Fig. 1.
Overview of the retrospective cohort study design.
This flow chart presents the study's framework, focusing on a retrospective cohort of hospitalized ulcerative colitis patients, categorized upon admission and subsequent fecal microbiota transplantation (FMT) treatment. The study also includes placebo groups partitioned by gender. Four categories were devised: male and female patients receiving FMT and male and female patients receiving a placebo.
Table I
Baseline characteristics of ulcerative colitis (UC) patients.
| Parameters | Experimental group (n = 68) | Placebo group (n = 68) | χ2 or t-values | p-values |
|---|---|---|---|---|
| Males | ||||
| Age | 45.2 ± 13.6 | 47.1 ± 11.4 | 0.402 | 0.688 |
| BMI | 22.5 ± 2.16 | 21.5 ± 3.4 | 1.484 | 0.141 |
| Smoker, n (%) | 18 (43.9%) | 17 (25.0%) | 0.038 | 0.844 |
| Drinker, n (%) | 12 (17.6%) | 11 (16.2%) | 0.052 | 0.819 |
| Protein intake (g/day) | 79.252 ± 14.791 | 82.543 ± 27.594 | 0.456 | 0.377 |
| Fat intake (g/day) | 89.618 ± 33.173 | 92.609 ± 38.641 | 0.158 | 0.623 |
| Carbohydrate intake (g/day) | 258.167 ± 70.847 | 264.875 ± 74.136 | 0.568 | 0.326 |
| Vitamin A intake (μg/day) | 817.949 ± 190.225 | 839.24 ± 196.633 | 0.479 | 0.354 |
| Vitamin C intake (mg/day) | 99.564 ± 37.648 | 108.344 ± 11.862 | 0.984 | 0.102 |
| Vitamin D intake (μg/day) | 28.134 ± 14.025 | 30.848 ± 10.064 | 1.033 | 0.096 |
| Calcium intake (mg/day) | 997.56 ± 202.16 | 1018.256 ± 310.141 | 1.527 | 0.058 |
| Iron intake (mg/day) | 41.123 ± 25.249 | 38.897 ± 35.102 | 1.320 | 0.079 |
| hsCRP (mg/dl) | 3.707 ± 1.209 | 4.034 ± 1.25 | 0.135 | 0.893 |
| Triglyceride (mg/dl) | 98.2 ± 33.0 | 95.4 ± 33.9 | 1.375 | 0.172 |
| Total cholesterol (mg/dl) | 188 ± 21.4 | 191.6 ± 14.3 | 0.841 | 0.402 |
| HDL-c (mg/dl) | 49.7 ± 6.11 | 46.04 ± 7.44 | 1.951 | 0.064 |
| LDL-c (mg/dl) | 123 ± 18 | 129.9 ± 21.3 | 1.152 | 0.133 |
| Females | ||||
| Age | 40.6 ± 12.9 | 36.7 ± 14.0 | 1.220 | 0.225 |
| BMI | 21.6 ± 1.94 | 20.55 ± 1.62 | 1.861 | 0.056 |
| Smoker, n (%) | 20 (48.78%) | 18 (26.5%) | 0.146 | 0.702 |
| Drinker, n (%) | 11 (16.17%) | 13 (19.12%) | 0.202 | 0.653 |
| Menopausal status | ||||
| Premenopausal, n (%) | 41 (60.29) | 48 (70.59) | ||
| Perimenopausal, n (%) | 20 (29.41) | 17 (25) | 2.394 | 0.302 |
| Postmenopausal, n (%) | 7 (10.29) | 3 (4.41) | ||
| Protein intake (g/day) | 75.252 ± 28.791 | 70.543 ± 15.594 | 1.632 | 0.053 |
| Fat intake (g/day) | 73.618 ± 32.173 | 77.609 ± 16.641 | 0.105 | 0.647 |
| Carbohydrate intake (g/day) | 247.167 ± 74.847 | 259.875 ± 85.136 | 0.258 | 0.394 |
| Vitamin A intake (μg/day) | 830.949 ± 184.225 | 870.24 ± 220.633 | 0.752 | 0.214 |
| Vitamin C intake (mg/day) | 115.564 ± 35.648 | 112.344 ± 32.862 | 0.121 | 0.698 |
| Vitamin D intake (μg/day) | 12.134 ± 4.975 | 10.848 ± 5.064 | 0.097 | 0.876 |
| Calcium intake (mg/day) | 1000.56 ± 222.16 | 1096.256 ± 307.141 | 0.118 | 0.7063 |
| Iron intake (mg/day) | 12.123 ± 10.249 | 13.897 ± 6.102 | 0.237 | 0.425 |
| hsCRP (mg/dl) | 3.776 ± 1.379 | 3.694 ± 1.663 | 0.023 | 0.982 |
| Triglyceride (mg/dl) | 99.9 ± 31.2 | 99.4 ± 33.8 | 0.453 | 0.651 |
| Total cholesterol (mg/dl) | 193 ± 19.3 | 186.4 ± 26.8 | 0.781 | 0.436 |
| HDL-c (mg/dl) | 50.7 ± 5 | 49.7 ± 1.5 | 0.504 | 0.616 |
| LDL-c (mg/dl) | 119 ± 18.1 | 119.5 ± 19.8 | 0.753 | 0.453 |
[i] The statistical difference is insignificant if p > 0.05.
