Application of phytobacterium plantarum chen-02 in preparation of medicine for improving intestinal dysfunction caused by burn

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of plant lactobacillus chen-02 in preparation of a medicine for improving intestinal dysfunction caused by burn. The lactobacillus plantarum chen-02 is preserved in China General Microbiological Culture Collection Center (CGMCC) on September 13, 2021, the address is No.3, No.1 yard, Beichen West Road, Chaoyang District, Beijing, and the preservation number is CGMCC NO.23397. The lactobacillus plantarum chen-02 is named as Lactobacillus plantarum chen-02. The lactobacillus plantarum chen-02 is named as Lactobacillus plantarum chen-02. The lactobacillus plantarum chen-02 is named as Lactobacillus plantarum chen-02. The plant lactobacillus chen-02 provided by the invention can promote the balance recovery of intestinal flora of a serious burn patient, change the composition of the intestinal flora, increase the abundance of probiotics and reduce the abundance of opportunistic pathogenic bacteria, so that the intestinal function of the serious burn patient is improved, and the wound healing of the serious burn patient is further promoted.

Description

Technical Field

[0001] The present application belongs to the field of biomedicine technology, and specifically relates to the use of a strain of Lactobacillus plantarum chen-02 in the preparation of a drug for improving intestinal dysfunction caused by burns. Background Art

[0002] Burns are injuries to the skin or mucous membranes caused by factors such as high temperature, electric current, corrosive substances, or radiation. The severity of a burn depends on the depth and size of the wound, which is generally categorized into four degrees: first-degree, superficial second-degree, deep second-degree, and third-degree. Severe burns can cause direct damage to the burn site in the early stages, followed by a series of systemic cascade reactions. Pathological and physiological changes in various organs and systems can lead to complications, even catastrophic consequences such as sepsis, shock, and multiple organ failure. Intestinal dysfunction is a common complication of burns, primarily due to intestinal dysbiosis and disruption of the intestinal mucosal barrier.

[0003] After severe burns, the intestinal microbiome is severely imbalanced. Firmicutes decrease in abundance and Bacteroidetes increase in abundance. In the early stages of burns, the proportion of Gram-negative bacteria increases, particularly opportunistic pathogens such as Escherichia coli and Shigella. Meanwhile, certain probiotics, particularly those producing short-chain fatty acids (SCFAs) and lactic acid, such as Clostridium butyricum, Clostridium difficile, and Bifidobacterium, decrease to varying degrees. Intestinal dysbiosis can disrupt the intestinal environment, further triggering intestinal inflammation and immune dysfunction, impairing intestinal function and even leading to bacterial translocation (BT) and endotoxin translocation, leading to serious complications such as infection, sepsis, and multiple organ dysfunction syndrome (MODS). These complications can affect the treatment outcome, length of hospital stay, and even the lifespan of patients with severe burns. Therefore, restoring intestinal microbial balance is crucial for improving the prognosis of patients with severe burns. Summary of the Invention

[0004] The object of the present invention is to overcome the deficiencies in the prior art and provide a plant lactobacillus and its application, specifically adopting the following technical solutions: In the first aspect, a strain of Lactobacillus plantarum chen-02 is used in the preparation of a drug for improving intestinal dysfunction caused by burns, wherein the Lactobacillus plantarum ( Lactobacillus plantarum )chen-02 was deposited on September 13, 2021 at the General Microbiology Center of China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC NO. 23397.

[0005] In a second aspect, the present invention provides a biological preparation, the active ingredient of which includes the above-mentioned Lactobacillus plantarum chen-02.

[0006] In a third aspect, the present invention provides a drug for improving intestinal dysfunction caused by burns, wherein the drug comprises the aforementioned Lactobacillus plantarum chen-02 or the aforementioned biological preparation.

[0007] As a further preferred embodiment, the drug further comprises pharmaceutical excipients.

[0008] As a further preferred embodiment, the pharmaceutical excipient is at least one of water, lactose, sodium chloride, and glucose.

[0009] As a further preferred embodiment, the dosage form of the drug is powder, granules, capsules or tablets.

[0010] As a further preferred embodiment, the drug is an oral drug.

[0011] In a fourth aspect, the present invention provides the use of the aforementioned Lactobacillus plantarum in the preparation of a product for improving and / or treating burn wound healing.

[0012] As a further preferred embodiment, the product includes a drug or a biological preparation.

[0013] In the fifth aspect, the present invention provides a drug for improving and / or treating burn wound healing, wherein the drug includes Lactobacillus plantarum chen-02, which has been deposited in the General Microbiology Center of China Culture Collection Administration on September 13, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with a deposit number of CGMCC NO.23397.

