Objective:To investigate the effects of pulsed electric field (PEF) combined with gemcitabine (GEM) on the viability and stemness of HCCC-9810 cholangiocarcinoma stem cells.Methods:HCCC-9810 cholangiocarcinoma stem cells were established in serum-free, cytokine-rich medium and divided into four groups: the control group, GEM group, PEF group, and the pulsed electric field combined with gemcitabine (PEF+ GEM) group. Cell proliferation was detected using the Cell Counting Kit-8 (CCK8) assay. Cell viability and apoptosis rate were measured by flow cytometry. Cell invasion ability was assessed using the Transwell assay. The expression of stemness marker proteins CD133 and Octamer-binding transcription factor 4 (OCT4), as well as the expression of β-catenin, was detected by Western blotting.Results:Regarding cell viability, the GEM, PEF, and PEF+ GEM groups showed significantly lower cell viability and higher apoptosis rate than the control group at 24 h, 48 h, and 72 h (all P<0.05). At 48 h and 72 h, the PEF+ GEM group showed significantly lower cell viability (7.2%±0.3% and 5.9%±0.8%, respectively) than the GEM group (50.7%±0.6% and 31.0%±1.2%, respectively) and the PEF group (12.2%±0.2% and 12.8%±0.2%, respectively) (all P<0.05). Regarding stemness inhibition, the PEF+ GEM groups showed significantly lower expression levels of CD133 and OCT4 at 24 h, 48 h, and 72 h compared with the control group (all P<0.05). Notably, at 48 h, the PEF+ GEM group showed a significantly lower expression level of the OCT4 (0.61±0.02) than the GEM group (0.87±0.08) and the PEF group (1.00±0.10) ( P<0.01). Furthermore, at 24 h and 48 h, the GEM, PEF, and PEF+ GEM groups showed significantly lower expression levels of β-catenin compared with the control group (all P<0.05). Conclusion:Pulsed electric field combined with gemcitabine therapy demonstrated more effective anti-proliferation and cancer stemness inhibition effects on HCCC-9810 cholangiocarcinoma stem cells compared with either monotherapy.

Objective:To investigate the effects of pulsed electric field (PEF) combined with gemcitabine (GEM) on the viability and stemness of HCCC-9810 cholangiocarcinoma stem cells.Methods:HCCC-9810 cholangiocarcinoma stem cells were established in serum-free, cytokine-rich medium and divided into four groups: the control group, GEM group, PEF group, and the pulsed electric field combined with gemcitabine (PEF+ GEM) group. Cell proliferation was detected using the Cell Counting Kit-8 (CCK8) assay. Cell viability and apoptosis rate were measured by flow cytometry. Cell invasion ability was assessed using the Transwell assay. The expression of stemness marker proteins CD133 and Octamer-binding transcription factor 4 (OCT4), as well as the expression of β-catenin, was detected by Western blotting.Results:Regarding cell viability, the GEM, PEF, and PEF+ GEM groups showed significantly lower cell viability and higher apoptosis rate than the control group at 24 h, 48 h, and 72 h (all P<0.05). At 48 h and 72 h, the PEF+ GEM group showed significantly lower cell viability (7.2%±0.3% and 5.9%±0.8%, respectively) than the GEM group (50.7%±0.6% and 31.0%±1.2%, respectively) and the PEF group (12.2%±0.2% and 12.8%±0.2%, respectively) (all P<0.05). Regarding stemness inhibition, the PEF+ GEM groups showed significantly lower expression levels of CD133 and OCT4 at 24 h, 48 h, and 72 h compared with the control group (all P<0.05). Notably, at 48 h, the PEF+ GEM group showed a significantly lower expression level of the OCT4 (0.61±0.02) than the GEM group (0.87±0.08) and the PEF group (1.00±0.10) ( P<0.01). Furthermore, at 24 h and 48 h, the GEM, PEF, and PEF+ GEM groups showed significantly lower expression levels of β-catenin compared with the control group (all P<0.05). Conclusion:Pulsed electric field combined with gemcitabine therapy demonstrated more effective anti-proliferation and cancer stemness inhibition effects on HCCC-9810 cholangiocarcinoma stem cells compared with either monotherapy.

