Vinblastine resets tumor-associated macrophages toward M1 phenotype and promotes antitumor immune response.

BACKGROUND: Massive tumor-associated macrophage (TAM) infiltration is observed in many tumors, which usually display the immune-suppressive M2-like phenotype but can also be converted to an M1-like antitumor phenotype due to their high degree of plasticity. The macrophage polarization state is associated with changes in cell shape, macrophage morphology is associated with activation status. M1 macrophages appeared large and rounded, while M2 macrophages were stretched and elongated cells. Manipulating cell morphology has been shown to affect the polarization state of macrophages. The shape of the cell is largely dependent on cytoskeletal proteins, especially, microtubules. As a microtubule-targetting drug, vinblastine (VBL) has been used in chemotherapy. However, no study to date has explored the effect of VBL on TAM shape changes and its role in tumor immune response. METHOD: We used fluorescent staining of the cytoskeleton and quantitative analysis to reveal the morphological differences between M0, M1, M2, TAM and VBL-treated TAM. Flow cytometry was used to confirm the polarization states of these macrophages using a cell surface marker-based classification. In vivo antibody depletion experiments in tumor mouse models were performed to test whether macrophages and CD8+ T cell populations were required for the antitumor effect of VBL. VBL and anti-PD-1 combination therapy was then investigated in comparison with monotherapy. RNA-seq of TAM of treated and untreated with VBL was performed to explore the changes in pathway activities. siRNA mediated knockdown experiments were performed to verify the target pathway that was affected by VBL treatment. RESULTS: Here, we showed that VBL, an antineoplastic agent that destabilizes microtubule, drove macrophage polarization into the M1-like phenotype both in vitro and in tumor models. The antitumor effect of VBL was attenuated in the absence of macrophages or CD8+ T cells. Mechanistically, VBL induces the activation of NF-κB and Cyba-dependent reactive oxygen species generation, thus polarizing TAMs to the M1 phenotype. In parallel, VBL promotes the nuclear translocation of transcription factor EB, inducing lysosome biogenesis and a dramatic increase in phagocytic activity in macrophages. CONCLUSIONS: This study explored whether manipulating cellular morphology affects macrophage polarization and consequently induces an antitumor response. Our data reveal a previously unrecognized antitumor mechanism of VBL and suggest a drug repurposing strategy combining VBL with immune checkpoint inhibitors to improve malignant tumor immunotherapy.

Correlation of electroencephalogram background evolution with the degree of brain injury in neonates with hypoxic-ischemic encephalopathy. / æ–°ç”Ÿå„¿ç¼ºæ°§ç¼ºè¡€æ€§è„‘ç— è„‘ç”µèƒŒæ™¯æ¼”å˜ä¸Žè„‘æŸä¼¤ç¨‹åº¦çš„ç›¸å ³æ€§ç ”ç©¶.

OBJECTIVES: To study the correlation of electroencephalogram (EEG) background evolution with the degree of brain injury in neonates with hypoxic-ischemic encephalopathy (HIE). METHODS: A retrospective analysis was performed for 56 neonates with HIE who underwent continuous video electroencephalogram (cVEEG) and brain magnetic resonance imaging (MRI) examinations. According to clinical symptoms, they were divided into a mild group with 3 neonates, a moderate group with 36 neonates, and a severe group with 17 neonates. EEG background grading and MRI score were determined for each group to analyze the correlation of EEG background evolution with the degree of brain injury. RESULTS: Compared with the moderate group, the severe group had significantly lower gestational age and Apgar score at 5 minutes after birth, a significantly higher resuscitation score, significantly lower base excess in umbilical cord blood or blood gas within 1 hour, a significantly higher proportion of neonates on mechanical ventilation, and a significantly higher incidence rate of short-term adverse outcomes (P<0.05). For the neonates in the mild and moderate groups, MRI mainly showed no brain injury (67%, 2/3) and watershed injury (67%, 16/24) respectively, and EEG showed mild abnormality in 62% (13/21) of the neonates on the 3rd day after birth. For the neonates in the severe group, MRI mainly showed basal ganglia/thalamus + brainstem injury (24%, 4/17) and whole brain injury (71%, 12/17), and EEG showed moderate or severe abnormalities on the 3rd day after birth. EEG background grading was correlated with clinical grading, MRI score, and short-term outcome on days 1, 2, 3 and 7-14 after birth (P<0.01). The highest correlation coefficient between EEG grading and MRI score was observed on the 3rd day after birth (rs=0.751, P<0.001), and the highest correlation coefficients between EEG grading and clinical grading (rs=0.592, P=0.002) and between EEG grading and short-term outcome (rs=0.737, P<0.001) were observed 7-14 days after birth. Among the neonates with severe abnormal EEG, the neonates without brain electrical activity had the highest MRI score, followed by those with status epileptics and persistent low voltage (P<0.05). CONCLUSIONS: There is a good correlation between EEG background grading and degree of brain injury in neonates with HIE, which can help to evaluate the degree and prognosis of brain injury in the early stage.

