Objective Analyze the application status and existed problems of clinical biobank in China,propose possible application mechanisms for clinical biobank.Methods Through questionnaire survey and case analysis combined with relevant literature reports from home and abroad,conduct qualitative analysis to understand the application status and problems of clinical biobank.Results With the rapid development of biobank,its application rate was far from expected.The construction of clinical database lags behind in China will affect the application of samples.The lag of application mechanism will affect the opening and application of the biobank.Conclusions At the beginning of construction,the clinical biobank should take full account of what and how resources can be used,and establish tailored application mechanism.More attention should be paid to the possible benefit of biobank construction.

Objective: To investigate the prognostic impact of alterations of epidermal growth factor receptor(EGFR) and MGMT in glioblastoma. Methods: The retrospective study included 161 supratentorial glioblastomas diagnosed in the Department of Pathology, Xuanwu Hospital, Capital Medical University from 2009 to 2015. EGFR and EGFRvⅢ protein expression was detected by immunohistochemistry; EGFR amplification was detected by fluorescence in situ hybridization; MGMT promoter methylation was detected by pyrosequencing. The change of molecular genetics EGFR and MGMT and outcome were assessed statistically. Results: There were 161 patients, including 85 (52.8%) males and 76 (47.2%) females. The mean age was 53 years, and the median overall survival was 13 months. The integrated classification of glioblastoma included 16 IDH-mutant, 134 wild type, and 11 NOS. The rate of overexpression of EGFR protein was 32.9%(53/161), and that of EGFR amplification was 37.5%(18/48). There was high concordance between immunohistochemistry and FISH(85.4%, Kappa=0.475, P<0.01) and between the level of EGFR protein and EGFR amplification (P<0.01). Twelve cases showed EGFRvⅢ expression, and all also showed EGFR protein overexpression; 149 cases were EGFRv Ⅲ wild type, and EGFR protein overexpression was seen in 27.5%(41/149) of cases. There was no correlation between EGFR and EGFRv Ⅲ expression. Of all cases, 70.2%(106/151) showed MGMT promoter methylation by pyrosequencing. The changes of molecular genetics of EGFR and MGMT were not related. EGFR amplification and protein overexpression had no significant relationship with prognosis. Patients with EGFRv Ⅲ-mutant had shorter survival time than the EGFRv Ⅲ-wild type(P=0.014); patients with MGMT promoter methylation had better prognosis than without (PFS:P=0.002,OS:P=0.006),and MGMT promoter methylation was an independent predictor for overall survival (HR=0.269, 95%CI 0.124-0.583, P=0.001). Conclusions EGFR protein expression by immunohistochemistry correlates with the status of EGFR amplification. Patients with EGFRv Ⅲ-mutant tumors have poorer prognosis than that with EGFRv Ⅲ-wild type tumors. MGMT promoter methylation is closely associated with prognosis and an independent predictor for overall survival.

Objective Based on Total Quality Management (TQM) theory,our study aims to analyze the clinical Bio-Bank overall quality management implications,basic characteristics,principles,and management mechanisms,and provide theoretical basis for the clinical Bio-Bank quality construction.Methods Using theoretical and literature research methods,Bio-Bank overall quality management qualitative analysis was conducted,putting forward a framework of Bio-Bank comprehensive quality management.Results Biological sample overall quality management was defined theoretically including its connotation,concepts and basic characteristics.We also put forward an application principle and basic operation method at the application level.Conclusions Total Quality Management (TQM) is applied to the clinical Bio-Bank construction,from where,the scientific and unique management content can effectively optimize the Bio-Bank management regulation and standardization of the sample operation process in the PDCA cycle,which is critical to improve the quality of clinical Bio-Bank.

