Rhaponticum uniflorum inhibits H2O2-induced apoptosis of liver cells via JNK and NF-κB pathways / 中国中药杂志

To study the inhibitory effect of Rhaponticum uniflorum on apoptosis induced by H2O2 in HepG2 cells. Human HepG2 cells injury models were established by H2O2, then cell survival rate was assayed by MTT method; levels of LDH, ALT, and AST were detected by chemical colorimetric method;SOD activity was detected by xanthine oxidase method; GSH content was detected by dithio-bis-nitrobenzoic acid(DTNB); MDA level was detected by thiobarbituric acid (TBA) method;and the relative activities of Caspase-3, 8 and 9 were measured by Colorimetry. The expression levels of Cleaved Caspase-3(Casp-3), cytochrome(Cyto c), NF-κB, ERK, JNK, p38 MAPK, as well as the phospharylated proteins were determined with Western blotting method. The results showed that R. unifloru had no significant effect on cell viabilities of HepG2 cells at the concentrations of 25-400 mg•L⁻¹. However, H2O2decreased the cell viabilities, increased the cellular oxidative stress, and up-regulated the protein expressions of Casp-3, cytoplasmic Cyto c, p-JNK and nuclear NF-κB. As compared with the model group,R. unifloru could increase the cell viability, reduce LDH, ALT and AST leakage, reduce the MDA formation, increase the SOD and GSH levels,reduce the relative activities of Caspase-3, 8 and 9, down-regulated the protein expressions of Casp-3 and cytoplasmic Cyto c, and down-regulate the p-JNK and nuclear NF-κB levels.The results indicated that R. unifloru had the inhibitory effect on apoptosis induced by H2O2in HepG2 cells, and the mechanism maybe associated with inhibiting JNK activation and NF-κB nuclear translocation.

Protective effect of soyasaponins on acute liver injury induced by D-galactosamine and lipopolysaccharide in mice / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of soyasaponins on acute liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in mice.</p><p><b>METHOD</b>The mice were randomly divided into five groups: the normal control, the model group, the silymarin (positive control) group, and soyasaponins high and low-dose groups. They were administered with drugs once every day for 7 days. At the end of the experiment, GalN and LPS were injected intraperitoneally to all of the groups except for the normal group to establish the acute liver injury model. The pathological changes were detected with hematoxylin & eosin (HE) staining, tumor necrosis factor-alpha (TNF-alpha) was detected by ELISA method, and the alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), reduced glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), and the activation of Caspase-3 and Caspase-8 were detected by the colorimetric method.</p><p><b>RESULT</b>Soyasaponins could reduce the activities of serum ALT and AST, the acute hepatic injury induced by GalN/LPS, serum TNF-alpha level, hepatic NO and MDA contents, and the Caspase-3 and Caspase-8 activations of liver tissues, and increase the hepatic CAT, GPx, GST and GSH levels.</p><p><b>CONCLUSION</b>Soyasaponins shows the protective effect on acute liver injury induced by GalN and LPS in mice, which may be related to its antioxidative ability and anti-liver apoptosis.</p>

Intervention effect of aqueous fractions from Boschniakia rossica on hepatic oxidative stress in mice with liver injury induced by carbon tetrachloride / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the intervention effect of aqueous fractions from Boschniakia rossica (BRAF) on hepatic oxidative stress in mice with liver injury induced by carbon tetrachloride (CCl4).</p><p><b>METHOD</b>The experimental mice were randomly assigned into the normal control group, the model group, the silymarin (positive control) group, as well as high and low dose BRAF groups. Mice were treated intragastrically with silymarin or BRAF once every day for 7 days. At the end of the experiment, CCl4 was injected intraperitoneally into the mice to establish the acute liver injury model. The pathological changes was detected with hematoxylin and eosin (HE) staining, and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), superoxide dismutase (SOD) , catalase (CAT), glutathione peroxidase (GPx), Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, and the contents of reduced glutathione (GSH) and malondialdehyde (MDA) were detected by the colorimetric method.</p><p><b>RESULT</b>BRAF significantly reduced ALT, AST and ALP activities in serum, alleviated hepatic injury induced by CCl4, increased SOD, CAT, GPx and GSH levels in liver, and SOD, Na + -K + -ATPase and Ca2+ -Mg2 + -ATPase activities in liver mitochondria, and decreased the MDA content in liver and liver mitochondria.</p><p><b>CONCLUSION</b>BRAF reduces hepatic oxidative stress in mice with acute liver injury induced by CCl4, thereby showing the protective effect on mice with acute liver injury induced by CCl4.</p>

