Fecal-associated microbiome differences between phlegm-dampness constitution and balanced constitution

Objective: This study aimed to explore the structural and functional characteristics of the fecalassociated microbiome(FAM) between the phlegm-dampness constitution(PDC) and balanced constitution(BC), and to screen the related specific operational taxonomic unit(OTU) biomarkers.Methods: This was a cross-sectional study. After strictly identifying the constitution of subjects, their clinical index was recorded and counted. Fecal samples were collected for 16S rDNA sequencing. Alpha diversity, beta diversity, and the relative abundance of dominant bacterial taxa were used to describe the FAM structure, and the Wilcoxon rank-sum test, Metagenome Seq, and linear discriminant analysis effect size(LEf Se) were used to screen specific bacterial taxa. Specific OTUs were screened to construct receiver operating characteristic(ROC) curves.Results: Thirty-two subjects were enrolled, including 22 subjects with BC and 10 subjects with PDC.There were significant differences in cold preference, levels of aspartate transaminase, β2-microglobulin,and creatine kinase MB, and alpha diversity indices(Shannon and Shannoneven) between the two groups. In principal coordinate analysis by abund-jaccard distance measure and partial least squares discriminant analysis, bacterial communities clustered separately between the two groups. Furthermore,based on Metagenome Seq, LEf Se, and the Wilcoxon rank-sum test, a total of 43, 18, and 130 OTUs were differentially distributed between BC group and PDC group, respectively, and OTU200, OTU133, and OTU353 were screened when P≤.01. The area under the ROC curve constructed from the 3 selected OTUs was 0.93.Conclusion: The FAM structure and related functional characteristics of the PDC group differed from those of the BC group. In particular, OTU200, OTU133, and OTU353 can be used as unique markers of PDC to assist clinical diagnosis.

Methylation Characteristics of Perivisceral Fat Gene in Obese Rats with Phlegm-dampness Syndrome and the Effect of Wen Dan Decoction

Objective To observe the characteristics of gene methylation in obese rats with phlegm-dampness syndrome induced by the high-fat diet, and to study the effect of Wen Dan Decoction on gene methylation after the intervention. Methods Methylation sites of genes were detected by the MeDIP-seq method. Bioinformatics method was used to analyze the gene methylation characteristics of obesity with phlegmdampness syndrome and the effect of Wen Dan Decoction. Results (1) There were 3 242 methylation differential loci in dietinduced obesity with phlegm-dampness syndrome, of which 1 243 were down-regulated and 1 999 were up-regulated, involving 1 579 differential genes. GO analysis showed that "offactory receptor activity" and others were enriched. The possible signal pathways involved were "Olfactory transduction""Tuberculosis""Systemic lupus erythematosus" and "Ribosome".(2) After the intervention of Wen Dan Decoction in obesity with phlegmdampness syndrome, 4 046 different methylation loci were obtained, including 1 067 down-regulated loci and 2 979 up-regulated loci, involving 2 068 genes. GO analysis showed that "offactory receptor activity" and others were enriched. These genes involved seven signaling pathways, such as "Metabolic pathways".(3) Between diet-induced obesity with phlegm-dampness syndrome and Wen Dan Decoction intervening obesity with the phlegm-dampness syndrome, 582 common genes of methylation differential genes were obtained. After the intervention of Wen Dan Decoction, the number of GO enrichment items was more than that of obesity with phlegm-dampness syndrome, and even the same GO enrichment items involved more genes. Conclusions The phlegm-dampness syndrome of obesityinduced by diet had the characteristics of gene methylation changes, and the intervention of Wen Dan Decoction could also affect the status of gene methylation. The genes affected by Wen Dan Decoction were closely related to the methylation gene of phlegm-dampness syndrome of obesity-induced by diet but covered a wider range.

