Hereditary spherocytosis (HS) is the most common inherited chronic hemolytic anemia and results from defects in proteins of the erythrocyte membrane skeleton. Nonsense mutations in ANK1 are a common cause of HS, yet current treatments remain largely supportive without addressing genetic defects. In this study, we identified two ANK1 nonsense mutations (p.R281X and p.Q744X) in unrelated HS pedigrees and generated K562 erythroid differentiation models using cytosine base editor-mediated knock-in to evaluate potential targeted therapies. Functional analyses confirmed that both mutations markedly reduced ankyrin-1 expression and disrupted membrane-skeletal integrity. We then assessed two targeted therapeutic strategies: translational readthrough-inducing drugs (TRIDs) and adenine base editors (ABEs). Gentamicin promoted translational readthrough at both mutant sites, whereas CC-90009 showed activity primarily at p.Q744X. Both agents partially restored full-length ankyrin-1 expression, accompanied by restored membrane stability. In parallel, ABE8e-mediated correction achieved efficient (>85%) and precise genomic repair in bulk-edited K562 populations, leading to near-complete recovery of ankyrin-1 expression and membrane-skeletal integrity with minimal off-target effects. Together, these findings provide exploratory proof-of-concept for targeted molecular intervention in ANK1 nonsense mutation-associated HS, highlighting TRIDs and ABEs as mechanistically distinct strategies with complementary strengths and limitations and suggesting their relative utility may depend on mutation-specific and clinical contexts.

Histone lactylation, a recently identified post-translational modification, is closely linked to the pathogenesis and progression of malignant tumors, making it a promising therapeutic target for acute myeloid leukemia (AML). Increasing clinical and experimental evidence indicates that elevated serum lactate dehydrogenase (LDH) levels serve as both a diagnostic biomarker and an indicator of poor prognosis in AML, reflecting enhanced glycolytic activity and chemotherapy resistance. Patrinia scabiosaefolia Fisch (PS), a traditional medicinal herb, exhibits broad pharmacological activities, including heat-clearing, detoxifying, antibacterial, antiviral, and antitumor effects. However, the molecular mechanisms underlying its antileukemic activity, particularly in AML, remain insufficiently defined. To elucidate how DEPS exerts therapeutic effects in AML through the HIF-1α-histone lactylation axis, we systematically established the association between DEPS-mediated inhibition of histone lactylation and cellular hypoxia. Lactylation pan-antibody incubation confirmed preliminarily that DEPS significantly inhibits histone lactylation modification. Lactate restoration experiments further demonstrated that DEPS markedly suppresses lactate-induced activation of HIF-1α signaling, reducing downstream metabolic proteins and VEGFA expression. Under hypoxic conditions, AML cells displayed increased proliferation and histone lactylation, both of which were attenuated by DEPS treatment. Sodium L-lactate enhanced the expression of HIF-1α and its downstream effectors (HK2, PDK1, PKM2, GLUT1, LDHA, and VEGFA), whereas subsequent DEPS exposure significantly reversed these changes. Ultimately, DEPS inhibits H3/H4 lysine lactylation, induces G2/M cell-cycle arrest, and promotes apoptosis in THP-1 and HL-60 cells, including doxorubicin-resistant HL-60 cells. Collectively, our findings reveal previously uncharacterized antileukemic mechanisms of DEPS involving suppression of the HIF-1α signaling pathway and histone lactylation. These results highlight the importance of epigenetic regulation in AML and support the therapeutic potential of DEPS in AML treatment and reversal of chemoresistance.

