Composition of intestinal microflora associated with Yang-deficiency

Objective:In this study,we investigated the composition of the intestinal microflora associated with Yang-deficiency and the function-specific members of this microflora.Method:Subjects with normal and Yang-deficient constitutions were recruited in Beijing,with 30 subjects in each group.Illumina high-throughput sequencing was used to sequence the DNA of their fecal bacteria in the 16S rRNA V3-V4 region.The species abundance and distribution of the intestinal microflora in each specimen were determined with a statistical analysis of these sequences.We identified an underlying taxonomic trend with nonparametric PCoA and other statistical techniques.Results:(1) Subjects with Yang-deficiency displayed more uneven abundances of the taxa in their intestinal microflora than the subjects with normal Yang;(2) several differences were observed in the proportions of certain bacterial groups between the Yang-deficient and control groups;and (3) the fallowing bacteria differed significantly between the two groups:(i) Brevundimonas,Leuconostoc,Turicibacter,Defluviitaleaceae incertae sedis,Rothia,and Butyricimonas were slightly higher in the Yang-deficient subjects,whereas (ii)Megasphaera,Ruminococcus,and Lachnospira were higher in the normal subjects than in the Yang-deficient subjects.

Serum protein profile of yang-deficiency constitution in traditional Chinese medicine revealed by protein microarray analyses

Background:Based on the theory of traditional Chinese medicine (TCM) and preepidemiological investigation,Professor Qi Wang classified the entire human population into nine constitutions and put forward the theory of 'Nine-Constitution Medicine.' Among these constitutions,the main feature of the yang-deficiency constitution (YADC) is intolerance of the cold,which has been proven to reduce quality of life and confer susceptibility to specific diseases.Previous studies explored the genetic and transcriptional bases of YADC.In this experiment,we explored the potential mechanism of YADC using protein microarray,to deepen our understanding of its biological mechanism.Methods:Subjects identified with a YADC (n =12) or a balance constitution (BC;n =12) in accordance with the Classification and Determination Standards of Constitutions in Traditional Chinese Medicine were selected.Blood was collected to separate serum and protein microarray technology was used to analyze serum protein expression.Results:The clustering of subjects' constitutions based on protein expression profiling largely coincided with the TCM classification.Based on false discovery rate correction (P <.01) and fold change ≥ 5 or ≤ 0.2,a total of 85 proteins differentially expressed in YADC compared with their status in BC were selected,including 64 upregulated and 21 downregulated ones.Enrichment analysis suggested that subjects with YADC are susceptible to endocrine and energy metabolism disorders,as well as decline in immune function.Conclusion:This study revealed that YADC exhibits systematic differences in its protein expression profile.Moreover,we can potentially explain the characteristics of YADC partly via differentially expressed proteins.

Progress in the study of Yang-deficiency constitution in terms of Traditional Chinese Medicine:a narrative review

The Yang-deficiency constitution has been a hot topic of interest in recent years.Important research progress has been made in quantitative diagnostic criteria and auxiliary identification technologies,modern biological interpretations of the constitution characteristics,the association between the Yang-deficiency constitution and diseases,as well as the mechanisms of regulating the constitution.Yet,there are still some deficiencies and limitations.Herein,we systematically reviewed the research progress on the Yang-deficiency constitution by searching articles published in China National Knowledge Infrastructure Database,China Science and Technology Journal Database,Wanfang Database,PubMed,and Embase.The future development direction of the Yangdeficiency constitution should focus on developing objective constitution identification techniques,establishing animal models of Yang-deficiency constitution,conducting constitutional and diseaserelated research with a high level of evidence,combined with research on the full chain of the nine constitutions so as to obtain more scientific and objective conclusions.