[Assessment of Left Atrial Appendage Size and Morphology by Enhanced Cardiac Computed Tomography in Patients with Non-valvular Atrial Fibrillation].

OBJECTIVE: This study was designed to investigate left atrial appendage (LAA) size and morphology characteristics in patients with non-valvular atrial fibrillation (NVAF) by enhanced cardiac computed tomography (CT) scanning,and to evaluate the predictive value of these parameters for thromboembolic risk. METHODS: A total of 189 NVAF patients were prospectively enrolled during December 2012 to January 2014 in West China Hospital of Sichuan University. Fifty-two atrial tachycardia patients without organ dysfunction were also recruited as controls. All subjects were scheduled for radiofrequency ablation and underwent enhanced cardiac CT scan. The clinical characteristics,comorbid diseases,anticoagulant therapy,and AF chronicity were collected and CHA2DS2-VASc score was calculated for the assessment of thromboembolic risk. The NVAF cohort was also divided further into LAA thrombus present (n=13) and absent (n=176) subgroup according to LAA with thrombus or not. Differencesin size and morphology characteristics of the LAA were compared among controls and different CHA2DS2-VASc thromboembolic risk subgroups,and between LAA thrombus and non-thrombus group. In addition,logistic regression analysis was used to identify whether these parameters predict thrombus formation independent of the CHA2DS2-VASc score in NVAF patients. RESULTS: Compared with controls,NVAF patients had significantly bigger orifice size. Windsock and cactus-type LAA were main morphology in controls,while the distribution of the four kinds of LAA types were similar in NVAF patients. The orifice and volume of the LAA were increased with increasing CHA2DS2-VASc riskscore,and chicken wing-type LAA was the predominant prevalence in low-intermediate thromboembolic risk subgroup while cauliflower in high thromboembolic risk subgroup. Compared with non-thrombus patients,LAA-thrombussubgroup exhibited a significantly increased orifice and volume,and higher prevalenceof the cauliflower shaped LAA. Logistic regression showed that the LAA orifice and cauliflower shape were independent predictors for thrombus formation in patients with NVAF following adjustment for the CHA2DS2-VASc score. CONCLUSION: Cardiac images obtained with CTis feasible for the analysis of LAA size and morphology,and these parameters provide incremental value in assessing the LAA thrombus risk in patients with NVAF.

Dissipative energy loss within the left ventricle detected by vector flow mapping in diabetic patients with controlled and uncontrolled blood glucose levels.

