Integrated spatially resolved metabolomics and network toxicology to investigate the hepatotoxicity mechanisms of component D of Polygonum multiflorum Thunb.

ETHNOPHARMACOLOGICAL RELEVANCE: The liver toxicity of Reynoutria multiflora (Thunb.) Moldenke. (Polygonaceae) (Polygonum multiflorum Thunb, PM) has always attracted much attention, but the related toxicity materials and mechanisms have not been elucidated due to multi-component and multi-target characteristics. In previous hepatotoxicity screening, different components of PM were first evaluated and the hepatotoxicity of component D [95% ethanol (EtOH) elution] in a 70% EtOH extract of PM (PM-D) showed the highest hepatotoxicity. Furthermore, the main components of PM-D were identified and their hepatotoxicity was evaluated based on a zebrafish embryo model. However, the hepatotoxicity mechanism of PM-D is unknown. AIM OF THE STUDY: This work is to explore the hepatotoxicity mechanisms of PM-D by integrating network toxicology and spatially resolved metabolomics strategy. MATERIALS AND METHODS: A hepatotoxicity interaction network of PM-D was constructed based on toxicity target prediction for eight key toxic ingredients and a hepatotoxicity target collection. Then the key signaling pathways were enriched, and molecular docking verification was implemented to evaluate the ability of toxic ingredients to bind to the core targets. The pathological changes of liver tissues and serum biochemical assays of mice were used to evaluate the liver injury effect of mice with oral administration of PM-D. Furthermore, spatially resolved metabolomics was used to visualize significant differences in metabolic profiles in mice after drug administration, to screen hepatotoxicity-related biomarkers and analyze metabolic pathways. RESULTS: The contents of four key toxic compounds in PM-D were detected. Network toxicology identified 30 potential targets of liver toxicity of PM-D. GO and KEGG enrichment analyses indicated that the hepatotoxicity of PM-D involved multiple biological activities, including cellular response to endogenous stimulus, organonitrogen compound metabolic process, regulation of the apoptotic process, regulation of kinase, regulation of reactive oxygen species metabolic process and signaling pathways including PI3K-Akt, AMPK, MAPK, mTOR, Ras and HIF-1. The molecular docking confirmed the high binding activity of 8 key toxic ingredients with 10 core targets, including mTOR, PIK3CA, AKT1, and EGFR. The high distribution of metabolites of PM-D in the liver of administrated mice was recognized by mass spectrometry imaging. Spatially resolved metabolomics results revealed significant changes in metabolic profiles after PM-D administration, and metabolites such as taurine, taurocholic acid, adenosine, and acyl-carnitines were associated with PM-D-induced liver injury. Enrichment analyses of metabolic pathways revealed tht linolenic acid and linoleic acid metabolism, carnitine synthesis, oxidation of branched-chain fatty acids, and six other metabolic pathways were significantly changed. Comprehensive analysis revealed that the hepatotoxicity caused by PM-D was closely related to cholestasis, mitochondrial damage, oxidative stress and energy metabolism, and lipid metabolism disorders. CONCLUSIONS: In this study, the hepatotoxicity mechanisms of PM-D were comprehensively identified through an integrated spatially resolved metabolomics and network toxicology strategy, providing a theoretical foundation for the toxicity mechanisms of PM and its safe clinical application.

Enzymatically functionalized RGD-gelatin scaffolds that recruit host mesenchymal stem cells in vivo and promote bone regeneration.

