CAR-Macrophage Therapy Alleviates Myocardial Ischemia-Reperfusion Injury.

BACKGROUND: Given the growing acknowledgment of the detrimental effects of excessive myocardial fibrosis on pathological remodeling after myocardial ischemia-reperfusion injury (I/R), targeting the modulation of myocardial fibrosis may offer protective and therapeutic advantages. However, effective clinical interventions and therapies that target myocardial fibrosis remain limited. As a promising chimeric antigen receptor (CAR) cell therapy, whether CAR macrophages (CAR-Ms) can be used to treat I/R remains unclear. METHODS: The expression of FAP (fibroblast activation protein) was studied in mouse hearts after I/R. FAP CAR-Ms were generated to target FAP-expressing cardiac fibroblasts in mouse hearts after I/R. The phagocytosis activity of FAP CAR-Ms was tested in vitro. The efficacy and safety of FAP CAR-Ms in treating I/R were evaluated in vivo. RESULTS: FAP was significantly upregulated in activated cardiac fibroblasts as early as 3 days after I/R. Upon demonstrating their ability to engulf FAP-overexpressing fibroblasts, we intravenously administered FAP CAR-Ms to mice at 3 days after I/R and found that FAP CAR-Ms significantly improved cardiac function and reduced myocardial fibrosis in mice after I/R. No toxicities associated with FAP CAR-Ms were detected in the heart or other organs at 2 weeks after I/R. Finally, we found that FAP CAR-Ms conferred long-term cardioprotection against I/R. CONCLUSIONS: Our proof-of-concept study demonstrates the therapeutic potential of FAP CAR-Ms in alleviating myocardial I/R and potentially opens new avenues for the treatment of a range of heart diseases that include a fibrotic phenotype.

Comparison of the immune response and protection against the experimental Toxoplasma gondii infection elicited by immunization with the recombinant proteins BAG1, ROP8, and BAG1-ROP8.

Toxoplasmosis is one of the most dangerous zoonotic diseases, causing serious economic losses worldwide due to abortion and reproductive problems. Vaccination is the best way to prevent disease; thus, it is imperative to develop a candidate vaccine for toxoplasmosis. BAG1 and ROP8 have the potential to become vaccine candidates. In this study, rTgBAG1, rTgROP8, and rTgBAG1-rTgROP8 were used to evaluate the immune effect of vaccines in each group by detecting the humoral and cellular immune response levels of BABL/c mice after immunization and the ability to resist acute and chronic infection with Toxoplasma gondii (T. gondii). We divided the mice into vaccine groups with different proteins, and the mice were immunized on days 0, 14, and 28. The protective effects of different proteins against T. gondii were analysed by measuring the cytokines, serum antibodies, splenocyte proliferation assay results, survival time, and number and diameter of brain cysts of mice after infection. The vaccine groups exhibited substantially higher IgG, IgG1, and IgG2a levels and effectively stimulated lymphocyte proliferation. The levels of IFN-γ and IL-2 in the vaccine group were significantly increased. The survival time of the mice in each vaccine group was prolonged and the diameter of the cysts in the vaccine group was smaller; rTgBAG1-rTgROP8 had a better protection. Our study showed that the rTgBAG1, rTgROP8, and rTgBAG1-rTgROP8 recombinant protein vaccines are partial but effective approaches against acute or chronic T. gondii infection. They are potential candidates for a toxoplasmosis vaccine.

High-performance miniature linear time-of-flight mass spectrometry as an advantageous tool in a high mass-to-charge range.

A miniature matrix-assisted laser desorption/ionization linear time-of-flight mass spectrometer suitable for the high mass-to-charge (m/z) region is described. The instrument size is roughly 1/50th that of regular instruments, and detailed dimensions and experimental parameters were optimized based on the comprehensive calculation method to provide satisfactory mass resolving power. Observations showed that the performance is limited in the low m/z range and becomes comparable with that of regular instruments in the mid m/z range. In the high m/z range, the miniature instrument provides better mass resolving power and sensitivity than regular instruments, showing superior performance for microbial, protein conjugate, and polymer analyses.

