Clinical efficacy of CCG7942/POG9354 protocol in treatment of peripheral primitive neuroectodermal tumor in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical manifestations, diagnosis, and treatment of peripheral primitive neuroectodermal tumor (pPNET) in children and the survival of patients treated with the CCG7942/POG9354 protocol.</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 10 patients with pPNET admitted from October 2008 to October 2013. Of the 10 patients, 3 had metastasis, while others had no metastasis. The 7 patients without metastasis were treated with the Children's Cancer Study Group 7942 (CCG7942) protocol, and the other 3 patients with metastasis with the Pediatric Oncology Group 9354 (POG9354) protocol. The therapeutic response and chemotherapy-related toxicities were evaluated by WHO criteria and Common Terminology Criteria for Adverse Events (version 4.0).</p><p><b>RESULTS</b>In the 7 patients treated with the CCG7942 protocol, 4 achieved a complete remission (CR), 1 had stable disease, 2 developed progressive disease (PD), and 2 had recurrence. In the 3 patients treated with the POG9354 protocol, 1 achieved a CR, 2 developed PD, 2 had recurrence, and 2 died. For the 7 patients without metastasis, the survival time was 5-60 months, and the event-free survival rate was 71%. For the 3 patients with metastasis, the survival time was 13-25 months, and the event-free survival rate was 33%. All patients developed grade 4 bone marrow suppression, and other grade 1-2 toxicities, including gastrointestinal reactions, liver function impairment, and renal function impairment, were also seen.</p><p><b>CONCLUSIONS</b>CCG7942 protocol is effective and safe for children with non-metastatic pPNET. However, POG9354 protocol has unsatisfactory efficacy in children with metastatic pPNET, so further studies are needed to improve the therapy for this disease.</p>

A review on treatment of high-risk neuroblastoma / 中国当代儿科杂志

So far treatment of advanced neuroblastoma is still difficult, due to its high malignancy. Currently comprehensive therapies, including high-dose multi-drug chemotherapy, surgery, stem cell transplantation, radiation, biological therapy and immune therapy as well as target therapy dominant the treatment of this disease, and we hereby introduce the latest development of treatment protocols for this disease.

Treatment of endodermal sinus tumor in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the treatment and outcome of childhood endodermal sinus tumor.</p><p><b>METHODS</b>The clinical data of twelve children with endodermal sinus tumor between April 2000 and July 2013 were reviewed. The basic demographics, stages of the lesion and the treatment outcome were analyzed. Of the twelve patients, seven were boys and five were girls. The age of the disease onset was between 1 and 3.3 years, except one in 11 years. Two patients were in Brodeur Stage I, four in Stage II, two in Stage III, and four in Stage IV. One patient underwent surgery alone, one underwent surgery plus a combination therapy with vincristine, actinomycin and cyclophosphamide (VAC), and the other ten were treated by surgery with the use of cisplatin, etoposide and bleomycin (PEB) before or after the operation.</p><p><b>RESULTS</b>Eleven patients were successfully followed up and ten were alive. The length of survival was 4.5 to 66 months in the 10 patients. In the 10 patients treated with PEB before or after surgery, 8 achieved complete remission, one achieved partial remission and one was not followed up. The major complications associated with the PEB regimen included myelosuppression and gastrointestinal upset symptoms and no late toxicity was observed.</p><p><b>CONCLUSIONS</b>Preoperative or postoperative administration of PEB may be an effective and safe management modality for childhood endodermal sinus tumor. Nevertheless, further validation is warranted in prospective studies involving a larger sample size.</p>

Haploidentical hematopoietic stem cell transplantation for the treatment of severe aplastic anemia in children / 中国实验血液学杂志

