Clinical-Radiological Characteristic for Predicting Ultra-Early Recurrence After Liver Resection in Solitary Hepatocellular Carcinoma Patients.

Objective: This study aims to identify independent risk factors for ultra-early recurrence in patients with early solitary hepatocellular carcinoma (HCC) and develop an individualized predictive nomogram for ultra-early recurrence. Materials and Methods: A total of 332 patients with early solitary HCC who underwent curative liver resection at our hospital from January 2015 to May 2021 were included in this study. Based on the patients' recurrence status at 6 months, they were divided into the non-ultra-early recurrence group and the ultra-early recurrence group. Univariate and multivariate Cox regression analyses were used to construct the nomogram, and internal validation of its performance was performed using calibration plots with bootstrapping. Results: Among the 332 patients with early solitary HCC, 39 (11.7%) experienced ultra-early recurrence. Tumor morphology, age > 46 years, AFP > 332.4 ng/mL, GGT > 51.2 U/L, ALP > 126 U/L, PT > 12.8 s, and satellite nodules were identified as independent prognostic factors for ultra-early recurrence in patients with early solitary HCC and were incorporated into the final predictive nomogram. The C-index of the nomogram and bootstrap resampling were 0.842 and 0.815, respectively. The calibration plot demonstrated good agreement between the predicted and observed probabilities of ultra-early recurrence, and DCA indicated the favorable clinical utility of the nomogram. Additionally, AFP > 332.4 ng/mL, AST > 35 U/L, GGT > 51.2 U/L, ALP > 126 U/L, tumor morphology, tumor size, satellite nodules, and intratumoral hemorrhage were identified as risk factors for overall survival in patients with early solitary HCC. Conclusion: Our study establishes a nomogram for predicting the postoperative ultra-early recurrence status in patients with early solitary HCC, which provides valuable supplementary decision-making information for clinical decision-makers and guides the selection of the most appropriate treatment strategy.

Source apportionment and migration characteristics of heavy metal(loid)s in soil and groundwater of contaminated site.

The rapid industrial growth has generated heavy metal(loid)s contamination in the soil, which poses a serious threat to the ecology and human health. In this study, 580 samples were collected in Henan Province, China, for source apportionment, migration characterization and health risk evaluation using self-organizing map, positive matrix factorization and multivariate risk assessment methods. The results showed that samples were classified into four groups and pollution sources included chromium slag dump, soil parent rock and abandoned factory. The contents of Cr, Pb, As and Hg were low in Group 1. Group 2 was characterized by total Cr, Cr(Ⅵ) and pH. The enrichment of total Cr and Cr(Ⅵ) in soil was mainly attributed to chromium slag dump, accounting for more than 84.0%. Group 3 was dominated by Hg and Pb. Hg and Pb were primarily attributed to abandoned factory, accounting for 84.7% and 70.0%, respectively. Group 4 was characterized by As. The occurrence of As was not limited to one individual region. The contribution of soil parent rock reached 83.0%. Furthermore, the vertical migration of As, Hg, Pb and Cr(Ⅵ) in soil was mainly influenced by medium permeability, pH and organic matter content. The trends of As, Pb, and Hg with depth were basically consistent with the trends of organic matter with depth, and were negatively correlated with the change in pH with depth. The trends of Cr(Ⅵ) with depth were basically consistent with the changes in pH with the depth. The content of Cr(Ⅵ) in the deep soil did not exceed the detection limits and Cr(Ⅵ) contamination occurred in the deep aquifer, suggesting that Cr(Ⅵ) in the deep groundwater originated from the leakage of shallow groundwater. The assessment indicated that the non-carcinogenic and carcinogenic risks for children and adults could not be neglected. Moreover, children were more susceptible than adults.

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

PURPOSE: 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. PATIENTS AND METHODS: 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, n = 128) or matched placebo (n = 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. RESULTS: 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; P < 0.0001] in Chinese patients with RAIR-DTC. Improved ORR (23.3% vs. 1.7%; P = 0.0002) and DCR (93.3% vs. 79.3%; P = 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. CONCLUSIONS: 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.

IR780-doped cobalt ferrite nanoparticles@poly(ethylene glycol) microgels as dual-enzyme immobilized micro-systems: Preparations, photothermal-responsive dual-enzyme release, and highly efficient recycling.