Diabetes mellitus (DM) is related to increased risks of cardiovascular diseases, such as myocardial infarction, diabetic cardiomyopathy and secondary hypertension. Dissipative energy loss (EL) derived from vector flow mapping (VFM) is thought to reflect the efficiency of blood flow and has been deemed to be an index for the evaluation of left ventricular function. Our study aimed to investigate the value of dissipative EL in diabetic patients with controlled and uncontrolled blood glucose by VFM. Eighty-eight patients with DM and 58 age-matched healthy controls were recruited. All of the patients received echocardiography examinations. VFM analyses were executed to calculate the EL values according to the apical four-chamber examinations from the left ventricle (LV) view. Our results showed that diastolic EL was compromised in the controlled-blood glucose (59.19 mV/m vs. 32.68 mV/m, p = 0.039) patients and was more dramatically increased in the uncontrolled blood glucose group (88.84 mV/m vs. 32.68 mV/m, p < 0.001) compared with the healthy controls. The impairment of systolic EL was observed only in the uncontrolled blood glucose patients (39.65 mV/m vs. 20.29 mV/m, p < 0.001) and not in the controlled blood glucose patients (29.25 mV/m vs. 20.29 mV/m, p = 0.072). Multivariate backward stepwise linear regression analysis revealed that the HbA1c level was independently related to the diastolic EL (ß = 0.233, p = 0.026) and systolic EL (ß = 0.237, p = 0.023). VFM is feasible and reproducible for assessing LV dissipative EL in DM patients with normal LVEF values in whom diastolic EL may be a more vulnerable indicator of early LV cardiac dysfunction in patients with DM. However, LV systolic EL may be a sensitive indicator of preclinical LV dysfunction for patients with DM with uncontrolled blood glucose levels. Uncontrolled blood glucose, which is independently correlated with subclinical LV dysfunction, may lead to increases in systolic EL and diastolic EL in LV.

[Aortic and Mitral Annuli Coupling and Its Association with Cardiac Function in Patients with Dilated Cardiomyopathy.]

OBJECTIVES: To compare the mitral and aortic annuli excursion (AE) and aortomitral angle (AMA) variations between patients with and without dilated cardiomyopathy (DCM) using speckle tracking echocardiography (STE),and determine the role of annuli dynamics and coupling behaviors in the left ventricular function. METHODS: Thirty-five patients with DCM and thirty-five age and sex matched healthy adults underwent transthoracic echocardiographic examinations.In the parasternal long-axis view,the AE and AMA variations were measured using Philips QLAB 9.0 software.In the apical four chamber view,the left ventricular ejection fraction (LVEF),longitudinal,circumferential,and radial strain were measured using Tomtec Arene 1.0 software. RESULTS: The DCM patients had significantly smaller AE and AMAmax-min,bigger AMAmax and AMAmin,and varied cardiac cycle compared with the healthy adults.The AMA of DCM patients increased to its maximal value earlier in diastole with little variation.AE was positively correlated with LVEF and multidirectional strain.AMAmax and AMAmin were negatively correlated with AE,LVEF and multidirectional strain (P<0.05).Significant correlations were also found between AMAmax-min and AE,LVEF and multidirectional strain (P<0.05). CONCLUSIONS: AE,AMA,LVEF and multidirectional strain can be quantified using STE.The coupling of mitral and aortic annuli in DCM patients is abnormal and correlated well with left ventricular systolic function.AE and AMA can be conveniently used for estimation of integral and regional cardiac function.

[Real-time Three-dimensional Dobutamine Stress Echocardiography for the Evaluation of Ischemic Mitral Regurgitation].

OBJECTIVE: To evaluate mitral regurgitation (MR) and left ventricular myocardial function in the patients with ischemic mitral regurgitation (IMR) by real-time three-dimensional dobutamine stress echocardiography (RT3D-DSE). METHODS: Seventy two consecutive in-patients with myocardial infarction and MR were enrolled in the study during 2006-2012. Each patient underwent RT3D-DSE examination before treatment. Three-dimensional (3D) heart full-volume data and 3D MR were obtained at baseline and each stress state. Quantitative parameters including: left ventricular end-diastolic/systolic volume (LVEDV/LVESV), left ventricular ejection fraction (LVEF) , left ventricular diastolic /systolic mass (LVMd/s). The time to the point of minimal regional systolic volume (Tmsv) was measured from the time volume curves of 17 segments, the standard deviation (Tmsv-SD) and maximal difference (Tmsv-Dif) of Tmsv were calculated. Excursion max (Emax), excursion SD (Esd), vena contarcta area (VCA) also were measured. RESULTS: 94.4% patients reached dobutamine (Dob) at 30 µg/(kg · min), 69.4% patients completed Dob 40 µg/(kg · min), only 38.9% patients completed Dob at 50 µg/(kg · min). At different stress state, VCA decreased significantly, and the decrement was up to the largest at 30 µg/(kg · min). 2D-LVEF, 3D-LVEF increased gradually, but decreased at 50 µg/(kg · min). 2D-LVESV and LVMd decreased, but increased at 50 µg/(kg · min). Tmsv-16/12/6 SD%, Tmsv-16/12/6 Dif% decreased, but increased at 50 µg/(kg · min). Eave and Esd increased, but decreased at 40-50 µg/(kg · min). Compared each with the baseline, LV remodeling parameters and VCA were statistically significant. CONCLUSION: IMR patients showed a biphasic response during RT3D-DSE. The decrease of VCA is associated with an improvement of LV remodeling and synchronization when Dob ≤ 40 µg/(kg · min).

