ObjectiveTo investigate the effect and mechanism of Sishenwan in restoring the intestinal barrier function in the rat model of diarrhea-predominant irritable bowel syndrome （IBS-D） due to spleen-kidney Yang deficiency based on intestinal flora and short-chain fatty acids. MethodsAfter the delivery of 10 SPF-grade pregnant rats， 4 male suckling rats were kept in each litter for the experiment. The male suckling rats were randomly allocated into blank， model， low-dose （3.51 g·kg^-1） Sishenwan， high-dose （7.02 g·kg^-1） Sishenwan， and Peifeikang （0.54 g·kg^-1） groups， with 8 rats in each group. The blank group was fed conventionally， and the other groups were subjected to mother-child separation and Sennae Folium gavage （1 g·mL^-1， 10 mL·kg^-1） for the modeling of IBS-D due to spleen-kidney Yang deficiency. After the modeling was completed， the rats in Sishenwan groups were administrated with the corresponding dose of Sishenwan decoction by gavage， and the Peifeikang group with bifidobacterium triple live powder+normal saline suspension. The blank and model groups were treated with an equal volume of normal saline by gavage. The general conditions and fecal characteristics of rats were observed. After 2 weeks of administration， the rats were anesthetized for sample collection. The pathological changes of the colon tissue in rats were observed by hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was employed to measure the levels of transforming growth factor-beta （TGF-β）， interleukin-10 （IL-10）， and interleukin-22 （IL-22）. Immumohistochemical staining （IHC） was performed to detect the positive expression of zonula occludens-1 （ZO-1） and occludin in the colon tissue. Western blot was employed to determine the protein levels of ZO-1 and occludin in the colon tissue of rats， and 16S rRNA gene sequencing was performed for intestinal flora. Gas chromatography-mass spectrometry was employed to determine the content of short-chain fatty acids （SCFAs） in the cecum contents of rats. ResultsThe colon tissue in the blank group presented a clear structure， neat glands， and no inflammatory cell infiltration. In the model group， the colon tissue showcased a disorganized structure， irregular arrangement of glands， and inflammatory cell infiltration. Compared with the model group， the low-dose and high-dose Sishenwan groups and the Peifeikang group exhibited an intact colon tissue structure， regular arrangement of glands， and reduced inflammatory cell infiltration. Compared with the blank group， the modeling lowered the levels of TGF-β， IL-10， and IL-22 in the serum （P<0.01）， down-regulated the protein levels of ZO-1 and occludin in the colon tissue （P<0.01）， and decreased the content of acetic acid and propionic acid and increased the content of butyric acid in cecum contents （P<0.05）. Compared with the model group， low-dose and high-dose Sishenwan raised the levels of TGF-β， IL-10， and IL-22 in the serum （P<0.05， P<0.01）， and Peifeikang elevated the levels of TGF-β and IL-10 in the serum （P<0.01）. High-dose Sishenwan and Peifeikang up-regulated the protein levels of ZO-1 and occludin （P<0.05， P<0.01）， increased the content of acetic acid and propionic acid in cecum contents （P<0.05）， and decreased the content of butyric acid （P<0.05）. The 16S rRNA gene sequencing results showed that the intestinal flora structure of the model group changed compared with that of the blank group. Compared with the model group， Sishenwan and Peifeikang increased the relative abundance of Lachnospiraceae， Muribaculaceae， Akkermansiaceae， Ligilactobacillus， UBA3282， Akkermansia， and Corynebacterium while reducing the relative abundance of Oscillospiraceae， Desulfovibrionaceae， Lactobacillus， Romboutsia， and Desulfovibrio. They can restore the intestinal flora structure similar to that in the blank group. ConclusionSishenwan can alleviate diarrhea symptoms and colonic mucosal inflammation， increase the expression of tight junction proteins in the colonic mucosa， and strengthen the intestinal barrier in IBS-D rats with the syndrome of spleen-kidney Yang deficiency. The mechanism of action may be related to optimizing the structure and balance of intestinal flora and regulating the SCFAs， and the effect of high-dose Sishenwan is obvious.

