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ObjectiveOn the basis of sensory evaluation， the changes of volatile components in gecko before and after processing were compared， and the odor correction effect of different processing methods of gecko was discussed. MethodRaw products， fried yellow products， vinegar processed products， wine processed products， talcum powder scalding products and white wine sprayed products after scalding talcum powder of gecko were prepared， and 10 odor assessors were invited to evaluate the 6 samples in turn by sensory evaluation. Headspace solid-phase microextraction-gas chromatography-mass spectrometry （HS-SPME-GC-MS） and relative odor activity value （ROAV） were used to analyze the key odor components， and multivariate statistical methods were used to analyze the difference of volatile components between raw and processed products of gecko. Taking water-soluble extract and protein contents as internal indicators， sensory evaluation score and content ranking of differential components as external indicators， and assigning a weight of 0.25 to them respectively， the comprehensive scores of raw products and processed products of gecko were calculated to evaluate the odor correction effect of each processing method. ResultThe average sensory evaluation scores of the raw products， fried yellow products， vinegar processed products， wine processed products， talcum powder scalding products and white wine sprayed products after scalding talcum powder of gecko were 1.6， 5.2， 6.2， 6.1， 7.2 and 8.0， respectively. ROAV results showed that key components affecting odor of gecko were 2-ethyl-3，5-dimethylpyrazine， isovaleraldehyde， trimethylamine， 1-octen-3-ol， n-octanal， nonanal， 2-methylnaphthalene， γ-octanolide， 2-heptanone and phenol. Principal component analysis （PCA） significantly distinguished raw products from processed products. Orthogonal partial least squares-discriminant analysis （OPLS-DA） results showed that there were 16， 13， 16， 16， 16 differential components between raw products， fried yellow products， vinegar processed products， wine processed products， talcum powder scalding products and white wine sprayed products after scalding talcum powder of gecko. Among these differential components， there were 4 common components， namely， the contents of different odor components （2-methylnaphthalene and 2-ethyl-p-xylene） decreased， while the contents of different flavor components （2-decanone and 2，3，5-trimethylpyrazine） increased. The comprehensive scoring results showed that the odor correction effect of each processed products was in the order of talcum powder scalding products>wine processed products>vinegar processed products>fried yellow products>white wine sprayed products after scalding talcum powder. ConclusionTalcum powder scalding is a better method to improve the odor of gecko， and it can provide an experimental basis for the processing of gecko to correct the odor.
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BACKGROUND@#Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs).@*METHODS@#Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited.@*RESULTS@#The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs.@*CONCLUSION@#Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.
Subject(s)
Humans , Apoptosis , Autophagy , Eccrine Glands , Epithelial Cells , OrganoidsABSTRACT
OBJECTIVE：To compare the difference in the antioxid ant effect of fresh Rehmannia glutinosa ，dried R. glutinosa ， R. glutinosa preparata during ancient characteristic processing and its polysaccharides before and after processing on aging model rats，and to provide reference for the processing of R. glutinosa . METHODS ：The sample of R. glutinosa preparata was prepared according to ancient characteristic method. During the processing ，the fresh and dried R. glutinosa samples were retained. Then crude polysaccharide were extracted from fresh R. glutinosa and Rehmanniae radix preparata by water extraction and alcohol precipitation. Totally 96 rats were divided into blank group （water），model group （water），positive control group [vitamine C ，100 mg/（kg·d）]，fresh R. glutinosa group [ 700 mg/（kg·d）]，dried R. glutinosa group [ 135 mg/（kg·d）] ，Rehmanniae radix preparata group [ 135 mg/（kg·d）]，fresh R. glutinosa polysaccharide group [ 1 400 mg/（kg·d），by the weight of fresh R. glutinosa ] and Rehmanniae radix preparata polysaccharide group mg/（kg·d），by the weight of Rehmanniae radix preparata] ， with 12 rats in each group. Except for blank group ，other 制。