To better understand the molecular characterization ,pathogenesis ,origin ,and evolutionary relationship of the first human-infecting H6N1 influenza virus ,the dataset was downloaded from the Flu and GISAID databases ,and the phy-logenetic trees were reconstructed by maximum likelihood (ML) ,maximum parsimony (MP) and Bayesian inference (BI) . Furthermore ,the evolution rate ,the most recent common ancestor (TMRCA) ,variable sites ,glycosylation sites ,cleavage sites and other some molecular features were also analyzed by bioinformatic means .The results indicated that the HA gene fell into an individual clade with other three H6N1 virus strains isolated from chicken .TMRCA was in 2002 ,and the cluster had a higher evolution rate .The molecular features revealed the HA protein had 5 N-glycosylation sites ,2 O-glycosylation sites and 2 unique variable sites .The cleavage sites had only one basic amino acid and suggested that the human-infecting H6N1 virus strain was low pathogenic .However ,because of the higher substitution rate and viral reassortment ,people should pay more attention .

Objective:To investigate the relationship between the degree of basilar artery stenosis and the short-term prognosis in patients with isolated pontine infarction.Methods:One hundred and thirty-seven patients with isolated pontine infarction within 1 month after symptom onset admitted to our hospital from April 2016 to April 2018 were consecutively included.Based on modified Rankin scale(mRS)socres, patients were divided into the good outcome group(mRS score≤2)and the poor outcome group(mRS score>2). Venous blood samples were taken for biochemical testing on admission or the next day.Baseline National Institutes of Health Stroke Scale(NIHSS)scores and demographic data were recorded and compared between the two groups.The degree of basilar artery stenosis was assessed by magnetic resonance angiography(MRA), and subjects were divided into the non-stenosis, mild stenosis, middle stenosis and severe stenosis subgroups.Results:There were 108 patients in the good outcome group and 29 in the poor outcome group.The baseline NIHSS score(2.71±0.22 vs.7.10±0.59, t=6.99, P<0.01)and total cholesterol[(4.29±0.101)mmol/L vs.(4.76±0.17)mmol/L, t=2.21, P=0.03]were lower in the good outcome group than in the poor outcome group.The proportion of patients without stenosis was higher(76 or 70.4% vs.5 or 17.2%, χ2=26.70, P<0.01)and the proportion of patients with severe stenosis were lower(4 or 3.7% vs.7 or 24.1%, P=0.002)in the good outcome group than in the poor outcome group.Binary logistics regression analysis showed that baseline NIHSS score( OR=1.658, 95% CI: 1.327-2.071, P=0.000)and degree of basilar artery stenosis( OR=2.071, 95% CI: 1.159-3.701, P=0.014)were risk factors for the short-term prognosis. Conclusions:The degree of basilar artery stenosis is a risk factor for the short-term prognosis in patients with isolated pontine infarction, and patients with severer stenosis will have a poorer prognosis.

Objective To investigate the correlation between serum uric acid level and short-term outcome of acute isolated pontine infarction. Methods From April 2016 to April 2018, consecutive patients with acute isolated pontine infarction admitted to the Department of Neurology, Kaifeng Central Hospital were enrolled. The baseline clinical data were collected. Fasting venous blood was collected on the day of admission or the morning of the next day for blood biochemical tests. The National Institutes of Health Stroke Scale (NIHSS) was used to assess the severity of neurological deficit. According to the modified Rankin Scale score at discharge or 14 dafter onset, the patients were divided into good outcome group (≤2) and poor outcome group ( > 2 ). Multivariate logistic regression analysis was used to determine the independent risk factors for short-term poor outcome. Results A total of 137 patients were enrolled in the study, 108 (78.8% ) had a good outcome, and 29 (21.2% ) had a poor outcome. The baseline NIHSS score (median [ interquartile range]: 2.5 [1.0-4.0] vs. 8.5 [5.5-10.0 ]; Z= 6.092, P< 0.001 ) and total cholesterol levels (4.290 ± 0.101 mmol/L vs. 4.763 ± 0.171 mmol/L; t=2.214, P=0.028] in the good outcome group were significantly lower than those in the poor outcome group, while serum uric acid level (329.769 ± 8.122μmol/L vs. 257.103 ± 14.290μmol/L; t=4.190, P<0.001) was significantly higher than that in the poor outcome group. Multivariate logistic regression analysis showed that high serum uric acid levels were independently associated with short-term good outcomes in patients with isolated pontine infarction (odds ratio [ OR] 0.377, 95% confidence interval [ CI] 0.203-0.702; P=0.002), while high NIHSS score (OR 1.762, 95% CI 1.375-2.258; P<0.001) and hypertension (OR 5. 353, 95% CI 1.333-21.502; P= 0.018 ) were independently associated with short-term poor outcomes. Conclusion High baseline serum uric acid levels are associated with short-term good outcomes in patients with acute isolated pontine infarction.