Critical-size bone defects are imposing a substantial biomedical burden. Despite being long regarded as a potential approach to mitigate this burden or an alternative to bone grafts, bone tissue engineering (BTE) has virtually not proceeded to widespread clinical practices. In the BTE field, it is highly required to find a facile method to prepare active scaffolds with tailored biological functions. Here, we immobilized cell adhesive RGD motifs onto gelatin sponge (GS) scaffolds through enzymatic linking. On the basis of the resulting RGD-functionalized GS (RGD/GS) scaffolds, we developed a new and convenient strategy for bone defect repair, in which the scaffolds were first used to recruit mesenchymal stem cells (MSCs) from skeletal muscle, immediately followed by their engraftment into bone defect. We demonstrated significantly enhanced host cells homing into RGD/GS scaffolds as a result of specific RGD-integrin interactions, and the recruited host cells showed a strong osteogenic differentiation potential. After ectopic implantation of cell-laden RGD/GS scaffolds into critical-size mouse bone defects, marked bone tissue regeneration occurred. The presented strategy not only provides an agile route for the preparation of bioactive scaffolds and the construction of osteoinductive bone-graft substitutes, but also avoids or minimizes the complicated and laborious cell isolation, in vitro expansion and cell seeding procedures used in the conventional BTE.

Anti-anaphylactic potential of benzoylpaeoniflorin through inhibiting HDC and MAPKs from Paeonia lactiflora.

Guided by cell-based anti-anaphylactic assay, eighteen cage-like monoterpenoid glycosides (1-18) were obtained from the bioactive fraction of P. lactiflora extract. Among these, compounds 1, 5, 6, 11, 12, 15, and 17 significantly reduced the release rate of ß-HEX and HIS without or with less cytotoxicity. Furthermore, the most potent inhibitor benzoylpaeoniflorin (5) was selected as the prioritized compound for the study of action of mechanism, and its anti-anaphylactic activity was medicated by dual-inhibiting HDC and MAPK signal pathway. Moreover, molecular docking simulation explained that benzoylpaeoniflorin (5) blocked the conversion of L-histidine to HIS by occupying the HDC active site. Finally, in vivo on PCA using BALB/c mice, benzoylpaeoniflorin (5) suppressed the IgE-mediated PCA reaction in antigen-challenged mice. These findings indicated that cage-like monoterpenoid glycosides, especially benzoylpaeoniflorin (5), mainly contribute to the anti-anaphylactic activity of P. lactiflora by dual-inhibiting HDC and MAPK signal pathway. Therefore, benzoylpaeoniflorin (5) may be considered as a novel drug candidate for the treatment of anaphylactic diseases.

Rapid Hemostasis Resulting from the Synergism of Self-Assembling Short Peptide and O-Carboxymethyl Chitosan.

The development of novel hemostatic agents with distinct modes of action from traditional ones remains a formidable challenge. Self-assembling peptide hydrogels have emerged as a new hemostatic material, not only because of their inherent biocompatibility and biodegradability but also their designability. Especially, rational molecular design can make peptides and their hydrogelation responsive to biological cues. In this study, we demonstrated that transglutaminase-catalyzed reactions not only occurred among designed short peptide I3QGK molecules but also between the peptide and a natural polysaccharide O-carboxymethyl chitosan. Because Factor XIII in the blood can rapidly convert into activated transglutaminase (Factor XIIIa) upon bleeding, these enzymatic reactions, together with the electrostatic attraction between the two hemostatic agents, induced a strong synergetic effect in promoting hydrogelation, blood coagulation, and platelet adhesion, eventually leading to rapid hemostasis. The study presents a promising strategy for developing alternative hemostatic materials and methods.

5-HMF induces anaphylactoid reactions in vivo and in vitro.

AIM: Excessive exposure to 5-hydroxymethylfurfural (5-HMF), which is a common impurity in various sugar-containing products, induces serious side effects. Our previous study revealed that 5-HMF exerted immune sensitizing potential when injected into rodents. In this study, we explored 5-HMF mediated anaphylactoid reactions and its underlying molecular mechanisms. METHODS: We investigated anaphylactoid reactions in Brown Norway (BN) rats and Institute of Cancer Research (ICR) mice to identify 5-HMF mediated in vivo anaphylactoid reactions. RBL-2H3 and P815 cell degranulation models were also established, and degranulation, enzyme-linked immunosorbent, filamentous actin (F-actin) microfilament staining, and western blot assays were performed in these cells. RESULTS: We showed that 5-HMF induced anaphylactoid reactions by increasing blood vessel permeability in mice, and significantly elevating histamine (His) and glutathione peroxidase-1 (Gpx-1) levels in rat serum. Moreover, after incubation with 5-HMF, ß-hexosaminidase (ß-Hex), His, IL-4 and IL-6 levels were all significantly increased, thereby inducing cellular degranulation in RBL-2H3 and P815 cells. Finally, 5-HMF also upregulated Lyn, Syk, p38 and JNK protein phosphorylation levels. CONCLUSIONS: Our findings suggest that 5-HMF induces anaphylactoid reactions both in vivo and in vitro, therefore 5-HMF limits in sugar-containing products should receive more regulatory attention.

