1.Study on the Effect of Regulating DHPR/RyR Pathway by Pressing and Rubbing Method on the Improvement of Myofascial Pain Syndrome in Rats
Chao XIANG ; Sheng-Hua HE ; Xin ZHAO ; Qi WAN ; Chi MA ; Yan-Ping HU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(5):1270-1276
Objective To explore the therapeutic effect and mechanism of pressing and rubbing method on myofascial pain syndrome(MPS)rats.Methods A total of 12 rats were randomly selected from 60 rats as the normal group,and the remaining rats were used to construct the MPS model by blunt strike combined with centrifugal exercise.Then 48 successfully modeled rats were randomly divided into model group,pressing and rubbing method group,pressing and rubbing method + Dantrolene[ryanodine receptor(RyR)inhibitor]group,pressing and rubbing method + normal saline group,with 12 rats in each group.The mechanical pain threshold was measured by von-Frey method.Detection of soft tissue tension,electromyography was performed;the ultrastructure of the pain point tissue was observed by transmission electron microscopy.The content of calcium ion(Ca2+)in the tissue of trigger point was detected by colorimetry.The protein expressions of dihydropyridine receptor(DHPR)α1,RyR and acetylcholinesterase(AChE)in the pain points of rats were detected by Western Blot.Results Compared with the normal group,the mechanical pain threshold,soft tissue tension in trigger point and the protein expressions of DHPRα1,RyR and AChE in the model group were decreased and the Ca2+ + content was increased(all P<0.05),and the peak potential with higher amplitude was observed in the electromyogram.The ultrastructure of the trigger point tissue was damaged.Compared with the model group,the mechanical pain threshold,soft tissue tension of trigger point and the protein expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method group and the pressing and rubbing method + normal saline group were increased,and the Ca2+ content was decreased(all P<0.05),the electromyography was restored to be stable,the ultrastructural damage of the trigger point tissue was alleviated.Compared with the pressing and rubbing method group,the mechanical pain threshold,soft tissue tension of trigger point and protein the expressions of DHPRα1,RyR and AChE in the trigger point tissue of the rats in the pressing and rubbing method + Dantrolene group were decreased,and the Ca2+ content was increased(all P<0.05),the electromyogram showed electrical activity changes,the ultrastructure of the trigger point tissue was damaged.Conclusion The pressing and rubbing method may improve MPS in rats by activating the DHPR/RyR signaling pathway.
2.Experimental study on concentration selection of immunohistochemical antibodies for retinal endoplasmic reticulum stress
Xiao-Hong CHEN ; Wan-Jiao LIANG ; Shi-Shu HUANG ; Yan SUN ; Xin LUO ; Lu LAI ; Zhao-Sheng CHI ; Mei-Zhu CHEN ; Yun-Peng WANG ; Wei-Ming YAN
International Eye Science 2023;23(1):32-38
AIM: To explore the optimal concentration of endoplasmic reticulum stress immunohistochemical(IHC)staining antibody in mouse retinitis pigmentosa(RP)model, which provides the corresponding index detection method for studying the pathogenesis and intervention measures of RP.METHODS: Clean male C57BL/6J mice were intraperitoneally injected with N-methyl-N-nitrosourea(MNU, 60mg/kg)to prepare RP mouse model. Electroretinogram(ERG)and hematoxylin-eosin(HE)staining were performed on 7d after modeling to verify the successful modeling. The expression of endoplasmic reticulum stress-related proteins(IRE1, ATF6, PERK, GRP78, Caspase-12)was detected by IHC staining.RESULTS: The following proteins, including IRE1, ATF6, PERK, GRP78 and Caspase-12, were positively expressed in retina of RP mouse. The optimal concentrations of the above proteins were as follows: IRE1 antibody concentration was 1:1000, ATF6 antibody concentration was 1:500 and 1:1000(with no difference in positive expression, P>0.05), PERK antibody concentration was 1:1500, GRP78 antibody concentration was 1:200 and Caspase-12 antibody concentration was 1:100, the proteins were well expressed at the above concentrations, and the positive expressions of corresponding proteins were different from those of other antibody concentrations(P<0.05).CONCLUSION: The optimal concentrations for IHC staining in different proteins of mouse RP models were as follows: the concentrations of endoplasmic reticulum stress-related protein antibodies were 1:1000 in IRE1, 1:500 and 1:1000 in ATF6, 1:1500 in PERK, 1:200 in GRP78, and 1:100 in Caspase-12.
3.Anatomical and histological investigation of the area anterior to the anorectum passing through the levator hiatus.
