1.The effect of bovine milk-derived extracellular vesicles on the biofilm of staphylo-coccus aureus
Ziqiang GAO ; Zi WANG ; Peng LIU ; Yangyang SONG ; Xiaolin LI ; Huaxue SONG ; Binglei SHEN
Chinese Journal of Veterinary Science 2025;45(11):2430-2438
The objective of this experiment was to investigate the inhibitory effect and mechanism of mammary-derived extracellular vesicles(MmEVs)from mastitis dairy cows on the biofilm for-mation of Staphylococcus aureus SA1.The biofilm-forming ability of Staphylococcus aureus SA1 was confirmed using Congo red staining,and the biofilm growth curve of S.aureus SA1 was plot-ted using the crystal violet staining method.The minimum inhibitory concentration(MIC)and minimum biofilm inhibitory concentration(MBIC)of MmEVs against S.aureus SA1 were deter-mined.After treating S.aureus SA1 with different concentrations of MmEVs,the cell morphology of S.aureus SA1 was observed using transmission electron microscopy.The effects of MmEVs on S.aureus SA1 under low pH(pH value=5)or heat stress(58℃)were investigated.The hydro-phobicity index was explored using the microbial adhesion to hydrocarbons(MATH)assay.Bacte-rial conductivity was measured.The expression levels of biofilm-related genes(SarA,icaB,FnbA,ClfB,CidA,and gyrB)were detected using quantitative real-time PCR(qPCR).The results showed that MIC of MmEVs against the biofilm of S.aureus SA1 was 1 000 mg/L,and the MBIC was 500 mg/L.Under the influence of MmEVs,the internal substances of S.aureus SA1 leaked,the biofilm boundary became blurred,and the cell wall separated.At the MBIC concentration,MmEVs significantly reduced the tolerance of S.aureus SA1 to low pH(P<0.001)and high tem-perature(P<0.001),decreased hydrophobicity(P<0.001),and increased bacterial conductivity(P<0.001).At the MBIC concentration,MmEVs significantly downregulated the gene expression of Sa rA(P<0.001),icaB(P<0.001),FnbA(P<0.001),ClfB(P<0.001),and CidA(P<0.001)in S.aureus SA1,while no significant effect was observed on the expression of the gyrB gene.In summary,MmEVs inhibit the formation of Staphylococcus aureus SA1 biofilms by sup-pressing the gene expression of SarA,icaB,FnbA,ClfB,and CidA within the biofilm.This dis-ruption damages the biofilm's morphological structure,reduces its tolerance to low pH and high temperature,decreases hydrophobicity,and increases bacterial conductivity,thereby ultimately in-hibiting the formation of S.aureus SA1 biofilms.
2.The effect of bovine milk-derived extracellular vesicles on the biofilm of staphylo-coccus aureus
Ziqiang GAO ; Zi WANG ; Peng LIU ; Yangyang SONG ; Xiaolin LI ; Huaxue SONG ; Binglei SHEN
Chinese Journal of Veterinary Science 2025;45(11):2430-2438
The objective of this experiment was to investigate the inhibitory effect and mechanism of mammary-derived extracellular vesicles(MmEVs)from mastitis dairy cows on the biofilm for-mation of Staphylococcus aureus SA1.The biofilm-forming ability of Staphylococcus aureus SA1 was confirmed using Congo red staining,and the biofilm growth curve of S.aureus SA1 was plot-ted using the crystal violet staining method.The minimum inhibitory concentration(MIC)and minimum biofilm inhibitory concentration(MBIC)of MmEVs against S.aureus SA1 were deter-mined.After treating S.aureus SA1 with different concentrations of MmEVs,the cell morphology of S.aureus SA1 was observed using transmission electron microscopy.The effects of MmEVs on S.aureus SA1 under low pH(pH value=5)or heat stress(58℃)were investigated.The hydro-phobicity index was explored using the microbial adhesion to hydrocarbons(MATH)assay.Bacte-rial conductivity was measured.The expression levels of biofilm-related genes(SarA,icaB,FnbA,ClfB,CidA,and gyrB)were detected using quantitative real-time PCR(qPCR).The results showed that MIC of MmEVs against the biofilm of S.aureus SA1 was 1 000 mg/L,and the MBIC was 500 mg/L.Under the influence of MmEVs,the internal substances of S.aureus SA1 leaked,the biofilm boundary became blurred,and the cell wall separated.At the MBIC concentration,MmEVs significantly reduced the tolerance of S.aureus SA1 to low pH(P<0.001)and high tem-perature(P<0.001),decreased hydrophobicity(P<0.001),and increased bacterial conductivity(P<0.001).At the MBIC concentration,MmEVs significantly downregulated the gene expression of Sa rA(P<0.001),icaB(P<0.001),FnbA(P<0.001),ClfB(P<0.001),and CidA(P<0.001)in S.aureus SA1,while no significant effect was observed on the expression of the gyrB gene.In summary,MmEVs inhibit the formation of Staphylococcus aureus SA1 biofilms by sup-pressing the gene expression of SarA,icaB,FnbA,ClfB,and CidA within the biofilm.This dis-ruption damages the biofilm's morphological structure,reduces its tolerance to low pH and high temperature,decreases hydrophobicity,and increases bacterial conductivity,thereby ultimately in-hibiting the formation of S.aureus SA1 biofilms.
