Effect of Chemotherapeutic Drug-Induced Damage of Bone Marrow Stroma Cells on Normal Hematopoietic Cells / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of damage of bone marrow stroma cells induced by chemotherapeutic drug on the function of normal hematopoitic cells.@*METHODS@#Senescence cells were detected by flow cytometry after SA-β-gal staining; real-time PCR was used to detect the expression of a serial molecules in bone marrow stromal cell line OP9 cells; the expression of γ-H2AX was determined by flow cytometry after histone γ-H2AX staining; the colony forming ability of hematopoietic cells was tested by colony formation assay.@*RESULTS@#The percentage of senescence cells in OP9 cells after DNR treatment was 2.24 times as much as that in untreated OP9 cells (P<0.05). Compared with normal OP9 cells, the expression levels of IL-6 and TNF-alpha in DNR-treated OP9 cells increased by 2.73 times (P<0.01) and 0.56 times (P<0.01), and the expression levels of N-cadherin, alpha smooth muscle actin (alpha-SMA), angiopoietin1 (Angpt1) and osteopontin (OPN) decreased by 69.54%（P<0.01），63.90%（P<0.01），87.41%（P<0.01）and 42.78%（P<0.01）respectively. After the co-culture with DNR-treated OP9 cells, the colony formation of normal hematopoietic cells decreased by 47.10% than that co-cultured with untreated OP9 cells (P< 0.05), meanwhile, the percentage of γ-H2AX+ cells in normal hematopoietic cells increased by 2.19 times (P<0.05).@*CONCLUSION@#After treatment with DNR, the senescence cell number of OP9 cells sgnificantly increases; the expression of TNF-α and IL-6 is up-regulated, while the expression of α-SMA, Angpt-1 and OPN is down-regulated as compared with normal OP9 cells. In addition, after co-culture of DNR-treated OP9 cells with normal hematopoietic cells, the colony formation ability of hematopoietic cells decreases and the genome instability of hematopoietic cells increases as compared with normal hematopoietic cells.

Oxidative Damage of Bone Marrow Stromal Cells Caused by Chemotherapy Drugs / 中国实验血液学杂志

OBJECTIVE@#To explore the oxidative damage of OP9 cells induced by daunorubicin (DNR) treatment.@*METHODS@#The TMRM probe was used to detect mitochondrial membrane potential by flow cytometry; the reactive oxygen species (ROS) was determined by flow cytometry DCFDA probe; the real-time PCR was used to detect the molecular expression of antioxidant enzyme，glutathione peroxidase (GPX) in OP9 cells; the expression of γ-H2AX was determined by flow cytometry.@*RESULTS@#Compared with normal OP9 cells, the positive rate of TMRM in DNR-treated OP9 cells decreased by 56.7% (P＜0.05); the positive rate of DCFDA in DNR-treated OP9 cells increased by 3.52 times (P＜0.01). Compared with normal OP9 cells, DNR-treated OP9 cells showed a decrease in the expression of GPX4 by 44.22% (P＜0.001); the expression of GPX7 decreased by 65.7% (P＜0.001); the expression of GPX8 decreased by 24.7% (P＜0.001); the positive rate of γ-H2AX in DNR-treated OP9 cells increased (P＜0.05).@*CONCLUSION@#After DNR treatment, mitochondrial membrane potential of OP9 cells decreases; the level of reactive oxygen species increases; the expression of glutathione peroxidase (GPX) molecules decreases significantly; genomic instability increases obviously; the oxidative damage of cells increased.

Expression and Asymmetric Division of Numb in Leukemia Cell K562 Line / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of asymmetric division in leukemia cells through detection of expression and asymmetric division of Numb in differentiated and undifferentiated K562 cells.</p><p><b>METHODS</b>Firstly, Hemin was used to induce K562 cell differentiation, and the expression of Numb was detected by the real-time quantitative RT-PCR and flow cytometry. After K562 cells were synchronized by nocodazole, the Numb protein was labeled by immunohistochemical staining, followed by the determination of the terminally differentiated cells through confocal microscopy. The fluorescence intensity was calculated by Image J software, and the cell division pattern was analyzed on the basis of the fluorescence intensities of Numb in 2 divided daughter cells.</p><p><b>RESULTS</b>Compared with the undifferentiated K562 cells, the level of Numb mRNA expression increased 2.3 times (P<0.001). The ratio of Numb positive cells was(67.37±5.01)% in differentiated K562 cells, while that was (43.97±5.72)% in undifferentiated K562 cells (P<0.01). Compared with undifferentiated K562 cells, the ratio of cells with asymmetric division in differentiated K562 cells increased 18.3%, the percentage of cells with symmetry self-renewal reduced 49.7%(P<0.001) and that with symmetry differentiation increased 32%(P<0.001).</p><p><b>CONCLUSION</b>In differentiated K562 cells, expression of Numb and proportion of cells with asymmetric division were higher than that in undifferentiated cells. With the differentiation of leukemia cells, the proportion of cells with asymmetrical division increases, and the proportion of cells with symmetrical self-renewal decreases. The stemness of leukemia cells is maintained mainly through the symmetrical self-renewal.</p>

