Panaxdiol Saponins Component Promotes Hematopoiesis and Modulates T Lymphocyte Dysregulation in Aplastic Anemia Model Mice.

OBJECTIVE: To investigate the potential efficacy of panaxadiol saponins component (PDS-C) in the treatment of aplastic anemia (AA) model mice. METHODS: Totally 70 mice were divided into 7 groups as follows: normal, model, low-, medium-, high-dose PDS-C (20, 40, 80 mg/kg, namely L-, M-, H-PDS-C), cyclosporine (40 mg/kg), and andriol (25 mg/kg) groups, respectively. An immune-mediated AA mouse model was established in BALB/c mice by exposing to 5.0 Gy total body irradiation at 1.0 Gy/min, and injecting with lymphocytes from DBA mice. On day 4 after establishment of AA model, all drugs were intragastrically administered daily for 15 days, respectively, while the mice in the normal and model groups were administered with saline solution. After treatment, the peripheral blood counts, bone marrow pathological examination, colony forming assay of bone marrow culture, T lymphocyte subpopulation analysis, as well as T-bet, GATA-3 and FoxP3 proteins were detected by flow cytometry and Western blot. RESULTS: The peripheral blood of white blood cell (WBC), platelet, neutrophil counts and hemoglobin (Hb) concentration were significantly decreased in the model group compared with the normal group (all P<0.01). In response to 3 dose PDS-C treatment, the WBC, platelet, neutrophil counts were significantly increased at a dose-dependent manner compared with the model group (all P<0.01). The myelosuppression status of AA was significantly reduced in M-, H-PDS-C groups, and hematopoietic cell quantity of bone marrow was more abundant than the model group. The colony numbers of myeloid, erythroid and megakaryocytic progenitor cells in the model group were less than those of the normal mice in bone marrow culture, while, PDS-C therapy enhanced proliferation of hematopoietic progenitor cells by significantly increasing colony numbers (all P<0.01). Furthermore, PDS-C therapy increased peripheral blood CD3+ and CD3+CD4+ cells and reduced CD3+CD8+ cells (P<0.05 or P<0.01). Meanwhile, PDS-C treatment at medium- and high doses groups also increased CD4+CD25+FoxP3+ cells, downregulated T-bet protein expression, and upregulated GATA-3 and FoxP3 protein expressions in spleen cells (P<0.05). CONCLUSION: PDS-C possesses dual activities, promoting proliferation hematopoietic progenitor cells and modulating T lymphocyte immune functions in the treatment of AA model mice.

Eltrombopag interference in routine chemistry testing.

BACKGROUND: High doses of Eltrombopag have been previously reported to cause bilirubin interference. Following receipt of a sample from a patient receiving high-dose Eltrombopag therapy, the laboratory decided to investigate the effect of this drug on routine chemistry testing. METHODS: Interference studies were performed by spiking Eltrombopag into aliquots of a serum pool to give concentrations ranging from 0 to 500 µg/mL. The following analytes, namely albumin, alkaline phosphatase, alanine transaminase, aspartate transaminase, Urea, total calcium, cholesterol, triglycerides, glucose, high-density lipoprotein cholesterol, iron, magnesium, inorganic phosphate, creatinine, bicarbonate, transferrin, ferritin, electrolytes, total and direct bilirubin and serum indices (hemolysis, icterus and lipaemia) were then measured on the Roche Cobas 6000 chemistry analyzer (Roche, Indianapolis, USA). RESULTS: Eltrombopag interference (>10% change of the baseline value) was observed for total cholesterol, triglycerides, inorganic phosphate and high-density lipoprotein cholesterol. Clinical significant interference was observed for total cholesterol, inorganic phosphate and high-density lipoprotein cholesterol CONCLUSIONS: Presence of high Eltrombopag concentrations in blood samples has been demonstrated to cause interference in the measurement of certain spectrophotometric-based assays on the Roche Cobas 6000 analyzer.

