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Objective To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on macrophage polarization under hypoxia. Methods hUC-MSCs were obtained by explants adherent culture and cultured under normoxia (21% O2) and hypoxia (5% O2). The multi-directional differentiation of hUC-MSCs was observed by osteogenic and adipogenic differentiation induction. Live/death staining was performed to detect the cell viability, and ELISA was executed to detect the protein content in supernatant of hUC-MSCs. Transwell chamber was employed to co-culture the hUC-MSCs cultured under normoxia and hypoxia and macrophage (THP-1) stimulated by lipopolysaccharide (IPS), then the polarization of THP-1 was detected by immunofluorescence, and the secretions of inflammatory factor and anti-inflammatory factor of THP-1 were detected by ELISA. Results hUC-MSCs cultured under hypoxia showed the ability of osteogenic and adipogenic multi-directional differentiation. Live/death staining showed the high cell viability of hUC-MSCs cultured under normoxia and hypoxia. The expression levels of prostaglandin E2 (PGE2) and indoleamine 2,3-dioxygenase (IDO) were significantly higher in the hUC-MSCs cultured under hypoxia than in those cultured under normoxia. hUCMSCs cultured under hypoxia promoted the polarization of THP-1 to M2, obviously reduced the expression of TNF-α and IL-1β, and increased the expression of IL-10 significantly. Conclusion hUC-MSCs cultured under hypoxia may promote the polarization of THP-1 to M2 and improve the viability of anti-inflammatory. DOI: 10.11855/j.issn.0577-7402.2016.07.01

Objective To study the therapeutic effect of exosome derived from lipopolysaccharides (LPS) priming mesenchymal stem cells (MSCs) for diabetic wound healing. Methods Human umbilical cord MSCs were treated with LPS (100ng/ml) for 2 days, the supernatant were then collected, and exosomes were harvested by density gradient centrifugation and identified. Diabetic cutaneous wounds were prepared and the animals were divided into the following three groups: control group, untreated MSCs derived exosome (un-exosome) treatment group and LPS primed MSCs derived exosome (LPS-exosome) treatment group. Exosomes (60μg) were injected dispersively into the wound edge daily for 10 days. After treatment, the therapeutic results were evaluated by gross observation of the wounds, the expression levels of inflammation related factors and macrophage subtype markers in the injured sites were detected by qRT-PCR at day 3, 7 and 14 after treatment. Results Compared with control group, the diabetic wound healing was obviously improved in LPS-exosome treatment group after treatment for 7 and 14 days, with faster wound close, depressed expression of pro-inflammatory factors IL -1, IL -12 and M1 macrophage surface marker iNOS, and up-regulation of anti-inflammatory factors IL-10, TGF-βand M2 macrophage surface marker CD163, the differences were significant (P

Objective To evaluate the temporary hypoglycemic effect of adipose mesenchymal stem cells (AD-MSCs) on type 2 diabetic rats, and further discuss the mechanisms of AD-MSCs regulating blood glucose through hepatic glycometabolism. Methods The type 2 diabetic rat model was established by intraperitoneal injection of a single small dose of streptozotocin (STZ, 25mg/kg) after a high-fat diet for 8 weeks. The content of blood glucose was determined by tail vein bleeding at indicated time points (0h, 3h, 6h, 12h, 24h) after infusion of AD-MSCs, concurrently the rats were executed and the liver tissues were collected (diabetes group, n=5, normal group, n=3) for detecting the mRNA and protein expressions of glucokinase (GK), pyruvate kinase (PK) and phosphofructokinase (PFK) (the key enzyme of glycolysis) by real-time qPCR and Western blotting. Results The infusion of AD-MSCs significantly decreased blood glucose in a short period [30.55±1.49mmol/L(0h), 18.90±0.85mmol/ L(3h), 23.70±1.13mmol/L(6h), 21.90±1.00mmol/L(12h), respectively]. PCR results showed that the expressions of GK and PFK increased in the liver tissue of type 2 diabetic rats 3h after AD-MSCs infusion, and the expression of PK increased significantly 6h after AD-MSCs infusion (P