Objective: To investigate the usefulness of loss of CIC expression as the prescreening detection of 1p/19q co-deletion in the diagnosis of oligodendroglial tumors and its prognostic implication. Methods: The retrospective study included 113 oligodendroglial tumors diagnosed in the Department of Pathology, Xuanwu Hospital, Capital Medical University. Expression of CIC protein was detected by immunohistochemistry, and the 1p/19q co-deletion by fluorescence in situ hybridization in all the tumors; and the correlation of the loss of protein and 1p/19q co-deletion with prognosis was assessed. Results: The rate of negative CIC protein expression was 59.3% (67/113) in 113 oligodendroglial tumors. CIC protein expression was differentially lost in various gliomas, 85.7% (42/49) in pure oligodendrogliomas and 39.1% (25/64) in mixed oligodendroglial tumors (P<0.01). The loss of CIC protein expression showed a sensitivity of 76.1% (54/71), specificity 71.1% (27/38), false positive rate of 16.9% (11/65), and a false negative rate of 38.6% (17/44). In 63 cases integrated diagnosis as oligodendroglial tumors with mutant IDH and 1p/19q co-deletion, the loss of CIC protein expression was 81.0% (51/63); the sensitivity and specificity were increased to 81.0% (51/63) and 76.9% (20/26), and the false positive rate and false negative rate decreased to 10.5% (6/57) and 37.5% (12/32), respectively. By using Kaplan-Meier analysis, the CIC negative group showed a trend towards better outcome than the CIC positive group, but there was no statistical difference (overall survival: P=0.218; progression free survival: P=0.249). Conclusions Detection of the lost CIC protein expression can predict the chromosome 1p/19q co-deletion. In oligodendroglial tumors with IDH mutant and 1p/19q co-deletion, there is no relation between prognosis and CIC protein expression.

Objective: To investigate the diagnostic and prognostic implications of ATRX mutation and p53 mutation in patients with glioma. Methods: The clinicopathologic and molecular features of Chinese adult glioma patients, including diffuse and anaplastic astroastrocytoma with IDH mutation, oligodendroglioma and anaplastic oligodendroglioma with IDH mutation and 1p/19q co-deletion and diffuse astroastrocytoma with IDH wild type were reviewed and tested for ATRX loss expression and p53 overexpression. Results: Loss of ATRX expression was seen in 85.19% (23/27) diffuse and anaplastic astroastrocytoma with IDH mutation, higher than that of oligodendroglial tumors (0/53; P<0.01). Loss of ATRX expression was strongly linked to p53 overexpression(69.57%, 16/23). The patients who lost ATRX expression combined with normal p53 expression survived longer(P=0.013). Conclusions ATRX mutation is a molecular marker for astrocytic tumors. ATRX mutation combined with p53 mutation can predict prognosis of patients with glioma.

Objective To explore glycogen synthase kinase -3β( GSK-3β) activity and Toll-like receptor 4 ( TLR4 ) proteins expression of microglia were tested in vitro experiments, and the possible mechanism of postoperative cognitive dysfunction(POCD).Methods The cell morphology of primary culture microglia was observed by inverted microscope;microglia were identified by glial fibrillary acidic protein ( GFAP ) immunofluorescence;the best POCD modeling conditions of microglia injury induced by lipopolysaccharides( LPS) were screened ; microglia vigor was assayed by MTT ; the proteins expressions of GSK-3βand TLR4 of microglia were detected by Western blot.Results GFAP immunofluorescence showed a positive result that primary culture of rat microglia was successful;MTT result showed that the best PODC modeling conditions of microglia injury induced by LPS (100 ng/mL) was 7h; Western blot results showed that the preotein expressions of GSK-3βand TLR4 of microglial cells were up-regulated by LPS compared with the control group,and there were significantly differences (P<0.01).Conclusion PODC pathogenesis may be associated with LPS that could up-regulat the protein expression of GSK-3βand TLR4 in microglial cells.

Objective To investigate the role of the clock gene promoter methylation in aging. Methods C57BL mice of 4- (young, n=9) and 20- (old, n=10) month-old were determined the promoter methylation level of clock genes (Per1/2, Bmal1/2, Cry1/2, Clock, Npas2) in the stomach, spleen, vascular, kidney and striatum with methylation-specific polymerase chain reaction (MSP). Results The incidence of promoter methylation of Cry1, Bmal2 and Npas2 in spleen increased in old mice (P<0.05), while the promoter methylation of Per1 in stomach decreased (P<0.05), and the promoter methylation of Bmal1 in vascular increased (P<0.05). Conclusion Promoter methylation of some clock genes is involved in process of aging in a tissue-specific way.

