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Objective: To investigate the oxidation mechanism of walnut kernels. Methods: Walnut kernels were subjected to vacuum or non-vacuum packaging before accelerated oxidation at 60 ℃. Samples were taken periodically for ultrastructural and morphological observation by transmission electron microscopy (TEM) and scanning electron microscopy (SEM) and for the measurement of oxidation indicators such as peroxide value, carbonyl value, and cell membrane permeability. Changes in cellular ultrastructure during the oxidation process of walnut kernels were analyzed and compared under vacuum and non-vacuum packaging conditions. The oxidation process was assessed from a multi-dimensional perspective and the oxidation mechanism was explored as well. Results: Under non-vacuum conditions, walnut kernel oil began to become sticky on the 14th day of accelerated oxidation and then gradually became stickier. With oxidation time, intracellular protein bodies (PB) exhibited sequential changes including an increase in surface roughness, aggregation, movement toward the cell periphery, decomposition and reassembly. The oil body membrane (OBM) was ruptured and the intracellular reticulum was disrupted, thus causing oil leakage into the intercellular space and resulting in a reduction in the cell volume. Cellular structural changes were much smaller under vacuum conditions than under non-vacuum conditions, the oil started to become sticky on the 28th day, and the oxidation indicators were significantly lower than those under non-vacuum conditions (P < 0.01). The trend of changes in three oxidation indicators was consistent with that of cell ultrastructure. Conclusion: TEM and SEM combined with oxidation indicators can provide clearer insights into cellular ultrastructural changes in walnut kernels after oxidation. The oxidation mechanisms of vacuum and non-vacuum packaged walnut kernels are the same, but their oxidation degrees are different. Notably, vacuum packaging can significantly reduce the oxidation of walnut kernels.

The aim of this study was to develop and investigate the significance of a new multi-factor risk score system to predict the outcome of patients with hematological malignancies received allogeneic hematopoietic stem cell transplantation (allo-HSCT). The impact of pre-, peri-, and post-transplant factors on the outcome including overall survival (OS), disease-free survival (DFS), relapse and transplant-related mortality (TRM) after allo-HSCT were retrospectively analyzed in 122 patients with hematological malignancies at our center. A new risk score system based on the independent risk factors was established and tested. The results showed that absolute monocyte count at day 30 after transplantation (AMC-30, ≥ 536 cells/µl) [hazard ratio (HR) = 0.313, 95% confidential interval (CI):0.156-0.63], WT1( ≥ 1.0%) (HR = 3.268, 95% CI:1.644-6.499), pre-transplant risk grouping (HR = 1.999, 95% CI = 0.993-4.023) were independent prognostic factors of OS and DFS. Patients were divided into 3 groups based on the risk scoring system:group A (no risk factor; score 0), group B (1 risk factor; score 1) and group C (2-3 risk factors; score 2-3). OS at 5 years were 95.1% ± 3.4%, 62.9% ± 6.6% and 36.1% ± 9.6%, respectively (P < 0.0001). DFS at 5 years were 92.6% ± 4.9%, 60.4% ± 6.8% and 15.4% ± 7.1%, respectively (P < 0.0001). The akaike information criterion(AIC) value of the new score system for OS was 331, less than those of AMC-30, WT1, and pre-transplant risk group (346, 343, 346), AIC value for DFS and relapse were 378 and 231, both less than the three single elements(417, 397, 411 and 268, 238, 257). It is concluded that the risk scoring system based on AMC-30, WT1, pre-transplant risk grouping is more highly predictive for clinical outcomes of allo-HSCT than any one of the three single elements.

Objective: To evaluate the efficacy and safety of modified busulfan (Bu, 9.6 mg/kg)/fludarabine (Flu) conditioning regimen on malignant hematologic diseases in elderly and/or drug-intolerable patients., Methods: We utilized a new reduced intensity conditioning (RIC) containing of new dosage of intravenous Bu (9.6 mg/kg), Flu (150 mg/m(2)), cytarabine and semustine but without antithymocyte globulin (ATG) in 23 aged and/or intolerable patients with malignancies. All 23 patients, with a median age of 49 (8-66) years, received an allogeneic hematopoietic stem cell transplantation with human leukocyte antigen (HLA) identical sibling donors during January 2008 and January 2012. Stem cells were collected from granulocyte colony-stimulation factor (G-CSF) mobilized bone marrow plus G-CSF mobilized peripheral blood(G-PB) in 20 patients, G-PB alone in two, and non-mobilized BM in one. The graft had a median mononuclear cells (MNC) of 7.03 (4.04-9.9)×10(8)/kg and 1.76 (0.31-6.43)×10(6)/kg of CD(+)34 cells. Graft-versus-host disease (GVHD) prophylaxis included cyclosporin A, mycophenolate mofetil and methotrexate., Results: All patients were well tolerated to the regimen without serious drug related toxicity. Transplant related mortality were 0 and 4.3% at day 100 and one year. All patients except one got full donor engraftment and achieved complete remission. Acute GVHD was observed in 6 patients (26.1%) including grade III-IV in 5 patients. Chronic GVHD was reported in 15 of 21 evaluable patients (71.4%). With a median follow-up of 1138 (55-1747) days, 16 of 23 patients were alive and disease-free. Three-year overall survival (OS) and event free survival (EFS) were 79.2% and 69.6%, respectively., Conclusion: Modified Bu/Flu as a new RIC regimen is well tolerated and safe for patients who need allogeneic hematopoietic stem cell transplantation, especially in older patients and/or patients with severe comorbidities.

