This article primarily investigates the role of the neural circuit from lateral hypothalamic (LH) glutamatergic neurons to the rostromedial tegmental nucleus (RMTg) in defensive behaviors in mice. Adult male vesicular glutamate transporter 2 (Vglut2)-Cre mice (8-10 weeks old) were randomly divided into a mCherry control group and a channelrhodopsin-2 (ChR2) experimental group, who received injections of AAV2/9-EF1α-DIO-mCherry and AAV2/9-EF1α-DIO-ChR2-mCherry virus into the LH, respectively. Optical fibers were implanted in the RMTg of both groups. After three weeks of recovery from surgery and sufficient viral expression, behavioral experiments were conducted, including the open field test, real-time place preference (RTPP) test, and hiding box test. After the experiment, ascertain the extent of viral infection and optical fiber positions in the mice. Immunohistochemistry was carried out to detect c-Fos expression in the RMTg to validate the activation of the pathway from LH glutamatergic neurons to the RMTg upon 473 nm blue laser stimulation. The results showed that c-Fos expression in the RMTg was elevated in ChR2-expressing mice following blue laser illumination, confirming successful pathway activation. During a 473 nm blue laser stimulation, compared to the mCherry group, mice in the ChR2 group exhibited a notable increase in velocity in the open field test, indicating an escape behavior. In the RTPP test, the time spent on the preferred side reduced, indicating avoidance behavior. Additionally, the latency to enter the hiding box significantly shortened, reflecting hiding behavior in the hiding box test. This research indicates that the activation of the projections from LH glutamatergic neurons to the RMTg promotes defensive behaviors in mice. All experimental procedures were approved by the Committee on the Ethics of Animal Experiments of the School of Basic Medical Sciences, Fudan University (identification number: 20250427-001).

Delayed fracture healing and nonunion continue to pose substantial challenges in orthopedic clinical practice. Existing medications for fracture treatment can, to a certain degree, promote fracture healing. However, they are plagued by problems such as unstable efficacy, slow onset, and frequent adverse effects. In this study, leveraging BanGel patented technology, we developed a novel Eucommia ulmoides-based hydrogel. This was achieved by integrating Eucommia ulmoides, a traditional Chinese medicine renowned for its liver and kidney tonifying and bone-strengthening properties, with a hydrogel delivery system. A transverse mid-shaft tibial fracture rat model was established. Behavioral assessments, serological analysis, radiological and histopathological examinations were employed to observe the dynamic changes during the fracture and subsequent healing processes. The efficacy of the Eucommia ulmoides hydrogel in promoting fracture healing was objectively evaluated. Moreover, the therapeutic advantages of the hydrogel were compared with those of oral and direct topical administrations containing the same dosage of Eucommia ulmoides. All animal experiments were approved by the Laboratory Animal Welfare Ethics Committee of the Laboratory Animal Center of the Institute of Basic Theory of Chinese Medicine, China Academy of Traditional Chinese Medicine, and were conducted in accordance with the relevant guidelines and regulations (approval No. IBTCMCACMS21-2406-03). The results showed that Eucommia hydrogel can significantly alleviate the early symptoms of fractured, promote callus formation, improve trabecular parameters and cortical bone continuity, and regulate the dynamic balance of bone formation and absorption. Compared with oral and topical applications of the same dose of Eucommia ulmoides, the Eucommia ulmoides hydrogel exhibited a faster onset and stronger permeability. Further mechanism exploration and verification results showed that Eucommia ulmoides can promote fracture healing by activating the serine/threonine protein kinase 1/β-catenin (AKT1/β-catenin) signaling axis to regulate the expression of Runt-related transcription factor 2 (RUNX2) and receptor activator of nuclear factor κB ligand/osteoprotegerin (RANKL/OPG). In summary, the novel Eucommia ulmoides hydrogel shows distinct advantages in promoting fracture healing and has potential clinical application value.