Clinical study of pai-neng-da capsule in the treatment of chronic aplastic anemia.

OBJECTIVE: To evaluate the efficacy and safety of Pai-Neng-Da Capsule (panaxadiol saponins component, PND), a new Chinese patent medicine, on patients with chronic aplastic anemia (CAA) and to explore the optimal therapeutic regimen for CAA. METHOD: A total of 36 patients with CAA were enrolled and divided into three groups: the AP group (20 cases, andriol 120 mg/day + PND 240 mg/day), the ACP group (13 cases, andriol 120 mg/day + cyclosporine 3-6 mg kd(-1) day(-1) + PND 240 mg/day), and the PND group (3 cases, PND 240 mg/day). All patients were treated and followed up for 6 months. Peripheral blood counts, renal and hepatic function and Chinese medical (CM) symptoms of patients were assessed and all indices were gathered at the beginning and end of the study. RESULT: In the AP group, no significant hematologic difference was observed at the end of 6-month treatment comparing with the beginning. In the ACP group, the blood counts were maintained at the same level after the 6-month treatment. In the PND group, trilineage hematologic improvement was displayed at the end of 6-month treatment comparing with the beginning. No significant difference was showed in renal and hepatic function in all patients. All patients' clinical symptom improved according to CM symptom score. The effective rates were 95%, 73% and 100%, respectively. CONCLUSION: PND improved the efficacy and decreased side effects by cutting down the dosage of andriol, and it could also improve patients' clinical symptom and quality of life. PND were effective and safe in the treatment of CAA, it could be used alone or in combination with pharmacological agents such as andriol and cyclosporine.

Recent advances in treatment of aplastic anemia.

Recent advances in the treatment of aplastic anemia (AA) made most of patients to expect to achieve a long-term survival. Allogeneic stem cell transplantation (SCT) from HLA-matched sibling donor (MSD-SCT) is a preferred first-line treatment option for younger patients with severe or very severe AA, whereas immunosuppressive treatment (IST) is an alternative option for others. Horse anti-thymocyte globuline (ATG) with cyclosporin A (CsA) had been a standard IST regimen with acceptable response rate. Recently, horse ATG had been not available and replaced with rabbit ATG in most countries. Subsequently, recent comparative studies showed that the outcomes of patients who received rabbit ATG/CsA were similar or inferior compared to those who received horse ATG/CsA. Therefore, further studies to improve the outcomes of IST, including additional eltrombopag, are necessary. On the other hand, the upper age limit of patients who are able to receive MSD-SCT as first-line treatment is a current issue because of favorable outcomes of MSD-SCT of older patients using fludarabine-based conditioning. In addition, further studies to improve the outcomes of patients who receive allogeneic SCT from alternative donors are needed. In this review, current issues and the newly emerging trends that may improve their outcomes in near futures will be discussed focusing the management of patients with AA.

Comparative metabolomics analysis on hematopoietic functions of herb pair Gui-Xiong by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry and pattern recognition approach.

The compatibility of Angelicae Sinensis Radix (Danggui, DG) and Chuanxiong Rhizoma (Chuanxiong, CX), a famous herb pair Gui-Xiong (GX), can produce synergistic and complementary hematopoiesis. In present study, global metabolic profiling with ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) combined with pattern recognition method was performed to discover the underlying hematopoietic regulation mechanisms of DG, CX and GX on hemolytic and aplastic anemia rats (HAA) induced by acetyl phenylhydrazine (APH) and cyclophosphamide (CP). Thirteen endogenous metabolites contributing to the separation of model group and control group were tentatively identified. The levels of LPCs including lysoPC (18:0), lysoPC (20:4), lysoPC (16:0) and lysoPC (18:2), sphinganine, nicotinic acid, thiamine pyrophosphate, phytosphingosine, and glycerophosphocholine increased significantly (p<0.05) in HAA, while the levels of oleic acid, 8,11,14-eicosatrienoic acid, ceramides (d18:1/14:0), and 17a-hydroxypregnenolone decreased significantly (p<0.05) in comparison with control rats. Those endogenous metabolites were chiefly involved in thiamine metabolism and sphingolipid metabolism. The metabolic deviations could be regulated closer to normal level after DG, CX and GX intervention. In term of hematopoietic function, GX was the most effective as shown by the relative distance in PLS-DA score plots and relative intensity of metabolomic strategy, reflecting the synergic action between DG and CX. The relative distance calculation was firstly used in metabolomics for semi-quantization.

