BACKGROUND:Compared with bone marrow transplantation, peripheral blood stem cel transplantation has its own advantages, including rich resources of stem cel s from the peripheral blood, convenient and easy col ection, without anesthesia, smal trauma, easily accepted, high safety, and easy to restore the patient’s hematopoietic system.
OBJECTIVE:To observe the function and safety of autologous peripheral blood mononuclear cel s in the treatment of patients with decompensated cirrhosis.
METHODS:Four patients with decompensated liver cirrhosis were selected from November 2010 to July 2011 in the First Affiliated Hospital of Dalian Medical University, aged 31-67 (averagely 44 years). Among them, three cases had hepatitis B, and another one had autoimmune liver disease. Peripheral blood stem cel s were col ected after being mobilized by granulocyte colony stimulating factor. Then, autologous peripheral blood stem cel s were transplanted via a hepatic artery catheter.
RESULTS AND CONCLUSION:There were no adverse reactions such as fever, bleeding and nausea after peripheral blood stem cel col ection and hepatic artery transplantation. Symptoms such as fatigue, poor appetite and abdominal distension gradual y improved at 1, 3 and 6 months after transplantation. Liver function and liver fibrosis indexes were improved to some extent after transplantation.

BACKGROUND:Bone marrow mesenchymal stem cells are considered as commonly used seed cells to construct tissue-engineered for repair of bone and cartilage defects. It is of great significance for cytology and tissue engineering experiments to study the common problems existing in the basic operation and how to avoid these problems in a timely manner. OBJECTIVE:To summarize the common problems existing in the process of operation in order to provide reliable methods about separation, culture and identification of bone marrow mesenchymal stem cells for beginners and researchers. These can reduce or avoid some errors and problems during operation. METHODS:Sixteen New Zealand white rabbits were selected as experiment objects, and bone marrow mesenchymal stem cells were separated from rabbits by iliac puncture, purified and augmented by using density gradient centrifugation combined with adherent culture method. Then cellmorphology was observed by inverted phase contrast microscope, growth curve detected by MTT method and cellphenotype identified by flow cytometry. RESULTS AND CONCLUSION:We encountered some problems in the process of separation and culture, when we operated the first five rabbits. After careful y summarizing and analysis of the reasons, the operation was successful y completed on the rest 11 rabbits. Bacteria pol ution and cellaging were not found in the process of cellculture. What is more, the cells at passage 3 appeared with high-expression of CD29, and CD44, but low expression of CD14 and CD34. The cellgrowth curve showed that the proliferation activity of cells at passages 3 and 5 was higher than that at passage 10. Although the technology of separation, culture and identification of bone marrow mesenchymal stem cells is mature, the failure wil be happen if we do not pay attention to the details of operation. By strictly carrying out normal operations, we can get high purity of bone marrow mesenchymal stem cells, which lays a good foundation for celland animal experiments in the future.

Theacrine is one kind of natural purine alkaloids, which mainly exists in an unusual Chinese tea known as Kucha. It shows various biological activities, such as anti-depression, sedative and hypnotic effects and anti-inflammatory and analgesic activi-ties. The study on theacrine dates back to a few decades ago. According to the references published in recent years, the resource, preparation, characterization and pharmacological effects of theacrine were reviewed, and its application prospect was also explored.

