Objective To observe the morphological changes of mice cortex neuron cultured in vitro under different temperature, and the expression of skeleton protein (?-tubulin) in the neuron, and to study the relationship between ?-tubulin and heat shock protein 70 (HSP70). Methods The cerebral cortex neuron of embryonic mice was cultured in vitro. The cultured neuron was put in different temperature 7 days later. To observe the morphological changes of the neuron using optical microscope and the changes of the expression of ?-tubulin and HSP70 under different temperature using laser scanning confocal images. Results Optical microscopy indicated that drifting cells increased, and neural network became sparse in 38℃; some cells necrosed in 39℃; most cells necrosed, cell broke to pieces, axons drifted or disappeared in 42℃. Results of laser scanning confocal images indicated that after hyperthermia the fluorescence intensity of ?-tubulin was lower than that of controls, and the fluorescence intensity declined as the temperature elevated. The fluorescence intensity of HSP70 showed a bell-shape distribution curve, i.e. the highest value emerged at 39℃, whereas the lower values appeared at 37℃ and 42℃. Conclusion Heat stress leaded to the morphological changes of neuron. The disordered of skeleton protein may be responsible for the changes and HSP70 may take part in the process.

Objective To explore the effects of Zhu Ling decoction and Alisma orientalis on the osteopotin mRNA expression in the experimental rat model of renal stone. Methods The experimental rat model of renal stone was set up using glyoxylic acid. Osteopontin mRNA expression in the renal tissues in the experimental rat model of renal stone was detected by RT-PCR. Results Glyoxylic acid obviously increased osteopontin mRNA expression in the renal tissues. Zhu Ling decoction and Alisma orientalis remarkably decreased the expression of osteopontin mRNA in the renal tissues. Conclusion Zhu Ling decoction and Alisma orientalis could decrease the osteopotin mRNA expression in the renal tissues in the experimental rat model of renal stone. The results were helpful for prevention and treatment of renal stone.

Objective To provide data evidence for brucellosis prevention by a retrospective analysis of brucellosis epidemiologic feature in Ji'nan Infectious Disease Hospital from 2010-2014.Method The brucellosis patients with confinned diagnosis from 2010 to 2014 admitted to Ji'nan Infectious Disease Hospital were included in this study,and their epidemic data were statistically analyzed,including time,age and occupation distributions.Results There were 1 010 brucellosis patients in total in Ji'nan Infectious Disease Hospital from 2010 to 2014,and 12 cases,69 cases,134 cases,274 cases and 521 cases,respectively,in each year,which showed rapid rising trend.There were two peak times,the first was from April to August,the incidence was 9.1％ (92/1 010),12.2％(123/1010),13.4％ (135/1010),11.1％ (112/1010) and 10.7％ (108/1010),respectively,in each month;the second was January,the incidence was 9.4％ (95/1010).The diagnosed rate within 1 week of onset only accounted for 13.4％ (135/1010),and 72.4％ (731/1010) patients were diagnosed for more than two weeks after the onset.The male incidence of brucellosis was higher than the female,and the sex ratio was 2.36:1.00.In terms of age distribution,40-49 and 50-59 were both peak ages,with incidence of 29.3％ (296/1 010) and 25.0％ (252/1010),respectively.The professional was given priority to farmers and herdsmen (85.6％,865/1 010),and 96.4％ (974/1010)of the patients had a clear exposure history,mainly related to sheep,cattle,pigs and other livestock,also related to pets and milk.Conclusions The brucellosis patients that received in Ji'nan Infectious Disease Hospital are increased significantly in the recent 5 years,and their transmission is closely associated with livestock breeding and processing.Most patients have failed timely diagnosis in the early days,which should catch enough of our attention.

Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.

