1.Effects of Different Durations of Light Exposure on Body Weight and Learning and Memory Abilities of NIH Mice
Nan ZHANG ; Huaiyin LI ; Xiaodi LIAN ; Juanpeng WEI ; Ming GAO
Laboratory Animal and Comparative Medicine 2025;45(1):73-78
Objective This study aims to investigate the effects of varying durations of light exposure on body weight and learning and memory abilities of pubertal NIH mice. Methods Forty pubertal NIH mice, evenly split by gender and with similar initial weights, were subjected to a 12 h light-dark cycle for one week. They were then randomly assigned to groups with daily light exposure durations of 0, 6, 12, 18, and 24 hours, with 8 mice in each group. The experimental period lasted for 7 weeks, with the first 5 weeks as the feeding phase under different light exposure conditions, and the last 2 weeks as the behavioral testing phase. Their body weight was monitored, and learning and memory abilities were assessed using the T-maze, object location test, and eight-arm maze tests. Results During the light exposure period, there were no significant differences in body weight among groups (P>0.05). However, the weight gain of mice in the 24 h group was significantly higher than that of the 0 h group and the 6 h group during the second and third weeks of light exposure (P<0.05). After five weeks of light exposure, in the T-maze test, the latency time of the 0 h light exposure group was significantly longer than that of the 12 h group (P<0.01), and the latency time of the 24 h light exposure group was significantly longer than that of the 12 h group (P<0.05). In the object location test, the mice in 12 h group exhibited a higher discrimination index and spent more time observing the new location compared to the other groups, with significant differences in comparison to the 18 h group (P<0.01) and the 24 h group (P<0.05). In the eight-arm maze test, the time to find food, the reference memory error rate, and the working memory error rate in the 12 h group were all lower than those in the 0 h group, with significant differences (P<0.05). Moreover, the working memory error rate in the 24 h group was higher than that in the 12 h group, with significant differences (P<0.05). Conclusion Continuous 24 h light exposure affects body weight gain, while light exposure durations exceeding 18 h or below 6 h per day weaken the learning and memory abilities of NIH mice.
2.Exploration and Practice of Safe Access System Construction for Barrier Environment Facilities of Laboratory Animals: A Case Study on Xianlin Campus of Nanjing University
Dongxia HOU ; Zuoxiu TIE ; Yong LU ; Panpan NAN ; Jie BAO
Laboratory Animal and Comparative Medicine 2025;45(1):96-100
Laboratory animals are essential in scientific research and experimental teaching in colleges and universities. Disciplines such as life sciences, medicine, pharmacy, chemistry, and biomedical engineering heavily rely on animal experiments. The standardized barrier environmental facility for laboratory animals provides a fundamental platform for stable, scientific, and reliable animal experiment results. Rigorous access management for such facilities is a vital safeguard for maintaining standardized operations of facilities, controlling the quality and stability of laboratory animals, mitigating pathogen contamination risks among animals and laboratory staff, and preventing biosecurity incidents such as zoonotic disease outbreaks. Taking the small-scale barrier facilities for laboratory rats and mice at Nanjing University's Xianlin Campus, operational since 2019, as an example, this study focuses on the safety access management system of these facilities. Based on five years of operational data and accumulated experience in studying and optimizing the access management system, this study, from the perspectives of management system development and the formulation and implementation of standard operating procedures, reviews five aspects of access management: personnel access, animals access, material access, equipment access, and air circulation control. Furthermore, these aspects are systematically analyzed and summarized to serve as a reference for the construction and management of the laboratory animal facilities in universities, while also contributing to scientific research, public health security, and the well-being of experimental personnel.