Currently, the American College of Medical Genetics and Genomics (ACMG) carrier screening (CS) guidelines do not recommend routinely screening for conditions with carrier frequencies ≤ 1:200. Advances in DNA sequencing and declining costs, alongside growing awareness, make it increasingly feasible to expand CS to cover a broader spectrum of conditions. We aimed to design a CS panel regardless of carrier frequency to evaluate its utility in less common disease. We developed a large CS panel through gene curation, phenotypic severity evaluation, independent of carrier frequency. A cohort of 2010 couples across 24 Chinese cities underwent couple-based simultaneous screening. Gene carrier rate (GCR) was calculated, with subsequent assessment of at-risk couple rate (ACR) across varying GCR thresholds. The results obtained from testing 1736 genes were presented. Among 2010 couples undergoing CS, 106 were identified as having an increased risk of offspring with at least one genetic condition. The initial ACR is 5.3

Objective:The present study aimed to investigate the association of the ATP-binding cassette subfamily B member 1 (ABCB1) 3435C/T and methylenetetrahydrofolate reductase (MTHFR) 677C/T and 1298A/C polymorphisms with susceptibility to RA and RA subtype. Methods:We enrolled 85 RA patients and 45 healthy individuals in this study. Genotyping for ABCB1 3435C/T, MTHFR 677C/T and 1298A/C was performed using fluorescence in situ hybridization technology (FISH). Results:No significant differences in the genotype and allele frequency distributions of ABCB1 3435C/T, MTHFR 1298A/C or MTHFR677C/T were detected between patients with RA and healthy individuals (p > 0.05). Logistic regression analysis, after adjusting for sex and age, showed that the MTHFR677C/T TT genotype was associated with increased susceptibility to RA compared with the CC genotype (TT versus CC, p = 0.034, OR = 2.316; 95%CI = 1.067-5.029); however, after Bonferroni correction, the association between the MTHFR677C/T TT genotype and RA susceptibility was no longer significant. That MTHFR 677C/T genotype frequency (CT versus CC, p = 0.040, OR = 6.504; 95% CI = 1.087-38.935) and dominant model (CT+TT versus CC, p = 0.025, OR = 6.556; 95% CI = 1.272-33.799) was associated with ACPA status among RA patients; and that MTHFR 677C/T genotype frequency (TT versus CC, p = 0.044, OR = 2.171; 95% CI = 1.020-4.623) and codominant model (CC versus CT versus TT, p = 0.026, OR = 2.059; 95% CI = 1.089-3.891) were associated with RF status among RA patients; Similarly, the associations observed in ACPA-positive and RF-positive subgroups also lost significance after Bonferroni correction. Conclusion:The MTHFR 677C/T polymorphism may be associated with susceptibility to RA and RA subtypes; ABCB1 3435C/T and MTHFR 1298A/C were not associated with the susceptibility to RA or RA subtypes.

Alzheimer's disease is a progressive syndrome characterized by cognitive dysfunction, with neuroinflammation and β-amyloid plaque deposition representing early pathological hallmarks. Activated microglia and astrocytes play pivotal roles in neuroinflammation, further exacerbating the progression of Alzheimer's disease. Calycosin, an active ingredient derived from radix astragali, binds to estrogen receptors to elicit estrogen-like effects and has demonstrated efficacy in alleviating cognitive impairment. However, the impact of calycosin on memory deficits and its underlying mechanisms in the very early stages of Alzheimer's disease remain unknown. In this study, we aimed to investigate whether calycosin can ameliorate early memory loss by inhibiting microglia and astrocyte activation in the initial stages of Alzheimer's disease. To this end, we selected 3-month-old APP/PS1 transgenic mice and administered 20 mg/kg of calycosin daily for 90 days. Our findings revealed that calycosin administration improved long-term memory impairment, but had no significant effect on short-term learning and memory. Furthermore, calycosin reduced the number of β-amyloid plaques and alleviated neuronal loss in the cortex, although no such effect was observed in the hippocampus. Notably, calycosin did not alter the number of activated astrocytes or microglia surrounding β-amyloid plaques. Collectively, these results suggest that the improvement in long-term memory function observed with calycosin is not mediated through the inhibition of glial cell activation. These findings contribute to our understanding of the potential mechanisms underlying the beneficial effects of calycosin in Alzheimer's disease and highlight the complexity of its actions in this disease.

