Abstract: Objective To explore the dynamic changes in monoamine oxidase A (MAOA) and forkhead box A1 ( FOXA1) levels during neuroendocrine differentiation (NED) in prostate cancer, providing new strategies for the treatment of neuroendocrine prostate cancer. Methods Cell models and mouse transplantation models of NED were established through long-term sustained induction with enzalutamide(ENZ). Dynamic expression of MAOA and FOXA1 in NED was detected by Western Blot and Real-time PCR. GEO database data were selected to analyze the dynamic trends in MAOA and FOXA1 levels in multiple NED models. We constructed a mouse transplantation model of human prostate cancer cell lines and analyzed the dynamic expression of MAOA and FOXA1 in the in vivo NED model by immunohistochemistry. MAOA expression was disrupted with lentiviral transfection, and the impact on FOXA1 was detected. Results Both MAOA and FOXA1 concentrations showed dynamic characteristics, increasing and then decreasing during the NED process. Knockdown of MAOA in prostate cancer cells led to decreased expression of FOXA1. This MAOA may play different roles at different stages of NED by acting through FOXA1. Conclusions Both MAOA and FOXA1 levels showed increasing, then decreasing, trends during NED. The expression of MAOA affected the level of FOXA1, and MAOA/FOXA1 may play a dynamic regulatory role in the NED process.

Abstract: Objective To establish interferon-induced transmembrane protein 3 ( Ifitm3) knockout mice and to explore the effects of Ifitm3 on the proliferation and differentiation of adult neural stem cells of mice ( aNSCs). Methods IFITM3 knockout mice were established by the CRISPR/ Cas9 method and identified by genotype identification and Western Blot. The differences between Ifitm3-knockout mice and wild-type mice were analyzed by hematoxylin-eosin(HE) staining and flow cytometry. The aNSCs of wild-type mice and Ifitm3-knockout mice were isolated and cultured, the number and size of neurospheres were detected, The ability of aNSCs to proliferate and differentiate were detected by quantitative reverse-transcription polymerase chain reaction, Western Blot, and immunofluorescence. Results Ifitm3-knockout mice were successfully established. The mice developed normally, and there were no obvious abnormalities either histopathologically or the immune system. In vitro experiments showed that Ifitm3 knockout inhibited the self-renewal potential of aNSCs, led to a decrease in the proliferation ability of aNSCs, and inhibited the differentiation of aNSCs into immature neurons and astrocytes. Conclusions This study finds that a lack of IFITM3 result in the ability of aNSCs to proliferate and differentiate decreased, IFITM3 may regulate the function of aNSCs.

Abstract: Objective This study was performed to develop and assess a genetically engineered mouse model for visualizing in vivo fluorescence of glioma cells, mural cells, and blood vessels using two-photon microscopy. Methods PDGFRβ-Cre^{+ / -}:Rosa26-tdTomato^{+ / -}genetically engineered mice underwent skull clearance and were injectedwith GL261-CFP. This was performed to study the dynamic alterations in blood vessels and mural cells during the progression and invasion of glioma using two-photon microscopy. Results PDGFRβ-Cre^{+ / -}:Rosa26-tdTomato ^{+ / -} mice were successfully bred and subjected to hematoxylin-eosin section analysis of functional organ tissues. The mice exhibited no discernible differences from C57BL/ 6 mice in terms of appearance and morphology. Cre recombinase activity was fully induced following tamoxifen treatment on day 7. Subsequent GL261-CFP inoculation demonstrated the dynamic progression of glioma proliferation and invasion, as well as vascular abnormalities and increased mural cell detachment within the tumor. Conclusions Genetically engineered mice expressing fluorescent mural cells were successfully bred. Blood vessels labeled with fluorescein isothiocyanate-dextran and blue fluorescent tumor cells were utilized. Glass discs and fixed rings were employed to replace the skulls of the mice. This allowed for the tracking of morphological and structural changes in blood vessels and vascular supporting cells following the development of brain tumors in vivo over an extended period. This model offers a valuable tool for studying brain diseases through pathological visualization.

