Relationship between Sestrin2 and mtDNA-NLRP3 inflammasome pathway during endotoxin-induced myocardial injury in mice / 中华麻醉学杂志

Objective:To evaluate the relationship between Sestrin2 and mitochondrial DNA (mtDNA)-NOD-like receptor associated protein 3 (NLRP3) inflammasome pathway during endotoxin-induced myocardial injury in mice.Methods:One hundred and eighty-four clean-grade healthy male ICR mice, aged 8-12 weeks, weighing 20-25 g, were used in this study. One hundred and sixty-eight mice were divided into 7 groups ( n=24 each) using the random number table method: normal control group (N group), lipopolysaccaride(LPS) group (L group), mtDNA group, LPS+ mtDNA group (M group), normal control+ negative control adeno-associated virus (AAV-NC)group (NC group), LPS+ mtDNA+ AAV-NC group (MC group), and LPS+ mtDNA+ Sestrin2 overexpression adeno-associated virus (AAV-Sestrin2) group (MSgroup). Another 10 mice were used to detect the transfection effect of AAV-Sestrin2, and the left 6 mice were used for mtDNA extraction. The model of endotoxemia was developed by intraperitoneal injection of LPS 10 mg/kg. mtDNA 5 mg/kg was intraperitoneally injected in mtDNA group, and mtDNA 5 mg/kg was intraperitoneally injected at 30 min after LPS injection in M group.AAV-Sestrin2 150 μl was injected via the tail vein in MS group, and the equal volume of AAV-NC was injected via the tail vein in MC and NC groups. Four weeks after virus injection, LPS 10 mg/kg was intraperitoneally injected and 30 min later mtDNA 5 mg/kg was intraperitoneally injected in MS and MC groups. Blood samples were collected at 24 h after LPS injection for determination of serum creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) activities (by biochemical assay), concentrations of serum cardiac troponin I (cTnI), interleukin-18 (IL-18) and interleukin-1beta (IL-1β)(by enzyme-linked immunesorbent assay), and expression of mtDNA (by quantitative real-time polymerase chain reaction). The animals were sacrificed after the end of blood sampling and myocardial tissues were obtained for determination of the contents of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), and adenosine triphosphate (ATP) and expression of NOD-like receptor associated protein 3 (NLRP3), active subunit p20 of caspase-1 (caspase-1p20) and apoptosis-associated microprotein (ASC) in myocardial tissues (by Western blot) and for microscopic examination of the pathological changes after HE staining (with a light microscope). Results:Compared with N group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in L group and mtDNA group.Compared with L group and mtDNA group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly increased, the expression of mtDNA was up-regulated, the ROS content in myocardial tissues was increased, the T-AOC and ATP contents in myocardial tissues were decreased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was up-regulated( P<0.05), and the pathological changes of myocardial tissues were aggravated in M group. Compared with M group, the levels of CK-MB, LDH, cTnI, IL-1β and IL-18 in serum were significantly decreased, the expression of mtDNA was down-regulated, the ROS content in myocardial tissues was decreased, the T-AOC and ATP contents in myocardial tissues were increased, the expression of NLRP3, caspase-1p20 and ASC in the myocardial tissues was down-regulated( P<0.05), and the pathological changes of myocardial tissues were significantly attenuated in MS group. Conclusions:Sestrin2 can reduce endotoxin-induced myocardial injury in mice by alleviating mitochondrial damage, inhibiting oxidative stress, protecting mtDNA from oxidative damage, and then inhibiting mtDNA-NLRP3 inflammasome pathway.

In situ isolation of nuclei or nuclear proteins from adherent cells: a simple, effective method with less cytoplasmic contamination