[Flow cytometric monitoring of minimal residual diseases in patients with acute leukemia after allogeneic hemapoietic stem cell transplantation].

OBJECTIVE: To study the significance of flow cytometric monitoring minimal residual diseases (MRD) in patients with acute leukemia (AL) after allogeneic hemapoietic stem cell transplantation (HSCT). METHODS: From January 2007 and January 2008 MRD were detected by flow cytometry (FCM) in 402 bone marrow (BM) in 102 AL patients without leukemic gene and chromosomal changes at first diagnosis after HSCT (1, 2, 3, 6,12 months after HSCT; adding detection frequency in part of high risk patients), The relationship between the MRD results and clinical prognosis were observed. Patients with significantly higher MRD were treated and the effectiveness was monitored by FCM (MRD > 0.01% considered as positive). RESULTS: (1) 71 cases were persistently negative for MRD after HSCT and all them were in hematologic complete remission (CR). Only 3 cases had extramedullary relapse. The disease free survival (DFS) and overall survival (OS) were 66.2% and 90.1%, respectively. (2) Of 27 MRD(+) cases 11 converted to MRD negativity after chemotherapy plus donor lymphocyte infusion (DLI), CIK, NK cells. The DFS and OS were 63.6% and 72.7%, respectively. Other 16 cases had hematologic relapse. The DFS and OS were 11.1% and 25.0%, respectively. The median time from MRD increasing to hematologic relapse was 48 days (7-69 day). (3) Four cases had hematologic relapse after HSCT and died in the end. CONCLUSIONS: (1) The DFS and the OS in MRD(-) cases are significantly higher than those of MRD(+) cases. (2)MRD(+) patients after HSCT coveted to MRD(-) after intervention. Therapy, whose DFS and the OS are still significantly higher than those of MRD(+) cases. (3) Patients with hematologic relapse after HSCT have the worst prognosis and the DFS and OS are significantly low. FCM monitoring of MRD in patients after HSCT is a sensitive, specific, quick and simple method. It can indicate recurrent state in time, facilitates early intervention, reduces the hematologic relapse risk and improves DFS.

[Veto T cells enhance engraftment allogeneic hematopoietic stem cells].

Veto activity was defined as the capacity of specifically downregulating cytotoxic T-precursor cell (CTL-p) directed against antigens of the veto cells themselves but not against third-party antigens. Many studies have shown that the most potent veto cells are CD8(+) cytotoxic T lymphocytes (CD8(+)CTLs). Effectively, CD8(+)CTLs of donor origin can facilitate engraftment of donor's stem cells by eliminating host-alloreactive T lymphocytes. In this article, effect mechanisms, depletion of GVH ex vivo, application in vivo, synergistic enhancement with rapamycin and regulatory T cells, and anti-tumor effect in the hematopoietic stem cell transplantation are summarized.

