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To determine the effect of pachymic acid (PA) on proliferation, cell cycle, and invasion in human ovarian carcinoma cell lines HO-8910 and explore some possible mechanisms, HO-8910 cells was treated with different concentrations of PA (0.5, 1, 2 μM). CCK-8 assay, propidium iodide staining, was applied to measuring the growth inhibiting rates of HO-8910 cells. Cell cycle was measured by flow cytometry. In addition, the activity of PA against HO-8910 cells invasion was evaluated in transwell assay. Western blot detected the proteins expression of E-cadherin, β-catenin and COX-2 of different groups treated with PA in different concentrations (0.5, 1, 2 μM) for 48 h. Our results showed that PA could effectively inhibit the in vitro growth of HO-8910 cells in dose-dependent manners in 72 h, suppressed migration and invasion of HO-8910 cells in concentration-dependent manners at 24 h, caused the increased accumulation of G1 phase cells, and caused down-regulation of β-catenin and COX-2 and up-regulation of E-cadherin expression level. Taken together, it could conclude that PA might inhibit proliferation and invasion of ovarian carcinoma cell through decreasing β-catenin and COX-2 expression and increasing E-cadherin expression.

This study was aimed to explore the expression level and correlation of MCL-1 and miR-29a in extranodal NK/T-cell lymphoma (ENKTCL) tissue. Maxvision immunohistochemistry technique and real time fluorescent quantitative PCR were used to detect the expression level of MCL-1 and miR-29a in tissue of 20 patients with ENKTCL and 10 patients with proliferative lymphadenitis, respectively. The results showed that the expression of MCL-1 protein were higher in patients with ENKTCL than that in patients with proliferative lymphadenitis, but there were no significant correlation between MCL-1 overexpression and age, sex, Ann Arbor stage and International Prognostic Index (IPI), respectively. Correlation analysis indicated that there was significant negative correlation between miR-29a expression and MCL-1 expression (r = -0.59, P = 0.016). It is concluded that miR-29a may target MCL-1 gene, regulate its expression, then participate in tumorigenesis and development of ENKTCL.

This study was aimed to construct miRNA sponge targeting miR-20a and to establish a stable cell line Jurkat-S, paving the way for further research on function of miR-20a and application of RNAi in gene therapy. One pair of two-repeated oligonucleotide sequences containing bulged sites that are mispaired opposite miR-20a positions 9-12 was designed and synthesized with enzyme cutting sites. The annealed oligonucleotide fragments were subcloned into pCDNA3.0-L expressing vector. After double-enzyme cutting, the vector was ligated to the annealed oligonucleotide fragments again. Enzyme cutting and luciferase activity assay were performed for identification after four repeats. Then the ligated fragment was subcloned to lentivirus expressing vector. Virus particles were collected after the control or sponge vectors were co-transfected with the psPAX2 packaging plasmid and the envelope plasmid pMD2.G into HEK-293T cells using Lipofectamine 2000. The Jurkat cells were transfused with recombinant lentivirus-transfusing units plus 6 µg/ml of Polybrene. Real-time PCR and Western blot were used to detect the mRNA and protein expression of P21 and E2F1 after lentivirus transfusion respectively. As a result, luciferase activity assay demonstrated that the sponge targeting miR-20a was constructed successfully and the virus was packaged in 293T. The titer of virus was 5×10(7) TU/ml. Stable transfected Jurkat-S cell line was established. As was expected, the mRNA and protein level of P21 and E2F1 was upregulated significantly in Jurkat-S cells. It is concluded that the miR-20a sponge is constructed successfully, and Jurkat-S stable cell line is established, in which the expression of miR-20a is inhibited stably.

This study was aimed to construct lentivirus-mediated shRNA expression vector targeting Bmi-1 and establish a stable cell line U266-li, so as to pave the way for further research on function of Bmi-1 and application of shRNA to gene therapy. One pair of oligonucleotide sequences targeted at human Bmi-1 mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pLVTHM vector. Virus particles were collected after the control or shRNA vectors were co-transfected with the psPAX2 packaging plasmid and the plasmid pMD2.G was enveloped into HEK-293T cells by using Lipofectamine2000. The U266 cells were transduced with 5 × 10(6) recombinant lentivirus-transducing units plus 6 µg/ml of polybrene. Real-time PCR and Western blot were used respectively to detect the expression of Bmi-1 and P14 after lentivirus transduction. DNA sequencing demonstrated that the lentivirus RNAi vector of Bmi-1 was constructed successfully and the virus was packaged in 293T cells. The titer of virus was 5 × 10(7) TU/ml. Stable transfected U266 cell line was established. As was expected, the mRNA and protein levels of Bmi-1 was reduced significantly in U266 cells after lentivirus transduction, whereas the mRNA and protein levels of P14 was upregulated. It is concluded that the lentiviral RNAi vector of Bmi-1 is constructed, and U266 stable cell line is established.

This study was aimed to explore the synergistic effect of 2-methoxyestradiol (2-ME2) and bortezomib (Bor) on the proliferative inhibition and apoptosis of U266 cell line and its possible mechanism. The cells were treated with 2-ME2, Bor alone and 2-ME2 combined with Bor, respectively. The cell viability and proliferative curve were detected by CCK8, the cell apoptosis was detected by caspase 3/7 activity test, cell cycle status was analyzed by flow cytometry, and real-time PCR was used to detect the mRNA expression of P21, BAX and BCL-2. The results showed that compared with cells treated with 2-ME2 or Bor alone, the proliferative potential of cells in combination group was significantly inhibited (p0.05), and apoptosis rate markedly increased (p0.05), cell cycle was arrested at G(1)-S phase, the mRNA expressive level of P21 and BAX increased, while the expression of BCL-2 decreased. It is concluded that 2-ME2 combined with Bor synergistically inhibits cell proliferation and induces apoptosis in U266 cell line. The possible mechanism may be associated with its effect of up-regulating P21 and BAX expressions.

