1.Regulation of lysosome function by stem cells in treatment of lysosomal storage diseases
Yiwen LI ; Feixiang LIU ; Yunke ZHANG
Chinese Journal of Tissue Engineering Research 2026;30(1):145-152
BACKGROUND:Lysosomal storage diseases,as a group of rare genetic metabolic disorders,exhibit complex pathogenesis often leading to dysfunction of cells,tissues,and organs.Current therapeutic approaches have certain limitations.Stem cell transplantation,as an emerging treatment method,offers new options for patients with lysosomal storage diseases.OBJECTIVE:To review the mechanisms of action of stem cells in regulating lysosomes for the treatment of lysosomal storage diseases and explore the feasibility of traditional Chinese medicine in treating such diseases,providing new insights for the treatment of lysosomal storage diseases with stem cells.METHODS:Relevant literature from 2010 to 2024 was searched in CNKI and PubMed databases using keywords"stem cells,lysosomal storage disease,lysosome"in English and Chinese.Ultimately,78 articles were included for review and analysis.RESULTS AND CONCLUSION:(1)Stem cells treat lysosomal storage diseases by regulating lysosomes primarily through three aspects:regulating stem cell differentiation and replacement,improving intercellular communication and the microenvironment,and enhancing lysosomal enzyme expression through gene editing.(2)Stem cells have achieved significant effects in the treatment of some lysosomal storage diseases,such as Niemann-Pick disease,mucopolysaccharidoses,Gaucher disease,and metachromatic leukodystrophy.(3)The procedure for stem cell transplantation needs further optimization.Adverse reactions post-transplantation urgently need to be addressed,and the efficiency and safety of gene-modified stem cells also need to be further improved.In the future,more research on the treatment of lysosomal storage diseases with traditional Chinese medicine is required to reveal the relevant mechanisms for the treatment of lysosomal storage diseases with traditional Chinese medicine.
2.Regulation of lysosome function by stem cells in treatment of lysosomal storage diseases
Yiwen LI ; Feixiang LIU ; Yunke ZHANG
Chinese Journal of Tissue Engineering Research 2026;30(1):145-152
BACKGROUND:Lysosomal storage diseases,as a group of rare genetic metabolic disorders,exhibit complex pathogenesis often leading to dysfunction of cells,tissues,and organs.Current therapeutic approaches have certain limitations.Stem cell transplantation,as an emerging treatment method,offers new options for patients with lysosomal storage diseases.OBJECTIVE:To review the mechanisms of action of stem cells in regulating lysosomes for the treatment of lysosomal storage diseases and explore the feasibility of traditional Chinese medicine in treating such diseases,providing new insights for the treatment of lysosomal storage diseases with stem cells.METHODS:Relevant literature from 2010 to 2024 was searched in CNKI and PubMed databases using keywords"stem cells,lysosomal storage disease,lysosome"in English and Chinese.Ultimately,78 articles were included for review and analysis.RESULTS AND CONCLUSION:(1)Stem cells treat lysosomal storage diseases by regulating lysosomes primarily through three aspects:regulating stem cell differentiation and replacement,improving intercellular communication and the microenvironment,and enhancing lysosomal enzyme expression through gene editing.(2)Stem cells have achieved significant effects in the treatment of some lysosomal storage diseases,such as Niemann-Pick disease,mucopolysaccharidoses,Gaucher disease,and metachromatic leukodystrophy.(3)The procedure for stem cell transplantation needs further optimization.Adverse reactions post-transplantation urgently need to be addressed,and the efficiency and safety of gene-modified stem cells also need to be further improved.In the future,more research on the treatment of lysosomal storage diseases with traditional Chinese medicine is required to reveal the relevant mechanisms for the treatment of lysosomal storage diseases with traditional Chinese medicine.
