Metabolomic profiling and alleviation of oxidative stress of Huangjing Zanyu capsule treating oligoasthenospermia.

Background: The traditional Chinese medicine (TCM) patent medicine Huangjing Zanyu capsule (HJZY capsule) has achieved satisfactory clinical effects in the treatment of oligoasthenospermia (OAS). This study aimed to elucidate the impact of HJZY capsule on the reproductive system, focusing on oxidative stress and metabolism profiling during the intervention, to clarify the therapeutic mechanism of HJZY capsule in treating OAS. Methods: Cyclophosphamide was used to establish OAS model rats. Time-sequence specimen collection was applied to monitor the dynamic development of the pharmacological effect of HJZY capsule. Superoxide dismutase (SOD), glutathione peroxidase (GPX), and malonaldehyde (MDA) were evaluated by biochemistry kits to examine the impact of HJZY capsule on oxidative stress. Non-targeted metabolomics was conducted for urine and testis samples, respectively, to investigate metabolic pathways through which the HJZY capsule takes effect. Results: The HJZY capsule elevated sperm density from 62.1±8.28, passing 68.4±7.52, to 75.9±8.48×106/mL, and sperm motility from 62.0%±3.94%, passing 70.8%±9.72%, to 68.8%±4.37%. Meanwhile, SOD (P<0.05 in week 2) and GPX activity levels of HJZY groups were elevated to a certain degree, respectively, and lipid oxidation was attenuated, as shown by decreased MDA content (P<0.05 in week 2). Metabolomics results showed that the HJZY capsule could modulate pathways including taurine metabolism, purine and pyrimidine metabolism, glycerolipid and glycerophospholipid metabolism, and multiple amino acid metabolisms, among others. The cluster analysis results showed that urinary and testicular metabolomics differed in the strength of discrimination between rats in the OAS model and the HJZY groups. Conclusions: The HJZY capsule exerts a comprehensive effect on OAS through influencing various metabolic pathways. Non-targeted metabolomics provides an effective way for profiling complex TCM prescriptions.

Traditional Chinese Medicine Constitution Identification Based on Objective Facial and Tongue Features: A Delphi Study and a Diagnostic Nomogram for Blood Stasis Constitution.

Objective: The aim of this study was to systematically summarize and form an expert consensus on the theoretical experience of tongue and facial features for the identification of nine types of traditional Chinese medicine (TCM) constitution. Additionally, we sought to explore the feasibility of TCM constitution identification through objective tongue and facial features. Methods: We used Delphi method to investigate the opinions of experts on facial and tongue feature items for identifying TCM constitution. We developed and validated a diagnostic nomogram for blood stasis constitution (BSC) based on objective facial and tongue features to demonstrate the reliability of expert consultation. Results: Eleven experts participated in two rounds of expert consultation. The recovery rates of the two rounds of expert consultation were 100.0% and 90.9%. After the first round, 39 items were screened out from 147 initial items, and 2 items were supplemented by experts. In the second round, 7 items were eliminated, leaving 34 items for 8 types of TCM constitution. The coefficient of variation in the first round was 0.11-0.49 for the 147 items and 0.11-0.29 for the included items. The coefficient of variation in the second round was 0.10-0.27 for the 41 items and 0.10-0.20 for the included items. The W value was 0.548 (P < 0.001) in the first round and 0.240 (P < 0.001) in the second round. Based on expert consultation, we selected BSC as an example and developed and validated a diagnostic nomogram consisting of six indicators: sex, hair volume, lip color-dark purple, susceptibility-facial pigmentation/chloasma/ecchymosis, zygomatic texture-red blood streaks, and sublingual vein-varicose and dark purple. The nomogram showed good discrimination (AUC: 0.917 [95% confidence interval [CI], 0.877-0.956] for the primary dataset, 0.902 [95% CI, 0.828-0.976] for the validation dataset) and good calibration. Decision curve analysis demonstrated that the nomogram was clinically useful. Conclusion: This is the first study to systematically summarize the existing knowledge and clinical experience to form an expert consensus on the tongue and facial features of nine types of TCM constitution. Our results will provide important prior knowledge and expert experience for future constitution identification research. Based on expert consultation, this study presents a nomogram for BSC that incorporates objective facial and tongue features, which can be conveniently used to facilitate the individualized identification of BSC.

Development and evaluation of short form of constitution in Chinese medicine questionnaire: a national epidemiological survey data of 21 948 case.