Objective: To analyze the clinical characteristics of the late-onset severe pneumonia after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: A retrospective study was conducted in patients diagnosed as late-onset severe pneumonia after allo-HSCT from March, 2009 to January, 2013 in People's Hospital of Peking University., Results: Of 1538 patients receiving allo-HSCT, 20 developed late-onset severe pneumonia with an incidence rate of 1.3%. Among the 20 patients, 17(85%) had human leukocyte antigen (HLA) identical donors. The other 3(15%) patients had received haplo-identical transplantation. Severe pneumonia occurred at a median time of 227(150-690) days after allo-HSCT. Blood gas tests showed that 14 patients (70%) had arterial oxygen lower than 60 mm Hg (1 mm Hg = 0.133 kPa). All patients had abnormal findings in chest X-ray or CT scan images, 18 (90%) of whom represented bilateral infiltrative lesions. None of the patients had evidence of active graft-versus-host disease before or during pneumonia. Eighteen patients did not respond to empiric antibiotics, neither did 8 patients to targeted medications who had positive pathogens by culture. Eleven patients died of respiratory failure and 9 survived. Six patients received donor lymphocyte infusion (DLI), 4 of whom obtained improvement of chest images and 3 survived till the end of follow-up., Conclusions: The late-onset severe pneumonia after allo-HSCT represents an aggressive behavior with poor prognosis. DLI might be an effective way to improve its outcome.

Objective: To analyze the prevalence of Epstein Barr Virus (EBV) in patients following allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: We retrospectively analyzed the clinical characteristics of 720 patients received allo-HSCT from January 2010 through December 2011 in the Stem Cell Transplant Center of People's Hospital., Results: Of 720 patients (469 male presented and 251 females), with a median age of 30 years (range, 2 to 67 years) old, 66 patients were presented with EBV reactivation. The cumulative incidence of EBV reactivation was (9.3±1.1)%, with a median days of 54.5 (range, 18 to 253 days). During one- year post-transplantation, the cumulative incidences of EBV reactivation in sibling allo-HSCT, haploidentical HSCT and unrelated donor HSCT were (1.3±0.7)%, (13.7±1.7)%, and (9.1±4.4)%, respectively. In patients with haplo-identical HSCT, the cumulative incidences of EBV viremia, probable EBV disease, and post-transplant lymphoproliferative disease (PTLD) were (5.8±1.1)%, (5.7±1.1)%, and (2.3±0.7)%. The mortality was (33.9±5.9)% in all patients with EBV infection: (63.6±15.8)% in PTLD, (42.3±9.9)% in probable EBV disease, (13.8±6.5)% in EBV viremia. By univariate and multivariate analysis, the use of ATG was an independent risk factor for EBV infection., Conclusion: EBV reactivation is a common complication in patients with allo- HSCT, especially high mortality in PTLD and probable EBV disease. The use of ATG was an independent risk factor for EBV infection.

Objective: To investigate the risk factors and prognosis of transplant-associated thrombotic microangiopathy (TA-TMA) following acute graft-versus-host disease (aGVHD), and to evaluate the factors that might influence the prognosis of TA-TMA., Methods: A nested case-control study was designed. Cases with TA-TMA (n = 33) and controls (n = 77) matched for age at allogeneic hematopoietic stem cell transplantation (allo-HSCT) and length of follow-up were identified from a cohort of 356 patients who suffered from aGVHD after allo-HSCT between 2009 and 2011., Results: The median time to presentation of TA-TMA was 3.5 (1.2-23.0) months post-HSCT. The median time from diagnosis and first-line treatment failure of aGVHD to TA-TMA diagnosis was 25 (7-257) days and 15 (5-257) days, respectively. aGVHD occurring beyond 60 days after allo-HSCT, initial grade III-IV aGVHD, first-line treatment failure and receiving tacrolimus as second-line treatment were independently associated with the occurrence of TA-TMA, and patients with two or more risk factors were at higher risk (OR = 210.0, P = 0.000). Twenty-two (66.7%) TA-TMA patients died. Progressive TA-TMA was the significantly adverse factor affecting the survival of TA-TMA cases. None of therapies could improve prognosis of patients with TA-TMA., Conclusion: Many characteristics of aGVHD were associated with TA-TMA, which help us to identify the individuals who are at higher risk of developing TA-TMA following aGVHD and to select the more reasonable GVHD therapeutic strategies.

Objective: To retrospectively review the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for chronic myelomonocytic leukemia (CMML)., Methods: The engraftment, graft versus host disease (GVHD), infection, relapse and survival of 12 CMML patients received allo-HSCT were observed. The clinical outcome of allo-HSCT for CMML was analyzed., Results: Twelve (7 males and 5 females) CMML patients with a median age of 39 years old received allo-HSCT including 7 from HLA-matched sibling and 5 from haploidentical related donors. All 12 patients achieved engraftment. The median time of neutrophil engraftment and platelet engraftment were 15 (11 - 20) days and 13 (11 - 18) days, respectively. 4 patients occurred acute GVHD, and 3 occurred chronic GVHD. After the median follow-up of 17.5 months (12 - 32 months), the overall survival, disease free survival and relapse rate were 66.7%, 66.7%, and 16.7%, respectively., Conclusion: Allo-HSCT can improve the survival of patients with CMML, and is a effective therapy for CMML.