Primary human endometrial cells were exposed to hypoxia preconditioning (HPC), HPC-hypoxia, and hypoxia conditions, and then endometrial tissue treated with ischemia preconditioning (IPC) was transplanted onto the chick embryo chorioallantoic membrane to investigate the role of slight ischemia of endometrium in the pathologic process of endometriosis. IPC up-regulated vascular endothelial growth factor expression and decreased apoptosis of endometrial cells, thus facilitating the endometrial fragments' ectopic implantation.

To investigate the inhibitory effects of Pomegranate peel extracts (PPE) on experimental prostatitis rats.The Prostatitis model was established by injection of carrageenan and Staphylococcus aureus in rat prostate. After 9 days' treatments of PPE, the rats were sacrificed for measurement of the number of white blood cell (WBC), the density of lecithin, the levels of Zn and malondialdehyde (MDA) of prostate. The antioxidant capacity was measured by oxygen radical absorbance capacity (ORAC) assay. The levels of NO and the activity of acid phosphatase (ACP) of plasma were also measured.PPE increased the levels of ORAC in plasma and the density of lecithin and the levels of Zn, decreased the levels of MDA of prostate and the activity of ACP and the number of WBC and adjusted the levels of NO in plasma compared with the prostatitis model group (P0.05 or P0.01).PPE can markedly improve the protective function of oxidation resistance, it may involve its regulatory action on anti-oxidation.

A study was carried out taking low temperature domestic wastewater as target in laboratory, the composition of bacteria in activated sludge was analyzed and the degradation kinetics of organics was investigated. Six preponderant psychrotrophs were selected out, belonging to Zoogloea, Aeromonas, Flavobacterium, Micrococcus, Bacillus and Pseudomonaus, respectively. Results show that TTC-dehydrogenase activity of the psychrotroph is 25.44 mg/(L x h) which is 20.5 times more than that of ordinary activated sludge at 4 degrees C. The efficiency of COD biodegradation by psychrotrophs at low and normal temperature is 80.9% and 73.4%, respectively. Mesophilic bacteria almost lost their activity at low temperature. Kinetic analysis shows that biodegradation of organics by psychrotrophs at low and mesothermal temperature as well as mesophilic bacteria at mesothermal temperature are in accordance with the model of first-order reaction. Psychrotrophs, which assure the removal efficiency of organic pollutants at low and normal temperature, could keep the uniform reaction velocity as the mesophilic bacteria and also adapt wide ecological amplitude of temperature. Efficient psychrotrophs were immobilized on soft polyurethane foams which acted as carriers in the experiment at 4 degrees C, as a result of that, the removal efficiency of COD was increased about 18% higher than that of mobilized ones at low temperature, the biodegradation kinetics of COD by immobilized psychrotrophs also followed the first-order reaction model. With glucose in water as the source of nutriment, the reaction velocity of immobilized bacteria is 2.35 times higher than that of the mobilized ones. By the immobilized psychrotrophs biodegradation of varied nutriment, the effluent could achieve the first-degree B of pollutants discharge standard for municipal wastewater treatment plant.

To establish an HPLC method for the content determination of epigoitrin in Radix Isatidis and its preparation, and to provide valuable data for quality control of Radix Isatidis and its preparation.The samples were separated on a ZORBAX SB-C18 (4. 6 mm x 150 mm, 5 microm) column with the mobile phase of acetonitrile-water-phosphoric acid-triethylamine (8.50 : 90.72 : 0.73 : 0.05) in the flow rate of 0.7 mL x min(-1). The detection wavelength was set at 245 nm. Column temperature was 30 degrees C.The linear range of epigoitrin was 0.0204-0.3060 microg (r = 0.9998), and the average recovery was 98.99% with the RSD was 1.31% (n = 9).The method for quantitation of epigoitrin in Radix Isatidis and its preparation was accurate and reliable, which can be used to evaluate the quality of Radix Isatidis and its preparation.

To observe the protective effects of pomegranate peel extracts on oxidative stress in restrained mice.The liver injury model was established under 18 h restraint stress in mice. The protective effects were evaluated by assessing alanine aminotransferase (ALT) GSH and Malondialdehyde (MDA) levels in plasma and antioxidant capacities of liver were measured by using oxygen radical absorbance capacity (ORAC) assay.Activities of ALT and levels of MDA were observed to decrease on pomegranate peel extracts treated mice as compared with restraint stress control group and antioxidant capacity in liver homogenization and the levels of GSH in plasma significantly increased as compared with restraint stress.The results indicate that pomegranate peel extracts exert protective effects on oxidative stress in mice loaded with restraint stress, which may be related to its free radical scavenging activity and lipid peroxidation inhibitory effect.

Zinc hydroxide prepared in laboratory was used as catalyst in catalytic ozonation of trace p-chloronitrobenzene (pCNB) in water. The catalytic mechanism was deduced through the effect of radical inhibitor t-BuOH on the reaction. Furthermore, affecting factors of catalytic activity were investigated. Results show that zinc hydroxide has excellent property of catalytic ozonation. Compared to ozonation alone, the catalytic removal rate of pCNB dissolved in distilled water increases by 51.3 percents at reacting time of 20 min. The catalysis, which is influenced by density of hydroxyl groups combined on the surface of hydroxide, follows a hydroxyl radical reaction pathway. The removal gets better with increasing dosage of catalyst. Zinc hydroxide was used for 3 times without obvious changes of pCNB removal rate. Bicarbonate in water and phosphate in buffer solution could greatly depress its catalytic activity. The optimal catalysis is achieved at solution pH = 7.