Objective To evaluate the effect of mild hypothermia on the activity of hippocampal pro tein kinase R-like endoplasmic reticulum kinase (PERK) in a mouse model of cerebral ischemia-reperfusion (I/R).Methods One hundred and twenty male C56BL6 mice,weighing 20-30 g,aged 7 weeks,were randomly divided into 3 groups (n=40 each) using a random number table:sham operation group (group S),I/R group,and mild hypothermia group (group H).Cerebral I/R was induced by occlusion of bilateral common carotid arteries for 15 min followed by reperfusion in anesthetized mice.In group H,surface cooling was performed immediately after reperfusion,and the rectal temperature was maintained at 32-34 ℃ for 3 h.In I/R and S groups,the rectal temperature was maintained at 36.8-37.2 ℃.At 6,12,24 and 72 h of reperfusion,10 mice were sacrificed in each group,and the hippocampi were removed for determination of the number of apoptotic neurons in hippocampal CA1 region (by TUNEL),and phosphorylated PERK (p-PERK) expression (by Western blot).Results Compared with group S,the number of apoptotic neurons was significantly increased,and the expression of p-PERK was up-regulated at each time point in I/R and H groups (P＜0.05).Compared with group I/R,the number of apoptotic neurons was significantly decreased,and the expression of p-PERK was downregulated at each time point in group H (P＜0.05).Conclusion Mild hypothermia can reduce endoplasmic reticulum stress through inhibiting hippocampal PERK activity,thus attenuating cerebral injury in a mouse model of cerebral I/R.

Objective To explore the clinical value of hs-CTnT among acute myocardial infarction patients.Methods 60 acute myocardial infarction patients were treated from July 2014 to October 2016 in our hospital, set as the observation group where venous blood samples were collected on admission, 4 hours after admission, 12 hours after admission and the control group (60 health human where the venous blood samples were collected in the morning), respectively.The hs-CTnT, myohemoglobin, creatine kinase isoenzyme and tropnin T were recorded for analysis.Results On arrival of hospital, the positive rate of CK-MBmass, Myo, hs-cTnT, cTnT was 75.0%、71.7%, 90.8%, 80.0%, respectively.Hs-cTnT positive rate raised to 100.0% and CK-MBmass (83.3%), Myo (75.0%), cTnT (88.3%) after 4 hours.Besides, after hospitalization, the hs-cTnT positive rates raised up.The rates apparently increased at different times (P<0.05).The positive predictive value was 90.0% and sensibility 91.0%, which was significantly higher than other indicators (P<0.05).Conclusion The high sensitivity and specify of hs-cTnT indicator level could be one of the early diagnosis index for acute myocardial infarction.

Objective: To analyse the genomic characteristics of Novirus(NoV) GII.4 genotype JN010 strain isolated form Jinan in 2017. Methods: Seven pairs of primers were designed and used to amplify the JN010 genome. Sequence analyses, alignment and phylogenetic trees of ORF1 and ORF2 genes were performed using the software Lasergene7.1 and MEGA5.2. At the same time the major protein VP1 amino acid mutations were analyzed. Results: The 7 516 bp complete genome sequence of JN010 strain was obtained, the most mutation sites were located in P2 subdomain of VP1. Two substitutions I293N and H373N of VP1 were locate neighboring epitope A, and R297H mutation happened within epitope A and the site A that binding with histo-blood group antigens(HBGAs). The JN010 strain was GII.Pe/GII.4 genotype and genetically closest to the strains found in Osaka of Japan(GenBank accession number LC066046) and the strains in Zhongshan city of Guangdong (GenBank accession number KY407064) respectively according to ORF1 and ORF2 gene homologous and phylogenetic analysis. Conclusions The NoV GII.4 variant strain JN010 has occurred mutations in the key site of the epitope A and site A that bind with HBGAs, and maybe affect its antigenicity and interaction with HBGAs.

