Adult ; Comet Assay ; DNA Damage ; drug effects ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Peracetic Acid ; toxicity

To explore the relationship between the expression of CD133 and pathogenesis of leukemia and MDS, immunocytochemistry method was used to examine the expression of CD133 in bone marrow cells of patients with leukemia and MDS. The results showed that the positive rate of CD133 in 41 acute leukemia patients was 51.2%. The expression of CD133 in AML patients (16/29, 55.2%) was significantly higher than that in control group (2/15, 13.3%). There was no significant difference in CD133 expression between CML and control group. The positive rate of CD133 in 9 patients with MDS was 55.56% (5/9). There was no significant difference between MDS and normal control. The expression of CD133 in all leukemia cells with CD34(+) was higher than that in leukemia cells with CD34(-), and there was significant difference in expression of CD133 between them (P < 0.05). The expression of CD133 had no relationship with the clinical prognostic factors such as sex, age, the percentage of leukemic cells in peripheral blood and in bone marrow, WBC counts, hemoglobin concentration, platelet counts and LDH level. It is concluded that the expression of CD133 in bone marrow cells of patients with AML is higher than that in control group. The expression of CD133 is significantly correlated with the expression of CD34. The high expression of CD133 may be an adverse prognostic factor in acute leukemia.
AC133 Antigen ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Antigens, CD34 ; immunology ; metabolism ; Bone Marrow Cells ; immunology ; metabolism ; Child ; Female ; Glycoproteins ; metabolism ; Humans ; Immunohistochemistry ; Leukemia, Myeloid, Acute ; immunology ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; Peptides ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; metabolism ; Prognosis ; Young Adult

OBJECTIVETo develop a convenient and effective method for the identification of Bungarus multicinctus.

METHODBased on the sequence of Cyt b gene fragment of B. multicinctus and its adulterants, a pair of highly specific primer (HJL- and HJH-) were designed for distinguishing B. ulticinctus from other species of snake. To establish specific PCR reaction condition, the primers were employed to amplify the DNA templates extracted from B. multicinctus and 6 other species of snake, under different annealing temperature. Using this method, B. multicinctus was identified from 18 samples bought from many drugstores.

RESULTA 230 bp DNA fragment was amplified from B. multicinctus in PCR with annealed temperature at 67 degrees C, whereas no DNA fragment was amplified from other snake samples under the same reaction condition, B. multicinctus could be clearly distinguished from others by PCR reaction with the highly specific primers. In the present study, 18 sample, bought from different drugstores, were also identified by the highly specific PCR with the primers. The results indicated that 14 samples were B. multicinctus and the other 4 were adulterant, which was consistent with the conclusion of authentication based on morphological.

CONCLUSIONThe primers designed in the present study were highly specific for B. multicinctus.

Animals ; Base Sequence ; Bungarus ; classification ; genetics ; Cytochromes b ; genetics ; DNA ; genetics ; DNA Primers ; Drug Contamination ; Materia Medica ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; Snakes ; classification ; genetics ; Species Specificity

This study was aimed to explore the expression of erythropoietin receptor (EPOR) on acute leukemia cells and its clinical significance. Bone marrow of 40 patients with acute leukemia (AL) and 24 patients with normal bone marrow as control group were collected. Samples came from outpatients and inpatients in our hospital. EPOR mRNA was detected by reverse transcription-PCR. The results showed that there was EPOR expression on AL cells, the expression rate was 57.5%, and the average expression level (Gray value) was 0.3549 ± 0.2800, but both were lower than that in control group (p < 0.05). There was no significant statistic difference of expression rate between acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL) (p > 0.05), and expression level of AML EPOR was higher than that of ALL (p < 0.05). It is concluded that there is EPOR expression on AL cells, while the expression rate and expression level are lower than those in control group (p < 0.05). There is no significant statistic difference of the expression rate between AML and ALL (p > 0.05), and the expression level of AML EPOR is higher than that of ALL (p < 0.05).
Case-Control Studies ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, Erythropoietin ; genetics ; metabolism

