Study on the Tongue Manifestations for the Blood-Stasis and Toxin Syndrome in the Stable Patients of Coronary Heart Disease

Objective： To explore the tongue manifestations for the blood-stasis and toxin syndrome in the stable patients of coronary heart disease （CHD） according to the acute cardiovascular events （ACEs） in one-year follow-up which based on the pathogenesis hypothesis of ＂blood-stasis and toxin causing catastrophe＂. Methods： Totally 254 stable CHD cases were enrolled after diagnosed by coronary angiography, their tongue appearances were recorded by the digital camera of uniform type, 29 cases with ACEs during one-year follow-up were assigned in ACEs group. The non-ACEs were matched in proportion of 2：1 according to the gender, age （± 2.5 years）, diabetes mellitus history and previous acute coronary syndrome hospitalization history in the non-ACEs group, and 54 cases were eligibly included. The differences of tongue appearance between the ACEs and non-ACEs group were compared. Results： Fifteen cases manifested with bluish tongue （including bluish-grounding or bluish purple tongue）, among which 11 cases （37.9%） in the ACEs group and four cases （7.4%） in the non-ACEs group, and there was significant difference （P〈0.002）. Twenty six cases showed yellow tongue coating in the non-ACEs group, which was significantly higher than that in the ACEs group （48.1 vs. 10.3%, P=0.001）. The tongue of sticky greasy coating was more frequently occurred in the non-ACEs group than that in the ACEs group （66.7% vs. 41.4%, P=0.026）. The proportion of purplish-red sublingual vessel was higher in the ACEs group than that in the non-ACEs group （41.4% vs. 20.4%, P=0.041）. Odd ratio （OR） analysis showed that the patients with bluish tongue, purplish-red sublingual vessel, dry-greasy or dirty greasy coating were more likely to experience ACEs during one- year follow-up （OR： 11.67, 95%CI： 3.34-40.81, P〈0.001; OR： 2.76, 95%CI： 1.02-7.44, P〈0.05; OR： 3.12, 95%, CI： 0.89-10.92, P=0.066）. Conclusions： The bluish tongue （including bluish-grounding or bluish purple tongue） and purplish-red sublingual vessel were potential tongue manifestations of blood-stasis and toxin. The tongue coating changing from sticky greasy to dry greasy or dirty greasy was also probably a tongue manifestation of ＂transforming toxin＂, which need demonstration by further study.

Relationship between Platelet Activation Related Factors and Polymorphism of Related Genes in Patients with Coronary Heart Disease of Blood-stasis Syndrome

Objective： To comparatively study the expressive conditions of platelet activation related factors （GPⅠb, GPⅡb-Ⅲa and GMP-140） in healthy subjects and patients with coronary heart disease （CHD） of blood-stasis （BS） or non-blood-stasis （non-BS） syndrome, and to analyze the relationship between the activities of various glycoproteins and the polymorphism of genes. Methods： With case control design adopted, patients with the CHD （40 of BS, 37 of non-BS） and 39 healthy subjects for control, all fitting to the inclusion criteria, were selected in this study. The number of affected coronary branches was recorded by the contrast examination. The mean fluorescence intensity （MFI） of GPⅠb, GPⅡb-Ⅲa, and GMP-140 （CD42b, CD61, CD62p） in patients and healthy persons was measured with flow cytometry, the polymorphism of HPA-3 gene was detected by Taqman probe technique and that of HPA-2 gene was determined by gene sequencing. Results： MFI of CD61 and CD62p was higher in the CHD patients than in the healthy control, which was also higher in patients of BS syndrome than in patients of non-BS syndrome （P〈0.05）; MFI of CD42b was lower in the CHD patients than in the healthy control （P〈0.05）, but showing insignificant difference between BS and non-BS syndrome （P〉0.05）; at the same time, no significant difference of all the above-mentioned three MFI could be found in patients with various numbers of affected coronary branches, neither in patients with different genotypes at GPⅡb HPA-3 and GPⅠb HPA-2 polymorphism loci （P〉0.05）. Conclusion： （1） The activities of GP Ⅱ b-Ⅲa and GMP-140 were obviously increased in the genesis and developing process of CHD and CHD of BS syndrome, and so they could be taken as one of the objective indexes for microscopic diagnosis of BS syndrome. （2） The level of GPⅠb was lower in CHD patients than in healthy persons, but it was not a sensitive indicator for BS syndrome of CHD. （3） Levels of GP Ⅱb-Ⅲa, GPⅠb and GMP-140 were not related with the number of affected coronary branches in CHD patients. （4） The changes in amino-acids expression induced by the two loci brought no significant influence on GPⅠb and GP Ⅱb-Ⅲa activities.

