OBJECTIVETo investigate the expression of CD19 on childhood acute leukemia (AL) and its significance, and to provide evidence for the diagnosis and differential diagnosis as well as monoclonal antibody-targeting treatment of leukemia.

METHODSThere were 210 cases of childhood AL, of which 130 cases were male and 80 were female with a mean age of 9 years old. Using a panel of 27 fluorochrome directly labeled monoclonal antibodies, 210 samples from the patients were analyzed with CD45/SSC double parameters and multi-color flow cytometry to determine the expression of CD19.

RESULTSIn the 93 cases of B lineage acute lymphoblastic leukemia (ALL), the positive rate (98.9%, 92/93) of CD19 was significantly higher than that of the other B cell related antigens, such as CD10 (88.2%, 82/93, P = 0.003), CD20 (24.7%, 23/93, P = 0.001) and CD22 (60.2%, 56/93, P = 0.001). CD19 was expressed on all 8 cases of B/myeloid (My) hybrid acute leukemia (HAL) and 1 case of B/T HAL, but was not expressed on all 24 cases of T lineage leukemia and 5 cases of T/My HAL. In the 79 cases of acute myeloid leukemia (AML), only 5 (6.3%) cases expressed CD19. The positive rate (6.3%) of CD19 on AML was significantly lower than that on B lineage ALL (98.9%, P = 0.001). The percentage of CD19 positive cells in B/My HAL (41.6% - 88.7% with a mean of 73.8%) was significant higher than that in CD19(+)-AML (21.4% - 50.4% with a mean of 24.2%; Run Sum test, P = 0.0084). Of the 210 cases, 102 were B lineage related AL including B lineage ALL, B/My HAL and B/T HAL. In B lineage related AL, the sensitivity and the specificity of CD19 was 99.0% (101/102) and 95.4% (13/108) while the positive predictive and the negative predictive values to B lineage were 95.3% (101/106) and 99.0% (103/104), respectively. Using CD19(+) as a single reagent to diagnose B lineage, the false positive rate was 4.6% (5/108) and the false negative rate was 1.0% (1/102) with a general diagnosis index (GDI) of 94.4% [GDI = 1-(false positive rate + false negative rate)].

CONCLUSIONCD19 is continuously and stably expressed on all stages of B lineage differentiation. It is a reliable cell membrane marker for diagnosing B lineage ALL and an ideal target for antibody-targeting treatment of leukemia as well; the expression degree of CD19 can be used to distinguish B/My HAL from CD19(+)-AML; CD19 didn't express on normal myeloid cells but did on some AML cells. Therefore it can be used to detect the minimal residual disease.

Adolescent ; Antigens, CD19 ; analysis ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology

OBJECTIVETo observe the incidence of elimination delay in high dose methotrexate (HDMTX) therapy, its side effects and influence to next course of chemotherapy and analyze the relationship between the dosage, the duration of MTX infusion and the morbidity of the elimination delay.

METHODSA total of 121 childhood acute lymphoblastic leukemia (ALL) (497 infusions of HDMTX) were analysed in this study. The elimination delay rate and the adverse effects in different dose groups (3 g/m2 vs 5 g/m2) and different infusion duration groups (7 h vs 24 h) were compared. The adverse effect evaluation was based on the World Health Organization (WHO) Toxicity Grading Criteria. The rescue dosages of calcium folinate (CF) among these groups were compared through CF/MTX index.

RESULTSThe overall morbidity of elimination delay was 12.1% with a relative risk of 30.6% for the first time. The relative risk for the second time of occurrence was increased to 45.9% (P < 0.01) and it was not significantly increased for the third time (35.3%). Children with elimination delay had lower platelet count (P < 0.01) and higher CF rescue dosage (P < 0.01), while the damage of oral mucous membrane was more severe (P < 0.05) and the next course of chemotherapy would be postponed for a median of 4 days in 3 g group. There was no significant difference in elimination delay rates between 3 g and 5 g groups (12.1% vs 12.0%, P > 0.05), and between 7 h and 24 h MTX infusion groups (13.6% vs 11.9%, P > 0.05). The only side effect occurred in 5 g group was gastrointestinal morbidity. The CF/MTX index of 5 g group without elimination delay was less than that of 3 g group (P < 0.01).

