Beneficial Effect of Toxoplasma gondii Infection on the Prognosis of Breast Cancer Was Modified by Cytokines.

Background: Animal experiments have shown the anticancer activity of Toxoplasma gondii (T. gondii), but its effect on the prognosis of cancer patients is unclear. Thus, the present study aimed to investigate the prognostic role of anti-T. gondii IgG in breast cancer patients and the modification effect of cytokines. Methods: A total of 1121 breast cancer patients were recruited between 2008 and 2018 and followed up until December 31, 2021. Anti-T. gondii IgG and cytokines were measured using an enzyme-linked immunosorbent assay (ELISA) kit and a multiplex assay platform. Endpoints were overall survival (OS) and progression-free survival (PFS). Survival and multiplicative interaction analyses were performed using multivariate Cox regression models. Results: According to the cutoff value of optical density (OD=0.111), 900 (80.29%) and 221 (19.71%) patients were divided into two groups: low or high anti-T. gondii IgG. Compared to patients with a low anti-T. gondii IgG level, the adjusted hazard ratios (HRs) of OS and PFS for patients with high anti-T. gondii IgG levels were 0.60 (95% confidence interval (CI): 0.37-0.99) and 0.67 (0.46-0.98), respectively. These associations were profound among patients with a high cytokine score (HR=0.29, 95% CI: 0.10-0.82 for OS; HR=0.30, 95% CI: 0.13-0.69 for PFS), accompanied by a significant interaction between the level of anti-T. gondii IgG and the cytokine score (P interaction=0.019 for PFS); interleukin-17 (IL-17) and interleukin-9 (IL-9) were the main contributors to the interaction. Conclusion: Anti-T. gondii IgG was found to be beneficial to breast cancer survival, especially in women with systematic inflammation and high IL-17 or IL-9 levels, suggesting the potential of T. gondii as a prognostic marker and a novel immunotherapy approach for cancer patients.

Subendocardial Involvement as an Underrecognized Cardiac MRI Phenotype in Myocarditis.

Background Subendocardial late gadolinium enhancement (LGE) detected with cardiac MRI in myocarditis represents a diagnostic dilemma, since it may resemble myocardial ischemia. Purpose To explore and compare the histopathologic characteristics and clinical features and outcomes in patients with myocarditis with and without subendocardial involvement at cardiac MRI. Materials and Methods This retrospective study evaluated 39 patients with myocarditis pathologically proven by means of either endomyocardial biopsy or explant pathologic findings between 2015 and 2020. Patients were divided into two groups according to cardiac MRI phenotype: 18 with subendocardial involvement (mean age ± standard deviation, 40 years ± 17; 10 women) and 21 with no subendocardial involvement (mean age, 35 years ± 11; six women). The median follow-up period was 784 days (interquartile range [IQR], 90-1123 days). The Student t test, Mann-Whitney U test, and univariable Cox regression were used for statistical analyses. Results In the 18 patients with subendocardial involvement, 12 (67%) had lymphocytic myocarditis and six (33%) had giant cell myocarditis. Patients with subendocardial involvement compared with those without subendocardial involvement had lower left ventricular ejection fraction (mean ± standard deviation, 27% ± 11 vs 41% ± 19; P = .004), larger LGE extent (median, 13% [IQR, 10%-22%] vs 5% [IQR, 2%-17%]; P < .001), higher rates of cardiac death or transplant (eight of 18 patients [44%] vs one of 21 patients [4.8%]; P = .006), higher probability of giant cell myocarditis (six of 18 [33%] vs one of 21 [4.8%]; P = .02), and more major adverse cardiovascular events (MACE) (15 of 18 [83%] vs seven of 21 [33%]; P = .002). In a subgroup of patients with comparable LGE extent (median, 15% vs 16%; P = .40) and left ventricular ejection fraction (median, 27% vs 31%; P = .26), the prognostic difference in terms of MACE remained (15 of 17 patients [88%] vs five of 10 [50%]; P = .02). Conclusion Subendocardial involvement detected with cardiac MRI in myocarditis indicated more severe clinical features, including a higher frequency of severe lymphocytic myocarditis or giant cell myocarditis and worse prognosis. © RSNA, 2021 See also the editorial by de Roos in this issue.

