Anti-PD-L1 antibody retains antitumour effects while mitigating immunotherapy-related colitis in bladder cancer-bearing mice after CT-mediated intratumoral delivery.

Drug local delivery system that directly supply anti-cancer drugs to the tumor microenvironment (TME) results in excellent tumor control and minimizes side effects associated with the anti-cancer drugs. Immune checkpoint inhibitors (ICIs) have been the mainstay of cancer immunotherapy. However, the systemic administration of ICIs is accompanied by considerable immunotherapy-related toxicity. To explore whether an anti-PD-L1 antibody administered locally via a sustained-release gel-forming carrier retains its effective anticancer function while causing fewer colitis-like side effects, CT, a previously reported depot system, was used to locally deliver an anti-PD-L1 antibody together with curcumin to the TME in bladder cancer-bearing ulcerative colitis model mice. We showed that CT-mediated intratumoral coinjection of an anti-PD-L1 antibody and curcumin enabled sustained release of both the loaded anti-PD-L1 antibody and curcumin, which contributed to substantial anticancer effects with negligible side effects on the colons of the UC model mice. However, although the anti-PD-L1 antibody administered systemically synergized with the CT-mediated intratumoral delivery of curcumin in inhibiting tumour growth, colitis was significantly worsened by intraperitoneal administration of anti-PD-L1 antibody. These findings suggested that CT is a promising agent for the local delivery of anticancer drugs, as it can allow effective anticancer functions to be retained while sharply reducing the adverse side effects associated with the systemic administration of these drugs.

Synergistic anti-tumour activity of sorafenib in combination with pegylated resveratrol is mediated by Akt/mTOR/p70S6k-4EBP-1 and c-Raf7MEK/ERK signaling pathways.

Introduction: To investigate the inhibitory effect of sorafenib combined with PEGylated resveratrol on renal cell carcinoma (RCC) and its potential mechanism. Methods: MTT assay was used to detect the inhibitory effects of PEGylated resveratrol and sorafenib alone or combination on proliferation of RCC cells. Scratch and transwell assays were performed to examine the effects on the in vitro migration and invasion of RCC cells, respectively. The anti-tumor activity as well as splenic lymphocyte proliferation of the combination therapy was evaluated in the RCC xenograft mouse model. Western blotting method was used to detect changes in proteins involved in the antitumor efficacy related signaling pathways. Results: Inhibitory effects of PEGylated resveratrol combined with sorafenib incubation on the proliferation of Renca cells was synergistically enhanced compared with the mono-incubation group (both P < 0.01, CI < 1). Scratch and transwell assays revealed that combined incubation could significantly inhibit the migration and invasion of 786-O cells in vitro. Combined PEGylated resveratrol with sorafenib could significantly inhibit the growth of Renca renal carcinoma in mice with the tumor growth inhibition (TGI) of 85.5% and one achieved complete remission on D14, while the two monotherapies were both below 43% on D14, suggesting that current combination may have synergistic anti-renal carcinoma activity. Compared with the control group, PEGylated resveratrol combined with sorafenib in vivo promoted the proliferation of unactivated splenic lymphocytes and the proliferation of lymphocytes stimulated with concanavalin A and lipopolysaccharide. Western blotting results showed that combination therapy may suppress the growth of renal cell carcinoma by inhibiting AKT/mTOR/p70S6k-4EBP-1 and c-Raf7MEK/ERK signaling pathways. Conclusion: PEGylated resveratrol combined with sorafenib can achieve synergistic anti-RCC activity, and the mechanism may be related to the inhibition of Akt/mTOR/p70S6k-4EBP-1 and c-Raf7MEK/ERK signaling pathways.

GCKR and ADIPOQ gene polymorphisms in women with gestational diabetes mellitus.

AIMS: To investigate the associations of GCKR and ADIPOQ variants with the risk of gestational diabetes mellitus (GDM) in Chinese women. METHODS: GCKR rs1260326, ADIPOQ rs266729, and rs1501299 were selected and genotyped in 519 GDM patients and 498 controls. Candidate SNPs were genotyped using multiplex polymerase chain reaction (PCR) combined with next-generation sequencing methods, and the association of these SNPs with GDM was analyzed. RESULTS: We found that GCKR rs1260326 was significantly associated with an increased risk of GDM in the allele model, the codominant model (CC vs. TT), the dominant model, the recessive model, and the genotypic model distributions (p = 0.0029, p = 0.0022, p = 0.0402, p = 0.0038, and p = 0.0028, respectively). The rs1260326 polymorphism was shown to be associated with 1 h-OGTT level and gravidity in GDM patients (CC vs. TT: p = 0.0475 and p = 0.0220, respectively). Diastolic blood pressure (DBP) was significantly higher in the GDM patients with the rs266729 GG genotype compared to those with the CC or CG genotype (p = 0.0444 and p = 0.0339, respectively). The DBP of the GDM patients with the rs1501299 GT genotype was lower than that of those with the GG genotype (p = 0.0197). There was a weak linkage disequilibrium value between the GCKR and ADIPOQ SNPs. CONCLUSIONS: The genes GCKR and ADIPOQ may be involved in the pathophysiology of GDM.

