Effect of modified FC/ATG preconditioning on allogeneic hematopoietic stem cell transplantation for severe aplastic anemia / 中华器官移植杂志

Objective:To evaluate the efficacy and safety of modified FC/ATG pretreatment in the treatment of severe aplastic anemia(SAA).Methods:From June 2012 to June 2020, clinical data of 64 patients with severe aplastic anemia undergoing allogeneic hematopoietic stem cell transplantation with modified FC/ATG(Flu 30 mg·m -2·d -l, -5~-2 d、CTX 50 mg·kg -1·d -1~-2 d、ATG: 2.5 mg·kg -1·d -1, -5~-2 d) pretreatment were retrospectively analyzed.There were MSD-HSCT ( n=29) and Haplo-HSCT ( n=35). Results:One patient died of intracerebral hemorrhage before transplantation and the remainders were completely implanted.During a median follow-up period of 14.5(1-95) months, overall survival (OS) rate of 92.2%.It was significantly higher than OS rate of 67.2% in the treatment of SAA by foreign pretreatment regimens containing low-dose TBI.And pretreatment scheme containing FC+ BU/TBI had an OS of slightly >91.3% in the treatment of SAA.The 3-year OS rates were 85.7% and 93.5% in Haplo-HSCT and MSD-HSCT groups ( P=0.058). The OS rate of SAA Haplo-HSCT/MSD-HSCT group was similar to that of " Beijing Protocol" (BU/CY+ ATG) (89%, 91%). The viral infection rates of EB and CMV were significantly higher in haplo-HSCT group than those in MSD-HSCT group and inter-group difference was statistically significant ( P<0.05). However, univariate analysis showed that two groups had no effect on survival time ( P=0.403, P=0.132). Univariate analysis showed that survival time was significantly associated with the presence of Ⅲ-Ⅳ° aGVHD and the presence of severe complications ( P=0.007, P=0.001). Further multivariate analysis revealed that severe complication was an independent risk factor for survival ( P=0.003). Conclusions:The efficacy of improved FC/ATG pretreatment in the treatment of SAA in MSD-HSCT or Haplo-HSCT is higher than other domestic and international pretreatment schemes in OS rate, safety and effectiveness.Onset of severe complication and association with Ⅲ ~ Ⅳ ° aGVHD are the influencing factors for patient survival.The efficacy of Haplo-HSCT group is similar to that of MSD-HSCT group.It may be employed as an alternative donor for SAA patients without fully congruent donors.

Analysis of PKD2 gene variant and protein localization in a pedigree affected with polycystic kidney disease / 中华医学遗传学杂志

OBJECTIVE@#To detect the mutation site in a pedigree affected with autosomal dominant polycystic kidney disease (ADPKD) and verify its impact on the protein function.@*METHODS@#Peripheral blood samples were collected from the proband and his pedigree members for the extraction of genomic DNA. Mutational analysis was performed on the proband through whole-exome sequencing. Suspected variant was verified by Sanger sequencing. A series of molecular methods including PCR amplification, restriction enzyme digestion, ligation and transformation were also used to construct wild-type and mutant eukaryotic expression vectors of the PKD2 gene, which were transfected into HEK293T and HeLa cells for the observation of protein expression and cell localization.@*RESULTS@#The proband was found to harbor a c.2051dupA (p. Tyr684Ter) frame shift mutation of the PKD2 gene, which caused repeat of the 2051st nucleotide of its cDNA sequence and a truncated protein. Immunofluorescence experiment showed that the localization of the mutant protein within the cell was altered compared with the wild-type, which may be due to deletion of the C-terminus of the PKD2 gene.@*CONCLUSION@#The c.2051dupA (p. Tyr684Ter) mutation of the PKD2 gene probably underlay the pathogenesis of ADPKD in this pedigree.

Identification of a novel SOD1 variant in a Chinese patient with amyotrophic lateral sclerosis / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese patient with amyotrophic lateral sclerosis (ALS).@*METHODS@#Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA. Genetic variant was identified by whole exome sequencing. Candidate variant was verified by Sanger sequencing of his parents and healthy controls.@*RESULTS@#The patient was found to harbor a heterozygous c.420C>G (p.Asn140Lys) variant of the SOD1 gene. The same variant was not detected in his parents and 100 healthy controls. The variant has not been included in HGMD, dbSNP and other databases.@*CONCLUSION@#The c.420C>G variant of the SOD1 gene may underlie the ALS in this patient. Above finding has enriched the spectrum of SOD1 gene variants.

