Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody.

Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.

Rapid detection of immunity against bacteria in Asian honeybee and Western honeybee with quantification of royalisin in the hemolymphe by fast ELISA.

Royalisin from royal jelly (RJ) is a valuable peptide both for the prevention of honeybee diseases and for RJ preservation. ELISA for fast determination of royalisin content in hemolymphe (RCH) of honeybees with polyclonal antibody against recombinant royalisin from Asian honeybee was established. Assay on RCHs of health samples from Asian honeybee and Western honeybee showed the former (7.06 µg/mL) was significantly higher than that of the latter (5.64 µg/mL, p < 0.01). Moreover, relative to the non infection, the RCHs of Asian honeybees at 24 and 48 h post infection of Eschericha coli were higher than those of Western honeybees by 32.90% and 29.66%, respectively. Evidence revealed that Asian honeybee possesses higher innate immunity and immune response against bacteria in relation to the Western honeybee. The method will be a potential tool for detection of resistant levels to pathogens in honeybees and for quantification of royalisin in RJ products.

[Prevention and treatment of hepatitis B recurrence after liver transplantation].

OBJECTIVE: To investigate the effect of the regimen of lamivudine (LAM) combined with hepatitis B immunoglobulin (HBIG) in prevention and treatment of re-infection of hepatitis B virus (HBV) and recurrence of hepatitis B after orthotopic liver transplantation (OLT) for HBV related end stage liver disease. METHODS: The clinical data of 183 adult liver transplantation patients who lived more than 6 months and were followed up for 14.6 months with complete data were studied retrospectively. According to the HBV prevention strategy, these recipients were divided into two groups: group of pure LAM (n = 106) and group of LAM plus intramuscular injection of low dose HBIG (n = 77). RESULTS: The rate of HBsAg negative conversion 1 week after OLT of the LAM group was 82.10% (87/106), significantly lower than that of the LAM + HBIG group [94.81% (73/77), P = 0.010]. The rates of HBV reinfection, HB recurrence, and YMDD mutation of the lamivudine group were 16.98% (18/106), 11.32% (12/106), and 8.49% (9/106) respectively, all significantly higher than those of the LAM + HBIG group [6.49% (5/77), 2.60% (2/77), and 1.30% (1/77) respectively, P = 0.035, 0.028, and 0.035 respectively]. All the patients with YMDD mutation were treated with adefovir (ADF) with improvement. Analysis showed no obvious difference in the effect of LAM given intramuscularly or intravenously. CONCLUSION: The protocol of combination of LAM and HBIG is highly effective, safe, and cost-effective in preventing the recurrence of HBV after OLT. YMDD mutation can be treated by ADF with satisfactory results.