Objective To compare distribution difference in risk factors of patients with first-ever ischemic stroke (IS) of different age and gender.Methods A total of 1027 patients admitted to the neurological department in Shanghai Renji Hospital with first-ever IS were recruited and divided into young adult group ( ＜ 50 years old),middle-aged group (50-80 years old),and very old group ( ＞ 80 years old)according to their ages.Risk factor analysis included history of smoking,high alcohol consumption,hypertension (HT),diabetes mellitus (DM),heart diseases,atrial fibrillation (AF) and family history of cardiovascular diseases.Results Female patients were globally older than male patients (71.1 vs 65.7,P ＜ 0.001 ) at the first attack of IS and having higher prevalence of DM (26.8％ vs 19.2％,P =0.004 ),heart diseases (28.8％ vs 19.2％,P＜0.001) and AF (7.6％ vs 3.9％,P=0.009).However,female patients were less likely to drink heavily ( 1.0％ vs 31.6％,P ＜0.001 ) or smoke (4.4％ vs 59.9％,P ＜0.001 ) than the male patients.The rates of smoking and heavy drinking in young adult group were higher than that in other two groups.Patients in very old group had higher prevalence of heart diseases and AF but lower proportion of positive family cardiovascular diseases history than patients in other two groups.HT and DM were equally frequent among three groups.In young adult group,female patients were more likely to have heart diseases and family history of heart diseases (P =0.015 and P =0.048).In middle-old group,HT,DM,heart disease and AF were more common in women than in men (P =0.021,P =0.004,P =0.001 and P =0.039).Conclusion There are differences in risk factor distribution in patients with first-ever IS of different age and gender.Therefore,screening and health education should be performed in allusion to different risk factors.

ObjectiveTo explore the damage of blood vessels in carbon monoxide poisoning patients by hyperbaric oxygen, especially for damage of vein, to understand the factors aggravating inflammatory response, thus work out the homologous protective measures.MethodsFrom January 2006 to January 2008, 80 patients of carbon monoxide poisoning were selected as the observation group. Another 80 patients with other diseases were set as the control group. Two groups both accepted hyperbaric oxygen treatment. The time of angioplerosis was compared between the two groups, besides, pain reaction of the observation group was also observed.ResultsThe time of angioplerosis was significantly different between the two groups, pain sensibility intensified in the observation group compared with that before treatment.ConclusionsHyperbaric therapy is closely related with vascular inflammatory reaction of carbon monoxide poisoning patients. Protective measures should be established in order to prevent further damage of vessel, relieve pain of patients and ensure smooth progress of the treatment.

Objective To respectively analyze the patterns and possible predictors of recurrent strokes among patients with initial ischemic stroke.Methods Three hundred and sixty-one patients with recurrent strokes (acute ischemic stroke or intracerebral hemorrhage) after initial ischemic strokes were collected from Jan 2004 to Dec 2009.The data about conventional risk factors such as smoking,heavy alcohol drinking,hypertension,diabetes,hyperlipidemia,heart diseases,head trauma,migraine,family history of cardiovascular disease,and the use of preventive medications were collected and analyzed among patients with different types of recurrent strokes.Results Patients (n =361) were divided into ischemic stroke group (n =321) and hemorrhagic stroke group (n =40) according to the recurrent stroke type.The ischemic stroke group was further divided into the anterior circulation stroke subgroup (n =234),the posterior circulation stroke subgroup (n =75) and watershed cerebral infarction or multiple infarction subgroup (n =12).Multivariate logistic regression analysis revealed that older age at initial stroke onset (OR =1.036,95 ％ CI 1.006-1.067,P =0.02) and hyperlipidemia (OR =2.253,95 ％ CI 1.092-4.647,P =0.028) were both the independent risk factors for the recurrent ischemic stroke.Comparing the subgroups,multivariate logistic regression analysis showed that atrial fibrillation (OR =4.217,95％ CI 1.489-11.942,P =0.007) was the independent risk factor for the recurrent ischemic stroke in the posterior circulation territory.Conclusion Aging and hyperlipidemia are possible predictors of recurrent ischemic stroke after the initial ischemic stroke which would be useful for individualized secondary prevention of stroke.

