| Objective:Professor Huang Shi-Lin is a famous veteran teran doctor of TCM Who is an expert in treating hematological deseases with traditional Chinese medicine and integration of traditional and western medicine.He takes driving away evil and recovering healthy energy as the treatment principle for acute leukemia.And under this principle,he is the first to adopt autologous bone marrow transplantation(ABMT) purged with psoralen extracted from psoralea fruits plus long wave ultraviolet A(PUVA) to treat acute granulocytic leukemia(AGL).The objective of this study is to comprehensively summarize the clinical curative effect and Professor Hang Shi-Lin's invaluable clinical experience.Lastly,the study will explore the antileukemia mechanism of PUVA.Methods:From October 1991 to November 2005,28 patients with AGL underwent ABMT purged with PUVA were involved in the clinical study.The human leukemia cells HL-60,NB4 and K562 were taken as the object in the empirical study and affymetrix human U133 2.0 gene expression chips were used to detect the effects of PUVA on gene expression of leukemia cells.The changes of mitochondria membrane potential and expression of Fas and FasL protein after 24 hours of PUVA were detected with flow cytometry(FCM).The techniques about fluorescent quantitation RT-PCR and immunocytochemistry(ICC)were used to detect the expression of Fas and FasL mRNA after 4 hours of PUVA and Caspase8 and Caspase3 protein respectively.We used SPSS 11.5 software to statistically analyze the data.Results:1.The median of mononuclearcell before and after being purged with PUVA of 28 patients in bone marrow were 8.65×10~9 and 9.93×10~9 respectively (p=0.418＞0.05).The median percentage of CD34~+ mononuclearcell before and after being purged of 25 patients were 1.96%and 1.77%respectively(p=0.543＞0.05).2.27 of 28 patients achieved haematogenesis reconstruction,with a median of 13 days to reach an absolute meutrophil count(ANC)＞0.5×10~9/L and a median of 16 days to reach a platelet(PLT)count＞40×10~9/L.3.During the pre-treatment,28 patients all feeled indisposition in gastrointestinal tract and 11 of them vomited.During the restoration process of ABMT,3 patients were infected and had fever and hemorrhage occured in 2 other patients.4.8 of 28 patients relapsed on median of 7 months after ABMT and the total relapse ratio was 28.57%.The relapse probability was 22.22%for 1 year,and 29.63%for 2,3,5,7,10 years.5.With median follow-up of 52.5 months,the average survival time was 143.75±15.63 months(95%CI,.113.11,174.39)and the one year survival rate was 82.14%,75%for 2 years,71.43%for 3,5,7 and 10 years.The one year survival probability was 81.74%,73.22%for 2 years,71.43%for 3,5,7 and 10 years.6. The results of genechip showed that protein metabolism,fold and modification of protein structure and signal transduction pathway mediated by cell surface receptor related to cell apoptosis were mainly involved in the effect of PUVA on antileukaemia. Apoptosis genes and other relavent genes were activated.7.The results of FCM showed that mitochondria membrane potential and expression of FasL protein in HL-60,NB4 and K562 cells were obviously decreased and expression of Fas protein were obviously increased after 24 hours of PUVA.8.The expression of Fas and FasL mRNA were upregulated and downregulated respectively detected by fluorescent quantitation RT-PCR after 4 hours of PUVA.9.The results of ICC showed that PUVA promoted the expression of Caspase8 and Caspase3 protein.Conclusions:1.It is an innovation that PSO was used in ABMT for AGL based on its photosensitizing characteristics and antileukemia effect of PUVA.2.ABMT purged with PUVA in vitro for AGL does little harm to hematopoietic cells and can reduct replase and prolong desease free survival.3.PUVA can activate apoptosis and its relavent genes of leukemia cells 4.Fas/FasL system is one of the pathway about leukemia cells' apoptosis induced by PUVA,which can activate Caspase8 and Caspase 3.The changes of mitochondria membrane potential are involved in this process. |
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