To solve the problems of separating dual enzymes from the carriers of dual-enzyme immobilized micro-systems and greatly increase the carriers' recycling times, photothermal-responsive micro-systems of IR780-doped cobalt ferrite nanoparticles@poly(ethylene glycol) microgels (CFNPs-IR780@MGs) are prepared. A novel two-step recycling strategy is proposed based on the CFNPs-IR780@MGs. First, the dual enzymes and the carriers are separated from the reaction system as a whole via magnetic separation. Second, the dual enzymes and the carriers are separated through photothermal-responsive dual-enzyme release so that the carriers can be reused. Results show that CFNPs-IR780@MGs is 281.4 ± 9.6 nm with a shell of 58.2 nm, and the low critical solution temperature is 42 °C, and the photothermal conversion efficiency increases from 14.04% to 58.41% by doping 1.6% of IR780 into the CFNPs-IR780 clusters. The dual-enzyme immobilized micro-systems and the carriers are recycled 12 and 72 times, respectively, and the enzyme activity remains above 70%. The micro-systems can realize whole recycling of the dual enzymes and carriers and further recycling of the carriers, thus providing a simple and convenient recycling method for dual-enzyme immobilized micro-systems. The findings reveal the micro-systems' important application potential in biological detection and industrial production.

The treatment of transarterial chemoembolization/hepatic arterial infusion chemotherapy combined with lenvatinib and PD-1 inhibitor is effective against hepatocellular carcinoma with portal vein tumor thrombus: A systematic review.

Background: Lenvatinib combined with programmed cell death protein-1 inhibitor has achieved good survival results in the treatment of hepatocellular carcinoma with portal vein tumor thrombus. Transarterial chemoembolization (TACE) or hepatic arterial infusion chemotherapy (HAIC) has attracted attention because of its high response rate and favorable survival rate in patients with liver cancer and portal vein tumor thrombus. This study aimed to compare the efficacy and safety of Lenvatinib combined with programmed cell death protein-1 inhibitor plus transarterial chemoembolization or hepatic arterial infusion chemotherapy in patients with hepatocellular carcinoma with portal vein tumor thrombus. Method: We searched PubMed, Embase and the Cochrane Library for studies. These included randomized controlled trials or clinical trials of Lenvatinib plus programmed cell death protein-1 inhibitor plus transarterial chemoembolization or hepatic arterial infusion chemotherapy (intervention group) versus Lenvatinib plus programmed cell death protein-1 inhibitor or Lenvatinib plus transarterial chemoembolization/hepatic arterial infusion chemotherapy or Lenvatinib alone (control group) in liver cancer with portal vein tumor thrombus The primary outcomes were overall survival and progression-free time, and the secondary outcomes were response rate and the rate of adverse events. According to the heterogeneity among different studies, Revman5.4 was used to conduct fixed effect or random effect model analysis. Results: Five clinical trials were included, including 311 cases in the intervention group and 309 cases in the control group. In terms of efficacy, compared with the control group, the overall survival (HR=1.88, 95%CI: 1.57-2.25, P < 0.00001) and progression-free survival (HR=1.62, 95%CI: 1.41-1.86, P < 0.00001), better efficacy, and better disease response than the control group. In terms of safety, the risk of treatment-related adverse events in the intervention group was higher than that in the control group, and White Blood cell count decreased (RR=0.72, 95%CI: 0.38-1.37, P=0.32), Platelet count decreased (RR=0.99, 95%CI: 0.65-1.51, P=0.96) and Total bilirubin increased (RR=0.86, 95%CI: Increased) 0.88-1.28, P=0.46) were lower than those in the control group, and the rest were higher than those in the control group, and the differences in some results were statistically significant. Conclusions: Transarterial chemoembolization or hepatic arterial infusion chemotherapy combined with Lenvatinib plus programmed cell death protein-1 inhibitor can effectively delay the progression, prolong the survival period and improve the quality of life of liver cancer patients with portal vein tumor thrombus.

ALK-JNK signaling promotes NLRP3 inflammasome activation and pyroptosis via NEK7 during Streptococcus pneumoniae infection.

