Broad-spectrum kinome profiling identifies CDK6 upregulation as a driver of lenvatinib resistance in hepatocellular carcinoma.

Increasing evidence has demonstrated that drug resistance can be acquired in cancer cells by kinase rewiring, which is an obstacle for efficient cancer therapy. However, it is technically challenging to measure the expression of protein kinases on large scale due to their dynamic range in human proteome. We employ a lysine-targeted sulfonyl fluoride probe, named XO44, which binds to 133 endogenous kinases in intact lenvatinib-resistant hepatocellular carcinoma (HCC) cells. This analysis reveals cyclin-dependent kinase 6 (CDK6) upregulation, which is mediated by ERK/YAP1 signaling cascade. Functional analyses show that CDK6 is crucial in regulation of acquired lenvatinib resistance in HCC via augmentation of liver cancer stem cells with clinical significance. We identify a noncanonical pathway of CDK6 in which it binds and regulates the activity of GSK3ß, leading to activation of Wnt/ß-catenin signaling. Consistently, CDK6 inhibition by palbociclib or degradation by proteolysis targeting chimeras (PROTACs) is highly synergistic with lenvatinib in vitro. Interestingly, palbociclib not only exerts maximal growth suppressive effect with lenvatinib in lenvatinib-resistant HCC models but also reshapes the tumor immune microenvironment. Together, we unveil CDK6 as a druggable target in lenvatinib-resistant HCC and highlight the use of a chemical biology approach to understand nongenetic resistance mechanisms in cancer.

Long-term effectiveness of group-based diabetes self-management on glycosylated haemoglobin for people with type 2 diabetes in community: a protocol of systematic review and meta-analysis.

INTRODUCTION: The rapid rise in the prevalence of diabetes has a negative impact on patients' quality of life. Diabetes self-management group education is cost-effective and efficient for patients to control blood glucose. However, there are no consistent standards for self-management group education, and its long-term effects (≥12 months) are unclear. Although a few systematic reviews evaluated the long-term effects, they did not make clear provisions on the content of self-management, and the number and sample size of included studies were small, which may lead to misclassification bias and reporting bias. Therefore, we plan to conduct this systematic review to evaluate the long-term effects of self-management group education and determine the effects of different self-management characteristics on glycosylated haemoglobin (HbA1c). METHODS AND ANALYSIS: We will retrieve Chinese databases (Wanfang, Chinese Hospital Knowledge Warehouse) and English databases (PubMed, ScienceDirect, EMBASE, Web of Science, Bailian Platform, Cochrane Central Register of Controlled Trials, Google Scholar) for randomly controlled trials and cluster randomly controlled trials of which participants are adults with type 2 diabetes mellitus. We will manually search citation lists and trial registries, and consult authors to obtain relevant articles. The retrieval time range will be from the establishment of the database to July 2020 to avoid omitting relevant studies. The primary outcome will be HbA1c. The secondary outcomes will be fasting plasma glucose, postprandial blood glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, systolic blood pressure, diastolic blood pressure, body mass index, waist circumference and death event. Two reviewers will independently conduct article screening and assessment of risk of bias, with a third reviewer arbitrating if necessary. We will give priority to the use of meta-analysis to evaluate the pooled effects of all outcomes. For the outcomes of unrecognised sources of heterogeneity, missing data and less than three related studies, narrative synthesis approach will be used. ETHICS AND DISSEMINATION: Ethical approval is not required for this systematic review. We plan to present the findings in a peer-reviewed scientific journal, relevant and responsible organisations, and training meetings. PROSPERO REGISTRATION NUMBER: CRD42020209011.

Anaerobic digestion of elephant grass hydrolysate: Biogas production, substrate metabolism and outlet effluent treatment.

Elephant grass (Pennisetum purpureum) acid hydrolysate was used as substrate for anaerobic digestion for the first time. Within short period (ten days), the organic materials (sugars and organic acids) in the elephant grass hydrolysate could be utilized efficiently for stable biogas production that the COD removal, biogas yield, and CH4 content were 91.3 ±â€¯2.0%, 0.561 ±â€¯0.014 m3/kg COD consumption, and 68.1 ±â€¯4.6%, respectively throughout this bioprocess. During anaerobic digestion, almost no volatile fatty acids (VFAs) was accumulated (merely <0.1 g/L acetic acid was found) and the outlet pH was very stable (7.3 ±â€¯0.1). Meanwhile, the furans including furfural and 5-hydroxymethylfurfural (HMF) existing in the inlet substrate could be degraded. After anaerobic digestion, the outlet effluent was treated by combination of Fe-C micro-electrolysis and Fenton reaction to remove 93.1% of residual COD and 98.6% of color. Considering the performance, cost, operation, and environmental influence, this technology is suitable for industrial treatment of waste elephant grass.

