Therapeutic Mechanism of Zhuyang Tongbian Decoction in Treating Functional Constipation: Insights from a Pilot Study Utilizing 16S rRNA Sequencing, Metagenomics, and Metabolomics

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study design

cross-sectional observational, not case-control

Citation

PMID PubMed identifier for scientific articles.

40026814

DOI Digital object identifier for electronic documents.

10.2147/IJGM.S509592

URI

Authors

Zhang X, Yin H, Yang X, Kang J, Sui N

Journal

International journal of general medicine

Year

2025

Keywords:

Zhuyang Tongbian Decoction, functional constipation, intestinal flora, intestinal inflammation, metabolite

PURPOSE: To explore the mechanism of Zhuyang Tongbian Decoction (ZTD) in treating functional constipation (FC) by observing its effects on intestinal flora composition, the metabolic function of gut microbiota, fecal short-chain fatty acid (SCFA) levels, and serum concentrations of TLR4, NF-κB, TNF-α, and IL-6 in patients with FC. PATIENTS AND METHODS: 40 patients with FC were randomly divided into the control group and the treatment group, 20 cases in each group. And 20 healthy volunteers were recruited during the same period. The control group was administered lactulose, while the treatment group was treated with ZTD. 16s RNA sequencing technology was used to compare the changes in the structure and diversity of the intestinal flora of patients before and after treatment. Changes in the levels of SCFAs in faeces and the levels of TLR4, NF-κB, TNF-α and IL-6 in serum were analysed. Metagenomics sequencing assessed microbiota metabolic functions. RESULTS: The treatment group showed a significant increase in the relative abundance of beneficial bacteria, including Bifidobacterium, Lactobacillus, and Faecalibacterium_prausnitzii (P < 0.05), whereas Desulfobacterota and Ruminococcus were significantly reduced (P < 0.05). Notably, fecal acetic and propionic acid levels were significantly higher in the treatment group (P < 0.05). Serum biomarkers TLR4, NF-κB, TNF-α, and IL-6 decreased significantly (P < 0.05). Metagenomics sequencing showed that Carbohydrate metabolism, Metabolism of cofactors and vitamins, and C5- Branched dibasic acid metabolism were significantly increased in functional abundance (P < 0.05). CONCLUSION: ZTD notably improves intestinal flora composition and gut microbiota metabolic function, regulates SCFA levels, and reduces inflammation markers in FC patients. The strain Faecalibacterium_prausnitzii shows significant potential in regulation of intestinal inflammation and may play a crucial role in the treatment efficacy of ZTD for FC.

Experiment 1

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Curated date: 2025/03/21

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke

Subjects

Location of subjects

China

Host species Species from which microbiome was sampled. Contact us to have more species added.

Homo sapiens

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology

Feces Cow dung,Cow pat,Droppings,Dung,Excrement,Excreta,Faeces,Fecal material,Fecal matter,Fewmet,Frass,Guano,Matières fécales@fr,Merde@fr,Ordure,Partie de la merde@fr,Piece of shit,Porción de mierda@es,Portion of dung,Portion of excrement,Portion of faeces,Portion of fecal material,Portion of fecal matter,Portion of feces,Portion of guano,Portion of scat,Portionem cacas,Scat,Spoor,Spraint,Stool,Teil der fäkalien@de,Feces,feces

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology

Treatment outcome measurement Treatment outcome measurement,treatment outcome measurement

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy Volunteers)

Group 1 name Corresponds to the case (exposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group was treated with Zhuyang Tongbian Decoction (ZTD) (raw drug concentration: 3.0 g/mL)

Group 0 sample size Number of subjects in the control (unexposed) group

20

Group 1 sample size Number of subjects in the case (exposed) group

20

Antibiotics exclusion Number of days without antibiotics usage (if applicable) and other antibiotics-related criteria used to exclude participants (if any)

1 month

Lab analysis

Sequencing type

16S

16S variable region One or more hypervariable region(s) of the bacterial 16S gene

V3-V4

Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance

Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).

relative abundances

Significance threshold p-value or FDR threshold used for differential abundance testing (if any)

0.05

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?

No

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

decreased

Signature 1

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Curated date: 2025/03/21

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke

Source: Fig 2c

Description: Comparison of significantly different flora before and after treatment with ZTD at the genus level

Abundance in Group 1: increased abundance in FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

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Collinsella
Desulfovibrio
Romboutsia
Ruminococcus
UCG-002UCG-002
rumen bacterium NK4A214

Revision editor(s): PreciousChijioke

Signature 2

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Source: Fig 2c

Description: Comparison of significantly different flora before and after treatment with ZTD at the genus level

Abundance in Group 1: decreased abundance in FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

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Agathobacter
Faecalibacterium
Fusicatenibacter
Subdoligranulum
Christensenellaceae_R_7_groupChristensenellaceae_R_7_group