BACKGROUND: Isolation of nuclei or nuclear proteins is a prerequisite for western blot, nuclear proteome profiling, and other evaluations of nuclear proteins. Here, we developed a simple method for in situ isolation of nuclei or nuclear proteins by in situ removing the extranuclear part of adherent cells via a classical nonionic detergent triton X-100. RESULTS: First, the feasibility of our method was confirmed by confocal microscopy, atomic force microscopy, scanning electron microscopy, dynamic light scattering, immunofluorescence imaging, and time-lapse dynamic observation. Next, the optimal concentration range (approximately 0.1-1% for ~ 10 min) of triton X-100 and the optimal treatment time (< 30 min) of 0.1-1% Triton X-100 for our method were determined via western blotting of eight extra-/ intra-nuclear proteins. Subsequently, the effectiveness, sensitivity, and cytoplasmic contamination of our method were tested by investigating the levels of phosphorylated p65 (a NF-κB subunit) in the nuclei of endothelial or tumor cells treated with/without lipopolysaccharide (LPS) via western blotting and by comparing with a commercial nuclear protein extraction kit (a classical detergent-based method). The data show that compared with the commercial kit our method obtained a higher yield of total nuclear proteins, a higher pP65 level in both control and LPS groups, and much lower content of GAPDH (as a reference for cytoplasmic contamination) in nuclei. CONCLUSIONS: The in situ isolation of nuclei or nuclear proteins from adherent cells in this study is a simple, effective method with less cytoplasmic contamination. This method/strategy has the potential of improving the quality of downstream evaluations including western blotting and proteomic profiling.

Pedigree analysis and prenatal diagnosis of intellectual disability caused by synonymous mutations of p.S434S in DLG3 gene / 中华围产医学杂志

Objective:To analyze the pathogenic gene and prenatal diagnosis of a family with intellectual disability.Methods:Out of this family consisting of 17 members in three generations, four males had intellectual disability. The proband's elder sister (Ⅱ-7) visited Henan Provincial People's Hospital in Oct 2019 for genetic counseling at 8 weeks of gestation. After informed consent was obtained, peripheral blood samples of the family members were collected. The whole exome sequencing was performed on the genome DNA of the proband (Ⅱ-9, male) and his parents to screen the candidate variants for phenotype co-segregated analysis by Sanger sequencing. The expression vectors were constructed by homologous recombination and the splicing experiments were performed in vitro. Reverse transcription polymerase chain reaction, Sanger sequencing, and TA clone sequencing were used to analyze the effect of candidate variants on splicing. After the pathogenic variant was determined the proband's elder sister underwent prenatal diagnosis (Ⅲ-7) using goldeneyeTM20A genotyping system and Sanger sequencing. Results:A hemizygous synonymous variant of c.1302G>A (p. S434S) in DLG3 gene was found in the proband by whole exome sequencing, which was carried by his mother (Ⅰ-1) and co-segregated with the phenotype in other family patients. In vitro splicing experiment showed that c.1302G>A variant led to abnormal splicing of 88.24% transcripts, which further resulted in the reading frame shift and protein function impairment. The mutation was not detected in the fetus (Ⅲ-7), who was born alive later and showed no abnormal mental or behavioral development at the age of one and a half year and is still being followed up. Conclusions:The synonymous mutation c.1302G>A in DLG3 gene was the etiopathogenesis of X-linked intellectual disability in this family.

Role of SIRT1 in electroacupuncture-induced reduction of central post-stroke pain in rats: relationship with NLRP3 / 中华麻醉学杂志

