Objective:To investigate the effect of Nodal protein on retinal neovascularization under hypoxia.Methods:In vivo animal experiment: 48 healthy C57BL/6J mice were randomly divided into normal group, oxygen-induced retinopathy (OIR) group, OIR+dimethyl sulfoxide (DMSO) group and OIR+SB431542 group, with 12 mice in each group. Retinal neovascularization was observed in mice at 17 days of age by retina flat mount. Counts exceeded the number of vascular endothelial nuclei in the retinal inner boundary membrane (ILM) by hematoxylin eosin staining. In vivo cell experiment: human retinal microvascular endothelial cells(hRMEC) were divided into normal group, hypoxia group, hypoxia+DMSO group and hypoxia +SB431542 group. The cell proliferation was detected by thiazolyl blue colorimetry (MTT). The effect of SB431542 on hRMEC lumen formation was detected by Matrigel three-dimensional in vitro molding method. Cell migration in hRMEC was detected by cell scratch assay. The Seahorse XFe96 Cell Energy Metabolism analyzer measured extracellular acidification rate (ECAR) of intracellular glycolysis, glycolysis reserve, and glycolysis capacity. One-way analysis of variance was used to compare groups.Results:In vivo animal experiment: compared with normal group, the neovascularization increased in OIR group ( t=41.621, P<0.001). Compared with OIR group, the number of vascular endothelial nuclei breaking through ILM in OIR+SB431542 group was significantly reduced, and the difference was statistically significant ( F=36.183, P<0.001). MTT test results showed that compared with normal group and hypoxia+SB431542 group, the cell proliferation of hypoxia group and hypoxia+DMSO group was significantly increased, and the difference was statistically significant ( F=39.316, P<0.01). The cell proliferation of hypoxia+SB431542 group was significantly lower than that of hypoxia+DMSO group, and the difference was statistically significant ( t=26.182, P<0.001). The number of intact lumen formation and migration cells in normal group, hypoxia group, hypoxia+DMSO group and hypoxia+SB431542 group were statistically significant ( F=34.513, 41.862; P<0.001, <0.01). Compared with the hypoxia+DMSO group, the number of intact lumen formation and migrating cells in the hypoxia+SB431542 group decreased significantly, and the differences were statistically significant ( t=44.723, 31.178; P<0.001,<0.01). The results of cell energy metabolism showed that compared with the hypoxia +DMSO group, the ECAR of intracellular glycolysis and glycolysis reserve in the hypoxia +SB431542 group was decreased, and the ECAR of glycolysis capacity was increased, with statistical significance ( t=26.175, 33.623, 37.276; P<0.05). Conclusion:SB431542 can inhibit the proliferation, migration and the ability to form lumens, reduce the level of glycolysis of hRMECs cells induced by hypoxia.

Objective:To observe the effect of high expression of polypyrimidine tract-binding protein-associated splicing factor (PSF) on low concentration of 4-hydroxynonenal (4-HNE) induced human retinal microvascular endothelial cells (HRMECs), and explore the possible mechanism.Methods:The HRMECs cultured in vitro were divided into 4-HNE treated group, PSF overexpression group combined with 4-HNE group (PSF+4-HNE group), PSF overexpression+ML385 treatment combined with 4-HNE group (PSF+ML385+4-HNE group), and 4-HNE induced PSF overexpression group with LY294002 pretreatment (LY294002+4-HNE+PSF group). Cell culture medium containing 10 μmmol/L 4-HNE was added into 4-HNE treatment group, PSF+4-HNE group, PSF+ML385+4-HNE group for 12 hours to stimulate oxidative stress. 1.0 μg of pcDNA-PSF eukaryotic expression plasmid were transfected into PSF+4-HNE group and PSF+ML385+4-HNE group to achieve the overexpression of PSF. Also cells were pretreated with ML385 (5 μmol/L) for 48 hours in the PSF+ML385+4-HNE group, meanwhile within the LY294002+4-HNE+PSF group, after pretreatment with LY294002, cells were treated with plasmid transfection and 4-HNE induction. Transwell detects the migration ability of PSF to HRMECs. The effect of PSF on the lumen formation of HRMECs was detected by using Matrigel in vitro three-dimensional molding method. Flow cytometer was used to detect the effect of PSF overexpression on reactive oxygen (ROS) level in HRMECs. Protein immunoblotting was used to detect the relative expression of PSF, nuclear factor E2 related factor 2 (Nrf2), heme oxygenase-1 (HO-1) protein, and phosphoserine threonine protein kinase (pAkt) protein. The comparison between the two groups was performed using a t-test. Results:The number of live cells, migrating cells, and intact lumen formation in the 4-HNE treatment group and the PSF+4-HNE group were 1.70±0.06, 0.80±0.13, 24.00±0.58, 10.00±0.67, and 725.00±5.77, 318.7±12.13, respectively. There were significant differences in the number of live cells, migrating cells, and intact lumen formation between the two groups ( t=12.311, 15.643, 17.346; P<0.001). The results of flow cytometry showed that the ROS levels in the 4-HNE treatment group, PSF+4-HNE group, and PSF+ML385+4-HNE group were 816.70±16.67, 416.70±15.44, and 783.30±17.41, respectively. There were statistically significant differences between the two groups ( t=16.311, 14.833, 18.442; P<0.001). Western blot analysis showed that the relative expression levels of pAkt, Nrf2, and HO-1 proteins in HRMECs in the 4-HNE treatment group, PSF+4-HNE group and LY294002+4-HNE+PSF group were 0.08±0.01, 0.57±0.04, 0.35±0.09, 0.17±0.03, 1.10±0.06, 0.08±0.11 and 0.80±0.14, 2.50±0.07, 0.50±0.05, respectively. Compared with the PSF+4-HNE group, the relative expression of pAkt, Nrf2, and HO-1 proteins in the LY294002+4-HNE+PSF group decreased significantly, with significant differences ( t=17.342, 16.813, 18.794; P<0.001). Conclusion:PSF upregulates the expression of HO-1 by activating the phosphatidylinositol 3 kinase/Akt pathway and inhibits cell proliferation, migration, and lumen formation induced by low concentrations of 4-HNE.