OBJECTIVE: To develop the best short form of constitution in Chinese medicine questionnaire (CCMQ) and evaluate its psychometric properties in Chinese population. METHODS: A total of 21 948 subjects were used to refine the short form. Correlation coefficient, exploratory factor analysis (EFA) and Cronbach's alpha coefficient were used to analyze and select items to form the short form. Separate sample of 205 subjects were collected to further evaluate the short from. EFA, confirmatory factor analysis (CFA), item-scale correlation, discriminant validity, internal consistency reliability and split-half reliability were carried out to evaluate the short form. RESULTS: The short form CCMQ included 26 items. Seven common factors of characteristic root > 1 were extracted to explain 58.488% of the total variation. Result of CFA was consistent with the 9-factors structure. The mean differences of Blood-stasis body constitution and Qi-stagnation body constitution had statistical significance in body mass index differentiation. Cronbach's alpha coefficient of short form CCMQ was 0.863. The split-half reliability of total scale was 0.813, and each scale was 0.568-0.770. The item-scale correlations ranged from 0.620-0.849. CONCLUSION: The short form CCMQ consisted of 26 items with good psychometric properties. The short form should be recommended for the measurement of health of Chinese population in any clinical trial.

DIA-PRM Proteomic Analysis of Phlegm-Dampness Constitution with Glucolipid Metabolic Disorders by the Intervention of Hua Tan Qu Shi Recipe.

Background: Phlegm-dampness constitution as one of nine constitutions in traditional Chinese medicine (TCM) has been a high risk factor for glucolipid metabolic disorders (GLMD). Based on our previous findings, Hua Tan Qu Shi recipe (HTQSR) could effectively improve metabolic indicators of GLMD by targeting on phlegm-dampness constitution. However, the proteomic mechanisms of GLMD with the treatment of HTQSR targeting on phlegm-dampness constitution remain unknown. Methods: Clinical participants from phlegm-dampness constitution with the prediabetic state (T), phlegm-dampness constitution with marginally elevated blood lipids (Z), and phlegm-dampness constitution before sickness (W) were included in this study, who orally took HTQSR for 12 weeks and, respectively, marked AT, AZ, and AW. Data-independent acquisition (DIA) and parallel reaction monitoring (PRM) were performed to identify the differential proteins; then, Venn analysis was used to investigate coexpressed and coregulated proteins. In addition, ingenuity pathway analysis (IPA) software was utilized to explore the related pathways and diseases and biofunctions. Results: LXR/RXR activation, acute phase response signaling, and production of nitric oxide and reactive oxygen species in macrophages were obviously activated between the T and AT groups, as well as the Z and AZ groups. In contrast, these three pathways were inhibited between the W and AW groups. Importantly, one coexpressed and coregulated differential protein, B2MG, was validated by PRM among all groups. Conclusions: This work firstly reported the underlying proteomic mechanisms of GLMD with the treatment of HTQSR targeting on phlegm-dampness constitution, indicating that intervention of phlegm-dampness constitution might be a novel strategy for the preventive treatment of GLMD.

Analyses of Long Noncoding RNA and mRNA Profiles in Subjects with the Phlegm-Dampness Constitution.

BACKGROUND: Constitution in traditional Chinese medicine (TCM) plays a key role in the genesis, development, and prognosis of diseases. Phlegm-dampness constitution (PDC) is one of the nine constitutions in TCM, susceptible to metabolic disorders, which is mainly manifested by profuse phlegm, loose abdomen, and greasy face. Epidemiologic, genomic, and epigenetic studies have been carried out in previous works, confirming that PDC represents a distinctive population with microcosmic changes related to metabolic disorders. However, whether long noncoding RNAs (lncRNAs) play a regulatory role in metabolic disease in subjects with PDC remains largely unknown. We aimed to investigate distinct lncRNA and mRNA expression signatures and lncRNA-mRNA regulatory networks in the phlegm-dampness constitution (PDC). METHODS: The peripheral blood mononuclear cells (PBMCs) were isolated from the subjects with PDC (n = 13) and balanced constitution (BC) (n = 9). The profiles of lncRNAs and mRNAs in PBMCs were analyzed using microarray and further validated with RT-qPCR. Subsequently, pathway analysis was performed to investigate the function of differentially expressed mRNAs by using Ingenuity Pathway Analysis (IPA). RESULTS: Results suggested that some mRNAs, which were regulated by the differentially expressed lncRNAs, were mainly enriched in lipid metabolism and immune inflammation-related pathways. This was consistent with the molecular characteristics of previous studies, indicating that the clinical characteristics of metabolic disorders in PDC might be regulated by lncRNAs. Furthermore, by making coexpression network construction as well as cis-regulated target gene analysis, several lncRNA-mRNA pairs with potential regulatory relationships were identified by bioinformatic analyses, including RP11-317J10.2-CA3, RP11-809C18.3-PIP4K2A, LINC0069-RFTN1, TTTY15-ARHGEF9, and AC135048.13-ORAI3. CONCLUSIONS: This study first revealed that the expression characteristics of lncRNAs/mRNAs may be potential biomarkers, indicating that the distinctive physical and clinical characteristics of PDC might be partially attributed to the specific expression signatures of lncRNAs/mRNAs.

