In this study, we aimed to investigate the influences of conditioned medium from human umbilical vein endothelial cells (HUVEC) on cancer stem cell phenotype of human hepatoma cells. HUVEC and human hepatoma cells (MHCC97H) were cultured, respectively, and then the MHCC97H cells were co-cultured with conditioned medium from HUVEC (EC-CM) with Transwell system. Anti-cancer drug sensitivity, colony-formation, migration/invasion ability, expression of cancer stem cell marker and sphere formation were performed to determine the cancer stem cell phenotype in MHCC97H cells. We found that MHCC97H cells co-cultured with EC-CM exhibited significantly higher colony-formation ability and lower sensitivity of anti-cancer drugs 5-FU and Cis. Transwell assay showed that treatment with EC-CM obviously increased migration and invasion of MHCC97H cells. Moreover, increased sphere forming capability and expression of CD133 in MHCC97H cells were observed after co-cultured with EC-CM. These results suggested that EC-CM could promote cancer stem cell phenotype of hepatoma cells.
Antineoplastic Agents ; pharmacology ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Coculture Techniques ; Culture Media, Conditioned ; Fluorouracil ; pharmacology ; Human Umbilical Vein Endothelial Cells ; chemistry ; Humans ; Liver Neoplasms ; Neoplastic Stem Cells ; cytology ; Phenotype

purpose: to investigate the adhesive properties of renal tubular epithelial cells on matrigel and compared with the following three cases: ischemia、hypoxia and ischemia & hypoxia(I/H).materials and methods: A micropipette aspiration technique was adopted to determine the adhesive mechanics of renal tubular epithelial cells on matrige. results: it showed that the adhesion of renal tubular epithelial cells on matrigel was higher than that of those three model, further more, a different factor was followed by different adhesive mechanic. conclusion: the adhesion of I/H is lower, the ischemia is higher, but all were lower compared with control. It suggested that effect of hypoxia on adhesive properties of renal tubular epithelial cells on matrigel is bigger than that of ichemia.

With the incidence of type 2 diabetes mellitus increasing year after year, the technology of drug screening of type 2 diabetes mellitus progress rapidly, from the level of animal screening to cellular and molecular screening model, from the traditional drug screening technology to high efficient and throughput screening. This paper will summarize the technology of drug screening in the therapy of type 2 diabetes mellitus.
Animals ; Diabetes Mellitus, Type 2 ; drug therapy ; genetics ; Drug Evaluation, Preclinical ; methods ; Humans ; In Vitro Techniques

Matrix metalloproteinases (MMPs) are endocellular proteolytic enzymes. They are so named because they need Ca2+, Zn2+ and other metal ions as their cofactors. MMPs play an important biological role in regulating the formation, remodeling and degradation of extracellular matrix and participate in various physiological and pathological processes of cells. Bone marrow-derived mesenchymal stem cells (BMSCs) are a kind of pluripotent stem cell which has the ability to self-renew and differentiate into functional cells. Meanwhile, they can respond to the damage signals and migrate to injured site for tissue repair and regeneration. MMPs and their inhibitors TIMPs affect the differentiation and migration of BMSCs. This article reviews the roles of MMPs in differentiation and migration of BMSCs.
Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cell Movement ; physiology ; Humans ; Matrix Metalloproteinases ; physiology ; Mesenchymal Stromal Cells ; cytology