The effects of video based nursing education on perioperative anxiety and depression in patients with gastric cancer.

The purpose of this study was to determine the effect of video-based nursing education on perioperative anxiety and depression. A total of 128 patients scheduled for minimally invasive gastrectomy were randomly divided into intervention (n = 64) and control (n = 64) group. The. The anxiety and depression scores, systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) were assessed before the intervention, 1 h before surgery and 24 h after surgery. And the cortisol levels were measured before the intervention and 1 h before surgery. No significant difference was observed in baseline anxiety score, depression score, vital signs and cortisol level (P > 0.05). The anxiety level, depression level, SBP, DBP and HR of patients in intervention group was significantly lower than that in control group at 1 h before surgery and 24 hs after surgery (P < 0.05). The serum cortisol in the intervention group was also significantly lower than that in the control group 1 h before surgery (p < 0.001). Video-based nursing education was effective in decreasing the perioperative anxiety and depression of patients undergoing minimally invasive gastrectomy. It could also keep vital signs and serum cortisol levels in normal limits.

A Dynamic Data Correction Method for Enhancing Resolving Power of Integrated Spectra in Spectroscopic Analysis.

A dynamic data correction method embedded in the process of data acquisition improves spectral quality. The method minimizes the impact of random errors in spectroscopic measurements by correcting peak positions in every single-scan spectrum. The method is fast enough to facilitate online data correction. The integration of corrected spectra improves resolving power and signal-to-noise ratio. The correction method can apply to most analytical spectra. In mass spectrometry and Raman spectroscopy, observations show that it improved the average resolving power by roughly 40-150% and revealed unresolved spectral features.

Production and characterization of monoclonal antibodies against Toxoplasma gondii ROP18 with strain-specific reactivity.

The rhoptry kinase 18 of Toxoplasma gondii (TgROP18) has been identified as a key virulence factor that allows the parasite to escape from host immune defences and promotes its proliferation in host cells. Although much research is focused on the interaction between host cells and TgROP18, the development of monoclonal antibodies (mAbs) against TgROP18 has not been reported till date. To produce mAbs targeting TgROP18, two hybridomas secreting mAbs against TgROP18, designated as A1 and T2, were generated using cell fusion technology. The subtypes of the A1 and T2 mAbs were identified as IgG3 λ and IgM κ, and peptide scanning revealed that the core sequences of the antigenic epitopes were 180LRAQRRRSELVFE192 and 351NYFLLMMRAEADM363, respectively. The T2 mAb specifically reacted with both T. gondii type I and Chinese I, but not with T. gondii type II, Plasmodium falciparum or Schistosoma japonicum. Finally, the sequences of heavy chain and light chain complementarity-determining regions of T2 were amplified, cloned and characterized, making the modification of the mAb feasible in the future. The development of mAbs against TgROP18 would aid the investigation of the molecular mechanisms underlying the modulation of host cellular functions by TgROP18, and in the development of strategies to diagnose and treat Toxoplasmosis.

Hepatic ZIP8 deficiency is associated with disrupted selenium homeostasis, liver pathology, and tumor formation.

