Proteomic characterization of plasma is critical for the development of novel pharmacodynamic bio-markers.However,the vast dynamic range renders the profiling of proteomes extremely challenging.Here,we synthesized zeolite NaY and developed a simple and rapid method to achieve comprehensive and deep profiling of the plasma proteome using the plasma protein corona formed on zeolite NaY.Specifically,zeolite NaY and plasma were co-incubated to form plasma protein corona on zeolite NaY(NaY-PPC),followed by conventional protein identification using liquid chromatography-tandem mass spectrometry.NaY was able to significantly enhance the detection of low-abundance plasma proteins,minimizing the"masking"effect caused by high-abundance proteins.The relative abundance of middle-and low-abundance proteins increased substantially from 2.54％to 54.41％,and the top 20 high-abundance proteins decreased from 83.63％to 25.77％.Notably,our method can quantify approxi-mately 4000 plasma proteins with sensitivity up to pg/mL,compared to only about 600 proteins iden-tified from untreated plasma samples.A pilot study based on plasma samples from 30 lung adenocarcinoma patients and 15 healthy subjects demonstrated that our method could successfully distinguish between healthy and disease states.In summary,this work provides an advantageous tool for the exploration of plasma proteomics and its translational applications.

Objective:To investigate the safety and efficacy of tirofiban therapy in acute cerebral infarction patients having broadened therapeutic time window.Methods:Eighty-four acute cerebral infarction patients having broadened therapeutic time window (the onset time was within 4.5-8 h), admitted to our hospital from January 2016 to May 2018, were chosen in our study. Forty-two patients (treatment group), with the informed consent of himself or his family, received emergent cerebral angiography and treated with tirofiban (the load of tirofiban was pumped via the microductal artery, and the maintenance load was continuously pumped intravenously for 48 h) right after the angiography; the other 42 patients (control group) received emergent cerebral angiography and treated with intensive antiplatelet aggregation therapy right after the angiography; intensive lipid-lowering therapy was given in both groups. The efficacy, safety and follow-up rehabilitation were compared between the two groups. According to the locations of acute cerebral infarction, patients in the treatment group were divided into anterior circulation infarction subgroup ( n=24) and posterior circulation infarction subgroup ( n=18); the efficacy and follow-up rehabilitation were compared between the two subgroups. Results:Patients from the treatment group had significantly lower National Institutes of Health Stroke Scale (NIHSS) scores 48 h, 7 d, and 10 d after treatment, and significantly higher NIHSS score difference values before and after treatment than those from control group ( P<0.05); the proportion of patents having good prognosis (modified Rankin scale [mRS] scores≤2) in the treatment group 3 months after treatment (78.57%) was significantly higher than that in the control group (52.38%), and the Barthel index in the treatment group 3 months after treatment (94.76±11.67) was significantly higher than that in the control group (85.00±15.17, P<0.05). Patients from the posterior circulation infarction subgroup had significantly lower NIHSS scores 48 h, 7 d, and 10 d after treatment, and significantly higher NIHSS score difference values before and after treatment than those from anterior circulation infarction subgroup ( P<0.05); the proportion of patents having good prognosis in the posterior circulation infarction subgroup 3 months after treatment (94.44%) was significantly higher than that in the anterior circulation infarction subgroup (66.67%, P< 0.05). There were no statistically significant differences in platelet count and coagulation tests between the treatment group and control group, and between the posterior circulation infarction subgroup and anterior circulation infarction subgroup ( P>0.05). Conclusion:Tirofiban could improve the prognoses of patients with acute cerebral infarction in broadened therapeutic time window, enjoying high effectiveness and safety, which are more obvious in the posterior circulation infarction.

Objective To investigate the association between matrix metalloproteinase (MMP)-9 gene rs20544 polymorphism and hemorrhagic transformation (HT) in patients with acute ischemic stroke (AIS). Methods Patients with AIS admitted to the Department of Neurology, TEDA Hospital from March 2016 to September 2018 were enrolled. They were divided into HT group and non-HT group depending on whether HT occurred. HT was defined as no bleeding found in the first imaging examination, and the head CT rescaning indicated a high-density lesion in the low-density area. MMP-9 gene rs20544 single nucleotide polymorphism was determined by TaqMan ? SNP genotype analysis kit. Multivariate logistic regression analysis was used to determine the independent association between rs20544 polymorphism and HT. Results A total of 204 patients with AIS were enrolled, aged 66.91 ± 9.07 years, 89 males (43.63% ), and 45 (22.06% ) developed HT. There were significant differences in atrial fibrillation, diabetes, fasting blood glucose, and triglyceride between the HT group and the non-HT group (all P<0.05). There was also a significant difference in rs2054 genotype distribution between the HT group and the non-HT group (χ2 =7.067; P=0.029 ). Multivariate logistic regression analysis showed that after adjusting atrial fibrillation, diabetes, fasting blood glucose, triglyceride, and hyperlipidemia, rs20544 CC genotype (odds ratio 2.074, 95% confidence interval 1.368-4.041) and CT genotype (odds ratio 1.571, 95% confidence interval 1.109-2.544) were the independent risk factors for HT. Conclusion MRP-9 gene rs20544 single nucleotide polymorphism is associated with increased susceptibility to HT in patients with AIS.

