miR-29b-3p suppresses the malignant biological behaviors of AML cells via inhibiting NF-κB and JAK/STAT signaling pathways by targeting HuR.

BACKGROUND: HuR/ELAVL1 (embryonic lethal abnormal vision 1) was a downstream target of miR-29b in some cancer cells. HuR protein exerts important prognostic effects of involving in the pathogenesis and development of acute myeloid leukemia (AML). This study aims to investigate the role of miR-29b-3p in biological behaviors of AML cells by targeting HuR and the involvement of the NF-κB and JAK/STAT signaling pathways. METHODS: The expressions of HuR and miR-29b-3p in AML cells were determined using RT-qPCR and Western blot, and the association between them was analyzed using the Spearman method. Next, the target relationship between HuR and miR-29b-3p was predicted by biological information databases and verified by the dual-luciferase reporter gene assay. MTS, methyl cellulose, flow cytometry and transwell assay were employed to detect the cell proliferation, clone formation, cell cycle and apoptosis, invasion and migration respectively, the effect of miR-29b-3p targeted HuR on the biological behaviors of AML cells was explored after over- /down-expression of miR-29b-3p and rescue experiment. Then, immunofluorescence assay and western blot were employed to detect location expression and phosphorylation levels of NF-κB and JAK/STAT signaling pathways related molecules respectively. RESULTS: HuR was negatively correlated with miR-29b-3p, and was the downstream target of miR-29b-3p in AML cells. When miR-29b-3p was overexpressed in AML cells, HuR was down-regulated, accompanied by cell viability decreased, cell cycle arrest, apoptosis increased, invasion and migration weakened. Moreover, the opposite result appeared after miR-29b-3p was down-regulated. The rescue experiment showed that miR-29b-3p inhibitor could reverse the biological effect of HuR down-regulation in AML cells. Molecular pathway results showed that miR-29b-3p could inhibit p65 expression in nucleus and phosphorylation levels of p65, IκBα, STAT1, STAT3 and STAT5. CONCLUSION: miR-29b-3p can inhibit malignant biological behaviors of AML cells via the inactivation of the NF-κB and JAK/STAT signaling pathways by targeting HuR. miR-29b-3p and its target HuR can be used as a new potential molecular for AML treatment.

Comparative analysis of leaching residual monomer and biological effects of four types of conventional and CAD/CAM dental polymers: an in vitro study.

OBJECTIVES: The objective of this study is to investigate leaching residual monomer and biological effects of four types of conventional and computer-aided design/computer-aided manufacturing (CAD/CAM) dental polymers on human gingival fibroblasts (HGFs). MATERIALS AND METHODS: A total of 540 disk-shaped specimens were fabricated from four different materials (n=135 per group): compression-molding polymethylmethacrylate (PMMA) (conventional denture polymer), CAD/CAM PMMA (CAD/CAM denture polymer), bis-acrylic composite resin (conventional temporary polymer), and CAD/CAM PMMA (CAD/CAM temporary polymer). Specimens were eluted in cell culture medium for 72 h at 37°C, and the residual monomer in eluates subsequently was measured by high-performance liquid chromatography (HPLC). The biological effects of material eluates on HGFs were analyzed by CCK-8 assay, flow cytometry, real-time quantitative PCR, Western blotting, and enzyme-linked immunosorbent assay (ELISA) to identify cell death patterns and its biological mechanism. RESULTS: Methyl methacrylate (MMA) was detected only in compression-molding PMMA, and by-products were detected in bis-acrylic composite resin. The cell proliferation of CAD/CAM denture polymer or CAD/CAM temporary polymer was greater than that of compression-molding PMMA or bis-acrylic composite resin at 72 h in culture. No apoptosis and necrosis were detected in CAD/CAM dental polymers. Apoptosis was detected only in bis-acrylic composite resin and further confirmed by the upregulation of Bax and cleaved Caspase-3, as well as the downregulation of Bcl-2 gene. And no significant variation in inflammatory cytokines secretion was observed in all materials. CONCLUSIONS: CAD/CAM dental polymers (including temporary and denture polymers) have favorable biocompatibility due to lower residual monomer, which provides scientific evidence to the controversy of biocompatibility of conventional and CAD/CAM dental polymers. CLINICAL RELEVANCE: The use of CAD/CAM dental polymers is recommended in the fabrication of temporary restorations and dentures due to their favorable biocompatibility.

