Showing papers by "Xuzhou Medical College published in 2023"

Lateral septum-lateral hypothalamus circuit dysfunction in comorbid pain and anxiety

Abstract: Pain and anxiety comorbidities are a common health problem, but the neural mechanisms underlying comorbidity remain unclear. We propose that comorbidity implies that similar brain regions and neural circuits, with the lateral septum (LS) as a major candidate, process pain and anxiety. From results of behavioral and neurophysiological experiments combined with selective LS manipulation in mice, we find that LS GABAergic neurons were critical for both pain and anxiety. Selective activation of LS GABAergic neurons induced hyperalgesia and anxiety-like behaviors. In contrast, selective inhibition of LS GABAergic neurons reduced nocifensive withdrawal responses and anxiety-like behaviors. This was found in two mouse models, one for chronic inflammatory pain (induced by complete Freund's adjuvant) and one for anxiety (induced by chronic restraint stress). Additionally, using TetTag chemogenetics to functionally mark LS neurons, we found that activation of LS neurons by acute pain stimulation could induce anxiety-like behaviors and vice versa. Furthermore, we show that LS GABAergic projection to the lateral hypothalamus (LH) plays an important role in the regulation of pain and anxiety comorbidities. Our study revealed that LS GABAergic neurons, and especially the LSGABAergic-LH circuit, are a critical to the modulation of pain and anxiety comorbidities.

Mid- to long-term outcomes of initial transjugular intrahepatic portosystemic shunt versus anticoagulation for pyrrolizidine alkaloid-induced hepatic sinusoidal obstruction syndrome

Abstract: Anticoagulation therapy (AT) is often used as the initial treatment for pyrrolizidine alkaloid (PA)-induced hepatic sinusoidal obstruction syndrome (HSOS). However, transjugular intrahepatic portosystemic shunt (TIPS) is an alternative treatment. This study aimed to determine the mid- to long-term outcomes of TIPS versus AT as the initial treatment for PA-induced HSOS.We retrospectively analyzed the clinical data of 61 patients with PA-induced HSOS that were collected between November 2015 and July 2021. The patients were allocated to the TIPS group ( n = 20) or the AT group ( n = 41). These two groups were divided into subgroups according to the severity grading. The clinical data of the patients in both groups were analyzed. Cumulative survival rates were calculated and compared between the two groups and among the subgroups.The clinical symptoms and signs improved or stabilized in 100% of the patients following TIPS and in 85% of the patients following AT at discharge ( P = 0.166). The mortality rate was 0.0% in the TIPS group and 34.1% in the AT group ( P = 0.005). The patients were followed up for 2-69 months (mean, 26.3 ± 20.5 months). In the mild- and moderate-grade subgroups, there was no difference in the cumulative survival rate between the TIPS and AT groups ( P = 0.589 and P = 0.364, respectively). In the severe and very severe-grade subgroups, the cumulative survival rate was higher in the TIPS group than in the AT group ( P = 0.018 and P = 0.025, respectively).AT is a suitable initial treatment for mild or moderate PA-induced HSOS, whereas TIPS should be considered the appropriate initial treatment for severe or very severe PA-induced HSOS.

Robo4 inhibits gamma radiation-induced permeability of a murine microvascular endothelial cell by regulating the junctions