Diabetes is a chronic metabolic disorder caused by relative or absolute insulin deficient or reduced sensitivity of target cells to insulin. Mesenchymal stem cells (MSCs) are adult stem cells with multiple differentiation potential, self-renewable and immunoregulatory properties. Accumulating evidences from clinic or animal experiments recent years showed that MSCs infusion could ameliorate hyperglycemia in diabetes. The research progress of MSCs in diabetes treatment is summarized and a corresponding perspective is herewith proposed in present paper. DOI: 10.11855/j.issn.0577-7402.2016.07.16

Objective To investigate the effect and mechanism of bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) on pro-inflammatory cytokines-induced apoptosis of insulinoma cells (INS-1). Methods The conditioned medium containing various cytokines and growth factors was collected and condensed from the supernatants of culture medium of BMSC of rats at passage three. After exposed to the medium with a pro-inflammatory cytokines cocktail containing TNF-α, IL-1β and IFN-γ for 48 hours, INS-1 cells were cultured with BMSC-CM in different concentrations for another 12 hours. INS-1 cells were divided into the following five groups: control group, pro-inflammatory cytokines-treated group, 1× BMSC-CM-treated group, 2× BMSC-CM-treated group and 4× BMSC-CM-treated group. The survival rate of INS-1 cells was assessed by CCK8 assay. The incidence of cell apoptosis was measured using Annexin V/PI staining. Intracellular reactive oxygen species (ROS) production was determined with dichlorofluorescein diacetate (DCFH-DA). Results Compared with the control group, prolonged exposure to pro-inflammatory cytokines induced significant decrease in the viability of INS-1 cells (75.84%±1.53%) and a great increase in the incidence of cell apoptosis (17.23%±1.77%), and also in intracellular ROS level (34.3%±0.80%) with statistically significant difference (P

Objective To explore the association between high-risk human papillomavirus (HR-HPV) viral load and pathological grades of cervical intraepithelial neoplasia (CIN) and cervical cancer. Methods A total of 1248 patients from General Hospital of PLA, who underwent colposcopy and surgery due to cervical lesions between Jan. 2006 and Aug. 2011 were enrolled in this study, and they were divided five groups: cervicitis, CIN Ⅰ, CIN Ⅱ-Ⅲ, stage Ⅰ cervical cancer and stage Ⅱ cervical cancer. HR-HPV viral load (RLU/CO) was determined by the Hybrid Capture Ⅱ (HCⅡ) system, and they were categorized into five groups: 0-0.99, 1.00-9.99, 10.00-99.99, 100.00-999.99, ≥1000.00. The mean value and standard deviation of different HR-HPV viral load in the patients with cervicitis or with CIN Ⅰ, CINⅡ-Ⅲ, stage Ⅰ cervical cancer or stage Ⅱ cervical cancer were compared, and the correlation of HR-HPV viral load and pathogenesis of cervical lesions was analyzed. Results HPV viral loads were significantly higher in CINⅠ(842.1±983.9), CINⅡ-Ⅲ (690.1±795.0), stage Ⅰ cervical cancer (893.1±974.2) and stage Ⅱ cervical cancer (699.5±908.3) patients than in cervicitis patients (274.2±613.6, P < 0.05), and the HPV viral loads in CINⅠ(842.1±983.9) and stage Ⅰ cervical cancer patients were higher than those in CINⅡ-Ⅲ patients (P < 0.05). When HR-HPV viral load was ≥100RLU/CO, the risk of CIN and cervical cancer increased with the increase in viral load, but there was no correlation between the viral load and pathological grades of cervical lesions. In the patients with stage ⅠB-Ⅱ cervical squamous cell carcinoma, when the HR-HPV viral load was ≥100RLU/CO, the risk of lymph node metastasis increased (P < 0.05), and the number of patients with maximum diameter of the cervical tumor ≥4cm also increased (P < 0.05). However, the HR-HPV viral load was not correlated with patient age, pathological type of the lesion, depth of cancer cell invasion into cervical muscle layer and lymphatic invasion. Conclusions HR-HPV viral load is closely correlated with genesis of CIN and cervical carcinoma. The higher the viral load, the greater the risk of CIN and cervical cancer is, and it is correlated with the clinical features of cervical squamous cell carcinoma.