Objective To derive hematopoietic stem cells with functional properties of hematopoietic reconstitution from murine embryonic stem (ES) cells. Methods ES-D3 cells by formation of the day-4 embryoid bodies (4dEBs) were induced into hematopoietic stem cells by co-culture with murine bone marrow endothelial cell-conditional medium (mBMEC-CM) and the fetal liver stromal cell-conditional medium (FLSC-CM). This experiment was designed to 4 groups (mBMEC-CM + FLSC-CM group, mBMEC-CM group, FLSC-CM group, and the control group). Results The total cell numbers, CD34 + cell numbers, and colony numbers formed in the mBMEC-CM + FLSC-CM group were the highest among the 4 groups. The cells in the mBMEC-CM + FLSC-CM group resumed the hematopoietic system of the mice after being transplanted with the inducing cells. Conclusion The culture condition combing mBMEC-CM with FLSC-CM can promote murine ES cells differentiating into hematopoietic stem cells with functional properties of hematopoietic reconstitution.

Objective To explore the protective effects of lycium barbarum polysaccharides(LBP) on fibroblast in vitro irradiated by ultraviolet A(UVA). Methods Taking the primary cultured fibroblast as objects, the fibroblast was irradiated by UVA ( irradiation intensity: 2.4 J/cm2). The fibroblast was randomly divided into six groups, control group, UVA radiated group and four protective groups(0.1 mg/ml LBP, 0.2 mg/ml LBP, 0.4 mg/ml LBP and 0.8 mg/ml LBP). The activities of cell proliferation were measured by MTT methods. The contents of MDA, the activities of SOD in the fibroblasts, and the activities of LDH in the supernatants of fibroblasts were determined by biochemical methods. Results The fibroblasts were irradiated by UVA (irradiation intensity: 2.4 J/cm2),the activities of cell proliferation was decreased,the activities of SOD was decreased too, the content of MDA and LDH increased. Compared with UVA irradiated group,in the given concentration,LBP could improve the activities of cell’s proliferation,improve the activities of SOD and decrease the contents of MDA in the cell, and decrease the content of LDH in the supernatants of cells significantly (P

AIM: To purify murine yolk sac endothelial cells (mYS-EC) and investigate the cytokines mRNA expression in mYS-EC. METHODS: The murine yolk sacs were digested with 0.1% collagenase, resuspended in DMEM and counted after digestion and centrifugation. The yolk sac adherent cells were cultured in DMEM containing 15% FBS with 10% mBMEC-CM or 5?g/L VEGF, ECGF and bFGF. The phagocytose function and expression of vWF were evaluated via particle phagocytosis and immunohistochemistry method. Atlas cDNA expression array was used for analysis of cytokine expression in mYS-EC. RESULTS: Colonies consisting of pure yolk sac endothelial cells were obtained in liquid culture system containing 15% FBS and 10% mBMEC-CM or 5?g/L VEGF, ECGF and bFGF. For complete purification of the endothelial cells, subsequent passage was also necessary. Cellular cord formed during passage culture. The endothelial cells were round or oval sharp in morphology, positive in phagocytosis and factor VIII related antigen (von Willebrand's Factor,vWF). The mRNA expressions of cytokines, such as TGF-?2, TNF-?, IFN-?, FL, BMP-4, MIP-1?, BMP-2A, FLT2, endothelin 2, thymosin ?10, IL-6, IL-13, IL-9, SCYA5 and ACBP were detected in mYS-ECs. CONCLUSION: mYS-EC was purified and expanded in vitro . The mRNA expression of 15 kinds of cytokines was detected in mYS-ECs by Atlas arrays.

Objective To establish the canine model of esophageal variees caused by portal hyper-tension. Methods The model was established in 12 dogs with a side-to-side portacaval shunt, an ameroid constrictor around the portal vein and double ligation and cross suture of the cephalic part of the inferior vena cava (IVC), and the development of esophageal varices was confirmed 6 weeks later by endoscopy and portal vein angiography. Results The mean pre- and postoperative portal pressure were (11.0 ± 1.1) mm Hg and (22. 9 ± 1.2) nun Hg, respectively (P =0. 010). Endoscopy detected mild to moderate esophageal varices in all dogs, which was confirmed by portal vein angiography, and varices was also seen in abdominal wall. Conclusion Canine model of esophageal varices induced by portal hypertension can be established with the procedure.

The present study investigated the effects of the serum-free conditioned media of the bone marrow endothelial cells on CFU-F for potential mechanisms upon which hematopoiesis may be regulated by them within the bone marrow microenvironment. After obtaining the serum-free conditioned media of human and murine purified bone marrow endothelial cells (hBMEC-CM and mBMEC-CM) in vitro, MW > 10 kD, 3 - 10 kD and < 3 kD components were sifted out from these media by means of serial ultrafiltration. Assays of CFU-F were performed to test the effects of BMEC-CM and their ultrafiltrated components. The results showed that every one of hBMEC-CM, mBMEC-CM and their MW < 3 kD components exerted a suppressive effect on the proliferation of corresponding CFU-F but MW > 10 kD and 3 - 10 kD components. The BMEC-CM and MW < 3 kD components decreased the number as well as the size of CFU-F. There were the markedly negative concentration-dependent relations between the concentrations of MW < 3 kD component and the numbers of CFU-F. These findings suggested that bone marrow endothelial cells in culture could secrete at least a humoral factor (molecular weight less than 3 kD) which has an inhibitory effect on the growth of CFU-F.

