Protocol to screen for Sorafenib resistance regulators using pooled lentiviral shRNA library and a Sorafenib-resistant hepatocellular carcinoma cell model.

Approaches to study therapy resistance in HCC (hepatocellular carcinoma) are limited, especially when using HCC models in vitro. Here, we present a protocol to establish an in vitro Sorafenib-resistant human HCC cell model and conduct an shRNA-mediated synthetic lethal screen in established Sorafenib-resistant HCC cell lines to identify critical regulators of Sorafenib resistance. We describe steps for RNA sequencing and functional analysis to reveal the mode of action of potential candidates in conferring therapy resistance to HCC cells. For complete details on the use and execution of this protocol, please refer to Gao et al. (2021a)1 and Gao et al. (2021b).2.

The Effect of Cooling Rates on Thermal, Crystallization, Mechanical and Barrier Properties of Rotational Molding Polyamide 11 as the Liner Material for High-Capacity High-Pressure Vessels.

The rapid development of hydrogen fuel cells has been paralleled by increased demand for lightweight type IV hydrogen storage vessels with high hydrogen storage density, which raises the performance requirements of internal plastic liners. An appropriate manufacturing process is important to improve the quality of polymer liners. In this paper, DSC, WAXD, a universal testing machine and a differential pressure gas permeameter were used to investigate the effect of the cooling rate of the rotational molding polyamide 11 on the thermal, crystallization, mechanical and barrier properties. The cooling rate is formulated according to the cooling rate that can be achieved in actual production. The results suggest that two PA11 liner materials initially exhibited two-dimensional (circular) growth under non-isothermal crystallization conditions and shifted to one-dimensional space growth due to spherulite collision and crowding during the secondary crystallization stage. The slower the cooling process, the greater the crystallinity of the specimen. The increase in crystallinity significantly improved the barrier properties of the two PA11 liner materials, and the gas permeability coefficient was 2-3-fold higher than at low crystallinity. Moreover, the tensile strength, the tensile modulus, the flexural strength, and the flexural modulus increased, and the elongation at break decreased as the crystallinity increased.

Optimization of the Laminate Structure of a Composite Cylinder Based on the Combination of Response Surface Methodology (RSM) and Finite Element Analysis (FEA).

This study optimized the laminate structure of a composite cylinder under the constraint of minimum layup thickness. Based on the progressive damage theory, a finite element model of the cylinder was established, and the NOL ring tensile test was used to verify the accuracy of the damage theory. The winding angle, the number of layers, and the helical/hoop ratio (the stacking sequence) were selected as the optimization factors, and the burst pressure value was used to evaluate the quality of the laminate structure. Then the orthogonal experiments were designed by RSM. Combined with FEA, the function model of the burst pressure of the gas cylinder and each optimization factor was established to obtain the optimal layering scheme, satisfying the minimum burst pressure. In addition, finite element analysis was used to verify the optimal scheme, demonstrating that the error of the burst pressure predicted by the quadratic model established by the response surface design was not more than 5%. This study provides a faster and more efficient optimization method for the optimization design of composite gas cylinder layers.

Anatomic resection versus radiofrequency ablation with an ablative margin ≥ 1.0 cm for solitary small hepatocellular carcinoma measuring ≤ 3 cm: Comparison of long-term outcomes using propensity score matching analysis.

PURPOSE: To compare the long-term outcomes of anatomic resection (AR) and radiofrequency ablation (RFA) with an ablative margin (AM) of ≥ 1.0 cm as first-line treatment for solitary hepatocellular carcinoma measuring ≤ 3 cm. METHODS: Two hundred and fifty-one patients who underwent AR (n = 156) or RFA (ablative margin ≥ 1.0 cm, n = 95) at any of 6 tertiary hospitals from 2009 to 2018 were enrolled. Propensity score matched analysis (PSM) were used to compare overall survival (OS), recurrence-free survival (RFS), and perioperative outcomes. Univariate and multivariate analyses were performed to identify prognostic factors associated with RFS and OS. RESULTS: PSM created 67 patient-pairs. After 96 months of follow-up, RFA with an ablative margin ≥ 1.0 cm and AR showed comparable 1-year, 3-year, 5-year, and 8-year OS rates before (P = 0.580) and after (P = 0.640) PSM. However, RFS was better at 1, 3, 5, and 8 years after AR before (P = 0.0036) and after (P = 0.017) PSM. The operation time and postoperative hospital stay were significantly longer in the AR group than in the RFA group before and after PSM (P < 0.05). Multivariate analysis identified age and type of treatment to be independent prognostic factors for RFS and age and hepatitis C to be associated with OS. CONCLUSIONS: Long-term OS was not significantly different between AR and RFA with an AM ≥ 1.0 cm in patients with a solitary hepatocellular carcinoma measuring ≤ 3 cm; but, RFS appeared to be better after AR than after RFA. However, RFA was associated with fewer perioperative complications and a shorter postoperative hospital stay.

