Objective:To investigate the effect and mechanism of Fuzheng Jiedu Tongluofang on the expression of MMP-2,MMP-9 and invasion of human hepatoma HepG2 cell.Methods:Hepatioma HepG2 cell was treated with Fuzheng Jiedu Tongluofang.The cell viability was measured by CCK8.The invasion of HepG2 was detected by Transwell assay.The expression of MMP-2 and MMP-9 was analyzed by ELISA.Results:In the CCK8 assay,Fuzheng Jiedu Tongluofang could inhibit the proliferation of HepG2 cells at a density and time measure.In the Transwell assay,the inhibitory rate of invasion was 52.45% when treated with Fuzheng Jiedu Tongluofang at a concentration of 2.65 mg/ml. The ELISA assay indicated that the expression of MMP-2 and MMP-9 decreased significantly after treated with Fuzheng Jiedu Tongluofang(P<0.05). Conclusion:Fuzheng Jiedu Tongluofang suppressed the invasion of HepG2 with the possible mechanism of down-regulating the expression of MMP-2 and MMP-9 in HepG2.

Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P＜0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.

Parkinson's disease(PD)is a common neurodegenerative disease mainly affecting old persons.The pathogenesis of PD is still unclear, although it has been studied for many years.Recently, more and more evidences show that the dysfunction of autophagy plays a pivotal role in the pathogenesis of PD.Substantial progress in the genetic research of PD has showed that several pathogenic genes were identified correlated with autophagy, in particular, playing an important role in sporadic cases of PD.Here, we will discuss pathogenic genes-correlated dysfunctional autophagy-lysosomes system, so as to specify the pathogenesis of PD and provide a clue for its treatment.

Objective:To investigate the clinical characteristics of idiopathic hypereosinophilic syndrome (IHES) with gastrointestinal manifestations,and to improve the level of diagnosis and treatment of IHES. Methods:The clinical materials, process of diagnosis and treatment and prognosis of 9 patients diagnosed as IHES with gastrointestinal manifestations were retrospectively analyzed. Results:The average age of 9 patients was (22.66± 12.86)years old,and the ratio of male and female was about 1.25∶ 1. The main clinical manifestations included abdominal pain,diarrhea and abdominal distension.The eosinophil percentages in peripheral blood and bone marrow of the patients were (42.66 ± 19.88 )% and (39.33 + 15.99 )%, respectively.The ascites exudate cytology examination showed eosinophil infiltrated.The results of gastroscope or colonoscope showed mucosal hyperemia and edema,scattered bleeding spots, and dark red granular hyperplasia; the colon was affected frecuently.The histological biopsy confirmed that the mucosal was infiltrated by eosinophils.The abdominal CT of 6 patients showed that the walls of stomach or bowel were thickened.The abdominal symptoms disappeared,and the ascites was absorpted in 9 patients after the treatment of glucocorticoid.After 2 years of follow up,2 patients had relapse, others had no recurrence.Conclusion:Performing the routine diagnosis and treatment of gastrointestinal diseases, the clinicians should consider the possibility of IHES in order to avoid the misdiagnosis and delayed treatment. When IHES is diagnosed,steroid treatment should be performed in preference.

Chronic diseases of the digestive system are becoming more prevalent in the elderly population.To reduce the occurrence of malignant lesions in the upper gastrointestinal tract, it is important to focus on diagnosing and monitoring esophageal and gastric mucosal lesions in the elderly.The advancement of gastrointestinal endoscopy technology has led to the development of various endoscopic equipments and techniques.One such technology is confocal laser microendoscopy, which allows real-time biopsy and cytology of the mucosal surface.This technology holds significant value in the real-time diagnosis of conditions like gastroesophageal reflux disease(GERD), Barrett's esophagus, esophageal high-level intraepithelial neoplasia, early esophageal squamous carcinoma, atrophic gastritis, and pre-cancerous gastric mucosa in the elderly.Understanding the application of confocal laser microendoscopy in diagnosing esophageal and gastric mucosal lesions in the elderly is crucial for clinicians in making diagnostic and therapeutic decisions.