Genome-wide Association Study on Susceptibility Genes Associated with Yang-Deficiency Constitution:A Small Sample Case-Control Study

Objective： To explore susceptibility genes associated with yang-deficiency constitution using single nucleotide polymorphism （SNP） genotyping. Methods： Based on an epidemiological survey, 30 volunteers with yang-deficiency constitution and 30 volunteers with a balanced constitution were included according to the Classification and Determination Standards of Constitutions in Traditional Chinese Medicine. Peripheral blood was collected and DNA was extracted from white blood cells. A genome-wide association study （GWAS） was conducted by SNP 6.0 genotyping at the Beijing Capital Bio Corporation Ltd. A minimum association P-value （Fisher＇s exact value） of less than 104 in the allele, genotype, dominant, and recessive models served as the standard for significant association of SNP with yang-deficiency constitution. Results： Among the four genetic models, a total of 42 SNPs were significantly associated with yang-deficiency constitution （Fisher＇s exact P-values 〈10^4）. These SNPs were adjacent to more than 20 genes, including RGS6, mGluR5, GAPDHL19, and II〈ZF1. Conclusions： Yang-deficiency constitution exhibits the characteristics of polygenic inheritance. This pilot study suggests that the polymorphisms in RGS6, mGluR5, GAPDHL19, and IKZF1 are associated with changes in cyclic adenosine monophosphate and cyclic guanosine monophosphate levels, memory, metabolic energy status, and immune function, respectively in people with yang-deficiency constitution.

Consistency between traditional Chinese medicine constitution-based classification and genetic classification

Background:We studied the consistency between two classification systems for categorizing patients:traditional Chinese medicine(TCM)constitution-based methods,versus genetic clustering.Genetic classification in constitutional identification was also evaluated.Methods:A TCM physician evaluated the constitution of each patient,according to four examinations(inspection,auscultation-olfaction,interrogation,and palpation).Those who met the criteria for Yang-deficient,Yin-deficient,and balanced constitutions were enrolled in the study.Peripheral blood samples were obtained from the participants,and peripheral blood mononuclear cells were separated from the samples within 2 hours.Total RNA extraction from the white blood cells was performed;and an Affymetrix HG-U133 Plus2.0 array was used to determine the peripheral blood gene expression profiles.The samples were classified using a support vector machine genetic classifier,and the“leave-one-out”method was used for validation.Results:The global gene expression profiles of 32 samples were grouped into three categories,and the samples in each of the gene categories corresponded with the three constitution categories.The three constitution types were distinguished using the genetic classifier with 165 genes.The accuracy of the prediction classification was greater than 95%using mathematical method.Conclusions:Participants with Yin-deficient,Yang-deficient,and balanced constitutions have varying physical characteristics and gene expression patterns.Additionally,the results from TCM constitution classification matched those obtained by genetic classification.Finally,our preliminary gene classifier distinguishes among Yin-deficient,Yang-deficient,and balanced constitutions,and provides a methodological basis for identifying the different constitutions.

Study of Differential Serum Metabolites in Patients with Adenomatous Polyps of Colon and Yang-Deficiency Constitution Based on Ultra-Performance Liquid Chromatography-Mass Spectrometry

Objective:To study the differences between the serum metabolites in patients with adenomatous polyps of the colon and yang-deficiency constitution and those without colon polyps and with balanced constitution,and look for biomarkers that can be used to distinguish between the two groups.Methods:General patient information was gathered,and Chinese medicine constitutions were collected in 940 patients who underwent electronic colonoscopy.A total of 119 patients with adenomatous polyps of the colon and yang-deficiency constitution were included in the experimental group,and 150 patients without colon polyps and with balanced constitution were included in the control group.Metabolomics analysis was performed on the fasting venous blood obtained from each patient in both groups.Principal component analysis and orthogonal partial least squares discriminant analysis were performed on the detection results,potential biomarkers were screened,metabolic pathway changes were determined,and the metabolic processes involved were discussed.Results:A total of 59 differential biomarkers between the experimental group and the control group were identified.The differential metabolites were found mainly in the glycerophospholipid metabolism pathway,and the bile acid 3-oxo-4,6-choladienoic acid was the biomarker that distinguished the experimental group from the control group.Conclusions:With the help of metabolomics analysis,the differential metabolites in patients with adenomatous polyps of the colon and yang-deficiency constitution and those in patients without colon polyps and with balanced constitution could be identified.The biomarker 3-oxo-4,6-choladienoic acid may have potential diagnostic value in patients with adenomatous polyp of the colon and yang-deficiency constitution.(Trial Registration No.NCT02986308)

