The P2X7 receptor (P2X7 R)is a nonselective cation channel activated by extracellular ATP,and it may trigger the se-cretion of several proinflammatory substances,such as IL-1β, IL-18,TNF-α,and nitric oxide.Several preclinical studies have demonstrated that this receptor participates in inflammation and pain mechanisms,and compounds that modulate the function of this receptor show potential as new anti-inflammatory medicines. Therefore,P2X7 receptors play particularly important physiologi-cal roles and have potential clinical application in inflammation and pain,which proves it a therapeutic target for pain manage-ment.

Objective To investigate the effect of HIF-2a silencing by transfection of siRNA into MG-63 cells un-der hypoxia. Methods HIF-2αexpression level in MG-63 cells under hypoxia was determined by Western Blot. Small in-terfering RNA (siRNA) was used to construct MG-63/siHIF-2α(siHIF-2α)cells and control MG-63/scramble (NC) cells. The expression levels of HIF-2α, Vascular endothelial growth factor (VEGF), p-Erk/ErK and Mcl-1 in MG-63, NC and si-HIF-2αcells was determined by Western Blot. NC and siHIF-2αcells were cultured under hypoxia. Cell viability was as-sessed by MTT assay. Migration was identified by scratch migration assay. Tumor formation was identified by clone formation assay. Nude mouse subcutaneous xenograft model was used to investigate tumor development in vivo. Results Hypoxia im-proved HIF-2αexpression in MG-63 cells in a time-dependent manner (F=2 037.412,P<0.001). HIF-2αexpression un-der hypoxia in siHIF-2αcells was lower than that in NC cells (P<0.01). Cell viability of siHIF-2αcells under hypoxia for 12 h and 24 h were lower than that in NC cells (P<0.05 or P<0.01). The relative width of scratch in siHIF-2αgroup under hypoxia for 12 h and 24 h were larger than that in NC group (P<0.01 or P<0.01). When cell counts reach 1 000-5 000, the clone formation rates of siHIF-2αcells were lower than that in NC cells (P<0.05 or P<0.01). The expression of VEGF, p-Erk/Erk and Mcl-1 protein under hypoxia in siHIF-2αcells was lower than that in NC cells(P<0.01). Tumor sizes, weights and density of siHIF-2α group in nude mice were suppressed compared with those in NC group (P<0.01). Conclusion Blocking HIF-2αsignal pathway warrants its investigation as a potential strategy in osteosarcoma treatment.

Objective Primitive neuroectodermal tumor (PNET) is a rare malignant small round cell tumor .This paper aimed to study the clinical and pathological features of primary pulmonary primitive neuroectodermal tumor . Methods We collected 2 cases of primary pulmonary PNET to review the clinical and pathological features .Immunohistochemical staining was used to detect immune mark-ers, and fluorescence in situ hybridization (FISH) was applied to detect EWS translocation. Results 2 patients were aged 33 years and 17 years.Microscopically, the tumor cell was composed of single small round cells in diffusion or in distribution of sheets or beams , with scant cytoplasm , oval or spindle-shaped nucleus , high mitotic count .Irregular tumor necrosis scattered in the tumor along with visi-ble rosette structure.Immunohistochemical study showed that the tumor cells were positive for CD 99, FLI-1 and Syn, while CKpan, EMA, Desmin, CgA, TTF1, CD34 were negative.EWS/FLI1 translocations were detected positive in both the cases .2 patients died 7 months and 32 months after operation , respectively . Conclusion Primary pulmonary PNET is rare , so the selection of appropriate im-mune markers (CD99, FLI-1, Syn) and FISH for the detection of EWS translocation helps to improve the accuracy of diagnosis .

Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF.Methods The 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF + PD98059 group and an EMF + SB202190 group.Cells in the EMF group were cultured in the electromagnetic field, those in the EMF + PD98059 and EMF + SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally.Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration.The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies.The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit.MTT assay was applied to investigate the proliferation of cells.Results Electromagnetic fields could rapidly induce the activation of p38 MAPK (P ＜ 0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF.The phosphorylation of p38 MAPK was significantly lower in the EMF + SB202190 group than that of the EMF group.After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190.The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190.Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and received 30 minutes of EMF exposure.Conclusion EMF can quickly activate ERK and p38 MAPK pathways to induce cell proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.Moreover, in EMF there is a mutual interference between ERK and p38 MAPK pathways.

