[Clinical Efficiency of Rituximab Combined with Autologous Hematopoietic Blood Stem Cell Transplantation for Treatment of CD20+ B non.Hodgkin Lymphoma].

AbstractObjective: To investigate the effeciency of autologous hematopoietic stem cell transplantation (auto.HSCT) combined with rituximab(R) to treat CD20+ B non.Hodgkin lymphoma(B.NHL). METHODS: From January 2005 to December 2013, 83 patients with refractory/recurrent CD20+ B.NHL who were treated with auto.HSCT in our department were enrolled. The patients were divided into 2 groups: 57 patients in Rituximab group, and 26 patients in control group(without Rituximab). All the patients received chemotherapy and auto.HSCT. For the patients in treatment group, Tituximab was used before transplantation of the stem cells, and for some patients Rituximab was used after transplantation. For the patients in control group, the induction, enhancement and transplantation were the same as those in treatment group. The clinical efficiency of the patients in treatment group according to the time and frequency of R was analyzed in subgroups and compared with the control group. The deadline of follow.up was April 30 2014. RESULTS: All the patient achieved complete response. The median follow.up time was 39 months. Both the two groups collected peripheral blood stem cells successfully, and had no difference in hematopoietic reconstitution time. Three patients in treatment group and six patients in control group relapsed and the three year overall survival and EFS in treatment group was significantly higher than that in control group, that is（93.0% vs 73.1%, P=0.037） and （89.5% vs 65.4%, P=0.034）, respectively. Subgroup analysis showed that: compared with the treatment group in which using R in the whole courses(before and after transplantation, and collection of stem cells) was superior to the control group in both OS and EFS, with the OS 97% vs 87.5% (P>0.05) and EFS 97% vs 76.2% (P=0.05) respectively. While stratified by the different courses of rituximab, the OS was 88.9% (1-2 courses, 9 cases), 93.1% (3-4 courses, 29 cases), 94.7%(more than 5 courses,19 cases), and EFS was 77.8%, 89.7% and 94.7%, respectively. CONCLUSION: For the patients with refractory/recurrent CD20+ B.NHL, the combination of R and inducing chemotheraphy, purify in body before transplantation, as well as continue with R after auto.HSCT could obviously improve the OS and EFS of patients. For the patients who with R before and after transplantation, their EFS is better than the patients with R before transplantation only.

Endophthalmitis associated with fungal keratitis and penetrating injuries in North China.

PURPOSE: To report the laboratory findings, management strategies, and visual outcomes of culture-proven exogenous fungal endophthalmitis in North China. METHODS: The microbiological and treatment records of patients with culture-positive exogenous fungal endophthalmitis who visited the Affiliated Hospital of Qingdao University from January 2012 to December 2016 were reviewed. RESULTS: A total of 39 eyes (39 patients) were identified over a 5-year period. Exogenous fungal endophthalmitis was associated with penetrating trauma in 22 eyes (56.4%), fungal keratitis in 15 eyes (38.5%), and intraocular surgery in 2 eyes (5.1%). Hyphae were found in 29 of 37 smear samples (78.4%) by direct microscopic examination. Fungal pathogens cultured from 39 samples were identified as 10 genera and 15 species. Filamentous fungi (molds) accounted for 94.9% (37 samples), including Fusarium (19, 48.7%) and Aspergillus (11, 28.2%). Most keratitis cases were caused by Fusarium (11 of 15; 73.3 %). Aspergillus was isolated from nine penetrating ocular trauma cases (9 of 22; 40.9%). Three eyes receiving evisceration had fungal and bacteria coinfection (3 of 39, 7.7%) with Aspergillus and Bacillus. At least, one surgical intervention was performed in all 39 eyes and 28 (71.8%) eyes underwent two or more procedures, including surgeries and intraocular injections. Twenty-nine patients received intraocular antifungal therapy with amphotericin B and/or voriconazole. Visual acuity at discharge from the hospital was significantly better than the initial visual acuity (p < 0.001). Final vision of 20/400 or better was achieved in 22 (56.4%) eyes. CONCLUSIONS: This study highlighted the differences between clinical categories of exogenous fungal endophthalmitis. Trauma was the major etiological factor. Molds were the most common pathogens, with Fusarium ranking first, followed by Aspergillus. Fungal and bacterial coinfection mostly occurred after metal penetrating trauma, and Bacillus was the primary bacterial pathogen. Coinfection may be one reason of evisceration. Immediate intravitreal antifungal therapy combined with vitrectomy was effective for exogenous fungal endophthalmitis. Amphotericin B and voriconazole were commonly used antifungal agents.

lncRNA PCAT-1 interacting with FZD6 contributes to the malignancy of acute myeloid leukemia cells through activating Wnt/ß-catenin signaling pathway.