Abstract: Hematopoietic stem cell transplantation involves irradiation preconditioning which causes bone marrow endothelial cell dysfunction. While much emphasis is on the reconstitution of hematopoietic stem cells in the bone marrow microenvironment, endothelial cell preservation is indispensable to overcome the preconditioning damages. This study aims to ascertain the role of Roundabout 4 (Robo4) in regulating irradiation-induced damage to the endothelium.Microvascular endothelial cells were treated with γ-radiation to establish an endothelial cell injury model. Robo4 expression in the endothelial cells was manipulated employing lentiviral-mediated RNAi and gene overexpression technology before irradiation treatment. The permeability of endothelial cells was measured using qPCR, immunocytochemistry, and immunoblotting to analyze the effect on the expression and distribution of junctional molecules, adherens junctions, tight junctions, and gap junctions. Using Transwell endothelial monolayer staining, FITC-Dextran permeability, and gap junction-mediated intercellular communication (GJIC) assays, we determined the changes in endothelial functions after Robo4 gene manipulation and irradiation. Moreover, we measured the proportion of CD31 expression in endothelial cells by flow cytometry. We analyzed variations between two or multiple groups using Student's t-tests and ANOVA.Ionizing radiation upregulates Robo4 expression but disrupts endothelial junctional molecules. Robo4 deletion causes further degradation of endothelial junctions hence increasing the permeability of the endothelial cell monolayer. Robo4 knockdown in microvascular endothelial cells increases the degradation and delocalization of ZO-1, PECAM-1, occludin, and claudin-5 molecules after irradiation. Conversely, connexin 43 expression increases after silencing Robo4 in endothelial cells to induce permeability but are readily destroyed when exposed to 10 Gy of gamma radiation. Also, Robo4 knockdown enhances Y731-VE-cadherin phosphorylation leading to the depletion and destabilization of VE-cadherin at the endothelial junctions following irradiation. However, Robo4 overexpression mitigates irradiation-induced degradation of tight junctional proteins and stabilizes claudin-5 and ZO-1 distribution. Finally, the enhanced expression of Robo4 ameliorates the irradiation-induced depletion of VE-cadherin and connexin 43, improves the integrity of microvascular endothelial cell junctions, and decreases permeability.This study reveals that Robo4 maintains microvascular integrity after radiation preconditioning treatment by regulating endothelial permeability and protecting endothelial functions. Our results also provided a potential mechanism to repair the bone marrow vascular niche after irradiation by modulating Robo4 expression.

Hypoxia-induced ROS aggravate tumor progression through HIF-1α-SERPINE1 signaling in glioblastoma

Abstract: Hypoxia, as an important hallmark of the tumor microenvironment, is a major cause of oxidative stress and plays a central role in various malignant tumors, including glioblastoma. Elevated reactive oxygen species (ROS) in a hypoxic microenvironment promote glioblastoma progression; however, the underlying mechanism has not been clarified. Herein, we found that hypoxia promoted ROS production, and the proliferation, migration, and invasion of glioblastoma cells, while this promotion was restrained by ROS scavengers N-acetyl-L-cysteine (NAC) and diphenyleneiodonium chloride (DPI). Hypoxia-induced ROS activated hypoxia-inducible factor-1α (HIF-1α) signaling, which enhanced cell migration and invasion by epithelial-mesenchymal transition (EMT). Furthermore, the induction of serine protease inhibitor family E member 1 (SERPINE1) was ROS-dependent under hypoxia, and HIF-1α mediated SERPINE1 increase induced by ROS via binding to the SERPINE1 promoter region, thereby facilitating glioblastoma migration and invasion. Taken together, our data revealed that hypoxia-induced ROS reinforce the hypoxic adaptation of glioblastoma by driving the HIF-1α-SERPINE1 signaling pathway, and that targeting ROS may be a promising therapeutic strategy for glioblastoma.

Downregulation of microRNA-23a confers protection against myocardial ischemia/reperfusion injury by upregulating tissue factor pathway inhibitor 2 following luteolin pretreatment in rats