Table II
Baseline characteristics of adolescent donors.
| Parameters | Males (n = 68) | Females (n = 68) | t-values | p-values |
|---|---|---|---|---|
| Age | 13.91 ± 2.22 | 13.91 ± 2.563 | 0.010 | 0.992 |
| BMI | 22.59 ± 1.468 | 22.38 ± 1.416 | 0.001 | 0.999 |
| hsCRP (mg/dl) | 0.487 ± 0.253 | 0.4856 ± 0.299 | 1.232 | 0.999 |
| Triglyceride (mg/dl) | 73.24 ± 14.41 | 75.41 ± 14.96 | 0.821 | 0.771 |
| Total cholesterol (mg/dl) | 169.6 ± 14.09 | 171.1 ± 13.43 | 0.268 | 0.958 |
| HDL-c (mg/dl) | 64.91 ± 3.155 | 65.38 ± 2.962 | 0.513 | 0.999 |
| LDL-c (mg/dl) | 94.8 ± 15.2 | 95.7 ± 13.67 | 0.002 | 0.996 |
[i] The statistical difference is insignificant if p > 0.05.
Table III
UC scores before and after FMT (± SD).
| Time | Diarrhea score | Abdominal pain score | Bloody stool | Mucosa score | Mayo score |
|---|---|---|---|---|---|
| Males (experimental group) | |||||
| Before FMT | 5.294 ± 1.658 | 5.091 ± 1.696 | 5.358 ± 1.579 | 1.772 ± 0.516 | 5.357 ± 1.724 |
| 3 weeks | 4.719 ± 1.612 *, ## | 4.598 ± 1.378 *, ## | 3.956 ± 1.428 **, ***, #### | 1.533 ± 0.478 *, # | 5.066 ± 1.651 |
| 6 weeks | 4.267 ± 1.457 ***, #, ## | 4.269 ± 1.428 ***, #, ## | 3.932 ± 1.195 **, ***, #### | 1.416 ± 0.441 ***, #, ## | 4.317 ± 1.806 ***, #, ## |
| 9 weeks | 3.965 ± 1.163 ****, #### | 4.116 ± 1.053 ****, #### | 3.524 ± 0.895 **, ***, #### | 1.259 ± 0.385 ****, #### | 4.443 ± 1.242 **, ## |
| Males (placebo group) | |||||
| Before FMT | 5.511 ± 1.605 | 5.089 ± 2.126 | 5.464 ± 1.217 | 1.837 ± 0.772 | 5.360 ± 1.565 |
| 3 weeks | 5.430 ± 1.937 | 5.246 ± 1.725 | 5.815 ± 1.391 | 1.960 ± 0.681 | 5.405 ± 2.220 |
| 6 weeks | 5.504 ± 1.351 | 5.305 ± 1.892 | 5.194 ± 1.432 | 1.859 ± 0.546 | 5.483 ± 2.218 |
| 9 weeks | 5.566 ± 2.071 | 5.188 ± 1.762 | 5.073 ± 1.229 | 1.772 ± 0.508 | 4.957 ± 1.361 |
| Females (experimental group) | |||||
| Before FMT | 4.979 ± 1.741 | 4.812 ± 1.606 | 5.086 ± 1.696 | 1.721 ± 0.501 | 5.665 ± 1.808 |
| 3 weeks | 4.005 ± 1.282 ****, #, ## | 3.381 ± 1.327 ****, #### | 3.520 ± 1.254 ****, #### | 1.442 ± 0.457 **, ## | 4.730 ± 1.473 ***, #, ## |
| 6 weeks | 3.523 ± 1.087 ****, #, ## | 3.12 ± 1.009 ****, #### | 3.034 ± 0.925 ****, #### | 1.24 ± 0.349 ****, #### | 4.503 ± 1.643 ****, #### |
| 9 weeks | 2.933 ± 0.787 ****, #### | 2.924 ± 0.835 ****, #### | 2.396 ± 0.535 ****, #### | 1.098 ± 0.309 ****, #### | 3.978 ± 1.096 ****, #### |
| Females (placebo group) | |||||
| Before FMT | 5.318 ± 2.286 | 5.060 ± 1.895 | 5.009 ± 1.574 | 1.653 ± 0.752 | 5.711 ± 1.858 |
| 3 weeks | 5.095 ± 1.952 | 4.512 ± 1.217 | 5.587 ± 2.193 | 1.958 ± 0.363 | 5.287 ± 2.241 |
| 6 weeks | 4.640 ± 1.970 | 5.282 ± 1.496 | 5.079 ± 1.563 | 1.867 ± 0.743 | 5.533 ± 2.010 |
| 9 weeks | 5.459 ± 1.997 | 5.062 ± 1.390 | 5.048 ± 2.109 | 1.713 ± 0.654 | 5.542 ± 1.831 |
Table IV
Self-rating depression scale (SDS) and self-rating anxiety scale (SAS) before and after FMT (± SD).
