Interleukin 35 contributes to immunosuppression by regulating inflammatory cytokines and T cell populations in the acute phase of sepsis.

Cytokines interact closely with each other and play a crucial role in the progression of sepsis. We focused on the associations of a cytokine network with IL-35 in sepsis. First, the retrospective study included 42 patients with sepsis and 23 healthy controls. Blood samples were collected from patients on days 1, 2, 4. Levels of IL-35, IL-1ß, IL-4, IL-6, IL-10, IL-17A, TNF-α and IFN-γ were measured. They all increased to various extend on days 1, 2, 4, and strongly associated with markers of disease severity. Network analysis revealed a network formed by IL-35, with IL-6, IL-10, IL-17A, TNF-α and IFN-γ throughout the acute phase of sepsis(days 1, 2 and4). Then, the CLP-induced septic rats were used. The recombinant human IL-35(rIL-35) upregulated the levels of IL-10, but downregulated IL-4, IL-6, IL-17A, TNF-α and IFN-γ, while it had no significant effect on IL-1ß, and upregulated the percentages of CD4+CD25+Tregs, and iTR35, but downregulated Teff cells in the peripheral blood. The rIL-35 reduced inflammation damage and improved prognosis of the septic rats. IL-35 forms a network with other cytokines and plays a major role in the immunopathogenesis of sepsis.

Expression profiling of N6-methyladenosine modified circRNAs in acute myeloid leukemia.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults, associated with poor prognosis and easy relapse of disease. Circular RNAs (circRNAs) were detected to be m6A modified and the role of m6A circRNAs has been reported in other diseases including cancers, however, their role has not been elucidated in AML yet. In the present study, we aimed to investigate the expression profiling of m6A circRNAs in AML. We performed m6A circRNAs microarray analysis to identify differentially expressed m6A circRNAs in bone marrow samples from AML patients and healthy individuals (control). Furthermore, bioinformatics analysis predicted the potential functions and relevant pathways that may be associated with the m6A circRNAs. The circRNA m6A methylation levels were found to be positively associated with the circRNAs expression, suggesting circRNA m6A modification could contribute to circRNA regulation in AML. Further analysis demonstrated that circRNA m6A modification might influence the circRNA-miRNA-mRNA co-expression network that may contribute to the circRNA regulatory network in AML. Our findings provide evidence of the differential expression profile of m6A circRNAs in AML, and circRNA m6A modification may contribute to circRNA regulatory function in AML.

Effects of Continuous Blood Purification without Heparin on Strem-1, NSE, and IL-10 Levels In Patients With Sepsis.

One of the most common causes of mortality and disability across the world is sepsis, a condition marked by an abnormal immunological reaction in the body. The prognosis of patients with sepsis is much improved when they are diagnosed early and provided with proper therapy. Soluble triggering receptors expressed on myeloid cells (STREM-1), Interleukin-10 (IL-10), neuronal serum enolase (NSE), and so on are possible biomarkers for the diagnosis of sepsis based on the pathophysiology. Blood purification treatment to control the cytokine storm induced by sepsis was regarded to be promising. Recently, the treatment of sepsis is likely to shift toward a multimodal approach. Hence in this paper, we investigated the effect of a continuous blood purification technique (hemofiltration without heparin followed by hemoadsorption) on STREM-1, NSE, and IL-10 levels in patients suffering from sepsis. A sample of hundred patients suffering from sepsis was randomly allocated to one of the two groups (study and control groups). The control group got standard sepsis care, whereas the research group got continuous blood purification treatment. To compare the differences between the two groups, we used t-statistical analysis. Blood STREM-1, IL-10, and NSE concentrations of the study group were significantly lesser than that of the control group after therapy. As a result, continuous blood purification can significantly minimize the dysregulated immune response in patients with sepsis, promote immune function and improve the survival rate.

miR-34b-5p promotes renal cell inflammation and apoptosis by inhibiting aquaporin-2 in sepsis-induced acute kidney injury.

