The traditional theory of "the spleen governs the dispersion of essence" refers to the spleen's pivotal role in distributing refined nutrients throughout the body. In traditional Chinese medicine （TCM）， lipids are categorized under "gaozhi （膏脂）"， and their transportation and metabolism via apolipoproteins are believed to be closely related to the spleen's dispersing function. The liver， which synthesizes apolipoproteins， is functionally linked to the spleen system in TCM. Impaired dispersion of essence by the spleen and disrupted transportation of gaozhi constitute the pathological mechanism of dyslipidemia due to spleen deficiency. Strengthening the spleen and resolving dampness is the core therapeutic principle. From the perspective of modern medicine， this may involve promoting hepatic functional recovery related to lipid metabolism， thereby enhancing lipid processing and reducing the levels of abnormally accumulated lipids in the bloodstream.

ObjectiveTo construct and validate a clinical prediction model for Qi deficiency and blood stasis syndrome in chronic heart failure （CHF），aiming to assist clinical diagnosis and provide tools and methods for individualized treatment of CHF. MethodsThe clinical data of patients with chronic heart failure treated at Dongzhimen Hospital of Beijing University of Chinese Medicine from January 2022 to January 2024 were retrospectively collected. The patients were randomly divided into a training group and a validation group with a ratio of 7∶3. First， the least absolute shrinkage and selection operator （LASSO） regression analysis was used to preliminarily screen the predictive factors affecting the diagnosis of Qi deficiency and blood stasis syndrome in CHF. Subsequently， the Logistic regression method was applied to conduct a more in-depth and detailed analysis of these factors. Variables with P<0.05 in the results of the multi-factor Logistic regression were carefully selected and included. Based on the regression coefficients obtained from this analysis， a model was constructed， and a nomogram was accurately drawn. Using R software，the receiver operating characteristic （ROC） curve，calibration curve，and decision curve analysis （DCA） were precisely drawn. These analyses were used to comprehensively evaluate the model from three crucial aspects： discrimination，calibration，and clinical applicability. Additionally， the accuracy，specificity，sensitivity，positive predictive value，and negative predictive value of the model were meticulously calculated to conduct a more all-round and comprehensive assessment. ResultsIn total， 168 cases were successfully obtained in the training group， and 71 cases were included in the validation group. After a thorough comparison， it was found that there were no statistically significant differences in the baseline data between the two groups. After being rigorously screened by the LASSO-multivariate logistic regression method， dark red tongue，smoking history，cardiac troponin I，and N-terminal pro-B-type natriuretic peptide （NT-ProBNP） were identified as the influencing factors for diagnosing patients with the Qi deficiency and blood stasis syndrome in CHF. The constructed model demonstrated an area under the curve （AUC） of 0.812 in the training group and 0.719 in the validation group. The calibration curve showed that the predicted curve of the model was close to the actual observed curve. DCA indicated that the model could provide substantial clinical benefits for patients at the decision thresholds ranging from 0.2 to 0.9. ConclusionThe clinical prediction model for Qi deficiency and blood stasis syndrome in chronic heart failure constructed in this study shows good performance. It has certain application value in clinical practice， which may contribute to the improvement of the diagnosis and treatment of CHF patients with this syndrome.

