ObjectiveTo systematically identify the chemical constituents of Xuefu Zhuyutang（XFZY） and quantitatively determine its main components， aiming to elucidate its pharmacodynamic material basis and provide a scientific foundation for improving its quality control standards. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS） was employed for qualitative analysis of XFZY， and the identification of compounds was accomplished by comparing their retention times， secondary MS fragment ion information， 52 reference standards and relevant databases， followed by attribution of their herbal sources. A total of 22 representative compounds were screened out， and UPLC-quadrupole-linear ion trap mass spectrometry（UPLC-Q-TRAP-MS/MS） was applied for quantitative analysis of the compounds in the formula. ResultsA total of 77 compounds were identified in XFZY， including 31 flavonoids mainly derived from Aurantii Fructus， Glycyrrhizae Radix et Rhizoma， Persicae Semen， Carthami Flos， Bupleuri Radix， Paeoniae Radix Rubra and Achyranthis Bidentatae Radix， 24 terpenoids mainly derived from Platycodonis Radix， Glycyrrhizae Radix et Rhizoma， Paeoniae Radix Rubra and Rehmanniae Radix， 9 phenylpropanoids and their derivatives mainly derived from Chuanxiong Rhizoma， Angelicae Sinensis Radix and Rehmanniae Radix， 4 phenolic acids mainly derived from Chuanxiong Rhizoma， Angelicae Sinensis Radix and Paeoniae Radix Rubra， 3 saccharides mainly derived from Rehmanniae Radix and Achyranthis Bidentatae Radix， and 6 other compounds mainly derived from Persicae Semen， Rehmanniae Radix and Angelicae Sinensis Radix. The results of quantitative analysis showed that the contents of protocatechuic acid， hydroxypaeoniflorin， amygdalin， vanillic acid， paeoniflorin， liquiritin apioside， liquiritin， isoquercitrin， naringin， cosmosiin， hesperidin， neohesperidin， isoliquiritin， liquiritigenin， naringenin， benzoylpaeoniflorin， hesperetin， isoliquiritigenin， formononetin， glycyrrhizic acid， nobiletin and ligustilide in XFZY were determined to be 0.12， 1.57， 54.53， 0.29， 36.17， 4.29， 4.84， 0.09， 46.67， 0.04， 3.44， 31.95， 0.82， 0.10， 0.11， 0.43， 0.07， 0.03， 0.01， 8.24， 0.13， 1.81 mg·g^-1. ConclusionThe qualitative method established in this study enables rapid and sensitive analysis of the chemical constituents in XFZY. Among the identified compounds， 52 are confirmed by reference standards， ensuring the accuracy of identification. The quantitative analysis of 22 key components provides a reliable experimental basis for the pharmacodynamic material basis research and quality control standard improvement of XFZY.

Hepatocellular carcinoma （HCC）， the predominant subtype of primary liver cancer， ranks among the top in both incidence and mortality rates of malignant tumors in China. In its early stages， the disease may present with subtle or nonspecific symptoms， often leading to poor clinical prognosis and low patient survival rates， which makes it a significant public health concern. The pathogenesis is associated with multiple factors， including hepatitis virus infection， alcohol consumption， obesity， drug-induced liver injury， and immune disorders， which may interact synergistically to promote disease development. Currently， mainstream therapeutic approaches for HCC in modern medicine encompass surgical resection， liver transplantation， radiofrequency ablation， radiotherapy， and chemotherapy， but they all have certain limitations， such as large side effects and poor prognosis， imposing substantial psychological distress and financial strain on affected individuals. With a rich historical background in hepatic malignancy management， traditional Chinese medicine offers therapeutic benefits characterized by multi-targeted mechanisms， multi-level regulation， minimal adverse effects， and reduced likelihood of disease recurrence. It can not only enhance the curative effect， but also reduce the side effects of radiotherapy， chemotherapy， and surgery. Thus， it has attracted widespread attention. Extensive research has demonstrated that traditional Chinese medicine exhibits significant antitumor properties， along with notable anti-inflammatory and oxidative stress-reducing capabilities， and its mechanism may be related to the regulation of nuclear factor-kappa B （NF-κB） signaling pathway， which can affect multiple stages of hepatocarcinogenesis， such as cell proliferation， invasion， metastasis， and apoptosis. The mechanism of NF-κB signaling pathway in traditional Chinese medicine for HCC treatment has emerged as one of the pivotal research directions in current oncology studies. Based on the existing research foundation， a systematic literature review method was adopted to retrieve and analyze relevant Chinese and English literature in recent years. Integrating the molecular regulatory mechanisms of the NF-κB signaling pathway and its pivotal role in HCC pathogenesis and progression helped further explore the latest research advances in traditional Chinese medicine interventions targeting this pathway for HCC treatment. This approach may provide novel theoretical foundations and translational strategies for the prevention and management of HCC using traditional Chinese medicine.