Strengthening effects of bone marrow mononuclear cells with intensive atorvastatin in acute myocardial infarction.

OBJECTIVE: To test whether intensive atorvastatin (ATV) increases the efficacy of transplantation with autologous bone marrow mononuclear cells (MNCs) in patients suffering from anterior ST-elevated myocardial infarction (STEMI). METHODS: This clinical trial was under a 2×2 factorial design, enrolling 100 STEMI patients, randomly into four groups of regular (RA) or intensive ATV (IA) with MNCs or placebo. The primary endpoint was the change of left ventricular ejection fraction (LVEF) at 1-year follow-up from baseline, primarily assessed by MRI. The secondary endpoints included other parameters of cardiac function, remodelling and regeneration determined by MRI, echocardiography, positron emission tomography (PET) and biomarkers. RESULTS: All the STEMI patients with transplantation of MNCs showed significantly increased LVEF change values than those with placebo (p=0.01) with only in the IA+MNCs patients group demonstrating significantly elevation of LVEF than in the IA+placebo group (+12.6% (95%CI 10.4 to 19.3) vs +5.0% (95%CI 4.0 to 10.0), p=0.001), pointing to a better synergy between ATV and MNCs (p=0.019). PET analysis revealed significantly increased viable areas of myocardium (p=0.015), while the scar sizes (p=0.026) and blood aminoterminal pro-B-type natriuretic peptide (p<0.034) reduced. All these above benefits of MNCs were also attributed to IA+MNCs instead of RA+MNCs group of patients with STEMI. CONCLUSIONS: Intensive ATV treatment augments the therapeutic efficacy of MNCs in patients with anterior STEMI at the convalescent stage. The treatment with the protocol of intensive ATV and MNC combination offers a clinically essential approach for myocardial infarction. TRIAL REGISTRATION NUMBER: NCT00979758.

Transplantation efficacy of autologous bone marrow mesenchymal stem cells combined with atorvastatin for acute myocardial infarction (TEAM-AMI): rationale and design of a randomized, double-blind, placebo-controlled, multi-center, Phase II TEAM-AMI trial.

Aim: To determine the efficacy and safety of intracoronary infusion of autologous bone marrow mesenchymal stem cells (MSCINJ) in combination with intensive atorvastatin (ATV) treatment for patients with anterior ST-segment elevation myocardial infarction-elevation myocardial infarction. Patients & methods: The trial enrolls a total of 100 patients with anterior ST-elevation myocardial infarction. The subjects are randomly assigned (1:1:1:1) to receive routine ATV (20 mg/d) with placebo or MSCsINJ and intensive ATV (80 mg/d) with placebo or MSCsINJ. The primary end point is the absolute change of left ventricular ejection fraction within 12 months. The secondary end points include parameters in cardiac function, remodeling and regeneration, quality of life, biomarkers and clinical outcomes. Results & conclusion: The trial will implicate the essential of cardiac micro-environment improvement ('fertilizing') for cell-based therapy. Clinical Trial Registration: NCT03047772.

Myocardial scar identified by magnetic resonance imaging can predict left ventricular functional improvement after coronary artery bypass grafting.