OBJECTIVE: This study was designed to uncover the mechanism of miR-34b-5p-mediated aquaporin-2 (AQP2) in sepsis-induced injury using human renal tubular epithelial cells (HK-2). METHODS: Serum levels of miR-34b-5p, TNF-α, IL-1ß, IL-6, serum creatinine (SCr), and blood urea nitrogen (BUN) in septic patients with acute kidney injury (AKI) and healthy controls were detected. Lipopolysaccharide (LPS) was used to induce sepsis in HK-2 cells. LPS-induced HK-2 cells were transfected with miR-34b-5p inhibitor, miR-34b-5p mimic, pcDNA3.1-AQP2, si-AQP2, miR-34b-5p inhibitor + si-NC, or miR-34b-5p inhibitor + si-AQP2. The expressions of miR-34b-5p, AQP2, Bax, Bcl-2, cleaved caspase-3, TNF-α, IL-1ß, and IL-6 in HK-2 cells were detected. TUNEL staining revealed the apoptosis of HK-2 cells. Dual-luciferase reporter assay verified the binding between miR-34b-5p and AQP2. RESULTS: The expression of miR-34b-5p and the inflammatory responses were augmented in septic AKI patients. miR-34b-5p was up-regulated and AQP2 was down-regulated in LPS-induced HK-2 cells. miR-34b-5p inhibition or AQP2 overexpression ameliorated apoptosis and inflammation in LPS-induced HK-2 cells. In contrast, overexpressing miR-34b-5p deteriorated LPS-induced injury in HK-2 cells. AQP2 was a downstream target of miR-34b-5p. AQP2 silencing abolished the suppressive effects of miR-34b-5p inhibition on LPS-induced apoptosis and inflammatory response in HK-2 cells. CONCLUSION: miR-34b-5p inhibits AQP2 to promote LPS-induced injury in HK-2 cells.

Effects of Novel ncRNA Molecules, p15-piRNAs, on the Methylation of DNA and Histone H3 of the CDKN2B Promoter Region in U937 Cells.

Non-coding RNAs (ncRNAs) play key roles in epigenetic events. However, the exact mechanism of ncRNA guidance, particularly piwi-interacting RNAs (piRNAs), for the targeting of epigenetic regulatory factors to specific gene regions is unclear. Although piRNA function was first established in germ-line cells, piRNA may be crucial in cancer cells. This study investigated the potential roles of CDKN2B-related piRNA in leukemia cells to provide a potential tumorigenesis model of leukemia. CDKN2B-related piRNAs, hsa_piR_014637 and hsa_piR_011186 were transduced into the leukemia cell line U937 to study the effect of these two piRNAs on cell-cycle progression, apoptosis, heterochromatin formation, CDKN2B methylation and expression. Our results show that over-expressing hsa_piR_011186 promoted cell-cycle progression and decreased apoptosis. We also observed inhibition of CDKN2B gene expression. These effects were likely mediated by novel piRC (piRNA complex) of CDKN2B-related piRNA that associate with DNMT1, Suv39H1 and/or EZH2 proteins to modulate the methylation of DNA and histone H3 in the promoter region of the CDKN2B gene. The novel piRC complex facilitated epigenetic modifications on the promoter of cell-cycle regulating genes, providing an expanded view of the role of piRNA in the progression of leukemia cells.

Growth Inhibition Accompanied by MOB1 Upregulation in Human Acute Lymphoid Leukemia Cells by 3-Deazaneplanocin A.

Our purpose was to investigate the effect of 3-deazaneplanocin A (DZNep) on human T-cell acute lymphoid leukemia (T-ALL) cells, and to explore the underlying molecular mechanisms. The human T-ALL cell line Molt4 was treated with DZNep, and cell proliferation was examined. The expression of Mps one binder kinase activator 1 gene (MOB1) mRNA and protein was determined by RT-PCR and Western blotting, respectively. The histone modification effect of DZNep on the lysine 9 of histone 3 associated with MOB1 promoters was examined with chromatin immunoprecipitation and quantitative PCR, and CpG methylation in MOB1 promoters was detected by bisulfite sequencing PCR. DZNep treatment inhibited the growth of Molt4 cells. The expressions of MOB1 genes were upregulated by DZNep treatment, and histone methylations in their promoters were significantly reduced. The results indicate that DZNep is a promising therapeutic compound for the treatment of human T-ALL.

