Background/Aims: Metabolic dysfunction-associated steatotic liver disease (MASLD) has become an increasingly important health challenge, with a substantial rise linked to changing lifestyles and global obesity. Ursolic acid, a natural pentacyclic triterpenoid, has been explored for its potential therapeutic effects. Given its multifunctional bioactive properties, this research further revealed the pharmacological mechanisms of ursolic acid on MASLD. Methods: Drug target chips and bioinformatics analysis were combined in this study to explore the potential therapeutic effects of ursolic acid on MASLD. Molecular docking simulations, surface plasmon resonance analyses, pull-down experiments, and co-immunoprecipitation assays were used to verify the direct interactions. Gene knockdown mice were generated, and high-fat diets were used to validate drug efficacy. Furthermore, initial CD4+ T cells were isolated and stimulated to demonstrate our findings. Results: In this study, the multifunctional extracellular matrix phosphorylated glycoprotein secreted phosphoprotein 1 (SPP1) was investigated, highlighting its capability to induce Th17 cell differentiation, amplifying inflammatory cascades, and subsequently promoting the evolution of MASLD. In addition, this study revealed that in addition to the canonical TGF-β/IL-6 cytokine pathway, SPP1 can directly interact with ITGB1 and CD44, orchestrating Th17 cell differentiation via their joint downstream ERK signaling pathway. Remarkably, ursolic acid intervention notably suppressed the protein activity of SPP1, suggesting a promising avenue for ameliorating the immunoinflammatory trajectory in MASLD progression. Conclusions Ursolic acid could improve immune inflammation in MASLD by modulating SPP1-mediated Th17 cell differentiation via the ERK signaling pathway, which is orchestrated jointly by ITGB1 and CD44, emerging as a linchpin in this molecular cascade.

Objective:To compare proximal humerus internal locking system (PHILOS) and Multiloc intramedullary nail in the treatment of proximal humerus fracture-anterior dislocation.Methods:A retrospective study was performed to analyze the data of 33 patients with proximal humerus fracture-anterior dislocation who had been treated by open reduction and internal fixation from June 2015 to April 2021 at Department of Upper Limbs, Zhengzhou Orthopaedic Hospital. According to methods of internal fixation, the patients were divided into an extramedullary group and an intramedullary group. In the extramedullary group of 18 cases subjected to internal fixation with PHILOS, there were 8 males and 10 females with an age of (53.3 ± 10.6) years, and 1 2-part fracture, 15 3-part fractures and 2 4-part fractures by the Neer classification. In the intramedullary group of 15 cases subjected to internal fixation with Multiloc intramedullary nail, there were 8 males and 7 females with an age of (51.5 ± 11.2) years, and 14 3-part fractures and 1 4-part fracture by the Neer classification. The 2 groups were compared in terms of incision length, operation time, intraoperative blood loss, postoperative complications, and visual analog scale (VAS), range of shoulder motion, and Constant-Murley score at postoperative 12 months.Results:The 2 groups were comparable due to insignificant differences in their preoperative general data ( P>0.05). All patients were followed up for (20.8 ± 4.7) months. The incision length in the intramedullary group [(11.6 ± 1.7) cm] was significantly shorter than that in the extramedullary group [(17.6 ± 2.0) cm], and the intraoperative blood loss in the former [(106.7 ± 34.4) mL] was significantly lower than that in the latter [(151.7 ± 45.7) mL] ( P<0.05). The VAS scores at 1 week and 1 month after surgery [2.0 (2.0, 3.0) and 0.0 (0.0, 1.0) respectively] in the intramedullary group were significantly lower than those in the extramedullary group [3.0 (3.0, 3.3) and 1.0 (0.0, 1.3) respectively] ( P<0.05). The external rotation of the shoulder at the last follow-up in the intramedullary group (65.3° ± 15.5°) was significantly larger than that in the extramedullary group (50.6° ± 13.9°) ( P<0.05). There were no significant differences in operation time, incidence of postoperative complications, VAS score at 12 months after operation, Constant-Murley score or range of shoulder motion at the last follow-up between the 2 groups ( P>0.05). Conclusions:In the treatment of proximal humerus fracture-anterior dislocation, open reduction and internal fixation with both PHILOS and Multiloc intramedullary nail can result in a favorable prognosis when the fracture-dislocation is well reduced and fixated. However, the Multiloc intramedullary nail may lead to better early pain relief, less surgical invasion, and better functional recovery of the external rotation of the shoulder.

