Self-assembled aggregations in Coptidis Rhizoma decoction dynamically regulate intestinal tissue permeability through Peyer's patch-associated immunity / 中草药·英文版

Objective: To investigate the dynamic regulation of self-assembled aggregations (SAA) in Coptidis Rhizoma decoction on the permeability of intestinal tissue and the mechanism underlying. Methods: The effects of SAA on berberine (Ber) absorption were respectively analyzed in an in situ intestinal perfusion model and in an Ussing Chamber jejunum model with or without Peyer's patches (PPs). The expression levels of ZO-1, Occludin and Claudin-1 were detected by immunofluorescence to evaluate the tight junction (TJ) between intestinal epithelium cells. The expression levels of T-box-containing protein expressed in T cells, signal transducers and activators of tranion-6, retinoic acid receptor-related orphan receptor γt and forkhead box P3 in PPs were detected by the reverse transcription-polymerase chain reaction and the secretions of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-17 (IL-17) and transforming growth factor-β (TGF-β) in PPs were evaluated by immunohistochemistry, to reflect the differentiation of T lymphocyte in PPs to helper T (Th) cell 1, Th2, Th17 and regulatory T (Treg) cell. To confirm the correlation between SAA in Coptidis Rhizoma decoction, PPs-associated immunity and intestinal epithelium permeability, SAA were administrated on an Ussing Chamber jejunum model with immunosuppressed PPs and evaluated its influences on intestinal tissue permeability and TJ proteins expression. Results: SAA in Coptidis Rhizoma decoction could dose-dependently promote Ber absorption in jejunum segment, with the participation of PPs. The dose-dependent and dynamical regulations of SAA on permeability of intestinal tissue and TJ proteins expression level between intestinal epithelium cells occurred along with the dynamically changed T lymphocyte differentiation and immune effectors secretion in PPs. The administration of SAA on immunosuppressed PPs exhibited dose-dependent PPs activation, inducing dynamic promotion on intestinal tissue permeability and inhibition on TJ proteins expression. Conclusion: SAA can improve the Ber absorption in small intestine, through the PPs-associated immunity induced dynamic regulation on intestinal tissue permeability and TJ proteins expression. These findings might enlighten the research of traditional Chinese medicine decoction.

Effects of Ubiquitination on the Expression of BCL6 Protein，Cell Proliferation and Apoptosis in K562/G01 Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the the effects of ubiquitin-proteasome system (UPS) on BCL6 protein level，proliferation and apoptosis of cell imatinib（IM）-resistant K562/G01 cells.@*METHODS@#Western blot was used to detect the expression of BCL6 in K562/G01 cells before and after treatment with protease inhibitor MG-132.The RT-PCR and Western blot respectively were used to detect the mRNA and protein expression levels of BCL6 and USP2 in K562/G01 cells treated with or without ML364 (a ubiquitin-specific protease USP2 inhibitor). The effects of IM alone or in combination with ML364 on proliferation and apoptosis of K562/G01 were analysed by CCK-8 method and flow cytometry.@*RESULTS@#After treatment with protease inhibitor MG132, the BCL6 protein level of K562/G01 significantly increased (P<0.05). The mRNA and protein expression level of ubiquitin-specific protease USP2 in K562/G01 cell line was higher than that in K562 cell line (P<0.05). After treatment of K562/G01 with USP2 protease inhibitor ML364, the expression levels of USP2 and BCL6 proteins were down-regulated simultaneously (P<0.05) . After combination of ML364 and IM, both the proliferation inhibitory rate and the apoptosis rate of K562/G01 cells significantly increased(P<0.05).@*CONCLUSION@#ML364 decreases the BCL6 protein stability in K562/G01 by inhibiting the USP2-mediated deubiquitination, and down-regulate the BCL6 protein experssion, thereby increases the sensitivity of drug-resistant cells to IM.

Mechanism of mTOR Pathway in K562 cell Apoptosis Induced by Homoharringtonine / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of homoharringtonine (HHT) on proliferation and apoptosis of CML cell line K562 cells and to explore its possible mechanism through mTOR pathway.</p><p><b>METHODS</b>K562 cells were cultured with different concentrations of HHT or in its combination with mTOR inhibitor rapamycin (RAPA) for 24 hours. The cell viability was analyzed by CCK-8 assay, the cell apoptosis was detected by flow cytometry, the expressions of BCL-6, Caspase-3 and mTOR signal pathway related proteins was assayed by Western blot, the expression of BCL-6 mRNA was determined by RT-PCR.</p><p><b>RESULTS</b>The HHT inhibited proliferation and induced apoptosis of K562 cells in a concentration-dependent manner(r=0.970). With the increasing of HHT concentration, the expression level mTOR signal pathway related proteins increased(r=0.908), while the mRNA and protein expression levels of BCL-6 decreased(r=-0.961, r=-0.981), as compared with the HHT alone, the combination of HHT with RAPA could down-regulate the expression of mTOR signal pathway related protein and caspase-3, and up-regulated expression of BCL-6.</p><p><b>CONCLUSION</b>HHT induces apoptosis of K562 cells by inhibiting BCL-6 expression through mTOR signal pathway.</p>

