Objective To investigate the relationship between coagulation function and pediatric critical illness scores (PCIS) in children with severe pneumonia.Methods The PCIS were collected in 152 children with severe pneumonia (pneumonia group) admitted in our pediatric intensive care unit from Jan 2010 to Jul 2011,and 20 healthy children in the same period were selected as the healthy control group.The coagulation indicators of children in both groups were detected and the relationship between coagulation markers and severity of pneumonia was analyzed.Results There were significant differences in platelet count,fibrinogen,D-Dimer,soluble P-selectin between pneumonia group and healthy control goup [(185.74 ±116.26) × 109/L vs (287.10 ±90.01) × 109/L,(3.51 ±0.50) g/L vs (3.15 ±0.15) g/L,(1.39 ±2.18) μg/ml vs (0.36 ± 0.07) μ g/ml,(110.07 ± 83.47) ng/ml vs (33.74 ± 9.47) ng/ml,P ＜ 0.05].There were positive correlation between soluble P-selectin,D-Dimer and severity of disease and negative correlation between platelet count and severity of disease in children with severe pneumonia.Regression equation:y =1.154 +0.003 × soluble P-selectin + 0.089 × D-Dimer-0.001 × platelet count (P ＜ 0.05).As the children's critical condition getting worse,soluble P-selectin and D-Dimer levels increased (P ＜ 0.05).Hatelet count showed no significant difference between critical group and extremely critical group,which was significantly lower than that in non-critical group (P ＜ 0.05).Conclusion Soluble P-selectin,D-Dimer,and platelet count are associated with the severity of pneumonia.The children with severe pneumonia are easy to have coagulation disturbance.

Objective To observe the effect of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on cholesterol gallstones formation in C57BL/6 mice with diet-induced cholesterol gallstone,and then explore the potential mechanism.Methods Fifty C57BL/6 mice were randomly divided into 5 groups (10 mice in each group),referring to control group,experimental group,experimental plus DHA group,experimental plus EPA group,as well as experimental plus DHA and EPA group.The mice in control group were fed with regular diet,and the rest of the mice with lithogenic diet (LD).Subsequent to feeding the mice with separate diets for two weeks,EPA and/or DHA (70 mg · kg-1 · d-1) were orally administered for eight weeks,while the LD feeding was continued during this period.After a total of 10 weeks,the mice were dissected to observe the gallstone formation.The levels of serum lipids,total cholesterol (TC) and phospholipids (PL) in bile,and TC in the liver were tested,and the protein expression of HMGCR,SRBI,ABCG5/ABCG8,CYP7A1 and ABCB11genes in the liver of mice was measured.Results Compared with the experimental group,the experimental plus EPA group had significantly lower TC in liver (0.033 ±0.008 mmolo/g) and bile (1.807 ±0.381 mmolo/L),and lower relative protein expression levels of HMGCR (0.545±0.098),ABCG5 (0.418±0.089) and ABCG8 (0.501 ±0.151)in liver (P＜ 0.05).The contents of TC in liver and bile,and the protein expression of HMGCR,ABCG5andABCG8 in liver were 0.048 ± 0.006 mmol/g and 2.662 ± 0.339 mmolo/L,and 1.011 ± 0.213,1.037 ± 0.276 and 1.266 ±0.312,respectively.No significant differences were observed between experimental plus DHA group and experimental group (P ＞ 0.05).Conclusions EPA could prevent the cholesterol gallstone formation in mice by decreasing the expression of HMGCR and ABCG5/8 genes in liver,therefore reducing cholesterol synthesis and blocking cholesterol transport from liver to bile as well as diminishing cholesterol content in the bile.However,the inhibition effect of DHA on cholesterol gallstone formation was not obvious.

