A Novel Multi-Epitope Vaccine Based on Urate Transporter 1 Alleviates Streptozotocin-Induced Diabetes by Producing Anti-URAT1 Antibody and an Immunomodulatory Effect in C57BL/6J Mice.

Hyperuricemia (HUA) is related to diabetes. Uric acid-induced inflammation and oxidative stress are risk factors for diabetes and its complications. Human urate transporter 1 (URAT1) regulates the renal tubular reabsorption of uric acid. IA-2(5)-P2-1, a potent immunogenic carrier designed by our laboratory, can induce high-titer specific antibodies when it carries a B cell epitope, such as B cell epitopes of DPP4 (Dipeptidyl peptidase-4), xanthine oxidase. In this report, we describe a novel multi-epitope vaccine composing a peptide of URAT1, an anti-diabetic B epitope of insulinoma antigen-2(IA-2) and a Th2 epitope (P2:IPALDSLTPANED) of P277 peptide in human heat shock protein 60 (HSP60). Immunization with the multi-epitope vaccine in streptozotocin-induced diabetes C57BL/6J mice successfully induced specific anti-URAT1 antibody, which inhibited URAT1 action and uric acid reabsorption, and increased pancreatic insulin level with a lower insulitis incidence. Vaccination with U-IA-2(5)-P2-1 (UIP-1) significantly reduced blood glucose and uric acid level, increased Th2 cytokines interleukin (IL)-10 and IL-4, and regulated immune reactions through a balanced Th1/Th2 ratio. These results demonstrate that the URAT1-based multi-epitope peptide vaccine may be a suitable therapeutic approach for diabetes and its complications.

The utility of diffusion-weighted imaging for differentiation of phyllodes tumor from fibroadenoma and breast cancer.

Objective: To evaluate the utility of apparent diffusion coefficient (ADC) values for differentiating breast tumors. Methods: The medical records of 17 patients with phyllodes tumor [PT; circular regions of interest (ROI-cs) n = 171], 74 patients with fibroadenomas (FAs; ROI-cs, n = 94), and 57 patients with breast cancers (BCs; ROI-cs, n = 104) confirmed by surgical pathology were retrospectively reviewed. Results: There were significant differences between PTs, FAs, and BCs in ADCmean, ADCmax, and ADCmin values. The cutoff ADCmean for differentiating PTs from FAs was 1.435 × 10-3 mm2/s, PTs from BCs was 1.100 × 10-3 mm2/s, and FAs from BCs was 0.925 × 10-3 mm2/s. There were significant differences between benign PTs, borderline PTs, and malignant PTs in ADCmean, ADCmax, and ADCmin values. The cutoff ADCmean for differentiating benign PTs from borderline PTs was 1.215 × 10-3 mm2/s, and borderline PTs from malignant PTs was 1.665 × 10-3 mm2/s. Conclusion: DWI provides quantitative information that can help distinguish breast tumors.

Micro- and macro-changes in early-stage type 2 diabetes mellitus without cognitive impairment: a diffusion tensor imaging (DTI) and surface-based morphometry (SBM) study.

Introduction: Brain structure and function changes are considered major brain damages in type 2 diabetes mellitus (T2DM), which likely has a close relationship with cognitive impairment. Many previous studies have shown by using brain structural and functional magnetic resonance imaging (MRI) methods that brain white and gray matter are damaged in T2DM, leading to cognitive impairment. Researches neglected patients of T2DM without cognitive dysfunction might also have brain changes. Methods: In this study, subjects with early stage T2DM with no cognitive dysfunction were enrolled to detect brain damages using the tract-based spatial statistics analysis (TBSS) method to demonstrate white matter (WM) micro changes and surface-based morphometry (SBM) method to assess cerebral cortex macro changes. Results: The whole-brain TBSS analysis revealed that there were no statistically significant changes in fractional anisotropy (FA) and mean diffusivity (MD), but the FA declined in some area of cerebral WM (p < 0.1). The SBM results showed no changes in cortical thickness (CT), cortical volume (CV), surface area (SA), and cortical sulcal curve (CSC) between these two groups, but pial local gyration index (LGI) was decreased in the precuneus (-log10, p = -3.327). Discussion: In conclusion, early stage T2DM patients without cognitive impairment had brain micro and macro structural damages, suggesting the potential use of MRI as an imaging marker to detect brain changes in early stage T2DM, which could not be observed and assessed clinically.