Zrt/Irt-like protein 8 (ZIP8) (encoded by Slc39a8) is a multifunctional membrane transporter that influxes essential metal cations Zn2+, Mn2+, Fe2+, and nonmetal inorganic selenite (HSeO3-). Physiological roles of ZIP8 in different cell types and tissues remain to be elucidated. We aimed to investigate ZIP8 functions in liver. Two mouse models were used in this study: 1) 13- to 21-mo-old Slc39a8(+/neo) hypomorphs having diminished ZIP8 levels and 2) a liver-specific ZIP8 acute knockdown mouse (Ad-shZip8). Histology, immunohistochemistry, and Western blotting were used to investigate ZIP8-deficiency effects on hepatic injury, inflammatory changes, and oxidative stress. Selenium (Se) and zinc (Zn) were quantified in tissues by inductively coupled plasma-mass spectrophotometry. We found that ZIP8 is required to maintain normal liver function; moderate or acute decreases in ZIP8 activity resulted in hepatic pathology. Spontaneous liver neoplastic nodules appeared in ~50% of Slc39a8(+/neo) between 13 and 21 mo of age, exhibiting features of inflammation, fibrosis, and liver injury. In Ad-shZip8 mice, significant hepatomegaly was observed; histology showed ZIP8 deficiency was associated with hepatocyte injury, inflammation, and proliferation. Significant decreases in Se, but not Zn, were found in Ad-shZip8 liver. Consistent with this Se deficit, liver expression of selenoproteins glutathione peroxidases 1 and 2 was downregulated, along with decreases in antioxidant superoxide dismutases 1 and 2, consistent with increased oxidative stress. Thus, ZIP8 plays an important role in maintaining normal hepatic function, likely through regulating Se homeostasis and redox balance. Hepatic ZIP8 deficiency is associated with liver pathology, including oxidative stress, inflammation, proliferation, and hepatocellular injury. NEW & NOTEWORTHY Zrt/Irt-like protein 8 (ZIP8) is a multifunctional membrane transporter that facilitates biometal and mineral uptake. The role of ZIP8 in liver physiology has not been previously investigated. Liu et al. discovered unique ZIP8 functions, i.e., regulation of hepatic selenium content and association of ZIP8 deficiency in mouse liver with liver defects.

High Prevalence of Erectile Dysfunction in Diabetic Men With Depressive Symptoms: A Meta-Analysis.

BACKGROUND: Erectile dysfunction (ED) may be common among diabetic men with depressive symptoms (DS), but its prevalence is still debated. AIM: To conduct a meta-analysis of the prevalence of ED in diabetic men with DS compared to those without DS, calculating the relative odds ratios (ORs) and 95% CIs. METHODS: PubMed, MEDLINE, Embase, and Web of Science were searched up to January 2018. All the studies assessing the risk of ED among diabetic men having DS were reviewed. 2 Authors independently assessed literature and extracted information eligibility. Any disagreement was resolved by a third reviewer. Newcastle-Ottawa quality assessment scale was used to evaluate study quality in meta-analyses. We calculated the ORs with 95% CIs using software Stata, Version 12.0; StataCorp, College Station, TX). Data were pooled using a fixed or random effects model according to heterogeneity. Sensitivity analyses were conducted to assess potential bias. This study was conducted according to the guidelines for Meta-Analyses and Systematic Reviews of Observational Studies. OUTCOMES: The strength of the association between DS and the prevalence of ED was evaluated using ORs and 95% CIs. RESULTS: 5 Studies were eligible for the present analysis, reporting on a total of 2525 diabetic men. Mean age of patients ranged from 42.37-61.65 years in the included studies. The overall prevalence of ED in diabetic men with DS was 74.2% (95% CI 59.0-89.4). The overall prevalence of ED in diabetic men without DS was 37.4% (95% CI 16.2-58.6). The pooled crude OR for these 5 studies was 6.40 (95% CI 2.11-19.38, P < .05, I2 = 94.6%). The pooled OR of 4 multi-variate analyses was 3.08 (95% CI 1.32-4.85, P < .001, I2 = 83.5%). CLINICAL IMPLICATIONS: Diabetic men with DS had a significantly increased prevalence of ED, suggesting that ED should be of concern to clinicians when managing diabetic men with DS. STRENGTHS & LIMITATIONS: A strength of this study is that it is the first meta-analysis to assess the prevalence of ED in diabetic men with DS and quantitatively analyze the association between DS and ED risk among diabetic men. A limitation is that all included studies were cross-sectional studies, which may generate bias. CONCLUSION: The present meta-analysis of 5 cross-sectional studies suggests that diabetic men showing DS, compared to the diabetic men without DS, have more risk of ED. Further larger prospective cohorts with more power or meta-analysis based on individual patient data need to be conducted to confirm this association. Wang X, Yang X, Cai Y, et al. High Prevalence of Erectile Dysfunction in Diabetic Men With Depressive Symptoms: A Meta-Analysis. J Sex Med 2018;15:935-941.

