Adenosine Triphosphatases ; metabolism ; Animals ; Antioxidants ; metabolism ; Muscle, Skeletal ; drug effects ; enzymology ; Rats ; Sulfinic Acids ; pharmacology

Antigens ; analysis ; Cervical Intraepithelial Neoplasia ; metabolism ; Citric Acid ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; Edetic Acid ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; Intestinal Mucosa ; immunology ; Ki-67 Antigen ; analysis ; Microwaves ; Palatine Tonsil ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; analysis

Objective To explore the effects of thiamine and riboflavin on H2O2-induced DNA oxidative damage in human umbilical vein endothelial cell line ECV304.Methods ECV304 cells were incubated with 10,100,500,1000 mg/L of thiamine or 20,100,300,500 nmol/L of riboflavin for 24 h,and then oxidative damage of cells were induced by 25 mol/L H2O2 for 30 min.DNA damage was detected with single cell gel electrophoresis(SCGE)assay.ECV304 cells incubated without H2O2,thiamine and riboflavin were served as negative controls,and those incubated with H2O2 and without thiamine and riboflavin were served as positive controls.Results H2O2 induced DNA damage,and the indices of percent of DNA damage cells,percent of tail DNA,tail length and Olive tail moment were increased.The indices of cells pretreated with 10,100,500 mg/L of thiamine or 20,100,300 nmol/L riboflavin were significantly decreased(P0.05).Conclusion Proper supplementation of thiamine and riboflavin may decrease H2O2-induced DNA oxidative damage,while excess thiamine and riboflavin supplementation may be harmful to DNA and enhance the susceptibility to H2O2 potentially.

Antigens ; analysis ; Autoantigens ; analysis ; Breast Neoplasms ; metabolism ; Citrates ; Female ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; methods ; Iodide Peroxidase ; analysis ; Iron-Binding Proteins ; analysis ; Paraffin Embedding ; Receptors, Progesterone ; analysis ; Thyroid Gland ; immunology ; Tissue Fixation

Biopsy ; Humans ; Kidney ; pathology ; Staining and Labeling ; methods

OBJECTIVETo investigate the diagnosis and differential diagnosis of granulocytic sarcoma (GS).

METHODSThe morphological and immunological characteristics of 12 cases of GS were studied. FAB classification was made by peripheral blood, bone marrow picture and bone marrow biopsy assay.

RESULTSAll of the 12 cases presented with lymphadenopathy and soft tissue mass. Histologically, the tissue infiltration of GS was composed of blastic cells with round to oval nuclei showing an even, pale chromatin pattern. Some with cleaved or notched nuclei. There were prominent nucleoli and scant cytoplasm in the cells and mitosis was easily found. Immunohistochemically, CD(45) and lysozyme were positive in all of the cases, MPO in 11 (92%), CD(68) in 10 (83%), CD(34) in 5 (42%), and TdT in 2 cases (17%). CD(15) and Mac387 were mainly expressed in mature granulocytes. Examination of bone marrow sections and marrow aspirate smears showed that out of the 11 cases tested 8 were AML-M(2), 2 AML-M(1) and 1 AML-M(0). Only 1 case was nonleukemic, ie. solitary granulocytic sarcoma.

CONCLUSIONGranulocytic sarcomas are difficult to identify in routine paraffin-embedded tissue sections and usually misdiagnosed as non-Hodgkin's lymphomas. Immunohistochemistry study with a panel of antibodies in combination with bone marrow and peripheral blood examination are helpful in identification of granulocytic sarcoma.

Adolescent ; Adult ; Aged ; Antigens, CD34 ; analysis ; Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Leukocyte Common Antigens ; analysis ; Male ; Middle Aged ; Sarcoma, Myeloid ; diagnosis ; metabolism ; pathology

To study the effect of interleukin-15 (IL-15) on the proliferation, differentiation and apoptosis of MDS CD34(+) cells, CD34(+) cells of high enrichment were separated by MACS system, and cultured in liquid media with different concentration of IL-15 in treated group and without IL-15 in the control group. Apoptosis of hematopoietic precursors was assayed by propidium iodine staining and cell by FCM, and the other MDS CD34(+) cells were stained by cytochemical staining after culture. The results showed that after culture with IL-15 the proliferation and differentiation of MDS CD34(+) cells were obviously promoted. It was found the every lineage of mature cells developed, the expressions of cell surface antigens CD71, CD33 and CD19 all increased in the MDS CD34(+) cell treated with IL-15. It is suggested that IL-15 stimulates the proliferation and differentiation of MDS CD34(+) cells, and partly shows anti-apoptosis effects which may be applicable to the therapy MDS.
Antigens, CD ; immunology ; Antigens, CD19 ; immunology ; Antigens, CD34 ; immunology ; Antigens, Differentiation, Myelomonocytic ; immunology ; Apoptosis ; drug effects ; Bone Marrow Cells ; drug effects ; immunology ; pathology ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Humans ; Interleukin-15 ; pharmacology ; Microscopy, Fluorescence ; Myelodysplastic Syndromes ; blood ; immunology ; pathology ; Receptors, Transferrin ; immunology ; Sialic Acid Binding Ig-like Lectin 3

OBJECTIVETo compare the immediate effect of moxibustion on Yanglingquan (GB 34) on body surface thermograph between the patients with myofascial pain syndrome (MPS) and healthy people.

METHODSThirty-five cases of MPS were selected in observation group and 15 cases of healthy people were in control group. ATIR-M301 medical infrared imaging device was used to observe the changes in body surface thermograph before and 15 min after moxibustion on Yanglingquan (GB 34) The qualitative analysis of picture pattern and the quantitative analysis of temperature were integrated to compare the differences between two groups.

