Four-Color SERS Monitoring of Size-dependent Nanoparticle Delivery in the Same Tumor.

The physicochemical properties of nanoparticles (NPs) significantly influence their deposition at the disease site, ultimately impacting the overall therapeutic efficacy; however, precisely assessing the effects of various factors on NP accumulation within a single cell/tumor tissue is challenging due to the lack of appropriate labeling techniques. Surface-enhanced Raman spectroscopy (SERS) tag is a powerful encoding method that has recently been intensively employed for immunodetection of biomarkers. Herein, we introduce a multiplexed SERS tracking approach for systematic investigation of size-dependent accumulation and distribution of NPs within the same tumor. Four-sized (34, 60, 108, and 147 nm) NPs encoded with different SERS "colors" were fabricated, mixed, and incubated with monolayer tumor cells, multicellular tumor spheroids, or injected into mouse models bearing xenograft solid tumors in a single dose. Multicolor SERS detection of the specimens revealed that NP accumulation in tumor cells, tumor spheroids, and solid tumors was in the order of 34 nm > 60 nm > 108 nm > 147 nm, 60 nm > 34 nm > 108 nm > 147 nm, and 34 nm > 147 nm > 108 nm > 60 nm, respectively. Inductively coupled plasma mass spectroscopy determination performed in parallel samples were in alignment with the four-color SERS probing results, demonstrating the effectiveness of this multiplexed evaluation assay. Furthermore, in combination with fluorescence labeling of specific biomolecules, this method can be applied for the colocalization of different NPs in various pathological structures and provide additional information for analysis of the possible mechanisms.

2, 2-dimethylthiazolidine hydrochloride protects against experimental contrast-induced acute kidney injury via inhibition of tubular ferroptosis.

Contrast-induced acute kidney injury (CI-AKI) has become the third leading cause of AKI acquired in hospital, lacking of effective interventions. In the study, we identified the renal beneficial role of 2, 2-dimethylthiazolidine hydrochloride (DMTD), a safer compound which is readily hydrolyzed to cysteamine, in the rodent model of CI-AKI. Our data showed that administration of DMTD attenuated the impaired renal function and tubular injury induced by the contrast agent. Levels of MDA, 4-hydroxynonenal, ferrous iron and morphological signs showed that contrast agent induced ferroptosis, which could be inhibited in the DMTD group. In vitro, DMTD suppressed ferroptosis induced by ioversol in the cultured tubular cells. Treatment of DMTD upregulated glutathione (GSH) and glutathione peroxidase 4 (GPX4). Moreover, we found that DMTD promoted the ubiquitin-mediated proteasomal degradation of Keap1, and thus increased the activity of nuclear factor erythroid 2-related factor 2 (Nrf2). Mechanistically, increase of the ubiquitylation degradation of Keap1 mediates the upregulated effect of DMTD on Nrf2. Consequently, activated Nrf2/Slc7a11 results in the increase of GSH and GPX4, and therefore leads to the inhibition of ferroptosis. Herein, we imply DMTD as a potential therapeutic agent for the treatment of CI-AKI.

Role of body composition and metabolic parameters extracted from baseline 18F-FDG PET/CT in patients with diffuse large B-cell lymphoma.

This study aimed to clarify the clinical and prognostic role of body composition and metabolic parameters extracted from baseline 18F-FDG PET/CT in patients with diffuse large B-cell lymphoma (DLBCL). We retrospectively collected the clinicopathological and 18F-FDG PET/CT parameters of 181 DLBCL patients. The indexes of skeletal muscle, subcutaneous adipose tissue, and visceral adipose tissue were calculated using the area measured at the 3rd lumbar level normalized for height. Additionally, the metabolic activity of corresponding muscle and adipose tissue, and maximum standardized uptake value (SUVmax), metabolic tumor volume (MTV), and total lesion glycolysis (TLG) of all lesions were measured. Survival endpoints included progression-free survival (PFS) and overall survival (OS). We identified 75 (41.4%) patients with low skeletal muscle index (sarcopenia), presenting risk factors including male, high ß2-microglobulin, low BMI, high visceral adipose tissue index, low SUVmax of skeletal muscle, and high SUVmax of visceral adipose tissue. Male, low BMI, low visceral adipose tissue index, and high SUVmax of subcutaneous adipose tissue were risk factors for low subcutaneous adipose tissue index diagnosed in 105 (58.0%) patients. In total, 132 (79.2%) patients represented low visceral adipose tissue index, associated with younger age, B symptoms, and low BMI. Eastern Cooperative Oncology Group (ECOG) status, sarcopenia, and visceral adipose tissue index were found independently predictive of PFS and OS, while ß2-microglobulin was independently predictive of OS. In conclusion, body composition indexes were correlated with both clinical characteristics and 18F-FDG PET/CT metabolic parameters, significantly impacting survival, such that sarcopenia and high visceral adipose tissue index were powerful predictors of poor DLBCL outcomes.