The Diagnostic Criteria of Blood-Stasis Syndrome:Considerations for Standardization of Pattern Identification

Pattern identification （PI）, also called Bian Zheng （辨证）, syndrome differentiation, pattem diagnosis, or pattem classification, is the basic principle and the key concept of Chinese medicine （CM）. The core of PI is CM syndrome, on which CM theory, therapeutic method, prescribing formula and the use of Chinese herbal medicine are basically based. PI, in fact, is another classification method anticipated to improve the clinical efficacy. How to make an exact PI seems to be very important for taking full advantage of PI in clinical practice. Therefore, the establishment of diagnostic criterion of pattern has been the prerequisite for the standardization of PI. In recent years, a lot of diagnostic criteria of different CM patterns have been formulated. Taking the diagnostic criteria for blood-stasis syndrome as a model, the methodologies and considerations in establishing a pattern diagnostic criterion were discussed in this paper, which might be of great reference value in future PI standardization research.

Expression of T-Helper 17 Cells and Signal Transducers in Patients with Psoriasis Vulgaris of Blood-Heat Syndrome and Blood-Stasis Syndrome

Objective： To investigate the levels of cytokines related to T-helper （Th） 17 cells in serum and signal transducers in the psoriatic lesions of patients with psoriasis vulgaris of blood-heat syndrome （BHS） and blood-stasis syndrome （BSS）. Methods： Sixty patients with psoriasis vulgaris were divided into the BHS and BSS groups according to the syndrome differentiation of Chinese medicine （CM）. Ten healthy subjects were considered as the control group. Cytokine levels of interleukin （IL）-17, IL-23 and IL-6 in serum were determined by enzyme-linked immunosorbent assay. Expression levels of signal transducer and activator of transcription 3 （STAT3）, p38-mitogen-activated protein kinase （MAPK） and STAT6 in the psoriatic lesions were determined using immunohistochemistry （IHC）, Western blot, and real-time quantitative reverse transcription polymerase chain reaction, respectively. Results： Production of IL-17, IL-23 and IL-6 in the BHS group and BSS group were significantly increased compared with those in the control group （P〈0.05）. Levels of IL-17 and IL-23 in the BHS group were higher than those in the BSS group （P〈0.05）. Compared with the control group, IHC positive expressions and protein expressions of STAT3 and p38-MAPK, and the STAT3 mRNA expressions in the BHS and BSS groups were significantly higher （P〈0.05 or P〈0.01）. The protein expression of STAT3 in the BHS group was significantly higher than that in the BSS group （P〈0.05）. Conclusions： Cytokines in serum and signal transducers in the psoriatic lesions alter with various CM syndromes of psoriasis. The results provide scientific basis for the treatment based on syndrome differentiation of CM in treating psoriasis vulgaris.

Clinical Observation on the Effect of Zengse Pill (增色丸) in Treating Patients with Vitiligo of Qi-stagnancy and Blood-stasis Syndrome Type

