[Effect of electroacupuncture on neurovascular unit and Wnt/ß-catenin signaling in rats with cerebral ischemia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Baihui" (GV20), "Shuigou" (GV26), etc. on the expressions of vascular endothelial growth factor (VEGF), collagen fibrillary acidic protein (GFAP), neuronal nucleus antigen(NeuN), ß-catenin and Axin2 protein and mRNA in rats with cerebral ischemia (CI), so as to explore its mechanism underlying improvement of ischemic stroke. METHODS: A total of 108 male SD rats were randomly divided into control, model and EA groups, which were further divided into 7 d, 14 d and 21 d subgroups, with 12 rats in each group. The CI model was established by occlusion of the middle cerebral artery. EA (2 Hz/100 Hz, 2-4 V) was applied to GV20, GV26, bilateral "Sanyinjiao" (SP6) and bilateral "Neiguan" (PC6) for 30 min, once daily (except Sundays) for 21 days at most. The neurological deficit score was evaluated according to Longa's methods. The cerebral infarction state was assessed by using a magnetic resonance T2 imaging system. The expression levels of neurovascular markers as VEGF,GFAP and NeuN, and ß-catenin and Axin2 protein and mRNA in the ischemic brain tissue were detected by using immunohistochemistry and quantitative real-time PCR, respectively. RESULTS: After modeling, the neurological deficit score and cerebral infarction size were significantly increased (P<0.01), and the expression of NeuN and Axin2 proteins and mRNAs were significantly and gradually decreased with time (day 7, 14 and 21) (P<0.01), whereas the expression levels of VEGF, GFAP, ß-catenin proteins and mRNAs were significantly increased on day 7, 14 and 21 in the model group relevant to the control group (P<0.01). Compared with the model group, the neurological deficit score, cerebral infarction size and the expressions of Axin2 protein and mRNA were significantly decreased on day 7, 14 and 21 (P<0.01), whereas the expression levels of VEGF, GFAP and NeuN and ß-catenin proteins and mRNAs were considerably up-regulated in the EA group on day 7, 14 and 21 (P<0.01). CONCLUSION: EA can protect the neurovascular units from injury, reduce the volume of cerebral infarction and improve the symptoms of neurological deficit in cerebral ischemic rats, which may be related to its effects in up-regulating ß-catenin expression and in down-regulating Axin2 expression to further activate classical Wnt/ ß-catenin signal pathway.

[Comparison of Decitabine Rigimen and Traditional Chemotherapy Regimen in Treatment of Patients with Intermediate or High-Risk Myelodysplastic Syndrome].

OBJECTIVE: To compare the clinical efficacy and relevant adverse reactions of homebred decitabine regimen and traditional chemotherapy regimen in treatment of patients with intermediate or high-risk myelodysplastic syndrome (MDS). METHODS: Forty-eight patients suffered from newly diagnosed intermediate or high-risk MDS from December 2011 to December 2016 were analyzed retrospectively. Among them 29 patients were treated by traditional chemotherapy regimen, and 19 patients were treated by decitabine regimen [15 mg/(m2·d), ivgtt, d1-5]. The clinical efficacy and relevant adverse reactions in two groups were compared. RESULTS: The overall response rate (ORR) of decitabine group was 78.9% (15/19), after 2 cycles of treatment, among them 5 achieved complete remission(CR), 5 achieved partial remission(PR), and 5 achieved hematologic improvement (HI); the ORR of traditional chemotherapy group was 65.9% (16/29), including 6 CR, 5 PR, 8 HI, the ORR and remission rate (PR+CR) in decitabine treatment group were not statistically significantly different from the that in traditional chemotherapy group (x2=0.458,P>0.05; x2=0.499, P>0.05). After 4 cycles of treatment, the ORR in decitabine group was 84.2% (16/19), including 5 CR, 9 PR and 2 HI. The ORR in traditional chemotherapy group was 68.9% (20/29), including 6 CR, 5 PR and 9 HI. The ORR of decitabine group was not statistically significantly different from the that in traditional chemotherapy (x2=0.726,P>0.05), but the remission rate was statistically significantly different(x2=4.534,P<0.05). The overall survival and progression-free survival in the decitabine group were different statistically significantly different from the traditional chemotherapy (P<0.05; P<0.01). The incidences of III-IV grades adverse reactions of hemoglobin, platelet and neutrophile in the patients treated with decitabine and traditional chemotherapy group were 52.6% and 79.3% (P>0.05), 57.9% and 86.2%(P>0.05), 84.2% and 96.6%(P>0.05), respectively. The infection rates were 26.3% and 79.3%(P<0.05), respectively. CONCLUSION: The homebred decitabine can effectively treat intermediate-or high-risk MDS, also can be well tolerated. So, it is worth to be clinically popularized.