BACKGROUND: Previous studies have shown that viable myocardium predicts recovery of left ventricular (LV) dysfunction after revascularization. Our aim was to evaluate the prognostic value of myocardial scar assessed by late gadolinium-enhanced cardiovascular magnetic resonance imaging (LGE-CMR) on functional recovery in patients undergoing coronary artery bypass grafting (CABG). METHODS: From November 2009 to September 2012, 63 patients with reduced left ventricular ejection fraction (LVEF) referred for first-time isolated CABG were prospectively enrolled, 52 were included in final analysis. LV functional parameters and scar tissue were assessed by LGE-CMR at baseline and 6 months after surgery. Patency of grafts was evaluated by computed tomography angiography (CTA) 6 months post-CABG. Predictors for global functional recovery were analyzed. RESULTS: The baseline LVEF was 32.7 ± 9.2%, which improved to 41.6 ± 11.0% 6 months later and 32/52 patients improved LVEF by ≥ 5%. Multivariate logistic regression analysis showed that the most significant negative predictor for global functional recovery was the number of scar segments (Odds ratio 2.864, 95% Confidence Interval 1.172-6.996, p = 0.021). Receiver-Operator-Characteristic (ROC) analysis demonstrated that ≤ 4 scar segments predicted global functional recovery with a sensitivity and specificity of 85.0% and 87.5%, respectively (AUC = 0.91, p<0.001). Comparison of ROC curves also indicated that scar tissue was superior to viable myocardium in predicting cardiac functional recovery (p<0.001). CONCLUSIONS: Our findings indicated that scar tissue on LGE-CMR is an independent negative predictor of cardiac functional recovery in patients with impaired LV function undergoing CABG. These observations may be helpful for clinicians and cardiovascular surgeons to determine which patients are most likely to benefit from surgical revascularization.

Atorvastatin enhance efficacy of mesenchymal stem cells treatment for swine myocardial infarction via activation of nitric oxide synthase.

BACKGROUND: In a swine model of acute myocardial infarction (AMI), Statins can enhance the therapeutic efficacy of mesenchymal stem cell (MSCs) transplantation. However, the mechanisms remain unclear. This study aims at assessing whether atorvastatin (Ator) facilitates the effects of MSCs through activation of nitric oxide synthase (NOS), especially endothelial nitric oxide synthase (eNOS), which is known to protect against ischemic injury. METHODS AND RESULTS: 42 miniswines were randomized into six groups (n = 7/group): Sham operation; AMI control; Ator only; MSC only, Ator+MSCs and Ator+MSCs+NG-nitrol-L-arginine (L-NNA), an inhibitor of NOS. In an open-heart surgery, swine coronary artery ligation and reperfusion model were established, and autologous bone-marrow MSCs were injected intramyocardium. Four weeks after transplantation, compared with the control group, Ator+MSCs animals exhibited decreased defect areas of both "perfusion" defined by Single-Photon Emission Computed Tomography (-6.2±1.8% vs. 2.0±5.1%, P = 0.0001) and "metabolism" defined by Positron Emission Tomography (-3.00±1.41% vs. 4.20±4.09%, P = 0.0004); Ejection fraction by Magnetic Resonance Imaging increased substantially (14.22±12.8% vs. 1.64±2.64%, P = 0.019). In addition, indices of inflammation, fibrosis, and apoptosis were reduced and survivals of MSCs or MSC-derived cells were increased in Ator+MSCs animals. In Ator or MSCs alone group, perfusion, metabolism, inflammation, fibrosis or apoptosis were reduced but there were no benefits in terms of heart function and cell survival. Furthermore, the above benefits of Ator+MSCs treatment could be partially blocked by L-NNA. CONCLUSIONS: Atorvastatin facilitates survival of implanted MSCs, improves function and morphology of infarcted hearts, mediated by activation of eNOS and alleviated by NOS inhibitor. The data reveal the cellular and molecular mechanism for anti-AMI therapy with a combination of statin and stem cells.

[In vivo re-distribution of intra-coronary transplanted stem cells into beating and arrested hearts by magnetic resonance imaging in an acute myocardial infarction swine model].

