Abstract: Objective　To investigate the regulatory role of nuclear factor erythroid 2-related factor 2 (Nrf2) in the activation and polarization of microglia induced by methamphetamine (METH). Methods　BV2 and HMC3 cells were studied in vitro and wild-type mice and Nrf2-knockout mice were studied in vivo. In vivo and in vitro toxicity models induced by METH were established, respectively. The activation and polarization of microglia in each group were examined using immunofluorescence and Western blot, respectively. Results　METH treatment significantly increased the fluorescence level of inducible nitric oxide synthase (iNOS) in BV2 and HMC3 cells (P<0.001), and significantly decreased the fluorescence level of Arginase1 (Arg1) (P<0.05, P<0.01). METH exposure activated microglia in the cortex, increased expression levels of ionized calcium binding adaptor molecule-1 (IBA1) and iNOS (P<0.001, P<0.05), and decreased the expression of Arg1 (P<0.01). The number of activated microglia was significantly increased after Nrf2 gene knockout (P<0.01), compared with the WT METH group, while the expression levels of IBA1 and iNOS were also increased (P<0.001, P<0.01) and the expression level of Arg1 was decreased (P<0.01). Conclusions　Nrf2 plays an important role in regulating the activation and polarization of microglia induced by METH. Nrf2 may thus be a potential target for the treatment of neuroinflammation induced by METH.

Abstract: Objective　To observe the influence of Creb protein over-expression and under-expression in the prefrontal cortex on depressive behavior in rats. Methods　Adeno-associated virus (AAV) strains that can knock down Creb expression in the prefrontal cortex of rats were injected using the stereotaxic injection method and screened by Western blot, reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence. Forty rats were divided randomly into Control, chronic restraint stress (CRS), CRS combined with AAV interference (CRS+AAVI), and CRS combined with AAV overexpression (CRS+AAVO) groups. The body weight and food intake of the rats in each group were monitored during establishment of the animal model. After establishment of the model, behavioral changes in the rats were monitored by sucrose preference, elevated plus maze, forced swimming, and open field tests. The 5 hydroxytryptamine (5-HT), norepinephrine (NE), and corticosterone (CORT) contents in the prefrontal cortex of rats in each group were measured by enzyme-linked immunosorbent assay. Results　Western blot and immunofluorescence showed that Creb protein expression was significantly reduced in the short hairpin RNA2 (shRNA2) knockdown groups compared with the other groups (P<0.05). RT-qPCR showed that Creb mRNA expression was also significantly reduced compared with the other three groups (P<0.01). The AAV-CREB1-shRNA2 virus strain was therefore selected for subsequent Creb knockdown experiments in this study. After modeling, the food intake of rats in the CRS+AAVI group was significantly reduced compared with the other groups (P<0.01). Rats in this group also showed slow weight gain and decreased desire to explore new environments, significantly increased despair and nervous behavior, and significantly decreased 5-HT and NE levels(P<0.01) and significantly increased CORT levels in the prefrontal cortex (P<0.01). These depressive behaviors and associated neurotransmitter levels were reversed in the CRS+AAVO group. Conclusions　Lower expression of Creb in the prefrontal cortex can aggravate the degree of depression in rats, while high expression of Creb can alleviate depression to a certain extent. These result confirm that Creb expression in the prefrontal cortex is an important target in the pathogenesis of depression, thus providing ideas and references for the construction of animal gene models and further studies of the pathogenesis of depression.