Two Tellurium(IV)-Based Sulfates Exhibiting Strong Second Harmonic Generation and Moderate Birefringence as Promising Ultraviolet Nonlinear Optical Materials.

Two tellurium(IV)-based sulfate nonlinear optical (NLO) materials, Te2O3(SO4) and Te(OH)3(SO4)·H3O, were successfully synthesized via the mild hydrothermal method. Te2O3SO4 has a two-dimensional (2D) structure consisting of [Te6O12]∞ layers as well as [SO4] groups. Te(OH)3(SO4)·H3O features a simple 0D structure made up of an isolated [TeO3] pyramid and a [SO4] tetrahedra. Both of them are phase-matching materials and show remarkable powder second harmonic generation (SHG) efficiencies about 6 and 3 times that of KH2PO4 (KDP), respectively, for Te2O3SO4 and Te(OH)3(SO4)·H3O. Especially for Te(OH)3(SO4)·H3O, in addition to a large SHG response, it possesses a short UV cutoff edge (∼233 nm) as well as moderate birefringence (0.052@546.1 nm). Furthermore, theoretical calculations confirmed that their strong SHG effects are due to the synergistic effect of the [TeO3] pyramid and [SO4] tetrahedra.

Silencing of the CrkL gene reverses the doxorubicin resistance of K562/ADR cells through regulating PI3K/Akt/MRP1 signaling.

BACKGROUND: Doxorubicin is a first-line chemotherapy agent on human myelogenous leukemia clinical treatment, but the development of chemoresistance has largely limited curative effect. In this study, we aimed to evaluate the biological function and molecular mechanisms of CrkL to Doxorubicin resistance. METHODS: Quantitative reverse transcription-PCR (qRT-PCR) assay was performed to examine the expression of CrkL in K562 and K562/ADR cells. The expression of CrkL was silenced through RNA interference technology. MTT assay and flow cytometry were performed to detect the proliferation inhibition and apoptosis rate after CrkL siRNA transfection. The protein expression changes of PI3K/AKT/MRP1 pathway induced by CrkL siRNA were observed by Western Blot assay. Xenograft tumor model was carried out to observe tumor growth in vivo. RESULTS: We observed that silencing of CrkL could effectively increase apoptosis rate induced by doxorubicin and dramatically reversed doxorubicin resistance in K562/ADR cells. Further studies revealed knockdown CrkL expression suppressed PI3K/Akt/MRP1 signaling, which indicated CrkL siRNA reversed doxorubicin effect through regulating PI3K/Akt/MRP1 pathway. In addition, overexpression of MRP1 could evidently reduce apoptosis rate and reversed the inhibitory effects of doxorubicin resistance caused by CrkL siRNA on K562/ADR cells. Finally, in vivo experiments revealed that CrkL silencing acted a tumor-suppressing role in myelogenous leukemia via regulating PI3K/Akt/MRP1 signaling. CONCLUSION: Together, we indicated that CrkL is up-regulated in myelogenous leukemia cells and silencing of CrkL could reverse Doxorubicin resistance effectively. These results show a potential novel strategy for intervention chemoresistance in myelogenous leukemia during chemotherapy.

E35 ablates acute leukemia stem and progenitor cells in vitro and in vivo.

Leukemia stem cells (LSCs) have critical functions in acute leukemia (AL) pathogenesis, participating in its initiation and relapse. Thus, identifying new molecules to eradicate LSCs represents a high priority for AL management. This work identified E35, a novel Emodin derivative, which strongly inhibited growth and enhanced apoptosis of AL stem cell lines, and primary stem and progenitor cells from AL cases, while sparing normal hematopoietic cells. Furthermore, functional assays in cultured cells and animals suggested that E35 preferentially ablated primitive leukemia cell populations without impairing their normal counterparts. Moreover, molecular studies showed that E35 remarkably downregulated drug-resistant gene and dramatically inhibited the Akt/mammalian target of rapamycin signaling pathway. Notably, the in vivo anti-LSC activity of E35 was further confirmed in murine xenotransplantation models. Collectively, these findings indicate E35 constitutes a novel therapeutic candidate for AL, potentially targeting leukemia stem and progenitor cells.