EBV-Associated T-Cell Lymphoproliferative Disorders Demonstrated on FDG PET/CT in a Patient With Hemophagocytic Lymphohistiocytosis.

FDG PET/CT was performed in a 20-year-old woman to find the underlying cause of hemophagocytic lymphohistiocytosis. The images revealed hypermetabolic activity in multiple lymph nodes and in the spleen. Lymphoma was suspected. However, the pathology of bone marrow, lymph nodes, and the spleen demonstrated chronic active Epstein-Barr virus-associated T-cell lymphoproliferative disorders.

Transglutaminase-Triggered Gelation and Functionalization of Designed Self-Assembling Peptides for Guiding Cell Migration.

Due to the structural and compositional similarity to the extracellular matrix (ECM), self-assembling peptide hydrogels (SAPHs) have shown potential as three-dimensional (3D) scaffolds in regenerative medicine. However, how to fabricate SAPHs with controlled bioactivity for specific applications still remains greatly challenging. In this study, we develop a viable strategy to prepare bioactive SAPHs based on designed short amphiphilic peptides and cell-adhesive motifs, through the combination of enzymatic functionalization and gelation methods. Transglutaminase (TGase) can successfully catalyze the tethering of bioactive features onto the peptide nanofiber surface, and the TGase-meditated conjugation is found to rapidly trigger peptide gelation. Confocal microscopy indicates a uniform ligand distribution within the fibrous hydrogel, and mass spectrometry measurements reveal the involved enzymatic reactions and the gelation mechanism. Furthermore, the preosteoblast cells introduced on the bioactive gel surface are found to be able to migrate into the gel to form 3D distribution and further undergo differentiation, primarily directed by the cell-adhesive motifs within the gel. Such a TGase-based strategy for reconstituting bioactive hydrogels can be readily applied to any other self-assembling peptides and active motifs given rational molecular design, thus showing broad-spectrum application potentials in biological and biomedical fields.

[Effect of Compound Tongfu Granule on Intestinal Permeability in Elderly Sepsis Patients].

OBJECTIVE: To explore the effect of Compound Tongtu Granule (CTG) on intestinal permeability in elderly sepsis patients. METHODS: Eighty elderly sepsis patients were randomly assigned to the experimental group and the control group by randomized double blinded method, 40 in each group. On the basis of conventional antiseptic treatment program, patients in the experimental group took CTG, while those in the control group took placebos. The dosage for CTG or placebos was 14.3 g each package, one package each time, twice daily for 14 successive days. Patients' abdominal symptoms and signs, levels of serum inflammatory factors (high-sensitivity C-reactive protein and procalcitonin), levels of plasma endotoxin, and the intestinal permeability (IP, represented by urinary lactulose/mannitol excretion rate) were compared between the two groups before and after treatment. RESULTS: After 14-day treatment, patients in the experimental group had improved abdominal symptoms, increased frequency of defecation, significantly decreased levels of plasma endotoxin and IP, when compared with the control group (P < 0.05). CONCLUSION: CTG could improve the intestinal barrier function in elderly sepsis patients.

The role of cholescintigraphy in demonstrating delayed post prandial gallbladder motility in cirrhotic patients.

