Although cancer immunotherapy has made great strides in the clinic, it is still hindered by the tumor immunosuppressive microenvironment (TIME). The stimulator of interferon genes (STING) pathway which can modulate TIME effectively has emerged as a promising therapeutic recently. However, the delivery of most STING agonists, specifically cyclic dinucleotides (CDNs), is performed intratumorally due to their insufficient pharmacological properties, such as weak permeability across cell membranes and vulnerability to nuclease degradation. To expand the clinical applicability of CDNs, a novel pH-sensitive polycationic polymer-modified lipid nanoparticle (LNP-B) system was developed for intravenous delivery of CDNs. LNP-B significantly extended the circulation of CDNs and enhanced the accumulation of CDNs within the tumor, spleen, and tumor-draining lymph nodes compared with free CDNs thereby triggering the STING pathway of dendritic cells and repolarizing pro-tumor macrophages. These events subsequently gave rise to potent anti-tumor immune reactions and substantial inhibition of tumors in CT26 colon cancer-bearing mouse models. In addition, due to the acid-sensitive property of the polycationic polymer, the delivery system of LNP-B was more biocompatible and safer compared with lipid nanoparticles formulated with an indissociable cationic DOTAP (LNP-D). These findings suggest that LNP-B has great potential in the intravenous delivery of CDNs for tumor immunotherapy.

OBJECTIVE: To explore the timing of weaning extracorporeal membrane oxygenation (ECMO) and analyze the risk factors that affect survival outcomes before weaning. METHODS: A retrospective case-control study was conducted. Patients who received ECMO treatment and were weaned according to physicians' orders at the Second Xiangya Hospital of Central South University from January 2020 to June 2024 were enrolled as the study subjects. The general information, underlying diseases, indications and processes of ECMO, vital signs and arterial blood gas analysis 1 hour before weaning test, and biochemical indicators 24 hours before weaning test were collected through the hospital electronic medical record system. The primary outcome measure was the hospital mortality. The variables with P < 0.1 in univariate analysis and correlation analysis were included into binary Logistic regression analysis to identify risk factors. A nomogram model was constructed to predict the risk of weaning death in patients with ECMO, and receiver operator characteristic curve (ROC curve) and calibration curve were drawn to evaluate the model. Decision curve analysis (DCA) was used to evaluate the clinical net benefit rate of the model. RESULTS: A total of 32 ECMO patients were included, among whom 10 received veno-arterial ECMO (VA-ECMO) and 22 received veno-venous ECMO (VV-ECMO). During the hospitalization period, 23 patients survived, while 9 died. The time from mechanical ventilation to ECMO activation in the death group was significantly longer than that in the survival group, and the time from ECMO cessation to discharge was significantly shorter than that in the survival group. The levels of diastolic blood pressure (DBP) and albumin (Alb) before weaning were significantly lower than those in the survival group, and the level of procalcitonin (PCT) was significantly higher than that in the survival group (all P < 0.05). Spearman correlation analysis showed that DBP, PCT, Alb, and thrombin time (TT) were correlated with the weaning outcomes of ECMO patients (r values were -0.450, 0.373, -0.376, -0.346, all P < 0.1). Binary Logistic regression analysis showed that the final indicators entering the regression equation included DBP [odds ratio (OR) = 0.864, 95% confidence interval (95%CI) was 0.756-0.982], PCT (OR = 1.157, 95%CI was 0.679-1.973), and TT (OR = 0.852, 95%CI was 0.693-1.049), and a nomogram model was constructed to predict the weaning outcomes of ECMO patients. ROC curve analysis showed that the area under the curve (AUC) of the nomogram model for predicting the weaning outcome of ECMO patients was 0.831, with a sensitivity of 77.8% and a specificity of 65.2%. Its predictive value was better than that of single indicators DBP, PCT, and TT (AUC of 0.787, 0.739, and 0.722, respectively). The calibration curve showed that the prediction probability of the model was in good consistency with the actual observed results, the Hosmer-Lemeshow goodness of fit test showed that, χ ² = 8.3521, P = 0.400, indicating that the model fits well. DCA showed that across risk threshold of 0-0.8, the net benefit rate was greater than 0, which was significantly better than that of single indicator. CONCLUSIONS The nomogram model constructed with DBP, PCT, and TT has certain predictive value for the weaning outcomes of ECMO patients and can be used as a screening indicator for ECMO weaning timing.
Humans ; Extracorporeal Membrane Oxygenation ; Retrospective Studies ; Risk Factors ; Case-Control Studies ; Hospital Mortality ; Male ; Female ; Nomograms ; Logistic Models ; ROC Curve ; Middle Aged ; Adult ; Ventilator Weaning ; Time Factors

