BACKGROUND:Most rotator cuff injuries occur in the supraspinatus tendon.Clinical treatment of rotator cuff injuries is very limited due to the lack of blood vessels and the complex anatomical structure of the rotator cuff.The rapid development of tissue engineering technology and stem cell biology has brought new hope for improving the quality of tendon repair.OBJECTIVE:To prepare human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffolds by bio-3D printing technology to observe the effect of this scaffold on repairing rotator cuff injury.METHODS:(1)In vitro cell assay:The gelatin microcarrier was prepared.The tissue engineered stem cells were constructed by inoculating human umbilical cord mesenchymal stem cells on the surface of gelatin microcarrier.Gelatin methacrylate hydrogel printing ink was prepared.Tissue engineered stem cells were re-suspended with gelatin methacrylate hydrogel printing ink and put into the bio-ink container of 3D printer for printing.Human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffold was obtained after 5 minutes of blue light irradiation and curing.The activity of human umbilical cord mesenchymal stem cells in scaffolds was detected by dead/alive staining and CCK-8 assay.(2)In vivo animal experiments:A random block design method was used to randomly assign 24 SD rats to 4 groups with 6 rats in each group.No treatment was given in the normal group.The rotator cuff injury model of supratinatus tendon tear was established in the rotator cuff injury group,the simple scaffold group,and the cellular scaffold group.The gelatin methacrylate scaffold and human umbilical cord mesenchymal stem cell/gelatin methacrylatecomposite scaffold were implanted into the tendon injury after the model was made in the simple scaffold group and the cellular scaffold group,respectively.Four weeks after operation,behavioral tests and histopathological morphology observation of supraspinatus tendon of rotator cuff were performed.RESULTS AND CONCLUSION:(1)In vitro cellular assay:The dead/alive staining showed that gelatin microcarrier could reduce the damage of human umbilical cord mesenchymal stem cells caused by 3D printing process.With the extension of culture time,the survival rate of human umbilical cord mesenchymal stem cells increased in the scaffold.The results of CCK-8 assay showed that with the extension of culture time,the activity of human umbilical cord mesenchymal stem cells in the scaffold did not change significantly.(2)In vivo animal experiments:Behavioral test results showed that compared with rotator cuff injury group and simple scaffold group,cellular scaffold group significantly improved limb motor function.The results of hematoxylin-eosin and Masson staining of rotator cuff supraspinatus tendon showed that compared with rotator cuff injury group and simple scaffold group,the muscle fiber arrangement in the cellular scaffold group was more regular;there was no obvious inflammatory cell infiltration,and the percentage of collagen volume decreased.The results of immunofluorescence staining showed that the expression levels of interleukin 6 and tumor necrosis factor α in the rotator cuff supraspinatus tendon were significantly decreased in the cellular scaffold group compared with the rotator cuff injury group and the simple scaffold group.(3)The results showed that bio-3D-printed cell scaffolds encapsulating human umbilical cord mesenchymal stem cells/gelatin methacrylate could promote tissue repair and regeneration of rotator cuff injuries.