Difference of trace element exposed routes and their health risks between agriculture and pastoral areas in Bay County Xinjiang, China.

The concentration level related to toxicities of trace elements in drinking water, rice, wheat flour, and their associated negative impacts on human health have become an emergent issue in China. Because Xinjiang is the largest province in China with the majority of arable pasture land available for cultivation, it is important to analyze the concentrations of trace elements in relation to their toxicities in water, rice, and wheat flour and to investigate the health risk differences between agricultural and pastoral areas in Bay County, Xinjiang. The study results showed that (1) metal concentrations from drinking water, rice, and wheat flour were within the permissible limits; (2) concentration levels of trace elements and their total risk from drinking water and rice were higher in the agricultural areas than those in the pastoral areas, whereas concentration levels of trace elements and their total risk from wheat flour were higher in the pastoral areas than those in the agricultural areas; (3) the concentration level of the trace elements in rice were higher than in the wheat flour, but the risk from the wheat flour was higher than the risk from rice; (4) total non-cancer risk from the flour (HIf) in both areas exceeded the respective safe reference doses; (5) total cancer risk from the wheat flour, rice, and water exceeded the safety limit (1 × 10-4); (6) for the exposed population, arsenic was suggested as the most evident pollutant leading to carcinogenic concerns regarding the water, rice, and wheat flour; (7) the risk index from the wheat flour made up the highest percentage both in the total cancer risk and the non-cancer risk, followed by rice and then water; and (8) the human health risk was attributed to influence from the local environment in the agriculture areas, while it was attributed to the external environment in the pastoral areas. Graphical abstract.

Middle hepatic vein reconstruction in adult right lobe living donor liver transplantation improves recipient survival.

BACKGROUND: The efficacy and necessity of middle hepatic vein (MHV) reconstruction in adult-to-adult right lobe living donor liver transplantation (LDLT) remain controversial. The present study aimed to evaluate the survival beneficiary of MHV reconstructions in LDLT. METHODS: We compared the clinical outcomes of liver recipients with MHV reconstruction (n = 101) and without MHV reconstruction (n = 43) who underwent LDLT using right lobe grafts at our institution from January 2006 to May 2017. RESULTS: The overall survival (OS) rate of recipients with MHV reconstruction was significantly higher than that of those without MHV reconstruction in liver transplantation (P = 0.022; 5-yr OS: 76.2% vs 58.1%). The survival of two segments (segments 5 and 8) hepatic vein reconstruction was better than that of the only one segment (segment 5 or segment 8) hepatic vein reconstruction (P = 0.034; 5-yr OS: 83.6% vs 67.4%). The survival of using two straight vascular reconstructions was better than that using Y-shaped vascular reconstruction in liver transplantation with two segments hepatic vein reconstruction (P = 0.020; 5-yr OS: 100% vs 75.0%). The multivariate analysis demonstrated that MHV tributary reconstructions were an independent beneficiary prognostic factor for OS (hazard ratio=0.519, 95% CI: 0.282-0.954, P = 0.035). Biliary complications were significantly increased in recipients with MHV reconstruction (28.7% vs 11.6%, P = 0.027). CONCLUSIONS: MHV reconstruction ensured excellent outflow drainage and favored recipient outcome. The MHV tributaries (segments 5 and 8) should be reconstructed as much as possible to enlarge the hepatic vein anastomosis and reduce congestion.

Bacterial cellulose based superabsorbent production: A promising example for high value-added utilization of clay and biology resources.

