Immune thrombocytopenia purpura (ITP) is a disorder mediated by antiplatelet antibodies and characterized by accelerated destruction of platelets and impaired platelet production. The mainstay therapies for ITP have included corticosteroids, the immune globulin intravenous immunoglobulin and IV anti-D (monoclonal antibodies against the D antigen of the Rh system), vinblastine or a monoclonal anti-CD20 antibody that transiently depletes CD20(+) B cells, danazol, cyclophosphamide and even splenectomy to refractory one. Most of ITP patients responded to those treatment, while more than 30% of whom may relapse sooner or later. The recombinant forms of human TPO were discontinued from human use in clinic because recipients of these agents developed significant thrombocytopenia secondary to production of neutralizing antibodies that cross-reacted with endogenous TPO. All above mentioned treatments have side effects and severe infection may arise post splenectomy. The more powerful treatment with less side effects are needed. There are two TPO receptor agonist, AMG531 and Eltrompobag, have approved in Europe for the treatment of ITP. Both of them can improve the differentiation of megakaryocyte and platelets production. Combination treatment including pancytoprotector shows good effect in the treatment of refractory and relapsed ITP in China. Altogether, individual treatment of ITP is the contemporary trend in both clinical and preclinical practice. In this review the pathogenesis of ITP and its clinical therapies were reviewed, the individual regiments for treating ITP patients were discussed.
Humans ; Purpura, Thrombocytopenic, Idiopathic ; classification ; immunology ; therapy

ObjectiveTo observe the influence of different dietary protein intake (DPI) on nitrogen balance and nutritional indices in peritoneal dialysis (PD) patients, and explore the minimal DPI to maintain nitrogen balance.MethodsThirty-four PD patients were randomly divided into group A, B and C with DPI as 1.2, 0.9 and 0.6 g·kg-1·d-1 respectively. All the patients admitted into our hospital and completed a 10-day assessment for nitrogen balance, as well as nutritional status including serum albumin (Alb), pre-albumin at baseline, the 7th and 10th day. ResultsThe DPI of group A, B and C was (1.18±0.05), (0.87±0.02), (0.66±0.03) g·kg-1·d-1, whose differences were significant (P<0.01). The dietary energy intake (DEI) was 129.29 (117.57-133.89), 111.71 (100.42-133.47), 146.86 (128.03-163.18) kJ·kg-1·d-1 respectively. Nitrogen balance was positive in group A, B, C [2.99 (2.15-4.72) g, 1.20(0.59-1.89) g, 0.24 (-0.87-1.27) g]. The BUN decreased at the 7th and 10th day (P<0.01) in group C. The BUN and phosphorus in group A increased, but without significant difference as compared to baseline. No significant differences of nutritional status were found among three groups throughout the trial. ConclusionMinimal DPI 0.65 g·kg-1·d-1 plus the supplement of protein loss in dialysate can maintain the nitrogen balance in peritoneal dialysis patients.

OBJECTIVETo investigate the effect of miRNA silencing HIF-1α gene on the proliferation of HepG2 cells.

METHODSThe eukaryotic expression plasmids of HIF-1α miRNA and report gene containing hypoxia-reponse element were constructed and transfected into HepG2 cells. The expressions of HIF-1α gene and protein were determined by real time-PCR and Western blotting. The expressions of HIF-1α, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) were quantitatively detected by ELISA. The alterations of cell cycles and apoptosis rate were quantitatively measured by flow cytometry and Annexin V-FITC/PI double dyeing assay.

RESULTS72 h after transfection the down regulations of HIF-1α mRNA and protein were 87% and 56% respectively, and the decrease of target gene was 46% in the report gene, 54% in VEGF and 36% in Ang-2, respectively. The apoptotic ratio of HepG2 cells was 22.46+/-0.61% (P < 0.01). The cell cycle changed greatly at the ratio of G1 (61.49+/-1.12%) and S (22.40+/-0.58%, P < 0.01). After being combined with doxorubicin, the apoptotic ratio increased to 36.99+/-0.88% and the ratios of G1 and S phases were upregulated to 65.68+/-0.91% and 19.47+/-1.34% respectively.

CONCLUSIONSHIF-1α miRNA or / and doxorubicin can regulate the growth cycles of HepG2 cells, promote the cell apoptosis and inhibit the cell proliferation.

