Vaccines are often administered by intramuscular or subcutaneous injection. Although these methods can effectively deliver vaccine antigens to the body and trigger the immune response. However, there are also some limitations, such as injection pain, complex operation, strict transportation conditions, poor immunogenicity and stability of the vaccine. With the development of technology, skin administration has become another new way to solve the above-mentioned problems of vaccine. Among the many ways of skin administration, microneedles show unique advantages and potential. This paper briefly describes the mechanism of microneedle transdermal immunity and its advantages over traditional injection. This paper briefly describes the mechanism and advantages of transdermal immunity of microneedles, enumerates the classification of microneedles, focuses on the design concept, structural advantages and preparation technology of bionic microneedles, and analyzes its application potential in vaccines. At the same time, this paper briefly describes the application of microneedle in vaccine, including bacterial infection, viral infection, cancer treatment and current clinical application progress. The challenges and prospects of microneedle vaccine are summarized from the aspects of safety, stability and acceptability of microneedle vaccine.

Conjugated estrogens (CE) are widely used in menopausal hormone therapy (MHT) for the relief of menopause-related symptoms (e.g., vasodilatory symptoms, neuropsychiatric symptoms, genitourinary atrophy symptoms, etc.) and the prevention of postmenopausal osteoporosis. This article reviews the characteristics, pharmacological effects, and practical applications of different dosage forms of CE (tablets, creams, and injections) in clinical practice. It has been shown that tablet CE effectively regulates the body's temperature center through systemic treatment, reduces hot flashes and night sweats, promotes bone formation, and increases bone density. Cream CE is used for local treatment to improve the genitourinary syndrome of menopause (GSM). Injectable CE is used for the treatment of abnormal uterine bleeding caused by hormonal imbalance without organ pathology. CE administered intravenously is indicated only for short-term use to rapidly and temporarily increase estrogen levels. Additionally, CE plays an important role in metabolism by improving insulin clearance and sensitivity and potentially protecting the nervous and cardiovascular systems, demonstrating a wide range of therapeutic potential. This paper provides new ideas for the design and development of drug dosage forms by exploring the formulation and application of CE.

Through literature analysis, the advantages of oral pharmaceutical vehicles in solving the temporary dispensing problems for special populations are summarized. Detailed introduction is made on the research, development, production, use, and regulatory experience of oral pharmaceutical vehicles in the United States, as well as the current research progress, achievements, and challenges faced by domestic oral pharmaceutical vehicles. The United States has formed a mature system in the field of oral pharmaceutical vehicles, and China can learn from the experience of the United States to improve relevant laws and policies, promote the establishment of technical guidance principles and technical standards for the use of oral pharmaceutical vehicles in temporary dispensing, and accelerate its clinical use.

Indigo naturalis [Baphicacanthus cusia (Nees) Bremek., QD], as a traditional Chinese medicine, has exhibited efficacy in the ulcerative colitis (UC). Cu Dian (CD), the current form of Indigo naturalis, has been regarded as the mainstream form of medicine today. Dian Hua (DH) is the traditional purified form of QD, in which the content of indigo and indirubin is higher than that of CD. The study evaluated the efficacy of DH and CD in UC and explored their mechanism. Male BALB/c mice were subjected to an 8-day regimen of 3% dextran sodium sulphate (DSS) drinking water to induce UC. The experiment was approved by the Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine (approval number: 2024075). Concurrently, the mice received intragastric administration of CD (400, 200, and 100 mg·kg^-1) and DH (400, 200, 100, and 50 mg·kg^-1) for the same duration. The anti-inflammatory properties of CD and DH were evaluated by quantifying levels of myeloperoxidase (MPO), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and IL-18 in colon tissue. Western blot and immunofluorescence assays were employed to assess the protein levels of zonula occludens 1 (ZO-1) and occludin. Additionally, Western blot and RT-qPCR were utilized to analyze the protein and gene expression levels of AMP-activated protein kinase (AMPK), nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3), and related factors in colon tissue. CD and DH demonstrated a capacity to alleviate inflammatory responses in mice with UC. The protective impact of both CD and DH on the intestinal mucosal barrier was associated with an elevation of ZO-1 and occludin. Furthermore, the anti-inflammatory effects of CD and DH were attributed to the inhibition of the NLRP3 inflammasome through the activation of the AMPK/silent information regulator of transcription 1 (SIRT1) pathway. Notably, DH exhibited a more pronounced improvement in UC compared to CD, particularly at the dosage of DH-M (200 mg·kg^-1). Our investigation substantiates the effectiveness of CD and DH in mitigating DSS-induced UC in mice. They demonstrated a capacity to diminish the production of inflammatory cytokines and safeguard the integrity of the intestinal epithelial barrier, notably by elevating level of tight junctions. The anti-colonic inflammatory effects of CD and DH were elucidated through the inhibition of both the formation and activation of the NLRP3 inflammasome, mediated by the AMPK/SIRT1 pathway.