3.Causal association between immune cells and sepsis: a based on Mendelian randomization method study
Qiushuang YU ; Lingxu LI ; Yina TAO ; Longqiang ZHANG ; Junfeng HU ; Huaxue WANG
Chinese Critical Care Medicine 2024;36(8):821-828
Objective:To investigate the causal association between immune cell and different types of sepsis by using Mendelian randomization (MR) method, and to find the immune cell phenotypes causally associated with sepsis.Methods:Summary data for various circulating immune cell phenotypes were obtained from the GWAS catalog (GCST90001391-GCST90002121). Sepsis data were sourced from the UK Biobank database. Single nucleotide polymorphisms (SNP) were used as instrumental variables. The correlation threshold of P < 5×10 -6 was used to identify the strongly correlated instrumental variables, and the code was used to remove the linkage disequilibrium and the instrumental variables with F-value < 10. Inverse variance weighting (IVW) was used as the main research method to evaluate the stability and reliability of the results, including Cochran's Q test, MR-Egger regression and Leave one out. Reverse MR analysis was performed based on the immunophenotypic results of the removal of horizontal pleiotropy, and the immune cell phenotype with one-way causal association was obtained. Odds ratio ( OR) and 95% confidence interval (95% CI) were used to represent the effect value of the results. Results:CD16 on CD14 -CD16 + monocyte had horizontal pleiotropy in sepsis ( OR = 0.965?4, 95% CI was 0.933?5-0.998?3, P = 0.039?6). There were five immunophenotypes that had reverse causal associations with the types associated with sepsis. After excluding immune cell phenotypes with horizontal pleiotropy and reverse causation, a total of 42 immune cell phenotypes with sepsis, 36 immune cell phenotypes with sepsis (28-day death in critical care), 32 immune cell phenotypes with sepsis (critical care), 44 immune cell phenotypes with sepsis (28-day death), and 30 immune cell phenotypes had potential causal associations with sepsis (under 75 years old). After false discovery rate (FDR) correction, the correlations between BAFF-R on IgD - CD38br and sepsis (28-day death) were negative and strong ( OR = 0.737?8, 95% CI was 0.635?9-0.856?0, P = 6.05×10 -5, PFDR = 0.044?2). Conclusion:A variety of immune cell phenotypes may have a protective effect on sepsis, especially BAFF-R on IgD - CD38br expression is negatively correlated with sepsis (28-day death), which provides a new idea for immune modulation therapy in sepsis.
