Suv39h1 contributes to activation of hepatic stellate cells in non-alcoholic fatty liver disease by enabling anaerobic glycolysis.

AIMS: Non-alcoholic fatty liver disease (NAFLD) has become a global epidemic. Excessive fibrogenesis, characterized by activation of hepatic stellate cells (HSCs), is a hallmark event in late stages of NAFLD. HSC activation is metabolically programmed by anaerobic glycolysis. In the present study we investigated the involvement of suppressor of variegation 3-9 homolog 1 (Suv39h1), a lysine methyltransferase, in NAFLD-associated liver fibrosis. METHODS AND MATERIALS: Liver fibrosis was induced by feeding the mice with a methionine-and-choline deficient (MCD) diet for 8 weeks. RESULTS: We report that germline deletion of Suv39h1 attenuated liver fibrosis in mice fed an MCD diet. In addition, HSC conditional deletion of Suv39h1 similarly ameliorated liver fibrosis in the NAFLD mice. Interestingly, co-culturing with hepatocytes exposed to palmitate promoted glycolysis in wild type HSCs but not in Suv39h1 deficient HSCs. Mechanistically, Suv39h1 facilitated the recruitment of hypoxia induced factor (HIF-1α) to stimulate the transcription of hexokinase 2 (HK2) in HSCs thereby enhancing glycolysis. Importantly, a positive correlation between Suv39h1, HK2, and myofibroblast markers was identified in liver specimens from NAFLD patients. SIGNIFICANCE: In conclusion, our data identify a novel pathway that contributes to the liver fibrosis and points to the possibility of targeting Suv39h1 for the intervention of liver fibrosis in NAFLD.

Investigations on ultrasonography in the diagnosis of nodular localized cutaneous neurofibroma.

OBJECTIVE: To describe the ultrasound characteristics of nodular localized cutaneous neurofibroma (NLCN). MATERIALS AND METHODS: Clinical features and ultrasound characteristics of 43 lesions of 40 patients pathologically proven as NLCNs at Peking University Shenzhen Hospital from October 2014 to May 2022 were analyzed retrospectively. The location, length-to-thickness (L/T) ratio, thickness-to-width (T/W) ratio, shape, margin, capsule, echogenicity, echotexture, posterior features, vascularity, and "rat tail sign" were evaluated. RESULTS: All ultrasound findings showed almost perfect agreement. More than a half of NLCNs (n = 24, 55.8%, p < 0.001) were located in the subcutaneous fat layer wholly with well-demarcation from dermis and deep fascia. Most of the NLCNs were fusiform shape (n = 27, 62.8%, p < 0.001) in the long axis and oval shape (n = 35, 81.4%, p < 0.001) in the short axis. The other ultrasound findings of NLCNs included well-defined (n = 42, 97.7%, p < 0.001), encapsulated (n = 39, 90.7%, p < 0.001), predominately hypoechoic (n = 34, 79.1%, p < 0.001), homogeneous (n = 39, 90.7%, p < 0.001), posterior enhancement (n = 29, 67.4%, p = 0.033), and avascularity (n = 37, 86.0%, p < 0.001). Only a quarter (n = 11, 25.6%, p = 0.002) of lesions were recognized with the "rat tail sign." CONCLUSION: NLCNs present as fusiform shape in long axis and round shape in short axis. The common ultrasound findings of NLCNs are well-defined, encapsulated, predominately hypoechoic, homogeneous lesion with posterior enhancement, and poor blood supply. The "rat tail sign" has low sensitivity in NLCNs.

Mechanisms of Panax Ginseng on Treating Sepsis by RNA-Seq Technology.