Toxoplasma gondii inhibits differentiation of C17.2 neural stem cells through Wnt/ß-catenin signaling pathway.

Toxoplasma gondii is a major cause of congenital brain disease. T. gondii infection in the developing fetus frequently results in major neural developmental damage; however, the effects of the parasite infection on the neural stem cells, the key players in fetal brain development, still remain elusive. This study is aiming to explore the role of T. gondii infection on differentiation of neural stem cells (NSCs) and elucidate the underlying molecular mechanisms that regulate the inhibited differentiation of NSCs induced by the infection. Using a differentiation medium, i.e. , DMEM: F12 (1:1 mixture) supplemented with 2% N2, C17.2 neural stem cells (NSCs) were able to differentiate to neurons and astrocytes, respectively evidenced by immunofluorescence staining of differentiation markers including ßIII-tubulin and glial fibrillary acidic protein (GFAP). After 5-day culture in the differentiation medium, the excreted-secreted antigens of T. gondii (Tg-ESAs) significantly down-regulated the protein levels of ßIII-tubulin and GFAP in C17.2 NSCs in a dose-dependent manner. The protein level of ß-catenin in the nucleus of C17.2 cells treated with both wnt3a (a key activator for Wnt/ß-catenin signaling pathway) and Tg-ESAs was significantly lower than that in the cells treated with only wnt3a, but significantly higher than that in the cells treated with only Tg-ESAs. In conclusion, the ESAs of T. gondii RH blocked the differentiation of C17.2 NCSs and downregulated the expression of ß-catenin, an essential component of Wnt/ß-catenin signaling pathway. The findings suggest a new mechanism underlying the neuropathogenesis induced by T. gondii infection, i.e. inhibition of the differentiation of NSCs via blockade of Wnt/ß-catenin signaling pathway, such as downregulation of ß-catenin expression by the parasite ESAs.

Macrophages suppress cardiac reprogramming of fibroblasts in vivo via IFN-mediated intercellular self-stimulating circuit.

Direct conversion of cardiac fibroblasts (CFs) to cardiomyocytes (CMs) in vivo to regenerate heart tissue is an attractive approach. After myocardial infarction (MI), heart repair proceeds with an inflammation stage initiated by monocytes infiltration of the infarct zone establishing an immune microenvironment. However, whether and how the MI microenvironment influences the reprogramming of CFs remains unclear. Here, we found that in comparison with cardiac fibroblasts (CFs) cultured in vitro, CFs that transplanted into infarct region of MI mouse models resisted to cardiac reprogramming. RNA-seq analysis revealed upregulation of interferon (IFN) response genes in transplanted CFs, and subsequent inhibition of the IFN receptors increased reprogramming efficiency in vivo. Macrophage-secreted IFN-ß was identified as the dominant upstream signaling factor after MI. CFs treated with macrophage-conditioned medium containing IFN-ß displayed reduced reprogramming efficiency, while macrophage depletion or blocking the IFN signaling pathway after MI increased reprogramming efficiency in vivo. Co-IP, BiFC and Cut-tag assays showed that phosphorylated STAT1 downstream of IFN signaling in CFs could interact with the reprogramming factor GATA4 and inhibit the GATA4 chromatin occupancy in cardiac genes. Furthermore, upregulation of IFN-IFNAR-p-STAT1 signaling could stimulate CFs secretion of CCL2/7/12 chemokines, subsequently recruiting IFN-ß-secreting macrophages. Together, these immune cells further activate STAT1 phosphorylation, enhancing CCL2/7/12 secretion and immune cell recruitment, ultimately forming a self-reinforcing positive feedback loop between CFs and macrophages via IFN-IFNAR-p-STAT1 that inhibits cardiac reprogramming in vivo. Cumulatively, our findings uncover an intercellular self-stimulating inflammatory circuit as a microenvironmental molecular barrier of in situ cardiac reprogramming that needs to be overcome for regenerative medicine applications.

