Objective　Exploring the role of Nestin in liver repair in mice infected with Echinococcus multilocularis. Methods The expression of Nestin in the liver of normal Nestin-cre/Rosa26-tdTomato transgenic mice at 1, 2 and 8 weeks of birth was detected by immunofluorescence method, and the transgenic mice model of Echinococcus multilocularis infection was established. HE was used to detect the pathological damage of Echinococcus multilocularis infected liver, and the expression of Nestin in the liver after Echinococcus multilocularis infection was detected by immunofluorescence. Transwell experiment detected the migration ability of Nestin positive cells stimulated by EmP. Immunofluorescence assay was used to detect the co localization of Nestin and ALB in the liver of transgenic mice infected with Echinococcus multilocularis, and immunohistochemistry was used to detect the expression of Ki67 in the liver of infected mice. Results A Nestin-cre/Rosa26-tdTomato transgenic mouse genotype was established. Immunofluorescence detection revealed that Nestin expression was highest in the liver of mice at 1 week of birth, and gradually decreased after 2 and 8 weeks (P<0.001); The Nestin content in the liver of mice infected with Echinococcus multilocularis gradually increased after 1, 3, and 5 months of infection, and the difference was statistically significant (P<0.001). Transwell experiments showed that EmP at a concentration of 5 μg/mL enhanced the migration ability of Nestin positive cells (P<0.001). Immunofluorescence detection revealed co localization of Nestin and ALB in the liver of mice infected with Echinococcus multilocularis. Immunohistochemistry showed that the expression level of Ki67 in the liver of infected mice gradually increased with infection time of 1, 3, and 5 months (P<0.001). Conclusion After infection with Echinococcus multilocularis, Nestin expression is enhanced in the liver, and Nestin-positive cells are recruited to participate in the process of liver injury repair.

Echinococcosis is a parasitic disease caused by the larval stage of Echinococcus tapeworms, posing a severe threat to patients' health. Its diagnostic techniques play a vital role in clinical diagnosis and treatment, surgical planning, and prognostic evaluation. Currently, the diagnostic methods for echinococcosis mainly include etiological, immunological, imaging, and molecular biological diagnostic techniques. This article comprehensively reviews existing diagnostic techniques for echinococcosis, analyzes the advantages and limitations of various methods, and explores their application prospects. With continuous advancements in scientific technology, emerging diagnostic approaches are expected to substantially enhance the efficiency and accuracy of echinococcosis diagnosis. These research findings will provide valuable references for the development of rapid clinical diagnostic detection products at the current stage, potentially improving cure rates, alleviating patients' disease burden, and offering robust support for the prevention, control, and treatment of echinococcosis.

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.

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.

Objective:To investigate the iodine nutritional status of pregnant women in Qingdao and the effect of prevention and treatment of iodine deficiency disorders (IDD), so as to provide a basis for residents to supplement iodine scientifically, and take targeted prevention measures and adjust intervention strategies.Methods:In accordance with the requirements of the "National Iodine Deficiency Disorders Surveillance Program (2016 edition)" and "Shandong Iodine Deficiency Disorders Surveillance Program", the cluster sampling method was adopted to select pregnant women from 10 districts (cities) in Qingdao from 2018 to 2020, to investigate their basic information and thyroid disease history. Meanwhile, household edible salt samples and random urine samples were collected to detect iodine content.Results:A total of 3 000 pregnant women were monitored from 2018 to 2020, the median age was 31 years, and the median gestational age was 18 weeks. There were significant differences in the distribution of age, gestational age, whether senile puerpera, and pregnancy in different years ( H/χ 2 = 29.35, 81.03, 65.62, 77.34, P < 0.001). The median salt iodine of edible salt ( n = 3 000) and iodized salt ( n = 2 700) in pregnant women's homes were 23.02 and 23.70 mg/kg, respectively. The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt were 89.59% (2 419/2 700), 90.00% (2 700/3 000) and 80.63% (2 419/3 000). The comparison of qualified rate of iodized salt, coverage rate of iodized salt and consumption rate of qualified iodized salt among different years was statistically significant (χ 2 = 48.09, 36.62, 61.08, P < 0.001), the coverage rate of iodized salt and the consumption rate of qualified iodized salt showed a downward trend year by year (χ 2trent = 35.54, 29.50, P < 0.001). A total of 3 000 urine samples were collected from pregnant women and the median urinary iodine of pregnant women was 147.85 μg/L. The urinary iodine level in the third trimester was lower than that in the first and second trimesters ( P < 0.001). The urinary iodine level in the non elderly group was higher than that in the elderly group ( Z = - 6.66, P < 0.001). The urinary iodine level in the group without thyroid disease was higher than that in the group with thyroid disease ( Z = - 1.99, P = 0.047). The urinary iodine level in iodized salt group was higher than that in non-iodized salt group ( Z = - 2.42, P = 0.015). Conclusions:The iodine nutrition of pregnant women in Qingdao is generally at an insufficient level, and the risk of iodine deficiency is high, which needs attention. In recent years, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in Qingdao have shown a downward trend, and have failed to meet the requirements of national standards. In the future, we should strengthen the monitoring and health education of IDD in pregnant women.