Objective:To evaluate the role of silent information regulator 1 (SIRT1) in electroacupuncture (EA)-induced reduction of central post-stroke pain (CPSP) and the relationship with nod-like receptor pyrin domain containing 3 (NLRP3) in rats.Methods:Fifty SPF healthy male Sprague-Dawley rats, aged 6 weeks, weighing 180-220 g, were divided into 5 groups ( n=10 each) using a random number table method: sham operation group (group Sham), CPSP group, CPSP+ sham EA group (group SEA), CPSP+ EA group (group EA) and CPSP+ EA+ SIRT1 inhibitor EX527 group (group EX527). Type Ⅳ collagenase was injected into the right ventral posterolateral nucleus to establish the model of CPSP in CPSP, SEA, EA and EX527 groups.At 24 h after the model was established successfully, 30 min EA (frequency 2/15 Hz) stimulation of Neiguan, Renzhong and Sanyinjiao was performed once a day for 5 consecutive days in EA group.EA was performed at the points 5 mm lateral to the acupoints of Neiguan, Renzhong and Sanyinjiao in group SEA, and the other procedures were similar to those previously described in group EA.SIRT1 inhibitor EX527 5 mg/kg was injected intraperitoneally at 30 min before EA stimulation in group EX527, and the other procedures were similar to those previously described in group EA.At 1 day before the establishment of model (T 0) and at 1, 3 and 5 days after the establishment of model (T 1-3), the thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured.The animals were then sacrificed and brain tissues were taken for determination of the expression of SIRT1, NLRP3 and interleukin (IL)-18 and IL-1β. Results:Compared with Sham group, the TWL was significantly shortened and the MWT was decreased at T 1-3, the expression of SIRT1 was down-regulated, and the expression of NLRP3, IL-18 and IL-1β was up-regulated in CPSP, SEA, EA and EX527 groups ( P<0.05). Compared with CPSP group, the TWL was significantly prolonged and the MWT was increased at T 1-3, the expression of SIRT1 was up-regulated, and the expression of NLRP3, IL-18 and IL-1β was down-regulated in EA group ( P<0.05), and no significant change was found in the parameters mentioned above in group SEA ( P>0.05). Compared with EA group, the TWL was significantly shortened and the MWT was decreased at T 1-3, the expression of SIRT1 was down-regulated, and the expression of NLRP3, IL-18 and IL-1β was up-regulated in EX527 group ( P<0.05). Conclusion:SIRT1 is involved in the process of EA-induced reduction of CPSP, which is related to inhibiting NLRP3 expression in rats.

Effect of nucleolar protein 14 on angiogenesis in melanoma and its mechanism / 中华皮肤科杂志

Objective To evaluate the effect of nucleolar protein 14 (NOP14) on angiogenesis in melanoma.Methods Melanoma tissues were collected from 40 patients with pathologically diagnosed melanoma in Guangzhou First People's Hospital from January 2016 to December 2018,and immunohistochemical study was conducted to determine the expression of NOP14 and CD31 (expressed as microvessel density [MVD]).Melanoma cell lines A375 and SK-MEL-1 were both divided into 4 groups:empty vector group transfected with the empty vector,NOPI4 group transfected with a NOP14-overexpressing vector,siNOP14 group transfected with the siRNA targeting NOP14,and siNC group transfected with a negative control siRNA.Fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of NOP14 respectively,and Western blot analysis and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in cells and their culture media.Coculture models of human umbilical vein endothelial cells (HUVECs) and A375/SK-MEL-1 cells in the above groups were established in Transwell chambers,and cell counting kit-8 (CCK8) assay,Transwell migration and invasion assays and Matrigel-based vasculogenic mimicry assay were performed to evaluate the cellular proliferative,migratory,invasive activity and tube formation capacity respectively.A linear regression model was used to analyze the relationship between NOP14 expression and MVD in melanoma tissues,multi-way analysis of variance to analyze the difference in cellular proliferative activity,and independent-sample t test to compare other experimental indices between 2 groups.Results The expression of CD31 (MVD) was 44 ± 13 in the group with high NOP14 expression (n =20),58 ± 16 in that with moderate NOP14 expression (n =17),and 62 ± 11 in that with low NOP14 expression (n =3).The NOP14 expression was negatively correlated with MVD (r =-0.525,P =0.017).Compared with the empty vector group,the expression of VEGF and VEGFR in A375 and SK-MEL-1 cells and their culture media significantly decreased in the NOP14 group (all P ＜ 0.05).Compared with the siNC group,the expression of VEGF and VEGFR in the A375 and SK-MEL-1 cells and their culture media significantly increased in the siNOP14 group(all P ＜ 0.05).In the co-culture models of A375 cells and HUVECs,the NOP14 group showed significantly decreased proliferative activity of HUVECs (F =131.85,P ＜ 0.05),and numbers of migratory cells (22 ± 5 vs.63 ± 8,t =7.07,P =0.002),invasive cells (14 ± 5 vs.45 ± 10,t =4.94,P =0.008) and branch points (8 ± 2 vs.14 ± 3,t =5.06,P ＜ 0.001) compared with the empty vector group;compared with the siNC group,the siNOP14 group showed significantly increased proliferative activity of HUVECs (F =79.92,P ＜ 0.01),and numbers of migratory cells (152 ± 30 vs.59 ± 4,t =5.36,P =0.006),invasive cells (134 ± 21 vs.50 ± 8,t =6.40,P ＜ 0.001) and branch points (27 ± 3 vs.15 ± 4,t =6.10,P ＜ 0.001).In the co-culture models of SK-MEL-1 cells and HUVECs,the 4 groups showed the same trend of changes in the cellular proliferative,migratory,invasive activity and tube formation capacity of HUVECs as the above groups in the co-culture models of A375 cells and HUVECs.Conclusion The NOP14 expression is negatively correlated with MVD in melanoma tissues,and NOP14 can inhibit angiogenesis in melanoma.