BACKGROUND:Most rotator cuff injuries occur in the supraspinatus tendon.Clinical treatment of rotator cuff injuries is very limited due to the lack of blood vessels and the complex anatomical structure of the rotator cuff.The rapid development of tissue engineering technology and stem cell biology has brought new hope for improving the quality of tendon repair.OBJECTIVE:To prepare human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffolds by bio-3D printing technology to observe the effect of this scaffold on repairing rotator cuff injury.METHODS:(1)In vitro cell assay:The gelatin microcarrier was prepared.The tissue engineered stem cells were constructed by inoculating human umbilical cord mesenchymal stem cells on the surface of gelatin microcarrier.Gelatin methacrylate hydrogel printing ink was prepared.Tissue engineered stem cells were re-suspended with gelatin methacrylate hydrogel printing ink and put into the bio-ink container of 3D printer for printing.Human umbilical cord mesenchymal stem cells/gelatin methacrylate composite scaffold was obtained after 5 minutes of blue light irradiation and curing.The activity of human umbilical cord mesenchymal stem cells in scaffolds was detected by dead/alive staining and CCK-8 assay.(2)In vivo animal experiments:A random block design method was used to randomly assign 24 SD rats to 4 groups with 6 rats in each group.No treatment was given in the normal group.The rotator cuff injury model of supratinatus tendon tear was established in the rotator cuff injury group,the simple scaffold group,and the cellular scaffold group.The gelatin methacrylate scaffold and human umbilical cord mesenchymal stem cell/gelatin methacrylatecomposite scaffold were implanted into the tendon injury after the model was made in the simple scaffold group and the cellular scaffold group,respectively.Four weeks after operation,behavioral tests and histopathological morphology observation of supraspinatus tendon of rotator cuff were performed.RESULTS AND CONCLUSION:(1)In vitro cellular assay:The dead/alive staining showed that gelatin microcarrier could reduce the damage of human umbilical cord mesenchymal stem cells caused by 3D printing process.With the extension of culture time,the survival rate of human umbilical cord mesenchymal stem cells increased in the scaffold.The results of CCK-8 assay showed that with the extension of culture time,the activity of human umbilical cord mesenchymal stem cells in the scaffold did not change significantly.(2)In vivo animal experiments:Behavioral test results showed that compared with rotator cuff injury group and simple scaffold group,cellular scaffold group significantly improved limb motor function.The results of hematoxylin-eosin and Masson staining of rotator cuff supraspinatus tendon showed that compared with rotator cuff injury group and simple scaffold group,the muscle fiber arrangement in the cellular scaffold group was more regular;there was no obvious inflammatory cell infiltration,and the percentage of collagen volume decreased.The results of immunofluorescence staining showed that the expression levels of interleukin 6 and tumor necrosis factor α in the rotator cuff supraspinatus tendon were significantly decreased in the cellular scaffold group compared with the rotator cuff injury group and the simple scaffold group.(3)The results showed that bio-3D-printed cell scaffolds encapsulating human umbilical cord mesenchymal stem cells/gelatin methacrylate could promote tissue repair and regeneration of rotator cuff injuries.

Objective:To analyze the different expression of neutrophil's protein from the patients with active pulmonary tuber-culosis(TB)and healthy controls(HC)from the perspective of proteomics,and to explore the diagnosis of the secreted proteins related to neutrophil activation in serum.Methods:Neutrophil protein in peripheral blood of 15 patients with TB and 15 HC was identified by label free quantitative proteomics,which were further validated in by ELISA and nephelometry.Results:A total of 358 differentially expressed proteins were identified in our study,54 differentially expressed proteins related to neutrophil activation or degranulation were focused,and seven proteins AHSG,AACT,C3,AAT,AAG,HP and A1BG were validated in 40 TB patients and 40 HC,and the results confirmed that the following proteins were highly expressed in the serum of the TB group:AACT,C3,AAT,AAG,HP and A1BG,while AHSG was lowly expressed,and only the validation results of AHSG were consistent with the trend of proteomics results.In addition,we further investigated the expression level of AHSG in the diagnosis of 24 patients with active pulmonary TB,24 patients with lung cancer,24 HC,and 16 patients with latent tuberculosis infection(LTBI),and found the expression of AHSG was down-regu-lated in TB group compared with the other groups.The sensitivity and specificity of AHSG in differentiating TB patients from HC patients,lung cancer patients,LTBI patients and non-TB patients in the test set were 83.33%,79.17%;78.00%,91.67%;75.00%,87.50%;75.00%,85.94%.Conclusion:Proteomic profiles of neutrophils from TB and HC are obtained from a proteomic perspective,and further validation shows that AHSG has a good differential diagnostic ability for HC,lung cancer and LTBI.

Objective:To analyze the different expression of neutrophil's protein from the patients with active pulmonary tuber-culosis(TB)and healthy controls(HC)from the perspective of proteomics,and to explore the diagnosis of the secreted proteins related to neutrophil activation in serum.Methods:Neutrophil protein in peripheral blood of 15 patients with TB and 15 HC was identified by label free quantitative proteomics,which were further validated in by ELISA and nephelometry.Results:A total of 358 differentially expressed proteins were identified in our study,54 differentially expressed proteins related to neutrophil activation or degranulation were focused,and seven proteins AHSG,AACT,C3,AAT,AAG,HP and A1BG were validated in 40 TB patients and 40 HC,and the results confirmed that the following proteins were highly expressed in the serum of the TB group:AACT,C3,AAT,AAG,HP and A1BG,while AHSG was lowly expressed,and only the validation results of AHSG were consistent with the trend of proteomics results.In addition,we further investigated the expression level of AHSG in the diagnosis of 24 patients with active pulmonary TB,24 patients with lung cancer,24 HC,and 16 patients with latent tuberculosis infection(LTBI),and found the expression of AHSG was down-regu-lated in TB group compared with the other groups.The sensitivity and specificity of AHSG in differentiating TB patients from HC patients,lung cancer patients,LTBI patients and non-TB patients in the test set were 83.33%,79.17%;78.00%,91.67%;75.00%,87.50%;75.00%,85.94%.Conclusion:Proteomic profiles of neutrophils from TB and HC are obtained from a proteomic perspective,and further validation shows that AHSG has a good differential diagnostic ability for HC,lung cancer and LTBI.