Effect of Electroacupuncture on Gut Microbiota in Participants With Knee Osteoarthritis.

A close relationship between knee osteoarthritis (KOA) and gut microbiota has recently been described. Herein, we aim to investigate the effect of electroacupuncture (EA) on gut microbiota in participants with KOA. We conducted a study of 60 participants with KOA and 30 matched healthy controls (HCs). Sixty participants were allocated to either EA group (n=30) or sham acupuncture (SA) group (n=30). Five obligatory acupoints and three adjunct acupoints were punctured in the EA group. Eight non-acupoints that were separated from conventional acupoints or meridians were used for the SA group. Participants in both groups received 24 sessions within eight weeks. Fecal microbial analyses by 16S ribosomal RNA gene sequencing were carried out after collecting stools at T0 and T8 weeks (Four samples with changed defecation habits were excluded). The results showed that both Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) total score (P=0.043) and NRS score (P=0.002) decreased more in EA group than those in SA group. Moreover, EA could reverse more KOA-related bacteria including Bacteroides, [Eubacterium]_hallii_group, Agathobacter and Streptococcus. The number of significantly different genera between KOA patients and HCs were less after EA treatment than that after SA treatment. This meant that EA modified the composition of the gut microbiome, making it closer to healthy people, while not significantly affecting the microbial diversity. Two genera including Agathobacter (P=0.0163), Lachnoclostridium (P=0.0144) were statistically increased than baseline in EA group (paired Wilcoxon rank sum test). After EA treatment, Bacteroides (P=0.0394) was more abundant and Streptococcus (P=0.0306) was significantly reduced in patients who demonstrated adequate response than in those with inadequate response (Wilcoxon rank-sum test). Spearman correlation test between gut microbe and KOA clinical outcomes indicated that Bacteroides and Agathobacter was negatively correlated with NRS score, WOMAC total score, and WOMAC pain, stiffness and pain scores (P<0.001 or 0.05 or 0.01), while Streptococcus was positively correlated with them (P<0.05 or 0.01). Our study suggests that EA contributes to the improvement of KOA and gut microbiota could be a potential therapeutic target.

Effect of Knowledge/Practice of COVID-19 Prevention Measures on Return-to-Work Concerns; Attitudes About the Efficacy of Traditional Chinese Medicine: Survey on Supermarket Staff in Huanggang, China.

Objective: The objective of this study was to investigate how knowledge and practice of coronavirus disease 2019 (COVID-19) prevention measures affected concerns about returning to work among supermarket staff. Attitudes about the ability of traditional Chinese medicine (TCM) to prevent COVID-19 were also assessed. Methods: A cross-sectional study was conducted in Huanggang, Hubei Province, China from April 23 to 25, 2020. Participants were invited to fill out an electronic questionnaire on their cell phones. Results: The results showed that from 2,309 valid questionnaires, 61.5% of participants were concerned about resuming work. Major concerns included asymptomatic infection (85.01%) and employees gathering in the workplace (78.96%). Multivariate logistic regression indicated that the female gender, having school-aged children and pregnancy were risk factors for being concerned about resuming work, while good knowledge and practice of preventive measures were protective factors. Knowledge and practice of preventive measures were positively correlated. Among preventive measures, the highest percentage of participants knew about wearing masks and washing hands. Meanwhile, 65.8% of participants expressed confidence in the ability of TCM to prevent COVID-19, where 74 and 51.3% thought there was a need and a strong need, respectively, for preventive TCM-based products. Among them, 71.5% preferred oral granules. Regarding TCM as a COVID-19 preventative, most were interested in information about safety and efficacy. Conclusion: These findings suggested that promoting knowledge and practices regarding COVID-19 prevention can help alleviate concerns about returning to work. Meanwhile, TCM can feasibly be accepted to diversify COVID-19 prevention methods. Clinical Trial Registration:http://www.chictr.org.cn/, identifier: ChiCTR2000031955.