Objective To explore the effect of exogenous recombinant human erythropoietin (rhEPO) on neuronal apoptosis in neonatal rats after hyperoxia brain injury.Methods Thirty neonatal Wistar rats were randomly divided into 3 groups by random number table method:rhEPO treatment + 800 mL/L hyperoxia group (group A),9 g/L saline +800 mL/L hyperoxia group (group B),9 g/L saline + air group (group C).Group A was given subcutaneous injection of rhEPO 1 000 IU/kg for 5 days.Group B and group C received the same dose of 9 g/L saline.Group A and group B were continuously exposed to atmospheric pressure hyperoxia model cabin to maintain the oxygen concentration in the container (800 ± 30) mL/L for 5 days.During the course of the experiment,the general situation and weight changes in rats were observed.After 5 d,all rats were sacrificed and brain tissues were taken.Neuronal apoptosis in hippocampal structural region of the newborn rats was observed by terminal deoxynucleotidyl transferase dUTP nick and labeling(TUNEL) staining.Immunohistochemical method was used to detect the expression of 5-lipoxygenase in hippocampal structural region of newborn rats.Results The weight gain and brain weight of group B were lower than those of group C,the weight gain and brain weight of group A were higher than those of group B,and the differences were statistically significant(F =11.179,8.140,all P ＜ 0.05).In group A and group B were found that the neuronal nucleus of the hippocampal neurons was partially contracted,deeply dyed,and the neuronal arrangement was loose,even with local neuron deletions and focal necrosis,but in group A neuron density was higher with less necrosis than that in group B.The neuronal cells in hippocampal structural region were neat and intact in group C.The number of TUNEL positive cells in hippocampal structural region of group B[(6.20 ± 1.93) number/high power field] was significantly higher than that in group C [(1.80 ± 0.79) number/high power field],the number of TUNEL positive cells in hippocampal structural region of group A [(4.20 ± 1.32) number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =23.912,P ＜ 0.05).The number of 5-lipoxygenase positive cells in group B [(6.90 ± 1.29) number/high power field] was significantly higher than that in group C [(1.00 ± 0.67) number/high power field],the number of 5-lipoxygenase positive cells in group A [(5.60 ± 0.97)number/high power field] was significantly lower than that in group B,and the difference was statistically significant (F =95.044,P ＜ 0.05).Conclusion rhEPO has a protective effect on neonatal rats with hyperoxia brain injury,and alleviates brain cell apoptosis caused by hyperoxia brain injury,which may interfere with the 5-lipoxygenase pathway.

Objective To investigate the effect and mechanism of high concentration glucose on cholesterol absorption of human colon cancer epithelial Caco-2 cells.Methods The experimental study was used.(1) CCK-8 detected cell proliferation:the proliferation rate changes of Caco-2 cells were detected by CCK-8 when different concentrations (12.5,100.0,300.0,700.0,1 000.0,1 388.0 mmol/L) of glucose solution effects on Caco-2 cells in order to ensure the half hindering concentration of glucose concentration on Caco-2 cells.(2)Cholesterol absorption of Caco-2 cells was detected:Caco-2 cells were divided into the cholesterol group,cholesterol plus ezetimibe (cholesterol inhibitor) group and blank control group.Cholesterol group:100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Cholesterol plus ezetimibe group:100 μmol/L ezetimibe,100 μmol/L cholesterol solution and different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added.Blank control group:DMEM culture medium and corresponding concentrations of DMSO were added.The cholesterol absorption amounts of Caco-2 cells were measured.