BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

BACKGROUND:In vitro construction of tissue-engineered intestinal models plays an important role in intestinal regeneration and intestinal disease research.The interaction of intestinal nervous system and intestinal epithelial barrier to maintain body homeostasis is a hot topic in the bionic construction of tissue-engineered intestinal tract.OBJECTIVE:To construct a bionic model that can mimic the enteric nervous system in vivo.METHODS:Using fibroin protein with villus structure as scaffold,human induced neural stem cells solidified with collagen were added to intestinal epithelial cells(Caco-2 and HT29-MTX-E12)for 3-day culture to construct a co-culture system of intestinal epithelial cells and nerve cells(co-culture group).Human induced neural stem cells or intestinal epithelial cells cultured alone that were inoculated with fibroin scaffolds were set as controls.Cell morphology was observed by scanning electron microscopy and hematoxylin-eosin staining.Cell activity was detected by Live/Dead cell staining.Human induced neural stem cell differentiation was detected by β-microtubulin immunofluorescence staining.Intestinal epithelial histological properties and barrier function were detected by microvillin,sucrase-isomaltase,tight junction protein 1,E-calmodulin,and mucin-2 immunofluorescence staining.The function of mucus secretion from intestinal epithelial cells was detected by Alcian blue staining.Alkaline phosphatase staining was performed to detect differentiation of intestinal epithelial cells,at the same time,sucrase-isomaltase,tight junction protein 1,and alkaline phosphatase mRNAs were detected by RT-qRCR.RESULTS AND CONCLUSION:The neuralized intestinal mucosal co-culture model with villi structure was successfully constructed,and neural stem cells and intestinal epithelial cells on the fibroin scaffold showed good cellular activities.After neuralization,the activity of alkaline phosphatase and sucrase-isomaltase in intestinal epithelial cells was enhanced,while the expression level of tight junction protein 1 was up-regulated.To conclude,the neuralized bionic intestinal epithelial model is beneficial to the maturation of intestinal mucosal epithelial cells and the formation of barrier function.

[This corrects the article DOI: 10.1016/j.apsb.2022.03.002.].

An ideal dermal filler should integrate filling, repair, and anti-aging effects, with immediate tissue augmentation, slow degradation, and progressive stimulation of collagen regeneration. However, commonly used hyaluronic acid (HA) hydrogels, while effective for rapid filling, suffer from limited duration of support, weak cell adhesion, and an inability to promote collagen regeneration. Silk fibroin (SF), a natural protein from silkworm cocoons, is known for its excellent cell adhesion and collagen-stimulating abilities. However, its limited gelation capability restricts its potential application as a standalone injectable hydrogel. Based on a complementary strategy, this study combines the rapid gelling properties of HA with the collagen regenerative properties of SF to create a co-crosslinked HA-SF hydrogel. The composite hydrogel merges HA's rapid filling effect with SF's strong tissue adhesion and collagen-stimulating abilities. The formulation, physicochemical properties, degradation, biocompatibility, and filling effects of the HA-SF hydrogel were systematically investigated. HA-SF hydrogel exhibits excellent mechanical properties and ensures long-term support while maintaining injectability. Interestingly, after intradermal injection in the UVB-induced photoaging model, HA-SF hydrogel not only enhances hydrogel-cell interaction but also continues to stimulate collagen regeneration, especially type III collagen. This dual action achieves the biological effects of repair and anti-aging while maintaining the filling effect. Proteomic analysis confirms that repair and anti-aging effects are enhanced by the regulation of skin fibroblasts and modulation of amino acid and lipid metabolism. This composite hydrogel holds strong promise for clinical applications, offering a safer, long-lasting, and more natural injectable filler that combines filling, repair, and anti-aging into one system.