Objective:To compare the performance of next-generation sequencing (NGS) and Sanger sequencing in investigating somatic hypermutation (SHM) status of immunoglobulin heavy chain variable region (IGHV) genes. It specifically focuses on identifying key factors contributing to discrepancies between the two methods, particularly under complex clonal backgrounds, to inform optimized strategies for clinical application.Methods:This retrospective analysis included 53 samples, comprising 43 identified as non-monoclonal and 10 as monoclonal using Sanger sequencing. All samples were further analyzed using NGS to assess IGHV SHM. The two methods were used for systematic comparison. For discordant cases, in-depth attribution analysis was conducted, considering factors, including clonal abundance quantification, differences in primer design, and interpretation criteria.Results:Among the 53 patients who underwent both Sanger and NGS testing, 36 were male and 17 were female, with a median age of 64 years (range: 33–88). Diagnoses included chronic lymphocytic leukemia (CLL) in 35 (66.0% ), diffuse large B-cell lymphoma in 9 (17.0% ), follicular lymphoma in 3 (5.7% ), mantle cell lymphoma in 3 (5.7% ), and other types in 3 (5.7% ) cases. In the 43 cases with non-monoclonal profiles using Sanger sequencing, NGS revealed 23 cases as biclonal or polyclonal, 17 as monoclonal, and 3 with no detectable clonality. The primary discrepancies between the two methods involved variations in clonality assessment, IGHV gene rearrangement types, and mutation rates. Among the 10 cases identified as monoclonal using Sanger sequencing, NGS detected biclonality and markedly different IGHV rearrangement types in 2 and 4 cases, respectively. Minor differences were observed in SHM percentage between the two methods; however, these did not substantially affect the overall determination of mutational status.Conclusion:Compared with Sanger sequencing, NGS exhibits superior performance in assessing IGHV SHM status under complex clonal conditions. It provides greater sensitivity and accuracy in detecting subclonal components and quantifying clonal proportions, thereby providing a more precise molecular basis for diagnosing and prognostically assessing lymphoid malignancies, including CLL.

ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal （HPG） axis by modified Erxian decoction in rats with perimenopausal syndrome （PMS） and to further analyze the expression of proteins related to the silent information regulator 1 （SIRT1）/hypothalamic kisspeptin （Kisspeptin）/gonadotropin-releasing hormone （GnRH） signaling pathway in the arcuate nucleus region （ARC） of the hypothalamus， so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method， with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling， including a sham operation group， a model group， an estradiol valerate group （0.09 mg∙kg^-1∙d^-1）， high-， medium-， and low-dose modified Erxian decoction groups （7.614， 3.807，1.903 5 g∙kg^-1∙d^-1）， with 8 rats in each group. The estradiol valerate group and the high-， medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days， and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the levels of estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and gonadotropin-releasing hormone （GnRH）. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to determine the expression levels of SIRT1， Kisspeptin， kisspeptin receptor （GPR54）， and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor （GnRH-R） in pituitary. ResultsCompared with the sham operation group， rats in the model group had a significantly increased body weight （P0.01）， reduced wet weight and index of uterus （P0.01）， endometrial thinning or atrophy， glandular atrophy， and a decreasing number of glands. Additionally， serum levels of E₂ and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased （P0.01）. Serum levels of FSH， LH， and GnRH， the expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus， and GnRH-R in pituitary significantly increased （P0.01）. Compared with the model group， the estradiol valerate group and the high-， medium-dose modified Erxian decoction groups had significantly reduced body weight， serum levels of FSH， LH， and GnRH， and expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary （P0.05， P0.01） and significantly increased wet weight and index of uterus， serum level of E₂， and expression of SIRT1 in the ARC region of the hypothalamus （P0.05， P0.01）. In addition， they showed thickened endometrium， increased number of endometrial glands， and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets， and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus， the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway， the regulation of the expression of GnRH-R in the pituitary， the restoration of secretion balance of gonadotropins， and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.