(3) The relative expressions of ATP binding cassette G8 (ABCG8),ATP binding cassette G5 (ABCG5),Nickman-Pick CI Like 1 (NPC1L1) and scavenger receptor class B type Ⅰ (SR-B Ⅰ) were examined by Western blot in the different groups.Caco cells were divided into the glucose group,glucose plus ezetimibe group and control group.The different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution were added into the glucose group,different concentrations (5.0,25.0,50.0 mmol/L) of glucose solution and 100 μmol/L ezetimibe were added into the glucose plus ezetimibe group,and 100 μmol/L ezetimibe were added into the control group.The relative expressions of ABCG8,ABCG5,NPC1L1 and SR-B Ⅰ were detected by Western blot.Measurement data were presented as (x) ±s,repeated measure variance analysis was used to perform variation trend test,and t test was utilized to conduct comparisons among groups.Results (1) CCK-8 results showed:proliferation rates of Caco-2 cells with the glucose solution concentration of 12.5,100.0,300.0,700.0,1 000.0 and 1 388.0 mmol/L were 1.380 ±0.043,1.238 ±0.072,0.736 ±0.035,0.336 ±0.021,0.316 ±0.020 and 0.288 ±0.010,respectively,with a statistically significant difference in the proliferation rates (F =11.019,P ＜ 0.05).The half hindering concentration of glucose solution on Caco-2 cells was 283.54 mmol/L.(2)Cholesterol absorption of Caco-2 cells:① the cholesterol absorption amounts of Caco-2 cells with the glucose solution concentration of 5.0,25.0 and 50.0 mmol/L were 0.282 ± 0.042,0.380 ± 0.063,0.390 ± 0.060 in the cholesterol group and 0.042 ± 0.012,0.197 ± 0.015,0.277 ± 0.029 in the cholesterol plus ezetimibe group,respectively,showing a statistically significant difference between the 2 groups (F =55.566,P ＜ 0.05).②There was a statistically significant difference in cholesterol absorption amounts of Caco-2 cells with different glucose solution concentration in the cholesterol group (F =79.117,P ＜ 0.05).The cholesterol absorption amounts of Caco-2 cells with the glucose solution concentrations of 5.0 mmol/L was lower than that with the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L,respectively (t =11.207,11.532,P ＜0.05).There was no statistically significant difference in the cholesterol absorption amounts of Caco-2 cells between the glucose solution concentrations of 25.0 mmol/L and 50.0 mmol/L (t =12.389,P ＞ 0.05).③There were statistically significant differences in cholesterol absorption amounts of Caco-2 cells with the glucose concentration of 5.0 mmol/L and 25.0 mmol/L between cholesterol group and cholesterol plus ezetimibe group (t =10.908,10.644,P ＜ 0.05).(3) The results of Western blot showed:① the relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L were respectively 0.277 ±0.019,0.558 ±0.015,0.576 ±0.003 in the glucose group and 0.057 ±0.002,0.054 ±0.005,0.077 ±0.005 in the glucose plus ezetimibe group,showing a statistically significant difference (F =482.207,P ＜0.05).② The relative expression of NPC1L1 protein of Caco-2 cells with the different concentration of glucose solution in the glucose group were compared,with a statistically significant difference (F =8.112,P ＜ 0.05).There was a statistically significant difference in the relative expression of NPC1L1 protein in Caco-2 cells with the different concentration of glucose solution in the glucose plus ezetimibe group (F =11.708,P ＜ 0.05).③ The relative expression of NPC1L1 protein in Caco-2 cells with the glucose solution concentrations of 5.0,25.0 and 50.0 mmol/L in the glucose group was statistically different from that in the glucose plus ezetimibe group (t =8.112,11.708,13.920,P ＜ 0.05).Conclusion High concentration glucose solution could promote the reabsorption of cholesterol through increasing NPC1L1 protein expression in Caco-2 cells,and increase the risk of suffering from cholelithiasis in diabetes patients.