[Monitoring of early Epstein-Barr virus reactivation and preemptive therapy after allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To investigate early Epstein-Barr virus (EBV) reactivation and the outcome of preemptive therapy after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: From January 2007 to January 2009, totally 277 patients after allo-HSCT were studied (haploidentical 116, unrelated 75, matched sibling 86). Conditioning regimens were mainly busulfan (BU) + cyclophosphamide (CY)/fludarabine (Flu) or total body irradiation (TBI) + CY/Flu. Antihuman thymocyte globulin (ATG) was added in haploidentical and unrelated transplants. Plasma EBV DNA was monitored once to twice weekly in the first 3 months after allo-HSCT with real time quantitative polymerase chain reaction (RQ-PCR). EBV viremia was diagnosed when EBV DNA was more than 5 × 10(2) copies/ml but without symptoms. Acyclovir (10 mg/kg, intravenous drip, 8 h) was used for preemptive therapy and immuno-suppressants were decreased if possible. RESULTS: Totally 33 patients (11.9%) developed EBV viremia with a median time at day 44 (day 19 to day 84). The incidences of EBV viremia in the transplants from matched sibling, haploidentical, unrelated donors were 0, 15.5%, 20.0%, respectively. There was no significant difference between haploidentical and unrelated transplants (P = 0.09), but much less EBV viremia was seen in matched sibling transplant (P = 0.001). Twenty of 33 patients (60.6%) had complete response to preemptive therapy. The median time to reach EBV DNA negative in plasma was 11 (4 - 56) d. The median duration of preemptive therapy was 21 (14 - 60) d. Both univariate and multivariate analysis indicated that haploidentical and unrelated transplants, acute graft versus host disease (GVHD) were the risk factors for EBV viremia. Two-year overall survival in the patients with EBV viremia was significantly lower than that without EBV viremia (54.2% vs 72.1%, P = 0.006). CONCLUSIONS: Our large clinical study has demonstrated that preemptive therapy with acyclovir that is guided by EBV viremia is effective in majority of the patients with high-risk for EBV reactivation after allo-HSCT, which may further decrease the risk for developing life-threatening EBV disease or post-transplantation lymphoproliferative disorder. Haploidentical and unrelated transplants, acute GVHD are the risk factors for EBV viremia which has negative impact on survival.

[Cytomegalovirus enteritis after allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To analyse the clinical features, diagnostic methods and risk factors of cytomegalovirus (CMV) enteritis after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: Analysis was made on 24 cases of CMV enteritis after allo-HSCT in Beijing Daopei Hospital from Aug. 2007 to Jul. 2009, including clinical data, endoscopic diagnosis, histopathological and virological results, and the association between CMV enteritis with viremia and graft-versus-host disease(GVHD). RESULTS: 87.5% of the patients were over 18 years old. The median time to diagnosis of CMV enteritis was 63 days after HSCT. The mucosal lesions in enteroscopic examination had no significant differences between CMV enteritis and gastrointestinal GVHD complicated with the enteritis. The methods used in diagnosis included histopathology (32.1%) and virology (92.9%). The copies of CMVDNA in mucosal samples greater than 10(5)/10(6) PBNC was better diagnosis. A number of risk factors were compared between the survival and death groups: type of transplant, conditioning regimen, the time span of ganciclovir prophylaxis therapy, grade II-IV GVHD before enteritis, the time of diagnosis as GVHD, using MP > or = 1 mg/kg to treat GVHD, the time between GVHD and enteritis, CMV viremia before enteritis, the time of diagnosis as enteritis, CMVDNA quantitation, and there were no any statistic differences. CONCLUSION: Cytomegalovirus enteritis should be carefully diagnosed by histopathology and virology through endoscopic examination. It is better to undertake pan-colon endoscopy as well as terminal ileum examination for more accurate diagnosis. PCR can significantly improve the detection rate. CMVDNA detection in patients' stool may be helpful to diagnosis, especially for those patients who can not stand the endoscopy examination.

[Clinical research on reconstitution of CD4+CD25+ T cells after haploidentical bone marrow transplantation].