Purpose: To explore the potential active targets and mechanisms of Panax Ginseng in the treatment of sepsis using network pharmacology and RNA-seq technology. Patients and Methods: Patients with sepsis and healthy volunteers were collected according to SEPSIS 3.0, and their peripheral blood was used for RNA-seq analysis. The active ingredients and targets of Panax Ginseng were obtained using the TCMSP database, PPI and GO analysis were performed for disease-drug intersection targets. Then, we used Meta-analysis to screen core genes. Finally, single-cell RNA-seq was used to perform cell localization analysis on core genes. Results: RNA-seq analysis collected 4521 sepsis-related genes, TCMSP database obtained 86 Panax Ginseng active ingredients and their 294 active targets. PPI and GO analysis showed intersection targets were closely linked, and mainly involved in cellular response to chemical stress, response to drug and molecule of bacterial origin, etc. Then, core targets, IL1B, ALOX5, BCL2 and IL4R, were sorted by Meta-analysis, and all four genes have high expression in the sepsis survivor group compared to the sepsis non-survivor group; single-cell RNA-seq revealed that IL1B was mainly localized in macrophages, ALOX5 was mainly localized in macrophages and B cells, BCL2 was mainly localized in natural killer cells, T cells and B cells, IL4R was widely distributed in immune cells. Finally, according to the correspondence between the active ingredients and targets of Panax Ginseng in TCMSP database, we found that Ginsenoside rh2 regulates the expression of IL1B, Ginsenoside rf regulates the expression of IL1B and IL4R, Kaempferol regulates the expression of ALOX5 and BCL2, and ß-sitosterol regulates the expression of BCL2. Conclusion: Ginsenoside rh2, Ginsenoside rf, Kaempferol and ß-sitosterol may produce anti-sepsis effects by regulating the expression of IL1B, ALOX5, BCL2 and IL4R, thus improving the survival rate of sepsis patients.

The Role of Praziquantel in the Prevention and Treatment of Fibrosis Associated with Schistosomiasis: A Review.

Schistosomiasis remains a major global public health concern. Currently, the control of this neglected tropical disease still depends on chemotherapy to reduce the prevalence and intensity of the parasite infection. It has been widely accepted that praziquantel is highly effective against all species of Schistosoma, and this agent is virtually the only drug of choice for the treatment of human schistosomiasis. Mass drug administration (MDA) with praziquantel has been shown to be effective in greatly reducing the prevalence and morbidity due to schistosomiasis worldwide. In addition to antischistosomal activity, a large number of experiential and clinical evidence has demonstrated the action of praziquantel against fibrosis caused by S. mansoni and S. japonicum infections through decreasing the expression of fibrotic biomarkers such as α-smooth muscle actin (α-SMA), collagen, matrix metalloproteinase (MMP), and tissue inhibitor of metalloproteinase (TIMP), and inhibiting the expression of proinflammatory cytokines such as interleukin (IL)-6, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-ß, as well as chemokines, and similar antifibrotic activity was observed in mouse models of fibrosis induced by carbon tetrachloride (CCl4) and concanavalin A (Con-A). In this review, we discuss the role of praziquantel in the prevention and treatment of fibrosis associated with schistosomiasis and the possible mechanisms. We call for randomized, controlled clinical trials to evaluate the efficacy and safety of praziquantel in the treatment of schistosomiasis-induced hepatic fibrosis, and further studies to investigate the potential of praziquantel against fibrosis associated with alcohol consumption, viruses, and toxins seem justified.

Oridonin-loaded lipid-coated calcium phosphate nanoparticles: preparation, characterization, and application in A549 lung cancer.

In this study, the feasibility of using the novel anisamide-lipid calcium phosphate nanoparticles (AS-LCPs) as a nanocarrier for the delivery of biologically active oridonin (ORD) was evaluated on lung cancer models. In addition to the characterization, release behaviors, and stability of AS-ORD LCPs, we also studied their pharmacokinetics and targeting ability by in vivo imaging. Finally, we investigated the effect of ORD on the anti-tumor efficiency of the AS-LCPs based on weight, tumor inhibition, and the survival time of mice. The average particle size of the AS-ORD LCPs was 129.5 ± 23.7 nm, the polydispersity index (PDI) was 0.16 ± 0.03, and the zeta potential was 23.6 ± 3.4 mV. The AS-ORD LCPs were proved to be stable under both long-term and accelerated storage conditions. The AS-ORD LCPs showed sustained release in vivo and faster release in acidic environment, which was favorable to drug release in tumor environment. In vivo studies showed that depending on surface modification, AS-ORD LCPs which suggested that cell internalization was change and more drugs entered the cells successfully. Therefore, AS-ORD LCPs could be a promising formulation for the treatment of lung cancer.

TRIM46 activates AKT/HK2 signaling by modifying PHLPP2 ubiquitylation to promote glycolysis and chemoresistance of lung cancer cells.