Synergistic Treatment of Congo Red Dye with Heat Treated Low Rank Coal and Micro-Nano Bubbles.

In this study, the adsorption method and micro-nano bubble (MNB) technology were combined to improve the efficiency of organic pollutant removal from dye wastewater. The adsorption properties of Congo red (CR) on raw coal and semi-coke (SC) with and without MNBs were studied. The mesoporosity of the coal strongly increased after the heat treatment, which was conducive to the adsorption of macromolecular organics, such as CR, and the specific surface area increased greatly from 2.787 m2/g to 80.512 m2/g. MNBs could improve the adsorption of both raw coal and SC under different pH levels, temperatures and dosages. With the use of MNBs, the adsorption capacity of SC reached 169.49 mg/g, which was much larger than that of the raw coal at 15.75 mg/g. The MNBs effectively reduced the adsorption time from 240 to 20 min. In addition, the MNBs could ensure the adsorbent maintained a good adsorption effect across a wide pH range. The removal rate was above 90% in an acidic environment and above 70% in an alkaline environment. MBs can effectively improve the rate of adsorption of pollutants by adsorbents. SC was obtained from low-rank coal through a rapid one-step heating treatment and was used as a kind of cheap adsorbent. The method is thus simple and easy to implement in the industrial context and has the potential for industrial promotion.

Safety and efficacy of microwave versus radiofrequency ablation for large hepatic hemangioma: a multicenter retrospective study with propensity score matching.

OBJECTIVES: To compare the safety and efficacy of microwave ablation (MWA) and radiofrequency ablation (RFA) for such hemangiomas (5-9.9 cm in diameter). METHODS: This multicenter retrospective cohort study investigated the differences in technical success, ablation time, complete ablation, complications, hospital stay, and clinical response between MWA and RFA. A total of 452 patients with hepatic hemangiomas were screened. Propensity score matching was performed. Univariable and multivariate regression analyses were used. RESULTS: Among the 452 patients, 394 met the eligibility criteria and completed the follow-up. After the propensity score matching analysis, 72 pairs of patients were created. No technical failures were found. The RFA group had a longer ablation time (48.63 ± 18.11 min versus [vs.] 37.18 ± 15.86 min, p < 0.001), higher morbidity of hemoglobinuria (77.78% vs. 50.00%, p < 0.001), and longer hospital stay (5.01 ± 1.56 days vs. 4.34 ± 1.42 days, p < 0.05) than the MWA group. The treatment methods (p = 0.032, OR = 0.105, 95% CI = 0.013-0.821), size of the hemangioma (p = 0.021, OR = 5.243, 95% CI = 1.285-21.391), and time of ablation (p = 0.031, OR = 1.145, 95% CI = 1.013-1.294) were significant independent risk factors associated with hemoglobinuria. No recurrence or delayed complications were observed. There were no differences in complete ablation, clinical response, and health-related quality of life between the groups. CONCLUSIONS: MWA and RFA appear to be effective treatments for large hepatic hemangiomas. However, MWA had a shorter ablation time than RFA, and MWA was associated with fewer hemolysis-related complications and shorter hospital stays. KEY POINTS: • MWA and RFA appear to be effective treatments for large hepatic hemangiomas. • MWA had a shorter ablation time than RFA. • MWA was associated with fewer hemolysis-related complications and shorter hospital stays.

Deep Reinforcement Learning for Solving the Heterogeneous Capacitated Vehicle Routing Problem.

