Novel method for preparing fish collagen gels with excellent physicochemical properties via the dehydration of ethanol

Content: Fish collagen has been considered to be an alternative for mammalian collagen, however, physicochemical properties of fish collagen-based materials such as gels are so far not adequate for actual application. In the present study, we prepared two types of fish collagen gels with sufficient elasticity: i) dehydrated fibrillogenesis collagen gels (DFCG), which were fabricated via collagen self-assembly followed by immersion in different concentrations of ethanol solutions, and ii) dehydrated cross-linking collagen gels (DCCG), which were fabricated via collagen self-assembly and simultaneous cross-linking followed by immersion in ethanol solution. Furthermore, the physicochemical properties of DFCG and DCCG were analyzed by atomic force microscopy, differential scanning calorimetry, enzymatic degradation and dynamic viscoelastic measurements. The microstructure of DFCG was consisted of characteristic Dperiodic collagen fibrils and insusceptible of ethanol concentrations (20-100% (v/v)). However, the thermal stability, remaining weight after enzymatic degradation and mechanical properties of DFCG distinctly increased with the increase of ethanol dose, possiblely ascribing that ethanol with higher polarity might dehydrate partial free water of DFCG and strengthen the interactions of hydrogen bond. Especially, for the gel treated by 100% (v/v) enthanol, Td increased by 32.7 °C and G′ was 55-folds than those of undehydrated gel (43.1 °C and 239.2 Pa). In the case of DCCG, the formation of collagen fibrils was depended on the concentrations of N-hydroxysuccinimide adipic acid derivative (NHS-AA), which was converted to [NHS-AA]/[NH2] ratios (calculated by the [active ester group] of NHS-AA and [ε-NH2] of lysine and hydroxylysine residues of collagen). As the ratio= 0.05, the characteristic D-periodic fibrils were still formed and the treatment of 60% (v/v) ethanol increased the Td (52.5 °C) and G′ (7388 Pa) values of the gel compared with those of uncross-linked gel (49 °C and 2064.32 Pa, respectively), majorly resulting from the effects of covalent cross-linking bonds and hydrogen bonds. However, when the ratio= 0.2, the collagen self-assembly was intensively inhibited and the dehydration of free water within gel structure in the absence of thick fibrils led to the shrinkage of the gel and an obvious decrease in Td (42 °C) and G′ (432 Pa). Although the [NHS-AA]/[NH2] ratio further increased to 0.8, the thermal stability and elasticity of the gel enhanced mildly suggesting that the presence of thick fibrils formed via the self-assembly was significantly crucial for reinforcing the gels. Take-Away: The fish collagen gels with excellent elasticity were prepared via the treatment of ethanol. The physicochemical properties of the dehydrated gels were depended on the concentrations of ethanol. The presence of characteristic D-periodic fibrils was significantly crucial for reinforcing the gels

Activation of Protein Serine/Threonine Phosphatase PP2Cα Efficiently Prevents Liver Fibrosis

Over-activation of TGFβ signaling pathway and uncontrolled cell proliferation of hepatic stellate cells (HSCs) play pivotal roles in liver fibrogenesis, while the protein serine/threonine phosphatase PP2Cα was reported to negatively regulate TGFβ signaling pathway and cell cycle. Our study aimed to investigate the role of PP2Cα in liver fibrogenesis.The effects of PP2Cα activation on liver fibrosis were investigated in human HSCs and primary rat HSCs in vitro using western blotting, real-time PCR, nuclear translocation, cell viability and cell cycle analyses. The antifibrogenic effects in carbon tetrachloride (CCl(4))- and bile duct ligation (BDL)-induced mice in vivo were assessed using biochemical, histological and immunohistochemical analyses. The results demonstrated that activation of PP2Cα by overexpression or the new discovered small molecular activator NPLC0393 terminated TGFβ-Smad3 and TGFβ-p38 signaling pathways, induced cell cycle arrest in HSCs and decreased α-smooth muscle actin (α-SMA) expression, collagen deposition and hepatic hydroxyproline (HYP) level in CCl(4)- and BDL-induced mice.Our findings suggested that PP2Cα activation might be an attractive new strategy for treating liver fibrosis while the small molecular activator NPLC0393 might represent a lead compound for antifibrogenic drug development. Moreover, our study might provide the first evidence for the role of PP2C family members in the fibrotic disease

Splicing factor TRA2A contributes to esophageal cancer progression via a noncanonical role in lncRNA m⁶A methylation