Objective: To investigate the efficacy and safety of rituximab on Epstein-Barr virus (EBV) disease post allogeneic hematopoietic stem-cell transplantation., Methods: A retrospective analysis was performed based on clinical data of 26 patients diagnosed as EBV disease and received rituximab from June 2006 to March 2012 in People's Hospital, Beijing University. Eleven patients were diagnosed as posttransplant lymphoproliferative disorders (PTLD) by histopathology and remaining 15 were diagnosed as probable EBV disease. Patients received a rituximab dose of 375 mg/m(2) once a week. Efficacy was evaluated as revised response criteria for non-hodgkin lymphoma (NHL), and side effects during infusion were evaluated by Common Terminology Criteria for Adverse Events., Results: Patients received 78 infusions with a median of 3(1-6) infusions in each. There were no severe side effects during the infusion of rituximab. The 1(st), 2(nd), 3(rd), 4(th), 8(th) week cumulative complete remission (CR) were (11.5 ± 6.3)%, (42.2 ± 10.2)%, (64.4 ± 10.0)%, (74.6 ± 9.4)%, (87.3 ± 7.9)%, respectively. The overall response rate was 84.5%, and the CR rate was 73.1%. The CR rate was higher among patients with single organ involved than those with multiple organs involved (10/10 vs 9/16, P = 0.023). The CR rate was higher in patients with probable EBV disease than those with PTLD (13/15 vs 6/11, P = 0.095), while there was no statistically significant difference. The incidence of one-year and two-year overall survival since onset of rituximab were (55.7 ± 10.2)% and (39.6 ± 12.4)%, respectively. Survival rate was higher among the patients with single organ involved than those with multiple organ involved (8/10 vs 5/16, P = 0.041). Survival rate was higher in patients with probable EBV disease than those with PTLD (11/15 vs 2/11, P = 0.015)., Conclusions: Rituximab appears to be safe and effective for EBV disease. Due to a potential good response in probable EBV disease, we suggest rituximab should be given based on probable EBV disease; meanwhile the pathological results should get early if possible. Prospective trial is needed to provide evidence so as to define optimal therapy of rituximab.

Objective: To explore the outcome of human leukocyte antigen (HLA)-mismatched/haploidentical hematopoietic stem cell transplantation (HSCT) for refractory/relapsed acute leukemia (AL) patients and its related risk factors., Methods: 96 refractory/relapsed AL patients who received HLA-mismatched/haploidentical HSCT following conditioning regimen comprised of modified busulfan/cyclophosphamide (BU/CY) plus thymoglobulin (ATG) from Jan 2003 to Jun 2011 were analyzed retrospectively., Results: Of the 96 patients, 61 suffered from acute myeloid leukemia (AML), and 35 acute lymphoid leukemia (ALL), all of them in non-remission (NR) or relapse before transplantation. With a median follow-up of 373 (34 - 3157) d, 33 cases (34%) survived, 31 survived without leukemia, and 35 relapsed. The estimated 3-year overall survival (OS) and disease-free survival (DFS) rate was 30.2% and 29.0%, respectively. The 3-year OS rate was significantly higher for AML patients (39.2%) than for ALL patients (15.4%) (P = 0.005). The estimated 3-year OS probabilities for patients with and without prophylactic donor lymphocyte infusion (DLI) were 38.0% and 11.8%, respectively (P = 0.001). Sex, age, conditioning regimen (BU/CY or not, dosage of ATG), the number of HLA mismatches between the donor and recipient, and the number of infused mononuclear cells were not independent factors affecting OS, DFS and relapse. Multivariate analysis showed that DFS rate was significantly higher in patients receiving prophylactic DLI (P = 0.003), in patients with AML (vs with ALL) (P = 0.037) and with chronic GVHD (P = 0.006)., Conclusions: Haploidentical HSCT may prolong DFS in part refractory/relapsed AL patients and even cure them. Prophylactic DLI may reduce relapse and increase survival; for patients with refractory/relapsed ALL, other therapy for prevention and treatment of post-transplant relapse should be explored.

Objective: To investigate the incidence and pathogenesis of fever within the first 24 hours following allogeneic peripheral stem cell transfusion and to analyze the associated risk factors., Methods: Totally 114 patients received allogeneic hematopoietic stem cell transplantation (allo-HSCT) between October 2009 and August 2010 were enrolled and clinical data of febrile patients within 24 hours following peripheral stem cell transfusion were retrospectively analyzed. Multivariate logistic regression analysis was performed to identify the risk factors for transfusion related fever., Results: Thirty-two (28.1%) out of the 114 patients had a fever within 24 hours after allo-HSCT. All of them were human leukocyte antigen (HLA) mismatch transplantation. The median time of the temperature elevated was 2.5 (0 - 18.0) hours after the infusion with a median time of the peak of 7.8 (3.5 - 23.0) hours after the infusion. Fever was attributed to definite infection in 6 patients and no definite infection in the remaining 26 patients. None of them were hemolytic, which was attributed to transfusion related fever. Multivariate analysis showed that female donor and high count of peripheral leukocyte of donor peripheral blood were significant predictors for transfusion related fever., Conclusions: Most of post-infusion fever within 24 hours after HLA mismatch related transplantation has no identifiable infectious focus. The risk factors for transfusion related fever are female donor and high number of peripheral leukocyte of donor blood.

Objective: To analyze the epidemiologic features of blood stream infection (BSI) during the period of agranulocytosis after hematopoietic stem cell transplantation (HSCT) in our hospital, and provide the basis for the empirical therapy., Methods: The consecutive hematopoietic stem cell transplantation recipients conducted between January 2008 and October 2010 were studied retrospectively, to identify the current incidence, etiology for BSI and associated mortality during the period of agranulocytosis., Results: Totally 75 patients were diagnosed as BSI. The incidence of BSI was 9.6% (75/784) in HSCT during the period of agranulocytosis, 17.0% (75/441) in febrile patients. The pathogen testing showed that 64.4% were G(-) bacteria, 30.1% were G(+) bacterial and 5.5% were fungi. All G(-) bacteria except one were sensitive to carbapenems; all G(+) bacteria except one were sensitive to vancomycin. Among the 75 patients, 9 (12.0%) experienced septic shock and 5 (6.7%) died., Conclusions: The pathogens of the BSI in the cohort are G(-) bacteria, followed by G(+) bacteria and fungi. Carbapenems and vancomycin are the first drugs for the experiential therapy.