[0014] The beneficial effects of the present invention are: The present invention provides a strain of Lactobacillus plantarum, which can promote the restoration of intestinal flora balance in patients with severe burns, change the composition of intestinal flora, increase the abundance of probiotics and reduce the abundance of opportunistic pathogens, thereby improving the intestinal function of patients with severe burns and further promoting wound healing in patients with severe burns. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1Shown is the comparison of intestinal dysfunction symptom scores between the two groups at different time points; Figure 2 Shown are gel images of amplification detection of some samples; Figure 3 Shown is the Venn diagram of ASV at different time points in two groups; Figure 4 Shown is the dilution curve for a stool sample; Figure 5 Shown is the abundance rank curve for fecal samples; Figure 6 Shown is the comparison of intestinal flora α diversity between the two groups of patients at different time points; Figure 7 Shown is the comparison of intestinal flora β diversity between the two groups of patients at different time points; Figure 8 Shown is the analysis of differences between groups based on the anosim matrix test method; Figure 9 Shown is a composition diagram at the species phylum level; Figure 10 Shown is a comparison of relative abundance at the phylum level; Figure 11 Shown is a comparison of relative abundance at the species genus level; Figure 12 Shown is a comparison of relative abundances of major species at the genus level; Figure 13 Shown is a species clustering heat map; Figure 14 Shown is a tree diagram of the species classification hierarchy; Figure 15 Shown is the comparison of APACHE II scores between the two groups of patients at different times [M(P25, P75)]; Figure 16 Shown is the comparison of wound healing rates between the two groups on day 14; Note: In the above figures, TPD0: before intervention of TP group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TP group; TSD0: before intervention of TS group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TS group. DETAILED DESCRIPTION

[0017] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are part of the embodiments of this application, not all of them. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0018] The plant lactobacillus used in the present invention Lactobacillus plantarum )chen-02 has been declared in the existing technology with patent number 202310707272.4 and is an existing strain.

[0019] Example 1 Lactobacillus plantarum ( Lactobacillus plantarum ) Evaluation of the effect of chen-02 on intestinal dysfunction after burns 1. Research subjects: This invention selected patients with severe burns who were hospitalized in the burn department of a certain hospital from May 2023 to March 2025 as research subjects. The subjects understood and signed the relevant treatment consent form.

[0020] 2. Sample and data sources: The general clinical data in this article were all obtained from the electronic medical record system of a certain hospital, and 16S rRNA sequencing was completed by Shanghai Paisonno Biotechnology Co., Ltd.

[0021] 3. Ethical Statement: This invention has been approved by the ethics committee of a certain hospital [Ethics Number: IIT

[2023] Lin Lun Shen No. 168]. The subjects included in the research were aware of the content of this study and have signed the informed consent.

[0022] 4. Inclusion criteria: (1) aged 18-65 years, with good general health history; (2) admitted to a hospital burn department within 72 hours after burn and hospitalized; (3) severely burned patients (total burn area >30% TBSA (total body surface area) or third-degree burn area >10% TBSA); (4) agreed to participate and signed the informed consent form.

[0023] 5. Intervention measures (1) Conventional treatment: After admission, the patient's vital signs are monitored, relevant tests and examinations are completed, and timely fluid replacement is performed to prevent shock, and water, electrolyte and acid-base imbalances are corrected. Broad-spectrum antibiotics and antifungal drugs are used to prevent infection, and proton pump inhibitors are used to prevent stress ulcers. According to the patient's specific condition, endotracheal intubation, tracheotomy, mechanical ventilation, antibiotic replacement or addition, sedation and analgesia, etc. are performed, and debridement, dressing change, and surgery are performed.

[0024] (2) Probiotic group (TP group): In addition to conventional treatment, probiotic powder (usage: Lactobacillus plantarum powder 2g, taken with warm water once a day) was given daily for 14 days; (3) Placebo group (TS group): In addition to conventional treatment, patients were given a placebo (administered with 2 g of maltodextrin, taken with warm water once a day) daily for 14 days.