Abstract: Objective To assess the sweat secretion of Yin-deficient ovariectomized rats and investigate the changes in biochemical indexes. Methods Eighteen SD female rats were randomly divided into a sham operation group,model group, and positive control group of six rats each. The rats in the sham operation group underwent a sham operation,and those in the model group and positive control group underwent bilateral ovariectomy. L-Thyroxine ( 92 mg / kg) was given once a day for 7 consecutive days starting on the 7th postoperative day to establish a Yin-deficient ovariectomized model. The rats in the positive control group were orally administered Qinggu San Tang ( 7. 3 g / kg) once a day, while those in the sham operation group and model group were orally administered an equal amount of distilled water once a day for a total of 14 days. Sweat secretion from the plantar region of the foot was measured using the Wada-Takagaki reagent coloring method. At the end of the experiment, blood was taken from the abdominal aorta and the tissue of the paw pads was separated. The serum levels cyclic adenosine monophosphate ( cAMP), cyclic guanosine monophosphate ( cGMP),luteinizing hormone ( LH), gonadotropin-releasing hormone ( GnRH), and estradiol ( E₂ ) were measured by enzymelinked immunosorbent assay. Western Blot was used to determine the expression levels of M₃R, β₂AR, and aquaporin-5(AQP₅ ) in the paw pad. Results The three main findings of this study were as follows. (1) Compared with the rats in the sham operation group, those in the model group were more irritable and aggressive, and their body weights decreased while their average temperature and sweat secretion significantly increased. (2) Serum cAMP level and cAMP / cGMP ratio increased, the LH and GnRH levels significantly increased, and the E₂ level decreased. (3) M₃R expression was downregulated and β₂AR and AQP₅ expression was up-regulated in the paw pads of the rats. After 2 weeks of positive control treatment, the serum cAMP level and cAMP / cGMP ratio significantly decreased and the LH and GnRH levels decreased;however, no statistically significant difference was observed in the serum E₂ level. The expression levels of M₃R were increased-regulated in paw pads of the rats, and reduced expression of β₂AR and AQP₅ . Conclusions Sweat secretion significantly increased in this “combined disease and evidence” model of perimenopausal syndrome kidney yin deficiency established by desiccation combined with thyroxine. The underlying mechanism may be related to the changes in cGMP,cAMP, and key proteins M₃R, β₂AR, and AQP₅ in sweat glands that regulate sweat secretion.

Abstract: Objective Study on mechanism of Xuanfei Jiedu Formula in treating multi-drug resistant Pseudomonas aeruginosa pneumonia. Methods Except the Control group, the MDR-PA (9 × 10⁸ CFU/ mL, 0. 5 mL) pneumonia rat model was established by tracheal intubation, and an un-modeled control group was used. After successful modeling, rats were randomly divided into model group, XFJDF-low dose group, XFJDF-medium dose group, XFJDF-high dose group and IPM group, with 12 animals in each group; In addition to the blank group and the model group, the remaining drug administration groups were collectively referred to as the intervention treatment group. One day after modeling, the XFJDFlow dose, XFJDF-medium dose, and XFJDF-high dose groups were given the corresponding doses by gavage. The imipenem and cilastatin group was given Imipenem intraperitoneal injection, and the control and model groups were given saline gavage twice a day for 7 days. The rats’ general status, body weight changes, and wet-dry weight ratio (W/ D) of the lung tissue were observed. Hematoxylin-eosin staining was used to observe the pathological changes to the lung tissue of rats in each group under a light microscope. The serum levels of IL-1β, TGF-β, TNF-α, and IL-6 were detected by enzyme immunosorbent assay. Serum GSH content and MPO activity of rats were detected by colorimetry. The serum content of MDA was detected by TBA method. The total antioxidant capacity (T-AOC) was detected by kit method. The protein levels of TLR4, Myd88, and NF-κBp65 in lung tissues were determined by immunohistochemistry. The mRNA and protein levels of TLR4, Myd88, and NF-κBp65 in the lung tissues of each group were detected by qPCR and Western Blot. Results Compared with the control group, the model groups had delayed responses, increased respiratory frequency,increased respiratory noise, and appeared to have different degrees of chills, as well as decreased diet and water intake and decreased body weight. The W/ D of lung tissue was significantly increased ( P<0. 01 ), and a large number of inflammatory cells had infiltrated the alveolar cavity and around the lung bronchus. Some alveolar walls were fractured and fused to form air cavities, with inflammatory exudation, pulmonary interstitial thickening, and local lung fiber formation.The serum levels of IL-1β, TNF-α, TGF-β, and IL-6 were significantly increased ( P<0. 01), MDA content was increased, MPO activity was enhanced, GSH content and T-AOC capacity were decreased (P<0. 01), and the mRNA and protein levels of TLR4, Myd88, and NF-κBp65 in lung tissue were significantly increased (P<0. 01). Compared with the model groups, the intervention group and the treatment group showed improvements in the above indexes (P<0. 05, P<0. 01), and the Xuanfei Jiedu Formula high-dose group and IPM group had the most significant improvements (P<0. 05, P<0. 01). Conclusions Xuanfei Jiedu can significantly improve the general state, body weight, lung tissue W/ D, and lung tissue pathology, and the reduce inflammation and oxidative stress, of MDR-PA rats. The mechanism may be related to its inhibition of the expression of TLR4 / Myd88 / NF-κB pathway proteins in lung tissue.