Objective： To observe the clinical efficacy of Zengse Pill （增色丸, ZSP） on patients with vitiligo of qi-stagnancy and blood-stasis syndrome type （V-QB）, and to preliminarily explore its mechanism of action. Methods： Sixty-five V-QB patients, with their diagnosis confirmed by clinical examination, were randomized by digital table method into two groups, with 31 patients in the control group and 34 in the treatment group. Cobamamide （2 tablets） was administered orally to all patients, and Psoralea tincture （a self-formulated preparation） was applied externally thrice a day. In addition, for patients in the treatment group, ZSP was given orally, at 5 pills per dose, 3 times every day. The therapeutic course for both groups was 3 months. Patients were re-examined every half-month, and changes in the skin lesions were observed and recorded. The levels of lymphocyte subsets, serum immune globulin, and complement C3 and C4 in patients were determined before and after the therapeutic course and compared with the corresponding indexes determined in 21 healthy subjects, Results： The total effective rate in the treatment group was 82.4%, which was markedly higher than that in the control group （54.8%）, showing a significant difference （P〈O.05）. After treatment, CD4^＋ percentage, CD4^＋/CD8^＋ratio, and blood levels of C3 and C4 increased, while CD8^＋ percentage decreased in the treatment group （P〈0.05 or P〈0.01）. All these indexes remained unchanged in the control group, and the respective comparisons between groups showed significant differences （P〈0.01）. Conclusion： ZSP has a definite clinical effect on the treatment of V-QB but with no evident adverse reactions, and it can increase the CD4^＋ percentage, CD4^＋/CD8^＋ ratio, and the levels of serum C3 and C4, thus regulating the immunity of the organism, which might be one of its mechanisms of action.

Transforming growth factor β1 is a differentially expressed candidate protein of congestive heart failure with Qi-deficiency-blood-stasis syndrome

OBJECTIVE: To investigate the role of tongue coating fluid protein in regulation of congestive heart failure(CHF) in Qi-deficiency-blood-stasis syndrome.METHODS: We studied patients with CHF(3 patients with Qi-deficiency-blood-stasis syndrome and 3 without Qi-deficiency-blood-stasis syndrome) to investigate differentially expressed proteins. We also included a control group. A biotin label-based antibody array was used for testing tongue coating fluid samples from patients. Net-work analysis of these differentially expressed proteins was conducted using the STRING database,which can predict the relations between differentially expressed proteins and CHF with Qi-deficiency-blood-stasis syndrome.RESULTS: A total of seven differentially expressed proteins were identified, and among these, transforming growth factor β1(TGF-β1) gets a particular attention for us has drawn specific attention.Network analysis showed a homologous relationship of TGF-β1 with bone morphogenetic protein15, which is associated with myocardial fibrosis.CONCLUSION: Occurrence and development of CHF may result from certain DE-proteins and associated signaling pathways. TGF-β1 protein may be a candidate marker for assessing the risk of CHF in Qideficiency-blood-stasis syndrome.

Differential Expression of MicroRNA in Endothelial Cells Incubated with Serum of Hypertension Patients with Blood-Stasis Syndrome

Objective： To screen out blood-stasis syndrome （BSS）-associated microRNA and therefore determine the possible target for treating hypertension. Methods： A high-energy sequencing method and digital gene expression sequencing theory were adopted to sequence microRNA （miRNA） and messenger RNA （mRNA）, and to determine differential expression in human umbilical vein endothelial cells incubated with serum samples from hypertension patients with or without BSS, and healthy controls. The results were confirmed using gene prediction software. Results： A total of 13 miRNAs and 11 mRNAs showed statistical difference both in the BSS/normal groups and BSS/non-BSS groups, respectively. Four pairs of target mRNNmiRNA were identified： FRMD4Nhsa-miR-34a, MAP3K14/hsa-miR-34a, PER1/hsa-miR-34a, and FGF2/hsa-miR-132. Conclusion： Four mRNNmiRNA pairs mentioned above seem to be involved in pathogenesis and maintenance of hypertension with BSS.

Acupuncture combined with medication for blood-stasis in unstable angina patients with different thrombolysis in myocardial infarction risk stratification

Objective To observe the intervention effects of acupuncture combined with standardized treatment of western medicine on blood-stasis syndrome in unstable angina(UA)patients with different thrombolysis in myocardial infarction(TIMI)risk stratification.Methods According to TIMI risk score,a total of 72 UA patients were included,24 cases in low-risk(0 to 2 points)group,24

Effects of acetylcysteine on micro-inflammation and pulmonary ventilation in chronic obstructive pulmonary disease exacerbation

