RESUMO
ObjectiveTo explore the scientific connotation of fried charcoal survivability of Lonicerae Japonicae Flos（LJF） by analyzing the correlation between the color change and the intrinsic components during the processing of LJF Carbonisata（LJFC）， and taking pH， charcoal adsorption and microscopic characteristics as indexes. MethodLJFC samples with different degrees of processing were prepared according to the stir-frying time of 0.0， 1.5， 3.0， 4.5， 6.0， 7.5， 9.0， 10.5 min（numbered S1-S8）， and the contents of gallic acid， chlorogenic acid， cryptochlorogenic acid， rutin， luteoloside， isochlorogenic acid A and isochlorogenic acid C were determined by high performance liquid chromatography（HPLC）， and the L*（brightness）， a*（red-greenness） and b*（yellow-blueness） of LJFC samples with different degrees of processing were determined by spectrophotometer， and the correlation analysis and principal component analysis（PCA） between the contents of seven representative components and the color of the samples were carried out by SPSS 26. 0 and SIMCA-P 14.1. Then pH， adsorption force and characteristic structure of different samples of LJFC were detected and the processing pattern of LJFC was analyzed. ResultThe results of quantitative analysis revealed that the contents of luteoloside， rutin， chlorogenic acid and isochlorogenic acid A gradually decreased， and the contents of cryptochlorogenic acid， isochlorogenic acid C and gallic acid firstly increased and then decreased. The L* and b* of the sample powders decreased， and a* showed a trend of increasing and then decreasing. The L* and b* were positively correlated with the contents of chlorogenic acid， rutin， luteoloside， isochlorogenic acid A， b* was positively correlated with the content of gallic acid， and a* was positively correlated with the contents of cryptochlorogenic acid and isochlorogenic acid C. PCA revealed that samples could be clearly divided into 3 groups， S1-S2 as one group， S3-S5 as one group， and S6-S8 as one group， with S3 having the highest score. The results of regression analysis showed that only isochlorogenic acid C could be used to predict the contents of components by colorimetric values combined with regression equations. Physicochemical analysis showed that pH of LJFC increased with the increase of degree of charcoal stir-frying， while adsorption force showed a tendency of increasing and then decreasing， with the highest adsorption force in the S5 sample， and the non-glandular hairs， calcium oxalate clusters and pollen grains had a varying degree of decreasing with the deepening of processing degree， and the microstructures of S6-S8 samples were obviously charred with pollen grains almost invisible. ConclusionThe changes in chemical composition and color characteristics of LJFC during the processing have certain correlations， combined with the changes in physicochemical properties， S5 sample is found to be the optimal processed products， which can provide a reference for the processing standardization and quality evaluation of LJFC， and enrich the scientific connotation of fried charcoal survivability of LJF.
RESUMO
Objective Autoimmune inner ear disease is one of the inner ear diseases that can be effectively treated clinically, and researches on it have significant clinical and practical value.This study explored the immunoregulating and therapeutic effects of the mi-croRNA-146a recombinant lentiviral vector (RLV) on immune-medicated inner ear disease (IMIED) in guinea pigs so as to avoid the adverse reactions of the currently used immunodepressants. Methods IMIED models were established in 30 guinea pigs by immuni-zation with keyhole limpet hemocyanin and divided into three groups: microRNA-146a RLV, empty lentiviral vector ( ELV) control, and surgery simulation control, microinjected via the scala tympani with microRNA-146a RLV, ELV, and PBS, respectively.Before and after immunization and at 7 days after microinjection, the auditory function and the level of specific anti-KLH antibodies in the ser-um were measured by auditory brainstem response ( ABR) audiometry and ELISA, respectively.Seven days after microinjection, 3 ani-mals in each group were subjected to HE staining and light microscopy, another 3 to fluoro-autography for realizing the situations of the inner ear transfected by the lentiviral vector, and the other 4 to measurement of the contents of microRNA-146a in the inner ear tissue. Results Compared with the baseline, immunization significantly increased the level of specific anti-KLH antibodies in the serum (0.09 ±0.01 vs 1.90 ±0.74 in the RLV group, 0.11 ±0.02 vs 2.20 ±0.75 in the ELV group, and 0.11 ±0.02 vs 2.10 ±0.64 in the surgery simulation control) as well as the average threshold of the ABRⅢwave (left ear 11.67 ±2.58 vs 61.67 ±5.16 and right ear 12.50 ±2.73 vs 60.00 ±4.47 in the RLV group;left ear 14.16 ±3.76 vs 64.33 ±9.17 and right ear 13.33 ±2.58 vs 60.83 ± 4.92 in the ELV group;left ear 15.83 ±3.76 vs 64.17 ±10.2 and right ear 15.00 ±5.47 vs 62.50 ±9.35 in the surgery simulation control) .The average threshold of the ABRⅢwave was decreased after local injection into the inner ear as compared with that after immunization, with no statistically significant difference between the ELV and surgery simulation control groups.Fluoro-autographic ob-servation showed that the main parts of the inner ear transfected with microRNA-146a recombinant lentiviruses were the spiral limbus, spiral ganglion afferent fibers, basal cells of the spiral ligament, Corti organ, and psalterial cords.The immune inflammatory reaction of the inner ear was significantly reduced in the RLV group as compared with the ELV and surgery simulation control groups, while the content of microRNA-146a in the inner ear tissue was obviously increased in the former group than in the latter two. Conclusion The microRNA-146a recombinant lentiviral vectors are widely distributed and transfected in the inner ear tissue after injected via the scala tympani.Injecting recombinant microRNA-146a lentiviral vectors into the inner ear can significantly reduce pathological damage and auditory dysfunction caused by inner ear immune inflammatory reaction.