[Characteristics of Bone Marrow Cell Morphology and Immunophenotype in Patients with NHL Leukemia and Evaluation of NHL Bone Marrow Invasion].

OBJECTIVE: To observe the evaluation value of characteristics of bone marrow cell morphology and immunophenotype in patients with non-Hodgkin lymphoma (NHL) leukemia on the bone marrow invasion of NHL. METHODS: The clinical data of 104 patients with NHL treated in the hospital from March 2016 to March 2021 were retrospectively analyzed, the characteristics of bone marrow smear morphology were recorded, and the analysis of bone marrow immunophenotype was performed, the evaluation value of bone marrow cell morphology and immunophenotype on NHL bone marrow invasion was analyzed. RESULTS: One hundred and four patients with NHL leukemia were found to have increased lymphoma cells by examination of bone marrow cell morphology, including 57 cases of B-cell type, 39 cases of T-cell type and 8 cases of NK/T cell type. The characteristics of bone marrow cell morphology were as follows: the cell body was large, irregular or round like in shape, the cytoplasm was much and mostly stained blue, a few cells could see a few granules, the nucleus was round or round like, some were twisted, the chromatin was thick and nucleolus was different, of which the T-cell type lymphoma cells had strong heteromorphism and more obvious nucleolus. B-cell type mainly expressed CD19, HLA-DR and CD20, and the positive rate was ≥70%. T-cell type mainly expressed CD7, HLA-DR and CD38, and the positive rate was ≥40%. NK/T cell types mainly expressed CD56 and CD161, and the positive rates was ≥50%. Compared the proportion of lymphoma cells between bone marrow smear and immunophenotype examination, there was no statistical significant difference (P>0.05). CONCLUSION: The characteristics of bone marrow cell morphology and immunophenotype have certain application value in the evaluation of bone marrow invasion in patients with NHL leukemia, both can complement each other and provide a feasible mean for the effective evaluation of bone marrow invasion in patients with NHL leukemia.

[Comparison of chemical composition between fresh and processed Bufonis Venenum by UPLC-TQ-MS].

Toad venom is the Bufo bufo gargarizans or B. melanostictus after the ears of the gland secretion, used in the treatment of various cancers in recent years. Research shows that the main anti-tumor components in bufadienolide. Bufadienolide have free type structure and conjunct type structure. To identify and clarify the difference between bufogenin and bufotoxin contained in Bufonis Venenum, which was from B. bufo gargarizans, an UPLC-TQ-MS method has been established. UPLC-TQ-MS method was used to identify and quantify the major bufadienolides in Bufonis Venenum. UPLC-TQ-MS assay with positive ion mode was performed on a Waters ACQUITY UPLC BEH C, (2.1 mm x 100 mm, 1.7 µm) with the mobile phase consisting of 0. 1% aqueous formic and acidacetonitrile in gradient elution at a flow rate of 0.4 mL · min⁻¹ and the column temperature was set at 35 °C. By comparing their retention time and high resolution mass data of Bufonis Venenum extracts, 37 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were free-type bufadienolides (bufogenin), 11 of them were conjugated bufadienolides. There were significant differences in the main composition between fresh and processed Bufonis Venenum. The study found that the chemical composition of toad venom through great changes after processing, conjunct type content is much less, free type content as well change.

[Anti-apoptotic effect of insulin in myocardial ischemia-reperfusion and its principal signaling pathway].

OBJECTIVE: To investigate the signaling pathway involved in the insulin-elicited anti-apoptotic effect during myocardial ischemia and reperfusion (MI/R) in vivo. METHODS: Male Sprague-Dawley rats were anesthetized and subjected to 30 min of myocardial ischemia followed by 4h-reperfusion. Rats were randomly treated with intravenous infusion of saline (vehicle, 4 ml.kg(-1).h(-1)), insulin (60 U/L), or insulin + wortmannin 5 min before reperfusion and continuing throughout the 4h-reperfusion period. Cardiac myocyte apoptosis was determined both qualitatively and quantitatively by DNA laddering and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) methods. Myocardial nitric oxide (NO) was measured by using NO-specific chemiluminescence detector. Activations of Akt and p38 mitogen-activated protein kinase (MAPK) were determined by kinase activity assays using corresponding kinase activity assay kits (Cell Signaling). RESULTS: In the vehicle-treated rats, MI/R caused significant cardiac myocyte apoptotic death. Treatment with insulin produced a significant anti-apoptotic effect as evidenced by a marked reduction of apoptotic index [(8.0 +/- 2.9)% vs. (19.3 +/- 4.6)% of vehicle, P < 0.01] and decreased formation of myocardial DNA fragmentation. In addition, insulin treatment produced 2.7-fold increase (P < 0.01) of myocardial Akt activity and 28% increase of myocardial NO production (P < 0.05), while p38 MAPK activity changed insignificantly as compared with that of vehicle (P > 0.05). Both insulin-induced Akt activation and anti-apoptotic effect could be abrogated by wortmannin, a phosphatidylinositol (PI) 3-kinase inhibitor. CONCLUSION: In vivo treatment with insulin at the initial of reperfusion significantly reduced postischemic apoptotic death via the PI3-kinase-Akt signaling pathway. Akt, but not p38 MAPK, activation plays a key role in the insulin-induced anti-apoptotic effect in MI/R.

Enhancement of glutathione cardioprotection by ascorbic acid in myocardial reperfusion injury.

The present experiment determined the effects of glutathione and ascorbic acid, the two most important hydrophilic antioxidants, on myocardial ischemia-reperfusion injury and evaluated their relative therapeutic values. Isolated rat hearts were subjected to ischemia (30 min) and reperfusion (120 min) and treated with ascorbic acid, glutathione monoethyl ester (GSHme), or their combination at the onset of reperfusion. Administration of 1 mM GSHme alone, but not 1 mM ascorbic acid alone, significantly attenuated postischemic injury (P < 0.05 versus vehicle). Most interestingly, coadministration of ascorbic acid with GSHme markedly enhanced the protective effects of GSHme (P < 0.01 versus vehicle). The protection exerted by the combination of GSHme and ascorbic acid at 1 mM each was significantly greater than that observed with 1 mM GSHme alone (P < 0.05). Moreover, treatment with GSHme alone or GSHme plus ascorbic acid markedly reduced myocardial nitrotyrosine levels, suggesting that these treatments attenuated myocardial peroxynitrite formation. These results demonstrated that 1) GSHme, but not ascorbic acid, exerted protective effects against ischemia-reperfusion injury; and 2) the protective effects of GSHme were further enhanced by coadministration with ascorbic acid, suggesting a synergistic effect between GSHme and ascorbic acid.