ObjectiveTo investigate the effect and mechanism of Sishenwan in restoring the intestinal barrier function in the rat model of diarrhea-predominant irritable bowel syndrome （IBS-D） due to spleen-kidney Yang deficiency based on intestinal flora and short-chain fatty acids. MethodsAfter the delivery of 10 SPF-grade pregnant rats， 4 male suckling rats were kept in each litter for the experiment. The male suckling rats were randomly allocated into blank， model， low-dose （3.51 g·kg^-1） Sishenwan， high-dose （7.02 g·kg^-1） Sishenwan， and Peifeikang （0.54 g·kg^-1） groups， with 8 rats in each group. The blank group was fed conventionally， and the other groups were subjected to mother-child separation and Sennae Folium gavage （1 g·mL^-1， 10 mL·kg^-1） for the modeling of IBS-D due to spleen-kidney Yang deficiency. After the modeling was completed， the rats in Sishenwan groups were administrated with the corresponding dose of Sishenwan decoction by gavage， and the Peifeikang group with bifidobacterium triple live powder+normal saline suspension. The blank and model groups were treated with an equal volume of normal saline by gavage. The general conditions and fecal characteristics of rats were observed. After 2 weeks of administration， the rats were anesthetized for sample collection. The pathological changes of the colon tissue in rats were observed by hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was employed to measure the levels of transforming growth factor-beta （TGF-β）， interleukin-10 （IL-10）， and interleukin-22 （IL-22）. Immumohistochemical staining （IHC） was performed to detect the positive expression of zonula occludens-1 （ZO-1） and occludin in the colon tissue. Western blot was employed to determine the protein levels of ZO-1 and occludin in the colon tissue of rats， and 16S rRNA gene sequencing was performed for intestinal flora. Gas chromatography-mass spectrometry was employed to determine the content of short-chain fatty acids （SCFAs） in the cecum contents of rats. ResultsThe colon tissue in the blank group presented a clear structure， neat glands， and no inflammatory cell infiltration. In the model group， the colon tissue showcased a disorganized structure， irregular arrangement of glands， and inflammatory cell infiltration. Compared with the model group， the low-dose and high-dose Sishenwan groups and the Peifeikang group exhibited an intact colon tissue structure， regular arrangement of glands， and reduced inflammatory cell infiltration. Compared with the blank group， the modeling lowered the levels of TGF-β， IL-10， and IL-22 in the serum （P<0.01）， down-regulated the protein levels of ZO-1 and occludin in the colon tissue （P<0.01）， and decreased the content of acetic acid and propionic acid and increased the content of butyric acid in cecum contents （P<0.05）. Compared with the model group， low-dose and high-dose Sishenwan raised the levels of TGF-β， IL-10， and IL-22 in the serum （P<0.05， P<0.01）， and Peifeikang elevated the levels of TGF-β and IL-10 in the serum （P<0.01）. High-dose Sishenwan and Peifeikang up-regulated the protein levels of ZO-1 and occludin （P<0.05， P<0.01）， increased the content of acetic acid and propionic acid in cecum contents （P<0.05）， and decreased the content of butyric acid （P<0.05）. The 16S rRNA gene sequencing results showed that the intestinal flora structure of the model group changed compared with that of the blank group. Compared with the model group， Sishenwan and Peifeikang increased the relative abundance of Lachnospiraceae， Muribaculaceae， Akkermansiaceae， Ligilactobacillus， UBA3282， Akkermansia， and Corynebacterium while reducing the relative abundance of Oscillospiraceae， Desulfovibrionaceae， Lactobacillus， Romboutsia， and Desulfovibrio. They can restore the intestinal flora structure similar to that in the blank group. ConclusionSishenwan can alleviate diarrhea symptoms and colonic mucosal inflammation， increase the expression of tight junction proteins in the colonic mucosa， and strengthen the intestinal barrier in IBS-D rats with the syndrome of spleen-kidney Yang deficiency. The mechanism of action may be related to optimizing the structure and balance of intestinal flora and regulating the SCFAs， and the effect of high-dose Sishenwan is obvious.

The development of antiviral drugs targeting SARS-CoV-2 remains a major strategy for the prevention and treatment of COVID-19 infection. Current anti-SARS-CoV-2 agents mainly act on key viral protein targets involved in the viral life cycle, such as the main protease (Mpro), papain-like protease (PLpro), and RNA-dependent RNA polymerase (RdRp). However, these drugs are insufficient to address the challenges posed by the rapid mutation rate and drug resistance of SARS-CoV-2. In recent years, viral RNA has demonstrated to be a highly promising therapeutic target. Small-molecule drugs that target viral RNA possess distinct advantages, featuring novel mechanisms of action and the potential for broad-spectrum antiviral activity as well as the ability to overcome drug resistance. Such agents provide a new avenue for combating coronavirus threats. This review systematically summarizes recent advances in the development of small-molecule inhibitors targeting conserved SARS-CoV-2 RNA structures, including the frameshift stimulation element (FSE), stem-loops (SL), G-quadruplexes (G-4), and RNA-targeting chimeras (RNATAC), aiming to provide some insight to guide future.

