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Objective: To investigate the effect of targeted carboxylesterase 1f (Ces1f) gene knockdown on the polarization activity of Kupffer cells (KC) induced by lipopolysaccharide/D-galactosamine (LPS/D-GalN) in mice with acute liver failure. Methods: The complex siRNA-EndoPorter formed by combining the small RNA (siRNA) carrying the Ces1f-targeting interference sequence and the polypeptide transport carrier (Endoporter) was wrapped in β-1, 3-D glucan shell to form complex particles (GeRPs). Thirty male C57BL/6 mice were randomly divided into a normal control group, a model group (LPS/D-GalN), a pretreatment group (GeRPs), a pretreatment model group (GeRPs+LPS/D-GalN), and an empty vector group (EndoPorter). Real-time fluorescent quantitative PCR and western blot were used to detect Ces1f mRNA and protein expression levels in the liver tissues of each mouse group. Real-time PCR was used to detect the expression levels of KC M1 polarization phenotypic differentiation cluster 86(CD86) mRNA and KC M2 polarization phenotypic differentiation cluster 163 (CD163) mRNA in each group. Immunofluorescence double staining technique was used to detect the expression of Ces1f protein and M1/M2 polarization phenotype CD86/CD163 protein in KC. Hematoxylin-eosin staining was used to observe the pathological damage to liver tissue. A one-way analysis of variance was used to compare the means among multiple groups, or an independent sample nonparametric rank sum test was used when the variances were uneven. Results: The relative expression levels of Ces1f mRNA/protein in liver tissue of the normal control group, model group, pretreatment group, and pretreatment model group were 1.00 ± 0.00, 0.80 ± 0.03/0.80 ± 0.14, 0.56 ± 0.08/0.52 ± 0.13, and 0.26 ± 0.05/0.29 ± 0.13, respectively, and the differences among the groups were statistically significant (F = 9.171/3.957, 20.740/9.315, 34.530/13.830, P < 0.01). The percentages of Ces1f-positive Kupffer cells in the normal control group, model group, pretreatment group, and pretreatment model group were 91.42%, ± 3.79%, 73.85% ± 7.03%, 48.70% ± 5.30%, and 25.68% ± 4.55%, respectively, and the differences between the groups were statistically significant (F = 6.333, 15.400, 23.700, P < 0.01). The relative expression levels of CD86 mRNA in the normal control group, model group, and pretreatment model group were 1.00 ± 0.00, 2.01 ± 0.04, and 4.17 ± 0.14, respectively, and the differences between the groups were statistically significant (F = 33.800, 106.500, P < 0.01). The relative expression levels of CD163 mRNA in the normal control group, the model group, and the pretreatment model group were 1.00 ± 0.00, 0.85 ± 0.01, and 0.65 ± 0.01, respectively, and the differences between the groups were statistically significant (F = 23.360, 55.350, P < 0.01). The percentages of (F4/80(+)CD86(+)) and (F4/80(+)CD163(+)) in the normal control group and model group and pretreatment model group were 10.67% ± 0.91% and 12.60% ± 1.67%, 20.02% ± 1.29% and 8.04% ± 0.76%, and 43.67% ± 2.71% and 5.43% ± 0.47%, respectively, and the differences among the groups were statistically significant (F = 11.130/8.379, 39.250/13.190, P < 0.01). The liver injury scores of the normal control group, the model group, and the pretreatment model group were 0.22 ± 0.08, 1.32 ± 0.36, and 2.17 ± 0.26, respectively, and the differences among the groups were statistically significant (F = 12.520 and 22.190, P < 0.01). Conclusion: Ces1f may be a hepatic inflammatory inhibitory molecule, and its inhibitory effect production may come from the molecule's maintenance of KC polarization phenotypic homeostasis.