A Meta-Analysis of the Association between Diabetes Mellitus and Traditional Chinese Medicine Constitution.

OBJECTIVE: To explore the distribution of constitution types of diabetes mellitus (DM) in traditional Chinese medicine (TCM) and to provide evidence-based medicine basis for the prevention and treatment of diabetes. METHODS: PubMed, Embase, Web of Science, and three Chinese databases were searched to include research literature on the relationship between diabetes and TCM constitution. The single rate study of cross-sectional literature was conducted with RStudio software, and the control meta-analysis of the diabetic and nondiabetic population was performed with Review Manager 5.3 software. Two independent reviewers assessed the methodological quality of the studies' data. The main outcomes included the distribution of constitutional types in the diabetic population and the odds ratio (OR) between the two. Effect sizes are expressed as proportions or ORs with 95% confidence intervals (CI). RESULTS: A total of 28,781 diabetic cases were included in 87 articles. Yin-deficiency, phlegm-dampness, and qi-deficiency accounted for 18% (95% CI (15%, 20%), P < 0.01), 17% (95% CI (15%, 19%), P < 0.01), and 13% (95% CI (11%, 15%), P < 0.01) of the total diabetic cases. The risk of diabetes in people with yin-deficiency and phlegm-dampness was 3.06 (95% CI (1.38-6.78), P=0.006) and 1.89 (95%CI (1.05-3.42), P=0.03) times higher than that in those with other constitutions, respectively. The distribution of TCM constitution of DM patients varied significantly in different regions and ages. CONCLUSION: Yin-deficiency and phlegm-dampness are the common constitution types of diabetic people, and they may also be the risk factors of diabetes. Balanced constitution may be a protective factor of diabetes. More high-quality cohort and case-control studies need to be designed to provide more valuable evidence-based basis for assessing the correlation between DM and TCM constitution.

An Integrated Analysis of Network Pharmacology, Molecular Docking, and Experiment Validation to Explore the New Candidate Active Component and Mechanism of Cuscutae Semen-Mori Fructus Coupled-Herbs in Treating Oligoasthenozoospermia.

PURPOSE: One of the most common types of male infertility is recognized as oligoasthenozoospermia (OA), characterized by low sperm count and quality in males. As a traditional Chinese medicine (TCM), Cuscutae Semen-Mori Fructus coupled-herbs (CSMFCH) has been known to act a curative effect on OA for thousands of years. Nevertheless, the substantial basis and molecular mechanism of CSMFCH in treating OA remain elusive. METHODS: Herein, an integrated approach, including network pharmacology, molecular docking, and experiment validation, was utilized to reveal the new candidate active component and mechanism of CSMFCH in treating OA. RESULTS: The results show that kaempferol is the most significant bioactive component of CSMFCH on OA. The mechanism and targets of CSMFCH against OA are relevant to hormone regulation, oxidant stress, and reproductive promotion. In order to validate network pharmacology results, molecular docking and experiment validation were conducted. In detail, molecular docking was employed to verify the strong binding interactions between kaempferol and the core targets. UHPLC-Q-Orbitrap-MS was used to identify kaempferol in the CSMFCH extract. In vitro and in vivo experiments further proved CSMFCH and kaempferol could enhance the mouse Leydig (TM3) and mouse Sertoli (TM4) cell viability, improve the male reproductive organ weights, sperm quality, and decrease testis tissue damage in the OA mouse model induced by CP. CONCLUSION: Our results not only identify the new candidate active component of CSMFCH in treating OA but also provide new insights into the mechanisms of CSMFCH against OA.

Enlightenment about using TCM constitutions for individualized medicine and construction of Chinese-style precision medicine: research progress with TCM constitutions.

TCM constitution is a new branch of TCM. It provides enlightenment on individualized medicine, including the development of new models of individualized research based on nine constitutions, the acquisition of comprehensive health information for individuals, and establishment of a consistent individualized diagnosis and treatment system. Further, we propose a Chinese-style "precision medicine" based on individualization using the TCM constitutions.