To investigate the effect of vitamin D in pathogenesis and clinical treatment of patients with immune thrombocytopenic (ITP), ELISA was used to detect the level of 25-hydroxylate vitamin D3[25(OH)D3] and 1,25-dihydroxyvitamin D3[1,25(OH)2D3] in peripheral blood from 45 ITP patients and 30 normal controls. Vitamin D receptor (VDR) mRNA expression was detected by RT-PCR. The results showed that the levels of 25(OH)D3 (10.6 ± 7.7 ng/ml) and 1,25(OH)2D3 (69.9 ± 29.0 pg/ml) in peripheral blood of the initial ITP patients were lower than those in peripheral blood of normal controls (13.7 ± 9.1 ng/ml, 87.3 ± 19.9 pg/ml) (P < 0.05). The expression of VDR gene obviously increased in peripheral blood of initial ITP patients (1.588 ± 0.127), as compared with that in peripheral blood of normal controls (1.055 ± 0.734) (P < 0.05). It is concluded that vitamin D level and its receptor expression may play an important role in ITP, and vitamin D and its similarities may be a new agent to treat patients with ITP.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Cells, Cultured ; Female ; Humans ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Receptors, Calcitriol ; blood ; Thrombocytopenia ; blood ; Vitamin D ; blood ; Young Adult

BACKGROUNDRadiation is a promising treatment for in stent restenosis and restenosis following percutaneous transluminal coronary angioplasty, which has troubled interventional cardiologists for a long time. It inhibits neointima hyperplasia, vascular remodeling, and increases the mean luminal diameter. The mechanism of intracoronary brachytherapy for restenosis is not well understood. Endogenous gaseous transmitters including nitric oxide and carbon monoxide are closely related to restenosis. Hydrogen sulfide, a new endogenous gaseous transmitter, is able to inhibit the proliferation of vascular smooth muscle cells and vascular remodeling. This study aimed to clarify the effect of radiation on cystathionine-gamma-lyase/hydrogen sulfide pathway in rat smooth muscle cells.

METHODSWe studied the effect of radiation on the cystathionine-gamma-lyase/hydrogen sulfide pathway. Rat vascular smooth muscle cells were radiated with (60)Co gamma at doses of 14 Gy and 25 Gy respectively. Then the mRNA level of cystathionine-gamma-lyase was studied by quantitative reverse-transcription competitive polymerase chain reaction. Hydrogen sulfide concentration in culture medium was determined by methylene blue spectrophotometry. Cystathionine-gamma-lyase activity in vascular smooth muscle cells was also studied.

RESULTS(60)Co gamma radiation at a dose of 1 Gy did not affect the cystathionine-gamma-lyase/hydrogen sulfide pathway significantly. However, (60)Co gamma radiation at doses of 14 Gy and 25 Gy decreased the hydrogen sulfide synthesis by 21.9% (P<0.05) and 26.8% (P<0.01) respectively. At the same time, they decreased the cystathionine-gamma-lyase activity by 15.1% (P<0.05) and 20.5% (P<0.01) respectively, and cystathionine-gamma-lyase mRNA expression by 29.3% (P<0.01) and 38.2% (P<0.01) respectively.

CONCLUSIONAppropriate (60)Co gamma radiation inhibits the H(2)S synthesis by inhibiting the gene expression of cystathionine-gamma-lyase and the cystathionine-gamma-lyase activity.

Animals ; Cells, Cultured ; Cobalt Radioisotopes ; Cystathionine gamma-Lyase ; genetics ; metabolism ; Enzyme Activation ; drug effects ; radiation effects ; Gamma Rays ; Hydrogen Sulfide ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; radiation effects ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; radiation effects

OBJECTIVETo establish a molecular method for the authentication of Pinellia pedatisecta and its adulterants.

METHODDNA sequences of some species from P. tenore, Typhonium and Arisaema were downloaded from GenBank, the sequences were aligned using DNAMAN. Allele-specific primers for P. pedatisecta and P. tenore were designed according to their SNPs in rpl 20 sequence. The designed primers were used to amplify 10 samples of P. pedatisecta, P. ternata and T. flagelliforme.

RESULTA 351 bp band was amplified from P. pedatisecta but not form P. ternata and T. flagelliforme by primer Pprpl149F and Pprpl484R. A 630 bp band was amplified from P. ternate and P. pedatisecta but not from T. flagelliforme by primer Ptrpl94F and Ptrpl699R.

CONCLUSIONAS-PCR has the advantages of highly specific and good reproducibility, by which P. pedatisecta can be identified from part of its adulterants quickly. It is a potential method to be used in the molecular identification of other materia medica.

Alleles ; China ; Consumer Product Safety ; DNA Primers ; genetics ; Pinellia ; genetics ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; Quality Control

OBJECTIVETo assess the response in THP-1 treated with Rv3671c protein in Mycobacterium tuberculosis (M.tuberculosis).