Objective To investigate the effect of Echinococcus granulosus cyst fluid(EgCF) on the cytoskeletal rearrangement and phagocytosis and the migration of macrophages induced by lipopolysaccharide(LPS). Methods Peritoneal macrophages of C57BL/6 mice were isolated and cultured in vitro, and divided into control group and LPS group and LPS combined with EgCF group. After 48 hours of treatment, filamentous actin (F-actin) changes were observed with rhodamine-labelled phalloidin staining and fluorescence microscopy; Transwell^TM chamber was used to test cell migration ability and flow cytometry to test cell phagocytosis. After 1 hour of treatment, PI3K and AKT, phosphorylated AKT (p-AKT), Rac1, guanosine triphospho-Rac1 (GTP-Rac1), WASP and Arp2 protein expressions were detected with Western blot analysis. Results Compared with the control group, after LPS stimulation, macrophages were deformed significantly; pseudopodia increased; actin cytoskeleton increased and was more distributed in pseudopodia; the ability of migration and phagocytosis were significantly improved, and the expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 proteins significantly increased. EgCF treatment caused cell shrinkage and disappearance of pseudopodia protrusions of LPS-activated cells, and led to the reduced phagocytic and migratory of cells; the protein expression of PI3K, p-AKT, GTP-Rac1, WASP and Arp2 decreased significantly compared with the LPS group. Conclusion LPS induces the migration and enhances phagocytosis of macrophages while EgCF inhibits these effects, which is related to actin cytoskeleton rearrangement.
Mice ; Animals ; Lipopolysaccharides/pharmacology* ; Echinococcus granulosus/metabolism* ; Proto-Oncogene Proteins c-akt ; Cyst Fluid/metabolism* ; Mice, Inbred C57BL ; Macrophages/metabolism* ; Phagocytosis ; Actins/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Guanosine Triphosphate/pharmacology*

Epithelial-mesenchymal transition (EMT) is involved in both physiological and pathological processes. EMT plays an essential role in the invasion, migration and metastasis of tumours. Autophagy has been shown to regulate EMT in a variety of cancers but not in head and neck squamous cell carcinoma (HNSCC). Herein, we investigated whether autophagy also regulates EMT in HNSCC. Analyses of clinical data from three public databases revealed that higher expression of fibronectin-1 (FN1) correlated with poorer prognosis and higher tumour pathological grade in HNSCC. Data from SCC-25 cells demonstrated that rapamycin and Earle's balanced salt solution (EBSS) promoted autophagy, leading to increased FN1 degradation, while 3-methyladenine (3-MA), bafilomycin A1 (Baf A1) and chloroquine (CQ) inhibited autophagy, leading to decreased FN1 degradation. On the other hand, autophagic flux was blocked in BECN1 mutant HNSCC Cal-27 cells, and rapamycin did not promote autophagy in Cal-27 cells; also in addition, FN1 degradation was inhibited. Further, we identified FN1 degradation through the lysosome-dependent degradation pathway using the proteasome inhibitor MG132. Data from immunoprecipitation assays also showed that p62/SQSTM1 participated as an autophagy adapter in the autophagy-lysosome pathway of FN1 degradation. Finally, data from immunoprecipitation assays demonstrated that the interaction between p62 and FN1 was abolished in p62 mutant MCF-7 and A2780 cell lines. These results indicate that autophagy significantly promotes the degradation of FN1. Collectively, our findings clearly suggest that FN1, as a marker of EMT, has adverse effects on HNSCC and elucidate the autophagy-lysosome degradation mechanism of FN1.
Autophagy ; Cell Line, Tumor ; Female ; Fibronectins ; Humans ; Lysosomes/metabolism* ; Ovarian Neoplasms ; Sequestosome-1 Protein/metabolism* ; Squamous Cell Carcinoma of Head and Neck