OBJECTIVE: The purpose of this study was to investigate the postprandial gallbladder motility of cirrhotic patients using cholescintigraphy by technetium-99m-ethylhydroiminodiacetic acid ((99m)Tc-EHIDA). SUBJECTS AND METHODS: Sixty two cirrhotic patients (Child-Pugh A: 28 patients; Child-Pugh B: 21 and Child-Pugh C: 13 patients) and 24 normal subjects were included in this study. All patients underwent cholescintigraphy. Mean gallbladder ejection fraction (GBEF) and mean ejection period (EP) were acquired by the region of interest method. In order to evaluate gallbladder contractility of cirrhotic patients, their mean GBEF and EP were compared with the same tests of normal subjects using an independent sample t test. RESULTS: The mean GBEF of cirrhotic patients was lower and their mean ER was longer than that of normal subjects. The means of GBEF and EP of cirrhotic patients were different among different Child-Pugh grade groups. All these differences showed statistical significance. CONCLUSIONS: This study suggests that post prandial cholescintigraphy by (99m)Tc EHIDA demonstrated slower gallbladder motility in cirrhotic patients. The ejection period of cholescintigraphy by (99m)Tc-EHIDA can be used as an index of abnormal gallbladder motility.

Value of dual-phase (99m)Tc-sestamibi scintigraphy with neck and thoracic SPECT/CT in secondary hyperparathyroidism.

OBJECTIVE: Surgical intervention in the form of parathyroidectomy is generally considered only for severe secondary hyperparathyroidism (sHPT). However, correct location of the parathyroid glands before parathyroidectomy is a challenge. The purpose of this study was to compare the diagnostic value of early and delayed phase (99m)Tc-sestamibi SPECT/CT in the detection of parathyroid tissue to guide operative treatment of patients with sHPT. SUBJECTS AND METHODS: Eighty patients with sHPT who were undergoing hemodialysis were evaluated preoperatively with dual-phase (99m)Tc-sestamibi SPECT/CT parathyroid scintigraphy to locate parathyroid tissue before parathyroidectomy. The scintigraphic results were classified as positive or negative. The accuracy of (99m)Tc sestamibi early and delayed phase SPECT/CT scintigraphy was determined. RESULTS: Early phase (99m)Tc-sestamibi SPECT/CT depicted 3.57 parathyroid glands (PTGs) and delayed phase (99m)Tc-sestamibi SPECT/CT depicted 3.55 PTGs per study. The specificity of both early and delayed phase (99m)Tc-sestamibi SPECT/CT in detecting PTGs was 100%. The (99m)Tc-sestamibi SPECT/CT images of 7 of 80 patients showed positive findings in the delayed phase and negative findings in the early phase. The (99m)Tc-sestamibi SPECT/CT images of 6 of 80 patients showed positive findings in the early phase and negative findings in the delayed phase. CONCLUSION: The results of our study indicate that both early and delayed phase (99m)Tc-sestamibi SPECT/CT should be performed in the preoperative evaluation of hemodialysis patients with sHPT due to chronic kidney disease. Performance of both early and delayed phase (99m)Tc-sestamibi SPECT/CT did not increase the radiation dose compared with the use of only the early or the delayed phase.

Diagnostic performance of 18F-FDG PET/CT in patients with fever of unknown origin: a meta-analysis.

PURPOSE: The aim of this study was to systematically review and perform a meta-analysis of published data on the diagnostic value of F-fluorodeoxyglucose (F-FDG) PET/CT in the diagnosis of patients with fever of unknown origin (FUO). MATERIALS AND METHODS: A comprehensive computer literature search of studies published through 31 March 2012 on the use of F-FDG PET/CT in patients with FUO was performed in PubMed/MEDLINE, Embase, and Scopus databases. The pooled sensitivity of F-FDG PET/CT in patients with FUO on a per-patient-based analysis was calculated. The area under the receiver-operating characteristic curve was calculated to measure the accuracy of F-FDG PET/CT in the diagnosis of patients with FUO. RESULTS: Fifteen studies comprising 595 patients with FUO were included in this meta-analysis. The pooled sensitivity of F-FDG PET/CT in detecting the cause of FUO was 85% (95% confidence interval 81-88%) on a per-patient-based analysis. The area under the receiver-operating characteristic curve was 0.88. CONCLUSION: F-FDG PET/CT demonstrated high sensitivity for the diagnosis of patients with FUO. F-FDG PET/CT is an accurate technique in this setting, but the possibility of false-positive results should be kept in mind. This meta-analysis demonstrated the value of F-FDG PET/CT in the diagnosis of patients with FUO.