Explorations of Second-Order Nonlinear Optical Materials in the Alkaline Earth Barbiturate System: Noncentrosymmetric Ca(H3C4N2O3)2·H2O and Centrosymmetric Sr(H5C8N4O5)2·4H2O.

Two new alkaline earth barbiturates, Ca(H3C4N2O3)2·H2O and Sr(H5C8N4O5)2·4H2O, were synthesized via the mild hydrothermal technique. For Ca(H3C4N2O3)2·H2O, the stacking (H3C4N2O3)- anions along the c axis are interconnected by CaO7 polydedra forming a three-dimensional structure. Also, for Sr(H5C8N4O5)2·4H2O, it has two-dimensional layers composed by SrO7 polyhedra and (H5C8N4O5)- anions. The (H5C8N4O5)- anions can be seen as two (H3C4N2O3)- anions connected each other via the N-C bond. Powder second-harmonic generation (SHG) measurements revealed that Ca(H3C4N2O3)2·H2O is a phase-matchable material with a moderate SHG response (ca. 1.15× that of KDP). Furthermore, the birefringence values of the two crystals were measured as 0.49 and 0.475 at 546.1 nm, respectively. The theoretical calculations showed that the SHG response and large birefringence were primarily caused by the (H3C4N2O3)- and (H5C8N4O5)- groups.

A(H3C3N3O3)(NO3) (A = K, Rb): Alkali-Metal Nitrate Isocyanurates with Strong Optical Anisotropy.

The first alkali-metal nitrate isocyanurates, A(H3C3N3O3)(NO3) (A = K, Rb), were synthesized by the tactic of introducing (NO3)- into isocyanurate with a mild hydrothermal technique. They crystallized into the same monoclinic centrosymmetric (CS) space group P21/c, which featured a 2D [(H3C3N3O3)(NO3)]∞ layered structure separated by K+ and Rb+ cations, respectively. Both compounds exhibited short ultraviolet cutoff edges (λcutoff = 228 and 229 nm) and large birefringences (Δn = 0.253 and 0.224 at 546.1 nm). More importantly, in comparison with most of the isocyanurates and nitrates, they have better thermal stability with decomposition temperatures up to 319.8 and 324.4 °C. In addition, our theoretical calculations reveal that the π-conjugated groups play significant roles in improving the optical anisotropy. Remarkably, introducing a π-conjugated inorganic acid radical (NO3)- into isocyanurate is an extremely meaningful strategy to explore new UV birefringent crystals.

Rational Design of the Metal-Free KBe2 BO3 F2 ⋠(KBBF) Family Member C(NH2 )3 SO3 F with Ultraviolet Optical Nonlinearity.

The first fluorosulfonic ultraviolet (UV) nonlinear optical (NLO) material, C(NH2 )3 SO3 F, is rationally designed by taking KBe2 BO3 F2 (KBBF) as the parent compound. C(NH2 )3 SO3 F features similar topological layers as KBBF by replacing inorganic (BO3 )3- with organic C(NH2 )3 + trigonal units and BeO3 F with SO3 F- tetrahedra. Therefore, C(NH2 )3 SO3 F is a metal-free UV NLO crystal. Benefiting from the coplanar configuration of the C(NH2 )3 + cationic groups, it possesses a large SHG response of 5×KDP and moderate birefringence of 0.133@1064 nm. Besides, it has a short UV cutoff edge of 200 nm. The calculated results reveal the shortest SHG phase-matching wavelengths can reach 200 nm. These findings highlight the exploration of metal-free compounds as nontoxic and low-cost UV NLO materials as a new research area.