Analysis of gene variant in a Chinese pedigree with preaxial polydactyly / 中华医学遗传学杂志

OBJECTIVE@#To analyze the pathogenic variant of preaxial polydactyly in a Chinese Han pedigree and identify the cause of polydactyly.@*METHODS@#The peripheral blood DNA of the proband and her parents was extracted. The polydactyly-related genes were detected by trio whole exome sequencing, and the suspected pathogenic gene was screened out. Sanger sequencing was applied to other members of the pedigree.@*RESULTS@#The results of gene sequencing showed that the LMBR1 gene had a heterozygous variant of c.423+4909(IVS5)C>T in 6 patients of the pedigree. The same variant was not detected in family members with normal phenotype. Based on the ACMG guidelines, c.423+4909(IVS5)C>T of the LMBR1 gene was predicted to be pathogenic (PM1+PM2+PP1-S(PS)+PP4+PP5).@*CONCLUSION@#The heterozygous C>T variant at position 4909 of intron 5 of the LMBR1 gene probably underlies the disease in this pedigree.

Efficacy and safety analysis of VCD and VD regimens for treatment of newly diagnosed patients with multiple myeloma / 白血病·淋巴瘤

Objective@#To compare the clinical efficacy and safety of bortezomib, cyclophosphamide, dexamethasone (VCD) regimen and bortezomib dexamethasone (VD) regimen in the treatment of the patients with newly diagnosed multiple myeloma (NDMM).@*Methods@#The clinical data of 73 patients with NDMM in Shanxi Dayi Hospital from January 2013 to January 2016 were retrospectively analyzed. According to the chemotherapy regimen, the patients were divided into VCD group (41 cases) and VD group (32 cases). The efficacy and adverse reactions of the two groups were evaluated.@*Results@#The overall response rate of VCD group and VD group was 80.5% (33/41) and 78.1% (25/32) respectively, and the difference was not statistically significant (χ 2= 0.061, P= 0.804); and complete remission (CR) rate was 36.6% (15/41) and 15.6% (5/32) respectively, and the difference was statistically significant (χ 2= 3.970, P= 0.046); the median progression-free survival (PFS) was 27 months and 24 months respectively, and the median overall survival (OS) was 35 months and 33 months respectively, and the differences were not statistically significant (all P > 0.05). Most adverse reactions occurred in grade 1-2, in which peripheral polyneuropathy and thrombocytopenia were the most common. Peripheral neuritis was the most common finding among the adverse reactions of grade 3. The incidence of adverse reactions in both groups was not statistically significant (P > 0.05).@*Conclusions@#VCD and VD regimen could be recommended as a better induction therapy for NDMM patients. Compared with VD scheme, VCD scheme has a higher CR rate.

Effects of the concentrations of glutathione and the related mechanisms of arsenic trioxide (As2 O3) on Multidrug Resist-ance-associated Pr otein / 中国医师杂志

Objective To investigate the apoptosis-inducing effect , inhibiting MRP-1 and mRNA expression of arsenic trioxide (As2O3) and buthionine sulfoximine (BSO) on multidrug-resistant cell-K562A/DM cell .To compare the effect of As2O3 and the com-bined group .To determine the effect of intracellular GSH content on the arsenic effect .Methods To investigate the effect of the arse-nic group (0.5,2.0,5.0μmol/L)and/or BSO (100μmol/L) on multidrug-resistant cell -K562/ADM cell.To detect the change of the correlated index .⑴Intracellular GSH contents were measured using Glutathione Assay Kit by spectrophotometry .⑵MRP-1 expression were determined by flow cytometry .⑶MRP-1 mRNA expression were directed by semi-quantitative RT-PCR.Results After the GSH contents were degraded by the combination of clinic dose arsenic group (0.5,2μmol/L) and BSO(100μmol/L), in 48 hours and 72 hours, the effect of the combination group ( clinic dose arsenic group ) was obviously stronger than the clinic dose arsenic group and the high dose arsenic group .In 48 hours, the MRP-1 mRNA depressive effect of the combination group ( clinic dose arsenic group ) was obviously stronger than high dose arsenic group .In 72 hours, the MRP-1 depressive effect of the combination group ( clinic dose arsenic group ) was obviously stronger than high dose arsenic group .Conclusions The intracellular GSH contents closely correlated with the arsenic effect .The combination of clinic dose arsenic and BSO inhibit obviously MRP-1 expression and MRP-1 mRNA expres-sion in K562/ADM cell.

Effect of the content of glutathione for multidrug resistance-associated protein 1 / 肿瘤研究与临床

