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Objective:To explore the correlation between carotid lesion severity detected by ultrasound and pregnancy associated plasma protein A (PAPP‐A) expression in patients with acute coronary syndrome (ACS) .Methods :A to‐tal of 78 ACS patients hospitalized in our hospital from Feb 2012 to May 2015 were regarded as ACS group ,mean‐while ,another 78 healthy subjects were enrolled as healthy control group .Correlation between carotid lesion severi‐ty detected by ultrasound and PAPP‐A expression was analyzed . Results:Compared with healthy control group , there were significant rise in serum PAPP‐A concentration [ (0.97 ± 0.32) mg/L vs .(1.56 ± 0.19) mg/L] ,carotid intima‐media thickness [IMT ,(0.84 ± 0.13) mm vs .(1.28 ± 0.16) mm] and Crouse plaque score [ (2.98 ± 1.92) scores vs .(8.24 ± 1.13) scores] in ACS group ,P<0.01 all .Linear correlation analysis indicated that serum PAPP‐A concentration was significant positively correlated with Crouse plaque score and IMT ( r= 0.342、0.243 , P<0.05 all) .Multi‐factor gradual linear regression analysis indicated that carotid Crouse plaque score and IMT were in‐dependent risk factors for PAPP‐A (partial regression coefficient=1.932 ,17.722 ,P<0.01 both) .Conclusion:Ca‐rotid ultrasound Crouse plaque score ,IMT are significantly positively correlated with PAPP‐A expression ,which can indirectly reflect coronary artery disease severity in ACS patients ,it is worth extending .
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Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.
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Objective To investigate the expression of placental growth factor (PlGF) protein and mRNA in lungs of neonatal rats exposed to 85%hyperoxia, and to establish the relationship between PlGF and bronchopulmonary dysplasia (BPD). Methods Forty-eight Sprague–Dawley neonatal rats were randomly exposed to air (control group)(n=24) and 85% hyperoxia (hyperoxia group)(n=24)within 12 h after birth. The rats were sacrificed at 3, 5 and 7 days after exposure (eight at each time) and their lungs were sampled. PlGF protein and mRNA expression in the lungs were determined by Western blot and real-time polymerase chain reaction (PCR) at 3, 5 and 7 days. Left lung tissue was used for morphological and histological observation with hematoxylin and eosin staining. Terminal air spaces and the secondary septa were counted manually under microscope. T-test was applied for statistics. Results Compared with the control group, morphological and histological analysis in the hyperoxia group revealed inflammatory cell infiltration, simplified alveolar structure, less alveolar, alveolar cavity expansion and thickened alveolar septum. Morphometric measurements showed that terminal air spaces and secondary septa were significantly fewer in the hyperoxia rats than those in the control group at 5 and 7 days (terminal air spaces:23.6±8.2 vs 33.1±6.2 and 28.5±9.2 vs 38.4±10.1, t=1.91, 2.53, all P<0.05;secondary septa:56.0±12.2 vs 78.3±8.2 and 75.4±12.2 vs 126.1±10.2, t=2.14, 2.72, all P<0.05). Real-time PCR showed that expression of PlGF mRNA increased significantly on day 3, 5 and 7 in the hyperoxia group compared with the control group (1.16±0.17, 1.34±0.15 and 1.65±0.19 vs 0.65±0.21, 0.47±0.11 and 0.46±0.17, respectively, t=1.93, 2.55, 2.79, all P<0.05). Western blot also showed that expression of PlGF protein on day 3, 5 and 7 in the hyperoxia group increased compared with the control group, but only being significant on day 3 (0.24±0.17 vs 0.09±0.01, t=2.44, P<0.05). Conclusions Hyperoxia (85%) exposure could increase PlGF protein and mRNA expression in the lungs of neonatal rats, likely contributing to pathogenesis of BPD, and might lead to pulmonary vascular developmental disorders in BPD.