Transformer 2 alpha homolog (TRA2A), a member of the serine/arginine-rich splicing factor family, has been shown to control mRNA splicing in development and cancers. However, it remains unclear whether TRA2A is involved in lncRNA regulation. In the present study, we found that TRA2A was upregulated and correlated with poor prognosis in esophageal cancer. Downregulation of TRA2A suppressed the tumor growth in xenograft nude mice. Epitranscriptomic microarray showed that depletion of TRA2A affected global lncRNA methylation similarly to the key m6A methyltransferase, METTL3, by silencing. MeRIP-qPCR, RNA pull-down, CLIP analyses, and stability assays indicated that ablation of TRA2A reduced m6A-modification of the oncogenic lncRNA MALAT1, thus inducing structural alterations and reduced stability. Furthermore, Co-IP experiments showed TRA2A directly interacted with METTL3 and RBMX, which also affected the writer KIAA1429 expression. Knockdown of TRA2A inhibited cell proliferation in a manner restored by RBMX/KIAA1429 overexpression. Clinically, MALAT1, RBMX, and KIAA1429 were prognostic factors of worse survival in ESCA patients. Structural similarity-based virtual screening in FDA-approved drugs repurposed nebivolol, a β1-adrenergic receptor antagonist, as a potent compound to suppress the proliferation of esophageal cancer cells. Cellular thermal shift and RIP assay indicated that nebivolol may compete with MALAT1 to bind TRA2A. In conclusion, our study revealed the noncanonical function of TRA2A, which coordinates with multiple methylation proteins to promote oncogenic MALAT1 during ESCA carcinogenesis.</p

OmniQuant: Omnidirectionally Calibrated Quantization for Large Language Models

Large language models (LLMs) have revolutionized natural language processing tasks. However, their practical deployment is hindered by their immense memory and computation requirements. Although recent post-training quantization (PTQ) methods are effective in reducing memory footprint and improving the computational efficiency of LLM, they hand-craft quantization parameters, which leads to low performance and fails to deal with extremely low-bit quantization. To tackle this issue, we introduce an Omnidirectionally calibrated Quantization (OmniQuant) technique for LLMs, which achieves good performance in diverse quantization settings while maintaining the computational efficiency of PTQ by efficiently optimizing various quantization parameters. OmniQuant comprises two innovative components including Learnable Weight Clipping (LWC) and Learnable Equivalent Transformation (LET). LWC modulates the extreme values of weights by optimizing the clipping threshold. Meanwhile, LET tackles activation outliers by shifting the challenge of quantization from activations to weights through a learnable equivalent transformation. Operating within a differentiable framework using block-wise error minimization, OmniQuant can optimize the quantization process efficiently for both weight-only and weight-activation quantization. For instance, the LLaMA-2 model family with the size of 7-70B can be processed with OmniQuant on a single A100-40G GPU within 1-16 hours using 128 samples. Extensive experiments validate OmniQuant's superior performance across diverse quantization configurations such as W4A4, W6A6, W4A16, W3A16, and W2A16. Additionally, OmniQuant demonstrates effectiveness in instruction-tuned models and delivers notable improvements in inference speed and memory reduction on real devices. Codes and models are available at \url{https://github.com/OpenGVLab/OmniQuant}.Comment: Updated result with 2-bit quantization. A differentiable quantization method for LL

A HIV-1 heterosexual transmission chain in Guangzhou, China: a molecular epidemiological study

<p>Abstract</p> <p>Background</p> <p>We conducted molecular analyses to confirm four clustering HIV-1 infections (Patient A, B, C & D) in Guangzhou, China. These cases were identified by epidemiological investigation and suspected to acquire the infection through a common heterosexual transmission chain.</p> <p>Methods</p> <p><it>Env C2V3V4 </it>region, <it>gag p17/p24 </it>junction and partial <it>pol </it>gene of HIV-1 genome from serum specimens of these infected cases were amplified by reverse transcription polymerase chain reaction (RT-PCR) and nucleotide sequenced.</p> <p>Results</p> <p>Phylogenetic analyses indicated that their viral nucleotide sequences were significantly clustered together (bootstrap value is 99%, 98% and 100% in <it>env</it>, <it>gag </it>and <it>pol </it>tree respectively). Evolutionary distance analysis indicated that their genetic diversities of <it>env</it>, <it>gag </it>and <it>pol </it>genes were significantly lower than non-clustered controls, as measured by unpaired <it>t</it>-test (<it>env </it>gene comparison: <it>p </it>< 0.005; <it>gag </it>gene comparison: <it>p </it>< 0.005; <it>pol </it>gene comparison: <it>p </it>< 0.005).</p> <p>Conclusion</p> <p>Epidemiological results and molecular analyses consistently illustrated these four cases represented a transmission chain which dispersed in the locality through heterosexual contact involving commercial sex worker.</p

