1.Preparation of decellularized extracellular matrix-gelatin methacryloyl composite hydrogels and their effects on hepatocyte proliferation
Jing SHI ; Jin CHU ; Tao SUN ; Jin GAO ; Xiaolong HE ; Ning YANG ; Liang LI ; Xue ZHANG ; Hui LIU ; Guodong LYU ; Renyong LIN ; Xiaojuan BI
International Journal of Biomedical Engineering 2025;48(1):47-55
Objective:To prepare decellularized extracellular matrix (dECM)-gelatin methacryloyl (GelMA) composite hydrogels and to study their effects on hepatocyte proliferation.Methods:Hepatic dECM was prepared by elution, and GelMA hydrogel and 10%, 30% and 50% dECM-GelMA composite hydrogels were prepared by pepsin solubilization. The morphology of normal liver and dECM liver was observed by eyes and scanning electron microscopy using hematoxylin-eosin, Sirius red and periodate-Schiff staining, respectively. The internal structure of the dECM-GelMA composite hydrogels was observed by scanning electron microscopy, and the pore diameter was measured. Liver HL-7702 cells were co-cultured with GelMA hydrogel and 10%, 30% and 50% dECM-GelMA composite hydrogels, and the cell proliferation viability was determined by cell counting kit-8. The expression of proliferating cell nuclear antigen (PCNA), Wnt family protein 5a (Wnt5a), β-catenin, extracellular-regulated protein kinase 1/2 (ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2) were detected by Western blotting. Comparisons were made using independent sample t-test or one-factor analysis of variance. Results:After decellularization, the hepatocyte morphology showed rounded depressions, and the extracellular matrix structure was intact. The GelMA hydrogel and 10%, 30% and 50% dECM-GelMA composite hydrogels showed inernally porous structures. The pore diameter increased from (3.06±1.35) μm in the GelMA hydrogel to (16.01±4.02) μm in the 50% dECM-GelMA composite hydrogel. On the 3rd, 5th and 7th day, the relative cell proliferation was higher in the 50% dECM-GelMA composite hydrogel group than that in the GelMA hydrogel group (1.89±0.04 vs 1.53±0.01, 9.36±0.04 vs 3.89±0.09, 7.15±0.27 vs 4.89±0.15, all P<0.05). The relative expression levels of PCNA, Wnt5a, β-catenin, and p-ERK1/2/ERK1/2 proteins in the 50% dECM-GelMA composite hydrogel group were higher than those in the GelMA hydrogel group (2.14±0.04 vs 1.00±0.03, 2.36±0.09 vs 1.00±0.08, 1.45±0.03 vs 1.00±0.04, 1.43±0.04 vs 1.00±0.01, all P<0.05). Conclusions:A dECM-GelMA composite hydrogel can be prepared, which may promote hepatocyte proliferation by upregulating the phosphorylation of ERK1/2 and activating Wnt/β-catenin signaling pathway.
2.Preparation of heparin-modified gelatin methacryloyl microspheres and their liver-targeted delivery of ADSCs
Xiaotong LI ; Jin CHU ; Hongbin ZHANG ; Bowen SHI ; Xue ZHENG ; Junlong XUE ; Liang LI ; Renyong LIN ; Xiaojuan BI
International Journal of Biomedical Engineering 2025;48(5):425-434
Objective:To prepare heparin (Hep)-modified gelatin methacryloyl (GelMA) microspheres and to investigate their application in liver-targeted delivery of adipose-derived mesenchymal stem cells (ADSCs).Methods:GelMA microspheres were modified with Hep to obtain GelMA-Hep microspheres. The surface morphology of the GelMA-Hep microspheres was observed by scanning electron microscopy. The changes of carbon atoms, nitrogen atoms and sulfur atoms on the surface of the GelMA-Hep microspheres were detected by X-ray photoelectron spectroscopy. The surface chemical group composition of the GelMA-Hep microspheres was analyzed by Fourier transform infrared spectrometer. The swelling properties of the GelMA-Hep microspheres were detected by water absorption swelling experiment. Human liver HL-7702 cells transfected with lentivirus were co-cultured with GelMA, GelMA-dopamine (GelMA-dop) and GelMA-Hep microspheres. The effects of microspheres on cell proliferation activity were evaluated by cell counting kit-8 method and live/dead cell staining experiment. The adhesion of microspheres to cells was observed