3.Timing of Termination and Cost-Effectiveness Analysis of Acupuncture for Acute Peripheral Facial Paralysis:A Randomized Controlled Trial
Xiaohan ZHANG ; Tao WANG ; Jinbo WANG ; Yiwen MIAO ; Lijuan DAI ; Jiaying ZHANG ; Shulan WANG ; Hui WANG ; Guoxin WANG ; Yuhang CHEN ; Xinjun WANG ; Bingguo XU
Journal of Traditional Chinese Medicine 2026;67(11):1185-1191
ObjectiveTo investigate the optimal termination time for acupuncture in treating patients with acute peripheral facial paralysis and its cost-effectiveness. MethodsA total of 120 eligible patients with acute-stage peri-pheral facial paralysis were randomly assigned to either the mild dysfunction termination group and the complete recovery termination group, with 60 patients in each group. Both groups received the standard acupuncture treatment protocol. Treatment in the mild dysfunction termination group was terminated when the Sunnybrook facial grade scale (SFGS) score first reached or exceeded 83 points, while that in the complete recovery termination group was terminated when the SFGS score first reached or exceeded 95 points. Assessments were conducted before treatment, 6 and 12 months after onset. SFGS, facial disability index (FDI) including physical function (FDIp) and social function (FDIs), self-rating anxiety scale (SAS), and self-rating depression scale (SDS) scores were assessed before treatment, and 6 and 12 months after onset. Any acupuncture-related adverse events during treatment were recorded for safety evaluation. Treatment sessions and medical costs including direct costs, indirect costs, insurance coverage, total societal costs, and patient out-of-pocket expenses were also recorded, and an economic evaluation was conducted including cost-effectiveness ratio (CER) and incremental cost-effectiveness ratio (ICER). ResultsUltimately, 56 patients in the mild dysfunction termination group and 55 in the complete recovery termination group completed the follow-up. At 6 and 12 months after onset, SFGS and FDIp scores in both groups improved significantly while FDIs, SAS and SDS scores decreased (P<0.05). Comparison of scores between groups 6 months and 12 months after onset showed no statistically significant differences (P>0.05). During the trial, the incidence of adverse events was 13.3% (8/60) in the mild dysfunction termination group and 18.3% (11/60) in the complete recovery termination group, with no statistically significant difference (P>0.05). The number of treatment sessions, total social costs, and out-of-pocket expenses in the mild dysfunction termination group were significantly lower than those in the complete recovery termination group (P<0.05). The CER of the mild dysfunction termination group in SFGS, FDIp, FDIs, SAS, and SDS scores was lower than that of the complete recovery termination group. The ICER analysis showed that continuing treatment until full recovery incurred an additional cost of 573.30 CNY/point in SFGS improvement, whereas 1-point improvement in FDIp, FDIs, SAS, and SDS required 21,355.25 CNY, 1779.60 CNY, 3713.96 CNY, and 2755.52 CNY, respectively. ConclusionFor acupuncture in treating acute peripheral facial palsy, terminating treatment when mild dysfunction is achieved yields long-term efficacy comparable to that of continuing treatment until complete recovery, while significantly reducing medical costs and socioeconomic burden.
4.Clinical Observation of Xiere Xingpi Yin in Treatment of Simple Obesity and Effect on Obesity-related miRNAs
Yelin ZHU ; Yiwen HUANG ; Ke ZHANG ; Xianchang ZENG ; Zhenxiu LIU ; Hejin LAI ; Feng TAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):177-185
ObjectiveTo observe the effects of Xiere Xingpi Yin (XRXPY) on the body mass and body fat distribution in patients with simple obesity, and to preliminarily explore whether the mechanism involves the regulation of specific obesity-related microRNAs (miRNAs). MethodsA randomized controlled trial design was employed to recruit patients with simple obesity diagnosed with the traditional Chinese medicine (TCM) syndrome of dampness-heat accumulating in the spleen. Using a randomized controlled trial design, 95 patients with simple obesity of dampness-heat encumbering the Spleen syndrome were recruited, after 3 were excluded, 92 were randomly assigned in a 1∶1 ratio by means of a random number table. On the basis of lifestyle intervention, the observation group received XRXPY granules, while the control group received a placebo. Serum samples and TCM symptom scores were collected before and after the 12-week observation period. Differences in body mass loss, body composition, lipid metabolism, and TCM symptom scores were compared between the groups. An obesity-related miRNA dataset was constructed by searching the Human MicroRNA Disease Database. Library construction, sequencing, and quality control were performed on 6 