Existing deep reinforcement learning (DRL)-based methods for solving the capacitated vehicle routing problem (CVRP) intrinsically cope with a homogeneous vehicle fleet, in which the fleet is assumed as repetitions of a single vehicle. Hence, their key to construct a solution solely lies in the selection of the next node (customer) to visit excluding the selection of vehicle. However, vehicles in real-world scenarios are likely to be heterogeneous with different characteristics that affect their capacity (or travel speed), rendering existing DRL methods less effective. In this article, we tackle heterogeneous CVRP (HCVRP), where vehicles are mainly characterized by different capacities. We consider both min-max and min-sum objectives for HCVRP, which aim to minimize the longest or total travel time of the vehicle(s) in the fleet. To solve those problems, we propose a DRL method based on the attention mechanism with a vehicle selection decoder accounting for the heterogeneous fleet constraint and a node selection decoder accounting for the route construction, which learns to construct a solution by automatically selecting both a vehicle and a node for this vehicle at each step. Experimental results based on randomly generated instances show that, with desirable generalization to various problem sizes, our method outperforms the state-of-the-art DRL method and most of the conventional heuristics, and also delivers competitive performance against the state-of-the-art heuristic method, that is, slack induction by string removal. In addition, the results of extended experiments demonstrate that our method is also able to solve CVRPLib instances with satisfactory performance.

Safety and effectiveness of laparoscopic intratumoral resection facilitated by coagulation of giant hepatic hemangioma: a matched case-control study and literature review.

BACKGROUND: To report the safety and effectiveness of laparoscopic intratumoral resection facilitated by coagulation (LIRC) compared with laparoscopic hepatectomy (LH) in treating giant hepatic hemangioma. METHODS: From 2017 to 2020, 19 consecutive patients with giant hepatic hemangioma (≥ 10 cm) received LIRC in one center. We selected a subgroup of 103 patients treated by LH in other four centers who well matched the 19 consecutive patients treated with LIRC, in a 1:1 fashion based on the tumor location, tumor size, and body mass index. Furthermore, the differences in technical success, operative time, operative blood loss, change of laboratory indexes, hospital stays, complication and clinical responds are compared between the two groups. RESULTS: Technical success was achieved in all 38 patients. Patients in the LIRC group had a relative shorter operative time (P < 0.001) and less operative blood loss (P = 0.003). The serum levels of C-reactive protein (CRP), total bilirubin (TBil), alanine aminotransferase (ALT), and aspartate transaminase (AST) were elevated significantly (P < 0.05) 1 day after the resection and returned to normal within 7 days in both groups; however, relatively lower serum levels of those indexes were observed in the LIRC group (P < 0.05). The total complication rate was relatively lower in the LIRC group compared with the LH group (P = 0.029). Patients in the LIRC group had shorter hospital stays than those in the LH group (P = 0.010). The clinical response was similar in the two groups. CONCLUSIONS: LIRC is safe and effective for treating giant hepatic hemangioma.

YAP/TAZ and ATF4 drive resistance to Sorafenib in hepatocellular carcinoma by preventing ferroptosis.

Understanding the mechanisms underlying evasive resistance in cancer is an unmet medical need to improve the efficacy of current therapies. In this study, a combination of shRNA-mediated synthetic lethality screening and transcriptomic analysis revealed the transcription factors YAP/TAZ as key drivers of Sorafenib resistance in hepatocellular carcinoma (HCC) by repressing Sorafenib-induced ferroptosis. Mechanistically, in a TEAD-dependent manner, YAP/TAZ induce the expression of SLC7A11, a key transporter maintaining intracellular glutathione homeostasis, thus enabling HCC cells to overcome Sorafenib-induced ferroptosis. At the same time, YAP/TAZ sustain the protein stability, nuclear localization, and transcriptional activity of ATF4 which in turn cooperates to induce SLC7A11 expression. Our study uncovers a critical role of YAP/TAZ in the repression of ferroptosis and thus in the establishment of Sorafenib resistance in HCC, highlighting YAP/TAZ-based rewiring strategies as potential approaches to overcome HCC therapy resistance.

Complications of Radiofrequency Ablation for Hepatic Hemangioma: A Multicenter Retrospective Analysis on 291 Cases.