ObjectiveTo observe the effectiveness and safety of Jiuzihuichun decoction combined with spleen- strengthening moxibustion in patients with asthenospermia infertility with spleen-kidney deficiency pattern. MethodsA total of 82 patients with asthenospermia of spleen-kidney deficiency pattern in Shanghai Seventh People's Hospital were included. The patients were randomly divided into an observation group and a control group， with 41 patients in each group. The control group received oral administration of WuziYanzong pills combined with spleen-strengthening moxibustion. The dosage of Wuzi Yanzong pills was 1 bag each time， and it was taken twice a day. The spleen-strengthening moxibustion was carried out once a week. The observation group， on the other hand， took Jiuzi Huichun decoction orally combined with spleen-strengthening moxibustion. The Jiuzi Huichun Decoction was taken 200 mL each time， twice a day， with one dose in the morning and one in the evening. The spleen-strengthening moxibustion for the observation group was also performed once a week. The treatment course for both groups was 12 weeks， and they were followed up for an additional 12 weeks. During the treatment process，12 cases were either lost to follow-up or excluded. Eventually， 70 cases were available for evaluation，with 35 cases in the control group and 35 cases in the observation group. The pregnancy status of the patients' spouse within 6 months was recorded. The traditional Chinese medicine （TCM） syndrome scores of spleen-kidney deficiency pattern before and after treatment were evaluated. The semen volume，semen routine parameters，normal sperm morphology，sperm DNA fragmentation index，seminal plasma fructose，seminal plasma acid phosphatase， and seminal plasma α-glucosidase levels of the two groups were detected before and after treatment. In addition， the safety indicators related to liver and kidney functions of the two groups were detected before and after treatment. ResultsDuring the 6-month observation period， when compared with the situation before treatment in their respective groups，the semen volume of the observation group and the control group increased. In contrast， the sperm concentration，sperm motility，proportion of a+b-grade sperm，normal sperm morphology，seminal plasma fructose，seminal plasma acid phosphatase，seminal plasma α-Glucosidase，the proportion of a-grade sperm，linear sperm motility，linear sperm concentration， and linear sperm count all increased significantly（P<0.05）. At the same time， the sperm DNA fragmentation index and the TCM syndrome scores of the spleen-kidney deficiency pattern decreased significantly（P<0.05）. When the observation group was compared with the control group after treatment， the clinical efficacy of the observation group was better（Z=-2.276，P<0.05）. The pregnancy rate of the observation group's spouses was 14.3%，which was higher than the 2.9% of the control group. The sperm motility， the proportion of a+b-grade sperm，seminal plasma fructose，the proportion of a-grade sperm，normal sperm morphology，α-glucosidase， and linear sperm motility in the observation group were higher than those in the control group （P<0.05）. Moreover， the sperm DNA fragmentation index and the TCM syndrome scores of spleen-kidney deficiency pattern in the observation group were lower than those in the control group （P<0.01）. No serious adverse reactions occurred in the two groups，and no abnormalities were found in the safety indicators after treatment. ConclusionJiuzi Huichun decoction combined with spleen-strengthening moxibustion can enhance sperm viability and sperm concentration. It can also improve the TCM-related symptoms of asthenospermia of spleen-kidney deficiency pattern and sperm morphology. Additionally， it can reduce the sperm DNA fragmentation index and regulate the level of seminal plasma bioenzyme in patients with male asthenospermia infertility of spleen-kidney deficiency pattern. Therefore， it is worthy of further promotion and application in clinical practice.

BACKGROUND: T-cell lymphoblastic lymphoma/acute lymphoblastic leukemia (T-LBL/ALL) is an aggressive form of hematological malignancy associated with poor prognosis in adult patients. Histone deacetylases (HDACs) are aberrantly expressed in T-LBL/ALL and are considered potential therapeutic targets. Here, we investigated the antitumor effect of a novel HDAC inhibitor, chidamide, on T-LBL/ALL. METHODS: HDAC1, HDAC2 and HDAC3 levels in T-LBL/ALL cell lines and patient samples were compared with those in normal controls. Flow cytometry, transmission electron microscopy, and lactate dehydrogenase release assays were conducted in Jurkat and MOLT-4 cells to assess apoptosis and pyroptosis. A specific forkhead box O1 (FOXO1) inhibitor was used to rescue pyroptosis and upregulated gasdermin E (GSDME) expression caused by chidamide treatment. The role of the FOXO1 transcription factor was evaluated by dual-luciferase reporter and chromatin immunoprecipitation assays. The efficacy of chidamide in vivo was evaluated in a xenograft mouse. RESULTS: The expression of HDAC1, HDAC2 and HDAC3 was significantly upregulated in T-LBL/ALL. Cell viability was obviously inhibited after chidamide treatment. Pyroptosis, characterized by cell swelling, pore formation on the plasma membrane and lactate dehydrogenase leakage, was identified as a new mechanism of chidamide treatment. Chidamide triggered pyroptosis through caspase 3 activation and GSDME transcriptional upregulation. Chromatin immunoprecipitation assays confirmed that chidamide led to the increased transcription of GSDME through a more relaxed chromatin structure at the promoter and the upregulation of FOXO1 expression. Moreover, we identified the therapeutic effect of chidamide in vivo . CONCLUSIONS This study suggested that chidamide exerts an antitumor effect on T-LBL/ALL and promotes a more inflammatory form of cell death via the FOXO1/GSDME axis, which provides a novel choice of targeted therapy for patients with T-LBL/ALL.
Humans ; Pyroptosis/drug effects* ; Forkhead Box Protein O1/genetics* ; Aminopyridines/pharmacology* ; Animals ; Mice ; Benzamides/pharmacology* ; Cell Line, Tumor ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Phosphate-Binding Proteins/metabolism* ; Histone Deacetylase Inhibitors/pharmacology* ; Jurkat Cells ; Histone Deacetylases/metabolism* ; Apoptosis/drug effects* ; Gasdermins