Subject(s)
Animals , Male , Mice , Carboxylesterase/genetics , Galactosamine , Gene Knockdown Techniques , Kupffer Cells , Lipopolysaccharides/adverse effects , Liver Failure, Acute/chemically induced , Mice, Inbred C57BL , RNA, MessengerABSTRACT
Objective To explore the expression feature of long non-coding RNA (lncRNA) 1010001N08Rik in hyperoxia-induced bronchopulmonary dysplasia (BPD) and predict the mechanism that 1010001N08Rik might be involved in the occurrence and development of BPD by a series of bioinformatics analysis.Method The sequence,genomic position and structure characteristics of 1010001N08Rik were acquired from UCSC genome browser,and its target gene was predicted by Ensemble database.We successfully established the animal model of BPD by making newborn C57BL/6J mice exposed to 95％ concentrations of ambient oxygen for seven days.The expression of 1010001N08Rik and Gata 6 were detected using real-time quantitative polymerase chain reaction (PCR).Student's t test was used to compare their expression levels during the BPD process.Result The relative expression of 1010001N08Rik in BPD process at d1,d3,d5,d7 was 1.21 ± 0.33,2.02 ± 0.41,2.95 ± 0.45,4.20-± 0.48 respectively,and there were significant difference between adjacent time points (P ＜ 0.05).The relative expression of Gata 6 mRNA was 0.92 ±0.30,1.10 ± 0.31,0.86 ± 0.24,0.45-± 0.08 respectively,and there was significant difference between d5 and d7 (P ＜0.05).1010001N08Rik had highly conserved property among different species.The chromosomal regions of 1010001N08Rik existed transcriptional factors binding locations and epigenetic regulation clues,and its possible candidate target gene was Gata 6.Conclusion The expression of 1010001N08Rik increased during the formation process of BPD.Bioinformatics analysis and preliminary experiment results suggested that 1010001N08Rik might participate in the process of BPD by down-regulating Gata 6 expression.
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Objective To study the pulmonary surfactant (PS) on prevention of neonatal respiratory distress syndrome (NRDS) in neonates delivered via caesarean section. Methods From selective cesarean section infants (gestational age 34-38+6 W), 80 cases whose test tube oscillation tests were negative and amniotic fluid pulmonary surfactant associated protein A (SP-A) concentrations were lower than <10μg/L, and were randomly divided into PS prevention group and control group, with 40 cases in each group. PS prevention group within 1 h of birth were administrated poractant alfa injection by endotracheal tube (dose 100 mg/kg), but the control group was not given special treatment, leaving only the observation. The incidence of NRDS, treatment status and clinical progression were compared between two groups. Results The incidence of NRDS in control group was 82.5%(33/40), in PS prevention group was 37.5%(15/40), and there was significant difference (P<0.05). The degree of NRDS in control group was more severe. The incidence rate of persistent pulmonary hypertension of the new-born (PPHN), pulmonary air leak, patent ductus arteriosus and oxygenation index above 25 mmHg (1 mmHg=0.133 kPa) in control group were significantly higher than those in PS prevention group (P<0.05). The time of mechanical ventilation, the time of oxygen inhalation, ratio of arterial partial pressure of oxygen (PaO2) before mechanical ventilation to fraction of inspired oxygen (FiO2), and costs of hospitalization in control group were significantly higher than those in PS prevention group (P<0.05). Conclusions PS prevention can reduce the incidence of NRDS of neonates delivered by elective caesarean section, can alleviate the symptoms of NRDS, shorten length of stay and reduce the cost of hospitalization.
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Objective To investigate the role of receptor for advanced glycation end - products nuclear factor - κB(RAGE - NF - κB)signaling pathway in the lipopolysaccharide - induced acute lung injury(ALI)in neo-natal rats. Methods Thirty - two SD rats were divided into 4 groups by complete randomization method(8 cases in each group).(1)Lipopolysaccharide(LPS)group was given intraperitoneal injection of 9 g/ L saline and 3 mg/ kg LPS 1 h later.(2)Bortezomib group was given intraperitoneal injection of Bortezomib(0. 2 mg/ kg)and 3 mg/ kg LPS 1 h later.(3)Anti - RAGE mAb group was given intraperitoneal injection of anti - RAGE mAb(15 mg/ kg)and 3 mg/ kg LPS 1 h later.(4)Control group was given 9 g/ L saline was given at each time point. All the rats were sacrificed and observed 24 h later. Levels of tumor necrosis factor(TNF) - α in the plasma and bronchoalveolar lavage fluid(BALF) were detected by enzyme linked immunosorbent assay. RAGE and NF - κB levels in tissue homogenates were detected by Western blot and mRNA levels were detected by reverse transcription - polymerase chain reaction. The pathological assessment of the lung tissues was performed by HE staining. Results (1)Among 4 groups,there were significantly differences in TNF - α in serum and BALF(F = 150. 70,P ﹤ 0. 001;F = 165. 83,P ﹤ 0. 001). Levels of TNF - α in LPS group were significantly higher than those of two pretreatment groups(all P ﹤ 0. 05).(2)Western blot figures il-lustrated that the concentrations of RAGE mRNA and NF - κB in anti - RAGE mAb group and bortezomib group were lower than those of the LPS group.(3)Reverse transcription - polymerase chain reaction analysis showed that there were significant differences in the expression of RAGE mRNA and NF - κB mRNA among 4 groups(F = 175. 14,P ﹤0. 05;F = 188. 65,P ﹤ 0. 05). Levels of RAGE mRNA and NF - κB mRNA in the LPS group were significantly higher than those of two pretreatment groups(all P ﹤ 0. 05).(4)Lung injury score differences among 4 groups were statistical-ly significant(F = 106. 01,P ﹤ 0. 001). Pathological changes in two pretreatment groups reduced compared to those of the LPS group(all P ﹤ 0. 05). Conclusions RAGE - NF - κB signaling pathway regulates the LPS - induced ALI in neonatal rats. Anti - RAGE mAb and Bortezomib both have a protective effect on LPS - induced ALI.