Objective:To compare the performance of next-generation sequencing (NGS) and Sanger sequencing in investigating somatic hypermutation (SHM) status of immunoglobulin heavy chain variable region (IGHV) genes. It specifically focuses on identifying key factors contributing to discrepancies between the two methods, particularly under complex clonal backgrounds, to inform optimized strategies for clinical application.Methods:This retrospective analysis included 53 samples, comprising 43 identified as non-monoclonal and 10 as monoclonal using Sanger sequencing. All samples were further analyzed using NGS to assess IGHV SHM. The two methods were used for systematic comparison. For discordant cases, in-depth attribution analysis was conducted, considering factors, including clonal abundance quantification, differences in primer design, and interpretation criteria.Results:Among the 53 patients who underwent both Sanger and NGS testing, 36 were male and 17 were female, with a median age of 64 years (range: 33–88). Diagnoses included chronic lymphocytic leukemia (CLL) in 35 (66.0% ), diffuse large B-cell lymphoma in 9 (17.0% ), follicular lymphoma in 3 (5.7% ), mantle cell lymphoma in 3 (5.7% ), and other types in 3 (5.7% ) cases. In the 43 cases with non-monoclonal profiles using Sanger sequencing, NGS revealed 23 cases as biclonal or polyclonal, 17 as monoclonal, and 3 with no detectable clonality. The primary discrepancies between the two methods involved variations in clonality assessment, IGHV gene rearrangement types, and mutation rates. Among the 10 cases identified as monoclonal using Sanger sequencing, NGS detected biclonality and markedly different IGHV rearrangement types in 2 and 4 cases, respectively. Minor differences were observed in SHM percentage between the two methods; however, these did not substantially affect the overall determination of mutational status.Conclusion:Compared with Sanger sequencing, NGS exhibits superior performance in assessing IGHV SHM status under complex clonal conditions. It provides greater sensitivity and accuracy in detecting subclonal components and quantifying clonal proportions, thereby providing a more precise molecular basis for diagnosing and prognostically assessing lymphoid malignancies, including CLL.

The seat of human intelligence is the human cerebral cortex, which is responsible for our exceptional cognitive abilities. Identifying principles that lead to the development of the large-sized human cerebral cortex will shed light on what makes the human brain and species so special. The remarkable increase in the number of human cortical pyramidal neurons and the size of the human cerebral cortex is mainly because human cortical radial glial cells, primary neural stem cells in the cortex, generate cortical pyramidal neurons for more than 130 days, whereas the same process takes only about 7 days in mice. The molecular mechanisms underlying this difference are largely unknown. Here, we found that bone morphogenic protein 7 (BMP7) is expressed by increasing the number of cortical radial glial cells during mammalian evolution (mouse, ferret, monkey, and human). BMP7 expression in cortical radial glial cells promotes neurogenesis, inhibits gliogenesis, and thereby increases the length of the neurogenic period, whereas Sonic Hedgehog (SHH) signaling promotes cortical gliogenesis. We demonstrate that BMP7 signaling and SHH signaling mutually inhibit each other through regulation of GLI3 repressor formation. We propose that BMP7 drives the evolutionary expansion of the mammalian cortex by increasing the length of the neurogenic period.
Animals ; Mice ; Humans ; Ependymoglial Cells/metabolism* ; Hedgehog Proteins/metabolism* ; Ferrets/metabolism* ; Cerebral Cortex ; Neurogenesis ; Mammals/metabolism* ; Neuroglia/metabolism* ; Bone Morphogenetic Protein 7/metabolism*

Gastric cancer is one of the most com-mon malignant tumors in the digestive system,which often occurs in middle-aged and elderly peo-ple.Traditional Chinese medicine recognizes gastric cancer as a kind of tumor characterized by fluid de-ficiency,heat accumulation and the growing bind-ing of toxins in the stomach.It is commonly treated with heat-clearing and detoxifying drugs in clinical practice.Hedyotis diffusa-Scutellaria barbata herb pair(HS)has the effects of clearing heat and detoxi-fying,promoting blood circulation,resolving car-buncle and expulsing boil,anti-inflammatory and analgesic,which are consistent with the etiology and pathogenesis of gastric cancer,therefore,it can be used for the treatment of gastric cancer.Modern pharmacological researches have con-firmed that HS can play an anti-gastric cancer role by inducing cell apoptosis,inhibiting cell prolifera-tion,inhibiting angiogenesis,improving immune mi-croenvironment and down-regulating telomerase activity.Herein,this review summarizes the active ingredients and related mechanism responsible for the anti-gastric cancer effect of HS,which will pro-vide the theoretical basis for its clinical use and the development of new drugs against gastric cancers.

