Objective To discuss the technology of modest isolating and enriching the intestinal epithelial stem cells. Methods Mouse intestinal epithelial cells were stained by Rhodamine 123 (Rho), sorting the Rhodamine 123 low staining cell population ( Rholow ) and Rhodamine 123 strong straining cell population ( Rhobri ) by fluorescence activated cell sorting(FACS) in flow cytometer; Detecting the musaashi-1 and p-glycoprotein 1 (p-g-p1) mRNA expression of two groups by RT-PCR; Analyzing the cell cycle and the percentage of the musashi-1 positive cells by flow cytometry. Results The intestinal epithelial cells were divided into three groups, Rhodamine 123 low staining cells( 12. 34% of total cells), Rhodamine 123 middle staining cells (45.26% of total cells) and Rhodamine 123 strong staining cells ( 41. 40% of total cells). The Rholow cell fraction and Rhobri cell fraction were isolated successfully. Both of musashi-1 and p-g-p1 mRNA were strongly expressed in Rho1ow cell fraction, and Rhobri cell fraction little expressed p-g-p1 mRNA. Most of Rho1ow cells were in G0/G1 phase, and the musashi-1 positive cells were about 10.37% of total cells in this fraction. Conclusion The intestinal epithelial stem cells can be modestly isolated and enriched by Rhodamine 123 staining.

0.05).Conclusion TP may have protective effects on chronical hypoxia induced polycythemia in rats.

Objective To observe the effects of MSH2 gene re expression on estrogen-induced apoptosis of colon cancer cells LOVO,and to explore its mechanisms.Methods According to different plasmid and whether with estradiol intervention,colon cancer LOVO cells were divided into empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,estrogen receptor (ER) β with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group,and received corresponding treatment.The expression of MSH2,ERβ protein and apoptosis related caspase 3 protein were detected by Western blotting.Cell viability was measured by cell counting kit-8.Cell DNA fragments of each group were isolated with apoptosis DNA fragments isolation kit.And the DNA ladder was observed.The rate of apoptosis was detected by flow cytometer.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between the two groups.Results After transfection,the expression of the MSH2 and ERβ at protein level in LOVO cells significantly increased and neither of their expression was effected by estradiol.The expression levels of caspase 3 cleavaged active fragments of ERβ with estradiol group and ERβ with MSH2 and ethanol group were higher than other groups,and there was no significant difference between these two groups.The LOVO cell viability of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 1.72 ±0.25,1.74 ± 0.31,1.77 ± 0.35,1.74±0.33,1.70±0.34,1.02±0.48,1.71±0.31 and 1.07±0.18,respectively,and the differences between the groups were statistically significant (F=3.791,P＜0.05).Among them,the LOVO cell viability of ERβ with estradiol group was lower than that of ERβ with ethanol group,accordingly,that of ERβ with MSH2 and estradiol group was lower than that of ERβ with MSH2 and ethanol group,that of ERβ with estradiol group was lower than that of empty plasmid with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t=3.158,3.075,3.648,3.253,all P＜0.05).DNA ladder formed from DNA fragments of apoptosis cells was seen in ERβ with estradiol group and ERβ with MSH2 and estradiol group.The apoptosis rate of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 7.86±0.19,7.87±0.39,8.39±1.02,9.05±1.54,7.54±0.99,19.77±2.35,7.76±1.32 and 19.30±1.75,respectively,and the differences between groups were statistically significant (F=45.436,P＜0.05).Among them,the apoptosis rate of ERβ with ethanol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and ethanol group was lower than that of ERβ with MSH2 and estradiol group,that of empty plasmid with estradiol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t =8.260,9.133,8.596,7.617,all P＜ 0.05).Conclusions Estrogen may promote colon cancer cell apoptosis through ERβ pathway.The process of apoptosis maybe related with caspase protein,MSH2 may not be involved in the regulation of this signal pathway.

