Objective:To investigate the effects of Vibrio vulnificus ( Vv) quorum-sensing protein LuxS on the homeostasis of pulmonary innate immune cells in sepsis-induced acute lung injury. Methods:This study constructed luxS knockout and complemented Vv strains. The time required for wild type, luxS knockout, and complemented Vv strains to grow to an absorbance of 600 nm in liquid medium was measured using a spectrophotometer. Iron-overloaded mice were intraperitoneally infected with 1×10 5 CFU of the above three kinds of Vv strains, respectively. Clinical scoring for sepsis-induced dyspnea was used to evaluate the respiratory quality in mice. At 7 h after infection, the pathological changes in lung tissues were observed by HE staining; the bacterial loads in lung tissues were measured; the single-cell suspension of lung tissues was analyzed by flow cytometry. Uniform manifold approximation and projection (UMAP) was used to reduce the dimension of the distribution of CD45 + immune cells in lung tissues of mice in the PBS control group and infection groups with different strains. The frequency and absolute number of innate immune cells in lung tissues were analyzed by multicolor flow cytometry. One-way analysis of variance and t test were used for statistical analysis. Results:There was no significant difference in the growth rate of wild type, luxS knockout, and complemented Vv strains in liquid medium. Compared with the mice infected with the wild type or complemented strain, the mice infected with the luxS knockout strain exhibited overall alleviated respiratory difficulty, decreased inflammatory cell infiltration in lung tissues, and reduced bacterial load in lung tissues ( P<0.05). Besides, there was no significant difference in clinical respiratory scores, inflammatory cell infiltration, or bacterial loads between the mice infected with the complemented strain and wild type strain. UMAP analysis showed that compared with the mice infected with the luxS knockout strain, the mice infected with the wild type or complemented strain showed increased proportions of neutrophils and eosinophils in lung tissues. Results of multicolor flow cytometry analysis further verified that the proportions of neutrophils and eosinophils were significantly lower in the mice infected with the luxS knockout strain than in the mice infected with wild type or complemented strain ( P<0.01, P<0.000 1), while the proportion of alveolar macrophages was significantly higher as compared with that in the mice infected with wild type or complemented strain ( P<0.01). Conclusion:During Vv infection, LuxS may promote acute lung injury by affecting the homeostasis of neutrophils, eosinophils and resident macrophages in lung tissues.

Objective:To investigate the effects of Vibrio vulnificus ( Vv) quorum-sensing protein LuxS on the homeostasis of pulmonary innate immune cells in sepsis-induced acute lung injury. Methods:This study constructed luxS knockout and complemented Vv strains. The time required for wild type, luxS knockout, and complemented Vv strains to grow to an absorbance of 600 nm in liquid medium was measured using a spectrophotometer. Iron-overloaded mice were intraperitoneally infected with 1×10 5 CFU of the above three kinds of Vv strains, respectively. Clinical scoring for sepsis-induced dyspnea was used to evaluate the respiratory quality in mice. At 7 h after infection, the pathological changes in lung tissues were observed by HE staining; the bacterial loads in lung tissues were measured; the single-cell suspension of lung tissues was analyzed by flow cytometry. Uniform manifold approximation and projection (UMAP) was used to reduce the dimension of the distribution of CD45 + immune cells in lung tissues of mice in the PBS control group and infection groups with different strains. The frequency and absolute number of innate immune cells in lung tissues were analyzed by multicolor flow cytometry. One-way analysis of variance and t test were used for statistical analysis. Results:There was no significant difference in the growth rate of wild type, luxS knockout, and complemented Vv strains in liquid medium. Compared with the mice infected with the wild type or complemented strain, the mice infected with the luxS knockout strain exhibited overall alleviated respiratory difficulty, decreased inflammatory cell infiltration in lung tissues, and reduced bacterial load in lung tissues ( P<0.05). Besides, there was no significant difference in clinical respiratory scores, inflammatory cell infiltration, or bacterial loads between the mice infected with the complemented strain and wild type strain. UMAP analysis showed that compared with the mice infected with the luxS knockout strain, the mice infected with the wild type or complemented strain showed increased proportions of neutrophils and eosinophils in lung tissues. Results of multicolor flow cytometry analysis further verified that the proportions of neutrophils and eosinophils were significantly lower in the mice infected with the luxS knockout strain than in the mice infected with wild type or complemented strain ( P<0.01, P<0.000 1), while the proportion of alveolar macrophages was significantly higher as compared with that in the mice infected with wild type or complemented strain ( P<0.01). Conclusion:During Vv infection, LuxS may promote acute lung injury by affecting the homeostasis of neutrophils, eosinophils and resident macrophages in lung tissues.