CONCLUSIONElimination delay in HDMTX therapy accompanies the suppression of bone marrow and damage of oral mucous membrane, which need more CF rescues and will postpone the following course of chemotherapy. Elimination delay is not associated with the duration of the infusion and the dosage of MTX within the range of 3 approximately 5 g/m2 but there are individual differences.

Adolescent ; Antimetabolites, Antineoplastic ; adverse effects ; pharmacokinetics ; therapeutic use ; Child ; Child, Preschool ; Dose-Response Relationship, Drug ; Female ; Humans ; Infant ; Male ; Metabolic Clearance Rate ; Methotrexate ; adverse effects ; pharmacokinetics ; therapeutic use ; Nausea ; chemically induced ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Retrospective Studies ; Treatment Outcome ; Vomiting ; chemically induced ; Young Adult

OBJECTIVETo determine the distribution characteristics of cancerous foci in the prostate by retrospective analysis on the radical prostatectomy (RP) samples from patients with prostate cancer diagnosed by single positive core biopsy and treated by RP.

METHODSThirty-seven patients with prostate cancer diagnosed by ultrasound-guided biopsy and single positive core biopsy underwent RP. We reviewed the pre- and post-operative data of the patients, compared the results of biopsies and pathological examination of the RP samples, and analyzed the factors that led to the underestimation of the overall prostate cancer risks.

RESULTSPost-operative pathological results showed multifocal distribution of the tumors in 70% of the patients (26/37) and obviously increased Gleason score (7-9) in 56% (21/37). The clinical stages of the tumors had been significantly underestimated preoperatively. The underestimation of their clinical stages might be due to a larger proportion of cancer tissues in a single positive core biopsy, and that of the overall cancer risks attributed to PSAD > 0.2 microg/L. Larger prostate volume (> or = 40 ml) increased the possibility of multifocal distribution.

CONCLUSIONThe risk of prostate cancer diagnosed by single positive core biopsy might be underestimated, and the cancerous foci were characterized by multifocal distribution in the prostate.

Aged ; Biopsy, Needle ; methods ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prostate ; pathology ; Prostatic Neoplasms ; pathology ; Retrospective Studies

OBJECTIVEMonitoring of minimal residual disease (MRD) is proven to be increasingly valuable for predicting relapse and outcome of childhood acute lymphoblastic leukemia (ALL) and is used to identify patients' risk groups in several current clinical trials. However, the limitation is that most studies focused on the cut-off value at 10(-4) and the time point after induction. The aim of this study was to investigate the predictive values of different MRD levels detected at different chemotherapy phases in childhood ALL.

METHODSOne hundred and two patients were enrolled in this study from January 2002 to December 2004 in our hospital. All the patients were treated with modified National Protocol of Childhood ALL in China 1997. MRD levels were detected on the 15th day, 29th day, at 3 months, 6 months and 12 months after initial chemotherapy. All samples were stained with a panel of four colour combinations of fluorochrome conjugated monoclonal antibodies according to the leukemia-associated immunophenotype (LAIP) defined at diagnosis and analyzed by multi-parametric flow cytometry. CD45CD19CD34CD10, CD45CD19CD34CD20 and CD45CD19CD10CD20 were the most common combinations in B lineage ALL, while CD45CD2CD3CD7 and CD45CD2CD3CD34 were the most frequently used immunophenotypes for T lineage ALL. The median follow-up time was 63.3 months ranged from 40.6 to 87.5 months.

