Daidzein affects proliferation and apoptosis in non-small cell lung cancer cells:role of p53 signaling pathway / 中国药理学通报

Aim To investigate the effects of daidzeinDD on the proliferation and apoptosis of non-small cell lung cancer cells,with a focus on the possible role of the p53 signaling pathway in this regard. Methods CCK-8 method and flow cytometry were used to detect the effects of soy isoflavone crude extract and DD on the viability and apoptosis of HELF and H1299 cells. Gene microarray was used to detect the changes in gene expression after treatment of H1299 cells with DD. GSEA and differential analysis were used to screen the major pathways and key genes. RT-qPCR and Western blot were performed to verify the differences in mRNA and protein expression of key genesp53 and CASP9 in the major pathways. After p53 inhibitor Pifithrin-α inhibited the expression of p53,the effect of DD on p53 mRNA and protein expression levels was examined,and the proliferative effect on H1299 cells was observed. Results Soy isoflavone crude extract and DD promoted proliferation and inhibited apoptosis of normal lung cells and inhibited proliferation and promoted apoptosis of lung cancer cells. p53 signaling pathway was significantly enriched in the DD-treated groupNES=1.78,P=0.000,and the expressions of p53 and CASP9 genes were found to be significantly up-regulated in the treated group. Compared with the control group,mRNA expression of CASP9 and p53 significantly increased in both HELF and H1299 cells treated with DDP<0.05,and p53 protein expression also increased in HELF cellsP<0.05. After inhibition of p53 expression,DD significantly increased the mRNA expression of p53 in H1299 and HELF cellsP<0.05 and also markedly increased the expression of p53 protein in H1299 cellsP<0.05,and it was observed that DD inhibited the proliferation of lung cancer cells. Conclusions DD inhibits the proliferation and promotes the apoptosis of lung cancer H1299 cells,and the mechanism mainly involves the p53 signaling pathway.

CHEMICAL REACTION MONITORING BY AMBIENT MASS SPECTROMETRY.

Chemical reactions conducted in different media (liquid phase, gas phase, or surface) drive developments of versatile techniques for the detection of intermediates and prediction of reasonable reaction pathways. Without sample pretreatment, ambient mass spectrometry (AMS) has been applied to obtain structural information of reactive molecules that differ in polarity and molecular weight. Commercial ion sources (e.g., electrospray ionization, atmospheric pressure chemical ionization, and direct analysis in real-time) have been reported to monitor substrates and products by offline reaction examination. While the interception or characterization of reactive intermediates with short lifetime are still limited by the offline modes. Notably, online ionization technologies, with high tolerance to salt, buffer, and pH, can achieve direct sampling and ionization of on-going reactions conducted in different media (e.g., liquid phase, gas phase, or surface). Therefore, short-lived intermediates could be captured at unprecedented timescales, and the reaction dynamics could be studied for mechanism examinations without sample pretreatments. In this review, via various AMS methods, chemical reaction monitoring and mechanism elucidation for different classifications of reactions have been reviewed. The developments and advances of common ionization methods for offline reaction monitoring will also be highlighted.

A pilot study of the therapeutic efficacy and mechanism of artesunate in the MRL/lpr murine model of systemic lupus erythematosus.

Recent evidence indicates that artesunate has immunomodulatory properties that might be useful for treating autoimmune disease. In this study, we conducted a pilot study and explored the effect and mechanism of artesunate on the treatment of systemic lupus erythematosus using an MRL/lpr murine model. MRL/lpr mice were divided into control, cyclophosphamide (CTX) and artesunate treatment groups. Blood was collected to measure serum levels of creatinine, antinuclear antibody (ANA) and anti-double-stranded DNA (anti-dsDNA) antibody. Twenty-four-hour urine was collected to measure levels of proteinuria. The concentration of monocyte chemotactic protein-1 (MCP-1) in serum and urine was measured. The expression of MCP-1 in kidney was detected by Western blot and immunohistochemistry assay, respectively. The expression of B cell activating factor (BAFF) in spleen was determined by real time-PCR and immunoblotting. We found that artesunate significantly increased the survival rate, body weight and blood leukocyte counts, and reduced the serum levels of ANA and anti-dsDNA antibody titer, 24 h urinary protein, and serum creatinine. Our results indicated that artesunate could decrease MCP-1, major pro-inflammation cytokine, in serum, urine and kidney. We also found that the level of BAFF, the major B cell activation factor, was decreased in artesunate treated MRL/lpr mice. Its efficacy was comparable with that of CTX in this study. Taken together, we have demonstrated that artesunate can inhibit the progression of disease and reverse the pathologic lesion of lupus nephritis.

[The effect of umbilical cord mesenchymal stem cell transplantation on interstitial pneumonitis in MRL/lpr mice].

OBJECTIVE: To investigate the effects of umbilical cord mesenchymal stem cell (UC-MSCs) transplantation on interstitial pneumonitis in MRL/lpr mice. METHODS: Twenty four 18-week-old MRL/lpr female mice were randomly divided into 3 groups: the single dose group received a single dose of UC-MSCs (1 x 10(6)) transfusion intravenously, the 3 dose group received 3 injections of UC-MSCs (1 x 10(6)) weekly for 3 weeks, and the control mice were treated with saline. Both the control and the treated mice were sacrificed at 29 weeks. The histopathology of the lungs were assessed by HE staining. RESULTS: In comparison to the control mice, UC-MSCs transplantation significantly attenuated interstitial pneumonitis in the MRL/lpr mice. The peribronchiolar lesion indices of the single dose treatment group (1.40 +/- 0.24) and the 3 dose treatment group (1.02 +/- 0.29) were significantly decreased as compared to the control group (1.95 +/- 0.35), q = 0.551, 0.937, all P < 0.01. The perivascular lesion index of the single dose treatment group (1.20 +/- 0.18) and the 3 dose treatment group (1.08 +/- 0.16) were also significantly reduced as compared to the control group (1.56 +/- 0.32), q = 0.360, 0.479, P < 0.05, P < 0.01. The inflammatory cell infiltration index of the control group (1.72 +/- 0.34) was significantly increased compared to the single dose treatment group (1.30 +/- 0.21) and the 3 dose treatment group (1.05 +/- 0.15), q = 0.417, 0.673, P < 0.05, P < 0.01. CONCLUSIONS: These findings indicate that UC-MSCs have a pleiotropic therapeutic effect on pneumonitis in MRL/lpr mice.