OBJECTIVE: To identify the risk factors related to the timing of patients receiving extracorporeal membrane oxygenation (ECMO) initiation and construct a risk prediction model for ECMO initiation timing. METHODS: Patients who received ECMO admitted to the Second Xiangya Hospital of Central South University from January 2020 to January 2024 were retrospectively collected. The case data mainly included physiological and biochemical indicators 1 hour before ECMO initiation. According to the outcome of the patients, they were divided into survival group and death group. Univariate and multivariate Logistic regression analysis were used to analyze the predictors of mortality risk in patients with ECMO, and a nomogram prediction model was constructed. The discrimination, calibration accuracy, and goodness of the model were evaluated by the receiver operator characteristic curve (ROC curve), calibration curve, and the Hosmer-Lemeshow test, respectively. Decision curve analysis (DCA) evaluated the clinical net benefit rate of the model. RESULTS: A total of 81 ECMO patients were included, including 59 males and 22 females; age range from 16 to 61 years old, with a median age of 56.0 (39.5, 61.5) years old; 20 patients received veno-arterial (V-A) ECMO, and 61 patients received veno-venous (V-V) ECMO; 23 patients ultimately survived and 58 patients died. Univariate analysis showed that age, blood urea nitrogen, serum creatinine, D-dimer, arterial blood carbon dioxide partial pressure, and prothrombin time of the death group were all higher than those of the survival group, while albumin was slightly lower than that of the survival group. There was a statistically significant difference in the direct cause of ECMO initiation between the two groups. Multivariate Logistic regression analysis showed that age [odds ratio (OR) = 1.069, 95% confidence interval (95%CI) was 1.015-1.125, P = 0.012], direct cause of ECMO initiation [with heart failure as the reference, return of spontaneous circulation (ROSC) after cardiopulmonary support (OR = 30.672, 95%CI was 1.265-743.638, P = 0.035), novel coronavirus infection (OR = 8.666, 95%CI was 0.818-91.761, P = 0.073), other severe pneumonia (OR = 4.997, 95%CI was 0.558-44.765, P = 0.150)], pre-ECMO serum creatinine (OR = 1.008, 95%CI was 1.000-1.016, P = 0.044), prothrombin time (OR = 1.078, 95%CI was 0.948-1.226, P = 0.252), and D-dimer (OR = 1.135, 95%CI was 1.047-1.231, P = 0.002) were entered into the final regression equation. A nomogram prediction model was developed based on these five factors. The area under the ROC curve (AUC) of the model was 0.889 (95%CI was 0.819-0.959), higher than the AUC of the sequential organ failure assessment (SOFA; AUC = 0.604, 95%CI was 0.467-0.742). The calibration curve showed good consistency between the model predictions and the observed results. The Hosmer-Lemeshow goodness-of-fit test showed that χ ² = 4.668, P = 0.792. DCA analysis showed that when the risk threshold was 0-0.8, the net benefit rate was greater than 0, which was significantly better than that of SOFA score. CONCLUSIONS The risk prediction model for the timing of ECMO initiation, constructed using five factors (age, direct cause of ECMO initiation, thrombin time, serum creatinine, and D-dimer), demonstrated good discrimination and calibration. It can serve as a pre-initiation assessment tool to identify and predict post-initiation mortality risk in ECMO patients.
Humans ; Extracorporeal Membrane Oxygenation ; Middle Aged ; Male ; Female ; Retrospective Studies ; Adult ; Risk Factors ; Adolescent ; Young Adult ; Logistic Models ; Nomograms ; ROC Curve ; Time Factors ; Risk Assessment