The incidence of lung cancer is increasing worldwide. Although great progress in lung cancer treatment has been made, the clinical outcome is still unsatisfactory. Tripartite motif (TRIM)-containing proteins has been shown to be closely related to tumor progression. However, the function of TRIM46 in lung cancer is largely unknown. Here, TRIM46 amplification was found in lung adenocarcinoma (LUAD) tissues and TRIM46 amplification was significantly associated with a poor survival rate. Overexpression of wild type TRIM46 increased the proliferation of LUAD cells and glycolysis, promoted xenografts growth, and enhanced cisplatin (DDP) resistance of LUAD cells via increased ubiquitination of pleckstrin homology domain leucine-rich repeat protein phosphatase 2 (PHLPP2) and upregulation of p-AKT. In contrast, overexpression of RING-mutant TRIM46 did not show any effects, suggesting the function of TRIM46 was dependent on the E3 ligase activity. Furthermore, we found that TRIM46 promoted LUAD cell proliferation and DDP resistance by enhancing glycolysis. PHLPP2 overexpression reversed the effects of TRIM46 overexpression. Amplification of TRIM46 also promoted LUAD growth and enhanced its DDP resistance in a patient-derived xenograft (PDX) model. In conclusion, our data highlight the importance of TRIM46/PHLPP2/AKT signaling in lung cancer and provide new insights into therapeutic strategies for lung cancer.

Co-Delivery Anticancer Drug Nanoparticles for Synergistic Therapy Against Lung Cancer Cells.

INTRODUCTION: This study aims to develop a novel co-delivery gefitinib and quercetin system loaded with PLGA-PEG nanoparticles and evaluate their antitumor activity in vitro and in vivo. METHODS: Gef/Qur NPs were prepared and characterized. The release of drugs, stability, cellular uptake and cytotoxicity were evaluated in vitro. The antitumor effects and systemic toxicity of different formulations were also investigated. RESULTS: Gef/Qur NPs displayed a smaller particle size and a PDI and zeta potential of 0.11 and -23.5 mV, respectively. The hydrophobic Gef and Qur content in NPs reached up to 65.2% and 56.4%, respectively, and their high entrapment efficiencies recorded 83.7% and 82.3%, respectively. The in vitro release of Gef/Qur from the NPs was sustained for 12 h. Compared with control groups, Gef/Qur NPs showed higher cellular uptake and cell inhibition rates. In vivo studies identified the lungs as the target tissue and the region of maximum drug release. Through pharmacodynamics analysis, we found that two drugs (Gef and Qur) were incorporated into one nanoparticle carrier, which played a good role in generating synergistic effect. DISCUSSION: It is concluded that PLGA-PEG is an ideal drug carrier for the co-delivery of Gef/Qur to treat lung cancer.

Gene regulation and prognostic indicators of lung squamous cell carcinoma: TCGA-derived miRNA/mRNA sequencing and DNA methylation data.

Lung squamous cell carcinoma (LSCC) is a common cancer worldwide, and this study aimed to investigate the key regulatory networks and prognostic indicators of LSCC. MicroRNA (miRNA)/messenger RNA (mRNA) sequencing and DNA methylation data were obtained from the Cancer Genome Atlas. Differentially expressed miRNAs (DEmiRNAs) and genes (DEGs) were identified by the limma package. Then, the transcription factors (TFs) of DEmiRNAs/DEGs, as well as the targets of miRNAs, were predicted by the TFmiR online tool. Using the t test, aberrant methylation was detected in TF binding sites (TFBSs) in promoters. Finally, integrated network and survival analyses were conducted using SPSS software. We obtained 104 DEmiRNAs and 4,491 DEGs, and validated 2,113 DEGs (VDEGs). Then, 103 TFs, 295 TFs, and 14 DEmiRNAs were predicted to target 95 DEmiRNAs, 821 DEGs and 283 DEGs, respectively. After TF-DEmiRNA/DEG and TF-DEmiRNA-DEG networks were constructed (e.g., E2F1-CDC25A, miR29a-RAN, miR326-TBL1XR1), five feedforward loops between ZEB1 and miR-141/200a/200b/200c/429 were found. Furthermore, VDEGs CDC25A, RAN, TBL1XR1 as well as miR-130b and miR-590 were negatively correlated with survival rates. E2F1-CDC25A, miR29a-RAN, miR326-TBL1XR1, and the feedforward loops between ZEB1/ZEB2 and miR-141/200a/200b/200c/429 might participate in LSCC development. Compared with BEAS-2B cells, the SK-MES-1 cells presented a higher expression level of miR-141, miR-200a, miR-200b, miR-200c but a lower expression level of ZEB1. Overexpressed miR-200c significantly attenuated the expression of ZEB1 and ZEB2 and inhibited the proliferation and migration of SK-MES-1 cells (all p < 0.05). In addition, CDC25A, miR-200a, miR-200b, miR-200c, miR-130b, and miR-590 are potential prognostic indicators of LSCC.