BACKGROUND:The absence of blood vessels in tendon tissue makes tendon repair challenging.Therefore,improving tendon healing and raising the efficacy of stem cell and other therapeutic cell transplantation after tendon damage have become hotspots for research in both clinical and scientific contexts. OBJECTIVE:The stem cells and gelatin microcarrier scaffold were joined to form tissue engineered stem cells.Human umbilical cord mesenchymal stem cells cultured in gelatin microcarriers were used to investigate the therapeutic impact and mode of action on tendinopathy healing in rats in vitro and In vivo. METHODS:(1)In vitro cell experiments:After seeding human umbilical cord mesenchymal stem cells with three-dimensional gelatin microcarriers,the cell vitality and survival were assessed.Human umbilical cord mesenchymal stem cells conventionally cultured were cultured as controls.(2)In vivo experiment:Adult SD rats were randomly assigned to normal group,tendinopathy group,2D group(tendinopathy+conventional culture of human umbilical cord mesenchymal stem cells),and 3D group(tendinopathy+gelatin microcarrier three-dimensional culture of human umbilical cord mesenchymal stem cells),with 6 rats in each group.Four weeks after therapy,animal behavior tests and histopathologic morphology of the Achilles tendon was examined. RESULTS AND CONCLUSION:(1)In vitro cell experiments:the seeded human umbilical cord mesenchymal stem cells on gelatin microcarriers showed high viability and as time went on,the stem cell proliferation level grew.Compared with the control group,3D stem cell culture preserved cell viability.(2)In vivo experiment:Following a 4-week treatment,the 3D stem cell culture group showed a significant improvement in both functional recovery of the lower limbs and histopathological scores when compared to the tendinopathy group.The 2D stem cell culture group also showed improvement in tendinopathy injury,but its effect is not as much as the 3D stem cell culture group.(3)The outcomes demonstrate that human umbilical cord mesenchymal stem cells cultured with three-dimensional gelatin microcarrier can promote the repair and regeneration of tendon injury tissue,and the repair effect is better than that of conventional human umbilical cord mesenchymal stem cells.

The differences in chemical quality characteristics between Xinjiang Rehmannia glutinosa and R. glutinosa were analyzed to provide a theoretical basis for the introduction and quality control of R. glutinosa. In this study, the high performance liquid chromatography(HPLC) fingerprints of 6 batches of Xinjiang R. glutinosa and 10 batches of R. glutinosa samples were established. The content of iridoid glycosides, phenylethanoid glycosides, monosaccharides, oligosaccharides, and polysaccharides in Xinjiang R. glutinosa and R. glutinosa was determined by high performance liquid chromatography-diode array detection(HPLC-DAD), high performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD), and ultraviolet-visible spectroscopy(UV-Vis). The determination results were analyzed with by chemical pattern recognition and entropy weight TOPSIS method. The results showed that there were 19 common peaks in the HPLC fingerprints of the 16 batches of R. glutinosa, and catalpol, aucubin, rehmannioside D, rehmannioside A, hydroxytyrosol, leonuride, salidroside, cistanoside A, and verbascoside were identified. Hierarchical cluster analysis(HCA) and principal component analysis(PCA) showed that Qinyang R. glutinosa, Mengzhou R. glutinosa, and Xinjiang R. glutinosa were grouped into three different categories, and eight common components causing the chemical quality difference between Xinjiang R. glutinosa and R. glutinosa in Mengzhou and Qinyang of Henan province were screened out by orthogonal partial least squares discriminant analysis(OPLS-DA). The results of content determination showed that there were glucose, sucrose, raffinose, stachyose, polysaccharides, and nine glycosides in Xinjiang R. glutinosa and R. glutinosa samples, and the content of catalpol, rehmannioside A, leonuride, cistanoside A, verbascoside, sucrose, and glucose was significantly different between Xinjiang R. glutinosa and R. glutinosa. The analysis with entropy weight TOPSIS method showed that the comprehensive quality of R. glutinosa in Mengzhou and Qinyang of Henan province was better than that of Xinjiang R. glutinosa. In conclusion, the types of main chemical components of R. glutinosa and Xinjiang R. glutinosa were the same, but their content was different. The chemical quality of R. glutinosa was better than Xinjiang R. glutinosa, and other components in R. glutinosa from two producing areas and their effects need further study.
Rehmannia/classification* ; Drugs, Chinese Herbal/chemistry* ; Chromatography, High Pressure Liquid/methods* ; Quality Control