Clinical and prognostic role of 2-[18F]FDG PET/CT and sarcopenia in treatment-naïve patients with T-cell lymphoblastic lymphoma.

T-cell lymphoblastic lymphoma (T-LBL) is a rare and highly aggressive non-Hodgkin lymphoma. This study aimed to explore the role of 2-[18F]FDG PET/CT, sarcopenia, clinical features, and treatment regimens in 49 treatment-naïve patients with T-LBL, and assess their predictive value in the prognosis. Sarcopenia was measured as skeletal muscle index (SMI) at L3 level from the CT component of PET/CT images. All 49 patients (35 males, 14 females; median age, 26 years [range, 3-66 years]) were enrolled in this study, including 36 adult patients and 13 pediatric patients. Lymph nodes, thymus, bone marrow, and pleura were the most common involved sites of T-LBL. The median SUVmax, MTV, and TLG of all lesions in these 49 patients were 12.4 (range, 4.2-40.5), 532.6 (17.4-3518.1), and 2112.2 (53.9-18,699.2), respectively. Eighteen out of 49 patients (36.7%) were diagnosed with sarcopenia. Sarcopenia patients had lower BMI and SUVmax of muscle at L3 level than non-sarcopenia patients (P < 0.05). Univariate Cox regression analysis indicated that higher MTV and TLG and intrathecal therapy (IT) were associated with longer progression-free survival (PFS) and overall survival (OS), while multivariate Cox regression analysis showed that TLG and IT were independent predictors for PFS, and only IT was an independent predictor for OS. In conclusion, low BMI and SUVmax of muscle at L3 level correlated with sarcopenia in T-LBL patients. Higher initial MTV and TLG and receiving IT were associated with better prognosis in T-LBL patients. TLG and IT, but not sarcopenia, were independent prognostic factors.

68Ga-Labeled Cystine Knot Peptide Targeting Integrin αvß6 for Lung Cancer PET Imaging.

Integrin αvß6 has been considered as a promising biomarker for lung cancer, and its expression is often related to poor prognosis. An αvß6-binding cystine knot peptide R01-MG was previously engineered and validated. Here, we developed a positron emission tomography (PET) probe of R01-MG for imaging αvß6-positive lung cancer. Cystine knot peptide R01-MG was synthesized through solid-phase peptide synthesis chemistry and radiolabeled with 68Ga after being conjugated with 1,4,7,10-tetraazacyclododecane-N,N',N″,N‴-tetraacetic acid (DOTA). The stability of 68Ga-DOTA-R01-MG was analyzed in phosphate-buffered saline (PBS) (pH 7.4) and fetal bovine serum (FBS). The cell uptake assay of the probe was evaluated using αvß6-positive (A549 and H1975) and αvß6-negative (H1299) lung cancer cell lines. In addition, small animal PET imaging and biodistribution studies of 68Ga-DOTA-R01-MG were performed in αvß6-positive and αvß6-negative lung cancer models. Our study showed that 68Ga-DOTA-R01-MG exhibited excellent stability in PBS and FBS. Small animal PET imaging and biodistribution data revealed that 68Ga-DOTA-R01-MG displayed rapid and good tumor uptake in animal models with αvß6-positive lung cancer, and the probe was rapidly cleared from the normal tissues, resulting in good tumor-to-normal tissue contrasts. Meanwhile, no obvious tumor uptake of 68Ga-DOTA-R01-MG was observed in animal models with αvß6-negative lung cancer, demonstrating specific binding of the probe to integrin αvß6. In conclusion, 68Ga-DOTA-R01-MG has great potential to be a promising PET tracer for imaging αvß6-positive lung cancer.

Combined legumain- and integrin-targeted nanobubbles for molecular ultrasound imaging of breast cancer.