This study was purposed to assess the effectiveness of haploidentical hematopoietic stem cell transplantation (HSCT) without in vitro T cell depletion for the treatment of severe aplastic anemia (SAA) in children. Two children with SAA/very SAA (VSAA) received T cell-depleted HSCT from their fathers with 2 loci mismatched in our center between October 2010 and March 2013. During 4 months after onset, both failed in treatment of cyclosporine (CsA)+ granulocyte colony stimulating factor (G-CSF), had active or serious infections, were transfusion dependent and lacked HLA-identical sibling donors and unrelated donors. The conditioning regimen before HSCT included fludarabine, cyclophosphamide and thymoglobulin. The source of grafts was a combination of G-CSF mobilized peripheral blood hematopoietic stem cells and BM. The recipients received CsA, mycophenolate mofetil (MMF) and short-term MTX for GVHD prophylaxis. Both children with SAA achieved 100% donor myeloid engraftment. Neutrophil engraftment occurred at day 12 and day 18 after transplant respectively. Platelet engraftment occurred at day 17 and day 26 after transplantation respectively. Two patients all developed grade I acute graft versus host disease (GVHD), one of which evolved into chronic limited GVHD well-controlled. Both have survived for two years after transplantation with 100% donor myeloid engraftment and effective lymphoid reconstitution. In conclusion, these limited cases suggest that haploidentical HSCT for children with SAA without a HLA-identical sibling donor and unrelated donor may be feasible. Further prospective clinical study is required to increase the overall survival (OS) by decreasing GVHD while maintaining stable engraftment.

Treatment of nasopharyngeal carcinoma in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy of the ARAR0331 protocol for treatment of childhood nasopharyngeal carcinoma.</p><p><b>METHODS</b>The clinical data of eight children with nasopharyngeal carcinoma between May 2004 and May 2012 were retrospectively studied. The eight patients included six boys and two girls, and the onset age was between 3 and 13 years. Six patients were in AJCC Stage Ⅲ, one was in StageⅡA and one was in Stage ⅣA. One patient had been treated with combined radiotherapy and chemotherapy which mainly included EAP, BEP and EA. The other seven patients had been treated with the ARAR0331 protocol provided by the America Children's Oncology Group (COG).</p><p><b>RESULTS</b>The patient who had been treated with combined radiotherapy and chemotherapy developed multiple bony metastasis during the chemotherapeutic period. Four out of seven patients who had been treated with ARAR0331 protocol achieved complete remission, and two achieved partial remission. The seven patients were followed-up from 8 to 75 months and the survival rate was 100%. The ARAR0331 protocol treatment-related complications included radiodermatitis, mucocitis and nausea. Late toxicity was not found.</p><p><b>CONCLUSIONS</b>Based on the limited cases, ARAR0331 protocol appears to be effective and safe for childhood nasopharyngeal carcinoma.</p>

Effects of knocking down Foxp3 on immunological functions of human neuroblastoma SK-N-BE2 cell / 中华实用儿科临床杂志

Objective To investigate whether knockingdown Foxp3 affects the immunological function of neuroblastoma cells.Methods Different siRNAs of Foxp3 were designed and the one that effectively inhibited Foxp3 gene expression was selected by real-time polymerase chain reaction and by fluorescence activated cell sorter analysis.Effects of Foxp3 knocking down on the expression of co-stimulatory molecule CD86 were also examined.Finally,neuroblastoma cells silenced of Foxp3 expression were co-cultured with human peripheral blood monouclear cells and the activities of T cells were examined.Results Expression of Foxp3 in SK-N-BE2 cells could be efficiently knocking down in both mRNA and protein levels.Knocking down Foxp3 increased the expression of CD86 and moderately increased IFN-γand IL-17 expressions,and significantly inhibited the expressions of TGF-β and IL-10.Conclusion Silencing Foxp3 expression in neuroblastoma SK-N-BE2 cells reverses its immune evasion mechanisms,which suggests that this is a new strategy for treating this kind of diseases.

