Knockout of BMAL1 Gene Induces Apoptosis of HL-60 Cells and Inhibits its Proliferation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the biological function of BMAL1 in human acute myeloid leukemia by means of the HL-60 cell line in whica circadian gene BMAL1 was konocked-out by the CRISPR/Cas9 technology.</p><p><b>METHODS</b>Two sgRNAs for BMAL1 were designed and the PX459 knockout vectors containing the sgRNA were constructed. The activity of 2 sgRNAs was detected by T7 endonuclease I. the BMAL1 knocked out HL-60 cells were prepared by transient transfection of the target vectors into the cells. Western blot was used to detect the expression of BMAL1 protein. The apoptosis of the targeted cells was detected by flow cytometry. The proliferation status of the cells was assessed by the CCK-8 assay.</p><p><b>RESULTS</b>The PX459-sgRNA vectors were successfully constructed and screened to assure the activity of the targeting vector. It was found that the expression of BMAL1 protein was not detected in BMAL1-knocked out HL- 60 cells. Further, it was shown that BMAL1 knockdout could promote the apoptosis of HL-60 cells and inhibit the cell proliferation ability.</p><p><b>CONCLUSION</b>BMAL1 knocked out HL-60 cells have bean successfully established using the CRISPR/Cas9 gene editing technique, and BMAL1 knockout can promote the HL-60 cell apoptosis and inhibit its proliferation.These result reveal the biological role of the BMAL1 circadian gene in acute myeloid leukemia.</p>

Optimization of Extraction Process for Walnut Quinone from Juglans Green Peel by Box-Behnken Response Surface / 中国中医药信息杂志

Objective To optimize extraction process of walnut quinone from Juglans green peel. Methods On the basis of single factor tests, taking material-liquid ratio, extracting temperature and ethanol concentration as independent variables, yield of walnut quinone in response to as a value, according to Box-Behnken experiment design principle, extraction process of walnut quinone from Juglans green peel was optimized by response surface analysis. Results The optimum extraction process of walnut quinone was: 14.30 times the amount of 89.81% ethanol;30.28 ℃ constant temperature. The extracting amount of walnut quinone was 2.034 mg/g, which was close to the experimental results of 1.957 mg/g. Conclusion The optimized extraction process is reasonable and feasible, which can provide reference for the extraction of walnut quinone from Juglans green peel.

Mechanisms of sorafenib induced NB4 cell apoptosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of sorafenib on human acute promyelocytic leukemia cell NB4 and its mechanism.</p><p><b>METHODS</b>The human acute promyelocytic leukemia cell NB4 was treated with different concentrations (0, 1.5, 3, 6 and 12 µmol/L) of sorafenib, the proliferation inhibitory rate of NB4 cells was assayed by MTT, the apoptosis of NB4 was determined with flow-cytomatry after treatment; after extraction of total protein, the Western blot was performed to determine the expressions of apoptosis-relatived molecules Caspase-3, Caspase-8 and MCL-1. The mRNA expressions of Caspase-3, Caspase-8 and MCL-1 were determined by RT-PCR.</p><p><b>RESULTS</b>As compared with the control group, the proliferation of NB4 significantly decreased after treatment with different concentrations of sorafenib. The sorafenib significantly induced the apopotosis of NB4 cells in time- and dose-dependent manners. Furthermore, sorafenib treatment resulted in the obvious increase of the Caspase-3 and Caspase-8 protein and mRNA expressions, and down-regulated the MCL-1 protein and mRNA expressions in NB4 cells.</p><p><b>CONCLUSION</b>Sorafenib can inhibit proliferation and induce apopotosis of human acute promyelocytic leukemia cell NB4 through the expression of Caspase-3 and Caspase-8, and down-regulation of the expression of MCL-1.</p>

Caspase-independent apoptosis induced by arsenic trioxide in human multiple myeloma cell RPMI8226 / 中国实验血液学杂志

This study was purposed to explore the caspase-independent apoptosis pathway in human multiple myeloma cell RPMI8226 induced by arsenic trioxide (As(2)O(3)). MTT method was used to analyze the proliferation inhibition rate; flow cytometry was used to detect the apoptosis rate; Western blot was used to determine the expressions of BCL-2 and Caspase-3 in RPMI8226 cells. The results showed that As(2)O(3) (0.1 - 20 µmol/L) significantly inhibited the proliferation of RPMI8226 (P < 0.05) in concentration- and time-dependent manner. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, the apoptosis rate of zVAD-fmk (20 µmol/L) and As(2)O(3) combined treated group did not change. Compared with the group treated with As(2)O(3) (10 µmol/L) alone, zVAD-fmk (20 µmol/L) combined with As(2)O(3) (10 µmol/L) treatment group showed significant increase of expressions of Caspase-3 and BCL-2. It is concluded that As(2)O(3) can inhibit the proliferation of RPMI8226 cells. As(2)O(3) can induce apoptosis of RPMI8226 cells, and a caspase-independent process probably exist in As2O3-inducing RPMI8266 cells apoptosis.

Photodynamic therapy of condyloma acuminata in pregnant women / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is an emerging technique for the treatment of genital human papillomavirus (HPV)-induced benign and premalignant lesions. We report here in a case series of condyloma acuminata (CA) in pregnancy successfully treated with ALA-PDT.</p><p><b>METHODS</b>Five pregnant patients with CA received three to four times treatment respectively. Patients were followed up for 6 - 23 months after treatment.</p><p><b>RESULTS</b>The clearance rate of genital warts was 100%. No recurrence was found during the follow-up period. Major adverse events reported were mild erosion, pain, and local edema. All pregnancies resulted in healthy live births without delivery complications.</p><p><b>CONCLUSIONS</b>PDT with topical ALA seems to be safe and effective in the treatment of CA in pregnancy. It demonstrated high clearance rate of warts, was well-tolerated by patients, and showed no adverse effects on mothers or fetuses. ALA-PDT may be an ideal strategy of treatment for pregnant women with CA.</p>

The clinical significance of the epithelial cellular adhesion molecule(Ep-CAM)expression in esophageal squamous cell carcinoma(SCC) / 肿瘤研究与临床

Objective To evaluate the clinical significance of epithelial cellular adhesion molecule (Ep-CAM)expression in esophageal squamous cell carcinoma(SCC).Methods The Ep-CAM expression was immunohistochemically investigated in 70 normal esophageal mucosas,SCCs and 72 lymph nodes.Results Ep-CAM expression was observed in 94.3% of the tumors,but no expression in the normal mucosa.The Ep- CAM expression was not significantly different between different tumor scales and tumors invading depths,its expression level was relevant with the tumors differentiation and lymph node metastases(P