Short-lasting unilateral neuralgiform headache attacks （SUNHA） are a rare type of disabling primary headache within the category of trigeminal autonomic cephalalgias （TACs）， and it has two subtypes of SUNCT （with conjunctival injection and tearing） and SUNA （with other autonomic features）. SUNHA is characterized by severe unilateral （often V1） stabbing/shock-like pain （lasting for 1-600 s）， high frequency （2‒600 attacks a day）， and prominent ipsilateral cranial autonomic symptoms （such as conjunctival injection，tearing， and nasal obstruction）. Trigger factors are observed in 86% of patients. The diagnosis of SUNHA should meet the ICHD-3 criteria （≥20 attacks）， and brain MRI （especially for the pituitary gland/posterior cranial fossa） should be performed to exclude secondary causes （such as neurovascular conflict and pituitary tumor）. Lamotrigine is used as first-line prophylaxis， while lidocaine aids acute relief in the transitional phase； occipital nerve stimulation， deep brain stimulation， or microvascular decompression can be used for refractory cases. It is of great importance to enhance awareness， achieve precise differentiation（from trigeminal neuralgia or other types of TACs）， and provide individualized treatment.

Image 1.

Objective:To analyze and identify the expression profiles of oxidative stress-related genes and circular RNAs(circRNAs)in ricin toxin(RT)-induced pyroptosis of mouse mononuclear macrophages(RAW264.7)using transcriptome sequencing and bioinformatics technology,and to preliminarily analyze their potential functions.Methods:The macrophages(RAW264.7 cells)were treated with RT to establish a cell pyroptosis model and divided into control group,40 μg·L-1 RT group,and 80 ng/mL RT group.Transmission electron microscope(TEM)was used to observe the morphology of the RAW264.7 cells in various groups;Western blotting method was used to detect the expression levels of the pyroptosis pathway-related proteins in the cells in various groups;80 μg·L-1 RT was selected for subsequent experiments.Transcriptome sequencing(RNA-Seq)was performed to obtain the circRNA and mRNA expression profiles in the RAW264.7 cells in control group and RT-treated group,followed by bioinformatics analysis.Results:Compared with control group,the cells in 40 and 80 μg·L-1 RT groups underwent morphological changes;the cells in RT groups showed obvious pyroptosis-like morphological changes,characterized by significant swelling of dying cells and the appearance of characteristic large bubbles on the plasma membrane.Compared with control group,the expression level of gasdermin DN-terminal fragment(GSDMD-N)protein in 40 and 80 μg·L-1 RT groups was increased(P<0.05);compared with 40 μg·L-1 RT group,the expression level of GSDMD-N protein in the cells in 80 μg·L-1 RT group was increased(P<0.05);therefore,the subsequent experiments used the RT concentration of 80 μg·L-1.A total of 2930 differentially expressed messenger RNAs(mRNAs)and 24 differentially expressed circRNAs were identified.The constructed circRNA-microRNA(miRNA)-mRNA competing endogenous RNA(ceRNA)regulatory network consisted of 7 circRNAs,12 miRNAs and 13 mRNAs.Gene Ontology(GO)functional enrichment analysis showed that in biological process(BP),the functions regulated by differentially expressed genes mainly included immune response and oxidative stress response;in cellular component(CC),differentially expressed genes were mainly localized to the external side of plasma membrane and cell leading edge;in molecular function(MF),they were mainly involved in transporter transmembrane activity and hormone receptor binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that differentially expressed genes were mainly enriched in Toll-like receptor signaling pathway,Forkhead box O(FoxO)signaling pathway and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)signaling pathway.In the protein-protein interaction(PPI)network,the top 10 hub genes with the highest connectivity were screened by CytoHubba,including matrix metalloproteinase 9(MMP9),superoxide dismutase 2(SOD2),and v-src sarcoma viral oncogene homolog(Src).Conclusion:The expression profiles of oxidative stress-related genes and circRNAs in RAW264.7 cells are altered after RT treatment.The screened differentially expressed circRNAs and mRNAs may serve as potential targets to regulate RT-induced pyroptosis in RAW264.7 cells through oxidative stress pathways.