Experimental Validation of Comprehensive Calculation for High-Resolution Linear MALDI-TOF Mass Spectrometry.

This work discusses the effectiveness of the previously developed comprehensive calculation model to optimize linear MALDI-TOF mass spectrometers. The model couples space- and velocity-focusing to precisely analyze the flight-time distribution of ions and predict optimal experimental parameters for the highest mass resolving power. Experimental validation was conducted using a laboratory-made instrument to analyze CsI3 and angiotensin I ions in low to medium m/z range. The results indicate that the predicted optimal extraction voltage and delay were reasonably accurate and effective. In the low m/z range, the peak width obtained using optimal parameters reached the sub nanosecond range, corresponding to a mass resolving power of 10 000-17 000, or 20 000-34 000 if shot-to-shot random fluctuations were minimized by the dynamic data correction method. The observed optimal mass resolving power in the current experiment is 4.8-7.8 times that of commercial instruments. Practical limitations resulting in the gap between the observed and theoretical ultimate mass resolving power are discussed.

Roles of carbon dioxide in the conversion of biomass or waste plastics.

Under the current trend of pursuing sustainable development and environmental protection, the important application of carbon dioxide (CO2) in the conversion process of biomass or waste plastics has become a research direction of concern. The goal of this conversion process is to achieve the efficient use of carbon dioxide, providing a process for the efficient use of biomass, and solving the environmental problems caused by plastics. Remarkable progress has been made in the study of the reaction of CO2 with other substances to produce methane, low-carbon hydrocarbons, methanol, formic acid, and its derivatives, as well as ethers, aldehydes, gasoline, low-carbon alcohols, and other chemicals. In this paper, the important role of CO2 in the conversion of alcohol, sugar, cellulose, and waste plastics was reviewed, with emphasis on the important applications of CO2 as a carbon source, reactant, reaction medium, enhancing agent, solvent, and carrier gas in the conversion of biomass or waste plastics and the basic insights of the reaction mechanism. The emerging CO2 new roles not only put forward the green application of CO2 but also have guiding significance for the utilization of biomass resources and the treatment of waste plastics.

A novel AI-based diagnostic model for pertussis pneumonia.

It is still very difficult to diagnose pertussis based on a doctor's experience. Our aim is to develop a model based on machine learning algorithms combined with biochemical blood tests to diagnose pertussis. A total of 295 patients with pertussis and 295 patients with non-pertussis lower respiratory infections between January 2022 and January 2023, matched for age and gender ratio, were included in our study. Patients underwent a reverse transcription polymerase chain reaction test for pertussis and other viruses. Univariate logistic regression analysis was used to screen for clinical and blood biochemical features associated with pertussis. The optimal features and 3 machine learning algorithms including K-nearest neighbor, support vector machine, and eXtreme Gradient Boosting (XGBoost) were used to develop diagnostic models. Using univariate logistic regression analysis, 18 out of the 27 features were considered optimal features associated with pertussis The XGBoost model was significantly superior to both the support vector machine model (Delong test, P = .01) and the K-nearest neighbor model (Delong test, P = .01), with the area under the receiver operating characteristic curve of 0.96 and an accuracy of 0.923. Our diagnostic model based on blood biochemical test results at admission and XGBoost algorithm can help doctors effectively diagnose pertussis.

Sustainable ammonia and amines from chitin.