Objective: To investigate influences of high-voltage electrical burns on microcirculation perfusion on serosal surface of small intestine of rats and the interventional effects of pentoxifylline (PTX). Methods: Totally 180 SD rats were divided into sham injury group, simple electrical burn group, and treatment group according to the random number table, with 60 rats in each group. The electrical current was applied to the outside proximal part of left forelimb of rats and exited from the outside proximal part of right hind limb of rats. Rats in simple electrical burn group and treatment group were inflicted with high-voltage electrical burn wounds of 1cm×1cm at current entrances and exits, with the voltage regulator and experimental transformer. Rats in sham injury group were sham injured through connecting the same equipments without electricity. At 2 min post injury, rats in sham injury group and simple electrical burn group were intraperitoneally injected with 2 mL normal saline, and rats in treatment group were injected with 2 mL PTX injection (50 mg/mL). At 15 min before injury and 5 min, 1 h, 2 h, 4 h, and 8 h post injury, 10 rats in each group were selected to collect blood of heart respectively. Serum were separated from the blood to determine the level of soluble vascular cell adhesion molecule-1(sVCAM-1) with enzyme-linked immunosorbent assay method. The number of adhesional leukocyte in mesenteric venule of rats was determined with Bradford variable projection microscope system. The microcirculation perfusion on serosal surface of small intestine of rats was detected with laser Doppler perfusion imager. Data were processed with analysis of variance of factorial design and LSD test. Results: (1) At 5 min, 1 h, 2 h, 4 h, 8 h post injury, the serum content of sVCAM-1 in rats of simple electrical burn group were (8 502±1 158), (11 793±3 310), (9 960±2 146), (9 708±1 429), (7 292±1 386) ng/mL respectively, higher than that in sham injury group and treatment group [ (1 897±946), (1 882±940), (1 882±938), (1 888±946), (1 884±942) ng/mL, and (6 840±1 558), (6 742±2 465), (5 625±2 593), (2 373±1 463), (5 187±2 797) ng/mL, respectively, with P values below 0.001]. The serum content of sVCAM-1 in rats of sham injury group and treatment group at all time points post injury, except 4 h post injury of treatment group, was higher than that of the same group at 15 min before injury (with P values below 0.001). (2) At all time points post injury, the number of adhesional leukocyte in mesenteric venule of rats in simple electrical burn group was higher than that in sham injury group and treatment group (with P values below 0.001). The number of adhesional leukocyte in mesenteric venule of rats in simple electrical burn group and treatment group at all time points post injury was higher than that of the same group at 15 min before injury (with P values below 0.001). (3) At all time points post injury, the microcirculation perfusion on serosal surface of small intestine of rats in simple electrical burn group was lower than that in sham injury group and treatment group (with P values below 0.001). The microcirculation perfusion on serosal surface of small intestine of rats in simple electrical burn group and treatment group at all time points post injury was lower than that of the same group at 15 min before injury (with P values below 0.001). Conclusions High-voltage electrical burns can increase the serum content of sVCAM-1, the number of adhesional leukocyte in mesenteric venule, and reduce microcirculation perfusion on serosal surface of small intestine of rats. PTX can inhibit secretion of serum sVCAM-1, reduce the number of adhensional leukocyte in mesenteric venule to alleviate microcirculation disturbance caused by high-voltage electrical burns.