Effect of Homoharringtonine Combined with Imatinib on the K562/G01 Cells and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of homoharringtonine(HHT) combined with imatinib(IM) on proliferation and apoptosis of K562/G01 cells and its potential mechanism.</p><p><b>METHODS</b>K562/G01 cells were cultured with HHT and/or IM. CCK-8 assay was used to detect cell proliferation. Cell apoptosis and phosphorylated tyrosine levels were analyzed by flow cytometry. The expression levels of p210, PI3K, p-Akt and Akt protein were determined by Western blot.</p><p><b>RESULTS</b>Compared with HHT or IM alone, drug combination significantly inhibited cell proliferation and induced apoptosis of K562/G01 cells (both P< 0.05). HHT combined with IM could inhibit the levels of phosphorylated tyrosine and phosphorylated Crkl and downregulate the expressions of p210, PI3K and p-Akt in K562/G01 cells.</p><p><b>CONCLUSION</b>HHT combined with IM can synergistically inhibit proliferation and induce apoptosis of K562/G01 cells by suppressing the p210 expression and its kinase activity.</p>

Acadesine Inhibits the Proliferation of K562 Cells and Enhances their Sensitivity to Imatinib / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of AMPK agonist Acadesine (AICAR) on growth inhibition of K562 cells and their sensitivity to imatinib (IM).</p><p><b>METHODS</b>K562 cells were cultured with different concentrations of AICAR alone or its combination with IM for 48 hours, the CCK-8 assay was used to detect cell proliferation, the cell cycle distribution and apoptosis were analyzed by flow cytometry. The expression levels of Cyclin D1, Cyclin E1 and Caspase 3 protein were determined by Western blot.</p><p><b>RESULTS</b>AICAR inhibited the proliferation of K562 cells in dose-dependent manner, and their IC50 value was 0.45 mmol/L at 48 hours. AICAR could induce arrest of K562 cells in G1 phase and down-regulated the protein expression levels of Cyclin D1 and Cyclin E1; whereas it didn't influence the cell apoptosis. Additionally, the growth inhibition of cells induced by IM was enhanced by AICAR.</p><p><b>CONCLUSION</b>AICAR can inhibit the proliferation of K562 cells by arresting the cell cycle and enhancing the sensitivity of K562 cells to IM.</p>

Effects of Homoharringtonine Combined with Imatinib on K562 Cell Apoptosis and BCL6 expression / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effects of homoharringtonine (HHT) alone or combined with imatinib (IM) on K562 cell proliferation and apoptosis, as well as the mRNA and protein expression of BCL6.</p><p><b>METHODS</b>The CCK-8 was used to detect the inhibitory effect of drugs on cell growth, the flow cytometry was used to detect the cell apoptosis. The expression of BCL6 protein was assayed by Western blot, and BCL6 mRNA expression was detected by RT-PCR.</p><p><b>RESULTS</b>HHT alone displayed a proliferation inhibition effect with dose- dependent manner, and induced apotosis; after combination of HHT and IM drugs, both the inhibitory rate and the apoptosis rate were significantly increased compared with the drug alone(P<0.05). Western blot showed that the expression of BCL6 protein was down-regulated after being treated with HHT, however, the BCL6 protein was up-regulated after being treated with IM . The effect of drug combination showed that BCL6 protein significantly down-regulated(P<0.05 ). The expression of BCL6 mRNA was decreased both in the treatment of HHT or IM alone when compared with control. And the effect of drug combination showed that BCL6 mRNA expression was more significantly down-regulated(P<0.05 ).</p><p><b>CONCLUSION</b>HHT can inhibit the K562 cell proliferation and induce apoptosis of K562 cells. Combination of IM with HHT shows a significant synergistic effect, the mechanism may be associated with down-regulation of BCL6.</p>

