Objective To investigate the clinical factors affecting the risk of distant metastasis in patients with gallbladder carcinoma(GBC)and to construct a predictive model for metastasis risk.Methods Retrospective analysis was conducted on 4979 GBC patients from the surveillance,epidemiology,and end results(SEER)database from January 2000 to December 2021,which were divided into a training set(n=3485)and an internal validation set(n=1494)in a 7∶3 ratio.Additionally,47 GBC patients from Hefei First People's Hospital from January 2009 to December 2024 were collected as an external validation set.In the training set,univariate and multivariate analyses were performed to identify independent predictors of distant metastasis in GBC and to construct a predictive model.The predictive ability of the model was assessed by using area under the receiver operating characteristic curve.The model's calibration and clinical utility were evaluated through calibration curves and decision curve analysis.Results The results of univariate and multivariate analyses revealed that Caucasian,tumor size,T stage,and N stage were independent risk factors for distant metastasis in patients with GBC(P＜0.05).The predictive model constructed based on these factors had an AUC of 0.727,indicating good predictive performance.In the training set,internal validation set,and external validation set,the predictive results of this model showed good consistency with the actual situation.Conclusion The nomogram established by using SEER database can accurately predict the distant metastasis status in patients with GBC and demonstrates good clinical applicability.It assists clinicians in intuitively assessing the distant metastasis rate in GBC patients,thereby facilitating the formulation of personalized treatment plans.

Objective To investigate the clinical factors affecting the risk of distant metastasis in patients with gallbladder carcinoma(GBC)and to construct a predictive model for metastasis risk.Methods Retrospective analysis was conducted on 4979 GBC patients from the surveillance,epidemiology,and end results(SEER)database from January 2000 to December 2021,which were divided into a training set(n=3485)and an internal validation set(n=1494)in a 7∶3 ratio.Additionally,47 GBC patients from Hefei First People's Hospital from January 2009 to December 2024 were collected as an external validation set.In the training set,univariate and multivariate analyses were performed to identify independent predictors of distant metastasis in GBC and to construct a predictive model.The predictive ability of the model was assessed by using area under the receiver operating characteristic curve.The model's calibration and clinical utility were evaluated through calibration curves and decision curve analysis.Results The results of univariate and multivariate analyses revealed that Caucasian,tumor size,T stage,and N stage were independent risk factors for distant metastasis in patients with GBC(P＜0.05).The predictive model constructed based on these factors had an AUC of 0.727,indicating good predictive performance.In the training set,internal validation set,and external validation set,the predictive results of this model showed good consistency with the actual situation.Conclusion The nomogram established by using SEER database can accurately predict the distant metastasis status in patients with GBC and demonstrates good clinical applicability.It assists clinicians in intuitively assessing the distant metastasis rate in GBC patients,thereby facilitating the formulation of personalized treatment plans.

Antimicrobial peptides (AMPs) are small molecular peptides widely existing in the innate immunity of organisms, serving as the first line of defense. Natural AMPs possess various biological activities and are difficult to develop drug resistance. However, they are easily broken down by digestive enzymes in the body. In recent years, increasing methods have been reported to enhance the stability of AMPs, including incorporation of unnatural amino acids, chemical modifications, strategic avoidance of enzyme cleavage sites, cyclization, and nano peptide design. This review summarizes the methods for improving the stability of AMPs against protease degradation, aiming to provide references for further research in this field.
Antimicrobial Peptides/pharmacology* ; Humans ; Peptide Hydrolases/metabolism* ; Protein Stability ; Antimicrobial Cationic Peptides/chemistry* ; Anti-Infective Agents/chemistry*

Objective:To study the effects of different single cell phenotypes on the prognosis of patients with intrahepatic cholangiocarcinoma by using spatial analysis, providing clues for obtaining potential immunotherapeutic targets.Methods:This study was a retrospective cohort study. A total of 41 intrahepatic cholangiocarcinoma (ICC) patients who underwent surgery in Beijing Youan Hospital Affiliated to Capital Medical University from February 2013 to June 2019 were enrolled. According to the 5-year survival situation, the patients were divided into survival group ( n=10) and death group ( n=31). A metal label-based tissue imaging mass panel containing 36 related markers was designed and constructed for staining different components in tumor samples. Through the analysis of the type and quantity of different metacluster and spatial location information and combined with the clinical outcomes of patients with information, certain metaclusters were found related to the prognosis of patients. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), paired t-test was used for comparison between groups. Measurement data with skewed distribution were expressed as median, and rank sum test was used for comparison between groups. The chi-square test was used to compare the counting data between groups. Results:36 biomarkers of 41 ICC patients were located and quantified to generate 1 476 single-cell resolution histological images. The expression information of various markers was analyzed by t-distributed Stochastic Neighbor Embedding (tSNE), and subgroups annotations (1-29) were added. It revealed that the density of metacluster 7(CD8 + T cells) was lower in survival group. The density of metacluster 16(Bcl-2 + CK7 + cancer cells) within tumors, as well as the density of metacluster 3(Vista + GB + CD11b + neutrophils) within stroma were higher in death group. Conclusion:The density of metacluster 7(Activated CD8 + T cells), metacluster 16(Bcl-2 + CK7 + tumor cells) and a novel neutrophil metacluster 3(Vista + GB + CD11b + neutrophils) correlated with ICC patients prognosis.