Childhood rhabdomyosarcoma: a retrospective review of 23 cases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics, treatment and outcome of childhood rhabdomyosarcoma.</p><p><b>METHODS</b>The clinical data of 23 children with rhabdomyosarcoma from January, 1998 to October, 2008 were retrospectively reviewed.</p><p><b>RESULTS</b>Of the 23 cases, 15 were male and 8 were female, with a mean age of 5 years old (7 months to 12 years old). Based on the American IRS staging system, 2 cases were in stage I, 4 cases in stage II, 8 cases in stage III, and 9 were in stage IV. The primary sites were found in head and neck (14 cases), extremities (4 cases), bladder (2 cases), kidney (1 case), post-peritoneum (1 case) and bile duct (1 case). All of the children were confirmed with rhabdomyosarcoma by biopsy and immunohistochemistry. The clinical manifestations were related to the tumor tissues-induced space occupying, compression and erosion and were aspecific. The patients in different IRS stages were given different treatment regimens. The chemotherapy regimens VDCA, VAC or VadrC were used before 2002. After 2002, the Children's Oncology Group (COG) protocol was employed. The two-year survival rate was 63% in 19 patients who received a combination of surgery, chemotherapy and radiotherapy, but none of 4 patients who received a surgery alone or a combination of surgery and chemotherapy or radiotherapy survived more than two years.</p><p><b>CONCLUSIONS</b>The clinical manifestations of childhood rhabdomyosarcoma are not specific. A combination therapy including surgery, chemotherapy and radiation is effective to the improvement of the survival rate in children with rhabdomyosarcoma.</p>

Neuroblastoma LA-N-6 cells express Foxp3 which can be suppressed by chemotherapeutic agents / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate whether neuroblastoma cells LA-N-6 express Foxp3 and whether the expression of Foxp3 is sensitive to chemotherapy by cyclophosvnamide (CTX)and pirarubicin (THP).</p><p><b>METHODS</b>Expression of Foxp3 on LA-N-6 cells was examined by flow cytometry analysis. The dose-effects of chemotherapy drugs including CTX and THP on LA-N-6 cells were investigated by MTT assay. The effects of CTX and THP on Foxp3 expression were examined by flow cytometry and real-time PCR assays.</p><p><b>RESULTS</b>Flow cytometry analysis showed that LA-N-6 cells expressed Foxp3 at a high level. At sub-optimal concentration, chemotherapy drugs CTX and THP significantly down-regulated expression of Foxp3 on LA-N-6 cells at protein level (P<0.05). CTX also decreased the expression of Foxp3 at mRNA level (P<0.05). CONCLSUSIONS: Neuroblastoma cells LA-N-6 express Foxp3 at a high level, which can be suppressed by chemotherapy drugs CTX and THP. These data suggest that chemotherapy might suppress the growth and metastasis of tumor cells partially through inhibiting the expression of Foxp3.</p>

Effect of topotecan on MAGE gene expression of HPB-AM cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate whether topotecan, a novel anti-tumor agent, down-regulates gene expression of melanoma antigen-encoding (MAGE) in HPB-AM cells.</p><p><b>METHODS</b>MAGE mRNA expression of HPB-AM cells was detected by RT-PCR 4, 8, 12 and 16 hrs after different concentrations (0.05, 0.10, 0.15 and 0.20 micromol/L) of topotecan treatment.</p><p><b>RESULTS</b>MAGE mRNA expression of HPB-AM cells decreased with increasing concentrations of topotecan 12 hrs after treatment. The MAGE mRNA expression of HPB-AM cells treated by 0.10, 0.15 and 0.20 micromol/L of topotecan was significantly lower than that in the blank control group (P<0.05). MAGE mRNA expression of HPB-AM cells was significantly reduced in a time-dependent manner after 0.10 micromol/L of topotecan treatment. The MAGE mRNA expression of HPB-AM cells treated by 0.10 micromol/L of topotecan was significantly lower than that in the blank control group 12 and 16 hrs after treatment (P<0.05).</p><p><b>CONCLUSIONS</b>Topotecan is capable of inhibiting the expression of MAGE mRNA of HPB-AM cells in a time- and dose-dependent manner.</p>