Proteomic analysis of overweight/obesity and related abnormal glucose and lipid metabolism caused by phlegm-dampness retention

Objective:To investigate the proteomic characteristics of overweight/obesity and related abnormal glucose and lipid metabolism caused by phlegm-dampness retention to identify related biomarkers.Methods:Seventy-one subjects were enrolled in the study.We assessed blood glucose,blood lipids,body mass index(BMI),and phlegm-dampness pattern,which was confirmed by a traditional Chinese medicine clinician.Of the participants,we included healthy participants with normal weight(NW,n=23),overweight/obese participants with normal metabolism(ONM,n=19),overweight/obese participants with pre-diabetes(OPD,n=12),and overweight/obese participants with marginally-elevated blood lipids(OML,n=17).Among them,the ONM,OPD,and OML groups were diagnosed with phlegmdampness pattern.The data-independent acquisition(DIA)method was first used to analyze the plasma protein expression of each group,and the relevant differential proteins of each group were screened.The co-expressed proteins were evaluated by Venn analysis.The pathway analyses of the differential proteins were analyzed using Ingenuity Pathway Analysis(IPA)software.Parallel reaction monitoring(PRM)was used to verify the differential and common proteins in each group.Results:After comparing ONM,OPD,and OML groups with NW group,we identified the differentially expressed proteins(DEPs).Next,we determined the DEPs among OPD,OML,and ONM groups.Using Venn analysis of the DEPs in each group,24 co-expressed proteins were screened.Two co-expressed proteins were verified by PRM.IPA analysis showed that pathways including LXR/RXR activation,acute phase response signaling,and FXR/RXR activation were common to all three groups of phlegmdamp overweight/obesity participants.However,the activation or inhibition of these pathways was different among the three groups.Conclusion:Participants with overweight/obesity have similar proteomic characteristics,though each type shows specific proteomic characteristics.Two co-expressed proteins,VTN and ORM1,are potential biomarkers for glucose and lipid metabolism diseases with overweight/obesity caused by phlegmdampness retention.

Clinical Efficacy of Modified Erchen Decoction on Cervical Spondylotic Vertebral Arteriopathy with Stagnation and Blockade of Phlegm-dampness Syndrome and Effects on Cerebral Blood Flow Parameters

OBJECTIVE: To observe the effect of Modified Erchen Decoction on cervical spondylotic vertebral arteriopathy with stagnation and blockade of phlegm-dampness syndrome and effects on cerebral blood flow parameters. METHODS: A total of 80 cervical spondylotic vertebral arteriopathy(CSA) patients with stagnation and blockade of phlegmdampness syndrome admitted to our hospital from October 2016 to April 2017 were selected and randomly divided into observation group and control group, with 40 cases in each group. The observation group was treated with Modified Erchen Decoction and the control group was given conventional treatment with Western medicine. After 4 weeks of treatment, the main clinical symptoms and signs(vertigo, neck and shoulder pain, headache, psychological and social adjustment, daily life and work) and cerebral blood flow parameters [the peak values of intracranial vertebral-basal artery diastolic blood flow velocity(Vd) and systolic blood flow velocity(Vp)] were compared between the 2 groups. The total clinical effective rate and adverse reactions during treatment were also compared. RESULTS: After treatment, the total effective rate of the observation group was 90.0%, which was significantly higher than that of the control group(77.5%). The difference between the two groups was statistically significant(P < 0.05). After treatment, The scores of vertigo, neck and shoulder pain, headache, psychology and society adaptation, daily life and work were significantly increased(P < 0.05), and the above scores of the observation group were increased more obviously. The difference between the groups was statistically significant(P < 0.05). After treatment, the levels of Vd and Vp were significantly increased(P < 0.05), and the increase of the above indexes was more obvious in the observation group. The difference between the 2 groups was statistically significant(P < 0.05). There were no significant abnormalities in blood routine, urine routine, liver function and renal function. There was no significant difference in the incidence of adverse reactions between the 2 groups.(P > 0.05). CONCLUSION: The treatment of vertebral artery type cervical spondylopathy with stagnation and blockade of phlegm-dampness syndrome by Modified Erchen Decoction can effectively relieve the main clinical symptoms and signs, improve cerebral blood flow parameters, and improve the peak values of vertebral-basal artery diastolic blood flow velocity(Vd) and systolic blood flow velocity(Vp), which is safe and effective, and helps to promote the recovery of cervical function.

