Effects of Emodin Derivative on Cell Cycle, Apoptosis and NF-κB Pathway in Burkitt Lymphoma Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and mechanism of a novel emodin derivative YX-18 on Burkitt lymphoma (BL) cells.@*METHODS@#MTT assay was used to detect the effect of YX-18 on the proliferation of BL cell lines CA46 and Raji. Annexin V-PE/7-AAD double staining assay was used for detecting the effect of YX-18 on the apoptosis of CA46 and Raji cells. PI/RNase staining was used to test the effect of YX-18 on CA46 and Raji cell cycle. JC-1 method was used to measure the changes of mitochondrial membrane potential after YX-18 treatment, and DAPI staining was used to detect the morphology of apoptotic cells. Western blot was used to analyze the distribution changes of NF-κB pathway protein (P65, P-P65, IκB, P-IκB) in the cytoplasm and cell nucleus, and also the expression changes of cyclin-related protein P21, CDK2, P-CDK2, Cycling D1, Cycling E1, and the apoptosis-related protein Caspase-3, Caspase-8, Caspase-9 and the proliferation-related protein C-MYC, BCL-2 by YX-18. Real-time fluorescence-quantitative PCR was used to evaluate the effects of YX-18 on mRNA levels of C-MYC and Ki-67 genes in CA46 and Raji cells, and EBNA-1 and EBER genes of EBV in Raji (EBV@*RESULTS@#Novel Emodin derivative YX-18 could effectively inhibit the proliferation of BL cell lines CA46 and Raji, showing a time-dependent effect (24, 48 and 72 h: r@*CONCLUSION@#The novel emodin derivative YX-18 can significantly inhibit the proliferation of Burkitt lymphoma cells, and induce the cell apoptosis and cycle arrest. The inhibitory effect of YX-18 on the proliferation of Burkitt lymphoma cells may be related with the effect of Caspase apoptosis pathway, the proliferation and apoptosis-related molecules, such as C-MYC and Ki-67, and also to the inhibition of NF-κB pathway.

Effect of Zhizhuwan on Intestinal Conduction and PLC-γ1/PLC-γ2 Signaling Pathway of Slow Transit Constipation Model Mouse with Spleen Deficiency Syndrome / 中国实验方剂学杂志

Objective:To explore the effect of Zhizhuwan on intestinal conduction and expressions of Phospholipase C-γ1 (PLC-γ1)/Phospholipase C-γ2 (PLC-γ2) signaling pathway of slow transit constipation (STC) with spleen deficiency syndrome. Method:Special pathogen free (SPF) healthy mice were randomly divided into normal group and model making group. Folium Sennae gavage was used to induce the spleen deficiency status, and then diet and drinking water were controlled to establish the mice model of spleen deficiency constipation. After the modeling, the mice in modelling group were randomly divided into model group, Zhizhuwan group and mosapride group. Zhizhuwan group was given drug at the dose of 9.0 g·kg-1·d-1, mosapride group was given 2.5 mg·kg-1·d-1, model group and normal group were given the equal dose of distilled water for 7 consecutive days. At the end of the treatment, the length of Indian ink in the colon was used to calculate the intestinal propulsion rate of the mice. The D-xylose kit was used to determine the content of D-xylose in serum of mice. The hematoxylin eosin (HE) staining was used to observe the pathological changes of colon tissues in mice. The immunohistochemistry and Western blot were used to detect the expression levels of PLC-γ1 and PLC-γ2 proteins in colon tissues of mice. Real-time PCR was used to detect the mRNA expression levels of PLC-γ1 and PLC-γ2. Result:Compared with normal group, the intestinal propulsive rate and the serum D-xylose value in model group were significantly decreased (Pγ1 and PLC-γ2 in colon tissues were significantly decreased (PD-xylose in Zhizhuwan group and mosapride group were significantly increased (Pγ1 and PLC-γ2 were significantly increased (PConclusion:Zhizhuwan can promote the intestinal movement in slow transit constipation model mouse with spleen deficiency syndrome, and alleviate the symptoms of constipation in mice. The related mechanism may be related to the increase of the expressions of PLC-γ1 and PLC-γ2 in colon tissues of mice with spleen deficiency and constipation.

