Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

Background/Aims: Metabolic dysfunction–associated steatotic liver disease (MASLD) is a chronic liver disease characterized by hepatic steatosis. Ubiquitin-specific protease 29 (USP29) plays pivotal roles in hepatic ischemiareperfusion injury and hepatocellular carcinoma, but its role in MASLD remains unexplored. Therefore, the aim of this study was to reveal the effects and underlying mechanisms of USP29 in MASLD progression. Methods: USP29 expression was assessed in liver samples from MASLD patients and mice. The role and molecular mechanism of USP29 in MASLD were assessed in high-fat diet-fed and high-fat/high-cholesterol diet-fed mice and palmitic acid and oleic acid treated hepatocytes. Results: USP29 protein levels were significantly reduced in mice and humans with MASLD. Hepatic steatosis, inflammation and fibrosis were significantly exacerbated by USP29 deletion and relieved by USP29 overexpression. Mechanistically, USP29 significantly activated the expression of genes related to fatty acid β-oxidation (FAO) under metabolic stimulation, directly interacted with long-chain acyl-CoA synthase 5 (ACSL5) and repressed ACSL5 degradation by increasing ACSL5 K48-linked deubiquitination. Moreover, the effect of USP29 on hepatocyte lipid accumulation and MASLD was dependent on ACSL5. Conclusions USP29 functions as a novel negative regulator of MASLD by stabilizing ACSL5 to promote FAO. The activation of the USP29-ACSL5 axis may represent a potential therapeutic strategy for MASLD.

ObjectiveTo explore the application value of probe-based confocal laser endomicroscopy (pCLE) in rapidly detecting and evaluating the morphological characteristics of digestive tract tissues in mice. MethodsTwelve male SPF Kunming mice aged 6 weeks were randomly divided into two groups. Six mice were subjected to gastric gavage with 52% Red Star Erguotou to establish the model, and six were given saline by gastric gavage as a control. After 28 days of modeling, 3 mice were randomly selected from each group. After deep anesthesia induced by inhalation of 3% isoflurane, the mice were sacrificed by cervical dislocation. The stomach, duodenum, jejunum, and rectum tissues were excised and immersed in 1% fluorescein sodium solution for staining. The microstructure of the mucosal surface of each tissue was observed using pCLE. The remaining mice in the model group and the control group were deeply anesthetized by inhaling 3% isoflurane, then cardiac perfusion was performed successively with saline and 4% paraformaldehyde. The stomach, duodenum, jejunum, and rectum tissues were excised for dehydration, section and hematoxylin-eosin (HE) staining, and the morphological changes of the tissues were observed under a microscope. ResultsUnder pCLE imaging, fluorescence staining on the surface of the gastrointestinal mucosa was uniform in the control group; the morphology of gastric pits, intestinal villi, and intestinal crypts was intact, arranged compactly, and had distinct boundaries. In the model group, the gastrointestinal mucosa exhibited mucosal swelling and deformation, with uneven fluorescence staining and fluorescein leakage. Furthermore, some tissues showed defects or cell shedding, and the boundaries between adjacent characteristic structures (e.g., gastric pits, intestinal crypts) were blurred. HE staining showed that the gastrointestinal tissue structure of the control group mice was normal and well-organized, with no structural defects. Moreover, submucosal glands were uniform in size, with no hyperplasia observed, and no obvious inflammatory cell infiltration. In the model group, some gastrointestinal mucosal structures were defective and sparsely arranged; submucosal glands showed atrophy, accompanied by obvious inflammatory cell infiltration. The histological characteristics detected by pCLE were consistent with those of HE staining. ConclusionpCLE can be used to obtain rapid, real-time, large-scale, and high-resolution microscopic imaging of the gastrointestinal mucosa, realistically and comprehensively displaying its physiological and microstructural characteristics. It shows promising prospects and practical utility in the histological evaluation of digestive system injuries in small animals.

