Molecular imaging reveals the heterogeneous progression of tumor cells and tumor stroma: a practice of FDG PET and FAPI PET in diagnosing PSMA-negative bone metastases of progressive prostate cancer.

Tumors are often with complex and heterogeneous biological processes, such as glycometabolism and fibrosis, which are the main biochemical pathways that determine therapeutic effects. Specifically, this study aims to assess the diagnosing performance of 18F-FDG and 68Ga-FAPI-04 PET for different stages of progressive bone metastases with PSMA-negative pathology. Bone metastatic mouse model of prostate cancer was constructed via intra-bone injection of PSMA-negative prostate cancer PC3 cells. Cellular uptakes of 18F-FDG and 68Ga-FAPI-04 were separately performed on PC3, NIH-3T3 (FAP-positive) and a mixture. 68Ga-PSMA-11, 18F-FDG and 68Ga-FAPI-04 PET/CT imaging were performed at 2, 4 weeks after tumor cell transplantation. Furthermore, PSMA and FAP expression in bone metastases were assessed by immunohistochemistry, and then compared with the imageological findings. On the cellular level, the independent tracer uptake on the basis of glycometabolism and fibrosis was observed. For animal imaging, 68Ga-PSMA-11 imaging showed weak or absent tracer uptake in PSMA-negative bone metastatic lesions. In contrast, 68Ga-FAPI-04 PET of bone metastases had a higher uptake and tumor-to-muscle (T/M) ratio than 18F-FDG PET that was relative steady during the observation, but T/M ratio of fibrosis gradually decreased with increasing tumor growth, which ranged from 5.11 ± 1.26 at 2 weeks to 3.54 ± 0.23 at 4 weeks, revealing the delayed formation of tumor stroma in rapid proliferation. In addition, PET imaging results were corroborated by immunohistochemical assessment. In conclusion, molecular imaging approach revealed the heterogeneous progression of tumor cells and tumor stroma of bone metastasis of prostate cancer, and further confirming the necessity of multi-molecular imaging in cancer imaging.

A strategy to improve the solubility and bioavailability of the insoluble drug piperlongumine through albumin nanoparticles.

Piperlongumine (PL) is a biologically active alkaloid derived from peppers, has significant cytotoxic effects on cancer with no cytotoxicity. This study used NabTM technology to prepare PL albumin nanoparticles (PL-BSA-NPs) to improve water solubility and bioavailability. We carried out a pharmacological evaluation of the PL-BSA-NPs. The morphological profile of the PL-BSA-NPs was relatively uniform, with an average particle size of approximately 210 nm, with drug load of 2.1% and encapsulation rate of 87.6%. PL-BSA-NPs were stable for 4 weeks when stored at 4°C. In vitro release behavior of the PL-BSA-NPs showed a sustained release, with a cumulative release of 67.24% in approximately 24 hours. The pharmacokinetic properties of PL-BSA-NPs were shown that PL-BSA-NPs could maintain a certain level of blood drug concentration for a long time, thus demonstrating the sustained release and increased bioavailability of PL. Finally, we investigated the in vitro antitumor activity of the PL-BSA-NPs and found that PL can significantly inhibit HepG2 cell proliferation, and that PL-BSA-NPs enhanced the inhibitory effect of PL on this proliferative effect. Thus, we concluded that PL can destroy liver cancer cells by increasing ROS levels. These results suggested that PL-BSA-NPs show promising potential as a targeted anti-tumor drug.

PD-L1 ImmunoPET on the basis of Avidin/Biotin pre-targeted cancer imaging.

This study aimed to establish the radio-immune imaging protocol on the basis of Avidin/Biotin system. The programmed death-ligand 1 (PD-L1) antibody (Atezolizumab) was employed as the primary molecule in targeting PD-L1, and the two-step strategy, consisting of the first injection of Avidin-conjugated PD-L1 monoclonal antibody (Atezolizumab) and the second injection of 7.4 MBq 68Ga-Biotin with a 60 h interval, was then verified on the colon cancer-bearing mice. PET imaging was performed at 30, 90, 180 min to measure the standard uptake value and tumor to liver ratios. Cellular binding experiments and in vivo distribution showed that the conjugation of Avidin did not affect the affinity of Atezolizumab to PD-L1 antigen. Biotin was radio-labeled with 68Ga with radiolabeling efficiency of 70.5 ± 3.5% and purification was needed to increase the radiochemical purity. For PD-L1-positive tumors, SUVmax was 0.38 ± 0.06 in the Avidin-Atezolizumab pre-treated mice at 90 min; the tumor/liver ratios of pre-targeting group were 1.06 ± 0.19 and 0.97 ± 0.16 at 30 and 90 min, while the absence of pre-treatment of Avidin was of the lower ratios as 0.88 ± 0.01 and 0.54 ± 0.11 when 68Ga-Biotin served as the radiopharmaceutical as well. In conclusion, pre-targeting immunoPET strategy can elevate the target-to-nontarget ratio, decrease the blood background and shorten the interval between injection of radiopharmaceuticals and PET scan, providing a highly PD-L1-specific and sensitive imaging method for the detection of tumorous immune micro-environment.

