AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.

Objective To evaluate the role of spinal sigma-1 receptors in the maintenance of bone cancer pain (BCP) in rats and the relationship with extracellular signal-regulated kinase (ERK).Methods Part Ⅰ Twenty-four female Sprague-Dawley rats,weighing 180-220 g,were randomized into 2 groups using a random number table:sham operation group (S group,n =4) and BCP group (n =20).BCP was induced by inoculating Walker 256 mammary gland carcinoma cells into the medullary cavity of the right tibia.Four rats were sacrificed on day 10 after inoculation in S group or on day 3,5,7,10 and 14 after inoculation in BCP group,and the L4-6 segments of the spinal cord were removed to measure the expression of sigma-1 receptors by Western blot.Part Ⅱ Forty female Sprague-Dawley rats,weighing 180-220 g,were randomized into 4 groups (n =10 each) using a random number table:sham operation group (S group),sigma-1 receptor inhibitor BD1047 group (BD group),BCP group,and BCP + BD1047 group (BCP + BD group).On day 10 to 14 after inoculation,normal saline 20 μl was injected intrathecally once a day in S and BCP groups,or BD1047 120 nmol/20μl was injected intrathecally once a day in BD and BCP + BD groups.Mechanical paw withdrawal threshold (MWT) to yon Frey filament stimulation was measured one day before inoculation,on day 3,5 and 7 after inoculation,and on day 10,12 and 14 after administration.After measurement of MWT on day 14 after inoculation,the rats were sacrificed and the L4-6 segments of the spinal cord were removed to determine the expression of phosphorylated ERK (p-ERK) by Western blot.Results Part Ⅰ Compared with group S,the expression of sigma-1 was significantly up-regulated and peaked on day 10 after operation in group BCP.Part Ⅱ Compared with S group,no significant changes were found in MWT and p-ERK expression at each time point in BD group,and MWT was decreased and p-ERK expression was up-regulated in BCP and BCP + BD groups.Compared with group BCP,after intrathecal injection of BD1047,MWT was significantly increased and the expression of p-ERK was down-regulated in BCP + BD group.Conclusion Spinal sigma-1 receptors are involved in the maintenance of BCP in rats possibly through promoting phosphorylation of ERK.

[ ABSTRACT] AIM:To investigate the cardiac AMP-activated protein kinase ( AMPK) activity and the effects of AMPK activator on cardiac structure and function in the mice with different β-adrenoceptor (β-AR) stimulation patterns . METHODS:Male BALB/c mice were subcutaneously injected with AMPK activator ( AICAR, 250 mg· kg -1 · d-1 ) or saline, and infused with β-AR agonist isoproterenol (ISO, 5 mg· kg-1· d-1) for 14 d.The cardiac functions were evalu-ated by echocardiography or hemodynamic method , and the hearts were harvested after infusion cessation immediately or 3 d later.Phosphorylated AMPK ( p-AMPK) was measured by Western blot .RESULTS:Sustained ISO infusion increased p-AMPK level.AICAR did not further increase p-AMPK but attenuated ISO-induced increase in heart weight .Sustained ISO infusion increased cardiac systolic function as indicated by left ventricular fractional shortening ( FS) and maximum rate of pressure rise (+dp/dtmax).The cardiac systolic function was not further increased by AICAR .The cardiac diastolic func-tion as indicated by left ventricular end-diastolic pressure (LVEDP) was not different in each group .In contrast, cardiac p-AMPK level was similar between the control mice and the mice with sustained ISO infusion and ceased infusion for 3 d. In this model, AICAR improved the cardiac systolic and diastolic functions , which were impaired by ISO.Moreover, the increased pattern of p-AMPK level was similar with that of heart rate upon ISO stimulation .CONCLUSION: Sustained ISO infusion increases p-AMPK.After ISO infusion cessation for 3 d, p-AMPK is decreased to the basal level .β-AR-in-duced inotropic effects should be avoided to investigate the cardioprotective role of AMPK activation in the β-AR stimulation models.

