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OBJECTIVE：To compare the effects of raw pro duct and different processed products of Gekko gecko on kidney-yang deficiency model mice induced by adenine. METHODS ：Totally 100 mice were randomly divided into blank group （n＝10）and modeling group （n＝90）. Modeling group was given adenine （50 mg/kg）intragastrically for 10 days to induce kidney-yang deficiency model ；blank group was given normal saline （0.2 mL/10 g）intragastrically. After modeling ，70 mice were randomly divided into model group ，positive group （Jinkui shenqi pill ，6.4 g/kg），G. gecko crude product group （1.2 g/kg）， wine-processed G. gecko group（1.2 g/kg）and oil-processed G. gecko group（1.2 g/kg）according to body weight and symptoms of kidney-yang deficiency ，with 14 mice in each group. Blank group and model group were given constant volume of normal saline intragastrically；administration groups were given relevant medicine intragastrically （0.2 mL/10 g），once a day ，for consecutive 14 d. During the experiment ，the symptoms and signs of mice in each group were observed. After last medication ，renal index ，testis index and serum levels of T ，CORT，BUN and Cr were measured ；HE staining method was used to observe the pathological changes of renal tissue of mice in each group. RESULTS ：Compared with blank group ，the mice in the model group suffered from performance of kidney-yang deficiency ，such as weight loss ，crouch and arch back ，chills and cold limbs ，and sparse body hair ， while renal index and serum levels of BUN and Cr were increased significantly （P＜0.01）. In renal tissue ，there were BA28117 pathological damages such as irregular arrangement of renal KY2016YB211tubular epithelial cells ，light staining of nucleus and edema of cytoplasm. Compared with model group ， performance of kidney-yang deficiency was improved to different extents in G. gecko crude product group and processed product groups（especially in wine-processed G. gecko group）；serum levelsof BUN and Cr were decreased significantly （P＜0.05 or P＜0.01）；pathological damage of renal tissue was alleviated in different degrees. In addition ，body weight of mice was increased significantly in G. gecko processed products groups （P＜0.01），and renal indexes of mice were decreased significantly in G. gecko crude product group and processed products groups （P＜0.05 or P＜0.01）. Compared with G. gecko crude product group ，renal index ，serum levels of BUN and Cr were significantly decreased in wine-processed G. gecko group（P＜0.01），and serum level of Cr was significantly decreased in oil-processed G. gecko group（P＜ 0.05）. CONCLUSIONS ：G. gecko crude product ，wine-processed G. gecko and oil-processed G. gecko all show a certain improvement effect on kidney-yang deficiency mice induced by adenine ，especially wine-processed G. gecko .
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Gekko gecko (Tokay Gecko) is a valuable traditional Chinese medicine. In this study, the loop-mediated isothermal amplification (LAMP) technique was introduced for visual rapid identification of G. gecko from adulterants. A total of sixty-five 12S rRNA sequences of fourteen species of G. gecko and its adulterants were obtained. The results showed that G. gecko could be identified from its adulterants through BLAST analysis based on 12S rRNA regions. The 12S rRNA sequences of ten batches of G. gecko were conserved. There were only two haplotypes and three variation sites in the available regions for primers design. Six specific LAMP primers were successfully designed online based on 12S rRNA sequences. The visual rapid detection of G. gecko could be achieved with the optimized conditions (64 °C for 1 h and 80 °C for 5 min). And the required minimal template concentration was 5 μg·L⁻¹ while conventional PCR with 0.5 mg·L⁻¹. Consequently, the LAMP method established from this study was rapid, specific, highly sensitive, and simple. It could be applied to detect G. gecko from its adulterants efficiently.
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DNA barcoding technology is widely opplied for species identification and discovery by using short and standard fragments of DNA sequences. In this study, the cytochrome c oxidase subunit 1 (COI) sequence as a DNA barcode was used to identify the Gekkogecko Linnaeus medicinal materials and adulterants in order to provide a new identification method of G. gecko Linnaeus. Genomic DNA was extracted from the experimental samples. The COI regions of nrDNA were amplified using polymerase chain reaction and sequenced bidirectionally. The alignment and NJ tree construction were performed in MEGA6.0. COI sequences can be obtained from all samples. The average intra-specific K2P distance of G. gecko Linnaeus was 0.005 and the maximum intra-specific distance was 0.013. All the G. gecko Linnaeus medicinal samples clustered into a clade in the NJ tree and can be distinguished from the adulterants. In a conclusion, COI can be used to correctly identify G. gecko Linnaeus, and it will be a potential DNA barcode for identification of other animal medicinal materials.