Objective To explore the function of olfactomedin domain family protein 3(OLFM3)in neuroblastoma(NB).Methods The relationship between the expression of OLFM3 mRNA and v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog(MYCN)amplification status and the prognosis of patients in NB clin-ical samples were clarified by using R2 Genomics Analysis and Visualization Platform.Depmap database was used to examine the expression level of OLFM3 in different tumors cell lines and to identify the correlation between OLFM3 expression and MYCN amplification status in various NB cell lines.RT-qPCR and Western blot were used to detect the knockdown level of OLFM3.Cell proliferation was monitored using crystal violet staining and real?time cellular analysis.The colony formation ability of NB cells was assessed using colony?forming unit assay.Results Analysis of R2 database revealed higher level of OLFM3 expression in MYCN?amplified NB clinical samples(P<0.001).Patients with high OLFM3 expression showed a significantly lower overall survival probability compared to those with low OLFM3 expression(P<0.05).Analysis with Depmap database revealed that the expres?sion level of OLFM3 was higher in NB than that in other kind of tumor.The expression level of OLFM3 was signifi?cantly higher in the MYCN?amplified cell lines than in the MYCN?non?amplified cell lines(P<0.01).In MYCN?am?plified NB cells,knockdown of OLFM3 inhibited cells proliferation(P<0.001)and colony formation(P<0.001),but there was no noticeable changes observed in MYCN?non?amplified cells.Conclusions OLFM3 specifically pro?motes the proliferation of MYCN?amplified NB cells,but has a less effect on MYCN?non?amplified cells,indicating it is a potential biomarker for high?risk MYCN?amplified NB.

OBJECTIVES: To investigate the role of PTEN-induced putative kinase 1 (PINK1) in regulating the viability, migration, and apoptosis of colorectal cancer (CRC) cells, and to explore its potential epigenetic mechanisms. METHODS: PINK1 was overex-pressed or knocked down in HCT116 and DLD1 CRC cell lines using lentiviral vectors, with efficiency verified by qRT-PCR and Western blotting. Cell proliferation, colony formation, migration, and apoptosis were assessed using CCK-8, colony formation, wound healing, Transwell, and Hoechst 33258 staining assays, respectively. Protein levels of apoptosis-related and histone modification-related markers were analyzed by Western blotting. Genome-wide chromatin accessibility was profiled using ATAC-seq. RESULTS: PINK1 expression was significantly downregulated at both mRNA and protein levels in CRC tissues compared to normal tissues. PINK1 overexpression inhibited cell prolifera-tion, colony formation, and migration in HCT116 and DLD1 cells (all P<0.05), whereas PINK1 knockdown promoted these malignant phenotypes (all P<0.05). PINK1 overex-pression induced apoptosis, associated with decreased levels of anti-apoptotic proteins (Mcl-1, Bcl-2, Bcl-xl) and increased pro-apoptotic Bax (all P<0.05), without altering p53 expression. Mechanistically, PINK1 overexpression reduced the expression of histone H3 lysine 9 trimethylation (H3K9me3) and histone H3 lysine 27 trimethylation (H3K27me3), and increased the expression of histone H3 lysine 9 acetylation (H3K9ac) and histone H3 lysine 27 acetylation (H3K27ac). It also downregulated key histone-modifying enzymes, including EZH2, EZH1, SUZ12, and histone deacetylase 3 (HDAC3) (all P<0.01). ATAC-seq revealed that PINK1 overexpression increased chromatin accessibility, particularly around transcription start sites. CONCLUSIONS PINK1 acts as a tumor suppressor in colorectal cancer by inhibiting proliferation and migration, promoting apoptosis, and remodeling the epigenetic landscape through altering histone modifications and enhancing chromatin accessibility.