[0025] A total of 61 subjects who met the inclusion criteria and completed the study were included in this study, of whom 32 were placed in the probiotic group (TP group) and the remaining 29 were placed in the placebo group (TS group). In the TP group, there were 22 males (68.800%) and 10 females (31.30%); the age was 50.50 (interquartile range (IQR) 34.50, 56.25) years; the mean body mass index (BMI) was 24.01±0.45; 24 patients were admitted to the hospital within 6 hours after injury (75.00%), 6 patients were admitted to the hospital between 6 and 12 hours after injury (18.80%), 1 patient was admitted between 12 and 24 hours after injury (3.10%), and 1 patient was admitted between 24 and 48 hours after injury (3.10%); the causes of injury were flame burns in 26 patients (81.30%), hydrothermal burns in 3 patients (9.40%), chemical burns in 2 patients (6.30%), and electric shock in 1 patient (3.10%). The total burn area was 32.00% (IQR 28.00%, 55.00%) of the total body surface area (TBSA), with third-degree burns accounting for 16.75% (IQR 10.00%, 23.50%) of the TBSA. On admission, 13 patients (40.60%) had respiratory tract burns. On admission, two patients were assessed as having a fair condition (6.30%), five as having a severe condition (15.60%), and 25 as having a critical condition (78.10%). In the TS group, there were 24 males (82.80%) and 5 females (17.20%). The average age was 49.00 (IQR 37.00, 56.00) years, with a mean BMI of 23.68±0.54. Sixteen patients were admitted to the hospital within 6 hours after injury (88.90%), 1 patient was admitted within 12-24 hours after injury (3.40%), 3 patients were admitted within 24-48 hours after injury (10.30%), and 2 patients were admitted within 24-48 hours after injury (6.90%). The causes of injury were flame burns in 21 patients (72.40%), hot fluid burns in 5 patients (17.20%), chemical burns in 1 patient (3.40%), and electric shock in 2 patients (6.90%). The total burn area was 29.00% (IQR 28.00%, 50.00%) TBSA, and the third-degree burn area was 13.00% (IQR 10.00%, 18.50%) TBSA. Nine patients (31.00%) had respiratory tract burns on admission. Eight patients (27.60%) were classified as severely ill, and 21 patients (72.40%) were classified as critically ill. Demographic characteristics of the two groups showed no statistically significant differences (P>0.05). Details are shown in Table 1.

[0026] Table 1 Comparison of baseline data between the two groups of patients 6. Baseline comparison of effect evaluation indicators The baseline data of observation indicators of the two groups of patients were compared at the time of admission. The comparison of intestinal dysfunction symptom scores was P=0.853 (P>0.05), and the intestinal dysfunction assessment scores were balanced at baseline in the two groups and were comparable. The comparison of APACHE II scores was P=0.807 (P>0.05), and the APACHE II scores were balanced at baseline in the two groups and were comparable. For details, see 2.

[0027] Table 2 Baseline data of observation indicators of the two groups of patients ( ) 7. Comparison of clinical data Comparison of clinical treatment measures other than intervention measures (including tracheotomy, mechanical ventilation, time of the first operation, number of operations within 14 days, enteral nutrition supplementation, intestinal function intervention, and use of special-grade antibiotics) between the two groups during the study revealed no statistically significant differences in these clinical treatment measures between the two groups (P>0.05). See 3 for details.

[0028] Table 3 Comparison of clinical data between the two groups of patients 8. Intestinal dysfunction symptom score The intestinal dysfunction symptoms of the two groups of patients were scored before intervention (D0) and on the 3rd, 7th and 14th days of intervention (D3, D7, D14), and statistical analysis was performed.

[0029] Intra-group comparison: Comparison of intestinal dysfunction symptom scores between the two groups: in the TP group, the difference between D3 and D0 was H=0.000, P=1.000 (P>0.05); the difference between D7 and D0 was H=1.194, P=0.033 (P<0.05), and the difference was statistically significant; the difference between D14 and D0 was H=2.250, P<0.001, and the difference was statistically significant; the difference between D7 and D3 was H=1.194, P=0.033 (P<0.05), and the difference was statistically significant; the difference between D14 and D3 was H=2.250, P<0.001, and the difference was statistically significant; the difference between D14 and D7 was H=1.056, P=0.085 (P>0.05), and the difference was not statistically significant. In the TS group, the H values for D3 compared with D0 were 0.806, P=0.367 (P>0.05); the H values for D7 compared with D0 were 0.389, P=1.000 (P>0.05); the H values for D14 compared with D0 were 1.306, P=0.014 (P<0.05), and the differences were statistically significant; the H values for D7 compared with D3 were 0.417, P=1.000 (P>0.05); the H values for D14 compared with D3 were 2.111, P<0.001, and the H values for D14 compared with D7 were 1.694, P<0.001, and the differences were statistically significant.

[0030] Comparison between groups: The intestinal dysfunction symptom scores of the two groups at different time points were compared. On D3, the comparison between the two groups was Z=1.807, P=0.074 (P>0.05); on D7, the comparison between the two groups was Z=2.008, P=0.047 (P<0.05); on D14, the comparison between the two groups was Z=2.214, P=0.031 (P<0.05). The differences between the two groups on D7 and D14 were statistically significant. See Table 4 for details. Figure 1 .

[0031] Table 4 Comparison of intestinal dysfunction symptom scores between the two groups of patients at different times [M(P25, P75)] Note: a. There were significant differences between D3 and DO (P < 0.05); b. There were significant differences between D7 and D0 for each index (P < 0.05); c. There were significant differences between D14 and D0 for each index (P < 0.05); d. There were significant differences between D7 and D3 (P < 0.05); e. There were significant differences between D14 and D3 (P < 0.05); f. There were significant differences between D14 and D7 (P < 0.05).