3.Mechanism of postoperative abdominal adhesion formation and therapeutic prospect of mesenchymal stem cell exosomes
Xingzhou ZHANG ; Ming WEI ; Guoqiang DONG ; Wei DU ; Yiwen LUO ; Nan ZHANG
Chinese Journal of Tissue Engineering Research 2025;29(1):147-155
BACKGROUND:The formation of postoperative abdominal adhesions is a complicated process,and the prevention of postoperative adhesions is an urgent problem in clinic. OBJECTIVE:To analyze the mechanism of adhesion at cellular and molecular levels,and to provide theoretical basis for the prevention and treatment of adhesion by mesenchymal stem cell exosomes. METHODS:"Abdominal adhesion,pelvic adhesion,postoperative adhesion,epithelial mesenchymal transformation,mesenchymal stem cells,stem cell exosomes,mesenchymal stem cell exosomes"were selected as Chinese and English search terms.We searched PubMed,CNKI,and Chinese biomedical literature and screened relevant articles on postoperative abdominal adhesion and mesenchymal stem cell exosomal intervention published from inception to August 2023.After systematic analysis,54 articles were finally included for the review. RESULTS AND CONCLUSION:(1)Any pathological factors such as peritoneal inflammation,mechanical injury,tissue ischemia,and foreign body implantation cause peritoneal surface injury,resulting in postoperative abdominal adhesion.The formation process of adhesion includes the interaction of peritoneal mesothelial cell repair,inflammatory response,fibrinolytic system,coagulation pathway and other processes,involving a variety of cytokines and signaling pathways.Wnt/β-catenin pathway can induce fibrosis and angiogenesis,and cooperate with transforming growth factor-β/Smads signaling pathway to stimulate fibroblast proliferation and cause peritoneal fibrosis.Meanwhile,nuclear factor-κB signaling pathway up-regulates the expression of cellular inflammatory factors,promotes fibroblast proliferation,and plays a key role in the process of tissue fibrosis.(2)The paracrine function of stem cells is an important direction of molecular intervention in abdominal adhesions based on regenerative medicine.It can participate in a variety of complex cytokines and signaling pathways involved in abdominal adhesions.(3)Compared with traditional methods for treating abdominal adhesions,mesenchymal stem cell exosome has biological activity and is safe to use.Mesenchymal stem cell exosomes without special culture and expansion have lower immunogenicity,longer stability and other advantages,can guide a normal repair and healing through a variety of ways.(4)Mesenchymal stem cell exosome has been proven to be involved in regulating the above processes of adhesion formation in previous studies,showing potential application prospects in clinical studies.However,further clinical studies are needed to explore appropriate treatment options for mesenchymal stem cell exosomes to address the problem of clinical translation.
4.Improvement effects and mechanism of water extract from Chrysanthemum morifolium on muscle atrophy in rats after ischemic stroke
Ruocong YANG ; Hu QI ; Yuanlin GAO ; Zeyang ZHANG ; Xiaorui CHEN ; Rong LIU ; Nan ZENG
China Pharmacy 2025;36(5):535-539
OBJECTIVE To study the improvement effects and potential mechanisms of water extract from Chrysanthemum morifolium on skeletal muscle atrophy in rats after ischemic stroke. METHODS Sprague-Dawley rats were randomly divided into sham operation group, model group, ATP group (10 mg/kg), C. morifolium water extract high-dose and low-dose groups (1.08, 0.54 g/kg). Except for sham operation group, ischemic stroke models were induced in rats from the other groups using middle cerebral artery occlusion. Starting from the first day after surgery, rats in each group were given corresponding drug/normal saline intragastrically, once a day, for consecutive 7 days. On the 7th day post-surgery, the rats’ body weights were measured, and their motor functions were evaluated, including Longa scores, exercise distance, grip strength; the electrophysiological signals of the skeletal muscles in rats were measured; the pathological morphology of the soleus muscle in rats was observed; the levels of tumor necrosis factor-α (TNF-α) in serum and soleus muscle were measured; the expressions of proteins related to TNF-α/c-Jun N- terminal kinase (JNK)/mitogen-activated protein kinase (MAPK) signaling pathway in the soleus muscle were determined. RESULTS Compared with sham operation group, the body weight, grip strength and exercise distance of rats were decreased/ shortened significantly (P<0.01); additionally, there was a notable reduction in the interpeak value of skeletal muscle electrophysiology (P<0.05 or P<0.01). Longa score, as well as the levels of TNF-α in serum and soleus muscle, and the expression levels of TNF-α, phosphorylated JNK, phosphorylated MAPK, muscle ring-finger protein-1, and muscle atrophy Fbox- 1 protein in the soleus muscle, were all significantly elevated (P<0.01). The skeletal muscle cells of the soleus muscle in the model group showed significant atrophy, with a markedly decreased cross-sectional area (P<0.01). Compared with the model group, the levels of the aforementioned indicators were significantly reversed in C. morifolium water extract groups (P<0.05 or P< 0.01), and the skeletal muscle cells of the soleus muscle were markedly enlarged. CONCLUSIONS C. morifolium water extract can improve skeletal muscle atrophy in rats after ischemic stroke, the mechanism of which may be associated with suppressing the activation of the TNF- α/JNK/MAPK E-mail:19932015@cdutcm.edu.cn signaling pathway.