Neuronal activity and sensory experience regulate the subunit stoichiometry of synaptic N-methyl-D-aspartate subtype glutamate receptors (NMDARs), a critical determinant for brain development, synaptic plasticity, and a line of neurological disorders. Here we found that Ras and Rab interactor 1 (RIN1), a neuron-specific protein in the brain, played an important role in dictating synaptic NMDAR subunit composition in spinal cord somatostatin-positive (SOM+) neuron, a key component in the spinal circuit transmitting mechanical pain in mice. Our data showed that the protein level of RIN1 was low early after birth, which progressively increased with synapse maturation and promoted the switch from synaptic GluN2B- to GluN2A-containing NMDARs. In adult mice, the nerve injury-induced pathological pain paralleled a significant increase of RIN1 protein in spinal SOM+ neurons, which drove a new round of GluN2B-to-GluN2A switching at mature synapses. Our data revealed the molecular mechanisms by which RIN1 differentially regulated the synaptic trafficking of GluN2B and GluN2A receptors, and implied that RIN1-mediated pathological switch of NMDAR subunit composition strikingly altered the analgesic efficacy of distinct NMDAR subunit antagonists with the development of neuropathic pain.

Cuproptosis is a newly identified form of cell death that relies on copper (Cu) ionophores to transport Cu into cancer cells. As a perennial herb, Patrinia scabiosaefolia Fisch (PS) has garnered significant attention owing to its analgesic, anti-inflammatory, antibacterial, and antitumor properties. Previous research has shown that the extract from PS (DEPS) can inhibit the growth of leukemia cell lines. However, the specific mechanism of its anti-leukemic effect has not been fully clarified. Therefore, this study was conducted to investigate the molecular mechanism of cuproptosis in the treatment of leukemia with DEPS. Our results demonstrated that DEPS up-regulated SLC31A1 and down-regulated ATP7B expression, which increased intracellular copper concentration, down-regulated FDX1, influenced the lipoylation of DLAT and DLD, and subsequently increased the expression of the stress protein HSP70 and the expression of PDHA1, inducing copper death in K562 cells. In addition, we investigated the toxicity of DEPS in vivo and demonstrated its low in vivo toxicity and adequate in vivo safety. In conclusion, our results suggest that DEPS may induce cuproptosis in cells, offering valuable insights for the future application of PS in leukemia treatment.

OBJECTIVE:To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.METHODS:MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.RESULTS:The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.CONCLUSION:DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.

Patrinia scabiosaefolia Fisch (PS), a perennial herb belonging to the genus Pinus in the family Pinnacle Sauce, has been previously known for its analgesic, anti-inflammatory, antibacterial, and antitumor properties. However, the specific mechanism behind its antileukemic effect remains unknown. This study focused on the cytotoxicity and potential modes of action of the dichloromethane extract from PS (DEPS) in acute myeloid leukemia (AML) cells. Our results demonstrated that DEPS reduced cell viability, arrested the cell cycle in the G2/M phase, disrupted the mitochondrial membrane potential, increased reactive oxygen species (ROS) production, and upregulated the expression of Bax/Bcl-2 and Cleaved caspase-3. However, the impact of DEPS on cell viability and the expression of apoptosis-associated proteins was reversed upon pretreatment with the caspase-3 inhibitor (Z-DEVD-FMK) in HL-60 cells, which demonstrated that DEPS could induce apoptosis through the mitochondria-associated apoptotic pathway. Interestingly, DEPS also influenced autophagy by upregulating the expression of LC3II/I, P62, and Beclin-1 proteins, and the autophagy inhibition chloroquine(CQ) could attenuate the apoptotic effects of DEPS in HL-60 cells. Furthermore, SMART 2.0 analysis predicted that the main components present in DEPS were likely terpenoids. In conclusion, DEPS possibly exerts antileukemic effects by downregulating the PI3K/AKT and ERK pathways, thereby promoting intracellular ROS production, activating the mitochondrial apoptotic pathway, and affecting autophagy, providing valuable insights for the potential future application of PS in the treatment of AML.