Xiao Jie WANG ; Zhi Fang ZHENG ; Qian YU ; Wen LI ; Yu DENG ; Zhong Dong XIE ; Sheng Hui HUANG ; Ying HUANG ; Xiao Zhen ZHAO ; Pan CHI
Chinese Journal of Gastrointestinal Surgery 2023;26(6):578-587
Objective: To document the anatomical structure of the area anterior to the anorectum passing through the levator hiatus between the levator ani slings bilaterally. Methods: Three male hemipelvises were examined at the Laboratory of Clinical Applied Anatomy, Fujian Medical University. (1) The anatomical assessment was performed in three ways; namely, by abdominal followed by perineal dissection, by examining serial cross-sections, and by examining median sagittal sections. (2) The series was stained with hematoxylin and eosin to enable identification of nerves, vessels, and smooth and striated muscles. Results: (1) It was found that the rectourethralis muscle is closest to the deep transverse perineal muscle where the longitudinal muscle of the rectum extends into the posteroinferior area of the membranous urethra. The communicating branches of the neurovascular bundle (NVB) were identified at the posterior edge of the rectourethralis muscle on both sides. The rectum was found to be fixed to the membranous urethra through the rectourethral muscle, contributing to the anorectal angle of the anterior rectal wall. (2) Serial cross-sections from the anal to the oral side were examined. At the level of the external anal sphincter, the longitudinal muscle of the rectum was found to extend caudally and divide into two muscle bundles on the oral side of the external anal sphincter. One of these muscle bundles angled dorsally and caudally, forming the conjoined longitudinal muscle, which was found to insert into the intersphincteric space (between the internal and external anal sphincters). The other muscle bundle angled ventrally and caudally, filling the gap between the external anal sphincter and the bulbocavernosus muscle, forming the perineal body. At the level of the superficial transverse perineal muscle, this small muscle bundle headed laterally and intertwined with the longitudinal muscle in the region of the perineal body. At the level of the rectourethralis and deep transverse perineal muscle, the external urethral sphincter was found to occupy an almost completely circular space along the membranous part of the urethra. The dorsal part of the external urethral sphincter was found to be thin at the point of attachment of the rectourethralis muscle, the ventral part of the longitudinal muscle of the rectum. We identified a venous plexus from the NVB located close to the oral and ventral side of the deep transverse perineal muscle. Many vascular branches from the NVB were found to be penetrating the longitudinal muscle and the ventral part of rectourethralis muscle at the level of the apex of the prostate. The rectourethral muscle was wrapped ventrally around the membranous urethra and apex of the prostate. The boundary between the longitudinal muscle and prostate gradually became more distinct, being located at the anterior end of the transabdominal dissection plane. (3) Histological examination showed that the dorsal part of the external urethral sphincter (striated muscle) is thin adjacent to the striated muscle fibers from the deep transverse perineal muscle and the NVB dorsally and close by. The rectourethral muscle was found to fill the space created by the internal anal sphincter, deep transverse perineal muscle, and both levator ani muscles. Many tortuous vessels and tiny nerve fibers from the NVB were identified penetrating the muscle fibers of the deep transverse perineal and rectourethral muscles. The structure of the superficial transverse perineal muscle was typical of striated muscle. These findings were reconstructed three-dimensionally. Conclusions: In intersphincteric resection or abdominoperineal resection for very low rectal cancer, the anterior dissection plane behind Denonvilliers' fascia disappears at the level of the apex of the prostate. The prostate and both NVBs should be used as landmarks during transanal dissection of the non-surgical plane. The rectourethralis muscle should be divided near the rectum side unless tumor involvement is suspected. The superficial and deep transverse perineal muscles, as well as their supplied vessels and nerve fibers from the NVB. In addition, the cutting direction should be adjusted according to the anorectal angle to minimize urethral injury.
Humans
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Male
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Rectum/surgery*
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Anal Canal/anatomy & histology*
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Rectal Neoplasms/surgery*
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Proctectomy
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Urethra/surgery*
4.Morinda officinalis oligosaccharides increase serotonin in the brain and ameliorate depression via promoting 5-hydroxytryptophan production in the gut microbiota.