by confocal microscopy. The GelMA-Hep microspheres loaded with ADSCs were injected into C57BL/6 mice through the tail vein, and its efficiency of liver-targeted delivery of ADSCs was observed by a small animal in vivo imaging system. The data were compared by independent sample t test or one-way analysis of variance. Results:The GelMA-Hep microspheres were prepared by modifying the GelMA microspheres with Hep. Compared with the GelMA microspheres, the size of the GelMA-Hep microspheres did not change significantly, and the surface did not collapse and showed some crystalline particles. The binding energy of sulfur atoms on the surface of the GelMA-Hep microspheres increased from 166 eV to 168 eV. On the surface of the GelMA-Hep microspheres, the characteristic peaks of sulfonic acid and sulfate groups of Hep were detected at 1 490 cm ?1 and from 1 135 cm ?1 to 1 050 cm ?1, respectively. The swelling rate of the GelMA-dop microspheres was uniform, while the swelling rate of the GelMA microspheres and the GelMA-Hep microsphere was quite different, but the final swelling mass of the three microspheres tended to be consistent at 5 min. After 12, 24, 36 and 48 h of culture, the relative proliferation of cells in the GelMA-Hep group (1.61±0.29, 1.78±0.05, 2.27±0.08, 2.26±0.33) were higher than those in the negative control group (1.00±0.00, 1.28±0.06, 1.39±0.02, 1.41±0.04) (all P<0.05). After 36 h of culture, the relative proliferation of cells in the GelMA-Hep group was higher than that in the GelMA-dop group (1.63±0.21), with significant difference ( P<0.05). Live/dead cell staining experiment showed that after 12 h of cell culture in the GelMA-Hep group, only a few microspheres had cell adhesion; at 24 h, the cells were densely distributed on the surface of the microspheres. After 36 h, the number of cells increased further. At 48 h, live cells were distributed throughout the microspheres. Confocal microscopy showed that after 24 h of culture, cells adhered to the surface of the microspheres in the GelMA-Hep group and showed a stretched morphology. The liver of the GelMA-Hep+ADSCs group showed strong fluorescence at 0.5 h, and the fluorescence brightness continued to 48.0 h. The number of ADSCs reaching the liver was more than that of ADSCs group and GelMA+ADSCs group. Conclusions:GelMA-Hep microspheres were successfully prepared, which can improve the efficiency of liver-targeted delivery of ADSCs.
3.Diagnostic and coding errors on the first page of medical records from a tertiary hospital in Shenzhen
Yong TAO ; Ning ZHANG ; Wenjie WANG ; Zejin LIN ; Renyong LIN
Modern Hospital 2024;24(11):1710-1712
Objective To analyze flaws in diagnosis and coding of diseases on the first pages of inpatient medical records and explore solutions to improve the accuracy of record-filling in tertiary hospitals.Methods A retrospective analysis was con-ducted to assess the quality control and logical verification data from the first pages of inpatient records at a tertiary hospital,col-lected from January to December 2023.Results A total of 463 flaws were identified,including 99 flaws in primary diagnostic selection,accounting for 21.37%,148 flaws in primary surgical selection accounting for 32.00%,33 omissions in secondary di-agnoses,accounting for 7.12%,and 121 omissions in surgical procedures accounting for 26.12%.Primary diagnostic errors were primarily attributed to failure to specify etiology,with the highest number of 24 cases(24.24%),followed by overgeneral-ized diagnostic descriptions,with 15 cases(15.15%).Totally,169 coding errors were identified,including 61 logic errors(36.09%),58 coding omissions(34.33%),and 30 uncombined coding cases(17.75%).Conclusion There are significant flaws in the coding on the first pages of medical records.It is imperative to strengthen the training for clinicians and coders and enhance quality control through intelligent automation.These measures are crucial for improving the quality control of the first pa-ges of medical records.