Objective To analyze the independent risk factors for hemorrhagic transformation (HT)after endovascular mechanical thrombectomy in patients with acute ischemic stroke (AIS).Methods From March 2015 to February 2018,patients with AIS treated with mechanical thrombectomy at the Department of Neurology,TEDA Hospital were selected.The patients with hemorrhagic infarction (HI) or parenchymal hematoma (PH) were used as the case group,and those without HT were used as the control group.The independent risk factors for HI or PH after mechanical thrombectomy in patients with AIS were determined by multivariate logistic regression analysis.Results A total of 132 patients with AIS were enrolled in the study,and 60 (45.4％) developed HT,of which 37 were HI (28.03％) and 23 were PH (17.42％).Multivariate logistic regression analysis showed that after adjusting for gender,alcohol consumption,fasting blood glucose and glycated hemoglobin,diabetes (odds ratio [OR] 3.485,95％ confidence interval[CI]l.121-6.928;P=0.019),atrial fibrillation (OR 3.962,95％ CI 1.143-7.514;P =0.007) and high fasting blood glucose (OR 3.254,95％ CI 1.107-6.549;P =0.036) were the independent risk factors for HI after mechanical thrombectomy in patients with AIS;after adjusting for gender,hyperlipidemia and glycosylated hemoglobin,diabetes (OR 3.348,95％ CI 1.120-6.709;P =0.025) and high fasting blood glucose (OR 3.172,95％ CI 1.129-7.023;P =0.014) were the independent risk factors for PH after mechanical thrombectomy in patients with AIS.Conclusion Diabetes,atrial fibrillation and high fasting blood glucose were the independent risk factors for HT after mechanical thrombectomy in patients with AIS.

Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.

Objective To study on the role of histone methylation enzyme enhancer of zeste homolog 2 (EHZ2) and vascular endothelial growth factor 165 (VEGF165) in momymoya disease. Methods The animal model of moyamoya disease was established by ear vein injection of horse serum in New Zealand rabbits. VEGF165 was over-expressed in situ by packaging lentivirus. Real-time quantitative PCR and Western Blot were used to detect the expression of VEGF165, EZH2 and H3K27me3 in the brain tissues of the animal models. Results Compared with the normal control group, the expression levels of mRNA and protein of EZH2 in the moyamoya disease model group were increased (EZH2 mRNA:P<0.01), and the level of histone H3K27me3 was increased. After overexpression of VEGF165 in the moyamoya disease model group, the expression levels of mRNA and protein of EZH2 was further increased (EZH2 mRNA: P<0.01), and the level of histone H3K27me3 was also increased. Conclusions EZH2 plays a certain role in the pathogenesis of moyamoya disease, and the expression of EZH2 is regulated by VEGF 165, which provides a theoretical basis for the study of the pathogenesis of moyamoya disease.

Objective To observe the evolution of astrocytes,GDNF,BDNF and Jak-STAT signal pathway after spinal cord ischemia-reperfusion injury in rabbits.Methods Spinal cord ischemia was induced by means of balloon occlusion of the infrarenal aorta for 22 minutes in 54 male New Zealand white rabbits.We assigned rabbits to 9 groups (n =6),one sham group,eight operation groups.The operation process in the sham group was the same as the operation group except the ischemia reperfusion of the spinal cord.At 0 h,1 h,2 h,3 h,8 h,24 h,48 h and 72 h after reperfusion,animals were sarcrificed and the spinal cord was removed for histologic,immunohistochemical study and western blotting.Results Normal neurons were decreased with the extension of reperfusion time.Levels of GFAP increased at 3 h and reached a peak at 48 h after reperfusion.GDNF was increased reaching two peaks after injury,the first peak was at 3 h,the second was at 72 h.BDNF level was increased and peaked at 24 h after reperfusion.The expression of p-STAT3 showed a biphasic pattern which peaked at 1h and 48 h.GFAP,GDNF,BDNF were rare and the level of p-STAT3 could be neglected in sham group.Conclusion Spinal cord ischemia-reperfusion injury could induce the activation of astrocytes,the expression of GDNF,BDNF and the activation of JakSTAT signal pathway.They showed different expression rules in this study.