The aim of this study was to investigate the reconstitution of CD4(+)CD25(+) T cells after haplo-identical bone marrow transplantation (hiBMT) and its correlation with graft versus host disease (GVHD) and relapse. Peripheral blood samples from 27 patients after hiBMT were harvested and the percentage and absolute counts of CD4(+)CD25(+) T cells were detected by flow cytometry. The correlations of GVHD occurrence and disease relapse with the reconstitution of CD4(+)CD25(+) T cells were analyzed. The results showed that the percentage of CD4(+)CD25(+) T cells of peripheral blood samples increased significantly after G-CSF priming. At day 30 after hiBMT, CD4(+)CD25(+) T cells were recovered to the 20% of normal level, followed by a slowly process in 3 months, and up to one half of the normal level at 180 days. There was no evidence to prove relationship between CD4(+)CD25(+) T cells and acute GVHD, while CD4(+)CD25(+) T cells were increased significantly in the chronic GVHD group. The absolute count of CD4(+)CD25(+) T cells showed no relations with relapse of leukemia during the first year after hiBMT. In conclusions, chronic but not acute GVHD was in relation to the reconstitution of CD4(+)CD25(+) T cells based on the anti-CD25 antibody therapy model for the prevention of GVHD after hiBMT. Further investigation is needed to clarify whether the relapse of leukemia after hiBMT is related to the reconstitution of CD4(+)CD25(+) T cells.

[Correlation between CD34+CD61+ megakaryocyte precursors and platelet engraftment in allogeneic hematopoietic stem cell transplantation].

This study was purposed to investigate the correlation between the dose infused megakaryocytic precursors (CD34+, CD34+CD61+) and recovery time of platelet count following an allogeneic PBSCT and/or BMT through quantitative detection of CD34+ and its subpopulation in peripheral blood and BM mobilized by G-CSF. 24 patients with various hematologic malignancies received PBSCT/BMT from their HLA matched or unrelated donors and haploidentical siblings in April-December 2007. 20 evaluated patients were divided into 2 groups according to different transplant schemes. HLA matched group received PBSCT regime and haploidentical group received PBSCT combined with BMT. CD34+CD61+ subpopulations in sample from patients receiving PBSCT/BMT were measured by flow cytometry immediately or storage over night. The results showed that the median number of infused CD34+, CD34+CD61+ and CD34-CD61+ cells in haploidentical group were 6.24x10(6)/kg (1.53-20.48), 66.19x10(4)/kg (8.16-493.83), and 34.38x10(6)/kg (14.71-109.16) respectively, in HLA matched group those were 4.88x10(6)/kg (1.00-8.24), 14.16x10(4)/kg (11.63-96.87), and 13.50x10(6)/kg (1.74-35.61), respectively. Median days of ANCs>0.5x10(9)/L and platelets>20x10(9)/L were 18.5 (11.0-29.0) days and 16.5 (9.0-35.0) days in haploidentical group respectively; in HLA matched group those were 14.5 (9.0-24.0) and 10.5 (6.0-37.0) respectively. A significance difference of median days for ANC engraftment presented between two groups (p=0.048). There was no significant difference of time for platelet engraftment between 2 groups. For patients with CD34+ cell dose>2x10(6)/kg there was significant difference of time of platelet engraftment between HLA matched and haploidentical groups (p=0.006). The number of CD34+CD61+ cells infused in 12 haploidentical patients or in 8 HLA matched patients were much better correlated with the time of platelet recovery up to 20x10(9)/L than that of number of CD34+ cells infused in total 20 patients (r=-0.768 and p=0.004 for haploidentical CD34+CD61+ cells, r=-0.747 and p=0.033 for HLA matched CD34+ CD61+ cells, r=-0.449 and p=0.047 for CD34+ cells). There was an inverse correlation between the number of infused CD34+ CD61+ cells and time of platelet engraftment. Therefore, as the number of CD34+ CD61+ cells increased, duration of platelet engraftment (time to reach platelet count of 20x10(9)/L) shortened significantly. It is concluded that the determining the number of megakaryocytic precursor by flow cytometry may predict the platelet reconstitutive capacity of the allogeneic hematopoietic stem cell transplantation, which is in haploidentical PBSCT and in BMT.

[Application of fibrotic bronchoscopy in the diagnosis of pulmonary diffuse infiltration following bone marrow transplantation].

