Efficacy of intermittent iron supplementation in children with mild iron-deficiency anemia / 中国当代儿科杂志

OBJECTIVES@#To study the efficacy of intermittent iron supplementation in children with mild iron-deficiency anemia.@*METHODS@#A total of 147 children with mild iron-deficiency anemia were enrolled in this prospective study. They were divided into an intermittent iron supplementation group (n=83) and a conventional iron supplementation group (n=64). The levels of hemoglobin were measured before treatment and after 1 and 3 months of treatment. The treat response rate and the incidence rate of adverse drug reactions were compared between the two groups.@*RESULTS@#Both groups had a significant increase in the level of hemoglobin after iron supplementation (P<0.05). After 1 month of treatment, the conventional iron supplementation group had a significantly higher treatment response rate than the intermittent iron supplementation group (61% vs 42%, P<0.05). After 3 months of treatment, there was no significant difference in the treatment response between the two groups (86% vs 78%, P>0.05). The incidence rate of adverse drug reactions in the conventional iron supplementation group was significantly higher than that in the intermittent iron supplementation group (25% vs 8%, P<0.05).@*CONCLUSIONS@#For children with mild iron-deficiency anemia, although intermittent iron supplementation is inferior to conventional iron supplementation in the short-term efficacy, there is no significant difference in the long-term efficacy between the two methods, and compared with conventional iron supplementation, intermittent iron supplementation can reduce the incidence of adverse drug reactions, alleviate family financial burdens, and improve treatment compliance of children, thus holding promise for clinical application.

Mobilization Effect of HS6101 on Hematopoietic Cells of Mice / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the mobilization effect of HS6101 on hematopoietic cells of mice.</p><p><b>METHODS</b>The normal ICR mice were injected subcutaneously once or twice with HS6101 at 9 µg/d/mouse, or a single dose of HS6101 3, 9, 27 and 81 µg/mouse was administrated, and the mobilization effect of HS6101 in different administration times and different dosage was observed, and compared with the synergistic effects of administration of single dose of HS6101 combined with rhG-CSF (2 µg/d/mouse was injected subcutaneously for 5 consecutive days). The peripheral blood cell counts of mice were detected at different time after administration. The hematopoietic stem/progenitor cells of bone marrow and peripheral blood were detected at day 5 and 10 after administration.</p><p><b>RESULTS</b>There was no significant difference in peripheral blood cell counts after once or twice injections of HS6101 9 µg/mouse. The peripheral platelet counts dose-dependently increased in ICR mice, which accounted for 121.1% to 118.0%, 138.7% to 123.1%, 146.4% to 139.2%, and 156.2% to 168.7% (P < 0.001) after HS6101 (3, 9, 27 and 81 µg/mouse) treatments at 5 and 7 d, respectively. HS6101 (3, 9, 27 and 81 µg/mouse) showed dose-response relationship to platelets, with R value of 0.777 and 0.954 at day 5 and 7 after administration, respectively. HS6101 significantly increased numbers of hematopoietic stem/progenitor cells in both bone marrow and peripheral blood, and elevated peripheral blood leukocytes at 27 µg/mouse dose at day 5 after administration.</p><p><b>CONCLUSION</b>HS6101 has significant mobilization effect on hematopoietic stem/progenitor cells, platelets and leukocytes in mouse.</p>