[The clinical value of F-18 FDG PET/CT in patients with secondary hemophagocytic syndrome].

The aim of this study was to investigate the role of F-18 fluoro-2-deoxyglucose positron emission tomography/computed tomography (F-18 FDG PET/CT) in diagnosis and prognostic evaluation of secondary hemophagocytic syndrome (HPS). A total of 11 secondary HPS patients examined with 18F-FDG-PET/CT were retrospectively analyzed. The diagnostic value of F-18 FDG PET/CT for malignancy detection was assessed. The values of maximum standardized uptake value (SUV(max)) in spleen (SUVS(p)) and in bone marrow (SUVBM) were measured to analyze their relationship with various laboratorial parameters and clinical outcome of secondary HPS patients. The results showed that 4 out of the 11 patients had malignancies, the sensitivity, specificity and diagnostic accuracy of F-18 FDG PET/CT for malignancy detection were 100%, 66.7% and 75% respectively, the SUV(max) of spleen and bone marrow showed no significant correlation with laboratorial parameters, a maximum SUVS(p) of 3.10 and a maximum SUVBM of 3.47 were the optimal cutoffs for predicting patients' outcome, the increased uptake of F-18 FDG in the BM and spleen were significantly associated with shorter survival time according to univariate analysis. It is concluded that 18F-FDG PET/CT may especially play an important role in diagnosis and predicting outcome of secondary HPS for the small sample size.

[Protective effect of electroacupuncture intervention on neurovascular unit in rats with focal cerebral ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of electroacupuncture (EA) intervention on behavior changes, expression of cerebral vascular endothelial growth factor (VEGF), nerve growth associated protein-43 (GAP-43), synaptophysin (SYN), myelin basic protein (MBP), neurite outgrowth inhibitor-A (Nogo-A) in cerebral focal ischemia-reperfusion injury (CI/RI) rats, so as to study its mechanism underlying improvement of ischemic cerebral vascular disease. METHODS: Sixty male SD rats were randomly divided into sham-operation group, model group and electroacupuncture (EA) group. CI/RI model was established by occlusion of the middle cerebral artery (MCAO) and reperfusion. EA was applied to bilateral "Neiguan" (PC 6), "Sanyinjiao" (SP 6), "Shuigou" (GV 26) and "Baihui" (GV 20) for 30 min, once a day for 14 days. The neurologic deficits were evaluated by Longa 5-grade standard (the higher the score, the severer the neurologic deficit). The immunoactivity of cerebral VEGF, GAP-43, SYN, MBP (important in the process of myelination of nerves in the nervous system) and Nogo-A (inhibiting axonal regeneration) in the focal ischemic cerebral tissue was detected by immunohistochemistry. RESULTS: The Longa's score of the model group was significantly increased after MCAO in comparison with the sham-operation group (P < 0.01). In comparison with the model group, Longa's score of the EA group was evidently lower on day 14 after CI/RI (P < 0.05), suggesting an improvement of the neurological deficits after EA intervention. In comparison with the sham-operation group, the immunoactivity of cerebral VEGF, GAP-43 and Nogo-A was significantly upregulated on day 7 and 14 in the model group (P < 0.01), while that of cerebral SYN was remarkably down-regulated in the model group on day 7 and 14 after CI/RI (P < 0.05). Compared with the model group, cerebral VEGF, GAP-43, SYN and MBP expression levels were considerably upregulated on day 7 and 14 following CI/RI in the EA group (P < 0.01, P < 0.05), while that of cerebral Nogo-A was significantly decreased at the two time-points in the EA group (P < 0.01). CONCLUSION: EA intervention can effectively improve neurological function in cerebral infarction rats, which is closely related to its effects in upregulating cerebral VEGF, GAP-43, SYN and MBP expression, and down-regulating Nogo-A protein, suggesting a protective effect on neurovascular unit.

