OBJECTIVE To establish the UPLC fingerprint and the method for multi-component content determination in Jingangteng capsules， and to evaluate its quality by combining chemical pattern recognition analysis. METHODS An UPLC method was established. Separation was performed on a Zorbax SB-C 18 Rapid Resolution HD column， with acetonitrile-0.1% formic acid as the mobile phase for gradient elution.Using the Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicines （2012 edition）， UPLC fi ngerprints were established for 10 batches of Jingangteng capsules， and similarity was evaluated. SPSS 22.0 and SIMCA 14.1 software were used to perform hierarchial-cluster analysis and orthogonal partial least squares discriminant analysis （OPLS-DA）， respectively. The same UPLC method was employed to determine the contents of chlorogenic acid， 3，5-dihydroxy-2-methylbenzoic acid-3- O -glucoside （M1）， caffeic acid， astilbin， oxyresveratrol， quercitrin and resveratrol in the 10 batches of samples. RESULTS A total of 17 common peaks were identified in UPLC fingerprints of the 10 batches of samples， of which 7 were identified as chlorogenic acid， M1， caffeic acid， astilbin， oxyresveratrol， quercitrin， and resveratrol. The similarities of 10 batches of samples ranged from 0.820 to 0.985. The results of hierarchial-cluster analysis showed that 10 batches of samples were grouped into four categories： S1-S4 formed one group， S5 and S6 formed another， S7， S8 and S10 formed a third， and S9 formed a fourth， consistent with the OPLS-DA results； the variable importance projection values for peaks 7， 10， 2， 16 （resveratrol）， 13 （oxyresveratrol）， 11， 6 （caffeic acid）， 5 （M1） and 15 （quercitrin） were ＞1. Quantitative analysis results showed that the contents of chlorogenic acid， M1， caffeic acid， astilbin， oxyresveratrol， quercitrin， and resveratrol were 1.650 8-4.213 7， 0.636 2-2.161 7， 0.031 0-0.086 5， 0.239 1-1.069 3， 0.211 9-1.104 0， 0.488 8-2.399 2， and 0.164 0-0.699 8 mg/g， respectively. CONCLUSIONS UPLC fingerprint and content determination methods established in this study are simple to operate， accurate， reliable and reproducible； when combined with chemical pattern recognition analysis， they can be used to evaluate the quality of Jingangteng capsules. Nine components， such as resveratrol， oxyresveratrol， caffeic acid， M1 and quercitrin， may serve as markers of quality variation.

Objective:To evaluate the role of tubulin tyrosin ligase like-6 (TTLL6)-mediated microtubule polyglutamylation and Spastin(a microtubule cleaving protein)-induced excessive microtubule cleavage in the developmental impairment of dendritic spines in neonatal mice following repeated sevoflurane anesthesia, by utilizing TTLL6 conditional knockout mice.Methods:Fifty SPF female TTLL6 brain tissue-specific knockout (TTLL6 CKO: Camk2-Cre + ; TTLL6 f/f) and fifty control (TTLL6 CON: TTLL6 f/f) mice with C57BL/6J background, aged 6 days old were selected.TTLL6 CKO mice were divided into TTLL6 CON control group and TTLL6 CON sevoflurane group, and TTLL6 CKO mice were divided into TTLL6 CKO control group and TTLL6 CKO sevoflurane group, with 25 mice in each group by random block method.Mice in the sevoflurane groups were exposed to 3% sevoflurane with 60% O 2 for 2 hours daily on postnatal days 6, 8, and 10.The mice in control groups received only 60% O 2 under the same condition.The polyGlu-Tubulin and postsynaptic density 95(PSD95) protein expression were detected using Western blot. The expressions of TTLL6, Spastin, and α-Tubulin were assessed via immunofluorescence.Golgi staining and electron microscopy were employed to observe the density of hippocampal dendritic spines and synaptic conditions. The Morris water maze test was used to evaluate spatial memory capabilities. Statistical analysis was performed using GraphPad Prism 8.0 software. Results:(1) Behavioral results showed significant time and group interactions among the four groups in terms of latency to find the platform ( F=8.22, P<0.001).Mice in the TTLL6 CON sevoflurane group had a significantly longer escape latency on days 3-7 compared with the TTLL6 CON control group (all P<0.05), while there was no significant difference between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group (all P>0.05). The number of platform crossings differed significantly among the four groups ( H=11.95, P=0.007).The TTLL6 CON sevoflurane group had significantly fewer crossing times than the TTLL6 CON control group ( P<0.05), but no significant difference was observed between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group ( P>0.05). (2) Golgi staining and electron microscopy results revealed significant differences in dendritic spine density and synapse number among the four groups( F=29.00, 41.94, both P<0.001). The dendritic spine density ((5.83±0.40)/10 μm) and the number of synapses ((3.67±0.58)/10 μm) in the TTLL6 CON sevoflurane group were both significantly lower than those in the TTLL6 CON control group ((12.87±1.70)/10 μm, (9.33±0.57)/10 μm)(both P<0.05). In contrast, there was no statistically significant difference between the TTLL6 CKO sevoflurane group and TTLL6 CKO control group (both P>0.05). (3) Immunofluorescence results showed significant differences in the percentage of TTLL6 and Spastin and α-Tubulin co-expressed positive cells in the CA3 region of the hippocampus among the four groups of mice ( F=215.20, 26.08, both P<0.001). The percentage of TTLL6 and Spastin and α-Tubulin co-expressed positive cells in the TTLL6 CON sevoflurane group ((16.75±1.81) %, (47.98±8.42) %) were significantly higher than those in the TTLL6 CON control group ((2.44±0.58) %, (20.07±4.54) %)(both P<0.05), while there was no statistically significant difference between the TTLL6 CKO sevoflurane group and TTLL6 CKO control group ( P>0.05). (4) Western blot results indicated significant differences in the expression of polyGlu-Tubulin and PSD95 proteins in the hippocampal tissue among the four groups of mice ( F=19.66, 8.57, both P<0.001). The TTLL6 CON sevoflurane group had higher polyGlu-Tubulin expression (0.86±0.19) and lower PSD95 expression (0.61±0.13) compared to the TTLL6 CON control group (0.51±0.11, 1.01±0.07) (both P<0.05).However, there was no significant difference between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group ( P>0.05). Conclusion:The mechanism underlying long-term cognitive impairment in developing brain of neonatal mice caused by repeated sevoflurane anesthesia may relate to the upregulation of TTLL6-induced microtubule polyglutamylation and accelerated Spastin-mediated microtubule severing, which ultimately leads to abnormal dendritic spine development.