RESULTSOf the 102 patients, 64 were male and 38 were female, with a median age of 5.7 (0.2 - 14.8) years. Eighty-eight cases were diagnosed as B lineage ALL and the remaining 14 were T-ALL. The 5-year overall survival (OS) rate and event free survival (EFS) rate for this cohort were (86.9 +/- 3.4)% and (79.9 +/- 4.0)%, respectively. Twelve patients underwent relapse. Among the 102 patients, 14.3% had negative MRD (MRD < 10(-4)) on day 15, 43.9% on day 29, 39.1%, 39.7% and 45.6% had negative MRD at the third, sixth and twelfth month after chemotherapy. Patients who could achieve negative MRD within one year had superior outcome to the others [5-year EFS rates: (92.5 +/- 3.2)% vs. (58.3 +/- 8.6)%, P < 0.001]. The EFS for patients based on MRD levels measured at different stages of therapy were compared by Kaplan-Meier analyses. MRD was predictive of outcome at all 5 time points at a range of thresholds. The optimum threshold, selected for each time point on the basis of log rank analysis, progressively dropped from 10(-2) of day 15 [5-year EFS rates (79.8 +/- 10.3)% vs. (28.6 +/- 17.1)%, P < 0.001], to 10(-3) of day 29 [5-year EFS rates (88.3 +/- 4.9)% vs. (51.3 +/- 14.4)%, P < 0.003], to 10(-4) at 3 [5-year EFS rates (92.4 +/- 5.1)% vs. (65.5 +/- 7.5)%, P < 0.015], 6 [5-year EFS rates (96.3 +/- 3.6)% vs. (65.4 +/- 7.5)%, P < 0.003] and 12 [5-year EFS rates (100.0 +/- 0.0)% vs. (67.7 +/- 8.4)%, P < 0.002] months. And the hazard ratios for relapse and death at higher MRD level groups were 5.91 (95%CI: 1.9 - 18.9), 5.02 (95%CI: 1.5 - 16.5), 5.21 (95%CI: 1.2 - 22.9) and 11.10 (95%CI: 1.5 - 84.5) on day 15, day 29, at month 3 and month 6, respectively. And MRD >or= 10(-2) on day 15 was proven to be an independent predictor by multivariate Cox proportional-hazards regression model.

CONCLUSIONDynamic MRD detection by multi-parametric flow cytometry is highly predictive of outcome for childhood ALL, and the cut-off values at different time points were different.

Adolescent ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Infant ; Infant, Newborn ; Male ; Neoplasm, Residual ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; pathology ; Predictive Value of Tests ; Prognosis

OBJECTIVETo observe the effect of postoperative transcatheter hepatic arterial chemoembolization (TACE) and thymosin alpha(1) (T(alpha1)) treatment on recurrence of hepatocellular carcinoma (HCC).

METHODSFrom Jan 2000 to Dec 2002, 57 patients with HCC were randomly divided into three groups: group A (n = 18) received hepatectomy plus postoperative TACE and T(alpha1), group B (n = 23) received hepatectomy plus postoperative TACE and group C (n = 16) received hepatectomy only. The recurrence rate, the time to tumor recurrence and the median survival for the three groups were investigated.

RESULTSFor group A, B and C, the 1 year recurrence rate was 83.3%, 87.0% and 87.5% (P = 0.926), respectively. The time to tumor recurrence was 7.0, 5.0 and 4.0 months (P = 0.039), respectively. The median survival was 10.0, 7.0 and 8.0 months (P = 0.002), respectively.

CONCLUSIONPostoperative TACE plus Talpha(1) treatment for HCC patients does not decrease the recurrence rate but may delay its occurrence and prolong surviving time.

Adjuvants, Immunologic ; administration & dosage ; Adult ; Aged ; Antibiotics, Antineoplastic ; administration & dosage ; Antineoplastic Agents ; administration & dosage ; Carboplatin ; administration & dosage ; Carcinoma, Hepatocellular ; surgery ; therapy ; Chemoembolization, Therapeutic ; Doxorubicin ; administration & dosage ; Female ; Hepatectomy ; Humans ; Iodized Oil ; administration & dosage ; Liver Neoplasms ; surgery ; therapy ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Neoplasm Recurrence, Local ; prevention & control ; Postoperative Period ; Survival Rate ; Thymosin ; administration & dosage ; analogs & derivatives

Adolescent ; Coronary Angiography ; Coronary Vessel Anomalies ; pathology ; surgery ; Female ; Fistula ; congenital ; pathology ; surgery ; Humans ; Male ; Middle Aged

OBJECTIVELeukemia is the most common hematopoietic malignancies in children. Chemotherapy is currently the primary modality of treatment for this fatal disease. Although chemotherapy is very effective in terms of cell killing, severe side effects such as severe infections, intracranial hemorrhage etc. are frequently encountered due to its poor selective damage between normal and malignant cells or tissues. Thus, a new therapy with highly selective killing of malignant cells which leaves the normal cells unaffected is desperately desired. The aim of this study was to investigate the targeting efficacy in vitro with a new clone of anti-human CD19 antibody immunotoxin 2E8-Genistein on B lineage leukemia cell line Nalm-6 cells and its mechanisms in order to provide the evidence of target therapy on B lineage leukemia and lymphoma.