[Immunophenotyping and molecular genetic analysis of diffuse large B-cell lymphoma].

OBJECTIVE: To perform immunophenotyping and molecular genetic analysis for diffuse large B-cell lymphoma (DLBCL), and to explore their correlation and implication for prognosis. METHODS: Immunohistochemical streptavidin peroxidase (SP) method was used to determine the expression of CD10, BCL6 and MUM1 in 59 cases of DLBCL. A Hans algorithm was used to classify DLBCL into germinal center B-cell (GCB) and non-GCB subtypes. Interphase fluorescence in situ hybridization (FISH) assay was performed on paraffin-embedded lymphoma tissue sections to detect translocations and amplifications of BCL6, BCL2 and MYC genes with dual-color break-apart BCL6 probe, dual-color dual-fusion IgH/ BCL2 probe and dual-color break-apart MYC probe, respectively. RESULTS: In the 59 cases of DLBCL, 28.8% (17/59) belonged to GCB subtype, and 71.2% (42/59) belonged to non-GCB subtype. The incidences of BCL6, BCL2 and MYC gene translocations were 24.1% (14/58), 1.7% (1/59) and 5.3% (3/57), respectively. The incidences of BCL6, BCL2 and MYC gene amplifications were 17.2% (10/58), 22.0% (13/59) and 21.1% (12/57), respectively. BCL6 amplification was not correlated with BCL6 translocation (P=0.424), but was correlated with amplifications of BCL2 and MYC (C=0.405 and 0.403, respectively, P <0.01). The incidence of BCL6 translocation in GCB type was higher than that in non-GCB type, and amplifications of BCL6, BCL2 or MYC were more frequently encountered in non-GCB type, though no statistical significance was detected (P=0.089 and 0.106, respectively). By univariate analysis, immunophenotyping and international prognostic index (IPI) exerted a significant effect on overall survival (OS) (P=0.047 and 0.001, respectively), but to which BCL6 translocation and amplification of the 3 genes were not related (P=0.150 and 0.444, respectively). By multivariate analysis, IPI score was the only independent prognostic factor for OS (RR =3.843, P=0.017). CONCLUSION: The GCB subtype of DLBCL is less common in the patient cohort. Common genetic aberrations have included BCL6 translocation and BCL6, BCL2 and MYC amplifications. Amplification of the 3 genes is strongly correlated with each other, and the incidence of BCL2 translocation is low. Immunophenotyping only has minor significance for the prognosis. Genetic aberrations cannot predict the clinical outcome of DLBCL.

[Clinical and molecular cytogenetic studies of a case of B-lineage acute lymphoblastic leukemia with t(14;14)(q11;q32)].

OBJECTIVE: To report on a rare case of B-lineage acute lymphoblastic leukemia (B-ALL) with t(14;14) (q11;q32) and clarify its clinical and molecular cytogenetic features. METHODS: Clinical data of a B-ALL patient with t(14;14) (q11;q32) were analyzed. After 24 hour of unstimulated culturing, chromosome specimens of bone marrow cells were prepared with regular method, and R-banding was used for karyotype analysis. Fluorescence in situ hybridization (FISH) analysis was performed on fixed bone marrow cells using IGH dual-color break-apart probe, CEBPE dual-color break-apart probe, whole chromosome paint (WCP) probe for chromosome 4, and Chromoprobe Multiprobe-ALL System probe. RESULTS: The 39-year-old female was diagnosed with B-ALL based on morphologic and immunophenotypic analyses. Conventional cytogenetic analysis showed a karyotype of 47, XX, +4, t(14;14) (q11;q32) [20], which was confirmed by FISH analysis. FISH using IGH-dual-color break-apart probe confirmed involvement of IGH gene in t(14;14) (q11;q32), and FISH using CEBPE dual-color break-apart probe indicated that CEBPE is the partner gene involved in t(14;14) (q11; q32). The patient achieved complete remission (CR) after a round of combined chemotherapy. At the time of follow-up, she had remained CR for more than 6 months. CONCLUSION: t(14;14) (q11;q32) simultaneously involving IGH and CEBPE genes in B-ALL is a rare but recurrent genetic abnormality that may identify a new subgroup of B-ALL. In B-ALL patients, t(14; 14) (q11; q32) involving IGH/CEBPE translocation may indicate a better prognosis.