BACKGROUND:Cel cryopreservation is required for clinical use of stem cel s, and the current process of cryopreservation however may be harmful to cel viability, pluripotency and differentiation capacity.
OBJECTIVE:To explore the effect of fructose and dithiothreitol on pluripotency and osteogenesis of cryopreserved bone marrow mesenchymal stem cel s.
METHODS:Bone marrow mesenchymal stem cel s were isolated from the bone marrow of Sprague-Dawley rats and pretreated with fructose (200μmol/L), dithiothreitol (500μmol/L) or combined components before cryopreservation. Then the cel s were cryopreseved for 6 months and the morphology of cel s was observed by inverted microscopy. The cel viability was evaluated by MTT, and real-time PCR was used to detect the mRNA expression of Nanog, OCT4 and Sox2. Alkaline phophatase activity assay and alizarin red staining were utilized to detect the osteogenic capacity of bone marrow mesenchymal stem cel s.
RESULTS AND CONCLUSION:Images captured by inverted microscopy showed no significant difference in cel morphology between groups. The MTT results indicated that fructose and combined pretreatment could promote the cel viability of bone marrow mesenchymal stem cel s after cryopreservation, while the real-time PCR results demonstrated that dithiothreitol significantly facilitated the expression of Naogo and Sox2 in bone marrow mesenchymal stem cel s. Moreover, ALP activity assay and alizarin red staining confirmed the positive effects of fructose, dithiothreitol and combined pretreatment on osteogenic capacity of bone marrow mesenchymal stem cel s after cryopreservation, and the best effects were found after pretreatment with dithiothreitol and combined components. Overal , these findings indicate that fructose pretreatment is beneficial for cel viability of cryopreseved bone marrow mesenchymal stem cel s, and dithiothreitol contributes to maintaining the pluripotency and osteogenesis capacity of cryopreseved bone marrow mesenchymal stem cel s.

Objective To investigate the effect of dexmedetomidine combined with butorphanol conventional therapy on sustaining sedation of intensive care unit (ICU) patients.Methods Sixty critically ill patients in Binzhou Central Hospital from June to September in 2014 were randomly divided into experimental group and control group, 30 cases in each group. In the control group, 0.8 mg/L dexmedetomidine hydrochloride injection (400μg with addition of 46 mL normal saline to form 50 mL solution) was intravenously infused continuously at a speed of 0.4μg·kg-1·h-1 by a micro-pump to induce analgesia and sedation; while in the experimental group, dexmedetomidine combined with 200 mg/L butorphanol (10 mg plus 40 mL normal saline to form 50 mL solution) was given for intravenous infusion by a micro-pump with a speed of 0.01 mg·kg-1·h-1 to maintain analgesia and sedation for 48 hours whose required Ramsay score in both groups was 3 - 5. Before and after treatment, the changes of heart rate (HR), respiratory rate (RR), mean arterial pressure (MAP), arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), and the pulse oxygen saturation (SpO2) of both groups were observed. The dosage of dexmedetomidine used for maintenance of required analgesia and sedation and FPS (facial expression) grading and Ramsay score were compared respectively between the two groups, and the clinical efficacy of the two groups were evaluated.Results After treatment, the HR, MAP, RR in both groups were significantly lower, and PaO2 and SpO2 were significantly higher than those before treatment, and the degrees of improvement in the above indexes of the experiment group were superior to those of the control group [HR (bpm): 84.58±12.43 vs. 118.62±14.21, MAP (mmHg, 1 mmHg = 0.133 kPa): 82.35±12.12 vs. 92.35±12.32, RR (times/min): 25.42±3.98 vs. 32.87±5.12, PaO2 (mmHg): 95.21±10.55 vs. 75.18±8.57, SpO2: 0.981 4±0.102 8 vs. 0.954 7±0.093 8, allP < 0.05]. The total therapeutic effect in experiment group was significantly higher than that in control group [93.3% (28/30) vs. 76.7% (22/30),P < 0.05]. The dexmedetomidine dosage used in the experiment group was much less than that in the control group (μg/d: 412.12±23.18 vs. 520.05±15.68,P < 0.05). The FPS score in the experiment group was obviously lower than that in the control group (1.48±0.16 vs. 2.52±0.74,P < 0.05).Conclusion In comparison, to achieve sustained and required analgesic and sedative effect for ICU patients by combined use of dexmedetomidine and butorphanol, the dosage of dexmedetomidine used is less than dexmedetomidine applied alone, in addition, the combined use can achieve better Ramsay grading, steady blood pressure and excellent effect.

