[Effect of TLK2 Expression Regulated by MiR-21 on Proliferation and Apoptosis of Acute Myeloid Leukemia Cells].

OBJECTIVE: To investigate the effect of TLK2 expression regulated by miR-21 on proliferation and apoptosis of acute myeloid leukemia cells. METHODS: Seventy patients with AML admitted to our hospital from January 2019 to July 2022 were selected, while 30 patients with iron deficiency anemia were selected as the control group. Bone marrow mononuclear cells (BMMNCs) of the patients were obtained using Ficoll density gradient centrifugation. RT-qPCR was used to determine the expression levels of miR-21 and TLK2 mRNA in BMMNCs. Mimics-miR-21, mimics-NC, inhibitor-miR-21, inhibitor-NC and NC were transfected into HL-60 cells using liposome-mediated transfection technology. CCK-8 method was used to determine the activity of transfected HL-60 cells after treatment with cytarabine. The apoptosis rate of HL-60 transfected cells was determined by TUNEL method. The expression of TLK2 mRNA in HL-60 cells transfected with inhibitor-miR-21 was determined by RT-qPCR. RESULTS: The relative expression levels of miR-21 and TLK2 mRNA in BMMNCs of AML patients were significantly higher than those of controls (both P < 0.05). After HL-60 cells were treated with cytarabine, both the cell activity of inhibitor-miR-21 group and mimics-miR-21 group decreased significantly with the increase of cytarabine concentration (both P < 0.05). However, at each concentration point of cytarabine, the cell activity of inhibitor-miR-21 group was lower than that of control group (P < 0.05), while mimics-miR-21 group was higher than control group (P < 0.05). After HL-60 cells were treated with cytarabine, the apoptosis rate of inhibitor-miR-21 group was significantly increased (P < 0.05), while that of mimics-miR-21 group was significantly decreased (P < 0.05). After HL-60 cells were treated with inhibitor-miR-21, the relative expression of TLK2 mRNA decreased significantly (P < 0.05). CONCLUSION: miR-21 is highly expressed in AML patients, which may promote the apoptosis of AML cells by inhibiting the expression of TLK2.

Network pharmacology and molecular docking reveal the mechanisms of curcumin activity against esophageal squamous cell carcinoma.

Background: Curcumin (CUR), an effective traditional Chinese medicinal extract, displays good anti-cancer activity against various cancers. Nevertheless, the impacts and fundamental mechanisms of CUR to treat esophageal squamous cell carcinoma (ESCC) yet to be comprehensively clarified. This study examined the suppressive impacts of CUR on ESCC. Methods: For a comprehensive understanding of the effect of CUR in ESCC. The CUR targets and ESCC-related genes were identified respectively, and the intersection targets between CUR and ESCC were acquired. Then, we examined the intersection targets and discovered genes that were expressed differently in ESCC. Using DAVID, enrichment analyses were conducted on the targets of CUR-ESCC. The STRING database and Cytoscape v.3.9.1 were utilized to build networks for protein-protein interaction (PPI) and drug-target-pathway. Furthermore, the interactions between CUR and its core targets were confirmed by molecular docking studies. To confirm the effects of CUR on ESCC cells, in vitro experiments were finally conducted. Results: Overall, 47 potential CUR targets for ESCC treatment were identified. The KEGG pathway enrichment analysis identified 61 signaling pathways, primarily associated with the FoxO signaling, the cell cycle, cellular senescence, the IL-17 signaling pathway which play important roles in ESCC progression. In the PPI network and the docking results identified CHEK1 and CDK6 as the core targets that positively associated with ESCC survival. CUR arrested ESCC cells at the G2/M and S phases, as shown by flow cytometry. Colony formation and CCK8 assays showed that CUR can inhibit the proliferative ability of ESCC cells. The Transwell invasion results validated that CUR can significantly inhibit the invasion rates of ESCC cells. Conclusion: Collectively, these findings indicate that CUR exhibits pharmacological effects on multiple targets and pathways in ESCC.

Identification of anti-TMV active flavonoid glycosides and their mode of action on virus particles from Clematis lasiandra Maxim.