KLi(HC3N3O3)·2H2O: Solvent-drop Grinding Method toward the Hydro-isocyanurate Nonlinear Optical Crystal.

A novel mixed alkali hydro-isocyanurate, KLi(HC3N3O3)·2H2O was first prepared in AOH-BOH-H3C3N3O3 (A/B = Li/Na/K/Rb/Cs) system via a solvent-drop grinding method. KLi(HC3N3O3)·2H2O shows a large second harmonic generation response (5.3 × KH2PO4) with an ultraviolet cutoff edge of 237 nm. More importantly, the bulk single crystal can be readily grown through water solution technique. Characterization of these crystals indicates that KLi(HC3N3O3)·2H2O has a high laser damage threshold (LDT) (4.76 GW/cm2) and exhibits a large birefringence (Δ n = 0.186@514 nm), which reduces Type I phase-matching to 246 nm.

Metabolic characteristics revealing cell differentiation of nasopharyngeal carcinoma by combining NMR spectroscopy with Raman spectroscopy.

BACKGROUND: The staging system of nasopharyngeal carcinoma (NPC) has close relationship with the degree of cell differentiation, but most NPC patients remain undiagnosed until advanced phases. Novel metabolic markers need to be characterized to support diagnose at an early stage. METHODS: Metabolic characteristics of nasopharyngeal normal cell NP69 and two types of NPC cells, including CNE1 and CNE2 associated with high and low differentiation degrees were studied by combining 1H NMR spectroscopy with Raman spectroscopy. Statistical methods were also utilized to determine potential characteristic metabolites for monitoring differentiation progression. RESULTS: Metabolic profiles of NPC cells were significantly different according to differentiation degrees. Various characteristic metabolites responsible for different differentiated NPC cells were identified, and then disordered metabolic pathways were combed according to these metabolites. We found disordered pathways mainly included amino acids metabolisms like essential amino acids metabolisms, as well as altered lipid metabolism and TCA cycle, and abnormal energy metabolism. Thus our results provide evidence about close relationship between differentiation degrees of NPC cells and the levels of intracellular metabolites. Moreover, Raman spectrum analysis also provided complementary and confirmatory information about intracellular components in single living cells. Eight pathways were verified to that in NMR analysis, including amino acids metabolisms, inositol phosphate metabolism, and purine metabolism. CONCLUSIONS: Methodology of NMR-based metabolomics combining with Raman spectroscopy could be powerful and straightforward to reveal cell differentiation development and meanwhile lay the basis for experimental and clinical practice to monitor disease progression and therapeutic evaluation.

Inhibition of N822K T>A mutation-induced constitutive c-KIT activation in AML cells triggers apoptotic and autophagic pathways leading to death.

Background: The D816V mutation of c-KIT can constitutively activate tyrosine kinase, thereby promote core binding factor acute myeloid leukemia (CBF-AML) cell proliferation and inhibit apoptosis. Previous studies have indicated similar proliferation and apoptosis between N822K and D816V mutations.The current study aims to determine the occurrence and potential functions of N822K mutation-induced c-KIT activation in AML cells, and explore possible mechanisms of poor prognosis of CBF-AML. Methods: c-KIT N822K mutation status in AML cells was determined by exon 17 sequencing. The level of c-KIT expression was detected by flow cytometry (FCM) and colony formation was assessed after hu-SCF stimulation. After exposure to sunitinib (a kind of tyrosine kinase inhibitor, TKI), cell proliferation inhibition was tested by MTT, cell cycle and apoptosis were measured by FCM, autophagy was assessed by fluorescence microscopy and immunoblotting. Results: Kasumi-1 cell line was detected to bear c-KIT N822K (T>A) mutation. After hu-SCF stimulation, CD117 expression was decreased and the colony formation efficiency was not altered in Kasumi-1 cells. After sunitinib inhibited the c-KIT activity, the colony formation efficiency was reduced, and the half-maximal inhibitory concentration (IC50) of sunitinib was low (0.44±0.17µM) at 48 hours. Moreover, cells were arrested in G0/G1 phase, corresponding to an increase of apoptosis ratio. Acidic vesicular organelles (AVO) were observed along with an altered expression of autophagy-related proteins in Kasumi-1 cells. Conclusions: Our data indicated that inhibition of N822K T>A mutation-induced constitutive c-KIT activation in AML cells triggered apoptotic and autophagic pathways leading to death, and c-KIT N822K mutation may have clinical application as a CBF-AML treatment target.