5.Progress in the application of poloxamer in new preparation technology
Xue QI ; Yi CHENG ; Nan LIU ; Zengming WANG ; Hui ZHANG ; Aiping ZHENG ; Dongzhou KANG
China Pharmacy 2025;36(5):630-635
Poloxamer, as a non-ionic surfactant, exhibits a unique triblock [polyethylene oxide-poly (propylene oxide)-polyethylene oxide] structure, which endows it with broad application potential in various fields, including solid dispersion technology, nanotechnology, gel technology, biologics, gene engineering and 3D printing. As a carrier, it enhances the solubility and bioavailability of poorly soluble drugs. In the field of nanotechnology, it serves as a stabilizer etc., enriching preparation methods. In gel technology, its self-assembly behavior and thermosensitive properties facilitate controlled drug release. In biologics, it improves targeting efficiency and reduces side effects. In gene engineering, it enhances delivery efficiency and expression levels. In 3D printing, it provides novel strategies for precise drug release control and the production of high-quality biological products. As a versatile material, poloxamer holds promising prospects in the pharmaceutical field.
6.Effect of Wenyang Huazhuo Formula (温阳化浊方) on Reproductive Aging,Ovarian Mechanical Micro-environment,and Offspring Reproductive Potential in Aged Model Mice
Jiaqi XU ; Xiaoli ZHAO ; Nan JIANG ; Kaixi LI ; Yafei DING ; Zimu WEN ; Yingying JIA ; Mengjun JIANG ; Tian XIA
Journal of Traditional Chinese Medicine 2025;66(6):612-620
ObjectiveTo explore the possible mechanisms of Wenyang Huazhuo Formula (温阳化浊方, WHF) in improving reproductive aging from the perspective of the ovarian mechanical microenvironment. MethodsThe experiment included five groups, 3-month group (20 female mice at 3 months of age), 6-month group (20 female mice at 6 months of age), 6-month + WHF group (20 female mice at 5 months of age treated with WHF), 9-month group (20 female mice at 9 months of age), and 9-month + WHF group (20 female mice at 8 months of age treated with WHF). The 6-month + WHF group and 9-month + WHF group were orally administered WHF 41.2 g/(kg·d) once daily for 4 consecutive weeks. The other three groups received no intervention. Reproductive hormone levels were measured by ELISA. HE staining was used to count the numbers of various stages of follicles. Ovarian hyaluronic acid (HA) content and collagen fiber content were measured to evaluate the ovarian mechanical microenvironment. Superovulation was performed to observe the number of eggs obtained, as well as the number of offspring and birth weight to assess fertility. The in vitro fertilization and blastocyst culture of oocytes from female offspring in each group were observed to evaluate the effect of WHF on offspring reproductive potential. ResultsCompared with the 3-month group, the 6-month group and 9-month group showed significantly decreased serum levels of gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), decreased ovarian collagen content, and reduced numbers of primordial and secondary follicles. In contrast, the numbers of primary follicles, antral follicles, and atretic follicles increased. The levels of anti-Müllerian hormone (AMH), ovarian HA content, and the fertilization rate, cleavage rate, and blastocyst formation rate of oocytes from offspring were significantly lower (P<0.05). Compared with the 6-month group, the 6-month + WHF group showed significantly reduced serum levels of GnRH, FSH, and LH, with a significant decrease in primary follicles, antral follicles, and atretic follicles as well as increase of AMH levels, ovarian HA content, number of primordial and secondary follicle, egg count, and offspring birth weight (P<0.05). Compared with the 9-month group, the 9-month + WHF group exhibited reduced GnRH, FSH, and collagen fiber content, as well as reduced number of primary follicles, antral follicles, and atretic follicles. However, AMH levels, ovarian HA content, number of primordial and secondary follicle, egg count, offspring numbers, birth weight, fertilization rate, cleavage rate, and blastocyst formation rate of oocytes from offspring all significantly increased (P<0.05). ConclusionWHF can significantly improve the ovarian reserve, fertility, and reproductive potential in offspring during reproductive mid-life and late-life stages. Its effect may be related to the remodeling of the mechanical microenvironment of aging ovaries. Moreover, the effect on the mechanical microenvironment remodeling of late-stage ovaries and the improvement of the offspring reproductive potential is more significant.
