MicroRNA-16-5p exacerbates sepsis by upregulating aerobic glycolysis via SIRT3-SDHA axis.

Sepsis is a life-threatening condition, and treatment for sepsis in clinic is often not available, partially due to insufficient understanding of the pathogenesis of sepsis. Extensive study to elucidate the pathogenesis is required to improve the clinical management and outcome of sepsis. In this study, we investigated the pathogenesis of sepsis using peripheral blood mononuclear cells (PBMCs) from septic patients and studied the underlying mechanism of miR-16-5p on aerobic glycolysis in lipopolysaccharide (LPS)-treated THP1 and Raw264.7 cells. The levels of RNA and protein were determined by real-time quantitative PCR and immunoblotting assay, respectively. The production of high mobility group box 1 (HMGB1) was measured by enzyme-linked immunosorbent assay. The levels of succinate and lactate were determined using colorimetric kits. The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured by extracellular flux analyzer. The results showed that the expression of miR-16-5p was elevated, while sirtuin 3 (SIRT3) was decreased in PBMCs from septic patients and LPS-treated cells, along with accumulation of acetylated succinate dehydrogenase complex, subunit A. Concomitantly, an increase in HMGB1, succinate, lactate, as well as ECAR and a decrease in OCR were observed. Knockdown of miR-16-5p upregulated SIRT3 expression, facilitated SDHA deacetylation, and attenuated sepsis-related aerobic glycolysis. Further study identified that SIRT3 is targeted by miR-16-5p, and overexpression of SIRT3 rescued LPS-induced responses via deacetylation of SDHA. Our findings revealed a novel miR-16-5p-regulated SIRT3-SDHA axis in sepsis and provided novel insights for sepsis treatment.

[Analysis on prognosis and correlative factors of acute nonlymphocytic leukemia].

To analyze the prognosis and risk factors of acute nonlymphocytic leukemia (ANLL), 94 patients with acute nonlymphocytic leukemia were enrolled in this study, while survival rate and risk factors of prognosis were analyzed. The results indicated that the complete remission (CR) ratio was 51.1%. Overall survival and event-free survival rates of month 6, 12, 18, 24 were 68.6%, 55.8%, 53.8%, 46.4%, 21.3% and 57.9%, 38.6%, 33.3%, 31.6%, 0% respectively. The factors such as age<40 years, WBC<10.0x10(9)/L before chemotherapy, WBC in the period of bone marrow suppression<1.0x10(9)/L, chemotherapy within 1 month after occurrence of leukemia, blood transfusion before chemotherapy of APL had favourable influence on remission and survival rates of ANLL patients. CR1, the time to get CR, length of CR and relapse significantly correlated with prognosis (p<0.05). It is concluded that the individualized therapy concerning the risk factors should be applied to ANLL patients for improving the remission, survival rate and prognosis.

[Prognosis and related factors of acute lymphoblastic leukemia].

In order to analyze the prognosis and related factors of acute lymphoblastic leukemia (ALL), 53 newly diagnosed ALL patients were enrolled in this study. The therapeutic efficacy and prognosis of 53 cases of ALL were analyzed, the remission, relapse, overall survival and event-free survival were studied, and relation between different factors and prognosis of ALL were investigated by comparison of cases in same stage. The results showed that the complete remission was achieved in 36 out of 53 patients, the total remission rate was 67.9%, the total relapse rate was 37.7%, the median relapse duration was 6 months after remission. Median overall survival (OS) and median event-free survival (EFS) time were 4 and 1 months after remission respectively, OS and EFS rate of 18 month was 35.1% and 14.2%. The patients with different gender had significantly different EFS. Age was an independent risk factor of CR rate. White blood cell count and hemoglobin level of newly diagnosed patients were significantly correlated with OS and EFS. Absolute neutrophil count (ANC) at the end of the induction chemotherapy was an independent related factor of OS, the higher ANC, the lower risk of death. The patients with or without chemotherapy related infection had different relapse rate. The patients with bleeding after chemotherapy had lower OS when compared with those without bleeding. Serum glucose level was a significant negative prognostic factor. It is concluded that there is higher relapse rate, poor prognosis in adult ALL in comparison with children. In order to decrease the relapse rate and prolong the EFS, individual therapeutical regimens and prophylaxis of complicating diseases should be applied to ALL patients.

[A study on the cell differentiation induced by tanshinone IIA and its molecular mechanism in retinoic acid: resistant acute promyelocytic leukemia].

OBJECTIVE: To investigate retinoic acid-resistant acute promyelocytic leukemia (APL) cell differentiation induced by tanshinone IIA (Tan IIA) and its molecular mechanism. METHODS: NB4 cells treated with 0.5 mg/L Tan IIA was regarded as positive control. After in vitro incubation of MR-2 cells with Tan IIA at the concentration of 1.0 mg/L for 4 days, the cell differentiation was observed by growth status, cytomorphology, and nitroblue tetrazolium test. Cell cycle, membrane cluster differentiation (CD) antigens (CD(33), CD(11b)) and expression of some oncogene (c-myc, c-fos, p53 and bcl-2) were analysed by flow cytometry. RESULTS: The growth of MR-2 and NB4 cells was inhibited after Tan IIA treatment, the inhibition rate were 73.5% and 67.7% respectively (P < 0.01, P < 0.01) without significant difference. After Tan IIA treatment, MR-2 and NB4 cells were induced to undergo morphological differentiation, which exhibited small cell bulk decreased nucleus/cytoplasm proportion, rough chromatin, disappearance of nucleolus and formation of azurophil granules and anomalous nucleus. MR-2 cells could be induced to metamyelocyte while NB4 could be induced to band form. NBT reduction of MR-2 and NB4 cells treated with Tan IIA showed that positive cells accounted for (95.30 +/- 0.76)% and (93.20 +/- 1.04)% respectively; but the positive rate of either group of the treated positive cells was significantly higher than that of untreated, being (3.50 +/- 1.32)% and (2.80 +/- 0.29)% respectively (P < 0.01). Flow cytometry showed that the expression of CD(33) was reduced, while that of CD(11b) was increased. The quantity of treated cells in G(0)/G(1) phase increased but that in S phase decreased. The proliferous index was also decreased. After treated with Tan IIA, the expressions of anti-oncogene p53 and c-fos were up-regulated while those of oncogene bcl-2 and c-myc were down-regulated (P < 0.01). CONCLUSIONS: 1.0 mg/L Tan IIA could inhibit proliferation of MR-2 cells and induce differentiation of MR-2 cells into mature granulocyte, the effectivity was the same as 0.5 mg/L Tan IIA treated NB4 cells. Its possible molecular mechanism might be related to modulation of oncogene expressions associated proliferation and differentiation as well as inhibition of DNA synthesis. Tan IIA probably can be applied to treat the patients with APL, particularly to the relapsed and drug resistant patients with broad prospect.