[0032] in Figure 1TPD0: before intervention in the TP group; TPD7: 7th day of intervention in the TP group; TPD14: 14th day of intervention in the TP group; TSD0: before intervention in the TS group; TPD7: 7th day of intervention in the TP group; TPD14: 14th day of intervention in the TS group; * indicates that there is a statistical difference between the two groups (P < 0.05), ** indicates that there is a statistical difference between the two groups (P < 0.01), and *** indicates that there is a statistical difference between the two groups (P < 0.001).

[0033] Because the intestinal dysfunction symptom scores of the two groups at different time points were not completely normally distributed, GEE analysis was performed. The overall GEE model revealed significant effects of group and time (P = 0.042 and < 0.001, respectively), indicating that both Lactobacillus plantarum supplementation and treatment duration significantly altered intestinal dysfunction symptom scores. Furthermore, there was a significant interaction effect between group and time (P = 0.016), suggesting that the temporal trends differed between the two groups. Further analysis by group revealed that in the TP group, intestinal dysfunction symptom scores showed a significant time-dependent downward trend compared with D0, with more pronounced decreases at D7 and D14 [D7: OR 0.200 (95% CI 0.073-0.543), P < 0.01; D14: OR 0.026 (95% CI 0.013-0.049), P < 0.001]. In the TS group, compared with D0, the intestinal dysfunction symptom scores increased on D3 and D7, especially on D3 [OR 5.008 (95% CI 1.780-14.093), P < 0.01]. The intestinal dysfunction symptom scores decreased significantly on D14 [OR 0.103 (95% CI 0.046-0.231), P < 0.001]. On D3, D7, and D14, the intestinal dysfunction symptom scores of patients in the TP group were significantly lower than those in the TS group [between-group differences: D3: OR 0.223 (95% CI 0.053-0.941), P < 0.05; D7: OR 0.103 (95% CI 0.014-0.763), P < 0.05; D14: OR 0.311 (95% CI 0.126-0.767), P < 0.05]. The specific results are shown in Tables 5, 6 and 7.

[0034] Table 5 GEE overall model effect of intestinal dysfunction symptom score Table 6 GEE analysis results of intestinal dysfunction symptom scores at different time points in the TP and TS groups Note: This analysis takes D0 as the baseline. The values in the table are the comparison results of the two groups at each time point with D0. Table 7 GEE analysis results of intestinal dysfunction symptom scores of the two groups of patients at different time points 9. 16S rRNA sequencing results of intestinal flora (1) Basic analysis of sequencing results This study selected 90 stool samples from the collected samples for testing based on patient compliance, strict implementation, sample retention time, method, and quantity during the study. They were divided into 6 groups according to grouping and collection time, with 15 samples in each group (i.e., 15 samples in each of the six subgroups of TPD0, TPD7, TPD14, TSD0, TSD7, and TSD14). All samples passed the amplification test before sequencing, and some amplification test results are as follows Figure 2 shown.

[0035] After sequencing, the sequencing samples that passed quality control generated a total of 4041 amplicon sequence variants (ASVs), of which 1508 ASVs were obtained in the TPD0 group, 1048 ASVs were obtained in the TPD7 group, and 1023 ASVs were obtained in the TPD14 group; 1635 ASVs were obtained in the TSD0 group, 892 ASVs were obtained in the TSD7 group, and 978 ASVs were obtained in the TSD14 group. The Venn diagram can intuitively display the number of common and unique characteristic sequences between different groups, thereby clearly reflecting the overlap of characteristics between samples. Figure 3 As shown in the figure (TPD0: before intervention of TP group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TP group; TSD0: before intervention of TS group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TS group), there were 336 ASVs in total in the three subgroups of TP group, and there were 283 ASVs in total in the three subgroups of TS group. The order of the total number of ASVs was TSD0>TPD0>TPD7>TPD14>TSD14>TSD7.

[0036] (2) Changes in α-diversity of intestinal flora ①Species dilution curve Rarefaction curves can be used to evaluate the comparability of species diversity between different groups. In this paper, using Chao1 index as the index type and ASV / OTU as the classification level, it was observed that the total number of species tended to stabilize with the increase of sequencing depth. This shows that the selection of sequencing depth is reasonable and has fully covered the species diversity in the sample. Figure 4 shown.

[0037] ②Abundance level curve The Rank Abundance Curve can reflect the distribution pattern of ASV / OTU abundance in each sample. The wider the width, the more diverse the species contained in the sample, and the flatter the curve, the more balanced the species distribution. In this study, the curve of each subgroup showed a large width and the curve gradually flattened, indicating that the species in the sample were rich and relatively evenly distributed. Figure 5 As shown (TPD0: before intervention of TP group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TP group; TSD0: before intervention of TS group; TPD7: 7th day of intervention of TP group; TPD14: 14th day of intervention of TS group).

