Metabolic dysfunction-associated steatotic liver disease(MASLD)is a chronic liver condition intricately linked to metabolic abnormalities such as obesity,type 2 diabetes,dyslipidemia,and hypertension.The global prevalence of MASLD continues to rise,posing a significant public health challenge.The pathogenesis of MASLD is multifactorial,with the"multiple-hit"hypothesis suggesting that hepatic lipid accumulation,insulin resistance,oxidative stress,gut microbiota dysbiosis,and genetic factors collectively drive disease progression.Currently,clinical management primarily relies on lifestyle interventions;however,there is a lack of targeted pharmacological interventions,and there is an urgent need to investigate novel adjunctive therapeutic strategies.In recent years,probiotics have demonstrated potential value in MASLD treatment due to their capacity to modulate gut microbiota,enhance insulin sensitivity,and reduce liver inflammation.This review systematically examines the pathogenesis of MASLD and the limitations of existing therapeutic approaches,synthesizing the latest evidence of probiotics-assisted therapy for MASLD from the perspectives of animal studies and clinical trials.By analyzing the target mechanisms and molecular pathways of different strains(e.g.,Bifidobacterium,Lactobacillus),this review explores the translational potential of probiotics in MASLD treatment,aiming to provide a theoretical foundation and future research directions.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

The oral glucose growth hormone suppression test is commonly used in the clinical diagnosis of acromegaly, but its results can be influenced by a variety of factors. This case report discusses a patient with a pituitary tumor and concurrent liver cirrhosis, highlighting the complexities in interpreting test results under such conditions. The patient, a 54-year-old male, presented with blurred vision as his primary complaint. Notably, the physical examination revealed no changes in facial features, no enlargement of hands or feet, and no other symptoms typically associated with acromegaly, which might otherwise suggest excessive growth hormone activity. Magnetic Resonance Imaging (MRI) of the pituitary gland indicated that the gland was within normal size parameters, but a small low-intensity lesion mea-suring approximately 3 mm×2 mm identified. This finding was consistent with a pituitary microadenoma. The patient's fasting growth hormone levels were significantly elevated at 8.470 μg/L, compared with the normal range of less than 2.47 μg/L. Conversely, fasting insulin-like growth factor-1 (IGF-1) levels were notably low, recorded at 41 and 52 μg/L, whereas the normal range for a person of his age was between 87 and 234 μg/L. Other pituitary hormones, including those regulating the thyroid, adrenal cortex, and sex hormones, were found to be within normal ranges. Despite this, during the glucose growth hormone suppression test, an abnormal elevation of growth hormone was observed. To investigate further, the patient was administered branched-chain amino acids, and the suppression test was repeated. However, the abnormal elevation of growth hormone persisted, indicating a failure to normalize the response. Given the patient's lack of clinical signs typically associated with elevated growth hormone secretion, the history of liver cirrhosis became a significant consideration. The disparity between elevated growth hormone levels and reduced IGF-1 levels suggested that the pituitary lesion was a non-functional adenoma rather than a source of excess hormone production. Consequently, it was concluded that the abnormal response of growth hormone to the glucose suppression test was likely related to the patient's liver cirrhosis. In addition to chronic liver disease, various other conditions could influence the results of the oral glucose tolerance growth hormone suppression test. According to the literature, factors such as puberty, diabetes, anorexia nervosa, and protein malnutrition could also affect test outcomes. These conditions could cause similar abnormalities in growth hormone dynamics, complicating the diagnosis. Therefore, clinicians must be vigilant and consider these potential influences when interpreting test results.For an accurate diagnosis of acromegaly, it is essential to combine clinical symptoms, detailed medical history, and imaging studies. The presence of conditions like liver cirrhosis should prompt careful interpretation of the test results, ensuring that other contributing factors are not overlooked. This comprehensive approach is crucial to avoid misdiagnosis and to ensure that appropriate treatment strategies are implemented based on a thorough understanding of the patient's overall health status.