COVID-19 Knowledge/Practices, Mental Status, and Return-To-Work Concerns Among Healthcare Workers in Huangzhou District, Huanggang City, China.

Objective: This study investigated the COVID-19-prevention knowledge and practices of healthcare workers (HCWs), their psychological states concerning the return to work, and their trust and requirements in using traditional Chinese medicine (TCM) to prevent and treat COVID-19. It is hoped that the study can serve as a reference for policy making during the resumption of work in other countries or regions experiencing similar situations. Methods: This study comprised a quantitative cross-sectional online survey design. Purposive sampling and Cluster sampling were used to recruit all HCWs working in public hospitals in Huangzhou District, Huanggang City, Hubei Province, China. From April 23 to May 14, 2020, surveys were sent electronically to all 13 public hospitals in this area. Results: In total, 2,079 responses were received and 2,050 completed forms were included. After analysis, 47.9 and 46.6% of HCWs indicated that they possessed very good knowledge or good knowledge of preventative measures, respectively. Multivariable log-binomial regression indicated that male, tertiary hospital, medical staff, and undergraduate/postgraduate qualification were associated with good knowledge. Good knowledge was also well-correlated with good practice (OR: 3.277; 95% CI: 2.734-3.928; P < 0.01). 59.8% of HCWs reported worries about resuming work; especially asymptomatic infections. The Self-Rating Anxiety Scale (SAS) indicated that 10.8% of participants had mild anxiety, 1.5% moderate anxiety, and 0.1% severe anxiety. Female, divorced/widowed, and working in a high risk hospital (the Huangzhou District People's Hospital was used for throat swab examinations of returning workers) were risk factors for concerns about resuming work and anxiety symptoms. However, good preventive knowledge was a protective factor for anxiety. HCWs' trust in using TCM to treat COVID-19 was significantly higher than their trust in using TCM for prevention (P < 0.001). Regarding preferences for preventative TCM products, oral TCM granules were the most preferred (62.4%). HCWs also indicated they wanted to know more about the clinical efficacy, applicable population, and adverse reactions of preventative TCM products (89.3, 81.1, and 81.4%, respectively). Conclusion: While HCWs had good knowledge of COVID-19 preventative measures, this did not eliminate the psychological impact of resumption of work. Promotion of COVID-19 prevention knowledge reduces the risk of infection, and alleviates the worries and anxiety symptoms of HCWs about resuming work (especially in administrative staff, those with low education, and those working in primary hospitals). Additional psychological support is required for female HCWs, divorced/widowed HCWs, and those working in high-risk hospitals. Finally, systematic trials of preventative TCM products are recommended.

[Medication experience of TCM Master Academician Wang Qi in the treatment of premature ejaculation: Data mining and analysis].

Objective: To analyze the academic thought, medication experience and prescription rules of Academician Wang Qi in the treatment of premature ejaculation (PE) using the TCM inheritance support platform (V2.5). METHODS: We collected and sorted out the medical records on the treatment of PE from Academician Wang Qi's Clinic. We established a database of medical records on the TCM inheritance support platform, analyzed the drugs and prescriptions in the database and explored new prescriptions using "statistical reports" and "data analysis" systems on the platform. RESULTS: A total of 91 effective prescriptions were recorded, involving 148 TCM drugs, with Phellodendron, Amomum Villosum, Polygala Tenuifolia, Tuckahoe, Lodestone, Oyster, Acanthopanax Senticosus, Uncaria, Tribulus, and Keel as the top 10 with the highest frequency of use, which were featured mainly by "warm" and "cold" concerning the four natures, "sweet", "bitter" and "pungent" relating to the five flavors, and acting on "kidney meridian", "liver meridian" and "heart meridian" in terms of the meridian tropisms. In addition, 5 new prescriptions were obtained through unsupervised entropy hierarchical clustering. CONCLUSIONS: In the treatment of PE, Academician Wang Qi employs tranquilizing the mind and consolidating the kidney (An Zhi Gu Shen) as the primary strategy, taking into account the three organs of heart, liver and kidneys, focusing on the phase of calming the mind or regulating the liver or clearing the kidney or controlling fire, and adding or reducing drugs according to different conditions and syndromes, which conforms to his diagnosis and treatment mode of "body differentiation－disease differentiation－syndrome differentiation". The analysis of the potential new prescriptions also accords with Academician Wang Qi's rules of medication, which can provide some ideas for the clinical treatment of and scientific researches on premature ejaculation in the future.