Objective:To evaluate the effect of mild hypothermia on inositol requiring enzyme 1-X-box binding protein 1 (IRE1-XBP1) signaling pathway in endoplasmic reticulum in cortex in a rat model of focal cerebral ischemia-reperfusion (I/R).Methods:Fifty-four clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-230 g, were divided into 3 groups ( n=18 each) using a random number table method: sham operation group (group S), cerebral I/R group (group I) and mild hypothermia group (group T). Cerebral I/R was induced by inserting a nylon thread with rounded tip into the internal carotid artery which was occluded for 2 h and then released for reperfusion.The surface cooling was started immediately after reperfusion, and the rectal temperature was maintained at 32-34 ℃ for 3 h in group T. Blood vessels were only exposed, without occlusion in group S. The neurologic deficit was assessed and scored at 24 h of reperfusion.The animals were then sacrificed and the ischemic area of the cerebral cortex was removed for examination of the ultrastructure of the cells (with a transmission electron microscope), for determination of nerve cell apoptosis (using TUNEL), for detection of the expression of IRE1 and XBP1 (by Western blot) and for determination of the expression of IRE1 and XBP1 protein mRNA (using quantitative real-time polymerase chain reaction). Results:Compared with group S, the neurologic deficit scores were significantly increased, nerve cell apoptosis in the ischemic area of the cerebral cortex was increased, the expression of IRE1, XBP1 protein and mRNA was up-regulated ( P<0.05), the neuronal nuclei was degenerated and swollen, the nuclear membrane was fragmented and defective, the chromatin was pyknotic and marginalized, and the endoplasmic reticulum was dilated and cisternal in group I and group T. Compared with group I, the neurologic deficit scores were significantly decreased, nerve cell apoptosis in the ischemic area of the cerebral cortex was decreased, the expression of IRE1, XBP1 protein and mRNA was up-regulated ( P<0.05), and the damage to the ultrastructure of nerve cells was reduced in group T. Conclusion:The mechanism by which mild hypothermia alleviates focal cerebral I/R injury is associated with further activation of neuronal IRE1-XBP1 signaling pathway and alleviation of endoplasmic reticulum stress response in rats.

Objective:To elucidate the changes of cytokine expression profiles in hand, foot and mouth disease (HFMD) patients infected with non-EV-A71 enteroviruses in Jinan city, and explore the characteristics of cytokines expression.Methods:The serum samples of acute and convalescent phases were collected from non-EV-A71 enterovirus-infected HFMD patients in Jinan from 2014 to 2017. The serum samples of healthy subjects were collected as control group. The Bio-plex liquid chip platform was used for high-throughput detection of 27 cytokines. GraphPad Prism and SPSS 22.0 were used for description and statistical analysis.Results:Twenty-two serum cytokines significantly changed in non-EV-A71 infected patients, including 11 kinds of interleukin (IL), 5 kinds of chemokines, 2 kinds of growth factors, granulocyte colony stimulating factor (G-CSF), interferon-γ, tumor necrosis factor-α (TNF-α) and granulocyte-macrophage colony stimulating factor (GM-CSF). There were 21 kinds (mean ranks 17.06-19.00 pg/ml vs 5.50-8.80 pg/ml, P < 0.05) and 20 kinds (mean ranks 16.41-19.00 pg/ml vs 5.50-9.90 pg/ml, P < 0.05) of cytokines expression in acute stage and convalescent stage respectively were higher than those in healthy control group for coxsackievirus A16 (CV-A16) infected patients, and GM-CSF expression (mean ranks 9.65 pg/ml vs 21.40 pg/ml, 9.59 pg/ml vs 21.50 pg/ml, P < 0.05) were both lower than those in healthy control group. For HFMD patients infected CV-A6, there were 19 kinds (mean ranks 11.92-13.50 pg/ml vs 5.50-6.45 pg/ml, P < 0.05) and 21 kinds (mean ranks 12.00-13.50 pg/ml vs 5.50-6.40pg/ml, P < 0.05) of cytokines expression with acute and convalescent stage respectively were higher than those in healthy control group. GM-CSF expression decreased only in acute phase (mean ranks 5.00 pg/ml vs 10.60 pg/ml, P < 0.05) compared with healthy control group. Double serum analysis showed that interleukin 6 (22.79pg/ml vs 35.88 pg/ml) and interferon-induced protein 10 (IFNγ -induced protein 10) (793.56 pg/ml vs 2 157.32 pg/ml) expression in patients with CV-A16 infection during convalescent stage were lower than that in acute stage; IL-7 (3.13 pg/ml vs 1.165 pg/ml), IL-15 (27.84 pg/ml vs 16.005 pg/ml) and regulated upon activation normal T cell expressed and secreted (RANTES) (22 605.96 pg/ml vs 7 040.90 pg/ml) expression in patients with CV-A6 infection during convalescent stage increased compared with the acute stage. Conclusions:There are extensive changes in cytokine expression profile in HFMD patients with non-EV-A71 enterovirus infection. Different pathogens infection and different clinical course of HFMD have different characteristics of cytokine expression. These findings could provide scientific data for finding indicators that are meaningful for disease progression, clinical diagnosis and immunotherapy.