Insufficient alveolar bone thickness increases the risk of periodontal dehiscence and fenestration, especially in orthodontic tooth movement. Abaloparatide (ABL), a synthetic analog of human PTHrP (1-34) and a clinical medication for treating osteoporosis, has recently demonstrated its potential in enhancing craniofacial bone formation. Herein, we show that intraoral submucosal injection of ABL, when combined with mechanical force, promotes in situ alveolar bone thickening. The newly formed bone is primarily located outside the original compact bone, implying its origin from the periosteum. RNA sequencing of the alveolar bone tissue revealed that the focal adhesion (FA) pathway potentially mediates this bioprocess. Local injection of ABL alone enhances cell proliferation, collagen synthesis, and phosphorylation of focal adhesion kinase (FAK) in the alveolar periosteum; when ABL is combined with mechanical force, the FAK expression is upregulated, in line with the accomplishment of the ossification. In vitro, ABL enhances proliferation, migration, and FAK phosphorylation in periosteal stem cells. Furthermore, the pro-osteogenic effects of ABL on alveolar bone are entirely blocked when FAK activity is inhibited by a specific inhibitor. In summary, abaloparatide combined with mechanical force promotes alveolar bone formation via FAK-mediated periosteal osteogenesis. Thus, we have introduced a promising therapeutic approach for drug-induced in situ alveolar bone augmentation, which may prevent or repair the detrimental periodontal dehiscence, holding significant potential in dentistry.
Osteogenesis/drug effects* ; Periosteum/cytology* ; Parathyroid Hormone-Related Protein/administration & dosage* ; Animals ; Focal Adhesion Protein-Tyrosine Kinases/metabolism* ; Alveolar Process/drug effects* ; Cell Proliferation/drug effects* ; Phosphorylation ; Rats ; Male ; Humans ; Focal Adhesion Kinase 1/metabolism* ; Cell Movement/drug effects*

Objective To investigate the mechanism of argon-helium cryotherapy in regulating the tyrosine-protein kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)path-way,thereby inhibiting the inflammatory response in rats with atherosclerotic occlusive disease(ASO)of the lower extremities.Methods Forty rats were randomly divided into sham group,lower extremity ASO group,Ar-He Cryo group and Ar-He Cryo+p-JAK group,with 10 rats in each group.The average blood flow velocity of the lower extremity artery,lipid profile and inflammation levels were measured in each group.Hematoxylin-eosin(HE)staining was used to examine histopathologi-cal changes in arterial vessel tissues.Western blotting was performed to detect the expression levels of JAK2,p-JAK2,STAT3,p-STAT3 and suppressor of cytokine signaling 3(SOCS3)proteins in arterial vessel tissues.Results The average blood flow velocity of the lower extremity artery in the lower extremity ASO group was significantly lower than that in the sham group(P＜0.05).The average blood flow velocity in the Ar-He Cryo group was significantly higher than that in the ASO group(P＜0.05).The average blood flow velocity in the Ar-He Cryo+p-JAK group was significantly lower than that in the Ar-He Cryo group(P＜0.05).Serum total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C)levels,tumor necrosis factor-alpha(TNF-α),interleukin-1 β(IL-1β),interleukin-6(IL-6)levels and endothelin-1(ET-1)content in the lower extremity ASO group were significantly higher than those in the sham group,while serum high-densi-ty lipoprotein cholesterol(HDL-C)levels and nitric oxide(NO)content were significantly lower than those in the sham group(P＜0.05).In the Ar-He Cryo group,serum TC,TG,LDL-C levels as well as TNF-α,IL-1β,IL-6 levels and ET-1 content were significantly lower than those in the lower extremity ASO group,while serum HDL-C levels and NO content were significantly higher(P＜0.05).In the Ar-He Cryo+p-JAK group,serum TC,TG,LDL-C levels as well as TNF-α,IL-1 β,IL-6 levels and ET-1 content were significantly higher than those in the Ar-He Cryo group,while HDL-C levels and NO content were significantly lower(P＜0.05).Compared with the sham group,the lower extremity venous vessels in the lower extremity ASO group showed significant steno-sis;compared with the lower extremity ASO group,the degree of stenosis in the lower extremity ve-nous vessels was significantly reduced in the Ar-He Cryo group;the degree of stenosis in the lower extremity venous vessels in the Ar-He Cryo+p-JAK group was similar to that in the lower extremity ASO group and significantly more severe than that in the Ar-He Cryo group.The ratios of p-JAK2/JAK2 and p-STAT3/STAT3 in arterial vessel tissues of the lower extremity ASO group were signifi-cantly higher than those in the sham group,and SOCS3 protein expression was significantly lower than that in the sham group(P＜0.05).In the Ar-He Cryo group,p-JAK2/JAK2 and p-STAT3/STAT3 in arterial vessel tissues were significantly lower than those in the lower extremity ASO group,and SOCS3 protein expression was significantly higher than that in the lower extremity ASO group(P＜0.05).In the Ar-He Cryo+p-JAK group,p-JAK2/JAK2 and p-STAT3/STAT3 in arte-rial vessel tissues were significantly higher than those in the Ar-He Cryo group,and SOCS3 protein expression was significantly lower than that in the Ar-He Cryo group(P＜0.05).Conclusion Ar-gon-helium cryotherapy can reduce lipid deposition in the arteries of rats with lower extremity ASO,inhibit inflammatory responses,and its mechanism may be related to the inhibition of activation of JAK2/STAT3 signaling pathway.