CircPalm2 knockdown alleviates LPS-evoked pulmonary microvascular endothelial cell apoptosis and inflammation via miR-450b-5p/ROCK1 axis.

BACKGROUND: Emerging evidence has revealed that circular RNAs (circRNAs) have roles in regulating the complex pathologies of sepsis-induced acute lung injury (sepsis-ALI). Herein, this work aimed to investigate the potential role and mechanism of circPalm2 in the process of sepsis-ALI. METHODS: Primary mice pulmonary microvascular endothelial cells (MPVECs) in functional group were exposed to lipopolysaccharide (LPS). Levels of genes and proteins were measured by qRT-PCR and western blotting. Functional experiments were conducted using In vitro functional experiments were conducted using cell counting kit-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry and ELISA analysis. The binding between miR-450b-5p and circPalm2 or ROCK1 (Rho Associated Coiled-Coil Containing Protein Kinase 1) was validated using dual-luciferase reporter, RNA immunoprecipitation and pull-down assays. RESULTS: LPS treatment caused the increase of circPalm2 and ROCK1, as well as the decrease of miR-450b-5p in MPVECs. Knockdown of circPalm2 attenuated LPS-induced proliferation arrest, apoptosis, and production of proinflammatory cytokine IL-6, IL-ß and TNF-α in MPVECs. Mechanistically, circPalm2 sequestered miR-450b-5p to up-regulate ROCK1 expression, revealing the circPalm2/miR-450b-5p/ROCK1 feedback loop. Moreover, the protective functions mediated by circPalm2 silencing on MPVECs under LPS exposure were abolished by miR-129-5p inhibition or ROCK1 overexpression. CONCLUSION: CircPalm2 knockdown can alleviate LPS-evoked MPVEC apoptosis and inflammation via miR-450b-5p/ROCK1 axis, suggesting the potential involvement of this ceRNA network in sepsis-ALI and a broader approach for the therapy of sepsis-ALI.

Study on the association between the polymorphism of MCP-1 rs1024611 and the genetic susceptibility of type 2 diabetes with sepsis.

Monocyte chemoattractant protein-1 (MCP-1) rs1024611 (-2518 A > G) polymorphism are associated with inflammatory diseases. In this study, we investigate the relationship between MCP-1 rs1024611 polymorphism and genetic susceptibility of type 2 diabetes mellitus (T2DM) with sepsis. Two hundred eighty-five patients with T2DM are divided into the diabetes with sepsis group (combined group, 113 cases) and the diabetes group (172 cases). Blood samples and corresponding clinical data were collected. MCP-1 rs1024611 polymorphism in blood samples was detected by pyrosequencing. Meanwhile, the expressions of MCP-1, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1ß, and IL-6 in blood samples were detected by real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The relationship between different genotypes of MCP-1 rs1024611 polymorphic locus and T2DM with sepsis was analyzed by combining with the clinical data of the patients. The frequencies of rs1024611 AG/GG genotypes and G allele in T2DM with sepsis group were significantly higher than those in T2DM patients without sepsis (P = .004 for AG/GG vs AA genotypes; P = .037 for G allele vs A allele). Subgroup analysis showed that the rs1024611 G allele frequency in the septic shock group was significantly higher than the general sepsis group (P = .02). The expressions of MCP-1 and TNF-α in GG genotypes in T2DM with sepsis group were significantly higher than AA or GA genotypes (P < .05). This study preliminarily showed that the rs1024611 A > G polymorphism within the promoter region of MCP-1 gene can upregulate the expression of MCP-1 gene and proinflammatory cytokine TNF-α, which ultimately contributed to the predisposition and progression of T2DM with sepsis.

Detection value of FOXO1 gene methylation, blood glucose and lipids in patients with type 2 diabetic kidney disease.

Forkhead transcription factor O1 (FOXO1) methylation is associated with inflammation. Diabetic kidney disease (DKD) is characterized with increased inflammatory markers such as uric acid, hemogram indices, C-reactive protein derived markers, omentin and neuregulin. This study aimed to investigate the effect of DNA methylation in FOXO1 gene promoter, blood glucose and lipids in the process of type 2 DKD. Bisulfite genomic sequencing was used to monitor DNA methylation in the promoter region (+1021, +1193) of FOXO1 gene. The detections were taken in glycosylated hemoglobin A1c, fasting plasma glucose and blood lipid. 81 participants were divided into the control group, the preliminary diabetes mellitus group, the pure diabetes mellitus group, and the DKD group. The other groups displayed higher fasting plasma glucose than the control group (all P value < .05). The fasting plasma glucose level was higher in the pure diabetes mellitus group than the preliminary diabetes mellitus group (P = .004). The levels of HbA1c were higher in other groups than control group and preliminary diabetes mellitus groups (all P values < .01). The high-density lipoprotein level was lower in the DKD group (P = .021, P = .022) than control and pure diabetes mellitus group. The levels of low-density lipoprotein were statistically lower in preliminary diabetes mellitus and DKD groups than control group (all P value < .02). Along with the progress of DKD, a down trend was observed in the total methylation rate of FOXO1 gene (P = .025), which contains 5 CpG sites (1021, +1193) in the promoter. Hypomethylation in the promoter of FOXO1 gene, hyperglycemia and low level of serum lipid might be associated with the pathogenesis of type 2 DKD.