E-mail：zhouyan1221@163.com groups were given D-galactose [ 125 mg/（kg·d）] on neck and back to induce sub-acute aging model. At the same time ，they were given relevant medicine in tragastrically，once a day ，for consecutive 56 days. After last admin istration，the liver ，brain，kidney，spleen，heart and thymus indexes were determined. The total antioxidant capacity （T-AOC），superoxide dismutase （SOD）activity，catalase（CAT）activity and MDA content in serum ， liver，brain and kidney were determined. RESULTS ：Compared with blank group ，organ indexes of rats in the model group were decreased significantly （P＜0.05 or P＜0.01）；T-AOC，SOD activity and CAT activity in serum ，brain，liver and kidney tissue were decreased significantly （P＜0.01），while MDA content increased significantly （P＜0.01）. Compared with model group ，the organ indexes of brain ，liver and kidney ，SOD activity in serum and kidney of fresh R. glutinosa group were not significantly increased （P＞0.05）；kidney index ，T-AOC in serum and brain ，SOD activity in serum ，liver and kidney tissue were not significantly increased in the dried R. glutinosa group（P＞0.05）；kidney index ，T-AOC in serum and cerebral tissue ，SOD activity in serum were not significantly increased in fresh R. glutinosa group（P＞0.05）；other organ indexes ，T-AOC，SOD activity and CAT activity in serum and tissues were increased significantly in other groups （P＜0.05 or P＜0.01），while MDA content in serum and tissues were decreased significantly in all administration groups （P＜0.05 or P＜0.01）. Compared with fresh R. glutinosa group，T-AOC in serum was decreased significantly in dried R. glutinosa group（P＜0.01），and there was no significant difference in other indexes （P＞0.05）；kidney and spleen indexes of rats in Rehmanniae radix preparata group were increased significantly （P＜0.05），T-AOC in renal tissue ，SOD activity in serum ，cerebral tissue and renal tissue ，CAT activity in cerebral and liver tissue were increased significantly （P＜0.05 or P＜0.01），while MDA in cerebral and liver tissue were significantly decreased （P＜ 0.01）. CAT in cerebral tissue and liver tissue of rats in Rehmanniae radix preparata group were significantly higher than those in positive control group （P＜0.01）. Compared with fresh R. glutinosa polysaccharide group ，spleen and renal indexes of rats in Rehmanniae radix preparata group were increased significantly （P＜0.05 or P＜0.01），T-AOC，SOD activity and CAT activity in serum and cerebral ，liver，renal tissues were increased significantly （P＜0.05 or P＜0.01）. T-AOC and CAT activity of cerebral ， liver and renal tissues in Rehmanniae radix preparata group were all significantly higher than those in positive control group （P＜ 0.05 or P＜0.01）. CONCLUSIONS ：In the aspect of increasing organ index and improving the activity of antioxidant enzymes in serum，cerebral，liver and
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OBJECTIVE：To establish the fingerprint of raw produc ts and different processed products of Pinellia pedatisecta ， to determine the contents of 5 kinds of nucleosides ，and to compare the differences of components between the raw products and processed products. METHODS ：P. pedatisecta raw products ，processed products by Processing Standard of Chinese Medicine in Henan Province （called“Standard processed product ”for short ）and processed products by new integrated processing technology in the production area （called“new integrated processed product ”for short ）were collected as investigation objects （12 batches of each）. The determination was performed on SymmetryShield RP 18 column with mobile phase consisted of acetonitrile （A）-0.1％ acetic acid aqueous water solution （B）（gradient elution ）at the flow rate of 0.8 mL/min，with the column temperature of 30 ℃， the detection wavelength of 270 nm，and the injection volume of 15 µL. HPLC fingerprints of 3 kinds of P. pedatisecta samples were established by using Similarity Evaluation System of TCM Chromatographic Fingerprints （2004 A version ） ，and the similarity of fingerprints was evaluated. The chromatographic peaks were identified by comparing with the reference chromatogram. Five nucleosides （adenine，hypoxanthine，uridine，xanthine，inosine）were quantitatively analyzed. SPSS 21.0 software was used for cluster analysis of 36 batches of samples. RESULTS ：The results of fingerprint and content determination met the relevant requirements. The similarity of 3 kinds of sample with their control fingerprint were all greater than 0.990. There were 22 common peaks in the raw products of P. pedatisecta ，and 16 common peaks were identified in the 2 kinds of processed products （the same 6 peaks disappeared from 2 kinds of processed products ）. Fivecomponents were identified in 3 kinds of samples ，such as adenine（peak 3），hypoxanthine（peak 7），uridine（peak 8）， 1064056472@qq.com xanthine（peak 9）and inosine （peak 11）. Results of content determination showed that total contents of 5 kinds of nucleosides in 2 kinds of proc essed products were all· decreased；the contents of them in descending order w as raw product ＞new i ntegrated processed products ＞Standard processed products. Results of cluster analysis showed that 36 batches of samples could be clustered into 2 categories，i.e. raw product was clustered into one category and 2 kinds of processed products into other one . CONCLUSIONS ：Established method is stable ， feasible and suitable for the quality evaluation of raw products and different processed products of P. pedatisecta . Fingerprints have changed significantly and the total content of 5 kinds of nucleosides in P. pedatisecta are all decreased after processing ，but that of new integrated processed products is slightly higher than that of Standard processed products .