Preliminary study of the association between human thrombospondin and gastric cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the possible role of human thrombospondin (hPWTSR) in gastric cancer and explore its potential to serve as the target for gastric cancer diagnosis and intervention.</p><p><b>METHODS</b>Using pLexA-hPWTSR as the bait, a premade pB42AD-based fetal brain cDNA library was constructed to identify the interacting proteins. The expression pattern of hPWTSR in gastric cancer tissues and a gastric cancer cell line was observed to investigate the correlation between hPWTSR expression and the biological behaviors of the tumor. The possibility of hPWTSR as a potential gastric cancer marker was evaluated.</p><p><b>RESULTS</b>Fifty-seven independent clones were isolated from 107 clones screened. Sequence analysis indicated that the 57 positive clones represented the products of 12 genes. A RT-PCR-based expression pattern revealed that the expression of hPWTSR in gastric cancer tissues and a gastric cancer cell line was lower than that in the corresponding normal tissues, but no mutations were identified by the subsequent sequence analysis.</p><p><b>CONCLUSIONS</b>hPWTSR interacts with adhesion-related proteins and tumor-related genes, and its expression is lowered in gastric cancer tissues and gastric cancer cell line. hPWTSR might play a role in gastric cancer development, especially in metastasis and might be used as a potential gastric cancer marker. The exact functions of hPWTSR and its potential clinical value still await further study.</p>

Study of the effect of the low dosage endotoxin damage on the expression of the organic cation transporter (OCT1) mRNA in hepatocytes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To elucidate the effect of the low dosage endotoxin damage on the expression of the organic cation transporter 1 (OCT1) mRNA in hepatocytes and make an approach to the probable effect of dexamethasone on the expression of the OCT1 mRNA after endotoxin damage.</p><p><b>METHODS</b>(1) The endotoxin damage model was established in rats; (2) The change of the expression of OCT1 mRNA in hepatocytes after endotoxin damage was observed by in situ hybridization method; (3) The change of the ultra structure of hepatocytes after endotoxin damage was observed with the electron microscope; (4) Dexamethasone was injected intraperitoneally before endotoxin damage in order to determine the influence of dexamethasone on the expression of OCT1 mRNA after endotoxin treatment.</p><p><b>RESULTS</b>The expression of OCT1 mRNA decreased until 16 hours (0.5745+/-0.012, P<0.01) after endotoxin treatment and then increased after this time point, which was still lower than the normal control; The expression of OCT1 mRNA in rat hepatocytes increased at each time point after endotoxin damage with dexamethasone treatment. It was highest at 16 hours (0.6327+/-0.007, P<0.01) after endotoxin damage, but it was still lower than that of the normal control.</p><p><b>CONCLUSION</b>Endotoxin could repress the expression of OCT1 mRNA even before the low dosage endotoxin inducing serious damage to the structure of hepatocytes; Dexamethasone could not induce the expression of OCT1 mRNA in normal hepatocytes, but could lighten the repression of endotoxin on the expression of OCT1 mRNA. And then the expression of OCT1 mRNA could increase at some extent after endotoxin damage with dexamethasone treatment.</p>

RCT on the therapautical effect of mospride citrate in functional dyspe psia / 第三军医大学学报

Objective　To observe the curative effect and sid e effect of the gastrokinetic agent mosapride citrate by RCT. Methods　 42 cases of functional dyspepsia (FD) were divided into two groups rando mly, the group of mosapride(21 cases):orally administrated mosapride, 5mg, t.i.d for 4 weeks, and the control (21 cases):orally administrated domperidone, 5mg, t.i.d for 4 weeks. Symptoms and side effects were recorded before and at d 14, d 28 after administration of the medicines according to GCP and double blind pri ciple. Gastric empting test was also carried out in randomly selected patients. Results　Mosapride and domperidone were significantly effective on alleviating symptoms of the patient with FD. In mosapride treated group the half emptying time was shortened and the 120 min remain rate was reduced. No sid e effect was found. Conclusion　These results suggest that mosa pride 5 mg t.i.d. is effective and safe on alleviating symptoms of patients with FD and improving the ga stric empting time.

RCT on the therapautical effect of mospride citrate in functional dyspe psia / 第三军医大学学报

Objective　To observe the curative effect and sid e effect of the gastrokinetic agent mosapride citrate by RCT. Methods　 42 cases of functional dyspepsia (FD) were divided into two groups rando mly, the group of mosapride(21 cases):orally administrated mosapride, 5mg, t.i.d for 4 weeks, and the control (21 cases):orally administrated domperidone, 5mg, t.i.d for 4 weeks. Symptoms and side effects were recorded before and at d 14, d 28 after administration of the medicines according to GCP and double blind pri ciple. Gastric empting test was also carried out in randomly selected patients. Results　Mosapride and domperidone were significantly effective on alleviating symptoms of the patient with FD. In mosapride treated group the half emptying time was shortened and the 120 min remain rate was reduced. No sid e effect was found. Conclusion　These results suggest that mosa pride 5 mg t.i.d. is effective and safe on alleviating symptoms of patients with FD and improving the ga stric empting time.