Objective To evaluate the immunogenicity of the recombinant human papillomavirus type 68b (HPV68b) virus-like particles(VLPs)in a mouse model.Methods The L1 protein of HPV type 68b was successful expressed in the Hansenula polymorpha strain (NVSI-68b-1).Processes including purifi-cation and reconstitution were performed to achieve pure HPV 68b VLPs.The purity, morphology and immu-nogenicity of the purified HPV 68 b VLPs were further analyzed .The BALB/c mice were immunized with HPV68b VLPs formulated on aluminum adjuvant .Pseudovirus-neutralizing antibody ( PsV NAb) assay was performed to detect the neutralizing antibodies in serum samples .Results The HPV 68 b L1 VLPs were ob-tained as indicated by the results of SDS-PAGE, Western blot assay , HPLC, electron microscopy and dy-namic light scattering with a high purity of 95%.Transmission electron microscopy and dynamic light scat-tering analysis revealed that the HPV68b L1 VLPs resembled the native virus with an average particle diame-ter of 50 nm.High levels of HPV68b-neutralizing antibodies were detected in serum samples from the mice immunized with HPV68b L1 VLPs.Moreover, a cross-protective efficacy of HPV68b L1 VLPs for HPV68a was observed .Conclusion This study suggested that the recombinant HPV 68 b VLPs expressed in a Han-senula polymorpha strain might be used as a potential candidate for the development of HPV vaccine .

Objective:To evaluate the diagnostic value of 99Tc m-pyrophosphate (PYP) in transthyretin cardiac amyloidosis. Methods:From December 2018 to July 2019, 17 patients (9 males, 8 females, age: (53.4±13.0) years) with suspected cardiac amyloidosis underwent 99Tc m-PYP imaging in Peking Union Medical College Hospital were prospectively included. Visual score and semi-quantitative values (heart to contralateral ratio, H/CL) of 99Tc m-PYP uptake were used to diagnose transthyretin amyloidosis (ATTR). Biopsies and genetic measurements were also developed to evaluate the diagnostic value of the imaging. Results:Five of the 17 patients were diagnosed as ATTR with a visual score of 2-3, H/CL≥1.5, and confirmed with the biopsy or gene test. Four patients were diagnosed as ATTR with positive genetic results but no cardiac symptoms, and their visual scores were between 0 and 1 with H/CL<1.5. Considering the young age of the patients, amyloid deposition might have not yet caused visceral damage. Visual score of other 8 patients with negative 99Tc m-PYP imaging were also between 0 and 1 with H/CL<1.5, 2 of 8 were confirmed with light chain amyloidosis (AL) by biopsy, 3 were clinically diagnosed as AL and 3 were ATTR excluded. The accuracy of 99Tc m-PYP imaging for diagnosing ATTR was 11/11. Conclusion:99Tc m-PYP imaging is helpful for non-invasive diagnosis of transthyretin cardiac amyloidosis.

OBJECTIVE：To prov ide reference for the improvement of quality standards of Shiwei yipi granules. METHODS ： According to the general rules of 2015 edition of Chinese Pharmacopeia （part Ⅳ），microscopic identification was used to identify Massa Medicata Fermentata and Galli Gigerii Endothelium Corneum ；TLC method was used to qualitatively identify Crataegi Fructus and Semen Raphani ；the content of sinapine thiocyanate in Semen Raphani was determined by HPLC. RESULTS ：The microscopic characteristics were obvious for Massa Medicata Fermentata （palisade cells of testa and stone cells of testa ）and Galli Gigerii Endothelium Corneum （irregular fragments ）. The same fluorescent spots of Crataegi Fructus and Semen Raphani were displayed at the same position of ursolic acid ，sinapine thiocyanate control and Semen Raphani reference substance. The linear range of sinapine thiocyanate was 23.27-9 574.42 ng （r＝1.000 0）. The LOD and LOQ were of 0.50 μ g/mL and 1.68 μ g/mL， respectively. RSDs of precision ，repeatability，intermediate precision and stability tests （24 h）were all less than 1.5％. The average recoveries were 99.40％-100.89％（RSDs were 0.18％-0.49％，n＝3）. The contents of sinapine thiocyanate in 3 batches of Shiwei yipi granules were 0.086 4-0.220 6 mg/g，the average was 0.168 4 mg/g. CONCLUSIONS ：The identification method of Massa Medicata Fermentata ，Galli Gigerii Endothelium Corneum ，Crataegi Fructus and Semen Raphani in Shiwei yipi granules as well as the method for content determination of sinapine thiocyanate in Semen Raphani are established successfully. The content of Semen Raphani in the Shiwei yipi granules is no less than 0.16 mg/g calculated by sinapine thiocyanate （C16H24NO·5 SCN）.

Objective:To investigate the application of different imaging methods of 99Tc m-pyrophosphate (PYP) in the diagnosis and pathological classification of cardiac amyloidosis (CA). Methods:A total of 31 patients (22 males, 9 females, age 21-81(57.2±13.4) years) with suspected CA who underwent 99Tc m-PYP dual-phase scintigraphy (early-phase: 1 h, delay-phase: 2-3 h) and SPECT/CT (1 h) between December 2018 and December 2019 in Peking Union Medical College Hospital were retrospectively included. Taking clinical diagnosis as the standard, the results of visual score (≥2, positive) and semi-quantitative values (heart to contralateral lung (H/CL)≥1.5, positive) of 99Tc m-PYP uptake in dual-phase scintigraphy and SPECT/CT imaging were analyzed. One-way analysis of variance and Bonferroni test were used to analyze the data. Results:Among 31 patients with suspected CA, 15 were clinically diagnosed as CA (5 patients with transthyretin-related CA (ATTR-CA) and 10 patients with light chain CA (AL-CA)) and 16 were diagnosed as non-CA. All 5 patients with ATTR-CA had positive dual-phase scintigraphy and SPECT/CT imaging results. Three out of 10 patients with AL-CA had positive early-phase scintigraphy whereas negative delay-phase scintigraphy and SPECT/CT imaging results. Sixteen patients who were clinically diagnosed as non-CA had negative dual-phase scintigraphy and SPECT/CT imaging results. The sensitivity (5/5), specificity (10/10), positive predictive value (5/5), negative predictive value (10/10) and accuracy (15/15) of delay-phase scintigraphy and SPECT/CT imaging were the same. Among 31 patients, 16 patients carried transthyretin-related (TTR) gene mutation, and 4 of them who clinically diagnosed as variant ATTR (ATTRv) had positive image findings while 12 of them who not clinically diagnosed as CA had negative image findings. There were significant differences in H/CL between ATTR-CA group and AL-CA group in early-phase (2.11±0.24 vs 1.31±0.07) and delay-phase (2.02±0.19 vs 1.30±0.05; F values: 75.41 and 87.15, Bonferroni test, both P<0.01). Conclusions:99Tc m-PYP delay-phase scintigraphy and SPECT/CT have high diagnostic efficiencies in ATTR-CA, helping to determine the pathological classification of CA; while early-phase scintigraphy has false positive results. Moreover, 99Tc m-PYP imaging is helpful to detect CA in patients with TTR gene mutation.