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 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.

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.

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 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.

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 establish the reference intervals of total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C)and low density lipoprotein (LDL-C)in the healthy population in Hengshui region.Methods 16 403 healthy individuals were collected from the various counties and cities in this region and detected the above 4 blood lipid indexes (TC,TG,HDL-C, LDL-C)by adopting the automatic biochemical analyzer.The subjects were grouped according to different genders and ages and the detection results were performed the statistical analysis.Results TC 3.17-7.20 mmol/L;TG 0.08-3.16 mmol/L;HDL-C 0.94-2.30 mmol/L;LDL-C 1 .14 -4.59 mmol/L.Conclusion The differences of blood lipid detection results exist in males and fe-males in Hengshui region,the results are different among different age groups,it is suggested that different genders and different age groups should establish their own normal reference intervals in order to provide the diagnostic basis for clinic.

Objective To study and compare the clinical efficacy between intravitreal conbercept injection and (or) macular grid pattern photocoagulation in treating macular edema secondary to non-ischemic branch retinal vein occlusion (BRVO).Methods Ninety eyes of 90 patients diagnosed as macular edema secondary to non-ischemic BRVO were enrolled in this study.Forty-eight patients (48 eyes) were male and 42 patients (42 eyes) were female.The average age was (51.25 ± 12.24) years and the course was 5-17 days.All patients were given best corrected visual acuity (BCVA),intraocular pressure,slit lamp with preset lens,fluorescence fundus angiography (FFA) and optic coherent tomography (OCT) examination.The patients were divided into conbercept and laser group (group Ⅰ),laser group (group Ⅱ) and conbercept group (group Ⅲ),with 30 eyes in each group.The BCVA and central macular thickness (CMT) in the three groups at baseline were statistically no difference (F=0.072,0.286;P=0.930,0.752).Patients in group Ⅰ received intravitreal injection of 0.05 ml of 10.00 mg/ml conbercept solution (conbercept 0.5 mg),and macular grid pattern photocoagulation 3 days later.Group Ⅱ patients were given macular grid pattern photocoagulation.Times of injection between group Ⅰ and Ⅲ,laser energy between group Ⅰ and Ⅱ,changes of BCVA and CMT among 3 groups at 1 week,1 month,3 months and 6 months after treatment were compared.Results Patients in group Ⅰ and Ⅲ had received conbercept injections (1.20 ± 0.41) and (2.23 ± 1.04) times respectively,and 6 eyes (group Ⅰ) and 22 eyes (group Ⅲ) received 2-4 times re-injections.The difference of injection times between two groups was significant (P＜0.001).Patients in group Ⅱ had received photocoagulation (1.43 ±0.63) times,9 eyes had received twice photocoagulation and 2 eyes had received 3 times of photocoagulation.The average laser energy was (96.05 ±2.34) μV in group Ⅰ and (117.41 ±6.85) μV in group Ⅱ,the difference was statistical significant (P=0.003).BCVA improved in all three groups at last follow-up.However,the final visual acuity in group Ⅰ and group Ⅲ were better than in group Ⅱ (t=4.607,-4.603;P＜0.001) and there is no statistical significant difference between group Ⅲ and group Ⅰ (t=-0.802,P=0.429).The mean CMT reduced in all three groups after treating for 1 week and 1 month,comparing that before treatment (t=-11.855,-10.620,-10.254;P＜0.001).There was no statistical difference of CMT between group Ⅰ and Ⅲ at each follow up (t=0.404,1.723,-1.819,-1.755;P=0.689,0.096,0.079,0.900).CMT reduction in group Ⅰ was more than that in group Ⅱ at 1 week and 1 month after treatments (t=-4.621,-3.230;P＜0.001,0.003).The CMT in group Ⅲ at 3 month after treatment had increased slightly comparing that at 1 month,but the difference was not statistically significant (t=1.995,P=0.056).All patients had no treatment-related complications,such as endophthalmitis,rubeosis iridis and retinal detachment.Conclusions Intravitreal conbercept injection combined with macular grid pattern photocoagulation is better than macular grid pattern photocoagulation alone in treating macular edema secondary to non-ischemic BRVO.Combined therapy also reduced injection times comparing to treatment using conbercept injection without laser photocoagulation.

Objective To investigate the risk factors for traumatic lumbar punctures in children with acute lymphoblastic leukaemia. Methods 132 children with acute lymphoblastic leukemia totally received 2634 lumbar punctures. The basic data on age, sex, body mass index (BMI), platelet count, interval between two punctures, and presence or absence of ultrasound-guided procedure were collected and analyzed. The risk factors for traumatic lumbar puncture were identified by logistic regression. Results The risk for traumatic lumbar puncture was higher in children younger than 1 year, and it was relatively lower in those aged 1 to 10 years. The risk for traumatic lumbar puncture was slightly higher in children with a BMI index of more than 95. The longer the interval between two punctures, the lower the risk. If lumbar puncture was guided under ultrasound or radiographic images, the risk was much smaller. Conclusions Age of younger than 1 year, BMI index of more than 95, shorter interval between two punctures, and direct puncture can increase the risk for traumatic lumbar puncture.