Inhibitory and Inducing Effects of Emodin Derivative E11 on Proliferation and Apoptosis of Multiple Myeloma Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To screen the most strong emodin derivative inhibiting the proliferation of multiple myeloma(MM) cells and to explore the inhibitory and inducing effects of emodin derivatives on proliferation and apoptosis of MM cell lines RPMI 8226 and U266.</p><p><b>METHODS</b>Sixteen emodin derivatives were designed and synthesized by using emodin as mother substance, then from which the emodin derivative E11 was screened for experiments. The MTT method and cell colony formation assay were used to observe the effect of E11 on proliferation of RPMI 8226 and U266, the fluorescent microscopy with DAFI staining was used to observed the morphological changes of MM cells treated with emodin dervative 11, the DNA fragmentation detection was used to detect the inducing apoptosis effect of E11 on RPMI 8226 and U266 cells treated with E11.</p><p><b>RESULTS</b>The MTT assay showed that after the RPMI 8226 cells were treated with 16 kinds of emodin derivatives for 48 hours, the 50% inhibition concentration（IC） of 14 emodin dervatives was between 0.83-34.68 µmol/L, except E10 and E15 because their IC could not be calculated. The IC of E11 for RPMI 8226 and U266 cells were 0.831±0.0453 µmol/L and 1.039±0.093 µmol/L, respectively. Cell colony formation assay showed that E11 could inhibit RPMI8226 and U266 cells' colony formation in dose-.and time- dependent manner (r=0.72). Cell apoptosis was observed in RPMI8226 and U266 cells by DAPI staining , and also by the detection of DNA fragmentation.</p><p><b>CONCLUSION</b>In the synthesis of 16 kinds of emodin derivatives, the inhibitory effect of E11 on prolife-ration of RPMI8226 cell was the strongest. E11 can remarkably inhibit proliferation and induce apoptosis of RPMI8226 and U266 cells.</p>

Efficacy of sequential administration of artesunate and artesunate-amodiaquine for the treatment of falciparum malaria in South Sudan / 新乡医学院学报

Objective To investigate the efficacy of sequential administration of artesunate injection and artesunateamodiaquine tablets for the treatment of falciparum malaria in South Sudan.Methods The clinical data of thirty-one patients with falciparum malaria in Level One Hospital of the Chinese Peacekeeping Infantry Battalion from April 2016 to July 2017 were analyzed retrospectively.All patients were administered artesunate injection (120 mg for the first time,60 mg 4 h later,then 60 mg once daily) by intramscular injection for 1-3 d.The patients took orally artesunate-amodiaquine tablets (2 tablets) at 24,48,72 h after the body temperature restored to normal (totally 6 tablets).The efficacy of this sequential treatment for falciparum malaria was observed.Results The total cure rate of the 31 falciparum malaria patients was 100.0％ (31/31);the cure rate of patients after 3 days treatment was 90.3％ (28/31) and the cure rate of patients after 3-6 days treatment was 9.7％ (3/31).Mild side effects,such as nausea,vomiting,anorexia,abdominal distension,diarrhea or dizziness were observed in 4 patients (12.9％) and all these symptoms disappeared after the end of treatment.Conclusion The sequential administration of artesunate injection and artesunate-amodiaquine tablets has optimal curative efficacy on falciparum malaria in South Sudan,and the tolerance is well.