Objective To evaluate the dosimetry accuracy and clinical efficacy of 125I radioactive seed implantation using coplanar template (CPT) in the treatment of metastatic or recurrent chest wall tumor.Methods Thirty-one patients with metastasis or recurrence of chest wall tumor,who had been diagnosed with pathology between July 2005 and July 2015,were retrospectively studied.All patients underwent CPT-assisted 125I radioactive seed implantation.Brachytherapy radiation treatment planning system (BTPS) was used to make preoperative plans,and the prescribed dose (PD) was 110 Gy.CPT was used to assist CT guided 125I radioactive seeds implantation.Dose evaluation was performed immediately after implantation.The difference of dose parameters was compared between preoperation and postoperation,including Dg0,D100,V90,V100 and the numbers of seeds.Postoperative chest CT was conducted regularly to assess the treatment efficacy based on the response evaluation criteria in solid tumors (RECIST Version 1.1).The patients were followed up till July 2016.Results All patients went through implantation procedure successfully and there was no significant statistical difference between preoperative and postoperative dose parameters (P ＞ 0.05).The conformal index (CI) was 0.951 ± 0.13,external index(EI) was 6.5％ ±0.9％.Six months after implantation,CR,PR,SD and PD were 25.8％ (8/31),51.6％ (16/31),6.5％ (2/31) and 16.1％ (5/31),respectively.The effective rate was 77.4％,and local control rate was 83.9％ (26/31).Skin pigmentation occurred in 13 patients during the follow-up period,without any special treatment.Conclusions The auxiliary of CPT in the treatment of metastatic or recurrent chest wall tumor under the guiding of CT could achieve quality control,safety and effectiveness.

Objective To compare the planned radiation dose and the actual dose received after 125I radioactive seeds implantation supported by coplanar template (CPT) in lung cancer patients with mediastinal node metastases 4R,and to evaluate the clinical efficacy.Methods Totally 32 patients with lung cancer with mediastinal lymph node metastases 4R who had been diagnosed via cellular pathology studies were selected from January 2008 to December 2014.The mediastinal lymph node metastases were treated by CPT-assisted 125I radioactive seed implantation.Digital imaging and communications in medicine (DICOM) data were acquired by chest CT scan before implantation,brachytherapy radiation treatment planning system (BTPS) was introduced to carry out the plan,and the prescribed dose(PD) was 120 Gy.CPT was used to control the precision of needle penetration and implantation of radioactive seeds.Computer tomography (CT) was used to ensure the correct position of needles and radioactive seeds.Dose evaluation was performed immediately after implantation and was compared with planned dose using paired t-test.The 6-months postoperative chest CT was conducted to evaluate treatment efficacy according to response evaluation criteria in solid tumors(RECIST Version 1.1).Results All patients went through implantation procedure successfully.Dose evaluation after implantation was as followed.The average dose received (231.9 ±29.6)Gy,the dose received by 90％ of the target(D90) (150.8 ± 16.6) Gy,the dose received by 100％ of the target(D100) (100.4 ± 12.6)Gy,the volume of 100％PD covering the target(V100)(94.1 ± 2.6) ％,the volume of 200％ PD covering the target(V200) (33.0 ± 5.7) ％,the conformal index (CI) 0.75 ±0.06,the external index(EI) (22.7 ± 5.8)％,the average dose received by the superior vena cava (19.3 ± 7.2)Gy,and the average dose received by aorta (12.1 ± 5.1)Gy.Efficacy was followed for 6 months after implantation and the effective rate was 84.37％.There was no serious complications (such as radioactive lung injury,major vascular injury,bleeding,and et al.) occurred in follow-up period.Conclusions CPT assisted CT guided 125I radioactive seed imnplantation in treating mediastinal node metastases 4R can achieve preoperative BTPS,minimize major vascular or organ injury.It is an accurate,effective and safe treatment approach and may be of great value to standardize the procedure of radioactive seed implantation in mediastinal metastases.