OBJECTIVE: To analyze the stem cell re-distribution after intra-coronary infusion (ICI) into arrested and beating hearts in a swine myocardial infarction (MI) model using magnetic resonance imaging (MRI). METHODS: Bone marrow-derived mesenchymal stem cells were obtained from male swine and labeled with iron oxide during culture. One week after MI in female swine, the survivors were randomly divided into 4 groups. Cardiopulmonary bypass was set up to arrest the heart, and then SPIO labeled male stem cells (1 × 10(8)) were infused through coronary of beating heart (n = 6) and the arrested heart (n = 6). Saline was injected in either the beating or arresting heart as respective controls. Three days later, cell distribution was assessed by T2(*) change with magnetic resonance imaging and Y-chromosome (SRY) was detected with quantitative polymerase chain reaction. RESULTS: The reduction of T2(*) values was significantly different in the hearts, spleens, livers and lung between the transplantation groups and the control groups. Only few transplanted cells were localized in the heart and T2(*) values were similar between beating and arrest heart groups [(-7.81 ± 2.03) ms vs. (-6.56 ± 1.72) ms, P > 0.05], while T2(*) value reduction was more significant in the spleen and liver in arrest heart group than in beating heart group [spleen: (-16.72 ± 2.83) ms vs. (-22.18 ± 3.98) ms, P < 0.01, liver: (-2.40 ± 0.44) ms vs. (-5.32 ± 3.40) ms, P < 0.05]. T2(*) value was similar in kidney among the four groups. qRT-PCR detected SRY gene was similar in the heart, less in the spleen and liver while more in the lung in beating heart group compared to arrested heart group. In vitro Prussian blue stained positively transplanted cells were found in the above organs in transplantation group. CONCLUSIONS: The majority of stem cells transplanted by ICI would be entrapped by the extracardiac organs. Stem cell transplantation via ICI into the arrested heart does not favor more cells retention in the injured myocardium. Further investigation is needed to optimize the approach of stem cell delivery.

[Gelatin microspheres containing vascular endothelial growth factor enhances the efficacy of bone marrow mesenchymal stem cells transplantation in a swine model of myocardial infarction.].

OBJECTIVE: To investigate the efficacy of transplantation of mesenchymal stem cells (MSC) with gelatin microspheres containing vascular endothelial growth factor in ischemic regions in infracted swine hearts. METHODS: Twelve Chinese mini swines with infarction were randomized to receive autogenetic MSC injection to the peri-infarction area of left ventricular wall (MSC group, n = 6) or MSC transplantation with gelatin hydrogel microspheres incorporating vascular endothelial growth factor (VEGF-MSC group, n = 6). Three weeks later, left ventricular function was assessed by magnetic resonance imaging (MRI). The contrast of the MSC hypointense lesion was determined using the difference in signal intensity between the hypointense and normal myocardium divided by signal intensity of the normal region. Myocardial capillary density, the number of DAPI positive MSC and the apoptotic MSC were also determined. RESULTS: The diameter of the microspheres averaged (104.0 +/- 22.6) microm. At 24 hours after transplantation, MSC were identified by MRI as large intramyocardial signal voids at injection sites which persisted up to 3 weeks. There was no significant difference in the contrast of the lesions and in the size of the lesions at 24 hours between two groups. At 3 weeks after injection, the size of the lesions and the contrast of the lesion were decreased (P < 0.05) in both groups. The capillary density of the injection site was significantly more in the MSC-VEGF microsphere group than that in MSC group [(15.2 +/- 5.4)/HPF vs. (10.2 +/- 5.0)/HPF, t = 2.43, P < 0.05], and there were more dense DAPI labeled MSC per high power fields in injection sites of MSC-VEGF microsphere group than that in MSC group [(354 +/- 83)/HPF vs. (278 +/- 97)/HPF, t = 3.14, P < 0.05]. Moreover, the apoptosis rate of MSCs of MSCs-VEGF microsphere group was less than that of MSC group [(6.4 +/- 4.1)% vs. (11.9 +/- 4.8)%, t = 2.97, P < 0.05]. CONCLUSIONS: MSC transplantation with gelatin hydrogel microspheres incorporating VEGF enhanced the efficacy of MSC in this swine model of myocardial infarction. MRI tracking of MSC is feasible and represents a preferred method for studying the engraftment of MSCs in infracted tissue.

[Assessment of therapeutic effects of stem cell transplantation in heart failure patients with old myocardial infarction by magnetic resonance imaging].