Objective:To detect the expression of sialylated CD15 (CD15s) in the tumor cells of classical Hodgkin lymphoma using a modified immunohistochemical approach.Methods:From 2009 to 2024, 53 cases of classical Hodgkin lymphoma were collected in the Department of Pathology, Peking University Cancer Hospital, in which 21 cases that were CD15-negative or showed only focal weak positivity were selected. Immunohistochemical staining for CD15 was performed on a Leica automated stainer using three different antibody clones (MMA, Carb3, and IHC527). Tissue sections were digested with sialidase at varying concentrations and incubation times, followed by immunohistochemical staining with the MMA clone. Multiplex immunofluorescence was applied for co-staining of CD15 (MMA) and CD30 (JCM182), and analysis was conducted using APTIME and HALO software.Results:There were 30 male patients and 23 female patients, with an age range of 14 to 73 years and a median age of 32(26,46) years. None of the three CD15 antibody clones significantly improved the CD15 positive rate in the 14 completely negative and 7 weakly positive cases, with no notable differences observed among the clones( P>0.05). After sialidase digestion, tissue morphology remained well-preserved. Optimal CD15 staining was achieved with a 1∶1 diluted sialidase incubated at 37 ℃ for one hour. This treatment significantly enhanced the detection rate of CD15 antigen in Hodgkin Reed-Sternberg cells ( P<0.001). Conclusion:Sialidase digestion effectively unveils sialylated CD15 expression in classical Hodgkin lymphoma, markedly improving its detection in HRS cells.

Objective:To investigate the efficacy and safety of avatrombopag in promoting hematopoietic reconstitution after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Method:A retrospective analysis was conducted on 60 recipients with hematological malignancies who underwent allo-HSCT at the Affiliated Hospital of Xuzhou Medical University from January 2022 to August 2023. Recipients with hepatic or renal insufficiency before conditioning, those who received other thrombopoietic agents after allo-HSCT, those with severe respiratory or circulatory system diseases, and those with a history of thromboembolic events were excluded. Among them, 30 recipients who received avatrombopag within 14 days post-transplantation were assigned to the avatrombopag group, while the remaining 30 recipients who did not receive any thrombopoietic agents served as the control group. Clinical characteristics, hematopoietic stem cell engraftment, bone marrow proliferation, transfusion requirements, transplant-related complications, and laboratory adverse events were compared between the two groups.Result:The median platelet engraftment time in the avatrombopag group was 13 days (range: 9~25 days), and the neutrophil engraftment time was 13 days (range: 11~21 days). In the control group, he platelet engraftment time was 15 days (range: 10~51 days), and neutrophil engraftment time was 14 days (range: 10~30 days). The difference in platelet engraftment time between the two groups was statistically significant ( P=0.039). Bone marrow analysis on day 28 post-transplant showed that the proportion of recipients with active bone marrow hyperplasia was 96.7% in the avatrombopag group and 73.3% in the control group ( P=0.030); the median number of megakaryocytes was 30 vs. 6, respectively ( P<0.001); and the proportion of mature platelet-producing megakaryocytes was 44% vs. 26.3% ( P<0.001). Regarding transfusion requirements, the median platelet transfusion volume within 28 days post-transplantation was 4.5 U (range: 2~16 U) in the avatrombopag group and 6.5 U (range: 3~32 U) in the control group ( P=0.007). The time to achieve platelet transfusion independence was 13 days (range: 8~25 days) in the avatrombopag group and 14 days (range: 10~36 days) in the control group ( P=0.026). The median red blood cell transfusion volume in both groups was 4 U, with no significant difference ( P=0.354). Medication adherence in the avatrombopag group was 100%. There were no statistically significant differences between the two groups in terms of incidence of post-transplant infections (70% vs. 83.3%), bleeding (50% vs. 60%), graft-versus-host disease (GVHD) (30% vs. 40%), or abnormal laboratory tests (86.7% vs. 90%) (all P>0.05). Conclusion:The use of avatrombopag after allo-HSCT in patients with hematologic malignancies can promote bone marrow hematopoiesis and platelet engraftment, reduce platelet transfusion volume, and shorten the duration of platelet transfusion dependence. Avatrombopag is well tolerated, and no serious adverse reactions were observed during treatment.