Molecular ultrasound imaging is a promising strategy for non-invasive and precise cancer diagnosis. Previously reported ultrasound contrast agents (UCAs) are mostly microbubbles or nanobubbles (NBs) larger than 200 nm, leading to less efficient tumor delivery. Here we synthesized NBs with a small size (~49 nm) and modified the NB surface with alanine-alanine-asparagine (NB-A) or arginine-glycine-aspartic acid peptide (NB-R) for concurrent active targeting towards legumain in tumor cells and integrin in tumor neovasculature. In vitro, the NB-A and NB-R presented echogenicity comparable with SonoVue MBs and showed specific binding with tumors cells and endothelial cells, respectively. In vivo, the combined NB-A/NB-R accumulated in tumor tissues selectively and provided ultrasound signals with prolonged duration and that were significantly stronger than non-targeted NBs, single-targeted NBs and SonoVue MBs. Overall, the dual targeted NBs served as efficient UCAs for specific imaging of breast cancer, and hold great potential for general cancer diagnosis/monitoring in the future.

18F-FDG PET/CT and clinicopathological characteristics of neurolymphomatosis in lymphoma patients.

OBJECTIVE: Neurolymphomatosis (NL) is a rare but serious manifestation defined as invasion of peripheral nervous system by malignant lymphocytes. Thus, this study investigated fluorine-18-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) and clinicopathological characteristics of NL in lymphoma patients. SUBJECTS AND METHODS: Clinicopathological and 18F-FDG PET/CT findings and treatment regimens were retrospectively investigated in 20 lymphoma patients with NL, and analyzed their correlation with progression-free survival (PFS) and overall survival (OS). RESULTS: These 20 lymphoma patients (11 males, 9 females; median age, 49 years) included 10 primary and 10 secondary NL patients. Non-Hodgkin's lymphoma (NHL) was noted in 19 patients, B-cell NHL was associated with 18 cases, and diffuse large B-cell lymphoma was the most common. Notably, 18 patients were aggressive lymphoma while 2 were indolent lymphoma. The affected neural structures included nerve roots (n=17), peripheral nerves (n=3), cranial nerves (n=3), and neural plexus (n=2). Fluorine-18-FDG was avid in all cases, and the median maximum standardized uptake value (SUVmax) of neural and all lesions was 12.2 (range, 3.3-25.6) and 15.0 (range, 4.4-34.2), respectively. The median PFS and OS of all patients were 9.3 and 14.3 months. The 12-month OS rate of 18 patients with aggressive lymphoma receiving intrathecal chemotherapy/autologous stem cell transplants (IT chem/ASCT) was significantly higher than who did not (64.8% vs 15.9%). CONCLUSION: The majority of NL occurred in patients with aggressive lymphoma, of which B-cell NHL were the predominant subtypes. Fluorine-18-FDG PET/CT imaging of NL was mainly characterized by intense glucose accumulation alongside peripheral nerves, and IT chem/ASCT was suggested to improve the outcomes of NL.

18F-FDG PET/CT imaging of multiple intrahepatic Epstein-Barr virus-associated smooth muscle tumors in a pediatric patient after heart transplantation.

Epstein-Barr virus-associated smooth muscle tumor (EBV-SMT) is an exceedingly rare neoplastic disease with a predisposition in immune-compromised individuals, especially in patients with prior transplantation, human immunodeficiency virus infection, or congenital immunodeficiency. Here, we present imaging findings of EBV-SMT in multiphasic contrast-enhanced computed tomography (CT) and fluorine-18-fluorodeoxyglucose (18F-FDG) positron emission tomography (PET)/CT in a two-and-a-half-year-old boy with prior heart transplantation.

Colorimetric analysis of extracellular vesicle surface proteins based on controlled growth of Au aptasensors.

Protein profiling of extracellular vesicles (EVs) provides important information in both clinical cancer diagnosis and relevant biological research studies. Although a variety of bioanalytical techniques have been investigated for EV characterization, limitations such as time-consuming operations, the requirement of large sample volume and demand for specialized instruments hinder their practical applications. Here, we report a simple and wash-free homogeneous colorimetric assay for sensitive detection of surface proteins on EVs. Au nanoparticles were modified with thiolated aptamers to fabricate aptasensors and incubated with EVs. Upon addition of a Au growth reagent, the solution color changed from light red to blue in the presence of target proteins and became deep red when the targets were absent. Expression of CD63, epithelial cell adhesion molecules (EpCAM), and mucin1 in EVs derived from two breast cancer cell lines (MCF-7 and MDA-MB-231) were compared, showing results consistent with western blotting results. The colorimetric assay achieves a limit of detection (LOD) down to 0.7 ng µL-1 against MCF-7 EVs based on the assessment of EpCAM expression, suggesting its potential to be applied in clinical breast cancer diagnosis.

