Objective:To investigate the clinical value of totally laparoscopic exclusion of splenic artery aneurysm combined with pericardial devascularization for portal hypertension com-plicated with splenic aneurysm.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 17 patients with portal hypertension complicated with splenic aneurysm who were admitted to 2 medical centers (15 cases in Shenzhen University General Hospital and 2 cases in Wuhan First Hospital) from January 2013 to May 2020 were collected. There were 7 males and 10 females, aged (59±14)years. All patients underwent totally laparoscopic exoclusion of splenic artery aneurysm combined with pericardial devascularization. Observation indicators : (1) surgical and postoperative conditions; (2) complications; (3) follow-up. Follow-up was conducted by out-patient examiantion and telephone interview to detect the effect of exclusion of arterial tumor, and blood re-flow, portal vein thrombosis and survival of patients 3 months after operation. The follow-up was up to December 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Results:(1) Surgical and postoperative conditions. All 17 patients successfully completed the operation, without perioperative death. The operation time, volume of intraoperative blood loss of 17 patients were (181±30)minutes, 187(range, 90?420)mL. The white blood cell count, red blood cell count, hemoglobin, serum albumin were (9±4)×10 9/L, (3.5±0.9)×10 12/L, (86±17)g/L, (36±7)g/L on the postoperative day 3. Time to postoperative abdominal drainage tube removal and duration of post-operative hospital stay were (7±4)days and (11±4)days. (2) Complications. All 17 patients had ascites after surgery, which were improved after oral treatment with diuretics. There was no complication such as intra-abdominal hemorrhage, gastrointestinal fistula, pleural effusion, infection, abscess formation, fever and vascular embolism. (3) Follow-up. All the 17 patients were followed up for 28.6(range, 7.0?84.0)months. During the follow-up, the splenic aneurysm cavity of all patients was completely isolated, no blood re-flow and no portal vein thrombosis was observed, and no patient died. Conclusion:Totally laparoscopic exclusion of splenic artery aneurysm combined with pericardial devascularization is safe and feasible in the treatment of portal hypertension complicated with splenic aneurysm.

Objective: To reduce the residual proteins and DNA of host cells in the preparation of H5N1 influenza A virus. Methods: Core 700 was firstly used to remove residual host cell proteins, and then Capto Q was used to remove host cell DNA. Several batches of H5N1 influenza A virus cultured in Madin-Darby canine kidney (MDCK) cells were purified using this method. The efficiency of purification was evaluated using many methods including quantitative real-time PCR, hemagglutination (HA) test and single radial immunodiffusion assay. Moreover, Benzonase nuclease was used for comparison. Results: Without the use of Benzonase nuclease, the overall removal rates of host cell DNA and residual proteins were 99.62% and 98.1%, and the HA antigen recovery rate was 66.96%. Conclusions This study established a purification strategy with good effect for cell-based influenza vaccines. It can efficiently remove host cell DNA and proteins and achieve a high HA recovery rate. The purification result is no worse than that of adding Benzonase nuclease, suggesting the potential of its application in actual vaccine production.

Objective: To investigate the best amount of TPCK trypsin in Madin Darby canine kidney (MDCK) cell suspension for the culture of H7N9 avian influenza virus. Methods: Different concentrations of TPCK trypsin were added during the periods of cell growth and virus production. Their effects on cell growth, viability, glucose and lactate metabolism, and hemagglutination titer were monitored every 12 h. Inter-batch differences were analyzed. The amount of trypsin added in the cell growth phase was 0, 1 μg/ml, 2 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml, 10 μg/ml and 15 μg/ml. The amount of trypsin added during the virus production period was 0, 0.5 μg/ml, 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml. When the hemagglutination titers were same, the adding amount was further optimized at different multiplicity of infection (MOI) of 0.001, 0.005, 0.025 and 0.05. Results: No significant linear effects of TPCK trypsin concentration on cell number, viability, and glucose and lactate metabolism were observed. No toxicity to cell growth was observed when TPCK trypsin concentration reached 15 μg/ml. After the inoculation of H7N9 avian influenza virus, the hemagglutination titers in the 1 μg/ml, 1.5 μg/ml, 2 μg/ml and 2.5 μg/ml TPCK trypsin groups reached the peaks at 48 h, which were 1∶2^6.5. At 60 h, the hemagglutination titers of the latter two groups decreased faster than those of the former two groups. When the MOI was 0.005, the hemagglutination titer of the 1.5 μg/ml group at 48 h was 2^6.5 higher than 2⁶ in the 1 μg/ml group under the same condition. There were differences between different batches of TPCK trypsin. Conclusions Adding 1 μg/ml and 1.5 μg/ml of trypsin could better promote the proliferation of H7N9 avian influenza virus, and 1.5 μg/ml of trypsin had a wider range of MOI applicability.