The cardioprotective effects of histone deacetylase (HDAC) inhibitors (HDIs) are at odds with the deleterious effects of HDAC depletion. Here, we use HDAC3 as a prototype HDAC to address this contradiction. We show that adult-onset cardiac-specific depletion of HDAC3 in mice causes cardiac hypertrophy and contractile dysfunction on a high-fat diet (HFD), excluding developmental disruption as a major reason for the contradiction. Genetically abolishing HDAC3 enzymatic activity without affecting its protein level does not cause cardiac dysfunction on HFD. HDAC3 depletion causes robust downregulation of lipid oxidation/bioenergetic genes and upregulation of antioxidant/anti-apoptotic genes. In contrast, HDAC3 enzyme activity abolishment causes much milder changes in far fewer genes. The abnormal gene expression is cardiomyocyte-autonomous and can be rescued by an enzyme-dead HDAC3 mutant but not by an HDAC3 mutant (Δ33-70) that lacks interaction with the nuclear-envelope protein lamina-associated polypeptide 2β (LAP2β). Tethering LAP2β to the HDAC3 Δ33-70 mutant restored its ability to rescue gene expression. Finally, HDAC3 depletion, not loss of HDAC3 enzymatic activity, exacerbates cardiac contractile functions upon aortic constriction. These results suggest that the cardiac function of HDAC3 in adults is not attributable to its enzyme activity, which has implications for understanding the cardioprotective effects of HDIs.

OBJECTIVES: To explore the therapeutic mechanism of Guizhi Fuling (GZFL) Pellets against cervical cancer. METHODS: Publicly available databases were used to identify the targets of GZFL Pellets and cervical cancer to construct the protein-protein interaction (PPI) network, followed by GO biological process and KEGG pathway enrichment analysis of the hub genes. The "Traditional Chinese Medicine-Active Ingredients-Targets-Pathways" network for GZFL Pellets in cervical cancer treatment was generated using Cytoscape v10.0.0, and molecular docking of the drug and potential targets was performed to predict the specific targets of active components in Guizhi Fuling Pellets. The inhibitory effects of hederagenin, an active ingredient in GZFL Pellets, was tested in cultured cervical cancer cells and in nude mice bearing cervical cancer xenografts. RESULTS: GZFL Pellets contain 338 active components targeting 247 action sites. A total of 10127 cervical cancer-related targets were obtained, and among them 195 were identified as potential therapeutic targets of GZFL Pellets for cervical cancer treatment, including the key targets of GABRA1, PTK2, JAK2, HTR3A, GSR, and IL-17. Molecular docking study showed low binding energies of the active components such as hederagenin, campesterol, and stigmasterol for protein-molecule interaction. GO enrichment analysis suggested that GZFL Pellets inhibited cervical cancer primarily by regulating responses to steroid hormones, oxidative stress, and lipopolysaccharides. Among the active components of GZFL Pellets, hederagenin was found to inhibit cervical cancer cells in vitro and significantly reduced STAT3 phosphorylation level in the cancer cells. In nude mice bearing cervical cancer xenografts, hederagenin effectively inhibited tumor growth rate without causing obvious adverse effects. CONCLUSIONS GZFL Pellets inhibit cervical cancer cell growth through its multiple active components that target different pathways. Among these components, hederagenin inhibits tumor cell growth possibly by directly binding to JAK2 protein to inhibit STAT3 phosphorylation.