Objective: To observe the activation of microglia and the changing rule of inflammatory cytokine as IL-6, IL-10 and nuclear factor-κB (NF-κB) in experimental rabbits after spinal cord ischemia reperfusion (SCIR) injury in order to provide theoretical basis for post-conditioning time. Methods: Rabbit SCIR injury model was established by thoracic aorta balloon occlusion. 54 New Zealand male adult white rabbits were divided into 9 groups: Sham group (the animals received balloon implantation without occlusion), SCIR-0h group (reperfusion was conducted at 0 hour of spinal cord ischemia), SCIR-1h, -2h, -3h, -8h, -24h,-48h and -72h groups. n=6 in each group. The number of normal and apoptosis neurons, the levels of Iba-1, IL-6, IL-10 and NF-κB in spinal tissue were examined and compared among different groups respectively. Results: The number of normal neuron was decreasing with the extended reperfusion time, TUNEL-positive neuron began to increasing in SCIR-8h group and the peak was reached in SCIR-24h group. The expression of Iba-1 began to elevating in SCIR-2h group and the peak was obtained in SCIR-8h group; NF-κB began to rising in SCIR-3h group and the peak was observed in SCIR-8h group; both IL-6 and IL-10 arrived the peak in SCIR-24h group. The expressions of NF-κB, IL-6 and IL-10 were positively related to Iba-1 level. Conclusion: Microglia activation had dynamic changes in experimental SCIR rabbits and the expression levels of NF-κB, IL-6 and IL-10 were positively to microglia activation; post-conditioning time at front and back to microglia activation may reduce neuron injury.

Objective To investigate the feasibility, safety and technical superiority of mechanical thrombectomy using a direct aspiration first-pass thrombectomy (ADAPT) in treatment of patients with acute cerebral artery occlusion. Methods A retrospective study was conducted on all patients with acute ischemic stroke treated with mechanical thrombectomy in our institution from January 2013 to August 2016.Patients using ADAPT or stent retriever as a first-line endovascular procedure were compared for clinical characteristics, procedural variables and clinical outcomes. The technical superiority of ADAPT was analyzed in depth. Results During observation period, a total of 91 cases were performed endovascular treatment with mechanical thrombectomy. ADAPT was designed in 46 cases as a first-line endovascular procedure and was utilized in 38 cases (82.6%;ADAPT group), while primary stent retriever thrombectomy was performed in 21 patients(stent group). There was no significant difference in baseline clinical or radiographic factors between ADAPT and stent groups. Although rates of good neurological outcome (modified Rankin Scale(mRS) score≤2) at 90 days were similar between the ADAPT and stent groups (61%(23/38) vs 48%(10/21), P=0.247), National Institute of Health Stroke Scale (NIHSS) score at seven days (6.0(2.0, 9.3) vs 9.0(5.5, 18.5),Z=-2.031,P=0.021) and full recovery rate of neurological outcome (mRS score=0, 37%(14/38) vs 10%(2/21), P=0.022) were significantly better in the ADAPT group than in the stent group. There were no significant differences in rates of embolus to new territory (21%(8/38) vs 29%(6/21), P=0.365), Thrombolysis In Cerebral Infarction (TICI) 2b/3 grade revascularization (84%(32/38) vs 81%(17/21), P=0.507) and symptomatic intracerebral hemorrhage (0%(0/38) vs 10%(2/21), P=0.123) between the ADAPT and the stent groups, but the figures were better in the ADAPT group. Conclusions Mechanical thrombectomy using ADAPT is feasible and safe compared with stent retriever, with higher full recovery rate of neurological outcome and better NIHSS score.It is a method worthy of further exploration for endovascular mechanical recanalization.

Objective: To analyze the infection characteristics of patients in acute gastroenteritis outbreaks caused by noroviruses. Methods: Between April 2014 and March 2016, the clinical data and samples were collected from the patients in acute gastroenteritis outbreaks caused by noroviruses in Beijing. Noroviruses were detected and genotyped using real time RT-PCR, and the infection characteristics of norovirus gastroenteritis were analyzed using the descriptive epidemiological method. Results: A total of 1743 clinical diagnosed cases of norovirus gastroenteritis were collected, and children under 12 years old accounted for 77.68% (1354/1743). The detection rate of noroviruses was 73.98% (509/688). The detection rates of noroviruses in fecal, swab and vomitus samples were gradually decreased (χ²=67.798, P<0.001). Among these clinical diagnosed cases, vomiting was the most common symptom (93.98%), followed by abdominal pain (40.34%), diarrhea (30.35%) and fever (27.94%). The most common symptom of patients under 6 years old was vomiting(98.14%), whereas diarrhea was most common among over 18 years old patients (68.12%). With the increase of age of the patients, the incidence of vomiting was gradually decreased (χ²=100.913, P<0.001), whereas the incidence of diarrhea was gradually increased (χ²=261.164, P<0.001). There was no statistical difference in vomiting, diarrhea and abdominal pain symptoms between patients infected with genogroup Ⅰ and Ⅱ noroviruses, and patients infected with genogroup Ⅱ (34.49%, 149/432) had the higher incidence rate of fever than that of genogroup Ⅰ (18.18%, 14/77) (χ²=7.985, P<0.01). Conclusions Noroviruses mainly infected children under 12 years old in acute gastroentertis outbreaks. The most common symptom of patients with acute gastroenteritis caused by noroviruses was vomiting, and the incidences of vomiting and diarrhea were significantly correlated with age of the patients. Patients infected with genogroup Ⅱ had the higher incidence rate of fever than genogroup Ⅰ infection.