Humans ; Male ; Middle Aged ; Pituitary Neoplasms/blood* ; Liver Cirrhosis/blood* ; Adenoma/blood* ; Human Growth Hormone/blood* ; Insulin-Like Growth Factor I/metabolism* ; Acromegaly/etiology* ; Magnetic Resonance Imaging

A 20-year old female patient diagnosed with right distal tibial giant cell tumor underwent a surgery of resection of distal tibial giant cell tumor, residual cavity liquid nitrogen and electrocauterization inactivation and local arthroplasty on March 17, 2023. Preoperatively a life-size distal tibia model was 3D printed using polylactic acid (PLA) material based on the CT data of patient's distal tibia. Tumor resection was simulated on the model, preserving the surrounding normal bone and articular cartilage unaffected by the tumor. The residual cavity was filled with bone cement and the distal tibial articular surface was shaped using the talar articular surface as a template. The 3D CT data of bone cement was collected and reconstructed. The irregular bone and cartilage defect data were trimmed to form a regular arc shape, which was used as the data for fabricating local arthroplasty prosthesis. The local arthroplasty prosthesis composed of a titanium base and a VE polyethylene liner was 3D printed. During the operation, the test models of titanium alloy base and VE polyethylene liner were used to test the matching degree with the bone and cartilage defect. Minor adjustments were made by removing a portion of the lateral wall of the residual cavity and modifying the base height to achieve proper alignment of the distal tibial articular surface with the talar surface. After confirming a satisfactory fit, the local arthroplasty prosthesis was implanted. Intraoperative fluoroscopic confirmed accurate placement of the prosthesis, good anatomical match with the defect, and restoration of the joint line. The postoperative follow-up was conducted at 2, 4, 12, 20, 48, 72 and 92 weeks. Wound healing was closely monitored, along with radiologic assessment for prosthesis bone ingrowth and local tumor recurrence. Functional evaluations were performed using the AOFAS and Kofoed scoring systems. Postoperatively, the patient experienced plantar numbness and sensory disturbance, which gradually resolved after three weeks. Assisted ambulation began at two weeks postoperatively, and the patient resumed a normal gait by 12 weeks. The Mayo ankle arthroplasty evaluation criteria at postoperative at 48 weeks were excellent. The AOFAS score and Kofoed score were 97 points and 94 points respectively, indicating excellent functional outcomes. Postoperative X-ray indicated that no bone ingrowth was observed at 2 weeks and 4 weeks after the operation, minor ingrowth at 12 weeks postoperatively, significant bone ingrowth at 20 weeks, and complete osseointegration by 48 to 64 weeks. Postoperative CT imaging at 92 weeks confirmed full prosthesis osseointegration, while MRI at 72 weeks showed no evidence of tumor recurrence.

Objective:To evaluate the role of the CC chemokine ligand 2/CC chemokine receptor 2 (CCL2/CCR2) signaling pathway in electroacupuncture (EA)-induced reduction of spinal cord injury (SCI) in rats.Methods:Sixty clean-grade healthy adult female Sprague-Dawley rats, weighing 210-250 g, were divided into 5 groups ( n=12 each) using a random number table method: sham operation group (group S), group SCI, SCI+ Anti-CCL2 group (group SCI+ A), SCI+ EA group (group SCI+ EA), and spinal cord injury+ EA+ rCCL2 group (group SCI+ EA+ R). The SCI model was established using the Allen method in anesthetized animals. Group S only underwent spinous process and laminectomy without damaging the spinal cord. In SCI+ A group, CCL2 neutralizing antibody 50 μg/kg was intrathecally injected at 0, 3 and 6 days after successful development of the SCI model. On the 7th day after the successful development of the SCI model, Jiaji, Dazhui and Mingmen acupoints were stimulated with a depth of 2 mm, voltage of 12-15 mV and frequency of 2 Hz for 30 min once a day for 7 consecutive days in SCI+ EA group. In SCI+ EA+ R group, recombinant rat CCL2 2.5 μg/kg was intrathecally injected at the site of injury at 0, 3 and 6 days after successful development of the SCI model, and the remaining treatments were similar to those in SCI+ EA group. At 1 day before developing the model, 0, 3, 7, 14, 21 and 28 days after developing the model, the mechanical paw withdraw threshold (MWT) and thermal paw withdrawal latency (TWL) were measured, and the motor function was assessed by BBB score. The rats were sacrificed after the final behavioral testing, and their spinal cord