Abstract: To the Editor: Myocardial ischemia/reperfusion injury (MI/RI) often causes death, dysfunction, or damage to the cardiomyocytes, and thus more effective clinical treatments are required.[1] MicroRNAs (miRNAs) bind to target mRNAs to regulate post-transcriptional gene expression, with important effects on MI/RI by regulating critical molecular signaling pathways.[2] Previous studies have shown that flavonoids have beneficial effects on cardiac function and the outcomes after cardiac operations. Luteolin is a type of flavonoid, and high dietary intake of luteolin-rich food decreases the myocardial infarction area.[3] However, the transcriptional mechanism that allows luteolin to modulate gene expression and protect against MI/RI remains unknown. In this study, we screened the miRNAs with the greatest effects in MI/RI models after luteolin pretreatment. We identified the target gene of the miRNAs and explored the mechanisms associated with the protective effects of luteolin against MI/RI both in vitro and in vivo. All the experiments were performed in accordance with guidelines and approved by the Animal Ethics Committee of the Xuzhou Medical University (No. CMCACUC2014-04-027). Luteolin pretreatment can protect against MI/RI. The luteolin concentrations were 7.5 μmol/L for neonatal rat cardiomyocytes (NRCs) and 16.0 μmol/L for H9c2 cells according to detection by the methyl thiazoleterazolium assay [Supplementary Figure 1A, https://links.lww.com/CM9/B188]. Lactate dehydrogenase (LDH) detection and apoptosis rate assessments using the Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay showed that luteolin pretreatment decreased the LDH levels and inhibited cell apoptosis (all P < 0.01; Supplementary Figure 1B, C, https://links.lww.com/CM9/B188). Furthermore, cleaved caspase-3 was activated after simulated I/R injury, but activation was decreased by luteolin pretreatment (all P < 0.01; Supplementary Figure 1D, https://links.lww.com/CM9/B188). MiRNAs are potential therapeutic targets in various diseases.[4] Eight miRNAs (miRNA-1, -21, -23a, -24, -133a, -199a-3p, -214, and -378) were searched for in MEDLINE, PubMed Central, and the Human microRNA Disease Database. The reverse transcription polymerase chain reaction (RT-PCR) results showed that miRNA-1, -23a, -24, -133a, and -378 were upregulated in perfused rat hearts after MI/RI, but that luteolin pretreatment downregulated miRNA-23a and upregulated miR-378. MiRNA-1, -21, -23a, and -24 were upregulated in the simulate I/R (sI/R) model in NRCs, but luteolin pretreatment only downregulated miRNA-23a. Thus, miRNA-23a was selected for further investigation [Supplementary Figure 2, https://links.lww.com/CM9/B188]. Our in vitro experiments showed that miRNA-23a was upregulated after sI/R, but downregulated by luteolin pretreatment (all P < 0.01; Supplementary Figure 1E, https://links.lww.com/CM9/B188). The miRNA-23a mimic aggravated sI/R injury and activated the cleavage of caspase-3, whereas the miRNA-23a inhibitor had the opposite effect (all P < 0.01; Supplementary Figure 1F, G, https://links.lww.com/CM9/B188). Overexpression of miRNA-23a abolished the protective effect of luteolin and the inactivation effect on cleaved caspase-3 (all P > 0.05; Supplementary Figure 1H, I, https://links.lww.com/CM9/B188). Tissue factor pathway inhibitor 2 (TFPI2) acts as a tumor suppressor and modulates cell growth, apoptosis, and invasion.[5] TFPI2 is upregulated by ischemic preconditioning in rats, thereby suggesting that it may alleviate apoptosis. Using TargetScan, PicTar, and miRDB, we found that Tfpi2 is a putative target gene of miRNA-23a. Real-time quantitative PCR (qPCR) and Western blotting showed that the miRNA-23a mimic did not affect the mRNA level of Tfpi2 but decreased its protein expression level, whereas the miRNA-23a inhibitor had the opposite effect [Supplementary Figure 3A, B, https://links.lww.com/CM9/B188]. Luciferase reporter assays showed that overexpression of miRNA-23a decreased the luciferase activity (P < 0.001), whereas knockdown of miRNA-23a increased the luciferase activity (P < 0.01). Moreover, induced mutation of the predicted binding site of miRNA-23a abolished these effects (all P > 0.05; Supplementary Figure 3C, https://links.lww.com/CM9/B188). Western blotting analysis for the luteolin group showed that TFPI2 was upregulated in both NRCs and H9c2 cells (all P < 0.001, vs. sI/R group; Supplementary Figure 1J, https://links.lww.com/CM9/B188). Knockdown of TFPI2 with siRNA partly abolished the protective effects of luteolin [Supplementary Figure 1K,L, https://links.lww.com/CM9/B188]. After pretreatment with the miRNA-23a mimic, the TFPI2 protein and cleaved caspase-3 levels were almost the same with or without luteolin pretreatment (P > 0.05; Supplementary Figure 1M, https://links.lww.com/CM9/B188). Overall, these results indicate that miRNA-23a is involved in apoptosis during MI/RI, and that Tfpi2 is a target gene of miRNA-23a. We also used adenovirus containing miR-23a or Tfpi2 to explore the effects on apoptosis, infarct size, and left ventricular function in MI/RI models after luteolin pretreatment in vivo. The TFPI2 protein levels decreased in the MI/RI group (P < 0.001, vs. sham group) but increased in the luteolin + MI/RI group (P < 0.05, vs. MI/RI group). Compared with the MI/RI group, the TFPI2 protein levels decreased in the Ad-miR-23a + MI/RI, Ad-miR-23a + MI/RI + luteolin Ad-Tfpi2 shRNA + MI/RI, and Ad-Tfpi2 shRNA + MI/RI + luteolin groups (all P < 0.05), but increased in the Ad-miR-23a antago + MI/RI and Ad-miR-23a antago + MI/RI + luteolin groups (all P < 0.05; Supplementary Figure 4A, https://links.lww.com/CM9/B188). Western blotting analysis showed that Bax expression was upregulated in the Ad-miR-23a + MI/RI group whereas Bcl-2 expression was downregulated (P < 0.001 and P < 0.05, vs. the MI/RI group, respectively), but the opposite trend was observed in the Ad-miR-23a antago + MI/RI group. Compared with the MI/RI group, Bax and cleaved caspase-3 expression were upregulated in the Ad-Tfpi2 shRNA + MI/RI group, whereas Bcl-2 and caspase-3 expression were downregulated (all P < 0.05; Supplementary Figure 4B, https://links.lww.com/CM9/B188). MI/RI increased the myocardial infarct size (P < 0.001) but it decreased under luteolin pretreatment (P < 0.001, vs. MI/RI group). The myocardial infarct sizes were larger in the Ad-miR-23a + MI/RI and Ad-Tfpi2 shRNA + MI/RI groups (P < 0.01and P < 0.05, respectively), but smaller in the Ad-miR-23a antago + MI/RI group (P < 0.001, vs. MI/RI group; Supplementary Figure 1N, https://links.lww.com/CM9/B188). The hemodynamic indices comprising left ventricular systolic pressure, +dp/pt, and –dp/pt decreased after MI/RI injury, but these indices improved under luteolin pretreatment (all P < 0.05, vs. MI/RI group). Compared with the MI/RI group, these indices decreased in the Ad-miRNA-23a + MI/RI and Ad-Tfpi2 shRNA + MI/RI groups, but increased in the Ad-miRNA-23a antago + MI/RI group (all P < 0.05). The changes in left ventricular end-diastolic pressure exhibited the opposite trend [Supplementary Figure 4C, https://links.lww.com/CM9/B188]. In conclusion, this study identified the molecular mechanism responsible for the association between miRNA-23a and MI/RI. Luteolin pretreatment protected against MI/RI via miRNA-23a-mediated upregulation of TFPI2 both in vitro and in vivo. These findings further elucidate the mechanisms associated with cardiac I/R injury and the effective protection provided by luteolin. Funding This study was supported by grants from the National Natural Science Foundation of China (No. 81570326), Natural Science Foundation of Jiangsu Province (Nos. BK20141139, BK20190988), Jiangsu Commission of Health Foundation (No. H2018005), and Key Research and Development program of Xuzhou (No. KC20097). Conflicts of interest None.