[Hypermethylation of CpG island of DLC-1 gene and arsenic trioxide-induced DLC-1 gene demethylation in multiple myeloma].

OBJECTIVE: To explore the role of hypemethylation of DLC-1 gene in the pathogenesis of multiple myeloma (MM) and examine the effects of arsenic trioxide (As(2)O(3))-induced demethylation of DLC-1 gene in U266 cell line. METHODS: The methylation status of DLC-1 gene was detected by methylation specific PCR (MSP) in MM patients from 2008 to 2012. And the expression of DLC-1 gene mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). Methylation statuses of DLC-1 gene exposed to As(2)O(3) were detected by bisulfite sequencing PCR (BSP). And the mRNA expressions of DLC-1 and DNA methyltransferase (DNMT1, T3a and 3b) were determined by real-time fluorescence quantitative PCR (RTFQ-PCR). RESULTS: Hypermethylation of CpG island of DLC-1 gene was observed in 37/52 (71.15%) MM patients. DLC-1 gene was not expressed after methylation. As(2)O(3) could induce DLC-1 gene demethylation. After 72-houe treatments of 0.5, 1.0 and 2.0 µmol/L As(2)O(3), the methylation rate of DLC-1 gene dropped from 95.38% to 63.07%, 30.00% and 7.69%. As compared with the untreated group, the expression of DLC-1 gene mRNA increased to (1.60 ± 0.09), (3.66 ± 0.17) and (5.29 ± 0.15) folds after exposures (all P < 0.05) . And As(2)O(3) could induce the expression of DNMT1, DNMT3a, DNMT3b gene mRNA (all P < 0.05). CONCLUSIONS: Methylation of DLC-1 gene is essential in the pathogenesis of MM and may provide a new diagnostic technique and drug target for the treatment of MM. And As(2)O(3) may activate the expression of DLC-1 gene through demethylation.

NLRP3 inflammasome inhibitor MCC950 can reduce the damage of pancreatic and intestinal barrier function in mice with acute pancreatitis.

PURPOSE: Abnormal activation of NOD-like receptor protein 3 (NLRP3) inflammasome can lead to the occurrence and progression of acute pancreatitis. This study investigated the protective effect of MCC950 on pancreatitis mice. METHODS: Eighteen mice were randomly divided into control group, severe acute pancreatitis (SAP) group and SAP+MCC950 group. Serum interleukin (IL)-1ß, IL-6 and tumor necrosis factor-α (TNF-α) were measured by ELISA. Hematoxylin and eosin (HE) staining was used to evaluate the pathological damage. Western blotting was used to detect the expression of NLRP3 inflammasome and tight junction proteins in the small intestine and pancreas. RESULTS: MCC950 could reduce the levels of IL-6 and IL-1ß in SAP mice. After treatment with MCC950, the expression levels of NLRP3 inflammasome in the pancreas of SAP mice were significantly reduced and the pathological damage to the pancreas and intestine was alleviated. Compared with the control group, the expression of tight junction protein (ZO-1,occludin and claudin-4) in the intestinal mucosa of SAP mice was decreased, and the expression of claudin-4 and occludin were upregulated after MCC950 treatment. CONCLUSIONS: MCC950 can inhibit NLRP3 inflammasome activation and significantly reduce the inflammatory response and delay the process of pancreatitis. It has therapeutic potential in the treatment of acute pancreatitis.

Biomarkers for the early prediction of contrast-induced nephropathy after percutaneous coronary intervention in adults: A systematic review and meta-analysis.