Clinical research linking Traditional Chinese Medicine constitution types with diseases: a literature review of 1639 observational studies.

OBJECTIVE: To analyze clinical studies on correlations between Traditional Chinese Medicine (TCM) body constitution types and diseases published in the past 10 years, and to provide an evidence base to support the use of such correlations for health maintenance and disease prevention. METHODS: We searched five databases for the period April 2009 to December 2019: China National Knowledge Infrastructure Database, Wanfang Database, China Science and Technology Journal Database, PubMed and Embase. Three types of observational studies on correlation between constitution types and diseases were included: cross-sectional, case-control and cohort studies. Descriptive statistical methods were employed for data analysis. RESULTS: A total of 1639 clinical studies were identified: 1452 (88.59%) cross-sectional studies, 115 (7.02%) case-control studies and 72 (4.39%) cohort studies covering 30 regions of China and five other countries (Malaysia, South Korea, Singapore, Thailand and France). The collection of studies comprised 19 disease categories and 333 different diseases. The 10 most commonly studied diseases were hypertension, diabetes, stroke, coronary atherosclerotic heart disease (CAHD), sleep disorders, neoplasm of the breast, dysmenorrhea, fatty liver disease, chronic viral hepatitis B and dyslipidemia. We found high distributions for each biased constitution type in different patient populations as follows: Qi-deficiency constitution in stroke, diabetes, chronic obstructive pulmonary disease, acquired immunodeficiency syndrome and hypertension; Yang-deficiency constitution in female infertility, osteoporosis, irritable bowel syndrome, gonarthrosis and dysmenorrhea; Yin-deficiency constitution in hypertension, diabetes, constipation, female climacteric states and osteoporosis; phlegm- dampness constitution in hypertension, stroke, fatty liver disease, diabetes and metabolic syndrome; damp-heat constitution in acne, chronic gastritis, chronic viral hepatitis B, human papillomavirus infection and hyperuricemia; blood-stasis constitution in CAHD, endometriosis and stroke; Qi-stagnation constitution in hyperplasia and neoplasms of the breast, insomnia, depression and thyroid nodules; and inherited-special constitution in asthma and allergic rhinitis. CONCLUSION: Eight biased TCM constitutions were closely related to specific diseases, and could be used to guide individualized prevention and treatment. More rigorously designed studies are recommended to further verify the constitution-disease relationship.

The Role of Chinese Medicine in Health Maintenance and Disease Prevention: Application of Constitution Theory.

Traditional Chinese medicine (TCM) has guided health maintenance and disease treatment for thousands of years and has been widely used in many countries around the world. TCM regards each individual as unique so all corresponding therapeutic and preventive approaches in TCM are personalized. Personalized medicine, also referred to as precision medicine, is an emerging medical paradigm that points toward a new direction for future medical development. TCM constitution studies the holistic body condition, which is affected by both inborn and acquired factors. Body constitution lays the foundation for disease diagnosis, prevention and treatment. Different constitution types predispose individuals to different disease susceptibilities. Examining an individual's unique body constitution can promote effective health management and benefit the application of personalized medicine significantly. This review will introduce and discuss the application of the TCM constitution for health maintenance and disease prevention. In last decade, a number of modern techniques have been employed in the constitution research to evaluate the health status of individuals. The TCM constitution reflects the current status and future trends of human health in four aspects, i.e., individual differences, life processes, psychological condition and adaptability to natural and social environments. This TCM constitution theory has already been applied in the Chinese public health management at different levels with promising outcome. The constitution theory and practice provide a new approach for health maintenance and disease prevention.

miRNA Expression Profile of Saliva in Subjects of Yang Deficiency Constitution and Yin Deficiency Constitution.