ObjectiveTo investigate the mechanism of action of Xiaoyao powder combined with Sijunzi decoction, Artemisia capillaris Thunb. decoction, Longdan Xiegan decoction combined with Xiayuxue decoction, Wupi decoction combined with Sijunzi decoction, and Yiguan decoction in the treatment of hepatocellular carcinoma (HCC) based on network pharmacology and molecular docking. MethodsDatabases including TCMSP, TCMID, BATMAN-TCM, and TCM-MESH were used to screen out effective components and predict their targets, and databases including TTD, Drugbank, Disgenet, Liverome, OncoDB.HCC, and GEO were used to investigate HCC-related targets. The drug and disease targets were mapped to obtain the intersecting targets, and the visualization software Cytoscape 3.7.1 was used to construct the core component-intersecting target network and the protein-protein interaction (PPI) network. The core components and key genes were screened out and a survival analysis was performed in the GEPIA database. The active components and key genes screened out were imported into the DockThor online website for molecular docking. In addition, David database was used to perform gene ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the intersecting targets. ResultsThe analysis showed that 110, 19, 154, 121, and 51 active components, respectively, were obtained for the above five classic prescriptions, and the numbers of drug targets were 7426, 1435, 9544, 6619, and 2427, respectively. Finally 4001 HCC disease targets were screened out. There were 260, 169, 276, 242, and 192 intersecting targets, respectively, between the five prescriptions and the HCC disease targets, and the survival analysis on the GEPIA online website obtained the common hub genes of PIK3CA, SRC, MAPK1, MAPK3 (all P＜0.05) and AKT1 (P＞0.05). Quercetin was the common active component of the five prescriptions, and isobavachin and Kanzonol W were the common active components of Xiaoyao powder combined with Sijunzi decoction, Longdan Xiegan decoction combined with Xiayuxue decoction, and Wupi decoction combined with Sijunzi decoction; the results of molecular docking showed that the above three components had a strong ability of binding to PIK3CA and SRC. GO enrichment analysis showed that these targets were involved in various biological processes including drug response, protein phosphorylation, inflammatory response, and angiogenesis, and KEGG enrichment analysis showed that the common pathways involved were cancer pathway, PI3K-Akt signaling pathway, MAPK signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, hepatitis B pathway, and hepatitis C pathway. ConclusionQuercetin, isoflavone, and Kanzonol W have the potential mechanism of action involving multiple targets and pathways in the treatment of HCC.

Objective: To detect the expression of non-coding RNA snord105b in gastric cancer (GC) tissues, sera and cell lines, and its correlation with clinicopathological characteristics of patients with GC as well as its effect on the proliferation of GC cells. Methods: One hundred and twenty pairs of GC tissues and corresponding para-cancerous tissues from patients, who underwent surgery at Department of Surgery, the Fourth Hospital of Hebei Medical University between 2016 and 2017, were collected for this study. The presurgical sera samples from GC patients (n=50) and peripheral venous blood samples from healthy donors (n=30), as well as five gastric cancer cell lines (SGC-7901, AGS, MGC-803, BGC-823, HGC-27) and gastric mucosa normal epithelial GES-1 cells were also obtained. qPCR assay was adopted to detect the expression of snord105b in GC tissues, sera and cell lines. The correlation between snord105b and patients’clinicopathological features was investigated. MTS assay was adopted to detect the effect of snord105b silence or over-expressionon in vitro proliferation of four GC cells. Results: qPCR assay demonstrated that the expression of snord105b in GC tissues, sera and cell lines were significantly higher than that of para-cancerous tissues, sera from healthy donors and GES-1 cells (all P< 0.05). Expression level of snord105b was obviously associated with age,tumor size, differentiation and TNM stages of patients (all P<0.05). MTS assay demonstrated that knockdown of snord105b could suppress the proliferation of GC cells (P< 0.05), while forced-expression of snord105b could promote the proliferation of GC cells (P< 0.05). Conclusion: non-coding RNA snord105b aberrantly expressed in GC tissues, sera, and cells, and its expression was obviously correlated with patients’age, tumor size, differentiation and TNM stages. Snord105b could significantly promote the proliferation of GC cells, which may be used as a potential clinical biomaker for early diagnosis and prognosis of GC.

Chaperone-mediated autophagy (CMA) is a lysosome-dependent selective degradation pathway implicated in the pathogenesis of cancer and neurodegenerative diseases. However, the mechanisms that regulate CMA are not fully understood. Here, using unbiased drug screening approaches, we discover Metformin, a drug that is commonly the first medication prescribed for type 2 diabetes, can induce CMA. We delineate the mechanism of CMA induction by Metformin to be via activation of TAK1-IKKα/β signaling that leads to phosphorylation of Ser85 of the key mediator of CMA, Hsc70, and its activation. Notably, we find that amyloid-beta precursor protein (APP) is a CMA substrate and that it binds to Hsc70 in an IKKα/β-dependent manner. The inhibition of CMA-mediated degradation of APP enhances its cytotoxicity. Importantly, we find that in the APP/PS1 mouse model of Alzheimer's disease (AD), activation of CMA by Hsc70 overexpression or Metformin potently reduces the accumulated brain Aβ plaque levels and reverses the molecular and behavioral AD phenotypes. Our study elucidates a novel mechanism of CMA regulation via Metformin-TAK1-IKKα/β-Hsc70 signaling and suggests Metformin as a new activator of CMA for diseases, such as AD, where such therapeutic intervention could be beneficial.