Appraising Oncogenic Long Noncoding RNAs as Novel Biomarkers for Oral Tongue Squamous Cell Carcinoma: a Meta-Analysis.

BACKGROUND: Long noncoding RNAs (lncRNAs) are rapidly emerging as a novel class of transcripts associated with tumorigenesis and prognosis of oral tongue squamous cell carcinoma (OTSCC). The goal of this study was to perform a meta-analysis of the prognostic utility of tumorigenic lncRNAs as novel biomarkers of OTSCC and their associations with the clinicopathological features of OTSCC. METHODS: Online databases were searched for eligible studies. The hazard ratio (HR) with 95% CI was used to predict survival. The individual p-values for clinicopathological features were respectively combined using Fisher's method. RESULTS: A total of 11 studies covering 1,138 OTSCC cases were included for quantitative data synthesis. Oncogenic lncRNA expression signature was closely related to shortened overall survival (OS) of patients with OTSCC (HR = 1.85, 95% CI: 1.45 - 2.37, p = 0.001; I2 = 64.9%; p = 0.002). Stratified analysis based on clinicopathological features showed that abnormally expressed lncRNAs were significantly associated with clinical stage (p = 0.000), node metastasis (p = 0.025), T stage (p = 0.000), and nodal stage (p = 0.000). CONCLUSIONS: Collectively, oncogenic lncRNAs were implicated in the development and progression of OTSCC, and may serve as novel biomarkers for predicting the prognosis of OTSCC patients.

Emodin and Its Combination with Cytarabine Induce Apoptosis in Resistant Acute Myeloid Leukemia Cells in Vitro and in Vivo.

BACKGROUND/AIMS: Acute myeloid leukemia (AML) remains a hematologic malignancy with poor survival and a high risk of relapse, which is mainly caused by the emergence of multidrug resistance (MDR). The identification of novel agents to improve therapeutic strategies becomes important priority for AML treatment. It has been shown that emodin has therapeutic effects on many kinds of human malignant tumors. In this study, we investigated the anti-leukemia effects of emodin alone or in combination with cytarabine (Ara-C) on multidrug-resistant AML HL-60/ADR cells and in a mouse xenograft model of human highly tumorigenic AML HL-60/H3 cells. The underlying mechanism was also addressed. METHODS: Cell viability after treatment was measured by MTT assay. The DNA fragmentation assay, Annexin V-PE/7-AAD, AO/EB staining, and electron microscopy were introduced to assess the apoptotic induction effects. Changes in protein expression in the Akt and ERK signaling pathways were determined by western blotting. In vivo antileukemia effects on HL-60/H3 xenograft model and overall mouse survival outcomes were further analyzed in this study. RESULTS: Emodin dose-dependently induced growth inhibition and apoptotic effects in resistant HL-60/ADR cells in vitro as well as in the HL-60/H3 xenograft models in vivo. Moreover, emodin significantly enhanced chemosensitivity of AML cells to Ara-C, inhibited leukemic cell growth, and improved survival in the mouse xenograft model of AML. Dual targeting of Akt and ERK signaling pathways might contribute to the anti-leukemia effects on AML cells in vitro and in vivo. CONCLUSION: Emodin and its combination with Ara-C may be considered a promising therapeutic approach in AML and worthy of further investigation.

MicroRNAs as a Novel Class of Diagnostic Biomarkers in Detection of Oral Carcinoma: a Meta-Analysis Study.