Accumulating evidence has suggested the involvement of long noncoding RNAs (lncRNAs) on the acute myeloid leukemia (AML). Therefore, this study aimed to investigate the unknown function of lncRNA Prostate cancer-associated transcript-1 (PCAT-1) in AML cells. Our data found that PCAT-1 was highly expressed in AML-M1/2 and AML-M3 patients than normal controls and its expression was significantly up-regulated in AML cell lines Kasumi-6 and HL-60. The functional experiments demonstrated that knockdown of PCAT-1 remarkably inhibited proliferation, arrested cell cycle progression and triggered apoptosis of AML cells. Mechanistically, we revealed that PCAT-1 could directly interact with FZD6 protein to regulate its stability. Overexpression of FZD6 partly abolished the effects of PCAT-1 silencing on AML cells. Our integrated experiments then suggested that PCAT-1 could activate the Wnt/ß-catenin signaling pathway in an FZD6-dependent manner. Taken together, the present study indicated that PCAT-1 interacting with FZD6 to activate Wnt/ß-catenin signaling, which may play an important role in the pathogenesis of AML.

c-Myc suppresses miR-451⊣YWTAZ/AKT axis via recruiting HDAC3 in acute myeloid leukemia.

Aberrant activation of c-Myc plays an important oncogenic role via regulating a series of coding and non-coding genes in acute myeloid leukemia (AML). Histone deacetylases (HDACs) can remove acetyl group from histone and regulate gene expression via changing chromatin structure. Here, we found miR-451 is abnormally down-regulated in AML patient samples; c-Myc recruits HDAC3 to form a transcriptional suppressor complex, co-localizes on the miR-451 promoter, epigenetically inhibits its transcription and finally induces its downregulation in AML. Furthermore, our in vitro and in vivo results suggest that miR-451 functions as a tumor suppressor via promoting apoptosis and suppressing malignant cell proliferation. The mechanistic study demonstrated that miR-451 directly targets YWHAZ mRNA and suppresses YWHAZ/AKT signaling in AML. Knockdown of c-Myc results in restoration of miR-451 and inhibition of YWHAZ/AKT signaling. In AML patients, low level of miR-451 is negatively correlated with high levels of c-Myc and YWHAZ, while c-Myc level is positively related to YWHAZ expression. These results suggested that c-Myc⊣miR-451⊣YWHAZ/AKT cascade might play a crucial role during leukemogenesis, and reintroduction of miR-451 could be as a potential strategy for AML therapy.

The PU.1-Modulated MicroRNA-22 Is a Regulator of Monocyte/Macrophage Differentiation and Acute Myeloid Leukemia.

MicroRNA-22 (miR-22) is emerging as a critical regulator in organ development and various cancers. However, its role in normal hematopoiesis and leukaemogenesis remains unclear. Here, we detected its increased expression during monocyte/macrophage differentiation of HL-60, THP1 cells and CD34+ hematopoietic stem/progenitor cells, and confirmed that PU.1, a key transcriptional factor for monocyte/macrophage differentiation, is responsible for transcriptional activation of miR-22 during the differentiation. By gain- and loss-of-function experiments, we demonstrated that miR-22 promoted monocyte/macrophage differentiation, and MECOM (EVI1) mRNA is a direct target of miR-22 and MECOM (EVI1) functions as a negative regulator in the differentiation. The miR-22-mediated MECOM degradation increased c-Jun but decreased GATA2 expression, which results in increased interaction between c-Jun and PU.1 via increasing c-Jun levels and relief of MECOM- and GATA2-mediated interference in the interaction, and thus promoting monocyte/macrophage differentiation. We also observed significantly down-regulation of PU.1 and miR-22 as well as significantly up-regulation of MECOM in acute myeloid leukemia (AML) patients. Reintroduction of miR-22 relieved the differentiation blockage and inhibited the growth of bone marrow blasts of AML patients. Our results revealed new function and mechanism of miR-22 in normal hematopoiesis and AML development and demonstrated its potential value in AML diagnosis and therapy.

ZFP36L1 promotes monocyte/macrophage differentiation by repressing CDK6.