Objective:To investigate the role of heat shock transcription factor 1(HSF1)in hepatitis B virus X protein(HBx)-driven migration and invasion of hepatocellular carcinoma(HCC)cells,and to preliminarily explore the mechanism of HSF1 mediating HBx-driven HCC progression.Methods:4D label-free quantitative proteomics and Western blot were used to analyze the effect of HBx on HSF1 expression.HBx was overexpressed in the HCC cell lines Huh7 and MHCC97H,and its impact on the mRNA and protein levels and stability of HSF1 was assessed by RT-PCR and Western blot.The Cancer Genome Atlas database was used to analyze the expres-sion of HSF1 in hepatitis B virus(HBV)-associated HCC tissues and its relationship with tumor stage/grade and patient prognosis,and Western blot was used to measure the expression of HSF1 in HBV-associated HCC tissues.HBx was overexpressed in HCC cells,fol-lowed by HSF1 knockdown or cell treatment with the HSF1 inhibitor KRIBB11,and Transwell migration and invasion assay,scratch as-say,and F-actin staining experiment were performed to analyze the role of HSF1 in HBx-driven HCC cell migration and invasion.GEO and HCMDB datasets were used to identify the downstream target of HSF1,and the role of downstream target c-Myb in HSF1-me-diated HBx-driven HCC progression.Results:HBx upregulated HSF1 protein levels without significantly affecting its mRNA expres-sion,through enhancing HSF1 protein stability.HSF1 was highly ex-pressed in HBV-associated HCC tissues,and its elevated expres-sion correlated with tumor stage/grade and poor prognosis.HBx overexpression significantly promoted the migration,invasion,wound-healing capacity,and pseudopodia formation capacity of Huh7 and MHCC97H cells,while HSF1 knockdown or KRIBB11 treatment significantly attenuated the HBx-driven migration and in-vasion of HCC.HSF1 promoted the expression of the metastasis-associated protein c-Myb,and c-Myb overexpression in HSF1-knock-down HCC cells restored the promotive effect of HBx on HCC cell migration and invasion.Conclusion:HBx enhances HSF1 protein stability to promote its expression.Upregulation of c-Myb by HSF1 plays a pivotal role in HBx-driven HCC cell migration and inva-sion.Targeting HSF1 may help to delay the progression of HBV-associated HCC.

Oligodendrocytes(OLs)play a crucial role in the central nervous system,and the myelin sheath formed by them is essential for maintaining the normal function of the nervous system.Damage to the myelin sheath can trigger various nervous system diseases,which seriously affect the quality of life of patients.Clemastine,a traditional antihistamine,has shown certain potential in"repurposing old drugs"in recent years and can promote the repair of myelin sheath.This article summarizes the role of clemastine in regulating the differentiation,maturation,senescence,and apoptosis of OLs at multiple levels by acting on receptors on the OL membrane and regulat-ing the key signaling pathways in cells and epigenetic modifications in the nucleus.A deeper understanding of the mechanism of action of clemastine in OLs can help to provide new targets and new approaches for drug treatment of myelin-degenerative diseases.

Acute lung injury (ALI) has been a kind of acute and severe disease that is mainly characterized by systemic uncontrolled inflammatory response to the production of huge amounts of reactive oxygen species (ROS) in the lung tissue. Given the critical role of ROS in ALI, a Fe₃O₄ loaded bovine serum albumin (BSA) nanocluster (BF) was developed to act as a nanomedicine for the treatment of ALI. Combining with NIR irradiation, it exhibited excellent ROS scavenging capacity. Significantly, it also displayed the excellent antioxidant and anti-inflammatory functions for lipopolysaccharides (LPS) induced macrophages (RAW264.7), and Sprague Dawley rats via lowering intracellular ROS levels, reducing inflammatory factors expression levels, inducing macrophage M2 polarization, inhibiting NF-κB signaling pathway, increasing CD4⁺/CD8⁺ T cell ratios, as well as upregulating HSP70 and CD31 expression levels to reprogram redox homeostasis, reduce systemic inflammation, activate immunoregulation, and accelerate lung tissue repair, finally achieving the synergistic enhancement of ALI immunotherapy. It finally provides an effective therapeutic strategy of BF + NIR for the management of inflammation related diseases.

Working memory is an executive memory process that includes encoding, maintenance, and retrieval. These processes can be modulated by transcranial alternating current stimulation (tACS) with sinusoidal waves. However, little is known about the impact of the rate of current change on working memory. In this study, we aimed to investigate the effects of two types of tACS with different rates of current change on working memory performance and brain activity. We applied a randomized, single-blind design and divided 81 young participants who received triangular wave tACS, sinusoidal wave tACS, or sham stimulation into three groups. Participants performed n-back tasks, and electroencephalograms were recorded before, during, and after active or sham stimulation. Compared to the baseline, working memory performance (accuracy and response time) improved after stimulation under all stimulation conditions. According to drift-diffusion model analysis, triangular wave tACS significantly increased the efficiency of non-target information processing. In addition, compared with sham conditions, triangular wave tACS reduced alpha power oscillations in the occipital lobe throughout the encoding period, while sinusoidal wave tACS increased theta power in the central frontal region only during the later encoding period. The brain network connectivity results showed that triangular wave tACS improved the clustering coefficient, local efficiency, and node degree intensity in the early encoding stage, and these parameters were positively correlated with the non-target drift rate and decision starting point. Our findings on how tACS modulates working memory indicate that triangular wave tACS significantly enhances brain network connectivity during the early encoding stage, demonstrating an improvement in the efficiency of working memory processing. In contrast, sinusoidal wave tACS increased the theta power during the later encoding stage, suggesting its potential critical role in late-stage information processing. These findings provide valuable insights into the potential mechanisms by which tACS modulates working memory.
Humans ; Memory, Short-Term/physiology* ; Male ; Female ; Young Adult ; Transcranial Direct Current Stimulation/methods* ; Brain/physiology* ; Adult ; Electroencephalography ; Single-Blind Method