Revision editor(s): PreciousChijioke

Experiment 2

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

Group 1 name Corresponds to the case (exposed) group for case-control studies

ProZY (Pro-Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group after treatment with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

increased

Signature 1

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Source: Fig 2c

Description: Comparison of significantly different flora before and after treatment with ZTD at the genus level

Abundance in Group 1: increased abundance in ProZY (Pro-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
Agathobacter
Faecalibacterium
Fusicatenibacter
Subdoligranulum
Christensenellaceae_R_7_groupChristensenellaceae_R_7_group

Revision editor(s): PreciousChijioke

Signature 2

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Source: Fig 2c

Description: Comparison of significantly different flora before and after treatment with ZTD at the genus level

Abundance in Group 1: decreased abundance in ProZY (Pro-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
Collinsella
Desulfovibrio
Romboutsia
Ruminococcus
UCG-002UCG-002
rumen bacterium NK4A214

Revision editor(s): PreciousChijioke

Experiment 3

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Curated date: 2025/03/22

Curator: PreciousChijioke

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) + ProZY (Pro-Zhuyang Tongbian Decoction)

Group 1 name Corresponds to the case (exposed) group for case-control studies

NC (Healthy volunteers)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Healthy volunteers with Bristol faecal character score of 4 according to the scoring criteria

Lab analysis

Statistical Analysis

Statistical test

LEfSe

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

2

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Source: Fig 4

Description: Analysis of differential flora at different taxonomic levels

Abundance in Group 1: increased abundance in NC (Healthy volunteers)

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Adlercreutzia
Agathobacter
Allisonella
Bacillota
Dialister
Dorea
Faecalibacterium
Faecalibacterium prausnitzii
Fusicatenibacter
Lachnoclostridium
Lachnospiraceae
Lachnospirales
Lactobacillaceae
Lactobacillus
Megamonas funiformis
Negativicutes
Oscillospiraceae
Roseburia
Ruminococcus gauvreauii
Veillonellaceae
gut metagenome
Veillonellales-SelenomonadalesVeillonellales-Selenomonadales

Revision editor(s): PreciousChijioke, KateRasheed

Experiment 4

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Curated date: 2025/03/22

Curator: PreciousChijioke

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy volunteers) + ProZY (Pro-Zhuyang Tongbian Decoction)

Group 1 name Corresponds to the case (exposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group was treated with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Signature 1

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Source: Fig 4

Description: Analysis of differential flora at different taxonomic levels

Abundance in Group 1: increased abundance in FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction)

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Clostridia
Clostridiaceae
Colidextribacter
Collinsella
Coriobacteriaceae
Coriobacteriales
Coriobacteriia
Desulfovibrio
Desulfovibrionaceae
Desulfovibrionales
Desulfovibrionia
Eggerthellaceae
Erysipelotrichaceae
Erysipelotrichales
Holdemanella
Intestinibacter
Muribaculaceae
Oscillospiraceae
Peptostreptococcaceae
Romboutsia
Ruminococcus callidus
Senegalimassilia
Streptococcus equinus
Terrisporobacter
rumen bacterium NK4A214
UCG_002UCG_002
Eubacteriales
Eubacterium coprostanoligenes
Peptostreptococcales-TissierellalesPeptostreptococcales-Tissierellales
Thermodesulfobacteriota
Clostridium
UCG_005UCG_005
[Ruminococcus] torques

Revision editor(s): PreciousChijioke, KateRasheed

Experiment 5

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Curated date: 2025/03/22

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke, KateRasheed

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy volunteers) + FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction)

Group 1 name Corresponds to the case (exposed) group for case-control studies

ProZY (Pro-Zhuyang Tongbian Decoction)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group after treatment with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Signature 1

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Curated date: 2025/03/23

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Source: Fig 4

Description: Analysis of differential flora at different taxonomic levels

Abundance in Group 1: increased abundance in ProZY (Pro-Zhuyang Tongbian Decoction)

NCBI	Quality Control	Links
Bacteroides clarus
Butyricicoccaceae
Butyricicoccus
Christensenellaceae
Christensenellales
Lachnospira
Lactobacillales
Marinifilaceae
Oscillibacter
Paraprevotella
Streptococcaceae
Streptococcus
Streptococcus salivarius
Veillonella
Veillonella parvula
gut metagenome
Christensenellaceae_R_7_groupChristensenellaceae_R_7_group
UCG_008UCG_008
Clostridia UCG_014Clostridia UCG_014
Bacilli
Lachnospiraceae bacterium NK4A136
Lachnospira eligens
Prevotellaceae NK3B31 groupPrevotellaceae NK3B31 group
Enterococcaceae bacterium RF39
Lachnospiraceae UCG-001Lachnospiraceae UCG-001
[Eubacterium] siraeum

Revision editor(s): PreciousChijioke, KateRasheed

Experiment 6

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Curated date: 2025/03/27

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke, KateRasheed

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy Volunteers)

