Objective To study the effect of metformin on the growth of megakaryocytic leukemia cell line Dami and to explore the molecular mechanisms of the inhibitory effect of metformin on the proliferation of Dami. Methods The Dami cells were cultured and divided into control and 1,2,4,8,16 and 32 mmol·L-1 metformin groups.Then MTT test was performed to detect the inhitory rate of proliferation of Dami cells after treated with different concentrations of metformin. Flow cytometry was used to examine the distribution of cell cycle, and Western blotting was carried out to analyze the expressions of Cdc2 and CylinB1 and the phosphorylation of Cdc2. Results The MTT results showed that compared with control group,the inhibitory rates of proliferation of the Dami cells in 32 mmol·L-1 metformin groups at 0,24,48,72 and 96 h (35.1%±2.3%,49.7%±5.1%, 78.85±0.9%,79.1%± 3.0%%,and 85.2%± 3.2%)were significantly increased(P<0.01),Furthermore, after metformin treatment for 72 h,the inhibitory rates of proliferation of the Dami cells in 1,2,4,8,16 and 32 mmol·L-1 metformin groups were (33.8 ± 0.3)%,(51.9 ± 0.2)%,(59.4 ± 1.6)%,(65.5 ± 2.0)%, (75.5±0.9)%,and (79.1±3.0)%,respectively. Metformin inhibited the growth of Dami cells in a time-and dose-dependent manner. The flow cytometry results results revealed that compared with control group, the percentages of Dami cells in G2/M phase in 1,2 and 4 mmol·L-1 metformin groups were increased from (26.0± 0.5)% to (38.5 ± 1.5 )%, (48.4 ± 1.1 )%, and (58.2 ± 2.7 )%;there was significant difference in the percentages of Dami cells in G2/M phase between control group and 4 mmol·L-1 metformin group (P<0.01). Western blotting analysis showed that compared with control group, the expressions of Cdc2 and CyclinB were evidently reduced, the phosophorylation of Cdc2 at Tyr1 5 was up-regulated, and the phosphorylation at Thr1 6 1 was down-regulated.Conclusion Metformin can inhibit the growth of Dami cells and induce G2/M arrest,and its mechanism may be related to inhibiting the activation of Cdc2/CyclinB1 complex.

OBJECTIVETo detect the role of surviving (SVV) in the protective effect of resveratrol against hypoxia/reperfusion injury (H/RI) of cardiac microvascular endothelial cells (CMECs).

METHODSCMECs isolated from the hearts of adult rats were exposed to hypoxia (94% N₂, 5% CO₂, 1% O₂) for 2 h followed by 4 h reoxygenation (95% O₂, 5% CO₂). The cell proliferation of CMECs was measured by MTT assay and Transwell method was used to detect migration ability of CMEC, PI-AnnexinV double staining and flow cytometry technique were employed to observe the apoptotic rate of CMECs. The SVV protein expression was detected with Western blot method.

RESULTSCompared to control group, the proliferation (0.19 ± 0.03 vs. 0.42 ± 0.07, P < 0.01) and migration ((28 ± 2)/5HPF vs. (50 ± 3)/5 HPF, P < 0.01) abilities were impaired and the apoptosis index ((19.7 ± 0.8)% vs. (5.4 ± 0.3)%, (P < 0.05) of CMEC was increased after H/RI. The proliferation (0.36 ± 0.07 vs. 0.19 ± 0.03, P < 0.05) and migration ((55 ± 3)/5HPF vs. (28 ± 2)/5HPF, P < 0.05) abilities of CMEC were significantly improved while the apoptosis index ((9.6 ± 0.7)% vs. (19.7 ± 0.8)%, P < 0.05) was significantly decreased in H/RI+resveratrol group compared to H/RI group.SVV protein expression was also upregulated in H/RI+resveratrol group compared to H/RI group (P < 0.05). To further ascertain the role of SVV in the protective effects of resveratrol, PI3K specific inhibitor LY294002 was added to H/RI+resveratrol group, the proliferation (0.25 ± 0.05 vs. 0.36 ± 0.07, P < 0.05) and migration ((34 ± 3)/5HPF vs. (55 ± 3)/5HPF, P < 0.05) abilities were significantly decreased, the apoptosis index ((16.2 ± 0.6)% vs. (9.6 ± 0.7)%, P < 0.05) was increased and the protein expression of SVV was downregulated (P < 0.05) in LY294002+H/RI+resveratrol group compared to H/RI+resveratrol group.