Objective:The sensory processing measure-preschool scale(SPM-P)was transformed into Chinese,and its re-liability and validity were tested in preschool children. Method:According to Brislin's translation model,the SPM-P source scale was translated into Chinese and the Chinese version of SPM-P scale was formed.From September 2021 to December 2021,395 preschool children were investigated by cluster stratified random sampling method to test the reliability and validity of the scale. Result:The Chinese version of the SPM-P scale includes 8 dimensions of social participation,vision,hear-ing,touch,body awareness,balance and movement,planning and conception,and overall sensory system,with a total of 75 items.Cronbach alpha coefficient for the overall scale was 0.899,the split half reliability coefficient was 0.700;the test-retest reliability coefficient was 0.899.The item content validity(I-CVI)value was 0.920,the average content validity(S-CVI/Ave)value was 0.984,and the content validity was good;the results of confirmatory factor analysis showed that the construct validity was good;The results of confirma-tory factor analysis showed that the construct validity was good(x2/df=2.41,CFI=0.992,TLI=0.960,RMSEA=0.060,SRMR=0.046);The Chinese version of SPM-P scale was negatively correlated with the corresponding evaluation dimensions of the screening questionnaire for children's sensory integration disorder,and the correla-tion coefficient was between-0.585 and-0.399,with good criterion validity. Conclusion:The Chinese version of SPM-P scale has good reliability and validity and can be used for stan-dardized evaluation of sensory integration ability of preschool children in China.

Objective:To investigate the risk factors for aneurysm rupture and post-intervention complications in intracranial arterial stenosis patients with intracranial aneurysms.Methods:A retrospective analysis was performed; 238 intracranial arterial stenosis patients with intracranial aneurysms (306 intracranial aneurysms) admitted to Cerebrovascular Disease Department, Neurosurgery Center, Zhujiang Hospital, Southern Medical University from January 2018 to August 2022 were chosen. Ruptured group and unruptured group were divided according to the rupture of intracranial aneurysms. Additionally, 139 patients who underwent interventional therapy and had complete follow-up data were divided into 2 groups according to occurrence of post-intervention complications. Univariate and multivariate Logistic regression analyses were used to identify the risk factors for aneurysm rupture and post-intervention complications.Results:(1) Of 238 patients, 269 unruptured aneurysms and 37 ruptured aneurysms were noted. Univariate regression analysis showed that significant difference was noted between the ruptured group and unruptured group in female ratio, aneurysm distribution, proportion of irregular shaped aneurysms, percentages of patients with increased white blood cell count, neutrophil count, total cholesterol and D-2 polymer, and percentage of patients with decreased blood lymphocyte count ( P<0.05). Multivariate Logistic regression analysis showed that irregular shaped aneurysms ( OR=12.393, 95% CI: 4.114-37.332, P<0.001), elevated neutrophil count ( OR=18.753, 95% CI: 6.555-53.648, P<0.001), and increased D-2 polymer ( OR=4.410, 95% CI: 1.758-11.065, P=0.002) were independent risk factors for aneurysm rupture in intracranial arterial stenosis patients with intracranial aneurysms. (2) Of the 139 patients, 57 had complications and 82 had no complications. Univariate regression analysis showed that the proportion of patients with hypertension history, distribution of arterial stenosis, and proportion of patients with elevated blood D-2 polymer were significantly different between patients with and without complications ( P<0.05); while multivariate Logistic regression analysis did not identify these 3 indexes as independent risk factors for post-intervention complications ( P>0.05). Conclusion:Patients with irregular shaped aneurysms, elevated blood neutrophil count and D-2 polymer trend to have aneurysm rupture; hypertension history, arterial stenosis, and elevated D-2 polymer have impact on postoperative complications in intracranial arterial stenosis patients with intracranial aneurysms.