Streptococcus pneumoniae (S. pneumoniae), a clinically important pathogen worldwide, causes serious invasive diseases, such as pneumonia, otitis media, and meningitis. The NLR family pyrin domain-containing 3 (NLRP3) inflammasome, an important component of the innate immune system, plays a key role in defense against pathogen infection; however the specific activation mechanism induced by S. pneumoniae infection is not fully understood. Here, primary mouse macrophages were selected as the in vitro cell model, and the effect of kinases on S. pneumoniae infection-induced NLRP3 inflammasome activation was investigated in vivo and in vitro using the western blot/RT-PCR/Co-IP/immunofluorescence staining/ELISA with or without kinase inhibitor or siRNA pretreatment. In this study, we found that the formation of the NEK7-NLRP3 complex significantly increased during S. pneumoniae infection and that anaplastic lymphoma kinase (ALK) and Jun N-terminal kinase (JNK) were phosphorylated rapidly. ALK and JNK inhibitors significantly reduced the ability of bacterial killing, the gene expression of NLRP3 inflammasome, the formation of apoptosis-associated speck-like protein containing caspase-recruitment domain (ASC) specks and the NEK7-NLRP3 complex, which in turn decreased the activation level of NLRP3 inflammasome-associated molecules and the maturation of interleukin-1ß (IL-1ß). In addition, ALK regulated the phosphorylation of JNK. Interestingly, the ALK/JNK/NEK7-NLRP3 signaling pathway is also involved in regulating pyroptosis and IL-1ß secretion triggered by S. pneumoniae infection. In conclusion, our data suggest, for the first time, that the ALK/JNK/NEK7-NLRP3 signaling pathway may play an important role in NLRP3 inflammasome activation and pyroptosis and consequently regulate the host immune response upon S. pneumoniae infection.

Identification of immuno-infiltrating MAP1A as a prognosis-related biomarker for bladder cancer and its ceRNA network construction.

Backgrounds: Approximately 75% of bladder cancer occurrences are of the non-muscle-invasive type. The estimated five-year survival rate is 26%-55%. Currently, there is no reliable biomarker available for early diagnosis and prognosis of bladder cancer. The present study aims to identify a biomarker using bioinformatic approaches to provide a new insight in clinical research for early diagnosis and prognosis of bladder cancer. Methods: Clinical data and a transcriptome of bladder cancer were obtained from TCGA, GEO, GETx, and UCSC Xena. The differential expressed gene (DEG) analysis, weighted gene co-expression network analysis (WGCNA), and survival analysis using the Kaplan-Meier and Cox proportional-hazards models were used to identify the Microtubule-associated Proteins 1A (MAP1A). on overall survival (OS) and disease-free survival (DFS) was analyzed using GEPIA and GETx databases. The TIMER 2.0 database predicted the correlation between MAP1A and immunocytes and immune checkpoints. Target prediction of the regulated competing endogenous RNAs (ceRNAs) network of MAP1A was performed using starBase and TargetScan. Cystoscope v3.7.2 software was used to visualize the ceRNA coexpression network. The R programming language v4.0.2 was applied as an analytic tool. Gene expression of MAP1A verified by RT-qPCR. Results: The low expression of MAP1A was verified in bladder cancer tissues and bladder cancer cell lines SW780 and 5637. P < 0.001 were obtained by Kaplan-Meier survival analysis and Cox proportional hazards model, with a hazard ratio (HR) of 1.4. Significant correlations between MAP1A and OS (P < 0.001, HR = 1.9) as well as DFS (P < 0.05, HR = 1.7) in bladder cancer were identified through gene expression profiling interactive analysis (GEPIA), indicating MAP1A may be a high-risk factor. Significant correlation in single copy-number variation of MAP1A gene with CD8+ T cells, and myeloid dendritic cells (MDCs) (P < 0.05) was noted. MAP1A expression was shown to be significantly correlated with the amount of CD4+ T cells and CD8+ T cells, MDCs, macrophages, and neutrophils in a statistically significant positive manner (P < 0.001). However, the MAP1A expression demonstrated a strong negative connection with B cells (P < 0.001). Except for macrophage M1 genes IRF5 and PTGS2, MAP1A expression was significantly correlated with the gene levels in immunocytes such as CD4+ T cells, CD8+ T cells, B cells, dendritic cells (DCs), macrophages, and neutrophils (Cor > 0.2, P < 0.001), as well as immune checkpoint related genes including cytotoxic t-lymphocyte-associated protein 4 (CTLA-4), programmed death 1 (PD-1), programmed death ligand 1 (PD-L1) (P < 0.001). Finally, we predicted that the MAP1A-interacting miRNA was miR-34a-5p, and the MAP1A endogenous competing RNAs were LNC00667, circ_MAP1B, and circ_MYLK, respectively. These findings support the need for further studies on the mechanism underlying the pathogenesis of this disease. Conclusion: MAP1A is considered as a prospective biomarker for early diagnosis, therapeutic observation, and prognosis analysis in bladder cancer.

