Batch and continuous fermentation were adopted to investigate the effect of specific growth rate and amino acid components on RNA accumulation in Candida tropicalis ATCC 20408 in fermentation medium ( FM), yeast peptone dextrose medium (YPD), molasses fermentation medium ( MFM) and FM without corn steep liquor. The data showed that obvious differences in intracellular RNA accumulation were observed at different cell growth phases in bath fermentation prosess, and RNA level reached 11. 8% (g-RNA /g-DCW) during exponential phase, and only 6.9% during stationary phases. It was also found that intracellular RNA accumulation increased with the increase of specific growth rate in continue fermentation prosess, and the highest RNA level reached 15. 6% with the glucose conversion rate of 42. 8% at the dilution rate of 0. 5 h-1. Furthermore, the data showed that RNA lever was notably increased in batch fermentation process when amino acids or peptone was added into the fermentation medium containing no corn steep liquor. Taken together, it was reported for the first time that specific growth rate and amino acid components plays a leading role on the intracellular RNA accumulation in C. tropica lis, and specific growth rate is more important.

Objective To explore the cytotoxic effects and mechanisms of Aflatoxins B1(AFB1) on the African green monkey kidney cells (Vero E6).Methods After incubation of Vero E6 cells with AFB1,cellular toxicity was assessed by CCK-8 assay and LDH release rate.Annexin V/PI double staining and DAPI staining was used to detected cell apoptosis.Results AFB1 has significant toxic effects on Vero E6 cells.Cell viability was less than 10% when treated by 200 μmol/L AFB1 for 48 h.LDH release rate was increased significantly with the increase of AFB1 concentration.Externalization of cell membrane phosphatidylserine was observed by Annexin V/PI double staining.Nucleus fragmentation was detected by DAPI staining.Conclusion AFB1 exhibits significantly cytotoxic effects on Vero E6 cells by inducing cell apoptosis.

非编码RNA（ncRNA）是一类不具有编码蛋白功能的RNA转录本，其异常表达参与了肿瘤的发生与发展。ncRNA的 组织特异性表达使其具有鉴别肿瘤，甚至具有对肿瘤分型、分期的应用潜能。雌激素受体阴性（ER-）乳腺癌是指不表达ER的乳 腺癌，其治疗困难、预后极差、病死率居乳腺癌之首。ncRNA在ER-乳腺癌中存在差异表达，并可通过参与非雌激素激活通路在乳 腺癌的发生发展及预后过程中起重要调控作用。本文拟针对ncRNA中的微小RNA（miRNA）、长链非编码RNA（lncRNA）、环状 RNA（circRNA）在ER-乳腺癌中的作用展开综述，分析它们在临床中的应用价值， 为ER-乳腺癌的早期诊断及预后判断提供新的 分子标志物和监测手段。

OBJECTIVETo investigate the suppressive effect of resveratrol on growth of U251 human glioma cells and its correlated mechanism.

METHODU251 human glioma cells were treated with resveratrol at various concentrations, MTT assay was used to determine the inhibitory rate of cell proliferation, FCM to detect the cell apoptosis, the expressions of Bcl-2, Bcl-XL, STAT3 and CyclinD1 were analysed by immunohistochemistry and Western blot to examine the expression of Bcl-2, Bcl-XL, STAT3, CyclinD1, Caspase-3 and Bax.

RESULTAfter treatment with resveratrol, MTT assay showed the growth of U251 cells was inhibited in dose-dependent and time-dependent manners, apoptosis of cells advanced stage was built up, immunohistochemical staining displayed decreased the expression of Bcl-2, Bcl-XL, STAT3 and CyclinD1 and Western blot showed that resveratrol decreased the expression of Bcl-2, Bcl-XL, STAT3 and CyclinD1, and built up Bax and Caspase-3.

CONCLUSIONIt is possible that downregulated the expression of Bcl-2, Bcl-XL, but upregulated Bax and Caspase-3, and the indication was obviously in dose-dependent and time-dependent manners.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Glioma ; drug therapy ; metabolism ; Humans ; Immunohistochemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Stilbenes ; therapeutic use ; bcl-2-Associated X Protein ; metabolism ; bcl-X Protein ; metabolism

OBJECTIVETo investigate the intestinal absorption characteristics of gastrodigenin.

METHODIn vitro everted gut sac model and in situ rat single-pass intestinal perfusion model were used to evaluate the absorption characteristics of gastrodigenin in the different intestinal segments. The concentrations of gastrodigenin in the samples were determined by Ultra Performance Liquid Chromatography (UPLC) method, and the relevant absorption parameters were calculated.

RESULTIn the everted gut sac tests, no significant difference of absorption among the four segments was observed. A positive correlation was found between drug concentration and the accumulated absorption amount (Q). At the concentration of 400 mg x L(-1), the Q of gastrodigenin in the duodenum, jejunum, ileum and colon were 224.33, 225.81, 233.18 and 189.25 microg, respectively. The in situ rat single-pass intestinal perfusion tests showed that there was also no significant difference of absorption among the four segments. The absorption rates (A) of gastrodigenin in the duodenum, jejunum, ileum and colon were 45.8%, 48.39%, 47.00%, 54.35%, respectively.