7.Antibacterial piezoelectric materials:no selective killing of bacteria and no bacterial resistance
Chinese Journal of Tissue Engineering Research 2025;29(10):2105-2112
BACKGROUND:Piezoelectric materials can catalyze the generation of reactive oxygen species,which can destroy bacteria by multiple ways without causing drug resistance.This indiscriminately attack bacteria strategy has obvious advantages over traditional antibiotic therapy,thus providing a novel idea for antibacterial strategies. OBJECTIVE:To summarize the properties and antibacterial mechanisms of piezoelectric materials and discuss the application status of several piezoelectric materials in the field of anti-bacteria. METHODS:The literature search was performed in PubMed,Web of Science,CNKI,and WanFang databases.Chinese search terms were"piezoelectric materials,piezoelectric catalysis,reactive oxygen species,antibacterial,bacterial infection,anti-infection,drug resistance."English search terms were"piezoelectric materials,piezoelectricity,piezoelectric catalysis,piezocatalysis,reactive oxygen species,ROS,bacterial infection,antibacterial strategies,anti-infection,drug resistance,drug-resistant bacteria."Retrieval time was from January 2013 to December 2023.Primary screening was conducted by reading the titles and abstracts.Repetitive studies and irrelevant articles were excluded.Finally,68 articles were included for review after literature quality evaluation. RESULTS AND CONCLUSION:(1)Piezoelectric materials are stable and environment-friendly materials,most of which show good biocompatibility.(2)Piezoelectric materials can catalyze a large amount of reactive oxygen species in the process of piezoelectric effect,combined with extracellular oxidation and intracellular oxidation,reactive oxygen species can destroy the membrane of bacteria,intracellular proteins,enzymes,and nucleic acids,disorder the structure and function,even kill the bacteria.The antibacterial performance is related to the rate of catalytic generation of reactive oxygen species,and the catalytic efficiency is related to many factors such as material system,morphology,and external conditions.(3)Reactive oxygen species producted by piezoelectric catalysis can kill bacteria without selectivity and show spectral antibacterial activity.This strategy does not rely on antibiotics and does not cause drug resistance.(4)Combined with the advantages of non-invasive,controllable,and penetrating ultrasound,piezoelectric materials will have significant value and great potential in the future as adjunctive or alternative treatments for drug-resistant bacterial infections and other fields.(5)The current challenge of low catalytic efficiency of piezoelectric materials limits its application in the field of antibacterial,how to improve the piezoelectric catalytic efficiency has become the focus of scholars'attention.