Objective:To observe the expression of miRNA in retinal tissue of mice with oxygen-induced retinopathy (OIR), and screen miRNAs related to p21 and retinal neovascularization (RNV) formation.Methods:A experimental study. Forty healthy 7-day-old C57BL/6J mice were randomly divided into normal group and OIR group, with 20 mice in each group. The oxygen induced RNV model was constructed in the OIR group, and no treatment was performed in the normal group. At the age of 17 days, the mice were killed and the RNV of mice was observed by retinal fluorescence; the nuclei of vascular endothelium that broke through the inner limiting membrane of retina were counted under light microscope. The retinal tissues were taken for miRNA chip analysis to detect the differentially expressed miRNAs between the normal group and the OIR group. The resulting differential miRNA target genes were subjected to enrichment analysis based on gene annotation (GO) and Kyoto Encyclopedia of genes and genomes (KEGG); miRNAs and pathways that may be related to p21 were screened through Targetscan, MiRanda and MicroT-CDs database alignment. Independent sample t-test was used for pairwise comparison between groups. Results:Compared with the normal group, the area of nonperfusion area, RNV and the number of vascular endothelial nuclei that broke through the inner limiting membrane of the retina in the OIR group increased significantly, differences were statistically significant ( t=18.800, 9.025; P<0.05). Compared with the normal group, there were 54 miRNAs that were statistically differentially expressed in the OIR group, of which 47 were up-regulated and 7 were down-regulated. A total of 13 miRNAs related to p21 were screened from the alignment results of the three databases with the obtained differential miRNAs. According to the difference multiples, they were miR-7218-5p, miR-322-5p, miR-224-5p, miR-335-5p, miR-329-3p, miR-362-3p, miR-532-5p, miR-20b-5p, miR-20a-5p, miR-195a-5p, miR-423-5p, miR-497a-5p, and miR-129-5p. Differential miRNA target gene enrichment analysis yielded 1 112 go entries and 50 KEGG pathways, of which 50 go entries and 13 KEGG pathways were related to p21. Conclusion:13 miRNAs related to p21 were screened out in the OIR model.

The psychological health problems of the disabled elderly in China are closely related to physical diseases. Through combing the opening of psychological care services in the pilot cities of long-term care insurance in China, it is revealed that the current pilot cities do not pay much attention to psychological nursing services, the evaluation of psychological nursing needs is absent, and the implementation regulations of psychological nursing services are not perfect. Pilot cities should strengthen psychological care services for the disabled elderly; develop an evaluation system compatible with psychological care services; improve and implement psychological care services; developing information systems that match the needs of psychological care services to ensure the linkage of data in the transition between long-term care and medical needs; and the clients and service providers should cooperate to jointly promote the sustainable development of long-term care insurance psychological care services.