Effects of HS 6101 and rhG-CSF on Hematopoiesis Recovery of ICR Mice Injured by Cyclophosphamide / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of HS 6101 and recombinant human granulocyte colony stimulating factor (rhG-CSF) on hematopoiesis recovery of ICR mice injured by cyclophosphamide (CTX).</p><p><b>METHODS</b>A total of 103 ICR mice were divided into 4 groups, including CTX control (46 mice), HS 6101 (21 mice), rhG-CSF (18 mice) and HS 6101+rhG-CSF (18 mice), respectively. The mouse model of chemotherapy-induced haematopoietic injury was established by CTX intraperitoneal injection at a dose of 100 mg/kg once a day for 3 consecutive days. Single dose of HS 6101 (27 µg/mouse) was injected subcutaneously at 1 hour before the first administration of CTX. One day after the last CTX treatment, 2 µg/mouse of rhG-CSF was injected subcutaneously once a day for 5 consecutive days. The peripheral blood cell counts of the mice were observed once every 2 days. Hematopoietic progenitor cell colony counting and histopathological assessment of bone marrow cells were evaluated in the mice at days 4 and 9 after the first administration of CTX.</p><p><b>RESULTS</b>Both HS 6101 and rhG-CSF alone or in combination, significantly elevated the nadirs of peripheral blood leukocytes and neutrophils, increased the number of bone marrow hematopoietic progenitor cells, and stimulated the hematopoietic cell hyperplasia of bone marrow in the mice treated with CTX. The effect of HS 6101 combined with rhG-CSF was better than that of the drugs used alone.</p><p><b>CONCLUSION</b>The HS 6101 at 27 µg/mouse can significantly promote the recovery of hematopoiesis in ICR mice treated with CTX chemotherapy, and its combination with rhG-CSF shows synergistic effects.</p>

Protective effects of WR2721 on early bone marrow hematopoietic function in mice exposed to 6.5 Gy of (60)Co γ-rays / 中国实验血液学杂志

The aim of this study was to investigate the effect of WR2721(amifostine) against bone marrow hematopoietic damage of mice exposed to 6.5 Gy of (60)Co-γ ray. A total of 60 C57/BL6J mice was divided into 3 groups:normal group (mice were injected with physiological salt solution), irradiation group (mice were injected with physiologic salt solution before irradiation) and WR2721 group (mice were injected with WR2721 before irradiation). The WBC, neutrophil (Neut), Plt and RBC levels in peripheral blood of 3 group mice were counted within 60 days after irradiation; the bone marrow nuclear cells (BMNC) were counted at 2 and 24 hours after irradiation; the hematopoietic stem/progenitor cell (LK/LSK) level and colony formation capability were detected by flow cytometry at 2 and 24 hours after irradiation. The results indicated that the counts of WBC and neut at 4 and 18 days, Plt at 7-18 days and RBC at 10-30 day after irradiation in WR2721 group were higher than those in irradiation group (P < 0.05); the BMNC, LSK and LK levels obviously increased at 24 hours after irradiation (P < 0.05), the CFU-GEMM, CFU-GM, CFU-MK BFU-E and CFU-E all significantly increased at 2 and 24 hours after irradiation (P < 0.01), as compared with irradiation group. It is concluded that WR2721 can effectively alleviate early hematopoietic damage and promote the fast recovery of peripheral blood cells in mice exposed to γ-ray, suggesting that the WR2721 has significant radioprotective effect on hematopoietic system.

Research progress on effect factors of secondary metabolites content in callus / 中国中药杂志

Secondary metabolites are the result of that plant interaction with biological and non-biological factors in the long-term evolution process, and play an important role in plant growth, development and physiology. The effective components of medicinal plant are usually the secondary metabolites in plant cells, and the synthesis of them are affected by a variety of factors, such as environmental impact. Acquirement of the secondary metabolites via callus culture has the advantage of low cost and less environmental impact. The synthesis and accumulation of medicinal plant secondary metabolites are not only controlled by light, temperature and pH, but also infected by germplasm, plant growth regulator and elicitor. This article presents a review of the influencing factors, and provides a basis for further study and development.