Apoptosis ; Cell Cycle ; Cell Proliferation ; Gene Silencing ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; MicroRNAs ; genetics ; RNA, Messenger ; genetics ; Transfection

The objective of this study was to explore the effect of amifostine (AMF) in treating patients with idiopathic thrombocytopenic purpura (ITP) and its adverse effects. 17 ITP patients (aged from 13 to 92, mean 65 years) were treated with AMF and their therapeutic regimen was following: 0.4 g AMF was dissolved in 100 ml physiological saline, the intravenous drop was performed at day and lasted 5 days per week, with interval 2 days, 4 weeks were considered as a course of treatment. The results showed that the normal counts of platelets were observed in 17 ITP patients after 1 course of treatment, and remained unchanged for 2 months following stop of therapy. In that time, no any drugs such as hormonal preparation, gamma-globulin, blood transfusion and so on were given in all treated patients, the main adverse effect of AMF was discomfort in digestive system that all patients could endure. Therefore, the patient's life quality was obviously improved. It is concluded that the AMF has been initially and successfully used to treat ITP patients with positive response, and may be considered as a safe-effective drug for treating old patients with ITP, but the long-term effect and pharmacological mechanism of this drug needs further evaluation.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Amifostine ; adverse effects ; therapeutic use ; Female ; Humans ; Infusions, Intravenous ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

The objective of the study is to explore the effect of Fas, FasL and Bcl-2 on the process of apoptosis induced by chemotherapeutic drugs through detecting the expression of Fas, FasL and Bcl-2 on murine lymphoma cell line RMA. Dexamethasone(DEX), etoposide (VP-16), arsenic trioxide As(2)O(3) and all trans-retinoic-acid (ATRA) were added to the RMA cells as well as to the cells preincubated with interleukin-2 (IL-2), interleukin-6 (IL-6) or granulocyte-macrophage colony-stimulating factor (GM-CSF), respectively. The effect on apoptosis was observed and the expression of Fas and FasL mRNA as well as the expression of Fas and Bcl-2 antigen were measured. DEX and VP-16 could promote apoptosis of RMA cells while upregulating the expression of Fas and FasL without affecting the expression of Bcl-2. ATRA downregulated the expression of Bcl-2 without any change of Fas and FasL, and no apoptosis of RMA cells induced by ATRA was observed. Although As(2)O(3) induced apoptosis of RMA cells, it did not affect the expression of Fas, FasL and Bcl-2, which suggested that different drugs induce apoptosis of the same kind of cells by different signal transduction system and apoptosis induced by Fas system needed the coexistence of Fas and FasL. Although IL-2, IL-6 and GM-CSF upregulated the expression of Fas protein when adding to RMA cells separately, none of them induced apoptosis. Apoptosis could be induced by combination of IL-2 and IL-6 along with the upregulation of Fas and FasL. The cytokines facilitated the apoptotic action of chemotherapeutic drugs, the drug concentration for inducing apoptosis decreased and the time period of starting apoptosis shortened. Apoptosis could be observed without the expression of FasL when anti-Fas-antibody was added to RMA cells. The results demonstrated that there was synergistic effect of chemotherapeutic drugs and some cytokines for induction of apoptosis. Fas-FasL system participated in the apoptosis induced by DEX and VP-16; different drugs induce apoptosis by different pathway of signal transduction.
Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Apoptosis ; physiology ; Arsenicals ; pharmacology ; Dexamethasone ; pharmacology ; Etoposide ; pharmacology ; Fas Ligand Protein ; Gene Expression ; drug effects ; Membrane Glycoproteins ; biosynthesis ; Mice ; Oxides ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Tretinoin ; pharmacology ; Tumor Cells, Cultured ; fas Receptor ; biosynthesis

CYP2D6 is an important drug-metabolizing enzyme. The polymorphism of CYP2D6 leads to metabolism difference and the different reactions of drugs in the individuals and different races are normal phenomenon in clinical medication. CYP2D6*10 is an important subtype in Asian people and 51.3% Chinese are classified with this subtype. To obtain recombinant active CYP2D6*1/CYP2D6*10 in baculovirus system by optimizing coexpression with CYPOR, and detect their activity to catalyze dextromethorphan, three recombinants pFastBac-CYP2D6*1, pFastBac-CYP2D6*10 and pFastBac-CYPOR were constructed and transformed into DH10Bac cell to obtain the recombinant Bacmid-CYPOR, Bacmid-CYP2D6*1 and Bacmid-CYP2D6*10. And then the recombinant CYP2D6*1 and CYP2D6*10 virus were obtained by transfecting Sf9. Then homogenate protein activity was determined with dextromethorphan as substrate. The multiple of infection (MOI) and its ratio of recombinant CYP2D6 virus to CYPOR virus were adjusted by detecting the activity of the homogenate protein. The Km and Vmax are 26.67 +/- 2.71 micromol x L(-1) (n=3) and 666.7 +/- 56.78 pmol x nmol(-1) (CYP2D6) x min(-1) (n=3) for CYP2D6*1 to catalyze dextromethaphan. The Km and Vmax are 111.36 +/- 10.89 micromol x L(-1) (n=3) and 222.2 +/- 20.12 pmol x nmol(-1) (CYP2D6) x min(-1) (n=3) for CYP2D6*10 to catalyze dextromethorphan. There is significant difference between CYP2D6*1 and CYP2D6*10 for Vmax and Km (P < 0.01). The clearance ratio of CYP2D6*1 is 25.0 and the clearance ratio of CYP2D6*10 is 2.0. The expressed CYP2D6*1 and CYP2D6*10 are useful tools to screen the metabolism profile of many xenobiotics and endobiotics in vitro, which are benefit to understand individual metabolism difference.
Animals ; Baculoviridae ; enzymology ; genetics ; Catalysis ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Cytochrome P-450 CYP2D6 ; genetics ; metabolism ; Dextromethorphan ; metabolism ; Isoenzymes ; metabolism ; NADPH-Ferrihemoprotein Reductase ; genetics ; metabolism ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Spectrometry, Mass, Electrospray Ionization ; Spodoptera ; cytology ; virology ; Transfection

