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 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.

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.

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:Bone marrow mesenchymal stem cels are crucial for bone and cartilage development and regeneration at a celular level. Insufficient quantity and functional impairment of bone marrow mesenchymal stem cels is widely considered to be one of osteoarthritis causes. OBJECTIVE: To explore the relationship between the functional status of bone marrow mesenchymal stem cels and disease progression in osteoarthritis patients.METHODS: Thirty patients with osteoarthritis were enroled from July 2013 to October 2014, and divided into control, mild osteoarthritis, and severe osteoarthritis groups, with 10 cases in each group. 5 mL bone marrow from the femur or tibia was extracted from each patient to isolate and culture bone marrow mesenchymal stem cels. Proliferation ability of cels at passage 3 was detected using cel counting kit-8; toluidine blue staining was performed at 14 days after chondrogenic induction; real-time PCR was used to detect the mRNA expression of Aggrecan and Col2A1 in the control group after chondrogenic induction. RESULTS AND CONCLUSION:Afterin vitro culture, bone marrow mesenchymal stem cels grew adherently in polygonal and fusiform shape with multiple processes at uniform size. The cytoplasm contained larger particles and the nuclei were ovoid. Most of cels were in cel division phase. The proliferation ability was strongest in the control group and weakest in the severe osteoarthritis group. Cels from the three groups were al at plateau phase after 1 week culture. At 14 days after chondrogenic induction, the cels were polygonal and quasi-circular, and purple metachromatic granules distributed outside of the cytoplasm. The expression of Aggrecan and Col2A1 in the control group displayed an overexpression trend. These findings indicate that the functional status of bone marrow mesenchymal stem cels from osteoarthritis patients is negatively correlated with the severity of disease, which can influence the disease progression in osteoarthritis patients.

BACKGROUND:Can adipose-derived mesenchymal stem cells be used to treat sensorineural deafness caused by hair celldegeneration and loss? OBJECTIVE:To investigate the effect of adipose-derived mesenchymal stem cells from guinea pigs transplanted via the scala tympani on the recovery from sensorineural hearing in an animal model. METHODS:Intraperitoneal injection of gentamicin was performed to prepare sensorineural deafness models in guinea pigs. Then, adipose-derived mesenchymal stem cells from guinea pig were transplanted via the scala tympani. After 1 and 3 weeks, auditory brainstem responses were detected to observe the hearing variation after adipose-derived mesenchymal stem celltransplantation. Meanwhile, the migration and distribution of EDU-labeled adipose-derived mesenchymal stem cells in the cochlea were traced. RESULTS AND CONCLUSION:After 1 and 3 weeks of implantation, the results from auditory brainstem response showed that the hearing was gradual y improved. At 1 week of implantation, most of the cells were distributed in the perilymph and some cells migrated to the Corti organ and attached to the basement membrane. At 3 weeks of implantation, cells migrated and attached to the basement membrane of Corti organ, furthermore, some cells migrated to the cochlear nerve. The longer the time of implantation, the less cellsurvived. The results indicate that adipose-derived mesenchymal stem cells from guinea pig that are transplanted via the scala tympani can directly migrate and survive ultimately to improve the hearing.