selected patients from each group to screen the differentially expressed miRNAs. The intersection of these miRNAs with the obesity-related miRNAs was verified by bioinformatics and correlation analysis. ResultsBaseline characteristics were comparable between the two groups before treatment. A total of 86 patients completed the study (42 cases in the observation group and 44 cases in the control group). The proportion of patients achieving ≥5% body weight reduction was significantly higher in the observation group than in the control group 81.0% (34/42) vs. 43.2% (19/44) (χ2=12.963, P<0.01). Compared with baseline within the observation group, body weight, waist circumference, and TCM syndrome score were significantly decreased (P<0.01), as were body fat percentage (P<0.01) and serum triglyceride level (P<0.05). Compared with the control group after treatment, waist circumference and TCM syndrome score were significantly lower in the observation group (P<0.01), and body weight showed a trend toward reduction. When changes from baseline were compared between the two groups, the magnitudes of decrease in body weight, waist circumference, and TCM syndrome score were all significantly greater in the observation group than in the control group (P<0.01). With respect to safety, no serious adverse events occurred in either group and no participant discontinued the study because of adverse events. Compared with baseline, alanine aminotransferase levels decreased significantly in both the control and observation groups (P<0.01), and urea decreased significantly in the observation group (P<0.01), indicating a favorable safety profile. Sequencing identified nine differentially expressed miRNAs. After intersecting with the obesity-related miRNA dataset (187 miRNAs), 3 miRNAs were identified: miR-221 (log2FC=-6.62, P=0.009), miR-29a (log2FC=6.33, P=0.016), and miR-10b (log2FC=-5.91, P=0.029). A total of 262 target genes of the 3 miRNAs were identified, which were significantly enriched in 67 pathways, including the lysine degradation pathway (P=0.001). Correlation analysis revealed a positive correlation between miR-221 and triglycerides (r=0.46, P=0.027), a negative correlation between miR-29a and waist-to-hip ratio (r=-0.41, P=0.047), and positive correlations of miR-10b with body fat percentage (r=0.49, P=0.014) and waist circumference (r=0.44, P=0.032). ConclusionXRXPY combined with lifestyle intervention can effectively reduce the body mass and improve body composition. The underlying mechanism may involve the regulation of the lysine degradation pathway via miR-221, miR-29a, and miR-10b, though further verification is required.
5.Clinical Observation of Xiere Xingpi Yin in Treatment of Simple Obesity and Effect on Obesity-related miRNAs
Yelin ZHU ; Yiwen HUANG ; Ke ZHANG ; Xianchang ZENG ; Zhenxiu LIU ; Hejin LAI ; Feng TAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):177-185
ObjectiveTo observe the effects of Xiere Xingpi Yin (XRXPY) on the body mass and body fat distribution in patients with simple obesity, and to preliminarily explore whether the mechanism involves the regulation of specific obesity-related microRNAs (miRNAs). MethodsA randomized controlled trial design was employed to recruit patients with simple obesity diagnosed with the traditional Chinese medicine (TCM) syndrome of dampness-heat accumulating in the spleen. Using a randomized controlled trial design, 95 patients with simple obesity of dampness-heat encumbering the Spleen syndrome were recruited, after 3 were excluded, 92 were randomly assigned in a 1∶1 ratio by means of a random number table. On the basis of lifestyle intervention, the observation group received XRXPY granules, while the control group received a placebo. Serum samples and TCM symptom scores were collected before and after the 12-week observation period. Differences in body mass loss, body composition, lipid metabolism, and TCM symptom scores were compared between the groups. An obesity-related miRNA dataset was constructed by searching the Human MicroRNA Disease Database. Library construction, sequencing, and quality control were performed on 6 selected patients from each group to screen the differentially expressed miRNAs. The intersection of these miRNAs with the obesity-related miRNAs was verified by bioinformatics and correlation analysis. ResultsBaseline characteristics were comparable between the two groups before treatment. A total of 86 patients completed the study (42 cases in the observation group and 44 cases in the control group). The proportion of patients achieving ≥5% body weight reduction was significantly higher in the observation group than in the control group 81.0% (34/42) vs. 43.2% (19/44) (χ2=12.963, P<0.01). Compared with baseline within the observation group, body weight, waist circumference, and TCM syndrome score were significantly