Diagnostic role of 18F-dihydroxyphenylalanine positron emission tomography in patients with congenital hyperinsulinism: a meta-analysis.

OBJECTIVES: Studies have reported the applications of F-dihydroxyphenylalanine (F-DOPA) PET in patients with congenital hyperinsulinism (CHI). The aim of this study was to systematically review and perform a meta-analysis of published data on the diagnostic role of F-DOPA PET in patients with CHI. MATERIALS AND METHODS: A comprehensive computer literature search of studies on F-DOPA PET or PET/computed tomography (CT) in patients with CHI was conducted. The pooled sensitivity and specificity of F-DOPA PET or PET/CT in patients with CHI were calculated. The area under the receiver-operating characteristic curve was calculated to measure the accuracy of F-DOPA PET or PET/CT in patients with CHI. RESULTS: Ten studies comprising 181 patients with CHI were included in this meta-analysis. The pooled sensitivity of F-DOPA PET and PET/CT in detecting CHI was 88% on a per-patient-based analysis. The pooled specificity of F-DOPA PET and PET/CT in demonstrating CHI was 79%. The area under the receiver-operating characteristic curve was 0.92 on a per-patient-based analysis. CONCLUSION: In patients with CHI, F-DOPA PET or PET/CT demonstrated high sensitivity and specificity. F-DOPA PET and PET/CT are accurate methods for the diagnosis of CHI. Nevertheless, possible sources of false-positive and false-negative results should be kept in mind.

18F-DOPA positron emission tomography/computed tomography application in congenital hyperinsulinism.

Congenital hyperinsulinism (CHI) during infancy is characterized by inappropriate insulin secretion resulting in persistent hypoglycemia. This can lead to irreversible severe neurological damage in the infant. There are two main histologic subtypes: diffuse and focal, both of which may require different surgical strategies. It is very important to differentiate focal leisons from diffuse leisons. However, the differentiation of diffuse leisons from focal leisons is challenging. Affected pancreatic areas utilize dihydroxyphenylalanine (DOPA) at a higher rate than normal pancreatic tissues; thus, labeling of L-DOPA with fluorine-18 (18F-DOPA) allows functional mapping of hyperinsulinism using positron emission tomography/computed tomography (PET/CT). In this article, we reviewed the 18F-DOPA PET/CT application in CHI. The aim of this review is to enhance the recognition of 18F-DOPA PET/CT application in the diagnosis of CHI.

Brown adipose tissue: distribution and influencing factors on FDG PET/CT scan.

Brown adipose tissue (BAT) functions as a thermogenic organ by producing heat to maintain body temperature in many mammals, especially in the young. BAT is generally found in deep cervical, supraclavicular, interscapular, and paravertebral regions, as well as areas near large vessels. If not recognized, BAT activity can considerably interfere with 18F-fluorodeoxyglucose position emission tomography (PET)/computed tomography (CT) image interpretation. BAT activation can be influenced by several factors and reduced by different methods. In this paper, we review BAT distribution and factors influencing BAT activity on FDG PET/CT scan. The purpose of this review is to enhance the recognition of pediatric-related physician of BAT on FDG PET/CT scan.

Primary osteogenic sarcoma of breast detected on Tc-99m MIBI scintigraphy and Tc-99m MDP skeletal scintigraphy.

A 64-year-old woman presented with a painless breast mass. Tc-99m methoxyisobutylisonitrile scintigraphy of both breasts showed a local area of abnormal uptake in the left breast in 5 min and 2 h. A skeletal scan showed very intense concentration of activity in the primary breast tumor in the left breast. A left mastectomy and an axillary dissection were performed. The predominant histologic type of the mass was an osteosarcoma, and the diagnosis of a primary osteogenic sarcoma of the breast was made. Primary osteogenic sarcoma of the breast is rare and represents less than 1% of all primary breast malignancies.