BACKGROUND:The traditional method for treating infected bone defects is thorough debridement,filling with antibiotic-impregnated bone cement,and then autologous bone transplantation.However,there are problems such as insufficient antibiotic concentration and drug resistance,and limited bone mass.Therefore,it is of great clinical significance to seek dual-functional biomaterials that can locally release antibiotics at the site of infection and promote bone repair.OBJECTIVE:To design an injectable hydrogel composed of hyaluronic acid and oxidized dextran as a local delivery system for vancomycin to treat infection and promote bone regeneration.METHODS:(1)Vancomycin-loaded hyaluronic acid/oxidized dextran hydrogels were prepared to characterize the morphology,mechanical properties,and in vitro drug release of the hydrogels.(2)Hyaluronic acid/oxidized dextran hydrogels and vancomycin-loaded hyaluronic acid/oxidized dextran hydrogels were co-cultured with rabbit bone marrow mesenchymal stem cells.Cell activity and proliferation were detected by live-dead staining and CCK-8 assay.After osteogenic induction,alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-qPCR detection(osteocalcin and bone morphogenetic protein 2 mRNA expression)were performed to evaluate the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.(3)The above two hydrogels were co-cultured with Escherichia coli(or Staphylococcus aureus)to detect the antibacterial ability of the hydrogels.(4)Thirty rabbits were selected to establish a 1.5 cm infected bone defect model in the middle of the left radius.Two weeks after modeling,they were randomly divided into three intervention groups:the blank group(n=10)did not receive any treatment;the control group(n=10)was injected with hyaluronic acid/oxidized dextran hydrogel at the bone defect site;the experimental group(n=10)was injected with vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel at the bone defect site.12 weeks after injection,the samples were collected for Micro-CT scanning,tissue morphology observation,RT-qPCR detection(tumor necrosis factor α,interleukin 6 mRNA expression),and immunohistochemical staining.RESULTS AND CONCLUSION:(1)Vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel had a good porous structure with a pore size between 100-300 μm,and had good mechanical properties and in vitro drug sustained release performance.(2)Live-dead staining and CCK-8 assay results confirmed that vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel had good biocompatibility.Alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-qPCR test results showed that both hydrogels could promote osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.(3)Compared with hyaluronic acid/oxidized dextran hydrogel,vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel could significantly inhibit the growth of Escherichia coli and Staphylococcus aureus.(4)Micro-CT scanning results showed that the bone volume fraction and bone density of new bone in the bone defect area of the experimental group were higher than those of the blank group and the control group(P＜0.05).The bone tissue morphology observation results showed that the experimental group had better repair effect of bone defects compared with the blank group and control group.The expression of tumor necrosis factor α and interleukin 6 mRNA in the bone defect site in the experimental group was lower than that in the blank group and control group(P＜0.05).Immunohistochemical staining showed that the protein expressions of osteocalcin and RUNX2 at the site of bone defects in the experimental group and control group were higher than those in the blank group(P＜0.05).These findings indicate that vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel can effectively promote bone regeneration under infection.

Objective:To investigate microstructural alterations in the optic chiasm and optic radiations of multiple sclerosis (MS) and neuromyelitis optica spectrum disorders (NMOSD) based on diffusion kurtosis imaging (DKI).Methods:This study was a cross-sectional study. Retrospective analyses were conducted on the clinical and imaging data of 63 patients with relapsing-remitting MS (RRMS) and 62 patients with NMOSD diagnosed at First Affiliated Hospital of Chongqing Medical University from January 2019 to December 2023. According to the occurrence of optic neuritis (ON), they were categorized into ON-positive MS (ON+MS) group (40 cases), ON-negative MS (ON-MS) group (23 cases), ON-positive NMOSD (ON+NMOSD) group (40 cases) and ON-negative NMOSD (ON-NMOSD) group (22 cases). In addition, 40 healthy controls were enrolled during the same period. DKI data of all subjects were collected, and DKI post-processing was performed to obtain fractional anisotropy (FA), mean kurtosis (MK), axial kurtosis (AK), and radial kurtosis (RK) values of the optic chiasm and bilateral optic radiations. The scores of the mini-mental state examination (MMSE), montreal cognitive assessment (MoCA), and expanded disability status scale (EDSS) were obtained. The Kruskal-Wallis test was used to analyze the differences in DKI parameters of the optic chiasm and bilateral optic radiation among the 5 groups, and the Holm-Bonferroni method was employed for multiple comparison correction in pairwise comparisons.Results:There were statistically significant overall differences in the DKI parameters of the optic chiasm and bilateral optic radiations among healthy control group, ON+MS group, ON-MS group, ON+NMOSD group, and ON-NMOSD group (all P0.05). The FA value of the optic chiasm in ON+NMOSD group was significantly lower than that of healthy control group and ON-MS group, as well as ON-NMOSD group ( P0.05). The FA value of the left optic radiation in ON+NMOSD group was lower than that in healthy control group and the ON-MS group. The RK value of the optic chiasm in ON+MS group was lower than that in the healthy control group and ON-NMOSD group ( P0.05). The MK and RK values of the left optic radiation in ON-MS group were significantly lower than those in the ON+NMOSD group and ON-NMOSD group ( P0.05). Conclusions:NMOSD and RRMS patients demonstrate varying degrees of microstructural damage in the optic chiasm and optic radiations. Differences of DKI parameters suggest different pathological mechanisms of visual pathway damage between NMOSD and MS, which may be helpful for early detection of occult visual pathway lesions.