4.Clinical analysis of different types of neonatal sepsis: a multi-center retrospective study
Yuanqiang YU ; Qingyi DONG ; Suping LI ; Huaxue QI ; Xin TAN ; Hong OUYANG ; Jintao HU ; Wen LI ; Tao WANG ; Yonghui YANG ; Xiaoyun GONG ; Xiaori HE ; Pingyang CHEN
Chinese Journal of Neonatology 2023;38(5):257-261
Objective:To study the clinical characteristics of different types of neonatal sepsis.Methods:From January 2012 to December 2019, neonates with confirmed sepsis from 5 neonatal centers of central-south China were reviewed. The neonates were assigned into early-onset sepsis (EOS) and late-onset sepsis (LOS) group, and the latter was further subgrouped into hospital-acquired LOS (hLOS) group and community-acquired LOS (cLOS) group. The etiological and clinical characteristics were analyzed. SPSS 26.0 was used for statistical analysis.Results:A total of 580 neonates were enrolled, including 286 (49.3%) in the EOS group and 294 (50.7%) in the LOS group. In LOS group, 147 were in hLOS group and 147 were in cLOS group. The gestational age and birth weight of hLOS group were significantly lower than the other two groups [(32.7±3.6) weeks vs. (37.1±3.7) weeks and (37.7±3.0) weeks, (1 810±717) g vs. (2 837±865) g and (3 024±710) g] ( P<0.05). The common pathogens in EOS and cLOS groups were coagulase-negative staphylococci and Escherichia coli, while Klebsiella pneumoniae was common in hLOS group. Carbapenems usage in the hLOS group was significantly higher than the other two groups [62.6% vs. 28.7% and 16.2%] ( P<0.05). Antibiotics duration in the hLOS group was longer than the other two groups [19 (14, 27) d vs. 15 (12, 20) d and 14 (12, 19) d] ( P<0.05). Conclusions:The clinical characteristics of neonatal sepsis vary among different types of infections, and it is necessary to establish appropriate prevention, control, diagnosis and treatment protocols.
5.A case of peripartum cardiomyopathy with recurrent ventricular fibrillation rescued by extracorporeal membrane oxygenation
Shijuan DUN ; Kun LU ; Huaxue WANG
Chinese Journal of Perinatal Medicine 2022;25(12):954-957
Peripartum cardiomyopathy (PPCM) often develops in the early postpartum period, especially within one month after delivery. However, in recent years, some cases of pregnancy-related cardiomyopathy with earlier or slightly later onset showed similar features with PPCM entirely. One patient with PPCM was admitted to the Department of Intensive Care Medicine of the First Affiliated Hospital of Bengbu Medical College on February 2022. After admission, the patient was exceptionally hemodynamically unstable and had repeated ventricular fibrillation. Apart from extracorporeal membrane oxygenation (ECMO) support, comprehensive treatment with cardiotonic agents and drugs to inhibit the "ventricular electric storm" was given. The cardiac structure and function were gradually improved. ECMO was successfully withdrawn on the 7th day after the onset of symptoms, and the patient was discharged after a full recovery. Early diagnosis and active intervention are the keys to improve the prognosis of patients with PPCM and avoid irreversible changes in cardiac structure.
6.Complete ileus due to montmorillonite powder
Adverse Drug Reactions Journal 2022;24(1):42-43
A 50-year-old male patient had diarrhea due to unclean diet. According to the doctor′s advice, the patient received the first dose of montmorillonite powder 6 g orally. The next day, he developed abdominal pain, abdominal distention, and no defecation. Enema was ineffective. Physical examination showed abdominal distension, percussion drum sound, obvious abdominal tenderness, and reduced bowel sound. The patient was diagnosed with complete ileus by abdominal CT. After 5 days of symptomatic treatments such as diet prohibition, gastrointestinal decompression, and enema defecation, the patient returned to normal exhaust and defecation. After excluding other intestinal lesions through abdominal X-ray, enhanced CT, and enteroscopy, it is considered that the patient′s complete ileus was related to montmorillonite powder.
7.Complete ileus due to montmorillonite powder
Adverse Drug Reactions Journal 2022;24(1):42-43
A 50-year-old male patient had diarrhea due to unclean diet. According to the doctor′s advice, the patient received the first dose of montmorillonite powder 6 g orally. The next day, he developed abdominal pain, abdominal distention, and no defecation. Enema was ineffective. Physical examination showed abdominal distension, percussion drum sound, obvious abdominal tenderness, and reduced bowel sound. The patient was diagnosed with complete ileus by abdominal CT. After 5 days of symptomatic treatments such as diet prohibition, gastrointestinal decompression, and enema defecation, the patient returned to normal exhaust and defecation. After excluding other intestinal lesions through abdominal X-ray, enhanced CT, and enteroscopy, it is considered that the patient′s complete ileus was related to montmorillonite powder.
