Clinical risk score for invasive fungal diseases in patients with hematological malignancies undergoing chemotherapy: China Assessment of Antifungal Therapy in Hematological Diseases (CAESAR) study / 医学前沿

Invasive fungal disease (IFD) is a major infectious complication in patients with hematological malignancies. In this study, we examined 4889 courses of chemotherapy in patients with hematological diseases to establish a training dataset (n = 3500) by simple random sampling to develop a weighted risk score for proven or probable IFD through multivariate regression, which included the following variables: male patients, induction chemotherapy for newly diagnosed or relapsed disease, neutropenia, neutropenia longer than 10 days, hypoalbuminemia, central-venous catheter, and history of IFD. The patients were classified into three groups, which had low (0-10, ~1.2%), intermediate (11-15, 6.4%), and high risk ( > 15, 17.5%) of IFD. In the validation set (n = 1389), the IFD incidences of the groups were ~1.4%, 5.0%, and 21.4%. In addition, we demonstrated that antifungal prophylaxis offered no benefits in low-risk patients, whereas benefits were documented in intermediate (2.1% vs. 6.6%, P = 0.007) and high-risk patients (8.4% vs. 23.3%, P = 0.007). To make the risk score applicable for clinical settings, a pre-chemo risk score that deleted all unpredictable factors before chemotherapy was established, and it confirmed that anti-fungal prophylaxis was beneficial in patients with intermediate and high risk of IFD. In conclusion, an objective, weighted risk score for IFD was developed, and it may be useful in guiding antifungal prophylaxis.

Effect of endothelial cell-targeted soluble Notch ligand hD1R protein on the proliferation of acute myeloid leukemia cells / 中华血液学杂志

Objective@#To evaluate the effects of endothelial cell-targeted soluble Notch ligand hD1R protein on the proliferation of acute myeloid leukemia (AML) cells.@*Methods@#The expression levels of Notch1, Notch2, Notch3, Notch4, Hes1 in bone marrow CD34+ cells from 24 cases of untreated AML (AML group) and 9 healthy controls (control group) were determined by real time quantitative polymerase chain reaction (PCR) . Recombinant hD1R protein was first induced and purified. Bone marrow CD34+ cells were co-cultured on human umbilical vein endothelial cells (HUVEC) supplemented with a cocktail containing 5 types of human cytokines (5GF) and soluble hD1R. The cultured cells were tested under different culture conditions including PBS group (PBS replaces HUVEC) , hD1R group, 5GF group, GSI group (hD1R plus GSI) . Proliferation and apoptosis of cultured cells were also analyzed. Real time quantitative PCR was used to test the expression levels of Hes1 and Bcl-2 in cultured cells.@*Results@#The expression levels of Notch1 and Hes1 in primary AML patients were significantly lower, and Notch4 expression was higher compared to the control group (P<0.05) . Cell counting showed a remarkable decrease of AML cells number in the culture with hD1R compared with that in the PBS group[ (0.74±0.13) ×105 vs (2.16±0.21) ×105, P<0.01]. FACS analysis showed that the percentage of AML cells was (18.48±2.51) % in apoptosis, which was higher than that of control cells (3.19±0.58) % after co-culture with hD1R. AML cells in the hD1R group underwent significantly increased apoptosis compared with those in the PBS one (P<0.05) . Moreover, apoptosis of AML cells in the GSI group was lower than that in the hD1R one (P<0.05) . Apoptosis in the PBS group also decreased compared with that in the 5GF one (P<0.05) . Finally, hD1R up-regulated Hes1 expression and inhibited Bcl-2 one in the AML cells.@*Conclusion@#hD1R effectively activated Notch signaling and down-regulated Bcl-2 mRNA in AML cells, which lead to cell apoptosis.