8.Material basis and action mechanism of drug-containing serum of Modified Erxian Pill inhibiting macrophage pyroptosis
Siyuan LI ; Yuru WANG ; Ye XU ; Di GUO ; Nan NAN ; Yang LIU ; Jie ZHAO ; Huiqin HAO
Chinese Journal of Tissue Engineering Research 2025;29(19):4029-4037
BACKGROUND:Our previous study found that Modified Erxian Pill could alleviate inflammation in collagen-induced arthritis rats,but its mechanism needs to be further verified. OBJECTIVE:To analyze the components absorbed in the blood of Modified Erxian Pill,and observe the effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages. METHODS:(1)Analysis of components absorbed in the blood of Modified Erxian Pill:Ultra-high performance liquid chromatography-high resolution mass spectrometry was used to detect and identify Modified Erxian Pill and its components absorbed in the blood.(2)Effect of the drug-containing serum of Modified Erxian Pill on pyroptosis of J774A.1 macrophages:Molecular docking technology was used to initially verify the sesquiterpenoids and NLRP3 in components absorbed in the blood of Modified Erxian Pill.J774A.1 macrophages were randomly divided into blank control group,lipopolysaccharide+adenosine triphosphate group,and lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill with low(2.5%),medium(5%),and high(10%)dose groups.The release of lactate dehydrogenase in the cell supernatant of each group was detected according to the kit instructions.The levels of interleukin-1β and interleukin-18 in cell supernatant were detected in each group by ELISA.The cell membrane damage was detected by Hoechst/PI staining.The expression levels of NLRP3,Caspase-1,GSDMD,and GSDMD-N protein in the cells of each group were detected by western blot assay. RESULTS AND CONCLUSION:(1)A total of 32 active components of Modified Erxian Pill were identified,and 21 components entered the blood.The main components into blood included a variety of sesquiterpenoids.(2)Molecular docking results showed that 3-O-Acetyl-13-deoxyphomenone,Incensol oxide,Atractylenolide III,Rupestonic acid,and 3,7-Dihydroxy-9,11-eremophiladien-8-one had good binding activity with NLRP3.(3)Compared with the blank control group,lactate dehydrogenase activity and the expression levels of interleukin-1β and interleukin-18 were significantly increased in cell supernatant of lipopolysaccharide+adenosine triphosphate group(P<0.001).Hoechst/PI staining showed that the number of PI-positive cells was significantly increased.After the intervention of lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group,all of them showed different degrees of reduction.(4)Compared with the blank control group,NLRP3,Caspase-1,GSDMD,and GSDMD-N protein expression levels were significantly increased in the lipopolysaccharide+adenosine triphosphate group(P<0.05).Compared with lipopolysaccharide+adenosine triphosphate group,the protein expressions of NLRP3,Caspase-1,GSDMD,and GSDMD-N were significantly decreased in the lipopolysaccharide+adenosine triphosphate+Modified Erxian Pill group(P<0.05),and had a certain dose dependence.These findings verify that the drug-containing serum of Modified Erxian Pill may inhibit the pyroptosis of J774A.1 macrophages by regulating the NLRP3/Caspase-1/GSDMD pathway.
9.UPLC-Q-TOF-MS Reveals Mechanisms of Modified Qing'e Formula in Delaying Skin Photoaging and Regulating Circadian Rhythm
Wanyu YANG ; Xiujun ZHANG ; Yan WANG ; Chunjing SONG ; Haoming MA ; Lifeng WANG ; Nan LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):88-97
ObjectiveTo reveal the active substances and mechanisms of modified Qing'e formula (MQEF) in delaying skin photoaging by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS),network pharmacology, and cell experiments. MethodsUPLC-Q-TOF-MS and a literature review were employed to analyze the transdermally absorbed components in mice after the topical application of MQEF. The potential targets of MQEF in treating skin photoaging were retrieved from databases.The compound-potential target network and protein-protein interaction network were constructed to screen the key components and core targets. A photoaging cell model was established by irradiating HaCaT cells with medium-wave ultraviolet B (UVB). The safe doses of bakuchiol (BAK) and salvianolic acid B (SAB) for treating HaCaT cells and the effects of BAK and SAB on the viability of cells exposed to UVB irradiation were determined by the cell counting kit-8 (CCK-8) method.The reactive oxygen species (ROS) fluorescent probe was used to measure the ROS production in the cells treated with BAK and SAB.The expression levels of genes related to oxidative stress,inflammation,collagen metabolism,and circadian rhythm clock were measured by Real-time PCR. ResultsA total of 24 transdermally absorbed components of MQEF were identified,which acted on 367 potential targets,and 417 targets related to skin photoaging were screened out,among which 47 common targets were predicted as the targets of MQEF in treating skin photoaging. MQEF exerted the anti-photoaging effect via key components such as BAK and SAB,which acted on core proteins such as serine/threonine kinase 1 (Akt1) and mitogen-activated protein kinase 3 (MAPK3) and intervened in core pathways such as the tumor necrosis factor (TNF) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling pathways.Compared with the model group,the administration of BAK and SAB increased the survival rate of HaCaT cells (P<0.01),down-regulated the mRNA levels of cyclooxygenase-2 (COX-2),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),matrix metalloproteinase-1 (MMP-1),and matrix metalloproteinase-9 (MMP-9) (P<0.01),and up-regulated the mRNA levels of heme oxygenase-1 (HO-1) and NAD(P)H quinone dehydrogenase 1 (NQO-1) (P<0.05,P<0.01) in photoaged HaCaT cells.In addition,it eliminated excess ROS production induced by UVB and up-regulated the mRNA levels of brain and muscle ARNT-like 1 (BMAL1) and circadian locomotor output cycles kaput (CLOCK) associated with circadian clock (P<0.05,P<0.01). ConclusionMQEF delays skin photoaging through the coordinated effects of various components,multiple targets,and diverse pathways.The key components BAK and SAB in MQEF exhibit anti-photoaging properties,which involve inhibiting oxidative stress,preventing collagen degradation,mitigating inflammation,and maintaining normal skin circadian rhythms by regulating clock gene expression.