Moxibustion plus acupuncture improves the efficacy and quality of life of patients with polycystic ovary syndrome: A randomized controlled trial

Objective:To observe the effect of moxibustion combined with acupuncture on the efficacy and quality of life of spleen-kidney-yang-deficiency patients with polycystic ovary syndrome(PCOS).Materials and Methods:A total of 90 patients with PCOS of spleen-kidney yang deficiency type from January 2020 to March 2021 were selected and randomly divided into three groups,30 patients each in the moxibustion group,acupuncture group,and moxibustion combined with acupuncture group(combined group).The changes of body mass index(BMI),waist-to-hip ratio(WHR),fasting insulin(FINS),fasting plasma glucose(FPG),homeostasis model assessment-insulin resistance index(HOMA-IR),follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),triglycerides(TG),total cholesterol(TC),traditional Chinese medicine(TCM)clinical symptom score,efficacy,and quality of life score in the three groups were observed before and 3 months after treatment.Results:After treatment,(1)compared with the moxibustion and acupuncture groups,the combined group was more effective in reducing BMI(F=5.274,P<0.05)and WHR(F=8.246,P<0.05)in PCOS patients;(2)FINS,FPG,and HOMA-IR were reduced in all three groups,and the reduction in these three indicators was more pronounced in combined group(FINS:F=5.637,P<0.05,FPG:F=4.915,P<0.05,HOMA-IR:F=3.817,P<0.05);(3)The FSH,LH,and T levels of patients in the combined group were better than those in the remaining two groups(FSH:F=4,163,P<0.05,LH:F=5.098,P<0.05,T:F=7.038,P<0.05);(4)The TG and TC of patients in the combined group were lower than those in the remaining two groups(TG:F=4.806,P<0.05,TC:F=3.828,P<0.05);(5)The TCM clinical symptom score of the combined group was significantly lower than those of the moxibustion and acupuncture groups(F=4.547,P<0.05);(6)The quality of life of patients in the combined group improved more significantly than the moxibustion and acupuncture groups(F=6.239,P<0.05);(7)The total effective rate of the combined group was higher than that of the moxibustion and acupuncture groups(χ2=6.947,P=0.031).Conclusion:The combination of moxibustion and acupuncture can effectively reduce the BMI and WHR of PCOS patients,improve the endocrine function,sex hormone level,TCM symptoms and quality of life of patients,and has significant efficacy and high safety,which is worthy of wide clinical application.

On Glucocorticoid receptors of Peripheral Leucocytes in Patiente with Yang-Deficiency

The leucocytic glucocorticoid receptors (GCR) were estimated in 20 patients with Yang-Defi-ciency Syndrome using a radioligand binding method and plasrna cortisol was measured at the same time. Re-sults showed that the levels of plasma cortisol and leucocytic GCR of patients were all significantly lower thanthe healthy control group, and the changes were more significant in 10 cases of hypofunction of endocrinopa-thy with manifestation of Yang-Deficiency and Yang-Doficiency patients aged < 40 years. The mechanism andrelationship between GCR and plasma cortisol were also discussed in this article.

Spleen-kidney supplementing formula alleviates insulin resistance via regulating AKT/glycogen synthase kinase 3β pathway in rats with type 2 diabetic induced by high-fat diet