Recent advances in treatment of aplastic anemia

Recent advances in the treatment of aplastic anemia (AA) made most of patients to expect to achieve a long-term survival. Allogeneic stem cell transplantation (SCT) from HLA-matched sibling donor (MSD-SCT) is a preferred first-line treatment option for younger patients with severe or very severe AA, whereas immunosuppressive treatment (IST) is an alternative option for others. Horse anti-thymocyte globuline (ATG) with cyclosporin A (CsA) had been a standard IST regimen with acceptable response rate. Recently, horse ATG had been not available and replaced with rabbit ATG in most countries. Subsequently, recent comparative studies showed that the outcomes of patients who received rabbit ATG/CsA were similar or inferior compared to those who received horse ATG/CsA. Therefore, further studies to improve the outcomes of IST, including additional eltrombopag, are necessary. On the other hand, the upper age limit of patients who are able to receive MSD-SCT as first-line treatment is a current issue because of favorable outcomes of MSD-SCT of older patients using fludarabine-based conditioning. In addition, further studies to improve the outcomes of patients who receive allogeneic SCT from alternative donors are needed. In this review, current issues and the newly emerging trends that may improve their outcomes in near futures will be discussed focusing the management of patients with AA.

Hematologic responses in patients with aplastic anemia treated with deferasirox: a post hoc analysis from the EPIC study.

Reports are emerging of hematologic responses associated with iron chelation therapy; however, studies are limited in aplastic anemia patients. Deferasirox reduced iron overload in aplastic anemia patients enrolled in the EPIC (Evaluation of Patients' Iron Chelation with Exjade(®)) study (n=116). A post hoc analysis of hematologic responses was conducted on 72 patients with evaluable hematologic parameters (according to UK guideline criteria), 24 of whom received deferasirox without concomitant immunosuppressive treatment. Partial hematologic responses were observed in 11 of 24 (45.8%) patients; all became transfusion-independent. One patient had an additional platelet response and one patient had an additional platelet and hemoglobin response. Mean serum ferritin levels at end of study were significantly reduced in partial hematologic responders (n=11; -3948 ± 4998 ng/mL; baseline 6693 ± 7014 ng/mL; percentage change from baseline -45.7%; P=0.0029). In non-responders, the reduction in serum ferritin was less pronounced (n=13; -2021 ± 3242 ng/mL; baseline 4365 ± 3063 ng/mL; % change from baseline -27.6%; P=0.0171). Alongside reduction in iron overload, deferasirox may, therefore, improve hematologic parameters in a subset of aplastic anemia patients. Further investigation is required to elucidate the mechanisms involved.

Effects of sodium copper chlorophyllin on mesenchymal stem cell function in aplastic anemia mice.