Objective:To explore the relationship between tumour psychological trait, negative emotions, and psychosomatic status in patients with advanced lung cancer.Methods:A total of 261 patients with advanced lung cancer from January 2014 to March 2019 at the First Affiliated Hospital of Guangdong Pharmaceutical University and the Affiliated Cancer Hospital and Institute of Guangzhou Medical University were selected as the research subjects. General information questionnaire, tumour psychological scale(TPS), hospital anxiety and depression scale(HADS), and psychosomatic status scale for cancer patients(PSSCP)were utilized for cross-sectional investigation. Descriptive analysis and correlation analysis were performed using SPSS 26.0 statistical software. Structural equation modeling was conducted using AMOS 24.0, and the mediating effect was tested using the Bootstrap method.Results:The total score of TPS was 77.16±8.68. The total score of HADS was 16.67±3.81, and the sub-scale scores of anxiety and depression were 7.00±2.26 and 9.67±2.56.The total score of PSSCP was 37.09±4.99.Negative emotions were positively correlated with tumour psychological trait and psychosomatic status ( r=0.308, 0.390, both P<0.01), and tumour psychological trait was positively correlated with psychosomatic status ( r=0.478, P<0.01). The structural equation model demonstrated that negative emotions partially mediated the relationship between tumour psychological trait and psychosomatic status, accounting for 24.9%(0.217/0.870). Conclusion:The psychosomatic status is affected both directly by tumour psychological trait and indirectly through negative emotions.

Purpose To assess the feasibility of nuclear score combined with ultrasound features on indeterminate thyroid nodules in fine needle aspiration cytological diagnosis.Methods A consecutive cohort of 119 thyroid fine-needle aspiration(FNA)specimens with indeterminate diagnosis(TBSRTC cate-gory Ⅲ-Ⅴ)and available histopathologic follow-up was evalua-ted.Original sonographic images and cytological slides of each nodule were reviewed by two experienced sonographers and two experienced cytopathologists respectively,and evaluated jointly to obtain consensus.The identification of the optimal cut-off points of simplified nuclear score and ultrasound features score for the diagnosis of malignancy or low-risk neoplasm were evalu-ated using the receiver operating characteristic(ROC)curves and the assessment of the area under the ROC curve(AUC).The specificity,sensitivity,positive predictive value(PPV)and negative predictive value(NPV)of nuclear score and US score were evaluated from crosstabs based on Cut-off points and signif-icance were calculated.The diagnostic accuracy of simplified nuclear score combined with ultrasound features was estimated by Logistic regression and ROC curve analysis.Results Nucle-ar grooves,intra-nuclear inclusions and chromatin clearing were more common in malignancy/low-risk neoplasms(P = 0.001,0.013 and 0.001 respectively).A Cut-off point of≥2 for the simplified nuclear score was sensitive for defining malignancy/low-risk neoplasm,and the sensitivity was 99.1%.Nodule height greater than width,absent halo sign,irregular margin and absent intranodular vascularization were more common in malig-nancy/low-risk neoplasms(P = 0.003,0.007,0.003 and 0.004 respectively).When the Cut-off point was set as≥5,the PPV and specificity for defining malignancy/low-risk neoplasm were 95.9%and 78.6%respectively.ROC curve analysis showed that the diagnostic accuracy of nuclear score combined with US score(0.873,95%CI = 0.777-0.970)was higher than that of nuclear score alone(0.817,95%CI = 0.687-0.948,P =0.092)or US score alone(0.758,95%CI 0.637-0.879,P =0.047).But no significant difference was detec-ted between the combined score and the nuclear score.Conclu-sion The addition of simplified nuclear score and US score to FNA cytology can increase the diagnostic accuracy of cytology in indeterminate categories.

Glioblastoma (GBM) is the most common and aggressive malignant brain tumor in adults and is poorly controlled. Previous studies have shown that both macrophages and angiogenesis play significant roles in GBM progression, and co-targeting of CSF1R and VEGFR is likely to be an effective strategy for GBM treatment. Therefore, this study developed a novel and selective inhibitor of CSF1R and VEGFR, SYHA1813, possessing potent antitumor activity against GBM. SYHA1813 inhibited VEGFR and CSF1R kinase activities with high potency and selectivity and thus blocked the cell viability of HUVECs and macrophages and exhibited anti-angiogenetic effects both in vitro and in vivo. SYHA1813 also displayed potent in vivo antitumor activity against GBM in immune-competent and immune-deficient mouse models, including temozolomide (TMZ) insensitive tumors. Notably, SYHA1813 could penetrate the blood-brain barrier (BBB) and prolong the survival time of mice bearing intracranial GBM xenografts. Moreover, SYHA1813 treatment resulted in a synergistic antitumor efficacy in combination with the PD-1 antibody. As a clinical proof of concept, SYHA1813 achieved confirmed responses in patients with recurrent GBM in an ongoing first-in-human phase I trial. The data of this study support the rationale for an ongoing phase I clinical study (ChiCTR2100045380).