PURPOSE: To report the complications of radiofrequency ablation (RFA) for hepatic hemangioma. PATIENTS AND METHODS: Investigators from six centers performed RFA for hepatic hemangioma and used a standardized follow-up protocol. Data were collected from 291 patients, including 253 patients with hepatic hemangioma 5 to 9.9 cm in diameter (group A) and 38 with hepatic hemangioma ≥ 10 cm (group B). Technical success, complete ablation, and complications attributed to the RFA procedure were reported. Analysis of variance was used to determine whether the major complication rate was related to tumor size or clinical experience. RESULTS: A total of 304 lesions were treated in 291 patients. Technical success was achieved without adverse events in all cases. A total of 301 lesions were completely ablated, including 265 of 265 (100%) lesions in group A, and 36 of 39 (92.31%) in group B. The rate of technology-related complications was similar in groups A and B (5.14% (13/253) and 13.16% (5/38), respectively; P = 0.121). Moreover, all technology-related complications occurred during the early learning curve period. The rate of hemolysis-related complications in two groups were 83.40% (211/253) and 100% (38/38) (P =0.007) and the systemic inflammatory response syndrome-related complications in two groups were 33.99% (86/253) and 86.84% (33/38) (P<0.001). There were no delayed complications in either group. CONCLUSION: RFA is minimally invasive, safe, and effective for hepatic hemangiomas 5 to 9.9 cm in diameter. More clinical data are needed to confirm the safety of RFA for hepatic hemangiomas ≥ 10 cm.

USP29-mediated HIF1α stabilization is associated with Sorafenib resistance of hepatocellular carcinoma cells by upregulating glycolysis.

Understanding the mechanisms underlying evasive resistance in cancer is an unmet medical need to improve the efficacy of current therapies. In hepatocellular carcinoma (HCC), aberrant expression of hypoxia-inducible factor 1 α (HIF1α) and increased aerobic glycolysis metabolism are drivers of resistance to therapy with the multi-kinase inhibitor Sorafenib. However, it has remained unknown how HIF1α is activated and how its activity and the subsequent induction of aerobic glycolysis promote Sorafenib resistance in HCC. Here, we report the ubiquitin-specific peptidase USP29 as a new regulator of HIF1α and of aerobic glycolysis during the development of Sorafenib resistance in HCC. In particular, we identified USP29 as a critical deubiquitylase (DUB) of HIF1α, which directly deubiquitylates and stabilizes HIF1α and, thus, promotes its transcriptional activity. Among the transcriptional targets of HIF1α is the gene encoding hexokinase 2 (HK2), a key enzyme of the glycolytic pathway. The absence of USP29, and thus of HIF1α transcriptional activity, reduces the levels of aerobic glycolysis and restores sensitivity to Sorafenib in Sorafenib-resistant HCC cells in vitro and in xenograft transplantation mouse models in vivo. Notably, the absence of USP29 and high HK2 expression levels correlate with the response of HCC patients to Sorafenib therapy. Together, the data demonstrate that, as a DUB of HIF1α, USP29 promotes Sorafenib resistance in HCC cells, in parts by upregulating glycolysis, thereby opening new avenues for therapeutically targeting Sorafenib-resistant HCC in patients.

Different physiological responses of C3 and C4 plants to nanomaterials.

Several studies have previously reported that nanomaterial uptake and toxicity in plants are species dependent. However, the differences between photosynthetic pathways, C3 and C4, following nanomaterial exposure are poorly understood. In the current work, wheat and rice, two C3 pathway species are compared to amaranth and maize, which utilize the C4 photosynthetic mechanism. These plants were cultured in soils which were spiked with CuO, Ag, TiO2, MWCNT, and FLG nanomaterials. Overall, the C4 plant exhibited higher resilience to NM stress than C3 plants. In particular, significant differences were observed in chlorophyll contents with rice returning a 40.9-54.2% decrease compared to 3.5-15.1% for maize. Fv/Fm levels were significantly reduced by up to 51% in rice whereas no significant reductions were observed in amaranth and maize. Furthermore, NM uptake in the C3 species was greater than that in C4 plants, a trend that was also seen in metal concentration. TEM results showed that CuO NPs altered the chloroplast thylakoid structure in rice leaves and a large number of CuO NPs were observed in the vascular sheath cells. In contrast, there were no significant changes in the chloroplasts in the vascular sheath and no significant CuO NPs were found in maize leaves. This study was the first to systematically characterize the effect of metal and carbon-based nanomaterials in soil on C3 and C4 plants, providing a new perspective for understanding the impact of nanomaterials on plants.

Effect of micro-H2O and micro-O2 on the decomposition characteristics of insulating medium C3F7CN gas using molecular dynamics and transition state method.

