Objective To investigate the activity of recombinant Vibrio vulnificus hemolysin (rVvhA) on the apoptosis of J774A.1 cells and the related mechanism. Methods The cytotoxic effect of rVvhA on the growth of J774A.1 cells was identified by MTT, celluar and mitochondrial morphology were observed by transmission electron microscopy, apoptosis or necrosis and mitochondrial membrane potential in J774A.1 cells were measured by flow cytometry, activities of caspase-3 ,-8,-9 were detected by spectrophotometry. Results The viability of J774A.1 cells exposed to rVvhA was inhibited, and it is dependent on dose. Celluar and mitochondrial uhrastructure both occurred to change obviously observed by transmission electron microscopy in J774A.1 treated by 2.0 HU/ml and 3.0 HU/ml rVvhA after 8 hours; and 3.0 HU/ml rVvhA group had a better cytotoxic effect on J774A.1 than that of 3.0 HU/ml rVvhA group. The percentage of apoptosis is (7.80±0.62)%, (12.33±0.12)%, respectively. Besides, the mitochondriai membrane potential also reduced, because the rate of fluorescence which is green increase 1.0% (normal) to 9.8% (2.0 HU/ml rVvhA) and 39.2% (3.0 HU/ml rVvhA). At the same time, the caspase-3, -9 activity increased gradually, but caspase-8 remained unchanging. In J774A.1 cells treated by 3.0 HU/ml rV-vhA + caspase-3 inhibitor(Ac-DEVD-FMK) or caspase-9 inhibitor(Ac-LEHD-FMK), The apoptosis of was reduced to(6.23±3.95)% ,(9.60±3.14)%, and the activity of caspase-3, -9 reduced, too. Conclusion The rVvhA has cytotoxic effect on J774A.1. Mitochondria-mediated apoptosis pathway which is dependent on caspase may be related to apoptosis induced by rVvhA in J774A.1.

Objective To investigate the changes of serum copper in mice after whole-body irradiation and analyze the feasibility of these changes as a biological dosimeter.Methods Serum copper in mice exposed to 60 Co γ-rays(0,1,2,3,5 Gy) was collected from the orbital of mice and detected with 5-Br-PADAP colorimetric method at 30 min and 7 d after radiation.One-way analysis of variance was used to analyze the difference of serum copper in each group and Dunnett-t test was used to compare the difference between control group and irradiated groups.Linear and quadratic linear fitting function was used to analyze the relationship between serum copper and radiation dose.The change of serum copper was detected at 30 min,1,3,5,7,10,13 and 16 d after radiation to observe the persistence of serum copper.The established relationships were used to estimate the dose in 8 mice irradiated by a blind dose.Results The amount of serum copper in irradiated mice were significantly (F =208.20,145.98,P ＜ 0.05)dependent on the radiation doses with dose responses of y =-0.091x + 0.936 and y =-0.011x2-0.032x + 0.962 (r =0.989,0.995) at 30 min and 7 d post-irradiation,respectively.The concentration of serum copper at 2.0 Gy decreased at 30 min post-irradiation,increased at 1-7 d,then kept at a stable level at 7-14 d even increased slightly after 14 d.With these dose response curves,after radiation with a blind dose of 2 Gy,the absorb doses of mice were assessed to be (1.83-2.25) Gy and (1.82-2.11) Gy at 30 min and 7 d in 95％ confidence interval,respectively.Conclusions The serum copper is a quick,simple,and sensitive biomarker for the early assessment of absorb dose of irradiated mice.

Objective The mediastinal mass usually posed higher risk of anesthesia and surgery due to its especial anatomical position,and this study aimed to analyze the perioperative anesthetic management of 3 patients undergoing resection of huge mediastinal mass,to identify the potential risks for this type of surgery and to summarize the corresponding perioperative anesthetic management protocol.Methods Three cases recently underwent resection of huge mediastinal mass in our hospital were reviewed.Their preoperative assessment and preparation,intraoperative anesthetic management,postoperative pain management and special interventions needed in the perioperative period were summarized and analyzed retrospectively.The enhanced recovery after surgery (ERAS) protocol was established for this type of surgery based on the analysis mentioned above,evidence reported at home and abroad and currently available ERAS protocols for other surgeries.Results Pleural malignant solitary fibrous tumor,thymus squamous cell carcinoma and malignant mixed germ cell tumor were the three mediastinal masses.The preoperative assessment focused on the impact of tumor on other vital organs within the mediastinum;anesthesiologists focused on massive hemorrhage and severe complications such as cardiac insufficiency and respiratory insufficiency;all the three patients were sent to the intensive care unit after surgery for further treatment,successfully recovered and were discharged;improving postoperative analgesia was helpful for recovery after resection of huge mediastinal mass.Conclusion Perioperative anesthetic management of patients undergoing resection of huge mediastinal mass is related to high risk,and establishing specific ERAS protocol is helpful in reducing complications and in promoting recovery after surgery based on the currently available evidence and characteristics of this type of surgery.