Objective To investigate the relationship between serum cathepsin S (Cat S) and cystatin C(Cys C) expression levels with carotid arterial plaque property in the patients with ischemic stroke(IS).Methods A total of 336 cases of atherosclerotic cerebral infarction were divided into the stable plaque and unstable plaque group according to the carotid arterial ultrasound.Contemporaneous 114 individuals undergoing healthy physical examination served as the control group.Fasting blood was collected from all subjects entering the groups for detecting serum Cat S and Cys C.Results The serum Cat S and Cys C levels and Cat S/Cys C ratio (Cat S/Cys C) had statistically significant difference between the two groups (P＜0.05),moreover the Cat S level in the unstable plaque group of the patients with cerebral infarction was increased compared with that in the stable plaque group,while serum Cys C level was decreased compared with that in the stable plaque group (P＜0.05).The Cat S levels were (75.34-±-15.45)pg/mL and (60.12±18.53)pg/mL and the Cys C levels were (0.73±0.62)mg/L and (0.93±0.53)mg/L,the Cat S/Cys C ratios were (103.68±2.52) and (64.64± 9.24) respectively,the difference between the two groups had statistical significance (P＜0.05).The Spearman correlation analysis showed that the serum Cat S level and Cat S/Cys C ratio had obviously positive correlation with carotid arterial unstable plaque (r=0.498,P＜0.05;r=0.753,P＜0.01);while the level of serum Cys C was negatively correlated with the unstable plaque (r=-0.213,P＜0.05).Conclusion Serum Cat S/Cys C level has a certain correlation with carotid arterial plaque property in ischemic stroke patients,which may become the serological indicators for predicting carotid arterial plaque.

Objective: To explore the influence of high-voltage electrical burns on the number of platelet aggregation, β-thromboglobulin (β-TG) and platelet factor 4 (PF-4) and the interventional effects of ulinastatin in rats with high-voltage electrical burns. Methods: A total of 240 Sprague-Dawley rats were divided into sham injury (SI) group, simple electrical burn (SEB) group, normal saline (NS) group, and ulinastatin (UTI) group according to the random number table, with 60 rats in each group. The electrical current was applied to the outside proximal part of left forelimb of rats and exited from the outside proximal part of right hind limb of rats. Rats in groups SEB, NS, and UTI were inflicted with high-voltage electrical burn wounds of 1 cm×1 cm at current entrances and exits, with the voltage regulator and experimental transformer. Rats in group SI were sham injured through connecting the same equipments without electricity. At 2 min post injury, rats in group NS were intraperitoneally injected with 2 mL/kg NS, and rats in group UTI were intraperitoneally injected with 2×10⁴ U/kg UTI of 10 g/L. At 15 min before injury and 5 min, 1 h, 2 h, 4 h, 8 h post injury, 10 rats in each group were selected to collect 5-7 mL blood of heart respectively. Blood of 0.05 mL were collected to make fresh blood smear for observing the number of platelet aggregation, and serum were separated from the remaining blood to determine content of β-TG and PF-4 with enzyme-linked immunosorbent assay. Data were processed with analysis of factorial design of variance, student-Newman-Keuls test, Kruskal-Wallis H test, Wilcoxon rank sum test, and Bonferroni correction. Results: (1) At 15 min before injury, the numbers of platelet aggregation of rats were close among groups SI, SEB, NS and UTI (5.9±1.2, 5.8±1.2, 5.9±1.3, 5.9±1.1, respectively, with P values above 0.05). At 5 min, 1 h, 2 h, 4 h, 8 h post injury, the numbers of platelet aggregation of rats in group SEB were 57.2±16.3, 59.1±16.9, 60.8±20.6, 83.6±24.9, and 83.4±30.3, respectively, obviously more than those in group SI (6.0±1.3, 6.0±1.4, 5.9±1.4, 5.7±1.1, and 5.8±1.3, respectively, with P values below 0.001); the numbers of platelet aggregation of rats in group UTI were 29.6±7.4, 31.9±10.1, 35.0±14.2, 43.0±13.6, and 35.2±11.1, respectively, obviously more than those in group NS (58.3±16.1, 63.9±18.0, 60.8±17.7, 74.2±23.0, and 82.3±21.9, respectively, with P values below 0.001). There was no significantly statistical difference in the number of platelet aggregation of rats in group SI between each two time points within the same group (with