BACKGROUND: Tobacco mosaic virus (TMV) is a disreputable plant pathogen that causes a decline in the quality and yield of various economic crops. Natural products are important potential sources of biopesticides to control TMV. This study focuses on the discovery of anti-TMV active flavonoid glycosides and their mode of action on TMV particles from Clematis lasiandra Maxim. RESULTS: A new benzoyl acylated flavonoid glycoside, kaempferol 3-O-(2''-benzoyl)-ß-d-glucopyranosyl-7-O-α-l-rhamnopyranoside (1), and nine known flavonoids (2-10) were identified first from C. lasiandra. The hydroxyl group at C-7, E-p-coumarate at C-6'' in the Glc of C-6, and the glucuronic acid at C-3 were functional groups for the antiviral flavonoid glycosides. Flavonoids 2, 5, and 6 showed higher inactivation efficacies of 64.62% to 82.54% compared with ningnanmycin at 500 µg ml-1 . The protective and curative efficacies for 2 and 5 were 57.44-59.00% and 41.17-43.92% at 500 µg ml-1 , respectively. Compound 5 showed higher TMV systemic resistance with control efficacies of 41.64%, 36.56% and 27.62% at concentrations of 500, 250 and 125 µg ml-1 compared with ningnanmycin in K326 tobaccos, respectively. Compound 5 can directly fracture TMV particles into small fragments combining with the fusion phenomena, and TMV-CP was an important target for 5 to break TMV particles. CONCLUSION: Flavonoid glycosides from C. lasiandra showed potent antiviral activities against TMV with multiple modes of action including inactivation, protective and curative effects, and inducing systemic resistance. TMV-CP was an important target for active flavonoid glycosides to fracture TMV particles. The results provided evidence that flavonoid glycosides from C. lasiandra have the potential to control TMV.

Treatment Effect of Tuftsin and Antigen Peptide Combined with Immune Cells on Colorectal Cancer.

BACKGROUND The aim of this study was to investigate the effect of antigenic peptides on dendritic cell maturation and activation as well as the role of dendritic cell induced cell function. The tumor-specific cytotoxic T lymphocytes induced by activation of the dendritic cells were also evaluated. MATERIAL AND METHODS SW-480 cell lysate and peptide antigens were selected as adjuvants in dendritic cell sensitization, and tuftsin was used to induce the phagocytosis of dendritic cells. Immature dendritic cells were stimulated with the antigen and adjuvant as follows: group A was negative control; group B was SW-480 (20 µg/mL); group C was SW-480 (20 µg/mL)+tumor necrosis factor (TNF)-alpha (10 µg/mL); group D was SW-480 (20 µg/mL)+tuftsin (20 µg/mL); group E was antigen peptide (2 µg/mL); group F was antigen peptide (2 µg/mL)+TNF-alpha (10 µg/mL); group G was antigen peptide (2 µg/mL)+tuftsin (20 µg/mL). Cytotoxic T lymphocytes activation and in vitro anti-tumor effects were examined by detecting the maturation marks of dendritic cells as well as interleukin (IL)-10 and IL-12 levels secreted by dendritic cells. Cells with the strongest immunizing effects were injected into nude mice and tumor suppression status was evaluated. RESULTS Group D (SW-480+tuftsin), group E (antigen peptides), group F (antigen peptide+TNF-alpha), and group G (antigen peptides+tuftsin) displayed significant differences compared to the control group (P<0.05). Group G (antigen peptides+tuftsin) could also promote the secretion of cytokines IL-12, as well as inhibit cytokine IL-10 secretion, compared to the other experimental groups (P<0.05). In the in vivo experiments of tumor inhibitions, antigenic polypeptide+tuftsin was the most effective (P<0.05). CONCLUSIONS Combination of cytotoxic T lymphocytes and T peptide therapy in treating human colorectal cancer might be used as a new treatment strategy based on adoptive cellular immunotherapy.

Glutamate dehydrogenase inhibits tumor growth in gastric cancer through the Notch signaling pathway.