Female ; Animals ; Uterine Cervical Neoplasms/pathology* ; Mice, Nude ; Humans ; Mice ; Oleanolic Acid/therapeutic use* ; Drugs, Chinese Herbal/therapeutic use* ; Molecular Docking Simulation ; Xenograft Model Antitumor Assays ; Cell Line, Tumor ; STAT3 Transcription Factor/metabolism* ; Protein Interaction Maps ; Janus Kinase 2/metabolism*

Objective:To evaluate the treatment strategies and outcomes for contracted nasal deformity.Methods:A retrospective review was conducted of the patients with contracted noses who underwent surgery at the Department of Plastic and Aesthetic Surgery, the Second Xiangya Hospital of Central South University, between January 2021 and January 2024. Based on the pathologic-anatomic features and severity of the deformity, patients were categorized as having mild, moderate or severe contraction. Mild cases received extensive subcutaneous dissection and framework reconstruction with various grafts. Moderate cases underwent preoperative nasal-skin distraction and intra-operative reconstruction with lower lateral or septal cartilage. Severe cases, in addition to reconstruction of the lower lateral cartilage and nasal septum, underwent individualized repair, including mucosal or cutaneous defect resurfacing. Postoperative follow-up assessed nasal appearance, complications and recurrence of contraction. Nasal aesthetics were quantified with visual analogue scale (VAS, 0-10 points; higher scores = less deformity), and patient satisfaction with the rhinoplasty outcome evaluation (ROE) questionnaire (0 = very dissatisfied, 100 = very satisfied) before surgery and at final follow-up. Paired t-test was used for VAS and ROE comparisons; categorical variables were analyzed with the χ2 test. A P-value < 0.05 denoted statistical significance. Results:A total of 96 patients were included, comprising 7 males and 89 females, aged (27.4 ± 8.0) years (19-58 years). There were 64 cases of mild, 19 cases of moderate, and 13 cases of severe contracted nose deformities. Postoperative follow-up(14.0 ± 4.6) months (6-19 months). No cases of abnormal nasal appearance, infection, necrosis, scar hyperplasia, or recurrence of contracted nose were observed during follow-up. Significant improvements in nasal aesthetic outcomes were observed. The VAS scores for mild, moderate, and severe contracted noses increased significantly from preoperative values of 4.7 ± 1.0, 3.0 ± 1.2, and 2.2± 1.1 to postoperative values of 8.6 ± 0.7, 8.9 ± 0.7, and 8.2 ± 0.9, respectively (all P < 0.01). Similarly, the ROE scores improved significantly from 59.1 ± 10.0, 34.2 ± 12.1, and 28.5± 6.3 preoperatively to 90.2 ± 9.5, 91.5 ± 7.5, and 93.3 ± 5.8 postoperatively (all P < 0.01). Conclusion:Selecting appropriate surgical methods based on the pathological and anatomical characteristics and severity of contracted nose deformities can achieve favorable outcomes. Postoperative nasal aesthetic appearance is significantly improved, and patient satisfaction is high.

Objective:To investigate the protective effect of myrislignan（MRL）on concanavalin A（Con A）-induced autoimmune hepatitis（AIH）.Methods:C57BL/6J mice were divided into the following groups using a random number table，with five mice in each group：control group，MRL group，model group（Con A group），and MRL pretreatment group（MRL+Con A group）. MRL was injected intraperitoneally at a dose of 30 μg/g；3 h after pretreatment，Con A（18 μg/g）was administrated by intravenous injection；mouse livers and serum samples were collected 12 h after injection for measuring serum transaminase levels and liver cell apoptosis. The mRNA and protein expression levels of IL-6，IL-12，and TNF-α were measured using qRT-PCR and ELISA. Flow cytometry was performed to detect the proportion and activation status of macrophages in liver tissues. Bone marrow-derived macrophages（BMDMs）were isolated and induced in vitro to analyze the regulatory effect of MRL on macrophages. One-way analysis of variance was used to compare the differences in various indicators among groups. Results:Compared with the Con A group，MRL（30 μg/g）pretreatment significantly reduced alanine aminotransferase（ P<0.05）and aspartate transaminase（ P<0.01）levels，attenuated liver oxidative stress（increased superoxide dismutase activity，while decreased levels of malondialdehyde and myeloperoxidase；all P<0.05），and suppressed hepatocyte apoptosis（ P<0.01）. Both in vivo and in vitro experiments confirmed that MRL（30 μg/g）could reduce the proportion of M1 macrophages（ in vivo： P<0.05； in vitro：all P<0.001）and inhibit macrophage activation（ in vivo： P<0.01； in vitro：all P<0.05）. Conclusion:MRL effectively prevents Con A-induced autoimmune hepatitis by inhibiting liver cell apoptosis，attenuating liver oxidative stress，suppressing M1 macrophage polarization，and reducing inflammatory cytokine expression.