Abstract: Objective This study was performed to establish and compare guinea pig models of tuberculosis using intranasal and aerosol infection routes at different doses. The overall goal was to provide a foundation for establishing a standardized guinea pig model of tuberculosis for the study of respiratory tract infection. Methods Twenty-four female guinea pigs were randomly divided into six groups of four guinea pigs each. They were then infected with two doses of Mycobacterium tuberculosis through either the aerosol route (groups A, B, and C) or intranasal route (groups D, E, and F). Aerosol infection groups consist of 3 groups : group A (Aerosol control group, uninfected control group), group B (Aerosol low-dose group, 5 × 10² CFU), and group C (Aerosol high-dose group, 5 × 10³ CFU) Intranasal infectiongroups also consist of 3 groups : group D ( Intranasal control group, uninfected control group), group E ( Intranasal lowdose group, 1 × 10⁴ CFU),and group F ( Intranasal high-dose group, 5 × 10⁴ CFU). The clinical manifestations of the guinea pigs were observed after infection. All guinea pigs were euthanized on day 14. Lung, spleen, and liver tissues were obtained for gross examination and histopathological analysis using hematoxylin-eosin staining to identify characteristic lesions associated with tuberculosis. Acid-fast staining was performed on in situ tissues and organs followed by bacterial culture to analyze the bacterial load. Results The guinea pigs in four infection groups ( B, C, E, and F) exhibited macroscopic tuberculosis lesions in the lung, spleen, and liver. Histopathological examination revealed the presence of tuberculous granuloma lesions. Acid-fast staining and bacterial load analysis demonstrated that the bacteria were primarily localized in the lung tissue of aerosol-infected groups B and C, with a few also present in the spleen and liver, and the bacterial load was 10⁴~ 10⁵ CFU/ mL. In intranasal infection groups E and F, bacteria were found in the lung, spleen, and liver with a similar bacterial load of 10⁴~ 10⁵ CFU/ mL. There was no significant difference in lesion severity or bacterial load among groups B, C, E, and F; however, groups B, C, and F showed low standard deviations for both pathology and etiology. Conclusions A guinea pig model of acute tuberculosis was successfully established using two doses administered through distinct routes of infection. Pathological examination and pathogenic analysis demonstrated that an aerosol dose of 5× 10² CFU of Mtb effectively established a homogeneous model of acute tuberculosis with good consistency among th animals. Additionally, intranasal infection with 5 × 10⁴ CFU of Mtb produced a relatively uniform model of tuberculosis.Notably, however, aerosol infection at 5 × 10² CFU progressed to an acute tuberculosis model more rapidly than intranasal infection at 5 × 10⁴ CFU