METHODS2E8-Genistein immunotoxin was generated by conjugating Mab 2E8 with a tyrosine kinase inhibitor, Genistein (Gen) via the Sulfo-SANPAH, an ultra-violet sensitive reagent. Nalm-6, a CD19+ B cell leukemia cell line, was used as target cells, while Molt-3, a CD19-T cell leukemia cell line, was taken as the negative control. The morphology of the cells was observed under the reverted reversed light microscope and the viability was checked with either trypan blue exclusion or MTT methods. Two-color flow cytometry was applied to study the mechanism of cell killing.

RESULTSAfter 24 hours of culture, 2E8-Genistein showed marked target killing on Nalm-6 cells at nine different concentrations from 20 nmol/L through 100 nmol/L with cell survival rates from (71.8 +/- 7.9)% down to (16.6 +/- 12.9)%, respectively (n = 3), which were all significantly lower than that of control group (100 +/- 13.9)% (P < 0.05). The killing effect was even more significant when the concentration was over 80 nmol/L. The growth inhibition rates of this immunotoxin on Nalm-6 cells were 82%, 84% and 94%, respectively at 24, 48 and 72 hours of culture in a time dependent manner. Significant difference was observed between the cell growth curve of Nalm-6 cultured with 100 nmol/L of 2E8-Gen and those of Nalm-6 cultured with medium (blank), PBS (negative control) or the same concentration of pure 2E8 antibody (negative control) groups (F = 152.15, P = 2.15 x 10(-7)), but there was no significant difference among the three control groups (F = 1.51, P = 0.29). When Molt-3 cells were used as target cells, the cell growth curves of Molt-3 cultured with 2E8-Gen (100 nmol/L) and with negative control of blank did not show any significant difference (F = 0.34, P = 0.59). PI/FITC Annexin V double staining analysis with flow cytometry showed that the positive rate (33.45 +/- 8.77)% of early apoptosis on Nalm-6 cells induced by 100 nmol/L of 2E8-Genistein was significantly higher than that of negative control of blank (10.44% +/- 1.28%, t = -4.39, P = 0.001) at 24 hours of culture.

CONCLUSION2E8-Genistein immunotoxin can significantly target the Nalm-6 cells in vitro in a time response manner and the apoptosis induction is involved in the course of this killing effect.

Antibodies, Monoclonal ; immunology ; pharmacology ; Antigens, CD19 ; Apoptosis ; drug effects ; Cell Line, Tumor ; Flow Cytometry ; Genistein ; immunology ; pharmacology ; Humans ; Immunotoxins ; immunology ; pharmacology ; Leukemia, B-Cell ; immunology

OBJECTIVETo observe the effect of postoperative anti-viral therapy using lamivudine and thymosin alpha1 on recurrence of hepatocellular carcinoma (HCC) coexisting with active hepatitis B.

METHODSFrom Jan. 2000 to Dec. 2002, 33 HCC patients with coexisting with active hepatitis B were randomized into two groups: Group I (n = 17) received hepatectomy only, and Group II (n = 16) received hepatectomy and postoperative therapy using lamivudine plus thymosin alpha1. The suppression of HBV-DNA, HBeAg seroconversion rate, tumor recurrence rate and median survival in the two groups were observed and compared.

RESULTSIn Group II and Group I, the 1-year HBV-DNA suppression rate was 100.0% vs 6.0% (P < 0.01), HBeAg seroconversion rate was 62.5% vs 5.9% (P < 0.05), tumor recurrence rate was 81.3% vs 95.5% (P > 0.05), the recurrence time was 7.0 vs 5.0 months (P < 0.01) and median survival 10.0 vs 7.0 months (P < 0.01).

CONCLUSIONAnti-viral therapy using lamivudine and thymosin alpha1 postoperatively may suppress the HBV reaction, delay the recurrence and prolong the survival for patients with HCC with coexisting active hepatitis B.