Similarity and diversity of genetic architecture for complex traits between East Asian and European populations

Abstract: Abstract Background: Genome-wide association studies have detected a large number of single-nucleotide polymorphisms (SNPs) associated with complex traits in diverse ancestral groups. However, the trans-ethnic similarity and diversity of genetic architecture is not well understood currently. Results: By leveraging summary statistics of 37 traits from East Asian ( N max =254,373) or European ( N max =693,529) populations, we first evaluated the trans-ethnic genetic correlation ( ρ g ) and found substantial evidence of shared genetic overlap underlying these traits between the two populations, with ranging from 0.53 (se=0.11) for adult-onset asthma to 0.98 (se=0.17) for hemoglobin A1c. However, 88.9% of the genetic correlation estimates were significantly less than one, indicating potential heterogeneity in genetic effect across populations. We next identified common associated SNPs using the conjunction conditional false discovery rate method and observed 21.7% of trait-associated SNPs can be identified simultaneously in both populations. Among these shared associated SNPs, 20.8% showed heterogeneous influence on traits between the two ancestral populations. Moreover, we demonstrated that population-common associated SNPs often exhibited more consistent linkage disequilibrium and allele frequency pattern across ancestral groups compared to population-specific or null ones. We also revealed population-specific associated SNPs were much likely to undergo natural selection compared to population-common associated SNPs. Conclusions: Our study provides an in-depth understanding of similarity and diversity regarding genetic architecture for complex traits across diverse populations, and can assist in trans-ethnic association analysis, genetic risk prediction, and causal variant fine mapping.