| Time | Experimental group | Placebo group | ||
|---|---|---|---|---|
| SDS | SAS | SDS | SAS | |
| Males | ||||
| Before FMT | 55.29 ± 7.393 | 44.16 ± 5.929 | 57.829 ± 9.209 | 43.377 ± 4.004 |
| 3 weeks | 49.55 ± 6.093 ***, #, ## | 41.01 ± 5.287 # | 55.113 ± 5.801 | 41.958 ± 7.969 |
| 6 weeks | 45.98 ± 5.4 ****, #### | 37.5 ± 4.275 ***, #, ## | 58.437 ± 8.857 | 42.09 ± 7.982 |
| 9 weeks | 42.84 ± 4.051 ****, #### | 33.99 ± 3.623 ****, #### | 55.386 ± 11.322 | 45.402 ± 6.562 |
| Females | ||||
| Before FMT | 54.76 ± 7.411 | 45.09 ± 5.935 | 55.217 ± 9.891 | 44.784 ± 5.301 |
| 3 weeks | 46.79 ± 5.752 ****, #, ## | 39.08 ± 5.17 ***, ## | 54.6 ± 10.427 | 44.177 ± 8.28 |
| 6 weeks | 43.41 ± 5.362 ****, #, ## | 34.29 ± 3.909 ****, ## | 53.23 ± 6.527 | 46.833 ± 7.012 |
| 9 weeks | 37.66 ± 4.091 ****, #### | 28.5 ± 3.781 ****, #### | 56.04 ± 5.384 | 49.115 ± 4.044 |
Table V
hsCRP levels in ulcerative colitis (UC) patients before and after FMT (± SD).
| Time | Experimental group | Placebo group | t-values | p-values |
|---|---|---|---|---|
| Males | ||||
| Before FMT | 3.707 ± 1.209 | 3.634 ± 1.262 | 0.288 | 0.294 |
| 3 weeks | 3.521 ± 1.312 | 4.127 ± 1.374 | 3.279 | 0.001 |
| 6 weeks | 3.511 ± 1.091 | 3.493 ± 0.966 | 0.134 | 0.894 |
| 9 weeks | 3.033 ± 1.184** | 3.577 ± 1.581 | 1.753 | 0.008 |
| Females | ||||
| Before FMT | 3.776 ± 1.379 | 3.426 ± 1.295 | 1.941 | 0.054 |
| 3 weeks | 3.219 ± 1.219* | 3.791 ± 1.275 | 2.035 | 0.044 |
| 6 weeks | 2.821 ± 1.023**** | 3.474 ± 1.091 | 3.911 | 0.000 |
| 9 weeks | 2.35 ± 0.8592**** | 3.526 ± 1.365 | 6.359 | 0.000 |
Fig. 2 A, B.
Comparative analysis of microbiota characteristics among patient cohorts.
A – via a Venn diagram, represents the unique and shared operational taxonomic units (OTUs) for male patients in both experimental and placebo groups. B – similarly represents female patients. Overlapping circles demonstrate shared OTUs.
AFP represents female ulcerative colitis patients post-placebo treatment; AM signifies male ulcerative colitis patients post-fecal microbiota transplantation (FMT); AMP indicates male patients post-placebo treatment. AS refers to a subgroup of 20 male ulcerative colitis patients post-FMT, while AU represents a similar subgroup of 20 female patients post-FMT. BF shows data from female ulcerative colitis patients’ pre-FMT, and BFP is for those same female patients pre-placebo treatment. BM and BMP correspond to male ulcerative colitis patients’ pre-FMT and pre-placebo treatment. BS and BU denote subgroups of 20 male and female patients pre-FMT. Finally, FD and MD represent female and male fecal donors, respectively.
Fig. 2 C, D, E, F, G.
Comparative analysis of microbiota characteristics among patient cohorts.
C and D – exhibit the rank abundance curves based on relative species abundance and biodiversity components for male and female patients, respectively. E, F, and G – demonstrate the top 10 bacterial families for male and female donors and male and female patients in both experimental and placebo groups, respectively.