In order to evaluate the diagnostic value of fibrotic bronchoscopy (FB) in the pulmonary infiltration following bone marrow transplantation (BMT), 18 patients with pulmonary complications after BMT from November 2003 to March 2006 were performed with FB. Bronchoalveolar lavage (BAL) and brushing were performed in patients who had received short-term empirical therapy without good response, and transbronchial lung biopsy (TBLB) was carried out in 3 cases. The results showed that 9 out of 10 cases with pulmonary infection, including bacterial pneumonia (n = 3), aspergillosis (n = 2), pneumocystis carinii pneumonia (n = 3) and viral infection (n = 1) were diagnosed by using FB. One case was diagnosed as tuberculosis after open lung biopsy following negative results from twice BAL. 2 out of 8 cases were diagnosed by TBLB as noninfectious pulmonary complications. In conclusion, FB, especially with BAL, is a safe and useful procedure for the evaluation of pulmonary complications, which is particularly suitable for diagnosis of pulmonary infection after BMT. Furthermore, TBLB should be recommended in order to avoid open lung biopsy, if the patients tolerate the operation.

[Clinical investigation on treatment of children leukemia with non-T-cell depleted haploidentical bone marrow transplantation].

To investigate the efficacy and feasibility of parent non-T cell depleted haploidentical bone marrow transplants (haploidentical BMT) for children with leukemia, the efficacy of haploidentical BMT was evaluated in 8 leukemia children (1.9-9 years) received hematopoietic stem cell transplantation, donors were their parents with HLA-mismatched for two or three loci. Five children were pre-conditioned with a myeloablative regimen consisting of high-dose cytarabine (Ara-C), cyclophosphamide (CY) and total body irradiation. Busulfan (BU), Ara-C and CY were used for preconditioning regimen in other three children. The donors were given G-CSF prior to marrow harvest and the non-T-cell depleted grafts were used. A combination of CsA, MTX, ATG, mycophenolate mofetil (MMF) and CD25 monoclonal antibody were used for GVHD prophylaxis. The results showed that rapid engraftment was observed in all cases after transplantation by cytogenetic evidence. The mean time of neutrophil count exceeded 0.5 x 10(9)/L and the mean time of platelet count exceeded 20 x 10(9)/L were 16 and 17 days after transplantation respectively. Incidence of lethal aGVHD was lower, II-III acute aGVHD was found only in one out of eight patients. Chronic GVHD was observed in five patients, 4 from which showed local cGVHD, one developed extensive cGVHD. During the follow-up of 33 months (range 7-56 months), two patients died from relapsed leukemia, including one relapsed as donor-origin leukemia. Disease-free survival was achieved in the remaining six patients. No death occurred during the follow-up of 6 months. It is concluded that above-mentioned preconditioning regimen and GVHD prophylaxis procedure in non-T-cell depleted bone marrow transplantation from HLA-mismatched parents are effective approaches and safe strategy for the treatment of children leukemia.

Impact of incompatible killer cell immunoglobulin-like receptor and its ligand on the outcome of haploidentical bone marrow transplantation.