Observation on the Curative Effect of"Hand and Foot 12 Needle Acupuncture"on Stroke in Convalescent Period of Phlegm-Dampness Constitution

Objective:To observe the clinical effects of"hand and foot 12 needle acupuncture"on stroke in convalescent period of phlegm-dampness constitution.Methods:The 90 patients were divided into the treatment group(n=45)and the control group(n=45)according to the random number table method.The control group received basic Western medicine treatment+rehabilitation treatment of Western medicine,and the treatment group was additively treated with"hand and foot 12 needle acupuncture"on the basis of the control group.NIHSS score,MRS score and Barthel index(BI)before and after treatment were observed in the two groups,and the clinical efficacy of the two groups was compared.Results:The clinical efficacy of stroke patients in the convalescent period of phlegm-dampness constitution after treatment:the total effective rate of the treatment group was 93.33%,and the total effective rate of the control group was 66.67%.After statistical processing,the clinical efficacy of the treatment group was significantly better than the control group;NIHSS score,MRS score and Barthel index(BI)were better than the control group,and the differences were statistically significant(P<0.05).Conclusion:"Hand and foot 12 needle acupuncture"treatment of phlegm-dampness constitution convalescent stroke clinical effect is remarkable.

Differences in vascular endothelial function and serum proteome between obese people with phlegm-dampness constitution and balanced constitution

OBJECTIVE:To evaluate the extent of vascular endothelial dysfunction and preliminary identify serum protein biomarkers associated with obese individuals at risk for cardiovascular disease(CVD).METHODS:Fifteen obese volunteers with the phlegmdampness constitution or balanced constitution were recruited for this study respectively.The clinical baseline data was collected,and the vascular endothelial function was evaluated using the EndoPAT?.Blood samples were collected for the serum proteome analysis.The differences in the serum protein expression levels between the two groups were detected and the protein interaction network analysis,correlation analysis,receiver operating characteristic(ROC)curve analysis,and random forest model investigation were conducted.RESULTS:There were no statistical differences found in the baseline data.For vascular endothelial function,the reactive hyperemia index(RHI)of the phlegm-dampness constitution obese group was significantly lower than that of the balanced constitution obese group(1.46±0.30 vs 2.82±0.78,P<0.0001),indicating vascular endothelial dysfunction.There are 66 differentially expressed serum proteins between the two groups.apolipoprotein A2(Apo A2),angiotensin-converting enzyme 2(ACE-2),interleukin-33(IL-33),and forkhead box P3(FoxP3)showed significant differences and area under curve values of their ROC curves were greater than 0.7 and correlated significantly with RHI.CONCLUSION:Vascular endothelial dysfunction was present in the phlegm-dampness constitution obese group.Thus,alterations in the expression levels of key serum proteins,including Apo A2,ACE-2,IL-33,and Fox P3 could serve as potential biomarkers in the obese population at risk of CVD.

Effect of Electro-acupuncture on Expression of IRS-1/PI3K/GLUT4 Pathway in Ovarian Granulosa Cells of Infertile Patients with Polycystic Ovary Syndrome-Insulin Resistance of Phlegm-Dampness Syndrome