Establishment of electrical immunosensor for the detection of nuclear matrix protein-22 / 中华检验医学杂志

Objective@#A label-free electrochemical immunosensor was developed for the detection of nuclear matrix protein-22 (NMP22) as a biomarker of bladder cancer.@*Methods@#The study was based on the establishment and validation of the methodology. Urine samples were collected from 20 patients with bladder cancer and 20 controls in the affiliated Hongqi hospital of Mudanjiang medical university from September in 2017 to July in 2019 to validate the developed method. A screen-printed electrode (SPE) was modified with a film of a composite made from the reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) immobilized Zn-based-Metal-organic frameworks deposited with Au nanoparticles (rGO-TEPA@Au-ZIF8). Primary antibody against NMP22 was immobilized on the Au nanoparticles on the surface of the modified SPE, which then was blocked with bovine serum albumin to elimiate nonspecific binding sites. The process of the construction of the proposed sensorwas characterized by cyclic voltammetry and electrochemical impedance spectroscopy. Differential pulse voltammetry was used to evaluate the linear range, recovery, precision, selectivity and stability. The data were analyzed by Mann-Whitney U test.@*Results@#Under optimal conditions, the immunosensor exhibited a linear range of 0.01-1000 ng/mlwith a detection limit of 3.33 pg/ml (S/N=3) and a standard recovery of 97.65%-107.05%. The levels of NMP22 in urine samples from patients with bladder cancer [66.03 (4.34, 91.74)]ng/ml determined by the proposed sensor were significantly higher than those of controls 0.54(0.06, 8.84) ng/ml(P=0.001).@*Conclusion@#The immunosensor can achieve sensitive, rapid and acucurate detection of NMP22, and has potential application prospects in monitoring tumor markers.

Establishment of electrical immunosensor for the detection of nuclear matrix protein-22 / 中华检验医学杂志

Objective A label-free electrochemical immunosensor was developed for the detection of nuclear matrix protein-22 (NMP22) as a biomarker of bladder cancer. Methods The study was based on the establishment and validation of the methodology. Urine samples were collected from 20 patients with bladder cancer and 20 controls in the affiliated Hongqi hospital of Mudanjiang medical university from September in 2017 to July in 2019 to validate the developed method. A screen-printed electrode (SPE) was modified with a film of a composite made from the reduced graphene oxide-tetraethylene pentamine (rGO-TEPA) immobilized Zn-based-Metal-organic frameworks deposited with Au nanoparticles (rGO-TEPA@Au-ZIF8). Primary antibody against NMP22 was immobilized on the Au nanoparticles on the surface of the modified SPE, which then was blocked with bovine serum albumin to elimiate nonspecific binding sites. The process of the construction of the proposed sensorwas characterized by cyclic voltammetry and electrochemical impedance spectroscopy. Differential pulse voltammetry was used to evaluate the linear range, recovery, precision, selectivity and stability. The data were analyzed by Mann-Whitney U test. Results Under optimal conditions, the immunosensor exhibited a linear range of 0.01-1000 ng/mlwith a detection limit of 3.33 pg/ml (S/N=3) and a standard recovery of 97.65％-107.05％. The levels of NMP22 in urine samples from patients with bladder cancer [66.03 (4.34, 91.74)]ng/ml determined by the proposed sensor were significantly higher than those of controls 0.54(0.06, 8.84) ng/ml(P=0.001). Conclusion The immunosensor can achieve sensitive, rapid and acucurate detection of NMP22, and has potential application prospects in monitoring tumor markers.