Trisodium scandium bis-(orthoborate).

Single crystals of tris-odium scandium bis-(orthoborate), Na(3)Sc(BO(3))(2), have been obtained by spontaneous crystallization from an Na(2)O-Sc(2)O(3)-B(2)O(3) melt. The crystal structure features a three-dimensional framework composed of planar [BO(3)](3-) groups and distorted ScO(6) octa-hedra with Na atoms in the cavities. The Sc atom occupies a special position (Wyckoff position 2b, site symmetry -1) and of the two Na atoms, one occupies a special position (Wyckoff position 2c, site symmetry -1).

Diagnosis and Treatment to a Post-Craniotomy Intracranial Infection Caused by Corynebacterium.

Objective: To explore the perioperative prophylactic medication, identification of Causative pathogen and the treatment strategy of post-craniotomy intracranial infection (PCII) caused by Corynebacterium. Patients and Methods: A 47-year-old overweight male patient with hypertension, diabetes, cerebral hemorrhage and recalcitrant syphilis was clinically diagnosed with PCII based on cerebrospinal fluid (CSF) routine examination (RT), biochemical test (BT), neuroimaging CT and MRI scans, bacterial culture and identification of CSF and clinical manifestations. The risk factors of PCII and perioperative antibiotic prophylaxis were analyzed based on some reviews. The identification of the Corynebacterium Jeikeium (C. Jeikeium) and Corynebacterium simulans (C. simulans) was confirmed by CSF bacterial culture, antibiotics sensitivity in vitro and Metagenomic next-generation sequencing (mNGS) of pathogenic microorganisms, respectively. In addition, individualized therapy schemes were modified according to antimicrobial susceptibility of pathogens and mNGS of pathogenic microorganisms combined with the pathologic and physiological conditions of patients. The efficacy was evaluated depending on the changes in patients' body temperature, clinical manifestation, CSF RT, BT, and other infection-related indicators. Results: The patient recovered after 5 weeks of individualized comprehensive treatment and was discharged home, no recurrence had been observed for three months. Conclusion: This is likely the first reported case of chronic PCII caused by two species of Corynebacterium simultaneously in high risk patient. The PCII can not be prevented by the perioperative antibiotic prophylaxis recommended by the guidelines, prophylaxis need to be individualized based on the risk of infection and the colonization status of the patient. Causative pathogens can be identified by CSF culture and mNGS of pathogenic microorganisms. A judicious antimicrobial therapy plan should take into account not only the in vitro antimicrobial susceptibility, but also the penetration of the antimicrobial agent into the cerebrospinal fluid. It was an excellent choice to combine intrathecal vancomycin with intravenous linezolid to treat PCII resulted from Corynebacterium.

Apparent diffusion coefficient histogram analysis for differentiating solid ovarian tumors.

Objective: To evaluate the utility of apparent diffusion coefficient (ADC) histogram analysis to differentiate between three types of solid ovarian tumors: granulosa cell tumors (GCTs) of the ovary, ovarian fibromas, and high-grade serous ovarian carcinomas (HGSOCs). Methods: The medical records of 11 patients with GCTs of the ovary (regions of interest [ROI-cs], 137), 61 patients with ovarian fibromas (ROI-cs, 161), and 14 patients with HGSOCs (ROI-cs, 113) confirmed at surgery and histology who underwent diffusion-weighted imaging were retrospectively reviewed. Histogram parameters of ADC maps (ADCmean, ADCmax, ADCmin) were estimated and compared using the Kruskal-WallisH test and Mann-Whitney U test. The area under the curve of receiver operating characteristic curves was used to assess the diagnostic performance of ADC parameters for solid ovarian tumors. Results: There were significant differences in ADCmean, ADCmax and ADCmin values between GCTs of the ovary, ovarian fibromas, and HGSOCs. The cutoff ADCmean value for differentiating a GCT of the ovary from an ovarian fibroma was 0.95×10-3 mm2/s, for differentiating a GCT of the ovary from an HGSOC was 0.69×10-3 mm2/s, and for differentiating an ovarian fibroma from an HGSOC was 1.24×10-3 mm2/s. Conclusion: ADCmean derived from ADC histogram analysis provided quantitative information that allowed accurate differentiation of GCTs of the ovary, ovarian fibromas, and HGSOCs before surgery.