Effect of A Novel Emodin Derivative on Chronic Myelogenous Leukemia K562 Cells and Imatinib-resistant K562/G01 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of a novel emodin derivative E19 on proliferation inhibition and apoptosis induction of human chronic myelogenous leukemia (CML) cell line K562 and imatinib-resistant CML cell line (K562/G01), and to clarify the involved mechanisms.</p><p><b>METHODS</b>MTT and colony formation test were used to detect the cell proliferation. Apoptotic induction effects were examined by DAPI staining method and DNA ladder assay. Western blot was performed to detect the changes of P210(Bcr-Abl) protein.</p><p><b>RESULTS</b>The emodin derivative E19 could efficiently inhibit proliferation and induce apoptosis in K562 and K562/G01 cells. IC50 of K562 cells and IC50 of K562/G01 cells were (1.20 ± 0.19) µmol/L and (1.22 ± 0.16) µmol/L, respectively. DNA fragmentation in K562 cells and K562/G01 cells confirmed that the E19 induced apoptosis in dose-dependent manner. Western blot showed that emodin derivative inhibited phosphorylation of P210 protein in K562 cells and K562/G01 cells and down-regulated the expression level of P210 in dose- and time-dependent manners.</p><p><b>CONCLUSION</b>The emodin derivative E19 can efficiently inhibit growth and induce apoptosis of K562 cells and K562/G01 cells, while the inhibition of phosphorylation of P210 protein and down-regulation of P210 protein expression may be involved in these processes.</p>

Effect of Emodin Derivative E11 on T Lymphocytic Leukemia Cell Line Molt-4 and Its Possible Mechanisms / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of a new emodin derivative E11 on proliferation and apoptosis of T lymphocytic leukemia cell line Molt-4 and its possible mechanisms.</p><p><b>METHODS</b>MTT method was used to plot cell growth curve. Colony culture assay was performed for studying the effect of emodin derivative E11 on colony-formation of Molt-4. The fluorescent microscopy with DAPI staining was used to examine the cell morphological changes after E11 treatment. DNA fragmentation method was used to detect the inducing effect of emodin derivative E11 on cell apoptosis. Western blot was used to determine the expressions of apoptosis-related proteins including procaspase-9, procaspase-3, PARP and PI3K/AKT, MAPK signalling pathway.</p><p><b>RESULTS</b>Emodin derivative E11 could strongly inhibit the growth of Molt-4 with the IC50 in 48 h at 1.381 ± 0.1552 µmol/L in dose-dependent manner. 0.1 µmol/L of E11 could inhibit cell colony formation. The typrical apopototic morphologic changes of Molt cells treated with E11 could be observed under fluorescence microscope with DAPI staining. DNA apoptotic ladder could be observed by DNA fragmentation.The expressions of procaspase -9, procaspase-3, PARP, p-MAPK, p-AKT, mTOR, p-mTOR, p-P70 and p-4BEP1 were down-regulated, while expressions of MAPK, AKT, 4EBP1 and P70 were not changed remarkably after Molt-4 were treated with E11 for 48 h.</p><p><b>CONCLUSION</b>E11 can remarkably inhibit the proliferation and induce the apoptosis of Molt-4 cells. The mechanism of apoptosis of Molt-4 cells may be related with the suppression of PI3K/AKT and MAPK signalling pathways.</p>

Synthesis of emodin derivatives and their inhibiting effects on proliferation of leukemia cell lines / 中国实验血液学杂志

The aim of this study was to explore the inhibitory effect of newly synthesised emodin derivatives on the proliferation of leukemia cell lines and to select the most effective one from these emodin derivatives for further research. Emodin derivatives were synthesized by modifying the structure of emodin. MTT method was used to detect the proliferative inhibition in leukemia cell lines treated with emodin derivatives. The results showed that the half inhibitory concentration (IC50) for K562 cells treated with emodin derivatives E10-19 for 48 h was 0.84 - 12.01 µmol/L. E19 displayed the best anti-proliferative activity, while E16 and E17 did not show effects on K562 cells. Emodin derivative E19 was chosen for treating U937, NB4, Molt-4 and CA-46 cells, their IC50 for 48 h were 0.85, 0.9, 0.76, 0.8 µmol/L respectively. The IC50 of E19 for LQ2 cells was 3.60 µmol/L, and the IC50 range of E19 for normal human peripheral blood mononuclear cells at 48 h was 4.01 - 4.78 µmol/L. It is concluded that emodin derivative E19 can strongly inhibit the growth of leukemia cells and its inhibiting effect on proliferation of leukemia cells has a certain specificity. The specific mechanism of E19 anti-leukemia effect should be further studied.