Objective: To explore the incidence, diagnosis and treatment of nephrotic syndrome (NS) following allogeneic hematopoietic stem cell transplantation (allo-HSCT) in one single center., Methods: Analysis were carried out based on the records of 1464 patients following allo-HSCT hospitalized in Peking University People's Hospital from January 1996 to April 2009, so as to evaluate the incidence and outcomes of NS. A total of 1464 patients whose survival time was longer than 100 d were enrolled. Eight patients (0.6%) had NS following allo-HSCT. Using the nested case-controlled study, 56 cases who received transplantation within the same period were selected as control group for risk factors analysis., Results: The median time of NS was 488 (54 - 1185) d after allo-HSCT. Lower extremities' edema and heavy proteinuria were present in all the patients. Two patients were accompanied with impaired renal function. Six patients suffered from chronic graft-versus-host disease (GVHD). Three patients with NS were accompanied with other symptoms of GVHD. Renal biopsy examination showed typical features of minimal change diseases in 3 patients and membranous nephropathy in 1 patient. All cases achieved complete remission (CR) after taking combined immunosuppressant agents. One patient relapsed soon after all the immunosuppressant agents were stopped. Six patients were still surviving with disease free. One patient died of the relapse of leukemia and another patient died of early discharge from hospital after allo-HSCT. The possible factors correlated with the occurrence of NS were analyzed, and the occurrence of chronic GVHD was identified as an independent factor of NS., Conclusions: NS should be carefully taken into consideration when edema and proteinuria are present in patients following allo-HSCT. The occurrence of NS is probably related with chronic GVHD. NS responds well to immunosuppressant agents.

Objective: To investigate the safety and efficacy of second allogeneic hematopoietic stem cell transplantation for the relapsed hematologic malignancies., Methods: The data of 25 relapsed patients received the second allogeneic transplantation as a salvage therapy in Institute of Hematology Peking University between October 1999 and March 2010 were analyzed retrospectively. Twenty-four patients relapsed at 8.8 (1 - 55) months after the first transplantation, except one received the second transplantation as prophylaxis therapy. They received the second transplantation after 3 (0.3 - 20) months' therapy. The median time between the 2 transplants was 10.6 (0.6 - 59.0) months., Results: Most of the patients were given the conditioning regimen including total body irradiation (TBI, 700 - 779 cGy) or modified busulfan and cyclophosphamide (BUCY, BU 12 mg). All patients survived more than 30 days and achieved sustained white blood cell engraftment. Sinus obstructive syndrome, irradiation dermatitis and acute myocardial infraction were occurred in 3 patients and recoverable. Until January 31 in 2011, with a median observation period of 9.1 (2.0 - 131.1) months, 8 patients had been living with a overall survival (OS) of 30.9%. Twelve patients relapsed at a median 4.4 months and 10 died of it. The other 7 patients died of transplant related complications. The non-relapsed mortality was 35.1%. The disease status at the 2nd transplantation was the only factor which effected the OS (P = 0.009)., Conclusions: The second allogeneic transplantation is a viable option for patients relapsing after the first transplantation. Reduced intensive conditioning regimen ensures the graft engraftment and reduces transplant related toxicity.

Objective: To study the efficacy and safety of human leukocyte antigen (HLA) mismatched hematopoietic stem cell transplantation (HSCT) on severe aplastic anemia (SAA)., Methods: From January 2006 to May 2010, 17 patients received mismatched HSCT. HLA antigens were 3-loci-mismatched in 9 patients, 2-loci-mismatched in 8. All patients received recombinant human granulocyte colony-stimulating factor (rhG-CSF) primed bone marrow cells plus peripheral blood stem cells after modified busulfan/cyclophosphamide + antithymocyte immunoglobulin (BU/CY + ATG) conditioning regimen., Results: All patients achieved full donor type engraftment. Grade III-IV graft versus host disease (GVHD) occurred in 3 patients and extensive chronic GVHD in 1. With a median following-up time of 285 (60 - 1670) d, 11 patients were alive, 9 of them had normal blood counts and the other 2 were blood transfusion independent. Six patients died of transplant-related complications., Conclusion: Mismatched HSCT is a feasible and safe option for SAA patients without sibling identical donors.

Objective: To explore the dissociation of graft-versus-leukemia (GVL) effects from graft-versus-host disease (GVHD) in the patients who experienced GVHD during leukemia relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: The primary disease, disease status, GVHD, response to donor lymphocyte infusion (DLI) and prognosis were analysed in 11 leukemia patients who relapsed with GVHD after allo-HSCT., Results: Of the 11 relapsed, 5 were acute lymphoblastic leukemia and 6 acute myeloid leukemia. Five received DLI before relapse and all developed post-DLI GVHD, including 2 grade II acute GVHD (aGVHD), 1 limited chronic GVHD (cGVHD) plus grade II aGVHD, and 2 extensive cGVHD. After relapse of the 5 patients, 2 received Chemo-DLI, one achieved CR with extensive cGVHD and then relapsed again, the other didn't achieved CR. The other 6 patients didn't received DLI before relapse and also developed post-HSCT GVHD while relapsing, including 3 extensive cGVHD, 1 grade I aGVHD and 2 grade II-IV aGVHD. After relapse, these 6 patients received Chemo-DLI, 2 achieved CR and then relapsed again, 4 didn't achieved CR., Conclusion: The elicited GVHD after allo-HSCT may not accompany effective GVL effects inhibiting leukemic relapse.