This study investigated the antitumor activity and molecular mechanisms of penfluridol (PF), an antipsychotic drug, in gastric cancer treatment. In vitro assays including CCK-8, colony formation, wound healing, Transwell migration, Hoechst 33342 staining, and flow cytometry demonstrated that penfluridol significantly inhibited proliferation and migration of SGC-7901 and HGC-27 gastric cancer cells in a concentration-dependent manner, while inducing apoptosis. Transcriptomic profiling combined with molecular docking revealed stable binding conformations between penfluridol and heat shock protein 90 (HSP90) or mouse double minute 2 (MDM2). Western blot analysis further confirmed that penfluridol suppressed the HSP90/protein kinase B/MDM2/tumor protein p53 (HSP90/Akt/MDM2/p53) pathway, characterized by decreased HSP90 expression and concomitant upregulation of p53 protein. These findings collectively indicate that penfluridol exerts multifaceted antitumor effects by targeting key oncogenic pathways, providing experimental evidence for repurposing this antipsychotic agent as a potential therapeutic strategy against gastric cancer.

Voltage-gated sodium channels (VGSCs, Na_V) are essential for the generation and conduction of action potentials in excitable cells. The Na_V1.8 subtype is predominantly expressed in sensory neurons, particularly in small-diameter neurons of the dorsal root ganglia (DRG), and plays a critical role in the signal transduction of chronic, neuropathic, and inflammatory pain. Due to its tetrodotoxin-resistant properties and electrophysiological characteristics of slow activation and inactivation, Na_V1.8 is a key target for analgesic drug development, facilitating repeated neuronal firing. Consequently, establishing an in vitro drug screening model based on Na_V1.8 is crucial for identifying lead compounds with analgesic potential. However, heterologous expression of Na_V1.8 remains challenging, primarily due to low expression levels in non-neuronal cell lines, which hinders effective drug screening. In this study, we established an in vitro expression model of rat Na_V1.8 using Xenopus laevis oocytes as the expression system. Channel property was characterized using the two electrode voltage clamp (TEVC) technique, providing a platform for screening active compounds. Quantitative polymerase chain reaction confirmed that rat Na_V1.8 is predominantly expressed in the DRG and trigeminal ganglia (Guangxi University Ethics Committee Review and approval number: GXU-2022-159). We cloned the full-length Na_V1.8 gene and constructed two chimeric variants, Na_V1.8-N and Na_V1.8-C. Following in vitro transcription of the corresponding capped RNA, it was injected into Xenopus laevis oocytes for heterologous expression. TEVC recordings were used to detect Na_V1.8 and chimera expression, and channel activity was evaluated through I-V curves, steady-state activation curves, and the specific Na_V1.8 antagonists A-803467 and VX-548. The chimera Na_V1.8-C resolved the problem in stable expression of Na_V1.8 in vitro and exhibited similar pharmacological properties with native Na_V1.8, offering a valuable tool for future screening of compounds targeting voltage-gated sodium channel.

Non-nucleoside reverse transcriptase inhibitors (NNRTIs) play an important role in AIDS treatment. Diarylpyrimidine derivatives (DAPYs) represent one of the most important classes of NNRTIs. The previously reported ZK-4a showed high cytotoxicity (CC₅₀ = 5.98 μmol·L^-1), significantly limiting its therapeutic potential. To resolve this issue, we adopted a two-pronged strategy: replacing the NH fragment with an O atom and modifying the C-5 position of the pyrimidine core. These modifications enabled the design and synthesis of 27 novel deuterated-biphenyl-DAPY analogues. Fortunately, all deuterated-biphenyl-DAPYs exhibited broad and highly efficient antiviral activity, as well as obviously decreased cytotoxicity (CC₅₀ = 18.02- > 297.27 μmol·L^-1). Among them, compound 8s achieved an EC₅₀ value of 8.88 nmol·L^-1 while showing negligible cytotoxicity (CC₅₀ > 287.68 μmol·L^-1). Compound 8s also displayed exceptional resistance profiles with EC₅₀ values of 7.54 nmol·L^-1 (L100I), 8.57 nmol·L^-1 (K103N), 24.28 nmol·L^-1 (Y181C), 62.71 nmol·L^-1 (Y188L) and 14.38 nmol·L^-1 (E138K). Additionally, the target of deuterated-biphenyl-DAPYs was verified by reverse transcriptase enzyme inhibition assays.

