Objective To analyze the regulation of NEF3 expression in SH - SY5Y after RA induction. Methods The expression of NEF3 and Brg1 genes was individually detected by realtime RT - PCR and Western blot analyses. Then, the CAT reporter driven by 5' flanking region ofNEF3 gene (pBLCAT3 -2. 8k - NEF3) was co - transfccted with eukaryotic expression plasmids of pBJ5 - Brgl or pBJ5 - Brg1 - NTP into SH - SY5Y respectively. Promoter activity of the NEF3 gene was detected by competitive RT - PCR assay. Results Similar changes in the mRNA expression of NEF3 and Brg1 and that of the protein expression of Brgl were detected in Sh - SY5Y cells af-ter RA treatment for 12 -24h. Wild type Brgl can promote the expression of pBLCAT3 -2.8k -NEF3 promoter after RA induction, but not mutant Brg1. Conclusion Brgl helpes regulating the expression of NEF3 gene possibly via changes in chromatin conformation in the SH -SY5Y cells after RA induction.

AIM:Xilei Powder,a traditional Chinese prescription,has been used to treat wounds for hundreds of years,but the mechanism has not been fully understood.METHODS:The effects of Xilei Powder on fibroblast proliferation,collagen accumulation,matrix metalloproteinases-2,9 ( MMP-2,9 ) activities and tissue inhibitor of metalloproteinase 1 (TIMP-1) production were investigated by MTT,chloramine T method,gelatin zymography and enzyme-linked immunosorbent assays ( ELISA),respectively.RESULTS: The aqueous extract of Xilei Powder significantly promoted fibroblasts proliferation in a time and concentration manner,the population doubling time (125 μg/mL) was 33.8 h,it also significantly (P ＜0.05 ) promoted collagen production.Both of the aqueous and alcoholic extracts could significantly ( P ＜ 0.05 ) increase MMPo2 activity,and also very significantly ( P ＜ 0.01 )promote TIMP-1 production.CONCLUSION: Xilei Powder could promote fibroblasts proliferation,collagen and TIMP-1 production,this might be parts of mechanism to promote wound healing.

AIM:Xilei Powder,a traditional Chinese prescription,has been used to treat wounds for hundreds of years,but the mechanism has not been fully understood.METHODS:The effects of Xilei Powder on fibroblast proliferation,collagen accumulation,matrix metalloproteinases-2,9(MMP-2,9)activities and tissue inhibitor of metalloproteinase 1(TIMP-1)production were investigated by MTT,chloramine T method,gelatin zymography and enzyme-linked immunosorbent assays(ELISA),respectively.RESULTS:The aqueous extract of Xilei Powder significantly promoted fibroblasts proliferation in a time and concentration manner,the population doubling time(125 ?g/mL)was 33.8 h,it also significantly(P

Bone Neoplasms ; Humans ; Ilium ; Paraganglioma ; Sacrum

Objective To clarify the possible gene mutations in luteinizing hormone(LH) receptor gene in a boy with LH independent precocious puberty and probe the mechanism the of diseases caused by LH receptor activating mutations. Methods ( 1 ) Describe the clinical manifestations and laboratory data in a 5-year-old boy with LH independent precocious puberty. (2) Peripheral leukocytes were collected from the proband, his parents and other 20 normal puberty developed males. PCR and direct DNA sequence of 11 exons in LH receptors gene were conducted. Results (1) The proband was diagnosed to have LH independent precocious puberty according to the clinical symptoms and the laboratory tests. (2) A germ-line heterozygous point mutation in the 11 exon of LH receptor gene was found in the proband and his mother:c1193 T→C leading to amino acid change with M398T, which causes consecutively an activation of the LH receptor. (3) Other nucleotide changes in the proband and other normal males include c935 A→ G (N312S) and c1065 T→C(same sense mutation). Conclusions (1) A germ-line heterozygous point mutation in the LH receptor gene with M398T leads to consecutively activation of the LH receptor and LH independent precocious puberty. (2) The same point mutation does not have any influence on the puberty development, menstruation and productive functions of the proband's mother. (3) The LH receptor gene has possible polymorphism in the Han ethnic population.