Objective: To investigate the characteristics of liver enzyme between acute graft-versus-host disease (aGVHD) and non-aGVHD groups after allogeneic hematopoietic stem cell transplantation (allo-HSCT)., Methods: The liver enzyme data of patients with or without aGVHD was analyzed., Results: Among the 371 patients, 158 developed aGVHD (41.6%) with a median time of 29.5 d. In non-aGVHD group, the median elevating times of ALT, AST, GGT, LDH were all ≤ 20 d after transplantation, which were significantly earlier than those of aGVHD group. The peak value of AST was much higher in aGVHD group, but the remaining liver enzyme showed no significant difference. In aGVHD group, the duration of AST, GGT and LDH was significantly longer than the non-occurrence of aGVHD group, but ALT and ALP showed no difference in duration. In ALT, AST, ALP and LDH elevating group, the occurrence rate of aGVHD was significantly higher, but only ALT and LDH elevation could enter logistic regression model. The sensitivity and veracity of ALT or LDH elevating only were not very good for the diagnosis of aGVHD, but if ALT and LDH both elevated after day 24 and persisted more than 22 d, the sensitivity and veracity were better., Conclusions: The change of liver enzyme is common in allo-HSCT and associates with the occurrence of aGVHD, and the liver enzyme elevating only may not be a good diagnostic index of aGVHD. If the dynamic characteristics of liver enzyme is taken into account and the effect of drug-induced liver injury could be excluded, the elevation of liver enzyme still have the diagnostic value of aGVHD.

Objective: To investigate the correlation between the change of liver enzymes and the severity of acute graft-versus-host disease (aGVHD)., Methods: The liver enzyme and severity of aGVHD in 82 patients were analyzed retrospectively., Results: Among the 82 aGVHD patients, 7 developed grade I (18.3%), 47 grade II (57.3%), 13 grade III (15.9%) and 7 grade IV (8.5%) aGVHD. The elevation of ALT, AST and ALP was found in 49 (59.8%), 36(43.9%) and 8(9.8%)patients respectively. Among these patients, 34 (69.4%) were ALT > 40 U/L, 18 (50%) AST > 40 U/L and 4 (50%) ALP > 132 U/L before the onset of aGVHD. The elevation of ALT and AST was more common and the peak value was significantly higher in patients with grade III-IV aGVHD than with grade I-II aGVHD, but those of ALP showed no difference between the two groups. Compared with the patients without ALT elevation, the proportion of severe aGVHD was higher in patients with ALT elevation., Conclusions: The alteration of liver enzymology is common in aGVHD and its level is correlated with the severity of aGVHD. Liver enzymes, especially the ALT, may be used as a predictor of aGVHD.

Objective: To compare mobilization kinetics of CD34 + cells and MNC during mobilization with either recombinant human granulocyte colony-stimulating factor (rHuG-CSF) filgrastim (Huierxue) or topneuter (Teerjin) in allogeneic hematopoietic stem cells transplantation (allo-HSCT) donors., Methods: Two different agents of rHuG-CSF, Huierxue (filgrastim) and Teerjin (topneuter), were used for hematopoietic stem cells (HSC) mobilization. The HSCs mobilized by each agent were compared in mobilization efficacy, quality of stem cells, side effects in donors and influence of hematopoietic reconstitution in 52 subjects following allo-HSCT. Fifty-two HLA-matched sibling donors, receiving either (rHuG-CSF) topneuter (treatment group) or filgrastim (control group), were randomized in this study., Results: The peak values of both groups, occurring at both Day 4, were (38.9 +/- 3.0) x 10(9)/L and (37.8 +/- 2.9) x 10(9)/L respectively (P > 0.05). Both agents had the following effects upon the HSC parameters: (4.8 +/- 0.7) x 10(8)/kg vs (5.1 +/- 0.4) x 10(8)/kg MNC, (1.6 +/- 0.3) x 10(6)/kg vs (1.9 +/- 0.7) x 10(6)/kg) CD34 + in peripheral blood of donors; (3.8 +/- 0.5)/L vs (3.9 +/- 0.7)/L MNC; (1.3 +/- 0.7) x 10(6)/kg vs (1.5 +/- 0.4) x 10(6)/kg CD34 + in bone marrow of donors (both P > 0.05). With regards to the effects upon hematopoietic reconstitution in subjects following allo-HSCT, both had rapid platelet count recovery (14.6 +/- 0.9) vs (15.2 +/-1.6) days (P > 0.05) and neutrophil count recovery (18.1 +/- 0.8) vs (17.0 +/- 1.9) days (P > 0.05). The adverse events were not significantly different in the HSC donors for these two mobilization agents., Conclusion: Filgrastim and topneuter demonstrate no statistically significant difference in either HSC mobilization kinetics of peripheral and bone marrow blood or hematopoietic reconstitution. Topneuter can be a reliable agent for donor HSC mobilization and recipient hematopoietic reconstitution following allo-HSCT.

Objective: To investigate the relationship between PPAR coactivator 1 (PGC-1), nuclear respiratory factor (NRF), mitochondrial transcription factor A (mtTFA) expressions of vascular smooth muscle cells (VSMC) and development of atherosclerosis in a rabbit model., Methods: Atherosclerotic model was established by feeding the rabbits with high-fat diet for 4, 8 and 12 weeks (n = 10 each). Another 8 rabbits fed with normal diet served as normal controls. Intima-media ratio, mRNA and protein expressions of PGC-1, NRF, mtTFA and SMemb, a marker for synthetic VSMC, were detected on aorta specimens., Results: With the blood lipid increased, the intima-media ratio rose from (0.031 +/- 0.010) microm up to (0.814 +/- 0.258) microm during 12 weeks. Increasing SMemb means that synthetic VSMC grew more and more. The expressions of PGC-1 became significant after 4 weeks (P < 0.01), while that of NRF-1 and mtTFA rose significantly after 8 weeks (P < 0.01)., Conclusions: The PGC-NRF-mtTFA pathway might play a critical role in VSMC proliferation and development of atherosclerosis.