Zheng-Wei ZHANG ; Chun-Sheng GAO ; Heng ZHANG ; Jian YANG ; Ya-Ping WANG ; Li-Bin PAN ; Hang YU ; Chi-Yu HE ; Hai-Bin LUO ; Zhen-Xiong ZHAO ; Xin-Bo ZHOU ; Yu-Li WANG ; Jie FU ; Pei HAN ; Yu-Hui DONG ; Gang WANG ; Song LI ; Yan WANG ; Jian-Dong JIANG ; Wu ZHONG
Acta Pharmaceutica Sinica B 2022;12(8):3298-3312
Morinda officinalis oligosaccharides (MOO) are an oral drug approved in China for the treatment of depression in China. However, MOO is hardly absorbed so that their anti-depressant mechanism has not been elucidated. Here, we show that oral MOO acted on tryptophan → 5-hydroxytryptophan (5-HTP) → serotonin (5-HT) metabolic pathway in the gut microbiota. MOO could increase tryptophan hydroxylase levels in the gut microbiota which accelerated 5-HTP production from tryptophan; meanwhile, MOO inhibited 5-hydroxytryptophan decarboxylase activity, thus reduced 5-HT generation, and accumulated 5-HTP. The raised 5-HTP from the gut microbiota was absorbed to the blood, and then passed across the blood-brain barrier to improve 5-HT levels in the brain. Additionally, pentasaccharide, as one of the main components in MOO, exerted the significant anti-depressant effect through a mechanism identical to that of MOO. This study reveals for the first time that MOO can alleviate depression via increasing 5-HTP in the gut microbiota.
5.Consensus on clinical management of tumor-induced osteomalacia.
Yan JIANG ; Xiang LI ; Li HUO ; Yong LIU ; Wei LYU ; Lian ZHOU ; Wei YU ; Huan-Wen WU ; Xiao-Ping XING ; Mei LI ; Ou WANG ; Yue CHI ; Rui-Zhi JIAJUE ; Yu PEI ; Jian-Min LIU ; Jian-Ming BA ; Qiao ZHANG ; Zhi-Feng SHENG ; Zhen-Lin ZHANG ; Jia-Jun ZHAO ; Salvatore MINISOLA ; Wei-Bo XIA
Chinese Medical Journal 2021;134(11):1264-1266
6.Russian Characteristic Medicinal Plant Resources Based on DNA Barcode Technology
Ling-long LUO ; Chi ZHANG ; Bo DENG ; Bao-sheng ZHAO ; Yao-sheng ZHANG ; Yuan-yang SHAO ; Xiao-yan JIN ; Xue-yong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2020;26(2):175-180
Objective::To investigate the distribution status of medicinal plants in the wild areas of Russian Caucasus and Altai, and clarify the types and efficacy information of medicinal plants in the area, so as to dig deep into new resources and new functions of medicinal plants in the countries along the Belt and Road. Method::Medicinal plants in the wild were searched and collected to make waxy specimens, and sent back to the country to extract the total DNA of the leaves of the leaves. Internal Transcribed Spacer(ITS)sequence universal primers were used for Polymerase Chain Reaction (PCR)amplification. The PCR products were sent for the two-way sequencing, and the sequencing results are spliced by software according to National Center for Biotechnology Information(NCBI). The same ITS sequence of the highest similarity species obtained by database BLAST was analyzed by DNAman software to identify the ITS sequence of the species and the ITS sequence of the same species. The MEGA 7 software was used as the phylogenetic tree, and the Kimura-2 parameter genetic distance was used to construct the neighbor joining(NJ) phylogenetic tree by the neighbor-joining method. The confidence of each branch of the development tree was tested by the bootstrap test method. A total of 2 000 cycles were performed, and the results were identified based on the clustering results. On this basis, the key medicinal plants in the Russian Caucasus and Altay wild areas were summarized and analyzed. Result::After BLAST alignment in NCBI database, the ITS sequences of each specimen were clustered with the login sequences on the NCBI database, which were separated from the outer group. The species classification of the specimens to be identified was determined by combining the characteristics of the specimens. A total of 51 plants were identified from the specimens collected in the field, covering 44 genera of 17 families, and 29 plants had clear efficacy records. The National Drug List of the Russian Federation and the Chinese Pharmacopoeia were retrieved to summarize commonly used medicinal plants in China and conclude that 20 kinds of Chinese and Russian common medicinal materials have different medicinal effects in local areas. This study has a reference significance for expanding the scope and clinical experience of traditional Chinese medicines, and provides a basis for strengthened local species conservation, development and utilization of wild medicinal plant resources.
7.Adefovir Dipivoxil plus Chinese Medicine in HBeAg-Positive Chronic Hepatitis B Patients: A Randomized Controlled 48-Week Trial.
