Objective Objective To discuss the teaching pattern of experimental classes,and to probe the deep development of the functional subject about experimental teaching and the approach of cultivating talents.Methods Student volunteers participated in a series of activities in the summer vocation including experimental design,implementation,results reports and discussion.Results The reflection of students for the learning pattern were as follows: providing a good opportunity of learning(100%);helpful for improving the abilities of finding and dealing with problems(87.50%);different from the past physiological labs including representing special subject characteristics(93.75%),more precise form(68.75%),excellent interaction between teachers and students(75%);improving the manipulation abilities(93.75%).Deficiencies: the experimental conditions were not good enough(68.75);the teachers should give more guidance in theory and techniques(50%).Conclusions The is a meaningful educational form.Under the new experimental teaching pattern,students could use basic medical knowledge and experimental techniques to design and accomplish experiments voluntarily.This pattern is propitious for cultivating integrated capabilities of students,helping them to learn the basic approach and technique means.And it will have much developing space in experimental education in future.

Objective:To construct a p53-fused dual luciferase reporter and to test whether this reporter can mimic wild-type p53 activities in a high-throughput screen.Methods:A restriction endonuclease site was added to each terminus and the stop codon of the wild-type full-length p53 open reading frame (ORF) was removed by PCR. A restriction endonuclease site was added to each terminus and the start codon of the ifrelfy luciferase ORF was removed by PCR. The two modified ORFs were inserted upstream of the IRES-induced renilla luciferase ORF in a CMV-derived vector. hTe p53 fusion protein was expressed in cells to test its MDM2-mediated degradation, subcellular localization, and induction of p53-responsive promoter.
Results:hTe p53-fused dual luciferase reporter was successfully constructed. Atfer transfection into the host cells, the reporter expressing the p53 fusion protein that was degraded by oncoprotein MDM2, was mainly located inside the nucleus, and induced the p53-responsive promoter, respectively.
Conclusion:hTe p53-fused dual luciferase reporter (p53FL/IRES/RL) can identify modulators of P53 protein level in a high-throughput screen of genetic or chemical libraries.

OBJECTIVE: To determine the inhibitory effect of miRNA-381 on renal carcinoma invasion and to explore the underlying mechanisms.
 METHODS: After up-regulation of miRNA-381, the inhibitory effect of miR-381 on cell invasion was investigated. We screened the target genes of miRNA-381 in a database (starBase) through combination of five programs including targetscan, picTar, RNA22, PITA and miRanda. Then, the predicted targeting genes were verified by the dual luciferase reporter assay. We also examined the expression of miRNA-381 and its target genes in renal cancer cells and tissues.
 RESULTS: Transfection and up-regulation of miRNA-381 resulted in a significant decrease in trans-membrane cell numbers and the ability of renal cell invasion. Bioinformatics analysis showed that CREB binding protein (CBP), β-catenin and lymphoid enhancer binding factor-1 (LEF-1) were the potential targets of miRNA-381. In the luciferase reporter gene system, co-transfection of miRNA-381 with the 3'UTR of wild-type target gene led to a significant decrease in luciferase activity. The expression of miRNA-381 was decreased in various renal cancer cells, and it was particularly lower in highly metastatic cell lines (786-OHM). On the contrary, the expression levels of miRNA-381 target genes (CBP, β-catenin and LEF-1) were significantly increased in cells and tissues.
 CONCLUSION MiRNA-381 can inhibit cell invasion in renal cancer by block the function of CBP, β-catenin and LEF-1.
3' Untranslated Regions ; CREB-Binding Protein ; metabolism ; Carcinoma, Renal Cell ; pathology ; Cell Line, Tumor ; Computational Biology ; Gene Expression Regulation, Neoplastic ; Humans ; Kidney Neoplasms ; pathology ; Lymphoid Enhancer-Binding Factor 1 ; metabolism ; MicroRNAs ; genetics ; Neoplasm Invasiveness ; genetics ; Transfection ; Up-Regulation ; beta Catenin ; metabolism

To analyze types of urinary calculi and patients' clinical characteristics, and to explore the strategies for prevention and treatment of urinary calculi.
 Methods: A total of 1 849 patients with urinary calculi were treated in the Department of Urology, the Third Xiangya Hospital of Central South University. The components were analyzed by infrared spectroscopy. The relationship between stone composition and clinical parameters was analyzed according to the clinical characteristics of the patients.
 Results: The proportion of calcium oxalate stone or uric acid stone in male (84.1% or 7.7%) was higher than that in female (78.4% or 4.2%). The older patients were more likely to be diagnosed as uric acid stone. The proportions of uric acid stone in patients <18 years old, 18-<41 years old, 41-<66 years old, and ≥66 years old were 0.0%, 1.6%, 6.6%, and 12.4%, respectively. There was no significant difference in the proportion of stones in patients with different BMI. There were no significant difference in the stone composition between the patients with or without urinary tract infection, hypertension or diabetes. The proportion of uric acid stones in patients with acidic urine was higher than the other types. The proportion of uric acid stones in patients with elevated creatinine (12.1%) was higher than that in the patients with normal creatinine (4.5%).
 Conclusion: Elderly patients, or patients with high uric acid and renal insufficiency are more prone to uric acid stones. Regulation of urinary pH may be an important strategy for preventing and treating urinary calculi in Hunan Province.
Adolescent ; Adult ; Age Factors ; Aged ; Body Mass Index ; Calcium Oxalate ; analysis ; Creatinine ; urine ; Female ; Humans ; Hydrogen-Ion Concentration ; Kidney Calculi ; chemistry ; therapy ; Male ; Middle Aged ; Sex Factors ; Spectrophotometry, Infrared ; Uric Acid ; analysis ; Urinary Calculi ; chemistry ; therapy ; urine