High dose granulocyte colony-stimulating factor enhances survival and hematopoietic reconstruction in canines irradiated by 2.3 Gy mixed fission neutron and gamma ray / 中国实验血液学杂志

This study was purposed to evaluate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on hematopoietic reconstruction and survival in beagles exposed to mixed fission neutron and γ-ray. 13 beagles were unilaterally exposed to single dose of 2.3 Gy 90% neutrons. The experiments were divided into 3 groups: irradiation control group (no any treatment, n = 4), supportive care group (n = 5) and rhG-CSF plus supportive care group (n = 4, abbreviated as rhG-CSF group) in which the beagles were subcutaneously injected with 200 µg/kg of rhG-CSF early at half an hour and 24 hours post-irradiation respectively. The results showed that 2.3 Gy 90% neutron irradiation induced a severe acute radiation sickness of bone marrow type. The administration of rhG-CSF increased the survival rate from 60% in supportive care group to 100%. Twice injection of rhG-CSF in the first 24 hours reduced duration of neutropenia, enhanced neutrophil nadir and promoted neutrophil recovery when compared with control cohort administered clinical support. The number of colony-forming cells (CFU-GM, CFU-E, and BFU-E) in peripheral blood of rhG-CSF treated canines increased 2-to 5-fold relative to those of the supportive care group on day 3. All canines treated with rhG-CSF achieved hematopoietic reconstruction as evidenced by the pathological section of sternum while severe shortage of hemopoietic cells remained in the cohorts given supportive care alone. It is concluded that the combination of supportive care and high-dose rhG-CSF can accelerate hematopoietic recovery and enhance survival of dogs exposed to 2.3 Gy mixed neutron and gamma ray.

Effect of rhG-CSF on blood coagulation in beagles irradiated by 2.3 Gy neutron / 中国实验血液学杂志

The aim of this study was to investigate the effect of recombinant human granulocyte stimulating factor (rhG-CSF) on blood coagulation of beagles irradiated by 2.3 Gy neutron so as to provide new therapy for blood coagulation disorder after neutron irradiation. 10 beagles were exposed to 2.3 Gy neutron, and then randomly assigned into supportive care group and rhG-CSF-treated group. The rhG-CSF-treated cohorts were injected subcutaneously with rhG-CSF (10 µg/kg·d) beginning at the day of exposure for 21 consecutive days. Peripheral blood platelet counts were examined once every two days. In vitro platelet aggregation test, thromboelastography and blood clotting tetrachoric tests were also performed. The results indicated that the blood clotting system of irradiated dogs was in hypercoagulable state in the early days after 2.3 Gy neutron irradiation, and became hypocoagulable at crisis later and were mainly on intrinsic coagulation pathway. Blood fibrinogen increased markedly during the course of disease, while platelet counts and aggregation function were decreased remarkably. rhG-CSF administered daily could correct hypercoagulable state induced by 2.3 Gy neutron irradiation at the early time post exposure, shortened the thromboplastin generation time and clotting formation, down-regulated the abnormal high fibrinogen in blood, and improved platelet aggregation function. It is concluded that rhG-CSF can improve coagulation disorders of irradiated dogs.

Preparation and identification of monoclonal antibody against enoyl-CoA hydratase 1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (mAbs) against enoyl-CoA hydratase 1 (ECH1).</p><p><b>METHODS</b>Normal human liver tissues were homogenized, and the mitochondria were isolated by differential centrifugation. The total mitochondrial proteins were used to immunize BALB/c mice to prepare mAbs by routine hybridoma technique. The mAbs were characterized by ELISA, Western blotting and immunohistochemistry. The specificity of the antibody was identified by mass spectrometry (MS) following immunoprecipitation (IP) and confirmed by Uni-ZAP expression library screening.</p><p><b>RESULTS</b>One clone of the hybridoma BGB095 secreting specific mAb against ECH1 was obtained. The mAb was identified to belong to Ig subclass IgG1 and could be used in ELISA, Western blotting, immunohistochemistry, and immunoprecipitation.</p><p><b>CONCLUSION</b>A hybridoma cell line stably secreting specific mAb against ECH1 has been established. The specific mAb against ECH1 can be of great value for functional and distribution studies of ECH1.</p>