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.

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.

Objective:To investigate the distribution characteristics and related factors of HBsAb in infants born to women with chronic hepatitis B virus (HBV) infection.Methods:A total of 605 infants born to women with chronic HBV infection who met the requirements for inclusion were selected as the subjects. Information about the mother′s previous HBV infection, biochemical indicators during pregnancy, pregnancy complications, information about delivery, and hepatitis B test result after birth were collected. HBsAg and HBsAb at the age of 1 year were determined, and HBsAg and HBsAb at the age of 7 months were retrospectively collected. The factors influencing HBsAb in infants were analyzed by ordered logistic regression.Results:In 605 infants, the infection rate was about 1%. Among them, 6 infants were positive for HBsAg and HBV DNA at 7 months and 1 year of age. Uninfected infants were divided into groups according to HBsAb titers. The result showed that there were significant differences in prothrombin activity (PTA) ( χ2=11.17, P=0.01), positive rate of HBeAg ( χ2=7.87, P=0.049) and HBsAg positive rate at birth ( χ2=10.52, P=0.02) among different groups. Multivariate ordered Logistic regression analysis showed that HBsAg negative at birth was an independent protective factor for HBsAb at 7 months of age ( OR=1.564, 95% CI 1.092-2.239, P=0.015). Logistic regression analysis of HBsAb at 1 year of age showed maternal gestational diabetes mellitus ( OR=1.578, 95% CI 1.126-2.210, P=0.008), infant enhanced immunization ( OR=81.207, 95% CI 31.202-211.352, P < 0.001) and antibody level at 7 months of age ( OR=42.123, 95% CI 22.824-77.739, P < 0.001) were independently associated with HBsAb at 1 year of age. Conclusions:HBsAg negative in venous blood at birth was an independent protective factor for HBsAb at 7 months of age, and enhanced immunization was an independent protective factor for HBsAb at 1 year of age.

Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens p38α, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for p38α. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with TGF-β1 effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human TGF-β1.
Cloning, Molecular* ; DNA, Complementary ; Echinococcosis ; Echinococcus granulosus* ; Echinococcus* ; Humans ; In Vitro Techniques ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Phosphotransferases ; Protein Kinases* ; Signal Transduction

BACKGROUND:The cultivation of mammary gland stem cel s is of great significance for the development of mammary gland and breast cancer.
OBJECTIVE:To seek an easy method to isolate and culture mammary gland stem cel in vitro, and verify the safety of cel s.
METHODS:Mammary epithelial cel s were isolated from normal tissues surrounding breast cancer, and CD49f-and EPCAM-positive cel s were sorted using flow cytometry fol owed by surface marker analysis and cel colony formation ability analysis. Afterwards, real-time fluorescent quantitative PCR was used to detect C-erbB-2 and Maspin mRNA expression in mammary gland stem cel s, breast cancer tissues and normal tissues surrounding breast cancer.
RESULTS AND CONCLUSION:Human mammary gland stem cel s were successful y cultured and highly expressed CD49f and EPCAM, with the presence of mixed colony, pleural epithelial cel colony, and myoepithelial cel colony. c-erbB-2 was lowly expressed while Maspin highly expressed in mammary gland stem cel s. Our experimental findings indicate that the mammary gland stem cel s derived from normal tissue surrounding breast cancer have biological safety.