Objective To investigate the effects of ciyelosporine-A(CsA).a calcinenrin(CAN)inhibitor,on electrophysiological propertiesof atria in canine tachycardia-induced model of AF.Methods Eighteen healthy adult mongrel canines weighing 17.0 to 23.2 kg(rangedfrom 2 to 4 years old)were randomized to 3 groups,Sham group(no pacemaker was implanted),atrial tachypacing group(ATP group)each group at baseline and after 8 weeks' tachypacing.Measurements included atrial effective refractory period(AERP),conductionvelocity(CV),wave length(WE),atrial fibrillation load and rate-adaptability. Results After 8 weeks' atrial tachypacing,ATP andCsA groups showed significant longer duration of the P wave,shorter AERP,decreased adaptation of AERE slower CV,shorter Wland longer AF duration compared to the shamg roup (all P＜0.05).AERP of the CsA group was longer than that of ATP group (P＜0.05),but there were no differences in rate-adaptability,CV,incidence of induced AF and AF duration between CsA group and ATP group.Conclusions Our results suggest that calcineurin pathway intervention by CsA have a positive effect on tachycardia-inducedelectrical remodeling of atria,but can not prevent or reverse AF.

Objective To explore whether ginsenoside Rg1 can attenuate ?-amyloid peptide 25-35-induced Tau hyperphosporylation in rat embryo cortical neurons by regulating the activity of GSK-3? and PP2A. Methods Primary cultures of cortical neurons were prepared from the embryonic day 18?2 in Sprague-Dawley rats. The experimental groups were designed as follows:1.Neurons culture (control group); 2. Neurons exposed to 20?mol/L A?_ 25-35 for 12 hours (A?-model group); 3.Neurons exposed to 20?mol/L A?_ 25-35 and 10 mmol/L lithium chloride (LiCl), a specific inhibitor of glycogen synthase kinase-3?(GSK-3?), for 12 hours (LiCl group); 4.Neurons exposed to 20?mol/L A?_ 25-35 for 12 hours in the presence of 24-hour pretreatment with ginsenoside Rg1 (Rg1 pretreatment group) . Western blotting and immunocytochemical staining were used to detect the levels of Tau phosphorylation,total Tau and GSK-3? in cortical neurons. Non-radioimmunoassay was introduced to detect the activity of protein phosphatase 2A (PP2A). Results In A?-model group, the levels of Tau protein phosphorylation in the sites of Ser 396 ,Ser 199/202 ,Thr 231 and total Tau were enhanced. Meanwhile, the expression of GSK-3? was also increased, but the activity of PP2A was unchanged. In LiCl group and Rg1 pretreatment group , the hyperposphorylations of Tau protein and total Tau and the expression of GSK-3? were markedly reduced compared to those of the A?-model group (P

Objective To investigate the expression and the effects of tissue inhibitor of metalloproteinases-1 (TIMP-1) on lungs of rats with sepsis.Methods Forty Sprague-Dawley (SD) rats were randomly divided into two groups,namely sham group (n =8) and sepsis model group (n =32).The rats of model group were modeled by cecal ligation and puncture (CLP),and were further divided into four subgroups as per the time after modeling,namely 6 h (n =8),12 h (n =8),24 h (n =8),48 h (n =8)subgroups.Blood and lung samples were taken 6 h,12 h,24 h and 48 h after modeling.The histological changes in lungs of the rats were observed under light microscope.Expressions of TIMP-1 mRNA,Bax mRNA and Bcl-2 mRNA in lungs were measured by RT-PCR.The immunohistochemistry was used to label the CD18 in lungs during different phases of sepsis.The data were processed by t test.Results Compared with sham group,the lung tissues of rats in model group were injured to a certain extent after CLP.The expression of TIMP-1 mRNA and the number of CD18 positive cells increased at the same time (P ＜ 0.01),and peaked 24 hours later (P ＜ 0.01).While the expression of Bax mRNA in model group decreased markedly 12-48 hours after modeling (P ＜ 0.01-0.05),and reached minimum 48 hours later (P ＜ 0.01).The expression of Bcl-2 mRNA in model group changed unnoticeable.The positive correlation between variations in number of CD18 positive cells and expression of TIMP-1 mRNA was found in model group (r =0.426,P ＜ 0.01).Conclusions The increase in expression of TIMP-1 mRNA in lungs is closely associated with the lung injury of sepsis.The mechanism of lung injury is likely attributed to the preservation of inflammatory cells from apoptosis,and the persistent inflammation response causes tissue damage,leading to organ dysfunction.