Objective:To systematically analyze the registration status of clinical trials related to inflammatory bowel disease (IBD) in China Clinical Trial Registry (ChiCTR); To focus on the characteristics and shortcomings of TCM research; To provide data support and theoretical basis for optimizing clinical trial design and improving the quality of TCM research.Methods:The IBD-related clinical trials registered by ChiCTR from the establishment of the database to September 18, 2024 were retrieved. SPSS 26.0 software was used to analyze the frequency of research objects, registration time, registration area and institution, source of funds, research type and design scheme, random method and blind method, trial staging and research center, sample size, intervention measures and outcome indicators.Results:There were 317 clinical trials of IBD. Shanghai, Jiangsu, Beijing, Guangdong and Zhejiang accounted for 72.87% of the total number of registrations. Most of the registered projects were intervention studies (51.42%), 48 studies used blind method, and randomized controlled study was the main research design type. In the 68 clinical trials related to TCM, the intervention measures were divided into 4 categories, of which Chinese materia medica was the most (42 items); the sample size was the most in the intervention study, with a total of 6 787 cases; the total frequency of outcome indicators was 1 866 times, and the quality of life and mental health were the most (147 items).Conclusions:The number of registered IBD clinical trials is generally increasing, but there may be problems such as uneven distribution of regions and institutions, poor design of sample size, blind method and other research, and non-standard filling of registration information. In the research of TCM treatment of IBD, it is suggested to further strengthen the depth and breadth, especially the characteristic therapy of TCM.

Objective To enhance the cyclic peptide compound's membrane permeability,structural stability,and neuroprotective activity,based on the amino acid sequence of peptides of Tat-GluA2-3Y,by designing and synthesizing a serial of cyclic peptides through strategies including polypeptide cyclization,replacement of the cell-penetrating peptide,substitution with D-amino acids,and incorporation of mini polyethylene glycol fragments.Methods The target peptides were synthesized based on standard Fmoc solid-phase method,followed by analysis and purification via reverse phase high-performance liguid chromatography(RP-HPLC).The cytoprotective activity of the peptides was evaluated by using the HT-22 cell model.The transmembrane transport efficiency of the peptides was determined based on the Caco-2 monolayer intestinal epithelial cell model.Plasmatic plasma and metabolic stability of the peptides were measured by in vitro co-incubation experiments with rat plasma and human liver microsomes.Finally,the in vivo neuroprotective activity of the peptides was validated by using a mouse middle cerebral artery occlusion model.Results Seven cyclic peptides were successfully designed and synthesized by using the standard Fmoc solid-phase method,with purities exceeding 90%as confirmed by RP-HPLC.Cytoprotective activity assay demonstrated that both Tat-GluA2-3Y and CMT-C3Y exhibited activity at concentrations above 125 nmol/L,with CMT-C3Y showing superior activity as compared to Tat-GluA2-3Y.The results of the transmembrane assay demonstrated that,compared to Tat-GluA2-3Y,CMT-C3Y exhibited significant transmembrane capabilities at all tested concentrations(P<0.001).CMT-C3Y was classified as a highly permeable compound,whereas Tat-GluA2-3Y was categorized as a moderately permeable compound.Plasma stability studies indicated that over 50%of Tat-GluA2-3Y was metabolized after 4 h of co-incubation with rat plasma.After 8 h of coincubation with CMT-C3Y,the remaining amount was 88.1%,and no obvious degradation phenomenon occurred.In human liver microsomal stability tests,the half-life of Tat-GluA2-3Y was 26.1 min,as compared to 103.8 min for CMT-C3Y,highlighting the enhanced stability of CMT-C3Y.Tat-GluA2-3Y and CMT-C3Y were classified as a fast-metabolizing drug and a moderate-metabolizing drug,respectively.Animal experiments further demonstrated that at a dose of 8 mg/kg the neuroprotective activity of CMT-C3Y was significantly superior to that of Tat-GluA2-3Y(P<0.001).Conclusion The designed bicyclic peptide CMT-C3Y demonstrates significantly higher cell-penetrating efficiency and superior plasma stability as compared to Tat-GluA2-3Y,along with enhanced neuroprotective activity at both cellular and animal levels.