Objective:To evaluate the effect of electro-acupuncture(EA)in infertile patients with phlegmdampness polycystic ovary syndrome-insulin resistance(PCOS-IR).Methods:Seventy-six PCOS-IR patients who underwnet in vitro fertilization and embryo transfer(IVF-ET)were equally assigned to two groups according to a random digital table:the EA group and the control group,with 38 cases in each group.Before undergoing IVF,the two groups were treated with EA or pseudo-acupuncture,respectively,for 3 menstrual cycles.The intervention was 25 min twice a week until the day of oocyte collection.The selected acupoints were Zhongwan(RN 12),Tianshu(ST 25),Daheng(SP 15),Daimai(GB 26),Qihai(CV 6),Guanyuan(CV 4),and bilateral points including Xuehai(SP 10),Fenglong(ST 40),Zusanli(ST 36),and Yinlingquan(SP 9).Evaluation of phlegm-dampness syndrome score and IR score were carried out before and after treatment.Additionally,the number of oocytes retrieved,transplantable embryo rate,high-quality embryo rate,clinical pregnancy rate and live birth rate were compared between the two groups.Real-time polymerase chain reaction analysis was used to monitor the m RNA expression of the insulin receptor substrate(IRS-1),phosphatidylinositiol 3-kinase(PI3 K)and glucose transport factor 4(GLUT4)in ovarian granulosa cells.Results:EA treatment reduced the phlegm-dampness syndrome score as well as the IR scores compared with the control group(P<0.05).No significant differences in the number of oocytes retrieved and clinical pregnancy rate between the two groups(P>0.05).Moreover,the transplantable embryo rate[49.0%(284/580)vs.41.9%(273/652)],high-quality embryo rate[36.6%(104/284)vs.27.8%(76/273)],and live birth rate[50%(19/38)vs.26.3%(10/38)]in the EA group were significantly higher than in the control group(P<0.05).Gene expression analyses revealed significantly elevated IRS-1,PI3 K and GLUT4 m RNA in ovarian granulosa cells of the EA group compared with the control group(P<0.05).Conclusions:EA may ameliorate the effects of phlegm-dampness syndrome and ovarian IR in PCOS-IR patients.Mechanistically,this effect might be through an upregulation of the IRS-1/PI3 K/GLUT4 signaling pathway,which may result in improved oocyte quality and embryonic development potential.(Registration No.Chi CTR1800015453)

Exploration of the Relationship between Phlegm-Dampness Constitution and Polymorphism of Low Density Lipoprotein Receptor Genes Pvu Ⅱand AvaⅡ

Objective： To explore the polymorphism of low density lipoprotein receptor （LDL-R） genes Pvu Ⅱ end Ave Ⅱ in e population with phlegm-dampness constitution （PDC）. Methods： Polymorphism of LDL-R genes at Pvu Ⅱ end Ave Ⅱ of 48 persons with gentle constitution （GC） end 61 with PDC were analyzed with PCR-RELP technique, end their serum contents of lipids end glucose were determined end compared as well. Results： The A＋ ellelic end P- ellelic frequency were higher end the P＋ ellelic frequency was lower in subjects with PDC then those in subjects with GC, which were 0.3083 vs 0.1771, 0.9098 vs 0.7708 end 0.0902 vs 0.2292, respectively, ell showing significant difference between the two groups （P〈0.05）. Comparison of the two groups in serum levels of triglyceride （TG）, fasting blood glucose, 2 h postprandial blood glucose, end 2 h postprandial insulin showed that ell the parameters were higher in subjects with PDC then in subjects with GC respectively, showing significant difference （P〈0.05）. Conclusion： PDC is related with the P- end A＋ ellelic frequency of higher LDL-R genes at Pvu Ⅱ end Ave Ⅱ, therefore, the polymorphism of LDL-R genes could be taken as one of the genetic markers for PDC, end humans with PDC ere more liable to suffer from blood lipids end glucose disorder then those with GC.