[0038] ③α diversity index Alpha diversity measures the richness, diversity, and evenness of species within a uniform habitat, also known as within-habitat diversity. The Good's_coverage index reflects the coverage of species diversity within a sample by sequencing data. The Chao1 and Observed_species indices reflect species richness, the Shannon and Simpson indices reflect diversity, and the Pielou_e index reflects evenness. In this study, the Good's_coverage index for characterization coverage of all six subgroups was above 99.9%, indicating that sequences within the sample were largely detected. The remaining indices were then further compared.

[0039] Intra-group comparison: The α-diversity of intestinal flora was compared between the two groups at different time points. In the TP group, Chao1, Observed_species, Simpson, Shannon, and Pielou_e indices decreased on day 7 (TPD7) compared with before intervention (TPD0), with statistically significant differences in Chao1 and Observed_species (P < 0.05). On day 14 (TPD14), Chao1, Observed_species, Simpson, Shannon, and Pielou_e indices increased compared with day 7 (TPD7), but were still slightly lower than before intervention (TPD0), but the differences were not statistically significant (P > 0.05). In the TS group, the Chao1, Observed_species, Simpson, Shannon, and Pielou_e indices decreased on the 7th day of intervention (TSD7) compared with those before intervention (TSD0), and the differences in Chao1, Observed_species, Shannon, and Pielou_e indices were statistically significant (P < 0.05). On the 14th day of intervention (TSD14), the Chao1 and Observed_species indices continued to decrease, and the differences were statistically significant (P < 0.01). The other indices increased slightly, but the Shannon index still had a statistically significant difference compared with TSD0 (P < 0.05).

[0040] Intergroup comparison: Before intervention (D0), there were no statistically significant differences in the Chao1, Observed_species, Simpson, Shannon, and Pielou_e indices between the TP and TS groups (P>0.05), suggesting that the α-diversity of the intestinal microbiota was comparable between the two groups at baseline. After 7 days of intervention, the Shannon, Simpson, and Pielou_e indices were higher in the TP group than in the TS group, with statistically significant differences in the Simpson and Pielou_e indices (P<0.05). The Chao1 and Observed_species indices were slightly lower in the TP group than in the TS group, but the differences were not statistically significant (P>0.05). After 14 days of intervention, the Chao1, Observed_species, Simpson, Shannon, and Pielou_e indices were higher in the TP group than in the TS group, with statistically significant differences in the Chao1 and Observed_species indices between the two groups (P<0.05). The above results show that although the α diversity of intestinal flora decreased in both groups of patients in the early stage, the decrease was more obvious in the TS group. In the later stage of intervention, the α diversity of intestinal flora in the TP group was significantly restored, and the recovery effect of intestinal microbial α diversity in the TP group was better than that in the TS group. Figure 6 .

[0041] Table 8 Comparison of intestinal flora α diversity between the two groups of patients at different time points Note: * indicates that there is a statistical difference between the group and D0 (P < 0.05), ** indicates that there is a statistical difference between the group and D0 (P < 0.01).

[0042] (3) Changes in intestinal flora β diversity Beta diversity is used to compare the compositional differences of microbial communities between different samples, thereby assessing their similarities or differences. Beta diversity is typically measured by calculating a distance matrix between samples. Common methods include Bray-Curtis distance, Jaccard index, and UniFrac distance.

[0043] Principal coordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) were performed based on the jaccard distance matrix algorithm.

[0044] The results are as follows Figure 7 As shown (where Figure 7 A in the figure is the PCoA analysis diagram based on jaccard distance; Figure 7 Figure B shows an NMDS analysis based on the jaccard distance. In the PCoA plot, most samples in the TP and TS groups were close before the intervention (TPD0 and TSD0), and their confidence ellipses largely overlapped, indicating that the gut microbiota of the two subgroups were relatively similar. On intervention day 7 (TPD7), most samples in the TP group were farther from their pre-intervention (TPD0) locations, but on intervention day 14 (TPD14), most samples were closer to their pre-intervention (TPD0) locations, with a high degree of overlap between their confidence ellipses. In contrast, on intervention days 7 (TSD7) and 14 (TSD14), several samples in the TS group were farther from their pre-intervention (TSD0) locations, with their confidence ellipses deviating from those of pre-intervention (TSD0). The NMDS results were similar to those of the PCoA, indicating that the β-diversity of the gut microbiota in the TP group was gradually recovering, while the recovery of β-diversity in the TS group was less pronounced.

[0045] The anosim matrix test method was further used to quantitatively detect and analyze the distance between groups. The present study found that within the TP group, there were statistically significant differences between the TPD0 and TPD7 subgroups ( P <0.01), there was a statistically significant difference between TPD7 and TPD14 subgroups ( P<0.05), while there was no statistically significant difference between TPD0 and TPD14 subgroups ( P =0.186). Within the TS group, there were statistically significant differences between the TSD0, TSD7, and TSD14 subgroups ( P <0.01), there was no statistically significant difference between TSD7 and TSD14 subgroups ( P =0.359). Comparison between the two groups at different time points showed that there was no statistically significant difference between the TP group and the TS group at D0 and D7 ( P =0.367,0.400); at D14, the difference between the two groups was statistically significant ( P <0.05). Therefore, these results suggest that supplementation with Lactobacillus plantarum is beneficial for restoring the β diversity of the intestinal flora in patients. Figure 8 shown.