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Objective To investigate the effect of extracellular signal-regulated kinase(ERK)pathway OH nitriC oxide(NO)release by human umbilical vein endothelial cell(HUVEC)induced by placental growth factor-1(PLGF-1).Methods During January to April 2006,50 samples of umbilical vein blood were collected from newborns delivered by cesarean section due to intrauterine distress and abnormal fetal position.HUVEC were primarily cultured by trypsin digestion.Then the following procedures were performed:(1)Cells were identified using the morphology andⅧfactor immunohistochemistry methods if the culture WaS satisfactory.(2)Cells were collected,and fms.1ike tyrosin kinase(Fit-1)protein and its mRNA expression were detected with immunoprints and RT-PCR methods.(3)The protein wag extractedafter cells were treated with PLGF-1(cells were collected before the treatment and 2.5,5.10,20 min after the treatment).The protein levels of ERK were determined by immunoprints.(4)The cells were cultured witll serum-free culture medim containing PLGF-1 only(culture media were collected 20,40,160,360.480.720 and 1440 rain after the treatment).The quantity of NO was detected with nitrate reductase metllod(5)The ceHs were cultured with serum.free culture medium containing PI)98059,the inhibitor of mitogen-activated protein kinase(MAPK)/MEK for 60 min Then the cells were cultured continuously with the serum-free culture medium containing PLGF-1 for 60 mira The culture media were coilected.The quantity of NO was detected by nitrate reductase method.The samples were divided into treatment group and control group.Control group was exactly the satne in treatment time,culture condition,and time to colleet the cells as the treatment group.except that it WaS not treated with PLGF-l or PD 98059.Resuits (1)By morphology and Ⅷ factor inununohistochemistry the cultured cells were identified to be HUVEC.(2)Fit-1mRNA and protein were expressed in HUVEC.(3)Expression of ERK protein started to increaSe at 2.5 min after treatment of HUVEC with PLGF-1,reaching the peak at 5 min,and decreased at 10 min.(4)Incomparison with the control group.NO started to increase at 20 rain after treatment of HUVEC with PLGF-lat 480 min(15.82±0.69)μmol/L Comparison between the two groups showed a significant difference (P<0.05).(5)ReleaSe of NO from the cells treated with PD98059 for 1 hour and PLGF WaS significantly inhibited,compared with the ceils treated with PLGF-1 only.Conclusion ERK pathways play an important role in N0 release bv HUVEC induced by PLGF.
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Objective To investigate the expression of Fas antigen (Fas) and ligand (FasL), placental growth factor (PlGF) in placenta of pregnant women with pre-eclampsia. Methods Expressions of Fas, FasL and PlGF of placenta were determined by immunohistological streptavidin-peroxidase-biotin (SP) method and compared between normal late pregnancy (24 cases) and mild pre-eclampsia (24 cases) and severe pre-eclampsia (24 cases) groups. Results Different expression of levels Fas, FasL and PlGF were observed in the trophoblasts of most placentae. Positive immunostain of Fas, FasL and PlGF was mainly located in the cytoplasm and membrane of syncytiotrophoblast. FasL and PlGF expressions (63?4, 81?6 and 42?6, 65?6) were significantly less (P
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Objective To investigate the expression pattern of Placental growth factor (PlGF) and regulatory effects of interleukin-6 (IL-6)on PlGF expression in the primary cultured trophoblast cells. Methods Cytotrophoblast cells werecollected by trypsin-collagenase digestion and Percoll gradient centrifugation for primary culture. Reverse transcriptase polymerase chain reaction(RT-PCR) were resorted to demonstrate the expression pattern of PlGF mRNA in trophoblast cells cultured in vitro. The effects of IL-6 with different concentration(100,10,1 and 0.1 ?g/L) and duration (6,12,24 and 48 h) on PlGF expression were observed. Results The 185 bp and 248 bp bands of PlGF were showed by RT-PCR. PlGF expression correlated with IL-6. PlGF began to increase at 6 h, and reached the climax at 12 h when recultured with 100 ?g/L IL-6. Conclusions PlGF expression has time and dose dependance on IL-6. It may play an important role in early pregnancy.