Abstract: Objective To optimize a C57BL/ 6J mouse liver fibrosis model induced by different doses of carbon tetrachloride through imaging, molecular biology, and pathology method. Methods Thirty-six healthy C57BL/ 6J male mice were randomly divided into a control group, 2 weeks, 3 weeks, 4 weeks, 6 weeks, and 8 weeks groups (n=6) after adaptive feeding for 1 week. The control group was intraperitoneally injected with 0. 2 mL olive oil three times a week, and the positive-control groups were intraperitoneally injected with 0. 2 mL 20% CCl₄-olive oil solution three times a week. Changes in the body weights of mice in each group were recorded. Liver stiffness was measured on days 15, 22, 29, 43 and 57, and blood samples were collected, and cereal third alanine aminotransferase ( ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), pro-type Ⅲ collagen (PC-Ⅲ), and type Ⅳ collagen (Ⅳ-C) content was measured. The liver tissues were stained with hematoxylin and eosin (HE), Masson, and Sirius red. The Metavir scoring system was used to evaluate the degree of liver fibrosis. Results Compared with the control group, mice in the positivecontrol groups were listless and tended to huddle together. In terms of body weight, the 4 weeks, 6 weeks, and 8 weeks groups were significantly lighter than the control group, while the 2 weeks group mice were significantly heavier than the control group mice. Liver elastography showed a progressive increase in stiffness with increased administration time. The biochemical tests showed that, compared with the control group, the other groups’ ALT and AST levels were significantly higher. With an increase in drug delivery time, the positive-control group’ s HA, LN, PC-Ⅲ and Ⅳ-C levels showed increasing trends. Pathological examination revealed that liver fibrosis was progressively aggravated with an increase in administration time. At 4 weeks, the pathological diagnosis was consistent with that of liver fibrosis, and there were signs of pseudolobule formation at 6 weeks. Pseudolobules were formed at 8 weeks, suggesting early cirrhosis. Conclusions A liver fibrosis model can be successfully established in C57BL/ 6J mice by intraperitoneal injection of 20% CCl₄ -olive oil solution three times a week for 4 consecutive weeks. The model has good stability, and the modeling method is rapid and can be used as an optimized scheme for the establishment of liver fibrosis models.

Abstract: Objective Using the concept of allostatic load ( AL) to evaluate aging of male SD rats and the effectiveness of Zuogui Pill in naturally aging rats. Methods Naturally aging male SD rats were tested at the ages of 2, 5,8, 14, 18, and 21 months. They were divided into an elderly control group, low-dose Zuogui Pill group, and high-dose Zuogui Pill group. Intervention with Zuogui Pill was trialed for 3 months. Blood samples were taken from the tails of rats each month, and the number of T lymphocytes and rate of apoptosis were measured by flow cytometry. Low-density lipoprotein cholesterol ( LDL-c), high-density lipoprotein cholesterol ( HDL-c), triglycerides ( TG), total cholesterol (TC), free fatty acids (FFA), 25 hydroxyvitamin D (25-OH-D), corticosterone (CORT), C-reactive protein (CRP),and interleukin-6 ( IL-6) were detected in rat sera. By identifying the collinearity between indicators and professional considerations, LDL-c, TC, HDL-c, FFA, TG, CORT, IL-6, CRP, 25-OH-D, CD3⁺T cell count, and CD3⁺T cell apoptosis rate were included in the AL scoring. The threshold for each indicator was established with data from 5-month-old rats, and the score was 1 point below or/ and above the threshold. Results The serum levels of LDL-c, TG, TC,25-OH-D, CRP, and IL-6 of rats showed significant changes with age, although the patterns of change differed. The CD3⁺T lymphocyte count significantly decreased with age (P<0. 01), while the apoptosis rates of CD3⁺, CD4⁺, and CD8⁺T lymphocytes significantly increased with age (P<0. 01). Zuogui Pill significantly increased serum CORT levels in elderly rats (P<0. 01) and reduced the apoptosis rate of CD8⁺T lymphocytes (P<0. 05). The AL score began to increase in rats at 5 months of age and reached its peak in those of 18 months of age. Conclusions AL can better characterize the aging process compared to a single indicator. Zuogui Pill can improve the stress response ability of aging rats and alleviate immunosenescence.