Glutamate dehydrogenase (GDH) is a key enzyme in glutaminolysis and can regulate allosteric functions. Immunohistochemical study found that GDH expressed in gastric cancer cell cytoplasm and membrane, and a few located in the nucleus, ranging from light yellow to tan to sepia. According to the analysis by Kaplan Meier survival curve and the Log-Rank test, the median survival of GDH high expression in patients was 51.7 months with 95% confidence intervals (CI) was 41.138-55.262. The expression level of GDH was significantly reduced after silencing GDH gene in gastric cancer cells and tissues. Further, after silencing GDH gene, gastric cancer cell migration and invasion ability were decreased significantly. Protein expression of. In addition, tumor growth was significantly reduced after silencing GDH gene. In vivo and in vitro experiments suggest that GDH can decrease gastric cancer cell migration and invasion, thus inhibiting tumor growth.

Enhanced antitumor effect of cytotoxic T lymphocytes induced by dendritic cells pulsed with colorectal cancer cell lysate expressing α-Gal epitopes.

Colorectal cancer (CRC) is one of the most common types of gastrointestinal malignancy. Traditional therapeutic options for CRC exhibit a limited effect. Adoptive cellular therapy has emerged as a new treatment strategy for CRC. Dendritic cells (DCs) are potent antigen-presenting cells. Specific cytotoxic T lymphocytes (CTLs) activated by DCs pulsed with tumor lysate have been reported to be a safe and promising treatment approach for CRC. However, the antitumor effect of specific CTLs remains limited. The low immunogenicity of tumor-associated antigens (TAAs) is the main reason for this limited therapeutic effect. In the present study, α-gal epitopes were synthesized on the CRC cell line SW620 to increase the immunogenicity of TAAs. DCs were pulsed with α-gal-expressing tumor lysate and CTLs were activated by these DCs. The cytotoxicity of CTLs was measured in vitro. The results demonstrated that DCs pulsed with α-gal-expressing tumor lysate can increase the frequency of CD3+CD8+ CTLs and natural killer T cells, increase the level of tumor necrosis factor-α produced by CTLs and enhance the cytotoxicity of CTLs against tumor cells. Therefore, this novel approach may be an effective treatment strategy for patients with CRC.

Application of highly efficient and lowly toxic bufadienolides screened from toad skin in lymphatic chemotherapy for colorectal cancer through a lymphatic metastatic model.

BACKGROUND: Lymph node metastasis (LNM) remains a major obstacle to treat colorectal cancer (CRC). Increasing evidences have suggested that bufadienolides contain several fractions displaying antitumor activity and may be applied in lymphatic chemotherapy. However, effects of the highly efficient and lowly toxic (HELT) bufadienolides on CRC in lymphatic chemotherapy have not been reported. METHODS: Adenosine triphosphate tumor chemosensitivity assays (ATP-TCA) was performed to detect the inhibition rate (IR) of fractions of bufadienolides to cytokine-induced killer (CIK) cells and tumor cells. HELT fraction-loaded emulsions of different concentrations were prepared. Nude mouse bearing HCT116 tumors in footpad received high-dose emulsion (HD-E), middle-dose emulsion (MD-E), low-dose emulsion (LD-E), control emulsion (CE), Cinobufacini Injection (CI), or normal saline (NS), respectively. Hematoxylin and eosin (H&E) staining, Flow Cytometry (FCM), enzyme-linked immune sorbent assay (ELISA) and hematological examination were applied to evaluate therapeutic effects and potential toxicity. RESULTS: F18 and F19 were screened out as HELT fractions in vivo and F18-loaded emulsions of different concentrations for lymphatic administration were prepared. We confirmed that HD-E and MD-E produced obvious antitumor activities in footpad tumors and LNM compared with other groups in vitro. We also verified the effects of F18-loaded emulsions on activating hematopoietic function, stimulating proliferation of the spleen and natural killer (NK) cells, and promoting the secretion of IFN-γ and IgG1, although HD-E performed mild toxicity on liver. CONCLUSION: The present study demonstrated that lymphatic chemotherapy with HELT fraction of bufadienolides could be an effective approach to the treatment of CRC patients with LNM.

Expression of CUEDC2 in colorectal cancer with different invasion and migration abilities.