Xiao-Ke LI ; Ming-Xiang ZHANG ; Feng-Zhen SHAO ; Da-Qiao ZHOU ; Jing-Dong XUE ; Tie-Jun LIU ; Xiao-Ling CHI ; Bing-Jiu LU ; Xian-Bo WANG ; Qin LI ; Jun LI ; De-Wen MAO ; Hua-Sheng YANG ; Hong-Zhi YANG ; Wen-Xia ZHAO ; Yong LI ; Guo-Liang ZHANG ; Yi-Ming ZHAO ; Jian-Dong ZOU ; Meng-Yang LIU ; Ke-Ke ZHANG ; Xian-Zhao YANG ; Da-Nan GAN ; Ying LI ; Peng ZHANG ; Zhi-Guo LI ; Shuo LI ; Yong-An YE
Chinese journal of integrative medicine 2020;26(5):330-338
OBJECTIVE:
To evaluate the effects of a 48-week course of adefovir dipivoxil (ADV) plus Chinese medicine (CM) therapy, namely Tiaogan Jianpi Hexue () and Tiaogan Jiedu Huashi () fomulae, in hepatitis B e antigen (HBeAg)-positive Chinese patients.
METHODS:
A total of 605 HBeAg-positive Chinese CHB patients were screened and 590 eligible participants were randomly assigned to 2 groups in 1:1 ratio including experimental group (EG, received ADV plus CM) and control group (CG, received ADV plus CM-placebo) for 48 weeks. The major study outcomes were the rates of HBeAg and HBV-DNA loss on week 12, 24, 36, 48, respectively. Secondary endpoints including liver functions (enzymes and bilirubin readings) were evaluated every 4 weeks at the beginning of week 24, 36, and 48. Routine blood, urine, and stool analyses in addition to electrocardiogram and abdominal B scan were monitored as safety evaluations. Adverse events (AEs) were documented.
RESULTS:
The combination therapy demonstrated superior HBeAg loss at 48 weeks, without additional AEs. The full analysis population was 560 and 280 in each group. In the EG, population achieved HBeAg loss on week 12, 24, 36, and 48 were 25 (8.90%), 34 (12.14%), 52 (18.57%), and 83 (29.64%), respectively; the equivalent numbers in the CG were 20 (7.14%), 41 (14.64%), 54 (19.29%), and 50 (17.86%), respectively. There was a statistically significant difference between these group values on week 48 (P<0.01). No additional AEs were found in EG. Subgroup analysis suggested different outcomes among treatment patterns.
CONCLUSION
Combination of CM and ADV therapy demonstrated superior HBeAg clearance compared with ADV monotherapy. The finding indicates that this combination therapy may provide an improved therapeutic effect and safety profile (ChiCTR-TRC-11001263).
Adenine
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analogs & derivatives
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therapeutic use
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Adult
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Antiviral Agents
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therapeutic use
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Double-Blind Method
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Drug Therapy, Combination
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Drugs, Chinese Herbal
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therapeutic use
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Female
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Hepatitis B e Antigens
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immunology
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Hepatitis B, Chronic
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drug therapy
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immunology
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Humans
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Male
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Medicine, Chinese Traditional
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Organophosphonates
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therapeutic use
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Young Adult
8. Hypoglycemic Mechanism of Ziya Biti Tablet Based on LC-MS Metabolomics Technique
Cai-juan ZHANG ; Xiao-yan JIN ; Min-yi QIU ; Chi ZHANG ; Ling-long LUO ; Min WANG ; Bao-sheng ZHAO ; Xue-yong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(8):141-147
Objective: To observe the hypoglycemic effect of Uygur medicine Ziya Biti tablet on the type 2 diabetic rats, and analyze the hypoglycemic mechanism based on metabolomic techniques. Method: According to the results of clinical research about different origins of Ziya Biti tablet, the optimal composition was screened out; type 2 diabetic rats were taken as an experimental object in the pharmacodynamic experiments;the control group and model group were given the same dose of normal saline, Ziya Biti bablet group was given 300 mg·kg-1, the metformin group was given 300 mg·kg-1 metformin hydrochloride. The fasting blood and weight changes of the experimental group after the treatment were recorded and compared with normal group; ultra-high performance liquid chromatography-linear ion trap/electrostatic field orbit trap combined-type high resolution mass spectrometry (UHPLC-LTQ/Orbitrap MS) technology was used to conduct the metabolomics analysis on the rat serum, and principal component analysis (PCA), partial least squares discriminant analysis (OPLS-DA) on different groups of rat serum metabolites were performed to identify potential biomarkers. Result:Compared with the model group, the rats in the Uygur medicine Ziya Biti tablet showed a healthy states, and the blood glucose were decreased(P<0.05), which indicated that Uygur medicine Ziya Biti tablet had a certain hypoglycemic effect on type 2 diabetic rats. Eight biomarkers were finally obtained according to the results. Compared with model group, L-valine and propionylcarnitine in the Uygur medicine group were decreased(P<0.05), while sphingosine-1-phosphate, LPC (16:1/0:0), LPC (18:0/0:0), LPC (18:2/0:0), LPC 20:1/0:0), PC (19:0/0:0) were elevated(P<0.05). Conclusion:The experimental results showed that Uygur medicine Ziya Biti tablet can reduce the blood glucose of type 2 diabetic rats and allivate general physiological characteristics. The mechanism of action may be related to the improvement of amino acid metabolism and lipid metabolism.
9. Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells
Min WANG ; Ya-qi LI ; Quan-tao MA ; Chen-yue LIU ; Chi ZHANG ; Min-yi QIU ; Cai-juan ZHANG ; Ting WANG ; Bao-sheng ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):135-140
Objective: To observe effect of Mori Folium-containing serum on glucose consumption and cell activity of fat cell line 3T3-L1 insulin resistance (IR) model, in order to screen out the optimal concentration of drug-containing serum, detect effect of Mori Folium on the content of inflammatory factors, and explore the possible mechanism. Method: 3T3-L1 preadipocytes in logarithmic growth phase were selected, and induced with 10 mg·L-1 insulin (Ins), 0.25 mmol·L-1 dexamethasone (DEX) and 0.5 mmol·L-1 3-isobutyl-methylxanthine(IBMX) for 48 h and then with 10 mg·L-1 Ins for 48 h. After the cells were differentiated into mature adipocytes, they were induced with 1 μmol·L-1 DEX for 96 h to establish IR model. Glucose content in the supernatant of cells was detected by glucose oxidase after serum containing Mori Folium cultured for 12,24,36, 72 h. Methyl-thiazdyl-tetrazolium(MTT) was used to detect the effect of serum containing Mori Folium on IR cells activity. The content of tumor necrosis factor-α(TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, the effects of inflammatory factors on the expressions of insulin signaling pathway proteins insulin receptor (InsR), insulin receptor substrate (IRS), p-IRS1 and glucose transporter 4 (GLUT4) were determined by Western blot. Result: Serum containing Mori Folium could significantly increase the glucose consumption rate and cell activity of IR cells (P<0.01), reduce the content of TNF-α (P<0.01), and regulate the expression of insulin signaling pathway proteins (P<0.05,P<0.01). Conclusion: Mori Folium can significantly improve IR status of 3T3-L1 cells, and its mechanism may be related to inhibiting TNF-α and promoting the expressions of insulin signaling pathway proteins.
10. Effect of Canna edulis Type 3 Resistant Starch in Reducing Body Weight, Serum Lipid and Acute Toxicity
Min-yi QIU ; Xiao-yan JIN ; Cai-juan ZHANG ; Chi ZHANG ; Min WANG ; Yan-bing LU ; Bao-sheng ZHAO ; Yuan ZHANG ; Xue-yong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(1):149-156
Objective: To evaluate effect of Canna edulis type 3 resistant starch(RS3) on weight loss and lipid reduction in obese hyperlipidemia mice and acute toxicity in mice. Method: KKAy mice were fed with high-fat diet for 20 weeks to establish a hyperlipidemia model and then randomly divided into model group,positive group (4 mg·kg-1), high-dose resistant starch group and low-dose resistant starch group (2,1 g·kg-1).Mice in normal group were fed with standard diet. The medication groups received corresponding drugs by gavage. Normal group and high-fat model group were given equal volume of deionized water. After 8 weeks,mice were put to death. The levels of total cholesterol (TC),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were measured,and weigh fat mass,fat/body ratio,body fat rate and Lee's index were calculated accurately. The pathological changes of liver and adipose tissue were observed byhematoxylin-eosin (HE). The acute toxicity of RS3 to mice was evaluated by limit test. The mice were continuously observed for 14 days, and the toxicity of mice was recorded. Result: The indicators of high-dose RS3 group were significantly reduced,such as body weight,fat mass,body fat rate,fat/body ratio,Lee's index,and serum TC,TG,LDL-C,AST,ALT levels(P<0.05).Histomorphometric examination showed that the administration of RS3 starch could significantly improve the fatty lesions of liver tissue, and the liver-protecting effect was obvious, which could still inhibit the expansion of fat cells and reduce the accumulation of fat in mice.Among them, the high-dose group wsa better;After the maximum dose of 36 g·kg-1 was administered,no toxic reaction and death occurred in the animals. Conclusion: RS3-type Canna Edulis Resistant Starch has a good effect in reducing body weight and serum lipid,with a better effect in the high-dose group and no toxicity. And the commonly used clinical dose is safe and reliable.

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