Effect of Serum Free Light Chain on Renal Function and Prognosis in Patients with Newly Diagnosed Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To analyze the effect of serum free light chain (sFLC) on renal function and prognosis in patients with newly diagnosed multiple myeloma (MM).@*METHODS@#The clinical data of 70 newly diagnosed MM patients who received sFLC examination in Fujian Medical University Union Hospital were retrospectively analyzed from April 2012 to November 2016. Univariate analysis was used to analyze the risk factors that associated with renal impairment (RI) and prognosis. Logistic regression and Kaplan-Meier analyze were used to analyze the roles of sFLC in RI and the prognosis.@*RESULTS@#Out of the 70 patients, 20 patients had RI at the initial diagnosis. Compared to normal renal function group, RI group had lower level of hemoglobin, elevated levels of serum uric acid, corrected calcium, serum creatinine, serum β2 microglobulin, and involved sFLC, higher proportion of patients with ISS stage III, involved sFLC≥500 mg/L, hemodialysis (all P＜0.05). Multivariate logistic regression analysis showed that serum uric acid≥430 μmol/L, ISS stage III and a involved sFLC≥500 mg/L were all the independent risk factors for RI in patients with newly diagnosed MM patients (all P＜0.05). Receiver operating characteristic (ROC) curves analysis showed that the involved sFLC was 705.0 mg/L, which was a best cut-off value area under curve (AUC) for prediting RI in patients with MM was 0.727 (P=0.003), sensitivity was 65.0% and specificity was 82.0%). After a median follow-up period of 31 (1-84) months, the median overall survival (OS) of patients with involved sFLC≥500mg/L and involved sFLC＜500 mg/L were 52.0 and 27.0 months, respectively, there was no statistically significant difference (P=0.137). There was also no statistically significant difference in median OS between the high sFLC ratio group (κ/λ＞32 or ＜0.03) and the low sFLC ratio group (0.03≤κ/λ≤32) (27 months vs 40 months, P=0.436).@*CONCLUSION@#The involved sFLC in the RI group is significantly higher than that in the normal renal function group in newly diagnosed MM patients. Serum uric acid≥430 μmol/L, ISS stage III and involved sFLC≥500 mg/L are the independent risk factors for RI. Monitoring sFLC in newly diagnosed MM patients is helpful to the prediction of RI, and the involved sFLC level or sFLC ratio may not affect the prognosis of newly diagnosed MM patients.

Homing characteristics of graft cells in leukemia mice after allogenic bone marrow transplantation with different conditioning regimens / 中国实验血液学杂志

The aim of this study was to investigate the homing characteristics of bone marrow cells in leukemia mice after allogenic bone marrow transplantation with different conditioning regimens on the basis of a leukemia mouse model. Allogenic bone marrow transplantation was performed after three different kinds of conditioning regimen, including nonmyeloablative conditioning regimen (5 Gy (60)Co γ ray total body irradiation, A group), radiotherapeutic myeloablative conditioning regimen (9 Gy (60)Co γ ray total body irradiation, B group) and chemotherapeutic myeloablative conditioning regimen (large dose chemotherapy, C group). In the recipient mice, the nucleated cell number in peripheral blood, bone marrow and spleen was counted, the percentage of positive cells capable of connecting with FITC labeled anti-mouse H-2K(b) antibody was detected by flow cytometry and the homing ratio in bone marrow and spleen was calculated at 24, 48, 72, 96 h after bone marrow transplantation. The results showed that donor myeloid cells displayed homing and then mobilization (going out of home) in group A; homing, mobilization, and rehoming in group B and C, and there was a little delay of homing in the spleen in group C. In bone marrow, the homing efficiency of A group was the highest in early period and the lowest [(0.90 ± 0.09)%] in the fourth day with the mobilization of myeloid cells (P < 0.05), and the homing efficiency of B and C groups was lower in the early period and the highest [(2.17 ± 0.26)%, B group] in the fourth day with the rehoming of myeloid cells (P < 0.05). In spleen, the homing efficiency was similar to that in bone marrow and there still was a little delay in C group. It is concluded that the homing ratio is high in the early period and decrease obviously in 72 h after bone marrow of leukemia mice treated with nonmyeloablative conditioning regimen. The homing ratio is low in the early period and increases obviously in 72 h after bone marrow of leukemia mice treated with radio-or chemotherapeutic myeloablative conditioning regimens. The homing ratio does not obviously change between the early period and 72 h after bone marrow of leukemia mice treated with chemotherapeutic myeloablative conditioning regimen, and lies between group A and B.