Around 11% of all known gene lesions causing human genetic diseases are nonsense mutations that introduce a premature stop codon (PTC) into the protein-coding gene sequence. Drug-induced PTC readthrough is a promising therapeutic strategy for treating hereditary diseases caused by nonsense mutations. To date, it has been found that more than 50 small-molecular compounds can promote PTC readthrough, known as translational readthrough-inducing drugs (TRIDs), and can be divided into two major categories: aminoglycosides and non-aminoglycosides. This review summarizes the pharmacodynamics and clinical application potential of the main TRIDs discovered so far, especially some newly discovered TRIDs in the past decade. The discovery of these TRIDs brings hope for treating nonsense mutations in various genetic diseases. Further research is still needed to deeply understand the mechanism of eukaryotic cell termination and drug-induced PTC readthrough so that patients can achieve the greatest benefit from the various TRID treatments.

针对病理生理学实验传统教学存在的问题,将成果导向教育理念引入临床医学专业病理生理学实验教学中,结合课程特点制定三维课程目标,构建混合式教学模式,采用多元考核方式,重视课堂互动与学生反馈,达成以学习结果 产出为导向的预期目标,持续改进教学质量,为实验教学改革提供思路.

Spectrin, as one of the major components of a plasma membrane-associated cytoskeleton, is a cytoskeletal protein composed of the modular structure of α and β subunits. The spectrin-based skeleton is essential for preserving the integrity and mechanical characteristics of the cell membrane. Moreover, spectrin regulates a variety of cell processes including cell apoptosis, cell adhesion, cell spreading, and cell cycle. Dysfunction of spectrins is implicated in various human diseases including hemolytic anemia, neurodegenerative diseases, ataxia, heart diseases, and cancers. Here, we briefly discuss spectrins function as well as the clinical manifestations and currently known molecular mechanisms of human diseases related to spectrins, highlighting that strategies for targeting regulation of spectrins function may provide new avenues for therapeutic intervention for these diseases.

Hereditary spherocytosis (HS) is the most frequently observed chronic non-immune hemolytic disorder caused by altered red cell membrane function. SPTB gene mutation is one of the most common causes of HS, but pathogenicity analyses and pathogenesis research on these mutations have not been widely conducted. In this study, a novel heterozygous mutation of the SPTB gene (c.1509_1518del; p.K503Nfs*67) was identified in a Chinese family with HS by whole-exome sequencing (WES) and was then confirmed by Sanger sequencing. Next, the pathogenicity and pathogenesis of this mutation were studied using peripheral blood. We found that this mutation disrupted the synthesis and localization of β-spectrin and weakened the interaction between β-spectrin and ankyrin, which may be caused by the nonsense-mediated mRNA degradation pathway. These changes lead to the transformation of discoid erythrocytes into spherocytes, resulting in hemolytic anemia. Therefore, we classified this novel mutation as a pathogenic mutation leading to loss-of-function of β-spectrin. It would be insightful to perform the same mutation test and to provide genetic counseling to other relatives of the proband. Our study increases the current understanding of the molecular mechanisms related to mutations in SPTB.

Background: Autophagy plays a vital role in cancer initiation, malignant progression, and resistance to treatment. However, autophagy-related genes (ARGs) have rarely been analyzed in gastric cancer (GC). The purpose of this study was to analyze ARGs in GC using bioinformatic analysis and to identify new biomarkers for predicting the overall survival (OS) of patients with GC. Methods: The gene expression profiles and clinical data of patients with GC were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets, and ARGs were obtained from two other datasets (the Human Autophagy Database and Molecular Signatures Database). Lasso, univariate, and multivariate Cox regression analyses were performed to identify the OS-related ARGs. Finally, a six-ARG model was identified as a prognostic indicator using the risk-score model, and survival and prognostic performance were analyzed based on the Kaplan-Meier test and ROC curve. Estimate calculations were used to assess the immune status of this model, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were employed for investigating the functions and terms associated with the model-related genes in GC. Results: The six ARGs, DYNLL1, PGK2, HPR, PLOD2, PHYHIP, and CXCR4, were identified using Lasso and Cox regression analyses. Survival analysis revealed that the OS of GC patients in the high-risk group was significantly lower than that of the low-risk group (p < 0.05). The ROC curves revealed that the risk score model exhibited better prognostic performance with respect to OS. Multivariate Cox regression analysis indicated that the model was an independent predictor of OS and was not affected by most of the clinical traits (p < 0.05). The model-related genes were associated with immune suppression and several biological process terms, such as extracellular structure organization and matrix organization. Moreover, the genes were associated with the P13K-Akt signaling pathway, focal adhesion, and MAPK signaling pathway. Conclusions: This study presents potential prognostic biomarkers for GC patients that would aid in determining the best patient-specific course of treatment.

