title: [The modified extracorporeal photochemotherapy promotes apoptosis of spleen lymphocytes].
abstract: OBJECTIVE
  To explore the efficacy of the modified extracorporeal photochemotherapy (ECP) in improving the apoptotic rate of lymphocytes in vitro.
  
  
  METHODS
  The spleens which were obtained from liver transplantation donor under aseptic condition were used as experimental materials. Splenic lymphocytes (SPs) suspensions were prepared by modified and traditional ECP method, respectively. And then the isolated SPs were treated by the irradiation of 8-methoxypsoralen (8-MOP) combined with ultraviolet A (UVA) named PUVA, 8-MOP and UVA, and compared with a blank group meanwhile. The treated SPs were cultured overnight in an incubator at 37 Degrees Celsius, in a humidified atmosphere of 50 mL/L CO2 for 6-8 hours. The morphological changes of cells were observed using an inverted microscope, the apoptotic rates of SPs were detected by flow cytometry, and the difference between groups was analyzed finally.
  
  
  RESULTS
  The apoptotic rate at early stage and the total apoptotic rate of SPs prepared by the modified ECP method were respectively (95.33±3.03)% and (97.10±2.12)% after treated by PUVA, (23.39±4.55)% and (36.32±6.63)% after treated by 8-MOP, and (66.98±3.60)% and (68.65±4.35)% by UVA. Compared with control group (12.82±1.86% and 13.4±2.65%), there were statistically significant differences (P<0.01). The apoptotic rate at early stage and the total apoptotic rate of SPs prepared by the traditional ECP method were respectively (79.73±4.21)% and (82.70±4.13)%, (61.42±2.28)% and (68.91±2.18)%, (19.30±1.78)% and (28.06±1.88)%, (10.84±0.98)% and (12.77±1.22)%, and the statistical comparisons between groups also had significant difference (P<0.01). In addition, there was a significant difference in the early and total apoptosis between the modified and traditional ECP (P<0.01), but no obvious variation in the end-stage apoptosis in the two groups (P>0.05).
  
  
  CONCLUSIONS
  The modified ECP method can promote apoptosis of SPs in vitro conveniently, safely and efficiently, especially in the early stage. This can lay a foundation for the further study on dendritic cell immunomodulation induced by ECP method.
authors.affiliation: Department of Hepatobiliary Surgery, General Hospital of PLA, Beijing 100853; Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Guosheng
authors.initials: G
authors.lastname: Du
authors.affiliation: Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Dehua
authors.initials: D
authors.lastname: Zheng
authors.affiliation: Department of Urology Surgery, Institute of Organ Transplantation, No. 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Zhijia
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Urology Surgery, Institute of Organ Transplantation, No. 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Yuxiang
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Department of Hepatobiliary Surgery, General Hospital of PLA, Beijing 100853, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Zhidong
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Song
authors.affiliation: Department of Hepatobiliary Surgery, 309th Hospital of PLA, Beijing 100091, China.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Fu
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000528:radiation effects*; D002478:Cells, Cultured; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects* / Q000528:radiation effects*; D008730:Methoxsalen / Q000494:pharmacology; D011701:PUVA Therapy / Q000379:methods; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000494:pharmacology; D015203:Reproducibility of Results; D013154:Spleen / Q000166:cytology; D013997:Time Factors; D014466:Ultraviolet Rays
chemical_list: D017319:Photosensitizing Agents; D008730:Methoxsalen