Objective:To investigate the relationship between iodine nutrition level and thyroid disease.Methods:Totally 299 patients with thyroid disease who were treated at Shandong Provincial Institute for Endemic Disease Prevention and Control from 2016 to 2018 were selected as case group which was further divided into Graves' disease group (GD group, 137 patients), chronic lymphocytic thyroiditis group (HT group, 90 patients) and thyroid nodule group (72 patients). At the same time, 75 healthy people with no history of thyroid disease, normal thyroid color ultrasound and thyroid function were selected as control group. Morning urine was collected and urinary iodine was detected by arsenic cerium catalytic spectrophotometry. Fasting venous blood was extracted, and serum levels of thyroid stimulating hormone (TSH), free triiodothyronine (FT 3), free thyroxine (FT 4), thyroid globulin antibody (TgAb) and thyroid peroxidase antibody (TPOAb) were detected by electrochemiluminescence method. Results:The difference of median urinary iodine in the 4 groups was statistically significant ( H = 42.530, P < 0.05). The medians urinary iodine in GD and HT groups (326.79, 341.91 μg/L) were higher than those of thyroid nodule group and control group (235.01, 187.32 μg/L, P < 0.05). The levels of TSH, FT 3 and FT 4 in GD group were compared with those of control group, and the differences were statistically significant ( P < 0.05). The positive rates of TgAb and TPOAb in HT group were significantly higher than those in GD, thyroid nodule and control groups, and the positive rates of TgAb and TPOAb in GD group were higher than those in thyroid nodule and control groups ( P < 0.05). Conclusions:GD and HT patients have excessive iodine nutrition, and high iodine intake may lead to the occurrence of these thyroid diseases (GD and HT). Thyroid function test combined with laboratory urinary iodine test can be used to diagnose thyroid diseases simply and quickly.

Objective To analyze the relationship between blood glucose,blood lipid and body mass index (BMI) in elderly patients with hypertension.Methods Form January 2016 to December 2017,376 hypertensive patients over 65 year old in Jinan city were enrolled in this study.The sex,age,BMI,fasting blood glucose and blood lipid were investigated and tested.The prevalence of coronary heart disease and diabetes mellitus in hypertensive patients were investigated,and the relationship between BMI and blood glucose,blood lipid and liver function index were explored.Results Of 376 hypertensive patients,the detection rate of diabetes mellitus was 21.01％ (79/376),221 cases were diagnosed with coronary heart disease,with the detection rate of 58.78％.With the rise of BMI,fasting blood glucose showed an upward trend.Person correlation analysis showed that there was a significant positive correlation between BMI and fasting blood glucose(r =0.167,P =0.003).The difference between TG and TC in different BMI groups was statistically significant(FTG =12.48,P ＜ 0.001;Frc =4.83,P ＜ 0.001).The content of alanine transaminase in the overweight group and obese group was significantly higher than that in the normal body weight group,the difference was statistically significant(F =5.24,P =0.005).Conclusion For elderly hypertensive patients,it is urgent to test blood glucose and coronary heart disease screening.For people over the age of 65,obesity is still an important risk factor for hyperlipidemia and hyperglycemia.Therefore,body weight control is needed in elderly patients with hypertensive.

Objective To determine the value of using B-type natriuretic peptide (BNP) and D-dimer in preliminary recognition of cardioembolic stroke patients.Methods A mutilple-center study was conducted in Foshan Hospital of traditional Chinese Medicine (TCM) and its affiliated hospitals from July 2015 to July 2016.In the emergency departments (EDs),emergency physicians prospectively assessed consecutive adult patients with acute cardioembolic stroke and measured plasma BNP by POCT platform on admission,then followed up.Stroke neurologists evaluated patients' functional outcome at hospital discharge and also made discharge diagnosis and stroke etiologic subtypes according to the TOAST criteria.Results In this study,290 acute ischemic stroke patients met the study criteria [mean age (68.41 ± 12.06) years;53.8％ female].Of the enrolled patients,28.3％ were diagnosed with LAA at discharge,17.9％ with CE,42.8％ with SAO,11.0％ with SOE or SUE.And the mean BNP concentration was significantly higher in the CE group than that in other three subtypes (P ＜ 0.001).After adjustment for multiple clinical predictors like gender,age,coronary artery disease,atrial fibrillation and renal function,BNP and D-dimer were associated with CE [BNP OR:1.044 (95％ CI 1.025,1.064),P ＜ 0.001;D-dimer OR:1.511(95％ CI 1.020,2.238),P =0.039,respectively].Conclusion Through POCT technique in the EDs,cardioembolic stroke patients can be differentiated from other TOAST subtypes.BNP with/without D-dimer has good but different corresponding diagnostic performance in preliminary recognition of cardioembolic stroke patients.