Objective To describe the understanding level of disease of children with leukemia and their ways of obtaining disease information in order to help nurses and parents to select appropriate content and manners to communicate with children about disease information.Methods In-depth interviews were conducted with 25 children' parents using a descriptive qualitative research method,and the data were analyzed using content analysis.Results Children during remission stage of leukemia had different understanding levels of their disease.Ways of children with leukemia to obtain disease information was correlated with their mental maturity.Conclusions Disease information should be told according to children's age,disease course and level of thinking,and health professionals and parents could provide appropilate ways of obtaining information for children on basis of their mental maturity.

group Nor(P0.05).Conclusions The changes of ambulatory pulse pressure can reflect the degree of EH.The obvious increase of PP and decrease of DBP maybe are the exclusive features of ABPM in EH patients with target organs damage.

Objective To investigate the current situation of maternal self- efficacy of breastfeeding and explore its influencing factors in Zhengzhou. Methods Using the general condition questionnaires, breast feeding self-efficacy scale, perceived social support scale and Edinburgh postnatal depression scale to assess the situation of 180 puerperas by the convenience sampling in one obstetric hospital of Zhengzhou. Results The total score of maternal breast feeding self-efficacy was(114.04 ± 21.57)points.The frequency of delivery, the way of delivery and feeding ways, the average income of family were effected with breastfeeding self-efficacy. The total score of social support was(68.87 ± 10.43) points, the total score of puerperas depression was(7.61 ± 4.25) points.The social support score had positive correlation with breastfeeding self-efficacy(r=0.423, P<0.01). Puerperas depression had negative correlation with breastfeeding self- efficacy(r=- 0.342, P<0.01). Conclusions The maternal breastfeeding level in Zhengzhou was at a lower level. The level of maternal breastfeeding self-efficacy scale was impacted by maternal social support degree, the degree of depression. Measures should be taken to enhance maternal social support degree, reduce the degree of depression, in order to improve the level of maternal breastfeeding self-efficacy.

AIM: To determine the effect of suction duration on thickness and diameter of corneal flap created by microkeratome in porcine eyes in laser in situ kerato-mileusis (LASIK).METHODS: Sixty porcine eyes were randomly assigned to three groups according to different suction durations: group 1 (10 seconds), group 2 (20 seconds), and group 3 (30 seconds). A Moria M2 microkeratome (Moria, France) with a 160μm head was used to create a corneal flap. Corneal flap thickness was measured by automated ultrasonic pachymetry, and the flap diameter was measured by a vernier caliper.RESULTS: The flap thickness of group 1, group 2 and group 3 was 146.05±13.46μm, 157.35±18.95μm and 169.25±21.02μm, respectively. There was a statistically significant difference among three groups (P=0.001). The mean flap diameter in groups 1, 2 and 3 was 8.63±0.19mm, 8.89±0.24mm and 9.06±0.18mm, respectively. A statisti-cally significant difference was found among groups (P<0.01).CONCLUSION: In LASIK in porcine eyes, an increase in suction duration resulted in a thicker and greater flap.

The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.

