OBJECTIVE:To select the formula,preparation and quality evaluation of water-in-oil microemulsion. METHODS:Water-in-oil micromulsion has been formulated using Volp-5 as surfactant, several short chain alcohols as co-surfactants, and the different oils, pseudoternary phase diagrams have been constructed to evaluate the phase behavior of the formulations of the water-in-oil microemulsion, and the influence of mixing ratio of various elements on the area of microemulsion was also examined. The morphology, particle size, distribution and stability was assessed. RESULTS:The stable water-in-oil microemulsion was made. The microemulsion delivery could be prepared using Volp-5/ethanol/caprylic-capric-triglyceride/water (3∶2∶2∶4)system and no phase separation was observed in three months. CONCLUSION:The formulation provide a convenient means of the oral delivery taking advantage of microemulsion's merit, which is easy to prepare with consistent quality.

BACKGROUND:Previous studies demonstrated that ischemia can be compensated by establishing collateral circulation and microangiogenesis,and exercise training can ameliorate blood supply of ischemic lower limb.However,whether exercise training accelerate establishment of collateral circulation remains poorly understood.OBJECTIVE:To explore the mechanism of exercise training on accelerating microangiogenesis of rat ischemic lower limb.METHODS:Sprague-Dawley rats were randomly divided into the exercise training,model and sham-surgery groups.All animals were established left lower limb ischemia models except those in the sham-surgery group.Rats in the exercise training group were forced running 30 minutes per day at 1 week after model preparation and those in other groups were performed daily activities.The adductor of ischemic lower limb was obtained for the examination of microvessel density and the expressions of vascular endothelial growth factor and basic fibroblast growth factor.Simultaneously,bone marrow-derived endothelial progenitor cells were harvested for detection of microangiogenesis.RESULTS AND CONCLUSION:Number of endothelial progenitor calls,expressions of vascular endothelial growth factor and basic fibroblast growth factor,as well as microvessel density of the exercise training group was obviously greater than those in the model and sham-surgery groups(P<0.01).Compared with the model and sham-surgery groups,in vitro vasculogenesisof bone marrow-derived endothelial progenitor cells in the exercise training group was increased(P<0.05).These findings demonstrated that lower limb ischemia increases microangiogenesis,and exercise training enhances this effect.

Objective To establish a method of neurons and glial cell culture from embryonic Gekko japonicus cerebral cortex. Methods Embryonic(E15) pallium was dissociated and digesting by trypsin.After counting,cells were seeded in culture flask.The glial cells were obtained by using differential adhesion potential combined with successive passage purified methods,and neurons were obtained by using neurobasal medium supplemented with B27.The cells were fixed and analyzed with immunohistochemic assay. Results After tetra-generation,GFAP positive cells were more than 95% in glial cells cultured condition;Neurons grew well in neurobasal medium,and NF and MAP-2 positive signals co-localized on neurons.After cultured for 10 days,the percentage of neurons was more than 95%.Conclusion The methods of isolation,culture and purification for embryonic Gekko japonicus cortical neurons and glial cells were established and it might be a valuable cell model to further investigate the central nerve system in Gekko japonicus.

Aim To investigate the protective effect of RTP1,one of the polysaccharide isolated from Rheum tanguticum,on H2O2 induced apoptosis in IEC-6 cells and its possible mechanism.Methods H2O2(100 mmol?L~-1)was used to induce IEC-6 cell apoptosis.Different doses(10,30,100 mg?L~-1)of RTP1 were administrated before H2O2 was added into IEC-6 cell culture.Cell viability was observed by MTT assay.Reactive oxygen species were measured with laser scanning confocal microscopy(LSCM).DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis,acridine orange staining and flow cytometry.The activation of Caspase-3 was detected with Western blot analysis.Results Following treatment with H2O2 for 2 h,H2O2 induced a significant decrease in cell viability,while DNA ladder was observed and apoptosis percentage was as high as 31.3%.Accumulation of intracellular ROS and increase in Caspase-3 activity were also detected.Pretreatment with RTP1 for 24 h exhibited cytoprotective effects in a dose-dependent manner.RTP1 obviously enhanced cell viability,reduced formation of DNA ladder and significantly reduced the number of cells labeled with Annexin V.The percentage of apoptosis/necrosis cells was markedly decreased to 24.4% and 21.5%,respectively.LSCM showed that RTP1 attenuated the accumulation of ROS.The significant decrease in Caspase-3 activity was detected.Conclusion RTP1 has cytoprotective capacity to antagonize H2O2-induced IEC-6 cell apoptosis and injury,and this effect may be related to decrease ROS and inhibit Caspase-3 activity

Objective:To investigate the effect of NS-398,a selective cyclooxygenase-2(COX-2) inhibitor,on the proliferation and invasion of human colon carcinoma cells in vitro,so as to determine the possibility of COX-2 as a new target for treatment of colon carcinoma.Methods: The expression of COX-2 in colorectal cancer cells(CW-2,COLO-320) was detected by RT-PCR and Western blotting.COLO-320 cell proliferation was measured by MTT after treatment with NS-398.Cell invasion ability was measured using migration and invasion chamber systems.Western blotting assay was used to examine the influence of NS-398 on MMP-2 expression.Results: Our results showed that CW-2,COLO-320 cells expressed COX-2 mRNA and protein.NS-398 inhibited the proliferation of COLO-320 cells in a time-and concentration-dependent manner.Invasion test showed that NS-398 inhibited the migration and invasion of COLO-320 cells.Western blotting revealed that NS-398 inhibited the expression of MMP-2 in COLO-320 cells.Conclusion: The selective COX-2 inhibitor NS-398 can inhibit COLO-320 cell proliferation and invasion,indicating COX-2 may serve as a new target for colon carcinoma treatment.