Application of serum nucleobindin-2 in the diagnosis of nasopharyngeal carcinoma / 中国医师杂志

Objective To investigate the expression level of nucleobindin-2 (NUCB2/Nesfatin-1) in nasopharyngeal carcinoma (NPC) patients, other patients with head and neck cancer, rhinitis patients, and healthy subjects in the serum, and evaluate the clinical application value of NUCB2/Nesfatin-1 combined with three items of Epstein-Barr (EB) virus (EA-IgA;VCA-IgA;Rta-IgG) in NPC diagnosis.Methods From Xiangya Hospital of Central South University during January 2017 to June 2017, nasopharyngeal carcinoma patients, other patients with head and neck cancer, rhinitis patients, and healthy subjects samples were 140 cases, respectively.The corresponding kits were used to detect nesfatin-1, EA-IgA, VCA-IgA, and Rta-IgG.Results The serum level of nesfatin-1 in NPC patients was the highest, and the difference was statistically significant compared to other groups.The serum levels of EA-IgA, VCA-IgA, and Rta-IgG were the highest in NPC patients, and were significantly different from those in other groups.Nesfatin-1 was significantly correlated with VCA-IgA and Rta-IgG.There was no significant correlation between nesfatin-1, EA-IgA and NPC staging;Multiple linear regression analysis was used to analyze the influencing factors of nesfatin-1, and the regression equation was Y =208.029 + 17.96X1 + 146.702X2 + 398.879X3 (X1:age;X2:VCA-IgA;X3:Rta-IgG;R2 =0.236).Receiver operating characteristic (ROC) curve analysis showed that nesfatin-I combined with VCA-IgA and Rta-IgG had the best efficiency in the diagnosis of NPC.In single index evaluation, nesfatin-1 has the lowest specificity, but the highest sensitivity.Conclusions The sensitivity of nesfatin-1 for NPC diagnosis is polar altitude, it can make up for the deficiency of EA-IgA, VCA-IgA, and Rta-IgG in diagnosing NPC.The combination of VCA-IgA and Rta-IgG can greatly improve the ability of diagnosing NPC.

Expression of nucleolar protein 14 and CD31 in pancreatic cancer mouse model and its correlation with tumor progression / 中华外科杂志

Objective@#To investigate expression of nucleolar protein 14(NOP14) and CD31 in pancreatic cancer mouse model and its correlation with tumor progression.@*Methods@#Clinicopathological data of 5 patients with pathologically confirmed pancreatic ductal adenocarcinoma(PDAC) and hepatic metastasis between January 2013 and December 2015 was collected in Department of General Surgery, Peking Union Medical College Hospital. Immunohistochemistry staining was employed to detect the expression of NOP14 in matched primary PDAC and relevant metastasis.Pancreatic cancer cells with NOP14 stably knocked down were established by transfecting lentivirus with NOP14 targeted silencing RNA.The inhibition efficacy was detected by quantitative real time PCR and western blot.Microvascular density(MVD) in pancreatic cancer transplantation mouse model was determined by CD31 immunohistochemistry staining analysis and correlated with NOP14 expression and tumor progression.@*Results@#NOP14 had a significant higher expression in liver metastasis than primary pancreatic adenocarcinoma (2.09±0.45 vs. 1.31±0.27, P=0.028). NOP14 was knocked down 86 percent on mRNA level determined by qPCR and 78 percents on protein level detected by western blot. MVD was significantly decreased in NOP14-inhibited tumor from both pancreatic cancer cells subcutaneously and orthotopically grafted tumor mouse model with the value of 61.40±13.85 vs. 85.53±14.59 (P=0.041) and 38.33±10.91 vs. 59.33±15.37(P =0.037), respectively. Besides, MVD was positively associated with tumor volume(r=0.842, P<0.01) and metastasis (r=0.726, P=0.008).@*Conclusion@#NOP14 presents higher expression in hepatic metastasis of pancreatic adenocarcinoma and might promote tumor progression by increasing microvascular density.