Objective:To discuss the damage effect of vesicular stomatitis virus(VSV)on the vascular endothelial(VE)barrier,and to clarify its mechanism.Methods:The canine kidney cells were used to amplify VSV.The half tissue culture infective dose(TCID50)of VSV was determined using mouse brain endothelial tumor bEnd.3 cells,and subsequent experiment was conducted using 300 times the TCID50.The bEnd.3 cells were divided into infection 0 h group,infection 4 h group,infection 8 h group,and infection 12 h group for VE barrier damage experiments due to VSV infection.The bEnd.3 cells were also divided into control group,infection group,and correction group for experiments to inhibit the VSV replication and restore the VE barrier.The bEnd.3 cells were inoculated into Transwell chambers to construct an in vitro VE barrier model.Cell voltage resistance meter was used to detect the transepithelial resistance(TER)in various groups after the bEnd.3 cells were infected with VSV at different time points;fluorescein isothiocyanate-dextran leakage assay was used to detect the permeability coefficients of the cells in various groups;immunofluorescence staining was used to observe the localization changes of VE-cadherin,β-catenin,and phosphorylated β-catenin(p-β-catenin)in cytoskeleton and adherens junctions(AJs)of the bEnd.3 cells after VSV infection;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Wnt and β-catenin mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of Wnt,β-catenin,and p-β-catenin proteins in the cells in various groups.Results:The TCID50 of VSV was 10-4.5·100 μL-1.TheTranswell chamber experiment results showed that compared with infection 0 h group,the TERs in the cells in the other groups were significantly decreased(P＜0.05),and the permeability coefficients were significantly increased(P＜0.05).The immunofluorescence staining results showed that compared with control group,the cytoskeleton of the bEnd.3 cells in infection group was disordered,the cell gaps was increased,the linear index of AJs was significantly decreased(P＜0.05),and β-catenin and p-β-catenin translocated from the cell membrane to the perinuclear area.The RT-qPCR results showed that compared with infection 0 h group,the expression levels of Wnt mRNA in the cells in the other groups were significantly decreased(P＜0.05),while the expression levels of β-catenin mRNA showed no statistically significant difference(P＞0.05).The Western blotting results showed that compared with infection 0 h group,the expression levels of Wnt protein in the cells in the other groups were significantly decreased(P＜0.05),the expression levels of β-catenin showed no statistically significant differences(P＞0.05),and the expression levels of p-β-catenin were significantly increased(P＜0.05).After inhibiting the VSV replication and correcting the low density lipoprotein receptor(LDLR)abnormalities,the Transwell chamber experiment results showed that compared with infection group,the TER in the cells in correction group was significantly increased(P＜0.05),and the permeability coefficient was significantly decreased(P＜0.05).The immunofluorescence staining results showed that compared with infection group,the gaps in the cells in correction group were reduced,and the perinuclear aggregation of β-catenin and p-β-catenin in the cells was restrained.The RT-qPCR results showed that compared with infection group,the expression level of Wnt mRNA in the cells in correction group was significantly increased(P＜0.05).The Western blotting results showed that compared with infection group,the expression level of Wnt protein in the cells in correction group was significantly increased(P＜0.05),the expression level of β-catenin showed no statistically significant difference(P＞0.05),and the expression level of p-P-catenin was significantly decreased(P＜0.05).Conclusion:VSV infection can cause the LDLR inactivation,reduce the expression level of Wnt protein,increase the phosphorylation level of β-catenin and cause its internalization,disrupt the stability of AJs,and ultimately lead to VE barrier damage.