CONCLUSIONGastrodigenin can be well absorbed via passive diffusion in the intestine. The absorption rates of gastrodigenin in the different intestinal segments show no regioselectivity.

Animals ; Benzyl Alcohols ; pharmacokinetics ; Chromatography, Liquid ; methods ; Intestinal Absorption ; Intestines ; metabolism ; Male ; Medicine, Chinese Traditional ; Models, Biological ; Perfusion ; methods ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effects of histone deacetylase inhibitors trichostatin A (TSA) on acute lung injury in septic mice.Methods Septic mice model was induced by cecal ligation and puncture(CLP).Ninty male BALB/c mice of clean grade were randomly(random number) divided into six groups(n=15),namely sham operation group,CLP group,CLP+DMSO group,CLP+TSA 1 mg group,CLP+TSA 5 mg group,and CLP+TSA 10 mg group.TSA(1 mg/kg,5 mg/kg,10 mg/kg) was administrated 12 hours before operation by intraperitoneal injection.And mice in sham group were only treated with laparotomy without CLP,and 24 h later,all survived mice were sacrificed to obtain specimens.ELISA method was employed to detect the concentrations of TNF-α and IL-1β in BALF.The lung wet/dry ratio was calculated.Histopathology changes of lung tissues were observed under light microscope.Lung tissue cell apoptosis was detected by TUNEL method.Caspase-3,Caspase-9 and CytC were assayed by Western blotting.The survival rate of mice in each group was calculated by additional 120 mice.Data were analyzed by SPSS 23.0.Statistical analyses were performed using independent sample t-test to compare between two groups or one-way analysis of variance test to compare among muhiple groups.The survival rate of mice was analyzed by univariate analysis using log-rank test.Results The lung W/D(P=0.021),the concentrations of TNF-α(P=0.000 1)and IL-1β(P=0.000 6)in BALF,puhnonary pathological change(P=0.001 6),lung tissue cell apoptotic index(P=0.000 9),the levels of apoptosis proteins (P＜0.05) in CLP group were higher than those in sham group,while survival rate (P=0.000 1) in CLP groups was lower than that in sham group.Compared with DMSO,the TSA significantly reduced the lung W/D,the levels of TNF-α.IL-1β in BALF,pathologic changes of lung tissue,lung tissue cell apoptotic index and the levels of apoptosis proteins in septic mice(P＜0.05).The increase in survival rate (P=0.007 2) associated with TSA(10 mg/kg)administration.Conclusion TSA exerts protective effects through attenuating pro-inflammatory cytokines and lung tissue cell apoptosis in sepsis induced acute lung injury in mice.

Objective To explore the antioxidant mechanism ofhistone deacetylase 2 (HDAC2) regulating Nrf 2 acetylation in lipopolysaccharide (LPS)-induced type Ⅱ alveolar epithelial cell injury.Methods The experiment was divided into two parts.The first part was the routine culture of type Ⅱ alveolar epithelial cells of mice.The cells were stimulated with different concentrations of LPS (10 ng/ mL,100 ng/mL and 1 000 ng/mL).CCK-8 was used to detect the cell activity at 0 h,6 h,12 h,24 h and 48 h,respectively.The second part:Alveolar epithelial cells of type Ⅱ were cultured and divided into the normal control group (control group),LPS group,HDAC2 lentivirus interference group (siRNA-HDAC2 group) and HDAC2 lentivirus overexpression group (LV-HDAC2 group).The expression of HDAC2 and Nrf2 were detected by Western blot,the acetylation of Nrf2 was detected by immunoprecipitation,and the stability of nrf2 was detected after actinidone action.The activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by chemical colorimetry.SPSS 23.0 statistical software was used.LSD-t test was used for comparison between two groups,and one-way ANOVA test was used for comparison among multiple groups.Results Compared with the control group,the expression of HDAC2 protein in the LPS group increased (t=5.974,P=0.027),the acetylation level of Nrf2 decreased (t=7.223,P=0.002),the Nrf2 protein level increased (t=2.929,P=0.043),the protein stability of Nrf2 increased,the SOD activity decreased (t=121,P＜0.01),and the MDA content increased (t=10.45,P=0.000 5).Compared with the LPS group,Nrf2 acetylation level decreased in the LV-HDAC2 group (t=1 1.29,P=0.000 4),Nrf2 protein expression increased (t=3.194,P=0.033),Nrf2 protein stability increased,SOD activity increased (t=4.678,P=0.009),and MDA content decreased in the LV-HDAC2 group (t=5.417,P=0.005 6).While the opposite trend was observed in the siRNA-HDAC2 group.Conclusion After LPS stimulation,oxidative stress of type Ⅱ alveolar epithelial cells was aggravated.HDAC2 could decrease the level of Nrf2 acetylation,increase the expression of Nrf2 protein,and alleviate LPS-induced oxidative stress.