Objective:To investigate the prevalence of work-related musculoskeletal disorders (WMSDs) in rehabilitation therapists.Methods:In September 2023, the computer searched Chinese and English databases such as CNKI database, Wanfang Data Knowledge Service Platform, VIP Chinese Science and Technology Journal Database, Chinese Biomedical Literature Database, PubMed, Cochrane Library, Web of Science, Embase database, etc. The retrieval time was from the establishment of the database to September 2023. Musculoskeletal disorders, rehabilitation therapists were selected as search terms, and the author, publication time, prevalence rate, and study type were analyzed. The Agency for Healthcare Research and Quality (AHRQ) tool and The Newcastle-Ottawa Scale (NOS) were used to evaluate the quality of the literature. Stata 15.0 software was used for meta-analysis.Results:A total of 28 literatures were included, including 8039 rehabilitation therapists. Meta-analysis showed that the overall prevalence rate of WMSDs among rehabilitation therapists was 72% (95% CI: 64%-80%). The top 3 annual prevalence rates were lower back (51%, 95% CI: 38%-64%), neck (42%, 95% CI: 30%-54%) and shoulder (32%, 95% CI: 23%-41%). The top 3 weekly prevalence rates were lower back (47%, 95% CI: 28%-65%), neck (35%, 95% CI: 14%-55%) and back (30%, 95% CI: 17%-44%) . Conclusion:The higher prevalence of WMSDs in rehabilitation therapists, especially in the lower back, neck, shoulder and back, may be related to the prolonged stouching posture and the continuous or repeated exertion of the upper limbs.

Objective:To investigate the prevalence of work-related musculoskeletal disorders (WMSDs) in rehabilitation therapists.Methods:In September 2023, the computer searched Chinese and English databases such as CNKI database, Wanfang Data Knowledge Service Platform, VIP Chinese Science and Technology Journal Database, Chinese Biomedical Literature Database, PubMed, Cochrane Library, Web of Science, Embase database, etc. The retrieval time was from the establishment of the database to September 2023. Musculoskeletal disorders, rehabilitation therapists were selected as search terms, and the author, publication time, prevalence rate, and study type were analyzed. The Agency for Healthcare Research and Quality (AHRQ) tool and The Newcastle-Ottawa Scale (NOS) were used to evaluate the quality of the literature. Stata 15.0 software was used for meta-analysis.Results:A total of 28 literatures were included, including 8039 rehabilitation therapists. Meta-analysis showed that the overall prevalence rate of WMSDs among rehabilitation therapists was 72% (95% CI: 64%-80%). The top 3 annual prevalence rates were lower back (51%, 95% CI: 38%-64%), neck (42%, 95% CI: 30%-54%) and shoulder (32%, 95% CI: 23%-41%). The top 3 weekly prevalence rates were lower back (47%, 95% CI: 28%-65%), neck (35%, 95% CI: 14%-55%) and back (30%, 95% CI: 17%-44%) . Conclusion:The higher prevalence of WMSDs in rehabilitation therapists, especially in the lower back, neck, shoulder and back, may be related to the prolonged stouching posture and the continuous or repeated exertion of the upper limbs.