Objective:This study aimed at investigating the efficacy and safety of eltrombopag in the treatment of adult primary immune thrombocytopenia (ITP) and evaluated the factors influencing its efficacy and side effects.Methods:A total of 198 patients with adult ITP who were admitted to Tianjin Medical University General Hospital between January 2018 and March 2022 were retrospectively analyzed. The efficacy of each starting dose of eltrombopag was evaluated, and adverse events were analyzed. The factors influencing efficacy were investigated, including sex, age, adult ITP type, platelet antibodies, and combined drug treatments.Results:Of the 198 patients, 70 males and 128 females with a median age of 45 years (18-88 years) were included; 130 (65.7%) had newly diagnosed adult ITP, 25 (12.6%) had persistent adult ITP, and 43 (21.7%) had chronic adult ITP. The bleeding event scores at baseline were assessed; 84.3% had scores of<4 and 15.7% had scores of ≥4. The eltrombopag response rate (initial response) at 6 weeks was 78.8% (complete response [CR]: 49.0%; CR1: 14.6%; CR2: 15.2%). The median response time to eltrombopag was 7 (7, 14) days. The initial response rates to 25, 50, and 75 mg eltrombopag were 74.1%, 85.9%, and 60.0%, respectively ( P=0.031). The initial response rate to the 50 mg dose was significantly higher than that of the 25-mg and 75-mg doses. Two patients received 100 mg as the starting dose, and their initial response was 0. Regarding dose adjustment, 70.7% of the patients remained on the starting dose, 8.6% underwent dose adjustment to 50 mg, and 6.1% underwent dose adjustment to 75 mg. Another two patients underwent dose adjustment to 100 mg. After dose adjustment, the persistent response rates were 83.6%, 85.3%, and 85.7% for the 25-, 50-, and 75-mg doses, respectively, with no significant difference. After dose adjustment, the sustained efficacy rate for the 100-mg dose (4 patients) was 100.0%. After 6 weeks of treatment with eltrombopag, the overall bleeding score of patients with ITP decreased. The number of patients with a score of ≥4 decreased to 0, the number of patients with a score of<4 decreased, and there was no significant change in the number of patients with a score of 1-2. The most common adverse event associated with eltrombopag was impaired liver function (7.7%). No thrombosis events or other adverse events were observed. ITP type and number of megakaryocytes significantly affected the initial response to eltrombopag. The initial response rates to eltrombopag for newly diagnosed adult ITP, persistent adult ITP, and chronic adult ITP were 85.3%, 56.0%, and 76.2%, respectively ( P=0.003). For megakaryocytes, the initial response rates were 61.8%, 87.1%, and 84.3% ( P=0.009) for the decreased, normal, and increased megakaryocyte groups, respectively. Conclusion:Eltrombopag, as a second-line or higher treatment for adult ITP, has a rapid onset of action and good safety. The initial response rate is significantly higher with a dose of 50 mg than with a dose of 25 mg. Patients with newly diagnosed ITP and those with normal or increased megakaryocyte numbers have a higher initial response rate to eltrombopag.

In this study, a new Bacillus velezensis strain Bv-303 was identified and its biocontrol effect against rice bacterial-blight (BB) disease caused by Xanthomonas oryzae pv. oryzae (Xoo) was investigated. Cell-free supernatant (CFS) of strain Bv-303 under different growth conditions were prepared to test the antagonistic activity and stability against Xoo by the Oxford-cup method in vitro. The antibacterial effect of strain Bv-303 to BB disease in rice were further analyzed in vivo by spraying the cell-culture broth (CCB), CFS and cell-suspension water (CSW), respectively, on the rice leaves inoculated with Xoo. Additionally, rice seeds germination rate and seedling growth under the strain Bv-303 CCB treatment were tested. The results showed that the strain Bv-303 CFS significantly inhibited Xoo growth by 85.7%‒88.0% in vitro, which was also stable under extreme environment conditions such as heat, acid, alkali and ultraviolet light. As tested in vivo, spraying the CCB, CFS or CSW of strain Bv-303 on the Xoo-infected leaves enhanced rice plant resistance to BB disease, with CCB showing the highest increase (62.7%) in disease-resistance. Notably, CCB does not have negative effects on rice seed germination and seedling growth. Therefore, strain Bv-303 has great potential for biocontrol of the rice BB disease.
Oryza ; Fatigue Syndrome, Chronic ; Bacillus ; Xanthomonas ; Plant Diseases/microbiology*