Objective:To evaluate the role of monocyte-derived macrophages (Mo-Mφ) in postoperative cognitive dysfunction (POCD) induced by cardiopulmonary bypass (CPB) in rats.Methods:Forty SPF healthy adult Sprague-Dawley rats, weighing 350-400 g, aged 11-12 weeks, were divided into 4 groups ( n = 10 each) using a random number table method: sham operation group (S group), CPB group (C group), CPB plus PBS liposome group (P group), and CPB plus clodronate liposome group (L group). In C, P, and L groups, CPB was performed for 60 min, the equal volume of normal saline, PBS liposomes 4 μl/g and clodronate liposomes 4 μl were injected via the tail vein at 48 and 24 h before CPB, respectively.Blood was collected from the saphenous vein before CPB for determination of the clearance rate of Mo-Mφ by flow cytometry.Cognitive function was assessed using the Morris water maze test at 7 days after CPB.Blood was collected from the abdominal aortic vein, and the levels of serum inflammatory factors (interleukin-6 [IL-6], tumor necrosis factor-alpha [TNF-α], and interleukin-1beta [IL-1β]) and brain injury markers (S100β protein and neuron-specific enolase [NSE]) were measured by enzyme-linked immunosorbent assay. Results:Compared with group S, the serum IL-1β, IL-6 and TNF-α concentrations were significantly increased, the serum S100β protein and NSE concentrations were increased, the escape latency was prolonged, and the number of crossing the platform was decreased in C, P, and L groups ( P<0.05). Compared with group C, the clearance rate of blood-derived monocytes was significantly decreased, the serum S100β concentrations were increased, the escape latency was prolonged, and the number of crossing the platform was decreased ( P<0.05), and no significant change was found the parameters mentioned above in group P ( P>0.05). Conclusion:Mo-Mφ infiltration is the endogenous protective mechanism of CPB-induced POCD in the rats.

OBJECTIVE: To investigate serum levels of long non-coding RNA (lncRNA) TUSC7 in patients with esophageal squamous cell carcinoma (ESCC), its association with clinicopathological parameters and its role in promoting tumor metastasis and invasion. METHODS: Serum samples were collected from 60 patients with ESCC admitted between January, 2017 and May, 2019, with 60 age- and gender-matched healthy subjects as the control group. Serum level of TUSC7 in ESCC patients and its expression in 4 ESCC cell lines was detected with RT-qPCR. The association of serum TUSC7 level with the clinicopathological features of the patients was analyzed. KYSE-30 cell models with TUSC7 overexpression or knockdown were established, and the proliferation of the cells was examined with MTT assay and their migration and invasion were assessed using wound healing and Transwell assays. Western blotting was used to detect the cellular expressions of the proteins associated with epithelial-mesenchymal transition (EMT). RESULTS: The patients with ESCC had significantly lower serum TUSC7 level than the healthy control subjects ( < 0.05). The ESCC cell lines also expressed lower levels of TUSC7 than normal cells ( < 0.05). Serum TUSC7 level was negatively correlated with tumor staging, lymph node metastasis and infiltration ( < 0.05) but was not significantly correlated with other clinicopathological parameters in ESCC patients. In the cell experiment, overexpression of TUSC7 in KYSE-30 cells significantly inhibited cell migration and invasion ( < 0.05), enhanced the expression of the EMT marker protein E-cadherin and lowered the expressions of N-cadherin, Vimentin and MMP9 ( < 0.05); knocking down TUSC7 in the cells produced the opposite effects. CONCLUSIONS The down-regulation of TUSC7 expression in the serum of ESCC patients and in ESCC cell lines is associated with the metastasis of ESCC and promotes tumor cell migration and invasion by promoting EMT, indicating the potential of serum TUSC7 level as a molecular marker for diagnosis, treatment and metastasis monitoring of ESCC.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms ; genetics ; Esophageal Squamous Cell Carcinoma ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasm Invasiveness ; RNA, Long Noncoding ; genetics

Objective To investigate the expressions of regulatory B cells and related cytokines in patients with Guillain-Barre syndrome (GBS).MethodsForty-four patients with GBS admitted to our hospital from October 2018 to June 2019 were enrolled as GBS group; 44 healthy subjects accepted physical examination in our hospital at the same period were selected as control group. Flow cytometry was used to determine the proportions of CD19+CD24hiCD38hi regulatory B cells and CD19+CD24hiCD27+ regulatory B cells in CD19+ lymphocytes in peripheral blood mononuclear cells of the two groups. ELISA was employed to detect the serum contents of interleukin (IL)-10, IL-35 and transforming growth factor (TGF)-β1 of the two groups.ResultsAs compared with those in the control group, the proportions of CD19+CD24hiCD38hi regulatory B cells and CD19+CD24hiCD27+ regulatory B cells in CD19+B lymphocytes of the GBS group were significantly decreased (P<0.05). As compared with the control group, the GBS group had significantly decreased IL-10 and IL-35 levels (P<0.05), and obviously increased TGF-β1 content without statistical difference (P>0.05).ConclusionRegulatory B cells play a role in the pathogenesis of GBS through IL-10 and IL-35 cytokine pathways.

OBJECTIVEThe aim of this study was to investigate the use of the lesion-specific endonucleases-modified comet assay for analysis of DNA oxidation in cell lines.

