A telomere-related gene risk model for predicting prognosis and treatment response in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a prevalent hematological malignancy among adults. Recent studies suggest that the length of telomeres could significantly affect both the risk of developing AML and the overall survival (OS). Despite the limited focus on the prognostic value of telomere-related genes (TRGs) in AML, our study aims at addressing this gap by compiling a list of TRGs from TelNet, as well as collecting clinical information and TRGs expression data through the Gene Expression Omnibus (GEO) database. The GSE37642 dataset, sourced from GEO and based on the GPL96 platform, was divided into training and validation sets at a 6:4 ratio. Additionally, the GSE71014 dataset (based on the GPL10558 platform), GSE12417 dataset (based on the GPL96 and GPL570 platforms), and another portion of the GSE37642 dataset (based on the GPL570 platform) were designated as external testing sets. Univariate Cox regression analysis identified 96 TRGs significantly associated with OS. Subsequent Lasso-Cox stepwise regression analysis pinpointed eight TRGs (MCPH1, SLC25A6, STK19, PSAT1, KCTD15, DNMT3B, PSMD5, and TAF2) exhibiting robust predictive potential for patient survival. Both univariate and multivariate survival analyses unveiled TRG risk scores and age as independent prognostic variables. To refine the accuracy of survival prognosis, we developed both a nomogram integrating clinical parameters and a predictive risk score model based on TRGs. In subsequent investigations, associations were emphasized not solely regarding the TRG risk score and immune infiltration patterns but also concerning the response to immune-checkpoint inhibitor (ICI) therapy. In summary, the establishment of a telomere-associated genetic risk model offers a valuable tool for prognosticating AML outcomes, thereby facilitating informed treatment decisions.

Risk factors, outcomes and genotypes of carbapenem-nonsusceptible Klebsiella pneumoniae bloodstream infection: a three-year retrospective study in a large tertiary hospital in Northern China.

OBJECTIVE: To investigate the independent risk factors, outcomes and genotypes associated with carbapenem-non-susceptible K. pneumoniae bloodstream infections (BSIs) in northern China from 2014 to 2016. METHODS: Over a three-year period, a total of 289 K. pneumoniae BSI patients were identified. Medical records were extracted to obtain the clinical information. Polymerase chain reactions (PCRs) were performed to analyse the multilocus sequence typing (MLST) types, Klebsiella pneumoniae carbapenemase (KPC) and metallo-ß-lactamases (MBL) genes, for replicon typing of the 10 randomly selected carbapenem-non-susceptible K. pneumoniae. RESULTS: A total of 59 carbapenem-non-susceptible K. pneumoniae strains were identified. Resistance rates to imipenem, meropenem, ertapenem and amikacin were low. Multivariate analyses showed that a central venous catheter odds ratio (OR) of 4.021 (CI 1.002-16.134); mechanical ventilation of 7.587 (2.856-20.156); Pitt bacteraemia score of 1.481 (CI 1.218-1.800); hospitalization prior to culture of 1.026 (CI 1.001-1.053); and some antibiotic use 30 days prior to K. pneumoniae bacteremia, including carbapenem of 9.123 (CI 2.995-27.791), aminoglycoside of 34.079 (2.091-555.396), and tigecycline of 5.065 (CI 1.261-20.339) were associated with carbapenem-non-susceptible K. pneumoniae bacteremia. Sequence type 11 (ST11) was the most predominant MLST type, which accounted for 50% of the isolates. Eighty per cent of the isolates harbored the KPC-2 gene. The overall 28-day mortality rates of carbapenem-non-susceptible and carbapenem-susceptible K. pneumoniae were 54.24% and 19.56%, respectively. CONCLUSION: Central venous catheter, mechanical ventilation, high Pitt bacteraemia score, hospitalization prior to culture, and prior antibiotic use (carbapenem, aminoglycoside and tigecycline) were identified as independent risk factors for carbapenem-non-susceptible K. pneumoniae BSI, which was mostly caused by KPC-2 in northern China.

[An analysis of leukocyte subsets chimerism following allogeneic peripheral blood stem cell transplantation].