Objective:To evaluate the role of stimulator of interferon genes (STING) in postoperative cognitive dysfunction (POCD) and the relationship with pyroptosis in hippocampal cells in aged mice.Methods:Forty-eight SPF healthy male C57BL/6 mice, aged 18 months, weighing 23-28 g, were assigned to 4 groups ( n=12 each) using a random number table method: control group (C group), POCD group (P group), STING inhibitor C-176 group (PC group), and C-176 solvent group (PV group). The mice underwent Morris water maze training for 4 days prior to model establishment. Mice in P, PC and PV groups underwent tibial fracture and intramedullary pin fixation under sevoflurane anesthesia to establish the POCD model, while mice in C group received no treatment. The STING inhibitor C-176 (750 nmol/200 μl) and an equal volume of C-176 solvent were intraperitoneally injected at 30 min before establishment of the model in PC and PV groups, respectively. The open field test was performed on the 5th day after model preparation, the novel object recognition test was conducted on the 6th day, and the Morris water maze test was performed on the 7th day. Mice were sacrificed under anesthesia to collect the hippocampus for determination of the expression of STING, phosphorylated STING (p-STING), nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), cleaved caspase-1, and gasdermin-D (GSDMD)-NT by Western blot. Results:There were no statistically significant differences in the parameters of the training phase of the Morris water maze test and the open field test among the four groups ( P>0.05). Compared with C group, the recognition index in the novel object recognition test was significantly decreased, the number of crossing the original platform was reduced and the duration spent in the target quadrant was shortened in the Morris water maze test, and the expression of STING, NLRP3, cleaved caspase-1, and GSDMD-NT in hippocampal neurons was up-regulated in P, PC and PV groups, and the expression of p-STING was significantly up-regulated in P and PV groups ( P<0.05). Compared with P group, the recognition index in the novel object recognition test was significantly increased, the number of crossing the original platform was reduced and the duration spent in the target quadrant was prolonged in the Morris water maze test, and the expression of p-STING, NLRP3, cleaved caspase-1, and GSDMD-NT in hippocampal neurons was down-regulated in PC group ( P<0.05). Compared with PC group, the recognition index in the novel object recognition test was significantly decreased, the number of crossing the original platform was reduced and the duration spent in the target quadrant was shortened in the Morris water maze test, and the expression of p-STING, NLRP3, cleaved caspase-1 and GSDMD-NT was up-regulated in PV group ( P<0.05). Conclusions:STING is involved in the development of POCD in aged mice, and the mechanism may be related to promotion of pyroptosis in hippocampal cells.

Objective:To present a set of clinically representative quality assurance (QA) test cases for stereotactic radiosurgery (SRT) plans of multiple intracranial metastases, in order to assess the plan quality and machine execution capabilities.Methods:Based on the clinical characteristics of multiple brain metastases, four groups of test cases with three target volumes (TVs), six TVs, nine TVs, and TVs near organs at risk (OARs) were designed. For these cases, SRT plans were developed, and plan quality was assessed using metrics including the Radiation Therapy Oncology Group conformality index (RTOG CI), gradient index (GI), homogeneity index (HI), and the volume of normal brain tissue receiving a dose of 24 Gy ( V24 Gy), which was defined as the volume enclosed by the 24 Gy isodose line around the Brain-PTV ( V24 Gy of Brain-PTV). Verification plans were generated for each test case, including the verification of point doses, planar doses (PD), and SRS MapCHECK (SMC) semiconductor matrix planar doses. Compared with the calculated result of the treatment planning system (TPS), the criteria for the γ analysis of planar doses were set at 1 mm/2% and 2 mm/2%. Results:For the four groups of test cases, the mean CI, GI, HI, and V24 Gy of Brain-PTV were 1.04±0.03, 3.79±0.40, 0.73±0.01 and (7.46±3.80) cm 3, respectively. The mean deviations of the point doses were 0.88%±0.98%, 1.47%±0.79%, 1.52%± 0.76%, and 1.17% ± 0.38%, respectively. The mean γ passing rates of the single fields for PDs were greater than 98% at 2 mm/2% and exceeding 96% at 1 mm/2%, and the mean γ pass rates of the SMC semiconductor matrix for PDs were 97.75% ± 2.31% and 99.33% ± 0.62%, at 1 mm/2% and 2 mm/2% respectively. Conclusions:The proposed QA test cases for SRT of multiple intracranial metastases allow for the effective assessments of the plan quality and machine execution capabilities and, thus, can assist various centers in clinical applications.