Objective:To document any effect of a pulsed electromagnetic field (PEMF) on the regulation of neuropeptide Y (NPY) in nucleus pulposus (NP) tissue, NP cell apoptosis and matrix degradation using rats with intervertebral disc degeneration (IDD).Methods:Eighteen Sprague-Dawley rats were randomly divided into a control group, an IDD model group (the model group), and a PEMF group. IDD was induced in both the model and PEMF groups. Right after the modeling, the PEMF group received 14 days of PEMF treatment, while the control group and model group were given no special treatment. Meanwhile, the primary rat nucleus pulposus cells (NPCs) were cultured using Dulbecco′s Modified Eagle Medium at 37℃ and 5% CO 2. When the fusion rate reached 90% after passage, the NPCs were divided into a control group, a TNF-α model group (referred to as model group) and TNF-α + PEMF group (referred to as PEMF group) and treated accordingly. Eight weeks after the modeling, safranin-o/fast green staining was used to assess any pathological morphology changes. The expression of NPY, neuropeptide Y receptor Y2 (NPY2R), bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), collagen type II (Col-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral disc and the cultivated nucleus pulposus cells of the 3 groups were determined. Results:The intervertebral disc cells in the model group were ruptured and folded, with significantly increased polysaccharide and protein components, and significantly increased bone fibers. In the PEMF group the cell boundaries were clearer, with less fibrin fracture and increased cartilage tissue. NPY was expressed in the fibrous annulus and the nucleus pulposus of the intervertebral disc in the model group. The average expression levels of NPY and NPY2R were significantly higher than in the control group and the model group. Compared with the control group, there was a significant increase in the level of Bax and a significant decrease in the expression of Bcl-2 in the model group, and there was a significant decrease in the level of Bax in the PEMF group. Compared with the control group, there was a significant decrease in the Col-II level but a significant increase in the MMP3 protein expression in the model group. The average Col-II mRNA expression was significantly higher in the PEMF group compared with the model group, but the average MMP3 protein expression was significantly less. Those results are consistent with observations in vivo.Conclusion:PEMF may reverse the imbalance of ECM metabolism and delay IDD degeneration by up-regulating the expression of NPY and Bcl-2, as well as blocking the Bax/Bcl-2 signaling pathway to inhibit apoptosis of nucleus pulposus cells.

Objective:To explore any effect of pulsed electromagnetic field (PEMF) stimulation on intervertebral disc degeneration (IDD).Methods:Forty Sprague-Dawley rats were randomly divided into a control group, an IDD model group, a PEMF group and an observation group. An IDD model was induced in all except those in the control group. Both the PEMF and observation groups were given PEMF stimulation, while the latter was additionally injected with the A2AR agonist CGS-21680. Eight weeks after the modelling any pathological changes in the morphology of the rats′ intervertebral disc tissues were evaluated using saffron solid green staining. The expression of A2AR, cyclic adenosine phosphate (cAMP), protein kinase A (PKA), cysteine aspartate proteolytic enzyme-3 (Caspase-3), type II collagen (COL-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral discs were evaluated.Results:The nucleus pulposus had shrunk, while fibrous tissues and chondrocytes had increased in the IDD model group. In the observation group the nucleus pulposus was intact and of basically normal shape. A2AR mRNA and protein levels were higher in the intervertebral disc tissue of the model group than among the control group, on average, while the levels in the observation group were significantly higher than in the other groups. In the PEMF and observation groups cAMP and PKA mRNA were significantly higher than in the IDD model group. The p38 MAPK and P-P38 MAPK levels of the IDD model group and its average P-P38 MAPK/p38 MAPK ratio were significantly higher than in the control group. In the PEMF and observation groups those indices had decreased to varying degrees, with those of the observation group significantly lower than among the model and PEMF groups on average, except for the p38 MAPK values. Caspase-3 and its mRNA were significantly higher in the model group than in the control group, on average, and those values were significantly lower in the PEMF and observation groups than in the IDD model group. The average MMP3 contents of the IDD model group had increased significantly compared with the control group, while the Col-Ⅱ level had decreased significantly. Compared with the IDD model group, the MMP3 level had decreased but Col-Ⅱ expression had increased in both the PEMF and observation groups, with significant differences between the IDD model and observation groups.Conclusions:The activation of the p38 MAPK signaling pathway by inflammatory factors to induce apoptosis is one of the important reasons for the aggravation of IDD lesions. PEMF combined with A2AR agonists can activate the A2AR/cAMP/PKA signaling pathway, inhibit p38 MAPK phosphorylation, reduce apoptosis of nucleus pulposus cells, and relieve IDD damage.