OBJECTIVE: To investigate the effects of sodium copper chlorophyllin (SCC) on the proliferation, differentiation and immunomodulatory function of mesenchymal stem cells (MSCs) from mice with aplastic anemia. METHODS: A mouse model of aplastic anemia was established by exposure of BALB/c mice to sublethal doses of 5.0 Gy Co60 γ radiation, followed by transplantation of 2×10(6) lymph node cells from DBA/2 donor mice within 4 h after radiation. Aplastic anemic BALB/c mice were randomly divided into six groups: the treated groups, which received 25, 50, or 100 mg/kg/day SCC, respectively; a positive control group treated with cyclosporine A (CsA); and an untreated model control group (model group); while, the non-irradiated mice as the normal control group. SCC or CsA were administered by gastrogavage for 20 days, starting on day 4 after irradiation. Peripheral blood cells were counted and colony-forming fibroblasts (CFU-F) in the bone marrow were assayed. The ability of MSCs to form calcium nodes after culture in osteoinductive medium was also observed. The immunosuppressive effect of MSCs on T lymphocytes was analyzed by enzyme-linked immunosorbent assay and flow cytometry, to evaluate the efficacy of SCC in mice with aplastic anemia. RESULTS: Peripheral blood white cell and platelet counts were increased by medium and high SCC doses, compared with the untreated control. CFU-Fs were also increased compared with the untreated control, and the numbers of calcium nodes in MSCs in osteoinductive medium were elevated in response to SCC treatment. The percentage of Forkhead box protein 3 (FOXP3(+)) T cells was increased in T cell-MSC cocultures, and the cytokine transforming growth factor ß1 was up-regulated in SCC-treated groups. CONCLUSION: The results of this study suggest that SCC not only promotes the proliferation and differentiation of MSCs, but also improves their immunoregulatory capacity in mice with aplastic anemia.

Restoration of hematopoiesis after iron chelation therapy with deferasirox in 2 children with severe aplastic anemia.

Iron overload is a significant clinical problem in patients with severe aplastic anemia or other transfusion-dependent bone marrow failure diseases. Iron chelation therapy is more readily available owing to the recent introduction of oral iron chelators. We describe 2 cases of children with severe aplastic anemia and related transfusional iron overload who received iron chelation therapy with oral deferasirox. Our patients experienced restoration of trilineage hematopoiesis after the administration of deferasirox along with the reduction in ferritin levels, and subsequently became transfusion-free. Our report raises the possibility of potential benefit on hematopoiesis from iron chelation therapy and warrants furthermore investigations.

[Regulation of erythropoiesis in patients with suppressed hematopoiesis during mountain climatic treatment].

AIM: to estimate the regulation of erythropoiesis and the coagulation system in patients with suppressed hematopoiesis in a mountain hospital (3200 m above sea level). SUBJECTS AND METHODS: The investigation included 12 patients with aplastic anemia (AA) and 10 with idiopathic thrombocytopenic purpura (ITP). Blood was received at a Bishkek hospital, then on days 20 and 40 of stay in the mountains. The authors studied erythropoietin (EPO) by enzyme immunoassay (Protein Contour kit, Russia), serum ferritin (SF) by immunoradioassay (Immunotech kit, Czech Republic), hypoxia-inducible factor-1alpha (HIF-1alpha), homocysteine (HC), hepcidin, endothelin (ET), and thrombomodulin (TM) by sandwich enzyme immunoassay, by applying monospecific antisera and monoclonal antibodies against relevant antigens (IDG Int Inc, USA). RESULTS: On staying in the mountains, there was a gradual increase in the content of hemoglobin in patients with AA and ITP. On day 40, in keeping with higher hemoglobin (Hb) levels, both groups showed a decrease in HIF-1alpha concentrations to the normal values (from 8.2 to 4.5 pg/ml). Due to the anemic syndrome, baseline EPO was increased by 5-7 times in the patients from both groups. On days 20-40, the content of EPO showed a 1.3-2.5-fold increase. In AA, HC was almost 3 times greater than the normal values; in ITP, it was 1.5-fold increased. On day 20 and during the patients'stay in the mountains, the level of HC remained in the normal range in both groups. CONCLUSION: Hypoxic hypoxia positively affects a number of hematological parameters, by normalizing erythropoiesis (Hb, EPO, and HIF-1alpha), iron metabolism (SF), and the coagulation system (HC, ET, and TM).

Iron chelation therapy associated with improvement of hematopoiesis in transfusion-dependent patients.