[0046] Table 9 Statistics of sample distance difference analysis between groups Note: TPD0: before intervention in TP group; TPD7: 7th day of intervention in TP group; TPD14: 14th day of intervention in TP group; TSD0: before intervention in TS group; TPD7: 7th day of intervention in TP group; TPD14: 14th day of intervention in TS group; “_” indicates no such value.

[0047] 10. Analysis of intestinal flora species composition (1) Analysis of species taxonomic composition First, the relative abundance distribution maps of the intestinal flora of the six groups of samples were drawn from the phylum and genus biological classification levels, and then the differences of these species with higher abundance were compared.

[0048] At the phylum level, Firmicutes ( Firmicutes ), Proteobacteria ( Proteobacteria ), Bacteroidetes ( Bacteroidetes ), Actinobacteria ( Actinobacteri a) and Verrucomicrobia ( Verrucomicrobia ) were the dominant bacterial phyla in each group of samples.

[0049] Intra-group comparisons: The top five phyla in terms of relative abundance of intestinal flora were compared between the two groups at different time points. In the TP group, the relative abundance of Firmicutes and Proteobacteria increased on day 7 (D7) compared with before intervention (D0), but the differences were not statistically significant (P>0.05). The relative abundance of Bacteroidetes, Verrucomicrobia, and Actinobacteria decreased, with a statistically significant difference in Bacteroidetes (P<0.05). On day 14 (D14), the relative abundance of Firmicutes and Actinobacteria decreased, but the differences were not statistically significant (P>0.05), while the relative abundance of Proteobacteria continued to increase, but the differences were not statistically significant (P>0.05). The relative abundance of Bacteroidetes increased compared with D7, but remained slightly lower than D0, but the differences were not statistically significant (P>0.05). The relative abundance of Verrucomicrobia increased compared with D7 and D0, but the differences were not statistically significant (P>0.05). In the TS group, the relative abundance of Firmicutes increased on the 7th day of intervention (D7) compared with that before intervention (D0), but the difference was not statistically significant (P>0.05); the relative abundance of Proteobacteria, Bacteroidetes, Actinobacteria, and Verrucomicrobia decreased, among which the difference in Proteobacteria was statistically significant (P<0.05); on the 14th day of intervention (D14), the relative abundance of Firmicutes decreased compared with D7, but was still higher than D0, but the difference was not statistically significant (P>0.05); the relative abundance of Proteobacteria increased compared with D7 and D0, but the difference was not statistically significant (P>0.05); the relative abundance of Bacteroidetes increased slightly compared with D7, but was still lower than D0, and the difference was statistically significant compared with D0 (P<0.05); the relative abundance of Actinobacteria and Verrucomicrobia continued to decrease, but the difference was not statistically significant (P>0.05).

[0050] Comparison between groups: On D0, there was no statistically significant difference in the relative abundance of the five major bacterial phyla between the two groups (P>0.05), indicating that the baseline bacterial flora composition was comparable. On D7 and D14, no statistically significant difference was found in the relative abundance of each bacterial phylum between the two groups (P>0.05). Figure 9 and Figure 10 shown.

[0051] Table 10 The top five phylum-level species with the highest relative abundance at different times in the two groups of patients From the genus level, the top 10 relative abundance genera are: Enterococcus ( Enterococcus ), Bacteroides ( Bacteroides ), Streptococcus ( Streptococcus ), Bifidobacterium spp. ( Bifidobacterium ), Parabacteroides spp. Parabacteroides ), Collinsella spp. ( Collinsella ), Lactococcus ( Lactococcus ), Akkermansia muciniphila ( Akkermansia), Lactobacillus spp. ( Lactobacillus ), Blautia ( Blautia ).

[0052] Within-group analysis: Further statistical analysis revealed that within the TP group, the relative abundance of Enterococcus and Streptococcus increased significantly on D7 (P < 0.05) and returned to baseline levels by D14 (P > 0.05 compared with D0). Parabacteroides and Blautia decreased significantly on D7 (P < 0.05). Although they did not fully return to baseline levels by D14, the differences were not statistically significant (P > 0.05). Lactococcus increased in a time-dependent manner, with a significant difference between D7 and D0 (P < 0.05). Bacteroides, Akkermansia, and Collinsella decreased on D7 and recovered on D14, but the differences within the groups were not statistically significant (P > 0.05). Bifidobacterium decreased slightly compared with D0 (P > 0.05), while Lactobacillus increased gradually, but the differences were not statistically significant (P > 0.05). In the TS group, the relative abundance of Enterococcus increased significantly on D7 (P < 0.05), then decreased on D14 compared to D7 but remained above baseline (P < 0.05). Bacteroides decreased significantly on D7 (P < 0.01), partially recovered on D14 but remained below D0 (P < 0.05). Parabacteroides and Blautia were significantly lower on both D7 and D14 than on D0 (P < 0.05). Bifidobacterium decreased sharply on D7 (P < 0.001), then recovered slightly on D14 but remained significantly below baseline (P < 0.001). Lactococcus and Lactobacillus showed a non-significant increasing trend (P > 0.05), while Akkermansia showed a gradual decrease (P > 0.05). Collinsella showed a slight fluctuation, decreasing on D7 and recovering on D14 (P > 0.05). The relative abundance of Streptococcus remained stable (P > 0.05).