Efficacy of low-dose cytarabine and harringtonine regimen as induction in different risk stratifications of acute myeloid leukemia / 白血病·淋巴瘤

Objective To explore the clinical efficacy of low-dose cytarabine and harringtonine (LD-HA) regimen in the induction therapy of acute myeloid leukemia (AML) except M3. Methods 52 AML patients who received LD-HA were analyzed retrospectively. The patients were graded according to molecular biological and cytogenetic risk degree. The clinical efficacy, toxicity of LD-HA and long-term survival followed-up were compared with those of idarubicin and cytarabine (IA) regimen in 49 patients. Results After one cycle, the overall remission (OR) rates of LD-HA group and IA group were 71.2%(37/52) [CR rate 50.0%(26/52), PR rate 21.2%(11/52)] and 53.1%(26/49) [CR rate 44.9%(22/49), PR rate 8.2%(4/49)], respectively, with no statistical significance of OR between the two groups (P= 0.068). OR rates were not statistically significant in either low-risk group or intermediate-risk group between LD-HA group and IA group (P> 0.05), but OR of high-risk group in LD-HA was much higher than that in IA group [100 % (11/11) vs 66.7 % (12/18), P<0.05]. Cardiac toxicity and bone marrow suppression in LD-HA group were much milder than those in IA group. The patients unfit for standard chemotherapy could tolerate to LD-HA regimen. Conclusions LD-HA regimen as induction for high risk AML patients can improve the OR rate, and reduce the side effects, which is beneficial for high-risk AML patients.

Clinical features of rituximab plus chemotherapy as first-line treatment in patients with diffuse large B-cell lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>A prospective, multicenter and non-interventional prospective study was conducted to evaluate the clinical features of rituximab combined with chemotherapy (R-Chemo) as first-line treatment on newly diagnosed Chinese patients with diffuse large B-cell lymphoma (DLBCL).</p><p><b>METHODS</b>This was a single arm, prospective, observational multicenter and phase IV clinical trial for 279 patients, who were newly diagnosed as CD20-positive DLBCL from 24 medical centers in China 2011 and 2012, no special exclusion criteria were used. All patients received rituximab based R-Chemo regimes, such as R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisolone) and other regimes as the first-line treatment. The treatment strategies were determined by physicians and patients without detailed description for treatment course, dose, interval time and examination. Clinical response and safety of all patients were investigated in 120 days after completion of last dose of rituximab.</p><p><b>RESULTS</b>Of 279 patients, 258 with stage I-IV who received at least 1 cycle of rituximab treatment and completed at least one time of tumor assessment were enrolled into intention-to-treat analysis, including 148 male and 110 female. The median age of all patients was 57.2(12.8-88.4) years. ECOG performance statuses of 0 or 1 were observed in 91.1% of patients, international prognostic index levels in the low-risk and low-middle-risk groups in 76.4% of patients, the tumor diameters smaller than 7.5 cm in 69.0% of patients. All patients received 6 median cycles of R-Chemo treatment every 24.4 days. R-CHOP treatment was shown to improve the clinical response with overall response rates of 94.2%. Common adverse events included anemia, marrow failure, leukopenia, thrombocytopenia, digestive diseases, infection and liver toxicity. All adverse events are manageable.</p><p><b>CONCLUSION</b>Non-interventional clinical trial of R-Chemo remains the standard first-line treatment for newly diagnosed patients with DLBCL in real clinical practice, which is consistent with international treatment recommendations for DLBCL patients. R-Chemo can provide the clinical evidence and benefit as the first-line standard treatment for Chinese patients with DLBCL.</p>

Effect of endothelial cell-targeted soluble Notch ligand hD1R protein on expansion and engraftment of cord blood hematopoietic stem/progenitor cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of endothelial cell- targeted soluble Notch ligand hD1R protein on expansion and engraftment of cord blood hematopoietic stem/progenitor cell (CB HSPCs).</p><p><b>METHODS</b>Recombinant hD1R protein was first induced and purified. Human cord blood CD34⁺ cells were co-cultured on human umbilical vein endothelial cells (HUVECs) supplemented with a cocktail containing 5 types of human cytokines including TPO, SCF, FL, IL-6, IL-3 (5GF) and soluble hD1R. The expansion of CD34⁺ cells was tested under different culture conditions including PBS group (PBS replaces HUVEC), hD1R group, sup group (HUVEC supernatant replaces HUVEC), fix group (fixed HUVEC replaces HUVEC), Day 0 group (Control). Cell cycle and apoptosis of cultured cells were also analyzed. Their progeny expanded in PBS or hD1R group were transplanted into sublethally irradiated NOD/SCID mice. The percentages of human CD45⁺ (hCD45⁺) cells in the marrow of recipient mice were determined by FACS 12 weeks later.</p><p><b>RESULTS</b>hD1R induced more expansion in the total number of CD34⁺ cells cocultured with HUVECs plus 5GF, which was 87.50-fold increase compared to the Day 0 group, and 7.98-fold increase than that of PBS group. FACS analysis also showed that the percentage of CD34⁺ cells was 77.0% in G0/G1 phase in the hD1R group, which indicated that hD1R enhanced HSPCs expansion and inhibited apoptosis. Moreover, hD1R significantly promoted human HSPC engraftment after BM transplantation in irradiated mice.</p><p><b>CONCLUSION</b>The Notch-mediated ex vivo expansion system has been established and hD1R promoted expansion and engraftment of human CB HSPCs, which provided the evidence for further clinical application.</p>

