1.Three-dimensional gelatin microspheres loaded human umbilical cord mesenchymal stem cells for chronic tendinopathy repair
Dijun LI ; Jingwei JIU ; Haifeng LIU ; Lei YAN ; Songyan LI ; Bin WANG
Chinese Journal of Tissue Engineering Research 2025;29(7):1356-1362
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.
2.Separation of chemical constituents of Tibetan medicine Fallopia aubertii L.Henry Holub by two-dimensional liquid chromatography
Shenghui SHI ; Xiao LIU ; Dong CHEN ; Dijun JI ; Qian MA ; Yongchang LU
Journal of Pharmaceutical Practice and Service 2025;43(9):443-448
Objective To study the chemical constituents of Fallopia aubertii L.Henry Holub. Methods The chemical constituents of Fallopia aubertii L.Henry Holub. were separated and purified by online two-dimensional preparative liquid chromatography and identified by physical and chemical constants and spectral analysis. The inhibitory activities on xanthine oxidase were determined by ultraviolet spectrophotometry. Results Ten compounds were isolated from the extract of Fallopia aubertii L.Henry Holub, including isotachioside(1), 3,4,5-trimethoxyphenyl-(6'-O-galloyl)-O-β-D-Glucopyranoside(2), 1-hydroxy-,4,5-1-O-[6'-O-(4''-carboxy-1'',3'',5'trihydrotrimethoxyphenylxy)-phenyl]-β-D-glucopyranoside(3), myricetrin(4), myricetin(5), rutin(6), quercetin-3-O-β-D-galactoside(7), quercetin-3-O-β-D-glucopyranoside(8), lyciumideA(9), and N-trans-Feruloyltyramine(10). The inhibitory activity test results showed that the IC50 of compound 5 was 15.92 μmol/L, and the IC50 of compound 6 was 87.36 μmol/L. Conclusion Compounds 1,2,3,4 and 8 were isolated from Medicago polymorpha for the first time. Compounds 5 and 6 had xanthine oxidase inhibitory activity.
3.Investigation and disposal of a pseudo-outbreak of Pseudomonas fluorescens detected from bronchoalveolar lavage fluid
Feng CHEN ; Dijun LIU ; Renjian HUANG ; Min HOU ; Ziyun HUANG ; Kai-jia WANG ; Xingtao YI
Chinese Journal of Infection Control 2025;24(9):1307-1313
Objective To investigate the causes for the continuous detection of Pseudomonas fluorescens(P.fluo-rescens)from bronchoalveolar lavage fluid(BALF)in pediatric department of a hospital,formulate intervention measures and evaluate its effectiveness,and provide basis for improving the whole process infection control of fiber bronchoscopy.Methods Epidemiological investigation was conducted on three children from whose BALF P.fluorescens were detected in May 3-6,2024.The comprehensive methods were adopted,including case revie-wing,on-site process tracking,environmental hygiene monitoring,laboratory testing on disinfectant sterilization effect,fiber bronchoscope structure maintenance and checking,etc.Risks were identified and targeted interventions were implemented.Results Among the 5 pediatric patients who underwent fiber bronchoscopy within 4 days,P.fluorescens was detected from BALF of 3 cases,with a detection rate of 60.0%.The children were 5-8 years old and were admitted to the hospital due to lobar pneumonia.They underwent fiber bronchoscopy from the day of admission to the second day,and bacterial strains were clinically determined to be contaminated strains.Environ-mental sampling showed that the detection rate of P.fluorescens at sampling points such as fiber bronchoscope and enzyme solution storage tank was 15.7%(8/51).After implementing intervention,no target bacteria were detected again,and the difference was statistically significant(P<0.05).From January 1 to May 2,2024,71 BALF from pediatric department were not detected P.fluorescens;From May 3 to 6,among 5 detected BALF,3 were detected P.fluorescens;After intervention(May 16 to December 31),no specimen was detected P.fluorescens.Conclusion This event is a pseudo-outbreak caused by fiber bronchoscope damage as well as improper cleaning and disinfection procedures.Through collaborative investigation and timely intervention by multiple departments,the event was ef-fectively controlled.