Objective:To explore the expression of long non-coding RNA LINC00630 in bladder cancer cell lines, and to explore the effect of interference with its expression in vitro on the proliferation and migration of bladder cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of LINC00630 in bladder cancer cell lines 5637, BIU-87, T24, J82 and normal bladder epithelial cell line SV-HUC-1. The bladder cancer cell line with the highest LINC00630 expression was selected for follow-up experiments, then the cell line infected with the control lentivirus was used as the control group, and the cell line infected with the lentivirus that could interfere with the expression of LINC00630 was used as the experimental group. qRT-PCR was used to detect the expression of LINC00630 in the two groups of cells. MTS method and cell scratch test were used to detect the proliferation and migration abilities of cells in the two groups. qRT-PCR was used to detect the expression of neuregulin 1 (NRG1) mRNA in the two groups of cells, and Western blot was used to detect the expressions of NRG1 protein, cell proliferation-related proteins (cyclin D3 and CDK2) and cell migration-related proteins (Vimentin and N-cadherin) in the two groups of cells.Results:Compared with SV-HUC-1 cells (1.05±0.17), the expression of LINC00630 was significantly increased in all bladder cancer cell lines (all P < 0.01), and the expression was highest in J82 cells (relative expression 5.83±0.42). Compared with J82 cells of the control group, the expression of LINC00630 in J82 cells of the experimental group decreased (0.18±0.02 vs. 1.00±0.05, t=14.36, P < 0.01); from day 2 of transfection, the cell proliferation activity of the experimental group was lower than that of the control group (all P < 0.05). The cell scratch closure rate of the experimental group was lower than that of the control group [(27.4±7.1)% vs. (66.0±5.4)%, t = 4.31, P < 0.01]. Therelative expression of NRG1 mRNA in the experimental group was lower than that in the control group (0.34±0.03 vs. 1.07±0.24, t = 2.99, P < 0.05). Compared with the control group, the expressions of NRG1 protein, cell proliferation-related proteins and cell migration-related proteins in the experimental group were reduced. Conclusions:LINC00630 is up-regulated in bladder cancer cell lines, and interference with LINC00630 may inhibit the proliferation and migration of J82 cells by down-regulating the expression of NRG1 gene. LINC00630 may be a new molecular target for the treatment of bladder cancer.

BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) have made certain curative effect on hepatic failure, but little is reported on the effect of hepatic failure patient's serum microenvironment on UC-MSCs differentiation ability. OBJECTIVE: To investigate the effect of hepatic failure patient's serum on the differentiation of umbilical cord mesenchymal stem cells into hepatic cells. METHODS: UC-MSCs were isolated by tissue adherent method and the cell morphology and phenotype identified by microscope and flow cytometry. Alpha-MEM media with serum before/after plasmapheresis were used to culture the hirdgeneration of UC-MSCs, and regular fetal bovine serum culture medium acted as control group. Inverted microscope was used to observe the cell morphology in three groups. Immunohistochemical method was used to measure expression level of alpha fetoprotein, albumin and cytokeratin 18. RESULTS AND CONCLUSION: A great amount of high-purity UC-MSCs could be obtained using tissue adherent method, which highly expressed CD44, CD73, CD90, CD10, but did not express CD45. Hepatic failure patient's serum could change the morphology of UC-MSCs and induce UC-MSCs to express alpha fetoprotein, albumin and cytokeratin 18. The positive expression of alpha fetoprotein, albumin and cytokeratin 18 was significantly increased after plasmapheresis (P < 0.05). To conclude, hepatic failure patient's serum after plasmapheresis exert more benefits to induce UC-MSCs to differentiate into hepatic cells than that before plasmapheresis.

Objective To evaluate the association between 8 single nucleotide polymorphisms (SNPs)of vascular endothelial growth factor (VEGF ) gene and the risk of Alzheimer’s disease (AD ) in Shaanxi Han population.Methods We examined the potential association between AD and 8 SNPs of VEGF gene using the MassARRAY system.The participants enrolled in this study included 214 patients with AD and 249 healthy controls from Shaanxi Han population.SPSS16.0 and Haploview 4.2 were employed to analyze differences in genotypes, alleles and haplotypes between the two groups.Results The results showed that rs3025039 (3’UTR)were significantly associated with AD (P<0.05).Greater frequency of rs3025039 T allele (P=0.008,OR=1.527,95%CI=1.116-2.088)was found in AD subjects.Furthermore,strong linkage disequilibrium (LD)was observed in 2 locks (block1:rs699947-rs1570360-rs2010963;block 2:rs3024997-rs3024998-rs3025006)(D’>0.9).There were no significant haplotypes in block 1 and block 2 (P=0.034)found between the patients and controls.Conclusion These findings point to the role for VEGF gene polymorphisms (rs3025039)in AD of a Shaanxi Han population. Individuals with T allele of rs3025039 may be at a higher risk for AD.