Effect of angular pyranocoumarin isolated from peucedanum praeruptorum on the proliferation and apoptosis of U266 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effects of angular pyranocoumarin (±) -4'-O- acetyl-3'-Oangeloyl- cis- khellactone (APC) extracted from peucedanum praeruptoruon on the proliferation and apoptosis of U266 cells, and to explore its related mechanism.</p><p><b>METHODS</b>APC was extracted by petroleum ether technique, and its purity was tested by high performance liquid chromatography, and its chemical structure was identified by magnetic resonance spectroscopy. U266 cells were treated with APC in various concentrations (0, 10, 20, 30, 40 μg/ml）for different durations（24 and 48 h). The inhibitive effect of APC on cell growth was detected by CCK-8 method. After U266 cells were incubated with APC（0, 10, 20, 30, 40 μg/ml）for 24 h, the apoptosis of cells were observed by flow cytometry stained with Annexin Ⅴ/PI and Hochest33342; the expression levels of caspase-3, 8, ERK, p-ERK, AKT and p-AKT protein were assayed by Western blot; the expression of hTERT mRNA was measured by RT-PCR.</p><p><b>RESULTS</b>The purity of APC identified by magnetic resonance imaging was 98.8%. The proliferation of U266 cells was inhibited, and the apoptosis was induced in a time- and/or dose- dependent manner after treatment with APC. APC could upregulate the caspase- 8, 3 protein expression and downregulate the p- ERK, p-AKT protein expression along with the increase of APC dose. APC also could downregulate the hTERT mRNA expression.</p><p><b>CONCLUSION</b>Angular pyranocoumarin APC could inhibit the proliferation and induce the apoptosis of U266 cells. The probable mechanism might be achieved by upregulating caspase-8, 3 protein expression and downregulating p-ERK, P-AKT protein and the hTERT mRNA expression.</p>

Rituximab treatment for adults with steroid-resistant idiopathic thrombocytopenic purpura / 中华内科杂志

Objective To investigate the efficacy and safety as well as the effects of rituximab on B-lymphocytes and anti-platelet glycoprotein-specific antibodies,in patients with steroid-resistant idiopathic thrombocytopenic purpura(ITP).Methods Twelve steroid-resistant ITP patients,16 to 54 years old,received intravenous rituximab at the dose of 375 mg/m2 once-weekly for 4 weeks.Lab studies included CBC,serum concentrations of IgG,IgM and IgA.CD+3,CD+4,CD+8,CD+19,CD+20 cell numbers were assayed by flow cytometry and anti-platelet glycoprotein-specific antibodies(GP Ⅱ b/Ⅲ a,GP Ⅰ b/Ⅸ)were assayed by monoclonal antibody-specific immobilisation of platelet antigens prior to and following rituximab therapy.Results A complete response(platelet counts ≥100×109/L)was observed in 4 cases,a partial response (platelet counts between 50 and 100×109/L)in 3 cases,a minor response(platelet counts between 30 and 50×109/L)in 2 cases,and non response(platelet counts＜30×109/L)in 3 cases.Responses were sustained 0.5 to 12 months(median 5 months).After 4 weeks of rituximab therapy,anti-platelet glycoprotein-specific antibodies(GP Ⅱ b/Ⅲ a,GP Ⅰ b/Ⅸ)disappeared except one NR patient and CD+19/CD+20 cells were almost depleted in all patients(295.0±86.4)×106/L vs(4.1±2.2)×106/L(P＜0.01).As expected,the T cell counts,and the serum concentrations of IgG,IgM and IgA were not changed after therapy.No severe side effects were observed.Conclusion Rituximab may be an effective and safe treatment for adults with steroid-resistant ITP.

Experimental study of apoptosis induced by rhFasL gene transferred into primary cultured synoviocytes from patients with rheumtoid arthritis / 中华风湿病学杂志

Objective To observe apoptosis of primary cultured synoviocytes from patients with rheumtoid arthritis (RA) which induced by transfected recombinant plasmid carring Fas ligand (FasL) gene, aimed to detect the aim targets and develope gene therapy for RA by intra-articular injection. Methods FasL cDNA was introducted into the plasmid pcDNA3.1-neo by reverse transcriptase-polymerse chain reaction (RT-PCR) to set up the recombinated human FasL gene expression carrier, Then the pcDNA3.1-FasL was transducted into the primary cultured synovial cells by catholyte liposomes. After screened by G418, transgenic synovial cells were choosen. Finally through the microscope, TUNEL, Annexin Ⅴ-FITC and PI staining with flow cytometry, we observed the apoptosis state of each synovial cell. Results Prokaryotic and eukaryon carrier with FasL gene were established successfilly. The gene sequencing had completely coincident with the lierature. When eukaryon carrier with FasL (pcDNA3.1-FasL) was transducted into the index number livinging RA synovial cells after 15 h, most synovial cells presented distortion transformation, and the minority sheded from the bottom wall. The transgenic RA synovial cells after screened by G418, only a little amount survived about 1~2 per sight, and the cells grew slowly, the normal matched cells were cloned after 2~4 weeks. The FasL gene transgenic RA synovial cells presented positive findings by TUNEL staining. Early apoptosis cells by Annexin Ⅴ/PI staining, but the difference wasn′t significant. Conclusion The eukaryon carrier with FasL (pcDNA3.1-FasL) can be transferred into RA synovial cells. It can induce synovial cell apoptosis.