Objective:To investigate the therapeutic efficacy and disruptive effects of Meropenem（MEM）-loaded microbubbles（MBs）combined with ultrasound targeted microbubble destruction（UTMD）technology on Escherichia coli and its biofilm.Methods:MEM-MBs were prepared using the thin-film hydration method，and their characterization was assessed using a Zeta potential analyzer，with morphological observations conducted under an optical microscope. An in vitro biofilm model of periprosthetic joint infection（PJI）caused by Escherichia coli was constructed，and the morphology of the biofilm and the distribution of MEM-MBs in the bacterial biofilm were observed under a laser confocal microscope after staining the biofilm with SYTO59 staining and DIL staining for Microbubbles. The biofilm morphology and the distribution of MEM-MBs in bacterial biofilm were observed under laser confocal microscope. The biofilms were randomly divided into 5 groups using a random number table：control，Meropenem（MEM），MEM-MBs，UTMD，and MEM-MBs+UTMD，with 12 samples per group. After applying the respective interventions，scanning electron microscopy（SEM）and laser scanning confocal microscopy（LSCM）were employed to observe the effects on the morphology and structure of Escherichia coli and its biofilm. Crystal violet staining was utilized to determine and compare the biofilm density among groups using a microplate reader. LSCM was also used to observe the biofilm thickness，while both LSCM and spread plate counting were employed to assess bacterial viability differences across groups.Results:①MEM-MBs meeting the experimental requirements were successfully constructed.②A dense Escherichia coli biofilm visible under both the naked eye and LSCM was established，with a thickness of（10.61 ± 0.17）μm and a proportion of dead bacteria within the biofilm of（16.8 ± 0.8）%.③MEM-MBs were observed to penetrate into all layers of the biofilm using LSCM.④The results of crystal violet staining showed a decreasing trend in the biofilm density of the control group，the MEM group，the MEM-MBs group，the UTMD group，and the MEM-MBs+UTMD group. There was no significant difference between the MEM group and the MEM-MBs group（ P＞0.05），while there was a significant difference in biofilm density between the other groups，as revealed by pairwise comparison（all P＜0.05）.⑤UTMD technique and MEM-MBs+UTMD could significantly disrupt the biofilm of Escherichia coli. LSCM results showed that，compared to the control group，the thickness of the biofilm was reduced in all other groups，with only the UTMD group and the MEM-MBs+UTMD group showing an increase in porosity（both P＜0.05）. In comparison with the MEM group and the MEM-MBs group，the UTMD group showed an increase in porosity，while the MEM-MBs+UTMD group had a decrease in biofilm thickness and an increase in porosity（both P＜0.05）. Additionally，compared to the UTMD group，the MEM-MBs+UTMD group had a decrease in biofilm thickness and an increase in porosity（both P＜0.05），based on laser confocal microscopy results.⑥The results of the plate counting and LSCM showed that，compared with the control group，clump counts decreased，and the proportion of dead cells increased in the MEM group，the MEM-MBs group，and the MEM-MBs+UTMD group（all P＜0.05）. Compared with MEM group and MEM-MBs group，the clump counts of UTMD group increased，the proportion of dead cells decreased（all P＜0.05）；the clump counts of MEM-MBs+UTMD group decreased，and the proportion of dead cells increased（all P＜0.05）.Compared with UTMD group（all P＜0.05），the clump counts of MEM-MBs+UTMD group decreased，while the proportion of dead cells increased（all P＜0.05）.⑦The results of scanning electron microscopy revealed that the network structure of Escherichia coli was completely destroyed in the MEM-MBs+UTMD group. Conclusions:UTMD technology combined with MEM-MBs exerts a significant disruptive effect on the morphology and structure of Escherichia coli biofilm and significantly enhances bactericidal efficacy.