Effects and mechanism of CD4+ CD25+ regulatory T cells in mouse experimental colitis treated by CLYSTER No. 1 / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effects and mechanism of CD4+ CD25+ regulatory T cells (Tregs) in mouse experimental colitis treated by CLYSTER No. 1.</p><p><b>METHOD</b>The mouse model of experimental colitis was established by dinitrochlorobenzene (DNCB)-acetic acid (AA) in mice DNCB and AA. Adult KM mouse were randomly divided into four groups: normal control group, experimental colitis model group, SASP and Chinese medicine therapeutic groups. Proportion of CD4 CD25+ Tregs in peripheral blood (PB) and mesenteric lymph node (MLN) was estimated by flow cytometry at the end of one or two week after treating with SASP and CLYSTER No. 1.</p><p><b>RESULT</b>The model of experimental colitis in mouse was successfully established. Compared with normal control group, the proportion of CD4 CD25 Tregs was markedly decreased in PB and MLN of model control group of experimental colitis. But it was significantly increased in therapeutic groups of SASP and CLYSTER No. 1, and their CD4+ CD25+ Tregs in PB and MLN were much more than the model control group at the end of one or two weeks after treating with SASP and CLYSTER No. 1.</p><p><b>CONCLUSION</b>CD4+ CD25+ Tregs with strong immune suppression could play a central role in the initiation and development of mouse experiment colitis, and the CLYSTER No. 1 might exert its therapeutic effects on UC by the regulation of number and function of CD4+ CD25+ Tregs.</p>

Preparation and identification of monoclonal antibody against UGP2 / 中国实验血液学杂志

The study was aimed to generate monoclonal antibodies (mAbs) against homo sapiens UDP-glucose pyrophosphorylase 2 (UGP2). Normal human liver tissues homogenized, and cytosolic proteins isolated by centrifugation were used to immunize BALB/c mice to generate mAbs by hybridoma technique. The mAbs were identified by ELISA, Western blot, and immunohistochemistry assay. The antibody specificity was confirmed by Uni-ZAP expression library screening. The results indicated that one hybridoma BAD062 secreting specific mAb against UGP2 was established. The Ig subclass of this mAb was IgG(2b) (kappa), and it could be used in ELISA, Western blot, immunohistochemistry assay. The antigen recognized by BAD062 mAb was localized in the hepatocyte cytoplasm, with molecular weight of 56 kD in the cytosolic proteins of human liver tissue. The BAD062 mAb was further confirmed by immunoscreening of Uni-ZAP XR liver cDNA expression library. It is concluded that a hybridoma cell line stably secretes specific mAb against UGP2. This mAb reacted with UGP2 in ELISA, Western blot, immunohistochemistry assay, and would be very useful for the UGP2 studies.

Preparation and identification of monoclonal antibody against Homo sapiens hemoglobin alpha 2 (HBA2) / 中国实验血液学杂志

This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.

Recombinant human VEGF-D induces the angiogenesis of the chick embryo chorioallantoic membrane / 中国实验血液学杂志

Vascular endothelial growth factor-D (VEGF-D) and vascular endothelial growth factor receptor-2, -3 (VEGFR-2, -3) with their corresponding signaling pathway play significant roles in the development of the embryonic vascular system and pathological lymphangiogenesis. The study was aimed to express and purify the GST-VEGF-D fusion protein, and to explore the angiogenesis effect of VEGF-D. The total RNA was extracted from human fetal lung tissue, and the mature form of VEGF-D was expanded by polymerase chain reaction (PCR), then the plasmid pGEX-5X-1/VEGF-D was reconstructed and the GST-VEGF-D fusion protein expressed in transformed E.coli BL21-DE3. The results showed that the molecular mass of this fusion protein was 38 kD and compassed more than 15% of the total bacteria proteins. The fusion protein was recognized by anti-GST and anti-VEGF-D antibodies. The soluble GST-VEGF-D fusion protein could interact with VEGFR-3/Fc and was able to stimulate the proliferation of human erythroleukemia cell line (HEL) cells. The data of chick embryo chorioallantoic membrane (CAM) experiments indicated that GST-VEGF-D could induce the CAM angiogenesis. It is concluded that the GST-VEGF-D fusion protein with biological activity was successfully expressed, and which may provide an experimental model for the investigation of the VEGF-D-induced angiogenesis and lymphangiogenesis.