HDAC7, a member of class IIa HDACs, plays a pivotal regulatory role in tumor, immune, fibrosis, and angiogenesis, rendering it a potential therapeutic target. Nevertheless, due to the high similarity in the enzyme active sites of class IIa HDACs, inhibitors encounter challenges in discerning differences among them. Furthermore, the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes, leading to a limited impact of enzymatic inhibitors on their function. In this study, proteolysis targeting chimera (PROTAC) technology was employed to develop HDAC7 drugs. We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma (DLBCL) and acute myeloid leukemia (AML) cells. Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7, thereby exerting proliferative inhibition in DLBCL. Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies, particularly in DLBCL and AML.

OBJECTIVES: To explore the therapeutic mechanism of compound Centella asiatica formula (CCA) for alleviating Schistosoma japonicum (Sj)-induced liver fibrosis in mice. METHODS: The active components and targets of CCA were identified using the TCMSP database with cross-analysis of Sj-related liver fibrosis targets. A "drug-component-target-pathway-disease" network was constructed using Cytoscape 3.9.1. Functional enrichment analysis (GO/KEGG) was performed using DAVID. Molecular docking study was carried out to validate interactions between the core targets and the key compounds. For experimental validation of the results, 36 mice were divided into control group, Sj-infected model group, and CCA-treated groups. In the latter two groups, liver fibrosis was induced via abdominal infection with Sj cercariae for 8 weeks, followed by 8 weeks of daily treatment with CCA decoction or saline. Hepatic pathology of the mice was assessedwith HE and Masson staining, and hepatic expressions of collagen-I and collagen-III were detected using immunohistochemistry; serum IL-6 and TNF-α levels were determined with ELISA. Hepatic expressions of TLR4 and MyD88 proteins were analyzed with Western blotting. RESULTS: We identified a total of 107 bioactive CCA components and 791 targets, including 37 intersection targets linked to Sj-induced fibrosis. The core targets included TNF, TP53, JUN, MMP9, and CXCL8, involving the IL-17 signaling, lipid metabolism, TLR4/MyD88 axis, and cancer pathways. Molecular docking study confirmed strong binding affinity between quercetin (a primary CCA component) and TNF/TP53/JUN/MMP9. In Sj-infected mouse models, CCA treatment significantly attenuated hepatic inflammatory cell infiltration, reduced collagen-I and collagen-III deposition, improved tissue architecture, reduced serum IL-6 and TNF-α levels, and downregulated TLR4 and MyD88 expressions in the liver. CONCLUSIONS CCA mitigates Sj-induced liver fibrosis by targeting TNF, TP53, JUN, and MMP9 to modulate the TLR4/MyD88 pathway, thereby suppressing pro-inflammatory cytokine release, inhibiting hepatic stellate cell activation, reducing collagen deposition, and preventing granuloma formation in the liver.
Animals ; Toll-Like Receptor 4/metabolism* ; Mice ; Myeloid Differentiation Factor 88/metabolism* ; Schistosoma japonicum ; Liver Cirrhosis/parasitology* ; Schistosomiasis japonica ; Signal Transduction ; Molecular Docking Simulation ; Inflammation ; Centella/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Tumor Necrosis Factor-alpha/metabolism*