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Objective:To study the effects of the Gekko gecko ethanol extract on granular cells apoptosis of rat ovaries.Methods :3 month and 6 month female Wistar rats were taken in this study, and 20 rats were in each age group.Each age group were randomly divided into two groups(experimental group and control tragastric administration in experimental group, corresponding volume of 0.9% sodium chloride was given in control group by the same way.Apoptosis of granular cells and distribution of cell cycle in rat ovary were measured by terminal-deoxynucleotidyl transferase mediated d-UTP nick end labeling (TUNEL) and propidi-um iodide(PI) staining, AnnexinV/PI double labelling staining by flow cytometry (FCM) after administration for 15 days.Re8utts:The apoptosis numbers of granular cells in 3 month and 6 month age experimental group were 34 ±7 and 53 ±11, in control group were 48±9 and 76±17 respectively by TUNEL.There was signrficantly statistic difference (P <0.01).The apoptosis rates in 3 month and 6 month age experimental group were(6.06 ±2.01)% and (12.16 ±3.26) % , in the control group were (8.23±2.13) % and (23.69 ±6.28)% respectively by PI staining.There was significantly statistic drfference (P <0.05).However, no significant difference was found between two groups in cellular cycle distribution (P > 0.05).Early apoptosis rates in 3 month and 6 month age experimental group were (6.71±3.11) % and (23.74±6.28) % , in the control group were (19.05 ±5.78) % and (36.55± 8.35) % respectively by AnnexinV/PI double labelling staining FCM.There was significantly statistic difference (P <0.01).The apoptosis of granular cells were sig-nificantly less in experimental groups comparing wrth control groups both in 3 month and 6 month age group (P<0.01 or P<0.05).Conclusions :The ethanol extract of gekko gecko can inhibit the apoptosis of granu-lar cells in rat ovary, then improves the ovary function and delay ageing of rat.
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Objective To determine the content of cholesterol in Gekko Gecko.Methods High performance liquid chromatography was performed on Hypersil ODS2 (4.6 mm?150 mm,5 ?m).The chromatorgraphic conditions were as follows:methanol as mobile phase,flow rate being 1.0 mL/min and detecting wavelength at 208 nm.The column temperature was 40 ℃.Results A good linearity of cholesterol was shown in range of 0.885～14.000 ?g (r=0.999 5),the average recovery was 99.65%,RSD=3.45% (n=6).Conclusions This method is simple,rapid and accurate,and may provide a reliable way for the quantification of cholesterol in Gekko Gecko.
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The effects of alcoholic extract of Gekko gecko L. on the activities of SOL), GSH-Px, CAT and the level of GSH and LPO in the rat gut were investigated. The data presented in this paper indicated that the clcrance of free radical was accelerated, LPO was decreased, the activities of SOD, GSH-Px and CAT was elevated, and, the content of GSH was increased. The extract from the tail part of the Gekko gecko L. was more active than the extract from the body. These results and our other results together indicate that Gekko gecko L. has an obvious antiaging effect
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Objective To clarify the position,figure and connections with adjacencies in the pallial thickening(Pth),and provide essential parameters for its function study. Methods The coronal serial sections of 60?m thickness in gekko gecko brain were made by cryo-microtome,and Nissl staining was used.Pictures were taken in each coronal section containing the Pth and the size of Pth in each section was measured.One of them was chosen for the three-dimensional reconstruction.3D MAX was used as the tool software to rebuild the nucleus. Results 1.The Pth was located in the rostral part of the telencephalon,the lateral part of anterior dorsal ventricular ridge,the medial part of the lateral cortex and the ventral part of the dorsal cortex.The length of Pth from the rostral to the caudal end was(912.67?110.96)?m(n=10),the cubage of Pth was about(0.1430?0.0414)?m~3(n=10).2.The Pth could be divided into four segments,the anterior,the middle,the posterior and the terminal segments from the rostral to the caudal end.In the posterior segment,its dorsoventral axis was the longest,and could be divided into two parts: the dorsal and the ventral parts.The boundary of the two parts was clear.Conclusion The Pth is a long,narrow and flat structure;its rostrocaudal axis is longer than its dorsoventral axis,and its dorsal edge is smoother than its ventral edge.In the Pth,its caudal region is larger than its rostral region,and the posterior segment in the caudal region is divided into the dorsal and the ventral cell populations.