Objective:To observe any regulatory effect of a pulsed electromagnetic field (PEMF) on A2A adenosine receptors (A2ARs) in the nucleus pulposus of rats with intervertebral disc degeneration (IDD), and to explore any combination with the A2ARs′ agonist-mediating ROS/PI3K/Akt signaling pathway.Methods:Fifty Sprague-Dawley rats were randomly divided into a control group, an intervertebral disc degeneration group (the model group), an A2AR agonist CGS-21680 treatment group (the agonist group), a PEMF group and a PEMF combined with CGS-21680 treatment group (the observation group). IDD was modeled in all except the rats in the control group. 100μL of CGS-21680 (100μg/kg) was injected into the L 5-6 intervertebral discs of the agonist group, while the PEMF group was given 30 minutes of PEMF intervention daily for 14 days at 1.5mT and 75Hz with a pulse width of 150μs. The observation group was injected with CGS-21680 and then given the same PEMF intervention. Primary nucleus pulposus cells from each group (50ng/mL) were cultured and the expressions of 8-OHDG, SOD, MDA and ROS were detected by immunohistochemistry, immunofluorescence or with an ELISA kit. The A2AR, PI3K, AKT and p-AKT protein levels were detected using western blotting. Results:The nucleus pulposus cells and the annulus fibrosus were obviously wrinkled, necrotic and broken in the model group but the annulus fibrosus was intact and the nucleus pulposus was almost normal in the observation group. Compared with the model group, the levels of SOD and A2AR, PI3K, p-AKT and AKT protein were higher in the agonist, PEMF and observation groups, while the expressions of MDA, ROS and 8-OHDG were weaker. The ROS level in the observation group was significantly lower than that in the agonist and PEMF groups, and the phosphorylation level of p-AKT in the observation group was significantly higher than in the agonist and PEMF groups. The average levels of SOD, A2AR, PI3K, p-AKT and AKT protein in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly higher than the IL-1β group′s average, while the average levels of MDA, ROS and 8-OHDG were significantly lower. The ROS levels in the observation group were significantly lower than in the agonist and PEMF groups, while the A2AR protein content and p-AKT phosphorylation in the observation group were significantly greater. The average Bax levels in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly lower than that in the IL-1β group, and the expression of Bcl-2 was significantly increased. There was significantly less apoptosis observed in the observation group than in the agonist and PEMF groups, while the expression of Bcl-2 was significantly higher.Conclusions:PEMF plays an anti-oxidative stress role by up-regulating A2AR activity and reducing ROS generation. Treatment with PEMF and A2AR agonist could further activate the phosphorylation of PI3K/Akt, down-regulate Bax and up-regulate Bcl-2, thus inhibiting the apoptosis of nucleus pulposus cells and alleviating the malignant progression of IDD.

OBJECTIVE To prepare hyperoside mixed nanomicelles （Hyp-F127/TPGS） and optimize its preparation technology，and to investigate its intestinal absorption characteristics. METHODS Hyp-F127/TPGS was prepared by thin film dispersion method. Based on single factor test and Plackett-Burman design ，combined with Box-Behnken response surface method ， the preparation process was optimized and validated using entrapped efficiency （EE）and drug loading （DL）as evaluation indexes ， F127-TPGS mass ratio ，hydration time and the amount of Hyp as factors. The appearance and microscopic morphology of Hyp-F127/TPGS obtained by the optimal technology were observed ，and the particle size ，polydispersity index （PDI）and Zeta potential were also determined. The critical micelle concentration （CMC）of blank micelle （F127/TPGS），in vitro release behavior and preliminary stability of Hyp-F 127/TPGS were investigated ，and absorption characteristics of Hyp-F 127/TPGS were investigated by in situ unidirectional intestinal perfusion model. RESULTS The optimal preparation technology of Hyp-F 127/TPGS included F127-TPGS mass ratio of 2∶1，hydration time of 2 h，and Hyp amount of 9 mg. Results of three validation tests showed that the EE of Hyp-F 127/TPGS was （87.20±0.99）％，and the DL was （5.02±1.20）％，deviations from predicted values were 0.92％ and 2.39％. The micelles prepared by optimal technology were yellow ，clear and transparent solution ，with good Tyndall effect ；under transmission electron microscope ，they were spherical ，complete and evenly distributed ；the particle size was （15.02±0.16）nm， the PDI was 0.092±0.031，and the Zeta potential was （－6.67±1.47）mV. The CMC of F 127/TPGS was 21 μg/mL，Hyp-F127/ TPGS was stable after 4 weeks of storage at 4 ℃，and the cumulative release rates of Hyp-F 127/TPGS and Hyp control were （66.30±2.93）％（96 h）and（99.24±0.27）％（60 h），respectively. Hyp-F 127/TPGS and Hyp reference were absorbed in each intestinal segment ，and the main absorption sites were jejunum and duodenum respectively ；drug absorption rate constant andapparent absorption coefficient of the former were significantly higher than those of the latter （P＜0.05 or P＜0.01）. E-mail：zhangyuhangxz@163.com CONCLUSIONS The optimized preparation technology of Hyp-F127/TPGS is stable and feasible ；prepared Hyp-F 127/ TPGS shows a sustained -release effect ，which promotes the intestinal absorption of H yp to a certain extent.