Objective To investigate the risk factors resulting in the perioperative liver failure and death for the HBV-associated hepatocellular carcinoma (HCC) patients with or without cirrhosis.Methods The method of retrospective case-control study was performed.The clinicopathological data of 1 083 HCC patients with positive HBsAg who received curative liver resection at the Southwest Hospital from January 2008 to December 2012 were collected.According to the absence or presence of cirrhosis,the HCC patients with positive HBsAg were divided into the 2 groups,including the cirrhosis group (633 patients) and the non-cirrhosis group (450patients).The intraoperative conditions (operation time,volume of intraoperative blood loss,rate of blood transfusion,rate of pringle maneuver) and postoperative conditions (incidence of perioperative complications,duration of postoperative hospital stay,perioperative mortality) of HCC patients were observed.The gender,age,alanine transaminase (ALT),aspartate transaminase (AST),albumin (Alb),total bilirubin (TBil),platelet (PLT),Child-Pugh classification,operation time,volume of intraoperative blood loss,blood transfusion,pringle maneuver,extent of liver resection,number of tumors,tumor diameter,tumor thrombus and liver cirrhosis were enrolled and prognostic factors resulting in perioperative liver failure and death for the HCC patients were explored.Measurement data with skewed distribution were presented as M (range) and comparison between the 2 groups was analyzed using Mann-Whitney U test.Count data were presented as counts (percentage) and comparison between the 2 groups was analyzed using chi-square test or Fisher exact probability.Univariate analysis was performed by chi-square test and multivariate analysis was performed by Logistic regression model (forward).Results (1) The intraoperative conditions:the volume of intraoperative blood loss were 500 mL (range,30-7 000 mL) in the cirrhosis group and 400 mL (range,50-8 000 mL) in the non-cirrhosis group,with a statistically significant difference between the 2 groups (Z =-2.209,P ＜ 0.05).The operation time,rate of blood transfusion and rate of pringle maneuver were 250 minutes (range,82-715 minutes),29.86％ (189/633),62.24％ (394/633) in the cirrhosis group and 242 minutes (range,85-738 minutes),27.11％ (122/450),66.67％ (300/450) in the non-cirrhosis group,respectively,with no statistical differences between the 2 groups (Z =-1.212,x2 =0.969,2.236,P ＞0.05).(2) The postoperative conditions:the incidence of perioperative complications was 30.49％(193/633) in the cirrhosis group and 21.11％ (95/450) in the non-cirrhosis group,with a statistically significant difference between the 2 groups (x2 =11.851,P ＜ 0.05).The incidence of lung infection,abdominal infection and liver failure were 6.48％ (41/633),2.69％ (17/633),5.53％ (35/633) in the cirrhosis group and 3.56％ (16/450),0.89％ (4/450),1.33％ (6/450) in the non-cirrhosis group,respectively,with statistically significant differences between the 2 groups (x2 =4.502,4.465,12.713,P ＜ 0.05).The duration of postoperative hospital stay was 15 days (range,0-70 days) in the cirrhosis group and 14 days (range,0-71 days) in the non-cirrhosis group,with a statistically significant difference between the 2 groups (Z =-3.448,P ＜ 0.05).The perioperative mortality was 5.85％ (37/633) in the cirrhosis group and 2.44％ (11/450) in the non-cirrhosis group,with a statistically significant difference between the 2 groups (x2=7.181,P ＜ 0.05).(3)Results of risk factors affecting perioperative liver failure:①results of univariate analysis showed that age,AST,Alb,Child-Pugh classification,operation time,volume of intraoperative blood loss,blood transfusion,extent of liver resection,tumor diameter,liver cirrhosis with positive HBsAg were associated with perioperative liver failure in HCC patients (x2=5.013,7.979,8.855,16.968,14.148,9.764,18.511,11.749,5.534,12.713,P＜0.05);age,AST,Alb,Child-Pugh classification,operation time,blood transfusion,extent of liver resection and tumor diameter were associated with perioperative liver failure in the cirrhosis group (x2=5.877,5.380,11.087,13.672,8.849,13.170,12.418,5.805,P ＜ 0.05);volume of intraoperative blood loss was associated with perioperative liver failure in the non-cirrhosis group (P ＜ 0.05).②Results of multivariate analysis showed that age≥60 years,Child-Pugh class B,operation time ＞ 360 minutes,blood transfusion,extent of liver resection ≥3 segments and liver cirrhosis were independent risk factors affecting perioperative liver failure in HCC patients with positive HBsAg [OR =2.285,2.716,2.315,2.159,2.459,4.322;95％ confidence interval (CI):1.081-4.831,1.100-6.706,1.064-5.038,1.068-4.362,1.264-9.786,1.763-10.598,P＜0.05];Alb ＜38 g/L,Child-Pugh class B,blood transfusion and extent of liver resection ≥ 3 segments were independent risk factors affecting perioperative liver failure in the cirrhosis group (OR =2.231,2.857,2.186,2.927,95％ CI:1.038-4.795,1.095-7.451,1.045-4.576,1.426-6.008,P ＜ 0.05);volume of intraoperative blood loss ＞ 1 200 mL was an independent risk factor affecting perioperative liver failure in the non-cirrhosis group (OR =15.077,95％CI:2.695-84.353,P ＜ 0.05).(4) Risk factors affecting perioperative death:①results of univariate analysis showed that gender,Alb,TBil,Child-Pugh classification,blood transfusion,extent of liver resection,tumor diameter,tumor thrombus and liver cirrhosis were associated with perioperative death in HCC patients with positive H BsAg (x2=4.462,8.783,4.212,4.869,7.189,11.745,6.837,4.323,7.181,P ＜0.05);Alb,extent of liver resection and tumor diameter were associated with perioperative death in the cirrhosis group (x2=12.173,12.793,10.981,P ＜ 0.05);blood transfusion and tumor thrombus were associated with perioperative death in the non-cirrhosis group (x2 =5.836,6.417,P ＜ 0.05).② Results of multivariate analysis showed that Alb ＜38 g/L,extent of liver resection ≥ 3 segments and liver cirrhosis were independent risk factors affecting perioperative death in HCC patients with positive HBsAg (OR =2.560,2.657,2.567,95％ CI:1.382-4.742,1.471-4.800,1.283-5.134,P ＜ 0.05);Alb ＜ 38 g/L,extent of liver resection ≥ 3 segments and tumor diameter≥5 cm were independent risk factors affecting perioperative death in the cirrhosis group (OR =3.003,2.533,3.060,95％ CI:1.495-6.034,1.251-5.128,1.135-8.251,P＜0.05);blood transfusion and tumor thrombus were independent risk factors affecting perioperative death in the non-cirrhosis group (OR =3.755,4.036,95％ CI:1.047-13.467,1.126-14.469,P ＜ 0.05).Conclusions Liver cirrhosis is an independent risk factor for perioperative liver failure and death in HCC patients with positive HBsAg.The risk of perioperative liver failure and death in HCC patients with cirrhosis is significantly higher than that in HCC patients without cirrhosis,and there is a difference in the risk factors for perioperative liver failure and death.