OBJECTIVE: To explore the molecular mechanism underpinning the action by investigating its effect on glycogen content and AKT(also known as protein kinase B)/glycogen synthase kinase 3β(GSK-3β) pathway in the liver of rats with type 2 diabetic induced by high-fat diet.METHODS: The rat model of type 2 diabetes was induced by high-fat diet and multiple low-dose streptozotocin injection. Diabetic rats were divided into five groups: the model control group, the Metformin group, spleen-kidney supplementing formula groups of low, medium and high doses. Fasting blood glucose(FBG) levels were measured before treatment and every two weeks during treatment.After the treatment, oral glucose tolerance test was performed, and hemoglobin A1c (HbA1c) and C-peptide were measured to assess the formula's effect on glucose metabolism and insulin resistance. The protein expression levels of AKT, GSK-3βand their phosphorylated forms in the liver were also measured to study the formula's role in insulin signaling pathway.RESULTS: Spleen-kidney supplementing formula significantly relieved the symptoms of polydipsia,polyuria and weight loss in type 2 diabetic rats, reduced FBG and HbA1c levels, increased glycogen content, and improved insulin sensitivity. The anti-diabetic effects of spleen-kidney supplementing formula are dose dependent. It also increased the total AKT protein level and the GSK-3β phosphorylation in the liver of type 2 diabetic rats.CONCLUSION: Spleen-kidney supplementing formula has hypoglycemic effect and relieves insulin resistance by enhancing AKT/GSK-3β signaling pathway in the liver of type 2 diabetic rats.

Observation on Clinical Effects of Tuina plus Hot Compress for Infantile Diarrhea Due to Spleen-kidney Yang Deficiency

Objective： To observe the clinical effects of tuina plus hot compress for nfantile diarrhea clue to spleen-kidney yang deficiency. Methods： One hundred cases were randomly divided into a treatment group and a control group, 50 cases in each group. The treatment group was treated with tuina plus hot compress. The control group was treated with oral administration of Live Combined Bifidobacterium, Lactobacillus and Enterococcus powder （Bifico）. The clinical effects were observed after 5-day intervention. Results： The total effective rate was 94.0% in the treatment group and was 66.0% in the control group. The difference in the total effective rate between the two groups was statistically significant （P〈0.05）. The clinical effect was better in the treatment group than that in the control group. Conclusion： Tuina plus hot compress for infantile diarrhea due to spleen-kidney yang deficiency can produce a better clinical effect than oral administration of Bifico.

Observation on effects of moxibustion at abdominal acupoints for slow transit constipation due to yang deficiency of the spleen and kidney

Objective： To observe the clinical effects of moxibustion at abdominal acupoints for slow transit constipation（STC） due to yang deficiency of the spleen and kidney. Methods： A total of 52 cases with slow transit constipation in conformity with the inclusion criteria were selected and divided into a control group and an observation group according to their visit order and random digital table, 26 cases in each group. Patients in the control group received routine nursing guide. Besides the same routine nursing guide, patients in the observation group received moxibustion at the abdominal acupoints, once every day. The course of the treatment was 4 weeks in the two groups, and the 3-month follow-up was given after the course was finished, for comparing the clinical symptoms, results of colon transit tests, scores of depression/anxiety scale and nursing satisfaction. Results： The total effective rate was 92.3% in the observation group and 69.2% in the control group, with a significant difference between the two groups. After the treatment and during the follow-up checks, the scores of Chinese medical symptoms in the two groups were remarkably decreased than those before the treatment（all P〈0.01）; the scores of the observation group were obviously lower than those in the control group（all P〈0.01）. The discharge rates of the markers in the two groups were remarkably increased than those at the same time period before the treatment; moreover, the discharge rates of the markers at various time periods were remarkably better in the observation group than those in the control group（P〈0.01）. SDS and SAS scores were remarkably decreased after the treatment in the two groups（P〈0.01）. In comparison of SDS score between the two groups after the treatment, the difference was statistically significant（P〈0.01）. In comparison of SAS score between the two groups after the treatment, the difference was statistically significant（P〈0.05）. The nursing satisfaction was 96.2% in the observation group after the treatment, obviously better than that in the control group（73.1%）. The recurrence rate was 8.3% in the observation group, remarkably lower than that in the control group（33.3%）. Conclusion： Moxibustion at the abdominal acupoints plus routine nursing can remarkably improve the colon transit functions and anxious and depressive emotion in patients with STC, and the therapeutic effects are remarkable. Not only the clinical satisfaction is higher, but the recurrence rate is obviously lower than that of routine nursing.