ObjectiveTo generate cynomolgus and rhesus monkey models carrying the Booroola fecundity (FecB) variant of bone morphogenetic protein receptor type 1B (BMPR1B) gene (designated BMPR1B^FecB) using adenine base edit technology and to explore genetic improvement strategies for enhancing the reproductive performance of laboratory macaques. MethodsFour single-guide RNAs (sgRNAs) were designed to target the BMPR1B c.746A>G locus in cynomolgus and rhesus monkeys. The sgRNAs were combined with the ABE8e-SpRY base editor, and the resulting editing complexes were microinjected into zygotes of both macaque species. Following in vitro culture of the injected embryos, nested PCR amplification was performed to detect the base-editing efficiency at the target site to screen for highly efficient sgRNAs. Subsequently, normally developing gene-edited embryos were transferred into the oviducts of surrogates. Seven days after birth, ear skin or peripheral blood samples were collected from the newborn monkeys for genotyping and mosaicism analysis, and the growth, development, and health status of the gene-edited monkeys were monitored. ResultsAmong the four candidate sgRNAs, sgRNA4 was identified as achieving 100% editing efficiency at the c.746A>G site. A total of 17 live monkey offspring were obtained via embryo transfer, including 8 cynomolgus monkeys and 9 rhesus monkeys. Genotyping results showed that all 8 newborn cynomolgus monkeys carried the c.746A>G point mutation, with an editing efficiency of 100% (8/8), of which 37.50% (3/8) were mosaics. Among the 9 newborn rhesus monkeys, 8 individuals carried the c.746A>G mutation, with an editing efficiency of 88.89% (8/9), of which 75.00% (6/8) were mosaics. ConclusionThe ABE8e-SpRY base-editing system enables precise editing at the c.746A>G (p.Q249R) locus of the BMPR1B gene in both cynomolgus and rhesus monkey embryos, resulting in the successful establishment of laboratory monkey models carrying the BMPR1B^FecB variant. These models provide a valuable foundation for future studies of the regulatory role of the BMPR1B^FecB variant in reproductive performance at the non‑human primate level, as well as for exploring genetic improvement strategies to enhance reproductive performance in laboratory monkeys.

Uveal melanoma(UM)is a rare but highly aggressive intraocular malignancy. Although it accounts for less than 5% of all melanoma cases, UM is the most common primary intraocular malignancy in adults. In recent years, advances in molecular targeted therapies and immunomodulatory strategies have provided new opportunities for the clinical management of UM. This review summarizes the pathogenesis, molecular biomarkers, and major therapeutic advances in UM, with particular emphasis on analyzing the research achievements and clinical application prospects of signaling pathway-targeted inhibitors, immune checkpoint inhibitors, and combination treatment strategies. The potential value of individualized treatment and the limitations of current therapeutic approaches are also discussed. Current evidence suggest that multidisciplinary intervention and precision molecular diagnosis and treatment may significantly improve the prognosis of patients with UM, providing references for future clinical research and the optimization of therapeutic strategies.

Background: Inhalation-based combination therapy has gained increasing attention for local treatments. However, a key challenge remains in ensuring the sustained pulmonary release of multiple active ingredients, particularly in traditional Chinese medicine (TCM) formulations. Objective: This study investigates a novel PulmoSphere-based inhalable carrier designed for the sustained pulmonary release of multiple active ingredients, using Shuang-Huang-Lian as a model TCM formulation containing three chemical markers. Materials and methods: The carrier was developed using PulmoSphere technology, incorporating phospholipid complexes of the chemical markers and hyaluronic acid (HA) into spray-dried microparticles. The aerodynamic properties, release characteristics, pulmonary distribution, and anti-inflammatory efficacy of different formulations were evaluated in vitro and in mice. Results: The microparticles retained the excellent aerodynamic properties of conventional PulmoSphere particles, with a mass median aerodynamic diameter of approximately 3.1 μm and a fine particle fraction of approximately 55%. Compared to free Shuang-Huang-Lian or phospholipid complex-loaded PulmoSphere particles, the HA-containing particles prolonged the retention of chemical markers in the lung epithelial lining fluid, demonstrating sustained release in vivo. Additionally, the HA-containing formulation enhanced the exposure of the three chemical markers to immune cells and lung tissues, leading to improved and prolonged anti-inflammatory effects, even at decreased doses. Conclusion: This novel inhalable particle system represents a promising approach for sustained pulmonary co-delivery of multiple active ingredients, offering enhanced and extended local therapeutic efficacy.