To investigate the effect and underlying mechanism of action of Yueju volatile oil (YJVO) in the treatment of high altitude sleep disturbance (HASD) mice, gas chromatography-mass spectrometry (GC-MS) was used to identify the components of YJVO, while network pharmacology was applied to predict the mechanism of action. KM mice were selected and randomly assigned to several groups: the control group (control), the model group (model), and the YJVO treatment groups at low (YJVO-L, 200 μL·m^-3), medium (YJVO-M, 400 μL·m^-3), and high (YJVO-H, 800 μL·m^-3) doses, as well as the diazepam group (DZP, 2 mg·kg^-1). Except for the control group, all mice were subjected to a hypobaric oxygen chamber to establish the HASD model. Animal experiments were approved by the Animal Ethics Committee of Jiangxi University of Traditional Chinese Medicine (ethics No.: TEMPOR20230088). Co-sleeping and weight-bearing swimming experiments were conducted to assess the sleep-regulating effects of YJVO. Hematoxylin and eosin (H&E) staining was employed to observe damage in hypothalamic and hippocampal brain tissues. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure levels of melatonin (MT), γ-aminobutyric acid (GABA), interleukin-6 (IL-6), interleukin-1beta (IL-1β), 5-hydroxytryptamine (5-HT), orexin-A, and tumor necrosis factor-α (TNF-α) in both plasma and brain tissues. Concurrently, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were detected to evaluate oxidative stress. Western blot was used to determine the expression of key proteins in the NF-κB/NLRP3 pathway, and immunohistochemistry (IHC) was used to detect the expression of 5-hydroxytryptamine 1A receptor (5-HT_1A) in hippocampal and hypothalamic tissues. The GC-MS results revealed that YJVO was identified to contain 68 components. Network pharmacology results indicated that the mechanism of YJVO in treating HASD involves multiple signaling pathways such as AGE-RAGE, TNF, serotonin, and NF-κB. Behavioral experiments indicated that YJVO significantly prolonged the sleep duration, reduced the sleep latency, and bolstered the physical endurance and anti-fatigue capabilities of mice (P < 0.01). H&E staining results showed significant improvement in the pathological damage of the hippocampus and hypothalamus tissues in HASD mice. ELISA results indicated that YJVO increased the concentrations of sleep-inducing neurotransmitters MT and GABA within brain tissues, decreased the levels of wakefulness-inducing neurotransmitters orexin-A and 5-HT in plasma, and attenuated the secretion of pro-inflammatory cytokines IL-6, IL-1β, and TNF-α (P < 0.01). Biochemical results indicated that YJVO could inhibit the production of MDA in the brain tissue of HASD mice and enhance the activity of SOD and GSH-PX (P < 0.01). Western blot results showed that YJVO downregulated the protein expression of phosphorylate nuclear factor-kappa B (p-NF-κB p65)/nuclear factor-kappa B (NF-κB p65), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase 1 (Caspase-1), and IL-1β in the brain tissue of HASD mice (P < 0.01). IHC results demonstrated that YJVO downregulated the protein expression of 5-HT_1A in the hippocampus and hypothalamus tissues of HASD mice (P < 0.01). The study findings indicate that YJVO demonstrates potent therapeutic effects in HASD mice, potentially through the modulation of neurotransmitters and the attenuation of neuroinflammation.