Abstract: Objective　To investigate the neuroprotective effect and mechanism of glycosides of Cistanche (GCs) on cerebral ischemia reperfusion injury (CIRI) in rats. Methods　Forty-eight male Wistar rats were divided randomly into Sham, Model, GCs, and Nim groups. A rat model of focal CIRI was established by middle cerebral artery occlusion. Neurological function was scored using the Zea-Longa scoring method. The sensory and motor abilities of rats in each group were evaluated by sticker removal, balance beam, and open field tests. The area of cerebral infarction was detected by 2,3,5-triphenyltetrazolium chloride(TTC) staining, Nissl staining was used to observe the morphology of nerve cells, and terminal deoxynucleotidyl transferase dUTP nick end labeling was used to detect apoptosis of nerve cells. Expression levels of the apoptosis-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), and cysteine aspartic protease-3 (Caspase-3) were detected by immunohistochemical staining and Western blot. Results　Compared with the Sham group, the neurological deficit score was significantly increased (P<0.05) and the times to remove stickers and passing the balance beam were significantly increased (P<0.05), motor ability was decreased, infarct size was increased, the number of neurons was decreased, and the number of apoptotic cells was increased after CIRI. Bax and Caspase-3 expression were significantly increased (P<0.05) and Bcl-2/Bax was significantly decreased (P<0.05). Compared with the Model group, GCs improved the behavioral performance of CIRI model rats, reduced the infarct size, inhibited cell apoptosis, downregulated the expression of Bax and Caspase-3 (P<0.05), and up-regulated the expression of Bcl-2/Bax (P<0.05). Conclusions　GCs have a neuroprotective effect on CIRI, and may play a role in inhibiting cell apoptosis by regulating the expression of the apoptosis-related factors Bax, Bcl-2, and Caspase-3.

Abstract: Objective　To investigate the mechanism by which Xuanfei Jiedu Formula (XFJDF) ameliorates pulmonary damage in rats with multidrug-resistant Pseudomonas aeruginosa (MDR-PA) pneumonia, by modulating the activity of the inhibitor of nuclear factor (NF)-κB kinase (IKK) IKK/NF-κB signal transduction cascade. Methods Eighty-four rats were divided randomly into seven groups: control, model, XFJDF-low dose, XFJDF-medium dose, XFJDF high dose, imipenem (IPM), and pyrrolidinedithiocarbamate ammonium (PDTC) groups (n=12 rats per group). A MDR-PA pneumonia rat model was established by oral tracheal intubation. After successful model construction, rats in the low-, medium-, and high-dose XFJDF groups were given the corresponding dose of drugs by gavage, rats in the IPM group were given an intraperitoneal injection of IPM, and rats in the control and model groups were given the same volume of normal saline by gavage, twice a day for 7 days. PDTC was injected intraperitoneally 1 h before model establishment, 12 h and 24 h after model establishment in the NF-κB inhibitor group. The behavior status, body weight changes, spleen and thymus indexes, and lung wet weight/dry weight ratio were observed in the different groups. Histological changes in the lung tissue were assessed by hematoxylin and eosin staining. Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to detect apoptosis of lung tissue cells, and serum levels of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β, and IL-10 were detected by enzyme-linked immunosorbent assay, and glutathione (GSH) and malondialdehyde (MDA) levels, myeloperoxidase (MPO) activity and total antioxidant capacity (T-AOC) were determined by colorimetry and thiobarbituric acid assay. NF-κBp65 in lung tissue was identified by immunohistochemical analysis, IKKβ and NF-κBp65 mRNA were analyzed by reverse transcription quantitative polymerase chain reaction, and expression levels of IKKβ, phosphorylated (p)-IKKβ, NF-κBp65, and p-NF-κBp65 were measured by Western blot. Results　Compared with the control group, rats in the model group exhibited decreased appetite, dull fur, sluggish responsiveness, decreased mobility, increased respiratory rate, audible murmurs, and notable weight loss (P<0.01). The spleen and thymus indices were significantly enhanced (P<0.01) and the lung wet/dry weight ratio was significantly increased (P<0.01), concurrent with increased alveolar secretions in lung tissue. Infiltration of abundant inflammatory cells and increased apoptosis in lung tissue were also noted. Serum concentrations of IL-1β, TNF-α, TGF-β, and IL-10 were increased (P<0.01) and the MDA content and MPO activity were also increased (P<0.01). Conversely, GSH levels and T-AOC were significantly decreased (P<0.01). Lung levels of IKKβ and NF-κBp65 mRNA were significantly increased (P<0.01) and the ratios of p-IKKβ/IKKβ and p-NF-κBp65/NF-κBp65 were also significantly increased (P<0.01) compared with the control group. XFJDF, IPM and PDTC improved these parameters to varying degrees, compared with the model group (P<0.05, P<0.01), with particularly significant effects in the high-dose XFJDF and IPM groups. Conclusions　XFJDF may improve lung injury in rats with MDR-PA pneumonia by inhibiting the IKK/ NF-κB signaling pathway.