Diagnostic value of combined detection of autoantibodies against p53 and Bmi-1 in lung cancer / 中国医师杂志

Objective To evaluate the diagnostic value of combined detection of serum autoantibodies against against p53 and Bmi-1 in lung cancer (LC).Methods Serum levels of autoantibodies against p53 and Bmi-1 were measured by enzyme-linked immunosorbent assay (ELISA) in 92 patients with LC and 80 normal controls.The combined diagnostic value was evaluated with the receiver operating characteristic (ROC) curve.Results The serum levels of autoantibodies against p53 and Bmi-1 were significantly higher in LC than in normal controls (0.438 ±0.705 vs 0.045 ±0.035,Z =-7.667,P ＜0.01;0.117±0.061 vs 0.068 ±0.031,Z =-7.179,P ＜0.01).The levels of autoantibodies against p53 and Bmi-1 were not related to age,gender,pathological classification,lymph node status or tumor-node-metastasis (TNM) stage (P ＞ 0.05).The combined detection of two autoantibodies provided an enhanced sensitivity of 63.0％,a specificity of 91.2％ and an area under curve (AUC) of 0.881,which showed better diagnostic efficiency compared to individual autoantibodies.Conclusions Combined detection of autoantibodies against p53 and Bmi-1 shows good diagnostic value,which may aid diagnosis of LC.

Expression and significance of nuclear factor-κB-related proteins in idiopathic orbital inflammatory pseudotumor / 中华实验眼科杂志

Background Idiopathic orbital inflammatory pseudotumor (IOIP) is a commom orbital disease,with serious eye symptoms and replase tendency,and its pathogenesis is still unclear.Nuclear factor-κB (NF-κB)-related proteins participate in many important pathophysiological process,however,whether NF-κB plays a role in the IOIP process is worthy of attention.Objective This study was to explore the roles of NF-κB pathway in IOIP pathogenesis.Methods Twenty-four IOIP specimens were collected during surgery in Beijing Tongren Hospital from September 2010 to May 2016.The histopathological characteristics of IOIP were examined by hematoxylin and eosin staining.The expression and location of NF-κB/p65,p-p65,p50 and inhibitor of κB (IκB-ot) were detected by immunohistochemistry and verified by immunocytochemistry and Western blot assay.Results The histopathological features of IOIP were numerous small lymphocyte infiltraion and fibrous tissue proliferation,and a lot of epithelioid cells were seen in lacrimal gland-involved specimens.NF-κB/p65 was positively expressed in the cytoplasm of all 24 specimens and the nucleus in 15 specimens with the expressing rate of 62.5％.p50 was expressed in the cytoplasm in 22 specimens with the expressing rate of 91.7％ and in the nucleus in 17 specimens with the expressing rate of 70.8％.The positive expression of p-p65 was found in 22 specimens with the expressing rate of 91.7％,and IκB-α was expressed in the cytoplasm of 11 specimens with the expressing rate of 45.8％.These results were confirmed by immunocytochemistry and Western blot assay.Conclusions NF-κB pathway is activiated during IOIP process,and NF-κB pathway may be involved in the pathogenesis of IOIP.

Impact of biological function on ovarian clear cell carcinoma ES2 cell line with ARID1A gene expression down-regulating in vitro / 中华妇产科杂志