OBJECTIVE: To investigate CUE domain containing 2 ( CUEDC2) expression in colorectal cancer with different invasion and migration abilities. METHODS: Fresh colon cancer tissues, obtained from patients with or without lymph node metastasis who were treated at the Department of General Surgery, Chinese People's Liberation Army General Hospital, and SW620 and HT29 colorectal cancer cell lines, were analysed for CUEDC2 expression. RESULTS: Real-time polymerase chain reaction showed significantly higher CUEDC2 mRNA levels in colon cancer tissue from patients with ( n = 8) versus without ( n = 8) lymph node metastasis, and in SW620 versus HT29 cells. Western blots revealed significantly higher CUEDC2 protein levels in colon cancer tissues from patients with versus without lymph node metastasis, and in SW620 versus HT29 cells. Colorectal cancer tissues from patients with lymph node metastasis showed more intense immunohistochemical staining and moderate staining of cell nuclei and cytoplasm versus less intense/weak staining in tissues from patients without lymph node metastasis. CONCLUSIONS: CUEDC2 is highly expressed in colorectal cancer tissues and colorectal cancer cell lines with high invasion and migration ability. CUEDC2 may be involved in promoting invasion and metastasis in colorectal cancer.

[Speciation and Transformation of Phosphorus in Sediments During the Redox Cycle].

To study the mechanism of phosphorus cycling in sediment during the redox cycle, changes in physicochemical properties of overlying water and various forms of phosphorus in sediments were investigated as a way to quantify the redistribution of phosphorus. Additionally, the effect of the release flux of phosphate from sediments under controlled redox conditions was analyzed. The results showed that the redox potential Eh and the pH system, sulfur system, carbon system, and iron-related changes exhibited periodicity and played an important role in explaining the migration and transformation mechanism in the interface phosphorus of the sediment-water phase. During the redox cycle, the phosphorus content of each species varied with the redox conditions and time. Because of this, quantitative analysis based on changes in water-sediment phosphorus could be obtained. Reducible phosphorus (BD-P) and iron-aluminum-bound phosphorus (NaOH-rP) were reversibly redistributed into weakly adsorbed phosphorus (NH4Cl-P), polyphosphorus/organophosphorous (NaOH-nrP), residual phosphorus (Rest-P), and interstitial water-soluble active phosphorus (SRP). Additionally, 93.7% of phosphorus in the sediment was not released into the water phase during the reduction reaction. The 92% of change in the overlying water total phosphorus (TP) was the SRP of overlying water, which showed that the exchange of the sediment-water phase were mainly soluble active phosphorus in this cycle. According to Fick's First Law, the maximum phosphorus flux was 0.58 mg·(m2·d)-1 during reduction and 0.16-0.22 mg·(m2·d)-1 on day seven of the oxidation phase. In the oxidation stage, the diffusion flux decreased with time, while the opposite trend occurred in the reduction reaction. This indicated that the anaerobic state accelerated the diffusion of phosphorus in sediments, and that oxygen exposure decreased the phosphorus flux in sediments.

Improving immunotherapy for colorectal cancer using dendritic cells combined with anti-programmed death-ligand in vitro.

Monoclonal antibodies recognizing programmed death-ligand 1 (PD-L1) have been used for the clinical treatment of diverse tumor types as a form of immune checkpoint inhibitor, with a favorable therapeutic effect. Dendritic cells (DCs) are potent antigen-presenting cells that serve a pivotal role in the activation of T cells, particularly cytotoxic T lymphocytes (CTLs). DC vaccines loaded with tumor antigens, DC-CTLs and activated T cells have been revealed to be a safe and effective treatment approach against colorectal cancer within a clinical setting. In addition to tumor cells, PD-L1 is also highly expressed on DCs. As research examining the association between anti-PD-L1 and DCs is lacking, the present study compared the expression of PD-L1 on DCs in the peripheral blood of healthy donors and patients with colorectal cancer. Following the application of anti-PD-L1, the DC phenotypes, function of DC-mediated T cell induction and the cytotoxicity of CTLs were investigated by flow cytometry. The present study revealed that treatment with anti-PD-L1 may promote the maturation of DCs and enhance the functionality of the DC1 subtype. It may also increase the number of CTLs that are activated and produce CTL cells with more potent anti-tumor activity. Therefore, the creation of DC vaccines in conjunction with anti-PD-L1 may be an effective future treatment strategy for patients with colorectal cancer.