8.Protective effect and mechanism of Angong Niuhuang pill in sepsis-associated brain dysfunction of rats
Ximing DENG ; Qi ZOU ; Shengyong ZHENG ; Huaxue WANG
Chinese Critical Care Medicine 2021;33(8):979-984
Objective:To observe the protective effect of Angong Niuhuang pill on brain function of rats with sepsis, explore its protective mechanism, and provide the experimental basis for clinical application of Angong Niuhuang pill in the treatment of sepsis-associated encephalopathy (SAE).Methods:Thirty male Sprague-Dawley (SD) rats were divided into sham operation group, sepsis model group and Angong Niuhuang pill group according to random number table method, with 10 rats in each group. The sepsis model was established by cecal ligation and puncture (CLP); rats in sham operation group received open and closed abdomen. The rats in the Angong Niuhuang pill group were given Angong Niuhuang pill (0.3 g/kg) by gastric irrigation daily for 3 days before CLP, and the drugs were administrated 12 hours after modeling again. After 24 hours of CLP, the neuroreflex scores were evaluated, white blood cell count (WBC), the levels of serum neuron-specific enolase (NSE) and S100β were detected. Then the brain tissue was harvested. After hematoxylin-eosin (HE) staining, the pathological changes of brain tissue were observed under the light microscope. The mRNA expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in brain tissue were detected by polymerase chain reaction.Results:Compared with the sham operation group, the total score of neuroreflex scores in the sepsis model group and the Angong Niuhuang pill group were significantly reduced (4.43±1.40, 6.57±1.90 vs. 9.40±0.84, both P < 0.05), WBC, serum NSE, S100β were significantly increased [WBC (×10 9/L): 8.07±1.32, 5.84±0.94 vs. 3.60±0.32; NSE (μg/L): 1.04±0.14, 0.61±0.07 vs. 0.16±0.04; S100β (ng/L): 255.624±30.25, 97.72±15.41 vs. 46.88±12.03, all P < 0.05], and the mRNA expressions of IL-6 and TNF-α in brain tissue were significantly increased [IL-6 mRNA (2 -ΔΔCt): 5.668±2.195, 3.605±1.014 vs. 0.997±0.329; TNF-α mRNA (2 -ΔΔCt): 18.996±0.913, 1.746±0.710 vs. 0.674±0.132, all P < 0.05]. Compared with the sepsis model group, the total score of neuroreflex scores in the Angong Niuhuang pill group was significantly increased (6.57±1.90 vs. 4.43±1.40, P < 0.05), WBC, serum NSE, S100β concentration, and the mRNA expressions of IL-6 and TNF-α in the brain were significantly reduced [WBC (×10 9/L): 5.84±0.94 vs. 8.07±1.32, NSE (μg/L): 0.61±0.07 vs. 1.04±0.14, S100β (ng/L): 97.72±15.41 vs. 255.62±30.25, IL-6 mRNA (2 -ΔΔCt): 3.605±1.014 vs. 5.668±2.195, TNF-α mRNA (2 -ΔΔCt): 1.746±0.710 vs. 18.996±0.913, all P < 0.05]. Brain histopathological observation showed that the hippocampal neurons in the sepsis model group were disordered arrangement, a large number of neuronal nuclei were contracted, and the tissue was loose with obvious edema. Compared with the sepsis model group, the Angong Niuhuang pill group had less nuclear shrinkage and tissue edema. Conclusions:The pretreatment of the Angong Niuhuang pill can improve the brain dysfunction of septic rats and reduce the expression of pro-inflammatory cytokines in the brain. It is speculated that the Angong Niuhuang pill can protect the brain function in sepsis by inhibiting the inflammatory reaction in the brain.
9.Clinical efficacy of restrictive fluid management in patients with severe traumatic brain injury.
Shibing ZHAO ; Decai XU ; Rui LI ; Qi ZOU ; Zhenzhen CHEN ; Huaxue WANG ; Xiandi HE
Journal of Southern Medical University 2021;41(1):111-115
OBJECTIVE:
To investigate the effects of restrictive fluid management in patients with severe traumatic brain injury (sTBI).
METHODS:
Between January, 2019 and June, 2020, we randomly assigned 51 postoperative patients (stay in the ICU of no less than 7 days) with sTBI into treatment group (
RESULTS:
The cumulative fluid balance of the two groups were positive on day 1 and negative on days 3 and 7 after ICU admission; at the same time points, the patients in the treatment group had significantly greater negative fluid balance than those in the control group (
CONCLUSIONS
Restrictive fluid management can reduce cerebral edema and improve the prognosis but does not affect the 28-day mortality of patients with sTBI.