Objective To investigate the efficiency of biological function of AT rich interaction domain 1A (ARID1A) gene silenced by small interfering RNA (siRNA) on ovarian clear cell carcinoma ES2 cell line. Methods (1) The three pairs ARID1A gene siRNA interference fragments siN1 (ARID1A-705), siN2 (ARID1A-1513), siN3 (ARID1A-2282) and one pair negative control were respectively designed, and transfected into ES2 cells by RNA interference max reagent transiently. Reverse transcription (RT)-PCR and western blot methods were used to detect the expression of ARID1A mRNA and protein in ES2 cells transfected with interference fragments respectively. So as to select the best silencing effect of siRNA interference fragment(that was siN3),and then was used in the following experiment. (2) The following experiment were divided into three groups, namely siN3 transfection group, negative control group and blank control group. The proliferative activity of three groups of cells after transient transfection ( 6, 24, 48, 72, 96 hours) was assessed by cell counting kit-8 (CCK-8) assay and expressed as absorbance (A) value; the apoptosis rate of three groups of cells transfected transiently with interference fragment was measured by flow cytometry with annexin V/propidium iodide (PI) staining;the ability of cellular invasion of three groups of cells transfected transiently with interference fragment was tested by transwell experiment;the expression of nuclear factor-kappa B (NF-κB), membrane type-1 matrix metalloproteinase (MT1-MMP) and matrix metalloproteinase-2 (MMP2) protein in ES2 cells transfected transiently with interference fragment was measured by western blot. Results (1) The RT-PCR results showed that the ARID1A mRNA relative expression levels in ES2 cells after transfected transiently with siN1, siN2 and siN3 were 0.007 8±0.005 7, 0.006 8±0.000 3 and 0.002 8±0.000 3 respectively. They were all apparently lower than that in the negative control group (0.034 6 ± 0.001 3;all P0.05). However, the proliferative activity of siN3 transfection group cells after transfected transiently at 24, 48, 72, 96 hours were higher than those in negative control group and blank control group (all P<0.01). The flow cytometry results showed that the apoptosis rate of siN3 transfection group cells was (20.0±3.9)%, which was significantly lower than those in negative control group and blank control group [(31.5 ± 5.0)%, (34.0 ± 4.2)%, respectively;all P<0.05]. The transwell experiment showed that the penetrated cell counts of siN3 transfection group was 60.4±2.9, which was apparently higher than those in negative control group and blank control group (54.2 ± 3.5, 52.1 ± 3.8, respectively; all P<0.01). Western blot experiment showed that the relative expression levels of NF-κB, MT1-MMP and MMP2 protein in siN3 transfection group were respectively 1.85 ± 0.16, 0.37 ± 0.08, 1.38 ± 0.11, which were apparently higher than those in negative control group (0.93±0.11, 0.17±0.05, 0.86±0.06;all P<0.05) and blank control group (0.94 ± 0.04, 0.15 ± 0.08, 0.85 ± 0.10, respectively; all P<0.01). Conclusions It would be to promote the cell doubling time, reduce cell apoptosis and increase the invasive capability in ES2 cells that ARID1A expression was down-regulating by ARID1A mRNA interference. The invasion mechanism may be related to the activation of NF-κB signal transduction pathway, up-regulation of MT1-MMP expression and then promoting the invasion of tumor cells via the up-regulation of MMP2 expression.

Fetal lung structures and expression of pulmonary surfactant proteins of B and C and their modulating factors in rat with gestational diabetes mellitus / 中华围产医学杂志