decreased (P<0.01), as were body fat percentage (P<0.01) and serum triglyceride level (P<0.05). Compared with the control group after treatment, waist circumference and TCM syndrome score were significantly lower in the observation group (P<0.01), and body weight showed a trend toward reduction. When changes from baseline were compared between the two groups, the magnitudes of decrease in body weight, waist circumference, and TCM syndrome score were all significantly greater in the observation group than in the control group (P<0.01). With respect to safety, no serious adverse events occurred in either group and no participant discontinued the study because of adverse events. Compared with baseline, alanine aminotransferase levels decreased significantly in both the control and observation groups (P<0.01), and urea decreased significantly in the observation group (P<0.01), indicating a favorable safety profile. Sequencing identified nine differentially expressed miRNAs. After intersecting with the obesity-related miRNA dataset (187 miRNAs), 3 miRNAs were identified: miR-221 (log2FC=-6.62, P=0.009), miR-29a (log2FC=6.33, P=0.016), and miR-10b (log2FC=-5.91, P=0.029). A total of 262 target genes of the 3 miRNAs were identified, which were significantly enriched in 67 pathways, including the lysine degradation pathway (P=0.001). Correlation analysis revealed a positive correlation between miR-221 and triglycerides (r=0.46, P=0.027), a negative correlation between miR-29a and waist-to-hip ratio (r=-0.41, P=0.047), and positive correlations of miR-10b with body fat percentage (r=0.49, P=0.014) and waist circumference (r=0.44, P=0.032). ConclusionXRXPY combined with lifestyle intervention can effectively reduce the body mass and improve body composition. The underlying mechanism may involve the regulation of the lysine degradation pathway via miR-221, miR-29a, and miR-10b, though further verification is required.
6.Mechanism of postoperative abdominal adhesion formation and therapeutic prospect of mesenchymal stem cell exosomes
Xingzhou ZHANG ; Ming WEI ; Guoqiang DONG ; Wei DU ; Yiwen LUO ; Nan ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(1):147-155
BACKGROUND:The formation of postoperative abdominal adhesions is a complicated process,and the prevention of postoperative adhesions is an urgent problem in clinic. OBJECTIVE:To analyze the mechanism of adhesion at cellular and molecular levels,and to provide theoretical basis for the prevention and treatment of adhesion by mesenchymal stem cell exosomes. METHODS:"Abdominal adhesion,pelvic adhesion,postoperative adhesion,epithelial mesenchymal transformation,mesenchymal stem cells,stem cell exosomes,mesenchymal stem cell exosomes"were selected as Chinese and English search terms.We searched PubMed,CNKI,and Chinese biomedical literature and screened relevant articles on postoperative abdominal adhesion and mesenchymal stem cell exosomal intervention published from inception to August 2023.After systematic analysis,54 articles were finally included for the review. RESULTS AND CONCLUSION:(1)Any pathological factors such as peritoneal inflammation,mechanical injury,tissue ischemia,and foreign body implantation cause peritoneal surface injury,resulting in postoperative abdominal adhesion.The formation process of adhesion includes the interaction of peritoneal mesothelial cell repair,inflammatory response,fibrinolytic system,coagulation pathway and other processes,involving a variety of cytokines and signaling pathways.Wnt/β-catenin pathway can induce fibrosis and angiogenesis,and cooperate with transforming growth factor-β/Smads signaling pathway to stimulate fibroblast proliferation and cause peritoneal fibrosis.Meanwhile,nuclear factor-κB signaling pathway up-regulates the expression of cellular inflammatory factors,promotes fibroblast proliferation,and plays a key role in the process of tissue fibrosis.(2)The paracrine function of stem cells is an important direction of molecular intervention in abdominal adhesions based on regenerative medicine.It can participate in a variety of complex cytokines and signaling pathways involved in abdominal adhesions.(3)Compared with traditional methods for treating abdominal adhesions,mesenchymal stem cell exosome has biological activity and is safe to use.Mesenchymal stem cell exosomes without special culture and expansion have lower immunogenicity,longer stability and other advantages,can guide a normal repair and healing through a variety of ways.(4)Mesenchymal stem cell exosome has been proven to be involved in regulating the above processes of adhesion formation in previous studies,showing potential application prospects in clinical studies.However,further clinical studies are needed to explore appropriate treatment options for mesenchymal stem cell exosomes to address the problem of clinical translation.