tissues were obtained for determination of the expression of CCL2 and CCR2 protein and mRNA (by Western blot or quantitative real-time polymerase chain reaction), the expression of GFAP (by immunofluorescence), contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay) and for examination of the pathological changes (using HE staining). Results:Compared with S group, the MWT and BBB scores were significantly decreased and the TWL was shortened at each time point after developing the model, the expression of CCL2 and CCR2 protein and mRNA and GFAP was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased in SCI group ( P<0.05). Compared with SCI group, the MWT and BBB scores were significantly increased, and the TWL was prolonged at 7 days after developing the model in SCI+ A group, the MWT and BBB scores were significantly increased, and the TWL was prolonged at 14 days after developing the model in SCI+ EA group, and the expression of CCL2 and CCR2 protein and mRNA and GFAP was significantly down-regulated, and the contents of TNF-α, IL-1β and IL-6 were decreased in SCI+ A and SCI+ EA groups ( P<0.05). Compared with SCI+ EA group, the MWT and BBB scores were significantly decreased at 14 days after developing the model, the TWL was shortened, the expression of CCL2 and CCR2 protein and mRNA and GFAP was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased in SCI+ EA+ R group ( P<0.05). Compared with SCI+ A and SCI+ EA groups, the histopathological injury were significantly attenuated in SCI group, and the histopathological injury was aggravated in SCI+ EA+ R group. Conclusions:The CCL2/CCR2 signaling pathway is involved in the process by which EA reduces SCI, and the mechanism is related to the inhibition of astrocyte activation, thereby reducing the inflammatory response.

Objective:To evaluate the effect of electroacupuncture (EA) on ionotropic purinergic receptor 4 (P2X4R)/nuclear factor-kappa B (NF-κB) signaling pathway during spinal cord injury (SCI) in rats.Methods:Thirty-six clean-grade healthy adult female Sprague-Dawley rats, weighing 210-250 g, were divided into 3 groups ( n=12 each) using a random number table method: sham surgery group (S group), SCI group, and SCI+ EA treatment group (SCI+ EA group). The SCI model was established by the Allen′s method in anesthetized animals. In group S, only the spinous processes and vertebral laminae were resected, but the spinal cord was not injured. On the 7th day after developing the model, EA of Jiaji, Dazhui, and Mingmen lasting 30 min was performed once a day for 7 consecutive days, with a depth of 2 mm, intensity of 12-15 mV, frequency of 2 Hz, in SCI+ EA group. The mechanical paw withdraw threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before developing the model and 3, 7, 14, 21 and 28 days after developing the model, and the motor function was assessed using the Basso-Beattie-Bresnahan (BBB) score. The recovery of motor function was assessed using footprint analysis at 28 days after developing the model. After the final behavioral testing, the rats were sacrificed, and spinal cord tissues were harvested to observe the pathological changes of the spinal cord tissues using hematoxylin-eosin staining, to detect the expression of P2X4R and phosphorylated NF-κB p65 (p-NF-κB p65) (by immunohistochemical analysis and Western blot) and to determine contents of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay). Results:Compared with the baseline measured at 1 day before developing the model, the MWT and BBB scores were significantly decreased and the TWL was shortened at each time point after developing the model in SCI group and SCI+ EA group ( P<0.05). Compared with S group, the MWT and BBB scores were significantly decreased and the TWL was shortened at each time point after developing the model, the expression of P2X4R and p-NF-κB p65 in spinal cord tissues was up-regulated, and the contents of TNF-α, IL-1β and IL-6 were increased in SCI group ( P<0.05). Compared with SCI group, the MWT and BBB scores were significantly increased and the TWL was prolonged at 14, 21 and 28 days after developing the model, the expression of P2X4R and p-NF-κB p65 in spinal cord tissues was down-regulated, and the contents of TNF-α, IL-1β and IL-6 were decreased ( P<0.05), and the pathological damage of spinal cord tissues was alleviated and footprints were reduced in SCI+ EA group. Conclusions:The mechanism by which EA alleviates SCI may be related to the inhibition of the activation of the P2X4R/NF-κB signaling pathway and the reduction in the inflammatory response in rats.