Pyroptosis is an identified programmed cell death that has been highly linked to endoplasmic reticulum (ER) dynamics. However, the crucial proteins for modulating dynamic ER membrane curvature change that trigger pyroptosis are currently not well understood. In this study, a biotin-labeled chemical probe of potent pyroptosis inducer α-mangostin (α-MG) was synthesized. Through protein microarray analysis, reticulon-4 (RTN4/Nogo), a crucial regulator of ER membrane curvature, was identified as a target of α-MG. We observed that chemically induced proteasome degradation of RTN4 by α-MG through recruiting E3 ligase UBR5 significantly enhances the pyroptosis phenotype in cancer cells. Interestingly, the downregulation of RTN4 expression significantly facilitated a dynamic remodeling of ER membrane curvature through a transition from tubules to sheets, consequently leading to rapid fusion of the ER with the cell plasma membrane. In particular, the ER-to-plasma membrane fusion process is supported by the observed translocation of several crucial ER markers to the "bubble" structures of pyroptotic cells. Furthermore, α-MG-induced RTN4 knockdown leads to pyruvate kinase M2 (PKM2)-dependent conventional caspase-3/gasdermin E (GSDME) cleavages for pyroptosis progression. In vivo, we observed that chemical or genetic RTN4 knockdown significantly inhibited cancer cells growth, which further exhibited an antitumor immune response with anti-programmed death-1 (anti-PD-1). In translational research, RTN4 high expression was closely correlated with the tumor metastasis and death of patients. Taken together, RTN4 plays a fundamental role in inducing pyroptosis through the modulation of ER membrane curvature remodeling, thus representing a prospective druggable target for anticancer immunotherapy.
Pyroptosis/immunology* ; Humans ; Endoplasmic Reticulum/immunology* ; Animals ; Nogo Proteins/antagonists & inhibitors* ; Mice ; Cell Line, Tumor ; Xanthones/pharmacology* ; Neoplasms/pathology* ; Mice, Nude

Objective: To explore the effects of different types of acute exercise on the working memory of sedentary college students，so as to provide a basis for exercise intervention. Methods: From April 15 to May 30, 2023, a total of 42 sedentary college students were recruited from one university in Beijing. Using a single blind, completely randomized experimental design, participants were randomly assigned to an open skill exercise group, a closed skill exercise group, or a control group, with 14 participants in each group. The open skill exercise group engaged in 30 minutes of badminton, the closed skill exercise group performed 30 minutes of running, and the control group remained seated for 30 minutes. All participants completed a 2-back working memory task and had their electroencephalogram (EEG) data recorded before and after the intervention. Results: The accuracy rates of the open skill exercise group, closed skill exercise group, and control group (0.90±0.06, 0.94±0.05; 0.88±0.05, 0.94±0.05; 0.85±0.10, 0.90±0.06) showed a significant main effect of time ( F=37.14, P <0.01). Reaction times [(923.65±145.08, 711.56± 140.93 ; 909.59±180.28, 807.85±169.66; 917.05±166.35, 871.86±186.07)ms] showed both a significant main effect of time and a significant interaction between group and time ( F=70.55, 11.83, P <0.01). Repeated measures ANOVA revealed that all three groups improved in accuracy and reaction time compared to pre test values, with no significant difference in accuracy between groups. However, the reaction time of the open skill exercise group was significantly faster than that of the control group ( P <0.05), while there was no significant difference between the closed skill exercise group and the control group ( P >0.05). For EEG data, the P2 amplitude showed a significant main effect of time and a significant interaction between groups and time ( F=10.60, 7.66, P < 0.01 ), with the open skill exercise group exhibiting a higher P2 amplitude than the control group ( P <0.05), while the closed skill exercise group showed no significant difference compared to the control group ( P >0.05). The N2 amplitude showed a significant main effect of time ( F=5.94, P <0.05). The P3 amplitude showed significant main effects of time and electrode position, as well as a significant interaction between groups and time ( F=23.16, 4.53, 5.85, P <0.05), with both exercise groups exhibiting higher P3 amplitudes than the control group ( P <0.05), but no significant difference between the two exercise groups ( P >0.05). Conclusion Open skill exercise is more effective than closed skill exercise in improving the working memory of sedentary college students.