Characteristics of a new fibrinolytic enzyme produced from Rhizopus chinensis 12# / 生物工程学报

As a therapeutic agent in thrombosis the fibrinolytic enzymes are of interest and the search for a new enzyme continues. A novel fibrinolytic enzyme was produced from Rhizopus chinensis 120, which was screened from the starter for brewing rice wine in the South of China, by solid fermentation, and purified through ammonium sulfate precipitation, hydrophobic interaction, ionic exchange and gel filtration chromatographies. The purified enzyme hydrolyzed fibrin, it cleaved the alpha-, beta- and gamma-chains of fibrinogen simultaneously, and it also activated plasminogen to plasmin. The enzyme hydrolyzed N-Succinyl-Ala-Ala- Pro-Phe-pNA, and Km was 0.23 mmol/L and Kcat 16.36 s(-1). The optimal temperature of the enzyme for hydrolying fibrin was 45 degrees C, and the optimal pH range of 6.8 - 8.8. The isoelectric point of the enzyme estimated by isoelectric focusing electrophoresis was 8.5 +/- 0.1. The enzyme was a glycoprotein. EDTA, PCMB, PMSF inhibited the activety of the enzyme, and SBTI, Lys, TPCK, Aprotinine had none obvious inhibition, which suggested that the activity centre of the enzyme had hydrosulfuryl, metal and serine. The first 12 amino acids of the N-termimal sequence of the enzyme were NH2-Ser-Val-Ser-Glu-Ile-Gln-Leu-Met-His-Asn-Leu-Gly, and had none homology with that of other fibrinolytic enzyme from other microbes. The novel fibrinolytic enzyme from Rhizopus chinensis 12# has potential to become a therapeutic agent in thrombosis.

A brief report on effect of rhG-CSF in treating leukopenia after radio-and chemo-therapy of patients with breast cancer / 中国实验血液学杂志

The study was to observe the effect of rhG-CSF (lishengsu) in treating leukopenia caused by radiotherapy and chemotherapy in patients with breast cancer. 100 cases of breast cancer received modified radical mastectomy were randomized into two groups with the same treatment of one cycle chemotherapy using the protocol of CAF at two weeks after the operations and then radiotherapy. The patients in treated group received rhG-CSF 75 micro g per day s.c. for 5 - 7 days constantly, and additional 3 - 5 days according to leukopenia during radiotherapy. The patients in control group did not receive rhG-CSF during the chemo- and radio-therapy. The results shows that nadir of WBC and neutrophil counts in the treated group was higher than that in control significantly. In conclusion, effect of lishengsu on leucopenia in process of chemotherapy and radiotherapy shows definite therapeutic effect, the side effects are not remarkable.

The effects of rhG-CSF and rhSCF on peripheral blood leukocytes and CFU-GM in rhesus monkeys / 中国实验血液学杂志

To evaluate the effects of rhG-CSF and rhSCF on mobilization of the peripheral blood stem cells, 15 monkeys were divided into control, rhG-CSF 10 micro g/(kg x day) and rhG-CSF 10 micro g/(kg x day) + rhSCF 50 micro g/(kg x day) treated groups. Monkeys were administered with vehicle, rhG-CSF and rhG-CSF + rhSCF subcutaneously once daily for 14 days, respectively. The results showed that the highest counts of leukocyte of rhG-CSF treated group were 411% of baseline value on day 7 after administration, compared with that of rhG-CSF + rhSCF treated group which were 538% on day 9. The highest counts of leukocytes lasted for 3 days in combined treated group. CFU-GM from peripheral blood in the two groups were 8.37 and 11.75 times higher at 5 and 9 days respectively after the mobilization. It is concluded that rhG-CSF significantly increases the number of peripheral blood leukocytes and CFU-GM, and a better effect can be obtained by rhSCF + rhG-CSF combined administration.

