Objective:To investigate the feasibility of establishing a human renal cell carcinoma model in human peripheral blood lymphocyte-engrafted to severe combined immunodeficient(hu-PBL-SCID)mice.Methods:The biological and immunological features of mice were evaluated after intra-peritoneal injection of human peripheral blood lymphocytes(PBL)and subcutaneous implantation of human renal cell carcinoma cells(RCCCs).Results:(1)Subcutaneous tumors developed in all the mice given human PBL and RCCCs.The latency period was significantly prolonged,and the tumor size was markedly depressed,as compared with the mice given RCCCs(P

Objective To construct Nedd4 knockout bone marrow-derived macrophages(BMDM) cell line by CRISPR/Cas9 technology and to provide an effective tool for studying the function and mechanism of Nedd4 in macrophage.Methods First,three high-grade sgRNAs targeting Nedd4 gene exons were screened using the online tool before synthesized sgRNAs were inserted into the PX330 plasmid respectively.Secondly,the recombinant plasmids were transferred into BMDM cells and monoclonal cells were obtained by limiting dilution method.The protein levels of NEDD4 in monoclonal cells were detected by Western blotting.Finally,the DNA sequence of the monoclonal cells was confirmed by sequence analysis.Results One Nedd4 knockout BMDM cell line was obtained.The sequencing result showed that the Nedd4 gene had 16bp deletion mutation in this cell line.Conclusion The Nedd4 knockout BMDM macrophage cell line constructed by CRISPR/Cas9 technology will be a useful tool for studying the function and mechanism of Nedd4 in BMDM cells.

0.05). Conclusions In han-nationality people in Northern China, DRB1*04 is positively associated with Eales′ disease, suggesting that DRB1*04 may confer a major influence on Eales′ disease. Turbulence of immune function begotten by infect-agents attack may occur in the individuals with Eales′ disease due to the specific hereditary diathesis of HLA, which may cause the occurrence and development of Eales disease.

The improvement of electrophoresis for DNA fingerprinting has been carried out. The time of electrophoresis was shortened,and 1/3-1/2 of reagents were saved. The minimal amount of DNA samples for DNA analysis was 0.8?g. It is worthy to popularize and apply in practice.

Objective To investigate the effect of over-expression of ROBO4 on permeability of human renal glomerular endothelial cells (HRGECs) in high glucose medium.Methods HRGECs infected with recombinant lentiviral vector ROBO4 were cultured in high glucose or low glucose medium in vitro.The protein levels of ROBO4 and ARF6 in each group were detected by Western blotting.The endothelial permeability was measured by the effiux of fluorescein isothiocyanate-dextran (FITC-Dextran)permeated through the monolayer endothelial cells using Transwell cell model system.The cell viability after lentivirus transfection was measured by CCK8 assay.Results The transfection rate of lentiviruses in HRGECs reached 80％ 72h after,and obvious overexpression of ROBO4 protein was in transformed cells compared with the empty vector group (P＜0.05).The lentivirus-mediated ROBO4 transfection did not affect cell viability of HRGECs.Compared with the low glucose group,the expression of ROBO4 increased obviously after 12h,but declined after 24h (P＜0.05),and reached to minimun after 72h (P＜0.05).On the contrary,the expression of ARF6 increased after 12h,and the increase reached to the maximum after 72h (P＜0.05).Furthermore,the vascular permeability increased gradually after 24h,and reached to the maximum after 72h (P＜0.05) in high glucose group.Compared with the empty vector group,the over-expression of ROBO4 inhibited the expression of ARF6 significantly,and the FITC-Dextran permeability reduced obviously.Conclusion Over-expression of ROBO4 may significantly enhance the barrier functions of HRGEC in high glucose medium,and ROBO4 activation may be a potential therapeutic approach in diabetic nephropathy.

Objective To investigate the relationship between killer cell immunoglobulin-like receptors(KIR)and HLA by distribution of KIR gene in leukemia patients and their siblings who have same HLA-A/B/DR typing.Methods KIR genotypes were detected by PCR-SSP on 78 patients and their siblings who have same HLA-A/B/DR typing.Results There were 48.72%in 78 patients who had same KIR genotypes with their siblings while the 44.87%patients had different KIR genotypes with their siblings.There was no difference in frequency between patients and their siblings(P＞0.05).There were no differences in frequency among chronic myelocytic leukemia(CML),non acute lymphoblagtic leukemia(NALL)and acute lymphoblagtie leukemia(ALL)but the frequency of KIR2DS4 in CML was higher than others.Condusion The KIR gene and HLAⅠ antigen are heredity independently and relatively stable.The factor of disease has little effect on KIR gene.

G in exon 2. This results in an amino acid change from His to Asp at codon 74. Conclusion The novel allele has been confirmed as a new HLA allele and it is officially named HLA-A*0290 by WHO Nomenclature Committee for Factors of the HLA System in Oct. 2005.