Group 1 name Corresponds to the case (exposed) group for case-control studies

FC_PreZY (Functional Constipation in Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group was treated with Zhuyang Tongbian Decoction (ZTD) (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Statistical test

Mann-Whitney (Wilcoxon)

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

Not specified

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

decreased

Signature 1

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Curated date: 2025/03/28

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: decreased abundance in FC_PreZY (Functional Constipation in Zhuyang Tongbian Decoction) group

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Bifidobacterium
Lactobacillus

Revision editor(s): PreciousChijioke

Experiment 7

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

FC_Prezy (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

Group 1 name Corresponds to the case (exposed) group for case-control studies

ProZY (Pro-Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group after treatment with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

increased

Signature 1

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: increased abundance in ProZY (Pro-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
Bifidobacterium
Lactobacillus

Revision editor(s): PreciousChijioke

Experiment 8

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Curator: PreciousChijioke

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy Volunteers)

Group 1 name Corresponds to the case (exposed) group for case-control studies

FC_PreCG (Functional Constipation in Pre-Control Group)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreCG group was treated with lactulose (manufacturer: Fresenius Kabi Austria GmbH, Austria) orally

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

unchanged

Signature 1

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Curated date: 2025/03/29

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: decreased abundance in FC_PreCG (Functional Constipation in Pre-Control Group)

NCBI	Quality Control	Links
Bifidobacterium
Lactobacillus

Revision editor(s): PreciousChijioke

Experiment 9

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

FC_PreCG (Functional Constipation in Pre-Control Group)

Group 1 name Corresponds to the case (exposed) group for case-control studies

ProCG (Post-Control Group)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreCG group after treatment with lactulose orally

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

increased

Signature 1

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: increased abundance in ProCG (Post-Control Group)

NCBI	Quality Control	Links
Bifidobacterium

Revision editor(s): PreciousChijioke

Signature 2

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: decreased abundance in ProCG (Post-Control Group)

NCBI	Quality Control	Links
Lactobacillus

Revision editor(s): PreciousChijioke

Experiment 10

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

ProZY (Pro-Zhuyang Tongbian Decoction)

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

unchanged

Signature 1

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Curated date: 2025/04/04

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Source: Fig 3a

Description: Comparison of the abundance of Bifidobacterium spp. and Lactobacillus spp. in each group

Abundance in Group 1: increased abundance in ProCG (Post-Control Group)

NCBI	Quality Control	Links
Bifidobacterium

Revision editor(s): PreciousChijioke

Experiment 11

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Curated date: 2025/04/04

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke, KateRasheed

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

NC (Healthy Volunteers)

Group 1 name Corresponds to the case (exposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group was treated with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

increased

Signature 1

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Curated date: 2025/04/04

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Revision editor(s): PreciousChijioke

Source: Fig 3c

Description: Comparison of significantly different flora before and after treatment with ZTD at the species level

Abundance in Group 1: increased abundance in FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
uncultured bacterium

Revision editor(s): PreciousChijioke

Signature 2

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Curated date: 2025/04/05

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Source: Fig 3c

Description: Comparison of significantly different flora before and after treatment with ZTD at the species level

Abundance in Group 1: decreased abundance in FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
Escherichia coli
Bifidobacterium adolescentis
Faecalibacterium prausnitzii
Bifidobacterium longum
Bifidobacterium pseudocatenulatum
gut metagenome

Revision editor(s): PreciousChijioke

Experiment 12

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

FC_PreZY (Functional Constipation in Pre-Zhuyang Tongbian Decoction) group

Group 1 name Corresponds to the case (exposed) group for case-control studies

ProZY (Pro-Zhuyang Tongbian Decoction) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

FC_PreZY group after treatment with ZTD (raw drug concentration: 3.0 g/mL)

Lab analysis

Statistical Analysis

Alpha Diversity

Faith Phylogenetic diversity, takes into account phylogenetic distance of all taxa identified in a sample

increased

Signature 1

EditHistoryDelete

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Reviewed Marked as Reviewed by Svetlana up on 2025-4-7

Curated date: 2025/04/04

Curator: PreciousChijioke

Revision editor(s): PreciousChijioke, KateRasheed

Source: Fig 3c

Description: Comparison of significantly different flora before and after treatment with ZTD at the species level

Abundance in Group 1: decreased abundance in ProZY (Pro-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
uncultured bacterium

Revision editor(s): PreciousChijioke, KateRasheed

Signature 2

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Curated date: 2025/04/05

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Revision editor(s): PreciousChijioke, KateRasheed

Source: Fig 3c

Description: Comparison of significantly different flora before and after treatment with ZTD at the species level

Abundance in Group 1: increased abundance in ProZY (Pro-Zhuyang Tongbian Decoction) group

NCBI	Quality Control	Links
Bifidobacterium adolescentis
Bifidobacterium longum
Bifidobacterium pseudocatenulatum
Escherichia coli
Faecalibacterium prausnitzii
gut metagenome

Revision editor(s): PreciousChijioke, KateRasheed