Objective To study the changes in miRNAs expression in the exosomes of human umbilical vein endothelial cells(HUVECs) after 60 Co γ-rays expose using microRNA(miRNA) chips and bioinformatics techniques so as to provide new clues to the mechanism of radiation-induced vascular tissue injury and its bystander effects.Methods HUVECs exosomes were collected in the control and 4 Gy irradiated cells by ultra-high-speed centrifugation,and further confirmed using transmission electron microscopy (TEM) and Western blotting of exosomes biomarkers.miRNA microarray was used to analyze miRNA expression profiles of exosomes and cells.Also,real-time quantitative PCR(qRT-PCR) was used to verify differentially expressed miRNAs,and the miRDB and TargetScan were performed to predict the target genes of the differentially expressed miRNAs.Bioinformatics analysis was performed using DAVID,KEGG and other online tools.Results Compared with the control exosomes from non-irradiated HUVECs,miRNA microarray analysis revealed that 5 up-regulated,and 13 down-regulated miRNAs were identified in the exosomes from HUVECs at 0.5 h after 4 Gy-irradiation,and 16 up-regulated and 5 down-regulated miRNAs at 2 h after 4 Gy-irradiation.Moreover,38 and 85 miRNAs were differentially expressed respectively in the HUVECs at 0.5 h and 2 h after radiation.The difference was statistically significant(P<0.01).The results of bioinformatics showed that these miRNAs might exert the radiation-induced bystander effect (RIBE) by regulating MAPK signal pathways,RAS and PI3K-Akt signal pathways.Conclusion The ionizing radiation injury significantly alters the components and expression levels of exosomal miRNAs,which play important roles in regulating the signal pathways in response to radiation.

Aim To look for cold-protective drugs treating cryogenic freezing that may bring great damage to animal physiological system.Methods The protective effect of curcumin on frozen damage and the changes of thyroid function in mice were studied in this study.Quantitative analysis of the changes in survival time and metabolic indexes in mice disposed at(-20±1)℃ was conducted.Mouse serum free triiodothyronine(FT3) and free thyroxine(FT4) contents were detected by ELISA kit.HE staining was used to observe thyroid tissue morphological items.The expression of genes related to thyroid function was assessed via real-time quantitative PCR.Results The intraperitoneal injection of curcumin(12.5~50 mg·kg-1) could remarkably prolong the survival time of mice when exposed to cryogenic freezing.HE staining results displayed a recovered thyroid injury in morphology in the curcumin group, further with a notably improved metabolic indexes and evidently increase in serum FT3 and FT4 levels.The real-time quantitative PCR results indicated that the expressions of sodium iodide symporter(Nis), thyroglobulin(Tg) and thyroid peroxidase(Tpo) were up-regulated, and the expression of thyroid-stimulating hormone receptor(Tshr) was down-regulated.Sodium levothyroxine collabrated with the promoting thyroid effects of curcumin, while propylthiouracil inhibited the effects.Conclusion Curcumin can prolong the survival time of the cryogenic freezing mice, which is closely related to its ability to promote thyroid function.

Objective:To investigate whether interferon-gamma inducible protein-10(IP-10) and interferon-gamma(IFN-?) participated in the human cerebral ischemia injury.Methods:Twenty-one cerebral ischemia specimens, collected from patients died of cerebral infarction, were divided into three groups: less than 7 days, 7-14 days and 15-21 days according to the lasting time of cerebral infarction. The infiltrating of inflammatory cells were observed using HE stain as non-ischemic hemisphere was for controls. Expression of IP-10 and IFN-? in sections both postmortem ischemic hemisphere and non-ischemic hemisphere were detected using immunohistochemistry.Results:In the groups of less than 7 days and 7-14 days large quantity of inflammatory cells were infiltrated in ischemia tissue. Expression of IP-10 in three groups was elevated in the ischemic hemisphere compared with non-ischemic hemisphere(1.74-folds, 1.41-folds and 1.52-folds increases respectively, P0.05).Conclusion:These results showed IP-10 and IFN-? are expressed in human cerebral ischemia. It was suggested that IP-10 and IFN-? involve in cerebral ischemia injury. Moreover, it was also suggested that IP-10 participate in the repair for the later stage of cerebral ischemia injury.