Curative effects of the protocol of CDV combined with CiE as pre-operative chemotherapy in high-risk childhood neuroblastoma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the effects and the toxicity of the protocol of CDV combined with CiE as pre-operative chemotherapy in childhood stage IV neuroblastoma.</p><p><b>METHODS</b>The clinical data of 27 children aged from 1.2 to 8 years with neuroblastoma in stage IV was retrospectively studied. The primary sites of the diseases were abdomen (n = 21), posterior mediastinum (n = 4) and pelvic cavity (n = 2). Twenty three patients had bone marrow metastasis. Twelve patients had bone metastasis. All patients were treated with the CDV protocol (cyclophosphamide + doxorubicin + vincristine) for 3 cycles and the CiE protocol (cisplatin + etoposide) for 2 cycles. Neuroblastoma therapeutic response evaluation criterion and common terminology criteria for adverse events of National Cancer Institute were used to evaluate effects and chemotherapy related toxicity.</p><p><b>RESULTS</b>All patients received the pre-operative chemotherapy. The overall response rate was 82%. After chemotherapy, 24 patients received operations. Total resection of primary tumor was found in 14 patients (58%) and part resection in 10 patients (42%). The most common chemotherapy related toxicity was bone marrow suppression: grade IV suppression of neutrophils (n = 27), reduction in hemoglobin (III grade, n = 7; IV grade, n = 20) and reduction in platelet (III grade, n = 2; IV grade, n = 25). Infection was found in all patients and was controlled with antibiotics. I or II grade lesions of digestive, liver and kidney were found and could be recovered after therapy. Grade I neurotoxicity occurred in 2 patients (7%). The heart function damage was not found in any of patients.</p><p><b>CONCLUSIONS</b>The protocol of CDV combined with CiE as pre-operative chemotherapy might be effective in children with stage IV neuroblastoma.</p>

Treatment of acute leukemia complicated by invasive aspergillosis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the antifungal treatment and intensive chemotherapy in children with acute leukemia and invasive aspergillosis.</p><p><b>METHODS</b>The diagnosis and treatment of 4 cases of childhood acute leukemia complicated by invasive aspergillosis between July 2007 and July 2008 were studied retrospectively.</p><p><b>RESULTS</b>Three children who underwent remission induction chemotherapy for ALL and one who underwent consolidation chemotherapy for AML developed invasive aspergillosis. One child with proven aspergillosis and 3 with possible aspergillosis all had halo sign on CT at diagnosis. Voriconazole or amphotericin B was given as primary therapy. Improvements of fungal lesions were shown by CT after two to four weeks of antifungal therapy. Complete radiologic remissions were achieved between 4 months and one year. The intensive chemotherapy schedule was continued in all of 4 cases. The median time from fungal infection to the continuation of chemotherapy was 35 days. None showed recurrence of fungal infection.</p><p><b>CONCLUSIONS</b>The halo sign on CT may be a reliable indicator for the early diagnosis of invasive aspergillosis. The preemptive antifungal therapy on the basis of the identification of a halo sign and the reversal of immunosuppression may improve the outcome of invasive aspergillosis. Prolonged antifungal treatment during subsequent cycles of chemotherapy permits completion of scheduled intensive chemotherapy without fungal recurrence.</p>

Response to therapy of 13 children with rhabdomyosarcoma / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical response to comprehensive therapy in children with rhabdomyosarcoma.</p><p><b>METHODS</b>Clinical data of 13 children (8 males and 5 females) with rhabdomyosarcoma from January 1998 through October 2005 were retrospectively studied. Their ages ranged from 7 months to 12 years. The 13 cases of rhabdomyosarcoma consisted of 2 cases in stage I, 2 cases in stage II, 3 cases in stage III, and 6 cases in stage IV. Rhabdomyosarcoma was confirmed by biopsy, 12 cases (92.3%) presenting as embryonal type and 1 as alveolar type in histology. One patient underwent surgery treatment alone, one patient received surgery plus local radiation treatment, one patient received surgery plus chemotherapy and 10 patients were administered with a combination of surgery, local radiation treatment and chemotherapy. The chemotherapy protocol before 2002 was VDCA, VAC or VadrC. After 2002, the COG protocol was employed, with CDV+IE for stage III, and CT+VAC or CT+VAC+VCT for stage IV patients.</p><p><b>RESULTS</b>The 2-year overall survival was 60% in the 10 patients who received a combination of surgery, local radiation treatment and chemotherapy, but the three patients died without receiving combination therapy. The 2-year overall survival in the 13 patients was 46.2%. The 2-year overall survival of the patients after 2002 (60%, 3/5) was higher than that before 2002 (37.5%, 3/8).</p><p><b>CONCLUSIONS</b>Embryonal rhabdomyosarcoma dominates the histology type in children, which is highly malignant. A combination therapy of surgery, local radiation and chemotherapy can result in a satisfactory therapeutic effect in children with rhabdomyosarcoma.</p>

Characteristics of chronic active Epstein-Barr virus infection-associated hematological disorders in children / 中国实验血液学杂志