Superabsorbent was synthesized from bacterial cellulose (BC) generated by in situ fermentation on bentonite inorganic gel (BIG). For BIG preparation, the effect of sodium agent's type and content, temperature and time of sodium-modification, and gelling agent's type and content on the viscosity of BIG were learned to optimize the synthesis process. For polymerization, the effect of different factors including ratio of monomer to substrate (modified BC from in situ fermentation), content of initiator and crosslinker, monomer neutralization degree, reaction temperature and time on the performance of composite (superabsorbent) synthesized were analyzed. Under optimal condition, the composite showed good water absorption, salts absorption, and water retention capacity. The original bentonite, sodium-based bentonite, BIG and composite structure was characterized by X-ray fluorescence (XRF), nuclear magnetic resonance (NMR), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM) and thermogravimetric analysis (TGA), and the characterization partly explained the performance of water absorption and thermal stability of the composite. Overall, this study provides one method for superabsorbent synthesis from low-cost and natural resources.

Fast Startup of Semi-Pilot-Scale Anaerobic Digestion of Food Waste Acid Hydrolysate for Biogas Production.

In this study, a fast startup of semi-pilot-scale anaerobic digestion of food waste acid hydrolysate for biogas production was carried out for the first time. During the period of fast startup, more than 85% of chemical oxygen demand (COD) can be degraded, and even more than 90% of COD can be degraded during the later stage of anaerobic digestion. During this anaerobic digestion process, the biogas yield, the methane yield, and the CH4 content in biogas were 0.542 ± 0.056 m3/kgCOD consumption, 0.442 ± 0.053 m3/kgCOD consumption, and 81.52 ± 3.05%, respectively, and these values were high and stable. Besides, the fermentation pH was very stable, in which no acidification was observed during the anaerobic digestion process (outlet pH was 7.26 ± 0.05 for the whole anaerobic digestion). Overall, the startup of this anaerobic digestion can be completed in a short period (the system can be stable 2 days after the substrate was pumped into the bioreactor), and anaerobic digestion of food waste acid hydrolysate is feasible and attractive for industrial treatment of food waste and biogas production.

Lumping kinetics of ABE fermentation wastewater treatment by oleaginous yeast Trichosporon cutaneum.

Lumping kinetics models were built for the biological treatment of acetone-butanol-ethanol (ABE) fermentation wastewater by oleaginous yeast Trichosporon cutaneum with different fermentation temperatures. Compared with high temperature (33°C, 306 K) and low temperature (23°C, 296 K), medium temperature (28°C, 301 K) was beneficial for the cell growth and chemical oxygen demand (COD) degradation during the early stage of fermentation but the final yeast biomass and COD removal were influenced little. By lumping method, the materials in the bioconversion network were divided into five lumps (COD, lipid, polysaccharide, other intracellular products, other extracellular products), and the nine rate constants (k1-k9) for the models can well explain the bioconversion laws. The Gibbs free energy (G) for this bioconversion was positive, showing that it cannot happen spontaneous, but the existence of yeast can after the chemical equilibrium and make the bioconversion to be possible. Overall, the possibility of using lumping kinetics for elucidating the laws of materials conversion in the biological treatment of ABE fermentation wastewater by T. cutaneum has been initially proved and this method has great potential for further application.

Using wastewater after lipid fermentation as substrate for bacterial cellulose production by Gluconacetobacter xylinus.

In this study, lipid fermentation wastewater (fermentation broth after separation with yeast biomass) with high Chemical Oxygen Demand (COD) value of 25,591 mg/L was used as substrate for bacterial cellulose (BC) production by Gluconacetobacter xylinus for the first time. After 5 days of fermentation, the highest BC yield (0.659 g/L) was obtained. Both monosaccharide and polysaccharides present in lipid fermentation wastewater could be utilized by G. xylinus simultaneously during fermentation. By this bioconversion, 30.0% of COD could be removed after 10 days of fermentation and the remaining wastewater could be used for further BC fermentation. The crystallinity of BC samples in lipid fermentation wastewater increased gradually during fermentation but overall the environment of lipid fermentation wastewater showed small influence on BC structure by comparison with that in traditional HS medium by using FE-SEM, FTIR, and XRD. By this work, the possibility of using lipid fermentation wastewater containing low value carbohydrate polymer (extracellular polysaccharides) for high value carbohydrate polymer (BC) production was proven.

Bacterial cellulose production from the litchi extract by Gluconacetobacter xylinus.