OBJECTIVE: To evaluate the therapeutic effects of stem cell transplantation in heart failure patients with old myocardial infarction (OMI) by MRI. METHODS: Heart failure patients [NYHA 2.7 +/- 0.7, male = 18, mean age (59.5 +/- 10.1) y] with OMI were randomly divided into 2 groups (group A: CABG + stem cell transplantation, group B: CABG; n = 10 each). Left ventricular (LV) function was measured by MRI, viable myocardium was detected by (18)F-FDG myocardial metabolism imaging and late contrast-enhanced at baseline and 6 months post intervention. RESULTS: LVEF and LVEDV at baseline for group A were (20.71 +/- 6.09)% and (172.73 +/- 32.74) ml, and for group B were (27.59 +/- 2.31)% and (155.13 +/- 28.36) ml, respectively (P > 0.05). The LVEF was equally improved in group A and B (mean 8.63% vs. 10.37%, P > 0.05) while DeltaLVEDV was significant higher in group A than that in group B [(9.91 +/- 39.50) ml vs. (-22.34 +/- 31.35) ml, P < 0.05]. Ventricular wall thickening ratio at 6 months post intervention was significantly higher in group A than that in group B [(11.40 +/- 11.53)% vs. (2.27 +/- 7.20)%, P < 0.05]. Late contrast-enhanced MRI results correlated with (18)F-FDG myocardial metabolism imaging SPECT well in assessment of myocardial viability (kappa value: 0.446, P < 0.001; sensitivity: 68.3% and specificity: 92.5%). CONCLUSIONS: Stem cell therapy on top of CABG aggravated LV remodeling in heart failure patients with old myocardial infarction. The specificity of MRI is similar to (18)F-FDG SPECT while the sensitivity is inferior to (18)F-FDG SPECT on detecting viable myocardium.

Atorvastatin treatment improves survival and effects of implanted mesenchymal stem cells in post-infarct swine hearts.

AIMS: To investigate whether Atorvastatin (Ator) treatment improves the cardiac micro-environment that facilitates survival and differentiation of bone-marrow-derived mesenchymal stem cells (MSCs) implanted in the post-infarct myocardium. METHODS AND RESULTS: Myocardial infarction was created by coronary ligation and immediately after reperfusion, autologous bone-marrow-derived MSCs were transplanted into the hearts of Chinese swine that were pretreated with or without Ator. Six weeks after transplantation, as evaluated by SPECT and MRI all the animals with Ator showed improved cardiac perfusion and contractility when compared with untreated. Increased survival and differentiation of implanted MSCs and decreased infarct area were observed in the Ator-treated, MSC-implanted animals. In the absence of Ator, MSC transplantation only achieved a modest improvement in perfusion and morphology. The combined treatment with Ator and MSCs significantly inhibited cardiac cell apoptosis, reduced oxidative stress, and suppressed expression of the inflammatory cytokines in the post-infarct myocardium. CONCLUSION: Ator treatment may protect the myocardium undergoing acute infarction and reperfusion by creating a better environment for the survival and differentiation of implanted MSCs. The benefit of the Ator/stem cell combined therapy may result from the statin-mediated inhibition of apoptosis, oxidative stress, and inflammation in the infarcted myocardium.

Effects of Tongxinluo-facilitated cellular cardiomyoplasty with autologous bone marrow-mesenchymal stem cells on postinfarct swine hearts.