METHODSDNA breaks and oxidative damage were evaluated by normal alkaline and formamidopyrimidine-DNA-glycosylase (FPG) modified comet assays. Cytotoxicity were assessed by MTT method. The human bronchial epithelial cell (16HBE) were treated with benzo (a) pyrene (B(a)P), methyl methanesulfonate (MMS), colchicine (COL) and vincristine (VCR) respectively, and the dose is 20 µmol/L, 25 mg/ml, 5 mg/L and 0.5 mg/L for 24 h, respectively. Oxidative damage was also detected by levels of reactive oxygen species in treated cells.

RESULTSFour genotoxicants give higher cytotoxicity and no significant changes on parameters of comet assay treated by enzyme buffer. Cell survival rate were (59.69 ± 2.60) %, (54.33 ± 2.81) %, (53.11 ± 4.00) %, (51.43 ± 3.92) % in four groups, respectively. There was the direct DNA damage induced by test genotoxicants presented by tail length, Olive tail moment (TM) and tail DNA (%) in the comet assay. The presence of FPG in the assays increased DNA migration in treated groups when compared to those without it, and the difference was statistically significant which indicated that the clastogen and aneugen could induce oxidative damage in DNA strand. In the three parameters, the Olive TM was changed most obviously after genotoxicants treatment. In the contrast group, the Olive TM of B(a) P,MMS, COL,VCR in the contrast groups were 22.99 ± 17.33, 31.65 ± 18.86, 19.86 ± 9.56 and 17.02 ± 9.39, respectively, after dealing with the FPG, the Olive TM were 34.50 ± 17.29, 43.80 ± 10.06, 33.10 ± 12.38, 28.60 ± 10.53, increased by 58.94%, 38.48%, 66.86% and 68.21%, respectively (t value was 3.91, 3.89, 6.66 and 3.87, respectively, and all P < 0.05), and the correlation between Olive TM and reactive oxygen species was better than other parameters (r = 0.77, P < 0.05).

CONCLUSIONThis study indicates that FPG-comet assay appears more specific for detecting oxidative DNA damage induced by genotoxicants exposure, and the application of comet assay will be expanded. The endonuclease modified comet assay will be used widely in the toxicology and molecular epidemiology study.

Cell Line ; Comet Assay ; methods ; DNA Damage ; Endonucleases ; Humans ; Mutagens ; toxicity ; Oxidation-Reduction ; Oxidative Stress ; Reactive Oxygen Species ; metabolism

ObjectiveTo discuss the application of neurophysiological monitoring (NEPM), intraoperative color Doppler ultrasonography, fluorescein angiography and neuroendoscope in clinical effects of intracranial giant aneurysm microsurgey. MethodsTo retrospectively review the clinical data of 17 intracranial giant aneurysm. Pre-operative imaging were used, including 3D- CTA, MRI and DSA, to make dectection and delineation of the aneurysm.The NEPM to evaluate the nerve function,assess the qualitative and quantitative flow rate of aneurysm and surrounding blood vessels by Doppler ultrasonography and fluorescein angiography,and reveal opography of aneurysm,protect the considerable perf.vessels and nerves by neuroendoscope.Operative techniques were used including parent artery control,aneurysm neck forming,aneurysm decompression and resection,obliteration of aneurysm with multiple clips and vasospasm protection.Results Seventeen cases of giant aneurysms were clipped successfully under muti-technology, follow-up demonstrated excellent neurological outcomes in 15 cases, one case had mild disability, one case had severe disability, no dead cases. DSA showed clipping completely, parent artery clear, and long-term follow-up was still in progress.Conclusion Multi-technology combined microsurgical techniques which can effective improve the outcomes of intracranial giant aneurysms.

Objective To investigate the effects of glycine on the expression of HSP70 and TNF-α mRNA in the liver tissue of rats with traumatic shock and explore the protective mechanism of glycine a-gainst secondary liver injury after traumatic shock. Methods The traumatic shock model was established and 120 Wistar rats were divided randomly into three groups: treatment group, shock group and control group. At the beginning of resuscitation, the rats in the treatment were injected with 0.5 ml isotonic saline containing 100 mg/kg glycine, those rats in the shock group were injected only with 0.5 ml isotonic saline. The rats in three groups were killed at 3, 6, 12, 24 and 48 hours after resuscitation respectively. The ex-pression of HSP70 and TNF-α mRNA in the liver tissue were detected by RT-PCR, pathological changes were observed and serum ALT and AST were measured. Results The expressions of HSP70 and TNF-α mRNA in the liver tissue of rats in the shock group began to increase at 3 hours and both reached the peak value at 6 hours after resuscitation, but the expression of HSP70 mRNA in the treatment group reached the peak value at 12 hours after resuscitation. Compared with the control group, the expression of HSP70 mR-NA in the treatment group increased significantly and that of TNF-α mRNA decreased siganicantly, serum ALT and AST decreased and pathological damage was relieved significantly (all P < 0.05). Conclusion By enhancing the expression of HSP70 mRNA and decreasing the expression of TNF-α mRNA, glycine may play a protective role against the secondary damage of liver after traumatic shock.