Objective:To analyze the effect of continuous positive airway pressure (CPAP) on maternal and neonatal outcomes in pregnant women with obstructive sleep apnea syndrome (OSAS), especially on the incidence of hypertensive disorder in pregnancy (HDP) in women with moderate to severe OSAS.Methods:A total of 180 pregnant women with OSAS who were diagnosed through sleep monitoring during pregnancy due to high-risk factors of OSAS and registered in Peking University People′s Hospital from January 2021 to May 2024 were selected as the study subjects. Clinical data were collected from medical records for retrospective analysis. According to whether they received standardized treatment with CPAP, they were divided into the CPAP treatment group (42 cases) and the control group (138 cases). The CPAP treatment group consisted of 9 pregnant women with moderate to severe OSAS, while the control group consisted of 34 pregnant women with moderate to severe OSAS. The maternal and neonatal outcomes, the incidence of HDP, placental weight after delivery and placental weight/neonatal birth weight ratio were compared between the two groups.Results:(1) The average gestational age of pregnant women in the CPAP treatment group was higher than that in the control group [(38.7±1.0) vs (38.0±1.4) weeks], the proportion of infants small for gestational age (SGA) in the CPAP treatment group was lower [0 (0/42) vs 12.3% (17/138)], and the birth weight of infants in the CPAP treatment group was bigger [(3 396±475) vs (3 082±710) g); the differences between the two groups were statistically significant (all P<0.05). There were no significant differences between the CPAP treatment group and the control group in terms of delivery mode, rates of postpartum hemorrhage and preterm birth, umbilical artery blood gas analysis pH<7.1, lactate≥6.0 mmol/L, base excess<-12.0 mmol/L and the incidence of gestational diabetes mellitus and HDP (all P>0.05). (2) The placental weight of the CPAP treatment group was significantly lower than that of the control group [(554.0±70.6) vs (615.7±119.1) g], the placental weight/newborn birth weight ratio of the CPAP treatment group was significantly lower than that of the control group (median: 0.17 vs 0.19), and the differences were statistically significant (all P<0.05). (3) The incidence of HDP in pregnant women with moderate to severe OSAS in the CPAP treatment group was lower than that in the control group [1/9 vs 61.8% (21/34)], and the difference was statistically significant ( P<0.05). Conclusions:CPAP treatment could prolong the gestational age in pregnant women with OSAS, reduce the incidence of SGA, increase the birth weight of infants, and reduce the incidence of HDP in pregnant women with moderate to severe OSAS, and is worth promoting in clinical practice. The improvement of neonatal outcomes by CPAP treatment is closely related to the placenta, which is worthy of further exploration.

Objective:It aims to examine the effects of the employee medical insurance outpatient mutual assistance on medical insurance costs for patients with diabetes mellitus,offering insights for optimizing outpatient insurance policies and chronic disease management strategies.Methods:Outpatient cost settlement data of urban employees with diabetes mellitus in Lanzhou from 2022 to 2023 was collected.Univariate analysis and interrupted time-series models were used to compare relevant medical insurance cost indicators before and after the reform.Results:The inpatient data of 765 730 diabetes mellitus patients were included in the study,and male patients account for 62.67％.After the reform,average per-visit pooling fund expenditure,average per-visit individual payment expenditures,average per-visit personal account expenditure,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments were decreased(P＜0.05).Moreover,average per-visit pooling fund expenditure,average per-visit individual account expenditure,average per-visit cash payment,average per-visit eligible expense amount,average per-visit total fund payment,and the average per-visit proportion of basic medical pooling payments showed a notable declining trend post-reform(P＜0.05).In contrast,the per-visit fully out-of-pocket expenditure exhibited no significant change before and after the reform(P＞0.05).Conclusion:The reform of the employee medical insurance outpatient mutual assistance system has alleviated the economic burden of disease for diabetic patients and improved the efficiency of medical insurance fund utilization,but it reduced the proportion of basic medical pooling payments.It is recommended to continuously refine the outpatient medical insurance payment system,strengthen supervision of medical expenses and service quality,and balance patient benefits with fund pressure to enhance chronic disease outpatient benefits.