Objective To explore the reaction time and attention bias characteristics of patients with first-episode depressive disorder.MethodsTotally 32 patients with first-episode depressive disorder (MD group) and 24 non-depression control participants(NC group) matched with MD group on age,gender and education level were enrolled in the study.The clinical symptoms were assessed by Beck Depression Inventory(BDI).All participants completed a dot-probe task to assess attentional preference for facial stimuli with varying valence (happy,sad and neutral facial expressions).ResultsThe reaction times(RTs) of MD group was longer than that of NC group in the dot-probe task ((468.6±87.7)ms,(451.7±82.5)ms,P<0.01).The four-way ANOVA revealed a significant main effect of prime duration,indicating overall shorter RTs on primes with longer duration ((476.9±88.4)ms vs (456.2±82.7)ms vs (447.7±83.9)ms,P<0.01).Compared with NC group,the scores of attention bias for sad faces were decreased in MD group ((7.43±26.4)ms vs (-4.97±19.5)ms,P<0.05).With the longer duration of presentation,the score of attention maintenance of emotional facies for MD group were increased (happy faces: (-11.0±4.8)ms,(2.2±6.9)ms,(6.1±8.5)ms;angry faces:(-1.6±7.5)ms,(6.5±8.6)ms,(14.9±6.7)ms).The adherence score of attention to happy faces were decreased ((1.8±5.6)ms,(-8.2±6.7)ms,(-8.7±7.1)ms),while the score of adherence score towards sad faces were increased ((-7.6±7.2)ms,(-2.6±8.5)ms,(1.5±6.2)ms) with increasing prime duration.ConclusionPatients with first-episode depressive disorder have slower response to emotional faces and associated with attentional bias for sad faces.With the increasing prime duration,it is more and more obvious to attentional bias in the two aspects of allocation and adherence.

Objective To summarize and analyze the epidemiological and clinical characteristics of patients with subacute thyroiditis (SAT),so as to provide a basis for prevention and treatment of the disease.Methods The epidemiological and clinical datas of 121 SAT patients,who were treated at the Shandong Provincial Institute for Endemic Disease Prevention and Control in 2015,were collected.The SAT patient's age,sex,lived in urban or rural areas,occupation and time of onset distributions were analyzed retrospectively.According to the clinical manifestations,the SAT patients were divided into types Ⅰ (mild/nodule type),Ⅱ (medium/normal),Ⅲ (severe/hyperthyroidism),and Ⅳ (delay),and clinical manifestations,auxiliary examination findings,treatment and outcome were summarized.Results Average age of the 121 cases of SAT patients was 41.36 years old,the youngest was 10 years old and the oldest was 76 years old.Most of the patients were 30-＜ 50 years old,which was accounted for 55.37％ (67/121).Most of the patients were female,which was accounted for 90.08％ (109/121).Most of the patients lived in rural areas,which was accounted for 59.50％ (72/121).Majority of the SAT patients were farmers,which was accounted for 43.80％ (53/121).Most onset of the disease was in November,accounted for 19.83％ (24/121),and the least was in June,accounted for 2.48％ (3/121),the peak of the incidence of SAT lied in autumn,and it was sporadic in the rest of the year.From the clinical classification,the mild type (61) and the medium type (46) were the largest part,a total of 107 cases,accounted for 88.43％ (107/121);while the severe type had 5 cases,accounted for 4.13％ (5/121);the delay type had 9 cases,accounted for 7.44％ (9/121).The clinical manifestations were as follows:anterior neck discomfort was 109 cases,accounted for 90.08％ (109/121);sore throat was 93 cases,accounted for 76.86％ (93/121).Auxiliary examination findings were as follows:thyroid iodine taken rate reduced in 121 cases,accounted for 100.00％ (121/121);thyroid nodule 96 cases,accounted for 79.34％ (96/ 121);total number of white blood cells increased in 52 cases,accounted for 42.98％ (52/121);neutrophil increased in 61 cases,accounted for 50.41％ (61/121);blood sedimentation significantly speed up in 49 cases,accounted for 40.50％ (49/121).All of the 121 patients were cured.Conclusions Majority of the patients are female,young aduhs and farmers.The peak of the incidence of SAT lies in autumn.The proportion of mild,medium type is the highest,we suggest this disease should be early diagnosis and treatment.

Objective:To evaluate the effect of Nutlin-3,a MDM2 antagonist,on the pyroptosis in SMMC-7721 cells.Methods: The expression of actived caspase-1(p20) and IL-1β was detected using Western blot analysis.Pyroptosis was investigated by a standard lactate dehydrogenase release assay(LDH).IL-1β content in the cell culture supernatant was quantified by ELISA.Results: Nutlin-3 up-regulated the expression level of actived caspase-1 and IL-1β in SMMC-7721 cells.Meanwhile,Nutlin-3 increased significantly the content of LDH and IL-1β in the cell culture supernatant(P<0.05).Conclusion: Nutlin-3 activated pro-caspase-1,promoted pyroptosis and IL-1β release in SMMC-7721 cells.