BACKGROUND: It is well known that iron overload may cause multiple organ failure. In chronically transfused patients, optimal iron chelation therapy is associated with reduced morbidity and mortality. Furthermore, chelation therapy has been associated with erythroid responses. STUDY DESIGN AND METHODS: Among chronically transfused adults affected by myeloproliferative neoplasms and treated with iron chelators, two case reports are described. CASE REPORT: A male adult patient with myelodysplastic syndrome (MDS) and a female adult with aplastic anemia (AA), both transfusion-dependent, were treated with deferasirox, an oral iron chelator. RESULTS: A significant reduction in transfusion requirement was observed and was dependent on chelation therapy. The patient affected by AA also experienced a significant increase in hemoglobin levels. Minimal doses of deferasirox maintained the erythroid responses. Many mechanisms of action of the drug on erythropoiesis have been postulated. The early erythroid response seems to be independent of the removal of iron from deposits, per se, since the reduction of ferritin levels (a surrogate marker of iron deposits) below threshold levels occurs as a later event. CONCLUSION: Although there are few reports on erythroid responses in patients undergoing iron chelation therapy, they may give new insights in the pathogenesis of MDS and other myeloproliferative neoplasms. AA may benefit in terms of erythroid response. The findings in these cases underline the clinical importance of treating patients with iron overload. A survival benefit of chelation in patients with myeloproliferative neoplasms is still to be confirmed.

Hematopoietic activity of Smilax aristolochiaefolia (zarzaparrilla) in mice with aplastic anemia.

Smilax aristolochiaefolia (Liliaceae) has been used in Mexican traditional medicine for the treatment of tumors, leprosy, anemia and as a tonic for skin infections and anemia. Aplastic anemia (AA) was induced in CD1 mice 8-12 weeks old distributed 10 animals each in Groups VSC, AA, AASa and AAr. Groups AA, AASa and AAr received benzene (2 ml/kg diluted v/v with corn oil) subcutaneously every three days until 20 dosages had been administered. The vehicular solution control group (VSC) received corn oil and the HC group (healthy control) received saline solution. Two days after the last benzene inoculation, groups AA and HC were bled and sacrificed to count blood and bone marrow cells. Group AASa received an aqueous S. aristolochiaefolia (0.4 g/kg) solution orally on days 3, 5 and 7 after the last dosage of benzene, meanwhile group AAr received no treatment after induction of AA (self recovery). On day 9 these groups were bled and sacrificed to count blood and bone marrow cells. Mice with aplastic anemia treated with S. aristolochiaefolia extract, recovered normal platelet levels and nucleated bone marrow cells as compared with the control, but the counts of erythrocytes and leukocyte were lower than controls (p<0.005). The aqueous extract of S. aristolochiaefolia (zarzaparrilla) restores hematopoeisis in the bone marrow of mice with aplastic anemia.

[Effect of busui shengxue granule on serum tyrosine kinase 3 ligand and fibronectin in patients with chronic aplastic anemia].

OBJECTIVE: To observe the clinical effect of Busui Shengxue Granule (BSG) in treating chronic aplastic anemia (CAA) and the changes of serum fibronectin (Fn) level and expression of FMS-like tyrosine kinase 3 ligand (FL). METHODS: Sixty-eight patients with CAA were assigned to two groups. The 35 patients in the test group were treated with BSG and the 33 in the control group by Zaizhang Shengxue Tablet. Levels of Fn and FL were determined before and after 6-month treatment by enzyme linked immunosorbent assay. RESULTS: The clinical effective rate in the test group was 74.3% (26/35 cases), and that in the control group 48.5% (16/33 cases), the difference between the two groups was significant (P <0.05). In the test group, patients of yang-deficiency type showed an effect superior to that of yin-deficiency type, 93.8% (15/16 cases) vs 57.9% (11/19 cases, P < 0.05). High expression manners of Fn and FL were shown in all CAA patients, the high expressions of both were lowered to some extent after treatment, but the improvement in the test group was better than that in the control group (P <0.05), and the deviation was more liable to be corrected in patients of Shen-yang deficiency type than that in patients of Shen-yin deficiency type. CONCLUSION: BSG is better than Zaizhang Shengxue Tablet in clinical effect and improving the CAA related adhesive molecules, Fn and FL, suggesting that it is an effective remedy for preventing and treating CAA, with its curative effect more significant on patients of yang-deficiency type than that on patients of yin-deficiency type.