The aim of this study was to investigate the curative effect of amifostine (AMF) combined with recombinant human erythropoietin (rhEPO) on the aged patients with myelodysplastic syndrome. Two aged MDS patients (one aged 91; another 86) were treated with amifostine and rhEPO over a period of 4 weeks. The results showed that a short-term curative effect was observed and transfusion interval was prolonged in both patients after 4 week treatment with 5 x 0.4 g AMF plus 3 x 6,000 U rhEPO per week. The reticulocyte count in MDS-RA patient returned to normal at first week of treatment and still remained in normal level for 4 weeks; leukocyte, hemoglobin and platelet values in peripheral blood of MDS-RCMD patient obviousby increased, the abnormally increased reticulocyte value displayed a decrease trend after amifostine plus rh-EPO treatment. In conclusion, amifostine plus rhEPO may have a good therapeutic effect for aged MDS patients, and its clinical long-term curative effect still needs further evaluation.
Aged ; Aged, 80 and over ; Amifostine ; therapeutic use ; Drug Therapy, Combination ; Erythropoietin ; therapeutic use ; Humans ; Male ; Myelodysplastic Syndromes ; drug therapy ; Recombinant Proteins ; Time Factors ; Treatment Outcome

The present study was aimed to investigate the damage effect of hepatocyte growrh promoting substance (HGS) on the HL-60 cell DNA in vitro and to explore the possible mechanism underlying the effect. Experiment was divided into 3 groups: one was control group, in which 0.9% NaCl solution was added, and other two were experimental group 1 and group 2, where 22.5 microg/ml and 40 microg/ml HGS were added, respectively. HL-60 cell growths were compared between groups with and without HGS. Single cell gel electrophoresis (SCGE) was used to detect DNA damage of HL-60 cell. DNA electrophoresis was used to detect the apoptosis of HL-60 cells caused by HGS. The results showed that the inhibitory effects of HGS on growth of HL-60 cells were observed in group with 22.5 microg/ml and group with 40 microg/ml after culture for 2 days, the DNA ladder and the apoptosis of HL-60 cells occurred in these 2 groups on day 2 after addition of HGS, the counts of HL-60 cells with comet tail in these experimental groups were found to be more increased in comparison with control group. In conclusion, the HGS can inhibit the growth of HL-60 cell and the apoptosis of HL-60 cells should be induced through pathway of DNA damage caused by HGS.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; genetics ; Cattle ; Cell Proliferation ; drug effects ; Comet Assay ; DNA Damage ; DNA, Neoplasm ; analysis ; genetics ; HL-60 Cells ; Humans ; Peptides ; pharmacology

The purpose of this study was to explore the effect of amifostine in treating aged patients with idiopathic thrombocytopenic purpura (ITP). Three aged ITP patients (one aged 88, one aged 75, another aged 65) were treated with amifostine over a period of 4 weeks for each. The results showed that the early positive responses were observed in all 3 aged patients after treatment with amifostine 5 x 400 mg per week for 4 weeks, the recovery of platelet counts to normal level were found in 2 cases after 1 week and in 1 case after 2 weeks. Following 4 months after stop of treatment with amifostine, the normal platelet counts still remains in all patients, no more blood transfusions, hormonal preparation and gamma-globulin were given. Amifostine has never been reported elewhere as to treat of aged ITP. In conclusion, amifostine has a good early therapeutic effect on aged ITP patients, but its clinical outcome of long-term still needs further evaluation.
Aged ; Aged, 80 and over ; Amifostine ; therapeutic use ; Drug Administration Schedule ; Humans ; Male ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; Treatment Outcome

One case of hairy cell leukemia with survival for 10 years was reported. The patient received Interferon-alpha(2b) treatment continuously for more than 9 years and intravenous administration of fludarabine for 2 courses of treatment in the later period, and died due to complication of severer infection in thd end. The manifestation, diagnosis and treatment of hairy cell leukemia were discussed.
Aged ; Humans ; Immunophenotyping ; Interferon-alpha ; therapeutic use ; Leukemia, Hairy Cell ; diagnosis ; drug therapy ; mortality ; Male ; Recombinant Proteins ; Vidarabine ; analogs & derivatives ; therapeutic use