Objective To explore the relationship between lipids profile and body composition in the young and middle?aged adults. Method The relationship between body composition and lipids profile was examined in 642 adults (21-60 years, 178 male, 464 female). According to the result of body composition assessment, they were assigned to three groups: Normal (N, n=272), Obesity (O, n=245), Sarcopenic Obesity (SO, n=125). The lipids profile among three groups and its related factors were analyzed. Results In groups N, O and SO, the levels of total cholesterol (TC), triglycerides (TG), low density lipoprotein?cholesterol (LDL?C) increased gradually (P<0.01), and the high density lipoprotein?cholesterol (HDL?C) level decreased gradually (P<0.01). In a multiple logistic regression analysis, the odds ratio for high levels of TC, TG, LDL?C and low level of HDL?C risk increased gradually in groups N, O and SO [compared to group N, the odds ratio of the four kinds of dyslipidemia in group O were 2.617 (1.117-6.132), 3.549 (1.481-8.503), 4.618 (1.288-16.564), 1.222 (0.529-2.822), respectively, and in group SO were 5.915 (2.512-13.926), 10.430 (4.400-24.722), 9.522 (2.637-34.388) , 4.253 (1.957-9.242) , respectively]. After adjusting for age, sex, waist?to?hip ratio and visceral fat area, the odds ratio for high level of TG risk still increased gradually in group N, O and SO [compared to group N, the odds ratio of group O was 3.565 (1.319-9.632), and of group SO was 8.173 (2.685-24.881)]. Moreover, the odds ratio for high TC and low HDL?C levels of group SO were higher than those of group N [compared to group N, the odds ratio in group SO were 5.658 (1.871-17.111), 6.823 (2.119-21.969) respectively]. With stepwise multivariate logistic regression analysis, for male, the related factors for high levels of TC, TG and low level of HDL?C were high percentage of body fat;sarcopenia, sarcopenia obesity;sarcopenia, respectively. For female, the related factors for high levels of TC, TG and LDL?C were sarcopenia, aging;high percrntage of body fat, aging;sarcopenia, aging, respectively. Conclusion SO was the main risk factor for dyslipidemia in young and middle?aged adults, even more severe than obesity alone and sarcopenia was the risk factor of high TG, low HDL?C levels for male;and the risk factor of high TC, high LDL?C levels for female.

Objective: To observe the conditions of the activating areas in the brain evoked by needling Shenmen (HT 7) acupoint. Methods: Twelve healthy young volunteers were needled at Shenmen (HT 7) acupoint with rotating and twisting techniques. Functional magnetic resonance imaging (fMRI) was adopted to detect the brain activating areas, TR/TE/FA=3560 ms/50 ms/90°. After scanning, the images were dealt with SPM2. Results: Needling Shenmen (HT 7) primarily activated BA2 and BA1 of the gyms postcentralis in right-sided frontal lobe, BA47 of left-sided inferior frontal gyms and BA43 of left-sided gyms postcentralis, and also activated BA40 of inferior parietal lobule in right-sided parietal lobe, BA22 of gyrus temporalis superior in left-sided lobus temporalis, and BA40 of right-sided insula; however, the sham acupoints could not activate these nucleus. Conclusion: Needling Shenmen (HT 7) can activate the relevant functional areas in the brain.

Aim Toevaluatethesynergisticeffectof anti-tumor by the pterostilbene and acetylshikonin act-ing on B16F10 cells and investigate the interrelated mechanisms.Methods Theresearchscreenedandan-alyzed the target-related of pterostilbene and ace-tylshikonin by system-pharmacological methods. The proliferative inhibition rate of B16F10 cells were meas-ured by MTT.The apoptosis in B16F10 cells were proved by both cellular morphological and biochemical methods.The expression of apoptotic genes were as-sessed via RT-PCR.The apoptotic rate and cell cycle were measured by flow cytometry.Melanoma models were established in C57BL/6 mice,and the inhibitory rateoftumorgrowthwasmeasured.Results The14 targets of pterostilbene were closely related to cell cy-cle,acetylshikonin′s 12 targets displayed a relationship with apoptosis,and correlated with p53 signaling path-way.Pterostilbene along with acetylshikonin signifi-cantly inhibited cell proliferation of B16F10 cells in a dose-dependent way and resulted a remarkable syner-gistic effect.The apoptotic rate reached highest with a blocked-cell cycle at G1 phase in the co-treatment group.The RT-PCR results showed that the expres-sions of p53,Bax and p21 were up-regulated and the expressions of Bcl-2,CDK2 and Cyclin E were down-regulated with time.The changes of p53,Bax and Bcl-2 were obvious in combined treated group.All treat-ments in vivo showed different tumor inhibition rates while co-treatment group showed highest.Conclusion Pterostilbenecooperatedwithacetylshikonininhibits the proliferation in B16F10 cells,and activates the p53 signaling pathway to induce the B16F10 cells apoptosis and a cell cycle arrest.