BACKGROUND/AIMS: Based on the theory of constitution in Traditional Chinese Medicine (TCM), the Chinese Han population has been classified into nine constitutions. Of these, Yang deficiency constitution mainly exhibit cold intolerance while Yin deficiency constitution mainly exhibit heat intolerance. Some studies have been carried out to explore the modern genetic and biological basis of such constitution classification, but more remains to be done. MicroRNA (miRNA) serves as post-transcriptional regulators of gene expression and may play a role in the classification process. Here, we examined miRNA expression profile of saliva to further improve the comprehensiveness of constitution classification. METHODS: Saliva was collected from Chinese Han individuals with Yang deficiency, Yin deficiency and Balanced constitutions (n=5 each), and miRNA expression profile was determined using the Human miRNA OneArray®v7. Based on 1.5 Fold change, means log2|Ratio|≥0.585 and P-value< 0.05, differentially expressed miRNA was screened. Target genes were predicted using DIANA-TarBasev7.0 and analysis of KEGG pathway was carried out using DIANA-mirPathv.3. RESULTS: We found that 81 and 98 differentially expressed miRNAs were screened in Yang deficiency and Yin deficiency constitution, respectively. Among them, 16 miRNAs were identical and the others were unique. In addition, the target genes that are regulated by the unique miRNAs were significantly enriched in 27 and 20 signaling pathways in Yang deficiency and Yin deficiency constitution, respectively. Thyroid hormone signaling pathway is present in both constitutions. These unique miRNAs that regulated target genes of thyroid hormone signaling pathway may be associated with cold intolerance or heat intolerance. CONCLUSION: The results of our study show that Yang deficiency and Yin deficiency constitutions exhibit systematic differences in miRNA expression profile. Moreover, the distinct characteristics of TCM constitution may be explained, in part, by differentially expressed miRNAs.

Effects of compound K, an enteric microbiome metabolite of ginseng, in the treatment of inflammation associated colon cancer.

Ginsenoside Rb1, a major component of different ginseng species, can be bioconverted into compound K by gut microbiota, and the latter possess much stronger cancer chemopreventive potential. However, while the initiation and progression of colorectal cancer is closely associated with gut inflammation, to date, the effects of compound K on inflammation-linked cancer chemoprevention have not been reported. In the present study, liquid chromatography quadrupole time-of-flight mass spectrometry analysis was applied to evaluate the biotransformation of Rb1 in American ginseng by human enteric microflora. The in vitro inhibitory effects of Rb1 and compound K were compared using the HCT-116 and HT-19 human colorectal cancer cell lines by a MTS assay. Cell cycle and cell apoptosis were assayed using flow cytometry. Using ELISA, the anti-inflammatory effects of Rb1 and compound K were compared for their inhibition of interleukin-8 secretion in HT-29 cells, induced by lipopolysaccharide. The results revealed that compound K is the major intestinal microbiome metabolite of Rb1. When compared with Rb1, compound K had significantly stronger anti-proliferative effects in HCT-116 and HT-29 cell lines (P<0.01). Compound K significantly arrested HCT-116 and HT-29 cells in the G1 phase, and induced cell apoptosis (P<0.01). By contrast, Rb1 did not markedly influence the cell cycle or apoptosis. Furthermore, compound K exerted significant anti-inflammatory effects even at low concentrations (P<0.05), while Rb1 did not have any distinct effects. The data obtained from the present study demonstrated that compound K, an intestinal microbiome metabolite of Rb1, may have a potential clinical value in the prevention of inflammatory-associated colorectal cancer.

Genome-Wide DNA Methylation Profiles of Phlegm-Dampness Constitution.

BACKGROUND/AIMS: Metabolic diseases are leading health concerns in today's global society. In traditional Chinese medicine (TCM), one body type studied is the phlegm-dampness constitution (PC), which predisposes individuals to complex metabolic disorders. Genomic studies have revealed the potential metabolic disorders and the molecular features of PC. The role of epigenetics in the regulation of PC, however, is unknown. METHODS: We analyzed a genome-wide DNA methylation in 12 volunteers using Illumina Infinium Human Methylation450 BeadChip on peripheral blood mononuclear cells (PBMCs). Eight volunteers had PC and 4 had balanced constitutions. RESULTS: Methylation data indicated a genome-scale hyper-methylation pattern in PC. We located 288 differentially methylated probes (DMPs). A total of 256 genes were mapped, and some of these were metabolic-related. SQSTM1, DLGAP2 and DAB1 indicated diabetes mellitus; HOXC4 and SMPD3, obesity; and GRWD1 and ATP10A, insulin resistance. According to Ingenuity Pathway Analysis (IPA), differentially methylated genes were abundant in multiple metabolic pathways. CONCLUSION: Our results suggest the potential risk for metabolic disorders in individuals with PC. We also explain the clinical characteristics of PC with DNA methylation features.