Adult ; Aged ; Carcinoma, Hepatocellular ; surgery ; therapy ; virology ; DNA, Viral ; drug effects ; Female ; Hepatectomy ; methods ; Hepatitis B ; genetics ; therapy ; Humans ; Lamivudine ; therapeutic use ; Liver Neoplasms ; surgery ; therapy ; virology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Postoperative Period ; Reverse Transcriptase Inhibitors ; therapeutic use ; Survival Rate ; Thymosin ; analogs & derivatives ; therapeutic use

The aim of this study was to evaluate the value of CD117/CD11b phenotypic analysis to diagnosis and prognosis of acute promyelocytic leukemia (APL). Three- or four-color flow cytometry with a series of 22 monoclonal antibodies and CD45/Side Scatter (SSC) gating strategy were used to identify immunophenotypic characteristics of APL as compared to CML in chronic phase (CML-CP). PML/RAR alpha fusion gene was detected by using reverse-transcription polymerase chain reaction (RT-PCR) technique. The results showed that MPO, CD13 and CD33 were almost expressed in all patients with APL and CML-CP whereas HLA-DR and CD34, the hematopoietic progenitor cell markers, were rarely expressed. The positive rate of CD15 in APL was significantly lower than those in CML-CP (P < 0.01). CD117 was positive in 78.3% of the APL cases and in none of the cases of CML-CP. On the other hand, CD11b was almost positive in all cases of CML-CP, but only 16.9% of the APL cases were found positive for this antigen. The CD117+ CD11b- phenotype was present in 72.3% of APL cases while none of cases with CML-CP with this phenotype. Almost all of the cases with CML-CP had the phenotype of CD117- CD11b+. CD117- CD11b+ phenotype was detected in all patients recovering from APL with CD117+ CD11b- phenotype at diagnosis and after treatment with all-trans-retinoic acid (ARTA) for 2 months. PML/RAR alpha fusion gene was positive in 80.6% (25/31) of the APL cases, of which, 64% of the cases belonged to the type L while only 36% of the cases were showed type S for this fusion gene. The positive rates of CD117 were 87.5%, 44.4% and 33.3% in type L group, S group and negative group respectively. It is concluded that analysis of both CD117 and CD11b phenotype may be helpful to the diagnosis, therapy and prognosis of APL in children and adults and to differentiation of APL from recovering benign myeloid proliferation.
Adolescent ; Adult ; CD11b Antigen ; analysis ; Child ; Female ; Humans ; Immunophenotyping ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; diagnosis ; genetics ; immunology ; Leukemia, Promyelocytic, Acute ; diagnosis ; genetics ; immunology ; Male ; Oncogene Proteins, Fusion ; genetics ; Prognosis ; Proto-Oncogene Proteins c-kit ; analysis

OBJECTIVEAcute leukemia (AL) is one of the most common malignant diseases in children. AL immunophenotypes are known to be benefit to the predictable prognoses and specific therapy. Recently, the accuracy of AL immunophenotyping was dramatically improved with the application of the flow cytometry, the new monoclonal antibodies, the improvement of the gating strategies and the multi-parameter analytic techniques. The aim of this study was to evaluate the value of multi-color flow cytometry in the immunophenotyping of acute leukemia in children.

METHODSThree- or four-color flow cytometry and CD(45)/Side Scatter (SSC) gating were used to analyze the surface and cytoplasmic (Cy) antigen expressions in 222 successive cases of childhood acute leukemia.

RESULTSCells from 222 cases of children with acute leukemia were analyzed. Based on the diagnostic criterion proposed by European Group for the Immunological Characterization of Leukemia (EGIL), four categories could be identified: the undifferentiated type accounted for 0.9%, acute myeloid leukemia (AML) 35.1%, acute lymphoblastic leukemia (ALL) 55.9%, and mixed lineage AL 8.1%. Of 124 patients with ALL, 94 patients (75.8%) were classified as B lineage and 30 patients (24.2%) T lineage ALL. Antigen aberrant expressions were found in AML (24.4%), B lineage ALL (36.2%) and T lineage ALL (30.0%). CD(7) was the most commonly expressed lymphoid antigen in AML (12.8%), followed by CD(19) (6.4%) and CD(2) (5.1%). CD(13) was the most commonly expressed myeloid antigen in ALL (18.5%), followed by CD(15) (11.3%), CD(11b) (6.5%) and CD(33) (4.3%). CD(117) and CD(56) presented in 73.3% and 38.0% cases of AML, respectively, but were generally absent on blast cells of ALL. CyCD(22), CyCD(3) and CyMPO were specifically expressed in B lineage, T lineage and myeloid lineage leukemia, respectively, and the first two could be more sensitively detected than they were on cell membrane surface.

CONCLUSIONSMulti-color flow cytometry is a reliable technique in the diagnosis, differential diagnosis and classification of acute leukemia in children.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Infant ; Leukemia ; classification ; diagnosis ; immunology ; Male