Skipping of an exon with a nonsense mutation in the DMD gene is induced by the conversion of a splicing enhancer to a splicing silencer.

Modulation of dystrophin pre-mRNA splicing is an attractive strategy to ameliorate the severe phenotype of Duchenne muscular dystrophy (DMD), although this requires a better understanding of the mechanism of splicing regulation. Aberrant splicing caused by gene mutations provides a good model to study splicing regulatory cis-elements and binding proteins. In this study, we identified skipping of in-frame exon 25 induced by a nonsense mutation (NM_004006.2:c.3340A > T;p.Lys1114*) in the DMD gene. Site-directed mutagenesis study in minigenes suggested that c.3340A > T converts an exonic splicing enhancer sequence (ESE) to a silencer element (ESS). Indeed, RNA pull-down and functional study provided evidence that c.3340A > T abolishes the binding of the splicing enhancer protein Tra2ß and promotes interactions with the repressor proteins hnRNP A1, hnRNP A2, and hnRNP H. By carefully analyzing the sequence motif encompassing the mutation site, we concluded that the skipping of exon 25 was due to disruption of a Tra2ß-dependent ESE and the creation of a new ESS associated with hnRNP A1 and hnRNP A2, which in turn increased the recruitment of hnRNP H to a nearby binding site. Finally, we demonstrated that c.3340A > T impairs the splicing of upstream intron 24 in a splicing minigene assay. In addition, we showed that the correct splicing of exon 25 is finely regulated by multiple splicing regulators that function in opposite directions by binding to closely located ESE and ESS. Our results clarify the detailed molecular mechanism of exon skipping induced by the nonsense mutation c.3340A > T and also provide information on exon 25 splicing.

Legumain, an asparaginyl endopeptidase, mediates the effect of M2 macrophages on attenuating renal interstitial fibrosis in obstructive nephropathy.

Two distinct macrophage phenotypes contribute to kidney injury and repair during the progression of renal interstitial fibrosis; proinflammatory (M1) and antiinflammatory (M2) macrophages. Legumain, an asparaginyl endopeptidase of the cysteine protease family, is overexpressed in macrophages in some pathological conditions. However, the macrophage subtype and function of macrophage-derived legumain remains unclear. To resolve this we tested whether M2 macrophages contribute to the accumulation of legumain in the unilateral ureteral obstruction model. Legumain-null mice exhibited more severe fibrotic lesions after obstruction compared with wild-type control. In vitro, IL4-stimulated M2 polarization led to the overexpression and secretion of legumain. The levels of fibronectin and collagen I/III, major components of the extracellular matrix, were reduced in the conditioned medium of TGF-ß1-stimulated tubular epithelial cells or fibroblasts after treatment with legumain or conditioned medium from IL4-stimulated macrophages. Administration of the legumain inhibitor RR-11a exacerbated fibrotic lesions following obstruction. Therapeutically, adoptive transfer of legumain-overexpressing macrophages or IL4-stimulated macrophages ameliorated the deposition of collagen and fibronectin induced by ureteral obstruction, either in the wild-type mice or in lgmn-/- mice. Thus, M2 macrophages overexpress and secret legumain and legumain mediates the anti-fibrotic effect of M2 macrophages in obstructive nephropathy.

A resolved discrepancy between multiplex PCR and multiplex ligation-dependent probe amplification by targeted next-generation sequencing discloses a novel partial exonic deletion in the Duchenne muscular dystrophy gene.

