Expression of NDV HN protein in rice and development of a semi-quantitative rapid method for detection of antibodies / 生物工程学报

The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log2, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.

Clinical effect of percutaneous intraluminal radiofrequency ablation combined with biliary stenting and biliary stenting alone in the treatment of malignant obstructive jaundice / 军事医学

Objective To compare the effect of percutaneous intraluminal radiofrequency ablation ( RFA ) combined with biliary stenting and that of percutaneous transhepatic puncture combined with biliary stenting .Methods A total of 56 patients with unresectable malignant obstructive jaundice were reviewed retrospectively .Among them, 25 patients had received percutaneous intraluminal RFA combined with biliary stenting ( RFA group) while another 31 patients had been simultaneously selected for the simple biliary stent implantation ( stent group ) .The changes of the serum total bilirubin ( TB) and direct bilirubin ( DB) before and after 7-14 days of treatment , surgical complications , stent median patency and the median survival were observed.Follow-up information was obtained through telephone reviews or check-up records. Results The technical success rate was 100%.No procedure-related peritonitis or perforation occurred .There were respectively 3 cases with cholangeitis in RFA group and 3 in stent group.All the cases was controlled by effective clinical treatment.There was obvious statistically significant difference after treatment in TB and DB in the two groups (P<0.01, P<0.01).TB and DB fell by (149.05 ±110.71) and (96.93 ±69.12)μmol/L after 7-14 days in RFA group vs (151.40 ±94.47) and (94.21 ±67.36)μmol/L in stent group.The changes of the two groups were of no statistical significances .The stent patenmedian time was 122 ( 9 -550 ) and 157 ( 16 -510 ) d, while the median survival was 125(9-550) and 163 ( 16 -520 ) d.The difference was of no statistical significance .Conclusion Percutaneous intraluminal RFA combined with biliary stenting and percutaneous transhepatic puncture combined with biliary stenting are both safe and feasible therapeutic options for unresectable malignant obstructive jaundice .There is no statistically significant diffference between the two groups in the recent and long-term curative effects .

Research about the effects of different preoperative fasting water time on body temperature in preoperative children / 中国实用护理杂志

Objective To investigate the effects of different preoperative fasting water time on body temperature in preoperative children. Methods Divided 350 children into A, B and C group randomly and apply 4 hours preoperative fasting water, 4 hours fasting water added 250～500 ml fluid infusion and 2 hours fasting water respectively. Observe the condition of body temperature, thirst, fidget, crying and vomiting in these 3 groups. Results The incidence rate of preoperative fever in group A was significant higher than that of in the other two groups. The incidence rate of thirst, fidget and crying were significant higher in group A and B than those of in group C, while there was no significant difference among three groups in the incidence rate of vomiting during the course of operation. Conclusion It is a safe method of 8 hours preoperative fasting food and 2 hours fasting water for children, which can effective reduce the incidence rate of preoperative fever and then guarantee the operation.