Objective To explore the efficacy and safety of high-dose second-generation fat emulsion usage on the very low birth weight premature infants.Methods A total of 88 premature infants with very low birth weight (VLBW) in Neonatal Intensive Care Unit (NICU)of Fujian Provincial Maternity and Children Hospital,Affiliated Hospital of Fujian Medical University from December 2013 to December 2014 were randomly divided into experimental group and control group,with 44 cases in each group according to the table of random number.The experimental group received intravenous nutrition with 200 g/L second-generation fat emulsion within 24 hours after birth,the initial dose was 2.0 g/(kg · d) with an increase of (0.5-1.0) g/(kg · d) daily,the maximum dose was 3.5 g/(kg · d);the control group received intravenous nutrition with 200 g/L second-generation fat emulsion 24 hours later after birth,the initial dose was 0.5 g/(kg · d) with an increase of 0.5 g/(kg · d) daily,the maximum dose was 3.5 g/(kg · d).The other intravenous nutrition methods were same.The general conditions at birth,blood biochemical parameters,growth parameters and complications were compared between the 2 groups.Results The mean value of intravenous nutrition duration,length of stay,the glucose infusion rates of postnatal days 6 and 7,the serum triglyceride levels of postnatal days 7,chest circumference of the fourth weeks,the incidence of the low triiodothyronine(T3) syndrome and parenteral nutrition associated cholestasis(PNAC) were (22.27 ± 7.17) d,(37.75 ± 12.28) d,(8.10 ± 0.92) mg/(kg · min),(8.49 ± 1.06) mg/(kg · min),0.18(0.03-0.59) mmol/L and (27.21 ± 1.62) cm in the experimental group respectively,but (27.36 ± 11.37) d,(44.36 ± 16.45) d,(7.98 ±0.79) mg/(kg · min),(8.22 ±0.76) mg/(kg · min),0.28 (0.07-0.99) mmol/L and (26.56 ± 0.96) cm in the control group,respectively,and the differences were statistically significant between the 2 groups (t =2.512,5.403,4.314,9.705,696.500,6.668,all P ＜ 0.05).The incidence of the T3 syndrome and parenteral nutrition associated cholestasis (PNAC) in the experimental group was 25.0％ (11/44 cases) and 0(0/44 cases),respectively,which were significantly lower than those in the control group[81.8％ (36/44 cases)and 9.1％ (4/44 cases)],and the differences were statistically significant between the 2 groups (x2 =28.542,5.736,all P ＜ 0.05).The 2 groups had no significant difference in the incidence rates of other complications such as necrotizing enterocolitis,infection,retinopathy of prematurity,bronchopulmonary dysplasia,and the duration of oxygen therapy and mechanical ventilation(all P ＞ 0.05).Conclusions The high-dose second-generation fat emulsion usage [the initial dose 2.0 g/(kg · d)] in VLBW infants is safe and well tolerated.Advisable parenteral nutrition support strategy can promote growth of VLBW infants,shorten the intravenous nutrition duration and length of stay,reduce the incidence of the low T3 syndrome and PNAC,which has no influence on the incidence rates of other complications.

In this study, the mobilomes of nine teleost species were annotated by bioinformatics methods. Both of the mobilome size and constitute displayed a significant difference in 9 species of teleost fishes. The species of mobilome content ranking from high to low were zebrafish, medaka, tilapia, coelacanth, platyfish, cod, stickleback, tetradon and fugu. Mobilome content and genome size were positively correlated. The DNA transposons displayed higher diversity and larger variation in teleost (0.50% to 38.37%), was a major determinant of differences in teleost mobilomes, and hAT and Tc/Mariner superfamily were the major DNA transposons in teleost. RNA transposons also exhibited high diversity in teleost, LINE transposons accounted for 0.53% to 5.75% teleost genomic sequences, and 14 superfamilies were detected. L1, L2, RTE and Rex retrotransposons obtained significant amplification. While LTR displayed low amplification in most teleost with less than 2% of genome coverages, except in zebrafish and stickleback, where LTR reachs 5.58% and 2.51% of genome coverages respectively. And 6 LTR superfamilies (Copia, DIRS, ERV, Gypsy, Ngaro and Pao) were detected in the teleost, and Gypsy exhibits obvious amplication among them. While the SINE represents the weakest ampification types in teleost, only within zebrafish and coelacanth, it represents 3.28% and 5.64% of genome coverages, in the other 7 teleost, it occupies less than 1% of genomes, and tRNA, 5S and MIR families of SINE have a certain degree of amplification in some teleosts. This study shows that the teleost display high diversity and large variation of mobilome, there is a strong correlation with the size variations of genomes and mobilome contents in teleost, mobilome is an important factor in determining the teleost genome size.