Multiple endocrine neoplasia 2A with RET mutation p.Cys611Tyr: A case report.

RATIONALE: Multiple endocrine neoplasia 2A (MEN2A) is a rare autosomal-dominant genetic syndrome, frequently misdiagnosed or neglected clinically, resulting in delayed therapy to patients. PATIENT CONCERNS: A 47-year-old Chinese male patient underwent laparoscopic right adrenal tumorectomy, and postoperative pathology confirmed the tumor as pheochromocytoma (PHEO). He was readmitted to the department of endocrinology and metabolism due to constant increase in carcinoembryonic antigen (CEA) at 5 months after the operation. DIAGNOSIS: The patient was confirmed with medullary thyroid carcinoma (MTC), multiple neck lymph node metastasis, and pituitary microadenoma. The p.Cys611Tyr (c.1832G>A, C611Y) mutation was detected. Therefore, he was diagnosed with MEN2A. INTERVENTIONS: He underwent total thyroidectomy. The gene-sequencing analysis of his family was conducted, and the C611Y mutation was detected in his daughter. OUTCOMES: The level of carcinoembryonic antigen decreased significantly after thyroidectomy in this patient. Long-term follow-up management was conducted. Elevated serum calcitonin and bilateral thyroid nodules were found in his 13-year-old daughter. Thus, MEN2A was highly suspected and she was suggested to undergo total thyroidectomy. CONCLUSION: Patients with MEN2A should be screened regularly and managed by a multidisciplinary team.

A pH-sensitive and sustained-release oral drug delivery system: the synthesis, characterization, adsorption and release of the xanthan gum-graft-poly(acrylic acid)/GO-DCFP composite hydrogel.

In this study, graphene oxide (GO) was successfully prepared using the improved Hummers method, and the prepared GO powder was dissolved in distilled water and subjected to ultrasonic stripping. Diclofenac potassium (DCFP) was selected as a model drug to systematically evaluate the adsorption mechanism of DCFP by GO. Different reaction models were constructed to fit the adsorption kinetics and adsorption isotherms of DCFP on GO, in order to further explore the underlying adsorption mechanism. The results demonstrated that the pseudo-second-order kinetic model and Freundlich model could better delineate the adsorption process of DCFP by GO. Both π-π stacking and hydrophobic interaction were mainly involved in the adsorption process, and there were electrostatic interaction and hydrogen bonding at the same time. Then, the xanthan gum-graft-poly(acrylic acid)/GO (XG-g-PAA/GO) composite hydrogel was synthesized by in situ polymerization as a slow-release drug carrier. For this reason, a XG-g-PAA/GO-DCFP composite hydrogel was synthesized, and its in vitro drug release and pharmacokinetic data were assessed. The results showed that the synthesized XG-g-PAA/GO composite hydrogel had a certain mechanical strength and uniform color, indicating that GO is evenly distributed in this composite hydrogel. Moreover, the results of a swelling ratio test demonstrated that the swelling ratios of the XG-g-PAA/GO composite hydrogel were significantly increased with increasing pH values, implying that this material is sensitive to pH. The in vitro drug release experiment showed that the cumulative release of DCFP after 96 h was significantly higher in artificial intestinal fluid than in artificial gastric fluid. These findings indicate that the XG-g-PAA/GO-DCFP composite hydrogel exhibits pH sensitivity under physiological conditions. Besides, the results of in vivo pharmacokinetic analysis revealed that the t 1/2 of DCFP group was 2.03 ± 0.35 h, while that of the XG-g-PAA/GO-DCFP composite hydrogel group was 10.71 ± 2.04 h, indicating that the synthesized hydrogel could effectively prolong the drug action time. Furthermore, the AUC(0-t) of the DCFP group was 53.99 ± 3.18 mg L-1 h-1, while that of the XG-g-PAA/GO-DCFP composite hydrogel group was 116.79 ± 14.72 mg L-1 h-1, suggesting that the bioavailability of DCFP is greatly enhanced by this composite hydrogel. In conclusion, this study highlights that the XG-g-PAA/GO-DCFP composite hydrogel can be applied as a sustained-release drug carrier.

Perturbation of secretory Ig A in saliva and its daily variation by academic stress.

OBJECTIVES: Several studies have reported that the secretory immunoglobulin A (S-IgA) concentration in saliva is an indicator of psychological stress. The aim of this study was to clarify the relationship between S-IgA and the stress from academic examinations. METHODS: S-IgA levels in 10 medical student volunteers from the second year course between May 4 and July 13, 2000 were examined using the ELISA method. RESULTS: There was a tendency for S-IgA in saliva to be higher on the day before academic examinations and during them, and lower on the days between these examinations. CONCLUSIONS: It may be possible to use this measurement to monitor psychological stress in students and workers.