Continuous and Noninvasive Measurement of Arterial Pulse Pressure and Pressure Waveform using an Image-free Ultrasound System

The local beat-to-beat local pulse pressure (PP) and blood pressure waveform of arteries, especially central arteries, are important indicators of the course of cardiovascular diseases (CVDs). Nevertheless, noninvasive measurement of them remains a challenge in the clinic. This work presents a three-element image-free ultrasound system with a low-computational method for real-time measurement of local pulse wave velocity (PWV) and diameter waveforms, enabling real-time and noninvasive continuous PP and blood pressure waveforms measurement without calibration. The developed system has been well-validated in vitro and in vivo. In in vitro cardiovascular phantom experiments, the results demonstrated high accuracy in the measurement of PP (error < 3 mmHg) and blood pressure waveform (root-mean-square-errors (RMSE) < 2 mmHg, correlation coefficient (r) > textgreater 0.99). In subsequent human carotid experiments, the system was compared with an arterial tonometer, which showed excellent PP accuracy (mean absolute error (MAE) = 3.7 +- 3.4 mmHg) and pressure waveform similarity (RMSE = 3.7 +- 1.6 mmHg, r = 0.98 +- 0.01). Furthermore, comparative experiments with the volume clamp device demonstrated the system's ability to accurately trace blood pressure changes (induced by deep breathing) over a period of one minute, with the MAE of DBP, MAP, and SBP within 5 +- 8 mmHg. The present results demonstrate the accuracy and reliability of the developed system for continuous and noninvasive measurement of arterial PP and blood pressure waveform measurements, with potential applications in the diagnosis and prevention of CVDs.Comment: 13 pages, 12 figure

Intestinal fungi contribute to development of alcoholic liver disease

This study was supported in part by NIH grants R01 AA020703, U01 AA021856 and by Award Number I01BX002213 from the Biomedical Laboratory Research & Development Service of the VA Office of Research and Development (to B.S.). K.H. was supported by a DFG (Deutsche Forschungsgemeinschaft) fellowship (HO/ 5690/1-1). S.B. was supported by a grant from the Swiss National Science Foundation (P2SKP3_158649). G.G. received funding from the Yale Liver Center NIH P30 DK34989 and R.B. from NIAAA grant U01 AA021908. A.K. received support from NIH grants RC2 AA019405, R01 AA020216 and R01 AA023417. G.D.B. is supported by funds from the Wellcome Trust. We acknowledge the Human Tissue and Cell Research (HTCR) Foundation for making human tissue available for research and Hepacult GmbH (Munich, Germany) for providing primary human hepatocytes for in vitro analyses. We thank Dr. Chien-Yu Lin Department of Medicine, Fu-Jen Catholic University, Taiwan for statistical analysis.Peer reviewedPublisher PD

Optimization method for prefabricated restroom envelope energy saving characteristics in hot summer and cold winter zone

In order to reduce the restroom envelope energy consumption, one optimization method on basis of analyzing the influence of heat transfer coefficient on the performance of a prefabricated restroom envelope in a hot summer and cold winter zone was proposed. An energy-consuming model of prefabricated restroom in Nanjing is initially built based on Designer's Simulation Toolkit software. Subsequently, the effect of external walls, rooftops, external windows with various thermal characteristics on the building envelope is analyzed respectively. Simultaneously, a method that only changes the heat transfer coefficient of the prefabricated restroom envelope while keeping other parameters unchanged is adopted. Results show that, for a prefabricated restroom, the optimal range of heat transfer coefficient of the external wall, rooftop, and external window in hot summer and cold winter zone is 0.199∼0.22, 0.16∼0.19, and 3.0∼3.1 W/(m 2 ·K), respectively. When the window-to-wall ratio is less than 0.2, the priority of the wall heat transfer coefficient on building energy consumption is higher than that of the rooftop heat transfer coefficient, simultaneously, the rooftop heat transfer coefficient has priority higher than window heat transfer coefficient. Thus, it is of great significance to optimize the design of the prefabricated restroom envelope in a hot summer and cold winter zone, which provides relative reference for thermal performance improvement of prefabricated restrooms