This study investigates the anti-fatigue effects and mechanisms of Lycium barbarum and Ginseng combination using network pharmacology and in vivo validation. The effective components and their action targets of Lycium barbarum and Ginseng were explored through TCMSP, ETCM and other databases combined with literature. The fatigue targets were obtained through OMIM and Gene Cards databases. The intersection targets of drug targets and disease targets were screened out and imported into String database and Cytoscape 3.10.0 to construct PPI network. GO and KEGG enrichment analysis of the core targets were performed by David database. A mouse exercise-induced fatigue model was established to evaluate the anti-fatigue effects and mechanisms of the Lycium barbarum-Ginseng combination. The results showed that 55 active ingredients of Lycium barbarum and Ginseng were obtained, corresponding to 573 targets. The number of fatigue targets was 1 137, 115 total targets and 26 core targets were screened. KEGG enrichment pathways mainly included PI3K-AKT, HIF-1α, AGE-RAGE and other signaling pathways. All animal experiments were approved by the Experimental Animal Ethics Committee of Nanjing University of Chinese Medicine (approval number: 202308A018). The results showed that the low, middle and high dose groups of Lycium barbarum and Ginseng (1∶1) could prolong the exhaustive swimming time of mice, and the middle dose group had a more significant effect than Ginseng group and Lycium barbarum group. The middle and high dose groups of Lycium barbarum combined with Ginseng significantly reduced blood urea nitrogen (BUN) in mice. Compared with Ginseng group and Lycium barbarum group, the high dose group had a more significant effect. Lactic acid (LD) levels were significantly decreased in Ginseng group, Lycium barbarum group and combination group. Compared with the Ginseng group, the levels of liver glycogen (Lgly) and muscle glycogen (Mgly) were significantly increased in the middle dose group of Lycium barbarum combined with Ginseng. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were significantly decreased in Ginseng group and Lycium barbarum combined with Ginseng high dose group. The content of glutamic oxaloacetic transaminase (AST) and glutamic pyruvic transaminase (ALT) in the high dose group of Lycium barbarum combined with Ginseng can be significantly decreased. The results of Western blot showed that the low dose group of Lycium barbarum combined with Ginseng could significantly up-regulate the expression of P-PI3K and AKT proteins in the muscle tissue of fatigue mice, and the middle dose group could significantly down-regulate the expression of P-AKT and HIF-1α proteins. Compared with Ginseng group and Lycium barbarum group, the expression of PI3K protein in the high dose group was significantly increased. The expression of AKT protein in the low-dose group was significantly increased. In conclusion, Lycium barbarum combined with Ginseng has more positive effects on improving exercise endurance, reducing the accumulation of metabolites and improving glycogen storage levels in mice than Ginseng group and Lycium barbarum group, and its mechanism may be the regulation of AKT, PI3K, HIF-1α and other core targets and PI3K/AKT/HIF-1α signaling pathway to exert anti-fatigue effect.

Liver fibrosis is a common stage in the progression of chronic liver diseases, yet there is a lack of clinical drugs against liver fibrosis globally. Gynostemma pentaphyllum has the name of "Southern ginseng", commonly used in folk prevention and treatment of a variety of chronic liver disease, there are also reports of its anti-hepatic fibrosis. However, there are fewer relevant scientific studies. In this study, we used LC-MS metabolomics analysis to investigate the effects of Gynostemma pentaphyllum ethanol extract (GPE) on liver fibrosis in carbon tetrachloride (CCl₄)-induced mouse models and its potential mechanisms. All animal experiments were approved by the experimental animal ethics committee of Capital Medical University (DWLLGZR202202204). The results showed that GPE could significantly reduce the inflammatory cell infiltration and collagen accumulation in the liver of model mice, significantly reduce serum alanine transaminase and aspartate transaminase activity and hypoxanthine levels in mice, and could effectively inhibit the gene transcription and protein expression of collagen 1A1 (COL 1A1) and α-smooth muscle actin (α-SMA). Non-targeted metabolomics analysis of the liver showed that GPE mainly affected 47 differential metabolites; KEGG pathway enrichment analysis indicated that the differential metabolites were mainly enriched in the fructose/mannose metabolism and aromatic amino acid metabolism pathways. The targeted metabolomic assay was further used to validate a total of 13 differential metabolites of D-mannose, mannose 6-phosphate, D-fructose, fructose 2-phosphate, D-sucrose, trehalose, glutamate, phenylalanine, tyrosine, L-2-amino-3-oxobutyric acid, lactate, 2-hydroxybutyrate, and taurine, which may be important metabolites related to GPE's anti-fibrotic effects. This confirms that GPE's anti-liver fibrosis effects may be closely related to the regulation of the fructose/mannose metabolism pathway and the aromatic amino acid metabolism pathway.