METHODSThe gene encoding Rv3671c protein of M.tuberculosis was cloned into pET-28a vector and then expressed in Escherichia coli. The Rv3671c was purified with Ni-NTA affinity and ion exchange chromatography. The detection of protein concentration was by Lowry method.THP-1 cell was stimulated with Rv3671c protein and cells were analyzed by Hochest staining under fluorescence microscopy to assay cell death (apoptosis and necrosis). TNF-α and IL-1β were detected by ELISA at each stimulating time.

RESULTSThe Rv3671c protein of M.tuberculosis was successfully expressed in Escherichia coli. The purity of recombinant Rv3671c protein was 95%, and the protein concentration was up to 0.4 mg/ml. The nucleus of THP-1 was isolated and necrosis-like under fluorescence when cells were stimulated by Rv3671c protein. The levels of TNF-α and IL-1β in supernatant were 19 000 and 16 500 pg/ml respectively, and were significantly higher than control cells with the levels of 2100 and 3800 pg/ml separately.

CONCLUSIONThe necrosis of THP-1 cells could be stimulated by Rv3671c protein of M.tuberculosis and it was probably associated with high cytokines TNF-α and IL-1β levels.

Bacterial Proteins ; pharmacology ; Cell Death ; Cell Line ; Humans ; Interleukin-1beta ; metabolism ; Macrophages ; cytology ; metabolism ; Mycobacterium tuberculosis ; genetics ; Tumor Necrosis Factor-alpha ; metabolism

This study was aimed to investigate the changes of the platelet particle membrane protein (GMP-140), platelet activating factor (PAF) and platelet parametes in the patients with hyperuricemia (HUA), ELISA was used to detect the levels of GMP-140 and PAF in 55 patients with HUA and 30 healthy individuals. Platelet parameters were measured with automatic blood cell analyzer, and the biochemical indexes were detected at the same time. The results showed that the levels of serum uric acid, triglycerides (TG) and low density lipoprotein cholesterol (LDL-C) in HUA patients were higher than that in the normal group (P < 0.01). Serum uric acid level of HUA group was higher in men than that in women. The levels of GMP-140 and PAF in HUA patients were much higher than that in the normal group (P < 0.01), the indexes of platelet distribution width (PDW) and platelet-large cell ratio (P-LCR) in HUA patients were higher than that in the normal group (P < 0.01), there was no statistically significant difference in platelet count, plateletcrit (PCT), mean platelet volume (MPV) between the two groups. There was positive correlation between serum uric acid and levels of GMP-140, PAF, P-LCR and PDW, respectively (r = 0.667, 0.879, 0.310, 0.460, P < 0.01 or P < 0.05). Multivariate stepwise regression analysis revealed that serum uric acid, creatinine, P-LCR, urea nitrogen contributed to GMP-140 level (adjusted R(2) = 0.822). Serum uric acid and LDL-C also contributed to PAF level (adjusted R(2) = 0.451). It is concluded that a close relationship exists between HUA and the change of platelet function, and HUA plays a certain role in cardiovascular disease thrombosis complications.
Adult ; Aged ; Blood Platelets ; metabolism ; Case-Control Studies ; Female ; Humans ; Hyperuricemia ; blood ; Male ; Middle Aged ; P-Selectin ; metabolism ; Platelet Activating Factor ; metabolism ; Platelet Count

To investigate the changes of plasma suPAR level in patients with multiple myeloma, ELISA was used to detect plasma suPAR level, and routine examination was performed for other clinical indexes in 26 multiple myeloma patients. The results showed that plasma suPAR level in patients was 4.31+/-1.67 ng/ml, which was obviously higher than that in control group (1.87+/-0.27 ng/ml) (p<0.01). Plasma suPAR level in IgM subtype patients was 6.18+/-3.61 ng/ml, which was highest among all the subtypes; the plasma suPAR levels in non-secretion subtype, IgG and IgA subtype were 4.43+/-1.55 ng/ml, 4.00+/-0.95 ng/ml and 3.50+/-1.60 ng/ml respectively. The plasma suPAR levels in all subtypes were higher than that in control group, but there was no differences between these subtypes. SuPAR level was correlated with the blood sedimentation rate, creatinine level and hemoglobin level, plasma cell ratio and M protein level. It is concluded that the change of plasma suPAR level in multiple myeloma contributes to predict the development and prognosis of the disease.
Aged ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; blood ; classification ; Prognosis ; Receptors, Urokinase Plasminogen Activator ; blood ; Solubility