Optimal exercise intensity and volume to impact rats with Traditional Chinese Medicine phlegm-dampness constitution

Objective:We aim to investigate the effects of different exercise intensities and volumes on Phlegm-dampness constitution(PDC).Methods:The rats were fed with high-fat food and lived in 75%–85%humidity for 6 weeks to establish the model of PDC.Then PDC rats were screened and intervened by varying exercise intensities for 8 weeks.Weight,constitution scores,blood and liver tissues were collected to detect the concentration of serum total cholesterol(Tch),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),homocysteine(Hcy),blood glucose(GLU),alanine transaminase(ALT)and percentage of lipid droplet area in liver tissue(PLDA).Results:The weight,Tch,TG,HDL-C,LDL-C,Hcy,GLU,and ALT of rats in moderate-intensity exercise group returned to normal.The rats with high-intensity or low-intensity exercise did not recover as well as moderate-intensity.Conclusion:Different exercise intensities and volumes have different effects on PDC,moderate-intensity exercise over an 8-week intervention is most appropriate.

Study on the Resolving Phlegm Effect of D-Limonene in Mice with Spleen Deficiency and Phlegm-Dampness Syndrome

Objective:According to Traditional Chinese Medicine theory,spleen deficiency and phlegm-dampness syndrome(SDPDS)are caused by abnormal water metabolism in the body because of spleen dysfunction.The purpose of this article was to evaluate the efficacy of D-limonene(DL)in resolving phlegm in mice with SDPDS from the perspective of regulating the level of aquaporin 3(AQP3).Methods:The model of SDPDS was induced in mice using the multifactor modeling method,which combines internal and external dampness.An artificial climate box was used to create a humid environment,whereas the irregular diet was caused by different feeding methods on odd-even days.The mice were divided into blank control,model group,DL low-dose,DL high-dose,and positive groups.The mice were modeled and treated for 7 day.Levels of gastrin and amylase(AMS)in the serum,mucus secretion in the trachea,and AQP3 in the tissue near the gastric cardia.Results:DL significantly reduced mucus secretion in the trachea(P<0.001).It also increased the level of AMS in the serum(P<0.01)and decreased the level of AQP3 in the gastric tissue(P<0.01).Conclusions:Mice with SDPDS exhibited disturbed water metabolism and significantly increased AQP3 levels.DL can restore the levels of AQP3 and plays an important role in resolving phlegm.This study may also help expand the efficacy of natural drugs containing DL and improve the utilization of natural drug resources.

Clinical and Experimental Study on Treatment of Fatty Liver of Phlegm-Dampness Stagnation Type by Zhiganle (脂肝乐) Capsule

Objective: To explore the mechanism of Zhiganle capsule (脂肝乐, ZGLC) in treating fattyliver of Phlegm-Dampness Stagnation type. Methods: One hundred and forty-six patients were treated withZGLC, the effects were observed and compared with those treated with Dongbao Gantai (DBGT, 东宝肝泰)tablets as control. The effect of ZGLC on ethionine induced hyperlipemia model rats was also observed. Results:The effect of ZGLC was obviously better than those of DBGT in relieving or removing symptoms, normalizingthe ultrasonic abnormal findings, lowering the serum levels of lipid and enzyme and improving hemorrheologicfunctions. Experimental study showed that ZGLC could reduce the blood lipids, inhibit the lipid deposition inliver. As compared with the model group, level of triglyceride in the ZGLC treatment group were much lowered(P < 0. of ) and the effect was better than those in the control group. Conclusions: ZGLC has the effect of reducing blood triglyceride level, inhibiting lipid deposition in liver and improving hemorrheologic function.Keywods: Zhiganle capsule; Phlegm--Dampness Stagnation; animal model of fatty liver

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Clinical Effect of Yinma Zhisou Decoction Combined with Western Medicine in the Treatment of Acute COPD(Stagnation of PhlegmDampness in Lung)