Efforts are underway to explore alternative methods to the Haber-Bosch process for sustainable ammonia production, while the potential for ammonia extraction from natural nitrogenous biomass is under-exploited. Here, a synergistic catalytic strategy involving acid and modified Ru-based catalysts is communicated for the direct production of amines and ammonia from chitin. Phosphoric acid promotes the cleavage of ether bonds in biomass polymers and also serves to protect amino groups from being removed. Selective hydrogenation, deoxygenation, and amination can be achieved by controllably adjusting the ratio of Ru0/Run+. The utilization of nitrogen atoms in chitin can reach up to 95 % (21 % amines, 74 % ammonium), and the catalytic process is applicable to waste shrimp shells. This study demonstrates the possibility of efficient production of nitrogen-containing compounds from abundant biopolymers.

Trophoblast apoptosis through polarization of macrophages induced by Chinese Toxoplasma gondii isolates with different virulence in pregnant mice.

Toxoplasma gondii is an apicomplexan parasite capable of transplacental transmission to cause spontaneous abortion or significant disease in the surviving neonate. Different from the dominant genotypes of T. gondii strains in European and North American which belong to three distinct clonal lineages, type I, type II, and type III, isolates from China possess the predominant genotype of China 1(ToxoDB#9) with a different virulence. The genotype-associated pathogenesis has been investigated previously. Based on two isolates of T. gondii from Chinese wild cats, a murine model of pregnancy and one transwell system in vitro, here we reported differentially polarized activation of macrophages induced by genotype China 1 strains, TgCtwh3 and TgCtwh6 with different virulence to mice, and its impact on trophoblast apoptosis. The results showed that macrophages were alternatively activated when infected with virulent TgCtwh3 while classically activated when infected with low virulent (cyst-forming) TgCtwh6 both in vitro and in vivo. By the analysis of flow cytometry, the percentage of the Th1 cells in two infection groups decreased significantly, and the Th2 cells from spleen escalated only in the virulent TgCtwh3 group. Interestingly, the high parasite burden was noted in the placenta of TgCtwh3-infected group whereas the inflammatory cells infiltration predominates in the TgCtwh6-infected group. In vivo trophoblast apoptosis in TgCtwh3 group was found to be more obvious when compared with TgCtwh6 although it was present in both. The present observations indicate that polarization of macrophages and modulation of Th subsets induced by the isolates with identical genotype but different virulence could contribute to trophoblast apoptosis through different mechanisms, suggesting a virulence-associated pathogenesis of T. gondii in abnormal pregnant outcome.

Preventive effect of pidotimod on reactivated toxoplasmosis in mice.

As one of food-borne parasitic diseases, toxoplasmosis entails the risk of developing reactivation in immunocompromised patients. The synthetic dipeptide pidotimod is a potent immunostimulating agent that improves the immunodefenses in immunodepression. To investigate the efficacy of pidotimod as a preventive treatment, we used a murine model of reactivated toxoplasmosis with cyclophosphamide (CY)-induced immunosuppression. Pidotimod administration significantly restored the body weight and spleen organ index, increased survival time (from 70 to 90%), and decreased the parasitemia (from 80 to 35%) of CY-induced mice with reactivated toxoplasmosis. Cytokine profiles and CD4(+) T cells subpopulation analyses by Cytometric Bead Array and flow cytometry demonstrated that pidotimod treatment resulted in a significant upregulation of pro-inflammatory cytokines (IFN-γ, TNF-α, and IL-2) and Th1 cells (from 3.73 ± 0.39 to 5.88 ± 0.46%) after CY induction in infected mice. Additionally, histological findings and parasite DNA quantification revealed that mice administered with pidotimod had a remarkable reduction of parasite burden (two-log) and amelioration of histopathology in the brains. The in vitro studies showed that pidotimod significantly restored concanavalin A-induced splenocyte proliferation and pro-inflammatory cytokines in the supernatants of splenocyte culture. It could be concluded that the administration of pidotimod in immunocompromised mice significantly increases the Th1-biased immune response, prolongs survival time, and ameliorates the load of parasites in the blood. This is the first report of the preventive effect of pidotimod on reactivated toxoplasmosis.