BACKGROUND: Recent studies have highlighted the potential diagnostic values of microRNAs (miRNA) in various cancers involving oral cancer. This meta-analysis sought to summarize the global diagnostic accuracy of miRNAs for patients with oral cancer (OC). METHODS: A systematic review of multiple databases was performed to obtain original studies fulfilling search criteria and the quality of studies was assessed by the QUADAS tool. The bivariate meta-analysis model was employed to plot the summary receiver operator characteristic (SROC) curve. Influence analysis, meta-regression, and publication bias assay were all conducted using Stata 12.0 software. The trim-fill adjustment method was used to further assess the possible effect of publication bias. RESULTS: A total of 8 studies were included. The SROC analysis showed that miRNA profiling allowed for the discrimination between patients with high-risk oral lesions (OC or pre-cancer) and healthy donors, with a sensitivity of 0.84 (95% CI: 0.78-0.88) and specificity of 0.83 (95% CI: 0.78-0.87), corresponding to an area under curve (AUC) of 0.90. Our subgroup analyses suggested that miRNA signature harbored higher accuracy in diagnosing oral squamous cell carcinoma (OSCC) than pre-cancer lesions (AUC, sensitivity, and specificity of 0.90, 0.83, or 0.82, respectively). Moreover, stratified analyses revealed that parallel miRNA profiling, plasma- and Caucasian-based analyses all conferred promising accuracies for OC detection. The funnel plot assay manifested evidence of a publication bias. After the adjustment by the trim and fill method, the pooled adjusted efforts were slightly attenuated. CONCLUSIONS: MiRNA profiles hallmark a potential diagnostic value for detection of OC and potentially malignant disorders. Further studies should be performed to rigorously evaluate the diagnostic accuracy of miRNA profiling for OC.

Genotype GG of rs895819 Functional Polymorphism Within miR-27a Might Increase Genetic Susceptibility to Colorectal Cancer in Han Chinese Population.

BACKGROUND: MicroRNA-27a (miR-27a) is supposed to be an oncogene in various types of cancers, and genetic variation of miR-27a might result in aberrant expression and abnormal second structure of mature-miR-27a, contributing to elevated genetic risk and poor prognosis for colorectal cancer (CRC). METHODS: In order to explore the possible association between rs895819 within miR-27a and CRC in Han Chinese population, we investigated the genotype distributions of rs895819 in 508 CRC cases and 562 healthy check-up controls using TaqMan genotype discrimination system, and analyzed the possible association between them. Odds ratio (OR) and 95% confidential interval (95% CI) were used to assess the strength between allele and genotype of the locus and risk of CRC. RESULTS: In our study, we found that genotype GG of rs895819 was significantly associated with an increased risk for CRC (17.1% vs. 11.6%, adjusted OR = 1.546, 95% CI = 1.070-2.236), and allele A carrier (AA/AG) was significantly associated with a decreased risk for CRC (82.9% vs. 89.4%, adjusted OR = 0.63, 95% CI = 0.446-0.893). In addition, a significant association was observed between genotype GG and larger tumor size (>5 cm; P < 0.001), and allele G was significantly associated with higher pathological stage (TNM-III) (P = 0.008). CONCLUSION: These results indicated that miR-27a might be involved in the development and progression of CRC, genotype GG within rs895819 might be a genetic susceptible factor for CRC. Further multicentral, large sample size, and well-designed epidemiological study as well as functional study are warrant to verify our findings.

Diagnostic value of circulating microRNAs as biomarkers for breast cancer: a meta-analysis study.

Recent studies have provided new insights into the diagnostic value of circulating microRNAs (miRNAs) for breast cancer (BCa). However, the inconsistent results between studies have prevented the widespread usage of miRNAs in clinics. To systematically assess the potential diagnostic value of circulating miRNAs in BCa, we performed a comprehensive meta-analysis. Eligible studies were retrieved by searching electronic databases. The quality of the studies was assessed on the basis of quality assessment for studies of diagnostic accuracy (QUADAS) criteria. The bivariate meta-analysis model was employed to summarize the diagnostic indices and plot the summary receiver operator characteristic (SROC) curve. A total of 15 studies were included in this meta-analysis, involving 1368 BCa patients and 849 healthy controls. Our bivariate random effects meta-analysis yielded an area under curve (AUC) value of 0.9217, with a sensitivity of 0.82 (95 % confidence interval (CI) 0.80-0.83) and specificity of 0.82 (95% CI 0.80-0.85) for the use of miRNAs in differentiating BCa patients from healthy controls. Notably, our subgroup analysis suggested that a combination of multiple miRNAs (AUC, sensitivity, and specificity of 0.9518, 0.87, and 0.88, respectively) seemed to harbor higher accuracy than single miRNA-based assays (AUC, sensitivity, and specificity of 0.8923, 0.79, and 0.77, respectively). Altogether, our data indicate that circulating miRNA profiling has a potential to be used as a screening test for BCa, among which, the detection of a combined multiple miRNAs may be a more comprehensive indicator than individual miRNA.