Objective To investigate the behavioral problems,intellectual ability and executive functioning characteristics as well as their correlations in children with borderline intellectual functioning (BIF) and attention deficit hyperactivity disorder (ADHD).Methods 60 children with BIF,60 children with ADHD and 60 typical developing children (TDC) participated this study.Parental report measures of behavioral problems were administrated using the Conners' Parent Symptom Questionnaire (PSQ) and Achenbach Children Behavior Checklist Scale (CBCL).All the participants performed the Chinese Revised Wechsler Intelligence Scale for Children (C-WISC) and a neuropsychological test named integrated visual and auditory continuous performance test (IVA-CPT).Comparison of cognitive profiles and correlation between psychological features were carried out to test the differences and commons among the groups.Results (1) Behavioral problems:the children with BIF had higher scores(3.68±2.85) in withdrawal factor than the children with ADHD (2.82±2.24) and TDC (1.03± 1.59).The children with ADHD had higher scores in conduct,impulsivity,aggressive and delinquent behavior than the children with BIF and TDC.The children with BIF/ADHD had higher scores in learning,anxiety,depression,somatic complaint and hyperactivity than the TDC.(2) Executive functioning:the children with BIF had more visual omission error (12.17± 10.48) than the children with ADHD (6.95±7.20) and TDC (2.33±2.48).The children with BIF and ADHD had lower auditory accuracy and higher visual reaction time,visual/auditory response variability,visual/auditory misrepresentation and auditory omission numbers than the TDC.These was no group difference in auditory reaction time.(3) Correlation analysis:intellectual ability was correlated with behavior problems and neurophysiological index such as conduct,learning,impulsivity,anxiety,depression factors,visual/auditory accuracy,visual reaction time and visual/auditory response variability.Conclusion BIF and ADHD are associated with poor executive functioning and increased behavior problems.The cognitive deficit may be a marker of vulnerability of poor academic and social outcomes in children with BIF and/or ADHD.

Objective:To investigate the characteristics of event-related potentials(ERPs)P300 in the development of attention cognitive function of school-age children.Methods:A total of 180 school-age children were divided into 3 groups according to their ages (7-8 years old group with 48 cases, 9-10 years old group with 44 cases and 11-12 years old group with 48 cases). All of the participants completed an Oddball task, and their behavioral data, P3a and P3b components of ERPs were analyzed by SPSS 21.0.Results:(1)The differences of hit numbers(49(47.25, 50), 50(49, 50), 50(50, 50) ), correct reaction time((533.37±56.94) ms, (486.91±61.12) ms, (411.55±51.97) ms), and Omission errors (2(1, 4), 2(1, 3), 1(0, 2) ) among the three groups were statistically significant( F/χ 2=20.635, 54.477, 13.169, all P<0.01). (2)There was a main effect of age( F=3.884, P=0.023) and an interaction effect between age and condition( F=3.314, P=0.038) on the amplitude of P3a, while the main effect of condition was not significant( F=0.111, P=0.740). The amplitude of P3a component in 11-12 years old group ((11.02±6.00)μV) was significantly larger than that of 7-8 years old group ((7.36±4.48)μV) and 9-10 years old group ((7.76±5.17)μV, both P<0.05). However, there was no significant interaction effect between age and condition on P3a latency, P3b amplitude and latency ( P>0.05). Conclusion:P3a may be a sensitive indicator of cognitive function in school-age children, and the age of 11-12 years old is a sensitive period for the development of children's attention orientation and selection ability, which may be related to the development of brain network.