7.Spinal curvature abnormalities and related factors among primary and secondary school students in Guangxi in 2023
LUO Yuemei, LI Yan, REN Yiwen, DONG Yonghui, CHEN Li, ZHANG Dengcheng, ZHANG Yi, MA Jun, DONG Yanhui
Chinese Journal of School Health 2025;46(5):712-716
Objective:
To investigate the prevalence and associated factors of spinal curvature abnormalities among primary and secondary school students in the Guangxi Zhuang Autonomous Region, so as to provide a scientific basis for the prevention and control of such abnormalities.
Methods:
From September to November 2023, adopting a stratified cluster random sampling method, spinal curvature screenings and questionnaire surveys were conducted among 168 931 students from grade 4 of primary school to grade 12 of high school in 111 districts and counties across 14 cities in Guangxi. Chi square tests and binary Logistic regression analysis were used to analyze influencing factors of spinal curvature abnormalities.
Results:
In 2023, the detection rate of poor posture among students above grade 4 in Guangxi was 4.24% , and the detection rate of spinal curvature abnormalities was 2.13%. The detection rate was higher among urban students (2.84%) than rural students (1.66%), boarding students (2.61%) than non-boarding students (1.60%), and high school students (3.16%) than junior high (2.45%) and primary school students (1.15%), and the differences were statistically significant ( χ 2=269.85, 221.44, 565.10, P <0.01). A trend of increasing detection rates with higher grade levels was observed ( χ 2 trend =617.63, P <0.01). Binary Logistic regression analysis indicated that students without boarding at school ( OR =0.82, 95% CI =0.75-0.90), engaging in high-intensity physical activity for over 60 min per day ≥5 days per week ( OR =0.90, 95% CI =0.82-0.98), and adequate sleep ( OR =0.87, 95% CI =0.81-0.94) had lower risks of detecting spinal curvature abnormalities ( P <0.05).
Conclusions
The prevalence of spinal curvature abnormalities increases with grade level among primary and secondary school students in Guangxi. Regular moderate-to-vigorous physical activity demonstrates protective effects against spinal abnormalities.
8.Establishment and evaluation of a machine learning prediction model for sepsis-related encephalopathy in the elderly.
Xiao YUE ; Yiwen WANG ; Zhifang LI ; Lei WANG ; Li HUANG ; Shuo WANG ; Yiming HOU ; Shu ZHANG ; Zhengbin WANG
Chinese Critical Care Medicine 2025;37(10):937-943
OBJECTIVE:
To construct machine learning prediction model for sepsis-associated encephalopathy (SAE), and analyze the application value of the model on early identification of SAE risk in elderly septic patients.
METHODS:
Patients aged over 60 years with a primary diagnosis of sepsis admitted to intensive care unit (ICU) from 2008 to 2023 were selected from Medical Information Mart for Intensive Care-IV 2.2 (MIMIC-IV 2.2). Demographic variables, disease severity scores, comorbidities, interventions, laboratory indicators, and hospitalization details were collected. Key factors associated with SAE were identified using univariate Logistic regression analysis. The data were randomly divided into training and validation sets in a 7 : 3 ratio. Multivariable Logistic regression analysis was conducted in the training set and visualized using a nomogram model for prediction of SAE. The discrimination of the model was evaluated in the validation set using the receiver operator characteristic curve (ROC curve), and its calibration was assessed using calibration curve. Furthermore, multiple machine learning algorithms, including multi-layer perceptron (MLP), support vector machine (SVM), naive bayes (NB), gradient boosting machine (GBM), random forest (RF), and extreme gradient boosting (XGB), were constructed in the training set. Their predictive performance was subsequently evaluated on the validation set. Taking the XGB model as an example, the interpretability of the model through the SHapley Additive exPlanations (SHAP) algorithm was enhanced to identify the key predictive factors and their contributions.
