[Differential plasma protein profiles in patients with hyperlipidemia & atherosclerosis of different patterns of phlegm-stasis syndrome].

OBJECTIVE: To investigate the differential plasma protein profiles in patients with hyperlipidemia & atherosclerosis (H&A) of different patterns of phlegm-stasis syndrome (PSS) for seeking their biomarker proteins. METHODS: Two-dimensional gel electrophoresis and gel screening graphical analysis were performed on plasma proteins got from 146 patients; corresponding protein spots were fetched from the gel for two-stage mass-spectrometric analysis by quadruple time-of-flight mass spectrometry; then the differential proteins for PSS were discriminated by Fisher discriminate analysis. RESULTS: Excepting two uncertain proteins, 7 differential proteins were screened out from the 11 differentially expressed plasma protein spots with variability over 100% in the inter-block matching. Classic analysis found that haptoglobin precursor and fibrinogen gamma chain were possibly the plasma biomarker proteins for H & A; fibrinogen beta chain and apolipoprotein A-I precursor were that set apart PSS from non-PSS; fibrinogen gamma chain, albumin and apolipoprotein A-I precursor were for phlegm syndrome; haptoglobin precursor, adrenomedullin binding protein precursor, albumin and complement component C4 were for stasis syndrome; albumin and adrenomedullin binding protein precursor were for the phlegm-stasis mutual blocking syndrome. Moreover, the above mentioned expressions of possible marker proteins had their own special rule of changing in the transforming progress of PSS. CONCLUSION: This study reported, for the first time, the existence of evident variation of functional protein constitution in different patterns of PSS, and definite compatibility being detected in some functional proteins, which may be the marker proteins for making diagnosis and prognosis of PSS in H&A. Besides, preliminary proof for the transformation of PSS has gained at the functional protein level.

[Relationship between plasma protein expression profiles and states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis].

OBJECTIVE: To investigate the relationship between the plasma biomarker proteins and the states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis. METHODS: The states of Zang-Fu organs in 146 patients with hyperlipidemia and atherosclerosis were diagnosed by syndrome differentiation of traditional Chinese medicine. The plasma proteins from these patients were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Differential protein profiling was established by Image Master 6.0 software, and the differential proteins were analyzed by quadrupole time of flight mass spectrometry (Q-TOF-MS). The association between the plasma biomarker proteins and the states of Zang-Fu organs was analyzed by graphical models. RESULTS: The biomarker proteins such as fibrinogen gamma chain, albumin and apolipoprotein AI (precursor) in discrimination of the patients with phlegm syndrome from phlegm accumulating with stagnation syndrome were correlated with the deficiency of kidney-qi, heart-qi and spleen-qi. Among the four biomarker proteins in discrimination of the patients with phlegm syndrome from blood stagnation syndrome, albumin, adrenomedullin binding protein (precursor) and haptoglobin (precursor) were correlated with the deficiency of kidney-qi and heart-qi, but complement component C4 was independent of the deficient Zang-Fu organs. The biomarker albumin was associated with the deficiency of kidney-qi, heart-qi and spleen-qi, and adrenomedullin binding protein (precursor) was correlated with the deficiency of spleen-qi in discrimination of the patients with blood stagnation syndrome from phlegm accumulating with stagnation syndrome. As the potential biomarker proteins in discrimination of the patients with non-phlegm and non-stagnation syndrome from phlegm accumulating with stagnation syndrome, the fibrinogen beta chain was related with the deficiency of kidney-qi, and apolipoprotein AI (precursor) was correlated with both the deficiency of kidney-qi and heart-qi. CONCLUSION: There exists inherent correlation between the states of Zang-Fu organs and the plasma probable biomarker proteins in the patients with different phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis.

[Applying matching matrix, factor analysis and clustering analysis to investigation of characteristics of syndromes of phlegm and blood stasis in patients with coronary heart disease].

OBJECTIVE: To investigate the characteristics of syndromes of phlegm and blood stasis in patients with coronary heart disease by multiple statistical methods of matching matrix, factor analysis and clustering analysis, and to provide some references for classification and normalization of diagnosis of syndromes of phlegm and blood stasis of coronary heart disease. METHODS: The correlations among 46 kinds of symptoms in syndrome of non-phlegm and non-blood stasis, syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking in 200 patients with coronary heart disease were analyzed by matching matrix, factor analysis and clustering analysis. RESULTS: The manifestations of tongue and pulse in syndromes of phlegm and blood stasis were significantly different from those in syndrome of non-phlegm and non-blood stasis. The pathogenesis of viscera in syndromes of phlegm and blood stasis lied in the heart and kidney, and the syndrome of deficiency of heart qi was the most common one while the syndrome of deficiency of kidney qi took the secondary place. The syndrome of phlegm was often accompanied by syndrome of deficiency of spleen qi. Only 15 ones of 46 clinical symptoms showed high frequency in concomitant appearance in syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking. Apart from having the common symptoms in syndrome of deficiency in origin, the syndrome of phlegm especially showed white and greasy fur and slippery pulse as well as distention and fullness of chest and abdominal distension; the syndrome of blood stasis showed purplish tongue and ecchymosis on tongue as well as fixed pain; and the syndrome of phlegm-blood stasis blocking showed the main symptoms of both syndrome of phlegm and syndrome of blood stasis. CONCLUSION: The statistical methods of matching matrix, factor analysis and clustering analysis are convenient, and can definitely indicate the clinical characteristics and syndrome differentiation of viscera of different syndromes of phlegm and blood stasis, which are beneficial to further research of diagnosis and differentiation of such syndromes of coronary heart disease.

[Study on relationship between polymorphism of apolipoprotein E gene and syndromes of phlegm and blood stasis in patients with coronary heart disease].

OBJECTIVE: To explore the relationship between polymorphism of apolipoprotein E (ApoE) exon 4 gene and different syndromes in patients with coronary heart disease (CHD). METHODS: Two hundred patients with CHD were divided into four groups according to syndrome differentiation, including syndrome of phlegm (PS), syndrome of blood stasis (BSS), syndrome of phlegm-blood stasis blocking (PBBS) and syndrome of non-phlegm and non-blood stasis (NPNBS). One hundred healthy volunteers were included in control group. Blood lipids were measured by routine examination. Total DNA of peripheral blood was extracted. ApoE genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. All data were analyzed by SAS software. RESULTS: (1)The occurrence rate of epsilon4 allele of ApoE in patients with CHD was 19.5%, significantly higher than 9.5% in the control group (P<0.05), and the E 3/4 genotype was especially more frequent (P<0.01). (2) The levels of total cholesterol (TC), total triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in patients with epsilon4 were higher than those in patients without epsilon4 (P<0.01). (3) The frequencies of epsilon4 allele and E3/4 genotype in patients with PS were significantly higher than those in patients with BSS (P<0.05). CONCLUSION: ApoE epsilon4 allele, especially E3/4 genotype, is the risk factor of CHD. There is a relatively close relationship between patients with ApoE epsilon4 allele and PS. It may be one of the main susceptible genes in CHD patients with PS.