The aim of this study was to analyze characteristics of chronic active Epstein-Barr virus (CAEBV) infection associated hematological disorders in children. Clinical characteristics were summarized; the morphology of hematopoietic cells in bone marrow was observed by microscopy; the lymphocyte subpopulations were analyzed by flow cytometry; the immunophenotype of liver biopsies was assayed by immunohistochemistry; EBV-related antibodies were measured by ELISA; serum EBV-DNA loads were detected by real-time quantitative PCR; EBV-encoded small RNA 1-positive cells in peripheral blood mononuclear cells were identified by in situ hybridization. The results indicated that the clinical manifestations in patients included persistent or recurrent fever, hepatosplenomegaly, liver dysfunction, anemia, thrombocytopenia, systemic inflammatory reaction. Bone marrow presented as hypocellularity, dysmaturation, myelodysplasia and hemophagocytosis. CD8(+) cell high counts were demonstrated in all 4 patients, one of them developed into a T cell lymphoma. Serum EBV-DNA load was 3.26 x 10(3) copies/ml in one patient, EBER1(+) cells were detected at a frequency of 1.7% in PBMNCs from another patient; the titers of IgG to EBV-VCA were >or= 1:5120 in the rest 2 patients. All 4 patients described above were diagnosed as CAEBV infection. In conclusion, the immune-related cytopenia, macrophage activation syndrome and lymphoproliferative disorders are characteristics of CAEBV infection associated hematological disorders in these 4 children patients.

Matrine inhibits the proliferation of neuroblastoma LA-N-5 cell and MYCN mRNA expression / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Neuroblastoma is the most common malignant solid tumor in children under 4 years. Amplification of MYCN oncogene is associated with advanced-stage disease, rapid tumor progression, resistance to treatment, and poor outcome. Matirne has the anti-tumor activity. This study was designed to investigate the effects of matrine on LA-N-5 cell line proliferation and MYCN gene mRNA expression.</p><p><b>METHODS</b>Neuroblastoma LA-N-5 cells were treated by 0.25, 0.50, 0.75 or 1.00 mg/mL matrine. MTT was used to measure the levels of the proliferation of LA-N-5 cells cultured with different concentrations of matrine. MYCN gene mRNA expression in LA-N-5 cells was measured using real time RT-PCR with SYBR GREEN I fluorescence.</p><p><b>RESULTS</b>The proliferation of LA-N-5 cells was obviously inhibited by matrine in a dose- and time- dependent manner. Matrine of 1.00 mg/mL treatment for 72 hrs produced a best effect, with an inhibitory rate of LA-N-5 cell proliferation of 36.3% and an inhibitory rate of MYCN gene mRNA expression of 44.6%.</p><p><b>CONCLUSIONS</b>Matrine may inhibit the growth of neuroblastoma cells and down-regulate MYCN mRNA expression. It may be promising as a new drug for treatment of neuroblastoma.</p>

Childhood acute megakaryoblastic leukemia / 中国实验血液学杂志

The aim of this study was to investigate the clinical, pathological and biological features of acute megakaryoblastic leukemia in childhood. The morphology of cells was observed by means of bone marrow smear; the immunophenotype was detected by flow cytometry and immunohistochemistry assay. The results indicated that the fever, hemorrhage, hepatosplenomegaly and lymphadenopathy in this case were the primary presentations accompanying by leukocytosis, anemia and thrombocytopenia. An adequate marrow aspirate could not be obtained. At the time of diagnosis, the bone marrow had more than 30% megakaryoblasts in nucleated cells. Flow cytometric analysis revealed the dual expression of CD41 and CD61 by tumor cells in bone marrow. The histopathological examination of bone marrow demonstrated infiltration of large-sized CD42b(+) cells. According to all above mentioned results, this case was diagnosed as acute megakaryoblastic leukemia. In conclusion, childhood acute megakaryoblastic leukemia is a rare and easily misdiagnosed disease with poor prognosis. Flow cytometry analysis and immunohistochemistry assay of bone marrow can help in detecting this leukemia subtype and evaluating its prognosis.