Objective To investigate the expressions of miR-16,miR-106b,miR-449a,miR-34a and let-7g in peripheral blood plasma of the residents surrounding hot springs with radon in Hebei province.Methods A total of 41 randomly selected residents surrounding hot springs with radon were considered as the radon group,and 46 residents with same living habit but without contact with hot springs were considered as control.The miRNAs in the peripheral blood plasma of these two groups were detected with qRT-PCR.Results The levels of miR-16,miR-106b,miR-449a and let-7g in the radon group were significantly higher than those in control group (Z=-2.278,-3.835,-2.719,-2.721,P＜0.05).Alterations of these miRNAs were associated with radon exposure (t =2.154,3.711,2.319,2.015,P ＜ 0.05) but had no relationship with age,sex,smoking and drinking factors.No significant difference was observed in the plasma levels of miR-34a between the two groups.Conclusions miR-16,miR-106b,miR-449a and let-7g could be applied as potential biomarkers for radon exposure.

prognosis of patients with acute upper gastrointestinal bleeding (AUGIB). Methods A total of 130 patients with AUGIB in our hospital were enrolled in this study from August 2014 to August 2016 . Three kinds of scoring systems including Rockall, Blatchford and AIMS65 were used to evaluate the risk classification of the patients. Patients were followed up for 2 months. The incidence rates of re-bleeding and death within 2 months after admission were observed. Data of prognosis in patients with AUGIB were compared between the three scoring systems in the prognosis of patients with AUGIB, including the risk classification and the prediction accuracy of re-bleeding and death. Results There were no significant differences in the re-bleeding rate and fatality rate between the groups divided by Rockall and Blatchford scoring systems ( P>0.05). According to the grouping results of AIMS65 scoring system, the re-bleeding rate of low-risk group was lower than that in high risk group (1.45%vs. 13.11%, P=0.01), but there was no significant difference in fatality rate between the two groups (P>0.05). The values of area under the curve (AUC) of predicting re-bleeding rates by using Rockall, Blatchford and AIMS65 scoring systems were 0.6258, 0.6910, and 0.7241, and the values of AUC of predicting fatality rates were 0.7031, 0.7969, and 0.7031 by using receiver operating characteristic curve (ROC) analysis. There were no significant differences between them (P>0.05). The values of AUC of predicting re-bleeding rates by using the risk grading calculation by Rockall, Blatchford and AIMS65 scoring systems were 0.6189, 0.6139 and 0.7254 (P>0.05). But the values of AUC of predicting fatality rates were 0.6211, 0.6641 and 0.7695 (P<0.01). Conclusion The operation method of AIMS65 scoring system is simple and convenient, which is applicable to a wide range of patients with AUGIB. In the prediction of re-bleeding and mortality, AIMS65 scoring system has high accuracy and stability, which is worthy of promoting in clinical application.

Objective To investigate the effects of 60Co γ-ray partial radiation on chromosome aberration in human peripheral blood in vitro.Methods The samples of heparinized peripheral whole blood from 3 healthy persons were exposed to 60Co γ-rays at the doses between 0 and 8 Gy with the dose rate of 0.35 Gy/min at the temperature of 37 ℃ ,and then mixed with the unirradiated blood samples of the Microscopy was used to observe the chromosome aberration double ( centromere + centromere) and the biological dose was estimated thereby.ResultsThe amounts of double centromere + centromere were increased along with the dose of irradiation in all groups.The estimated biological dose was higher than the 1/3 of the irradiation dose when the dose was between 0.5 to 2 Gy,and was close to the 1/3 of the irradiation dose when the dose was between 4 to 8 Gy.Conclusion Chromosome aberration can be used as a biomarker in estimation of uneven irradiation.

Objective To investigate 60Co γ-ray induced damage in lymphocytes and the relationship between doses of 60Co γ-ray irradiation and the levels of phosphorylated H2AX and ATM.Methods Cells were irradiated with 60Co γ-rays in the range of 0-8 Gy.The levels of phosphorylated H2AX and ATM were detected by Western blot and FACScan,respectively.The micronucleus(MN)was analyzed by CB method to evaluate DNA damage.Results FACScan results showed the dose-effect relationship of γ-H2AX expression were linear.square at 0.5 h post-irradiation to different doses,and the fitting curve was shown as Y=3.96+11.29D-0.45D2.The level of phosphorylated ATM(p-ATM)was not changed significantly by using the same method.Western blot showed that p-ATM protein expression was significandy increased after irradiation compared with sham.irradiated group.The MN assay which represented DNA damage was sensitive to different doses.Conclusions γ-ray irradiation could induce the phosphorylation of H2AX and ATM,which may play an important role in indicating DNA damage.Both of H2AX and ATM have the potential as sensitive biomarker and biodosimeter for radiation damage.

Objective To analyze the chromosome aberrations induced by partial radiation in human peripheral blood lymphocytes in vitro.Methods Heparinized whole blood samples were exposed to 2 Gy ~(60)Co 7-rays at 37℃ ,and then mixed with non-irradiated blood by different ratio.The slides were prepared after culturing and the unstable aberrations were analyzed.Results The chromosome aberrations had a good relationship with the ratio of irradiated blood.The chromosome aberrations in partial irradiated group were higher than that in the irradiated group.The estimated dose was 1.27 Gy when the ratio was 1 : 1 ,greater than the dose of 1 Gy.The estimated dose was 0.93 Gy when the ratio was 0.5＝1,also greater than 0.5 Gy.But when the ratio was 1:0,the radiation dose was accordant with the estimated dose.Conclusions Chromosome aberrations could be a biomarker for estimating the uneven irradiation.