OBJECTIVE To investigate the acute lung injury effects of pulmonary phospholipids and their phospholipase A2(PLA2)decomposition products-lysophospholipids and fatty acids-on mice.METHODS Mice were randomly assigned to the following groups:① solvent control(PBS)and PLA2;② solvent control and glycerol phospholipid groups:1,2-dioleoyl-sn-glycero-3-phosphoserine(DOPS),1,2-dipalmitoyl-sn-glycero-3-phosphoserine(DPPS),1,2-dioleoyl-sn-glycero-3-phosphoethanol-amine(DOPE),1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine(DPPE),1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC),and 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine(SOPC);③ solvent con-trol and fatty acid groups:palmitic acid(PA),oleic acid;④ solvent control and lysophospholipid groups:1-oleoyl-2-hydroxy-sn-glycero-3-phosphoserine(18∶1 LysoPS),1-stearoyl-sn-glycero-3-phosphoserine(18∶0 LysoPS),1-palmitoyl-sn-glycero-3-phosphoserine(16∶0 LysoPS),1-palmitoyl-sn-glycero-3-phos-phoethanolamine(16∶0 LysoPE),1-palmitoyl-sn-glycero-3-phosphocholine(16∶0 LysoPC);⑤ solvent control,PLA2,DPPC,PA,16∶0 LysoPC,16∶0 LysoPS,and 18∶1 LysoPS.Following anesthesia,mice were administered nebulized PBS in the solvent control group,2.1 ug·kg-1 PLA2 in PBS in the PLA2 group and 2.5 mg·kg-1 of the corresponding substance in PBS in other experimental groups.For group①,survival times were recorded and survival curves were plotted.At 1 h post-treatment,lung tissues from groups ①②③④ were collected,photographed to obtain white light images,and subjected to HE staining to assess histopathological changes and pathological scoring.At 2 h post-treatment,pulmonary blood flow in group ⑤ was assessed using laser speckle contrast imaging,arterial blood gas was analyzed with a blood gas analyzer,and lung function was evaluated using whole-body pleth-ysmography.At 6 hours post-treatment,blood cells from group ⑤ were analyzed using an automated hematology analyzer.RESULTS Compared with the solvent control group,severe pathological changes were observed in lung tissues of the PLA2 group,accompanied by extensive inflammatory infiltration and interstitial thickening,with all mice succumbing within 240 min.In mice treated with glyc-erol phospholipids,alveolar structures remained clear,alveolar walls were intact and continuous,and alveolar spaces were translucent,with only occasional minor inflammatory cell infiltration in the septa.No significant pathological alterations were detected in the fatty acid groups.Minor inflammatory cell infiltration was seen in the 16∶0 LysoPE and 16∶0 LysoPC groups.However,such pathological changes as patchy hemorrhage,alveolar interstitial edema,increased alveolar wall thickness,and elevated neutrophil counts were observed in the 18∶1 LysoPS,18∶0 LysoPS,and 16∶0 LysoPS groups.Pathological scores based on HE staining were significantly increased in the 16∶0 LysoPS and 18∶1 LysoPS groups com-pared with the solvent control.The percentage of the lung tissue injury area was also markedly higher in the 16∶0 LysoPS group.A significant decrease in the mean fluorescence intensity of blood flow was observed in the 16∶0 LysoPS group.Arterial partial pressure of oxygen(pO2)was significantly reduced in the PLA2 group,while arterial partial pressure of carbon dioxide(pCO2)was markedly elevated in the 16∶0 LysoPS and 18∶1 LysoPS groups.Lung function tests revealed that the 16∶0 LysoPS group exhibited significant increases in expiratory time,end-expiratory pressure,and enhanced pause,in contrast to significant decreases in tidal volume,expired volume,and minute volume.The 18∶1 LysoPS group also exhibited a significant decline in minute volume.No significant changes in inflammatory cell concentrations were detected in blood,with the exception of neutrophils in the 16∶0 LysoPS group,which showed a significant but physiologically normal increase.CONCLUSION Pulmonary phospholipids and their PLA2-derived fatty acid metabolites do not induce severe lung injury in mice while the lyso-phospholipid metabolites,particularly lysophosphatidylserine,are found to cause significant lung injury.