Effect of YM155 on Apoptosis and Autophagy of K562 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was purposed to investigate the effect of YM155, a survivin inhibitor, on the apoptosis and autophagy of K562 cells.</p><p><b>METHODS</b>K562 cells were treated with YM155 at different concentration. Cell survival was analyzed by CCK-8 assay, the cell apoptosis was detected by flow cytometry. Survivin, BCL-2 and beclin1 mRNA expressions were determined by RT-PCR. Survivin, BCL-2, caspase-3, PARP and LC-3 protein expressions were assayed by Western blot.</p><p><b>RESULTS</b>YM155 inhibited the proliferation of K562 cells in a time- and dose-dependent manners. With the increasing of YM155 concentration and prolonging of action time, the expression levels of mRNA and protein of survivin and BCL-2 decreased, while the expression levels of caspase-3, PARP, beclin1 and LC-3 increased. Compared with the YM155 group, the protein levels of LC-3 and caspase-3 were lower in YM155 combined with 3-MA group.</p><p><b>CONCLUSION</b>YM155 can inhibit K562 cell proliferation by inducing apoptosis and autophagy, while autophagy induction effect can enhance its cytotoxic effect.</p>

Influence factors of salt-sensitive hypertension and responses of blood pressure and urinary sodium and potassium excretion to acute oral saline loading among essential hypertensive patients / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the influence factors of salt-sensitive hypertension and to observe changes of blood pressures and urinary sodium and potassium excretion in response to acute oral saline loading among essential hypertensive patients in China.</p><p><b>METHODS</b>Essential hypertensive patients from Beijing Jinzhan second community were included in this study. Salt-sensitivity was determined via the improved Sullivan's acute oral saline loading and furosemide volume-depletion tests. Binary logistic regression analysis was applied to explore influence factors of salt-sensitive hypertension. Acute oral saline loading induced changes on blood pressures and urinary sodium and potassium excretion were observed.</p><p><b>RESULTS</b>Sixty-three salt-sensitive hypertensive patients were classified out of a total of 342(18.4%) essential hypertensive patients. Salt-sensitive patients were elder than the non-salt-sensitive patients (P < 0.05) . Binary logistic regression analysis showed that age (OR = 1.744, 95%CI:0.922-3.300, P > 0.05) , gender (OR = 0.728, 95%CI:0.374-1.415, P > 0.05) , total cholesterol level (OR = 1.168, 95%CI:0.882-1.547, P > 0.05) and 24-hour urinary sodium (OR = 0.998, 95%CI:0.995-1.002, P > 0.05) were not influencing factors of salt-sensitivity among essential hypertensive patients. Bivariate general linear models for repeated measures showed that there were significant statistical differences on blood pressures and urinary electrolytes concentrations between the beginning of trials, 2 hours after acute saline loading and 2 hours after furosemide volume-depletion(all P < 0.01). There was a greater blood pressures change in salt-sensitive patients than in non-salt-sensitive patients(all P < 0.01) while urinary electrolytes concentrations change was similar between two groups(all P > 0.05).</p><p><b>CONCLUSIONS</b>Age, gender, total cholesterol level and 24-hour urinary sodium are not influencing factors of salt-sensitivity among essential hypertensive patients in this study. Impaired pressure natriuresis during acute oral saline loading and furosemide volume-depletion tests is presented in salt-sensitive essential hypertensive patients.</p>

Expression of M3 subtype of muscarinic receptor during the skin incised wound healing in mice / 法医学杂志

OBJECTIVE@#To investigate the expression of M3 subtype of muscarinic receptors (M3R) during the incised wound healing of the skin in mice and the characteristics of its time-dependent.@*METHODS@#The change of M3R in skin incised wound was detected by immunohistochemical staining and Western blot.@*RESULTS@#M3R-positive cells were detected in epidermis, hair follicle, sebaceous glands, sweat glands, dermomuscular layer in normal mouse skin. Expression of M3R was mainly detectable in polymorphonuclear cells (PMNs) in the wound specimens aged from 6h to 12h after injury. Afterwards, the M3R-positive cells were mostly mononuclear cells (MNCs) and fibroblastic cells (FBCs) at 1 d to 3d post-injury, whereas the M3R-positive cells were mostly FBCs aged from 5 d to 14d. Morphometrically, the ratio of the M3R-positive cells increased aged from 6h to 12h after injury, with a peak at 12h. The ratios kept a high relatively level aged from 1 d to 5 d, but significantly that lowered as compared with aged 12h after injury. The ratio reached the peak at 7 d again after injury, and then decreased gradually. The M3R protein also revealed a time-dependent tendency with double peaks at 12h and 7 d after injury as detected by Western blotting.@*CONCLUSION@#M3R is time-dependently expression in PMNs, MNCs and FBCs suggesting that it may play roles during the skin incised wound healing, and M3R may be used as a marker for wound age determination.