[Overexpression of miR-31 regulates TLR4/NF-κB signaling pathway and apoptotic protein in colitis model mice].

Objective: To investigate the effects of miR-31 on TLR4/NF-κB signaling pathway and apoptosis-related proteins in dextran sulfate sodium (DSS) induced mouse colon colitis. Methods: ① Mouse model of colon colitis: 1% DSS was used to induce mouse ulcerative colitis (UC). Fourteen FVB non-transgenic mice were randomly divided into control group (n= 6), DSS group (n= 8), and 16 FVB miR-31 transgenic mice were randomly divided into miR-31 overexpression group (n= 8), miR-31 overexpression +DSS group (n= 8). DSS was dissolved in water and administered to mice by drinking water. The DSS group and miR-31+DSS group drank 1% DSS water in the first week, normal sterilized water in the second week, and 1% DSS water in the third week, after 5 weeks, the modeling was completed, then the colon tissues of the mice were collected. Western blot and IHC were used to detect the expressions of NF-κB p65, TLR4, Bax and Bcl-2 proteins in mouse colon tissue, TUNEL was used to detect apoptosis of mouse colon tissues. ② Cell culture experiments: Transfection of miR-31mimic and inhibitor by lipofectamine resulted in overexpression or knockdown of miR-31 in human colon epithelial cell line HCT 116 cells, each group was repeated three times and cells were collected 48 h later, Western blot was used to detect the expressions of NF-κB p65 and TLR4 protein. Results: ① In animal experiments, compared with the control group, the expression levels of NF-κB p65, TLR4 protein and apoptotic cell index in the DSS group and miR-31 overexpression group in mouse colon tissue were significantly increased (P＜0.05 or P＜0.01), and the Bcl-2 / Bax ratio was significantly reduced (P＜0.05 or P＜0.01); and compared with the DSS group, the expression levels of NF-κB p65, TLR4 protein and apoptotic cell index in the miR-31+DSS group were significantly increased (P＜0.01), while the Bcl-2/Bax ratio was significantly decreased (P＜0.01). ② In cell experiments, compared with the control group, the expression levels of NF-κB p65 and TLR4 protein in the over-expressed miR-31 group of HCT 116 cells were significantly increased (P＜0.05 or P＜0.01), the expressions of NF-κB p65 and TLR4 protein in miR-31 knockdown group were decreased (P＜0.05). Conclusion: miR-31 promotes the development of colitis by promoting TLR4/NF-κB signaling pathway and mediating apoptosis of intestinal epithelial cells.

[Effects of resveratrol on ulcerative colitis in mice and its mechanism].

OBJECTIVE: To investigate the anti-ulcerative colitis mechanism of resveratrol through regulation of Wnt/ß-catenin signaling pathway. METHODS: ①The experiment of ulcerative colitis induced by dextran sulfate sodium salt (DSS): 28 C57BL/6 mice were randomly divided into four groups including control group(n=7), DSS group(n=7), DSS+Resveratrol (DSS+Res) group(n=7) and Res group(n=7). The experiment lasted for 3 weeks. Ulcerative colitis of mice was induced by drinking DSS water and treated with resveratrol by intragastric administration. The mice were weighed daily and their activities and state of feces were recorded. After that, the mice were euthanized, the spleens were weighed, and the colonic length was measured.Hematoxylin-eosin staining (HE) was used to observe the pathological changes of the colon, and the expression of miR-31 in colonic tissue was detected by quantitative real-time PCR (qPCR). The expressions of ß-catenin and Cyclin D1 were measured by Western blot. ②In vitro experiment: HCT 116 cells were treated with resveratrol at 10 mg/ml, the expressions of ß-catenin, LDL receptor related protein-6 (LRP-6), frizzled-3 (FZD3) and c-Myc were detected. The expression of ß-catenin was also detected in HCT 116 cells transfected with miRNA-31 mimic and miRNA-31 inhibitor. RESULTS: ① The body weight was decreased in DSS group, the activity was decreased and blood stool appeared. The colonic length of mice was shortened, the spleen was enlarged and the tissue was damaged seriously in DSS group. While the above symptoms were improved after resveratrol treatment. ② Resveratrol inhibited the expressions of miRNA-31, ß-catenin and CyclinD1 in ulcerative colitis mice, and also down-regulated the expressions of ß-catenin, LRP-6, FZD3 and c-Myc in HCT 116 cells. After transfection of miRNA-31 inhibitor, the expression of ß-catenin was decreased in HCT 116 cells. CONCLUSION: Resveratrol suppresses DSS induced colitis by down-regulation of Wnt signal pathway. The down-regulation of Wnt signal may be related to miRNA-31.

