Objective To establish quantitative method for detection of methylation level of DLC-1 promoter with HRM technology to analyze its association with pathological parameters in prostate cancer.Methods 89 prostate cancer tissue samples and 10 matched normal tissue samples were enrolled into this study.Prostate cancer cells were obtained by LCM.DNA was extracted and modified for methylation determination.CpGenome Universal Methylated DNA was chosen as 100% methylation sample.Then the calibrators representative of 100%,80%,50%,30%,10% and 0% methylation levels were prepared with dilution in a DNA sample of peripheral blood from healthy subjects(100% non-methylation).The DLC-1 methylation levels in prostate cancer tissue samples were detected with HRM.The associations of methylated level with age of patients,PSA value,TNM stage were investigated respectively.ResultsThe melting curves representing 100%,80%,50%,30%,10% and 0% methylation levels were aligned from right to left.The methylation levels of 10 adjacent normal samples and 35 prostate cancer samples were overlapped with 0% methylated calibrator.The methylation levels of 5 cancer samples ranged between 0% and 30%.The methylation levels of 29 cancer samples ranged between 31% and 80%.The methylation levels of 20 cancer samples ranged between 81% and 100%.HRM could be used to reliably detect the as low as 10% methylation for each assay,whereas methylation specific PCR(MSP) could be used to detect 30% methylation level.No significant association between methylation level and patients' age(X~2=3.29,P=0.19),PSA level(X~2=2.04,P=0.36) was found.However,DLC-1 methylation was higher in the prostate cancer tissues with advanced TNM stage(X~2=9.04,P=.01).Conclusions The quantitative method for DLC-1 methylation with HRM is successfully established.It is convenient with good reproducibility and high sensitivity.DLC-1 methylation could be used as the molecular marker for estimation of malignancy in prostate cancer.

Objective To explore the expressions and clinical significance of epidermal growth factor receptor (EGFR),COOH-terminus tensin-like molecule (CTEN)and epithelial cadherin (E-cad)in non-small cell lung cancer (NSCLC).Methods The expressions of EGFR,CTEN and E-cad in 36 cases of normal lung tissue and 82 cases of NSCLC tissues were observed with immunohistochemical SP method and their correlation with NSCLC invasion,metastasis and prognosis was analyzed.Results The positive rate of EGFR,CTEN and E-cad was 58.5% (48/82),69.5% (57/82)and 28.1% (23/82)in 82 cases of lung cancer,while 13.9% (5/36),0.0% (0/36)and 100% (36/36)in normal tissues;the differences were all significant (P<0.05).EGFR,CTEN and E-cad expressions in NSCLC tissues were significantly correlated with tumor differentiation,TNM stage and lymph node metastasis (P<0 .0 5 ).The expressions of EGFR and CTEN were correlated with each other in NSCLC (r=0 .5 3 0 , P<0.001),while the expressions of EGFR and CTEN were correlated with that of E-cad (r=0.499,P<0.001;r=0.333,P=0.001 ).Multivariate Cox regression analysis showed that CTEN expression in NSCLC was an independent prognostic factor (P<0.05 ).Conclusion EGFR,CTEN and E-cad may play a role in the development,invasion and metastasis of NSCLC and have some significance in predicting the prognosis of NSCLC.

Objective: To evaluate the inter-bottle variations, stability in consumption and interchangeability of unassayed biochemistry serum controls. Methods: Comparison between unassayed serum controls from a domestic "Pretrol^®" and an international "Bio-Rad" manufacturer was conducted in department of laboratory, in May 2016, with Roche Cobas 8000, Roche Hitachi 7600 and Siemens 2400 modular analyzer. The inter-bottle variation was determined by monitoring the inter-batch variation of 10 bottles of control samples after eliminating the intra-batch variation from the same bottle. Stability in consumption was determined as the precision after 7 days storage under 2 ℃ to 8 ℃ or 30 days of storage under -20 ℃ since reconstitution. The interchangeability was determined as the precisionbetween the controls from different manufacturers for the same test. Results: The inter-bottle imprecision of controls from domestic manufacturer for 13 biochemistry tests (CV concentration 1/CV concentration 2) were potassium (0.26%/0.42%), sodium (0.26%/0.21%), phosphorus (0.00%/0.62%), cholesterol (0.56%/0.54%), total protein (0.52%/0.33%), albumin (0.44%/2.00%), alanine aminotransferase (1.72%/0.57%), γ-glutamylaminotransferase (0.52%/0.62%), aspartate aminotransferase (3.10%/1.09%), lactate dehydrogenase (0.76%/0.91%), alkaline phosphatase (1.13%/0.97%), amylase (0.30%/0.39%) and glucose (0.00%/0.40%). The stability in consumption of the controls from the domestic manufacturer (CV concentration 1/CV concentration 2 under 2 ℃ to 8 ℃storage; CV concentration 1/CV concentration 2 under -20 ℃ storage) were potassium (1.06%/0.36%; 0.74%/0.48%), sodium (0.49%/0.59%; 0.72%/0.65%), phosphorus (0.95%/0.80%; 1.43%/0.84%), cholesterol (1.49%/1.58%; 2.17%/1.80%), total protein (0.84%/0.75%; 1.60%/1.68%), albumin (1.33%/2.28%; 1.94%/2.43%), alanine aminotransferase (1.41%/0.51%; 3.24%/1.60%) γ-glutamylaminotransferase (1.16%/1.16%; 2.85%/2.49%), aspartate aminotransferase (4.37%/2.14%; 2.99%/1.31%), lactate dehydrogenase (2.70%/2.54%; 3.84%/2.97%), alkaline phosphatase (2.63%/1.96%; 2.31%/2.10%), amylase (0.95%/2.19%; 1.58%/1.38%) and glucose (0.60%/0.48%; 1.41%/1.55%). The Inter-bottle variation and stability in consumption of biochemistry test unassayed controls from the domestic manufacturer were compatible for clinical assay according to the CV% specification from the Clinical Biochemistry Test Quality Requirement (WS/T 403-2012). The imprecision of the controls from both the domestic and international manufacturers (CVp concentration 1/CVp concentration2; CVq concentration 1/CVq concentration 2) were potassium (0.52%/0.46%; 2.39%/0.47%), sodium (0.30%/0.17%; 0.81%/0.47%), phosphorus (2.72%/1.11%; 4.57%/2.07%), cholesterol (0.29%/1.38%; 2.94%/1.81%), total protein (0.66%/2.46%; 1.85%/2.54%), alkaline phosphatase (2.67%/4.66%; 3.58%/8.55%), total bilirubin (5.71%/5.09%; 9.55%/7.41%), albumin (1.10%/2.61%; 4.79%/1.93%), alanine aminotransferase (6.42%/1.25%; 5.74%/1.63%), γ-glutamylaminotransferase (2.27%/4.35%; 4.38%/0.74%), aspartate aminotransferase (0.56%/2.84%; 0.91%/2.11%) and lactate dehydrogenase (2.36%/2.47%; 3.10%/1.52%). The interchangeability of serum controls from domestic manufacturer was better than clinical serum samples. Conclusion The unassayed serum biochemistry test controls from domestic manufacturer are suitable for the intra-laboratory quality control and showed a promising compatibility for inter-laboratory quality control usage.