4.The effect of the extracellular vesicle loaded polylactic acid microspheres in promoting hepatocyte proliferation
CHU Jin ; Mukexina Mulati ; GAO Jin ; LI Liang ; ZHANG Xue ; LYU Guodong ; LIN Renyong ; BI Xiaojuan
China Tropical Medicine 2023;23(10):1030-
Abstract: Objective To prepare a microparticle delivery system that regulates the release rate of extracellular vesicles (EVs), and to exert long-term enhancement of liver cell proliferation after only one intervention. Methods EVs was extracted by differential centrifugation. The structure of the EVs was observed by transmission electron microscopy and the membrane marker protein of EVs was detected by Western blotting. EVs-PLA microspheres with "core-shell" structure were prepared by emulsion-solvent evaporation method. Scanning and transmission electron microscopy were used to detect the morphology of EVs-PLA microspheres and EVs. The release test detected the release behavior of EVs in EVs-PLA microspheres. Scanning electron microscopy was used to detect the morphological changes of EVs-PLA microspheres at 8 weeks of release. EVs-PLA microspheres were co-cultured with hepatocytes, and Phalloidin/DAPI staining was used to observe the cell morphology and evaluate the cytotoxicity of the microspheres. CCK8-test was used to evaluate the cell proliferation activity. Western blot analysis was used to detect extracellular vesicles membrane marker protein expression. Results Comparing the ability of hepatocyte proliferation in the group treated with EVs-PLA microspheres and the control group, it was found that EVs-PLA microspheres did not cause cell apoptosis and mutation in cell structure, had biocompatibility and no cytotoxicity. The EVs-PLA microspheres with "core-shell" structure regulated the release behavior of EVs, which can continuously release EVs, exerting a continuous biological role in promoting hepatocyte proliferation after a single intervention. Conclusions The EVs-PLA microspheres can control-release EVs and promote hepatocyte proliferation continuously after a single intervention, providing a reference for further exploration of EVs-loaded delivery systems in promoting liver regeneration.
5.Research progress on the role of macrophages in echinococcosis and the regulation of macrophages by parasite-derived molecules
Chinese Journal of Endemiology 2023;42(3):246-251
As a major member of innate immunity, macrophage can eliminate pathogens through cell phagocytosis, antigen presentation and immune regulation, and play an important role in parasitic infections such as Echinococcus. Echinococcus can regulate the function of host macrophages through a variety of parasite-derived molecules, such as protein and nucleic acid molecules, and realize long-term parasitism in the host. This article focuses on the research progress of the role of macrophages in echinococcosis and the regulation of macrophages by parasite-derived molecules.
6.Study on the characteristics of 18F-flurodeoxyglucose uptake in the microenvironment of hepatic alveolar echinococcosis
Abulizi ABUDUAINI ; Aini ABUDUSALAMU ; Bo RAN ; Tiemin JIANG ; Ruiqing ZHANG ; Qiang GUO ; Xiaohong LI ; Jian WANG ; Chuanshan ZHANG ; Renyong LIN ; Hao WEN ; Yingmei SHAO ; Aji TUERGANAILI
Chinese Journal of Hepatobiliary Surgery 2023;29(3):176-180
Objective:To investigate characteristics of the 18F-flurodeoxyglucose ( 18F-FDG) uptake intensity and ranges in distinct hepatic alveolar echinococcosis lesions. Methods:The clinical data of 39 patients with position emission tomography during Jan 2017 to Dec 2019 in the First Affiliated Hospital of Xinjiang Medical University were enrolled. Among them, there were 17 males and 22 females, aging from 15 to 65 years (median 34 years). Lesions were classified into six groups based on heterogenic scales of calcification and liquefaction: A. non-calcified and non-liquefied ( n=7); B. obvious calcified and non-liquefied ( n=7); C. partial calcified and partial liquefied( n=10); D. obvious calcified and partial liquefied ( n=5); E. partial calcified and subtotal liquefied ( n=5); F. obvious calcified and subtotal liquefied ( n=5). Tumor to background ratio (TBR) and width (W) of lesion infiltrative boundary were measured and calculated. Statistical comparison using Mann-Whitney U test as well as correlation analysis was performed. Results:TBR values [ M( Q1, Q3)] for each group were 4.40(3.66, 7.03), 2.55(1.69, 3.60), 3.73(3.37, 5.21), 2.90(2.75, 3.60), 3.80(3.49, 6.36), 2.49(2.21, 3.97), among which A>B, A>D, A>F, C>B, E>B ( U=3.0, 4.0, 4.5, 11.0, 5.0, all P<0.05); From the perspective of the calcification in each group, it was found that the lighter the calcification was, the greater the TBR value was. W values [ M( Q1, Q3)] for each group were [12.5(10.0, 19.5), 11.2(10.5, 12.5), 12.2(10.9, 13.2), 7.8(7.3, 9.3), 10.0(7.3, 13.4), 7.3(6.8, 7.6)] mm, among which A>D, A>F, B>D, B>F, C>D, C>F (all U=0, all P<0.05); According to the degree of calcification and liquefaction of lesions in each group, the lighter the calcification was, the greater the W value was; The heavier the liquefaction was, the smaller the W value was. A mild strength linear correlation has been observed between the TBR value and W value ( r=0.4136, P<0.05). Conclusions:Less calcification and liquefaction implicated higher 18F-FDG uptake intensity and wider range. Radical resection margins and tissue sampling should be individualized based on different lesion features in surgical treatment.
