Objective To produce anti-thrombomodulin antibodies.Methods Using genetic immunization: Eukaryotic expression plasmid pcDNA3.1/TM(LEO),encoding all the extracellular domain of human thrombomodulin and signal peptides but lacking the transmembrance and cytoplasmic domains was constructed, which recombinant thrombomodulin was secreted soluble product. The plasmid was isolated from large-scale bacterial cultures by treatment with alkali and SDS, purified by precipitation with polyethylene Glycol (PEG). Recombinant plasmid was injected into tibial muscle of BALB/c mice. The productions of TM and anti-TM have been detected. Results The positive of RT-PCR and expressed TM identified the function of the recombinant plasmid. The pcDNA3.1/TM(LEO) induced higher titer of anti-TM. The antibody titer peaked between the 5th and 7th injection with a titer of 1∶8 000 detected by cell-ELISA coated with EVC-304. Specificity has been identified by western blot and immunohistochemistry.Conclusion The production of antibody through genetic immunization was a feasible method due to the difficulties in obtaining and purification of natural thrombomodulin.

OBJECTIVETo investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C to IFN treatment.

METHODSThe HCV RNAs in PBMC were detected at the end of treatment, and 24 week and 1 year follow up after end treatment, in 16 patients who acquired complete response to IFN in 12 weeks of 24 weeks therapy.

RESULTS9 patients were HCV RNA positive in their PBMC at the end of treatment, the serum HCV RNA of 8 turned positive after 24 weeks and 1 year follow-up. In 7 patients with negative HCV RNA in PBMC, only two patients relapsed in serum HCV RNA after 1 year follow-up, and others remained viral response after 3.5 years.

CONCLUSIONHCV RNA in PBMC at the end of treatment was a predictor of the durable response to antiviral therapy in chronic hepatitis C.

Adult ; Female ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; RNA, Viral ; blood ; Recombinant Proteins

Objectives To investigate and evaluate the preventive and therapeutic effects of celecox ib,a selective cyclooxygenase-2(COX-2)inhibitor,on multiple colorectal adenorna and compare it with aspirin.Methods Ninty-six patients with colorectal multiple adenoma were randomly divided into A,B and C groups.Adenomas in all patients were removed with high-frequency eleetrocoagulation,electroexci- sion or argon plasma coagulation(APC)under colonoscopy.Then,group A were administered celecoxib 200 mg twice daily,group B aspirin 50 mg twice daily,group C served as control.Colonoscopy was per formed every 6 months in the first year,and every year in order to observe and evaluate the recurrence rate of adenoma and the side effects after the treatment.Results Twenty-seven patients in group A,26 pa- tients in group B and 27 patients in group C had completed the treatment.At the end of the treatment, on PP/ITT analysis,the cure rate of the eolorectal adenoma were 84.4%/100% ,78.1%/96.2% and 75.0%/88.9% in group A,B and C,respectively.During the first year of follow-up,there were 1 ,1 and 6 cases which were found recurrences of the adenomas in group A,B and C,respectively.The recurrence rates of coloreetal adenomas in group A(3.7%)and group B(4.0%)were significantly low er than that in group C(24.0%) (P＜0.05 and＜0.05,respectively).At the end of follow-up,the total recurrence rate of colorectal adenomas in group A(14.80%)and group B(19.2%)were significant- ly lower than that in group C(46.2%)(P＜0.05 and＜0.05).While the side-effective rate regroup A (3.3%)was significantly lower than that in group B(22.5%)(P＜0.05).Conclusions After re- section of the multiple colorectal adenomas,both the selective inhibitor of COX-2,celeeoxib and the non- selective inhibitor of COX-2,aspirin,may reduce its recurrence rate,but the former has a good tolerance and lower side-effects.