BACKGROUND: The genetic diagnosis of Duchenne muscular dystrophy (DMD) has been complicated by the large size of the gene and its heterogeneous mutational spectrum. Multiplex PCR and multiplex ligation-dependent probe amplification (MLPA) are two well-established mutation screening methods. Here, we applied targeted next-generation sequencing (NGS) to clarify discrepant results between multiplex PCR and MLPA in a Chinese patient with DMD. METHODS: MLPA was performed to confirm multiplex PCR results obtained previously. Targeted NGS was then used to analyze the full-length DMD gene including introns. RESULTS: Multiplex PCR had previously identified an apparent deletion of exon 43 in the patient with DMD, but current MLPA indicated that exon 43 was present. Targeted NGS to clarify the genetic diagnosis identified a novel mutation, c.6241_c.6290 + 1109del1159insAC, which caused partial deletion of exon 43. This mutation removed the annealing sequence of the exon 43 reverse primer in multiplex PCR but had no influence on the hybridization site of the MLPA probe. Therefore, the discrepancy between the two methods was caused by partial exonic deletion that escaped MLPA detection. CONCLUSION: Targeted NGS disclosed a novel partial exonic deletion in the DMD gene as the cause of discrepancy between multiplex PCR and MLPA. Targeted NGS could be used to provide a more accurate genetic diagnosis of DMD, particularly in cases of partial exonic deletions, which will be of benefit in patient management and the identification of disease carriers.

Polymeric Nanomedicine for Combined Gene/Chemotherapy Elicits Enhanced Tumor Suppression.

Combination treatment through simultaneous delivery of DNA and anticancer drugs with nanoparticles has been demonstrated to be an elegant and efficient approach for cancer therapy. Herein, we employed a combination therapy for eliminating both the tumor cells and intratumoral neovascular network based on the nanoplatform we designed. Pigment epithelium-derived factor (PEDF) gene, a powerful antiangiogenic agent, and the clinically widely used chemotherapy agent paclitaxel (PTX) were simultaneously encapsulated in the same nanoparticle by a modified double-emulsion solvent evaporation method. The dual-drug-loaded nanoparticles (D/P-NPs) exhibited a uniform spherical morphology and released PTX and PEDF gene in a sustained manner. D/P-NPs showed an enhanced antitumor effect on C26 and A549 cells and a stronger inhibitory activity on proliferation of HUVECs. Moreover, D/P-NPs could dramatically elevate the PEDF expression levels in both C26 and A549 cells in comparison with PEDF gene loaded nanoparticles and significantly promote the cellular uptake of PTX. Additionally, microtubules were stabilized and G2/M phase arrest along with a higher subG1 cell population was induced by D/P-NPs in contrast to PTX or PTX loaded nanoparticles. Besides, D/P-NPs showed sustained release of PTX and PEDF gene in tumors as well as long-term gene expression. A significantly improved anticancer effect was also demonstrated in a C26 subcutaneous tumor model using this combinational therapy. D/P-NPs could sharply reduce the microvessel density and significantly promoted tumor cell apoptosis in vivo. More importantly, the in vivo distribution, serological and biochemical analysis, and H&E staining revealed that D/P-NPs had no obvious toxicity. Our study suggested that this novel polymeric nanomedicine had great potential for improving the therapeutic efficacy of combined gene/chemotherapy of cancer.

Overexpression of Oct4 suppresses the metastatic potential of breast cancer cells via Rnd1 downregulation.

Although Oct4 is known as a critical transcription factor involved in maintaining "stemness", its role in tumor metastasis is still controversial. Herein, we overexpressed and silenced Oct4 expression in two breast cancer cell lines, MDA-MB-231 and 4T1, separately. Our data showed that ectopic overexpression of Oct4 suppressed cell migration and invasion in vitro and the formation of metastatic lung nodules in vivo. Conversely, Oct4 downregulation increased the metastatic potential of breast cancer cells both in vitro and in vivo. Furthermore, we identified Rnd1 as the downstream target of Oct4 by ribonucleic acid sequencing (RNA-seq) analysis, which was significantly downregulated upon Oct4 overexpression. Chromatin immunoprecipitation assays revealed the binding of Oct4 to the promoter region of Rnd1 by ectopic overexpression of Oct4. Dual luciferase assays indicated that Oct4 overexpression suppressed transcriptional activity of the Rnd1 promoter. Moreover, overexpression of Rnd1 partially rescued the inhibitory effects of Oct4 on the migration and invasion of breast cancer cells. Overexpression of Rnd1 counteracted the influence of Oct4 on the formation of cell adhesion and lamellipodia, which implied a potential underlying mechanism involving Rnd1. In addition, we also found that overexpression of Oct4 led to an elevation of E-cadherin expression, even in 4T1 cells that possess a relatively high basal level of E-cadherin. Rnd1 overexpression impaired the promoting effects of Oct4 on E-cadherin expression in MDA-MB-231 cells. These results suggest that Oct4 affects the metastatic potential of breast cancer cells through Rnd1-mediated effects that influence cell motility and E-cadherin expression.