根据病理生理学教学现状及存在的问题,创建并应用智慧课堂融入课程思政教学模式.通过对该教学模式创新内涵及实施过程的介绍,深层次剖析其在病理生理学教学中的应用价值及实际意义,为今后病理生理学教学改革提供参考.

Phelan-McDermid syndrome (PMS)(OMIM#606232) is a rare genetic disorder caused by a deletion of the distal long arm of chromosome 22q13 involving a variety of clinical features with considerably heterogeneous degrees of severity. This syndrome is characterized by global developmental delay, intellectual disability, hypotonia, absent or severely delayed speech, minor dysmorphic features and autism spectrum disorder. PMS is easy to be misdiagnosed due to the lack of specific clinical manifestations. SHANK3 has been identified as the critical candidate gene for the neurological features of this syndrome. However, some studies have shown that other genes located in the 22q13 region may have a role in the formation of symptoms in individuals with PMS. This article provides a review for recent progress made in research on PMS including etiology, clinical manifestation, diagnosis, and treatment, with a particular emphasis on clinical diagnosis and treatment.

新型冠状病毒肺炎疫情的爆发,体现出基础研究与临床应用双向转化的重要性,即转化医学,依托于多学科相互渗透.病理生理学是连接基础与临床的桥梁学科,与转化医学极具共性,借助于医学各学科相互交叉.因此,转化医学理念指导病理生理学教学,采用针对性的教学策略,培养学生科研思维及临床判断力,形成基于学科交叉的转化医学教学模式.文章对本教学模式的理论依据、教学策略及培养目标予以梳理,并浅评教学效果,提出改进建议,为各基础医学学科教学提供新思路,推动医学教育创新发展.

Background: Exosomes play an important role in transferring information among different cell types, as they transport materials from the cell membrane to the cytoplasm. They are involved not only in normal physiological functions, but also in the occurrence and development of a variety of diseases. Cancer is a major health problem affecting humans. Currently, exosomes are considered novel stars in tumor therapy. Objective: To present a review focusing on the role of exosomes in tumorigenesis and development and the possibility of treating tumors with exosome-targeted therapies or using exosomes as carriers. Methods: We reviewed literature related to the biological origin and function of exosomes and exosome-tumor relationship. Results: Exosomes are closely related to tumor immunity, angiogenesis, pre-metastasis microenvironment, chemoresistance, energy metabolism, etc. Tumor therapy involving the targeting of exosomes involves block the generation, secretion, uptake of exosomes, and elimination of circulating exosomes, and develop antitumor vaccines. Exosome as delivery vehicles can be loaded with chemotherapeutic drugs, therapeutic genes, and other therapeutic drugs to target cells. Prospects and challenges of exosome-based tumor therapy are also discussed. Conclusion: Exosomes are involved in multiple processes during tumor development and should be further studied as novel targets for cancer therapy.