ObjectiveTo explore the potential mechanism of action of the combination of Sanguisorbae Radix-Sophorae Flos （DH） in the treatment of ulcerative colitis （UC） using network pharmacology methods and molecular docking technology. MethodsNetwork pharmacology analysis was utilized to predict the potential targets of DH for the treatment of UC. The therapeutic effects were experimentally validated by inducing a UC model in mice with 3% dextran sulfate sodium （DSS）. The experimental groups were the normal group， the model group， the salazosulfapyridine group （100 mg·kg^-1）， and the low， medium， and high dose groups of DH （1.2， 2.4， and 4.8 g·kg^-1）. The efficacy of the treatment was assessed through the general condition of the mice， histopathological examination， and the expression levels of inflammatory markers in the colon. The effect of DH on angiogenesis was explored by messenger RNA （mRNA） detection of colonic angiogenesis-related mediators， vascular endothelial growth factor （VEGF） immunohistochemistry， microvessel density （MVD） detection， and transmission electron microscopy. The phosphatidylinositol-3-kinase （PI3K）-protein kinase B （Akt） signaling pathway proteins were quantitatively analyzed through Western blot to assess whether the suppression of pathological angiogenesis by DH is associated with this pathway. ResultsNetwork pharmacological analysis yielded 112 potential core therapeutic targets for the treatment of UC with DH， of which the core targets were tumor protein 53 （TP53）， JUN， interleukin （IL）-6， Akt1， and tumor necrosis factor （TNF）. Compared with the normal group， mice in the model group showed significant weight loss， colon shortening， and high DAI score， increased expression of inflammatory factors IL-6， IL-1β， and TNF-α， as well as increased mRNA expression levels of angiogenesis-related mediators VEGF， vascular cell adhesion molecule 1 （VCAM1）， angiotensin 1 （Ang1）， matrix metalloproteinase （MMP）-1， MMP-2， and MMP-9. The positive expression of CD31 and VEGF in colonic tissue increased， and the protein expression of the PI3K/Akt pathway was increased （P<0.05）. The endothelial cells of the colonic mucosa and the colonic vasculature were severely damaged. Compared with the model group， mice in the DH groups had significantly reduced weight loss and colon shortening， lower DAI scores， and a significant decrease in mRNA expression of inflammatory factors and angiogenesis-related mediators. In addition， there was decreased positive expression of CD31 and VEGF in colonic tissue and decreased protein expression of the PI3K/Akt pathway （P<0.05）. ConclusionNetwork pharmacology， molecular docking， and experimental validation are applied to explore the mechanism of action of DH in the treatment of UC， and it is found that DH is able to improve the symptoms of colitis and inhibit the pathological angiogenesis in UC mice. Its action might be related to affecting the PI3K/Akt pathway.

ObjectiveTo explore the potential mechanism of action of the combination of Sanguisorbae Radix-Sophorae Flos （DH） in the treatment of ulcerative colitis （UC） using network pharmacology methods and molecular docking technology. MethodsNetwork pharmacology analysis was utilized to predict the potential targets of DH for the treatment of UC. The therapeutic effects were experimentally validated by inducing a UC model in mice with 3% dextran sulfate sodium （DSS）. The experimental groups were the normal group， the model group， the salazosulfapyridine group （100 mg·kg^-1）， and the low， medium， and high dose groups of DH （1.2， 2.4， and 4.8 g·kg^-1）. The efficacy of the treatment was assessed through the general condition of the mice， histopathological examination， and the expression levels of inflammatory markers in the colon. The effect of DH on angiogenesis was explored by messenger RNA （mRNA） detection of colonic angiogenesis-related mediators， vascular endothelial growth factor （VEGF） immunohistochemistry， microvessel density （MVD） detection， and transmission electron microscopy. The phosphatidylinositol-3-kinase （PI3K）-protein kinase B （Akt） signaling pathway proteins were quantitatively analyzed through Western blot to assess whether the suppression of pathological angiogenesis by DH is associated with this pathway. ResultsNetwork pharmacological analysis yielded 112 potential core therapeutic targets for the treatment of UC with DH， of which the core targets were tumor protein 53 （TP53）， JUN， interleukin （IL）-6， Akt1， and tumor necrosis factor （TNF）. Compared with the normal group， mice in the model group showed significant weight loss， colon shortening， and high DAI score， increased expression of inflammatory factors IL-6， IL-1β， and TNF-α， as well as increased mRNA expression levels of angiogenesis-related mediators VEGF， vascular cell adhesion molecule 1 （VCAM1）， angiotensin 1 （Ang1）， matrix metalloproteinase （MMP）-1， MMP-2， and MMP-9. The positive expression of CD31 and VEGF in colonic tissue increased， and the protein expression of the PI3K/Akt pathway was increased （P<0.05）. The endothelial cells of the colonic mucosa and the colonic vasculature were severely damaged. Compared with the model group， mice in the DH groups had significantly reduced weight loss and colon shortening， lower DAI scores， and a significant decrease in mRNA expression of inflammatory factors and angiogenesis-related mediators. In addition， there was decreased positive expression of CD31 and VEGF in colonic tissue and decreased protein expression of the PI3K/Akt pathway （P<0.05）. ConclusionNetwork pharmacology， molecular docking， and experimental validation are applied to explore the mechanism of action of DH in the treatment of UC， and it is found that DH is able to improve the symptoms of colitis and inhibit the pathological angiogenesis in UC mice. Its action might be related to affecting the PI3K/Akt pathway.