Objective To explore the effect of open reading frame 66(C12ORF66)located at chromosome 12 on the viability of MYCN amplified NB cell lines.Methods DDatasets GSE16476 and GSE49710 in R2 database were analyzed for expression level of C12ORF66 in MYCN amplified and MYCN non-amplified NB cells and its potential correlation with the prognosis of pediatric patients.C12ORF66 mRNA expression level in normal tissue immortalized cell lines,MYCN amplified and MYCN non-amplified cell lines were detected by RT-qRCR.Transient or stable knockdown of C12ORF66 cell lines were constructed to compare the difference in real time cellular analysis(RTCA),colony formation,Ki67 positive cells between the control group and the C12ORF66 knockdown group.Results By analyzing R2 datasets,C12ORF66 level in MYCN amplified samples was significantly higher than that in MYCN non-amplified samples,and the expression of C12ORF66 was negatively correlated with the prognosis of pediatric patients(P＜0.05).C12ORF66 highly expressed in MYCN-amplified BE(2)-C and SK-N-BE(2)cell lines than in MYCN non-amplified CHLA-255 and SH-SY5Y cell lines(P＜0.001).Transient or stable knockdown of C12ORF66 resulted in significant slow down of proliferation of MYCN amplified NB cells(P＜0.001),the colony formation ability was significantly reduced(P＜0.001),and the proportion of Ki67 positive cells was significantly decreased(P＜0.05).Conclusions C12ORF66 was highly expressed in MYCN amplified clinical NB samples and cell lines which is believed to be correlated with poor prognosis of pediatric patients.C12ORF66 knockdown signifi-cantly inhibits cell viability of NB cells.

Objective To investigate the regulatory effects of wedelobata on apoptosis and secretion of inflammatory factors in the alveolar epithelial cells infected by Streptococcus pn e um on i a e.Methods Alveolar epithelial cells A549 were divided into infection group(1×108/CFU/mL cultured cells of Streptococcus pneumoniae),control group(no treatment),infection+wedelolactone low-dose group,middle-dose group and high-dose group(pretreated with wedelolactone at 10,20 and 40 μmol/L and then cultured cells of Streptococcus pneumoniae at 1×108/CFU/mL).Alveolar epithelial cells A549 were divided into infection group(1×108/CFU/mL cultured cells of Streptococcus pneumoniae),control group(no treatment),infection+wedelolactone low-dose group,middle-dose group and high-dose group(pretreated with wedelolactone at 10,20 and 40 μmol/L and then cultured cells of Streptococcus pneumoniae at 1×108/CFU/mL).Results Compared with control group,the apoptosis rate,the relative expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA were higher in infection group,infection + wedelolide low dose group,medium dose group and high dose group,while the expression level of Bcl-2 protein was lower(P＜0.05).Compared with the infected group,the apoptosis rate,the relative expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA and the expression levels of Bcl-2 protein were lower in the infected + wedelolide low dose,medium dose and high dose groups.Moreover,the apop-tosis rate,the expression levels of Bax,Caspase-3 protein,IL-6,IL-1β and TNF-α mRNA were the highest in the infected + wedelactone high-dose group,and the expression levels of Bcl-2 protein were the lowest(P＜0.05).Conclusion The apoptosis rate of alveolar epithelial cells infected by Streptococcus pneumoniae decreased and the secretion of inflammatory factors decreased after the intervention of wedelia lactone.