Contrast-induced nephropathy (CIN) is a complication of patients undergoing percutaneous coronary intervention (PCI). Promising biomarkers for the early prediction of CIN can significantly improve outcomes of these patients. We searched PubMed, EMBASE, Web of Science, and Cochrane Library for studies. Trials reporting an area under the curve (AUC) for the utility of novel biomarkers in the early prediction of CIN in adults after PCI were included. In total, 42 studies comprising 11,984 adult patients undergoing PCI met the criteria. Four urinary biomarkers and four blood biomarkers were included. For urine biomarkers, the pooled AUCs for neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), liver-type fatty acid-binding protein (L-FABP), and kidney injury molecule-1 (KIM-1) were 0.91 (95% CI 0.89-0.94), 0.79 (0.75-0.82), 0.78 (0.74-0.82), and 0.79 (0.76-0.83), respectively. The blood biomarkers NGAL, cystatin C, brain natriuretic peptide (BNP), and C-reactive protein (CRP) had pooled AUCs of 0.93 (0.91-0.95), 0.92 (0.89-0.94), 0.78 (0.74-0.81), and 0.75 (0.71-0.79), respectively. Subgroup analysis showed that blood NGAL in early CIN predictive time (<6 h) was more effective in predicting CIN. The efficiency of cystatin C in predicting CIN was reduced, whereas that of L-FABP was increased among chronic kidney disease (CKD) patients.

Epigenetic modifications in acute myeloid leukemia: The emerging role of circular RNAs (Review).

Canonical epigenetic modifications, which include histone modification, chromatin remodeling and DNA methylation, play key roles in numerous cellular processes. Epigenetics underlies how cells that posses DNA with similar sequences develop into different cell types with different functions in an organism. Earlier epigenetic research has primarily been focused at the chromatin level. However, the number of studies on epigenetic modifications of RNA, such as N1­methyladenosine, 2'­O­ribosemethylation, inosine, 5­methylcytidine, N6­methyladenosine (m6A) and pseudouridine, has seen an increase. Circular RNAs (circRNAs), a type of RNA species that lacks a 5' cap or 3' poly(A) tail, are abundantly expressed in acute myeloid leukemia (AML) and may regulate disease progression. circRNAs possess various functions, including microRNA sponging, gene transcription regulation and RNA­binding protein interaction. Furthermore, circRNAs are m6A methylated in other types of cancer, such as colorectal and hypopharyngeal squamous cell cancers. Therefore, the critical roles of circRNA epigenetic modifications, particularly m6A, and their possible involvement in AML are discussed in the present review. Epigenetic modification of circRNAs may become a diagnostic and therapeutic target for AML in the future.

[The establishment and evaluation of diagnostic accuracy of AllGlo(TM) probe-based techniques for invasive aspergillosis.].

OBJECTIVE: To establish and evaluate a diagnostic technique based on the AllGlo(TM) probe for the invasive aspergillosis. METHODS: With the self-designed AllGlo(TM) probes and primers and the standards, two standard curves of the real-time PCR based on AllGlo(TM) probes were established respectively for A. flavus and A. fumigatus, then its specificity, sensitivity and reproducibility were evaluated respectively. RESULTS: The findings indicated that the standard curve of A. flavus was Y = -3.003X + 36.825, and A.fumigatus' was Y = -3.052X + 38.016, and their interassay coefficient of variation respectively were 15.60% and 12.94%, suggesting a good reproducibility. The lowest spore concentration they could be detected was 10 CFU/ml, which equated to 100 - 1000 copies of internal transcribed spacer (ITS)2 genes, suggesting a good sensibility. They didn't have cross-positive reaction with other fungus, human genome and bacteria, which indicated a good specificity. CONCLUSION: The diagnostic technique based on the AllGlo(TM) probe for the invasive aspergillosis possessed a good sensitivity, good specificity and deadly accuracy.

[Hsa-let-7b-5p Inhibits Proliferation of Human Leukemia THP-1 Cells via FTO/m6A/MYC Signaling Pathway].

