Objective : To investigate the effect of glycyrrhizin on the fibrosis of silica⁃treated mice. Methods : C57BL/6 male mice were randomly divided into control group , silicosis model group and glycyrrhizin treatment group ,with 6 mice in each group. The pathological changes of lung tissues were observed by HE and Sirius red stai⁃ ning. Lung function indexes were detected by respiratory function instrument. The content of hydroxyproline in the lung tissues was detected by corresponding kit. The mRNA levels of monocyte chemotactic protein 1 (MCP⁃1) , fibronectin (FN) and alpha⁃smooth muscle actin ( α ⁃SMΑ) were detected by real⁃time fluorescent quantitative PCR. The number of leukocytes in the bronchoalveolar lavage fluid (BALF) was counted and the secretion of transforming growth factor⁃β1 (TGF⁃ β1) in BALF was detected by ELISA. Results : HE and Sirius red staining showed that the inflammatory cells and the collagen were accumulated in the lung tissue of mice in silicosis model group. After treatment with glycyrrhizin , the accumulation of inflammatory cells and the collagen was ameliorated. Compared with the control group , pause (PAU) and enhanced pause (Penh) increased in the model group (P < 0. 05) . Glycyrrhizin treatment improved the respiratory function in mice. Furthermore , glycyrrhizin also effectively reduced the increase in the content of hydroxyproline , the expression of MCP⁃1 , FN and α ⁃SMΑ mRNA , the number of leukocytes and the secretion of TGF⁃ β1 induced by silica treatment in mice (P < 0. 05) . Conclusion Glycyrrhizin can improve the pulmonary function and alleviate the fibrosis in mice with silicosis.

OBJECTIVE: To investigate the preventive effect of rock salt aerosol on the development of silicosis in rats. METHODS: The specific pathogen free adult male SD rats were randomly divided into normal control group, rock salt control group, silicosis model group and rock salt intervention group, 18 rats in each group. Rats in the silicosis model group and the salt rock intervention group were treated with silica dust at the concentration of 2 000.0 mg/m~3 by dynamic dusting method for 3 hours daily. Rats in the rock salt control group and the rock salt intervention group inhaled the rock salt aerosols with the mass concentration of 20.0 mg/m~3 for 30 minutes daily. The normal control group was not treated with the dust or rock salt aerosol. At the time points of 14, 28 and 56 days after exposure to dust or rock salt aerosol, 6 rats were randomly selected from each group and samples were collected. The pathological change of lung was observed, the total cell count in the bronchoalveolar lavage fluid(BALF) was performed, the enzyme-linked immunosorbent assay was used to detect the change of transforming growth factor-β(TGF-β) in BALF, surfactant D(SP-D) and superoxide dismutase(SOD) in lung tissue. RESULTS: The results of hematoxylin-eosin and Masson staining showed that the inflammatory changes of lung tissue and the pulmonary interstitial fibrosis in the rock salt intervention group were less severer than that in the silicosis model group. At 14, 28, and 56 days after dust exposure, the total cell counts in BALF and SP-D levels in lung tissue of rats in silicosis model group and rock salt intervention group were higher(P<0.05), the SOD activities in lung tissue were lower(P<0.05), as well as the TGF-β levels in BALF in silicosis model group were higher(P<0.05),compared with the normal control group and rock salt control group. The total cell counts and TGF-β levels in BALF, and SP-D levels in lung tissue of rock salt intervention group were lower(P<0.05), the SOD activities in lung tissue were higher(P<0.05), compared with the silicosis model group. CONCLUSION: Rock salt aerosol intervention may delay the pathogenesis of silicosis by improving the inflammatory response, regulating oxidative stress and reducing interstitial fibrosis of lungs.

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 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 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 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 evaluate the safety and feasibility of balloon guide catheter (BGC) in recanalization of intracranial internal carotid artery occlusion.Methods Retrospective analysis was conducted in 7 patients with non-acute symptomatic intracranial internal carotid artery occlusion who underwent endovascular revascularization under the protection of BGC in our hospital from March 2016 to August 2016.The duration of occlusion ranged from 3 weeks to one year with mean time of 22 weeks.The technical aspects of using BGC were analyzed.Results Recanalization was successful in all 7 patients with an overall technical success rate of 100％,and the residual stenosis rate was＜70％.The duration time from the beginning of the first BGC filled to the end of BGC released after successfully implanted of the stent ranged from 8 minutes to 50 minutes,averaged 18.6 minutes.Among them,6 were less than 20 min;and one was 50 min,mainly because of the difficulty of micro guide wire passing through the occlusion.There was no procedural visible vascular embolization or perioperative new cerebral ischemic event,and one patient developed symptomatic cerebral hemorrhage that healed completely without any neural function loss.The patients were followed-up from one to 3 months after operation,no new cerebral ischemic event was observed,and both the improvement rate of symptoms and the patency rate were 100％ (7 of 7) in the follow-up period.Conclusion Endovascular revascularization under the protection of BGC is feasible and safe in the treatment of patients with non-acute intracranial internal carotid artery occlusion,but further investigation about patient screening,timing of surgery,recanalizing technique and postoperative management is necessary.

Objective To detect the transferred vascular endothelial growth factor (VEGF)165 gene expression in rhesus autologous bone marrow mesenchymal stem cells (MSCs), and to explore the functional viability of transgenic MSCs. Methods MSCs from rhesus bone were isolated by Ficoll, which were used to detect the phenotype. After the culturing, the expression vector pcDNA-eGFP-VEGF165 was transfected into bone marrow MSCs. Fluorescence microscope and flow cytometry were used to detect the enhanced green fluorescent protein (eGFP) expression. At the same time, the phenotype in transfected MSCs was also indentified. The VEGF165 expression level was detected by RT-PCR. Results The highly purified MSCs were collected successfully. The transfected MSCs and daughter cells showed expressions of eGFP and VEGF165, which also remained the characteristics of MSCs. Conclusion The VEGF165 gene that is transfected into MSCs can maintain characteristics of MSCs, and stably express foreign genes.

Objective To explore the influencing factors of degenerative lumbar spondylolisthesis (DLS).Methods Sixty-one DLS patients (DLS group) were enrolled in this study.The control group was formed by 61 patients with same gender and age matched.The lesion position was L4 in two groups.The correlation of intervertebral joint angle,degeneration degree,lumbosacral angle and lumbar spondylolisthesis were analyzed.Results The intervertebral joint and sagittal angle in DLS group was less than that in control group[(37.11 ± 7.52)° vs.(42.44 ± 7.61)°] (P ＜ 0.01).The lumbosacral angle between two groups had no significant difference (P ＞ 0.05).The degeneration degree between two groups had significant difference (P ＜ 0.01).Conclusion Intervertebral joint and sagittal angle and intervertebral joint degeneration degree have important roles in DLS.