Objective:To investigate the efficacy and safety of endovascular therapy (EVT) guided by noncontrast CT (NCCT) in patients with large vessel occlusive stroke within a late window (6-24 hours after onset).Methods:Consecutive patients with acute large vessel occlusive stroke within a late window admitted to the General Hospital of Wanbei Coal-electricity Group Co., Ltd from July 1, 2023 to September 30, 2024 were included retrospectively. All patients completed CT angiography or magnetic resonance angiography to confirm the presence of anterior circulation large vessel occlusion and rule out intracranial hemorrhage (ICH). The Alberta Stroke Program Early CT Score (ASPECTS) ≥4 as assessed by NCCT images was used as the EVT screening criteria. The main outcome measure was the functional outcome assessed by the modified Rankin Scale at 90 days after onset, 0-2 was defined as good outcome. The secondary outcome measures were symptomatic ICH (sICH) and death within 90 days. Multivariate logistic regression analysis was used to determine the independent influencing factors of functional outcome. Results:A total of 74 patients with large vessel occlusive stroke within the late window were enrolled, including 42 males (56.8%), aged 68.69±11.62 years (range, 47-89 years), with a baseline National Institutes of Health Stroke Scale score 11.20±5.12 and a median baseline ASPECTS 7 (interquartile range, 6-8). Ten (13.5%) and 7 patients (9.5%) respectively experienced any ICH and sICH, and 7 (9.5%) died within 90 days after onset. Twenty-seven patients (36.5%) received EVT, 47 (63.5%) only received conventional drug treatment; 20 (27.0%) had good outcome, and 54 (73.0%) had poor outcome. Univariate analysis showed that the good outcome rate in the EVT group at 90 days was significantly higher than that in the non-EVT group (40.7% vs. 19.1%; χ2=4.054, P=0.044), and there was no significant difference in the proportion of patients in sICH (11.1% vs. 8.5%; χ2=0.701, P=0.505) and who died within 90 days (7.4% vs. 10.6%; χ2=1.000, P=0.495) compared to the non-EVT group. The proportion of patients receiving EVT in the good outcome group was significantly higher than that in the poor outcome group (60.0% vs. 27.8%; χ2=6.539, P=0.011). Multivariate logistic regression analysis showed that EVT was an independent influencing factor of good outcome (odds ratio 0.440, 95% confidence interval 0.144-0.987; P=0.041). Conclusion:Compared with the conventional drug treatment alone, EVT guided by NCCT evaluation can achieve better outcome in patients with large vessel occlusion stroke within late window, and does not increase the risk of sICH and death within 90 days.