OBJECTIVE: To analyse the relationship of T lymphocyte and granulocyte chimerism following allogeneic peripheral blood cell transplantation and the occurrence of relapse, graft failure and graft versus host disease. METHODS: 21 patients underwent allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Fluorescence-activated cell sorter (FACS) sorted CD3+ T lymphocytes and CD15+ granulocyte from peripheral blood of all the patients were analyzed for short tandem repeats in 7 days interval for 1 month starting from the day of PBSCT, then 1 month interval for 6 months, and then 3 months interval to the end of one year. RESULTS: Chimerism of granulocyte was higher than T lymphocyte on day 7 posttransplant in 4 patients given myeloablative conditioning. The median donor chimerism of granulocyte and T lymphocyte was 95% and 55% respectively. The other 17 patients had higher chimerism of T lymphocyte than granulocyte on day 7, which was 60% (15%-76%) and 0% (0%-40%) respectively. 20 patients reached complete donor chimerism (CDC) on day 21 (14-102 days) for T lymphocyte and on day 14 for granulocyte, except one relapsed on day 28. Seven patients had decreasing mixed chimerism when disease relapsed or graft failure occurred. T cell donor chimerism decreased earlier than myeloid cells, however, bone marrow sample and granulocyte still remained in complete donor chimerism or stable mixed chimerism, bone marrow smear showed normal at the same time. CONCLUSION: Blood leukocyte subset chimerism analysis, especially T cell chimerism analysis may provide earlier information of engraftment, relapse and graft failure than blood and bone marrow samples, therefore immunomodulatory therapies may be given to recipients earlier and overall survival may be improved.

Is there a role for B lymphocyte chimerism in the monitoring of B-acute lymphoblastic leukemia patients receiving allogeneic stem cell transplantation?

OBJECTIVE: To determine the sensitivity and significance of B-cell chimerism for the detection of early engraftment, transplant rejection, and disease relapse. METHODS: The dynamic monitoring of lineage-specific cell subtypes (B, T, and NK cells) was made in 20 B-cell acute lymphoblastic leukemia (B-ALL) patients following allogeneic hematopoietic stem cell transplantation (allo-HSCT). In the early period after allo-HSCT, the latest establishment of B-cell complete chimerism (CC) was observed in a majority of patients. RESULTS: The percentage of donor cells of B-cell lineage was lower than the percent of T-cell lineage in most of the mixed chimerism (MC) patients. During graft rejection, the frequency of patients with decreasing MC of B-, T- and NK-cell lineage were 5/5, 2/5, and 2/5. When disease relapsed, five patients showed a faster decrease of the donor percent of B-cells than of T- or NK-cells. Only one patient displayed a more rapid decrease in NK-cells than in T- or B-cells. CONCLUSION: Monitoring of B-cell chimerism after HSCT seems to be valuable for insuring complete engraftment, anticipating graft rejection, and relapse in B-ALL patients.

Chimerism status is correlated to acute graft-versus-host disease after allogeneic stem cell transplantation.

To evaluate the correlation between chimerism status and acute graft-versus-host disease (aGVHD) following allogeneic hematopoietic stem cell transplantation. Chimerism of peripheral blood of 124 patients was monitored at regular intervals post-transplant. The chimerism of 124 post-transplant cases of CD3(+)T lymphocytes, 107 cases of CD3(-)CD56(+)CD16(+)NK lymphocytes, 49 cases of CD15(+) granulocytes, and 27 cases of CD19(+)B lymphocytes sorted by fluorescence-activated cell sorter were analyzed by polymerase chain reaction amplification of short tandem repeats. Differences were found in the time between establishment of full donor T-cell chimerism and the occurrence of aGVHD (P = 0.035, two related samples test). Patients with ≥69 % donor chimerism on day +7 in T-cells had higher rates of aGVHD. This study may provide a rational basis for treatment with adoptive immunotherapy at an earlier time, such as day 7 after SCT, than at present to prevent aGVHD.

[Item function analysis on the Quality of Life-Alzheimer's Disease(QOL-AD)Chinese version, based on the Item Response Theory(IRT)].

OBJECTIVE: To introduce the Item Function Analysis(IFA) of Quality of Life- Alzheimer's disease(QOL-AD)Chinese version and to explore the feasibility of its application on Chinese patients with AD. METHODS: Two hundred AD patients were interviewed and assessed by QOL-AD, through the stratified cluster sampling method. Multilog 7.03. was used for Item Function Analysis. Difference scale(a), difficulty scale(b)and Item Characteristic Curve(ICC) of each item of QOL-AD were provided. RESULTS: Different scales of the item 1, 7 were below 0.6, while all the others were above 0.6. As for ICC. The first and last lines for the other items were monotonic in which the two in between were in inverted V-shape, with very steep slopes, except for the item 1 and 7. CONCLUSION: Results form the IFA showed that QOL-AD was applicable to be used in the Chinese patients with AD.

[Comparison of STR-PCR and FISH value for monitoring chimerism after sex-mismatched allogeneic hematopoietic stem cell transplantation].