Although litchi has both nutrient and edible value, the extremely short preservation time limited its further market promotion. To explore processed litchi products with longer preservation time, litchi extract was selected as an alternative feedstock for production of bacterial cellulose (BC). After 2 weeks of static fermentation, 2.53 g/L of the BC membrane was obtained. The trace elements including magnesium (Mg) and sodium (Na) in the litchi extract were partly absorbed in the BC membrane, but no potassium (K) element was detected in it, curiously. Scanning electron microscope (SEM) photographs exhibited an ultrafine network nanostructure for the BC produced in the litchi extract. Analysis of the fourier-transform infrared spectroscopy (FTIR) confirmed the pellicles to be a cellulosic material. Interestingly, X-ray diffraction (XRD) results showed the BC membrane obtained from litchi extract had higher crystallinity of 94.0% than that from HS medium. Overall, the work showed the potential of producing high value-added polymer from litchi resources.

Utilization of corncob acid hydrolysate for bacterial cellulose production by Gluconacetobacter xylinus.

In this study, corncob acid hydrolysate was used as a substrate for bacterial cellulose (BC) production by Gluconacetobacter xylinus. After 2 weeks' static fermentation, a BC yield of 4 g/L could be obtained. Both effects of medium composition and fermentation condition on the BC production were evaluated. Most extra substrates (carbon and nitrogen sources) except mannitol, butyric acid, and levulinic acid showed no effect on the improvement of BC yield. Fermentation condition including fermentation mode, inoculation concentration, and initial pH showed certain influence on the BC yield and thus should be well controlled. The analysis by field emission scanning electron microscope (FE-SEM), Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) showed that the BC sample had obvious nano-network structure, clear functional groups that were found in cellulose, and relatively high crystallinity and crystallinity index value. Moreover, the BC sample had great water-holding capacity. Overall, corncob acid hydrolysate could be one promising substrate for BC production.

Beneficial Effect of Acetic Acid on the Xylose Utilization and Bacterial Cellulose Production by Gluconacetobacter xylinus.

In this work, acetic acid was found as one promising substrate to improve xylose utilization by Gluconacetobacter xylinus CH001. Also, with the help of adding acetic acid into medium, the bacterial cellulose (BC) production by G. xylinus was increased significantly. In the medium containing 3 g l(-1) acetic acid, the optimal xylose concentration for BC production was 20 g l(-1). In the medium containing 20 g l(-1) xylose, the xylose utilization and BC production by G. xylinus were stimulated by acetic acid within certain concentration. The highest BC yield (1.35 ± 0.06 g l(-1)) was obtained in the medium containing 20 g l(-1) xylose and 3 g l(-1) acetic acid after 14 days. This value was 6.17-fold higher than the yield (0.21 ± 0.01 g l(-1)) in the medium only containing 20 g l(-1) xylose. The results analyzed by FE-SEM, FTIR, and XRD showed that acetic acid affected little on the microscopic morphology and physicochemical characteristics of BC. Base on the phenomenon observed, lignocellulosic acid hydrolysates (xylose and acetic acid are main carbon sources present in it) could be considered as one potential substrate for BC production.

ZIP4, a novel determinant of tumor invasion in hepatocellular carcinoma, contributes to tumor recurrence after liver transplantation.

BACKGROUND AND PURPOSE: Recently, evidence that Zinc transporter ZRT/IRT-like protein 4 (ZIP4) is involved in invasiveness and apoptosis has emerged in pancreatic cancer and prostate cancer. Our aim was to assess the role of ZIP4 in invasiveness, migration and apoptosis of hepatocellular carcinoma (HCC). The prognostic value of ZIP4 in HCC after liver transplantation was evaluated. METHODS: The role of ZIP4 in HCC was investigated by overexpressing ZIP4 in BEL7402 and HepG2 cells and inhibiting ZIP4 in HuH-7 and HepG2 cells, using overexpression and shRNA plasmids in vitro studies. Immunohistochemical analysis was used to evaluate ZIP4 expression in HCC tissues from 60 patients undergoing liver transplantation, 36 cirrhotic tissue samples, and 6 normal tissue samples. Prognostic significance was assessed using the Kaplan-Meier method and the log-rank test. RESULTS: Specific suppression of ZIP4 reduced cell migration and invasiveness, whereas ZIP4 overexpression caused increases in cell migration and invasiveness. Furthermore, overexpression of ZIP4 resulted in increased expression of pro-metastatic genes (MMP-2, MMP-9) and decreased expression of pro-apoptotic genes (caspase-3, caspase-9, Bax). In contrast, suppression of ZIP4 resulted in an opposite effect. ZIP4 was more highly expressed in tumor tissues than non-tumor tissues (P < 0.0001). ZIP4 expression was significantly associated with tumor recurrence (P = 0.002), tumor node metastasis stage (P = 0.044), Child-Turcotte-Pugh score (P = 0.042), and tumor size (P = 0.022). Univariate analysis showed that ZIP4 expression was significantly associated with overall survival (P = 0.020) and tumor-free survival (P = 0.049). Multivariate analysis revealed that ZIP4 was an independent predictor of overall survival (P = 0.037) after liver transplantation. CONCLUSIONS: ZIP4 could promote migration, invasiveness, and suppress apoptosis in hepatocellular carcinoma, and represent a novel predictor of poor prognosis and therapeutic target for patients with HCC who undergo liver transplantation.

