The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

The purpose of this study is to investigate the effect of TPCK trypsin on the proliferation pattern of porcine epidemic diarrhea virus in Vero cells.The TPCK trypsin and conventional tryp-sin were added for virus proliferation,and RT-qPCR technology was used to analyze the changes in virus adsorption and invasion in Vero cells.The replication ability of porcine epidemic diarrhea vi-rus in Vero cells was explored through growth curve drawing,IFA identification,and cell activity detection.The results showed that the optimal concentrations of TPCK trypsin and conventional trypsin were 1 mg/L and 6 mg/L,respectively.The virus showed a decreasing trend with the pro-longation of TPCK trypsin and conventional trypsin pretreatment time.Adding different pancreatic enzymes during the virus proliferation process did not promote the virus invasion in Vero cells.Af-ter 4 h of invasion,the virus particles of each group gradually increased.By plotting the growth curve,it was found that the virus content of the TPCK trypsin group reached its highest level at 24 h(lgTCID50=(6.30±0.14)/0.1 mL),followed by a decreasing trend at 36 h,and the fluorescence intensity produced at 24 h was higher than that of conventional trypsin.In summary,TPCK trypsin has a better promoting effect on the proliferation of porcine epidemic diarrhea virus in Vero cells.It provided theoretical basis for further research on the mechanism of TPCK trypsin affecting porcine epidemic diarrhea virus proliferation,and also provided data support for the isola-tion and purification of porcine epidemic diarrhea virus epidemic strains.

Objective:To investigate the current status of positive feelings of caregivers of children with inflammatory bowel disease (IBD) and analyze the influencing factors.Methods:This study was a cross-sectional study. Using the convenient sampling method, a total of 288 caregivers of children with inflammatory bowel disease who were diagnosed and treated in Department of Gastroenterology of 2 children's hospitals in Henan Province were selected from September to December 2019. The general information questionnaire, Positive Aspects of Caregiving (PAC) and Caregiver Burden Inventory (CBI) were used for investigation. A total of 288 questionnaires were issued and 262 valid questionnaires were returned. The effective recovery rate was 90.97%.Results:Scores of PAC and CBI of caregivers of 262 children with inflammatory bowel disease were respectively (26.08±3.52) and (10.35±2.60) . Multivariate linear regression analysis showed that the payment method of medical expenses, education background of caregivers, religious beliefs, emotional burden and time-dependent burden were influencing factors of positive feelings of caregivers of children with inflammatory bowel disease ( P<0.05) . Conclusions:Clinical nursing staff should pay attention to the current situation of positive feelings of caregivers of children with inflammatory bowel disease and take targeted intervention measures to effectively improve the level of positive feelings of caregivers.

Objective: To establish a method of HPLC for determination of paeoniflorin in Yifu Capsules. Methods: HPLC analysis was carried out on ALLIMA C 18 column with acetonitrile water(15:85) as a mobile phase and detection wavelength at 230nm. Results: The linear range was 0.0798～0.7980?g (?=0.9998,n=6), and the average recovery was 99.00%(RSD=2.17%,n=5). Conclusion: The method is simple, accurate,highly sensitive and reproducible. It can be used for determination of paeoniflorin in Yifu Capsules.