Abstract: Objective　miR-207 is differentially expressed in many diseases. We investigated the mechanism by which miR-207 overexpression regulates the survival of Mycobacterium tuberculosis (H37Ra) in macrophages, to provide a theoretical basis for the targeted therapy of tuberculosis. Methods　Macrophages were divided into four groups: blank (Ana-1 cells), control (cells infected with H37Ra), mi (infected with H37Ra and transfected with miRNA-207 mimics), and mi-NC (infected with H37Ra and transfected with NC mimics) groups. A model of tuberculosis infection was established using H37Ra-infected Ana-1 cells, and miRNA-207 and NC mimics were transfected into Ana-1 cells using the liposome transfection method. Tuberculosis colony-forming units were counted to assess the effect of miR-207 on intracellular mycobacterial load and clearance of extracellular residual mycobacteria. The total apoptosis rate was detected by flow cytometry. The relative expression levels of miR-207 and apoptosis, pyroptosis, inflammation, and autophagy genes were measured by quantitative real-time polymerase chain reaction (qPCR). Relative expression levels of apoptosis, pyroptosis, and autophagy proteins were detected by Western blot. Fluorescence microscopy and multifunctional enzyme labeling were used to detect the fluorescence intensity of intracellular reactive oxygen species (ROS) and lactate dehydrogenase (LDH). Results　Successful establishment of the infection model was observed under the microscope. qPCR showed that miR-207 expression was lower in the control compared with the blank group(P<0.01), indicating differential expression between these two groups. miR-207 expression was significantly higher in the mi compared with the mi-NC group(P<0.0001), indicating successful establishment of the transfection model. The number of colonies and total apoptosis were both higher in the mi group compared with the mi-NC and control groups(P<0.001). qPCR and Western blot showed that the relative expression levels of apoptotic genes and proteins were higher in the control group than in the blank group(P<0.05), and higher in the mi group than in the mi-NC group(P<0.05). The relative expression levels of inflammatory genes were higher in the control than in the blank group(P<0.001). The relative expression levels of inflammatory genes were higher in the mi group than in the mi-NC group(P<0.05), and the relative expression levels of pyroptosis genes and proteins were higher in the control group compared with the blank group(P<0.01) and higher in the mi group compared with the mi-NC group(P<0.05). The relative expression levels of the autophagy positively-regulated genes LC3 and Beclin1 were higher in the control compared with the blank group(P<0.0001), and lower in the mi than in the mi-NC group(P<0.05), while negatively-regulated autophagy genes showed the opposite trend. Autophagy-related proteins showed similar trends to the autophagy genes. ROS fluorescence intensity was higher in the control compared with the blank group(P<0.05), and higher in the mi compared with the mi-NC group(P<0.001). LDH content was higher in the control than in the blank group(P<0.01), but there was no significant difference between the mi and mi-NC groups(P>0.05). Conclusions　miR-207 overexpression promotes apoptosis, cellular pyroptosis, and inflammation, inhibits autophagy, and favors H37Ra survival. These result provide a potential new direction for the treatment of tuberculosis.