Objective: To study the distribution of Candida spp. in the patients with high-risk of fungal infection and the risk factors of deep candidiasis., Methods: A prospective cohort study was performed among 440 consecutive hospitalized patients admitted to the hematology wards, geriatric wards, and ICUs from May 2004 to April 2005. Stool, urine, and saliva were cultured during the period 72 - 96 h after hospitalization for the first time and then once a week till the patient was discharged or by the end of the sixth week. If deep fungal infection was suspected culture of blood, sputum, bacterium-free body fluid, and/or biopsy specimens were cultured. Medical records were reviewed to analyze the possible risk factors., Results: 426 strains of Candida spp. were isolated from 152 patients, with Candida albicans accounting for 67.4% and other Candida spp for 32.6%. 61 patients were discovered to express Candida colonization. The major species isolated from patients with Candida colonization was Candida albicans. The risk factors identified included two or more broad-spectrum antibiotic administration (odds ratio 16.204; 95% confidence interval, 2.005 to 130.980), Candida colonization (10.636; 3.743 to 30.222), and urinary canal administration (4.285; 1.399 to 13.127)., Conclusion: Candida albicans is still the major organism isolated from the high risk fungal infection patients. Two or more broad-spectrum antibiotic administration, Candida colonization, and urinary canal administration are proved to be the risk factors, with the broad-spectrum antibiotic administration exhibiting more influence than Candida colonization and urinary canal administration.

Objective: To develop a novel platform for detection of the JAK2V617F mutation in patients with myeloproliferative disorders (MPD) by real-time quantitative PCR., Methods: TaqMan-MGB probe was constructed. Peripheral blood samples were collected from 374 MPD patients, 76 with polycythemia vera (PV), 38 with chronic myelogenous leukemia (CML), and 115 with essential thrombocythemia (ET), and 19 with idiopathic myelofibrosis (IMF). Peripheral blood samples from 65 patients with acute myelogenous leukemia (AML), 30 patients with acute lymphoblastic leukemia, 8 patients with chronic lymphoblastic leukemia, and 7 patients with non-Hodgkin's lymphoma and 16 cases of normal donor bone marrow were used as controls. Genomic DNA or RNA was extracted and reversely transcrtibed into cDNA. TaqMan-MGB probe was used to detect the JAK2V617F mutant in MPD. Furthermore, 168 specimens underwent allele specific PCR and 8 specimens underwent sequencing. This method was used on both DNA and cDNA specimens from 38 MPD patients simultaneously so as to test the consistency., Results: The JAK2V617F mutation rates of the PV, ET, and IMF patients were 53 (70%), 59 (51%), (58%) respectively. JAK2V617F mutation was found in only one of the 65 AML patients and was not identified in other control specimens. Both the results of allele specific PCR and of sequencing were consistent with the result of TaqMan-MGB probe method., Conclusion: JAK2V617F mutation is widespread in Chinese MPD patients. Real-time quantitative PCR with TaqMan MGB probe can be used for rapid and accurate detection of the JAK2V617F mutation.

Aim: To study the effects of Fumonisin B1(FB1) on HLA-I on human peripheral blood mononuclear cells (PBMC) in vitro., Methods: The expression of HLA-I on PBMC by FB1 pretreatment at different dosage(10, 50 micromol/L) was detected with flow cytometry (FCM), Western blot, and semi-RT-PCR, respectively., Results: The expression of HLA-I on PBMC in vitro at the two experimental concentration was both lower than that of the control after FB1 treatment for 24 h as represented by fluorescence intensity by FCM analysis. Western blot results further confirmed the above results. At mRNA level, HLA-A, B and C mRNA were detected by RT-PCR, and the results showed that no changes were found on the expression of HLA-A, B mRNA between FB1 treated group and the control group, but HLA-C mRNA was inhibited in FB1 treated groups., Conclusion: 10 and 50 micromol/L FB1 could inhibit the expression of HLA-I on human PBMC in vitro at 24 h treatment.

Objective: To study the pharmacokinetics of multi-dose oral tetra-arsenic tetra-sulfide (As(4)S(4)) in acute promyelocytic leukemia (APL) patients and its major side effects., Methods: Clinical pharmacokinetic study of As(4)S(4) was carried out in 7 patients who had never used arsenic before. The total concentration of arsenic in blood, urine and hair was determined with hydride generation-atomic absorption spectrometry., Results: These patients were administrated oral As(4)S(4) complex capsule 20 mg/kg three times a day for 14 days. From day 10 and to day 14, the blood arsenic concentration and urinary arsenic excretion reached a steady state. The average minimal concentration (Cmin) and maximal concentration (Cmax) of blood arsenic was (53.3 +/- 9.0) microg/L and (70.7 +/- 10.8) microg/L. t 1/2 was prolonged to (70.7 +/- 31.7) hour, and area under curve (AUC) was (448.9 +/- 71.8) microg. h(-1).L(-1). Median time to peak concentration (Tmax) was 1 (range 0.5 - 8) hour. During As(4)S(4) therapy, 24-hour arsenic content in urine was (6170.8 +/- 3141.8) microg/L and it accounted for about (0.152 +/- 0.082)% of the total daily dosage. It decreased steadily over time after drug withdrawal, arsenic accumulated in hair and the concentration could be ten-fold higher than that before treatment., Conclusion: Multi-dose oral As(4)S(4) is safe and has been relatively well tolerated in APL patients in spite of the tendency of its retention in some tissues after long time administration.