Objective To investigate washing patterns in human simulated body fluids(SBF)by combining high doses of vancomycin,manuperenan and single compound with bone cement.The antibacterial effect of five common bacteria in orthopaedic infections were observed.Methods Vancomycin 2g and meropenem 3g were combined or alone mixed with ordinary bone cement 20g in aseptic operation.Six copies of antibiotic bone cement pellets were made,immersed in SBF,replaced with SBF buffer every 48 hours,sampled every 24 hours 2 μl drops in methicillin-resistant staphylococcus aureus(MRSA),petries of staphylococcus aureus,pseudomonas aeruginosa,E.coli,and extended-spectrum beta-lactamase(ESBL)-positive E.coli were observed for 24 hours and measured the size of the bactericidal rings of the washing fluid at each time point.The antibacterial effect of different ratios of antibiotic bone cement on 5 common orthopedic infections were compared.Results Compound antibiotic group had a better bactericidal effect,and the drug continues to be given for about 30 days.Wancomycin alone could not cover E.coli,ESBL positive E.coli,pseudomonas aeruginosa,and melopicillin alone had only a partial mild bactericidal effect on MRSA around 3 to 6 days after use.The effect on staphylococcus aureus and pseudomonas aeruginosa lasted only about 15 days.Conclusion The combined antibiotic bone cement has a more comprehensive,longer duration and better bactericidal effect than the alone antibiotic bone cement.

Objective To observe conventional ultrasound and contrast-enhanced ultrasound(CEUS)manifestations of Ewing sarcoma(ES)in children.Methods Fifteen children with pathologically confirmed ES were retrospectively collected.Conventional ultrasound and CEUS characteristics of lesions were analyzed.Results Among 15 cases,ES of bone(ESB)was found in 7 cases,while extraskeletal ES(EES)was observed in the other 8 cases.Solitary tumor was noticed in 14 cases,with a median maximum diameter of 7.50 cm,while multiple abdominal masses were found in 1 case.The tumors had irregular shapes and poorly defined boundaries,with medium echogenicity in 7 cases,low echogenicity in 6 cases,while in other 2 cases present as cystic-solid lesions.CDFI showed sparse blood flow in 11 cases,abundant or slightly abundant blood flow in 2 and 1 case,respectively,while no obvious blood flow was observed in 1 case.Rapid high enhancement and rapid washout were found in all 7 cases underwent CEUS,while patchy no-enhancement areas were detected in 4 cases.Conclusion Conventional ultrasonic manifestations of ES had certain specificities,which demonstrated a rapid enhancement and rapid washout pattern during CEUS and may be accompanied by necrosis.

Objective To investigate the predictive value of geriatric nutritional risk index(GNRI)combined with CHA2DS2-VASc-60 score for short-term prognosis of elderly multimorbid patients with atrial fibrillation.Methods A total of 220 elderly multimorbid patients with non-valvular atrial fibrillation admitted to our hospital from May 2020 to December 2022 were recruited.Clinical data were collected,and GNRI and CHA2DS2-VASc-60 score were calculated.With 98 as a cut-off value of GNRI,the patients were divided into normal nutritional group(109 cases)and nutritional risk group(GNRI ≤ 98,111 cases).Clinical characteristics were compared between the two groups.Multivariate logistic regression analysis was used to identify the influencing factors for compound events in elderly multimorbid patients with atrial fibrillation.Variance inflation factor was adopted for collinearity diagnosis.ROC curve was plotted to evaluate the prognostic value of GNRI,CHA2DS2-VASc-60 score and their combination.Results Compared with the normal nutritional group,the nutritional risk group had significantly advanced age,higher CHA2DS2-VASc-60 score,and larger proportion of concomitant chronic heart failure(P＜0.05).Age(OR=1.228,95％CI:1.112～1.357),GNRI(OR=0.693,95％CI:0.494～0.997)and CHA2DS2-VASc-60 score(OR=1.488,95％CI:1.008～2.194)were influencing factors for the occurrence of complex events(P＜0.05,P＜0.01).The AUC value of GNRI,CHA2DS2-VASc-60 score,and their combination in predicting complex events at the end of 6 months was 0.665(95％CI:0.539～0.791),0.689(95％CI:0.578～0.801),and 0.749(95％CI:0.653～0.844),respectively(P＜0.05).Conclusion Age,GNRI,and CHA2DS2-VASc-60 score are influencing factors for the occurrence of complex events in elderly multimorbid patients with atrial fibrillation.The combination of GNRI and CHA2DS2-VASc-60 score has predictive value for the short-term prognosis of the patients.