miR-26a inhibits atherosclerosis progression by targeting TRPC3

Abstract Background Atherosclerosis, a chronic multi-factorial vascular disease, has become a predominant cause of a variety of cardiovascular disorders. miR-26a was previously reported to be involved in atherosclerosis progression. However, the underlying mechanism of miR-26a in atherosclerosis remains to be further explained. Methods High-fat diet (HFD)-fed apolipoprotein E (apoE)−/− mice and oxidized low-density lipoprotein (ox-LDL)-stimulated human aortic endothelial cells (HAECs) were established as in vivo and in vitro models of atherosclerosis. RT-qPCR and western blot analysis were performed to measure the expression of miR-26a and transient receptor potential canonical 3 (TRPC3), respectively. Binding between miR-26a and TRPC3 was predicted with bioinformatics software and verified using a dual luciferase reporter assay. The effects of miR-26a on the lipid accumulation, atherosclerotic lesion, and inflammatory response in HFD-fed apoE−/− mice were investigated by a colorimetric enzymatic assay system, hematoxylin–eosin and oil-Red-O staining, and ELISA, respectively. Additionally, the effects of miR-26a or combined with TRPC3 on cell viability, apoptosis and the nuclear factor-kappa B (NF-κB) pathway in ox-LDL-stimulated HAECs were evaluated by MTT assay, TUNEL assay, and western blot, respectively. Results miR-26a was downregulated in HFD-fed apoE−/− mice and ox-LDL-stimulated HAECs. miR-26a overexpression inhibited the pathogenesis of atherosclerosis by attenuating hyperlipidemia, atherosclerotic lesion and suppressing inflammatory response in HFD-fed apoE−/− mice. Moreover, miR-26a overexpression suppressed inflammatory response and the NF-κB pathway, promoted cell viability and inhibited apoptosis in ox-LDL-stimulated HAECs. Additionally, TRPC3 was demonstrated to be a direct target of miR-26a. Enforced expression of TRPC3 reversed the effects of miR-26a on cell viability, apoptosis, and the NF-κB pathway in ox-LDL-treated HAECs. Conclusions miR-26a alleviated the development of atherosclerosis by regulating TRPC3, providing a potential target for atherosclerosis treatment

Novel method for preparing fish collagen gels with excellent physicochemical properties via the dehydration of ethanol

Content: Fish collagen has been considered to be an alternative for mammalian collagen, however, physicochemical properties of fish collagen-based materials such as gels are so far not adequate for actual application. In the present study, we prepared two types of fish collagen gels with sufficient elasticity: i) dehydrated fibrillogenesis collagen gels (DFCG), which were fabricated via collagen self-assembly followed by immersion in different concentrations of ethanol solutions, and ii) dehydrated cross-linking collagen gels (DCCG), which were fabricated via collagen self-assembly and simultaneous cross-linking followed by immersion in ethanol solution. Furthermore, the physicochemical properties of DFCG and DCCG were analyzed by atomic force microscopy, differential scanning calorimetry, enzymatic degradation and dynamic viscoelastic measurements. The microstructure of DFCG was consisted of characteristic Dperiodic collagen fibrils and insusceptible of ethanol concentrations (20-100% (v/v)). However, the thermal stability, remaining weight after enzymatic degradation and mechanical properties of DFCG distinctly increased with the increase of ethanol dose, possiblely ascribing that ethanol with higher polarity might dehydrate partial free water of DFCG and strengthen the interactions of hydrogen bond. Especially, for the gel treated by 100% (v/v) enthanol, Td increased by 32.7 °C and G′ was 55-folds than those of undehydrated gel (43.1 °C and 239.2 Pa). In the case of DCCG, the formation of collagen fibrils was depended on the concentrations of N-hydroxysuccinimide adipic acid derivative (NHS-AA), which was converted to [NHS-AA]/[NH2] ratios (calculated by the [active ester group] of NHS-AA and [ε-NH2] of lysine and hydroxylysine residues of collagen). As the ratio= 0.05, the characteristic D-periodic fibrils were still formed and the treatment of 60% (v/v) ethanol increased the Td (52.5 °C) and G′ (7388 Pa) values of the gel compared with those of uncross-linked gel (49 °C and 2064.32 Pa, respectively), majorly resulting from the effects of covalent cross-linking bonds and hydrogen bonds. However, when the ratio= 0.2, the collagen self-assembly was intensively inhibited and the dehydration of free water within gel structure in the absence of thick fibrils led to the shrinkage of the gel and an obvious decrease in Td (42 °C) and G′ (432 Pa). Although the [NHS-AA]/[NH2] ratio further increased to 0.8, the thermal stability and elasticity of the gel enhanced mildly suggesting that the presence of thick fibrils formed via the self-assembly was significantly crucial for reinforcing the gels. Take-Away: The fish collagen gels with excellent elasticity were prepared via the treatment of ethanol. The physicochemical properties of the dehydrated gels were depended on the concentrations of ethanol. The presence of characteristic D-periodic fibrils was significantly crucial for reinforcing the gels