[Influence of packed red cell transfusions on blood selenium concentration in pediatric hemato-oncology]. / Einfluss der Transfusion von Erythrozytenkonzentrat auf die Selenkonzentration im Vollblut bei Kindern mit hämato-onkologischen Erkrankungen.

BACKGROUND: In humans approx. 10% of the total body selenium (Se) content is present in the blood being evenly distributed among plasma and red cells. The important role of Se in antioxidative biological pathways is proven. Many parents of children with malignancies ask for supplementation with Se as part of complementary therapy during or after the oncological treatment. However, toxic Se concentrations may easily be reached in children. In order to analyse whether Se is also supplied by red cell transfusions (RCT), we determined Se concentration in whole blood prior and after packed RCT in pediatric patients with hemato-oncological diseases. PATIENTS AND METHODS: EDTA-blood was collected from 17 patients (median age: 4 years, range: 1 month - 17 years) with aplastic anemia, acute leukemia and solid tumours prior and after RCT (n=60). Patients received a median of 2 transfusions (range: 1-14). Samples were also collected from the transfusion blood bags and Se concentration was determined quantitatively by atomic absorption spectrometry. RESULTS: 95% of the specimen collected from the transfusion bags exhibited selenium concentrations within the normal adult range. Mean Se concentration in the patients' blood prior to RCT was 66.2 microg/l (range: 38.0-166.4 microg/l) and increased to 70.7 microg/l (range: 14.1-105.1 microg/l) thereafter (statistically not significant). Applying age dependant reference values Se concentrations were below the lower limit in 45% of the samples prior to RCT and only in 26% after RCT. The reason for this increase was the fact that Se concentrations were often just marginally below the age-dependant lower limit prior to RCT and in the lower normal range thereafter. CONCLUSION: 43% of the patients with hemato-oncological diseases in this study exhibited no Se deficiency at any time point. In the remaining 57% of the patients a transient or persistent Se deficiency was detected with blood levels partially far below the lower threshold of the age adjusted normal range. The Se deficiency was corrected in four out of eight patients by RCT. As Se levels may fluctuate in individual pts a supplementation should only be initiated if based on regular monitoring of the Se concentration.

Clinical study on effect of Astragalus Injection and its immuno-regulation action in treating chronic aplastic anemia.

OBJECTIVE: To observe the clinical effect of Astragalus Injection (, AI) and its immuno-regulatory action in treating chronic aplastic anemia (CAA). METHODS: Sixty patients with CAA were randomly assigned to two groups equally, both were treated with Stanozolol three times a day, 2 mg each time through oral intake, but AI was given additionally to the patients in the treated group once a day via intravenous dripping. All were treated for 15 days as one therapeutic course and the whole medication lasted for more than 4 months totally, with follow-up adopted. The clinical efficacy was estimated and the changes of T-lymphocyte subsets in peripheral blood as well as the serum levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2) were observed. RESULTS: The total effective rate in the treated group was 83.3% (25/30), which was higher than that in the control group 66.7% (20/30), showing significant difference between them (P<0.05). Levels of hemoglobin, WBC, reticular cell and platelet were elevated in both groups after treatment, but the improvement was significantly better in the treated group than that in the control group with respect to the former three indexes (P<0.05). The level of CD4(+) increased and that of CD8(+) decreased significantly after treatment in the treated group (P<0.05), which showed significant difference as compared with those in the control group (P<0.05). Levels of serum TNF-alpha and IL-2 lowered after treatment in both groups, but significance only showed in the treated group (P<0.05). The degree of proliferation in bone marrow got raised significantly and the percentage of non-hemopoietic cells reduced significantly in the treated group after treatment, also showing significant difference to those in the control group (P<0.05). CONCLUSION: AI could promote the recovery of hemopoietic function, which might be through improving T-lymphocyte subsets and reducing the release of negative regulatory factors such as TNF-alpha and IL-2 to alleviate the inhibition on hemopoietic function.