Chronic bronchitis leads to accelerated hyperinflation in COPD patients during exercise.

BACKGROUND AND OBJECTIVE: It is not known whether patients with chronic obstructive pulmonary disease (COPD) have a different exercise capacity with (CB(+) ) or without accompanying chronic bronchitis (CB(-) ). METHODS: We conducted spirometry, a 6-min walk distance test and cardiopulmonary exercise test in 50 age-matched healthy control subjects, 45 COPD patients without CB (CB(-) ) and 37 COPD patients with CB (CB(+) ). A multiple regression model was established to identify factors independently associated with peak oxygen consumption ( V ˙ O 2 ). RESULTS: Patients with and without CB had similar forced expiratory volume in 1 s (FEV1 ). CB(+) patients had a lower V ˙ O 2 . CB(+) and CB(-) participants had similar increases in tidal volume at peak exercise; however, CB(+) patients had an increased respiratory rate (RR). These patients reached the peak value for ratio of end-expiratory lung volume to total lung capacity (TLC) at a lower work load. A stepwise multiple linear regression analysis identified chronic bronchitis, FEV1 , diffusing capacity for carbon monoxide, the ratio of residual volume to TLC and serum tumour necrosis factor-α as independent predictors of peak V ˙ O 2 . CONCLUSIONS: CB significantly lowers exercise capacity in COPD patients because of dynamic hyperinflation during exercise. The accelerated dynamic hyperinflation may contribute to increased airway and systemic inflammation in COPD patients.

[Clinical value of cardiopulmonary exercise testing derived oxygen uptake efficiency parameters in patients with end-stage chronic heart failure].

OBJECTIVE: To assess the cardiopulmonary exercise testing (CPET) derived performance of oxygen uptake and ventilation efficiency parameters, including oxygen uptake efficiency plateau (OUEP) , oxygen uptake efficiency slope (OUES), V·E/V·CO2 slope and lowest V·E/V·CO2, in patients with end-stage chronic heart failure (CHF) and evaluate their clinical value on monitoring cardiac function and hemodynamic status. METHODS: A total of 26 end-stage CHF patients considered for heart transplantation were enrolled in this study. CPET, echocardiography and invasive hemodynamic examinations with Swan-Ganz flowing balloon catheter were performed. Correlation analysis was made between oxygen uptake and ventilation efficiency parameters from CPET and echocardiographic and hemodynamic parameters. RESULTS: OUEP and OUES showed good correlation with peak oxygen consumption (peak V·O2) (r = 0.535, P < 0.01;r = 0.840, P < 0.001). In end-stage CHF patients, the slope of OUEP with respect to peak V·O2 is about 32, but the slope of OUES with respect to peak V·O2 is only about 2. The difference was 16 times. The change of OUEP was more sensitive and significant than those of OUES and peak V·O2 (P < 0.05). OUEP, peak V·O2 (%pred), V·E/V·CO2 slope and lowest V·E/V·CO2 were all correlated well with non-invasive hemodynamic parameters peak cardiac output (r = 0.535, P < 0.01; r = 0.652, P < 0.001; r = -0.640, P < 0.001; r = -0.606, P = 0.001 respectively) and peak cardiac index (r = 0.556, P < 0.01;r = 0.772, P < 0.001; r = -0.641, P < 0.001; r = -0.620, P < 0.001 respectively) derived from CPET, but not correlated with invasive hemodynamic parameters cardiac output and cardiac index at rest (P > 0.05). Both peak V·O2 (%pred) and V·E/V·CO2 slope were significantly correlated with invasive hemodynamic parameters systolic pulmonary arterial pressure (r = -0.424, P < 0.05; r = 0.509, P < 0.01) and mean pulmonary arterial pressure (r = -0.479, P < 0.05; r = 0.405, P < 0.05). Peak V·O2 (%pred) was also significantly correlated with pulmonary capillary wedge pressure (r = -0.415, P < 0.05), and V·E/V·CO2 slope was significantly correlated with pulmonary vascular resistance (r = 0.429, P < 0.05). CONCLUSIONS: The oxygen uptake and ventilation efficiency parameters derived from CPET, including peak V·O2, OUEP, lowest V·E/V·CO2 and V·E/V·CO2 slope etc, are objectively monitoring and evaluating cardiac function and hemodynamic status. And they are useful for optimizing clinical management of patients with end-stage CHF.

