Experimental study on topical treatment of diabetic skin ulcers with yi medicine "yi bu a jie" extract / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the role and mechanism of the Yi medicine, Yi Bu A Jie () extract, in topical treatment of diabetic skin ulcers, with a view to finding a breakthrough natural drug for the prevention and treatment of diabetic skin ulcers.</p><p><b>METHODS</b>A model of diabetic skin ulcers in Kunming mice was developed. Yi Bu A Jie was extracted in a Soxhlet extractor. Two different concentrations of the extract (0.005 mg/mL and 0.01 mg/mL) were applied to the wound of diabetic skin ulcers once every 3 days, and local skin appearance and histopathological changes were observed.</p><p><b>RESULTS</b>The shortest healing time was 25.25±2.06 day with a low concentration (P=0.0037 compared with the high concentration group, 33.14±2.21 day; P=0.0082 compared with control group, 28.21±2.14 days). The longest healing time was in the high concentration group (P=0.0025 compared with the control group). In both groups, a large number of inflammatory neutrophil cells were exuded during the experimental period. In the low concentration group, capillary-rich granulation tissue and actively growing fibroblasts appeared in the wound, while there was much necrotic tissue in the high concentration group.</p><p><b>CONCLUSION</b>Yi Bu A Jie extract has an inhibitory effect on diabetic skin ulcers in mice, and the low concentration is more suitable.</p>

Relationship of TGF-β and IL-4R gene polymorphisms with risk of classical Hodgkin lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to analyze the relationship between single nucleotide polymorphisms of transforming growth factor-β1 G-800A and C-509T, interleukin-4 receptor V75I and susceptibility of CHL in adults.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to analyze the expressed alleles of the selected SNP loca. The relationship between genomic polymorphisms of TGF-β1 and IL-4R and susceptibility of CHL were coupled with clinical data.</p><p><b>RESULTS</b>TGF-β1G-800A and TGF-β1C-509T had obvious linkage equilibrium (D' = 0.879, r(2) = 0.83, P = 0.020). GT haplotype distribution frequencies in mixed cellularity Hodgkin lymphoma cases and control group were of 53.1% and 34.2%, respectively, with statistically significant (OR = 2.35, P = 0.000); distribution frequencies of mutant gene T/T in disease and control groups were of 38.8% and 15.3%, respectively, also with statistically significant (OR = 3.654, P = 0.000); frequencies of nodular sclerosis CHL patients with IL-4R V75I mutant gene A/A in disease and control groups were of 19.2% and 41.75%, respectively, also with statistically significant (OR = 3.156, P = 0.000).</p><p><b>CONCLUSION</b>Single nucleotide polymorphisms of TGF-β1 G-800A, C-509T and IL-4R V75I has a significant correlation with Chinese susceptibility to classical Hodgkin lymphoma.</p>

Study of a humanized anti-platelet GP VI single chain Fv phage antibody / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the anti-platelet GPVI single chain Fv phage antibody which can inhibit the aggregation function of platelet by using phage antibody library technology.</p><p><b>METHODS</b>ITP patients with anti-platelet GPVI autoantibody that could inhibit the aggregation function of platelet were screened by MAIPA assay and platelet aggregation test. The gene fragments of heavy chain and light chain variable region (VH and VL) of immunoglobulin were amplified by RT-PCR from peripheral blood lymphocytes mRNA of the screened patients. The VH and and VL fragments were linked through a DNA linker encoding the peptide (Gly4Ser)3 to construct single chain Fv (ScFv) gene. The ScFv gene was digested with SfiI/NotI restriction enzymes and cloned into the pHEN2 phage display vector, then electrically transformed to E. coli TG1. The TG1 containing ScFv-pHEN2 was rescued by helper phage M13K07 to produce ScFv phage antibody. The anti-platelet GPVI phage ScFv antibody was enriched and purified. The effect of the phage antibody on platelet aggregation function was studied.</p><p><b>RESULTS</b>Of 806 chronic ITP patients, 11 (1.36%) were positive for anti-platelet GPVI autoantibody and 2 (0.24%) patients'plasma significantly inhibited the collagen induced platelet aggregation. The length of VH and VL fragments was about 380 to 400 bp, and were successfully formed ScFv fragments of about 800 bp by DNA linker. After cloning ScFv to phagemid vector pHEN2 and transforming ScFv-pHEN2 to TG1, 4.1x10(7) clones were obtained. After M13K07 rescue, 2.62x10(10) cfu/ml ScFv phage antibodies were produced. The purified anti-platelet GPVI ScFv phage antibody inhibited the collagen induced platelet aggregation.</p><p><b>CONCLUSION</b>Anti-platelet GPVI ScFv phage antibody produced by phage antibody library technology can inhibit the aggregation function of platelet.</p>