The purpose of study was to investigate the impact of killer immunoglobulin-like receptor (KIR) and its ligand on haploidentical bone marrow transplantation. 74 cases were analyzed for the distribution frequencies and characteristics of KIR and its ligand as well as the impact of KIR ligand for the haploidentical bone marrow transplantation in terms of the overall survival, disease-free survival (DFS), GVHD and relapse. The results showed that among the 19 KIR genotypes currently nominated KIR2DL1, KIR2DL4 and KIR3DL2-3 could be detected in all the cases. Other high frequency genotypes included KIR3DP1 (98.6%), KIR2DP1 (98.6%), KIR3DL1 (97.3%) and KIR2DL3 (97.3%). Inhibitory receptor genotypes were 1.37-fold of activating receptor genotypes. KIR2DL1, KIR3DL2, KIR3DL3 and KIR2DL4 were found in all haplotypes and at least one genotype of KIR2DL2 and/or KIR2DL3 existed in all haplotypes. Among the 14 genotypes found in the test, the HLA-Cw7 was the most popular (37.8%) and the group 2 (HLA-Cw1, 3, 7, 8, 13, 14) recognized by KIR2DL2/2DL3 counted for 43.2%. The incompatibility of KIR for 32 cases of haploidentical BMT was 43.8%, of which 9/14 were KIR2DL incompatible, 5/14 were KIR2DL2 or KIR3DL1 incompatible. Among the 46 cases of haploidentical BMT, 29 cases were HLA-Cw matched and 14 cases were mismatched. The completed mismatch ratio of HLA-Cw was 30.4% and the match ratio was 63.4%. The survival rate was higher for the 14 cases of KIR genotype compatible group than the 13 cases of KIR genotype incompatible group (p = 0.032). The disease-free survival was significantly higher for the 17 cases of mismatched KIR ligands (HLA-Cw) group than the matched group (p = 0.024). The survival rate was higher in GVHD group than that in non-GVHD group when the KIR ligand was missing. The acute and severe GVHD was related to the existence of activating receptor of KIR2DS1/2DS2. The incompatibility group was accompanied with frequent acute and severe GVHD and less relapse and vice versa for the compatibility group. One patient died after BMT among the 14 mismatched KIR ligand group suffering from myelogenous leukemia while 4 patients out of 12 patients died in the matched group. It is concluded that the haploidentical BMT is characterized by mismatch between donor and recipient and its immunological reactions also features by the incompatibility of KIR genotype and missing ligand. The missing ligand for the donor KIR has strong effect on the outcome of BMT and it means a lot to analyze the KIR genotype and its ligand for the selection of best donor and prognostic evaluation in haploidentical BMT.

[Antithymocyte globulin used for treatment of severe acute graft versus host disease after haploidentical bone marrow transplantation].

The objective of study was to investigate the effect of low-dose antithymocyte globulin (ATG) on steroid-resistant severe acute graft versus host disease (aGVHD). Six patients with steroid-resistant severe aGVHD after haploidentical bone marrow transplantation (BMT) received the treatment with ATG at a low dose of 1.25 mg/kg for 3 - 5 doses every other day. The results showed that 3 out of 6 patients got completely remission (CR), among them 2 patients have still been in disease-free survival, 1 patient died from leukemia relapse. 1 out of the other 3 patients got partial remissin (PR), 2 patients were aggravated. The other 3 patients all died from GVHD. The major complications observed in these patients were infections. In conclusion, low-dose ATG is effective for some patients with steroid-resistant severe aGVHD, and has not severe side effect. To strengthen environmental protection should be considered as important for prevention of infection.

[Neurological complications following haploidentical bone marrow transplantation].

UNLABELLED: To explore the occurrence patterns of neurological complications following haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, choices of therapies and prognosis in 10 patients with neurological disorders were summarized. The results showed that complications occurred in 10 out of 74 patients after bone marrow transplantation, which include 3 with encephalorrhagia, 3 infection, 1 epilepsy, 1 Guillian-Barr's syndrome, 1 encephalopathy and 1 schizophrenia. 4 patients died of neurological complications. IN CONCLUSION: neurological complications commonly occurred in haploidentical bone marrow transplantation, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nerve system infection and peripheral nerve diseases associated with slow immune reconstitution should have clinical interests.

[Pulmonary complications in haploidentical bone marrow transplantation].

UNLABELLED: To explore the occurrence patterns of pulmonary complications at different stages in haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, management choices and prognosis in 18 patients with pulmonary disorders were summarized. The results showed that in 18 out of 70 patients after bone marrow transplantation occurred pulmonary complications which included pneumonia affected by bacteria (7 cases), fungus (5 cases) and cytomegalovirus (CMV, 4 cases), bronchiolitis obliterans organizing pneumonia (BOOP, 1 case), and idiopathic pneumonia syndrome (1 case), out of which 8 cases died. Fungal and CMV pneumonia occurred predominantly 2 to 3 months after transplantation, whereas bacterial pneumonia was observed in the duration of 3 to 12 months and 4 cases of them suffered from secondary fungal infections during treatment. BOOP and idiopathic pneumonia syndrome were diagnosed 12 months and 50 days after transplantation respectively. IN CONCLUSION: pulmonary complications were commonly seen in haploidentcal bone marrow transplantation, and fungal pneumonia might be the main cause that needs intensive management.