RNA binding proteins (RBPs)-mediated post-transcriptional control has been implicated in influencing various aspects of RNA metabolism and playing important roles in mammalian development and pathological diseases. However, the functions of specific RBPs and the molecular mechanisms through which they act in monocyte/macrophage differentiation remain to be determined. In this study, through bioinformatics analysis and experimental validation, we identify that ZFP36L1, a member of ZFP36 zinc finger protein family, exhibits significant decrease in acute myeloid leukemia (AML) patients compared with normal controls and remarkable time-course increase during monocyte/macrophage differentiation of PMA-induced THP-1 and HL-60 cells as well as induction culture of CD34(+) hematopoietic stem/progenitor cells (HSPCs). Lentivirus-mediated gain and loss of function assays demonstrate that ZFP36L1 acts as a positive regulator to participate in monocyte/macrophage differentiation. Mechanistic investigation further reveals that ZFP36L1 binds to the CDK6 mRNA 3'untranslated region bearing adenine-uridine rich elements and negatively regulates the expression of CDK6 which is subsequently demonstrated to impede the in vitro monocyte/macrophage differentiation of CD34(+) HSPCs. Collectively, our work unravels a ZFP36L1-mediated regulatory circuit through repressing CDK6 expression during monocyte/macrophage differentiation, which may also provide a therapeutic target for AML therapy.

Nutlin-3-induced redistribution of chromatin-bound IFI16 in human hepatocellular carcinoma cells in vitro is associated with p53 activation.

AIM: Interferon-γ inducible protein 16 (IFI16), a DNA sensor for DNA double-strand break (DSB), is expressed in most human hepatocellular carcinoma cell (HCC) lines. In this study we investigated the re-localization of chromatin-bound IFI16 by Nutlin-3, a DNA damage agent, in HCC cells in vitro, and the potential mechanisms. METHODS: Human HCC SMMC-7721 (wild-type TP53), Huh-7 (mutant TP53), Hep3B (null TP53) and normal fetal liver L02 cell lines were examined. DSB damage in HCC cells was detected via γH2AX expression and foci formation assay. The expression of IFI16 and IFNB mRNA was measured using RT-PCR, and subcellular localization and expression of the IFI16 protein were detected using chromatin fractionation, Western blot analysis, and fluorescence microscopy. RESULTS: Treatment of SMMC-7721 cells with Nutlin-3 (10 µmol/L) or etoposide (40 µmol/L) induced significant DSB damage. In SMMC-7721 cells, Nutlin-3 significantly increased the expression levels of IFI16 and IFNB mRNA, and partially redistributed chromatin-bound IFI16 protein to the cytoplasm. These effects were blocked by pretreatment with pifithrin-α, a p53 inhibitor. Furthermore, Nutlin-3 did not induce ectopic expression of IFI16 protein in Huh-7 and Hep3B cells. Moreover, the association of IFI16 with chromatin and Nutlin-3-induced changes in localization were not detected in L02 cells. CONCLUSION: Nutlin-3 regulates the subcellular localization of IFI16 in HCC cells in vitro in a p53-dependent manner.

Hemophagocyte lymphohistiocytosis secondary to bilateral epididymal diffuse large B-cell lymphoma.

We report a rare case of hemophagocytic lymphohistiocytosis (HLH) secondary to bilateral epididymal lymphoma. The patient is a man of 46 years old. He suffered fever 5 months before the admission. Physical examination shows splenohepatomegalia without lymphadenopathy. Laboratory studies revealed pancytopenia and coagulopathy, hyper ferritin level, large foamy macrophages containing other hematopoietic elements in bone marrow. After treatment with HLH-04 protocol, the patient recovered, but relapsed 1 month later and suffered pudendal pain. A color Doppler of the scrotum shows bilateral epididymis swollen. Pathological diagnosis of biopsy is diffuse large B cell lymphoma (DLBCL). After two cycles cyclophosphamide, doxorubicin, vincristine, and prednisone plus rituximab together with intrathecal injections, the tumors on the epididymides disappeared, but relapsed intracalvarium. The patient stopped treatment due to financial reasons. Hemophagocytic lymphohistiocytosis secondary lymphoma in a sanctuary site has not been reported before and should not be ignored. Transplantation is necessary for long-time survival.

Splenectomy improves anaemia but does not reduce iron burden in patients with haemoglobin H Constant Spring disease.