In recent years, heptafluoroisobutyronitrile (C3F7CN) has been proved to be a potential eco-friendly insulating medium to replace sulfur hexafluoride (SF6, the strong greenhouse gas). In this paper, the effect of micro-H2O and micro-O2 on the decomposition of C3F7CN was investigated based on the reactive force field molecular dynamics (ReaxFF-MD) and transition state theory (TST). It was found that H2O obviously promoted the decomposition of C3F7CN, and new products HF, COF2, CO, and NO were generated. The influence of O2 on the C3F7CN dissociation was weaker than that of H2O, and O2 slightly promoted the C3F7CN decomposition only when 50 O2 molecules were added. The simultaneous presence of H2O and O2 promoted the decomposition of C3F7CN, the promotion of which was closed to H2O existing alone. The calculation results showed that the energy barriers of the two reactions forming COF2 were 31.38 kcal/mol and 23.85 kcal/mol, respectively, which indicated that the reactions were difficult to proceed spontaneously. The energy barrier of F + H2O → HF + OH was relatively lower than that of COF2. These values corresponded well to the ReaxFF simulation results. This study provides theoretical support for the effect of H2O and O2 on the decomposition of C3F7CN.

LATS1 but not LATS2 represses autophagy by a kinase-independent scaffold function.

Autophagy perturbation represents an emerging therapeutic strategy in cancer. Although LATS1 and LATS2 kinases, core components of the mammalian Hippo pathway, have been shown to exert tumor suppressive activities, here we report a pro-survival role of LATS1 but not LATS2 in hepatocellular carcinoma (HCC) cells. Specifically, LATS1 restricts lethal autophagy in HCC cells induced by sorafenib, the standard of care for advanced HCC patients. Notably, autophagy regulation by LATS1 is independent of its kinase activity. Instead, LATS1 stabilizes the autophagy core-machinery component Beclin-1 by promoting K27-linked ubiquitination at lysine residues K32 and K263 on Beclin-1. Consequently, ubiquitination of Beclin-1 negatively regulates autophagy by promoting inactive dimer formation of Beclin-1. Our study highlights a functional diversity between LATS1 and LATS2, and uncovers a scaffolding role of LATS1 in mediating a cross-talk between the Hippo signaling pathway and autophagy.

SnS2 /Co3 S4 Hollow Nanocubes Anchored on S-Doped Graphene for Ultrafast and Stable Na-Ion Storage.

SnS2 has been widely studied as an anode material for sodium-ion batteries (SIBs) based on the high theoretical capacity and layered structure. Unfortunately, rapid capacity decay associated with volume variation during cycling limits practical application. Herein, SnS2 /Co3 S4 hollow nanocubes anchored on S-doped graphene are synthesized for the first time via coprecipitation and hydrothermal methods. When applied as the anode for SIBs, the sample delivers a distinguished charge specific capacity of 1141.8 mAh g-1 and there is no significant capacity decay (0.1 A g-1 for 50 cycles). When the rate is increased to 0.5 A g-1 , it presents 845.7 mAh g-1 after cycling 100 times. Furthermore, the composite also exhibits an ultrafast sodium storage capability where 392.9 mAh g-1 can be obtained at 10 A g-1 and the charging time is less than 3 min. The outstanding electrochemical properties can be ascribed to the enhancement of conductivity for the addition of S-doped graphene and the existence of p-n junctions in the SnS2 /Co3 S4 heterostructure. Moreover, the presence of mesopores between nanosheets can alleviate volume expansion during cycling as well as being beneficial for the migration of Na+ .

The SIAH2-NRF1 axis spatially regulates tumor microenvironment remodeling for tumor progression.

The interactions between tumor cells with their microenvironments, including hypoxia, acidosis and immune cells, lead to the tumor heterogeneity which promotes tumor progression. Here, we show that SIAH2-NRF1 axis remodels tumor microenvironment through regulating tumor mitochondrial function, tumor-associated macrophages (TAMs) polarization and cell death for tumor maintenance and progression. Mechanistically, low mitochondrial gene expression in breast cancers is associated with a poor clinical outcome. The hypoxia-activated E3 ligase SIAH2 spatially downregulates nuclear-encoded mitochondrial gene expression including pyruvate dehydrogenase beta via degrading NRF1 (Nuclear Respiratory Factor 1) through ubiquitination on lysine 230, resulting in enhanced Warburg effect, metabolic reprogramming and pro-tumor immune response. Dampening NRF1 degradation under hypoxia not only impairs the polarization of TAMs, but also promotes tumor cells to become more susceptible to apoptosis in a FADD-dependent fashion, resulting in secondary necrosis due to the impairment of efferocytosis. These data represent that inhibition of NRF1 degradation is a potential therapeutic strategy against cancer.