[Value of three-dimensional speckle-tracking echocardiography and the novel area strain in detecting wall motion abnormalities of coronary artery diseases].

OBJECTIVE: To investigate the value of three-dimensional speckle tracking echocardiography (3DSTE) and the novel areas train (AS) in detecting wall motion abnormalities caused by coronary artery diseases. METHODS: Conventional and 3D echocardiography was done on forty-six patients suspected with coronary heart diseases, immediately before they received cardiac catheterization. Segmental wall-motion score (WMS) and WMS index (WMSI) were assessed on conventional two-dimensional (2D) images, and strain-derived parameters were analyzed by speckle tracking technique based on 3D images. The accuracy of echocardiographic parameters to assess the regional myocardial ischemia was tested against coronary angiography results using ROC curve analysis and linear regression. RESULTS: Compared with controls, patients with severe coronary stenosis (> or = ;70% coronary stenosis) had lower left ventricular ejection fraction, higher WMSI and lower global strains. ROC curve analysis demonstrated that segmental WMS had relatively poor sensitivity for regional myocardial ischemia and infarction. The segmental strains measured using 3DSTE provided accurate assessment of the regional wall motion abnormalities. The segmental area strain > -25% yielded the best sensitivity and specificity for detecting the ischemic and infracted segments among all four strain parameters. CONCLUSION: 3DSTE provides a swift and accurate assessment of regional myocardial motion. The novel area strain derived by 3DSTE is an accurate and reproducible index for regional wall motion abnormality.

Value of three-dimensional speckle-tracking in detecting left ventricular dysfunction in patients with aortic valvular diseases.

BACKGROUND: The aim of this study was to investigate the value of three-dimensional speckle-tracking echocardiography for the detection of subclinical left ventricular dysfunction in patients with aortic valvular disease (AVD). METHODS: Fifty-nine patients with AVD in New York Heart Association functional class I or II as well as 48 controls were recruited. Patients with AVD were divided further into those with aortic stenosis (AS; n = 34) and those with aortic regurgitation (AR; n = 25). All patients underwent conventional echocardiography and three-dimensional speckle-tracking echocardiography. RESULTS: Analysis of variance showed global longitudinal strain to be compromised in the AR group (-16.9% vs -19.3%, P = .015) and more dramatically decreased in the AS group (-14.3% vs -19.3%, P < .001) compared with healthy controls. Impairment of global circumferential strain was observed in patients with AR (-15.9% vs -18.5%, P = .009) but not in those with AS (-18.2% vs -18.5%, P = .768). Global area strain and global radial strain were decreased in patients with AS and those with AR compared with controls, but the difference between the two subgroups was not significant. CONCLUSIONS: Strains measured by three-dimensional speckle-tracking echocardiography are useful indices of early-stage heart dysfunction caused by AVD. Longitudinal strain is more vulnerable to pressure overload caused by AS, whereas circumferential strain is more sensitive to volume overload due to AR.

Germinable soil seed banks and the restoration potential of abandoned cropland on the Chinese hilly-gullied loess plateau.