Apoptosis-inducing effect of C-MYC siRNA on acute lymphoblastic leukemia Jurkat cell line / 中国实验血液学杂志

The study was purposed to investigate the effects of C-MYC siRNA on the proliferation and apoptosis of acute lymphoblastic Jurkat cell line. siRNA targeting the site 1545-1565 of C-MYC mRNA was designed and chemically synthesized, then C-MYC siRNA was transfected into Jurkat cells by the transfer agent (HiPerFect Transfection Reagent), the morphological changes were observed under inverted microscope; the tetrazole compound (MTS) was applied to draw the cell growth curve; the cell colony test was used to detect the effect of C-MYC siRNA on the proliferation of Jurkat cells; the flow cytometry and TUNEL method were used to analyze the apoptosis of Jurkat cells. The results showed that after Jurkat cells were treated with different concentrations of C-MYC siRNA, the growth of Jurkat cells was inhibited to various degrees, inhibitory rate was enhanced as C-MYC siRNA concentration increased. C-MYC siRNA also could obviously inhibit the cell clony formation. The apoptosis of cells could be detected by flow cytometry and TUNEL method, the apoptosis rate of cells increased along with prolonging of treatment with C-MYC siRNA. It is concluded that the chemically synthesized C-MYC siRNA can inhibit significantly the proliferation and induce the apoptosis of Jurkat cells.

Effects of small interference RNA against c-myc on expression of c-myc and h-tert genes in acute lymphoblastic leukemia cell line Jurkat / 中国实验血液学杂志

This study was purposed to investigate the effect of small interfering RNA against c-myc on c-myc, h-tert gene and protein expressions in acute lymphoblastic leukemia cell line (Jurkat cells), so as to provide new methods and targets for gene therapy of leukemia. The siRNA against target sites 1545-1565 of c-myc mRNA was chemically synthesized and was transfected into Jurkat cells by transfectant. The c-myc, h-tert mRNA and protein expression levels before and after treatment with c-myc siRNA were detected by RT-PCR and Western blot, respectively. The results showed that c-myc siRNA obviously inhibited the proliferation of Jurkat cells, the half inhibitory concentration (IC50) for 48 hours was about 75 nmol/L. c-myc siRNA could decrease c-myc, h-tert mRNA and C-MYC, h-TERT protein expression levels of Jurkat cells. It is concluded that c-myc siRNA significantly reduce c-myc, h-tert mRNA and protein expression levels through inhibiting c-myc mRNA expression and decreasing intracellular level of C-MYC protein.

The effect of inhibiting EOLA1 expression in ECV304 cells / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the effect of inhibiting the expression of endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) on proliferation of human umbilical vein endothelial cell line ECV304.</p><p><b>METHODS</b>After constructing and transfecting EGFP-EOLA1 fusion protein expressive vector into ECV304 cells, the transfected cells was cultured in M199 containing G418 for 5 weeks to screen the cell line stable expression EGFP-EOLA1 fusion protein. Oligonucleotides targeting EOLA1 at different sites were synthesized and inserted into pSinencer3.1/H1 vector. Then, the recombinant vector was transfected into the cultured ECV304 cells and the inhibiting effect to target gene EOLA1 was investigated by observing the green fluorescence in transfected cells under inverted fluorescent microscope and by Western blot assay. The proliferation of ECV304 cells was numbered when the expression of EOLA1 in ECV304 cells was inhibited by RNA interference.</p><p><b>RESULTS</b>The ECV304 cell line stably expressing EGFP-EOLA1 fusion protein was constructed and the siEOLA1 interfere vectors can knock down EOLA1 gene expression specially. When blocking the expression of EOLA1 in ECV304 cells,the proliferation of cells slowed down.</p><p><b>CONCLUSION</b>EOLA1 maybe has a role on the proliferation of cells.</p>

Thermal stress can inhibit proliferation of ECV304 cells / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To observe the effects of thermal stress on proliferation of human vascular endothelial cells (VECs) and explore its significance.</p><p><b>METHODS</b>Changes of VECs proliferation were investigated with (3)H-TdR incorporation method after ECV304 was treated at 43 degrees for 2 hours, while expressions of intercellular adhesion molecule-1 (ICAM-1), inhibitor of differentiation-1 (ID1), and P16 and P21 proteins were determined by Western Blotting.</p><p><b>RESULTS</b>The effect of inhibition of VECs growth after thermal stress was detected by (3)H-TdR incorporation experiment. Western blotting showed ICAM-1, a marker of activated endothelial cells, was increased markedly after thermal stress. Expression of ID1 protein declined gradually with increasing expressions of its downstream genes, P16 and P21 following the thermal stress.</p><p><b>CONCLUSIONS</b>Thermal stress could strongly activate VECs and inhibit proliferation of VECs through ID1, thus down regulating cyclin-dependent kinase inhibitors, P16 and P21, which might be an essential pathway for recovery of VECs after thermal stress.</p>