Toxoplasma gondii Reactivation Aggravating Cardiac Function Impairment in Mice.

BACKGROUND: Toxoplasma gondii (T. gondii) reactivation is common, especially among immunocompromised individuals, such as AIDS patients. The cardiac involvement associated with toxoplasmosis, however, is usually obscured by neurological deterioration. The aim of this study was to observe the alterations in cardiac functions in various landmark periods after infection and to assess whether reactivation more seriously damages the heart. METHODS: We established three infection models in mice using TgCtwh6, a major strain of T. gondii prevalent in China. The groups included an acute group, chronic latent group, and reactivation group. We evaluated the cardiac function impairment via H & E staining, Masson staining, echocardiography, myocardial enzyme profiles, and cardiac troponin, and detected the expression of inflammatory factors and antioxidant factors with Western blotting. Immunofluorescence was used to detect the expression of the macrophage marker F4/80. RESULTS: Our results showed that damage to the heart occurred in the acute and reactivation groups. Impaired cardiac function manifested as a decrease in heart rate and a compensatory increase in left ventricular systolic function. Serum levels of cardiac enzymes also increased dramatically. In the chronic phase, myocardial fibrosis developed, diastolic functions became severely impaired, inflammation persisted, and macrophage expression was slightly reduced. Ultimately, reactivation infection exacerbated damage to cardiac function in mice, potentially leading to diastolic heart failure. Macrophages were strongly activated, and myocardial fibrosis was increased. In addition, the antioxidant capacity of the heart was severely affected by the infection. CONCLUSIONS: Taken together, these results suggested that the reactivation of T. gondii infection could aggravate injury to the heart, which could be associated with a host-cell-mediated immune response and strong cytokine production by macrophages, thus representing a novel insight into the pathogenic mechanism of toxoplasmosis.

Iron-overload-induced ferroptosis in mouse cerebral toxoplasmosis promotes brain injury and could be inhibited by Deferiprone.

Iron is a trace metal element that is essential for the survival of cells and parasites. The role of iron in cerebral toxoplasmosis (CT) is still unclear. Deferiprone (DFP) is the orally active iron chelator that binds iron in a molar ratio of 3:1 (ligand:iron) and promotes urinary iron excretion to remove excess iron from the body. The aims of this experiment were to observe the alterations in iron in brains with Toxoplasma gondii (T. gondii) acute infections and to investigate the mechanism of ferroptosis in CT using DFP. We established a cerebral toxoplasmosis model in vivo using TgCtwh3, the dominant strains of which are prevalent in China, and treated the mice with DFP at a dose of 75 mg/kg/d. Meanwhile, we treated the HT-22 cells with 100 µM DFP for half an hour and then infected cells with TgCtwh3 in vitro. A qRT-PCR assay of TgSAG1 levels showed a response to the T. gondii burden. We used inductively coupled plasma mass spectrometry, an iron ion assay kit, Western blot analysis, glutathione and glutathione disulfide assay kits, a malonaldehyde assay kit, and immunofluorescence to detect the ferroptosis-related indexes in the mouse hippocampus and HT-22 cells. The inflammatory factors interferon-γ, tumor necrosis factor-α, transforming growth factor-ß, and arginase 1 in the hippocampus and cells were detected using the Western blot assay. Hematoxylin and eosin staining, electron microscopy, and the Morris water maze experiment were used to evaluate the brain injuries of the mice. The results showed that TgCtwh3 infection is followed by the activation of ferroptosis-related signaling pathways and hippocampal pathological damage in mice. The use of DFP led to ferroptosis resistance and attenuated pathological changes, inflammatory reactions and T. gondii burden of the mice, prolonging their survival time. The HT-22 cells with TgCtwh3 activated the ferroptosis pathway and was inhibit by DFP in vitro. In TgCtwh3-infected cells, inflammatory response and mitochondrial damage were severe, but these effects could be reduced by DFP. Our study elucidates the mechanism by which T. gondii interferes with the host's iron metabolism and activates ferroptosis, complementing the pathogenic mechanism of CT and further demonstrating the potential value of DFP for the treatment of CT.

rROP2(186-533): a novel peptide antigen for detection of IgM antibodies against Toxoplasma gondii.