4.Investigation and disposal of a pseudo-outbreak of Pseudomonas fluorescens detected from bronchoalveolar lavage fluid
Feng CHEN ; Dijun LIU ; Renjian HUANG ; Min HOU ; Ziyun HUANG ; Kai-jia WANG ; Xingtao YI
Chinese Journal of Infection Control 2025;24(9):1307-1313
Objective To investigate the causes for the continuous detection of Pseudomonas fluorescens(P.fluo-rescens)from bronchoalveolar lavage fluid(BALF)in pediatric department of a hospital,formulate intervention measures and evaluate its effectiveness,and provide basis for improving the whole process infection control of fiber bronchoscopy.Methods Epidemiological investigation was conducted on three children from whose BALF P.fluorescens were detected in May 3-6,2024.The comprehensive methods were adopted,including case revie-wing,on-site process tracking,environmental hygiene monitoring,laboratory testing on disinfectant sterilization effect,fiber bronchoscope structure maintenance and checking,etc.Risks were identified and targeted interventions were implemented.Results Among the 5 pediatric patients who underwent fiber bronchoscopy within 4 days,P.fluorescens was detected from BALF of 3 cases,with a detection rate of 60.0%.The children were 5-8 years old and were admitted to the hospital due to lobar pneumonia.They underwent fiber bronchoscopy from the day of admission to the second day,and bacterial strains were clinically determined to be contaminated strains.Environ-mental sampling showed that the detection rate of P.fluorescens at sampling points such as fiber bronchoscope and enzyme solution storage tank was 15.7%(8/51).After implementing intervention,no target bacteria were detected again,and the difference was statistically significant(P<0.05).From January 1 to May 2,2024,71 BALF from pediatric department were not detected P.fluorescens;From May 3 to 6,among 5 detected BALF,3 were detected P.fluorescens;After intervention(May 16 to December 31),no specimen was detected P.fluorescens.Conclusion This event is a pseudo-outbreak caused by fiber bronchoscope damage as well as improper cleaning and disinfection procedures.Through collaborative investigation and timely intervention by multiple departments,the event was ef-fectively controlled.
5.Correlation Analysis Between Anti-cerebral Ischemia Oxidative Damage and Contents of Active Components in Characteristic Processed Products with Porcine Cardiac Blood and Other Processed Products of Salviae Miltiorrhizae Radix et Rhizoma from Menghe Medical School
Zhen ZENG ; Yuanpei LIAN ; Jiali CAI ; Chunyan YIN ; Dijun WANG ; Li ZHU ; Chanming LIU ; Wei HUANG ; Xiaojing YAN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):162-171
ObjectiveTo analyze the correlation between 11 small molecule active components and 1 protein component of characteristic processed products with porcine cardiac blood and other products of Salviae Miltiorrhizae Radix et Rhizoma(SMRR) from Menghe medical school and anti-cerebral ischemic oxidative damage, and to identify its key component markers of characteristic processed products with porcine cardiac blood for anti-cerebral ischemic oxidative damage. MethodHigh performance liquid chromatography(HPLC) was established to simultaneously determine the contents of 11 active ingredients in SMRR and its processed products[processed with porcine cardiac blood, porcine blood, wine and transferrin(Tf) in porcine cardiac blood], and the content of Tf in different processed products of SMRR was detected by enzyme-linked immunosorbent assay(ELISA). Furthermore, A zebrafish ischemic stroke model was constructed to evaluate the effects of different processed products of SMRR on the behavioral trajectory of cerebral ischemic zebrafish, the neuronal damage of transgenic zebrafish Tg(elavl3:eGFP) brain, as well as the levels of malondialdehyde(MDA) and superoxide dismutase(SOD) in the brain tissues. The hippocampal neurons oxygen-glucose deprivation/reoxygenation(OGD/R)-induced ischemia-hypoxia model was constructed to evaluate the effects of different processed products of SMRR on oxidative damage of neuronal cells by taking lactate dehydrogenase(LDH), reactive oxygen species(ROS), MDA and SOD as indexes. Finally, principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and Spearman correlation analysis were used to analyze the 11 small molecule active components and 1 protein component with efficacy indicators, in order to screen the key components of the characteristic processed products with porcine cardiac blood for cerebral ischemic oxidative damage. ResultCompared with the raw products, the contents of water-soluble and fat-soluble components in processed products of SMRR increased to different degrees, while the content of salvianolic acid A decreased. Compared with the wine-processed products, the contents of salvianolic acid B, danshensu, rosmarinic acid and other