BACKGROUND: Splenectomy is reported to increase the haemoglobin level in patients with haemoglobin H Constant Spring (HbH CS) disease; however, its impact on iron burden and the underlying mechanism remains unclear. MATERIALS AND METHODS: From March through to May 2013, a total of 50 adults with HbH CS disease (25 cases splenectomised and 25 cases non-splenectomised) were enrolled. The patients' general conditions, history of blood transfusion and iron chelator treatment were investigated. Levels of haemoglobin, nucleated red blood cell counts, and serum ferritin were measured. The percentage of apoptotic erythroid precursor cells in bone marrow, an index representing ineffective erythropoiesis, was determined in some cases. RESULTS: There were no significant differences in age, blood transfusion volume, and use of iron chelator drugs between the splenectomised group and the non-splenectomised group. Significantly higher haemoglobin levels, serum ferritin levels and nucleated red blood cell counts as well as a higher percentage of apoptotic erythroid progenitor cells were detected in the splenectomised group. Regression analysis revealed that age and nucleated red blood cell counts were independent risk factors affecting the serum ferritin level. DISCUSSION: Despite improving the haemoglobin level, splenectomy is associated with greater iron burden in HbH CS disease. A high nucleated red blood cell count is predictive of the risk of severe iron overload.

MicroRNA-29a and microRNA-142-3p are regulators of myeloid differentiation and acute myeloid leukemia.

Although microRNAs (miRNAs) are increasingly linked to various physiologic processes, including hematopoiesis, their function in the myeloid development is poorly understood. We detected up-regulation of miR-29a and miR-142-3p during myeloid differentiation in leukemia cell lines and CD34(+) hematopoietic stem/progenitor cells. By gain-of-function and loss-of-function experiments, we demonstrated that both miRNAs promote the phorbol 12-myristate 13-acetate-induced monocytic and all-trans-retinoic acid-induced granulocytic differentiation of HL-60, THP-1, or NB4 cells. Both the miRNAs directly inhibited cyclin T2 gene, preventing the release of hypophosphorylated retinoblastoma and resulting in induction of monocytic differentiation. In addition, a target of miR-29a, cyclin-dependent kinase 6 gene, and a target of miR-142-3p, TGF-ß-activated kinase 1/MAP3K7 binding protein 2 gene, are involved in the regulation of both monocytic and granulocytic differentiation. A significant decrease of miR-29a and 142-3p levels and an obvious increase in their target protein levels were also observed in blasts from acute myeloid leukemia. By lentivirus-mediated gene transfer, we demonstrated that enforced expression of either miR-29a or miR-142-3p in hematopoietic stem/progenitor cells from healthy controls and acute myeloid leukemia patients down-regulated expression of their targets and promoted myeloid differentiation. These findings confirm that miR-29a and miR-142-3p are key regulators of normal myeloid differentiation and their reduced expression is involved in acute myeloid leukemia development.

Treatment and complications of thalassemia major in Guangxi, Southern China.

BACKGROUND: ß-Thalassemia is extremely prevalent in Guangxi province, Southern China. However, little is known about the treatment and complications of patients with thalassemia major (TM) in Guangxi. The first thalassemia center in China was opened in Guangxi in 2003. Since that time, more than 400 patients have been enrolled. PROCEDURE: From December 2009 to February 2010, data was collected from TM patients visiting the thalassemia center including the circumstances of diagnosis, biological and clinical data, markers of iron overload and treatment. RESULTS: Data on 231 patients (median age, 5 years; range, 5 months to 21 years) were recorded. Only 44.6% of patients maintained their hemoglobin levels >9.0 g/dl. In 186 patients with ferritin levels >1,000 ng/ml, an iron chelator was used regularly in 44.6%, irregularly in 26.9%, and was not used in 28.5%. The mean serum ferritin level was 3,143 ng/ml and levels increased with age. Height and weight retardation were found in 48.3% and 11.1% patients, respectively. Compared to patients treated outside of the center, patients completing treatment in the thalassemia center had a higher hemoglobin level before transfusion, higher height and weight SD score, and less splenomegaly, but a similar ratio of regular or irregular iron chelation. Six (18.2%) of 33 patients >10 years of age (14.3 ± 2.8 years; range, 11-19 years) were diagnosed as hypothyroid. CONCLUSIONS: Although survival status of patients with TM in Guangxi has improved since the opening of the thalassemia center, TM complications remain high and with an early onset.

A case report of concomitant paroxysmal nocturnal hemoglobinuria and heterozygous ß-thalassemia.