The 95% ethanol extract of llicium micranthum was separated and purified by silica gel, Sephadex LH-20, MCI-gel and semi-preparative HPLC. The chemical structures of the isolated compounds were identified by NMR and MS. Twelve compounds were isolated and identified as illimicranol A (1), vanilloloside (2), 4-O-β-D-glucosyl-benzoic acid (3), gastrodin (4), icariside D₂ (5), caryolane-1,9β-diol (6), clovane-2β,9α-diol (7), methyl shikimate (8), protocatechuic acid methyl ester (9), p-hydroxybenzyl alcohol (10), p-hydroxyphenylethanol (11), methyl p-hydroxyphenylacetate (12). Among them, compound 1 is a unique lignan with new carbon skeleton, named illimicranol A. Compounds 2-12 are firstly isolated from this plant. The effects of compound 1 on the differentiation of PC12 cells induced by NGF and the damage of PC12 cells induced by MPP⁺ were evaluated. The results showed that compound 1 did not exhibit obvious neurotrophic and neuroprotective activities.

Five compounds were isolated from the dichloromethane fraction of a 70% ethanol extract of Epimedium sagittatum Maxim using a combination of chromatography techniques, including silica gel column chromatography, ODS column chromatography, and semipreparative high-performance liquid chromatography techniques. The structures of these five compounds were identified as 5,4′-dihydroxy-3′-prenyl-7-prenyloxyflavanone (1), 5,4′-dihydroxy-7-prenyloxyflavanone (2), 5,7,4′-trihydroxy-3′,8-diprenylflavanone (3), sebacic acid (4), and aurantiamide acetate (5), respectively, based on a comprehensive analysis of their physicochemical properties and spectroscopic data, such as NMR and MS. Notably, compound 1 was found to be a new flavonone compound, while compounds 2-5 were isolated from the Epimedium genus for the first time. Additionally, preliminary investigations indicated that compounds 1 and 3 exhibited potential anti-pulmonary fibrosis activities.

Nineteen compounds were isolated from the ethyl acetate fraction of the stems and leaves of Salvia miltiorrhiza Bge by using various column chromatographic methods, such as silica gel, Sephadex LH-20 and semi-preparative HPLC. According to their physiochemical properties and spectral data, the structures were further identified as 3′,4′-dihydroxy-4-(3-methoxy-3-oxopropyl)-[1,1′-biphenyl]-3-carboxylic acid (1), 2-hydroxy-5-(2′-hydroxybenzyl) benzoic acid (2), dehydrozingerone (3), 3,4-dihydroxy benzalacetone (4), benzyl alcohol-β-D-glucopyranoside (5), dehydrololiolide (6), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]-oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (7), 6-hydroxy-3-(2-hydroxyethyl)-2,4,4-trimethylcy clohexa-2,5-dien-1-one (8), scopoletin (9), esculetin (10), dihydrotanshinone Ⅰ (11), 2′-deoxythymidine (12), quercetin 3-glucoside (13), sutchuenmedin A (14), 2′′-O-rhamnosylicariside-Ⅱ (15), 2,2′-oxybis(1,4-di-tert-butylbenzene) (16), 1H-indole-3-carbaldehyde (17), 3-(2-oxopropyl)-3-hydroxyind-olin-2-one (18), dysodensiol E (19). Compound 1 was a new biphenyl lignan, named as 3′,4′-dihydroxy-4-(3-methoxy-3-oxopropyl)-[1,1′-biphenyl]-3-carboxylic acid. Compound 2 was a new natural product and compounds 3-8 and 4-19 were firstly isolated from S. miltiorrhiza. MTT method was used to screen the anti-hepatoma activity of all the compounds, and none of them showed obvious effect.