Background The diseases of neuro-ophthalmology are common,which can influence the quality of patients' life seriously.The 39-item National Eye Institute Visual Function Questionnaire (NEI VFQ-39) is an instrument to assess self-reported visual impairment in studies of vision.However,until now,no studies have been performed to specifically assess the vision-related quality of life in persons with diseases of neuro-ophthalmology.Objective To evaluate vision-related quality of life for neuro-ophthalmology inpatients through the NEI VFQ-39,for the better treatment and nursing for them.Methods Ninety-seven neuro-ophthalmology inpatients were scheduled in research from June 1 to September 30,2012,and we collected all the clinical datas and the Chinese version NEI VFQ-39.The NEI VFQ-39 subscale item scores were compared among subgroups divided by monocular or binocular incidence,diseases and best corrected visual acuity(BCVA),and correlation analysis of the NEI VFQ-39 scores and BCVA for better-seeing and worse-seeing eyes was performed.Results The mean age of scheduled patients was (36.6±14.4)years,with no difference in gender.The mean composite score of VFQ-39 was 57.36(46.50,73.38),mental health subscale score was lowest [45.00 (35.00,60.00)].In the four eye diseases groups,except for eye pains,degree of dependence,periphery vision,anterior ischemic optic neuropathy (AION) group had lower scores compared with the other three groups,but there were no statistical significances among the four groups (all at P＞0.05).Except for eye pains,patients with monocular incidence had better scores than those with binocular incidence,with statistical significant significances between them(all at P＜0.05).Patients with BCVA in the better-seeing eye ＞ 20/32 had better scores than the other two groups,compared with the BCVA in the better-seeing eye ＜20/200 group,except for overall health and eye pains,cornpared with the 20/200 ≤ BCVA in the better-seeing eye ＜20/23 group,except for eye pains,social activity and color vision,and the rest subscale scores had significant differences among the three groups(all at P＜0.05).NEI VFQ-39 scores had positive correlations with the BCVA for better-seeing and worse-seeing eyes,and had strong correlations with the BCVA for better-seeing.Conclusions Neuro-ophthalmology diseases have serious influences on patients' visual function and quality of life.The quality of life has direct correlation with BCVA,so improving their visual function is helpful to their quality of life.

Objective To investigate the risk factors, diagnostic evaluation, etiology, and treatment in young patients with ischemic stroke. Methods The clinical data of young patients (age range 18 - 45 years) with ischemic stroke (within 2 weeks of stroke onset) admitted to the Stroke Center, the First Affiliated Hospital of Sun Yat-sen University from August 2008 to July 2013 were registered prospectively. Results A total of 300 patients were enroled, their age was 37. 8 ± 6. 8 years. There were 227 males (75. 7% ). The age of 84 patients (28. 0% ) was ≤35 years. The most common risk factors were smoking (43. 3% ), hypertension (38. 7% ), and hypercholesterolemia (38. 0% ). The positive result detection rates were lower in the antinuclear antibody (8/278), anticardiolipin antibody (34/250), 24-h Holter monitoring (2/60 ), and transthoracic echocardiography (38/232). According to the MRI findings, the detection rate of the white matter changes, old infarcts, single acute infarcts, and multiple acute cerebral infarcts were 40/282, 77/282, 145/282, and 137/282, respectively. The stroke subtypes of TOAST: large artery atherosclerosis (26. 7% ), smal artery occlusion (18. 0% ), cardioembolism (10. 0% ), other determined etiology (23. 0% ), and undetermined etiology (22. 3% ). Large artery atherosclerosis was mainly involved in anterior circulation (69/80) and intracranial arteries (75/80). In other definite causes, cerebral artery dissection was most common (36/69), and other causes included moyamoya disease (7/69) and infectious vasculitis (7/69), etc. The average length of hospital stay was 15. 4 d. The mean National Institutes of Health Stroke Scale score on admission was 7. 0, and the mean modified Rankin scale score at discharge was 2. 5. The complication rate during hospitalization was 9. 7% . 80. 3% and 48. 3% of patients received anti-platelet drugs and statins therapy during hospitalization.Conclusions This study used a prospective single-center method. It conducted a comprehensive analysis of risk factors, diagnostic evaluation, treatment, and etiology in current Chinese young patients with ischemic stroke. Its research data wil provide useful information for establishing a diagnostic strategy of high performance cost ratio, in-depth understanding of its pathophysiological mechanisms, and improving treatment strategies in Chinese young patients with ischemic stroke.

Objective Toinvestigatetheriskoflong-termrecurrenceofstrokeanditspredictorsin youngpatientswithischemicstroke/transientischemicattack(TIA).Methods Theclinicaldataofthe consecutive young patients (18-45 years)with ischemic stroke/TIA (within 2 weeks after onset)admitted to the department of neurology in the First Affiliated Hospital of Sun Yat-Sen University between August 2008 and July 2013 were enrolled prospectively. All patients were regularly followed up for a long time (The patients were followed up at the 1 st,6 th,and 12 th month after onset;then they were followed up once for every 6 months)in order to investigate stroke recurrence. The Kaplan-Meier curves were used to analyze the cumulative stroke recurrence rate of all patients. The last contact time for patients lost to follow was used as censored data to be enrolled in the analysis. The univariate analysis of the related risk factors for stroke recurrence using Log-rank test. Multivariate Cox proportional hazard regression was used to detect the related risk factors associated with stroke recurrence (adjusting for age and sex). The variables of the results of Log-ranktestP≤0.1wasselectedandenrolledinthemultivariateregressionanalysis.Results Atotalof 312 patients were enrolled in the analysis,including 294 with ischemic stroke and 18 with TIA. Their mean follow-up time was 34 ± 19 months. Thirty-four patients had recurrent stroke,including 23 with ischemic stroke,7 with TIA,and 4 with cerebral hemorrhage. The cumulative recurrence rates of stroke at 1 ,3 , and 6 years after onset were 6. 2%,10. 3%,and 16. 4%,respectively. The results of multivariate Cox proportional hazards regression analysis showed that hypertension (risk ratio [RR]2. 159;95% confidence interval [CI]1. 048-4. 447,P=0. 037)and cardioembolism (RR,2. 869;95%CI 1. 119-7. 357,P=0.028)weretheindependentpredictorsforstrokerecurrence.Conclusion Theoverall6-yearriskof recurrent stroke is not high in the Chinese young patients with ischemic stroke/TIA,but the risk of stroke recurrence is relatively higher in the first year. Hypertension and cardioembolism are the potential predictors of stroke recurrence;therefore,attention should be paid in clinical practice.