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Objective To evaluate the effect of Saccharomyces boulardii (SB) administration on very-low-birth-weight (VLBW) infants.Methods One hundred and ninety-eight preterm infants were prospectively randomized into observation group (105 cases) and control group (93 cases) based on the symptomatic and supportive treatment.When uncompletely stomach intestine nutrition fed,the patients of observation group took SB (50 mg/kg),the patients of control group took equivalent placebo.The times of defecation and diarrhea,the rate of neonatal necrotizing enterocolitis,hospital onset of infection (septicemia,pulmonary infection),fungal infection,the time of intravenous nutrition and length of stay were compared.Results The general data in two groups had no significant difference (P ＞ 0.05).The times of defecation,time of intravenous nutrition and length of stay in two groups had significant difference [(1.8 ± 0.4) times/d vs.(3.4 ± 0.5) times/d,(30.21 ± 3.43) d vs.(40.47 ± 4.35) d,(33.5 ± 6.8) d vs.(45.4 ± 9.3) d] (P ＜ 0.05).The rate of diarrhea,neonatal necrotizing enterocolitis,septicemia and pyemia in two groups had significant difference [14.3％ (15/105) vs.25.8％ (24/93),11.4％ (12/105) vs.19.4％ (18/93),19.0％ (20/105) vs.29.0％ (27/93)] (P ＜ 0.05).The rate of pulmonary infection and fungal infection between two groups had no significant difference(P＞ 0.05).Conclusion SB administration on VLBW infants can reduce the infection,promote enteral feeding,shorter hospital stay,and has a certain significance on the family and the community.
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Breast cancer related lymphedema is the accumulation of interstitial fluid due to the lymphatic fluid load exceeds its transport ability as a consequence of the lesion of lymphatic structure.It mainly presents as swelling of the affected limb,a feeling of heaviness or tightness in the limb,aching and restricted limb motion.Its incidence is related to surgery,chemical therapy,obesity,hypertention as well as the alteration of particular genes.To prevent the occurrence and exacerbation of lymphedema,patients should avoid the overuse of the affected limb.While limb exercise step by step can solve part of this problem,but not all,doctor should also take both the patterns of exercise and risk factors that patients expose into consideration,then make a individualized exercise plan,evaluate the condition precisely,in order to achieve the best treatment purposes.