RNA binding motif protein 8A(RBM8A)is an RNA binding protein,which is mainly in-volved in translation and cell cycle regulation.In addition,RBM8A is a core factor of the exon-junc-tion complex(EJC),which is highly expressed in cells,especially in cancer cells,and abnormally expressed in cytoplasm and nucleus.Studies have shown that RBM8A plays a key regulatory role in the replication process of some viruses,such as Flaviviridae viruses.Therefore,whether RBM8A is involved in the replication of pseudorabies virus(PRV)is unknown.Therefore,this study proved whether RBM8A is involved in the replication of PRV.In order to study the effect of RBM8A pro-tein on PRV replication,the eukaryotic expression plasmid pCAGGS-HA-RBM8A was designed and constructed to express RBM8A,and sh-RBM8A was simultaneously designed and constructed to overexpress and inhibit RBM8A.qRT-PCR and Western blot were used to detect the effect of RBM8A on PRV replication.At the same time,PRV-GB standard plasmid was constructed to make PRV proliferation standard curve.After overexpression and inhibition of RBM8A,DNA was ex-tracted.Virus copy number was calculated by qRT-PCR to further detect the effect of RBM8A on PRV replication.The results showed that overexpression of RBM8A inhibited PRV replication and decreased the copy number of the virus,while overexpression of shRBM8A promoted PRV replication and increased the copy number of the virus.This study shows that RBM8A can inhibit PRV replication,which provides reference for the functional study of RBM8A and lays a founda-tion for the mechanism of anti-PRV replication.

Objective This study aimed to investigate the value of PET/CT radiomics combined with machine learning in differentia-ting primary central nervous system lymphoma(PCNSL)from brain metastases(BM).Methods Sixty-nine patients with 127 lesions(including 43 PCNSL lesions and 84 BM lesions)who attended the Department of Nuclear Medicine of Lanzhou University Second Hospi-tal from January 2019 to November 2024 were selected and divided into a training set(n=88)and a validation set(n=39)in a 7∶3 rati-o.Radiomics features of PET and CT were extracted using the 3D slicer,and feature dimensionality reduction was performed using least absolute shrinkage and selection operator(LASSO)regression combined with 5-fold cross-validation to construct three classical machine learning models-Logistic regression(LR),decision tree(DT),and support vector machine(SVM),and the diagnostic efficacy of each model was evaluated using receiver operating characteristic(ROC)curves and confusion matrices.Results The SVM model based on PET imaging performed best in the validation set[area under the curve(AUC)=0.917,sensitivity=84.6％,specificity=92.3％]and was significantly better than the CT model(AUC=0.787,sensitivity=73.1％,specificity=76.9％).Conclusion The support vector machine model constructed based on PET radiomics metabolic features demonstrated high diagnostic value in the preoperative differentia-tion between PCNSL and BM,showing potential for clinical application as a non-invasive auxiliary diagnostic tool.

Objective To explore the mechanism of GS-9620 in improving imiquimod(IMQ)-induced psoriasis-like inflammation by regulating the Th1/Th17-related immune response,and to investigate its regulatory effect on the gut microbiota in mice.Methods An IMQ-induced psoriasis-like inflammation model was established in BALB/c mice.The severity of the skin lesions was evaluated by psoriasis area and severity index(PASI)score.The proportions of CD4+interleukin(IL)-17+and CD4+interferon(IFN)-γ+cells in spleen tissue were detected by flow cytometry.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,IL-1β,and IL-6 in skin tissues were determined by enzyme-linked immunosorbent assay,and pathological analysis was performed by hematoxylin/eosin staining.The effects of GS-9620 on the structure of the gut microbiota in control,IMQ model,and GS-9620-treated mice were detected by 16S rRNA sequencing.Results GS-9620 significantly reduced the PASI score in IMQ-induced mice and effectively reduced the proportions of CD4+IL-17+and CD4+IFN-γ+cells in the spleen.GS-9620 also significantly down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in skin tissues.16S rRNA sequencing showed that GS-9620 significantly regulated the abundance of gut microbiota related to inflammation,including the relative abundances of bacteria such as Lachnospiraceae_NK4A136_group,Lachnospiraceae_UCG-008,Alloprevotella,Desulfovibrio,Prevotellaceae_UCG-001,and Alistipes.Conclusions GS-9620 effectively alleviates IMQ-induced psoriasis-like skin inflammation in mice by regulating the expression of Th1/Th17-related inflammatory factors.It may also improve IMQ-induced clinical symptoms by regulating the structure of the gut microbiota,thus providing a new theoretical basis for the treatment of psoriasis.The result of this study provide important experimental evidence to support further investigations into the application of GS-9620 for the treatment of psoriasis.