Toxoplasma gondii infections are prevalent in a wide range of mammalian hosts including humans. Infection in pregnant women may cause the transmission of parasite to the fetus that makes serious problems. IgM antibodies against Toxoplasma (Toxo-IgM) have been believed to be significant indicators for both recently acquired and congenital toxoplasmosis. So far, however, there has not been any recognized protein of T. gondii that specifically reacts to IgM antibodies. Here, an antigen exclusively for detection of IgM antibodies screened by two-dimensional electrophoresis and mass spectrometry has been reported. The study identified 13 Toxoplasma proteins probed by IgG antibodies and one (rhpotry protein 2 [ROP2]) by IgM antibodies with human sera of Toxo-IgM(-)-IgG(+) and -IgM(+)-IgG(-), respectively, which had been prescreened by Toxo-IgM and -IgG commercial kits from the suspected cases. Following cloning, expression, and purification of the fragment of ROP2(186-533), an enzyme-linked immunosorbent assay with rROP2(186-533) to measure IgM and IgG antibodies was developed. As a result, 100%(48/48) of sera with Toxo-IgM(+)-IgG(-)showed positive Toxo-IgM but none of them (0%) showed positive Toxo-IgG when rROP2(186-533) was used as antigen. Neither Toxo-IgG nor Toxo-IgM antibodies were found when tested with 59 sera of Toxo-IgM(-)-IgG(+). These results indicate that rROP2(186-533) could be used as an antigen that specifically capture Toxo-IgM antibodies and may have a high potential in the serological diagnosis of both acute acquired and congenital toxoplasmosis.

Central sterile supply department (CSSD) management quality sensitive index constructed by management mode under the guidance of key point control theory and its effect on CSSD management quality: a retrospective study.

BACKGROUND: The central sterile supply department (CSSD) ensures the quality of medical care and controls infection. The professional, standardized, and scientific management of the CSSD is prerequisite if a hospital is to realize sustainable development. Based on the management mode and under the guidance of key point control theory, this study developed the CSSD management quality sensitive index to analyze its effects on the management quality of the CSSD. METHODS: We conducted a retrospective analysis of 282 medical devices processed from January 2020 to June 2020 (the control group), and 280 medical devices sterilized from July 2020 to December 2020 (the observation group). The devices in the control group were cleaned and disinfected according to the conventional operation process, while the observation group used a management quality sensitive index that had been developed according to the management mode, as guided by the key point control theory, and managed by the CSSD. The sensitive indexes of the two groups before and after the intervention were recorded to evaluate the work quality of CSSD medical staff. RESULTS: After the intervention, the process indexes, such as the qualified rate of cleaning quality, qualified rate of assembly, qualified rate of labeling, and correct rate of sterilization, the qualification rate of the sterilization results, the rate of intact packaging, and the rate of intact instruments, the environmental status, packaging quality, cleaning quality, and equipment management scores of the observation group were significantly higher than those of the control group (P<0.05). The incidence of wet packaging in the observation group was significantly lower than that in the control group (P<0.05). The service quality indexes, such as the replenishment time, retrieval time and preparation time of goods, of the observation group were significantly shorter than those of the control group (P<0.05). CONCLUSIONS: The quality sensitive index constructed under the guidance of the key point control theory can be used to guide the quality management of the CSSD, improve the processing results of key points in key control processes, and improve the work quality and service quality of the medical staff.