Role of Immune Microenvironmental Factors for Improving the IPI-related Risk Stratification of Aggressive B Cell Lymphoma / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the risk stratification of aggressive B cell lymphoma using the immune microenvironment and clinical factors.</p><p><b>METHODS</b>A total of 127 patients with aggressive B cell lymphoma between 2014 and 2015 were enrolled in this study. CD4, Foxp3, CD8, CD68, CD163, PD-1, and PD-L1 expression levels were evaluated in paraffin-embedded lymphoma tissues to identify their roles in the risk stratification. Eleven factors were identified for further evaluation using analysis of variance, chi-square, and multinomial logistic regression analysis.</p><p><b>RESULTS</b>Significant differences in 11 factors (age, Ann Arbor stage, B symptom, ECOG performance status, infiltrating CD8+ T cells, PD-L1 expression, absolute blood monocyte count, serum lactate dehydrogenase, serum iron, serum albumin, and serum β2-microglobulin) were observed among patient groups stratified by at least two risk stratification methods [International Prognostic Index (IPI), revised IPI, and NCCN-IPI models] (P < 0.05). Concordance rates were high (81.4%-100.0%) when these factors were used for the risk stratification. No difference in the risk stratification results was observed with or without the Ann Arbor stage data.</p><p><b>CONCLUSION</b>We developed a convenient and inexpensive tool for use in risk stratification of aggressive B cell lymphomas, although further studies on the role of immune microenvironmental factors are needed.</p>

Valproic Acid Enhances the Anti-tumor Effect of (-)-gossypol to Burkitt Lymphoma Namalwa Cells / 生物医学与环境科学（英文）

Burkitt lymphoma is a highly aggressive B-cell neoplasm. New therapeutic methods are needed to overcome the adverse effect of intensive chemotherapy regimens. Valproic acid and (-)-gossypol are two kinds of chemical compounds used as new anti-tumor drugs in recent years. To investigate the anti-tumor effect of valproic acid and (-)-gossypol, Burkitt lymphoma Namalwa cells were cultured and treated with valproic acid and (-)-gossypol at different concentrations. The proliferation of Namalwa cells was dramatically suppressed after the combination treatment with 2 mmol/L valproic acid and 5 μmol/L (-)-gossypol. The combined treatment also enhanced intrinsic apoptosis by down-regulating anti-apoptotic protein Mcl-1. Moreover, the autophagy flux significantly increased in Namalwa cells after combined treatment. However, the enhanced autophagy showed little effect on cell survival with present regimen. The results confirmed that combination of valproic acid and (-)-gossypol had synergistic anti-tumor effect to Burkitt lymphoma Namalwa cells. The related mechanisms might include the down-regulation of anti-apoptotic protein Mcl-1 and avianized pro-survival role of autophagy.

Exogenous gene expression in vitro and in vivo in bone marrow mesenchymal stem cells modified by hPDGF-A and hBD(2) / 中国实验血液学杂志

The objective of this study was to investigate the expression of exogenous hPDGF-A and hBD(2) in gene-modified bone marrow mesenchymal stem cells (BM-MSCs) in vitro and in vivo. Recombinant adenovirus vector expressing hPDGF-A/hBD(2) genes was constructed and packaged into virion. Primary isolated and cultured BM-MSCs were transfected by using hPDGF-A hBD(2), then the expressions of exogenous hPDGF-A/hBD(2) were detected by immunocytochemical staining in vitro. The conditioned medium (serum-free cultured supernatant of BM-MSCs transfected with recombinant adenovirus) collected from gene-modified BM-MSCs was applied to scratch wound on monolayer cells of multipotential cell line 10T1/2 in order to confirm the stimulative effect of hPDGF-A on cell migration. Gene-modified BM-MSCs were topically transplanted on wound of rats with radiation and skin excision combined injury. The distribution of BM-MSCs and expression of hPDGF-A/hBD(2) on the wound was observed by fluorescent microscopy and immunohistochemical staining respectively. The results indicated that the rat BM-MSCs transfected with recombinant adenovirus could express the EGFP in vitro. The immunofluorescent cytochemistry assay showed that the gene-modified BM-MSCs expressed the hPDGF-A and hBD(2). The scratch test confirmed that the percentage of healing area of wound in cultured supernatant group of gene-modified BM-MSCs was significant higher than that in control group on 8, 12, 24 and 48 hours (p < 0.05). The fluorescence microscopy of exogenous gene-modified BM-MSCs transplanted on wound revealed that the gene-modified BM-MSCs could higher express exogenous genes of EGFP at least within 2 weeks. The immunohistochemistry staining of wound indicated that the expression of exogenous genes began from day 3, reached to peak on day 7, and still visible on day 21 even though the expression became weak because of the possible dilution of the exogenous genes during cell division. It is concluded that efficient expression of exogenous hPDGF-A/hBD(2) in gene-modified BM-MSCs are demonstrated both in vitro and in vivo, which suggests that the molecular mechanism underlying chronic wound-healing accelerated by the strategy combining cell therapy with gene therapy.

The effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice, and to explore the possible underlying mechanism.</p><p><b>METHODS</b>Murine bone marrow cell (BMC) strain was prepared routinely and was employed in the establishment of the culture system of colony forming units of erythrocytes, or granulocytes and monocytes. To both sets of culture system normal murine serum (N group) and burn serum, which was collected from the mice with 15% full thickness burn at 12 postburn hours (PBH) and 1, 3, 5, 7 and 10 postburn days (PBD), (burn serum group) was added. In addition, positive control and blank control groups were set accordingly. The stimulating activity of all kinds of sera on the BMCs in the two sets of culture system was determined. The changes in the burn serum concentrations of EPO and GM-CSF were detected by radioimmunoassay, and the data were analyzed by logarithmic linear fitting correlation with the former influence of burn sera on the erythrocytes and granulocytes.</p><p><b>RESULTS</b>(1) Burn sera exhibited obvious stimulation promoting activity on the erythropoiesis and granulopoiesis in BMC, and the activity peaked (384 +/- 60 and 127 +/- 16 CFU) on 1 PBD and decreased thereafter to approach the values found in normal sera group (125 +/- 14 and 34 +/- 20 CFU) on 7 PBD. (2) The EPO content in burn serum was evidently higher than the normal value (P < 0.01) during 12 PBH to 7PBD period. The GM-CSF concentration was obviously higher than the normal value (P < 0.05) at 12 PBH and on 1 PBD. (3) The EPO concentration in burn serum was significantly and logarithmically correlated with the stimulation promoting activity of burn serum on erythropoiesis (r = 0.8570, P = 0.0137). But the GM-CSF concentration in culture with burn serum was not correlated with the stimulation promoting activity of burn serum on granulopoiesis (r = 0.7049, P > 0.05).</p><p><b>CONCLUSION</b>The sera harvested from burned mice during early postburn stage exhibited strong stimulation promoting activity on the erythropoiesis and granulopoiesis in bone marrow. The increased EPO level in burn serum might be the important factor contributing strong stimulation action on erythropoiesis, while increased GM-CSF level was not.</p>

Experimental study on preventive and therapeutic effec ts of 5 radioprotectants on combined radiation-burn injured mice / 第三军医大学学报

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

Effect of nordihydroguaiaretic acid on expressions of VEGF and its receptor KDR in vitro / 第三军医大学学报

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.

Experimental study on preventive and therapeutic effec ts of 5 radioprotectants on combined radiation-burn injured mice / 第三军医大学学报

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

Effect of nordihydroguaiaretic acid on expressions of VEGF and its receptor KDR in vitro / 第三军医大学学报

Objective　To explore the effect of nordihydroguaiaretic acid (NDGA) on the expressions of vascular endothelial growth factor (VEGF) and its receptor, kinase-inserted domain containing receptor(KDR) and the possible mechanism. Methods　The expression of VEGF in human malignant glioma cell line SHG-44 and that of KDR in human umbilical vein endothelial cell (HUVEC) line ECV-304 were observed 1～3 d after NDGA treatment with immunohistochemistry, in situ hybridization and image analysis. Results　The expression of VEGF was declined at protein or mRNA levels in SHG-44 cells after treated with 100 μmol/L NDGA for 1 to 3 d. The expression of KDR in endothelial cells with 100 μmol/L NDGA treatment for 1 to 3 d was decreased too, in a more obvious way compared with the decline of VEGF expression in SHG-44 cells. Conclusion　The results suggest that NDGA inhibits the expression of VEGF in glioma cells as well as that of VEGF receptor KDR in endothelial cells, which may be the important molecular mechanism of anti-angiogenesis of NDGA.