7.A prospective study on implication value of lesion microenvironment's immune cell infiltration range on liver sampling in hepatic alveolar echinococcosis
Aini ABUDUSALAMU ; Abulizi ABUDUAINI ; Tiemin JIANG ; Bo RAN ; Renyong LIN ; Chuanshan ZHANG ; Aji TUERGANAILI ; Hao WEN ; Yingmei SHAO
Chinese Journal of Endemiology 2023;42(10):781-785
Objective:To investigate the immune cell infiltration width on lesion microenvironment (LME) based on different hepatic alveolar echinococcosis lesion features to underlie referential sampling range for experimental and control tissues aiming at avoiding false negativity in basic researches.Methods:Using prospective research methods, from January 2017 to December 2019, patients with hepatic alveolar echinococcosis who were diagnosed and treated surgically at the First Affiliated Hospital of Xinjiang Medical University and met the multi-site sampling method (MSS method) were investigated. A total of 26 cases were included, aged 34 (15, 65) years old, with a gender ratio of 12/14. Lesions were classified into six groups based on heterogenic scales of calcification and liquefaction: A. non-calcified and non-liquefied; B. obvious calcified and non-liquefied; C. partial calcified and partial liquefied; D. obvious calcified and partial liquefied; E. partial calcified and subtotal liquefied; F. obvious calcified and subtotal liquefied. Liver specimens were acquired with 5 mm interval off the lesion shore in LME area using MSS method. Performed immunohistochemical staining of CD3, CD19, CD68, and Masson staining of fibrous tissue, and after pathological evaluation, the layer with a sharp decrease in immune cell infiltration abundance was determined as the maximum infiltration range (width, W value). The experimental group conservatively estimated the maximum sampling range for the integer value of Q1 of W value. The control group conservatively estimated the minimum sampling range for the integer value after Q3 rounding plus 5 mm of W value. Results were analyzed using Kruskal-Wallis H and Mann-Whitney U tests, referential ranges were concluded. Results:Median W values (interquartile) for each group were 20.0 (12.5, 22.5), 15.0 (10.0, 15.0), 10.0 (10.0, 15.0), 5.0 (5.0, 10.0), 12.5 (6.3, 15.0), and 5.0 (5.0, 10.0) mm, among which A > D, A > F, C > D, and C > F ( P≤0.05); from the perspective of calcification, A > C + E, A > B + D + F ( P < 0.05), while A + B > C + D, A + B > E + F ( P < 0.05) from the perspective of liquefaction. In these groups, the experimental group conservatively estimated the maximum distance for sampling to be 12.0, 10.0, 10.0, 5.0, 6.0, and 5.0 mm, while the control group conservatively estimated the minimum distance for sampling to be 28.0, 20.0, 20.0, 15.0, 20.0, and 15.0 mm. Conclusions:Less calcification and liquefaction implicates wider immune cell infiltration range in those lesions. Tissue sampling should be individualized based on different lesion features in basic research to avoid false negativity.
8.Role of CD155 in hepatocyte apoptosis and liver fibrosis induced by Echinococcus multilocularis infection in mice
Junjie SHI ; Kusuman NUERBAITI ; Ning YANG ; Xiaojuan BI ; Renyong LIN ; Ailian ZHANG
International Journal of Biomedical Engineering 2022;45(5):384-389,394
Objective:To investigate the role of CD155 in hepatocyte apoptosis and liver fibrosis in mice infected with Echinococcus multilocularis. Methods:Thirty-six female C57BL/6 mice were randomly divided into a sham surgery group and a model group, with 18 mice in each group. Mice in the model group were injected with protoscolex via the portal vein to create an animal model of E. multilocularis infection. Mice in the sham surgery group were injected with the same amount of saline. The mice were sacrificed at 1 month, 3 months, and 6 months after modeling, and liver samples were collected. Hepatic pathological changes were observed by hematoxylin-eosin staining. Liver fibrosis was detected by Sirius red staining, and expression of Caspase-3 and CD155 in hepatocytes was detected by immunohistochemical staining. The correlation between CD155 expression in hepatocytes and Caspase-3 and liver fibrosis levels were analyzed by Person. Results:There were obvious lesions in the liver of the model group accompanied by severe liver fibrosis. Compared with the sham surgery group, the expression of CD155 and Caspase-3 in mouse hepatocytes at different stages in the model group was significantly increased, and the differences were statistically significant ( P<0.05). The model group's liver fibrosis level was significantly higher at different stages than the sham surgery group, with statistical significance ( P<0.05). In addition, correlation analysis showed that expression of CD155 in hepatocytes was positively correlated with the expression of Caspase-3 ( r=0.956 8; P<0.001; 95% CI: 0.885 5-0.984 1) and that expression of CD155 in hepatocytes was positively correlated with the degree of liver fibrosis( r=0.853 9; P<0.001; 95% CI: 0.643 7-0.944 3). Conclusions:CD155 expression was significantly up-regulated in mouse hepatocytes infected with E. multilocularis at different stages, which was positively correlated with the degree of hepatocyte apoptosis and liver fibrosis, suggesting that CD155 may be involved in the process of hepatocyte apoptosis and liver fibrosis caused by E. multilocularis infection.