Short-term effectiveness and safety of CT-guided radioactive iodine-125 seed implantation for treatment of adrenal metastases.

Purpose: To evaluate short-term effectiveness and safety of computed tomography (CT)-guided radioactive iodine-125 (125I) seed implantation (CTRISI) for treating adrenal metastases. Material and methods: A total of 50 consecutive patients with adrenal metastases were enrolled retrospectively. Among them, 18 patients received CTRISI, and 18 received 3D-conformal radiotherapy (3D-CRT) treatment. The remaining 14 patients without any treatments served as a control group. Follow-up CT was performed at 6 weeks, 3 months, and 6 months after treatment. Tumor responses and complications were evaluated. Results: At 6 weeks, control rate in control group (complete response [CR] + partial response [PR]) was 0, and in the CTRISI group (CR + PR, 84.41%), it was significantly higher than that in the 3D-CRT group (CR + PR, 44.44%). Local control rates with CTRISI at 3 and 6 months were 68.42% and 57.89%, respectively. No severe complications were observed after CTRISI. Conclusions: CTRISI is an effective and safe method for short-term treatment of adrenal metastases. Our findings suggest that CTRISI can safely and effectively be used for adrenal metastases patients as short-term treatment. Further survival studies with longer follow-up are warranted to validate our results.

Doxorubicin Loaded Dextran-coated Superparamagnetic Iron Oxide Na-noparticles with Sustained Release Property: Intracellular Uptake, Phar-macokinetics and Biodistribution Study.

BACKGROUND: Due to the short biological half-life and serious side effects (especially for heart and kidney), the application of Doxorubicin (Dox) in clinical therapy is strictly limited. To overcome these shortcomings, a novel sustained release formulation of doxorubicin-loaded dextran-coated superparamagnetic iron oxide nanoparticles (Dox-DSPIONs) was prepared. OBJECTIVE: The purpose of this study was to evaluate the intracellular uptake behavior of Dox-DSPIONs and to investigate their pharmacokinetics and biodistribution properties. METHOD: Confocal laser scanning microscopy was employed to study the intracellular uptake and release properties of Dox from Dox-DSPIONs in SMMC-7721 cells. Simple high-performance liquid chromatography with fluorescence detection (HPLC-FLD) method was established to study the pharmacokinetics and biodistribution properties of Dox-DSPIONs in vivo after intravenous administration and compared with free Dox. RESULTS: Intracellular uptake experiment indicated that Dox could be released sustainedly from Dox-DSPIONs over time. The pharmacokinetics parameters displayed that the T1/2and AUC0-24h of Dox-DSPIONs were higher than those of free Dox, while the Cmax of Dox-DSPIONs was significantly lower than that of free drug. The biodistribution behaviors of the drug were altered by Dox-DSPIONs in mice, which showed obvious liver targeting, and significantly reduced the distribution of the drug in the heart and kidney. CONCLUSION: Dox-DSPIONs have the sustained-release property in vitro and in vivo, which could significantly prolong blood circulation time, improve bioavailability, and reduce the side effects of Dox. Therefore, the novel formulation of the Dox-DSPIONs has the potential as a promising drug delivery system in cancer therapy.

The role of morphology, shell composition and protein corona formation in Au/Fe3O4 composite nanoparticle mediated macrophage responses.

The great interest in using nanoparticles (NPs) for biomedical applications is transversal to various materials despite the poorly understood correlation between their physicochemical properties and effects on the immune system. NPs, such as gold and Fe3O4, are generally regarded as safe, but the immunotoxicological profile of Au/Fe3O4 composite NPs with different physicochemical properties is not well documented. This study investigated the biological impact of Au/Fe3O4 composite NPs with different morphologies (spherical core-shell and flower-like) and shell composition in vitro to analyze their potential cytotoxic effects and inflammatory responses on RAW 264.7 cells. Au/Fe3O4 composite NPs with a flower-like structure (FLNPs) induce a pronounced reduction in cell viability compared with Au/Fe3O4 composite NPs with a spherical core-shell structure (CSNPs). The increased production of reactive oxygen species, which damages cellular membranes, might contribute to the cytotoxicity effect of FLNPs. However, CSNPs presented more RAW 264.7 cell adhesion and uptake than FLNPs. Remarkably, a significant TNF-α release was observed with CSNP treated RAW 264.7 cells other than that of FLNPs. Protein corona analysis revealed the adsorption of a distinct amount and profile of proteins on the surface of CSNPs and FLNPs. Given the similar particle size and ζ-potential of CSNPs and FLNPs under the cell culture condition, results indicate that the impact of Au/Fe3O4 composite NPs on the macrophage activity highly depends on their morphology, shell composition and protein corona profile.