[0053] Comparisons between groups: Comparison of the top 10 species in the intestinal flora at different time points between the two groups revealed no statistically significant difference in the relative abundance of the top 10 genera between the two groups at D0 (P>0.05), indicating baseline comparability. At D7, the relative abundance of Lactococcus and Lactobacillus was significantly higher in the TP group than in the TS group (P<0.05). Bifidobacterium, Akkermansia, Collinsella, Bacteroides, and Streptococcus were slightly higher in the TP group, while Enterococcus, Parabacteroides, and Blautia were slightly lower, but the differences were not statistically significant (P>0.05). At D14, the relative abundance of Enterococcus was significantly higher in the TS group than in the TP group (P<0.05), while the relative abundance of Lactococcus and Bifidobacterium was significantly higher in the TP group than in the TS group (P<0.05). The genera Lactobacillus, Akkermansia, Collinsella, and Parabacteroides were slightly higher in the TP group, but the difference was not statistically significant (P>0.05). The differences in the other genera between the groups were not significant. See Table 11 for details. Figure 11 and Figure 12 , ** indicates that there is a statistical difference between the group and D0 (P < 0.01), *** indicates that there is a statistical difference between the group and D0 (P < 0.001).

[0054] Table 11 Top 10 relative abundance species at genus level in the two groups of patients at different times (2) Analysis of species abundance distribution trends In the present invention, a cluster heat map was generated based on the average abundance of the top 20 species at the genus level to analyze the species abundance distribution trend of each group of samples. The species abundance of the TP group and the TS group before intervention (TPD0 and TSD0) were very similar. On the 7th day of intervention (TPD7), the TP group had a high abundance of Streptococcus ( Streptococcus ), Enterococcus spp. ( Enterococcus ), Actinomycetes ( Actinomyces ), Lactobacillus spp. ( Lactobacillus ), Lactococcus ( Lactococcus ), with a high abundance; on the 14th day of intervention (TPD14), Lactococcus ( Lactococcus ), Lactobacillus spp. ( Lactobacillus ), Prevotella ( Prevotella ), Akkermansia ( Akkermansia ), Morganella ( Morganella ) were more abundant. In the TS group, on the 7th day of intervention (TSD7), Enterococcus genus ( Enterococcus ), Actinomycetes ( Actinomyces ), Prevotella ( Prevotella ), Parabacteroides spp. Parabacteroides ), Streptococcus ( Streptococcus ) were more abundant; on the 14th day of intervention (TSD14), Citrobacter ( Citrobacter ), Morganella ( Morganella ), Enterococcus spp. ( Enterococcus ) abundance was higher. Overall, the species abundance distribution of the intestinal flora in the TP group after 14 days of intervention (TPD14) was closer to that before intervention (TPD0), such as Figure 13 shown.

[0055] (3) Species classification level analysis The taxonomic hierarchy tree can be used to simultaneously display the composition ratios of different taxonomic units in different groups. It can supplement the abundance data of each ASV or OTU (operational taxonomic unit) group into the graph in the form of a pie chart, and reflect the composition of microorganisms at different taxonomic levels. In the present invention, the intestinal flora of the two groups of patients at different time points showed differences in multiple taxonomic units, especially the TS group, which showed obvious differences at multiple taxonomic levels at different time points, such as Figure 14shown.

[0056] 11. Comparison of Acute Physiology and Chronic Health Evaluation (APACHE II) The two groups of patients were scored with APACHEⅡ before intervention (D0) and on the 3rd, 7th and 14th days of intervention (D3, D7 and D14) to dynamically evaluate the overall condition of the patients.

[0057] Intra-group comparison: APACHE II scores at different time points in the TP group and TS group were compared. In the TP group, the difference between D3 and D0 was not statistically significant, P=0.826 (P>0.05); the difference between D7 and D0 was statistically significant, P=0.169 (P<0.05); the difference between D14 and D0 was statistically significant, P=0.033 (P<0.05); the difference between D7 and D3 was not statistically significant, P=1.000 (P>0.05); the difference between D14 and D3 was not statistically significant, P=1.000 (P>0.05); the difference between D14 and D7 was not statistically significant, P=1.000 (P>0.05). In the TS group, there was no statistically significant difference between D3 and D0, P=1.000 (P>0.05); there was no statistically significant difference between D7 and D0, P=1.000 (P>0.05); there was a statistically significant difference between D14 and D0, P=0.102 (P<0.05); there was no statistically significant difference between D7 and D3, P=0.272 (P>0.05); there was no statistically significant difference between D14 and D3, P=0.014 (P<0.05); there was no statistically significant difference between D14 and D7, P=1.000 (P>0.05).