Knockdown of TRIM65 inhibits autophagy and cisplatin resistance in A549/DDP cells by regulating miR-138-5p/ATG7.

Cisplatin resistance is the main cause of treatment failure in patients with non-small-cell lung cancer (NSCLC). Autophagy is a key mechanism of resistance to chemotherapy. Given that tripartite motif (TRIM)-containing proteins are involved in the regulation of autophagy and chemoresistance, we aimed to study the functions of TRIM protein members in autophagy-mediated chemoresistance of NSCLC. We found that TRIM65 was significantly increased in cisplatin-resistant NSCLC cell line (A549/DDP) as compared to the parental cell line (A549). Knockdown of TRIM65 can enhance cisplatin-induced apoptosis and inhibit autophagy in A549/DDP cells, as indicated by Annexin V/PI staining, caspase3 activity test, and LC3-II immunofluorescence staining. Additionally, knockdown of TRIM65 significantly decreased the expression of an important autophagy mediator, ATG7, which was a potential target of miR-138-5p. miR-138-5p inhibitor significantly abolished the effects of TRIM65 knockdown on autophagy and cisplatin-induced apoptosis. Moreover, TRIM65 induced the ubiquitination and degradation of TNRC6A, resulting in the suppressed expression of miR-138-5p. TRIM65 knockdown inhibited the growth of tumors derived from A549/DDP cells. Furthermore, cisplatin-resistant NSCLC tissues displayed higher expression of TRIM65 mRNA and lower expression of miR-138-5p as compared to cisplatin non-resistant ones. miR-138-5p expression was negatively correlated with TRIM65 mRNA in NSCLC tissues. Collectively, the present study indicates that TRIM65 knockdown attenuates autophagy and cisplatin resistance in A549/DDP cells via regulating miR-138-5p.

HA/HSA co-modified erlotinib-albumin nanoparticles for lung cancer treatment.

BACKGROUND: Aim of this study was to prepare the hyaluronic acid and human serum albumin modified erlotinib nanoparticles (ERT-HSA-HA NPs) delivery system by a precipitation method. METHODS: ERT-HSA-HA NPs were characterized for physical properties, such as morphology and particle size, and in vitro drug release. Moreover, the cytotoxicity, cellular uptake, in vivo studies of ERT-HSA-HA nanoparticle were investigated and compared in A549 cells. RESULTS: The ERT-HSA-HA NPs showed spherical morphology, and their hydrodynamic diameter was 112.5±2.8 nm. The drug loading amount and encapsulation efficiency were 5.6% and 81.2%, respectively. After 3 months of storage, no dramatic change, such as visible aggregation, drug content changes, and precipitation, in the appearance of ERT-HSA-HA NPs occurred. In vitro release showed that the release of ERT from HSA-HA NPs was slow, without obvious burst effects at an early stage. In in vivo studies, ERT-HSA-HA NPs showed a superior antiproliferative effect on A549 cells, and the HA modification strategy can also facilitate the high-efficiency uptake of ERT-HSA NPs by A549 cells. Pharmacokinetic studies showed that the form of NPs could significantly extend the role of ERT in vivo (provided higher bioavailability). However, there was no significant difference in the pharmacokinetic parameters between ERT-HSA NPs and ERT-HSA-HA NPs after intravenous administration. In terms of in vivo antitumor activity, ERT-HSA-HA NP-treated mice showed a significantly suppressed tumor growth and no relapse after 30 d of treatment. CONCLUSION: HA/HSA co-modified erlotinib albumin nanoparticles was expected to be a new strategy in the treatment of lung cancer.