Accumulation and suppressive function of regulatory T cells in malignant ascites: Reducing their suppressive function using arsenic trioxide in vitro.

Although adoptive cell therapy (ACT) has demonstrated effective and remarkable clinical responses in several studies, this approach does not lead to objective clinical responses in all cases. The function of ACT is often compromised by various tumor escape mechanisms, including the accumulation of immunoregulatory cells. As a result of peritoneal metastasis in the terminal stage, malignant ascites fluid lacks effectiveness and is a poor prognostic factor for gastric cancer. The present study assessed T-cell subsets in lymphocytes derived from malignant ascites, and investigated the effects of arsenic trioxide (As2O3) on regulatory T cells (Tregs) and ascites-derived tumor-infiltrating lymphocytes (TILs) in vitro. In this study, lymphocytes were separated from malignant ascites and T-cell subsets were detected via flow cytometry. Forkhead box P3 (FoxP3) expression was assessed by immunohistochemistry and reverse transcription-quantitative polymerase chain reaction. In addition, cytokines, including interleukin-10 (IL-10), transforming growth factor-ß (TGF-ß), and interferon-γ (IFN-γ), were measured by enzyme-linked immunosorbent assay (ELISA). Abundant Tregs were observed in ascites lymphocytes, which and exhibited a significantly increased frequency compared with that in the peripheral blood of patients. Furthermore, As2O3 treatment significantly reduced Treg numbers and Foxp3 mRNA levels in vitro (P<0.05). IFN-γ levels in the supernatant of ascites-derived TILs were increased by As2O3, whereas IL-10 and TGF-ß levels were significantly reduced (P<0.05). As2O3 may induce selective depletion and inhibit immunosuppressive function of Tregs, and may enhance the cytotoxic activity of ascites-derived TILs.

Prediction of rupture risk in anterior communicating artery aneurysms with a feed-forward artificial neural network.

OBJECTIVES: Anterior communicating artery (ACOM) aneurysms are the most common intracranial aneurysms, and predicting their rupture risk is challenging. We aimed to predict this risk using a two-layer feed-forward artificial neural network (ANN). MATERIALS AND METHOD: 594 ACOM aneurysms, 54 unruptured and 540 ruptured, were reviewed. A two-layer feed-forward ANN was designed for ACOM aneurysm rupture-risk analysis. To improve ANN efficiency, an adaptive synthetic (ADASYN) sampling approach was applied to generate more synthetic data for unruptured aneurysms. Seventeen parameters (13 morphological parameters of ACOM aneurysm measured from these patients' CT angiography (CTA) images, two demographic factors, and hypertension and smoking histories) were adopted as ANN input. RESULTS: Age, vessel size, aneurysm height, perpendicular height, aneurysm neck size, aspect ratio, size ratio, aneurysm angle, vessel angle, aneurysm projection, A1 segment configuration, aneurysm lobulations and hypertension were significantly different between the ruptured and unruptured groups. Areas under the ROC curve for training, validating, testing and overall data sets were 0.953, 0.937, 0.928 and 0.950, respectively. Overall prediction accuracy for raw 594 samples was 94.8 %. CONCLUSION: This ANN presents good performance and offers a valuable tool for prediction of rupture risk in ACOM aneurysms, which may facilitate management of unruptured ACOM aneurysms. KEY POINTS: • A feed-forward ANN was designed for the prediction of rupture risk in ACOM aneurysms. • Two demographic parameters, 13 morphological aneurysm parameters, and hypertension/smoking history were acquired. • An ADASYN sampling approach was used to improve ANN quality. • Overall prediction accuracy of 94.8 % for the raw samples was achieved.

Ultrasound-guided percutaneous microwave ablation vs. surgical resection for thoracoabdominal wall implants from hepatocellular carcinoma: intermediate-term results.