Hyperuricemia is a chronic metabolic disease resulting from purine metabolic dysfunction and is classified under the category of"blood turbidity"in traditional Chinese medicine.Our team termed it"acid turbidity,"and its pathogenesis is closely related to the dynamic evolution of the clear and the turbid components.With the change of modern people's diet structure,the incidence of hyperuricemia is increasing annually owing to the intake of fatty,sweet foods and alcohol.Therefore,this paper explores hyperuricemia from the"indigestion of spleen and stomach"theory.The core pathogenesis of hyperuricemia is indigestion of spleen and stomach,the inversion of clear and turbid substances,and endogenous acid turbidity.The initial manifestation of hyperuricemia is the internal retention of acid turbidity and ascending-descending disharmony;the gradual manifestation of this disease is that indigestion causes heat,and acid turbidity transforms into poison;the final manifestation of this disease is that secular indigestion causes deficiency and the inversion of clear and turbid substances.It can be summarized into three syndromes:syndromes of internal retention of dampness-turbidity,dampness-heat toxin amassment,and dampness-heat due to spleen deficiency.Therefore,this paper proposes to treat the disease according to different syndromes,with ascending the clear and descending the turbid as the core of treatment.And the therapeutic approach employs the flexible application of three methods:transportation,resolving,and transformation.For syndrome of internal retention of dampness-turbidity,treatment focuses on promoting spleen transportation to eliminate dampness;for syndrome of dampness-heat toxin amassment,the strategy is to resolve indigestion and purge heat;and for syndrome of dampness-heat due to spleen deficiency,the aim is to resolve turbidity and clear heat.By ascending the clear and descending the turbid,so that"returning the clear and the turbid to the original,"the spleen and stomach regain harmony,functions of ascending and descending are reestablished,and hyperuricemia can be effectively managed.

OBJECTIVE To explore the effects of Lycium barbarum polysaccharides on the proliferation， migration， and immune escape of oral cancer cells by regulating the T cell immunoglobulin and mucin domain-containing protein 3 （TIM3）/ galectin-9 （Gal-9） signaling pathway. METHODS Human oral cancer cells KB and CAL27 were assigned to control group， L. barbarum polysaccharides low-concentration group （200 μg/mL）， L. barbarum polysaccharides high-concentration group （400 μg/mL）， pcDNA-NC group （transfection of pcDNA-NC plasmid）， pcDNA-TIM3 group （transfection of pcDNA-TIM3 plasmid）， high concentration of L. barbarum polysaccharides+pcDNA-NC group （400 μg/mL L. barbarum polysaccharides + transfection of pcDNA-NC plasmid）， and high concentration of L. barbarum polysaccharides+pcDNA-TIM3 group （400 μg/mL L. barbarum polysaccharides + transfection of pcDNA-TIM3 plasmid）. The proliferation， migration and invasion abilities of the cells， T cell killing rate as well as the levels of interferon-γ （IFN-γ） and interleukin-2 （IL-2） in the cell supernatant were measured. mRNA expressions of TIM3 and Gal-9 and protein expressions of indoleamine 2，3-dioxygenase 1 （IDO1）， programmed death-ligand 1 （PD-L1）， TIM3 and Gal-9 in the cells were also determined. RESULTS Compared with control group， the clone formation rate， scratch healing rate， the number of invasive cells， protein expressions of IDO1 and PD-L1， mRNA and protein expressions of TIM3 and Gal-9 in both cell types of L. barbarum polysaccharide low- and high-concentration groups were decreased significantly （P＜0.05）， while the proliferation inhibition rate， T cell killing rate， and the levels of IFN-γ and IL-2 were significantly increased （P＜0.05）. Compared with control group and pcDNA-NC group， the clone formation rate， scratch healing rate， the number of invasive cells， and protein expressions of IDO1 and PD-L1， mRNA and protein expressions of TIM3 and Gal-9 in both cell types of the pcDNA-TIM3 group were all significantly increased （P＜0.05）， while the proliferation inhibition rate， T cell killing rate， IFN-γ and IL-2 levels were significantly decreased （P＜0.05）. Compared with L. barbarum polysaccharides high-concentration group and high concentration of L. barbarum polysaccharides+pcDNA-NC group， the clone formation rate， scratch healing rate， the number of invasive cells， and protein expressions of IDO1 and PD-L1， mRNA and protein expressions of TIM3 and Gal-9 in both cell types of high concentration of L. barbarum polysaccharides+pcDNA-TIM3 group were significantly increased （P＜0.05）， while the proliferation inhibition rate， T cell killing rate， and the levels of IFN-γ and IL-2 were significantly decreased （P＜0.05）. CONCLUSIONS L. barbarum polysaccharides may inhibit the proliferation， migration， and immune escape of oral cancer cells by suppressing TIM3/Gal-9 signaling pathway.