Objective:To observe the curative effect of different surgical arthroscopy on treatment for the discoid meniscus injury in knee joint.Methods: The clinical data of 78 patients with discoid meniscus injury who received treatment of arthroscopic minimally invasive surgical were retrospectively studied. 42 patients were complete discoid meniscus injury, and 36 patients were incomplete discoid meniscus injury in the 78 cases. The 42 patients with discoid meniscus injury of complete type were divided into observation 1 group (22 cases) and observation 2 group (20 cases), and the 36 patients with discoid meniscus injury of incomplete type were divided into control 1 group (19 cases) and control 2 group(17 cases). The patients of observation 1 group and control 1 group were treated with complete resection operation, and the patients of observation 2 group and control 2 group were treated with partial resection arthroplasty. The knee physiological function of all patients before and after treatment were assessed by using Lysholm knee function score and the subjective assessment table of International Knee Joint Documentation Committee (IKDC) for knee, and the curative effects of each group was compared.Results: After treatment, the Lysholm and IKDC scores of 4 groups significantly increased than that of before treatment(P<0.05), and the Lysholm and IKDC scores of observation 1 group was significantly higher than that of observation 2 group (t=9.193,t=9.065,P<0.05). But there was no statistically significant difference between the control 1 group and control 2 group about Lysholm and IKDC scores (t=0.874, t=0.891,P>0.05). The excellent rate of the observation 2 group was significantly lower than that of observation 1 group(x2=5.292,P<0.05), while the difference of the excellent rate between the control 1 group and the control 2 group was no statistically significant (x2=0.734,P>0.05).Conclusion: All of patients with discoid meniscus injury of knee joint receives different surgical treatment under arthroscopy can obtain good effect, but the curative effect of patients with discoid meniscus injury of complete type who receives completely excision is significantly better than that of who receives partial resection arthroplasty.

OBJECTIVE: To study DNA quantification and STR typing of samples pre-treated with pyramidon. METHODS: The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. RESULTS: In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. CONCLUSION Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology* ; Blood Stains ; DNA/isolation & purification* ; DNA Fingerprinting ; Forensic Medicine ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results ; Specimen Handling

The flavonoids were investigated from the whole plants of Lagopsis supina. The compounds were isolated and purified by various column chromatography, and their structures were identified by physiochemical properties and spectroscopic data. Two flavones were isolated from the CH2Cl2 layer of Lagopsis supina extract and identified as genkwanin (1) and 5-hydroxy-7,4'-dimethoxyflavone (2), respectively. Ten flavonoid glycosides were isolated from the water layer of Lagopsis supina and elucidated as kaempferol-3-O-6" (3-hydroxy-3-methylglutaryl) -β-D-glucoside (3), quercetin-3-O-6"-(3-hydroxy-3-methylglutaryl) -β-D-glucoside (4), quercetin-3-O-β-D-glucoside(5), kaempferol-3-Of3-D-glucoside ( 6), isorhamnetin-3-O-/-D-glycopyranoside (7), apigenin-7-O-6-D-glucoside (8), luteolin-7-O-β-D-glucoside (9), chrysoeriol-7-O-β-D-glucoside (10), rutin (11 ), and kaempferol-3-β-(6"-p-coumaroyl) -β-D-glucoside (tiliroside, 12). Compounds 3 and 4 were firstly isolated from Lamiaceae family, and compounds 1-12 were isolated from the plants of Lagopsis genus for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Lamiaceae ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.

OBJECTIVETo study the clinical features, diagnosis and therapy of hydroa vacciniforme-like cutaneous T cell lymphoma.

METHODSThe clinical presentations and the findings of laboratory examinations and skin biopsy of affected tissue in a child with hydroa vacciniforme-like cutaneous T cell lymphoma were retrospectively reviewed.

RESULTSThe child manifested as rash, fever and lymph node intumesce. Rash was pantomorphia, including edematous erythema, vesicles, crusts, necrosis and depressed scar, and it was mild in winter and severe in summer, mainly involving in the face and extremities. Epstein-Barre virus (EBV)-IgM was positive. Histopathological findings revealed focal lymphocyte invasion in subcutaneous panniculus adiposus, mainly surrounding the blood vessels. Immunohistochemistry showed CD3 (+), CD43 (+), CD20 (-), pax-5 (-), TIA (+), CD5 (+), CD8 (+), Granmye (+) and CD4 (-). The clinical symptoms were improved after glucocorticoid treatment in this child.