BACKGROUND Acute exacerbation of chronic obstructive pulmonary disease(AECOPD)is a serious complication of chronic obstructive pulmonary disease,often characterized by increased morbidity and mortality.In traditional Chinese medicine,AECOPD is linked to phlegm-heat and blood-stasis,presenting symptoms like thick sputum,fever,and chest pain.It has been shown that acetylcysteine inhalation in conjunction with conventional therapy significantly reduced inflammatory markers and improved lung function parameters in patients with AECOPD,suggesting that acetylcysteine may be an important adjunctive therapy for patients with phlegm-heat-blood stasis type AECOPD.AIM To investigate the effect of acetylcysteine on microinflammation and lung ventilation in patients with phlegm-heat and blood-stasis-type AECOPD.METHODS One hundred patients with phlegm-heat and blood-stasis-type AECOPD were randomly assigned to two groups.The treatment group received acetylcysteine inhalation(10%solution,5 mL,twice daily)along with conventional therapy,whereas the control group received only conventional therapy.The treatment duration was 14 d.Inflammatory markers(C-reactive protein,interleukin-6,and tumor necrosis factor-alpha)in the serum and sputum as well as lung function parameters(forced expiratory volume in one second,forced vital capacity,and peak expiratory flow)were assessed pre-and post-treatment.Acetylcysteine inhalation led to significant reductions in inflammatory markers and improvements in lung function parameters compared to those in the control group(P<0.05).This suggests that acetylcysteine could serve as an effective adjunct therapy for patients with phlegm-heat and blood-stasis-type AECOPD.RESULTS Acetylcysteine inhalation significantly reduced inflammatory markers in the serum and sputum and improved lung ventilation function parameters in patients with phlegm-heat and blood-stasis type AECOPD compared with the control group.These differences were statistically significant(P<0.05).The study concluded that acetylcysteine inhalation had a positive effect on microinflammation and lung ventilation function in patients with this type of AECOPD,suggesting its potential as an adjuvant therapy for such cases.CONCLUSION Acetylcysteine inhalation demonstrated significant improvements in reducing inflammatory markers in the serum and sputum,as well as enhancing lung ventilation function parameters in patients with phlegm-heat and bloodstasis type AECOPD.These findings suggest that acetylcysteine could serve as a valuable adjuvant therapy for individuals with this specific type of AECOPD,offering benefits for managing microinflammation and optimizing lung function.

Effects of L.F04,the Active Fraction of Lycopus lucidus, on Erythrocytes Rheological Property

Objective: To study the effects of L.F04, the active fraction of Lycopus lucidus, on erythrocytes rheological property so as to investigate its mechanism in promoting blood circulation and removing blood stasis. Method: The effects of L.F04 (used for treatment for 10 days in different dosages) on deformability, aggregation and membrane liquidity of erythrocytes (MLE) as well as whole blood apparent viscosity (η_b) were examined on the basis of rat model of blood-stasis syndrome induced by venous injection of high molecular weight dextran. Result: As compared with the normal control group, the model group's RBC deformability and MLE were lower, and the aggregation of erythrocytes and η_b were higher. Compared with the model group, both L.F04 0.612g/kg and 0.306g/kg showed significant effect in improving deformability and inhibiting aggregation of red blood cells (RBC) and reducing blood viscosity. The trend of improving MLE was also shown. Conclusion: L.F04 could significantly improve the abnormal rheological property of erythrocytes.

Differential Gene Expression Profiles in Coronary Heart Disease Patients of Blood Stasis Syndrome in Traditional Chinese Medicine and Clinical Role of Target Gene

Objective:To investigate the differential gene expression profiles in coronary heart disease(CHD) patients of blood-stasis syndrome(BSS) by oligonucleotide microarray technique,and the clinical significance of target gene.Methods:Subjects were assigned to CHD patients with BSS(n=8),CHD patients without BSS (n=8),and BSS patients without CHD(n=8) based on coronary angiography and the diagnostic criteria of BSS. The sex- and age-matched healthy volunteers(n=8) were enrolled as the control group.Venous blood s...

