Objective To investigate the effect of tumor necrosis factor receptor 1 (TNFR1) in angiogenesis and neurogenesis during cerebral ischemia in mice.Methods Twenty-four wild-type and 24 TNFR1 knockout mice were randomly divided into either a sham operation group or a focal cerebral ischemia group (n =12 in each group).5-bromodeoxyuridine (BrdU) was injected intraperitoneally at day 3 after cerebral ischemia and sham operation.At day 7 and 28 after cerebral ischemia,the double-label immunofluorescence staining of glucose transporter-1(Glut-1)/BrdU was used to evaluate the angiogenesis surrounding the areas of infarction.A labeled BrdU was used to detect the neural stem cell proliferation in the subventricular zone.Double-labeled doublecortin (DCX)/BrdU and neuronal nuclei antigen (NeuN)/BrdU were used to detect the migration and survival of neural stem cells,respectively.Results Under the normal condition,there was no significant difference in angiogenesis and the number of BrdU-positive cells in the subependymal zone (SVZ) between the wild-type (418.000 ± 28.404) and TNFR1 knockout (528.000 ± 60.597) mice (t =-1.644,P =0.131).At day 7 after cerebral ischemia,the number of Glut-1/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wild-type mice (14.833 ± 2.182 vs.27.5 ± 4.209) (t =2.672,P =0.023),and the number of DCX/B3rdU-positive cells was also significantly less than that in the wild-type mice (163.000 ± 11.106 vs.257.168 ± 12.213) (t =5.705,P =0.000).At day 28 after cerebral ischemia,the number of NeuN/BrdU-positive cells in the TNFR1 knockout mice was significantly less than that in the wildtype mice (6.000 ± 0.577 vs.11.000± 1.571) (t=2.988,P=0.014).Conclusions TNFR1 may play a promoting role in the neurovascular reggeneration in late cerebral ischemia.

Objective In order to further understand the bioactivity of sphingosine kinase, high efficient replication defective recombinant adenovirus carrying the wild type SPK gene (SPK WT ) and SPK mutant gene (SPK DN ) were constructed respectively. Methods The SPK genes of interest were subcloned into a shuttle vector pshuttle cmv respectively. Pshuttle cmv SPK WT and Pshuttle cmv SPK DN were linearized by PmeI and transformed respectively into the competent cells of E. coli BJ AD 1 cells which contain adenoviral backbone plasmid pAdEasy 1. The linearized recombinant plasmids were transfected into adenovirus 293 packaging cells respectively,and then recombinant adenoviruses were harvested. We performed the PCR, Western blot and enzyme assays to identify the recombinant adenoviruses. Results The recombinant adenoviruses containing the interest gene and being able to infect ECV 304 cells were obtained. Overexpression of wild type SPK gene in ECV 304 cells increased the endogenous SPK activity, whereas overexpression of mutant SPK (SPK DN ) inhibited intracellular SPK activity. Conclusion Recombinant adenoviral vectors can mediate interesting gene expression in cells.