In our study, a two-phase culture system was developed to acquire large amount of CD41+ and polyploidy cells. Human mobilized peripheral blood CD34+ (PB CD34+) cells were first cultured in expansion medium (Cocktail or CC100 medium) for 3,4,5 or 6 days, and then cultured in megakaryocytic differentiation medium containing TPO and SCF for additional 7, 8 or 9 days. Cell expansion, morphology, CD41+ cell percentage and DNA content were investigated to evaluate the protocol. The result showed that more CD41+ and polyploidy cells could be obtained following the two-phase culture with Cocktail medium than with CC100. Moreover, with 3 days expansion in Cocktail medium plus 7 days in differentiation medium, the initial CD 34+ cells obtained 16-fold expansion of CD41+ cells and 3-fold expansion of polyploidy cells, such obtained level being significantly higher than that of culturing cells with only one step in TPO or TPO+SCF. We conclude that with the two-phase culture system, PB CD34+ cells can expand and differentiate to more CD41+ and polyploidy cells than those cultured only in accordance to the one-stage culture protocol, so a new and highly efficient megakaryocyte differentiation model for megakaryocyte and platelet related researches is provided already.
Antigens, CD34 ; blood ; Blood Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; physiology ; Colony-Stimulating Factors ; physiology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Megakaryocytes ; cytology ; Stem Cells ; cytology

Bone marrow derived mesenchymal stem cell (BMSC) is one of the crucial cell types which plays roles in wound healing of tissues. In the last decades, it was believed that BMSCs promoted wound healing by differentiating into multiple lineages and placing the wounded tissues. In recent years, a new viewpoint arose from evidences that the paracrine effect of BMSCs might play a more important role in the process of wound healing than differentiation. Understanding the role of BMSCs paracrine in wound healing would be vital to clarify the mechanism how BMSCs take part into the process of wound healing. In this paper, we review the new research processes of BMSCs paracrine in wound healing of tissues.
Animals ; Bone Marrow Cells ; cytology ; physiology ; Cell Differentiation ; physiology ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology ; Paracrine Communication ; physiology ; Wound Healing ; physiology

Mesenchymal stem cells (MSCs) are pluripotent stem cells with high self-proliferation and multidirectional differentiation potential. They also have other functions including immune regulation, paracrine and so on, playing an important role in repairing injured tissues. In recent years, a lot of research has been done on how MSCs promote skin injury repair, and a lot of progress has been made. Compared with direct injection of MSCs in the wound area, some special treatments or transplantation methods could enhance the ability of MSCs to repair skin injury. This paper mainly discusses the role of MSCs in skin injury repair and technical ways to improve its repairing capacity, and discusses the existing problems in this field and prospects for future research directions.
Cell Differentiation ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stem Cells ; Skin

Objective To study the effects of osteopontin (OPN) on the nuclear mechanics of bone marrow-derived mesenchymal stem cells (BMSCs) as well as its involved mechanisms. Methods The BMSC migration was evaluated using the Transwell assay. An atomic force microscope (AFM) was used to determine the elastic modulus of the BMSC nucleus and analyze the changes in the nuclear mechanics of the BMSCs after treatment with OPN. The activation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase1/2 (ERK1/2) was measured by Western blot. The role of the FAK-ERK1/2 signaling pathway in mediating the OPN-affected BMSC nuclear mechanics was investigated by employing a specific inhibitor. RT-PCR and Western blot were used to detect the expression of Lamin A/C at mRNA and protein levels in the BMSCs, respectively. Results The elastic modulus of the BMSC nucleus exhibited a significant decrease after OPN treatment compared with that of the control group. OPN could upregulate the phosphorylation level of FAK and ERK1/2, but the inhibitor of FAK or ERK1/2 restored the OPN-decreased elastic modulus of the BMSC nucleus and inhibited the BMSC migration significantly. After treatment with OPN, the expression of Lamin A/C in the BMSCs reduced significantly, and such a reduced expression could be suppressed by the inhibitor of FAK or ERK1/2. Conclusions OPN could probably downregulate the expression of Lamin A/C of the BMSCs via the FAK-ERK1/2 signaling pathway, decrease the stiffness of the BMSC nucleus, and promote the migration of the BMSCs. The research outcomes provide the experimental evidence for further understanding the mechanism of the OPN-regulated BMSC migration and its potential clinical application.