Data from Multicenter Randomized Double-Blind Phase III Trial of Donafenib in Progressive Radioactive Iodine-Refractory Differentiated Thyroid Cancer

Abstract: <div>AbstractPurpose:<p>The phase II/III study of donafenib was initiated when there was no available treatment indicated for Chinese patients with progressive radioactive iodine-refractory differentiated thyroid cancer (RAIR-DTC). Donafenib, an oral tyrosine kinase inhibitor (TKI), showed good efficacy and tolerability in the phase II study. We aimed to further evaluate the antitumor activity and safety of donafenib in Chinese patients with RAIR-DTC.</p>Patients and Methods:<p>This multicenter, double-blind, placebo-controlled, phase III study enrolled 191 patients with progressive RAIR–DTC and randomized in a ratio of 2:1 to donafenib (300 mg twice daily, <i>n</i> = 128) or matched placebo (<i>n</i> = 63). An open-label donafenib treatment period was allowed upon disease progression. The primary endpoint was progression-free survival (PFS) assessed by the independent review committee. The second endpoints include objective response rate (ORR), disease control rate (DCR), safety, etc.</p>Results:<p>Donafenib demonstrated prolonged median PFS over placebo [12.9 vs. 6.4 months; hazard ratio (HR), 0.39; 95% confidence interval (CI), 0.25–0.61; <i>P</i> < 0.0001] in Chinese patients with RAIR–DTC. Improved ORR (23.3% vs. 1.7%; <i>P</i> = 0.0002) and DCR (93.3% vs. 79.3%; <i>P</i> = 0.0044) were observed in the donafenib group over placebo. For donafenib, the most common grade ≥ 3 treatment-related adverse events (AE) included hypertension (13.3%) and hand–foot syndrome (12.5%), 42.2% underwent dose reduction or interruption, and 6.3% experienced discontinuation.</p>Conclusions:<p>Donafenib was well-tolerated and demonstrated clinical benefit in terms of improved PFS, ORR, and DCR in patients with RAIR-DTC. The results suggest that donafenib could be a new treatment option for patients with RAIR-DTC.</p></div>

<scp>CircAGK</scp> regulates high dihydrotestosterone‐induced apoptosis in <scp>DPCs</scp> through the <scp>miR‐3180‐5p/BAX</scp> axis

Abstract: The incidence of androgen alopecia (AGA), also known as seborrheic alopecia, has surged in recent years, and onset is occurring at younger ages. Dermal papilla cells (DPCs) are key to maintaining hair cycling, and apoptosis-driven processes in DPCs are closely related to hair follicle regeneration. Circular RNAs (circRNAs) are widely present in the human body and are closely related to the occurrence and development of many diseases. Currently, the biological functions of circRNAs in AGA are largely unknown. Whole-transcriptome sequencing was used to screen differential circRNA expression profiles between AGA patients and non-AGA patients. We found that hsa_circ_0002980 (circAGK) was significantly highly expressed in the AGA group. CircAGK promoted DPC apoptosis in the presence of high dihydrotestosterone (DHT) (15 nmol/L). By regulating the miR-3180-5p/BAX axis, circAGK promotes DPC apoptosis in a high DHT environment in vitro and inhibits hair growth in AGA mice in vivo, indicating that circAGK is a potential target for the clinical treatment of AGA.

Inhibitory Effects of a Maleimide Compound on the Virulence Factors of <i>Candida albicans</i>.