OBJECTIVE: To investigate the down-regulation effect of let-7b-5p on the expression of FTO in acute myeloid leukemia cell line THP-1 and inhibitory effect on THP-1 proliferation via m6A/MYC signaling pathway. METHODS: The acute myeloid leukemia cell line THP-1 and the normal human peripheral blood mononuclear cells (PBMNC) were selected as subjects. The expression of let-7b-5p and FTO mRNA in those cells was detected by qPCR, further the expression of FTO protein in those cells was detected by Western blot. And, the luciferase reporter gene assay was used to verify the targeting effect of let-7b-5p on FTO. Finally, THP-1 cells were transfected respectively with let-7b-5p mimic, and PBMNC with let-7b-5p inhibitor, there after the C-MYC mRNA m6A enrichment level in transfected cells was analyzed by dot blot, the expression levels of let-7b-5p, FTO and c-MYC were assayed by RT-PCR, the expressions of FTO and c-MYC protein were verified by Western blot, and the proliferation level of cells after transfection was detected by MTT assay. RESULTS: Compared with PBMNC, the expression of let-7b-5p in THP-1 significantly decreased, while the expression of FTO was significantly increased (P<0.05). After transfection with let-7b-5p mimic combined with FTO 3'-UTR, the luciferase activity of transfected THP-1 cells significantly decreased, but the luciferase activity significantly increased after transfection with mutant 3'-UTR, which was significantly different from the negative control group(blank vector) (P<0.05). Let-7b-5p inhibitor down-regulated c-MYC mRNA m6A enrichment, and then up-regulated the expression of FTO in transfected PBMNC cells, the effects of which were significant (P<0.05). However, let-7b-5p mimic up-regulated c-MYC mRNA m6A enrichment level and down-regulated the expression of FTO in the transfected THP-1 cells, and the cell proliferation rate was significantly lower than that in the negative control group (blank vector) (P<0.05). CONCLUSION: Human acute myeloid leukemia cell line THP-1 low expresses the let-7b-5p, which regulates c-MYC expression through let-7b-5p-/FTO-/m6A axis and promotes the proliferation of leukemia cell line THP-1.

NLRP3 inflammasome inhibitor MCC950 can reduce the damage of pancreatic and intestinal barrier function in mice with acute pancreatitis

Purpose: Abnormal activation of NOD-like receptor protein 3 (NLRP3) inflammasome can lead to the occurrence and progression of acute pancreatitis. This study investigated the protective effect of MCC950 on pancreatitis mice. Methods: Eighteen mice were randomly divided into control group, severe acute pancreatitis (SAP) group and SAP+MCC950 group. Serum interleukin (IL)-1ß, IL-6 and tumor necrosis factor-α (TNF-α) were measured by ELISA. Hematoxylin and eosin (HE) staining was used to evaluate the pathological damage. Western blotting was used to detect the expression of NLRP3 inflammasome and tight junction proteins in the small intestine and pancreas. Results: MCC950 could reduce the levels of IL-6 and IL-1ß in SAP mice. After treatment with MCC950, the expression levels of NLRP3 inflammasome in the pancreas of SAP mice were significantly reduced and the pathological damage to the pancreas and intestine was alleviated. Compared with the control group, the expression of tight junction protein (ZO-1,occludin and claudin-4) in the intestinal mucosa of SAP mice was decreased, and the expression of claudin-4 and occludin were upregulated after MCC950 treatment. Conclusions: MCC950 can inhibit NLRP3 inflammasome activation and significantly reduce the inflammatory response and delay the process of pancreatitis. It has therapeutic potential in the treatment of acute pancreatitis.

Relationship between epigenetic changes in Wnt antagonists and acute leukemia.

The present study was designed to investigate the relationship among epigenetic changes in Wnt antagonists, histone H4K20me1 and the expression of tumor-suppressor genes in acute leukemia (AL) to better understand the pathogenesis of leukemia. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect messenger RNA (mRNA) expression levels of Wnt antagonists (Wnt5a, HDPR1, DKK1 and DKK3) in patients with AL and in normal controls; pyrophosphate sequencing was performed to detect the methylation status of the Wnt5a promoter; and western blotting was performed to detect the overall expression levels of Wnt5a protein and histone H4K20me1 in patients with acute myeloid leukemia (AML) and in normal controls. The relationship between Wnt5a protein expression and histone H4K20me1 was analyzed. Chromatin immunoprecipitation-qPCR (ChIP-qPCR) was performed to investigate the recruitment of H4K20me1 and SET8 to the Wnt5a promoter and coding regions. Our results demonstrated that the expression levels of Wnt antagonists were generally low in AML, but showed differential expression in acute lymphocytic leukemia (ALL). In most cases of AML, methylation of the Wnt5a promoter was observed and Wnt5a protein expression was low. In some cases of AML, the overall level of H4K20me1 protein was higher than that in normal controls. In addition, Wnt5a expression was positively correlated with H4K20me1 expression and was unrelated to the methylation status of its promoter. Moreover, H4K20me1 and SET8 were enriched in the Wnt5a promoter region and coding region. By contrast, Wnt5a expression was unrelated to H4K20me1 expression in normal controls. Moreover, we observed that the methylation of Wnt antagonists was often found in patients with AL, particularly those with AML, whereas the extent of methylation was variable in ALL patients. Wnt5a expression was positively correlated with the enrichment of H4K20me1 and SET8 at the Wnt5a promoter and coding regions. H4K20me1 increased Wnt5a expression by promoting transcription initiation and elongation.