Objective To evaluate the practical application of the Media Fill Test(MFT)in assessing aseptic compounding competency of personnel in Pharmacy Intravenous Admixture Services(PIVAS).Methods A multicenter controlled study was conducted across six tertiary hospitals(center ①-⑥)in China.Participants were divided into an inexperienced group(Group A,n=118)and an experienced group(Group B,n=118),each performing five MFT operations.Positive controls validated medium efficacy.Contamination rates and pass rates were analyzed using chi-square and Fisher's exact tests.Results Valid samples included 584 for Group A and 588 for Group B.The sample pass rate was 66.78%(390/584)in Group A and 91.67%(539/588)in Group B,while personnel pass rates were 46.15%(54/117)and 80.51%(95/118),respectively,with significant intergroup differences(both P<0.01).All centers except Center ⑥ showed significantly higher pass rates in Group B(all P<0.05).Conclusion MFT effectively differentiates technical proficiency levels and is suitable for training evaluation of novice PIVAS staff.

BACKGROUND:Adolescent lumbar disc herniation is the main cause of low back pain in adolescents. At present,most of them are treated by conservativetreatment. When long-term non-surgical treatment attempts,surgery may be necessary to prevent further injury when the patient's symptoms are notsufficiently relieved or when the patient has symptoms of single nerve paralysis or compression of the cauda equina,it is very important to choose a suitable interbody fusion device for the surgical treatment of the patients.OBJECTIVE:To explore the effects of minimally invasive interbody fusion with different heights on lumbar biomechanics in patients with adolescent lumbar disc herniation.METHODS:CT scans of a 17-year-old male patient with adolescent lumbar disc herniation (L4-5 segment herniation) were collected. After the three-dimensional reconstruction of MIMICS,the interbody fusion device equal to and 3 mm higher than the intervertebral space was selected for analysis,so two expandablemixed material interbody fusion devices were designed and reconstructed. Fusion device L:11 mm high front,9 mm high posterior,9 mm wide,28 mmlong,and fusion device H:14 mm high front,11 mm high posterior,11 mm wide,28 mm long and the lumbar fusion device was modeled. The fusion deviceand lumbar spine model were optimized,inversely modeled,and then imported into ABAQUS,and finally the 3D model of lumbar fusion was obtained.The physiological activities of the human body were simulated,such as lumbar extension,forward bending,right bending,and left bending,to obtain thecorresponding stress contours. The biomechanical characteristics of the L4-5 vertebra under seven different working conditions were observed.RESULTS AND CONCLUSION:(1) The maximum stress of the two kinds of fuses was in the condition of forward bending and backward extension,the stress value of H fuses was (18.27±3.80) Mpa and (15.02±3.24) Mpa;the stress value of L fuses was (9.16±0.05) Mpa and (9.17±1.83) Mpa. The stress values of the end plate of the H-fusion in the extension station were (19.11±4.03) Mpa and (16.32±3.72) Mpa respectively. The stress values of the L-fusion end plate were (9.13±0.01) Mpa and (4.92±1.01) Mpa respectively. (2) The stress of H-type fusing end plate was higher than that of L-type fusing end plate except for L-5 end plate at neutral position (P<0.05). (3) Choosing an interbody fusion device with a height of more than 3 mm in the same intervertebral space has a more stable biomechanics.