Estrogen receptor (esr) mediates the effects of estrogen on the expression of related genes, thereby regulating the growth and reproduction of mammals. To investigate the effect of retrotransposon insertion polymorphism (RIP) of the porcine esr gene on porcine growth performance, retrotransposon insertion polymorphism of the esr gene were predicted by comparative genomics and bioinformatics, and PCR was used to verify the insertion polymorphisms in different porcine breeds. Finally, the correlation analysis between the genotypes and performance of Large White pigs was conducted. The results showed that four retrotransposon polymorphic sites were identified in the esr1 and esr2 genes, which are esr1-SINE- RIP1 located in intron 2 of the esr1 gene, esr1-LINE-RIP2 and RIP3-esr1- SINE located in intron 5 of the gene, and esr2-LINE-RIP located in intron 1 of the esr2 gene, respectively. Among them, insertion of a 287 bp of SINE into intron 2 of the esr1 gene significantly affected (P<0.05) the live back fat thickness and 100 kg body weight back fat thickness of Large White pigs. Moreover, the live back fat thickness and back fat thickness at 100 kg body weight of homozygous with insertion (SINE+/+) was significantly greater than that of heterozygous with insertion (SINE+/-) and homozygous without insertion (SINE-/-). Therefore, esr1-SINE-RIP1 could be used as a molecular marker to assist the selection of deposition traits in Large White pigs.
Animals ; Genotype ; Introns/genetics* ; Phenotype ; Polymorphism, Genetic/genetics* ; Retroelements/genetics* ; Swine/genetics*

With the completion of large-scale genome sequencing of human beings and other organisms, understanding the expression of control elements on the genome has become an important research task in the post-genome era. The enhancer trapping technology is an effective method for identifying enhancer elements in the genome and understanding its mechanism for gene expression regulation. In this study, we selected the stable enhancer trapping line TK4 (head and trunk specific GFP expression), which is generated with the mediation of Tol2 transposon system, and analyzed the trapped enhancers with the techniques of Splinkerette PCR (sp-PCR), in situ hybridization and comparative genomics. We crossed F1 individuals of TK4 line with wild-type zebrafish, collected fertilized eggs, and then detected the expression pattern of green fluorescent protein reporter gene by fluorescence microscopy at six different developmental stages, 6 hpf (hour post fertilization), 24 hpf, 48 hpf, 3 dpf (day post fertilization), 4 dpf and 5 dpf . The zebrafish genome flank sequence near the insertion site of Tol2 transposon was cloned by sp-PCR, and the results revealed that the insertion located at the position 27749253 of chromosome 23, and the transgene inserted reversely inside the intron 1 of rps26 gene. Within the 100 kb region of the insertion site, totally, seven genes including arf3a, wnt10b, wnt1, rps26, IKZF4, dnajc22 and lmbr1l were identified. Comparative genomic analysis by VISTA program revealed that there were two potential enhancer elements in the downstream of rps26 gene, which were conserved non-coding sequence (CNS) 1 and CNS2. The results of in situ hybridization showed that two transcripts of rps26 gene were maternal expression, the expression of rps26-201 in zygote was earlier than that of rps26-001, and the GFP signal of TK4 line zebrafish was not detectable before 6hpf, the expression patterns of rps26 and GFP at the late stages display similarity, and also represent differences, which suggested that the expression of rps26 and GFP may be controlled by the same enhancer, and also by the different enhancer, and two potential enhancers (CNS1 and CNS2) may play a differential regulation roles on the spatial and temporal expression of nearby genes (including rps26). In this study, we successfully obtained two potential enhancers near rps26 gene for the first time, which laid a foundation for further study of the regulation mechanism between these two enhancers and nearby genes in the genome, and the combination technique used in this study also provides a reference for enhancer analysis.