Circulating Levels of Inflammatory Cytokines in Patients with Psoriasis Vulgaris of Different Chinese Medicine Syndromes

Objective： To investigate whether the serum levels of inflammation-related cytokines might be different between the healthy individuals and the psoriatic patients diagnosed of three varied Chinese medicine （CM） syndromes [blood-stasis syndrome （BSS）, blood-dryness syndrome （BDS） and wind-heat syndrome （WHS）]. Methods： A total of 62 psoriatic patients were recruited and assigned to 3 groups according to their CM syndromes, including 27 patients of BSS, 21 of BDS and 14 of WHS. Another 20 sex- and age- matched healthy subjects were enrolled into the control group. Serum concentrations of multiple cytokines, including monocyte chemotactic protein-1 （MCP-1）, macrophage inflammatory protein-1α （MIP-1α）, soluble CD40 ligand （SCD40L）, tumor necrosis factor-α （TNF-α）, epidermal growth factor （EGF）, interleukin-8 （IL-8）, interleukin-17 （IL-17）, interferon γ inducible protein-10 （IP-10） and vascular endothelial growth factor （VEGF）, were measured by a multiplexed flow cytometric assay. Results： The circulating levels of MIP-1α, TNF-α, IL-8, and IP-10 were significantly increased in the psoriatic patients compared with the healthy controls （P〈0.01）. Male and female patients tended to have higher serum levels of MCP-1 and IP-10, respectively （P〈0.05）. Interestingly, compared with the control group, 6 out of the 9 cytokines （MCP-1, MIP-1α, TNF-α, EGF, IL-8 and IP-10） were substantially increased in the BSS group （P〈0.05 or P〈0.01）, whereas only MIP-1α and IL-8 levels were elevated in the BDS group （P〈0.05 or P〈0.01） concurrent with lowered concentrations of SCD40L and IL-17 （P〈0.05）. In the WHS group, MIP-1α was the only cytokine whose level was evidently increased （P〈0.01）, in contrast to IL-17 which was decreased as compared with the control （P〈0.05）. The psoriatic patients overall owned higher levels of MIP-1 a and IL-8 in the circulation which were comparable among the 3 groups of CM syndromes （P〈0.01）. In contrast, TNF-α level of the BSS group was the highest among the three （P〈0.01）, followed by the BDS and the WHS groups. Conclusions： The expression profiles of cytokines in the circulation might not be necessarily identical for psoriatic patients with different CM syndromes. Accordingly, the serum concentrations of certain cytokines could potentially be used as the ancillary indices for the clinical classification of psoriatic CM syndromes.

Study on the Relationship between Blood Stasis Syndrome and Clinical Pathology in 227 Patients with Primary Glomerular Disease

To investigate the relationship between the severity of Chinese medicine(CM) bloodstasis syndrome(BSS) with clinical features and renal lesion indexes of the primary glomerular disease. Methods:An epidemiological survey was conducted to collect the data of 227 patients diagnosed as chronic primary glomerular diseases,and their severity of BSS were scored three days before renal biopsies were performed.The following clinical indexes were analyzed:age,course of glomerular diseases,24-h urine protein ration(Up...

Traditional Chinese Medicines alleviated myocardial ischemia by regulating Qi and promoting blood circulation

OBJECTIVE:To investigate the efficacy of Chinese medicines on Qi stagnation and blood stasis in rats with myocardial ischemia.METHODS:Fifty male Wistar rats were randomly divided into five groups(n=10)as follows:(a)sham operation(Sham),(b)myocardial ischemia(Model),(c)treatment that regulates Qi(Qi),(d)treatment that promotes blood circulation(Blood),(e)treatment that both regulates Qi and promotes blood circulation(QB).The rat model was established via activities restriction for 6 h followed by tail clamp stimulation for 5 mins every day for 7 d and occlusion left coronary anterior descending artery.Afterwards rats were treated with medicines that regulate Qi and/or promote blood circulation via gavage for 14 d.Behavioral parameters were evaluated using open field and elevated plus-maze tests.The tongue color and sublingual vein were visually examined.Blood flow perfusion of tongue and auricle were detected using PIMⅡ.The mesenteric microcirculation was examined via capillaroscopy,and hemodynamics was assessed using a polygraph system.Serum homocysteine(Hcy),creatine kinase isoenzyme(CKMB)levels and endothelin-1(ET-1)were measured.Hematoxylin and eosin staining and transmission electron microscopy were employed to detect the myocardial morphology and ultrastructure,respectively.RESULTS:Compared with findings in Sham group,rats in model group had coarse hair,dark mucosa of the lips and claw,low activity,and increased anxiety.Compared with findings in Model group,rats in the three treatment groups exhibited a lighter tongue color without an extended and varicose sublingual vein.There were significant increases of auricle blood flow perfusion in the Qi group and tongue bottom blood flow perfusion in the QB group.Compared with findings in Model rats,rats in Blood group exhibited improved mesenteric microcirculation associated with increased mesenteric blood flow and a larger arteriole diameter.Moreover,compared with findings in Model rats,Qi and QB rats exhibited increased left ventricular±dp/dtmax,decreased serum CKMB,Hcy,ET-1 levels,and reduced myocardial ultrastructural damage.CONCLUSION:Myocardial ischemia damage was suppressed by Traditional Chinese Medicines that regulate Qi and promote blood circulation.