[CD25 monoclonal antibody for GVHD prophylaxis in non-T-cell depleted haploidentical bone marrow transplantation for treatment of childhood leukemia].

OBJECTIVE: Allogeneic bone marrow transplantation (Allo-BMT) has improved long-term survival in children patients with refractory leukemia. For patients who do not have an HLA-identical sibling, the treatment option is limited. Using related mismatch donors or parental donors for Allo-BMT was at high risk for acute severe GVHD. The purpose of this study was to explore the effects of CD25 monoclonal antibody on reducing the incidence of severe acute GVHD in haploidentical bone marrow transplantation for the treatment of childhood leukemia. METHODS: Ten children with leukemia received haplotype Allo-BMT from HLA two or three loci mismatched related donors. Most patients were classified as in high risk category. The donors of patients were given G-CSF (Lenograstim Chugai) 250 microg/day for seven doses prior to marrow harvest. The non-T-cell depleted haploidentical bone marrow was infused. In addition to combination of CSA, MTX, ATG (Fresenius Hemocare, Germany) and mycophenolate mofetil (MMF) for GVHD prophylaxis, CD(25) monoclonal antibody (Simulect, Novartis Pharma Switzerland) was administered to prevent acute severe GVHD.A total of 40 mg Simulect was given in two doses of 20 mg each by 30 min intravenous infusion 2 h before transplantation and day 4 after transplantation. The outcomes were compared with those of 8 children patients with leukemia who received haploidentical bone marrow transplantation without CD(25) antibody for GVHD prophylaxis. RESULTS: All patients were engrafted and sustained full donor-type engraftment. 100% donors hematopoietic cells after transplantation was determined by cytogenetic evidence analysis. The median days of granulocyte exceeding 0.5 x 10(9)/L and pallets exceeding 20 x 10(9)/L were 19 and 22 days, respectively. Patients were monitored till up to days 100 for the sign of aGVHD. None developed the grade II-IV acute GVHD. However, the incidence of the grade II-IV acute GVHD in control group was 50% (P = 0.0147). Eight patients could be evaluated for chronic GVHD. All experienced chronic GVHD confined to the skin. The median follow-up duration was 12 months (range 9 - 24 months). Two patients died from transplant related mortality, one had patient relapsed disease and died. The remaining seven patients were alive in disease-free situation. CONCLUSION: The use of Simulect in haploidentical bone marrow transplantation for the treatment of children patients with leukemia is effective for preventing acute severe GVHD and may reduce transplant-related mortality.

Anti-atherosclerotic effect of soybean isofalvones and soyasaponins in diabetic rats / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the influence of soybean phytochemical extract containing isoflavones and soyasaponins (SPE) on blood glucose, blood lipids, plasma lipid peroxide and platelet aggregation activity in diabetic rats.</p><p><b>METHODS</b>Diabetic rats were fed with fodder containing 20 g/kg of SPE for 20 weeks. Their plasma very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) were separated by sequential ultracentrifugation.</p><p><b>RESULTS</b>Twenty weeks after experiment, level of blood glucose, atherosclerotic index and plasma level of lipid peroxide were (11.9 +/- 0.9) mmol/L, 0.40 +/- 0.14 and (15.7 +/- 0.5) mmol/L, respectively in diabetic rats fed with SPE, significantly lower than those in control rats not fed with it, (14.2 +/- 2.0) mmol/L, 0.58 +/- 0.22 and (20.7 +/- 3.0) mmol/L, respectively. Accordingly, platelet aggregation rates induced by ADP and collagen in the two groups were (54.1 +/- 8.8)% vs (66.6 +/- 12.4)% and (58.0 +/- 7.9)% vs (69.6 +/- 9.4)%, respectively. Changes in all these indices were significantly different between the two groups.</p><p><b>CONCLUSION</b>SPE could significantly decrease blood glucose, improve atherosclerotic index, and inhibit lipid peroxidation and platelet aggregation in diabetic rats, which might be useful in prevention and control of diabetes mellitus and diabetes-associated atherosclerosis.</p>

[Clinical study of a new protocol for acute graft-versus host disease prophylaxis in HLA-haploidentical bone marrow transplantation (BMT)].