Vortex-assisted liquid-liquid microextraction of strontium from water samples using 4',4″(5″)-di-(tert-butylcyclohexano)-18-crown-6 and tetraphenylborate.

A vortex-assisted liquid-liquid microextraction method was developed for the chromatographic determination of strontium in aqueous samples. In the method, strontium was complexed with 4',4″(5″)-di-(tert-butylcyclohexano)-18-crown-6 in the presence of tetraphenylborate as the counter anion, which increased the hydrophobicity of the ion-association complex, resulting in its improved extraction into 1-octanol. Strontium from the organic phase was stripped with nitric acid back to aqueous solution and determined by ion chromatography. The optimum microextraction conditions were as follows: 2.0 mL aqueous samples with 3 mM tetraphenylborate; 150 µL of 1-octanol as the extractant phase with 10 mM DtBuCH18C6; vortex extraction time for 10 s; centrifugation at 6000 rpm for 4 min; stripping by 0.1 M nitric acid. Under the optimum conditions, the detection limit for strontium was 0.005 mg/L. The calibration curves showed good linearity over the range between 0.01 and 2.5 mg/L. Intra- and interday precisions of the present method were satisfactory with relative standard deviations of 1.7 and 2.1%, respectively.

Clinicopathological features and prognosis of primary pulmonary lymphoepithelioma-like carcinoma.

BACKGROUND: Lymphoepithelioma-like carcinoma (LELC) is a rare form of non-small cell lung carcinoma. The current study focused on its clinicopathological features and potential factors influencing the prognosis. METHODS: The statistical analysis was based on the clinicopathological records and the prognosis of 43 LELC patients, analyzed by Kaplan-Meier method, Log-rank test, and COX regression analysis. RESULTS: The patients' average age was 57.35±9.22 years, 86.05% of them were non-smokers and 53.49% were women. The average tumor diameter was 3.24±1.57 cm. The 2- and 5-year overall survival (OS) rates of LELC patients were 90% and 74%, respectively; the disease-free survival (DFS) rates were 87% and 47%, respectively. The patients with large tumor, accompanied with lymph nodes metastasis or at the advanced stage had the worst OS, and the patients with lymph nodes metastasis or at the advanced stage had the worst DFS. Univariate analysis indicated that T and N grading and TNM stage influenced the OS, and N grading and TNM stage influenced the DFS; the independent factors affecting OS or DFS were not identified by multivariate analysis. CONCLUSIONS: LELC commonly occurred in senior non-smoking women. In summary, the prognosis of LELC was satisfactory.

Ranking candidate genes of esophageal squamous cell carcinomas based on differentially expressed genes and the topological properties of the co-expression network.

BACKGROUND: The aim of this study was to identify the candidate genes of esophageal squamous cell carcinoma (ESCC). METHODS: Gene expression profiling of 17 ESCC samples and 17 adjacent normal samples, GSE20347, was downloaded from Gene Expression Omnibus database. The raw data were preprocessed, and the differentially expressed genes (DEGs) between ESCC and normal samples were identified by using SAM software (false discovery rate <0.001). Then, the co-expression network of DEGs was constructed based on Pearson's correlation test (r-value ≥0.8). Furthermore, the topological properties of the co-expression network were analyzed through NetworkAnalyzer (default settings) of Cytoscape. The expression fold changes of DEGs and topological properties were utilized to identify the candidate genes of ESCC (Crin score >4), which were further analyzed based on DAVID functional enrichment analysis (P-value <0.05). RESULTS: A total of 1,063 DEGs were identified, including 490 up-regulated and 573 down-regulated DEGs. Then, the co-expression network of DEGs was constructed, containing 999 nodes and 46,323 edges. Based on the expression fold changes of DEGs and the topological properties of the co-expression network, DEGs were ranked, and the top 24 genes were candidate genes of ESCC, such as CRISP3, EREG, CXCR2, and CRNN. Furthermore, the 24 genes were significantly enriched in bio-functions regarding cell differentiation, glucan biosynthetic process and immune response. CONCLUSION: The present study suggested that CRISP3, EREG, CXCR2, and CRNN might be causative genes of ESCC, and play vital roles in the development of ESCC. However, further experimental studies are needed to confirm our results.

The histone demethylase PHF8 is an oncogenic protein in human non-small cell lung cancer.