9.Current progress on lesion classification of hepatic alveolar echinococcosis
Aini ABUDUSALAMU ; Bo RAN ; Tiemin JIANG ; Ruiqin ZHANG ; Qiang GUO ; Xiaohong LI ; Jian WANG ; Chuanshan ZHANG ; Renyong LIN ; Yingmei SHAO ; Hao WEN ; Aji TUERGANAILI
Chinese Journal of Hepatobiliary Surgery 2021;27(6):461-466
Human alveolar echinococcosis is a chronic infectious disease caused by Echinococcus multilocularis infection. It predominantly injuries the liver and grows like the malignant tumor. The therapeutic options and prognosis depend on types of human alveolar echinococcosis, clinical stages, biological activity, vascular invasion, pathological characteristics, and patient's immune status. However, despite of multiple classification methods, there are still lacking of comprehensive typing schemes. which leads to inappropriate diagnosis and therapy. This research systematically reviewed the recent studies on human alveolar echinococcosis at home and abroad and analyzed the classifications based on ultrasound, computer tomography, magnetic resonance imaging, positron emission computed tomography, serology and pathology, and some novel technologies and summarized the individual advantage and disadvantage for each classification Relationships and their advantages plus disadvantages have been assessed comprehensively. Meanwhile, the possible reference factors or theoretical basis for optimized future classification are proposed, in order to establish a unified classification system to provide guidance for clinical diagnosis and treatment.
10.Effect evaluation of the defluoridation projects in preventing children's dental fluorosis in 134 Regiment, 8 Division, Xinjiang Production and Construction Corps
Li SUN ; Binbin FANG ; Yumin TANG ; Bolin LI ; Hui WANG ; Renyong LIN ; Shunhua WU
Chinese Journal of Endemiology 2018;37(12):981-984
Objective To investigate the prevalence and trend of dental fluorosis of 7-14 years old children in 134 Regiment,8 Division,Xinjiang Production and Construction Corps,and to evaluate the effectiveness of water improvement and fluoride control measures.Methods From 2010 to 2017,using cross-sectional survey,six water allocation places were selected from 134 Regiment,8 Division,Xinjiang Production and Construction Corps,and the fluoride content was determined.Children of 7-14 years old in 2 central primary schools were investigated,and dental fluorosis was examined.Taking 2017 as the benchmark,children born before water improvement were 11-14 years old,children born after water improvement were 7-10 years old.Water fluoride was detected via the ion-selective electrode method.Diagnosis of dental fluorosis was based on the standard of "Dental Fluorosis Diagnosis" (WS/T 208-2011).The detection rate of dental fluorosis was compared by x2 test,and rank sum test was used to compare the severity of the disease.Results A comprehensive water improvement and fluoride reduction project was completed in 134 Regiment,8 Division,Xinjiang Production and Construction Corps in 2007.The detection rate of dental fluorosis in children born before water improvement was 2.65 times higher than that of children born after water improvement [14.43% (101/700) vs 5.44% (33/607),X2 =28.567,P < 0.01].The dental fluorosis index of children born before water improvement was also higher than that of children born after water improvement (0.33 vs 0.11).According to age standardization (based on 2017),there was a significant difference in the detection rate of dental fluorosis among children in different years (x2 =351.300,P < 0.01).The detection rate of dental fluorosis in children decreased from 35.26% in 2010 to 10.25% in 2017.There was a statistically significant difference in the severity of dental fluorosis in children of different years (H =954.033,P < 0.01).The dental fluorosis index of children decreased from 0.71 in 2010 to 0.23 in 2017,and the disease changed from extremely mild fluorosis epidemic to non-fluorosis epidemic.Conclusion After effective water improvement in 134 Regiment,8 Division,Xinjiang Production and Construction Corps,the prevalence of dental fluorosis in children in the disease affected areas has decreased significantly,the effect of defluoridation project is significant.

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