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

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.

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.

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.

Objective To establish a mouse model of acute liver failure induced by lipopolysaccharide /D-galac-tosamine ( LPS/D-GalN) .Methods The optimum dose of LPS/D-GalN was determined by i .p.injection of eight differ-ent doses of LPS and D-GalN into 40 female C57BL/6 mice and observation of their survival time .Then, 32 female C57BL/6 mice were i.p.injected with the optimal dose of LPS/D-GalN and sacrificed at 0, 1, 4, 8 hours after the injec-tion, 8 mice in each group.The control mice received saline injection .Hepatic changes were observed by pathology and se-rum ALT, IL-6, MCP-1 and TNF-αwere measured by biochemistry or flow cytometry .Results LPS (2.5 mg/kg) and D-GalN (0.3 g/kg) were determined as the optimal dose for the establishment of mouse model of acute liver injury .Com-pared with the control group , the hepatocellular damages were progressing in a positive correlation with the time course after LPS/D-GalN administration .The level of serum ALT was significantly increased after LPS/D-GalN administration ( P <0.001).The levels of inflammatory cytokines IL-6, MCP-1 and TNF-αwere increased and reached a peak at one hour after LPS/D-GalN administration and then decreased almost to that of the control group 8 hours later(P<0.001).Conclusions The mouse model of acute liver injury is successfully established by LPS /D-GalN administration , and provide an effective animal model for the study of pathogenic mechanisms of acute liver failure and evaluation of therapeutic drugs .

Teaching of Critical Care Medicine faces several challenging issues includingcomprehensive intensive or specialist intensive, approach of curriculum, tralning of team working ability, building of teaching platform and teaching staff. Critical care medical education requires the concept of viewing the discipline as a whole. Under its guidance and with the opportunity of critical care medical subspecialties building, critical care medical education should focus on bothcompre-hensive critical care and specialist critical care, and have rational planning of Critical Care Medicine course. Through the construction and integration of ICU, we should create a comprehen-sive clinical practice platform of critical care medicine to carry out clinical practice and team work tralning. Meanwhile, construction of quality critical care medicine faculty should be based on its pro-fessional features.

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.