Objective:To investigate the effect of neutrophils on cell pyroptosis and its mechanisms in mice with early brain injury (EBI) following subarachnoid hemorrhage (SAH).Methods:Seventy six male C57BL/6J mice were randomly divided into sham-operated group, SAH group, SAH+vehicle group, and SAH+anti-ly6G group ( n=19). SAH models in the latter 3 groups were established by modified endovascular perforation. Mice in the SAH+vehicle group and SAH+anti-ly6G group received intravenous injection of equal normal saline or anti-ly6G antibody (4 mg/kg) 24 h before SAH. At 24 h after SAH, immunofluorescent staining was used to detect the locations/expressions of neutrophils, S100 calcium binding protein A8 (S100A8) and gasdermin D (GSDMD); FJC staining was performed to assess the neuronal injury; modified Garcia test and rotarod test were used to evaluate the neurological functions, and brain water content test was applied to evaluate the brain edema; Western blotting was used to detect the expressions of S100A8, Toll-like receptor 4 (TLR4), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), cleaved cysteinyl aspartate specific proteinase-1 (cleaved-caspase1), and cleaved N-terminal gasdermin D (GSDMD-N). Results:(1) Compared with those in the sham-operated group, neutrophil infiltration at the damaged cortex with highly expressed S100A8 in neutrophils was observed in the SAH group, and increased GSDMD expression at the damaged cortex and GSDMD co-localization in astrocytes, microglia and neurons were observed in the SAH group. (2) Compared with the sham-operated group, the SAH group and SAH+vehicle group had significantly increased numbers of infiltrated neutrophils and FJC-positive neurons, significantly decreased falling latency in the modified Garcia score and rotarod test, significantly increased brain water content, and significantly elevated expressions of S100A8, TLR4, NLRP3, cleaved-caspase1 and GSDMD-N ( P<0.05); the SAH+anti-ly6G group had statistically decreased numbers of infiltrated neutrophils and FJC-positive neurons, statistically increased falling latency in the modified Garcia score and rotarod test, statistically decreased brain water content, and statistically decreased expressions of S100A8, TLR4, NLRP3, cleaved-caspase1 and GSDMD-N compared with the SAH group and SAH+vehicle group ( P<0.05). Conclusion:Inhibition of neutrophils can down-regulate the S100A8 expression after SAH and attenuate TLR4/NLRP3 activation-mediated cell pyroptosis, thereby improving EBI.

Objective To identify and verify the interacting protein of α-11 giardin, so as provide the experimental evidence for studies on the α-11 giardin function. Methods The yeast two-hybrid cDNA library of the Giardia lambia C2 strain and the bait plasmid of α-11 giardin were constructed. All proteins interacting with α-11 giardin were screened using the yeast two-hybrid system. α-11 giardin and all screened potential interacting protein genes were constructed into pBiFc-Vc-155 and pBiFc-Vn-173 plasmids, and co-transfected into the breast cancer cell line MDA-MB-231. The interactions between α-11 giardin and interacting proteins were verified using bimolecular fluorescence complementation (BiFC). Results The yeast two-hybrid G. lambia cDNA library which was quantified at 2.715 × 10⁷ colony-forming units (CFU) and the bait plasmid containing α-11 giardin gene without an autoactivation activity were constructed. Following two-round positive screening with the yeast two-hybrid system, two potential proteins interacting with α-11 giardin were screened, including eukaryotic translation initiation factor 5A (EIF5A), calmodulin-dependent protein kinase (CAMKL) and nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH), hypothetical protein 1 (GL50803_95880), hypothetical protein 2 (GL50803_87261) and a protein from Giardia canis virus. The α-11 giardin and EIF5A genes were transfected into the pBiFc-Vc-155 and pBiFc-Vn-173 plasmids using BiFC, and the recombinant plasmids pBiFc-Vc-155-α-11 and pBiFc-Vn-173-EIF5A were co-tranfected into MDA-MB-231 cells, which displayed green fluorescence under a microscope, indicating the interaction between α-11 giardin and EIF5A protein in cells. Conclusion The yeast two-hybrid cDNA library of the G. lambia C2 strain has been successfully constructed, and six potential protein interacting with α-11 giardin have been identified, including EIF5A that interacts with α-11 giardin in cells.

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.