Objective: To investigate the relationship between cyclooxygenase-2 (COX-2) and nuclear factor-related factor-2 (NF-2) in liver tissue and the degree of inflammation in (NAFLD) rats with nonalcoholic fatty liver disease. Methods: Fifty adult rats were randomly divided into experimental group (n=30), control group (n=10) and Aspirin intervention group (n=10). The experimental group was divided into a five- week experimental group, 10-week experimental group and a 15-week experimental group according to the prolongation of feeding time. The rats in the experimental group were fed with high fat emulsion, while those in the control group were given normal saline and free access to common feed and drinking water. The Aspirin intervention group was given Aspirin at 10Mg/100g via intragastrical feeding with high fat emulsion. The body weight, liver size, shape, texture, liver index and liver tissue of rats in each group were measured. The degree of liver inflammation was observed by HE staining. The stage of inflammation was judged according to the diagnostic criteria of histology. At the same time, the expression of cyclooxygenase-2 (COX-2) and nuclear factor-related factor-2 (Nrf-2) were detected by monoclonal antibodies, and the correlation with the inflammatory process was analysed. Results: The physical weight of each group from large to small was 15 weeks in the experimental group, Aspirin intervention group, 10 weeks in the experimental group and ,five weeks in the experimental group. The difference was statistically significant (P<0.05), and the liver wet weight and LI from large to small were 15 weeks in the experimental group, Aspirin intervention group, 10 weeks in the experimental group, five weeks in the experimental group, the control group and the difference was again statistically significant (P<0.05). In the control group, COX-2 and Nrf-2 were weakly positive in the cytoplasm and capsule of very few hepatocytes. COX-2 and Nrf-2 appeared in the cytoplasm of the damaged hepatocytes in the experimental group at five weeks, the positive cells in the experimental group increased again, the positive cells in the damaged cells were moderately positive in the 10 groups and the positive cells in the Aspirin intervention group were diffusible. They were higher than those in the experimental group at 10 weeks. At the 15th week, the expression of COX-2 and Nrf-2 in the experimental group was the strongest, followed by the Aspirin intervention group, followed by the experimental group at the 10th week, the experimental group at the fifth week, and the control group with the weakest expression of COX-2 and Nrf-2. The difference was statistically significant (P<0.05). In the control group, the hepatic lobule structure of the normal liver tissue of the male rats was normal, and the morphology of the hepatocytes was normal; the liver cells of the experimental group were mostly fatty lesions, which were the simple fatty lesions and the liver cells of the experimental group. The experimental group and the Aspirin intervention group were more than four points, which was in accordance with the judgment of NASH. The liver cells of the experimental group and the Aspirin intervention group were more than four points at the 10th week, the 15th week of the experimental group and the control group. At 15 weeks, the hepatocytes of the experimental group were diffusely adipose; the cells were enlarged and varied in size. They were also accompanied by balloon transformation of hepatocytes and infiltration of inflammatory cells in the lobule. Conclusion: The expression of COX-2 and Nrf-2 in the liver tissue of rats with alcoholic fatty liver disease reflects the inflammatory process of liver tissue to a certain extent, and it is of great clinical significance to evaluate the inflammatory process of liver tissue.

Background: Exosomes play an important role in transferring information among different cell types, as they transport materials from the cell membrane to the cytoplasm. They are involved not only in normal physiological functions, but also in the occurrence and development of a variety of diseases. Cancer is a major health problem affecting humans. Currently, exosomes are considered novel stars in tumor therapy. Objective: To present a review focusing on the role of exosomes in tumorigenesis and development and the possibility of treating tumors with exosome-targeted therapies or using exosomes as carriers. Methods: We reviewed literature related to the biological origin and function of exosomes and exosome-tumor relationship. Results: Exosomes are closely related to tumor immunity, angiogenesis, pre-metastasis microenvironment, chemoresistance, energy metabolism, etc. Tumor therapy involving the targeting of exosomes involves block the generation, secretion, uptake of exosomes, and elimination of circulating exosomes, and develop antitumor vaccines. Exosome as delivery vehicles can be loaded with chemotherapeutic drugs, therapeutic genes, and other therapeutic drugs to target cells. Prospects and challenges of exosome-based tumor therapy are also discussed. Conclusion: Exosomes are involved in multiple processes during tumor development and should be further studied as novel targets for cancer therapy.