components in the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products were increased, while the content of salvianolic acid A was decreased. ELISA results showed that there was no significant difference in Tf content between the porcine cardiac blood-processed products, porcine blood-processed products, Tf-processed products. Pharmacological results showed that different processed products of SMRR could improve the behavioral deficits, brain neuronal injury and oxidative stress after ischemic stroke in zebrafish, and the effect of the porcine cardiac blood-processed products was most pronounced. PCA results showed that salvianolic acid B, salvianolic acid A, rosmarinic acid, lithospermic acid, danshensu, tanshinone ⅡA, caffeic acid, cryptotanshinone and tanshinone Ⅰ were the main contributing components of SMRR and its processed products. And the results of correlation analysis showed that the contents of cryptotanshinone, rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and tanshinone Ⅰ were negatively correlated with MDA level in zebrafish brain tissue, while the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B and Tf were positively correlated with SOD level, and the contents of rosmarinic acid, caffeic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA, tanshinone Ⅰ, danshensu, Tf were positively correlated with neuronal fluorescence intensity in the zebrafish brain. And the contents of lithospermic acid, protocatechuic aldehyde, rosmarinic acid, dihydrotanshinone Ⅰ, salvianolic acid B, tanshinone ⅡA and Tf were negatively correlated with LDH, ROS and MDA levels and positively correlated with SOD level. ConclusionThere are differences in the anti-ischemic oxidative damage effects of SMRR and its different processed products, among which the porcine cardiac blood-processed products has the strongest effect on improving oxidative damage, which may be related to the content changes of salvianolic acid B, danshensu, rosmarinic acid and other components. This study can provide a basis for clarifying the quality markers of SMRR processed with porcine cardiac blood for cerebral ischemia and elucidating its processing mechanism.
6.Proteomic Research on Characteristic Processing Materials of Porcine Cardiac Blood and Porcine Blood from Menghe Medical School and Their Effect on Anti-microglial Inflammatory Damage of Salviae Miltiorrhizae Radix et Rhizoma
Nan LIU ; Yuanpei LIAN ; Chunyan YIN ; Jiali CAI ; Dijun WANG ; Li ZHU ; Chanming LIU ; Wei HUANG ; Xiaojing YAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(18):137-145
ObjectiveProteomics was used to investigate the protein differences between porcine cardiac blood(PCB) and porcine blood(PB) from Menghe medical school and to compare the effects of both on the microglial inflammation of Salviae Miltiorrhizae Radix et Rhizoma(DS). MethodNanoliquid chromatography-quadrupole orbitrap mass spectrometry(nLC-MS/MS) and bioinformatics were utilized to compare the proteomic differences of PCB and PB in simulated gastrointestinal digestion. Furthermore, Western blot was used to verify the contents of some shared proteins and differential proteins identified in PCB and PB. In addition, BV2 neuroinflammation model constructed by corticosterone(CORT) and lipopolysaccharide(LPS) was applied to detect the intervention effects of PCB and PB on the levels of tumor necrosis factor(TNF)-α and interleukin(IL)-6 in BV2 inflammatory cells of DS. ResultA total of 69 common proteins and 68 differential proteins were identified in PCB and PB, among which the common proteins included transferrin(Tf) with brain-targeting effect, and the differential proteins in the two were 41 and 27, respectively. Western blot validation showed that the difference in the content of the same protein Tf between PCB and PB was not statistically significant, while the difference in the contents of the specific proteins of creatine kinase M and heart-type fatty acid binding protein(H-FABP) were statistically significant(P<0.05). Moreover, in vitro experimental studies revealed that compared with the same concentration of DS group, in addition to the 100 mg·L-1 PB-DS group, PCB-DS and PB-DS groups could significantly inhibit the levels of TNF-α and IL-6 in BV2 inflammatory cells(P<0.05, P<0.01), and PCB-DS group had more significant anti-inflammatory effect than PB-DS group with the same concentration(P<0.05, P<0.01). ConclusionBoth of PCB and PB can enhance the inhibitory effect of DS on the release of inflammatory factors, thus playing a neuroprotective role, and PCB promotes DS inhibition more significantly, which may be due to the existence of the two involved in energy metabolism-related differential proteins, which can lay a foundation for revealing the scientific connotation of the processing of PCB-DS and PB-DS.

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