Objective:To evaluate the role of tubulin tyrosin ligase like-6 (TTLL6)-mediated microtubule polyglutamylation and Spastin(a microtubule cleaving protein)-induced excessive microtubule cleavage in the developmental impairment of dendritic spines in neonatal mice following repeated sevoflurane anesthesia, by utilizing TTLL6 conditional knockout mice.Methods:Fifty SPF female TTLL6 brain tissue-specific knockout (TTLL6 CKO: Camk2-Cre + ; TTLL6 f/f) and fifty control (TTLL6 CON: TTLL6 f/f) mice with C57BL/6J background, aged 6 days old were selected.TTLL6 CKO mice were divided into TTLL6 CON control group and TTLL6 CON sevoflurane group, and TTLL6 CKO mice were divided into TTLL6 CKO control group and TTLL6 CKO sevoflurane group, with 25 mice in each group by random block method.Mice in the sevoflurane groups were exposed to 3% sevoflurane with 60% O 2 for 2 hours daily on postnatal days 6, 8, and 10.The mice in control groups received only 60% O 2 under the same condition.The polyGlu-Tubulin and postsynaptic density 95(PSD95) protein expression were detected using Western blot. The expressions of TTLL6, Spastin, and α-Tubulin were assessed via immunofluorescence.Golgi staining and electron microscopy were employed to observe the density of hippocampal dendritic spines and synaptic conditions. The Morris water maze test was used to evaluate spatial memory capabilities. Statistical analysis was performed using GraphPad Prism 8.0 software. Results:(1) Behavioral results showed significant time and group interactions among the four groups in terms of latency to find the platform ( F=8.22, P<0.001).Mice in the TTLL6 CON sevoflurane group had a significantly longer escape latency on days 3-7 compared with the TTLL6 CON control group (all P<0.05), while there was no significant difference between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group (all P>0.05). The number of platform crossings differed significantly among the four groups ( H=11.95, P=0.007).The TTLL6 CON sevoflurane group had significantly fewer crossing times than the TTLL6 CON control group ( P<0.05), but no significant difference was observed between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group ( P>0.05). (2) Golgi staining and electron microscopy results revealed significant differences in dendritic spine density and synapse number among the four groups( F=29.00, 41.94, both P<0.001). The dendritic spine density ((5.83±0.40)/10 μm) and the number of synapses ((3.67±0.58)/10 μm) in the TTLL6 CON sevoflurane group were both significantly lower than those in the TTLL6 CON control group ((12.87±1.70)/10 μm, (9.33±0.57)/10 μm)(both P<0.05). In contrast, there was no statistically significant difference between the TTLL6 CKO sevoflurane group and TTLL6 CKO control group (both P>0.05). (3) Immunofluorescence results showed significant differences in the percentage of TTLL6 and Spastin and α-Tubulin co-expressed positive cells in the CA3 region of the hippocampus among the four groups of mice ( F=215.20, 26.08, both P<0.001). The percentage of TTLL6 and Spastin and α-Tubulin co-expressed positive cells in the TTLL6 CON sevoflurane group ((16.75±1.81) %, (47.98±8.42) %) were significantly higher than those in the TTLL6 CON control group ((2.44±0.58) %, (20.07±4.54) %)(both P<0.05), while there was no statistically significant difference between the TTLL6 CKO sevoflurane group and TTLL6 CKO control group ( P>0.05). (4) Western blot results indicated significant differences in the expression of polyGlu-Tubulin and PSD95 proteins in the hippocampal tissue among the four groups of mice ( F=19.66, 8.57, both P<0.001). The TTLL6 CON sevoflurane group had higher polyGlu-Tubulin expression (0.86±0.19) and lower PSD95 expression (0.61±0.13) compared to the TTLL6 CON control group (0.51±0.11, 1.01±0.07) (both P<0.05).However, there was no significant difference between the TTLL6 CKO sevoflurane group and the TTLL6 CKO control group ( P>0.05). Conclusion:The mechanism underlying long-term cognitive impairment in developing brain of neonatal mice caused by repeated sevoflurane anesthesia may relate to the upregulation of TTLL6-induced microtubule polyglutamylation and accelerated Spastin-mediated microtubule severing, which ultimately leads to abnormal dendritic spine development.