Objective To study the correlation between serum homocysteine ( Hcy ) level and C677T polymorphism of methylenetetrahydrofolate reductase ( MTHFR ) gene C677T polymorphism ( rs1801133) in patients with cerebral infarction, and feature of rs1801133 polymorphism and serum Hcy level in cerebral infarction patients with or without diabetes mellitus.Methods Case-control study.Five hundred and fifty six patients with cerebral infarction admitted to China-Japan Friendship Hospital from January 2014 to January 2015 were included as the case group while 275 subjects from medical examination center without cerebral infarction and diabetes mellitus matched with the case group.MTHFR C677T polymorphism was determined by pyrosequencing and serum Hcy was determined by circulating enzymatic.Chi-square test was used to analyze the distribution of genotype in different group; ANVOA was used to analyze the Hcy level with different genotype in patients with cerebral infarction, and LSD-t was used to pairwise comparison.Results Among the 556 patients with cerebral infarction ,TT genotype were 202 cases (36.33%), CT genotype were 257 cases(46.22%), CC genotype were 97 cases(17.45%).The T allele 44%, higher than the control group T allele frequencies 46.91%(χ2 =23.385,P<0.001).The level of TT genotype serum Hcy level (21.31 ±17.31) μmol/L were higher than CT genotype (14.88 ±7.71) μmol/L(P<0.001)and CC genotype(14.48 ±7.78) μmol/L(P<0.001).There is no significant statistics different in TT genotype frequency between Cerebral infarction patients with diabetes mellitus(36.77%) and without diabetes mellitus(36.44%) (χ2 =0.031,P>0.05), while the level of serum Hcy in Cerebral infarction patients with diabetes mellitus ( 18.16 ±12.90 )μmol/L is lower than Cerebral infarction patients without diabetes mellitus(23.47 ±19.53) μmol/L in TT genotype( F=4.652, P<0.05).Conclusions MTHFR TT genotype was related to serum hyperhomocysteine, and maybe save as the risk of cerebral infarction.The Hcy level in TT genotype cerebral infarction patients with DM is lower than the same genotype patients without DM.(Chin J Lab Med, 2016, 39:205-209 )