Objective To investigate the apoptosis-inducing effect,inhibiting multidrug resistanceassociated protein 1 (MRP-1) and mRNA expression of arsenic trioxide (As2O3) and buthionine sulfoximine (BSO) in multidrug-resistant cell K562/ADM.To compare the effect of As2O3 and the combined group.To determine the effect of intracellular glutathione (GSH) content on the arsenic effect.Methods The arsenic group (0.5 μmol/L,2.0 μmol/L,5.0 μmol/L) solo or combined BSO (100 μmol/L) applied in multidrugresistant cell K562/ADM.The cell proliferating activity was assessed with MTT assay.The cell apoptosis effect was detected by Annexin-V and propidium iodide (PI) staining.Intracellular GSH contents were measured using GSH Assay Kit by spectrophotometry.MRP1 expression was determined by flow cytometry.MRP1 mRNA expression were directed by semi-quantitative RT-PCR.Results With the GSH contents were degraded by the combination of clinic dose arsenic group (0.5 μmol/L,2 μmol/L) and BSO (100 μmol/L),the K562/ADM cell proliferating activity was obviously inhibited and the cell apoptosis-inducing effect was increased in 24 hours.In 72 hours,the rate of apoptosis with arsenic (0.5 μmol/L,2 μmol/L) were (8.32± 2.11) ％,(16.75±3.56) ％.After the GSH contents were degraded,the rates of apoptosis in the combination group (clinic dose arsenic group) were (82.15±9.28) ％ and (92.72±9.41) ％.The fluorescence intensity of MRP1 in 72 hours of the combination group (clinic dose arsenic group) was 8.20±0.92 and 10.40±2.33.The MRP1 attenuated effect of the combination group (clinic dose arsenic group) was obviously stronger than that of the high dose arsenic group (21.30±3.09).Conclusions The intracellular GSH contents closely correlate with the arsenic effect.The cell apoptosis-inducing effect of the combination of clinic dose arsenic and BSO on K562/ADM cell is obvious increased.The combination of clinic dose arsenic and BSO obviously inhibit MRP1 expression and MRP1 mRNA expression in K562/ADM cell.

Growth inhibition and apoptosis induction to imatinib-resistant cell line K562/G01 by Bortezomib / 白血病·淋巴瘤

Objective To explore the effect of proteasome inhibitor bortezomib (BOR) on proliferation inhibition and apoptosis in imatinib-resistant chronic myeloid leukemia cell line K562/G01. Methods MTT assay was used to observe the effect of growth inhibitory of cells; flow cytometry was used to detect cell cycle and apoptosis. Results K562/G01 cells was not sensitive to imatinib,1C50 values of imatinib to K562 and K562/G01 cells was (20.09±1.38) and (0.54±0.13) μmol/L,respectively; BOR could inhibit proliferation in K562/G01 cell,peaked at 48 h,and IC50 value of BOR to K562/G01 was (23.10±2.71) nmol/L. G2/M phase cell cycle arrest and significant apoptosis peak could be seen by flow cytometry with increased in the concentration and action time of BOR. Conclusion BOR can inhibit proliferation and induce apoptosis in imatinib-resistant K562/G01 cell,the mechanism may be related to cell cycle G2/M phase arrest.

Association of CD+4 CDHigh25 regulatory T cells and the negative control factor IL-10 with the development of acute graft-versus-host disease after allogeneic hematopoietic cell transplantation / 白血病·淋巴瘤

Objective To investigate the relationship between the CD+4 CDHigh25 regulatory T cells and cytokine IL-10 and acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods Flow cytometric was used to detect the percentage of CD+4 CDHigh25Foxp3High Treg cell and CD+4 CDHigh25 CDLow127 Treg cell in CD+4 T cells and at the same time ELISA was used to test the serum IL-10 levels in corresponding period. Results 13 patients have received hematopoietic function reconstruction. aGVHD group of CD+4 CDHigh25 CDLow127/CD+4 and CD+4 CDHigh25 Foxp3High/CD+4 ratio were significantly lower than non-aGVHD group, the difference was statistically significant (P ＜0.01); Ⅲ-Ⅳ degree of aGVHD subgroup was lower than Ⅰ - Ⅱ degree of aGVHD subgroup, but no statistical significance(P ＞0.05); the same as between-group CD+4 CDHigh25 CDLow127/CD+4 and the CD+4 CDHigh25 Foxp3High/CD+4 has no significant difference;aGVHD group of IL-10 concentration was significantly lower than non-GVHD group, the difference was statistically significant (P ＜0.01). Treg cell and IL-10 changes in correlation, r = 0.557, P ＜0.05. Conclusion The level of Treg cell was closely related to the occurrence of aGVHD after allo-HSCT. So it is very important to monitor the Treg cell level for clinical early diagnosis of aGVHD and predict prognosis of aGVHD and guide the application of immunosuppressant. CD127 can serve as a Treg cell surface-specific marker, to promote the detection of Treg cell and purification. IL-10 was an important negative regulator. Treg cell and IL-10expression in patients with aGVHD was correlation, which may provide some basis for Treg cell immunosuppressive mechanism.

Research advances of CD+4CD+25 Treg cell and it's function in GVHD / 肿瘤研究与临床

CD+4 CD+25 regulatory T cell (Treg) has immune incompetent and immune suppression functions, and is a kind of suppressor T-cell subsets. It can inhibit the activation and proliferation of CD+4 T cells and CD+8 T cells, so as to effectively suppress the immune system to foreign organ and reduce the graftversus-host disease (GVHD) after hematopoietic stem cell transplantation (HSCT), without affecting the role of graft-versus-leukemia (GVL), which plays an important role in the transplantation immune tolerance.