,30℃,initial cell concentration of 20%,the Liquid volume of 28%,rotation speed 130r/min.The average xylitol yield was 37.5% on the optimum condition.This result is expected to provide a new alternative method for producing xylitol on a large scale by bioconversion.

BACKGROUNDEarly neurological deterioration (END) is a prominent issue after recanalization treatment. However, few studies have reported the characteristics of END after endovascular treatment (EVT) as so far. This study investigated the incidence, composition, and outcomes of END after intravenous recombinant tissue plasminogen activator (IV rt-PA) and EVT of acute ischemic stroke, and identified risk factors for END.

METHODSMedical records of patients who received recanalization treatment between January 1, 2014, and December 31, 2015 were reviewed. Patients were classified into IV rt-PA or EVT group according to the methods of recanalization treatment. The END was defined as an increase in the National Institutes of Health Stroke Scale (NIHSS) ≥4 or an increase in Ia of NIHSS ≥1 within 72 h after recanalization treatment. Clinical data were compared between the END and non-END subgroups within each recanalization group.

RESULTSOf the 278 patients included in the study, the incidence of END was 34.2%. The incidence rates of END were 29.8% in the IV rt-PA group and 40.2% in the EVT group. Ischemia progression (68.4%) was the main contributor to END followed by vasogenic cerebral edema (21.1%) and symptomatic intracranial hemorrhage (10.5%). Multivariate logistic regression showed that admission systolic blood pressure (SBP) ≥160 mmHg (odds ratio [OR]: 2.312, 95% confidence interval [CI]: 1.105-4.837) and large artery occlusion after IV rt-PA (OR: 3.628, 95% CI: 1.482-8.881) independently predicted END after IV rt-PA; and admission SBP ≥140 mmHg (OR: 5.183, 95% CI: 1.967-13.661), partial recanalization (OR: 4.791, 95% CI: 1.749-13.121), and nonrecanalization (OR: 5.952, 95% CI: 1.841-19.243) independently predicted END after EVT. The mortality rate and grave outcome rate at discharge of all the END patients (26.3% and 55.8%) were higher than those of all the non-END patients (1.1% and 18.6%; P < 0.01).

CONCLUSIONSEND was not an uncommon event and associated with death and grave outcome at discharge. High admission SBP and unsatisfactory recanalization of occluded arteries might predict END.