Objective: To analyze on the efficacy and toxicity of fludarabine and teniposide + mitoxantrone (MIT) regimens on treating refractory and relapsed acute lymphocytic leukemia in adult patients., Methods: Teniposide 100 mg/d, 5-7 d, MIT 10 mg/d, 2 d and fludarabine regimens [Flu 30 mg/(m(2) . d), 3- 5 d, Cytarabine (Ara-c )1-2 g/(m(2) . d), 5 d; Flu 50 mg/d, 5 d, Ara-c 200 mg/d, 5 d, MIT 4 mg/d, 4 d] were used to treat 42 cases of adults with refractory and relapsed acute lymphocytic leukemia(ALL). G-CSF 5 microg/(kg . d) were used when WBC<1.0 x 10(9)/L., Results: In both the regimens fludarabine and VM (teniposide + MIT), the complete remission (CR) rate was 45% versus 31.8% (P>0.05); the median neutropenia began 6 days after the regimens arresting and lasting 10 versus 7.5 days, P>0.05; thrombocytopenia begin at time of 10 versus 6.5 days (P<0.05) after the regimens arresting and lasting 6 versus 10 days (P>0.05). Fludarabine regimen had less non-haematological toxic effect than that of VM., Conclusion: Compared with VM, Fludarabine regimen was a very effective alternative treatment for CR induction in adult patients with refractory and relapsed ALL and low toxicity.

Objective: To explore the prognostic significance of expression of survivin and caspase-3 in esophageal squamous-cell carcinoma (ESCC) and the relasionship with expression of heat shock proteins 27 and 70 (HSP27 and HSP70)., Methods: Expressions of survivin and caspase-3 in 101 cases of ESCC were quantitatively detected with flow cytometry. Their expressions in long-term survival group (group A, >or= 5 years, 38 cases) were compared with those in the short-term survival group (group B, 0.05). The positive expression rate of survivin in group A was significantly lower than that in group B (31.6% vs 54.0%, P = 0.029). Compared with that in short-term survival group, the strong positive expression rate of caspase-3 in long-term survival group was significantly higher (47.6% vs. 68.4%, P = 0.042). Positive expression rate of caspase-3 showed decreasing tendency with increase in age. No significant differences in clinicopathologic features in relation to expression rate of caspase-3 other than tumor length. No correlation was observed between expression intensity of survivin and any clinicopathologic features. Logistic regression analysis indicated that survivin and caspase-3 expressions were of independent prognostic significance for ESCC. There was no association between survivin and caspase-3 expression and expression of HSP27 and HSP70., Conclusion: The expressions of survivin and caspase-3 are two independent prognostic factors in ESCC. They do not correlate with HSP27 and HSP70 expression.

Aim: To explore the effects of deoxynivalenol (DON) on transporter associated with antigen processing-1 (TAP-1) expression in human peripheral blood mononuclear cells (PBMCs)., Methods: Effects of various concentrations of DON on TAP-1 expression of in vitro cultured human PBMCs and the dose-effect relationship were analyzed by flow cytometry (FCM) and semi-quantitative RT-PCR at the protein and mRNA levels., Results: FCM analysis indicated that treatment with various concentrations of DON could inhibit TAP-1 expressions in PBMCs, showing a negative correlation between DON concentration and TAP-1 expression. Semi-quantitative RT-PCR detection indicated that high concentrations of DON (1,000 and 2,000 microg/L) could distinctly inhibit TAP-1 mRNA expression., Conclusion: DON can down-regulate TAP-1 expression in human PBMCs in a dose-dependent manner in vitro, which suggests DON may interfere with host immunosurveillance, and this may account for the correlation between DON-contaminated grain and esophagus cancer in high risk area.

Background & Objective: Natural killer (NK) cell and dendritic cell (DC) play important roles in anti-tumor immunity. Heat shock protein (HSP) is also involved in anti-tumor immune mechanisms. This study was to explore prognostic significances of NK cells and DCs infiltrations in esophageal squamous cell carcinoma (ESCC), and their relationships with expressions of HSP27 and HSP70., Methods: NK cells, and DCs infiltrations were detected by SP immunohistochemistry in 101 specimens of ESCC. Among the relevant 101 patients, 38 were classified into long-term survival group (>/=5 years), and 63 into short-term survival group ( 0.05, t test]. NK cells and DCs infiltrations in ESCC had no significant correlation with clinicopathologic features, such as length of tumor, depth of invasion, histological grade, and lymph node metastasis, etc. (P > 0.05). Binary logistic regression analysis indicated that infiltration of NK cells was of prognostic significance for ESCC (P=0.001), while DCs was not (P=0.842). Infiltration of NK cells was positively correlated with infiltration of DCs (r=0.266, P=0.007). Both infiltrations of NK cells and DCs were positively correlated with expression of HSP27 (P=0.017, P=0.018), while no such correlation with expression of HSP70 was found (P > 0.05)., Conclusions: Infiltration of NK cells is an independent prognostic factor of ESCC, and might be a prognostic biomarker of ESCC. Infiltration of DCs has no correlation with prognosis of ESCC. Infiltrations of NK cells and DCs may have internal correlation with the expression of HSP27.

Objective: To further explore the carcinogenic activity of Sterigmatocystin (ST) and the possible synergistic carcinogenic effect of deoxynivalenol (DON) in NIH mice., Methods: NIH mice were randomly divided into 6 groups, 30 in each. Five groups of mice were given by gastric intubation ST 3 microg/kg, ST 30 microg/kg, ST 3 microg/kg + DON 1.5 microg/kg, ST 30 microg/kg + DON 1.5 microg/kg and DON 1.5 microg/kg respectively, 3 times a week for 24 weeks. The remaining group of mice was given normal saline accordingly, serving as control. All mice were fed with HPLC-confirmed mycotoxin-free food, analysis. The mice were killed and pathologically examined at 58th and 74th weeks., Results: No pathological changes were found in the control group of mice. Adenocarcinoma of lung was observed in 25.0%, 41.7%, 62.5%, 69.2% and 37.5% of mice given ST 3 microg/kg, ST 30 microg/kg, ST 3 microg/kg + DON 1.5 microg/kg, ST 30 microg/kg + DON 1.5 microg/kg and DON 1.5 microg/kg, respectively. In addition, dysplasia of glandular stomach was detected in 50.0%, 58.3%, 37.5%, 53.8% and 25.0% of mice similarly treated., Conclusion: Oral administration of ST or DON can induce adenocarcinoma in lung and dysplasia of glandular stomach in NIH mice. There is synergistic carcinogenic effect when both ST and DON are given.

