The effect of 5-fluorouracil on enriching cancer stem cells of hepatoma cell line BEL-7402 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of 5-FU (5-fluorouracil) on enriching cancer stem cells of HCC cell line BEL-7402 and the biological characteristics of enriched cells.</p><p><b>METHODS</b>The enriching concentration of 5-FU was determined by CCK-8 (cell counting kit-8). Flow Cytometry was used to determine the changes in cell cycle and positive expression ratio of surface marker CD56, CD54, EpCAM and CD133. The self-renewal and differentiation of positive cells were tested by colony formation assay, and were compared with the control group.</p><p><b>RESULTS</b>Enriching concentration of 5-FU was determined as 10 μg/ml with 48 h incubation. After enrichment, G0/G1 phase cells increased from 57.50 %+/-0.98% to 68.70%+/-3.41% (P<0.05). Whereas S phase cells decreased from 40.26%+/-4.12% to 31.80%+/-4.15% (P<0.01); G2/M phase cells disappeared in experimental group, and was 5.80%+/-1.87% in control group (P<0.01). The proportion of the cell cycle changed with significant statistical differences. Meanwhile, positive rate of cell surface makers CD56, CD54, EpCAM and CD133 increased from 0.57%+/-0.12%, 8.10%+/-6.79%, 0.3%+/-0.01% and 3.20%+/-0.99% to 4.13%+/-0.06%, 50.08%+/-1.69%, 0.55%+/-0.07% and 10.51%+/-1.13%, respectively. The difference was significant (P<0.05). The colony forming ratio of CD56, CD54, EpCAM and CD133 negative cells and positive cells were 2.11%+/-0.21%, 3.32%+/-0.31%; 0.86%+/-0.101%, 2.40%+/-0.52 %; 7.19%+/-0.56%, 7.73%+/-0.71%; 2.70%+/-0.26%, 5.75%+/-0.81%, respectively, and significant differences were found between (P<0.05).</p><p><b>CONCLUSION</b>5-fluorouracil enriched the cancer stem cell population in HCC cell line BEL-7402. CD56 and CD54 can be used as important surface markers in research of liver cancer stem cells.</p>

Changes and mechanism of apoptosis-related gene expression in T lymphocytic leukemia JM cells induced with matrine / 中国实验血液学杂志

This study was purposed to investigate the changes of apoptosis-related gene expression in T lymphocytic leukemia JM cells induced with matrine, and its possible mechanism. JM cells was induced with 0.4 mg/ml matrine for 4 days, the total RNA was extracted from JM cells before and after matrine induction, the differential expression of apoptosis-related genes were screened with cDNA Expression Array Kit, the expression change of a part of gene was checked by Western blot. The results indicated that after induction of JM cells with matrine, differential expression of 31 genes were found by gene chip hybridization, the expression of caspase 8 was up-regulated more than 5 times. Western blot analysis showed that the up-regulation of caspase 8 gene expression positively correlated with induction time. It is concluded that differential expressions of many apoptosis-related genes in JM cells can be induced by matrine, in which gene expression of caspase 8 is up-regulated notably.

Study on the preparation of resveratrol chitosan nanoparticles with free amino groups on the surface / 中国中药杂志

<p><b>OBJECTIVE</b>To prepare resveratrol chitosan nanoparticles with free amine groups on the surface so as to conjugate ligands, which will actively target to special tissues or organs.</p><p><b>METHOD</b>The chitosan nanoparticles with free amine on the surface was prepared by sodium chloride precipitation. Nanoparticles with different solidification degrees were studied on turbidity, in vitro release, encapsulation efficiency, drug loading and diameter.</p><p><b>RESULT</b>The turbidity of nanoparticles with various solidification degrees decreased at different rates after ultrasonic or water bath heating treatment. All nanoparticles mentioned above obviously shew sustained release. The rate of release was slowed down with the increase of solidification agents. Solidification had no obvious effects on the encapsulation efficiency and drug loading. The diameter of chitosan nanoparticles with 200 microL solidification agents was 487 nm. The polydispersion was 0.144.</p><p><b>CONCLUSION</b>The diameter of the prepared nanoparticles was relatively small. The amine on the surface was free, which offered the possibility of designing the acive target drug delivery system.</p>