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<p><b>OBJECTIVE</b>To investigate the influence of high oxygen exposure on signaling pathway of the receptor for advanced glycation end products (RAGE)-NF-κB of lung in newborn rats and the mechanisms of protecting lung injury for human mesenchymal stem cells (hMSC).</p><p><b>METHODS</b>Twenty-four newborn Sprague-Dawley rats from three litters were randomly divided into three groups, as hyperoxia exposed + hMSC group (group A), hyperoxia exposed group (group B), and air-exposed group (group C). The rats from the group A and B were placed in a sealed Plexiglas chamber with a minimal in-and outflow, providing six to seven exchanges per hour of the chamber volume and maintaining O(2) levels above 95%, while rats in the group C only exposed to air simultaneously. Seven days later, rats in the group A were injected intravenously with hMSC (5×10(4)) after hyperoxia exposure, but rats in group B and C received subcutaneous injection with PBS alone at the same time point. Then all the rats were exposed to air, and were sacrificed three days later. Immunohistochemistry was used to evaluate the expression of RAGE in lung tissue. The levels of TNF-α and sRAGE in bronchoalveolar lavage fluid (BALF) and in serum were detected by ELASA, RAGE mRNA and NF-κB mRNA in tissue homogenates were detected by RT-PCR, RAGE and NF-κB by Western blotting; also the value of lung damage score were calculated with histology under light microscope.</p><p><b>RESULTS</b>There were significant differences among three groups in the fields of lung damage score (F = 51.59, P = 0.000), mRNA and protein of RAGE (F = 37.21, P = 0.000; F = 15.88, P = 0.000) and NF-κB (F = 5.695, P = 0.011; F = 4.223, P = 0.0288) in lung tissue homogenates, and the level of TNF-α (F = 38.29, P = 0.000) in BALF, all these parameters in group A and group B were higher than that in group C. While sRAGE in BALF in group A and group B were less than that in group C (F = 4.804, P = 0.0191). There were also significant differences between group A and group B in these parameters (P < 0.05). There were also no significant differences neither in TNF-α nor in sRAGE in serum among three groups.</p><p><b>CONCLUSIONS</b>hMSC protects hyperoxia-induced lung injury via downregulating the signaling pathway of RAGE-NF-κB.</p>
Subject(s)
Animals , Female , Humans , Male , Rats , Animals, Newborn , Disease Models, Animal , Glycation End Products, Advanced , Genetics , Metabolism , Hyperoxia , Metabolism , Lung , Metabolism , Pathology , Lung Injury , Metabolism , Mesenchymal Stem Cell Transplantation , NF-kappa B , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Random Allocation , Rats, Sprague-Dawley , Signal Transduction , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the expression of erythropoietin (EPO) and its receptor (EPOR) in the brain of newborn rats suffering fetal distress.</p><p><b>METHODS</b>A model of fetal distress was prepared by ligating bilateral uterine arteries of the rats with full-term pregnancy for 10 minutes before cesarean sections. The expression levels of EPO and EPOR in the brain of newborn rats were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot at 0, 2, 6, 12, 24, 48, 72 hrs and 7 days after birth. Serum EPO levels were measured using ELISA simultaneously. The newborn rats born by cesarean sections which were not subjected to uterine artery ligation were used as the control group.</p><p><b>RESULTS</b>The expression of EPO protein and mRNA in brain tissues in the fetal distress group increased significantly compared with the control group 2, 6 and 12 hrs after birth (P<0.05). The expression of EPOR protein and mRNA in brain tissues in the fetal distress group increased significantly compared with the control group 2, 6, 12, 24 and 48 hrs, and 3 days after birth (P<0.05). Serum EPO levels in the fetal distress group were significantly higher than in the control group 2 hrs after birth.</p><p><b>CONCLUSIONS</b>The EPO and EPOR levels in the brain increase quickly after birth in newborn rats suffering from fetal distress. The EPOR is high expressed for a longer time than EPO. This can provide a basis for the treatment of neonatal brain damage induced by fetal distress by exogenous EPO.</p>
Subject(s)
Animals , Female , Pregnancy , Rats , Animals, Newborn , Brain , Metabolism , Pathology , Erythropoietin , Blood , Genetics , Fetal Distress , Metabolism , RNA, Messenger , Rats, Sprague-Dawley , Receptors, Erythropoietin , Blood , GeneticsABSTRACT
Objective To observe influence of retinoic acid on the lung damage and expression of Connexin26(Cx26) of neonatal rats suffered from hyperoxia.Methods Twenty-four 3-day SD rats were randomly divided into three groups:retinoic acid+ hyperoxia,hyperoxia,air group;rats in the group of hyperoxia were suffered 7 days hyperoxia (the level of oxygen was above 950 mL/L),air group was instead of air.All rats were weighed,value of radical alveolar counts(RAC) and microstructure of the lungs were observed by HE staining under light and electron microscope respectively;and the expression of Cx26 in the lungs was determined by immunohistochemistry.Results Compared with air group,weight,value of RAC were both decreased obviously in hyperoxia,retinoic acid+hyperoxia groups[(13.53?1.27),(13.38?1.29),(17.37?0.89)g,F=30.19 P=0;11.15?1.33,12.49?1.47,13.94?0.98,F=9.59 P=0),while the expression of Cx26 increased [(13.68?1.28)%,(17.82?1.72)%,(19.69?1.77)%,F=29.36 P=0].Conclusion Retinoic acid has a protective effection from damaged lung of neonatal rats suffered from hyperoxia,and it may be related to the alteration of expression of Cx26 protein in the lungs.