[Determination of mineral elements in soybean from different producing areas by ICP-AES].

Soybeans from Jiangxi, Heilongjiang, Jilin and Anhui were digested by nitric acid. The contents of mineral elements K, P, Mg, Ca, Fe, Al, Zn, Mn and Na in four samples were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). The differences between samples were determined by Duncan's multiple-range test, and evaluations were based on a significance level of P < 0.05. The results show that soybeans are rich in mineral elements that human body needs. The contents of K, P, Mg, Ca and Fe are higher than the others, and the order from high to low does not change with the producing area. The order of Mn, Zn, Al, Na varied with the producing area. Producting area does not affect the contents of elements K, P and Mg significantly (P > 0.05). It can be concluded that the demands for major mineral elements K, P and Mg are stable in the growth of soybeans with the same variety. However, producing area has a great influence on the contents of elements Al, Ca, Fe, Na, Mn and Zn in soybean (P < 0.05). The content of Fe in Anhui soybean (70.93 mg x kg(-1)) and the content of Mn in Jiangxi soybean (32.69 mg x kg(-1)) are significantly higher than the other three samples. The results suggested that the contents of Al, Ca,Fe, Na, Mn and Zn were easily affected by growth conditions, such as water, soil and so on.

Ethyl 3-(4-methyl-benzene-sulfonamido)thieno[2,3-b]pyridine-2-carboxyl-ate.

The thieno[2,3-b]pyridine ring system of the title compound, C(17)H(16)N(2)O(4)S(2), is essentially planar, the amino and carbonyl groups being nearly coplanar with the heterocyclic ring system. There are two N-H⋯O hydrogen-bonding inter-actions involving the same N-H donor set and two different acceptors, one in an intra-molecular bond helping to fix the mol-ecular geometry and the other defining a dimeric structure around the symmetry centre at (0, , ).

Ethyl 3-amino-4H-thieno[2,3-b]pyridine-2-carboxyl-ate.

The mol-ecule of the title compound, C(10)H(10)N(2)O(2)S, is essentially planar, except for the ethyl group, which is twisted away from the carboxyl plane by -90.5 (3)°. In the crystal structure, mol-ecules are linked into a zigzag sheet propagating along the b axis by inter-molecular N-H⋯O and N-H⋯N hydrogen bonds.

Bis[4-(2-hydroxy-ethyl-amino)phen-yl] sulfone.

The title compound, C(16)H(20)N(2)O(4)S, exhibits a V-shape structure with a dihedral angle of 77.5 (11)° formed by the two benzenel rings. The mol-ecular packing is stabilized by intra-molecular and inter-molecular hydrogen bonds as well as π-π [3.738 (3) Å] and C-H⋯π inter-actions.

1-(1-Amino-4-bromo-9,10-dioxo-9,10-dihydroanthracen-2-ylcarbonyl)-1,3-dicyclohexylurea.

In the title compound, C(28)H(30)BrN(3)O(4), the molecules are linked by C-H...Br and N-H...O hydrogen bonds into one-dimensional chains, which are arranged into a three-dimensional network through a combination of C-H...O hydrogen bonds and two kinds of pi-pi interactions between the benzene rings of the anthraquinone units.

A zwitterion of 1,5-bis(2-hydroxybenzamido)-3-azapentane: a two-dimensional hydrogen-bonding network involving dimers.

The title compound, 2-{N-[2-(2-hydroxybenzamido)ethylammonioethyl]aminocarbonyl}phenolate, C18H21N3O4, crystallizes in a zwitterionic form as a result of intermolecular proton transfer and possesses a negatively charged phenolate group and a protonated amino group. The 2-hydroxybenzamide and 2-(aminocarbonyl)phenolate moieties attached to the two ends of the C-C-N-C-C backbone adopt a cis conformation in relation to this backbone. All N- and O-bound H atoms are involved in hydrogen-bond formation; the zwitterions are first linked into head-to-tail dimers, which are further organized into a two-dimensional network parallel to the crystallographic bc plane.