CpG island methylator phenotype and its relationship with prognosis in adult acute leukemia patients.

OBJECTIVE: To investigated the relationship between CpG island methylator phenotype (CIMP) and prognosis in adults with acute leukemia. METHODS: Bone marrow samples from 53 acute myeloid leukemia and 50 acute lymphoblastic leukemia patients were collected. The methylation status of 18 tumor suppressor genes was determined using methylation-specific polymerase chain reaction. RESULTS: Greater than 30% of acute leukemia patients had methylated p15, p16, CDH1, CDH13, RUNX3, sFRP1, ID4, and DLC-1 genes; methylation of ≥4 were defined as CIMP positive. Age, type of leukemia, white blood cell count, and CIMP status were significantly associated with recurrence-free survival (RFS) and overall survival (OS) (P < 0.05). CIMP status was an independent prognostic factor for OS (hazard ratio: 2.07, 95% confidence interval: 1.03-4.15, P = 0.040). CIMP-negative patients had significantly improved RFS and OS (P < 0.05). p16 and DLC1 methylation was significantly associated with RFS and OS (P < 0.05). CONCLUSIONS: CIMP may serve as an independent risk factor for evaluating the prognosis of patients with acute leukemia.

Overexpression of microRNA-143 inhibits growth and induces apoptosis in human leukemia cells.

Microrna-143 (miR-143) has been suggested to be a tumor suppressor, yet its role in hematological tumors has not been determined. Thus, we aimed to explore the expression and function of miR-143 in leukemia cells. miR-143 expression was assessed in bone marrow samples from 63 leukemia patients and 15 healthy controls using q-PCR, and its correlation with DNMT3A expression was determined. In addition, after lentiviral-mediated miR-143 overexpression, K562 cell proliferation was evaluated using CCK-8 analysis; cell cycle progression and apoptosis were determined using flow cytometry. The expression of Bcl-2 and pro-caspase-3 and -9 was assessed by q-PCR and western blot analysis, respectively. Leukemia patients had significantly lower relative miR-143 expression than healthy controls (P=0.004), and the expression levels of miR143 and DNMTA3A were negatively correlated (r=-0.663, P=0.001). Overexpression of miR-143 decreased DNMT3A mRNA and protein expression, and significantly reduced K562 cell proliferation at 72 and 96 h (both P ≤ 0.018). In addition, reduced colony formation and cell cycle progression were observed upon miR-143 overexpression. Flow cytometric analysis revealed that the early apoptosis rate was higher in the miR-143 group than the rate in the NC group. Bcl-2 mRNA expression and pro-caspase-3 and -9 protein expression were reduced in the miR-143-expressing cells. These findings suggest that miR-143 plays an important role in leukemia cell proliferation and apoptosis, possibly through silencing of DNMT3A. Further studies are necessary to determine the prognostic value and therapeutic potential of targeting miR-143.

Epigallocatechin-3-gallate and trichostatin A synergistically inhibit human lymphoma cell proliferation through epigenetic modification of p16INK4a.