Diagnostic Accuracies of the TUNEL, SCD, and Comet Based Sperm DNA Fragmentation Assays for Male Infertility: a Meta-analysis Study.

BACKGROUND: Recent studies have provided new insights into the diagnostic value of sperm DNA fragmentation (SDF) for male factor sterility. This study aimed to systematically evaluate the diagnostic accuracy of the SDF test for male infertility. METHODS: Eligible studies were retrieved by searching electronic databases. The quality of the studies was assessed on the basis of quality assessment for studies of diagnostic accuracy (QUADAS) criteria tool. The bivariate metaanalysis model was employed to summarize the diagnostic indices and plot the summary receiver operator characteristic (SROC) curve by using Meta-disc 1.4 software. Influence analysis, meta-regression, and publication bias assay were all conducted through Stata 12.0 software. RESULTS: Our bivariate random effect meta-analysis yielded an AUC (area under curve) value of 0.9211 with a sensitivity (95% confidence interval) of 0.80 (0.78 - 0.82) and specificity of 0.83 (0.80 - 0.86) for the use of the SDF test in differentiating infertile males from normal fertile controls. Moreover, our subgroup analysis suggested that SDF analysis with a single TUNEL test resulted in an AUC value of 0.9506, with a pooled sensitivity of 0.77 (0.74 - 0.80) and specificity of 0.91 (0.87 - 0.94), while SCD and Comet assays displayed a combined sensitivity of 0.77 (0.67 - 0.81) or 0.91 (0.88 - 0.94), and specificity of 0.84 (0.75 - 0.91) or 0.63 (0.54 - 0.70), accompanied by an AUC value of 0.8408 or 0.9473. CONCLUSIONS: The SDF assay confers a relatively high diagnostic accuracy for infertility detection, among which the TUNEL based methodology seems to achieve higher accuracy than the SCD and Comet assays.

Thrombolytic effects in vivo of nattokinase in a carrageenan-induced rat model of thrombosis.

BACKGROUND/AIMS: Nattokinase is a serine protease produced by Bacillus subtilis during the fermentation of the soybean product natto. The fibrinolytic activity and thrombolytic effects of nattokinase have been observed in vitro, but the effect in vivo has still to be researched. The objective of this study was to demonstrate the activity of nattokinase in vivo. METHODS: To establish a rat model of thrombosis, κ-carrageenan was injected subcutaneously into the toes of Sprague-Dawley (SD) rats. Histological examination confirmed thrombosis. The rats were then treated with varying doses of nattokinase and the resulting thrombolysis was histologically assessed. ELISA was used to determine the levels of the fibrin/fibrinogen degradation products (FDPs) and D-dimer, which are sensitive indices of fibrinolytic activity. Vermis kinase, a known thrombolytic agent, was used as a positive control. RESULTS: Biopsy results revealed partial thrombolysis in the tail vessels of the rats treated with nattokinase or vermis kinase. FDP and D-dimer levels were higher in rats treated with high-dose nattokinase than in those treated with saline. No difference in FDP or D-dimer levels was observed between rats treated with high-dose nattokinase and those treated with vermis kinase. CONCLUSIONS: Both the histological and physiological evidence from this study indicate that nattokinase exerts thrombolytic effects in vivo.

[Effect of Sunitinib on Pyroptosis of Drug-Resistant Leukemia K562/ADR Cells and Its Pathway].