Abstract: Candidiasis caused by Candida albicans infection has long been a serious human health problem. The pathogenicity of C. albicans is mainly due to its virulence factors, which are novel targets of antifungal drugs for low risk of resistance development. In this study, we identified a maleimide compound [1-(4-methoxyphenyl)-1hydro-pyrrole-2,5-dione, MPD] that exerts effective anti-virulence activity. It could inhibit the process of adhesion, filamentation, and biofilm formation in C. albicans. In addition, it exhibited low cytotoxicity, hemolytic activity, and drug resistance development. Moreover, in Galleria mellonella-C. albicans (in vivo) infection model, the survival time of infected larvae was significantly prolonged under the treatment of MPD. Further, mechanism research revealed that MPD increased farnesol secretion by upregulating the expression of Dpp3. The increased farnesol inhibited the activity of Cdc35, which then decreased the intracellular cAMP content resulting in the inhibition of virulence factors via the Ras1-cAMP-Efg1 pathway. In all, this study evaluated the inhibitory effect of MPD on various virulence factors of C. albicans and identified the underlying mechanisms. This suggests a potential application of MPD to overcome fungal infections in clinics.

The ubiquitin E3 ligase MDM2 induces chemoresistance in colorectal cancer by degradation of ING3

Abstract: Abstract Chemoresistance is an obstacle for colorectal cancer (CRC) treatment. This study investigates the role of the ubiquitin E3 ligase MDM2 in affecting cell growth and chemosensitivity in CRC cells by modifying the transcription factor inhibitor of growth protein 3 (ING3). The expression of MDM2 and ING3 in CRC tissues was predicted by bioinformatics analysis, followed by expression validation and their interaction in CRC HCT116 and LS180 cells. Ectopic overexpression or knockdown of MDM2/ING3 was performed to test their effect on proliferation and apotptosis as well as chemosensitivity of CRC cells. Finally, the effect of MDM2/ING3 expression on the in vivo tumorigenesis of CRC cells was examined through subcutaneous tumor xenograft experiment in nude mice. MDM2 promoted ubiquitin-proteasome pathway degradation of ING3 through ubiquitination and diminished its protein stability. Overexpression of MDM2 downregulated ING3 expression, which promoted CRC cell proliferation and inhibited the apoptosis. The enhancing role of MDM2 in tumorigenesis and resistance to chemotherapeutic drugs was also confirmed in vivo. Our findings highlight that MDM2 modifies the transcription factor ING3 by ubiquitination-proteasome pathway degradation, thus reducing ING3 protein stability, which finally promotes CRC cell growth and chemoresistance.

Principal neurons in the olfactory cortex mediate bidirectional modulation of seizures

Abstract: Although the piriform cortex (PC) has been previously implicated as a critical node for seizure generation and propagation, the underlying neural mechanism has remained unclear. Here, we found increased excitability in PC neurons during amygdala kindling acquisition. Optogenetic or chemogenetic activation of PC pyramidal neurons promoted kindling progression, whereas inhibition of these neurons retarded seizure activities induced by electrical kindling in the amygdala. Furthermore, chemogenetic inhibition of PC pyramidal neurons alleviated the severity of kainic acid-induced acute seizures. These results demonstrate that PC pyramidal neurons bidirectionally modulate seizures in temporal lobe epilepsy, providing evidence for the efficacy of PC pyramidal neurons as a potential therapeutic target for epileptogenesis. Key points While the piriform cortex (PC) is an important olfactory centre critically involved in olfactory processing and plays a crucial role in epilepsy due to its close connection with the limbic system, how the PC regulates epileptogenesis is largely unknown. In this study, we evaluated the neuronal activity and the role of pyramidal neurons in the PC in the mouse amygdala kindling model of epilepsy. PC pyramidal neurons are hyperexcited during epileptogenesis. Optogenetic and chemogenetic activation of PC pyramidal neurons significantly promoted seizures in the amygdala kindling model, whereas selective inhibition of these neurons produced an anti-epileptic effect for both electrical kindling and kainic acid-induced acute seizures. The results of the present study indicate that PC pyramidal neurons bidirectionally modulate seizure activity.