[Objectives]To explore the effectiveness and safety of Yinma Zhisou Decoction in the treatment of AECOPD with phlegm-heat obstructing lung syndrome,and observe the symptoms of Yinma Zhisou decoction in the treatment of AECOPD patients with phlegm-heat stagnation lung syndrome improvement,inflammatory indicators,lung function,blood gas analysis,etc.[Methods]A total of 92 cases who met the inclusion criteria and exclusion criteria were included in the study according to 1∶1 were randomly divided into treatment group and control group,control group with conventional Western medicine treatment,the treatment group on the basis of control group with Yinma Zhisou decoction treatment,a course of 10 d.The changes in TCM syndrome score,arterial blood gas analysis,pulmonary function,serum inflammatory indicator,mMRC score,and the CAT scores between the two groups after treatment were observed.All collected data were statistically analyzed with SPSS 26.0.[Results](i)Comparison of the curative effect between the two groups.After treatment,the total effective rate of the treatment group was 93.5%,which was higher than that of the control group(76.1%,P<0.05).The curative effect of the treatment group was better than that of the control group(P<0.05).(ii)Comparison of single TCM symptom scores.After treatment,the single TCM symptom scores of the two groups were lower than before(P<0.05),but the improvement of wheezing,cough,phlegm and chest pain in the treatment group was better than that in the control group(P<0.05).There was no significant difference in the improvement of wheezing sound(P>0.05),but there was a trend of difference.(iii)Comparison of mMRC score and CAT score between the two groups.After treatment,the mMRC score and CAT score of the two groups were significantly improved(P<0.05),and the difference was statistically significant.(iv)Comparison of blood gas analysis indicators between the two groups.Intra-group comparison after treatment:PaOof the control group and the treatment group after treatment was significantly improved compared with that before treatment(P<0.05),and the difference was statistically significant.Comparison between the two groups after treatment:PaOand PaCOof the two groups after treatment(P>0.05),and the difference was not statistically significant.It indicated that Yinma Zhisou decoction had more advantages in improving hypoxia.(v)Comparison of pulmonary function between the two groups.After treatment,the FEV1/FVC of the treatment group and the control group was significantly different(P<0.05),and there was a statistical difference(P<0.05).(vi)Comparison of inflammatory indicators between the two groups.Intra-group comparison after treatment:the inflammatory indicators WBC,CRP and PCT in the control group and the treatment group were significantly lower than before treatment(P<0.05),and the differences were statistically significant.It indicated that Yinma Zhisou decoction could effectively control inflammation and reduce inflammation reaction.[Conclusions]Compared with the control group,Yinma Zhisou decoction has obvious advantages in improving cough,expectoration and wheezing,and can significantly improve the total effective rate.Compared with Western medicine alone,Yinma Zhisou decoction can significantly improve patients’hypoxia,reduce mMRC and CAT scores,improve body activity tolerance,improve airway symptoms,and significantly reduce WBC,CRP,PCT and other serum inflammatory indicators.It can be seen that Yinma Zhisou decoction combined with Western medicine treatment is obviously better than simple Western medicine treatment,the curative effect is better,and the safety is better,and there are no serious adverse reactions in the course of the trial.

Correlation between tobacco and alcohol use and Traditional Chinese Medicine constitutions: an analysis based on a sample from general population of China

OBJECTIVE:To investigate the relationship between smoking and drinking and constitution types from Traditional Chinese Medicine(TCM)theory.METHODS:The Constitution in Chinese Medicine Questionnaire(CCMQ)was used.A sample of 8448subjects from nine provinces and municipalities was chosen from the database of the TCM constitution and health status survey.Constituent ratio and correspondence analysis were used to find the relationship between smoking and drinking and TCM constitution.RESULTS:There was an association between smokinganddrinkingandtheGentleness,phlegm-dampness,and dampness-heat type constitutions.People that did not smoke tobacco or drink alcohol tended to have Yang-deficiency,Yin-deficiency,Qi-depression,Qi-deficiency,special diathesis,orblood-stasis type constitutions.CONCLUSION:There was a significant correlation between tobacco and alcohol use and TCM constitutions.To improve patient health,more attention should be paid to tobacco and alcohol control.