Animals ; Blood Platelets ; cytology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Culture Media, Conditioned ; Cyclophosphamide ; Female ; Fibroblasts ; cytology ; Male ; Megakaryocytes ; cytology ; Mice ; Stem Cells ; cytology ; Thrombocytopenia ; chemically induced ; therapy

Objective To investigate the function and characteristics of histamine receptors on the hemopoietic progenitor cells.Methods BDF1 mice (both male and female), inbred at our university, aged 8-12 weeks, weighing 20-24 g, were used in this study. Bone marrow cells were incubated for 1 hour at 37℃ with 2-AT (H1 receptor agonist) or impromidine (H2 receptor agonist) alone, or in combination with the antagonists pyrilamine or cimetidine respectively. Control experiment was performed in Dulbecco's modified Eagle's Medium (DMEM) alone. Cells treated with different drugs were performed by colony forming unit-granulocyte-macrophage (CFU-GM) and colony forming unit-megakaryocyte (CFU-Meg) assay.Results When bone marrow cells were treated with 10-8 mol/L to 10-5 mol/L of 2-thiazolylethylamine (2-AT) which had no influence on CFU-GM and CFU-Meg proliferation, 10-8 mol/L to 10-5 mol/L of impromidine could increase the number of CFU-GM and CFU-Meg colonies. The effects of H2 receptor agonists on CFU-GM, CFU-Meg could be antagonized by H1 receptor agonist.Conclusions Our findings suggest the existence of histamine H1 and H2 receptors on the hemopoietic progenitor cells and the antagonism between two different histamine receptor subtypes on the proliferation of CFU-GM and CFU-Meg.

Spinal nerve injury causes mechanical allodynia and structural imbalance of neurotransmission, which were typically associated with calcium overload. Storeoperated calcium entry (SOCE) is considered crucial elements-mediating intracellular calcium homeostasis, ion channel activity, and synaptic plasticity. However, the underlying mechanism of SOCE in mediating neuronal transmitter release and synaptic transmission remains ambiguous in neuropathic pain. Neuropathic rats were operated by spinal nerve ligations. Neurotransmissions were assessed by whole-cell recording in substantia gelatinosa. Immunofluorescence staining of STIM1 with neuronal and glial biomarkers in the spinal dorsal horn. The endoplasmic reticulum stress level was estimated from qRT-PCR. Intrathecal injection of SOCE antagonist SKF96365 dose-dependently alleviated mechanical allodynia in ipsilateral hind paws of neuropathic rats with ED 50 of 18 μg. Immunofluorescence staining demonstrated that STIM1 was specifically and significantly expressed in neurons but not astrocytes and microglia in the spinal dorsal horn. Bath application of SKF96365 inhibited enhanced miniature excitatory postsynaptic currents in a dosage-dependent manner without affecting miniature inhibitory postsynaptic currents. Mal-adaption of SOCE was commonly related to endoplasmic reticulum (ER) stress in the central nervous system. SKF96365 markedly suppressed ER stress levels by alleviating mRNA expression of C/ EBP homologous protein and heat shock protein 70 in neuropathic rats. Our findings suggested that nerve injury might promote SOCE-mediated calcium levels, resulting in long-term imbalance of spinal synaptic transmission and behavioral sensitization, SKF96365 produces antinociception by alleviating glutamatergic transmission and ER stress. This work demonstrated the involvement of SOCE in neuropathic pain, implying that SOCE might be a potential target for pain management.

Objective To detect the spatial features of measles in China by means of spatial statistical analysis. Methods Data of prefecture-level measles cases and incidence from 2005 to 2014 were collected from the China Information System for Disease Control and Prevention. Information collected from the system included demographic characteristics, spatial distribution information, and diagnostic reports. Cases of unconfirmed measles and those with unknown address were ruled out. Cases from Hong Kong, Macao, Taiwan, and foreign countries were not included in this study. Maps were obtained from geographical boundary data at prefecture level from the Chinese Center for Disease Control and Prevention and demographic data from the National Bureau of Statistics. Based on different measures of measles elimination, we divided the data from 2005 to 2014 into three stages: stage 1 (2005-2008), stage 2 (2009-2012), and stage 3 (2013-2014). ArcGIS software was used to describe the spatial distribution and for global and local spatial autocorrelation analysis. Results The total number of confirmed measles cases reported in the system was 650 222, with average incidence 0.46/100 000. The highest reported incidence was in 2008 (9.95/100 000) and the lowest in 2012 (0.46/100 000). Average incidences for stages 1, 2, and 3 were 8.87/100 000, 1.99/100 000 and 2.96/100 000, respectively. Global Moran's I coefficients from 2005 to 2014 were 0.31, 0.08, 0.36, 0.56, 0.26, 0.48, 0.34, 0.20, 0.29 and 0.52, respectively;all were significant (P<0.05). Average incidences for high-high (H-H) clusters in 2005-2008, 2009-2012, and 2013-2014 were 33.02/100 000, 7.06/100 000, 11.91/100 000, respectively. Western China had high-value clustering consistently throughout all three periods; however, the number of prefectures covered by high-value clusters and discrepancy in the incidence between western and eastern regions were low. Northeast and northern China had H-H clustering in stages 2 and 3. Conclusion In this study, measles incidence was spatially autocorrelated at the prefecture level from 2005 to 2014. Although China has made great progress in the elimination of measles, H-H clusters were consistently present. A need remains in China for targeted measles prevention and control measures.