Skin lesions and myelodysplastic syndrome as initial manifestations of biphenotypic acute leukemia / 中国实验血液学杂志

The aim of this study was to investigate the clinical, pathological and biological features of biphenotypic acute leukemia. The morphology of tumor cells was observed by bone marrow examination; the immunophenotype was assayed by flow cytometry and immunohistochemistry; the chromosomal aberrations were detected by conventional chromosomal analysis and RT-multiplex nested PCR. The results showed that extramedullary skin lesions and myelodysplasia occurred before the onset of overt disease. At the time of diagnosis, this case had more than 30% blasts in bone marrow with meningeal involvement. Large-sized tumor cells predominated morphologically over other cells. Flow cytometry revealed the co-expression of myeloid antigens (cMPO, CD33 and CD117) and T-lymphoid antigens (cCD3, CD5, CD7, dual expression of CD4 and CD8). Immunohistochemical staining showed that CD43 and CD99 were strong positive which define the earliest hematopoietic progenitors. Partial tandem duplication of the MLL gene could be detected with normal cytogenetic method. All above-mentioned results led to the diagnosis of biphenotypic acute leukemia. It is concluded that the biphenotypic acute leukemia is an uncommon type of leukemia which may be preceded by myelodysplastic syndrome and has aggressive clinical and biological behavior. Immunophenotype, cytogenetics and molecular analysis can contribute to early diagnosis of BAL and evaluation of prognosis.

Ex vivo chemical modification of graft cells by methoxy polyethylene glycol alleviates graft versus host disease after haploidentical stem cell transplantation in mice / 中国实验血液学杂志

This study was aimed to explore the effect of ex vivo chemical modification of graft cells with methoxy polyethylene glycol (mPEG) on graft versus host disease (GVHD) after haploidentical stem cell transplantation in neonatal mice and its influence on activity of the stem cells. The modified and non-modified spleen cells of adult CB6F1 mice were injected into the abdominal cavity of neonatal BALB/c mice with 5x10(6) spleen cells per mouse, and GVHD were measured by spleen index (SI). Furthermore, the modified and non-modified mixture of bone marrow and spleen cells (BMS) were transplanted to haploidentical lethally irradiated adult BALB/c mice via tail vein with 2x10(5) BMS per mouse, and the colony forming units of spleens (CFU-S) were counted on the eighth day after irradiation. The results indicated that SI1 in modification group were lower than that in non-modification group, and SI2 in modification group was <1.3, showing that GVHD in modification group were less severe. The numbers of CFU-S formed in both modification group and non-modification group were not significantly different (p>0.05), indicating that the activity of the stem cells were not affected by mPEG modification. In conclusion, the modification of graft cells with mPEG alleviates GVHD after haploidentical stem cell transplantation in neonatal mice, and do not influence the activity of the stem cells.

Clinical features and prognosis of advanced neuroblastoma in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the clinical features, treatment modalities and the prognosis of advanced neuroblastoma in children.</p><p><b>METHODS</b>The medical records of 63 children with stage III or IV neuroblastoma from January 1996 to December 2005 were retrospectively reviewed. Sixty patients were treated by tumor resection and (or) chemotherapy and (or) radiation. Fourteen out of the 60 patients received another autologous peripheral blood stem cell transplantation.</p><p><b>RESULTS</b>Of the 63 patients with advanced neuroblastoma, the male/female ratio was 2.7:1 and the median age at diagnosis was 4 years old. Most of the initial symptoms included pyrexia, abdominal pain, abdominal mass, and leg or articular pain. Primary tumor sites were adrenal (38%), retroperitoneal (35%), mediastinal (17%), pelvic (6%) and cervical (2%). The sites of metastasis at diagnosis included local (41%) and (or) distant (37%) lymph nodes, bone marrow (60%), bone (46%) and liver (16%). The median survival time of the 63 patients was 32.7 months. The 2-year survival rate was 44.3%. Statistical analysis demonstrated that unfavorable survival prognostic factors were the following: age > 1 year at diagnosis (P < 0.05); serum neuro-specific enolase > 100 mg/L (P < 0.05); serum lactic dehydrogenase > 1500 U/L (P < 0.01); serum ferritin >150 mg/L (P < 0.05). The overall survival period of the patients was prolonged through total resection of the primary tumor (P < 0.05). Intensive chemotherapy in combination with autologous peripheral blood stem cell transplantation could also result in a prolonged overall survival period (P < 0.01).</p><p><b>CONCLUSIONS</b>Neuroblastoma with advanced stages often presents with various clinical manifestations and has a poor prognosis. It is beneficial to improve the prognosis of neuroblastoma through an early diagnosis and a comprehensive therapy including total resection of the primary tumor, autologous peripheral blood stem cell transplantation and intensive chemotherapy.</p>

Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To examine the feasibility and practicability of quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) with SYBR GREEN I fluorescence for detecting the MYCN mRNA expression in neuroblastoma cell line LA-N-5.</p><p><b>METHODS</b>MYCN mRNA expression in LA-N-5 cells was measured using real time RT-PCR with SYBR GREEN I. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as internal control. The level of the MYCN mRNA was calculated as MYCN copies/GAPDH copies.</p><p><b>RESULTS</b>Standard curves were linear and showed high correlations (R2>0.99). The ratio of MYCN mRNA copies to GAPDH mRNA copies was calculated based on specific PCR products. The MYCN mRNA level in LA-N-5 cells was obtained (17.4 +/- 1.2).</p><p><b>CONCLUSIONS</b>Quantitative RT-PCR with SYBR GREEN I fluorescence may be a sensitive and reliable method for detecting the MYCN mRNA expression. It may also be potential applicable for detecting the MYCN mRNA expression in the small amount neuroblastoma tissues.</p>

Relevant low toxicities with rhG-CSF mobilized and cryopreserved autologous peripheral blood stem cell return infusions in children / 中国实验血液学杂志

The purpose of this study was to evaluate the safety of cryopreserved and thawed peripheral blood stem cell (PBSC) fractionated return infusions in children. 35 children patients with malignant tumors (13 acute leukaemias, 15 neuroblastomas and 7 malignant lymphomas) received fractionated return infusions of cryopreserved stem cells after undergoing high-dose chemotherapy without or with total body irradiation. The toxicities of 70 return infusions were evaluated. All patients were mobilized by chemotherapy plus recombination human granulocyte colony-stimulating factor (rhG-CSF), and then PBSCs were collected by a separator CS-3000 plus or COBE spectra-4. The grafts were cryopreserved in 10% dimethyl sulfoxide (DMSD) and stored in liquid nitrogen. There were totally 70 PBSC transfusions. The total volume of PBSCs transfused: 190 - 420 ml (265 +/- 73 ml or 13.7 +/- 4.2 ml/kg) with a mean of (4.43 +/- 1.91) x 10(8)/kg of PBSCs, and 0.94 +/- 0.18 g/kg of DMSO. The single dose: 90 - 300 ml (132 +/- 37 ml or 6.6 +/- 5.2 ml/kg) with a mean of 0.68 +/- 0.12 g/kg of DMSO. Symptoms occurring during the infusions were recorded. All patients were monitored for 24 hours after infusion. Pulse, blood pressure, body temperature, and respiratory rate were recorded every 15 minutes. At four hours before and 8 hours after infusion, urinalysis was performed. Serum potassium, sodium, creatinine, total bilirubin, aspartate amino transferase (AST), and alanine amino transferase (ALT) levels were examined within 24 hours before and after the first infusion. The results showed that the toxicities observed included hemoglobinuria in 54 return infusions (77.1%), headache in 28 (40.0%), nausea in 24 (34.3%), vomiting in 17 (24.3%), and abdominal pain in 8 (11.4%). Patients who received a graft > 200 ml tended to have a higher frequency of hemoglobinuria, headache, nausea, vomiting, or abdominal pain (P<0.01), and they disappeared quickly, too. Total bilirubin increased after the first return infusion (P<0.01), and there was a significant correlation between the volume of infusion and the degree of total bilirubin increase (r=0.8977, P<0.01). No renal failure or shock occurred. It is concluded that transient hemoglobinuria, headache, nausea, vomiting, and abdominal pain are common toxicities associated with PBSC autograft, and these toxicities are related with a single volume of PBSCs transfused. Total bilirubin increase is correlated with the volume of infusion. In a word, the toxicity is less frequent and lower severe in children with fractionated infusions of cryopreserved peripheral blood stem cell.