CONCLUSIONSHydroa vacciniforme-like cutaneous T cell lymphoma has special clinical manifestations. This disorder may be definitely diagnosed by skin biopsy of affected tissue and immunohistochemistry assay. Glucocorticoid treatment is effective. EBV infection may be related to the development of this disorder.

Child, Preschool ; Female ; Humans ; Hydroa Vacciniforme ; pathology ; Lymphoma, T-Cell, Cutaneous ; drug therapy ; immunology ; pathology ; Skin ; pathology ; Skin Neoplasms ; drug therapy ; immunology ; pathology

To easily obtain plasmin-alpha(2)-antiplasmin complex (PAP) with high purity, the purification procedure was improved by authors. After urokinase activated fresh human platelet-deficient plasma had been applied to Lysine-Sepharose 4B affinity chromatography column, elutions were sequentially performed by several eluents with different ingredients. The results showed that 3.0 mg PAP could harvested from 100 ml fresh plasma by this method and the whole procedure could be finished within several hours. In conclusion, this procedure is simple, rapid, economical and high-yield.
Blotting, Western ; Chromatography, Affinity ; Electrophoresis, Polyacrylamide Gel ; Fibrinolysin ; analysis ; isolation & purification ; Humans ; Sepharose ; analogs & derivatives ; alpha-2-Antiplasmin ; analysis ; isolation & purification

OBJECTIVEAllogeneic marrow transplantation is a curative therapy for thalassemia, but no more than 30% of patients have HLA-indentical sibling marrow donor. The selection of alternative donors of unrelative marrow and the study on the probability of treating thalassemia major with unrelated donor bone marrow transplantation are of importance.

METHODSNine children with thalassemia were included in the study, and their gene mutational type were homozygote of thalassemia and double heterozygote, respectively. All of them were finally diagnosed of thalassemia major, and treated with unrelated donor bone marrow transplantation. To high-resolution HLA typing, two patients were matched, five had one unmatched isoform and two had two unmatched isoforms. The erythrocyte blood type was not matched in six patients. The preparative regimen included busulfan (oral use, 16 mg/kg, divided for 4 days), cyclophosphamide (intravenous use, 200 mg/kg, divided for 4 days), antithymocyte immunoglobulin (intravenous use, 30 mg/kg, divided for 3 days), and fludarabine (intravenous use, 125 mg/m(2), divided for 3 days). Ciclosporin A and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis.

RESULTSAll patients had allergen reactions. One had hypotension. Five patients experienced I degrees approximately III degrees acute GVHD in the skin, while one had II degrees acute GVHD in liver. One patient had III degrees GVHD of intestines and gradually developed chronic GVHD in the skin, lungs and brain. One patient died of pulmonary hemorrhage. The duration when peripheral blood neutrophil count exceeded 0.5 x 10(9)/L was 12 - 26 days. The recovery time of WBC was as long as 23 - 110 days. Thrombocytes exceeded 50 x 10(9) within 61 approximately 142 days. The time when hemoglobin reached 100 g/L varied from 23 to 116 days. The last blood transfusion was on 13 - 62 days. Eight patients were fully grafted, while one was not grafted. During the 6 - 24 months of follow-up, seven patients' genotype of thalassemia major became normal. The erythrocyte blood type of five patients also changed into the same as that of donor. The hemoglobin was kept over 110 g/L without blood transfusion.

CONCLUSIONThe transplantation of unrelated donor bone marrow for thalassemia major was successful. Unrelated donor bone marrow transplantation could cure thalassemia major, which expanded the marrow donor source for the transplantation of thalassemia major.

ABO Blood-Group System ; Bone Marrow Transplantation ; adverse effects ; Child ; Child, Preschool ; Disease-Free Survival ; Female ; Follow-Up Studies ; Graft Rejection ; Graft Survival ; Histocompatibility Testing ; Humans ; Infant ; Male ; Transplantation Tolerance ; Transplantation, Homologous ; adverse effects ; Treatment Outcome ; beta-Thalassemia ; diagnosis ; therapy