Syndrome Distribution And GC-MS Based Metabonomic Studies On Pleural Effusions

Objective:To investigate the distribution of the syndrome distribution and classification of pleural effusions and compare the malignant pleural effusion with benign one in the TCM syndrome to improve the clinical treatment based on syndrome distribution, and to explore the small molecule biomarkers for the identification of malignant and benign pleural effusions and provide a new insight to diagnose and differentiate malignant and benign ones.Methods:144 pleural effusion patients were enrolled for general data statistical analysis including gender, age and distribution of benign and malignant pleural effusions. According to the principle of TCM syndrome differentiation, all the patients were discriminated from the four types of syndrome. And then, the distribution of the syndrome differentiation and classification were analyzed. Finally, a GC-MS based metabonomic analysis was conducted on 100 pleural effusion samples. Small molecule biomarkers were identified by PCA and OPLS-DA mathematic model, combining with single-variable analysis such as two independent sample T-test, Mann-Whitney U-test.Result:Among the 144 patients with pleural effusion, there are 89 male cases (61.81%) and 55 female cases (38.19%), the onset minimum and maximum age were 19 and 95 years old with 71 cases of malignant pleural effusion (49%),73 patients with benign pleural effusion (51%). The syndrome distribution study showed there were 77 cases of Fluid-retention in the chest hypochondrium syndrome,36 cases of Phlegm and blood stasis syndrome,22 cases of Evil committed chest syndrome,9 cases of Yin asthenia generating instrinsic heat syndrome. The number of four syndrome among the malignant and benign pleural effusion patients was statistically significant (P<0.05). For the further comparison, the number of Fluid-retention in the chest hypochondrium syndrome and Evil committed chest syndrome, Fluid-retention in the chest hypochondrium syndrome and Yin asthenia generating instrinsic heat syndrome, Phlegm and blood stasis syndrome and Evil committed chest syndrome, Phlegm and blood stasis syndrome and Yin asthenia generating instrinsic heat syndrome, among the malignant and benign pleural effusion patients were of statistical significance (P<0.00833), while the number of Fluid-retention in the chest hypochondrium syndrome and Phlegm and blood stasis syndrome, Evil committed chest syndrome and Yin asthenia generating instrinsic heat syndrome were not. Metabonomic study on 100 pleural effusion samples with the multivariate statistical analysis PCA and OPLS-DA model revealed that the benign and malignant pleural effusions were clearly separated by OPLS-DA model. According to the OPLS-DA model, the accuracy rate, sensitivity, specificity of the training and validation group were 96.25%,91.11%,94.71% and 90%,90%, 90% respectively. Combined with single-variable analysis, we found the levels of glycerol, stearic acid, glycerin monostearate, monopalmitin and methylmalonic acid were statistically significant between the malignant and benign pleural effusions (P<0.05). The contents of glycerol, stearic acid, glycerin monostearate, monopalmitin were higher in malignant pleural effusion, while the level of methylmalonic acid was higher in benign one. Furthermore, the pleural effusion patients of the training group were divided into four subgroups according to the syndrome and analyzed with PCA and OPLS-DA models, the OPLS-DA score plot showed the separation trends of the four types of syndrome and the Yin asthenia generating instrinsic heat group were clearly separated from the others.Conclusion:1. The syndrome distribution of the patients with pleural effusion showed that the number of Fluid-retention in the chest hypochondrium syndrome accounted for the most, while the number of Yin asthenia generating instrinsic heat was fewest. We found the syndrome distributions of Fluid-retention in the chest hypochondrium syndrome and Evil committed chest syndrome, Fluid-retention in the chest hypochondrium syndrome and Yin asthenia generating instrinsic heat syndrome, Phlegm and blood stasis syndrome and Evil committed chest syndrome, Phlegm and blood stasis syndrome and Yin asthenia generating instrinsic heat syndrome, were different among the malignant and benign pleural effusion patients. Further, the number of Fluid-retention in the chest hypochondrium syndrome accounted for the most in both malignant and benign pleural effusion patients. However, in malignant pleural effusion patients Phlegm and blood stasis syndrome was more than the benign, while in benign pleural effusion patients Evil committed chest syndrome was more than malignant. The Yin asthenia generating instrinsic heat syndrome only appeared in benign.2. The GC-MS based metabonomic research of pleural effusions displayed high accuracy, sensitivity and specificity in identification of malignant and benign pleural effusion, provided a new measure for clinical diagnosis and differential diagnosis. Glycerol, stearic acid, glycerin monostearate, monopalmitin, methylmalonic acid could be small molecule biomarkers for differential diagnosis of malignant and benign pleural effusions.3. The metabonomic analysis of pleural effusions might be helpful to syndrome differentiation and classification in pleural effusion patients, and the four types of syndrome have the certain trend of separation. The metabolic material basis of all the four types of syndrome needs to be confirmed with expanding the sample size.