Treatment to high-risk acute non-lymphocytic leukemia with sequential induction / 白血病·淋巴瘤

Objective To investigate the outcome of high-risk acute non-lymphocytic leukemia treated with sequential low-dose cytarabine and harringtonine(LD-HA) and standard induction. Methods 50 high-risk ANLL. patients (LD-HA group) who were regarded as unfit for intensive chemotherapy were chosen to receive LD-HA. Reinductive treatments with standard regimens would be given for those who did not achieve complete remission. 23 patients DA/HA group given two cycles of standard inductive regimens were taken as the control. Results In LD-HA group 80.0 %. (40/50) reached CR, 2 patients died shortly after inductive therapy. The median leukemia-free survival(LFS) was 19.6 months, and the median overall survival (OS) was 12.2 months. Overall survival was 57.0 % at 1 year, 24.1% at 3 years, and 18.8 % at 5 years. While the CR rate was 73.9 % for DA/HA group, and none died during the inductive therapy. LFS and OS was 19.8 months and 12.1 months, respectively. OS rate was 56.58 % at 1 year, 27.1% at 3 years, and 27.1% at 5 years.There were no difference on OS rates between 2 groups (x2 were 0.009, 0.237 and 1.807, respectively,P ＞0.05). Conclusion In patients who were unfit for intensive chemotherapy, sequential therapy with LD-HA and standard induction improved the rate of complete remission and the duration of survival.

Construction and expression of a vector containing protein transduction domain and bcr/abl fusion gene / 中华血液学杂志

<p><b>OBJECTIVE</b>To construct a vector containing protein transduction domain (PTD) and bcr/abl fusion gene of chronic myelogenous leukemia and express PTD-bcr/abl fusion protein in E. Coli.</p><p><b>METHODS</b>DNA fragment encoding PTD was synthesized and fused to PCR-amplified bcr/abl gene fragment, then inserted into plasmid pET-16b to get the expression vector pEPb containing PTD-bcr/abl fusion gene, which was transfected and expressed in E. Coli LB21. PTD-bcr/abl fusion protein was purified by affinity chromatography.</p><p><b>RESULTS</b>523 bp bcr/abl fusion gene was effectively amplified. The PTD-bcr/abl gene sequencing showed the same sequence as scheduled. The fusion peptide was successfully expressed in E. Coli and purified.</p><p><b>CONCLUSION</b>The results may provide a new PTD-bcr/abl fusion peptide for the immunotherapy of CML.</p>

Expression of PTD-bcr/abl fusion oncoprotein fragment carrying protein transduction domain and its transmembrane transportation / 细胞与分子免疫学杂志

Aim To explore the method of transfering oncoprotein bcr/abl, mediated by protein transduction domain (PTD), in order to provide the experimental basis for immune therapy. Methods DNA fragment encoding PTD was synthesized and fused with PCR-amplified bcr/abl gene fragment. The fusion protein was expressed in E.coli as His-tagged protein. The purified fusion protein was loaded to cultured HL-60 cells. Whether the fusion protein entered into HL-60 cells was determined by Western blot. Results It is shown that PTD could mediate transportation of bcr/abl protein to enter into cells. Conclusion These results may provide a new approach for loading exogenous proteins to antigen presenting cells.