Transplantation of human bone marrow mesenchymal stem cell ameliorates the autoimmune pathogenesis in MRL/lpr mice.

Recent evidence indicates that mesenchymal stem cells (MSC) possess immunosuppressive properties both in vitro and in vivo. We previously demonstrated the functional abnormality of bone marrow derived MSC in patients with systemic lupus erythematosus (SLE). In this study, we aimed to investigate whether transplantation of human bone marrow derived MSC affects the autoimmune pathogenesis in MRL/lpr mice. We found that human MSC from healthy donors reduced the proliferation of T lymphocytes from MRL/lpr mice in a dose-dependent fashion. Two weeks after in vivo transfer of MSC, we detected significantly reduced serum levels of anti ds-DNA antibodies and 24 hour proteinuria in MRL/lpr mice as compared with control groups without MSC transplantation. Moreover, flow cytometric analysis revealed markedly reduced number of CD4(+) T cells while increased Th1 subpopulation in MSC group and MSC + CTX group when compared with controls. Histopathological examination showed significantly reduced renal pathology in MSC-treated mice. Immunohistochemical studies further revealed reduced expression of TGF-beta, FN, VEGF and the deposition of complement C3 in renal tissue after MSC and MSC + CTX treatment. Taken together, we have demonstrated that transplantation of human MSC can significantly inhibit the autoimmune progression in MRL/lpr mice.

The relationship between the beta-2-adrenergic receptor gene +46 Arg 16/Gly variant and serum level of low density lipoprotein-cholesterol in Kazakans of Xinjiang / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the distribution of beta-2-AR +46 A-->G variant in Kazakans of Xinjiang and the relationship of the variant with low density lipoprotein-cholesterol (LDL-C) level in this population.</p><p><b>METHODS</b>The genotypes of beta-2-AR gene Arg16/Gly variant were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique in 506 Kazakans with age from 30 to 69, and its distribution and relationship to LDL-C level were investigated.</p><p><b>RESULTS</b>(1) The frequencies of genotypes AA, AG, GG and alleles A, G of beta-2-AR +46 variant in this population were 0.310, 0.455, 0.235 and 0.538, 0.462 respectively, which were accorded with Hardy-Weinberg equilibrium. (2) The Gly16/Gly genotype had highest LDL-C level in the three genotypes, and were significantly higher than Arg16/Gly genotype (P < 0.05). (3) Comparing the effect of beta-2-AR gene +46 variant on serum lipid in males with females, we found that females with Gly16/Gly genotype had the highest level of serum LDL-C.</p><p><b>CONCLUSIONS</b>These data show that Gly16/Gly genotype of beta-2-AR gene +46 A-->G variant is associated with higher level of serum LDL-C in this population, especially in female.</p>

The relationship between the variants in 5' upstream core promoter A(-6)G and A(-20)C of angiotensinogen gene and essential hypertension in Kazakans of Xinjiang / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate whether the variants A(-6)G and A(-20)C of angiotensinogen (AGT) gene are involved in the pathogenesis of essential hypertension in Kazakans.</p><p><b>METHODS</b>T his case control study recruited 125 subjects with hypertension and 74 normotensive subjects from Kazakans of Xinjiang. Genomic DNA from leukocytes was analyzed for genetic variants A(-6)G and A(-20)C in 5' upstream core promoter of AGT gene by polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), restriction fragment length polymorphism (RFLP) and automatic sequencing.</p><p><b>RESULTS</b>(1)There were only A(-6)G and A(-20)C variants in the -164 to +73 region of Kazakans' AGT gene. (2) The distributions of genotypes AA, AG, GG at locus -6 of AGT gene showed significant difference between the hypertensive group (0.39, 0.45, 0.16) and the normotensive group (0.49, 0.49, 0.02; Chi2=8.56, P=0.014). There were evident differences in the frequencies of the -6A and the -6G allele of the two groups (0.62, 0.38 and 0.73, 0.27; Chi2=5.35, P=0.021). (3) No significant difference was observed in the distribution of genotypes AA, AC, CC at locus -20 of AGT gene between the hypertensive group (0.69, 0.26, 0.05) and the normotensive group (0.65, 0.32, 0.03; Chi2=2.42, P=0.30). There was no distinct difference in the frequencies of the -20A allele and the -20C allele of the two groups (0.82, 0.18 and 0.82, 0.18; Chi2=0, P=0.99). (4) No significant difference was found at the levels of systolic and diastolic blood pressure between the groups corresponding to genotypes at the loci -6 and -20 of AGT gene.</p><p><b>CONCLUSION</b>The results suggest that the polymorphism of A(-6)G in 5' upstream core promoter of the AGT gene may be involved in the pathogenesis of essential hypertension in Kazakans, while the A(-20)C variant may not play an important role in the etiology of essential hypertension in Kazakans.</p>