Expression profiling-based clustering of healthy subjects recapitulates classifications defined by clinical observation in Chinese medicine.

Differences between healthy subjects and associated disease risks are of substantial interest in clinical medicine. Based on clinical presentations, Traditional Chinese Medicine (TCM) classifies healthy people into nine constitutions: Balanced, Qi, Yang or Yin deficiency, Phlegm-dampness, Damp-heat, Blood stasis, Qi stagnation, and Inherited special constitutions. In particular, Yang and Yin deficiency constitutions exhibit cold and heat aversion, respectively. However, the intrinsic molecular characteristics of unbalanced phenotypes remain unclear. To determine whether gene expression-based clustering can recapitulate TCM-based classification, peripheral blood mononuclear cells (PBMCs) were collected from Chinese Han individuals with Yang/Yin deficiency (n = 12 each) and Balanced (n = 8) constitutions, and global gene expression profiles were determined using the Affymetrix HG-U133A Plus 2.0 array. Notably, we found that gene expression-based classifications reflected distinct TCM-based subtypes. Consistent with the clinical observation that subjects with Yang deficiency tend toward obesity, series-clustering analysis detected several key lipid metabolic genes (diacylglycerol acyltransferase (DGAT2), acyl-CoA synthetase (ACSL1), and ATP-binding cassette subfamily A member 1 (ABCA1)) to be down- and up-regulated in Yin and Yang deficiency constitutions, respectively. Our findings suggest that Yin/Yang deficiency and Balanced constitutions are unique entities in their mRNA expression profiles. Moreover, the distinct physical and clinical characteristics of each unbalanced constitution can be explained, in part, by specific gene expression signatures.

Traditional Chinese Medicine and Constitutional Medicine in China, Japan and Korea: A Comparative Study.

Traditional Chinese medicine (TCM), Japanese-Chinese medicine, and Korean Sasang constitutional medicine have common origins. However, the constitutional medicines of China, Japan, and Korea differ because of the influence of geographical culture, social environment, national practices, and other factors. This paper aimed to compare the constitutional medicines of China, Japan, and Korea in terms of theoretical origin, constitutional classification, constitution and pathogenesis, clinical applications and basic studies that were conducted. The constitutional theories of the three countries are all derived from the Canon of Internal Medicine or Treatise on Febrile and Miscellaneous Diseases of Ancient China. However, the three countries have different constitutional classifications and criteria. Medical sciences in the three countries focus on the clinical applications of constitutional theory. They all agree that different pathogenic laws that guide the treatment of diseases govern different constitutions; thus, patients with different constitutions are treated differently. The three countries also differ in terms of drug formulations and medication. Japanese medicine is prescribed only based on constitution. Korean medicine is based on treatment, in which drugs cannot be mixed. TCM synthesize the treatment model of constitution differentiation, disease differentiation and syndrome differentiation with the treatment thought of treating disease according to three categories of etiologic factors, which reflect the constitution as the characteristic of individual precision treatment. In conclusion, constitutional medicines of China, Japan, and Korea have the same theoretical origin, but differ in constitutional classification, clinical application of constitutional theory on the treatment of diseases, drug formulations and medication.

Reliability of online near-infrared spectroscopy analysis of the extraction process of Huatan Qushi decoction for phlegm-dampness constitution.

OBJECTIVE: To establish a quantitative model for the online analysis of hesperidin content in the extraction process of Huatan Qushi decoction for individuals with phlegm-dampness constitution by using near-infrared (NIR) spectroscopy and partial least squares (PLS) methods. METHODS: The reference content of hesperidin, an effective ingredient of Huatan Qushi decoction, was determined using high-performance liquid chromatography (HPLC). The modeling band was selected using synergy interval PLS (SiPLS) algorithms, and a PLS model demonstrating the relationship between NIR predictive values and HPLC reference values was established. The optimal modeling route was selected through global optimization of process trace parameters. RESULTS: The root mean square error of cross-validation was 9.8410. The coefficients of determination of the calibration set and prediction set were 0.9860 (R2cal) and 0.9458 (R2pre), respectively. The residual predictive deviation was 3.0115. The parameters demonstrated high predictive capability and reliability. The band was selected via the SiPLS method. There were seven latent variables. CONCLUSION: This study provides a favorable comprehensive evaluation method for variable selection and quality control of the Traditional Chinese Medicine extraction process.