In November 2024， the Expert Group on Alcohol-related Liver Disease released a position statement on metabolic dysfunction and alcohol-related liver disease （MetALD）. MetALD is a new subtype of steatotic liver disease and refers to MASLD patients with a relatively large amount of alcohol consumption. The position statement points out the importance of accurate evaluation of alcohol consumption and recommends to quantify alcohol consumption using standard methods and alcohol biomarkers， and a comprehensive diagnosis should be made based on metabolic risk factors. In addition， the position statement analyzes the influence of drinking pattern on the diagnosis of MetALD and recommends to consider long-term drinking history during typing. The position statement also discusses the complex association between drinking and the diseases including metabolic syndrome， hepatic steatosis， fibrosis， and type 2 diabetes mellitus， and it is pointed out that the hierarchical management of patients should be optimized based on liver histological models and noninvasive models. The position statement elaborates on the definition of MetALD， drinking assessment， the interaction between alcohol use and metabolic dysfunction， and the methods for comprehensive management of MetALD， in order to facilitate learning and provide guidance for clinicians and researchers in clinical practice.

Background: Epimedii Folium is well known for its medicinal value. Four Epimedium species—Euphorbia brevicornu, E. sagittatum, E. pubescens, and E. koreanum—are the designated original plants of Epimedii Folium. Objective: The objective of this study is to facilitate the identification of the four Epimedium species and clarify their distributional responses to climate change. Methods: In this study, we assessed the genetic divergence of the four species and identified the molecular markers for species identification by using chloroplast genome sequences. Furthermore, we forecasted the distribution of potentially suitable regions of the four species Folium under climate change. Results: The authors obtained 26 chloroplast genome sequences of the four species and identified 1393 variable sites and 273 indel events. Genetic divergence analyses revealed that E. koreanum had long genetic distance from the other three species. Compared with the complete chloroplast genome, six hypervariable markers were discovered, and both rps4-trnL and ndhF were chosen as Epimedii Folium-specific DNA barcodes. Climate change is expected to influence the geographical distribution of the four Epimedium species, which were primarily found in China, South Korea, and Japan, leading to both expansion and contraction of their distribution ranges. Conclusion: Two identification markers were selected as the specific DNA barcodes for all four original plant species of Epimedii Folium. In addition, the shift of potential suitable area in various climate scenarios has been predicted. With the support of identification markers and the dynamics of suitable distribution areas, we are able to establish a foundation for the sustainable utilization of medicinal Epimedium resources in the future.

Background: Optimizing cultivation techniques for traditional Chinese medicine has become a crucial means to improve the quality of medicinal materials. Microbial agents, as environmentally friendly and efficient plant growth promoters and soil conditioners, have increasingly attracted attention in eco-agriculture research. Objective: Our understanding remains limited regarding how the application of microbial agents, alone or in combination, affects changes in the rhizosphere microbiome and its association with the bioactive components of medicinal materials. Methods: In this study, Epimedium pubescens Maxim. was employed as a model plant to examine the effects of 2 microbial agents(Paenibacillus mucilaginosus and Bacillus subtilis) applied individually and in combination on plant growth and the accumulation of bioactive components. Additionally, this study explored the relationship between the rhizosphere microbiome and plant development. Results: The application of microbial agents increased the yield of E. pubescens leaves by 20.30% to 33.66% and enhanced the total flavonol glycosides content by 11.40% to 29.94%. Meanwhile, microbial treatments reshaped the rhizosphere microbiome, promoted the enrichment of beneficial microorganisms (e.g., Frankia and Paenibacillus), suppressed phytopathogenic fungi such as Didymella and Scytalidium, and enhanced the stability of the soil microbial co-occurrence network. The partial least squares path model suggested that microbial agents not only directly impact the quality of medicinal herbs but also indirectly alter the accumulation of bioactive components by modulating the soil microbiome. Conclusion: These findings deepen our understanding of the relationship between medicinal plant quality and rhizosphere microbiomes as mediated by microbial agents. They also provide a basis for designing and manipulating synthetic microbial communities to promote sustainable development in eco-agriculture.