Objective:To discuss the differential expression of microRNA(miR)-125a-5p in peripheral blood mononuclear cells(PBMCs)of the patients with mycobacterium tuberculosis(MTB)infection and its effect on macrophage polarization,and to clarify its clinical significance.Methods:A total of 40 patients with active tuberculosis(ATB)(ATB group),35 patients with latent tuberculosis infection(LTBI)(LTBI group),and 40 healthy physical examinees(control group)clinically diagnosed from July 2022 to June 2023 were selected.The fasting blood samples of the subjects in three groups were collected next morning after 12 h of fasting,and then serum was separated.Enzyme-linked immunosorbent assay(ELISA)method was used to detect the levels of inflammatory factors in the serum of the subjects in various groups.Simultaneously,the PBMCs were extracted from the subjects in various groups;flow cytometry was used to detect the expression levels of CD80 and CD206 proteins in the PBMCs of the subjects in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of microRNA(miR)-125a-5p and interleukin-6(IL-6)mRNA in PBMCs of the subjects in various groups.Results:There were no statistically significant differences in the general information of the subjects among three groups(P>0.05).Compared with control group,the erythrocyte sedimentation rate(ESR),percentages of monocytes(MONO),tumor necrosis factor-α(TNF-α)levels,and interleukin-10(IL-10)levels in serum of the patients in ATB group and LTBI group were significantly increased(P<0.05),and the percentages of lymphocytes(LY)were significantly decreased(P<0.05);compared with ATB group,the ESR and level of IL-10 in serum of the patients in LTBI group were significantly decreased(P<0.05),and the percentage of LY was significantly increased(P<0.05);there were statistically significant differences in the counts of white blood cell(WBC)of the subjects among various groups(P>0.05).The flow cytometry results showed that compared with control group,the expression levels of CD80 and CD206 proteins in PBMCs of the patients in ATB group and LTBI group were significantly increased(P<0.05).Compared with ATB group,the expression level of CD206 protein in the PBMCs of the patients in LTBI group was significantly increased(P<0.05),and the expression level of CD80 protein was significantly decreased(P<0.05).The RT-qPCR results showed that compared with control group,the expression levels of miR-125a-5p and IL-6 mRNA in the PBMCs of the patients in ATB group and LTBI group were significantly increased(P<0.05);compared with ATB group,the expression levels of miR-125a-5p and IL-6 mRNA in PBMCs of the patients in LTBI group were significantly increased(P<0.05).The correlation analysis results showed that the miR-125a-5p expression level was positively correlated with the TNF-α level and IL-6 mRNA expression level(r=0.406,P<0.05;r=0.351,P<0.05),and negatively correlated with the IL-10 level(r=-0.368,P<0.05).The area under the receiver operating characteristic(ROC)curve(AUC)value of miR-125a-5p expression level for diagnosing LTBI patients was 0.89(P<0.01),with a sensitivity of 0.85 and a specificity of 0.88.Conclusion:The expression level of miR-125a-5p in PBMCs of the patients in LTBI group is significantly increased,and it can affect the macrophage polarization to M1,promote the inflammatory response process of macrophages and participate in the occurrence and development of pulmonary tuberculosis.

With the increasing demand for dynamic,real-time and rapid qualitative analysis of chemical composition in areas such as emergency response and space exploration,chip-scale mass spectrometers have attracted significant attention.These devices are expected to drive the integration of mass spectrometry with micro/nano-fabrication and intelligent sensing technologies,fostering profound innovation and breakthroughs in analytical chemistry.As an excellent mass analyzer,the ion trap exhibits numerous advantages,and its miniaturization creates favorable conditions for the high-density integration of miniature mass spectrometers.However,the reduction in ion storage capacity may compromise its sensitivity and dynamic range,rendering the study of ion unidirectional ejection in highly miniaturized ion traps of significant practical importance.In this work,a research was conducted on achieving efficient ion unidirectional ejection while maintaining high mass resolution in the extremely miniature hyperbolic linear ion trap(M-HLIT)with a field radius of 1 mm,and an electric field compensation method was proposed,which combined asymmetric electrode stretching and unbalanced RF voltage to achieve high-precision optimization of the electric field composition.Simulations showed that in an ideal structure,this method achieved 100％unidirectional ejection efficiency with the mass resolution of 518,significantly outperforming traditional asymmetric structure method(365)and unbalanced voltage method(321).Following the introduction of ion ejection slots,further optimization through bidirectional stretching and electrical parameters improved the resolution to 790 while maintaining a unidirectional ejection efficiency of 93％.This method eliminated the requirement for additional excitation voltage,offering an ideal solution for the miniature mass analyzer with high detection performance of chip-level mass spectrometers.