DNA methylation and histone deacetylation play important roles in the occurrence and development of cancers by inactivating the expression of tumor suppressors, including p16(INK4a), a cyclin-dependent kinase inhibitor. The present study investigated the effect of epigallocatechin-3-gallate (EGCG) alone or in combination with trichostatin A (TSA) on p16(INK4a) gene expression and growth in human malignant lymphoma CA46 cells. CA46 cell viability and cell cycle were analyzed; methylation of the p16(INK4a) gene was assessed by nested methylation-specific PCR (n-MSP). p16(INK4a )mRNA and protein expression was determined by real-time quantitative PCR and western blot analyses, respectively. Both EGCG and TSA alone inhibited CA46 cell proliferation; the combined treatment (6 µg/ml EGCG and 15 ng/ml TSA) significantly reduced CA46 cell proliferation from 24 to 96 h (all P<0.001). Cells treated with 24 µg/ml EGCG or the combination treatment (6 µg/ml EGCG and 15 ng/ml TSA) had lower proliferative indices when compared to the other groups. Co-treatment with EGCG and TSA decreased p16(INK4a) gene methylation, which coincided with increased p16(INK4a) mRNA and protein expression. Thus, EGCG and TSA synergistically reactivate p16(INK4a) gene expression in part through reducing promoter methylation, which may decrease CA46 cell proliferation.

[Expression of Hippo signaling pathway core element MST1 in acute leukemia patients and its significance].

This study was aimed to investigate the expression of Hippo signaling pathway core element MST1 gene in acute leukemia (AL) patients, and explore its relation with AL pathogenesis and prognosis. 50 newly diagnosed patients with AL, 33 normal people, 23 patients with AL in complete remission, 12 refractory or relapsed patients were tested for the expression of MST1 gene by real-time quantitative PCR, Western blot was used to further validate the level of MST1 protein expression. And combined with clinical data, prognostic factors of the patients were analyzed. The results indicated that compared with the normal people, the expression level of MST1 in newly diagnosed patients with AL significantly decreased (P < 0.05), but significantly increased in AL patients with complete remission (CR), the difference of expression was statistically significant before CR and after CR (P < 0.05). Compared with refractory or relapsed patients, the expression level of MST1 gene in newly diagnosed patients was not significantly different (P > 0.05). Besides, the expression level of MST1 between the patients with CR and the normal people was not significantly different (P > 0.05). It is concluded that the low expression of MST1 may be related with the pathogenesis and prognosis of AL.

[Differential expression of miR-143 in bone marrow cells between acute leukemia patients and normal people and its significance].

MicroRNA abnormality is closely related to the development of acute leukemia. This study was aimed to investigate the differential expression of miR-143 in bone marrow cells between patients with acute leukemia and normal people. Bone marrow cells from 50 AL patients and 20 normal people were collected respectively and the total RNA was isolated routinely. The quantitative real-time PCR (Q-PCR) method was used to detect the expression levels of miR-143 in these specimens. The results showed that compared with the normal people, the expression of miR-143 in AL patients was significantly decreased (p < 0.05), which significantly increased after complete remission; besides, the expression of miR-143 was negatively correlated with the expression of DNMT3A mRNA, a known target gene of miR-143. It is concluded that the expression of miR-143 in bone marrow cells of AL patients is down-regulated which may be related with the development of acute leukemia.

[Methylation status of IEX-1 gene promotor CpG island in malignant hematopoietic cell lines].

This study was to purposed to explore the methylation status changes of IEX-1 gene promoter CpG island and its relevance with occurrence of hematologic malignancies. The methylation status of IEX-1 gene promoter CpG island in 9 NB4, HL-60 U937, Raji, CA46, Jurkat, K562, CEM and Molt4 hematologic malignant cell lines was detected by using methylation-specific PCR, the methylation status of IEX-1 gene promoter CpG island in normal peripheral blood mononuclear cells treated by M. sssI enzyme and the methylation status of IEX-1 gene promoter CpG island in normal peripheral blood mononuclear cells untreated were used as positive and negative controls respectively. The results showed that the hypermethylation of IEX-1 gene promoter CpG islands was detected in NB4, Molt4 and Raji cell lines, as well as in normal peripheral blood mononuclear cells treated by M. sssl enzyme; the partial methylation status was found in CA46, CEM, U937, K562, HL-60 and Jurkat cell lines; the unmethylation status was observed in untreated normal peripheral blood mononuclear cells. It is concluded that the changes of methylation status of gene IEX-1 promoter CpG island correlates with hematologic malignancies to a certain extent.