Objective To investigate the levels of nutritional knowledge, attitude and dietary habits and its influential factors of the officers and soldiers in frontier forces to provide the theory basis for nutrition education.Methods Using self-designed nutritional knowledge-altitude-practice (KAP) questionnaires to survey 90 officers and soldiers who lived in plateau for at least 6 months. Questionnaires were handed out and handed over on the spot. According toMilitary Nutrition Survey and Evaluation Methods (GJB 1636A-2016), we measured the physical index of officers and soldiers, including height, weight and skin thickness of triceps. And we also evaluated body mass index (BMI) and upper arm muscle circumference.Results Nutrition knowledge, attitude and practice conditions of plateau forces were at the average, better and average levels respectively. Nutritional knowledge was significantly positively related to the practices (r=0.283,P=0.003), and was also significantly positively related to the age of the soldiers (r=0.228,P=0.04). Nutritional attitude was positively related to source of officers and soldiers (r=0.339,P=0.035), and nutritional practices were positively related to the upper arm muscle circumference (r=0.222,P=0.030). 65.6% of the officers and soldiers would like to know the knowledge about relationship between diet and disease. 25.6% of the officers and soldiers in the plateau were alcohol users.Conclusions Although the nutrition knowledge of plateau forces were poor, and the nutritional practices were influenced by nutritional knowledge, but the soldiers' attitude of changing unhealthy dietary habits was very positive. It is necessary to undertake the dietary nutrition education related to the high altitude nutrition among plateau forces to guild their nutritional practice scientifically.

Objective To investigate the dietary nutrition status of plateau frontier forces and to make recommendations on food and nutrients intake for plateau forces.Methods The dietary status of the plateau forces located at an altitude of 4600 m and 4030 m was investigated by weighing method.The dietary ration for soldiers, their daily dietary allowance and dietary balance index were used to evaluate their dietary patterns and nutrient intake.The concentrations of serum vitamin A and 25-OH vitamin D(25-OH VD) were detected by high performance liquid chromatography (HPLC).The arm muscle circumference and body fat rate of soldiers were evaluated by GJB 1636A-2016 and body composition related standard respectively.Results The intake of eggs, milk and plant oil of Unit A was sufficient, while the rest of the food intake did not reach the military standard;The intake of grain and plant oil of Unit B was sufficient,but the rest of the food intake was insufficient.The intake of protein,calcium,phosphonium,sodium,iron,selenium,iodine,copper,manganese,vitamin E and vitamin B3 of Unit A were adequate, but that of energy,potassium,zinc,magnesium,vitamin D,vitamin C,vitamin B1 and vitamin B2 was insufficient, and the intake of vitamin A,vitamin B6,vitamin B9 and vitamin B12 of Unit A was deficient.The intake of energy, protein,vitamin E,vitamin C,vitamin B1 and vitamin B3,and most of the minerals was adequate in Unit B, but vitamin B2 was insufficient.Furthermore, the intake of calcium,iodine,vitamin A,vitamin D and vitamin B was deficient.The proportion of nutrients which supply energy and the energy distribution of three meals in the two units were imperfect.The concentration of plasma vitamin A in both units was sufficient,but the concentration of plasma 25-OH VD was deficient.93.5% soldiers of unit A and 97.7% soldiers of unit B reached the standard of proper muscle circumference,80.6% soldiers of unit A and 70.5% soldiers of unit B had a low body fat rate.Conclusion The dietary structure of plateau border forces is not balanced.The intake of some foods and nutrients is insufficient so that nutritional education is badly needed to improve the dietary status of plateau forces.