Influence of bushenhuoxue on podocytes of focal segmental glomerulosclerosis mice.

OBJECTIVE: To observe the effects and mechanisms of Bushenhuoxue on desmin and nephrin expression in mice podocytes, and to investigate its effects on wt1 expression in Wilms' tumor. METHODS: Adriamycin (ADR) was used to induce focal segmental glomerulous sclerosis (FSGS) in mice. Bushenhuoxue was used to treat FSGS for 6 weeks. We measured body mass and right renal mass, and determined serum albumin (ALB) levels, protein content in urine, and urinary protein and albumin creatinine ratio (UACR). Changes in renal tissue morphology were evaluated by microscopy. wt1 and nephrin expression in podocytes were detected using immunofluorescence. Expression levels of desmin, wt1 and nephrin mRNAs in renal tissue were determined using reverse transcription polymerase chain reaction assays. RESULTS: Protein levels in urine and UACR were significantly increased in FSGS model mice compared with Bushenhuoxue-treated and control mice. Body mass and ALB levels were decreased in FSGS mice compared with control and Bushenhuoxue-treated mice. Expression of the wt1 protein was observed in control mice. Compared with controls, wt1 expression levels were reduced in Bushenhuoxue-treated mice, and to a greater extent in FSGS mice. Nephrin protein expression was widespread in FSGS mice, and significantly reduced in control and Bushenhuoxue mice. Expression levels of wt1 and nephrin mRNAs in FSGS mice were lower compared with those in control and Bushenhuoxue-treated mice. Desmin mRNA levels in FSGS mice were reduced compared with those in control and Bushenhuoxue-treated mice. CONCLUSION: Bushenhuoxue ameliorated albuminuria in FSGS mice; this was possibly related to the up-regulation of wt1 and nephrin, and down-regulation of desmin.

[Parameters of oxygen uptake and carbon dioxide output ventilatory efficiency during exercise are index of circulatory function in normal subjects].

OBJECTIVE: To observe oxygen uptake efficiency plateau (OUEP, i.e.highest V˙O2/V˙E) and carbon dioxide output efficiency (lowest V˙E/V˙CO2) parameter changes during exercise in normal subjects. METHODS: Five healthy volunteers performed the symptom limited maximal cardiopulmonary exercise test (CPET) at Harbor-UCLA Medical Center. V˙O2/V˙E and V˙E/V˙CO2 were determined by both arterial and central venous catheters. After blood gas analysis of arterial and venous sampling at the last 30 seconds of every exercise stage and every minute of incremental loading, the continuous parameter changes of hemodynamics, pulmonary ventilation were monitored and oxygen uptake ventilatory efficiency (V˙O2/V˙E and V˙E/V˙CO2) was calculated. RESULTS: During CPET, as the loading gradually increased, cardiac output, heart rate, mixed venous oxygen saturation, arteriovenous oxygen difference, minute ventilation, minute alveolar ventilation, tidal volume, alveolar ventilation and pulmonary ventilation perfusion ratio increased near-linearly (P < 0.05-0.01, vs.resting); arterial oxygen concentration maintained at a high level without significant change (P > 0.05); stroke volume, respiratory rate, arterial partial pressure of carbon dioxide, arterial blood hydrogen ion concentration and dead space ventilation ratio significantly changed none-linearly (compare resting state P < 0.05-0.01).OUE during exercise increased from 30.9 ± 3.3 at resting state to the highest plateau 46.0 ± 4.7 (P < 0.05 vs.resting state), then, declined gradually after anaerobic threshold (P < 0.05-0.01, vs.OUEP) and reached 36.6 ± 4.4 at peak exercise. The V˙E/V˙CO2 during exercise decreased from the resting state (39.2 ± 6.5) to the minimum value (24.2 ± 2.4) after AT for a few minutes (P > 0.05 vs.earlier stage), then gradually increased after the ventilatory compensation point (P < 0.05 vs.earlier stage) and reached to 25.9 ± 2.7 at peak exercise. CONCLUSIONS: Cardiac and lung function as well as metabolism change during CPET is synchronous.In the absence of pulmonary limit, appearing before and after anaerobic threshold, OUEP and lowest V˙E/V˙CO2 could be used as reliable parameters representing the circulatory function.