Development of a chiral HPLC method for the separation and quantification of hydroxychloroquine enantiomers.

Hydroxychloroquine (2-[[4-[(7-Chloroquinolin-4-yl) amino]pentyl](ethyl) amino]-ethanol, HCQ), an effective anti-malarial drug, has been tested in the clinics for potential treatment of severe coronavirus disease 2019 (COVID-19). Despite the controversy around the clinical benefits of HCQ, the existence of a chiral center in the molecule to possess two optical isomers suggests that there might be an enantiomeric difference on the treatment of COVID-19. Due to their poor resolution and the inability of quantification by previously reported methods for the analysis of HCQ enantiomers, it is necessary to develop an analytical method to achieve baseline separation for quantitative and accurate determination of the enantiomeric purity in order to compare the efficacy and toxicity profiles of different enantiomers. In this study, we developed and validated an accurate and reproducible normal phase chiral high-performance liquid chromatography (HPLC) method for the analysis of two enantiomers of HCQ, and the method was further evaluated with biological samples. With this newly developed method, the relative standard deviations of all analytes were lower than 5%, and the limits of quantification were 0.27 µg/ml, 0.34 µg/ml and 0.20 µg/ml for racemate, R- and S-enantiomer, respectively. The present method provides an essential analytical tool for preclinical and clinical evaluation of HCQ enantiomers for potential treatment of COVID-19.

Imaging in testicular epidermoid cysts.

OBJECTIVE: To evaluate the utility of findings on ultrasound and magnetic resonance imaging (MRI) for the preoperative diagnosis of testicular epidermoid cysts (TEC). METHODS: The medical records of five patients treated for TEC at our institution between July 2010 and May 2017 were retrospectively reviewed. RESULTS: Imaging revealed a target or onion skin appearance on ultrasonography and MRI. Pathological examinations showed "bread slag-like" materials within the TEC. Lesions failed to demonstrate enhancement after Gd-DTPA injection. CONCLUSION: In the cases of TEC in the current study, accurate diagnosis ensured enucleation of the testicular cyst was performed rather than testicular resection.

A novel IgG1 monoclonal antibody against xanthine oxidase alleviates inflammation induced by potassium oxonate in mice.

Xanthine oxidase (XOD) is a key enzyme that catalyzes xanthine to uric acid. Most of the urate-lowering medicines targeting XOD have a limited effect on alleviating inflammation in spite of significant effects on decreasing serum uric acid level. In this study, we produced and characterized a novel monoclonal antibody (Anti-XOD mAb) using hybridoma technology based on a novel peptide OI5P-1(O-IA2(5)-P2-1),which containing a B-cell epitope of XOD and a novel Th2 built-in adjuvant I5P-1(IA2(5)-P2-1). Results of western blotting and cross-reactivity assay indicated that the mAb binds specifically to XOD and the affinity was 2.523×1010L/mol. The mAb reduced serum uric acid level and hepatic xanthine oxidase activity in potassium oxonate induced mice. A decreased methane dicarboxylic aldehyde level and an improved superoxide dismutase level in mAb treated mice indicated anti-lipid peroxidation effects of the mAb. Moreover, the mAb showed a significant immunomodulatory effect which could shift Th1/Th2 balance to Th2-dominant immunity. The mAb treatment alleviates inflammation induced by potassium oxonate, superior to the small molecule allopurinol treatment. For the first time, these results showed that the anti-XOD mAb may serve as a promising therapeutic approach for inflammatory response related to uric acid.