BACKGROUND:The traditional method for treating infected bone defects is thorough debridement,filling with antibiotic-impregnated bone cement,and then autologous bone transplantation.However,there are problems such as insufficient antibiotic concentration and drug resistance,and limited bone mass.Therefore,it is of great clinical significance to seek dual-functional biomaterials that can locally release antibiotics at the site of infection and promote bone repair.OBJECTIVE:To design an injectable hydrogel composed of hyaluronic acid and oxidized dextran as a local delivery system for vancomycin to treat infection and promote bone regeneration.METHODS:(1)Vancomycin-loaded hyaluronic acid/oxidized dextran hydrogels were prepared to characterize the morphology,mechanical properties,and in vitro drug release of the hydrogels.(2)Hyaluronic acid/oxidized dextran hydrogels and vancomycin-loaded hyaluronic acid/oxidized dextran hydrogels were co-cultured with rabbit bone marrow mesenchymal stem cells.Cell activity and proliferation were detected by live-dead staining and CCK-8 assay.After osteogenic induction,alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-qPCR detection(osteocalcin and bone morphogenetic protein 2 mRNA expression)were performed to evaluate the osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.(3)The above two hydrogels were co-cultured with Escherichia coli(or Staphylococcus aureus)to detect the antibacterial ability of the hydrogels.(4)Thirty rabbits were selected to establish a 1.5 cm infected bone defect model in the middle of the left radius.Two weeks after modeling,they were randomly divided into three intervention groups:the blank group(n=10)did not receive any treatment;the control group(n=10)was injected with hyaluronic acid/oxidized dextran hydrogel at the bone defect site;the experimental group(n=10)was injected with vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel at the bone defect site.12 weeks after injection,the samples were collected for Micro-CT scanning,tissue morphology observation,RT-qPCR detection(tumor necrosis factor α,interleukin 6 mRNA expression),and immunohistochemical staining.RESULTS AND CONCLUSION:(1)Vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel had a good porous structure with a pore size between 100-300 μm,and had good mechanical properties and in vitro drug sustained release performance.(2)Live-dead staining and CCK-8 assay results confirmed that vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel had good biocompatibility.Alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-qPCR test results showed that both hydrogels could promote osteogenic differentiation of rabbit bone marrow mesenchymal stem cells.(3)Compared with hyaluronic acid/oxidized dextran hydrogel,vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel could significantly inhibit the growth of Escherichia coli and Staphylococcus aureus.(4)Micro-CT scanning results showed that the bone volume fraction and bone density of new bone in the bone defect area of the experimental group were higher than those of the blank group and the control group(P＜0.05).The bone tissue morphology observation results showed that the experimental group had better repair effect of bone defects compared with the blank group and control group.The expression of tumor necrosis factor α and interleukin 6 mRNA in the bone defect site in the experimental group was lower than that in the blank group and control group(P＜0.05).Immunohistochemical staining showed that the protein expressions of osteocalcin and RUNX2 at the site of bone defects in the experimental group and control group were higher than those in the blank group(P＜0.05).These findings indicate that vancomycin-loaded hyaluronic acid/oxidized dextran hydrogel can effectively promote bone regeneration under infection.

Objective:To investigate microstructural alterations in the optic chiasm and optic radiations of multiple sclerosis (MS) and neuromyelitis optica spectrum disorders (NMOSD) based on diffusion kurtosis imaging (DKI).Methods:This study was a cross-sectional study. Retrospective analyses were conducted on the clinical and imaging data of 63 patients with relapsing-remitting MS (RRMS) and 62 patients with NMOSD diagnosed at First Affiliated Hospital of Chongqing Medical University from January 2019 to December 2023. According to the occurrence of optic neuritis (ON), they were categorized into ON-positive MS (ON+MS) group (40 cases), ON-negative MS (ON-MS) group (23 cases), ON-positive NMOSD (ON+NMOSD) group (40 cases) and ON-negative NMOSD (ON-NMOSD) group (22 cases). In addition, 40 healthy controls were enrolled during the same period. DKI data of all subjects were collected, and DKI post-processing was performed to obtain fractional anisotropy (FA), mean kurtosis (MK), axial kurtosis (AK), and radial kurtosis (RK) values of the optic chiasm and bilateral optic radiations. The scores of the mini-mental state examination (MMSE), montreal cognitive assessment (MoCA), and expanded disability status scale (EDSS) were obtained. The Kruskal-Wallis test was used to analyze the differences in DKI parameters of the optic chiasm and bilateral optic radiation among the 5 groups, and the Holm-Bonferroni method was employed for multiple comparison correction in pairwise comparisons.Results:There were statistically significant overall differences in the DKI parameters of the optic chiasm and bilateral optic radiations among healthy control group, ON+MS group, ON-MS group, ON+NMOSD group, and ON-NMOSD group (all P0.05). The FA value of the optic chiasm in ON+NMOSD group was significantly lower than that of healthy control group and ON-MS group, as well as ON-NMOSD group ( P0.05). The FA value of the left optic radiation in ON+NMOSD group was lower than that in healthy control group and the ON-MS group. The RK value of the optic chiasm in ON+MS group was lower than that in the healthy control group and ON-NMOSD group ( P0.05). The MK and RK values of the left optic radiation in ON-MS group were significantly lower than those in the ON+NMOSD group and ON-NMOSD group ( P0.05). Conclusions:NMOSD and RRMS patients demonstrate varying degrees of microstructural damage in the optic chiasm and optic radiations. Differences of DKI parameters suggest different pathological mechanisms of visual pathway damage between NMOSD and MS, which may be helpful for early detection of occult visual pathway lesions.