Constructing protein-protein interaction network of hypertension with blood stasis syndrome via digital gene expression sequencing and database mining

OBJECTIVE： To construct a protein-protein interaction（PPI） network in hypertension patients with blood-stasis syndrome（BSS） by using digital gene expression（DGE） sequencing and database mining techniques.METHODS： DGE analysis based on the Solexa Genome Analyzer platform was performed on vascular endothelial cells incubated with serum of hypertension patients with BSS. The differentially expressed genes were f iltered by comparing the expression levels between the different experimental groups. Then functional categories and e nriched pathways of the unique genes for BSS were analyzed using Database for Annotation, Visualization and Integrated Discovery（DAVID） to select those in the enrichment pathways. I nterologous Interaction Database（I2D） was used to construct PPI networks with the selected genes for hypertension patients with BSS. The potential candidate genes related to BSS were identif ied by comparing the number of relationships among genes. Confi rmed by quantitative reverse transcription-polymerase chain reaction（q RTPCR）, gene ontology（GO） analysis was used to infer the functional annotations of the potential candidate genes for BSS.RESULTS： With gene enrichment analysis using DAVID, a list of 58 genes was chosen from the unique genes. The selected 58 genes were analyzed using I2 D, and a PPI network was constructed. Based on the network analysis results, candidate genes for BSS were identifi ed：DDIT3, JUN, HSPA8, NFIL3, HSPA5, HIST2H2 BE, H3F3 B, CEBPB, SAT1 and GADD45 A. Verif ied through qRT-PCR and analyzed by GO, the functional annotations of the potential candidate genes were explored.CONCLUSION： Compared with previous methodologies reported in the literature, the present DGE analysis and data mining method have shown a great improvement in analyzing BSS.

Development of a valid and reliable seven emotions impairment questionnaire and assessment of its predictability for phlegm and blood stasis

OBJECTIVE:To develop and validate a Seven Emotions Impairment questionnaire(SEIQ),to define an optimum cut-off point for the SEIQ,and to examine whether SEI was predictive of Phlegm and Blood Stasis(BS).METHODS:Two hundred outpatients and 75 college students were asked to complete the SEIQ,the Profile of Mood States(POMS),Phlegm Pattern Questionnaire(PPQ),and BS Questionnaire(BSQ).Twelve clinicians determined whether the outpatients exhibited SEI.SEIQ data were used to examine the internal consistency and determine validity for the outpatients.SEIQ,POMS,PPQ,and BSQ data were used to examine concurrent validity and predictability of SEI for Phlegm and BS in the college students.Total SEIQ scores and the clinicians' diagnoses of the outpatients were considered to define an optimum cut-off score for the SEIQ.RESULTS:The 18-item SEIQ had satisfactory internal consistency(α = 0.905) and concurrent validity.In the construct validity test,four factors(chest-anxiety,fatigue-depression,working-family-troubles,and sleep-memory) were identified.In the receiver operator characteristic curve curve analysis,the sensitivity,specificity,and area under the curve of the SEIQ were 67.2%,72.1%,and 73%,respectively.The optimum cut-off score was defined as nine points.SEIQ scores were strongly predictive of Phlegm and BS(β = 0.862 and 0.673,respectively).CONCLUSION:Based on our results,we concluded that the SEIQ is a reliable and valid instrument for evaluating SEI,and is strongly predictive of Phlegm and BS.