RESULTS:
A total of 2 204 septic patients were finally enrolled, of whom 840 developed SAE (38.1%). A total of 21 variables associated with SAE were screened through univariate Logistic regression analysis. Multivariable Logistic regression analysis showed that endotracheal intubation [odds ratio (OR) = 0.40, 95% confidence interval (95%CI) was 0.19-0.88, P < 0.001], oxygen therapy (OR = 0.76, 95%CI was 0.53-0.95, P = 0.023), tracheotomy (OR = 0.20, 95%CI was 0.07-0.53, P < 0.001), continuous renal replacement therapy (CRRT; OR = 0.32, 95%CI was 0.15-0.70, P < 0.001), cerebrovascular disease (OR = 0.31, 95%CI was 0.16-0.60, P < 0.001), rheumatic disease (OR = 0.44, 95%CI was 0.19-0.99, P < 0.001), male (OR = 0.68, 95%CI was 0.54-0.86, P = 0.001), and maximum anion gap (AG; OR = 0.95, 95%CI was 0.93-0.97, P < 0.001) were associated with an decreased probability of SAE, and age (OR = 1.05, 95%CI was 1.03-1.06, P < 0.001), acute physiology score III (APSIII; OR = 1.02, 95%CI was 1.01-1.02, P < 0.001), Oxford acute severity of illness score (OASIS; OR = 1.04, 95%CI was 1.03-1.06, P < 0.001), and length of hospital stay (OR = 1.01, 95%CI was 1.01-1.02, P < 0.001) were associated with an increased probability of SAE. A nomogram model was constructed based on these variables. In the validation set, ROC curve analysis showed that the model achieved an area under the ROC curve (AUC) of 0.723, and the calibration curve showed good consistency between the predicted probability of the model and the observed probability. Among the machine learning algorithms, including MLP, SVM, NB, GBM, RF, and XGB, the SVM model and RF model demonstrated relatively good predictive performance, with AUC of 0.748 and 0.739, respectively, and the sensitivity was both exceeding 85%. The predictive performance of the XGB model was explained through SHAP analysis, and the results indicated that APSIII score (SHAP value was 0.871), age (SHAP value was 0.521), and OASIS score (SHAP value was 0.443) were important factors affecting the predictive performance of the model.
CONCLUSIONS
The machine learning-based SAE prediction model exhibits good predictive capability and holds significant application value for the early identification of SAE risk in elderly septic patients.
Humans
;
Machine Learning
;
Aged
;
Sepsis-Associated Encephalopathy
;
Sepsis/complications*
;
Intensive Care Units
;
Logistic Models
;
Middle Aged
;
Male
;
ROC Curve
;
Female
;
Bayes Theorem
;
Nomograms
;
Support Vector Machine
;
Algorithms
9.Mining of key genes for xylose metabolism and cloning, expression, and enzymatic characterization of XylA in Bacillus coagulans.
Yiwen ZHANG ; Yajie ZHANG ; Manxin CHEN ; Xiaojun GUO ; Baocheng ZHU ; Yuming ZHANG
Chinese Journal of Biotechnology 2025;41(10):3876-3890
Bacillus coagulans can utilize the hydrolyzed carbon source of agricultural waste to produce lactic acid via a homofermentative pathway. However, a significant carbon source metabolic repression effect was observed when the strain metabolized mixed sugars (glucose and xylose), reducing the productivity of lactic acid. In this study, we obtained the fermentation conditions for the simultaneous utilization of the mixed sugars by B. coagulans by changing the ratio of glucose to xylose in the medium. Through transcriptome sequencing, several key genes responsible for xylose utilization were identified. The critical role of xylose isomerase (XylA, EC 5.3.1.5) in the synchronous utilization of glucose/xylose in B. coagulans was investigated via qRT-PCR (quantitative real-time polymerase chain reaction). Subsequently, the heterologous expression and characterization of the XylA-encoding gene (XylA) were conducted. It was determined that the gene encoded a protein composed of 440 amino acid residues. The secondary structure of the encoded protein was predominantly composed of α-helixes and random coils, while the higher structure of the protein was identified as a homotetramer. Then, XylA was cloned and expressed in Escherichia coli BL21(DE3), and the recombinant protein Bc-XlyA was obtained with a molecular weight of approximately 50 kDa. The optimal pH and temperature of Bc-XylA were 8.0 and 60 ℃, respectively, and Mn2+, Mg2+, and Co2+ had positive effects on the activity of Bc-XlyA. The present study provides scientific data on the molecular modification of B. coagulans, offering theoretical support for the efficient utilization of xylose in the strain.