A novel multi-epitope vaccine based on Dipeptidyl Peptidase 4 prevents streptozotocin-induced diabetes by producing anti-DPP4 antibody and immunomodulatory effect in C57BL/6J mice.

Type 1 diabetes is a chronic organ-specific autoimmune disease in which selective destruction of insulin-producing ß-cells leads to impaired glucose metabolism and its attendant complications. A series of Dipeptidyl peptidase 4 (DPP4) inhibitors have been developed and granted approval in the treatment of type 2 diabetes mellitus by inhibiting the enzymatic degradation of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP). An increasing number of studies have shown the potential benefits of DPP4 inhibitors for type 1 diabetes. In this report, we describe a novel multi-epitope vaccine comprising a B cell epitope of DPP4, an anti-diabetic B cell epitope of Insulinoma antigen-2 (IA-2) and a Th2 epitope of P277 peptide in human heat shock protein 60 (HSP60). Immunization with the multi-epitope vaccine in streptozotocin (STZ) treated mice successfully induced specific anti-DPP4 antibody and increased serum GLP-1 level. Moreover, this antibody lasted for more than 7 weeks. Inoculation of this vaccine in C57BL/6J mice significantly reduced blood glucose level, improved glucose excursion and increased plasma insulin concentration. Consistent with a lower diabetic and insulitis incidence, induced splenic T cell proliferation and tolerance were observed. IFN-γ and IL-2 secretion reduced, but IL-10 and IL-4 increased significantly in the Dipeptidyl Peptidase 41-Insulinoma antigen-2(5)-P2-1 (D41-IP) treated mice compared to the Insulinoma antigen-2(5)-P2-1 (IA2(5)P2-1) and control group due to the potential immunomodulatory effect of the epitopes in the vaccine. Our results demonstrate that this multi-epitope vaccine may serve as a promising therapeutic approach against type 1 diabetes.

Hyaluronan-modified superparamagnetic iron oxide nanoparticles for bimodal breast cancer imaging and photothermal therapy.

Theranostic nanoparticles with both imaging and therapeutic abilities are highly promising in successful diagnosis and treatment of the most devastating cancers. In this study, the dual-modal imaging and photothermal effect of hyaluronan (HA)-modified superparamagnetic iron oxide nanoparticles (HA-SPIONs), which was developed in a previous study, were investigated for CD44 HA receptor-overexpressing breast cancer in both in vitro and in vivo experiments. Heat is found to be rapidly generated by near-infrared laser range irradiation of HA-SPIONs. When incubated with CD44 HA receptor-overexpressing MDA-MB-231 cells in vitro, HA-SPIONs exhibited significant specific cellular uptake and specific accumulation confirmed by Prussian blue staining. The in vitro and in vivo results of magnetic resonance imaging and photothermal ablation demonstrated that HA-SPIONs exhibited significant negative contrast enhancement on T2-weighted magnetic resonance imaging and photothermal effect targeted CD44 HA receptor-overexpressing breast cancer. All these results indicated that HA-SPIONs have great potential for effective diagnosis and treatment of cancer.

Glycosaminoglycan-targeted iron oxide nanoparticles for magnetic resonance imaging of liver carcinoma.

To develop an efficient probe for targeted magnetic resonance (MR) imaging of liver carcinoma, the surface modification of superparamagnetic iron oxide nanoparticles (SPIONs) was carried out by conjugating a naturally-occurring glycosaminoglycan with specific biological recognition to human hepatocellular liver carcinoma (HepG2) cells. These modified SPIOs have good water dispersibility, superparamagnetic property, cytocompatibility and high magnetic relaxivity for MR imaging. When incubated with HepG2 cells, they demonstrated significant cellular uptake and specific accumulation, as confirmed by Prussian blue staining and confocal microscopy. The in vitro MR imaging of HepG2 cells and in vivo MR imaging of HepG2 tumors confirmed their effectiveness for targeted MR imaging of liver carcinoma.