1.Level and clinical significance of plasma tissue factor in patient with non-small cell lung cancer
Wanling HUANG ; Yanbin ZHOU ; Yinhuan LI ; Lixia HUANG ; Xingdong CAI ; Qingli ZENG ; Shaoli LI
Clinical Medicine of China 2013;(3):246-249
Objective To detect the plasma level of tissue factor (TF) in non-small cell lung cancer (NSCLC) patients,and to discuss its association with hypercoagulation,venous thromboembolism and prognosis of lung cancer.Methods Sixty-one impatients in our hospital with confirmed lung cancer were enrolled as the study group.Thirteen patients with benign pulmonary diseases and 14 healthy volunteers were selected as the control groups.Bseline and follow-up clinical data were collected from participants.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of TF in plasma of all subjects.Results The levels of TF in plasma from NSCLC patients and participants with benign pulmonary diseases was significantly higher than that in healthy controls((550.88 ± 201.58) ng/L vs (510.77 ± 201.20) ng/L vs (178.34 ±66.73) ng/L,P <0.05).According to the plasma levels of TF,which have been detected in all subjects,the patients were divided into two groups:low level group (range from 103.73 ng/L to 476.22 ng/L) and high level group (range from 476.221 ng/L to 1003.00 ng/L).Statistical analysis showed that there was a positive correlation between plasma TF levels and TNM stages in NSCLC patients (P =0.026).Patient with metastasis had a higher plasma TF level than other patients (P =0.020).The log-rank test revealed that there was no significant difference in survival between the high level group and low level group (x2 =0.145,P =0.704).Multivariate Cox proportional hazards regression analysis indicated that plasma TF levels did not predicted for death(RR =1.001,95%CI0.998-1.004,P=0.452).Conclusion The plasma TF level in NSCLC patients was correlated with TNM stages;it had no significant relationship with hypercoagulation state and survival rate in NSCLC patients.Limitations should be aware of while evaluating the clinical course and prediction of prognosis of NSCLC patients using plasma TF levels.
2.Rapid and high throughput measurement of lipase thermo-stability through ANS fluorescence signal assay.
Weizong FENG ; Junhan LIN ; Shaoli CAI ; Youtu ZOU ; Guoren CHEN ; Ping HUANG ; Yajing LIN ; Bingbing WANG ; Lin LIN
Chinese Journal of Biotechnology 2011;27(4):584-591
We have developed a rapid and high throughput lipase-ANS (8-Anilino-l-naphthalenesulfonic acid) assay to evaluate the thermo-stability of lipases based on the ANS fluorescence signal's increasing and shifting when this small fluorescence probes binds to lipase. The testing lipase samples were incubated at a temperature range of 25 degrees C to 65 degrees C for 30 min before mixed with ANS solution (0.20 mg/mL lipase and 0.05 mmol/L ANS in the buffer of 20 mmol/L Tris-HCl, 100 mmol/L NaCl, pH 7.2) in a cuvette or microplate. Fluorescence signals of the samples were measured at EX 378 nm, EM 465 nm with a fluorescence photometer or a plate reader, and Tm was calculated with the software of GraphPad Prism5.0. The Tm values of several mutants of Penicillium expansum lipase (PEL) were measured with this ANS assay and conventional method simultaneously and the results show that Tm values are comparative and consistent between these methods, suggesting that the lipase-ANS assay is a reliable, rapid and high throughput method for lipase thermo-stability measurement.
Anilino Naphthalenesulfonates
;
chemistry
;
Enzyme Stability
;
High-Throughput Screening Assays
;
methods
;
Hot Temperature
;
Lipase
;
metabolism
;
Spectrometry, Fluorescence