10.Modified Ditan Tang Regulates Biorhythm-related Genes in Rat Model of Non-alcoholic Fatty Liver Disease
Zhiwen PANG ; Yu LIU ; Nan SONG ; Jie WANG ; Jingxuan ZHU ; Zhen HUA ; Yupeng PEI ; Qun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):115-124
ObjectiveTo investigate the effects of modified Ditan tang on genes related to the transcription-translation feedback loop (TTFL) of biorhythm in the rat model of non-alcoholic fatty liver disease (NAFLD) and its mechanism for prevention and treatment of NAFLD. MethodsSixty-five healthy SPF male SD rats were randomly assigned into blank (n=20), model (n=15), and low-, medium-, and high-dose (2.68, 5.36, and 10.72 g·kg-1·d-1, respectively) modified Ditan tang (n=10) groups. Other groups except the blank group were fed a high-fat diet for 12 weeks. The modified Ditan tang groups were treated with the decoction at corresponding doses by gavage, and the blank and model groups were treated with an equal volume of normal saline from the 9th week for 4 weeks. The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the serum were measured by an automatic biochemical analyzer. TG and non-esterified fatty acid (NEFA) assay kits were used to measure the levels of TG and NEFA in the liver. The pathological changes in the hypothalamus and liver were observed by hematoxylin-eosin staining, and the lipid deposition in the liver was observed by oil red O staining. The levels of brain-muscle ARNT-like protein 1 (BMAL1/ARNTL) in the hypothalamus and liver were determined by immunohistochemical staining. The mRNA and protein levels of BMAL1, circadian locomotor output cycles kaput (CLOCK), period circadian clock 2 (PER2), and cryptochrome1 (Cry1) in the hypothalamus and liver were determined by Real-time PCR and Western blot, respectively. ResultsCompared with the blank group, the model group showed elevated levels of TG, TC, LDL-C, AST, and ALT (P<0.01) and a lowered level of HDL-C (P<0.05) in the serum, elevated levels of TG and NEFA in the liver (P<0.01), pyknosis and deep staining of hypothalamic neuron cells, and a large number of vacuoles in the brain area. In addition, the model group showed lipid deposition in the liver, up-regulated mRNA and protein levels of CLOCK and BMAL1 (P<0.01), and down-regulated mRNA and protein levels of Cry1 and PER2 (P<0.01) in the hypothalamus and liver. Compared with the model group, all the three modified Ditan tang groups showed lowered levels of TG, TC, LDL-C, ALT, and AST (P<0.05, P<0.01) and an elevated level of HDL-C (P<0.05) in the serum, and lowered levels of TG and NEFA (P<0.05, P<0.01) in the liver. Furthermore, the three groups showed alleviated pyknosis and deep staining of hypothalamic neuron cells, reduced lipid deposition in the liver, down-regulated mRNA and protein levels of CLOCK and BMAL1 (P<0.05, P<0.01), and up-regulated mRNA and protein levels of Cry1 and PER2 (P<0.05, P<0.01) in the hypothalamus and liver. ConclusionModified Ditan tang can reduce lipid deposition in the liver and regulate the expression of CLOCK, BMAL1, Cry1, and PER2 in the TTFL of NAFLD rats.

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