Objective To investigate the correlation between plasma cystatin C (CysC) level and carotid atherosclerotic plaque in patients with ischemic stroke.Methods The clinical data in patients with acute ischemic stroke were analyzed retrospectively.According to the results of carotid artery ultrasound,the patients were divided into either a non-plaque group or a plaque group.Then the plaque group was redivided into a stable plaque subgroup and a vulnerable plaque subgroup.Multivariate logistic regression analysis and Pearson correlation analysis were used to explore the risk factors for carotid atherosclerotic plaque.Results A total of 226 patients with acute ischemic stroke were enrolled,172 of them had carotid plaque,and 54 had no plaque.Of the patients with carotid plaque,94 were stable plaque and 78 were vulnerable plaque.The age (71.82 ± 9.94 years vs.60.74 ± 13.81 years; t =6.160,P =0.014),proportion of patients with ischemic heart disease (11.6％ vs.1.9％; x2=6.169,P=0.020),systolic blood pressure (148.770± 21.007 mm Hg vs.142.240 ± 19.404 mm Hg; t =2.029,t =0.044),plasma CysC concentration (1.046 ± 0.438 mg/L vs.0.860 ±0.214 mg/L; t =3.006,P =0.003),and carotid IMT (1.122 ±0.278 mm vs.0.878 ±0.250 mm; t =5.762,P=0.000) in the plaque group were significantly higher than those in the non-plaque group.Multivariate logistic regression analysis showed that the age (odds ratio [OR] 1.079,95％ confidence interval [CI] 1.044-1.116; P=0.000) and IMT (OR 31.450,95％ CI 6.233-158.692; P=0.000) was the independent risk factor for carotid plaque,while there was no significant independent correlation between the plasma CysC level and carotid plaque (P =0.217).Only IMT in the stable plaque subgroup was significantly higher than the vulnerable plaque group (1.176 ±0.285 mm vs.1.058 ±0.258 mm; t =-2.824,P =0.005),and it was the independent protective factor for the carotid plaque stability (OR 0.195,95％ CI 0.059-0.064; P =0.007).Pearson correlation analysis showed that the plasma CysC level was positively correlated with the age (r =0.375,P =0.000) and serum creatinine level (r =0.462,P =0.000),but it was not significantly correlated with carotid IMT (r =0.075,P =0.264).Conclusions In patients with ischemic stroke,no correlations were found between the plasma CysC level and carotid atherosclerotic plaque,plaque stability,and IMT.

Objective To assess sleep quality and daytime sleepiness in patients on maintenance high flux hemodialysis, and discussed the associated factors. Methods A total of 112 high flux hemodialysis patients and 53 normal subjects were estimated by Pittsburgh sleep quality index (PSQI) and Epworth Sleep Scale (ESS) to assess the sleep quality and day time sleepiness. Global score of these questionnaires were analyzed. Seven components' scores and 9 reasons for sleep disturbances were compared between "good" (global PSQI ≤5) and "bad" (global PSQI＞5) sleepers. Sleep quality was compared among different shifts of hemodialysis. The impact of clinical factors on sleep quality were analyzed by multivariate linear regression and logistic regression. Results Compared with control group, hemodialysis group had a higher PSQI (7.02±4.94 vs 3.28±2.79, P＜0.05) and a lower ESS score [3(0-6) vs 8(4.25-11.75), P＜0.05] . 58% patients were "bad" sleepers and sleep latency was longer (30 min vs 15 min, P＜0.05). Insomnia was the main problem. Patients on morning shift, afternoon shift and night shift had similar subjective sleep quality. Age (OR=1.75, P=0.003), dialysis vintage (OR=1.26, P=0.008),hemoglobin (OR=0.64, P=0.008), calcium phosphate product (OR=1.60, P=0.02) were significantly related to sleep quality score. Conclusions Sleep disturbance is common in hemodialysis patients. Older age, longer dialysis vintage, anemia and higher calcium phosphate product are risk factors for poor sleep quality.

Objective:To investigate the present situation of anticipatory grief of main caregivers of elderly patients with acetabular fractures and to explore its influencing factors.Methods:Using the convenient sampling method, a total of 168 main caregivers of elderly patients with acetabular fractures who underwent surgical treatment in the First People's Hospital of Fuyang District in Hangzhou and were discharged from the hospital from January 2018 to December 2019 were selected as research objects. General information questionnaire, Family Disease Burden Scale and Anticipatory Grief Scale were used to investigate.Results:The score of Anticipatory Grief Scale of primary caregivers for elderly patients with acetabular fractures was (69.61±6.35) . The results of multiple regression analysis showed that the age of the main caregivers of elderly patients with acetabular fractures, the relationship with the patients, the time of care and the degree of family disease burden were the main influencing factors of anticipatory grief ( P<0.05) . Conclusions:The main caregivers of elderly patients with acetabular fractures have a moderate level of anticipatory grief. While paying attention to the psychological problems of patients, medical staff should also include caregivers in the object category of psychological care and actively explore effective nursing measures to promote the physical and mental health of caregivers.