Xylose/metabolism*
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Cloning, Molecular
;
Bacillus coagulans/enzymology*
;
Aldose-Ketose Isomerases/metabolism*
;
Fermentation
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Bacterial Proteins/metabolism*
;
Glucose/metabolism*
10.Protective role of self-assembled nanoparticle vaccine of Pseudomonas aeruginosa in a mouse model of bronchiectasis with acute infection
Ziyu WU ; Yueyue ZHANG ; Yiwen ZHANG ; Jinqiong YAN ; Zifan ZHU ; Meilin WU ; Yating WANG ; Hongrong CUI ; Jiang GU ; Ying WANG ; Quanming ZOU
Journal of Army Medical University 2025;47(10):1049-1058
Objective To establish a mouse model of bronchiectasis with acute infection and evaluate the immunogenicity and protective effect of a self-assembling Pseudomonas aeruginosa(PA)nanoparticle vaccine rePO-FN based on fusion of PcrV-OprI(rePO)protein with self-assembling ferritin(Ferritin).Methods ① SPF-grade female C57BL/6 mice(aged 6~8 weeks,weighing 18~20 g)were randomly allocated into normal saline group,and low-,medium-and high-dose elastase groups(n=6).A mouse model of bronchiectasis was established via intratracheal instillation of different doses of elastase(30 μL of normal saline containing 0.65,1.30 and 2.60 IU elastase)for 3 consecutive days.At 14 and 21 d after modeling,ELISA and HE staining were performed respectively to detect the concentration of IL-6 and to observe pathological changes in lung tissue in order to confirm the modeling.② A recombinant plasmid encoding the gene of fusion protein rePO-FN was constructed and expressed in E.coli.The target protein was purified via affinity chromatography and renatured to obtain the desired protein.The physicochemical properties of the rePO-FN protein were characterized using SDS-PAGE protein gel electrophoresis,dynamic light scattering,molecular sieve chromatography,and transmission electron microscopy.③ C57BL/6J mice were randomly divided into PBS group,rePO group,rePO-FN group,and Ferritin group(n=10).The mice in the above groups were immunized intramuscularly with 100 μL PBS buffer alone or containing 10 μg of corresponding proteins on days 0,7,and 14.ELISA was used to measure the specific antibodies in serum.In 7 d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the PBS,rePO,and rePO-FN groups.After establishing a bronchiectasis model by intratracheal instillation of 2.60 IU of elastase in C57BL/6J mice as described above,the mice were randomly divided into bronchiectasis PBS group,bronchiectasis rePO group,and bronchiectasis rePO-FN group(n=10).Immunization was conducted at the same dose and procedure as described above,in 21 d after bronchiectasis modeling.At the 7th d after the final immunization,an acute PA infection model was used to compare the survival rates and bacterial colonization among the groups.Results ①Repeated intratracheal instillation of elastase significantly increased the concentration of IL-6 in the lung tissue when compared to the content of the normal saline group(P<0.05).Pathological observations revealed varying degrees of bronchial wall destruction,alveolar fusion,edema,neutrophil infiltration,and hemorrhage,with the severity increasing with elastase dose,which confirming successful establishment of the mouse model of bronchiectasis.② Well-dispersed rePO-FN nanoparticles were successfully prepared,with an average particle size of 91.28 nm,a Zeta potential of approximately-6.5 mV,and a polydispersity index(PDI)of 0.306.Molecular sieve chromatography determined the elution volume of rePO-FN protein to be 8.80 mL,corresponding to a molecular weight of approximately 1 400 kDa.③ Under acute PA XN-1 strain infection,the survival rate of the rePO-FN immunization group and the bronchiectasis rePO-FN immunization group were significantly higher than that of the PBS control group(P<0.05).Additionally,bacterial colonization in the lung tissues was significantly lower in the rePO-FN immune group and the bronchiectasis rePO-FN immune group under acute PA XN-1 strain infection than that in the rePO group and the bronchiectasis rePO group(P<0.05).Conclusion Our vaccine rePO-FN can effectively trigger a strong humoral immune response and provide significant protection against PA infection in a mouse bronchiectasis model.


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