Lateral flow immunoassay based on gold magnetic nanoparticles for the protein quantitative detection: Prostate-specific antigen.

Point-of-care testing (POCT) demands for rapidly obtaining test results by means of portable analytical instruments and auxiliary reagents at the sampling site. It's important for tumor marker to be recognized and detected in early clinical diagnosis. Many studies focused on producing small portable devices that would allow fast, accurate, and on-site detection. This study aimed to report a magnetic quantitative lateral flow immunoassay (LFIA) system based on poly (acrylic acid) (PAA)-modified gold magnetic nanoparticles (PGMNs) for detecting prostate-specific antigen (PSA) qualitatively and quantitatively. The result was easily achievable with a portable magnetic reader within 15 min. Under optimal conditions, as low as 0.17 ng/mL PSA could be detected. The method was validated using a well-established Solin electrochemiluminescence immunoassay and showed high consistency in detecting 84 serum samples (R2 = 0.98). The quantitative LFIA based on PGMNs established in this study was proven to be rapid, accurate, sensitive, and inexpensive. As a POCT, it can be potentially developed for the quantitative diagnosis of other disease-related protein biomarkers.

The marker of cobalamin deficiency, plasma methylmalonic acid, may help identifying lysosomal iron trapping in patients. Its possible utility for heart failure.

Iron deficiency is known to aggravate the prognosis of patients with heart failure. Iron has functions in the mitochondrial respiratory chain. In patients with reduced mitochondrial respiration, the mitochondrial ratio between the level of nicotinamide adenine dinucleotide and its reduced form decreases. Due to the mitochondrial-lysosomal interplay, decreased mitochondrial respiration also leads to inhibition of lysosomal hydrolysis. As a result, cobalamin and iron will be trapped in lysosomes. This will, even if iron and cobalamin have been consumed and absorbed in sufficient amounts, lead to their functional deficiencies.1 Functional iron deficiency can further impede mitochondrial respiration. Increased plasma levels of methylmalonic acid were shown to predict all-cause and cardiovascular mortality in the general population. Treatments targeting mitochondrial and lysosomal function may correct the functional deficiencies and improve prognosis in a subgroup of patients with heart failure, notably those with skeletal muscle wasting. Methylmalonic acid levels may be used for monitoring response to treatment, thereby identifying patients of the subgroup in which disease outcome may improve.

A site-specific DNA methylation biosensor for both visual and magnetic determination based on lateral flow assay.

Tumorigenesis driven by abnormal DNA methylation has highlighted the need to develop a portable, rapid and sensitive strategy for accurate methylation detection with a specific cancer-prognostic gene, which caters to the popularization of precision medicine. In this study, a site-specific biosensor for both visual and magnetic DNA methylation determination has been established based on lateral flow assay. By introducing digoxin- and biotin-labeled primers into PCR, the amplicons can be recognized and captured by gold magnetic nanoparticles (GMNPs) in this biosensor. Working as a signal probe, the optical property of GMNPs allows the amplicons to be interpreted with naked eyes avoiding any complex equipment and cumbersome operation after PCR. Moreover, by virtue of the magnetic property of GMNP, the signal can be explained and recorded by a magnetometer in clinical practice. The introduction of tailor-made primer sets makes it possible to accurately distinguish 0.1% methylated variants in the presence of numerous unmethylated variants as strong interferential background and vice versa at target cytosine-guanine dinucleotide. A distinct signal can be observed with as low as 0.01 pg variants for both visual and magnetic analyses. As a significant tumor suppressor gene, the promoter methylation status of miR-34a is accurately determined with not only cell lines but also with clinical samples, which demonstrates the great potential of this biosensor for cancer diagnosis and prognosis.

One-step determination of deletion mutation based on loop-mediated isothermal amplification.