CONCLUSIONResveratrol could significantly reduce H/RI induced apoptosis and attenuate H/RI induced cardiac microvascular endothelial cells dysfunction through up-regulating PI3K/Akt/SVV pathways.

Animals ; Apoptosis ; Cell Proliferation ; Chromones ; Endothelial Cells ; Enzyme Inhibitors ; pharmacology ; Heart ; Hypoxia ; Morpholines ; Myocardium ; Myocytes, Cardiac ; Phosphatidylinositol 3-Kinases ; drug effects ; metabolism ; Proto-Oncogene Proteins c-akt ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; Stilbenes ; pharmacology ; Up-Regulation

Sialidosis is a rare lysosomal storage disease caused by NEU1 gene mutation at 6p21.33. It is characterized by myoclonic, ataxia, epilepsy, and decreased vision. A pair of twins with sialidosis type 1 are reported to enrich clinicians ′ understanding of the disease, so as to improve the diagnosis and treatment. The proband was a 16-year-old male. The main symptom was intermittent limb involuntary trembling for 2 years, with paroxysmal loss of consciousness. Fundus examination showed cherry-red spots. His twin brother had similar symptoms, but the overall performance was mild. Whole exome sequencing results showed that both patients carried compound heterozygous mutations of c.239C>T (p.P80L) and c.803A>G (p.Y268C) in NEU1 gene, which were from their normal phenotype mother and father.

Objective:To investigate the prognostic value of colony forming unit-granulocyte and macrophage (CFU-GM) in allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:Seventy-three patients who received allo-HSCT in Hebei Yanda Lu Daopei Hospital from February 2015 to January 2017 were selected. According to the level of CFU-GM from bone marrow (BM) culture at the time of allo-HSCT, the patients were fit into high CFU-GM group and low CFU-GM group. The overall survival rate (OS) and relapse-free mortality rate (NRM) of patients after transplantation were tested by χ2 test after a follow-up of 37.0 (12.5, 50.5) months. Kaplan-Meier method was used to compare OS and event-free survival (EFS) of patients with different CFU-GM levels. Logistic regression model was used to analyze the prognostic factors. Cox regression model was used to further analyze the prognostic risk of patients.Results:Compared with the low CFU-GM group, the high CFU-GM group had a higher OS (81.40% vs 60.00%, χ2=4.067, P=0.044) and a lower NRM (11.63% vs 36.67%, χ2=6.474, P=0.011). Compared with the low CFU-GM group, the mean OS time (57.6 and 37.1 months, respectively, P=0.039) and the mean EFS time (61.7 and 38.5 months, respectively, P=0.011) were significantly higher in the high CFU-GM group. Logistic regression analysis showed that both the level of CFU-GM and BM MNC were significant influencing factors of OS ( OR=2.917, 95% CI 1.011-8.418, P=0.048 and OR=1.510, 95% CI 1.058-2.154, P=0.023, respectively) and EFS ( OR=4.400, 95% CI 1.336-14.492, P=0.015 and OR=1.447, 95% CI 1.002-2.090, P=0.049, respectively)after transplantation. The level of CFU-GM was an independent risk factor for evaluating EFS ( HR=0.279, 95% CI 0.097-0.805, P=0.018). BM MNC was an independent risk factor for OS ( HR=1.345, 95% CI 1.052-1.720, P=0.018). Conclusion:The level of CFU-GM and BM MNC were related to the prognosis of allo-HSCT. The patients in the high CFU-GM group had higher EFS.