Tetrandrine(TET),a natural bisbenzyl isoquinoline alkaloid extracted from Stephania tetrandra S.Moore,has diverse pharmacological effects.However,its effects on melanoma remain unclear.Cellular prolif-eration assays,multi-omics analyses,and xenograft models were used to determine the effect of TET on melanoma.The direct target of TET was identified using biotin-TET pull-down liquid chromatograph-mass spectrometry(LC-MS),cellular thermal shift assays,and isothermal titration calorimetry(ITC)analysis.Our findings revealed that TET treatment induced robust cellular autophagy depending on activating transcription factor 6(ATF6)-mediated endoplasmic reticulum(ER)stress.Simultaneously,it hindered autophagic flux by inducing cytoskeletal protein depolymerization in melanoma cells.TET treatment resulted in excessive accumulation of reactive oxygen species(ROS)and simultaneously triggered mitophagy.Sirtuin 5(SIRT5)was ultimately found to be a direct target of TET.Mechanistically,TET led to the degradation of SIRT5 via the ubiquitin(Ub)-26S proteasome system.SIRT5 knockdown induced ROS accumulation,whereas SIRT5 overexpression attenuated the TET-induced ROS accumula-tion and autophagy.Importantly,TET exhibited anti-cancer effects in xenograft models depending on SIRT5 expression.This study highlights the potential of TET as an antimelanoma agent that targets SIRT5.These findings provide a promising avenue for the use of TET in melanoma treatment and underscore its potential as a therapeutic candidate.

In the face of DNA damage caused by various factors, cells have a set of response and repair mechanisms. Cell cycle arrest plays an important role in the DNA damage repair, which provides enough time for repairing damaged DNA. Research on cell cycle regulation focuses on cyclin-dependent protein kinases (CDKs) and cell cycle checkpoints. In the process of DNA damage repair, phosphatidylinositol-3-kinase like kinases (PIKKs) which are recruited to the DNA damage sites can activate cell cycle checkpoint-related proteins to halt cell cycle. In the common DNA damage repair pathways, such as base excision repair (BER), nucleotide excision repair (NER) , mismatch repair (MMR) , and DNA double-strand break repair, the recruitment of repair-related proteins also plays a role in the cell cycle regulation. In this paper, the relationship between the main forms of DNA damage repair and cell cycle arrest and relevant research progress were reviewed.

Objective To study the stability of anticoagulant peptide Hirulog-S and its lyophilized product, and to provide data on the storage conditions and clinical applications.Methods RP-HPLC was used to determine the content and the related substances of Hirulog-S and its lyophilized powder with influence factor test, accelerated test and long-term storage test.Results Light, temperature and humidity had no significant effect on the stability of Hirulog-S and its lyophilized powder in the influence factor test.The content and related substances of Hirulog-S and its lyophilized powder did not significantly change in the accelerated test ( 40℃, RH75%) and 24-month long-term storage test at room temperature and 4℃.Conclusion Hirulog-S and its lyophilized product are very stable, even after being stored at room temperature for two years.

Objective To design and synthesize a series of new type four hydrogen quinoline-benzyl/benzimidazole amine derivatives as a potential new inhibitor targeting auxiliary receptor CXCR 4, and determine their inhibitory activities to HIV-1.Methods Based on HIV-1 receptor CXCR4 inhibitors containing three nitrogen structure-activity motif and CCR5 partial hydrophobic pharmacophore , a series of new compounds were designed , synthesized and characterized by 1 HNMR and MS.The inhibitory activities of these compounds were determined using HIV-1 IIIB virus.Results and Conclusion Ten target compounds are synthesized .Four hydrogen quinoline-benzimidazole amine derivatives exhibit good anti-HIV activity(IC50 <1 μmol/L), but four hydrogen quinoline-benzyl amine compounds are less active ((IC50 >8 μmol/L).