OBJECTIVE: To investigate the inducing effect of sunitinib on the death of drug-resistant leukemia K562/ADR cells and the related signaling pathway. METHODS: K562/ADR cells were treated with different concentrations of sunitinib, and the cells were collected at 24, 48, 72, and 96 hours, respectively. MTS assay was used to detect the effect of sunitinib on the proliferation of K562/ADR cells, and the appropriate sunitinib intervention time and concentration were determined. QPCR and Western blot were used to detect the mRNA and protein expression levels of apoptosis-related genes in K562/ADR cells treated with sunitinib. Four different cell death inhibitors Nec-1, VX-765, CQ and Fer-1 were used to detect the death mode of K562/ADR cells treated with sunitinib. QPCR and Western blot were used to detect the mRNA and protein expression levels of pyroptosis-related genes in K562/ADR cells treated with sunitinib. RESULTS: Sunitinib significantly inhibited the proliferation of K562/ADR cells in a time - and concentration-dependent manner(R48 H=0.9579, r4 µg/ml=0.9740). The IC50 of sunitinib was (3.96±0.14) µg/ml at 48 hours. The mRNA and protein expression levels of apoptosis-related genes Bax, BCL-2 , Caspase-3 and Caspase-9 in K562/ADR cells treated with sunitinib did not change significantly. After treatment with four different cell death inhibitors, only the pyroptosis inhibitor VX-765 could significantly reverse the inhibitory effect of sunitinib on the proliferation of K562/ADR cells (P<0.01). The mRNA and protein expression levels of pyroptosis-related genes Caspase-1, Caspase-4, Caspase-5, NLRP3, GSDMD and IL-1ß in K562/ADR cells treated with sunitinib were significantly increased (P<0.01). CONCLUSION: Sunitinib can induce pyroptosis in drug-resistant leukemia K562/ADR cells. Further study of the signaling pathways related to pyroptosis may provide experimental basis for the treatment of drug-resistant leukemia.

Comprehensive analysis of ferroptosis-related gene signatures as a potential therapeutic target for acute myeloid leukemia: A bioinformatics analysis and experimental verification.

Background: Ferroptosis plays an important role in the development of acute myeloid leukemia (AML); however, the exact role of ferroptosis-related genes in the prognosis of AML patients is unclear. Methods: RNA sequencing data and the clinicopathological characteristics of AML patients were obtained from The Cancer Genome Atlas database, and ferroptosis-related genes were obtained from the FerrDb database. Cox regression analysis and least absolute shrinkage and selection operator analysis were performed to identify ferroptosis-related gene signatures. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and single-sample gene set enrichment analysis (ssGSEA) were performed to explore the biological functions of the ferroptosis-related genes. Finally, ferroptosis of AML cells was induced by erastin and sulfasalazine to detect the changes in the expression of relevant prognostic genes and explore the underlying mechanisms using quantitative real-time polymerase chain reaction (qRT-PCR). Results: Seven ferroptosis-related gene signatures (SOCS1, ACSF2, MYB, EIF2AK4, AIFM2, SLC7A11, and GPX4) were identified in the training group. Kaplan-Meier and Cox regression analyses confirmed that risk score was an independent prognostic predictor of AML in the training and validation groups (P<0.05). Further, functional enrichment analysis revealed that seven ferroptosis-related genes were associated with many immune-related biological processes. Most importantly, erastin and sulfasalazine can induce the ferroptosis of AML cells. Overall, SLC7A11 and the SLC7A11/xCT-GSH-GPX4 pathway may be the respective key gene and potential regulatory pathway in erastin- and sulfasalazine-induced ferroptosis of AML cells. Conclusions: A novel signature involving seven ferroptosis-related genes that could accurately predict AML prognosis was identified. Further, the Food and Drug Administration-approved drug, sulfasalazine, was demonstrated for the first time to induce the ferroptosis of AML cells. SLC7A11 and the SLC7A11/xCT-GSH-GPX4 pathway may be the respective key gene and underlying mechanism in this process, ultimately providing new insights into the strategies for the development of new AML therapies.