Direct Injection of Plasmid DNA Expressing IL-6 Gene Improves Recovery of Thrombocytopoiesis in Irradiated Mice / 中国实验血液学杂志

Hemorrhage is one of major clinical features of the patients exposed to large dose of ionizing radiation and a sudden decrease of peripheral platelet counts in hemorrhage complication may bring the patients into life-threatening situation. Cytokines had been used to improve thrombocytopoiesis in various radiation induced thrombocytopenia. Current measures for this purpose involve repeated injection of recombinant cytokines, which bring much inconvenient and agony to the patients, or gene therapy with viral vectors that could not obviate the risk of infection. This work tried to determine the possibility of gene therapy with plasmid vectors for radiation-induced hematopoietic injury. After a single intramuscular injection of plasmid hIL-6 cDNA on 6.5 Gy irradiated mice, the IL-6 level began to increase from the day 4, reached the peak value about the day 11 and maintained at a higher level on the day 28, but the hIL-6 level showed less changes in unirradiated mice. Further experiments demonstrated the IL-6 level in 7.5 Gy irradiated mice was about three times higher than that of 5.0 Gy irradiated mice and the expression of hIL-6 in vivo showed significant effect on hematopoietic recovery. Not only the platelet nadir in peripheral blood, but also the number of colony-forming cells in bone marrow rose. It is concluded that radiation could significantly enhance the gene transfer efficiency of plasmid DNA and gene therapy with plasmid vectors for treating radiation-induced hematopoietic injury might be more effective than other diseases without DNA repair.

Therapeutic Effects of rhIL-11 on Irradiation-Induced Myelosuppression in Rhesus Monkeys / 中国实验血液学杂志

The efficacy of rhIL-11 in treating thrombocytopenia and neutropenia in gamma-irradiated rhesus monkeys and the variation in curative effect due to difference of administration times were studied. Healthy rhesus monkeys were exposed to 3.0 Gy (60)Co total body irradiation (TBI) to result in pancytopenia for three weeks. Treatment with rhIL-11 (30, 60 or 120 micro g.kg(-1).day(-1)) on early days (days 0 - 13 after TBI) could significantly improve the nadir of platelet count. Although the nadir of leukocyte count was not improved, the duration below 50% of its baseline value was shortened similarly to that of platelet. During the first two weeks after TBI, erythrocyte numbers of the animals treated with these doses of rhIL-11 were lower than those of the control group at first but they became higher beginning from the third week. Four monkeys were treated with rhIL-11 at 60 micro g.kg(-1).day(-1) on days 13 - 26 after TBI. The numbers of their peripheral blood cells followed the similar decrease patterns as those of control group during the first three weeks, then they were improved rapidly. By semi-solid bone marrow cell culture it was demonstrated that rhIL-11 could stimulate bone marrow cells to form more CFU-Meg, CFU-Mix, CFU-E, BFU-E and CFU-GM in vitro. Histopathological observation revealed that bone marrow of the control group was devoid of hematopoietic cells and bleeding, being contrary to that of the group treated with rhIL-11, in which the cells proliferated actively. The results suggest that rhIL-11 can accelerate hematopoietic recovery of irradiated monkeys.

A STUDY ON BIOTECHNOLOGY OF GLYCOLIPID PRODUCED BY A BACILLUS COAGULANS / 微生物学通报

The biotechnology of glycolipid fermented b y a bacillus coagulans was studied and the fermentation pro cess in 10L bioreactor was conducted.Suitable medium contained 6% bean oil as ca rbon source,3 5g/L NaNO 3 as nitrogen source,0 75g/L yeast extract and some i no rganic salts.The fermentation temperature of 30℃,initial pH of 8 5,strring rev olution of 150～240r/min and fermentation period of 96h proved to be optimal.The yield of glycolipid in 10L bioreactor was 7 073g/L.