Application of a simple random sampling method on surveys at the community level / 中华流行病学杂志

Objective To study the feasibility of a simple random sampling on surveys at the community level and to evaluate the quality of samples under survey.Methods A simple random sample of households was taken,based on the electronic listings of community households from Gongshu and Xiacheng districts of Hangzhou city.One of the adults aged 18 to 64 years in the sampled households was identified with KISH method to finish a questionnaire survey.More than 500 people from the sample size was required in each district.Results Of 950 sampled households in Xiacheng district,511 (53.8%) finished the survey while 506 (36.7%) out of the 1380 sampled households in Gongshu district did.The proportions of non-response due to the following reasons as:none with eligible age in the households,relocation of the original household,mass relocation of the community,and errors in the household listings etc.were 38.3% and 43.5% respectively,in the two districts.Proportions attributed to non-response and refusal to response of sampled household or individual were 8.0% and 19.9% respectively.No statistical significant differences in age and gender were found between the surveyed samples and the population in the sampled households,or in gender between the populations in the sampled households and in Hangzhou city.However,the population in the sampled households showed a more aging population structure than the population in Hangzhou city.Conclusion In a geographically limited area,using a simple random sampling method to do the survey is feasible,based on the electronic listings of household.Enough time spent during the household visit guarantees the interviewers to get a representative sample of the sampling frame.There is an urgent need for the timeliness,completeness and accuracy of electronic household listings to be improved.

Effect of extracted liquid from qianlietongyu on the proliferation and apoptosis on prostatic smooth muscle cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effect of extracted liquid from Qianlietongyu on the proliferation or apoptosis on prostatic smooth muscle cells in vitro.</p><p><b>METHODS</b>After extracted liquid from Qianlietongyu treated the cultured prostatic smooth muscle cells, the anti proliferative and apoptotic indices were assessed by MTT assy and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) respectively.</p><p><b>RESULTS</b>There was a significant dose-effect relationship between the concentration of extracted liquid from Qianlietongyu and the antiproliferative index on prostatic smooth muscle cells in vitro (P < 0.01), but there was no markedly difference in the apoptosis index between the group of extracted liquid from Qianlietongyu and control group ( P > 0.05).</p><p><b>CONCLUSION</b>Extracted liquid from Qianlietongyu may show significant antiproliferative effect on prostatic smooth muscle cells in vitro, without inducing apoptosis.</p>

The study of the pro-nucleating activity of bacteria identified in cholesterol gallstones in model bile systems / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the relationship of bacteria identified in cholesterol gallstones and gallstone formation.</p><p><b>METHODS</b>Observe the bacteria activity in model bile and the influence of bacteria on the cholesterol nucleation time (NT).</p><p><b>RESULTS</b>(1) Model bile were suitable for the growth of E. coli, Pseudomonas aeruginosa, staphylococcus aureus, enterococcus faecalis, clostridium difficile and Clostridium. Propionibacterium acne grew weakly and the growth of Bacteroides fragilis was restrained in model bile. (2) Only pseudomonas aeruginosa and enTerococcus faecalis could ly shorten the cholesterol nucleation time. (3) With pseudomonas aeruginosa or enTerococcus faecalis added in model bile, the formation of cholesterol crystals presented a progressive course of evolution.</p><p><b>CONCLUSIONS</b>Pseudomonas aeruginosa and enterococcus faecalis, not propionibacterium acne, have pro-nucleating ability in model bile.</p>