Deletion mutation has been proved as the important factor for occurrence and development of disease, especially those with cancer. With the popularity of precision medicine, the individual cancer therapeutic strategy has highlighted the requirement to develop a straightforward and competent strategy for deletion mutation determination. Hence, the present study is dedicated to develop a one-step assay to identify deletion mutation with sequence specificity for clinical practice. Taking advantage of loop-mediated isothermal amplification, an ultrasensitive and rapid deletion mutation determination method is established, which allow as low as 30 copies or 0.1% target variants under strong interferential background can be accurately distinguished in 30 min dispensing with professional operation and complex data interpretation. As a demonstration, the epidermal growth factor receptor p.E746-A750del, a crucial factor for the susceptibility of tyrosine kinase inhibitor in non-small-cell lung cancer treatment, has been accurately identified by this method with both cell lines and real clinical samples. By tailor-made primer set, this method can be extended for other deletion mutants, making it a molecular diagnostic tool and could be readily adapted for cancer diagnosis, therapy and prognosis in point of care diagnostic test scenario.

A fluorescein-coumarin based ratiometric fluorescent probe for detecting hydrazine and its real applications in cells imaging.

According to the mechanism of hydrazine to ester bond elimination, a novel ratiometric fluorescent probe (FCP) based on the fluorescein-coumarin structure is designed and synthesized for detecting hydrazine. The obvious red shift in the absorption and fluorescence spectrum is caused by the hydrolysis of the ester bond of FCP by upon mixing with hydrazine. The proposed FCP probe is shown to have linear detection ranges from 0 to 250 nM for hydrazine, and LOD is 0.364 nM. In addition, this promising probe possesses high sensitivity and selectivity for monitoring the intracellular hydrazine, thus, has great potential to be applied in early diagnosis of disease. Meanwhile, it has been successfully applied to the detection and cell imaging of hydrazine in actual water samples.

Functional Compensation and Mechanism of Choline Acetyltransferase in the Treatment of Cognitive Deficits in Aged Dementia Mice.

Choline acetyltransferase (ChAT) synthesizes the neurotransmitter acetylcholine (Ach). Exogenous supplementation with ChAT can functionally compensate for decreased Ach levels and ameliorate memory and cognitive deficits. In this paper, the treatment efficacy of recombinant ChAT (peptide transduction domain (PTD)-ChAT) and donepezil were compared in aged dementia mice, and their mechanisms were explored by performing the gene function annotation and enrichment analysis of differentially expressed genes. The Morris water maze test showed that the swimming times of PTD-ChAT-treated (4 mg/kg) and donepezil-treated (0.5 mg/kg) mice with mild and moderate dementia were significantly shortened (P < 0.01 vs aged dementia mice), and no significant changes were observed between the PTD-ChAT- and donepezil-treated groups. In contrast, the swimming times of PTD-ChAT-treated mice with severe dementia were noticeably shorter than those of donepezil-treated mice with severe dementia (P < 0.01), indicating that the treatment efficacy of PTD-ChAT is superior to that of donepezil. The effect of PTD-ChAT was further confirmed in transgenic dementia mice (C57BL/6J-TgN (APP/PS1) ZLFILAS). Gene function annotation and enrichment analysis showed that PTD-ChAT improved cognitive deficits through Ach and was implicated in neuroprotection, synaptic plasticity, neuronal survival, and cerebrovascular remodeling through ACh and vascular endothelial growth factor (VEGF) pathway activation. Donepezil was significantly correlated with the immune inflammatory response and the insulin and IGF-1 signaling pathways. Therefore, although PTD-ChAT and donepezil were both effective in the treatment of aged dementia mice, their mechanisms were significantly different. Our research indicated that PTD-ChAT has potential promise for research on new drugs for AD treatment.

Long non­coding RNA CASC2 enhances berberine­induced cytotoxicity in colorectal cancer cells by silencing BCL2.