The integrated medical and elderly care model provides comprehensive medical and elderly care services by establishing medical facilities in elderly care institutions or forming close cooperative relationships with surrounding medical institutions. Currently， there are 87，000 paired partnerships established nationwide between medical and health institutions and elderly care service institutions， and more than 7，800 integrated medical and elderly care institutions have obtained the qualifications of medical and health institutions and completed elderly care service registration. This model not only meets the elderly’s healthcare needs but also provides life care， psychological support， and social activities， thereby improving their quality of life. However， the integrated medical and elderly care model also faces numerous ethical dilemmas in practice. This paper aimed to explore in depth the ethical dilemmas and countermeasures optimization in the work of elderly care institutions under this model， to promote the improvement of service quality， comprehensively guarantee the rights and interests of the elderly， and promote the healthy development of this model in practice. Under this model， the elderly care institutions not only bear the responsibility of providing long-term care and life services but also need to cooperate with medical institutions to provide healthcare and health management services for the elderly. By exploring the practical paths for elderly care institutions to address the ethical dilemmas faced with this model， feasible solutions were proposed to enhance the welfare of the elderly and promote the harmonious development of society.

OBJECTIVE: To observe the effects of moxibustion at "Shenque" (CV8) on urodynamics and the expression of brain-derived neurotrophic factor (BDNF) and immediate early gene (c-fos) in pontine micturition center (PMC), periaqueductal gray (PAG), medial prefrontal cortex (mPFC) of neurogenic bladder (NB) rats after spinal cord injury. METHODS: Twenty-four SPF female SD rats were randomly divided into a sham-operation group (6 rats) and a modeling group (18 rats). In the modeling group, T₉ complete spinal cord transection method was used to establish a neurogenic detrusor overactivity model, and the 12 rats with successful modeling were randomized into a model group and a moxibustion group, with 6 rats in each group. The rats in the moxibustion group were treated with ginger/salt-insulated moxibustion at "Shenque" (CV8), and 4 consecutive moxa cones were delivered in one intervention. Moxibustion was operated once daily and for 14 days. After intervention completion, the urodynamic indexes of rats in each group were detected. Fluorescence quantitative PCR was used to detect the mRNA expression of BDNF and c-fos in PMC, PAG and mPFC in rats. Western blot was used to detect the protein expression of BDNF and c-fos in PMC, PAG and mPFC. RESULTS: The rats in the sham-operation group did not show phasic detrusor contraction during bladder filling. Compared with the model group, the frequency and amplitude of the phasic detrusor contraction were reduced 5 min before urine leakage in the rats of the moxibustion group (P<0.05), and the duration of the first phasic detrusor contraction during bladder filling was prolonged (P<0.05). Compared with the sham-operation group, the mRNA and protein expression of BDNF and c-fos in PMC, PAG and mPFC increased in the model group (P<0.05). Compared with the model group, the mRNA and protein expression of BDNF and c-fos in PMC, PAG and mPFC decreased in the moxibustion group (P<0.05). CONCLUSION Moxibustion at "Shenque" (CV8) can improve the phasic contraction during bladder filling in NB rats after spinal cord injury, possibly by down-regulating the mRNA and protein expression of BDNF and c-fos in PMC, PAG, and mPFC.
Animals ; Moxibustion ; Female ; Rats ; Brain-Derived Neurotrophic Factor/metabolism* ; Rats, Sprague-Dawley ; Acupuncture Points ; Spinal Cord Injuries/metabolism* ; Urinary Bladder, Neurogenic/etiology* ; Proto-Oncogene Proteins c-fos/metabolism* ; Humans ; Urinary Bladder/physiopathology* ; Brain/metabolism* ; Urination