This study was purposed to compare the significance of multiplex short tandem repeat polymerase chain reaction (STR-PCR) and fluorescent in situ hybridization (FISH) for monitoring chimerism after sex-mismatched allogeneic hematopoietic stem cell transplantation (allo-HSCT). The chimerism of bone marrow or peripheral blood cells from 38 patients was analyzed by STR-PCR and FISH on days 14, 28 and at 3 months after allo-HSCT. The results indicated that on day 14, the complete chimerism (CC) was detected in 14 of 30 cases by STR-PCR and in 8 of 30 cases by FISH (p > 0.05). On day 28, the CC was detected in 26 of 31 cases by STR-PCR and in 15 of 31 cases by FISH (p < 0.01). At 3 months, the CC was observed in 22 of 24 cases by STR-PCR and 17 of 24 cases by FISH (p > 0.05). 14 cases were found to have a graft rejection or relapse among 28 cases which were continuously monitored more than 3 months post the transplants. Donor cell decrease in 9 of 14 cases was proved by FISH alone. It is concluded that FISH is more sensitive than STR-PCR in early monitoring chimerism status of post-transplant and in predicting graft rejection or disease relapse.

[The role of FDG-PET in staging of lymphoma and evaluation of therapeutic efficiency].

OBJECTIVE: To evaluate the application of 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) to the staging and detecting residual masses of lymphoma. METHODS: The clinical data of 179 patients with lymphoma were analyzed retrospectively. The results of FDG-PET, computed tomography (CT) and bone marrow biopsy (BMB) were compared for detection of lymph node/extranodal lymphoid tissue and bone marrow infiltration. Therapeutic efficiency was assessed by International Workshop Criteria (IWC) and Revised Integrated International Workshop Criteria (IWC + PET). RESULTS: In the detection of 286 disease focuses in 98 patients before chemotherapy, the sensitivities of FDG-PET and CT were 73% and 70% (P < 0.01) in detecting nodal focus,and 87% and 45% (P < 0.01) in detecting extranodal lymphoma respectively. In detection of 104 lesions in 81 patients after chemotherapy,the sensitivities of FDG-PET and CT were 81% and 55% respectively (P < 0.01), and the specificities were 68% and 33%, respectively (P < 0.01) in detecting residual masses. According to IWC criteria, 33 patients achieved complete response/unconfirmed complete response (CR/CRu) , and 8 (24%) relapsed. Patients with PET-positive residual masses had a relapse rate of 40%, whereas only 21% of those with no such masses relapsed. Based on IWC + PET criteria, 25 patients achieved CR, with a relapse rate of 20%. Both FDG-PET and BMB produced positive results in 133/179 (74%) patients. Twenty-two patients with positive FDG-PET results were not detected by BMB. The sensitivities and specificities of FDG-PET for BM infiltration were 52% and 83%, respectively. CONCLUSIONS: FDG-PET is a high sensitive and specific technique in staging and detecting residual masses of lymphoma.

[Allogeneic hematopoietic stem cell transplantation following reduced intensity conditioning regimen for treatment of refractory leukemia].

OBJECTIVE: To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) following reduced intensity conditioning (RIC) regimen for treatment of refractory leukemia. METHODS: Twenty patients with refractory leukemia received allo-HSCT following RIC regimen consisting of fludarabine plus small or moderate dose total body irradiation (TBI). Graft versus host disease (GVHD) prophylaxis was CsA plus mycophenolate mofetil (MMF) or short-term MTX, or these three drugs combination; CD25 monoclone antibody(McAb) and ATG were also used in some of the patients. RESULTS: Seventeen patients engrafted successfully, the median time for ANC > 0.5 x 10(9)/L was 13 (11 - 17) days, and for BPC > 50 x 10(9)/L 19 (12 -42) days. Detected by short tandem repeat (STR)-PCR, complete donor chimerism was confirmed in 16 patients with a median of 14 (7 -35) days. The incidence of acute and chronic GVHD was 47.1% (8/17) and 38.5% (5/13) respectively. The transplant related mortality (TRM) was 25.0% (5/20), mainly from graft failure, intracranial hemorrhage and severe infection. Up to now, 7 patients relapsed and 9 were alive with leukemia free. The overall survival (OS) at 2 year was (35. 3 +/- 14.2)% for all patients and (52.5 +/- 18.6)% for acute non-lymphocytic leukemia (ANLL) patients. CONCLUSION: Allo-HSCT following fludarabine and TBI based RIC regimen can be used for treatment of refractory leukemia with well tolerance and low TRM and there is a better prognosis for ANLL patients than that for acute lymphocytic leukemia patients.