OBJECTIVE: To compare the aesthetic effects of anterior maxilla clockwise rotation combined with segmental Le Fort Ⅰ osteotomy and whole maxilla clockwise rotation on improving paranasal concavity in patients with Class Ⅲ maxillofacial deformity. METHODS: A non-randomized controlled trial was designed, and 21 patients diagnosed with skeletal Class Ⅲ maxillofacial deformity were included. In the study, 11 patients in the test group were treated by segmental Le Fort Ⅰ osteotomy combined with anterior maxilla clockwise rotation, and 10 patients in the control group were treated by whole maxilla clockwise rotation. The CBCT and 3D photography of preoperative (T0), 2 weeks postoperative (T1), and 6 months postoperative (T2) were collected respectively, and the three-dimensional cephalometry was carried out. The differences of specific parameters between the two groups were compared by independent sample t-test, including saggital displacement of the cheek mass point (CK) and subalare point (SA), nasolabial angle, occlusal plane angle and labial inclination angle of the upper incisor. RESULTS: There were no significant differences of the parameters on T0 between the two groups. The average sagittal displacement of the upper incisors of the test group was (-0.71±1.67) mm and smaller than that of control group [(2.26±1.68) mm], t=-4.052, P < 0.05. The average angle of the occlusal plane clockwise rotation of the test group was 1.46°±2.38° and smaller than that of the control group (4.31°±1.83°), t=-3.047, P < 0.05. The angle of anterior maxilla clockwise rotation was 11.73°±2.81° during the surgery. The average saggital displacement of the paranasal soft tissue landmarks of the test group from T0 to T2 was larger than that of the control group [CK point, (4.96±1.18) mm vs. (2.01± 1.50) mm, P < 0.05;SA point, (5.19±1.17) mm vs. (2.69±1.45) mm, P < 0.05]. The labial inclination angle of the upper incisor of the test group was 112.15°±5.40° in T2 and significantly smaller than that of the control group (122.38°±8.83°), t=-3.237, P < 0.05. The nasolabial angle of the test group was 106.54°±12.82° in T2 and significantly larger than that of the control group (93.90°±12.46°), t=2.288, P < 0.05. CONCLUSION Compared with whole maxilla clockwise rotation, anterior maxilla clockwise rotation combined with segmental Le Fort Ⅰ osteotomy can increase the saggital displacement of the paranasal soft tissue, correct labial inclination of the upper incisors and the acute naso-labial angle and better improve the paranasal aesthetic defects in patients with Class Ⅲ maxillofacial deformity with less changing on the saggital orientation of the upper incisors and the occlusal plane angle.
Humans ; Maxilla/surgery* ; Female ; Male ; Osteotomy, Le Fort/methods* ; Malocclusion, Angle Class III/surgery* ; Adult ; Cephalometry ; Young Adult ; Rotation ; Adolescent

OBJECTIVE To screen and quantitatively analyze differential quality markers （Q-Marker） in Zingiber officinale mixed and triturated with Schisandra chinensis （ZMTS） before and after processing. METHODS HPLC fingerprints of before processing [Z. officinale complicated with S. chinensis （ZWS）] and after processing （ZMTS） （10 batches each） were established. The differences of Q-Markers before and after processing were screened by the chemical pattern recognition method and Q-Marker “five principles”， and the contents were determined. RESULTS A total of 14 common peaks were identified in the fingerprints of ZWS， 22 common peaks were identified in the fingerprints of ZMTS， and 8 components were identified. Differential Q-Marker were screened by chemical pattern recognition and Q-Marker “five principles”， i. e. 6-gingerol， schisandrol A schisandrol B， 8-gingerol， 10-gingerol， schisandrin A， schisandrin B， schizandrin C. The average contents of the 8 differential Q-Markers in ZMTS were 229.46， 244.48， 39.96， 44.12， 61.17， 47.82， 100.11 and 9.70 μg/g， respectively. The average contents of the 4 differential Q-Markers （6-gingerol， schisandrol A， schisandrol B， 8-gingerol） in ZWS were 112.58， 19.01， 26.74 and 5.98 μg/g， respectively. CONCLUSIONS In this study， the differential Q-Markers before and after ZMTS processing are screened. The contents of the Q-Markers in ZMTS after processing are higher than those before processing.

Objective Studies on the expression and location of zinc finger protein A20 (A20) and connective tissue growth factor (CTGF) in liver tissues of patients with chronic hepatitis B were conducted, and the relationship between them and liver fibrosis was determined by FibroScan. Methods Studies on A20 and CTGF in liver tissues of 160 patients with chronic hepatitis B were conducted in accordance with the stage of pathological fibrosis and inflammation of the liver, and quantitative immunohistochemistry test was conducted, and statistical analysis was conducted by FibroScan. Results The expressions of A20 and CTGF in liver tissues increased with the aggravation of liver pathological fibrosis and inflammation, and there were significant differences between each stage and the control group (P<0.05), and there were significant differences between adjacent groups (P<0.05). Studies have shown that FibroScan increases along with pathological fibrosis and inflammation in the liver. There are significant differences between the stage and the control group (P<0.05), and no significant differences between the adjacent groups (P>0.05). There was positive correlation between liver A20 and CTGF, r=0.796 (P<0.05). Conclusions In patients with chronic hepatitis B, A20, CTGF and FibroScan are positively correlated with the degree of liver fibrosis, and A20 and CTGF are also positively correlated with the degree of liver inflammation, which can be used as indicators to evaluate the degree of liver inflammation and fibrosis, and further guide the anti-inflammatory and anti-fibrosis treatment of patients.