[Report of ten cases of hematologic malignancies with idic(20q-) and literature review].

OBJECTIVE: To analyze the clinical and molecular cytogenetic features of hematologic malignancies with idic(20q-). METHODS: The clinical data of 10 patients with idic (20q-) were analyzed. Karyotyping analysis was carried out with R banding technique. A CEP20 probe was used to perform single-color fluorescence in situ hybridization (FISH). A subtelomeric probe for 20q and a locus-specific probe for 20q12 were used to perform dual-color FISH. The literatures of hematologic malignancies with idic(20q-) were reviewed. RESULTS: Of the 10 cases, 2 were diagnosed as acute erythroid leukemia, 1 primary myelofibrosis, 3 myelodysplastic syndromes (MDS) and 4 highly suspected (HS-MDS). Karyotype analysis showed that one of the normal chromosome 20 allele was substituted by one or two metacentric isochromosomes smaller than the normal one in all 10 cases. It was confirmed to be der(20)del(20)(q11q13)idic(20)(p11), i.e., idic(20q-) by FISH assay. Partial cells in 2 of the 10 cases had 20q- as the sole karyotypic anomaly. CONCLUSION: Idic(20q-) results from a pre-existing del(20q) and is strongly associated with MDS and acute erythroid leukemia. Idic(20q-) as a recurrent cytogenetic abnormality is helpful for diagnosing HS-MDS in patients with cytopenia but only slight or absent dysplasia.

[Unrelated cord blood transplantation in adult patients with hematologic malignancies].

OBJECTIVE: To analyse the engraftment, transplant-related complications and survival after unrelated cord blood transplantation (UCBT) in patients with hematologic malignancies. METHODS: Twenty eight consecutive adult patients with hematological malignancies were treated with UCBT and 20 of them were advanced-stage diseases. Double or multiple UCB grafts were used for 18 patients, while single UCB graft for 10 patients. Myeloablative conditioning regimens were given to 26 cases and nonmyeloablative regimens to 2 cases. All patients were given a combination of cyclosporine (CsA) and mycophenolate mofetil (MMF) for graft-versus-host disease (GVHD) prophylaxis. RESULTS: Median time to neutrophil engraftment (≥ 0.5 × 10(9)/L) in 26 patients was 18 (14 - 37) days and platelet engraftment (≥ 20 × 10(9)/L) in 22 patients was 30 (25 - 49) days. Chimerism was weekly assessed by PCR analysis of short tandem repeat (STR) sequences in whole blood or bone marrow and 22 cases were confirmed of fully donor chimeric from 7 to 21 days after transplantation. Eighteen cases developed acute GVHD, greater than grade II in 1, and 6 of 22 patients who survived more than 100 days developed limited chronic GVHD. Eighteen cases were alive in hematologic remission at a median follow-up of 9.5 (2.5 - 72.0) months. The probability of event-free survival at 3 years was 56.7%. Two cases relapsed and 8 of 10 cases died of transplant related complications. CONCLUSIONS: UCBT could be safely and effectively used for adult patients with hematologic malignancies. Use of double UCB units is a strategy extending the feasibility of UCBT.

HLA-matched sibling transplantation with G-CSF mobilized PBSCs and BM decreases GVHD in adult patients with severe aplastic anemia.

BACKGROUND: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for severe aplastic anemia (SAA). However, graft failure and graft-versus-host disease (GVHD) are major causes of the early morbidity in Allo-HSCT. METHODS: To reduce graft failure and GVHD, we treated fifteen patients with SAA using high- dose of HSCT with both G-CSF mobilized PB and BMSCs from HLA-identical siblings to treat patients with SAA. RESULTS: All patients had successful bone marrow engraftment. Only one patient had late rejection. Median time to ANC greater than 0.5 × 10(9)/L and platelet counts greater than 20 × 10(9)/L was 12 and 16.5 days, respectively. No acute GVHD was observed. The incidence of chronic GVHD was 6.67%. The total three-year probability of disease-free survival was 79.8%. CONCLUSION: HSCT with both G-CSF mobilized PB and BMSCs is a promising approach for heavily transfused and/or allo-immunized patients with SAA.