Expression of COX-2,CX-43 and P-gp protein in recurrent primary epithelial ovarian cancer / 肿瘤研究与临床

normal ovarian group (P

Inclusion bodies of human cytomegalovirus are composed of the DNA and immediately early and early antigens of the virus / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the composition and significance of the inclusion bodies of human cytomegalovirus (HCMV).</p><p><b>METHODS</b>Microdissection of inclusion bodies, PCR and Southern blot were adopted to detect DNA, and immunohistochemistry method and catalyzed signal amplification (CSA) were used to detect the different antigens of HCMV.</p><p><b>RESULTS</b>The inclusion bodies of HCMV were separated from the tissue section of human salivary gland. The fragments amplified by PCR from these dissected inclusion bodies were confirmed to be the DNA of HCMV. With the immunohistochemical method CSA, the immediately early and early antigens of HCMV were detected with monoclonal antibodies DDG9/CCH2, while matrix protein AAC10 was negative in the inclusion bodies.</p><p><b>CONCLUSION</b>The ingredient of inclusion bodies of HCMV included the DNA and the antigens expressed in specific stage of the virus.</p>

Effects of human fibroblastoid stromal cell line on proliferation of HL-60 cells and expression of VEGF / 中国实验血液学杂志

To investigate the effects of normal human bone m arrow fibroblastoid stromal cell line (HFCL) on the proliferation of acute myeloid leukemia cell line HL-60 and expression of vascular endothelial growth factor (VEGF), establishing coculture system of leukemia cell line HL-60 and HFCL, growth data was obtained by cell counting. Mitotic index (MI) was observed under Wright-Giemsa staining. Flow cytometry and Western blot were used as assays for cell cycle and expression of proliferating cell nuclear antigen (PCNA) separately. VE GF levels were evaluated by using commercial ELISA kits. The results showed that compared with HL-60 cells without HFCL cells, the proliferation of HL-60 cells in direct contact with HFCL cells and with HFCL cells separated by transwell was inhibited. The MI of HL-60 cells without HFCL cells was highest followed by HL-60 cells separated by transwell and HL-60 cells in direct contact with HFCL cells. The expression of PCNA in HL-60 cells with HFCL cells were lower than HL-60 cells without HFCL cells. Meanwhile, the percentage of HL-60 cells in G1 phase cocultured with HFCL cells was higher than that without HFCL cells while the percentage of Sphase cells was lower. The levels of VEGF in HL-60 cells with HFCL cells were lower than that in HL-60 cells alone. In conclusion, the normal bone marrow fibroblastoid stromal cells inhibited the proliferation of HL-60 cells as well as the expression of VEGF.

Matrine effects on JM cells by inhibiting proliferation and inducing apoptosis / 中国中药杂志

<p><b>OBJECTIVE</b>To study effects of matrine on JM cell strain.</p><p><b>METHOD</b>Morphologic changes were observed under light microscope with Wright-Giemsa staining, fluorescence microscope with Hoechst 33,258 staining and electron microscope. Alteration of cell cycle of different dose treating groups at the fourth day and 0.8 mg.mL-1 treatment group at the first, second, third, fourth day was analyzed by Flow cytometry. DNA ladder was detected with gel electrophoresis.</p><p><b>RESULT</b>From the third day after treatment of matrine, typical apoptosis features of cells were observed under light microscope and electron microscope in all test groups, and the features were more prominent with the time prolonging. At fourth day, flow cytometry analysis showed that there were sub-G1 peaks in all groups. From 0.1, 0.2, 0.4, 0.6 to 0.8 g.L-1 treatment groups, the rate of apoptotic cells to total cells were 3.1%, 2. 5%, 13.3%, 40.4%, 48.6%, respectively, and what in the control group was 1.4%; the rate of S phase cells to total cells was 28.9%, 26.1%, 27.7%, 0.9%, 14.2%, what in the control group was 30.4%; the rate of G1 phase cells to total cells was 63. 2%, 67.5%, 68.1%, 75.2%, 83.6%, what in the control group was 41.8%; From the first, second, third to fourth day, the rate of apoptotic cells to total cells of 0.8 mg.mL-1 treatment group were 3.0%, 3.7%, 9.1%, 48.6%, respectively; the rate of S phase cells to total cells was 28.6%, 17.5%, 19.1%, 14.2%; the rate of G1 phase cells to total cells were 45.5%, 77.3%, 77.2%, 83.6%. Gel electrophoresis displayed "DNA ladder" in 0.4, 0.6, 0.8 g.L-1 groups, while 0.1 and 0.2 g.L-1 groups didn't show such result.</p><p><b>CONCLUSION</b>Matrine can repress DNA synthesis and arrest JM cell strain at G1 phase, sequentially inhibiting the proliferation of the cell. Besides, this alkaloid can induce the apoptosis of JM cells.</p>