OBJECTIVE: To explore the feasibility of a new protocol for acute graft versus host disease (aGVHD) prophylaxis in haploidentical bone marrow transplantation. METHODS: Thirty-eight high-risk leukemia patients underwent haploidentical G-CSF primed bone marrow transplantation without ex-vivo T cell depletion, the donors were given G-CSF 250 microg/d for 7 days prior to marrow harvest. All patients received a same chemo-radiation conditioning regimen, including cytarabin, cyclophosphamide and total body irradiation (TBI). GVHD prophylaxis regimen consisted of ATG, CsA, MTX, Mycophenolate mofetil (MMF) and anti-CD(25) monoclonal antibody (MoAb). RESULTS: All patients achieved engraftment of a median of 19 and 22 days for neutrophil and platelet, respectively. Cytogenetic analysis showed 100% donor hematopoietic cells in all recipients after transplantation. One of the twenty patients (5%) experienced grades II - IV acute GVHD. The recovery of CD(3)(+)CD(8)(+) T cells, CD(19)(+) cells and CD(56)(+) cells after transplantation was within 3 approximately 12 months. Of the 20 patients, 16 were alive with minimal and limited chronic GVHD and karnofsky score over 90% in a median follow-up of 9 months. Disease free survival (DFS) rates was (80 +/- 9)%. CONCLUSION: G-CSF priming marrow graft along with sequential immunosuppressants, especially the addition of anti-CD(25) MoAb for aGVHD prophylaxis could achieve excellent engraftment, proper immune reconstitution and very low incidence of grade II - IV GVHD. The new protocol is effective and feasible in preventing severe acute GVHD and improving DFS.

Humanized anti-CD25 monoclonal antibody for prophylaxis of graft-vs-host disease (GVHD) in haploidentical bone marrow transplantation without ex vivo T-cell depletion.

OBJECTIVE: To investigate the effects of a novel anti-IL-2 receptor (CD25) monoclonal antibody, basiliximab, on graft-vs-host disease (GVHD) and engraftment in haploidentical bone marrow transplantation (BMT). MATERIALS AND METHODS: Thirteen consecutive high-risk leukemia patients (age 9-41) underwent haploidentical BMT with G-CSF-primed marrow as stem cells without ex vivo T-cell depletion. Basiliximab, along with a combination of cyclosporine (CSA), methotrexate (MTX), and mycophenolate mofetil (MMF), was used for GVHD prophylaxis. Immunophenotyping, limited-dilution assay, and colony-forming assays were used to measure the effect of basiliximab on the subsets of lymphocytes, cytotoxic T-lymphocyte precursors (CTLp), and hematopoietic cells. RESULTS: All patients established successful trilineage engraftment with full donor chimerism. No patients developed grade II-IV acute GVHD. Patients who survived more than 12 months and were free of relapse showed limited chronic skin GVHD. Ten of 13 patients are currently alive with a Karnofsky performance score of 100% at median follow-up of 17 months (range 12-24 months). Basiliximab significantly decreased alloreactive CTLp by 10-fold to 100-fold in limiting-dilution assays. It had no effect on hematopoietic stem and progenitor cells as determined by in vitro colony-forming assays. CONCLUSION: The addition of basiliximab to CSA, MMF, and MTX as GVHD prophylaxis effectively reduced severe lethal GVHD in haploidentical BMT. It is possible to selectively eliminate or reduce the number of alloreactive T cells with anti-CD25 antibody, which results in prevention of or a reduction in the severity of GVHD.