OBJECTIVE: The aims of this study were to compare the clinical outcomes between ultrasound (US)-guided percutaneous microwave ablation (MWA) and surgical resection (SR) in patients with thoracoabdominal wall implants from hepatocellular carcinom (HCC) and to identify the prognostic factors associated with the two treatment methods. MATERIALS AND METHODS: A total of 47 patients (mean age, 56.7 ± 15.9 years, range, 18-78 years; 34 men and 13 women) with 61 thoracoabdominal wall HCC seeding were included from April 2007 to May 2017. Twenty-five patients underwent US-guided MWA and 22 patients underwent SR. Survival, recurrence and liver function were compared between the two groups. Effect of changes in key parameters (i.e. overall survival (OS), disease-free survival (DFS) and local tumour reoccurrence-free (LTRF)) was statistically analysed with the log-rank test. Univariate and multivariate analyses were performed on several clinicopathological variables to identify factors affecting long-term outcome and recurrence. RESULTS: The OS, DFS and LTRF after MWA were comparable to those of SR (p =0.493, p = 0.578 and p =0.270, respectively). Estimated 5-year overall survival rates were 63% after MWA and 48.1% after SR; for disease-free survival, estimated 5-year rates were 67.5% after MWA and 48.8% after SR; estimated 24-month LTRF rates were 71.3% after MWA and 87.8% after SR. The MWA group had less surgical time (p = <0.001), estimated blood loss (p = <0.001) and post-operative hospitalisation (p = 0.032) and cost (p = 0.015). Multivariate analysis showed remnant intrahepatic tumour (p =0.007), Child Pugh grade (p = 0.009) and metastasis (p= <0.001), were predictors for survival rate. CONCLUSIONS: Ultrasound-guided percutaneous MWA is a safe and effective treatment method for metastatic HCC on the thoracoabdominal wall with similar outcomes to SR. Residual intrahepatic HCC, Child Pugh grade and distant metastasis are predictors for survival.

[Increased expressions of peripheral PD-1+ lymphocytes and CD4+CD25+FOXP3+ T cells in gastric adenocarcinoma patients].

Objective To detect the frequencies of peripheral programmed death-1+ (PD-1+) lymphocytes and CD4+CD25+FOXP3+ regulatory T cells in patients with gastric adenocarcinoma. Methods The study enrolled 29 patients with gastric adenocarcinoma and 29 age- and sex-matched healthy controls. Frequencies of PD-1+ lymphocytes and CD4+CD25+FOXP3+ regulatory T cells were detected using flow cytometry. Results The number of PD-1+ lymphocytes and CD4+CD25+FOXP3+ regulatory T cells in peripheral blood was higher in patients with gastric adenocarcinoma than that in the control group. Moreover, linear correlation analysis indicated a positive correlation between PD-1 expression and frequency of CD4+CD25+FOXP3+ regulatory T cells in peripheral blood of the patients. Conclusion Gastric adenocarcinoma patients present with increased PD-1+ lymphocytes and CD4+CD25+FOXP3+ regulatory T cells in the peripheral blood.

[Value of the preoperative TNM staging and the longest tumor diameter measurement of gastric cancer evaluated by MSCT].

OBJECTIVE: To explore the value of MSCT in the preoperative TNM staging and the longest tumor diameter measurement (RESIST standard) of gastric cancer. METHODS: Clinical data of 153 consecutive patients with biopsy-confirmed gastric carcinoma who were preoperatively evaluated with enhanced MSCT scanning in our hospital from January 2012 to March 2013 were retrospectively analyzed. Consistency comparison was performed between preoperative TNM staging and the longest tumor diameter measurement and histopathological findings. RESULTS: T-staging consistency of Kappa value was 0.566, and accuracy was 71.2%. N-staging consistency of Kappa value was 0.284, and accuracy was 47.7%. The Kappa value of M-staging consistency was 0.893, and accuracy was 98.7%. The overall accuracy of TNM staging consistency was 66.7% (102/153) with a Kappa value of 0.573. Effective measurement of the longest cancer diameter was carried out in 53 patients. There was no significant difference between preoperative longest tumor diameter acquired by MSCT and postoperative tumor measurement [(68.8 ± 40.6) mm vs. (64.2 ± 36.2) mm, P=0.969]. CONCLUSION: MSCT is accurate in preoperative TNM staging and longest tumor diameter measurement of gastric cancer compared with postoperative pathological examination, and can provide reliable evidence for preoperative staging and neoadjuvant therapy evaluation of gastric cancer, but it is unfavorable to evaluate the lymph node metastasis.