PHF8 is a JmjC domain-containing protein and erases repressive histone marks including H4K20me1 and H3K9me1/2. It binds to H3K4me3, an active histone mark usually located at transcription start sites (TSSs), through its plant homeo-domain, and is thus recruited and enriched in gene promoters. PHF8 is involved in the development of several types of cancer, including leukemia, prostate cancer, and esophageal squamous cell carcinoma. Herein we report that PHF8 is an oncogenic protein in human non-small cell lung cancer (NSCLC). PHF8 is up-regulated in human NSCLC tissues, and high PHF8 expression predicts poor survival. Our in vitro and in vivo evidence demonstrate that PHF8 regulates lung cancer cell proliferation and cellular transformation. We found that PHF8 knockdown induces DNA damage and apoptosis in lung cancer cells. PHF8 promotes miR-21 expression in human lung cancer, and miR-21 knockdown blocks the effects of PHF8 on proliferation and apoptosis of lung cancer cells. In summary, PHF8 promotes lung cancer cell growth and survival by regulating miR-21.

Quantitative assessment of the influence of common variations on 6p21 and lung cancer risk.

Genome-wide association studies have identified 6p21 as a new lung cancer susceptibility locus in populations of European descent. Since then, the relationship between common variations on 6p21 and lung cancer has been reported in various ethnic groups; however, these studies have yielded inconsistent results. To investigate this inconsistency, we performed a meta-analysis of 11 studies involving a total of 36,890 cases and 52,767 controls for three widely studies polymorphisms (rs4324798, rs3117582, and rs9295740) to evaluate its effect on genetic susceptibility for lung cancer. An overall random-effects per-allele odds ratio (OR) of 1.11 (95% confidence interval (CI) 1.04­1.19; P = 0.002) and 1.20 (95% CI 1.14­1.26; P < 10(−5)) was found for the rs4324798 and rs3117582 polymorphism, respectively. Marginal significant associations were also detected for rs9295740 with per-allele OR of 1.09 (95% CI 1.01­1.18; P = 0.03). In the subgroup analysis by ethnicity, significantly increased risks were found for the three polymorphisms among Caucasians. Similar results were also observed using dominant or recessive genetic models. This meta-analysis demonstrated that the three common variations (rs4324798, rs3117582, and rs9295740) on 6p21 are risk factors associated with increased lung cancer susceptibility, but these associations vary in different ethnic populations.

[Selective three-field lymphadenectomy for thoracic esophageal squamous carcinoma].

OBJECTIVE: To investigate the lymph node metastasis and the rational lymphadenectomy in thoracic esophageal carcinoma. METHODS: Eighty-seven patients with thoracic esophageal squamous carcinoma received esophagectomy plus two-field or three-field lymphadenectomy based on cervical ultrasonography. RESULTS: Thirty-five patients (40.2% ) with enlarged cervical nodes revealed by cervical ultrasonography received cervical lymphadenectomy. The proportion of cervical lymphadenectomy was 66.7% (16/24) in upper thoracic esophageal carcinomas, significantly higher than 30.2% (19/63) in middle and lower esophageal carcinomas (P=0.002). Regional and cervical lymph node metastasis were found in 48(55.2% ) and 17(19.5% ) patients respectively. The regional lymph node metastatic rates were 37.5% (9/24), 62.3% (33/53) and 60.0% (6/10) respectively in the patients with upper, middle, and lower thoracic esophageal carcinoma. The cervical lymph node metastatic rates in the patients with or without regional lymph node metastasis were 31.3% (15/48) and 5.1% (2/39) respectively(P=0.002). The rates of upper, mid, lower mediastinal and upper abdominal lymph node metastasis were 25.3%, 23.0%, 5.7%, and 24.1% respectively. Cervical lymph node metastasis was significantly correlated with upper and mid mediastinal metastasis (both P< 0.01), but not with lower mediastinal and upper abdominal lymph node metastasis. The overall postoperative morbidity rate was significantly higher in three field lymphadenectomy group than that in two field group(60.0% vs. 34.6%, P=0.020). CONCLUSION: Selective 3-field lymphadenectomy based on cervical ultrasonography should be performed in thoracic esophageal carcinoma, especially with upper and mid mediastinal lymph node metastasis.