Abstract: Objective　To compare the reparative effects of moxibustion at “Guanyuan” and “Shenshu” points in rats with a thin endometrium. Methods　Thirty-two female Sprague-Dawley rats were divided randomly into control(CON), model(MOD), Guanyuan(GY), and Shenshu(SS) groups. A thin endometrium model was established using 95% anhydrous alcohol. The Guanyuan and Shenshu groups underwent moxibustion at the Guanyuan and Shenshu points, respectively, and the other two groups of rats were fixed in the same way. Temperature changes at the acupoint area were measured at different time points. The pathological morphology of the uterus was observed by hematoxylin and eosin (HE) staining, and serum levels of inflammatory factors and hormone levels were detected by enzyme-linked immunosorbent assay (ELISA). Expression levels of proliferation and endometrial receptivity factors were detected by immunohistochemistry (IHC), and angiogenesis factors were detected by Western blot. Results　The temperatures immediately after moxibustion and after 5 min were higher at the Shenshu compared with the Guanyuan acupoint (P<0.01). Endometrial thickness and numbers of glands and blood vessels were decreased in the MOD group compared with the CON group (P<0.01), serum interleukin 1β (IL-1β) was increased (P<0.01) and PROG was decreased (P<0.01), and Ki67, proliferating cell nuclear antigen (PCNA), leukemia inhibitory factor (LIF), integrin β3 (ITG-β3),angiopoietin-1 (ANG-1), and CD34 protein in uterine tissue were all decreased (P<0.01). Endometrial thickness and numbers of glands and blood vessels were increased in the GY and SS groups compared with the MOD group (P<0.01), while serum IL-1β was decreased (P<0.05), E2 and PROG were increased (P<0.05), and serum interleukin 10 (IL-10) was increased in the SS group (P<0.05). Ki67, PCNA, LIF, and ITG-β3 were all increased in both treatment groups compared with the MOD group (P<0.01). PCNA was higher in the GY group compared with the SS group (P<0.05), while LIF and ITG-β3 were higher in the SS group compared with the GY group (P<0.01, P<0.05, respectively). CD34 and VEGFA levels were increased in the two treatment groups compared with the model group (P<0.01). ANG-1 was higher in the GY group than that in the SS group (P<0.01), while CD34 was higher in the SS group than that in the GY group (P<0.05). Conclusions Moxibustion at the Guanyuan and Shenshu points could help to repair the pathological thin endometrium in rats. The GY group may be superior to the SS group for reducing the levels of serum IL-1β , increasing E2, and promoting the expression of proliferation factors, while the SS group may be superior to the GY group for increasing the levels of PROG and promoting the expression of endometrium receptor-related factors. Both acupoints can promote the expression of angiogenesis factors, potentially acting via different pathways.

Abstract: Objective　We aimed to investigate the effect of roofplate-specific spondin 2 (RSPO2) on oxidative stress and apoptosis in ovarian granulosa cells, using human ovarian granulosa tumor cells as a cellular model. Methods We transiently transfected COV434 human granulosa cells with RSPO2 overexpression plasmid and small interfering RNA (siRNA). The efficiencies of the overexpression and interference were detected by quantitative real-time polymerase chain reaction (qPCR) and Western blot. Intracellular reactive oxygen species (ROS) were detected using a ROS assay kit, and apoptosis was detected by flow cytometry. Expression levels of oxidative stress- and apoptosis-related genes were determined by qPCR and Western blot, and interacting proteins were detected by co-immunoprecipitation. Results　RSPO2 overexpression and interference were successfully realized. Overexpression of RSPO2 significantly inhibited ROS levels in COV434 cells (P<0.05) and the early apoptosis of granulosa cells (P<0.01), while RSPO2 interference had the opposite result. Overexpression of RSPO2 significantly upregulated the mRNA and protein levels of superoxide dismutase (SOD) 1, SOD2, and catalase (CAT) (P<0.05), and significantly downregulated the levels of Caspase 3 and Caspase 8 (P<0.05). In contrast, RSPO2 interference significantly downregulated SOD1 and CAT (P<0.05) and significantly upregulated Caspase 3 and Caspase 8 (P<0.05). We found protein interactions between RSPO2, SOD1, CAT, and Caspase 3. Conclusions　Interfering with RSPO2 promotes the accumulation of ROS within cells and downregulates the expression of SOD1 and CAT, thereby enhancing oxidative stress. Additionally, interfering with RSPO2 upregulates the expression of Caspase 3 and Caspase 8, promoting cell apoptosis. These findings suggest that RSPO2 plays a crucial role in ovarian granulosa cells.