Poor vegetation cover is generally considered to be a major factor causing soil erosion on the Loess Plateau in China. It has been argued that tree planting restoration is ineffective, and natural re-vegetation is an alternative ecological solution for restoring abandoned cropland and controlling soil erosion. The aims of this study were to investigate the characteristics of soil seed banks and to assess the natural restoration potential of abandoned cropland in the hilly-gullied Loess Plateau. The soil seed bank was identified by the germination method with the soil samples, which were collected at four sampling times (April, August, and October 2005 and August 2006) from 12 plots abandoned 3-30 years prior to sampling. The seed bank densities of all of the samples in the 0-10 cm soil layer varied from 1,067 ± 225 to 14,967 ± 1,606 seeds m(-2). Fifty-one species (24 annual and 27 perennial species) belonging to 18 families were identified, and 39% of these species belonged to the families Compositae and Gramineae. The pioneer species Artemisia scoparia dominated the seed bank, with an average seed density of 3,722 seeds m(-2), and accounted for 74.4% of the seeds in the bank. The local dominant species (such as Lespedeza davurica, Artemisia gmelinii, Bothriochloa ischaemun and Stipa bungeana) of the later succession stages also existed at densities varying from 17 to 1, 383 seeds m(-2). The combination of soil seed bank characteristics, reproductive traits of the species, the specific landscape conditions indicates that the potential to restoring the abandoned croplands in the hilly-gullied Loess Plateau via natural re-vegetation could be substantial.

[Role of transoesophageal echocardiography on periventricular closure of ventricular septal defects].

OBJECTIVE: To evaluate the clinical value of transoesophageal echocardiography on periventricular closure of ventricular septal defects (VSD). METHODS: Transoesophageal echocardiography (TEE) was performed in 27 patients with VSD diagnosed by the transthoracic echocardiography examination (TTE) and prepared to receive periventricular closure of VSD for the purposes of choosing the appropriate occluder before VSD closure and guiding occluder release during the operation and evaluating the post operative effects of VSD closure 1 week post operation. RESULTS: Three patients with VSD not suitable for periventricular VSD closure according to TEE results were not operated, periventricular closure of VSD were successfully performed in 20 patients and minimal residual shunt was seen in 1 patient post operation, periventricular closure of VSD failed in 4 patients and these patients received surgical closure. TEE at 1 week post successfully with periventricular closure of VSD in the 20 patients showed that there were no residual shunts and the pulmonary artery pressure was significantly lower post operation compared to that of preoperation value. CONCLUSION: TEE is helpful in choosing the suitable patients and occlude size, guiding occlude release and evaluating the post operative results for periventricular closure of ventricular septal defects.

2,2'-Dichloro-1,1'-[(butane-1,4-diyldi-oxy)bis-(nitrilo-methyl-idyne)]dibenzene.

The mol-ecule of the title compound, C(18)H(18)Cl(2)N(2)O(2), lies across a crystallographic inversion centre and adopts an E configuration with respect to the azomethine C=N bond. The imino group is coplanar with the aromatic ring. Within the mol-ecule, the planar units are parallel, but extend in opposite directions from the dimethyl-ene bridge. In the crystal structure, the title compound exhibits a layer packing structure via weak π-π stacking inter-actions [inter-molecular plane-to-plane distances between adjacent aromatic rings are 3.461 (3) Å]. Mol-ecules in each layer are linked by inter-molecular C-H⋯O hydrogen-bonding inter-actions.

2,2'-Dichloro-1,1'-[(propane-1,3-diyldi-oxy)bis-(nitrilo-methyl-idyne)]dibenzene.

The title compound, C(17)H(16)Cl(2)N(2)O(2), assumes a V-shape configuration with a dihedral angle between the two halves of the mol-ecule of 79.60 (4)°. The asymmetric unit comprises one half-mol-ecule with a crystallographic twofold rotation axis passing through the central C atom. There are weak inter-molecular π-π stacking inter-actions between neighbouring benzene rings with inter-molecular plane-to-plane distances of 3.277 (6) and 3.465 (5) Šalong the a and c axes, respectively. In the crystal structure, weak inter-molecular C-H⋯O bonds link each mol-ecule to four others to form an infinite three-dimensional network.