[Effects of Fuzhengyangying granules on the bone marrow proliferation and bcl-2 expression in mice with immune mediated aplastic anemia].

OBJECTIVE: To explore the effect of Fuzhengyangying granules (FZYYG) on the expression of the bone marrow proliferation and bcl-2 in mice with immune mediated aplastic anemia. METHODS: Forty BALB/c mice were randomly allocated to 4 groups: a normal control group, a model group, cyclosporine A (CSA) group and FZYYG group. The model group was fed with physical salt, while the CSA and FZYYG group were fed with CSA and FZYYG respectively, and then the changes of peripheral hemoglobin (Hb), platelet, bone marrow nucleus cell (BMNC), bone marrow hematopoiesis tissue volume and bcl-2 expression were examined. RESULTS: The count of Hb, the platelet, BMNC, the bone marrow hematopoiesis tissue volume and the bcl-2 positive rate in the model group were lower than those in the normal group (P<0.05), whereas the count of Hb, the platelet, BMNC, the bone marrow hematopoiesis tissue volume and the bcl-2 positive rate in both CAS and FZYYG groups were significantly higher than those in the model group (P<0.05). CONCLUSION: FZYYG can induce the bcl-2 antigen expression and postpone the haematogenous cells apoptosis, and it is effective for mice with immune mediated aplastic anemia.

Altered functional status of the hypothalamic dopaminergic tone in patients with chronic graft-versus-host disease after allogeneic hematopoietic stem cell transplantation: a pilot study.

We compared the functional status of the hypothalamic dopaminergic tone in patients given an allogeneic hematopoietic stem cell transplantation (allo-HSCT) with chronic graft-versus-host disease (GVHD) with that observed in patients with allo-HSCT without chronic GVHD and in healthy controls. The effect of acute dopaminergic blockade with intravenous metoclopramide on serum prolactin (PRL) concentrations was evaluated. Twenty volunteers, 20 to 52 years of age, seronegative for both hepatitis C virus and the human immunodeficiency virus, were studied: (1) 10 clinically healthy men (group 1), and (2) 9 patients with leukemia, and 1 patient with refractory aplastic anemia who underwent allo-HSCT, 5 of whom (3 men and 2 women) developed chronic GVHD (group 2), and 5 (3 men and 2 women) who did not develop chronic GVHD (group 3). Serum PRL concentrations were measured both fasting and after intravenous administration of metoclopramide (10-mg bolus). The area under the PRL curve was calculated. Patients in group 2 were older than those in groups 1 and 3 (P<.018), but their body mass index was similar. Fasting serum PRL concentrations were similar among the 3 groups; however, group 2 had higher PRL concentrations throughout the test (P<.001) and a greater area under the PRL curve than groups 1 and 3 (P<.001), without differences between the last 2 groups. The differences remained significant after adjustment for age (P<.01). Our results in a small group of patients with chronic GVHD after allo-HSCT suggest the existence of an increased functional level of their hypothalamic dopamine tone, which would favor a tendency toward a diminished endogenous production, release of pituitary PRL, or both. This could represent an adaptive mechanism aiming to maintain circulating PRL concentrations within a physiological range.

[Clinical efficacy and T-lymphocyte subset, serum interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2(IL-2) levels on treatment of chronic aplastic anemia patients by shenfu injection combined with stanozol and cyclosporin A].