P values above 0.05), but the number of platelet aggregation of rats in the other 3 groups at each time point post injury was significantly more than that of the same group at 15 min before injury (with P values below 0.001). (2) At 2, 4, and 8 h post injury, β-TG content of serum of rats in group SEB was significantly higher than that in group SI (with Z values from -3.780 to -3.477, P values below 0.05). At 5 min and 4 h post injury, β-TG content of serum of rats in group UTI was significantly lower than that in group NS (with Z values respectively -3.477 and -3.780, P values below 0.05). There was no significantly statistical difference in β-TG content of serum of rats in group SI at all time points of the same group (χ²=0.130, P >0.05). At 2, 4, and 8 h post injury, β-TG content of serum of rats in group SEB was significantly higher than that of the same group at 15 min before injury (with Z values from -3.780 to -3.553, P values below 0.05). At 5 min, 1 h, and 4 h post injury, β-TG content of serum of rats in group NS was significantly higher than that of the same group at 15 min before injury (with Z values from -3.780 to -3.477, P values below 0.05). At 1 and 4 h post injury, β-TG content of serum of rats in group UTI was significantly higher than that of the same group at 15 min before injury (with Z values respectively -3.250 and -3.780, P values below 0.05). (3) At 2 and 8 h post injury, PF-4 content of serum of rats in group SEB was significantly higher than that in group SI (with P values below 0.05). At 2 h post injury, PF-4 content of serum of rats in group UTI was significantly higher than that in group NS (P<0.05), and at 4 and 8 h post injury, PF-4 content of serum of rats in group UTI was significantly lower than that in group NS (with P values below 0.05). At all time points, PF-4 content of serum of rats in group SI was close (with P values above 0.05). At 2 and 8 h post injury, PF-4 content of serum of rats in group SEB was significantly higher than that of the same group at 15 min before injury (with P values below 0.05). At 1, 4, and 8 h post injury, PF-4 content of serum of rats in group NS was significantly higher than that of the same group at 15 min before injury (with P values below 0.05). There were significantly statistical differences in PF-4 content of serum of rats between all time points except for 5 min post injury and 15 min before injury (with P values below 0.05). Conclusions Increasing number of platelet aggregation and abnormal secretion of β-TG and PF-4 of rats with high-voltage electrical burns can lead to microcirculation disturbance. UTI can alleviate microcirculation disturbance caused by high-voltage electrical burns by reducing the number of platelet aggregation and inhibiting secretion of β-TG and PF-4.

As confusion mounts over RNA isoforms involved in phenotypic plasticity, aberrant CpG methylation-mediated disruption of alternative splicing is increasingly recognized as a driver of intratumor heterogeneity (ITH). Protease serine 3 (PRSS3), possessing four splice variants (PRSS3-SVs; PRSS3-V1-V4), is an indispensable trypsin that shows paradoxical effects on cancer development. Here, we found that PRSS3 transcripts and their isoforms were divergently expressed in lung cancer, exhibiting opposing functions and clinical outcomes, namely, oncogenic PRSS3-V1 and PRSS3-V2 versus tumor-suppressive PRSS3-V3, by targeting different downstream genes. We identified an intragenic CpG island (iCpGI) in PRSS3. Hypermethylation of iCpGI was mediated by UHRF1/DNMT1 complex interference with the binding of myeloid zinc finger 1 (MZF1) to regulate PRSS3 transcription. The garlic-derived compound diallyl trisulfide cooperated with 5-aza-2'-deoxycytidine to exert antitumor effects in lung adenocarcinoma cells through site-specific iCpGI demethylation specifically allowing MZF1 to upregulate PRSS3-V3 expression. Epigenetic silencing of PRSS3-V3 via iCpGI methylation (iCpGIm) in BALF and tumor tissues was associated with early clinical progression in patients with lung cancer but not in those with squamous cell carcinoma or inflammatory disease. Thus, UHRF1/DNMT1-MZF1 axis-modulated site-specific iCpGIm regulates divergent expression of PRSS3-SVs, conferring nongenetic functional ITH, with implications for early detection of lung cancer and targeted therapies.