Objective:To study the infection of human rhinovirus, respiratory syncytial virus and human adenovirus in Jinan from April 2018 to March 2019, and analyze epidemiological characteristics of human adenovirus.Methods:All of 1969 nasopharyngeal swabs were collected from hospitalized patients with acute respiratory tract infections in The Fourth People’s Hospital of Jinan, Qilu Children′s Hospital of Shandong University, Peoples Hospital of Zhangqiu District from April 2018 to March 2019, fluorescence quantitative PCR was used to detect the positive rate of human rhinovirus, respiratory syncytial virus and human adenovirus. Seven adenovirus positive samples were isolated and examined by sequencing, then we determined adenovirus type, constructed gene phylogenetic trees for analysis.Results:Of the 1969 samples, 242 were positive, the total positive rate was 12.30% (242/1969), the positive rate was 3.00% (59/1969) for rhinovirus, 6.30% (124/1969) for respiratory syncytial virus (RSV), and 3.86% (76/1969) for adenovirus. There was no significant difference in the detective rate of rhinovirus in different age groups (Fisher′s exact test value =8.376, P=0.720), the positive rates of RSV and adenovirus in different age groups was statistically significant ( χ2=19.28, 12.16; P=0.001, 0.016). There was a statistically significant difference in the positive rate of adenovirus between different sexes ( χ2=14.33, P<0.001), and there was no statistically significant difference in the positive rate of rhinovirus and RSV between males and females ( χ2=0.30, 2.90, P=0.862, 0.089). Comparing the positive rates of viral nucleic acid in different months, we found that the positive rate of rhinovirus, RSV and adenovirus separately reached the highest in October, December and November (8.61%, 26.50% and 8.84%). We constructed a gene phylogenetic tree after seven positive samples of adenoviruses were sequenced, by the molecular typing method we detected that seven adenovirus-positive samples were all HAdV-2 type. Conclusions:By comparing the epidemiological trends of human rhinovirus, RSV and human adenovirus in Jinan from April 2018 to March 2019 in different ages, genders, and months, providing reference basis for the early prevention and clinical treatment of acute respiratory tract infection.

Objective: To analyze the VP1 gene characteristics of Coxsackievirus A16 (CV-A16) isolated in(hand, foot and mouth disease, HFMD) patients of Jinan 2017. Methods: The samples collected from patients with HFMD in Jinan city in 2017 were analyzed and some of the CV-A16 nucleic acid positive samples were choosen with simple random sampling method and pretreated to inoculate RD cells. The whole VP1 gene sequences of CV-A16 stains which had obvious cytopathic effect in RD cells were amplified and sequenced. MEGA5.2 software was used to constructed phylogenetic tree and Lasergene software was used to analysis the homology consistency of nucleotide and amino acid of VP1 gene. Results: All the 10 CV-A16 strains isolated from Jinan were located 3 clusters belonged to B1b genotypes, with a nucleotide similarity of 94.9%~100% and deduced amino acid similarity of 99.3%~100%. The Jinan CV-A16 strains were nearest related with Shanghai strains KX871333(nucleotide similarity: 99.7%), Henan strains KM260134(99.0%), Jiangsu strains KP751580、KR138327(98.7%) and Guangdong strains MH004016(98.5%). Compared with some reference strains from our country, amino acid mutation were identified at positions 14, 59, and 145 in VP1 proteins of Jinan CV-A16 strains. Conclusions CV-A16 strains B1b genotypes were the strains prevailed in Jinan city, and 3 amino acid substitutions in VP1 proteins were happened in CV-A16 strains isolated from Jinan.