Marinesco-Sj?gren syndrome (MSS), also known as hereditary ataxia-dwarf-mental retardation syndrome, is a rare autosomal recessive ataxia syndrome. This article reviews the recent advance in clinic characteristics, pathogenic gene mutation sites, pathogenesis and clinic diagnosis and treatment of MSS, in order to improve clinicians' understanding of the disease and diagnosis and treatment level, and reduce the missed diagnosis and misdiagnosis of the disease.

Objective:To investigate the relationship between the levels of serum cytokines and chemokines and the prognosis of patients with acute B-ALL after receiving chimeric antigen receptor (CAR)-T cell immunotherapy and acute graft-versus-host disease (aGVHD) in patients after bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:According to the case-control principle, Forty-two patients with B-ALL who received CD19-CAR-T cell immunotherapy bridged to allo-HSCT at Heibei Yanda Ludaopei Hospital from September 18, 2019 to May 9, 2022 were enrolled. Mann-Whitney U test was used to compare the changes of aGVHD-related cytokines and chemokine levels between CAR-T cell immunotherapy and bridging transplantation in different patients at the same time. Their plasma levels of cytokines and chemokines related to aGVHD were monitored at the day before CAR-T therapy and after CAR-T treatment at day 4, 7,14,21,28. The receiver operating characteristic curve was drawn to evaluate the predictive value of cytokines and chemokines in predicting the occurrence and the death of aGVHD patients. Kaplan-Meier method and Log-rank tests were used for Overall survival (OS) analysis. Results:Twenty-four of total 42 patients had aGVHD, of which 11 patients died and 31 patients survived. There was no significant difference in cytokines and chemokines between the aGVHD group and the non-aGVHD group on the day before CAR-T cell treatment. According to statistical analysis, the serum Elafin levels of aGVHD group was higher than that of non-aGVHD group at the 21st day [4 482 (2 811, 6 061) ng/L vs 2 466 (1 948, 3 375) ng/L, Z=3.145, P=0.001] and the 28st day [4 391 (2 808, 5594) ng/L vs 2 463 (1 658, 2 830) ng/L, Z=2.038, P=0.048] separately. At the 14th day, serum cytokines and chemokines levels between the two group were as follows,MIP-1 α [21.02 (12.36, 30.35) ng/L vs 5.56 (3.64, 10.79) ng/L], sCD25 [422.47 (257.99, 1 233.78) IU/ml vs 216.11 (133.75,457.39) IU/ml], Elafin [4 101 (2 393, 5 006) ng/L vs 2 155 (1 781, 3 033) ng/L], IL-6 [119.08 (23.97, 183.43) ng/L vs 8.39 (2.91, 17.42) ng/L] and IL-8 [13.56 (12.50, 24.52) ng/L vs 2.83 (1.73,6.87) ng/L] were at higher levels ( Z=2.653, P=0.007; Z=2.176, P=0. 030; Z=2.058, P=0.041; Z=3.329, P<0.001; Z=3.162, P=0.001). The KM survival curve showed that the cumulative survival rates of patients with higher serum levels of MIP-1α, sCD25, Elafin, IL-6 and IL-8 were lower than those with low levels at day 14, and the difference was statistically significant (χ 2=12.353, 4.890, 6.551, 10.563, 20.755, P<0.05). Conclusion:The outcomes of patients treated with CAR-T cell therapy bridged to allo-HSCT was correlated with serum MIP-1α, sCD25, Elafin, IL-6 and IL-8 levels after receiving CAR-T therapy. High concentrations of MIP-1α, sCD25, Elafin, IL-6 and IL-8 suggest poor prognosis and can be used as biomarkers to suggest appropriate clinical selection of therapy.