Objective:To establish and assess the precision of pre-surgical condyle position planning using mandibular movement trajectory data for orthognathic surgery.Methods:Skull data from large-field cone beam computed tomography(CBCT)and dental oral scan data were imported into IVSPlan 1.0.25 software for 3D reconstruction and fusion,creating 3D models of the maxilla and mandible.Trajectory da-ta of mandibular movement were collected using a mandibular motion recorder,and the data were inte-grated with the jaw models within the software.Subsequently,three-dimensional trajectories of the con-dyle were obtained through matrix transformations,rendering them visually accessible.A senior oral and maxillofacial surgeon with experience in both diagnosis and treatment of temporomandibular joint disease and orthognathic surgery selected the appropriate condyle position using the condyle movement trajectory interface.During surgical design,the mobile mandibular proximal segment was positioned accordingly.Routine orthognathic surgical planning was completed by determining the location of the mandibular distal segment,which was based on occlusal relationships with maxilla and facial aesthetics.A virtual mandible model was created by integrating data from the proximal and distal segment bone.Subsequently,a solid model was generated through rapid prototyping.The titanium plate was pre-shaped on the mandibular model,and the screw hole positions were determined to design a condylar positioning guide device.In accordance with the surgical plan,orthognathic surgery was performed,involving mandibular bilateral sagittal split ramus osteotomy(SSRO).The distal segment of the mandible was correctly aligned inter-maxillary,while the proximal bone segment was positioned using the condylar positioning guide device and the pre-shaped titanium plate.The accuracy of this procedure was assessed in a study involving 10 patients with skeletal class Ⅱ malocclusion.Preoperative condyle location planning and intraoperative po-sitioning were executed using the aforementioned techniques.CBCT data were collected both before the surgery and 2 weeks after the procedure,and the root mean square(RMS)distance between the preope-rative design position and the actual postoperative condyle position was analyzed.Results:The RMS of the condyle surface distance measured was(1.59±0.36)mm(95％CI:1.35-1.70 mm).This value was found to be significantly less than 2 mm threshold recommended by the expert consensus(P＜0.05).Conclusion:The mandibular trajectory may play a guiding role in determining the position of the mandibular proximal segment including the condyle in the orthognathic surgery.Through the use of a con-dylar positioning guide device and pre-shaped titanium plates,the condyle positioning can be personalized and customized with clinically acceptable accuracy.

ObjectiveTo compare the effects of different doses and withdrawal time of 4-vinylcyclohexene diepoxide （VCD） on the reproductive endocrine levels of female rats， and to explore the effective， stable， and safe dosage of VCD for constructing a premature ovarian insufficiency （POI） rat model. MethodSD rats with regular estrous cycles were randomly divided into three groups： blank group， low-dose VCD group （80 mg·kg^-1·d^-1）， and high-dose VCD group （160 mg·kg^-1·d^-1）， with 24 rats in each group. After drug intervention， samples were collected on the 15th day （D15） and the 45th day （D45） after intervention. The general condition， rate of estrous cycle disturbance， serum hormone levels， ovarian histomorphology， follicle count， pregnancy outcome， and the protein and mRNA expression of transforming growth factor （TGF）-β and Smad2/3 were assessed. ResultCompared with the blank group， the low-dose VCD group showed no significant differences in the rate of estrous cycle disturbance or serum follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E₂） levels. Ovarian tissue was damaged. Specifically， the number of primordial and primary follicles decreased on D15 （P<0.01）， and the number of secondary follicles （P<0.01） and antral follicles （P<0.05） further decreased on D45. The litter number decreased on D15 （P<0.05）， but there was no significant difference on D45. Furthermore， TGF-β protein levels increased on D15 （P<0.05） and D45 （P<0.01）. The Smad2/3 levels increased on D45 （P<0.01）， and TGF-β and Smad2/3 mRNA levels increased on D45 （P<0.05）. Compared with the results in the blank group， the disturbance rate of the estrous cycle increased on D45 in the high-dose VCD group （P<0.01）. The serum of FSH and LH increased （P<0.01）， while E₂ decreased （P<0.05）. Ovarian tissue was damaged， and the downward trend of follicles at all levels was similar to that in the low-dose VCD group. The litter number significantly decreased on D15 and D45. TGF-β and Smad2/3 protein levels increased （P<0.05，P<0.01）， and TGF-β mRNA increased on D45 （P<0.05）. ConclusionHigh-dose VCD is an ideal method for constructing a POI rat model， being effective， stable， and safe.