Abstract: Objective　To prepare human SET8 monoclonal antibody and explore its effects on the proliferation, apoptosis, and cell cycle of hepatoma cells, and to evaluate its anti-tumor effect in mouse models of hepatocellular carcinoma. Methods　We immunized mice with human SET8 polypeptide fragment and screened and fused B cells and myeloma cells to establish a hybridoma cell line that stably secreted SET8 monoclonal antibody. Production was expanded by intraperitoneal injection into mice and the collection and purification of ascites. We investigated the effects of SET8 monoclonal antibody on the proliferation, apoptosis, cell cycle, and apoptosis-related protein expression of hepatocellular carcinoma cells by CCK-8, flow cytometry, and Western blot, respectively. Finally, we constructed a mouse model of human hepatocellular carcinoma by cell transplantation to evaluate the inhibitory effect of SET8 monoclonal antibody on tumor growth in vivo. Results　Human SET8 monoclonal antibody significantly inhibited the viability of Huh-7 and Mahlavu hepatoma cells at concentrations of 50 and 100 μg/mL, in a concentration-dependent manner (P<0.05). Flow cytometry analysis showed that SET8 monoclonal antibody, paclitaxel, and their combination significantly increased the apoptosis rate of Mahlavu cells compared with the blank control group, with the combination group having the greatest effect (P<0.05). SET8 monoclonal antibody also induced Mahlavu cell cycle arrest in S and G₂ phases and reduced G₁ phase cells. Western blot analysis showed that the monoclonal antibody increased the expression of the apoptosis-related proteins Bax and Caspase-3 (P<0.05). SET8 monoclonal antibody, alone or in combination with paclitaxel, also effectively inhibited the proliferation of hepatocellular carcinoma cells in nude mice, with the combination therapy having the most significant effect (P<0.05). Conclusions　The prepared human SET8 monoclonal antibody effectively inhibited the proliferation and promoted the apoptosis of hepatocellular carcinoma cells, and showed good anti-tumor effects in mice.

Abstract:The elective course “Practical Mouse Anatomy & Comparative Medicine” is a comprehensive technical course designed to cultivate fundamental theoretical knowledge and experimental skills in laboratory mouse anatomy for undergraduate students in medical and pharmaceutical-related fields. Given the increasing demand for innovation and entrepreneurship training programs and undergraduate participation in research laboratories, this course systematically provides students with anatomical knowledge of the eight major systems and local anatomy of experimental mice, emphasizing the combination of theory and practice, to provide a comprehensive learning platform. The course continually explores and reforms its practices to ensure that the teaching content keeps pace with the forefront of scientific research, fostering a rigorous and pragmatic scientific attitude in students, thereby laying a solid foundation for their future research work. This paper discusses the effectiveness of the course in enhancing students’ research literacy and practical abilities, aiming to provide a reference for the design and implementation of similar courses.

Abstract:Insomnia is a prevalent clinical condition that not only diminishes the patient’s quality of life but also has the potential to give rise to further health complications. Traditional Chinese medicine (TCM) has been used to treat insomnia for thousands of years, with distinct advantages. TCM uses syndrome differentiation and therapy to achieve its therapeutic effects, by regulating the internal balance of the human body. To gain a deeper understanding of how TCM treats insomnia, it is crucial to create animal models that exhibit insomnia symptoms closely resembling those found in humans. This review summarizes the current disease-syndrome combination models, which can be classified into four categories: liver depression and Qi stagnation, heart-spleen deficiency, heart-kidney incompatibility, and Yin and blood deficiency. This paper focuses on specific modelling method ologies, assessment indicators, and model performances, with the aim of enhancing our understanding of the pathophysiology of insomnia in the context of TCM, and providing technical assistance for clinical efficacy evaluation and the creation of novel drugs.

Abstract:Parkinson’s disease(PD) is a neurodegenerative disease that primarily affects middle-aged and older adults. Its main clinical manifestations include resting tremor, bradykinesia, myotonia, and balance disorders. Berberine is a naturally occurring isoquinoline alkaloid. Several studies over the previous decade have reported that berberine plays a crucial role in the pathophysiologic processes of a range of diseases, including playing an active role in the development of PD via different pathways. This paper systematically reviews the mechanism of action of berberine in the prevention and treatment of PD and the current status of research, to provide references for the future clinical application of berberine for the treatment of PD.

Abstract:The pathogenesis of depression is complex and treatment method are currently limited, and it is therefore important to clarify its pathogenesis and develop new therapeutic drugs. Hormones and hormone-modifying compounds have been used to treat depression, and their therapeutic effects are attributed to regulation of the peripheral endocrine system and their effects on non-endocrine circuits in the central nervous system. The hypothalamic-pituitary adrenal (HPA)/hypothalamic-pituitary-thyroid (HPT)/hypothalamic-pituitary-gonadal (HPG) axes are closely related to the pathogenesis of depression. The mechanism of action of traditional Chinese medicines in the treatment of depression is extensive, and the three axes are regulated in a variety of ways. Based on new evidence of hormones and hormone operon compounds involved in the pathology and treatment of depression, this review summarizes the relevant mechanisms of current traditional Chinese medicine in the treatment of depression, to provide new ideas and references for traditional Chinese medicine research.