Objective To investigate the predictive value of the inflammation scoring system based on the blood routine for complicated acute pancreatitis(CAP).Methods The clinical data of 630 patients with a-cute pancreatitis(AP)receiving the treatment in the People's Hospital of Chongqing Liang Jiang New Area from January 2021 to January 2024 were retrospectively analyzed.The patients were divided into the mild a-cute pancreatitis(MAP)group and CAP group according to the disease severity degree.The propensity score matching was used to balance the baseline characteristics of the cases in the two groups,the differences be-tween the inflammatory scoring system based on the blood routine[neutrophile granulocyte/lymphocyte ratio(NLR),derived neutrophile granulocyte/lymphocyte ratio(dNLR),platelet/lymphocyte ratio(PLR),system-ic immune inflammatory index(SII)and systemic inflammatory response index(SIRI)]and the clinical out-come indexes within 24 h of admission were analyzed,and the logistic regression was used to analyze the risk factors of CAP.Results A total of 404 patients were included in the study by the propensity score matching,each 202 cases in the two groups.In the CAP group,there were 35 cases(16.7％)receiving intensive care unit(ICU)treatment and 4 cases(1.9％)of death.Compared with the MAP group,WBC,neutrophile granulo-cyte,CRP,BUN,amylase and lipase levels and NLR,dNLR,SII and SIRI scores in the CAP group were high-er,serum calcium and albumin levels were lower,and the differences were statistically significant(P＜0.05).The stepwise logistics regression analysis showed that CRP,albumin,lipase,serum calcium and SIRI were the independent influencing factors of CAP occurrence(P＜0.05).Conclusion The inflammation scoring system based on the blood routine could predict the CAP occurrence.

Objective To summarize our experience and the early and midterm outcomes of stented elephant trunk procedure for right-sided aortic arch (RAA) with Kommerell's diverticulum (KD). Methods From April 2013 to July 2020, patients with RAA and KD who underwent stented elephant trunk procedure at our center were collected. Surgery was performed under moderate hypothermic circulatory arrest combined with selective antegrade cerebral perfusion via median sternotomy. Results A total of 8 patients were included, including 7 males and 1 female with a mean age of 51.88±9.61 years. All patients had an aneurysmal KD and aberrant left subclavian artery. Preoperative comorbidities included acute Stanford type B aortic dissection in 1 patient, aortic arch pseudoaneurysm in 1 patient, acute type B intramural hematoma in 2 patients, and coronary artery disease in 1 patient. Concomitant procedures included reconstruction of the left subclavian artery in all patients and coronary artery bypass grafting in 1 patient. The mean time of operation, cardiopulmonary bypass, aortic cross-clamping, and selective cerebral perfusion was 6.25±1.16 h, 157.75±40.07 min, 77.75±33.10 min, and 28.50±5.55 min, respectively. No intraoperative death occurred. There was 1 in-hospital death. Follow-up was completed in all patients with a mean period of 3.58±2.08 years. No late death occurred. A persistent anastomotic leak of the proximal arch was detected in 1 patient, but reintervention was not performed because neither aortic dilatation nor symptoms of tracheal and esophageal compression were observed during the follow-up. The remaining 6 patients showed positive aortic remodeling with complete thrombosis of the aneurysmal KD, and neither aortic event nor tracheal and esophageal compression occurred. Conclusion Stented elephant trunk procedure is a safe and feasible technique for selected patients with RAA and KD, which can achieve favorable early and midterm outcomes.

Objective To evaluate the value of dual-source CT angiography for evaluating the degree of coronary stenosis. Methods A total of 110 patients with a high likelihood of coronary stenosis identified by dual-source CT angiography or conventional coronary angiography were enrolled. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of dual-source CT angiography for diagnosis of coronary stenosis were evaluated with conventional coronary angiography as a gold standard. The agreement between dual-source CT angiography and conventional coronary angiography for evaluation of coronary stenosis was evaluated using Kappa statistic. Results A total of 1 401 coronary artery segments from 110 patients were displayed on conventional coronary angiography, while 1 382 segments were successfully visualized in dual-source CT angiography (98.64%). The sensitivity, specificity, positive predictive value and negative predictive value of dual-source CT angiography were 97.9%, 97.3%, 90.4% and 99.4% for diagnosis of coronary stenosis, and there was high consistence between dual-source CT angiography and conventional coronary angiography for grading coronary stenosis (Kappa statistic = 0.87, U = 58.36, P < 0.01). In addition, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of dual-source CT angiography were 94.7%, 96.8%, 83.7%, 99.0% and 96.5% for grading stenosis of coronary artery segments. Conclusion Dual-source CT angiography is accurate and reliable for diagnosis of coronary stenosis, which may be a non-invasive tool for assessment of coronary stenosis.