Berberine, a natural isoquinoline alkaloid derived from Berberis species, has been reported to have anticancer effects. However, the mechanisms of action in human colorectal cancer (CRC) are not well established to date. In the present study, the cell cytotoxicity effect of berberine on human CRC cells, as well as the possible mechanisms involved, was investigated. The results of the cell viability and apoptosis assay revealed that treatment of CRC cells with berberine resulted in inhibition of cell viability and activation of cell apoptosis in a concentration­dependent manner. To reveal the underlying mechanism of berberine­induced anti­tumor activity and cell apoptosis, RNA­sequencing followed by reverse­transcription quantitative PCR were performed. In addition, RNA immunoprecipitation, chromatin immunoprecipitation and western blot analysis were used to identify the functional regulation of CASC2/EZH2/BCL2 axis in berberine­induced CRC cell apoptosis. The data revealed that lncRNA CASC2 was upregulated by berberine treatment. Gain­ or loss­of­function assays suggested that lncRNA CASC2 was required for the berberine­induced inhibition of cell viability and activation of cell apoptosis. Subsequently, the downstream antiapoptotic gene BCL2 was identified as a functional target of the berberine/CASC2 mechanism, as BCL2 reversed the berberine/CASC2­induced cell cytotoxicity. lncRNA CASC2 silenced BCL2 expression by binding to the promoter region of BCL2 in an EZH2­dependent manner. In summary, berberine may be a novel therapeutic agent for CRC and lncRNA CASC2 may serve as an important therapeutic target to improve the anticancer effect of berberine.

Quercetin-Modified Metal-Organic Frameworks for Dual Sensitization of Radiotherapy in Tumor Tissues by Inhibiting the Carbonic Anhydrase IX.

The development of multifunctional nanoscale radiosensitizers has attracted a tremendous amount of attention, which can enhance the radiosensitization of tumor tissues and reduce unnecessary damage to the surrounding organs. However, the persistent hypoxia environment within the tumor limits their applications in radiotherapy. In this paper, a stable nanocomposite was engineered to overcome the hypoxia properties by using 1,4-benzenedicarboxylic acid produced from a Zr-MOF as a carbonic anhydrase IX (CA IX) inhibitor and quercetin (QU) as a radiosensitizer. QU was encapsulated into the Zr-MOF structure to achieve a synergetic dual sensitization therapy. Zr-MOF-QU exhibits an excellent potential of radiotherapy sensitization characteristics in vitro and in vivo from the γ-H2AX immunofluorescence staining and colony assays. The mechanisms of alleviating hypoxia-induced resistance and sensitizing tumor tissues to improve cell apoptosis from radiation were found to suppress CA IX expressions by the decomposition product from Zr-MOF and boost the sensitivity by QU in radiation therapy. Moreover, there was no significant systemic toxicity during the treatment, and the therapeutic outcome was assessed in animal models. Therefore, our results demonstrate a promising cancer treatment approach in the radiation field.

Berberine Promotes Apoptosis of Colorectal Cancer via Regulation of the Long Non-Coding RNA (lncRNA) Cancer Susceptibility Candidate 2 (CASC2)/AU-Binding Factor 1 (AUF1)/B-Cell CLL/Lymphoma 2 (Bcl-2) Axis.

BACKGROUND Berberine, a natural isoquinoline alkaloid derived from Berberis genus plants, has been reported to have anti-cancer effects. While cell behavior can be modulated by long non-coding RNAs (lncRNAs), the contributions of lncRNAs in progression and berberine effects on colorectal cancer are largely unknown. Therefore, the present study investigated the involvement and regulatory function of lncRNA cancer susceptibility candidate 2 (CASC2) during the treatment of human colorectal cancer using berberine. MATERIAL AND METHODS Reverse transcription-quantitative PCR (RT-qPCR) was performed to detect the expression levels of lncRNA CASC2 and Bcl-2 mRNA in colorectal cancer cells. MTT assay was performed to evaluate cell viability. Flow cytometry and TUNEL assay were used to analyze the apoptosis of cancer cells. RNA immunoprecipitation (RIP) assay was done to verify the interaction between lncRNA CASC2 and (AU-binding factor 1) AUF1, or AUF1 and B-cell CLL/lymphoma 2 (Bcl-2). RESULTS Treatment with berberine suppressed cell viability of colorectal cancer by promoting apoptosis level. LncRNA CASC2 was upregulated in cells treated with berberine, and knockdown of lncRNA CASC2 reversed the berberine-induced apoptosis. In addition, anti-apoptotic gene Bcl-2 was suppressed by berberine treatment and lncRNA CASC2, inducing the pro-apoptotic effects. Moreover, lncRNA CASC2 binds to AUF1, which sequestered AUF1 from binding to Bcl-2 mRNA, thus inducing the inactivation of Bcl-2 translation. CONCLUSIONS Our study reveals that lncRNA CASC2 mediates the berberine-induced pro-apoptotic effect via inhibition of Bcl-2 expression at the post-transcriptional level.