FOXC1 contributes to microvascular invasion in primary hepatocellular carcinoma via regulating epithelial-mesenchymal transition.

The existence of microvascular invasion (MVI) formation is one of the most important risk factors predicting poor outcome in hepatocellular carcinoma (HCC) and its mechanism remains largely unknown. Epithelial-Mesenchymal Transition (EMT) has been suggested to be involved in many steps of the invasion-metastasis cascade. To elucidate the possible contribution of EMT to MVI, we initially evaluated the expression of 8 EMT-related transcription factors (TFs) in HCC patients with or without MVI and found that FOXC1 expression was significantly higher in patients with MVI than those without MVI (P < 0.05). Knockdown of FOXC1 expression in HCC cells resulted in a partial conversion of their EMT progresses, mainly regulating the mesenchymal component. Ectopic expression of snail, twist or TGF-ß1 could induce expression of FOXC1, but none of the expression of snail, twist, slug or TGF-ß was consistently down-regulated in response to FOXC1 silencing, suggesting FOXC1 might operate the downstream of other EMT regulators. In addition, knockdown of FOXC1 expression led to cytoskeleton modification accompanied by decreased ability of cell proliferation, migration, and invasion. Meanwhile, some matrix metalloproteinases (MMPs) and VEGF-A were also simultaneously down-regulated. Together, our findings demonstrate that FOXC1 is one of candidate predictive markers of MVI, and that inhibition of FOXC1 expression can partially reverse EMT program, offering a potential molecular therapeutic target for reducing tumor metastasis in HCC patients.

Major influence of renal function on hyperlipidemia after living donor liver transplantation.

AIM: To investigate the impact of renal and graft function on post-transplant hyperlipidemia (PTHL) in living donor liver transplantation (LDLT). METHODS: A total of 115 adult patients undergoing LDLT from January 2007 to May 2009 at a single center were enrolled. Data were collected and analyzed by the China Liver Transplant Registry retrospectively. PTHL was defined as serum triglycerides ≥ 150 mg/dL or serum cholesterol ≥ 200 mg/dL or the need for pharmacologic treatment at the sixth month after LDLT. Early renal dysfunction (ERD) was defined as serum creatinine ≥ 2 mg/dL and/or the need for renal replacement therapy in the first post-transplant week. RESULTS: In 115 eligible patients, the incidence of PTHL was 24.3%. Recipients with PTHL showed a higher incidence of post-transplant cardiovascular events compared to those without PTHL (17.9% vs 4.6%, P = 0.037). Serum creatinine showed significant positive correlations with total serum triglycerides, both at post-transplant month 1 and 3 (P < 0.01). Patients with ERD had much higher pre-transplant serum creatinine levels (P < 0.001) and longer duration of pre-transplant renal insufficiency (P < 0.001) than those without ERD. Pre-transplant serum creatinine, graft-to-recipient weight ratio, graft volume/standard liver volume ratio, body mass index (BMI) and ERD were identified as risk factors for PTHL by univariate analysis. Furthermore, ERD [odds ratio (OR) = 9.593, P < 0.001] and BMI (OR = 6.358, P = 0.002) were identified as independent risk factors for PTHL by multivariate analysis. CONCLUSION: Renal function is closely associated with the development of PTHL in LDLT. Post-transplant renal dysfunction, which mainly results from pre-transplant renal insufficiency, contributes to PTHL.