RecON: Online learning for sensorless freehand 3D ultrasound reconstruction.

Sensorless freehand 3D ultrasound (US) reconstruction based on deep networks shows promising advantages, such as large field of view, relatively high resolution, low cost, and ease of use. However, existing methods mainly consider vanilla scan strategies with limited inter-frame variations. These methods thus are degraded on complex but routine scan sequences in clinics. In this context, we propose a novel online learning framework for freehand 3D US reconstruction under complex scan strategies with diverse scanning velocities and poses. First, we devise a motion-weighted training loss in training phase to regularize the scan variation frame-by-frame and better mitigate the negative effects of uneven inter-frame velocity. Second, we effectively drive online learning with local-to-global pseudo supervisions. It mines both the frame-level contextual consistency and the path-level similarity constraint to improve the inter-frame transformation estimation. We explore a global adversarial shape before transferring the latent anatomical prior as supervision. Third, we build a feasible differentiable reconstruction approximation to enable the end-to-end optimization of our online learning. Experimental results illustrate that our freehand 3D US reconstruction framework outperformed current methods on two large, simulated datasets and one real dataset. In addition, we applied the proposed framework to clinical scan videos to further validate its effectiveness and generalizability.

Unraveling the interaction mechanism between collagen and alcohols with different chain lengths and hydroxyl positions.

The present study systematically investigated the effects of alcohols, including methanol, ethanol, n-butanol, and propanol with different hydroxyl group numbers and locations on the thermal stability and molecular aggregation behavior of collagen. The results of ultra-sensitive differential scanning calorimetry (US-DSC), dynamic light scattering (DLS) and intrinsic fluorescence showed that with the increase of carbon chain length, alcohols can denature collagen, accompanied by transition in triple helical structure, promoted aggregation behavior, and altered molecular interactions. However, with the number of hydroxyl groups in alcohol molecules increased, the thermal stability of collagen increased and the molecules tended to disperse. Furthermore, radial distribution function (RDF) results showed that alcohols can change the structure of the hydration layer around collagen, thus altering the aggregation morphology of collagen molecules in solution. The results of the interaction between components in different alcohol systems demonstrated that with the decrease of alcohol polarity, bridge bond networks were formed between collagen molecules. Specifically, it was found that because the hydroxyl groups in 1,3-propanediol are located at both ends of the carbon chain, the reticular bridge bond structure formed between the collagen molecules changed into chain-like bridge structure. The bridge bonds between collagen molecules were considered to be weak cross-linking, which was an important reason for the destruction of collagen structure. In this study, the mechanism of interaction between different alcohols and collagen was elucidated, which will be helpful for further development of complex alcohol and collagen products.

Unraveling the Role of Hydroxyproline in Maintaining the Thermal Stability of the Collagen Triple Helix Structure Using Simulation.

The thermal stability of collagen has an important effect on its practical applications. Many believe that hydroxyproline (Hyp) improves the structural stability of collagen molecules. In this study, for the first time, a method of building natural collagen molecular models was described. We constructed a collagen model with typical triple-helix structure and calculated the hydrogen bond energy between collagen α chains. The calculated hydrogen bond energy was consistent with the experimental results of differential scanning calorimetry. After the calculation simulation, we verified that the hydrogen bond energy between collagen chains was positively correlated with Hyp content in the models and an increased Hyp content in the model was beneficial in improving the thermal resistance of the structure. In addition, we found that thermal unfolding did not occur simultaneously along the entire molecule but started in the regions with less Hyp content. This study provides a collagen model with a natural collagen amino acid sequence, which will be helpful for further investigation of the physical and chemical properties of natural collagen.

Changes in aggregation behavior of collagen molecules in solution with varying concentrations of acetic acid.

A critical aggregation concentration of 0.30-0.50mg/mL was previously obtained for type I collagen at 0.1M acetic acid (AA). In the present study, the aggregation behavior of collagen in solution (0.5mg/mL) in the presence of 0.1-2.0M AA was investigated. Circular dichroism showed that the three helix structure was maintained across the whole AA concentration range. However, the ratio of positive peak intensity over negative peak intensity varied depending on the conformational state of collagen aggregates. Ultra-sensitive differential scanning calorimetry revealed that transition temperatures Tm1 and Tm2 decreased by 8.35°C and 7.80°C, respectively, between 0.1M and 2.0M, indicating a possible relationship between the aggregation state and the thermal effect. The surrounding polarity of collagen molecules in solution containing pyrene was investigated by fluorescence spectroscopy, which demonstrated that disaggregation of collagen aggregates was enhanced with increasing AA concentration. This observation was correlated with changes in collagen fiber size observed by atomic force microscopy. Furthermore, collagen tyrosine residues were blue-shifted in an intrinsic fluorescence spectra, further indicating changes in aggregation behavior with increasing AA concentration. Finally, the dynamic response of collagen molecules to AA was analyzed by two-dimensional correlation fluorescence spectra.