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonal blood disorder that presents chronic intravascular hemolysis. PNH concomitant with inherited hemolytic anemia has been rarely reported. Here, we report an interesting PNH patient who was misdiagnosed with iron deficiency anemia due to concomitant heterozygous ß-thalassemia. The patient experienced dizziness, fatigue, and restricted physical activity for the previous 3 years. Thalassemia gene analysis revealed heterozygous ß-thalassemia. Iron staining of the bone marrow demonstrated the absence of stainable iron and sideroblasts. The patient was diagnosed with iron deficiency anemia. Iron supplementation treatment was performed, but the anemia remained unresolved. The patient became transfusion dependent 1 year later and was admitted to our hospital in March 2010. Flow cytometry of the patient's peripheral blood demonstrated that 7.9% and 11.9% of the erythrocytes were CD59 and CD55 deficient, respectively. The patient was finally diagnosed with concomitant PNH and heterozygous ß-thalassemia.

Refractory platelet transfusion in a patient with CD36 deficiency due to pseudothrombocytopenia.

Type I CD36 deficiency is defined by the absence of CD36 on both platelets and monocytes. Pseudothrombocytopenia (PTCP) is characterized by a false reduction in the number of platelets in ethylenediaminetetraacetic acid (EDTA)-anticoagulated blood. Here we report a rare case of concomitant CD36 deficiency and PTCP. The patient was a 7-year-old boy who suffered comminuted fractures of the left humeral condyle. In the pre-operative examination, he was found to have thrombopenia and assumed to have idiopathic thrombocytopenic purpura. After immunotherapy and platelet transfusion, the platelet count remained low, suggesting that the patient was refractory to platelet transfusion. Serum was collected for the detection of platelet antibodies, and antibodies against CD36 were found. Flow cytometry verified the absence of CD36 on both the platelets and monocytes of this patient. However, the platelet count was normal when capillary blood smears were analysed; in addition, platelet coagulation was noted under the microscope when EDTA-anticoagulated peripheral blood was used. The patient underwent surgery without platelet transfusion and recovered uneventfully.

[Intracerebral infiltration by monoclonal plasmacytoid cells in Waldenstrom's macroglobulinemia: a case report and review of the literature].

OBJECTIVE: To sum up the clinical experience of the diagnosis and treatment of intracerebral infiltration by monoclonal plasmacytoid cells in Waldenstrom's macroglobulinemia(Bing-Neel syndrome). METHODS: The clinical data of the diagnosis and treatment of a case of Bing-Neel syndrome was analyzed. RESULTS: A 56-year-old male was diagnosed as Waldenstrom's macroglobulinemia one year ago, and presented with persistent headache during the treatment period. Magnetic resonance imaging showed a high intensity area on T2-weighed images in the right frontal lobe which was well enhanced by gadolinium-diethylenetriaminepenta-acetic acid. Infiltration of neoplastic cells was confirmed by biopsy. Immunohistochemical examination showed that mature plasmacytoid cells in the cerebral parenchyma were immunoglobulin M positive. CONCLUSION: Infiltration in CNS (Bing-Neel syndrome) is uncommon in Waldenstrom's macroglobulinemia. As there is no effective therapy for this Bing-Neel syndrome, combination of radiation and chemotherapy should be considered for this situation.

[Relationship between the genetic background of donor KIR recipient HLA and the outcomes in HLA-identical sibling HSCT].

OBJECTIVE: To explore the relationship between the genetic background of donor KIR/recipient HLA and the outcomes in HLA-identical sibling HSCT. METHODS: HLA genotype was determined by polymerase chain reaction-sequence-specific oligonucleotide probes (PCR-SSOP) and/or PCR-sequence-specific primer (PCR-SSP). Donor KIR genotype was determined by PCR-SSP. A retrospective study was carried out to analyze the outcomes of 59 patients with various hematologic malignancies received non T-cell-depleted transplant from HLA-identical sibling donors. RESULTS: Incidence of grade II-IV acute graft-versus-host disease (aGVHD) was significantly lower in patients of KIR/HLA matched group than in KIR/HLA mismatched group (32% vs 78%, P = 0.026). The incidence of grade II-IV aGVHD (24% vs 61%, P = 0.018) and fungus infection (14% vs 44%, P = 0.028) were significantly lower in Bw4 matched group than in Bw4 mismatched group. In myeloid diseases, Bw4 matched patients had much lower incidence of fungus infection (12% vs 80%, P = 0.002) compared with Bw4 mismatched patients, and C2 matched patients had higher overall survival (OS) compared with C2 mismatched patients (P = 0.01). CONCLUSIONS: Donor KIR/recipient HLA genetic background is correlated with the outcomes of HLA-identical sibling HSCT in incidences of grade II-IV aGVHD, fungus infection and OS. KIR/HLA matched patients may have lower incidence of aGVHD. Bw4 matched patients may have lower incidences of aGVHD and fungus infection. C2 matched patients may have longer OS.