Brain Injuries, Traumatic/therapy*
;
Fluid Therapy
;
Humans
;
Prognosis
;
Respiration, Artificial
;
Treatment Outcome
10.Design and application of Checklist for quality control in intensive care unit
Shibing ZHAO ; Qi ZOU ; Chao ZHANG ; Lunjun ZHANG ; Qiang WU ; Ximing DENG ; Huaxue WANG
Chinese Critical Care Medicine 2021;33(4):466-471
Objective:To design a Checklist for quality control in intensive care unit and observe the effect of clinical application.Methods:By consulting guidelines and literature, such as Critical care medicine professional medical quality control index (2015 edition), the quality control Checklist of intensive care unit was designed. It included four parts: quality control data collection, medical record quality verification, special diagnosis and treatment, and hospital infection prevention and control supervision. Every month, a doctor with a senior professional title served as the quality control director, and was responsible for the quality control of the department's medical care, including collecting data of the past 24 hours during the morning handover, discussing and registering special diagnosis and treatment behaviors that would be performed on the day, and coordinating with the nursing team leader, controlling the quality of the whole department throughout the day, such as supervising each medical staff if they had unreasonable behaviors, checking the running and discharge medical records, and inspecting the status of the staff on duty. The data in 2018, 2019 (Checklist implemented) and 2017 (Checklist not implemented) were retrospectively analyzed, including the status of admitted patients, department management information, length of intensive care unit (ICU) stay, and the incidence of three-tube infection [ventilator-associated pneumonia (VAP), catheter-related bloodstream infection (CRBSI), catheter-associated urinary tract infection (CAUTI)], and standardized mortality, etc. Results:From 2017 to 2019, the number of patients admitted was 373, 446, and 480, with annual growth of 19.57% and 7.62% in 2018 and 2019, respectively, and an increase of 28.69% in 2019 compared with 2017. There was no statistically significant difference in the average age and acute physiology and chronic health evaluationⅡ (APACHEⅡ) of patients in the three years. Compared with 2017, the length of ICU stay of patients in 2018 and 2019 were significantly shortened (days: 8.99±6.12, 9.14±7.02 vs. 10.20±7.21), and the incidence of VAP, CRBSI and CAUTI were significantly reduced [VAP (cases/1 000 ventilation days): 12.97±3.60, 9.62±3.14 vs. 17.48±4.89, CRBSI (cases/1 000 catheter days): 3.75±2.19, 3.87±1.87 vs. 6.19±3.13, CAUTI (cases/1 000 catheter days): 3.29±2.18, 3.28±1.87 vs. 5.61±3.18]. The standardized mortality were also significantly reduced [(77.27±7.24)%, (70.61±7.49)% vs. (84.41±9.05)%], the number of non-compliance with hospital infection prevention per month decreased significantly (person times: 54.00±6.30, 41.08±10.76 vs. 72.08±19.68), and the number of special diagnosis and treatment per month increased significantly (person times: 1 056.67±235.27, 1 361.75±278.48 vs. 722.25±145.96), the rate of etiology submission before antimicrobial treatment [(93.21±3.68)%, (96.59±2.49)% vs. (87.86±5.28)%] and deep vein thrombosis (DVT) prevention rate [(91.13±6.36)%, (96.23±2.99)% vs. (85.58±7.68)%] were significantly improved, and all the differences were statistically significant (all P < 0.05). All medical records in the three years were Grade A, but the average scores in 2018 and 2019 were higher than those in 2017 (96.82±2.84, 96.73±2.94 vs. 93.70±3.33, both P < 0.01). Compared with 2018, the incidence of VAP, the rate of etiology submission before antimicrobial treatment, the DVT prevention rate, and the standardized mortality rate in 2019 were further improved, and the number of non-compliance with hospital infection prevention per month decreased and the number of special diagnosis and treatment per month increased, and the differences were statistically significant (all P < 0.05). Conclusion:The application of quality control Checklist in intensive care unit can build an effective quality control system, reduce the incidence of three-tube infection, standardized mortality and length of ICU stay, improve the quality control awareness and execution of medical staff, and promote the improvement of medical quality.

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