[Clinical significance of dynamic monitoring of cell chimerism following allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To evaluate the relationship of chimerism status of cell subsets with engraftment, occurrence of acute graft versus host disease (aGVHD), graft rejection and disease relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: Chimerism status in peripheral blood (PB) and bone marrow (BM) of 65 patients received allo-HSCT were monitored at regular intervals post-transplant. Fluorescence-activated cell sorter (FACS) was used to sort CD3(+)T lymphocytes in 65 cases, CD3(-)CD56(+)CD16(+)NK cells in 52 cases, CD15(+) granulocytes in 32 cases and CD19(+)B lymphocytes in 20 cases post transplants. The chimerism status of different lineage cells was analyzed by polymerase chain reaction amplification of short tandem repeats (PCR-STR). RESULTS: On day +7, NK-cells donor chimerism (DC 55.5%) was higher than other cell subsets. T lymphocyte was the latest one to reach complete donor chimerism (CDC) with a median on day +21. Patients whose T lymphocytes donor chimerism was more than 70% on day +7 and more than 95% on day +14 had a high risk for acute aGVHD. In all cases except those with ALL, the decreased DC of T lymphocytes were observed before molecular or hematological relapse occurred. CONCLUSION: Serial and quantitative T cell chimerism analysis provides a reliable and rapid screening method for the early detection of engraftment, graft rejection, disease relapse and occurrence of aGVHD, therefore, is a prognostic tool to identify patients at high risk of aGVHD and disease relapse following allo-HSCT.

[Expression and analysis of biological activity of wild-type and mutant interleukin-13 in E.coli].

AIM: To express recombinant wild-type human interleukin-13 (rhIL-13) and mutant interleukin-13 (rhIL-13') in E.coli BL21 (DE3) and get active proteins through purification and renaturation. METHODS: IL-13 and IL-13' gene fragments were amplified by PCR and site-directed mutagenesis PCR, respectively and then were inserted into expression vector pET30a(+). Recombinant plasmids were transformed to E.coli BL21 (DE3) and were expressed under IPTG induction. Expressed products were purified through Ni-NTA chromatographic column. The purified proteins were renatured by GSH-GSSG (reduced glutathione, oxidized glutathione) system and dialysis and their bioactivity was detected by MTT colorimetry. RESULTS: The expressed recombinant proteins existed in the form of inclusion body with relative molecular mass about 17 000. The recombinant proteins with higher purity were obtained after purification. The renatured inclusion bodies were biologically active. CONCLUSION: rhIL-13/rhIL-13' with biological activity have been obtained successfully, which lays the foundation for further study on their function.

[Evaluation of the subsets of lymphocytes and their activated status in patients with myelodysplastic syndrome].

This study was purposed to investigate the clinical significance of the amount and activated status of T cell subsets, B cells, NK cells in peripheral blood from patients with myelodysplastic syndrome (MDS). The proportion of T cells, B cells, NK cells in peripheral blood from 30 patients with MDS and their surface activation markers of CD28, CD45RA, CD45RO, CD69, HLA-DR were analyzed by flow cytometry. Twenty-two patients were in the low risk group (RA + RAS) while eight patients were in the high risk group (RAEB + RAEBT). The result showed that the amounts of T cells (CD3+ cells) in peripheral blood from patients with MDS were lower than those in control group. The amounts of naive CD4+ cells (CD4+ CD45RA+ cells) in MDS patients were lower than those in control. The expression rates of early activation marker (CD69) and late activation marker (HLA-DR) on CD3+ cells in MDS patients were significantly higher than those in control. The abnormalities of the immunologically competent cells were mainly observed in the low risk group (RA + RAS), and were characterized by the high expression rates of CD69+ and HLA-DR+ on CD3+ cells, the decrease of B cell amounts. The amount abnormalities of T cell subsets were mainly observed in high risk group (RAEB + RAEBT), and were characterized by the decrease of CD3+ cells and CD3+ CD4+ CD8- cells (Th cells) amounts without high expression of the CD69 and HLA-DR, the decrease of NK cells amounts. It is concluded that there are the abnormalities of T cell subsets and function in the patients with MDS and may change with disease progression, so the measurement of amount and activated status of T cell subsets in peripheral blood from MDS patients can have predictive role for diagnosis of disease progression and guide of therapy.

[Transfection of chk1/2 antisense oligonucleotide to HL-60 cell line increases the apoptotic sensitivity to irradiation].

OBJECTIVE: To block signal transduction of cell cycle checkpoints by antisense blocking of chk1/2 gene to increase the radiation sensitivity of HL-60 cell line. METHOD: To transfect the HL-60 cell with chk1/2 antisense and sense chain alone and in combination, expose the cells to irradiation at 24 h after the transfection, the chk1 protein change was assayed by Western blot and the cell cycles and annexin V apoptosis rates by FCM. RESULTS: The irradiated apoptosis sensitivity was increased by antisense blocking of chk1 gene in HL-60 cell line, the apoptotic rate was 26.31% being significantly higher than that of the sense blocking (10.34%) (P < 0.05), Furthermore, the G(2)/M phase blocking phenomenon decreased and a synergic effect was observed in antisense blocking both the chk1 and chk2 genes. CONCLUSION: Antisense blocking of chk1/chk2 could increase the apoptotic sensitivity to irradiation.