BACKGROUND: Treatment of ischemic heart disease remains an important challenge, though there have been enormous progresses in cardiovascular therapeutics. This study was conducted to evaluate whether Tongxinluo (TXL) treatment around the transplantation of mesenchymal stem cells (MSCs) can improve survival and subsequent activities of implanted cells in swine hearts with acute myocardial infarction (AMI) and reperfusion. METHODS: Twenty-eight Chinese mini-pigs were divided into four groups including a control group (n = 7); group 2, administration of low-dose TXL alone from the 3rd day prior to AMI to the 4th day post transplantation (n = 7); group 3, MSCs alone (n = 7) and group 4, TXL + MSCs (n = 7). AMI models were made by occlusion of the left anterior descending coronary artery for 90 minutes. Autologous bone marrow-MSCs (3 x 10(7) cells/animal) were then injected into the post-infarct myocardium immediately after AMI and reperfusion. The survival and differentiation of implanted cells in vivo were detected by immunofluorescent analysis. The data of cardiac function were obtained at baseline (1 week after transplantation) and endpoint (6 weeks after transplantation) by single photon emission computed tomography (SPECT) and magnetic resonance imaging (MRI). Apoptosis was detected by TUNEL assay and the oxidative stress level was investigated in the post-infarct myocardium at endpoint. RESULTS: At endpoint, there was less fibrosis and inflammatory cell infiltration with more surviving myocardium in group 4 than in the control group. In group 4 the survival and differentiation of implanted MSCs were significantly improved more than that seen in group 3 alone (P < 0.0001); the capillary density was also significantly greater than in the control group, group 2 or 3 both in the infarcted zone (P < 0.0001) and the peri-infarct zone (P < 0.0001). MRI showed that parameters at baseline were not significantly different between the 4 groups. At endpoint, regional wall thickening and the left ventricular ejection fraction were increased while the left ventricular mass index, dyskinetic segments and infarcted size were decreased only in group 4 compared with control group (P < 0.0001). SPECT showed that the area of perfusion defect was significantly decreased at endpoint only in group 4 compared with control group (P < 0.0001). TUNEL assay indicated that TXL administration significantly decreased cell apoptosis in peri-infarct myocardium in groups 2 and 4. Furthermore, superoxide dismutase (SOD) significantly increased and malondialdehyde (MDA) decreased in groups 2 and 4 by the administration of TXL. CONCLUSIONS: Our study demonstrates the following: (1) immediate intramyocardial injection of MSCs after AMI and reperfusion resulted in limited survival and differentiation potential of implanted cells in vivo, thus being incapable of beneficially affecting post-hearts; (2) TXL-facilitation resulted in a significant survival and differentiation potential of implanted cells in vivo via inhibition of apoptosis and oxidative stress, accompanied by significant benefits in cardiac function.

[Diagnostic value of magnetic resonance imaging for arrhythmogenic right ventricular cardiomyopathy].

OBJECTIVE: To evaluate the diagnostic value of magnetic resonance imaging (MRI) for arrhythmogenic right ventricular cardiomyopathy (ARVC). METHODS: MRI was performed in 27 (male 21, mean age: 37.4 y, ranging from 15 - 67 y) clinically diagnosed ARVC patients according to the 1994 ARVC diagnosis criteria of WHO from Oct. 2004 to Jun. 2006. Heart chamber size, fat infiltration, local or global ventricular function, myocardium perfusion of contrast first pass and late enhancement were examined. RESULTS: Fat infiltration was found in 24 (88.89%), trabecular disarray in 17 (62.96%), significant dilated right ventricle outlet (RVOT) in 18 (66.67%), dilated RV apex in 14 (51.85%), dilated RV free wall and posterior wall in 18 (66.67%) and right atrium enlargement in 11 (40.74%) patients. Local RV dysfunction was found in 18.52% (5/27), global RV dysfunction in 70.37% (19/27) of patients with mean RV ejection fraction (EF) of 35%. Left ventricle was affected in 40.74% (11/27) of patients. Perfusion defects were found in only 10.52% (2/19) of patients. Positive late enhancement of myocardium were found in 36.84% (7/19) of patients and affecting mainly the wall of RVOT and the free wall associated with lateral wall enhancement of LV. Five patients received heart transplantation and histology on transplanted hearts confirmed the MRI findings. CONCLUSION: "One-stop-shop" MRI scanning can be used for the diagnosis of ARVC. While for some ARVC cases with dominant abnormality in LV, it is difficult for MRI to differentiate ARVC from dilated cardiomyopathy or coronary heart disease. We found fibrosis of lateral wall of LV can be a characteristic sign of ARVC.