Objective: Aflatoxin G1 (AFG1) is a member of the carcinogenic aflatoxin family produced by aspergillus flavus. It is a major contaminating mycotoxin in food in areas of China with high cancer incidence. The purpose of this study is to explore the carcinogenic effects of AFG1 in NIH mice., Methods: NIH mice were randomly divided into three groups. Two experimental groups were treated intragastrically by gavage with AFG1 3 microg/kg and AFG1 30 microg/kg respectively, 3 times a week for 24 weeks. The control group was treated with normal saline. All mice were fed with food that was free of AFGs as confirmed by HPLC analysis. The mice were weighed every week throughout the entire experiment, and then sacrificed and examined pathologically at the 58th and 74th weeks respectively., Results: Compared with control mice receiving no AFG1, bronchial epithelial hyperplasia, alveolar hyperplasia and adenocarcinoma of lung were observed in mice receiving AFG1 treatment. The incidences of bronchial epithelial hyperplasia, alveolar hyperplasia and adenocarcinoma of lung were 60.0%, 10.0% and 30.0% for mice receiving 3 microg/kg AFG1 and 28.6%, 35.7%, 42.9% for mice receiving 30 microg/kg of the toxin, respectively., Conclusion: Oral administration of AFG1 can induce hyperplastic lesions and adenocarcinoma of lung in NIH mice.

In order to explored the change of PML/PML-RARalpha protein during tetraarsenic tetrasulfide (As4S4) treatment, acute promyelocytic leukemia (APL) cells from a group of newly diagnosed APL patients were examined by indirect immunofluorescence staining with anit-PML monoclonal antibody. The results showed that all samples typically presented many microspeckle signals throughout the nucleus before treatment. The redistribution occurred as early as on the second day after As4S4 treatment, which revealed loss of microspeckles with the presentation of a few large speckles. Anti-PML staining also emerged in the perinuclear cytoplasm. At last, microspeckles and large speckles all disappeared. When the therapy was combining all-trans-retinoic acid (ATRA) with As4S4, similar results were obtained. However, APL cells from patients treated with ATRA alone performed totally different appearance, presenting microspeckles and large speckles at the same time, followed with entirely large speckles. The conclusion is that As4S4 makes redistribution of PML/PML-RARalpha protein in leukemic cells from APL patients during the treatment, which is quite different from that during the treatment of ATRA.

Objective: To study the expression of EP-CAM, beta-catenin in the carcinogenesis of squamous cell carcinoma of uterine cervix., Methods: The expressions of EP-CAM and beta-catenin were detected with immunohistochemical stain in 14 cases of normal cervical squamous epithelium, 32 cases of cervical intraepithelial neoplasia (CIN) and 38 cases of cervical invasive squamous cell carcinoma., Results: The over-expression rates of EP-CAM were 0, 7.1%, 20.0%, 62.5% and 55.3% for normal cervical epithelium, CINI, CINII, CINIII and carcinoma groups. The EP-CAM over-expression rates in CINIII and cervical carcinoma groups were significantly higher than those in normal epithelium and CINI groups (P < 0.001). No aberrant expression of beta-catenin was shown in normal cervical epithelium, while the aberrant expression rates of beta-catenin in CINI, CINII, CINIII and cervical carcinoma group were 28.6%, 40.0%, 62.5% and 84.2%. The aberrant expression rate of beta-catenin increased with the increase in degree of CIN and development of cervical carcinoma. The over-expression rate of EP-CAM was reversely related to the differentiation of cervical squamous cell carcinoma (P < 0.001)., Conclusion: EP-CAM and beta-catenin may be involved in the carcinogenesis of squamous cell carcinoma of uterine cervix. The over-expression of EP-CAM and aberrant expression of beta-catenin may serve as markers of squamous carcinogenesis of uterine cervix.

Background & Objective: Cixian County is one of the high incidence areas of esophageal carcinoma in China. Up to now, no work has been conducted on the possible etiological role of human papilloma virus (HPV) infection on esophageal carcinoma in this local area. The aim of this study was to explore the putative roles of HPV infection on the esophageal carcinogenesis of the patients in Cixian and to elucidate the possible relationship between HPV existence and fragile histidine triad gene (FHIT gene, a putative tumor suppressor gene), expression in squamous cell carcinoma of esophagus., Methods: The existence of HPV DNA and the expression of FHIT gene at protein level in esophageal carcinoma tissues were determined with PCR and immunohistochemical staining, respectively, in 128 archival paraffin-embedded tissue blocks of esophageal squamous carcinoma from the high incidence area of Cixian and 24 tissue blocks from the non-high incidence area., Results: PCR results showed that the positive detection rate of HPV in esophageal carcinoma tissues from the high incidence area was 20.3%, which was slightly higher than that from the non-high incidence area (8.3%)(P >0.05). Immunohistochemically, aberrant expression of FHIT gene in esophageal carcinoma tissue was found in 75.6% cases from the high incidence area and only 54.2% from the non-high incidence area, which was significantly lower than that of the former (P< 0.05). No correlation was found between the abnormal expression of FHIT and existence of HPV DNA in esophageal squamous carcinoma tissues., Conclusion: HPV DNA could be detected in partial esophageal carcinoma cases from the high incidence area of esophageal carcinoma in Cixian. The aberrant expression rate of FHIT protein in the cases from the high incidence area of esophageal carcinoma in Cixian is higher than that from the non-high incidence area.