Significance of CD4+ CD25+ CD127(low) regulatory T cells and notch1 pathway in the pathogenesis of aplastic anemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To quantify the CD4+ CD25+ CD127(low) regulatory T cell (Treg), the expression levels of forkhead/winged helix transcription factor FOXP3 and Notch1 mRNA in aplastic anemia (AA) patients before and after treatment, and explore the significance of Treg in pathogenesis of AA.</p><p><b>METHOD</b>CD4+ CD25+ and CD4+ CD25+ CD127(low) T cells in peripheral blood were examined with FACS in 29 AA patients at active phase, 14 at recovery phase, 11 at unrecovery phase, and 15 normal controls. The levels of FOXP3 mRNA and Notch1 mRNA expression were detected with RT-PCR, and the correlations between Treg, FOXP3 mRNA and Notchl mRNA were analyzed.</p><p><b>RESULTS</b>The percentages of peripheral activated CD4+ CD25+ T cells in AA patients at active phase (4.3 +/- 0.7)% and unrecovery phase (4.2 +/- 0.6)% were significantly higher than those in normal controls (2.4 +/- 0.8)% (P < 0.05). The proportion of these cells in AA patients at recovery phase was reduced to (2.6 +/- 0.7)% (P < 0.05), being no difference from that in control group. The number of CD4+ CD25+ CD127(low) T cells in AA patients at active phase (2.4 +/- 1.2)% and unrecovery phase (2.5 +/- 1.1)% was decreased significantly compared with those in normal controls (7.1 +/- 2.7)% (P < 0.01) and in AA patients at recovery phase (5.3 +/- 1.0)% (P < 0.01), there was no difference between the latter two groups. In active phase AA patients, the levels of FOXP3 mRNA and Notchl mRNA (0.260 +/- 0.011 and 0.018 +/- 0.005, respectively) were lower than that in control group (1.307 +/- 0.011 and 0.308 +/- 0.028, respectively) (P < 0.01 and P < 0.01). After treatment, the levels significantly increased to 1.287 +/- 0.012 and 0.281 +/- 0.013 (P < 0.01 and P < 0.01), but there was no difference with that of normal controls (P > 0.05). CD4+ CD25+ CD2(low) T cells and FOXP3 were positively related with Notchl (P < 0.01) in AA patients.</p><p><b>CONCLUSION</b>The decreased number and suppressive activity of CD4 CD25+ CD127(low) Treg cells in the peripheral blood of AA patients cause over-activation of autoreactive T cells and suppression of haematopoiesis. One of the mechanisms maybe the reduced expression of Notch1 in the target cells.</p>

Study on the direct MAIPA technique in the differential diagnosis of immune and non-immune thrombocytopenia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical usefulness of direct monoclonal antibody immobilization of platelet antigen (MAIPA) technique in the differential diagnosis of immune and non-immune thrombocytopenia.</p><p><b>METHODS</b>Platelet-bound autoantibodies in thrombocytopenic patients (immune and non-immune) were measured by direct MAIPA. Monoclonal antibodies against GP II b/III a, GPIb and GP I a/II a were used.</p><p><b>RESULTS</b>The positive rates of platelet-bound GP-specific autoantibodies between immune (76.4%) and non-immune thrombocytopenia (3.6%) were significantly different (P < 0.05). The direct MAIPA had a sensitivity of 76.4%, a specificity of 96.4%, and a positive predictive value of 97.1% for the diagnosis of immune thrombocytopenia. There was a significant inverse correlation between platelet-bound GP II b/III a specific autoantibody levels and platelet counts (r = -0.338, P < 0.05).</p><p><b>CONCLUSION</b>The direct MAIPA technique can be used to differentiate immune from non-immune thrombocytopenias.</p>

Study on the T lymphocytes early activation and soluble tumor necrosis factor receptor in patients with aplastic anemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of T cell early activation marker (CD(69)) on peripheral CD(4)(+) and CD(8)(+) lymphocytes and serum levels of soluble tumor necrosis factor receptor 1 and 2 (sTNF-R1 and sTNF-R2) in serum and bone marrow in patients with aplastic anemia (AA) and their pathophysiological significance.</p><p><b>METHODS</b>In vitro activation of T lymphocytes was carried out by whole blood cell culture containing PHA (20 micro g/ml). The CD(69) expressions on CD(4)(+) and CD(8)(+) lymphocytes at 0 h and 4 h after PHA exposure were analyzed by two-color flow cytometry. The levels of sTNF-R1 and sTNF-R2 in serum and bone marrow were measured by ELISA.</p><p><b>RESULTS</b>The CD(69) expression rates of CD(4)(+) and of CD(8)(+) cells in SAA patients were (8.96 +/- 7.23)% and (10.67 +/- 7.58)%, respectively, and that of CD(8)(+) cells in CAA patients was (7.36 +/- 5.49)% before PHA stimulation. The CD(69) expression rates of CD(4)(+) and of CD(8)(+) cells in SAA patients were (71.73 +/- 11.91)% and (61.74 +/- 13.44)% and in CAA (59.35 +/- 10.15)% and (48.78 +/- 8.25)% respectively, and were significantly elevated after PHA stimulation. CD(69) expression on CD(4)(+) cells was much higher than that on CD(8)(+) cells after stimulation. The levels of the two sTNF-R (sTNF-R1 and sTNF-R2) in peripheral blood and bone marrow of SAA patients were elevated and in the bone marrow of CAA patients were also increased. The serum levels of sTNF-R2 were positively related to the CD(69) expression rates of CD(8)(+) cells before PHA stimulation.</p><p><b>CONCLUSION</b>Increased early activation and activated potentials of T lymphocytes, along with abnormally elevated immunologically active molecules might play a major role in the pathogenesis of AA.</p>