Background and Aims:Obesity is often accompanied by symptoms of disordered eating.Although bariatric metabolic surgery can alleviate these symptoms,there are significant individual differences in postoperative outcomes,and effective predictive indicators are lacking.Liver and kidney function,along with lipid profiles,are closely related to metabolic status and may serve as useful markers for preoperative risk stratification and prognosis prediction.This study was conducted to explore the relationship between preoperative metabolic indicators and symptoms of disordered eating,thereby identifying postoperative recovery patterns among obese patients to support individualized management strategies.Methods:A total of 41 obese patients who underwent sleeve gastrectomy at Shanghai Sixth People's Hospital between September 2020 and June 2023 were enrolled,along with 36 healthy volunteers recruited during the same period.Participants completed the Eating Disorder Inventory-2(EDI-2)questionnaire,and serum samples were collected to assess liver function,kidney function,and lipid levels prior to surgery.The Mantel test was used to analyze correlations between metabolic indicators and EDI-2 scores.Latent profile analysis(LPA)was conducted using the indicators significantly correlated with EDI-2 scores to identify subgroups within the obese cohort.Linear mixed models were then applied to examine the trajectories of postoperative symptom changes across subgroups.Results:Levels of cystatin C,cholinesterase,gamma-glutamyl transferase,triglycerides,and apolipoprotein E were significantly higher in the obese group compared to the healthy group(all P＜0.05),and EDI-2 total score was also significantly elevated(P＜0.05);the prealbumin level in the healthy group was significantly higher than that in the obese group(P＜0.05).These six indicators were positively correlated with EDI-2 score(all r＞0.20,P＜0.05).Based on these markers,the LPA classified the obese group into two subgroups,with subgroup 2 exhibiting higher levels of most metabolic indicators than subgroup 1.During the 18-month postoperative follow-up,both subgroups showed reductions in EDI-2 score,but symptom improvement in subgroup 2 occurred later(month 6)compared to subgroup 1(month 4).Conclusion:Preoperative levels of cholinesterase,gamma-glutamyl transferase,prealbumin,triglycerides,and apolipoprotein E may serve as predictive indicators for improvement in disordered eating symptoms.Recovery patterns after bariatric surgery vary among obese patients with different metabolic profiles,highlighting the need for tailored intervention strategies.

BACKGROUND:Glucose metabolism plays a crucial role in maintaining the normal physiological function of the body.Glucose metabolism disorder can lead to a range of health problems.At present,the molecular mechanism of glucose metabolism and potential gene targets in osteoarthritis need to be further studied.OBJECTIVE:To analyze the genes related to glucose metabolism in osteoarthritis by bioinformatics methods,and to verify them by cell experiments in vitro,so as to provide new ideas for prevention and treatment of osteoarthritis from the perspective of glucose metabolism.METHODS:Differentially expressed genes and glucose metabolism related genes were screened out from GEO database and GeneCards database.The genes related to both osteoarthritis and glucose metabolism were obtained.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were used to screen the functions and pathways of these genes.To further investigate the interactions between these genes,a protein-protein interaction network was constructed and computational methods using Cytoscape software were utilized to identify key genes(Hub genes)for osteoarthritis glucose metabolism.In addition,CIBERSORT algorithm was used to analyze immune cell infiltration in GSE98918 data set.Finally,the expression of Hub gene was verified by cell experiment in vitro.RESULTS AND CONCLUSION:A total of 134 osteoarthritis glucose metabolism-related genes were obtained.GO enrichment analysis showed that GO was mainly involved in the reaction of toxic substances,the positive regulation of inflammatory reaction,the reaction of lipopolysaccharide and so on.KEGG enrichment analysis showed that it was closely related to PI3K-Akt signaling pathway,interleukin-17 signaling pathway,and AGE-RAGE signaling pathway in diabetic complications.Macrophages,monocytes,resting natural killer cells,regulatory T cells,and CD8+T cells were the main infiltrating cells obtained by immune infiltration analysis.In vitro cell experiments showed that the expression of Hub genes SERPINF1,TAC1,GLUL,APOE,and TMEM176A in the experimental group was significantly different from that in the control group.The mRNA expression of HLA-DRA was not statistically significant.The results show that SERPINF1,TAC1,Glul,APOE,and TMEM176A may be the key genes of glucose metabolism in osteoarthritis,and may be potential new targets for the prevention and treatment of osteoarthritis.