OBJECTIVES: This study aimed to observe the effects of initial periodontal therapy on the level of neutrophil extracellular traps (NETs) in gingival crevicular fluid (GCF) of patients with severe periodontitis and to analyze the factors related to the formation of NETs. METHODS: Thirty-one patients with stage Ⅲ-Ⅳ periodontitis were recruited. Clinical periodontal parameters, including plaque index (PLI), gingival index (GI), probing depth (PD), and clinical atta-chment loss (CAL), were recorded before and 6-8 weeks after initial periodontal therapy. Levels of NETs in GCF were detected by immunofluorescence staining. Quantities of total bacteria, Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actionomycetemcomitans) and Prevotella intermedia (P. intermedia)in unattached subgingival plaque were determined by real-time quantitative PCR, and levels of tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in GCF were explored by enzyme-linked immunosorbent assay. In addition, the correlations between the level of NETs and the above indicators were analyzed. RESULTS: After initial periodontal therapy, the level of NETs in GCF, PLI, GI, PD, and CAL; quantities of total bacteria, P. gingivalis, A. actinomycetemcomitans, and P. itermedia; and levels of IL-8 and TNF-α significantly decreased (P<0.05). We observed strong positive correlations between the level of NETs and PLI, GI, PD, CAL, the amount of total bacteria, P. gingivalis, TNF-α, and IL-8 (P<0.05). CONCLUSIONS Initial periodontal therapy might decrease the level of NETs in GCF from patients with severe periodontitis, which might be positively correlated with the quantities of P. gingivalis andthe levels of TNF-α and IL-8 in GCF.
Humans ; Gingival Crevicular Fluid ; Extracellular Traps/metabolism* ; Porphyromonas gingivalis/isolation & purification* ; Aggregatibacter actinomycetemcomitans/isolation & purification* ; Periodontitis/metabolism* ; Tumor Necrosis Factor-alpha/analysis* ; Prevotella intermedia/isolation & purification* ; Interleukin-8/analysis* ; Male ; Female ; Middle Aged ; Periodontal Index ; Adult

Objective:To establish a method of dual nueleic acid test strips for rapid detection of Bacillus cereus cytK and nhe toxin genes based on polymerase chain reaction(PCR)and colloidal gold technique,and to evaluate its specificity,sensitivity,repeatability and stability.Methods:Bacillus cereus DNA was extracted by boiling method.Specific primers were designed with Bacillus cereus cytK and nhe as the target genes.Clonal transformation was used to identify the PCR products.The optimal labeling amounts of colloidal gold-labeled streptavidin,quality control line(C line),cytK detection line(T1)and nhe detection line(T2)were determined.The nucleic acid test strips were assembled and its specificity,sensitivity,reproducibility and stability were evaluated.Results:The DNA concentration of Bacillus cereus was 248 mg·L-1,and the purities were 1.8-2.0.After cloning and plasmid sequencing,the similarities between the PCR products and the sequences of cytK and nhe registered in the GenBank database were 100％.Under the condition of pH 7.0,the optimal amount of streptavidin labeling per 200 μL of colloidal gold solution was 6.0 μL;the optimal marking amount was 2.00 g·L-1 for the quality control line(C line),0.550 g·L-1 for cytK gene detection line(T1)and 0.2 g·L-1 for nhe gene detection line(T2).In the specificity test,positive result on the test strips was seen only for Bacillus cereus,and no cross-reactivity was observed for Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa and Bacillus subtilis,which were consistent with the electrophoresis results.Sensitivity assay showed that even when DNA concentration was reduced to 10-2 mg·L-1,three bands(C line,T1 line and T2 line)could be observed,and the detection limit of the test strip was one-tenth of agarose gel electrophoresis(10-1 mg·L-1).The nucleic acid test strips were verified by different operators in different laboratories,and the results were consistent.The stability of the test strips was verified at the 6th,9th and 12th months,and the results showed good stability.Conclusion:The dual nucleic acid test strip method established in this study can simutaneously detect the cytK and nhe toxin genes of Bacillus cereus with high sensitivity and specificity,achieving short-term visual detection.