Objective To explore the association between late-onset sporadic Parkinson' s disease (PD) and single nucleotide polymorphisms (SNPs) of Ca2+-dependent protease calpain inhibitor calpastatin (CAST) gene in a Chinese Han population.Methods 370 evaluable patients (221 male,149 female) with PD (mean age 65.2 ± 8.5 years) and 390 neurologically healthy controls (208 male,182 female) matched for gender,ethnicity,and area of residence.PD cases were identified from the PD cohort of the Chinese National Consortium on Neurodegenerative Diseases (www.chinapd.cn).A total of 24 tag-SNPs were genotyped capturing 95％ of the genetic variation across the CAST gene.Results There was no association found between any of the polymorphisms and PD in all models tested (co-dominant,dominant-effect and recessive-effect (P ＞ 0.05)).Similarly,none of the common haplotypes was associated with a risk for PD(P ＞ 0.05).Conclusion Results show no significant association between the CAST gene polymorphisms and late onset sporadic PD in the present population.

ObjectiveTo investigate the relationship between polymorphism in the PITX3 gene and hereditary susceptibility of Parkinson's disease (PD). MethodsThree PITX3 single nucleotide polymorphisms ( SNPs ),including rs2281983,rs4919621 and rs3758549 were examined in 509 late-onset PD patients ( LOPD ),290 early-onset PD(EOPD) and 494 healthy controls.Genotyping was carried out in all subjects using a ligase detection reaction( LDR).ResultsAllele and genotype frequencies did not differ between the 799 PD patients and 494 controls ( P values of genotype were 0.494,0.343,0.951 ; P values of allele were 0.369,0.297,0.823 ),between 509 LOPD patients and 494 controls ( P values of genotype were 0.522,0.350,0.630 ; P values of allele were 0.413,0.328,0.571 ),between 290 EOPD patients and 494 controls ( P values of genotype were 0.499,0.492,0.552; P values of allele were 0.321,0.301,0.931 ),and between 509 LOPD and 290 EOPD patients ( P values of genotype were 0.577,0.710,0.127 ; P values of allele were 0.346,0.472,0.077 ) for all three SNPs (rs2281983,rs4919621 and rs3758549).There were no association petween the three PITX3 SNPs and PD.ConclusionThree PITX3 SNPs do not contribute to the risk of developing PD in Chinese population.

Objective To study the expression of clock genes in Parkinson's disease(PD) and also the molecular clock machinery of PD.Methods Seventeen PD patients(nine men,eight women)and sixteen agematched controls(nine men,seven women) were investigated in this study.Bload samples were collected over a 12h span at 21:00,00:00,06:00 and 09:00.Using a real-time PCR assay,the peripheral molecular clock was examined by measuring Bmall and Bmal2 expression in total leukocytes during the dark span(from 21:00 to 09:00)in PD patients and age-matched healthy controls.Results At PD,the relative abundance of Bmall was significantly lower at 21:00,00:00 and 06:00(21:00:(22.17±4.09)vs(51.14±8.31),P=0.003,00:00:(30.30±5.45)vs(100.00±24.71),P=0.008,06:00:(19.02±3.33)vs(65.61±14.11),P=0.002).The relative Bmal2 levels in PD patients were significantly less abundant than controls at 21:00 and 00:00(21:00:(48.09±7.40)vs(84.96±9.34),P=0.005;00:00:((65.85±7.88)vs(100.00±11.78),P=0.025).Conclusion These results suggest that a peripheral molecular clock is altered in PD patients.In addition,the relative abundance of Bmall and Bmal2 was significantly lower in PD patients versus control subjects,which can provide a molecular basis to help monitor disease progression and response to investigational drugs.