RESULTSConcerning to the infrared thermograph expression on the back of MPS cases in observation group, the abnormal thermal zone presented in 88.6% of the cases, mostly distributed in the upper and secondly in the lower back. The spinal thermal line was broken or disappeared ine about 50% of cases. The location of pain complained by 71.4% of cases was coincident with the zone of infrared thermograph expression. There was statistical significant difference in the temperatures difference of the two sides in the upper abnormal thermal zone on the back in observation group as compared with control group (P < 0.001). Body surface thermograph on the back were impacted by moxibustion on Yanglingquan (GB 34) for either MPS patients or healthy people, especially in upper back and rhomboid fossa. Moxibustion on Yanglingquan (GB 34) increased much more apparently the temperature on the upper back in MPS patients as compared with healthy people (P < 0.05).

CONCLUSIONMoxibustion on Yanglingquan (GB34) brings the immediate improvements in the back thermograph for MPS patients.

Acupuncture Points ; Adult ; Aged ; Body Temperature ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Myofascial Pain Syndromes ; physiopathology ; therapy ; Thermography ; Young Adult

S1 nuclease (from Aspergillus oryzae) is a specific enzyme to degrade single stranded DNA or RNA molecules. It has been reported to be able to convert superhelical circular DNA molecules into open circle or linear forms under certain conditions, but this function has not been well explored. In order to use the action of S1 nuclease to linearize circular DNA and develop a novel way of cloning microcircular DNAs, the pUC19 was used to investigate the relationship between the linearization efficiency of S1 nuclease and the amount of enzyme used. By this way the optimal conditions for linearization of circular DNAs by S1 nuclease would be determined. 0.3u to 17u S1 nuclease per 100ng pUC19 DNA was added into a 25 microL system, respectively, to perform the reaction. The effectiveness of enzyme digestion was realized by electrophoresis in a 1.2% agarose gel. The results showed that along with the increase in enzyme amount from 0.3u to 17u a gradual decrease in the superhelical form, a gradual increase in the linear form and then in the circular form was obvious. The conversion from superhelical form to linear and circular form was directly related to the enzyme amount used. A higher proportion of linear DNA molecules was achieved by using 5 to 17u S1 nuclease per 100ng DNA. Besides, electrophoretic mobility of the S1 nuclease-linearized pUC19 was the same as that of the linear form produced by restriction enzyme digestion. According to the result of phiX174 digested by S1 nuclease it has been proposed that the enzyme cleaves first randomly on one site of one strand, thus converting the superhelical molecules into open circle form, and then on the same site of the complementary strand to produce the linear form. Therefore, the S1 nuclease-linearized DNA molecules are intact in the sense of their length and can be used for cloning. The plasmid-like DNA pC3 from cucumber mitochondria is a double stranded circular DNA molecule with about 550bp and the smallest known plasmid-like DNA in eukaryotic mitochondria. Many attempts have been made to linearize the molecule by using restriction enzymes but failed. Therefore, S1 nuclease was used to linearize pC3 based on the results obtained with pUC19. The linearized pC3 DNA molecules formed a very sharp band in a 2.5% agarose gel after electrophoresis. They were then recovered from the gel, added an "A" tail and ligated with T-vector. After transformation into E. coli JM109 cells, the positive clones were, screened by the blue-white selection. The insert was then cut using restriction enzymes EcoRI and Pst I. The result of electrophoresis shows that the electrophoretic mobility of the insert is just the same as that predicted. A 32 P-labled probe was synthesized using pC3 as the template and Southern blot analysis was carried out. The result shows that the inserted DNA is hybridized to the probe, which indicates that the cloned DNA fragment is from pC3. The sequence information of the insert shows that the plasmid-like DNA pC3 was 537bp in length. The nucleotide sequence was deposited in the GenBank (the accession number is AF522195).
Blotting, Southern ; Cloning, Molecular ; methods ; DNA, Circular ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Single-Strand Specific DNA and RNA Endonucleases ; genetics ; metabolism

OBJECTIVETo analyze treatment and prognostic factors of 74 patients with dermatofibro-sarcoma protuberans (DFSP).

METHODSFrom August 1990 to November 1999, 74 patients with DFSP confirmed pathologically were treated. There were 52 males and 22 females with a median age of 37 years (range 4 to 80 years) on diagnosis. Seventeen patients were treated by extensive excision and 2 by limited excision. Fifty-two patients had surgical resection alone (S), and 22 postoperative radiotherapy (S + R) of 50-70 Gy. The multivariate parameters were analyzed using Cox model. Kaplan-Meier and Log-Rank test were used to evaluate the results of the recurrence-free survival.

RESULTSThe rate of recurrence was 28.4% for all patients. The 5-year recurrence-free survival rate (RFSR) was 66.6% and the 10-year RFSR was 52.5%. The 5-year and 10-year in the S group were 58.4% and 41.2%, compared with 90.0% and 83.3% in the S + R group (P < 0.05). The 5-year and 10-year RFSR in the pathologically positive margin group were 57.5% and 41.4% respectively, compared with the 75.0% and 56.6% in the pathologically negative group (P < 0.05). Multivariate analysis suggested radiotherapy and negative pathological margins were favorable prognostic factors.

CONCLUSIONPost-operation radiotherapy and pathological margin are the independent prognostic factors.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Combined Modality Therapy ; Dermatofibrosarcoma ; mortality ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Postoperative Care ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Skin Neoplasms ; mortality ; radiotherapy ; surgery