The chemical profiling of Dengzhan Shengmai capsules were characterized using ultra high performance liquid chromatography/tandem quadrupole electrostatic field orbitrap (UHPLC-Q-Exactive-Orbitrap) high resolution mass spectrometry analysis. The post-acquisition strategy integrating precursor ion targeting recognition and diagnostic ion filtering was proposed to realize the identification of components in Dengzhan Shengmai capsules. The mass spectrometric data were acquired in both positive and negative ion modes using a full-scan MS1 coupled with data-dependent automatic triggering of MS/MS. By employing the similarity analysis function within the Access database software, precursor ions were promptly identified by comparing determined accurate masses to the in-house mass spectrometry database specific to Dengzhan Shengmai capsules. A diagnostic ion database encompassing a variety of compounds found in Dengzhan Shengmai capsules was compiled based on the mass spectrometry fragmentation patterns of 16 reference components. Ultimately, 248 compounds were characterized utilizing the diagnostic ion filtering method, comprising 51 caffeoyl esters, 38 flavonoids, 63 ginsenosides, 19 steroidal saponins, 11 nortriterpenoids, 21 homoisoflavones, 39 lignans and 6 other class constituents. The developed qualitative analysis approach enables swift and effective identification of chemical components present in Dengzhan Shengmai capsules, serving as a scientific basis for quality assessment and clinical utilization. This innovative strategy on high-resolution mass spectrometry data analysis, integrating precursor ion targeting recognition together with diagnostic ion filtering, introduced a novel methodology for the comprehensive analysis of complex substance systems in traditional Chinese medicine.

This study aimed to quantitatively evaluate the contents and in vivo exposure levels of 11 primary components in six commercially available Angong Niuhuang Pill (ANP) products. A chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous quantification of deoxycholic acid (DCA), cholic acid (CA), hyodeoxycholic acid (HDCA), glycocholic acid (GCA), taurocholic acid (TCA), curcumin (CC), baicalin (BC), berberine (BBR), and crocetin (CRT). A gas chromatography-mass spectrometry (GC-MS) method was established to determine muscone (MCN) and borneol (BN). The analytical methods were validated and applied to assess the content of the 11 components in six ANP products and their plasma exposure levels following intragastric administration in rats (All animal experiments had been approved by the Experimental Animal Ethics Committee of China Pharmaceutical University, ethics number: 2024-07-131). The quantitative methods utilizing LC-MS/MS and GC-MS were successfully established and validated for the simultaneous quantification of DCA, CA, HDCA, GCA, TCA, CC, BC, BBR, CRT, MCN and BN both in vivo and in vitro samples, respectively. The contents of HDCA, DCA, CA, GCA, TCA, CC, BC, CRT, BBR, BN and MCN varied over the ranges between 0.22 ± 0.03-1.47 ± 0.41, 0.22 ± 0.03-1.48 ± 0.39, 0.65 ± 0.01-4.28 ± 0.30, 0.44 ± 0.01-0.80 ± 0.02, 0.49 ± 0.05-1.36 ± 0.04, 0.001 2 ± 0.000 1-0.007 1 ± 0.001 0, 3.49 ± 0.80-4.47 ± 0.13, 0.003 6 ± 0.001 1-0.013 7 ± 0.014 8, 1.77 ± 0.05-4.84 ± 0.41, 4.70 ± 2.09-10.91 ± 3.08, 0.20 ± 0.03-0.44 ± 0.04 mg·g^-1, respectively. Similarly, the in vivo plasma exposure of HDCA, DCA, CA, GCA, TCA, BC, CRT covered the ranges over 5 088.63 ± 2 701.16-11 819.10 ± 7 585.61, 3 639.42 ± 1 970.63-18 853.46 ± 18 309.75, 19 397.89 ± 9 464.21-182 870.72 ± 236 816.06, 11 011.97 ± 4 236.99-57 331.79 ± 76 180.09, 53 607.57 ± 43 279.12-109 961.78 ± 82 288.37, 590.17 ± 175.72-2 424.90 ± 589.22, 74.95 ± 19.38-586.84 ± 194.49 h·ng·mL^-1, respectively. The plasma concentrations of the other four components (CC, BBR, BN, MCN) were below the detection limit. Three robust quantitative analytical methods were established and applied to evaluate variations of the content and in vivo exposure levels of 11 components in six ANP commercial products. The results demonstrated that the contents and in vivo exposure of some of the 11 components are consistent in the 6 products, particularly in the products containing the same natural or artificial bezoar and musk, or those manufactured by the same company. However, the inconsistency of the contents and in vivo exposure of some other ingredients suggests variations in the selection of raw herb medicines or processing technology. It suggests the crucial importance to monitor the source and quality of the raw herb medicines, and unify the processing technology. Furthermore, this study provides the data and reference methods for the supervision and consistency evaluation of quality amongst the products.