[0058] Comparison between groups: APACHE II scores at different time points in the TP group and the TS group were compared, with P>0.05, and the difference was not statistically significant. Figure 15 .

[0059] Table 12 Comparison of APACHE II scores between the two groups at different time points Note: a. There were differences between D3 and DO (P < 0.05); b. There were differences between D7 and D0 for each index (P < 0.05); c. There were differences between D14 and D0 for each index; d. There were differences between D7 and D3 (P < 0.05); e. There were differences between D14 and D3 (P < 0.05); f. There were differences between D14 and D7 (P < 0.05).

[0060] Because the APACHE II scores of the two groups of patients at different time points were not completely normally distributed, generalized estimating equations (GEE) were used for analysis. Overall, the group effect of APACHE II scores in the two groups was not significant (P=0.415); the time effect was significant (P<0.001), indicating that the APACHE II score did not decrease significantly after supplementation with Lactobacillus plantarum, but treatment time had a significant effect on the decrease in APACHE II score; in addition, the interaction effect between the groups and time was not significant (P=0.135); further analysis found that compared with the TS group, the TP group [OR 0.535 (95%CI 0.119-2.407), P>0.05], suggesting that supplementation with Lactobacillus plantarum may be a protective factor for the decrease in APACHE II score, although there was no statistical difference; at the same time, during the intervention, the APACHE II score gradually decreased on days 3, 7, and 14 [D3: OR 0.717 (95%CI 0.296-1.735), P>0.05; D7: OR 0.338 (95%CI The OR for D14 was 0.189 (95% CI 0.079-0.453), P < 0.001, indicating that the APACHE II score did not change significantly in the short term, but decreased significantly with prolonged treatment. The specific results are shown in Tables 13 and 14.

[0061] Table 13 APACHE II score GEE overall model effect Table 14 APACHE II score GEE results 12. Comparison of wound healing rates At the end of the intervention, i.e., on day 14 (D14), the wound healing rates of the two groups were compared. The average wound healing rate of the TP group was 52.01% ± 4.35%, and the average wound healing rate of the TS group was 49.93% ± 3.93%. There was no statistically significant difference between the two groups (P = 0.724). The specific results are shown in Table 15 and Figure 16 shown.

[0062] Table 15 Comparison of wound healing rates between the two groups on day 14 The embodiments of the present application are described above in conjunction with the accompanying drawings. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the core idea of the present application, but the present application is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present application, ordinary technicians in this field can also make many forms without departing from the scope of protection of the purpose of the present application and the claims, all of which are within the protection of the present application.

Claims

1. Use of a strain of Lactobacillus plantarum chen-02 in the preparation of a medicament for improving intestinal dysfunction caused by burns, characterized in that: The Lactobacillus plantarum ( Lactobacillus plantarum )chen-02 was deposited on September 13, 2021 at the General Microbiology Center of China Culture Collection Administration, located at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC NO.23397.

2. A biological agent, characterized in that The active ingredients of the biological preparation include Lactobacillus plantarum chen-02, which was deposited in the General Microbiology Center of the China Culture Collection Administration on September 13, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCC NO.23397.

3. A drug for improving intestinal dysfunction caused by burns, characterized in that: The drug includes Lactobacillus plantarum chen-02 or the biological preparation described in claim 2. The Lactobacillus plantarum chen-02 has been deposited in the General Microbiology Center of the China Culture Collection Administration on September 13, 2021, with the address at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.23397.

4. The drug according to claim 3, characterized in that The medicine also includes pharmaceutical excipients.

5. The drug according to claim 4, characterized in that The pharmaceutical excipient is at least one of water, lactose, sodium chloride and glucose.

6. The drug according to any one of claims 3 to 5, characterized in that The dosage form of the medicine is powder, granule, capsule or tablet.

7. The drug according to claim 6, characterized in that The medicine is an oral medicine.

8. Use of a strain of Lactobacillus plantarum chen-02 in the preparation of a product for improving and / or treating burn wound healing, characterized in that: The plant lactobacillus chen-02 was deposited in the General Microbiology Center of the China Culture Collection Administration on September 13, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCCNO.23397.

9. The use according to claim 8, characterized in that The product includes a drug or a biologic.

10. A drug for improving and / or treating burn wound healing, characterized in that: The drug includes Lactobacillus plantarum chen-02, which was deposited in the General Microbiology Center of the China Culture Collection Administration on September 13, 2021, at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, with the deposit number CGMCCNO.23397.

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