OBJECTIVE: To observe the effect of Shenfu injection (SFI) and influence on T-lymphocyte subset, serum level of interferon-gamma(IFN-gamma), tumor necrosis factor-alpha(TNF-alpha), interleukin-2(IL-2) in patients with chronic aplastic anemia (CAA) based on treating with stanozol and cyclosporin A. METHOD: 60 patients with CAA were randomly divided into two groups, 30 patients in the SFI group were treated with SFI (100 mL which contains Ginsenoside 0.8 mg x mL(-1) and aconitine 1.8 microg x mL(-1) by adding it in 500 mL of 5% glucose every day) plus stanozol and cyclosporin A and 30 patients in the control group treated with slanozol and cyclosporin A alone for 2 months. The clinical efficacy was observed. The change of T-lymphocyte subset analyzed by flow cytometry and the levels of serum IFN-gamma, TNF-alpha, IL-2 measured with ELISA method were also observed before and after treatment. RESULT: After treatment, the total effective rate of the SFI group was higher than that in the control group, but it did not showing significant difference. The CD4/CD8 levels were significantly increased (1.76+/-0.49, P< 0.01) and CD8 levels were significantly lowered (22.57+/-6.30, P < 0.01) in the SFI group after treatment. Serum levels of lFN-gamma, TNF-alpha and IL-2 were lower in both groups, and the level of TNF-alpha and IL-2 in the SFI group (0.710+/-0.213) ng x L(-1) and (0.639+/-0.247) ng x L(-1) was significantly lowered than that in the control group (P < 0.05, P < 0.01). CONCLUSION: SFI might believe the hemopoietic inhibition so as to promote the recovery of hemopoietic function through improving the T-lymphocyte subset and reducing the release of hemopoietic negative regulatory factors such as IFN-gamma, TNF-alpha and IL-2.

[Effect of Panax notoginsenosides on the proliferation of hematopoietic progenitor cells in mice with immune-mediated aplastic anemia].

OBJECTIVE: To study the effect of panax notoginsenosides (PNS) on the proliferation of hematopoietic progenitor cells (HPC) in mice with immune-mediated aplastic anemia. METHODS: Balb/c mice model of immune-mediated aplastic anemia was established by radiation with sublethal dose of 60Co following the intravenously infusing lymphocytes of DBA/2 mice. Model mice in the treated groups were treated separately with high, middle and low dose of PNS, 3.2 mg, 1.6 mg and 0.8 mg per day respectively by intraperitoneal injection. Model mice in the control group and normal mice in the normal control group were treated with normal saline. The peripheral white blood cell (WBC) count and pathological examination of bone marrow were carried out 12 days later, the bone marrow was taken to be incubated in semi-solid culture system for observing proliferation of HPC. RESULTS: PNS could (1) increase peripheral WBC count: as compared with that in the model control, WBC in the high, middle and low dose PNS groups was raised by (34.3 +/- 2.9)%, (29.2 +/- 1.7)% and (14.5 +/- 1.6)% respectively, P < 0.01 and P < 0.05; (2) improve the bone marrow inhibition: pathological examination showed in the model group, the hematopoietic structure was destroyed and replaced by fatty tissue, while in the PNS treated groups, the structure of marrow was rather complete and filled with abundant hematopoietic cells; (3) promote the proliferation of HPC: as compared with the model group, the colony formation of CFU-GM were increased by (64.4 +/- 2.8)%, (67.3 +/- 2.4)% and (21.9 +/- 1.8)% respectively and that of CFU-E increased by (31.9 +/- 3.6)%, (20.7 +/- 2.4)% and (12.8 +/- 2.6)% respectively in the three PNS treated group (P < 0.01 and P < 0.05). CONCLUSION: PNS could enhance hematopoiesis by promoting proliferation of CFU-GM and CFU-E progenitors so as to improve the hematopoietic function in mice of immune-mediated aplastic anemia.