AFP represents female ulcerative colitis patients post-placebo treatment; AM signifies male ulcerative colitis patients post-fecal microbiota transplantation (FMT); AMP indicates male patients post-placebo treatment. AS refers to a subgroup of 20 male ulcerative colitis patients post-FMT, while AU represents a similar subgroup of 20 female patients post-FMT. BF shows data from female ulcerative colitis patients’ pre-FMT, and BFP is for those same female patients pre-placebo treatment. BM and BMP correspond to male ulcerative colitis patients’ pre-FMT and pre-placebo treatment. BS and BU denote subgroups of 20 male and female patients pre-FMT. Finally, FD and MD represent female and male fecal donors, respectively.
Fig. 3A, B.
Alpha and beta diversity analysis of gut microbiota among patient cohorts.
This figure depicts various indices measuring microbial diversity within and between cohorts. A–D represent diversity indices (Shannon, Simpson, ACE, Chao1) for male patients across experimental and placebo groups. E–H mirror these indices for female patients. I–L illustrate Principal Component Analysis (PCA) and Principal Coordinate Analysis (PCoA) for male and female patients, respectively, comparing overall gut microbiota structure with FMT or placebo treatments. Ellipses represent an 80% confidence level, with color/shape indicating the specific cohort. AM and AF denote male and female ulcerative colitis patients, post-FMT; BF and BM are used to indicate female and male ulcerative colitis patients, respectively, pre-FMT. AFP and AMP represent female and male patients, respectively, post-placebo treatment, while BFP and BMP signify female and male patients, respectively, pre-placebo treatment.
Fig. 3C, D, E, F, G, H.
Alpha and beta diversity analysis of gut microbiota among patient cohorts.
Fig. 3I, J, K, L.
Alpha and beta diversity analysis of gut microbiota among patient cohorts.
This figure depicts various indices measuring microbial diversity within and between cohorts. A–D represent diversity indices (Shannon, Simpson, ACE, Chao1) for male patients across experimental and placebo groups. E–H mirror these indices for female patients. I–L illustrate Principal Component Analysis (PCA) and Principal Coordinate Analysis (PCoA) for male and female patients, respectively, comparing overall gut microbiota structure with FMT or placebo treatments. Ellipses represent an 80% confidence level, with color/shape indicating the specific cohort. AM and AF denote male and female ulcerative colitis patients, post-FMT; BF and BM are used to indicate female and male ulcerative colitis patients, respectively, pre-FMT. AFP and AMP represent female and male patients, respectively, post-placebo treatment, while BFP and BMP signify female and male patients, respectively, pre-placebo treatment.
Fig. 4A.
Heatmaps illustrating microbiome analysis results.
This figure showcases heatmaps of the top-ranking gut microbiota OTUs among patients before and after FMT or placebo treatments.
A – the top 35 OTUs in male patients before and after FMT. AM and AF denote male and female ulcerative colitis patients, respectively, post-FMT; AS refers to a subgroup of 20 male ulcerative colitis patients post-FMT, while AU represents a similar subgroup of 20 female patients post-FMT. BU and BS are used to indicate female and male ulcerative colitis patients, respectively, pre-FMT. AFP and AMP represent female and male patients, respectively, post-placebo treatment, while BFP and BMP signify female and male patients, respectively, pre-placebo treatment.
Fig. 4B, C.
Heatmaps illustrating microbiome analysis results.
This figure showcases heatmaps of the top-ranking gut microbiota OTUs among patients before and after FMT or placebo treatments. B – the male group pre- and post-placebo. C – the female patients with the top 45 OTUs before and after FMT.
AM and AF denote male and female ulcerative colitis patients, respectively, post-FMT; AS refers to a subgroup of 20 male ulcerative colitis patients post-FMT, while AU represents a similar subgroup of 20 female patients post-FMT. BU and BS are used to indicate female and male ulcerative colitis patients, respectively, pre-FMT. AFP and AMP represent female and male patients, respectively, post-placebo treatment, while BFP and BMP signify female and male patients, respectively, pre-placebo treatment.
Fig. 4C.
Heatmaps illustrating microbiome analysis results.
C – the female patients with the top 45 OTUs before and after FMT.
Fig. 4D.
Heatmaps illustrating microbiome analysis results.
This figure showcases heatmaps of the top-ranking gut microbiota OTUs among patients before and after FMT or placebo treatments. D – the female group pre- and post-placebo.
AM and AF denote male and female ulcerative colitis patients, respectively, post-FMT; AS refers to a subgroup of 20 male ulcerative colitis patients post-FMT, while AU represents a similar subgroup of 20 female patients post-FMT. BU and BS are used to indicate female and male ulcerative colitis patients, respectively, pre-FMT. AFP and AMP represent female and male patients, respectively, post-placebo treatment, while BFP and BMP signify female and male patients, respectively, pre-placebo treatment.