Data from Multicenter Randomized Double-Blind Phase III Trial of Donafenib in Progressive Radioactive Iodine-Refractory Differentiated Thyroid Cancer

Abstract: <div>AbstractPurpose:<p>The phase II/III study of donafenib was initiated when there was no available treatment indicated for Chinese patients with progressive radioactive iodine-refractory differentiated thyroid cancer (RAIR-DTC). Donafenib, an oral tyrosine kinase inhibitor (TKI), showed good efficacy and tolerability in the phase II study. We aimed to further evaluate the antitumor activity and safety of donafenib in Chinese patients with RAIR-DTC.</p>Patients and Methods:<p>This multicenter, double-blind, placebo-controlled, phase III study enrolled 191 patients with progressive RAIR–DTC and randomized in a ratio of 2:1 to donafenib (300 mg twice daily, <i>n</i> = 128) or matched placebo (<i>n</i> = 63). An open-label donafenib treatment period was allowed upon disease progression. The primary endpoint was progression-free survival (PFS) assessed by the independent review committee. The second endpoints include objective response rate (ORR), disease control rate (DCR), safety, etc.</p>Results:<p>Donafenib demonstrated prolonged median PFS over placebo [12.9 vs. 6.4 months; hazard ratio (HR), 0.39; 95% confidence interval (CI), 0.25–0.61; <i>P</i> < 0.0001] in Chinese patients with RAIR–DTC. Improved ORR (23.3% vs. 1.7%; <i>P</i> = 0.0002) and DCR (93.3% vs. 79.3%; <i>P</i> = 0.0044) were observed in the donafenib group over placebo. For donafenib, the most common grade ≥ 3 treatment-related adverse events (AE) included hypertension (13.3%) and hand–foot syndrome (12.5%), 42.2% underwent dose reduction or interruption, and 6.3% experienced discontinuation.</p>Conclusions:<p>Donafenib was well-tolerated and demonstrated clinical benefit in terms of improved PFS, ORR, and DCR in patients with RAIR-DTC. The results suggest that donafenib could be a new treatment option for patients with RAIR-DTC.</p></div>

Bioprinting EphrinB2-Modified Dental Pulp Stem Cells with Enhanced Osteogenic Capacity for Alveolar Bone Engineering

Abstract: Bioprinting, a technology that allows depositing living cells and biomaterials together into a complex tissue architecture with desired pattern, becomes a revolutionary technology for fabrication of engineered constructs. Previously, we have demonstrated that EphrinB2-modified dental pulp stem cells (DPSCs) are expected to be promising seed cells with enhanced osteogenic differentiation capability for alveolar bone regeneration. In this study, we aimed to bioprint EphrinB2-overexpressing DPSCs with low-concentrated Gelatin methacrylate (GelMA) hydrogels into three-dimensional (3D) constructs. The printability of GelMA (5% w/v) and the structural fidelity of bioprinted constructs were examined. Then, viability, proliferation, morphology, and osteogenic differentiation of DPSCs in bioprinted constructs were measured. Finally, the effect of EphrinB2 overexpression on osteogenic differentiation of DPSCs in bioprinted constructs was evaluated. Our results demonstrated that GelMA (5% w/v) in a physical gel form was successfully bioprinted into constructs with various shapes and patterns using optimized printing parameters. Embedded DPSCs showed round-like morphology, and had a high viability (91.93% ± 8.38%) and obvious proliferation (∼1.9-fold increase) 1 day after printing. They also showed excellent osteogenic potential in bioprinted constructs. In bioprinted 3D constructs, EphrinB2-overexpressing DPSCs expressed upregulated osteogenic markers, including ALP, BMP2, RUNX2, and SP7, and generated more mineralized nodules, as compared with Vector-DPSCs. Taken together, this study indicated that fabrication of bioprinted EphrinB2-DPSCs–laden constructs with enhanced osteogenic potential was possible, and 3D bioprinting strategy combined with EphrinB2 gene modification was a promising way to create bioengineered constructs for alveolar bone regeneration. In this work, we first optimized printing parameters of 5% Gelatin methacrylate (GelMA) and investigated stability of bioprinted constructs. Then, comparison of cell properties between bioprinted and casted hydrogel indicated that bioprinting had no obvious side effects on viability, proliferation, and osteogenic differentiation of dental pulp stem cells (DPSCs). Finally, bioprinted constructs loaded with EphrinB2-DPSCs exhibited enhanced osteogenic potential in vitro. In conclusion, our study reported optimized bioprinting parameters of low-concentrated GelMA, and provided a promising strategy for alveolar bone regeneration by combining bioprinting and EphrinB2 gene modification.