OBJECTIVE To establish an in vitro test method and to evaluate the genotoxicity of chemicals using primary cultured mouse hepatocytes and the changes in phosphorylated histone H2AX(γH2AX)expression levels to provide a more reliable marker of the identification of genotoxicity. METHODS Hepatocytes were isolated from BALB/c mice by an improved two-step collagenase diges?tion method and then cultured in sandwich configuration. The primary cultured hepatocytes were treat?ed with various concentrations of four known genotoxic agents bleomycin(BLM),benzo(a)pyrene〔B (a)p〕,styrene and styrene-7,8-oxide(SO)within the range of 40 μmol · L-1 and two non-genotoxic agents azathioprine(Aza)and ciclosporin A(CsA)at different time points within 24 h. The cytotoxicity induced by these toxicants was assessed by CCK-8 assay. Then,the changes in γH2AX expression levels in treated cells were determined by flow cytometry. RESULTS The four genotoxic agents could be detected and two non-genotoxic agents could not be detected by this method. The γH2AX expression level was the highest when hepatocytes were exposed to BLM and SO for 3 h,or B(a)p and styrene for 6 h(P<0.01). The production of γH2AX was 25.67,18.36,12.43 and 14.25 for the four types of genotoxic agents,respectively,and was approximately 19,13,9 and 11 times that of the vehicle control group(P<0.01)at the optimum time point and concentration. There was a significant positive corre?lation between the indicated concentrations of genotoxic chemicals and γH2AX expression levels(P<0.01). In addition,the production ofγH2AX indicated no marked increase in two non-genotoxic agents such as Aza and CsA in comparison with the control group. CONCLUSION This test method can effec?tively distinguish genotoxic agents from non-genotoxic agents,and direct genotoxic agents from indirect genotoxic agents in the absence of S9. γH2AX might be a reliable marker for the identification of the potential genotoxicity of chemicals.

OBJECTIVE To find the infla mmation bio markers induced by coke oven e missions (COE),we investigated the changes of T helper 17 (Th17 )cytokines in hu man bronchial epithelial (16HBE)cells.METHODS 16HBE cells were exposed to organic extracts of COE collected fro m co-king plant at the concentrations of 5,10 and 20 mg·L -1 for 24 h or 5 d to establish short-term and long-term cell models,respectively.Cell viability was measured by MTT assay and infla mmatory da mage was assessed by lactate dehydrogenase assay (LDH).The cytokines in culture supernatant sa mples was detected by co mmercial hu man Th17 cytokine panel kit.RESULTS COE Can induce infla mmation in COE 20 mg·L -1 group and no expression on IL-17 F and IL-1 β.The concentration of IL-10 was 1 .25 ± 0.54,1 .39 ±0.13 and (1 .90 ±0.73)pg·mL -1 in COE 5,10 and 20 mg·L -1 group showing good con-centration-effect relationship (r=0.98,P <0.05 ).IL-23 expression was found only higher at 10 and 20 mg·L -1 and the concentrations were 3.38 ±3.90 and (1 .74 ±2.00 )pg·mL -1 ,respectively.In 16HBE cells treated by COE for 5 d,elevated expression of IL-17A was found in COE 5 and 10 mg·L -1 group,and there was statistically sigificant difference between COE 10 mg·L -1 and DMSO group (P<0.05).Elevated concentration of IL-17F of 10.2 ±1 1 .78 and (6.79 ±7.84)pg·mL -1 was found in COE 5 and 10 mg·L -1 group.The concentration of IL-10 was 1 .71 ±0.02,1 .49 ±0.25 and (2.82 ± 0.33)pg·mL -1 in COE 5,10 and 20 mg·L -1 group,respectively.We found increased IL-1 βexpression with concentration of 2.72 ±0.62,2.25 ±0.33 and (0.93 ±0.21 )pg·mL -1 in COE 5,10 and 20 mg·L -1 group with negative dose-response relationship.We also found more elevated TNF-αlevels in the 5 d than in the 24 h model with no COE specific relationship.CONCLUSION COE induces expression changes of Th17 cytokines profile in 16HBE cells,including IL-23 and IL-1 βfor early and long-term infla mmation,respectively.IL-10 may be a candidate marker for population study on COE induced infla mmatory injury.

To explore the influence of calculus bovis on the function of primary cultured mice oral fibroblasts, we determined the effects of calculus bovis on the fibroblast proliferation, collagen production, matrix metalloproteinases-2, -9 activities and tissue inhibitor of metalloproteinase-1 production by MTT assay, chloramine T method, gelatin zymography and enzyme-linked immunosorbent assays respectively. The results showed that calculus bovis could significantly inhibit the proliferation of fibroblasts and collagen synthesis in a concentration dependent manner, could significantly (P<0.05) suppress matrix metalloproteinases-2 activity and very significantly (P<0.01) inhibit the production of tissue inhibitor of metalloproteinase-1. In conclusion, the major function of calculus bovis in the process of ulcer healing is not to promote tissue regeneration, the mechanism that calculus bovis inhibits collagen synthesis may be partly due to its ability to very significantly (P<0.01) suppress the production of tissue inhibitor of metalloproteinase-1.
Animals ; Cattle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cholelithiasis ; chemistry ; veterinary ; Collagen ; drug effects ; metabolism ; Fibroblasts ; cytology ; physiology ; Materia Medica ; pharmacology ; Mice ; Mice, Inbred BALB C ; Mouth Mucosa ; cytology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism

The tumor multidrug resistance reversal effect of NPB304, a novel taxane, was studied. MTT assay was used to determine the IC50 of chemotherapy drugs. Western blotting assay was applied to analyze the expression of P-glycoprotein (P-gp). The effect of compounds on the P-gp function and P-gp ATPase activity was determined by rhodamine 123 (Rh123) accumulation assay and analysis kit, respectively. Molecular docking was employed to predict the binding force between compounds and P-gp. Transmembrane transport of NPB304 was analyzed using MDCK II and MDR1-MDCK II cell model. NPB304 displayed multidrug resistance reversal effect on KBV cells and MCF-7/paclitaxel cells, NPB304 collaborative with P-glycoprotein (P-gp) inhibitors verapamil enhanced the reversal activity, specifically, 10 μmol x L(-1) verapamil in combination with paclitaxel reversed resistance by 56.5-fold, while combined with NPB304 increased the reversal fold; NPB304 synergistically increased Rh123 accumulation in the resistant cells when combined with verapamil, and NPB304 at 0-1 μmol x L(-1) enhanced the ATPase activity activated by verapamil was observed. NPB304 existed the hydrophobic interactions with the TM regions of P-gp, and the binding force between NPB304 and the A chain of the TM region was stronger. P-gp ATPase activity assay demonstrated NPB304 at lower concentrations (0-1.5 μmol x L(-1)) could activate the P-gp ATPase, playing a role on inhibition of P-gp function. However, NPB304 did not have an obvious feature of P-gp substrate. NPB304 exerted itself and synergy with verapamil activity on reversing tumor resistance via inhibiting the P-gp function.

Objective To compare selenium content in hair samples of people in Kaschin-Beck disease (KBD) areas and control areas in Shaanxi Province two years after stopping the selenium salt prevention,and to provide a scientific basis for development of targeted prevention measures and for decision-making.Methods In July 2014,the four KBD counties of Yongshou,Yuyang,Linyou and Nanzheng were selected as survey counties,meanwhile,four non KBD counties of Wugong,Mizhi,Qishan and Chenggu were selected as rural control groups and Lianhu District in Xi'an City was selected as a urban settlement control county.Four villages (communities) were selected as monitoring sites according to four directions as east,west,south and north in each county.In each monitoring site,hair samples of 8 children aged 7-12 years old (gender balanced) and 8 adults over the age of 16 (gender balanced) were selected to determine hair selenium.Samples were prepared by wet digestion method,the content of selenium was determined by 2,3-diaminonaphthalene fluorescence method.Results A total of 576 hair samples were collected.The average hair selenium in each monitoring site was more than 0.20 mg/kg.Hair selenium was compared in KBD areas,rural non KBD areas,and urban non KBD areas,the differences were not statistically significant [(0.35 ± 0.18),(0.41 ± 0.28),(0.46 ± 0.19) mg/kg,F =1.544,P ＞ 0.05].In KBD areas and non KBD areas,there were 45 and 45 people with selenium content ＜ 0.20 mg/kg,accounting for 17.58％ and 14.06％;47 and 25 people with selenium content 0.20-＜ 0.25 mg/kg,accounting for 18.36％ and 7.81％;113 and 159 people with 0.25-＜ 0.50 mg/kg,accounting for 44.14％ and 49.69％;51 and 91 people with ≥0.50 mg/kg,accounting for 19.92％ and 28.44％.Hair selenium content of children aged 7-12 and adults was compared in KBD areas,rural non KBD areas,and urban non KBD areas [children:(0.43 ± 0.35),(0.38 ± 0.19),(0.50 ± 0.16) mg/kg;adults:(0.32 ± 0.17),(0.38 ± 0.19),(0.42 ± 0.21) mg/kg],the differences were not statistically significant (F =2.131,1.789,P ＞ 0.05).Hair selenium content was compared in different gender in KBD areas,rural non KBD areas,and urban non KBD areas [male:(0.35 ± 0.18),(0.44 ± 0.33),(0.52 ± 0.15) mg/kg;female:(0.35 ± 0.19),(0.38 ± 0.22),(0.41 ± 0.21) mg/kg],the differences were not statistically significant (F=1.598,1.790,P ＞ 0.05).Conclusion Two years after stopping the selenium salt prevention in Shaanxi Province,the selenium of population in Kashin-Beck disease areas in Shaanxi Province exceeds the human health threshold (0.20 mg/kg),and most people are in the medium or high levels of selenium,and are close to the levels of selenium in non-endemic areas.