Abstract:Hypoxic/ischemic preconditioning(H/IPC) can induce endogenous protective mechanisms that increase the tolerance of nerve cells to hypoxia/ischemia. This protective mechanism involves changes in gene expression during the critical decision-making period between cell survival and death. DNA methylation, as a crucial mechanism for gene expression regulation, plays an essential role in hypoxia/ischemia tolerance. DNA methyltransferases (DNMTs) contribute to neuroprotection by influencing gene expression via regulating DNA methylation levels. DNMTs thus have important functions in the neuroprotection induced by hypoxic/ischemic preconditioning. This paper reviews the role of DNMTs in this process, providing insights into neuroprotection targeting DNMTs.

Abstract:Methamphetamine(METH) is a new type of abused drug with a strong central stimulant effect. Its excitatory pathway involves a series of central higher-level functions, including drug-related rewards and motivation, learning and memory, decision-making, and execution. METH not only causes neurotoxicity, but its long-term use may also cause cognitive dysfunction, loss of personality, and may lead to serious social violence in addicts. Exosomes, as a novel cell-to-cell carrier, are involved in the occurrence and development of drug addiction, and exosome microRNAs are important biomarkers for METH addiction and are also involved in various aspects of METH-induced neurotoxicity. Eoxosmes may thus be useful therapeutic carriers providing a new approach for the diagnosis and treatment of METH abuse.

Abstract:The incidence of diarrhea-predominant irritable bowel syndrome (IBS-D) remains high, with microRNA (miRNA)-mediated intestinal barrier dysfunction, visceral hypersensitivity, low-grade inflammation, and dysbiosis playing significant roles in its pathogenesis. Traditional Chinese medicines can treat IBS-D by directly or indirectly targeting miRNAs to regulate multiple pathways and targets in a coordinated manner. This article systematically reviews the involvement of miRNAs in the pathogenesis of IBS-D and the progress of traditional Chinese medicine interventions. It explores the relationship between traditional genetics, epigenetics, and the Chinese medicine concepts of ‘kidney’ and ‘spleen’ in terms of congenital and acquired interactions from a molecular biology perspective. This review thus provides a reference for exploring the micro-material basis of the ‘Zang-Xiang’ theory of traditional Chinese medicine, and offers new ideas and method for the effective treatment of IaBS-D using traditional Chinese medicine.

Abstract:Spinal cord injury (SCI) is a central nervous system disease with high morbidity, disability, and mortality. A series of pathological and physiological changes after SCI, including oxidative stress, can promote further deterioration of the microenvironment at the injury site, resulting in impaired neurological function. Nuclear factor E2related factor 2 (Nrf2) is highly correlated with oxidative stress, suggesting that targeting the regulation of Nrf2 and alleviating oxidative stress may be an effective treatment for SCI. We consider the application of Nrf2 in post-SCI oxidative stress, based on the occurrence of oxidative stress after SCI and the relationship between oxidative stress and Nrf2. We also summarize the strategies for targeting and regulating Nrf2, including genes, non-coding RNAs, and drugs, with the aim of providing new ideas for targeted therapy of SCI.

Abstract:Autophagy is a crucial physiological process in cells and an important mechanism in the maintenance of cell homeostasis and regulation of cell survival. Idiopathic pulmonary fibrosis (IPF) is an interstitial lung disease of unknown etiology with a poor prognosis, characterized by alveolar epithelial cell damage and abnormal proliferation and activation of fibroblasts. IPF is accompanied by the excessive deposition of extracellular matrix(ECM), and its pathogenesis involves a complex interaction between cell types and signaling pathways. Defects in autophagy function have been shown to play a key role in the occurrence and development of IPF. This review aims to investigate the mechanism of autophagy in IPF and reveal its complexity and related signaling pathways.