Abstract: Objective To investigate the role of LncRNA-gm33782 in tubular injury in acute kidney injury (AKI) and the mechanism by which isorhamnetin ameliorates inflammatory cell apoptosis of tubular cells in AKI. Methods Forty-eight male C57BL/ 6J mice were randomly assigned to four groups: control group, AKI model group, electroporation + AKI group, and treatment group ( oral administration of 30 mg / kg isorhamnetin). AKI was induced by a one-time intraperitoneal injection of 20 mg / kg cisplatin. Chromatin isolation by RNA purification was used to capture the LncRNAgm33782 binding protein in AKI-affected kidneys for mass spectrometry, revealing the direct target protein of LncRNAgm33782. The role of LncRNA-gm33782 in AKI was evaluated by observing the renal function, pathological structure, and expression of renal inflammatory factors ( interleukin-1β, interleukin-6, and tumor necrosis factor-α) in mice after electrotransfection and isorhamnetin treatment. Nuclear factor-κB was detected as a critical mediator of inflammation. The expression levels of Bax and Bcl-2 protein were detected and flow cytometry was performed to evaluate the therapeutic effect of isorhamnetin on tubular cell apoptosis in AKI. Results Kidneys with cisplatin-induced AKI showed severe renal tubule injury, macrophage infiltration, and inflammation. Isorhamnetin treatment and LncRNA-gm33782 electrotransfection knockdown alleviated these signs of AKI. LncRNA-gm33782 was mainly expressed in renal tubular cells with AKI. LncRNA-gm33782 binding protein was detected by mass spectrometry, and complement factor H was found to have a direct binding relationship with LncRNA-gm33782. After LncRNA-gm33782 was overexpressed in vitro, the expression of complement factor-H immediately increased, and the therapeutic effect of isorhamnetin on apoptosis of AKI-affected inflammatory cells was inhibited. Conclusions Isorhamnetin alleviates apoptosis of tubular inflammatory cells in AKI by inhibiting the regulatory effect of LncRNA-gm33782 on complement factor H.

Abstract:With increased population aging, the incidence of osteoporosis (OP) increases year by year, and has become a key global public health problem. Its main characteristics are reduced bone mass and damage to bone microstructure, often clinically manifesting as pain, hunched-back posture, height reduction, and even bone fractures.Therefore, it is particularly important to actively carry out scientific research related to OP, and the construction of animal models has become one of the key drivers of medical research, especially for exploring new treatments and in drug development. Animal models that are a “combination of disease and syndrome”, integrating the characteristics of Chinese medicine, occupy an irreplaceable position in research aiming to modernize traditional Chinese medicines. On the basis of the current research status of disease-syndrome combination OP animal models, this paper summarizes model construction and evaluation method to provide certain ideas and references for research using these specialized OP animal models.

Abstract:Audiogenic seizures (AGS) are the result of an epilepsy caused by strong acoustic stimulation and are characterized by generalized muscle spasms. AGS models are vital for studies of epileptogenesis, the search for causative genes and regulatory channels, and the screening of new antiepileptic drugs (AEDs). This paper summarizes the current progress of research on common animal models of AGS in terms of their pathogenetic features, possible pathogenesis, and causative genes to provide new research directions and targets for the development of AEDs.

Abstract:Duchenne muscular dystrophy (DMD) is a lethal, progressive, X-linked recessive hereditary muscle disease caused by a mutation in the gene encoding dystrophin. Currently, no cure for DMD is available, and clinical research is progressing slowly. The establishment of animal models is becoming increasingly important for experimental research on DMD. Following the research findings that mdx mice have the same pathogenesis as DMD patients, this model is widely used in the study of DMD pathogenesis and drug development. Mitochondrial injury is one of the most important pathological mechanisms of DMD, and mitochondrial protection is a potential therapeutic strategy for DMD, and thus it is significant to study mitochondrial injury in mdx mice. This article reviews the progress of research into mitochondrial injury in DMD model mdx mice in recent years to provide a reference for related experiments.

Abstract:Ferrets offer an advantage in nonclinical studies of anti-infective drugs because of their ability to be infected with and spread pathogenic microorganisms, especially viral strains, without the need for host adaptation.Additionally, the clinical symptoms exhibited by infected ferrets are very similar to those of humans. Although ferrets play a very important role in the research and development of antiviral drugs, the scope of their application remains limited. This may be related to the lack of corresponding national standards for laboratory animal feeding and application of ferrets as well as the lack of specific diagnostic and detection reagents. This paper summarizes the characteristics of ferrets as infectious disease models with a summary and analysis of the application direction of ferrets in anti-infective drug research. Our aim is to promote further standardization of the use of ferrets.