To establish the cells stably co-expressing NOD1 receptor and enhanced green fluorescent protein (EGFP)-tagged nuclear factor of activated T cells 2 nuclear factor (NFAT2) (EGFP-NFAT2) in U2OS cell, the NOD1 (NM_006092) pcDNA3.1-3×Flag-hygro recombinant plasmid was transfected into U2OS-EGFP-NFAT2 cells, which were screened by pressure of hygromycin B and then incubated with NOD1 agonist lipopolysaccharides (LPS) for 30 min, and the green fluorescence intensity in the nucleus of the cells was detected by the high content screening assay. There were 46 cell strains expressing NOD1 in U2OS-EGFP-NFAT2 cells by EGFP-NFAT2 nuclear translocation assay. Among these cells, cells No 4 had the highest nuclear translocation function. Therefore, it was selected as the U2OS-EGFP-NFAT2-NOD1 cell for functional validation. The expression levels of NOD1 mRNA and protein in the selected U2OS-EGFP-NFAT2-NOD1 cells and the control cell U2OS-EGFP-NFAT2 were examined by real-time quantitative PCR (RT-qPCR) and Western blot. The results showed that NOD1 mRNA was stably expressed in this stably transfected cell line for 5-20 generations, and NOD1 protein was expressed in U2OS-EGFP-NFAT2-NOD1 stably transfected cell line, whereas no NOD1 protein was expressed in the control cell U2OS-EGFP-NFAT2. U2OS-EGFP-NFAT2-NOD1 cells were treated with histones or LPS for 30 min, and the EGFP-NFAT2 nuclear translocation was detected by the high content screening assay. Histones were found to significantly increase the EGFP-NFAT2 nuclear translocation in U2OS-EGFP-NFAT2-NOD1 stably transfected cells over a range of concentrations. The U2OS-EGFP-NFAT2-NOD1 cells were divided into the solvent control group, NOD1 receptor antagonist nodinitib-1+histone group, and histone group. The drug incubation time was 30 min, and the specificity of the NOD1 cells was verified by observing the EGFP-NFAT2 nuclear translocation through the high content screening assay. Compared with the histones group, the nodinitib-1+histones group significantly decreased EGFP-NFAT2 nuclear translocation in U2OSEGFP-NFAT2-NOD1 cells (P < 0.05). In conclusion, U2OS-EGFP-NFAT2-NOD1 cells stably co-expressing NOD1 and EGFP-NFAT2 are established, which can be used for screening antagonistic compounds targeting NOD1 pathogenic microorganisms with mechanism study.

Yeast-derived microcapsules were employed to co-encapsulate a nano-emulsion adjuvant (MF59) and antigens, effectively addressing the limitations of MF59 adjuvant in direct antigen encapsulation and its capacity to induce cellular immunity. Yeast microcapsules (YCs) were prepared using strong acid and alkali treatments, resulting in a porous and hollow structure with enhanced adjuvant properties. Positively charged polycaprolactone-polyethyleneimine (PCL-PEI) modified MF59 nanoemulsions were produced, which allowed for electrostatic interaction-driven spontaneous deposition into YCs. This modification facilitated the adsorption of the antigen, chicken ovalbumin (OVA), forming the complex YC-MF59-OVA. YC-MF59-OVA was efficiently recognized and endocytosed by antigen-presenting cells (APCs), a process facilitated by the β-glucan present on the capsular shell. Simultaneously, YC-MF59-OVA enabled a sustained release of the antigen and promoted the recruitment of APCs at the site of inoculation, leading to enhanced activation of immune responses in mice. Specifically, YC-MF59-OVA significantly elevated serum levels of IgG, IgG1, and IgG2a antibodies, achieving concentrations that were two to three times higher than those observed in the group treated with free OVA. In addition, the cellular immune response was notably improved, as evidenced by increased frequencies of IFN-γ⁺CD8⁺ and IL-4⁺CD4⁺ T cells compared to the OVA-immunized group. Furthermore, there was a marked increase in the proportion of memory T cells (CD44⁺CD62L⁺) in the splenic tissues of treated animals. The animal experiment protocol was reviewed and approved by Institutional Animal Care and Use Committee of Zunyi Medical University (approval No. ZMU21-2407-169). These findings demonstrate that YC-MF59-OVA can elicit robust humoral and cellular immune responses, confirming that YC can significantly overcome the limitations of traditional nanoemulsion adjuvants. This study provides a promising reference for the development of advanced vaccine delivery systems.

Severe alcoholic hepatitis (SAH) represents the most extreme form of alcoholic liver disease (ALD), accompanied by an extremely high mortality rate. Currently, there is a dearth of appropriate animal models for related research. The objective of this study is to establish a mouse model of SAH, thereby providing a preclinical animal model for subsequent research on SAH. This study is based on the NIAAA (National Institute on Alcohol Abuse and Alcoholism) model and constructs a mouse model by combining bacterial endotoxins. This experiment was approved by the Experimental Animal Ethics Committee of Capital Medical University (approval number: AEEI-2023-102). The model emulates the pathological processes of clinical SAH in terms of mouse mortality, liver tissue damage, and inflammatory markers, thereby establishing the model. Ultimately, it is ascertained that the optimal conditions for SAH mouse modeling based on the NIAAA model are the last intragastric administration of alcohol at a concentration of 7.5 g·kg^-1 in combination with intraperitoneal injection of lipopolysaccharide at a dose of 5 mg·kg^-1 for a period of 12 h. Under these conditions, the mouse model effectively simulates the high mortality and liver dysfunction seen in clinical SAH, with pathological staining results closely mirroring clinical findings. Additionally, it demonstrates a significant infiltration of neutrophils in the liver, indicative of an excessive inflammatory response. This model provides an ideal platform for preclinical research on SAH.