11C-CFT PET brain imaging in Parkinson's disease using a total-body PET/CT scanner.

PURPOSE: This study aimed to evaluate the feasibility of 11C-CFT PET brain imaging in Parkinson's Disease using a total-body PET/CT scanner and explore the optimal scan duration to guide the clinical practice. METHODS: Thirty-two patients with Parkinson's disease (PD) performing 11C-CFT PET/CT brain imaging using a total-body PET/CT scanner were retrospectively enrolled. The PET data acquired over a period of 900 s were reconstructed into groups of different durations: 900-s, 720-s, 600-s, 480-s, 300-s, 180-s, 120-s, 60-s, and 30-s (G900 to G30). The subjective image quality analysis was performed using 5-point scales. Semi-quantitative measurements were analyzed by SUVmean and dopamine transporter (DAT) binding of key brain regions implicated in PD, including the caudate nucleus and putamen. The full-time images (G900) were served as reference. RESULTS: The overall G900, G720, and G600 image quality scores were 5.0 ± 0.0, 5.0 ± 0.0, and 4.9 ± 0.3 points, respectively, and there was no significant difference among these groups (P > 0.05). A significant decrease in these scores at durations shorter than 600 s was observed when compared to G900 images (P < 0.05). However, all G300 image quality was clinically acceptable (≥ 3 points). As the scan duration reduced, the SUVmean and DAT binding of caudate nucleus and putamen decreased progressively, while there were no statistically significant variations in the SUVmean of the background among the different groups. Moreover, the changes in the lesion DAT binding (ΔDAT-binding) between the full-time reference G900 image and other reconstructed group G720 to G30 images generally increased along with the reduced scan time. CONCLUSION: Sufficient image quality and lesion conspicuity could be achieved at 600-s scan duration for 11C-CFT PET brain imaging in PD assessment using a total-body PET/CT scanner, while the image quality of G300 was acceptable to meet clinical diagnosis, contributing to improve patient compliance and throughput of PET brain imaging.

Implications of Sarcopenia and Glucometabolism Parameters of Muscle Derived From Baseline and End-of-Treatment 18F-FDG PET/CT in Diffuse Large B-Cell Lymphoma.

OBJECTIVE: We previously found that the incidence of sarcopenia increased with declining glucose metabolism of muscle in patients with treatment-naïve diffuse large B-cell lymphoma (DLBCL). This study aimed to investigate the relationship between sarcopenia and muscle glucometabolism using 18F-FDG PET/CT at baseline and end-of-treatment, analyze the changes in these parameters through treatment, and assess their prognostic values. MATERIALS AND METHODS: The records of 103 patients with DLBCL (median 54 years [range, 21-76]; male:female, 50:53) were retrospectively reviewed. Skeletal muscle area at the third lumbar vertebral (L3) level was measured, and skeletal muscle index (SMI) was calculated to determine sarcopenia, defined as SMI < 44.77 cm²/m² and < 32.50 cm²/m² for male and female, respectively. Glucometabolic parameters of the psoas major muscle, including maximum standardized uptake value (SUVmax) and mean standardized uptake value (SUVmean), were measured at L3 as well. Their changes across treatment were also calculated as ΔSMI, ΔSUVmax, and ΔSUVmean; Δbody mass index was also calculated. Associations between SMI and the metabolic parameters were analyzed, and their associations with progression-free survival (PFS) and overall survival (OS) were identified. RESULTS: The incidence of sarcopenia was 29.1% and 36.9% before and after treatment, respectively. SMI (P = 0.004) was lower, and sarcopenia was more frequent (P = 0.011) at end-of-treatment than at baseline. The SUVmax and SUVmean of muscle were lower (P < 0.001) in sarcopenia than in non-sarcopenia at both baseline and end-of-treatment. ΔSMI was positively correlated with ΔSUVmax of muscle (P = 0.022). Multivariable Cox regression analysis showed that sarcopenia at end-of-treatment was independently negatively associated with PFS (adjusted hazard ratio [95% confidence interval], 2.469 [1.022-5.965]), while sarcopenia at baseline was independently negatively associated with OS (5.051 [1.453-17.562]). CONCLUSION: Sarcopenic patients had lower muscle glucometabolism, and the muscular and metabolic changes across treatment were positively correlated. Sarcopenia at baseline and end-of-treatment was negatively associated with the prognosis of DLBCL.