Objective To explore the lung developmental disorder of rats with gestational diabetes mellitus (GDM) via investigating the GDM rat fetal lung structures and expression of pulmonary surfactant proteins (SP)-B,SP-C,thyroid transcription factor (TTF)-1 and pleiomorphic adenoma gene like (PLAGL)-2.Methods Sprague-Dawley rats were used to construct the GDM model.Twenty GDM rats were used as GDM group and 20 normal pregnant rats as control group.Cesarean section was performed on day 21 of gestation and random blood sugar was detected,and fetal rats were counted and weighed.Ultrastructure of the fetal lungs was studied by transmission electron microscopy.Sixty fetal rats were selected randomly in each group,and 360 paraffin sections were made from fetal lungs.One hundred discontinuous paraffin sections were picked up in each group to observe morphological and structural changes under optical microscope.The other one hundred discontinuous paraffin sections were picked up in each group to detect the location and expression of SP-B,SP-C,TTF-1 and PLAGL-2 protein by immunohistochemistry.Nine fetal rats were selected randomly to detect the expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in fetal lung tissues by Western blotting.Twenty seven fetal rats were selected randomly to detect the mRNAs level of SP-B,SP-C,TTF-1 and PLAGL-2 by real-time quantitative polymerase chain reaction.Independent sample t-test was used for statistical analysis.Results The average random blood glucose level in GDM group was significantly higher than that in control group [(26.8± 2.8) vs (4.9± 0.5) mmol/L,t=-34.05,P=0.00].The average weight of fetal rats in GDM group was higher than that in control group [(5.6±0.6) vs (5.2±0.5) g,t=-1.97,P=0.03].Alveolar number (10.1 ± 1.6 vs 12.1 ± 1.3) and alveolar area [(986.9 ± 5.5) vs (1 257.3± 5.0) μ m2] in GDM group was less than that in control group (t=9.84 and 27.53,both P ＜ 0.05).Alveolar septum [(11.5±6.2) vs (9.9±4.3) μm] in GDM group was higher than that in control group (t=-2.17,P ＜ 0.05).Microvillus in type] cells were short and the number of lamellar bodies was significantly decreased in GDM group.SP-B,SP-C,TTF-1 and PLAGL-2 proteins were distributed in the cytoplasm in granular form.The average value of absorbance of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 1.15±0.12,1.23±0.06,0.87±0.21 and 1.21 ±0.18 respectively;and that in control group was 1.22±0.05,1.31 ±0.14,1.12±0.09 and 1.33 ±0.07 respectively.The value in GDM group was lower than that in control group (t=2.40,2.35,4.89,and 2.77 respectively,all P ＜ 0.01).The expression level of SP-B,SP-C,TTF-1 and PLAGL-2 proteins in GDM group was 0.57± 0.09,0.45±0.03,1.50±0.04 and 1.11 ±0.04 respectively;and that in control group was 0.81 ±0.03,0.66±0.04,1.69±0.05 and 1.46±0.07 respectively.The value in GDM group was lower than that in control group (t=1 1.77,11.09,8.80 and 13.37,respectively,all P ＜ 0.01).The mRNA level of SP-B,SP-C,TTF-1 and PLAGL-2 in GDM group was 0.60±0.04,0.79±0.04,0.81 ±0.03 and 0.79±0.05 respectively;and that in control group was 1.06±0.19,1.03±0.24,1.03±0.18 and 1.02±0.19 respectively.The value in GDM group was lower than that in control group (t=6.80,2.98,3.54 and 3.54 respectively,all P ＜ 0.01).Conclusions The protein expression level of SP-B and SP-C in fetal lungs of GDM rats decreases obviously,possibly because of the down-regulation of the gene expression of TTF-1 and/or PLAGL-2.The pathological changes in fetal lungs of GDM rats might be associated with the descending level of SP-B and SP-C protein.

Effects of antenatal dexamethasone and ambroxol on expression of surfactant protein-B mRNA and ;thyroid transcription factor-1 in pulmonary tissue of premature rats / 中华围产医学杂志

To explore the effects and mechanism of dexamethasone and ambroxol on expression of surfactant protein (SP)-B mRNA and thyroid transcription factor (TTF)-1 in premature rat lung. Methods Sixteen pregnant Sprague-Dawley rats were randomly divided into four equal groups: two doses of dexamethasone (0.2 mg/kg injected intramuscularly on Day 17 and 18 of pregnancy respectively);single dose of dexamethasone (0.2 mg/kg injected intramuscularly on Day 18 of pregnancy);ambroxol group (100 mg/kg injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively); and control group (normal saline injected intraperitoneally on Day 16, 17 and 18 of pregnancy respectively). There were four pregnant rats in each group. All of the fetal rats were taken out on Day 19 of pregnancy as the preterm birth model, and 20 fetal rats from each group were randomly selected. The ratio of body weight to fetal lung weight of newborn rats was calculated. Changes in lung morphology were observed under light microscopy and the ratio of alveoli surface area to alveolar septae surface area was calculated. Expression of TTF-1 protein was determined by immunohistochemistry. Expression of SP-B mRNA was detected by reverse transcriptase-polymerase chain reaction. One-way analysis of variance, Student-Newman-Keuls method and Pearson correlation analysis were applied as statistical methods. Results (1) The ratio of body weight to fetal lung weight was (6.5±0.6), (7.9±0.8), (9.5±0.8) and (9.5±0.9) mg/g in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=67.69,P<0.01). The ratio of two doses and one dose of dexamethasone group was lower than that of control group (q=17.143 and 9.143, all P<0.01) and ambroxol group (q=17.143 and 9.143, all P<0.01). The ratio of two doses dexamethasone group was lower than that of one dose dexamethasone group (q=8.000, P<0.01). (2) The ratio of alveoli surface area to alveolar septae surface area was 2.19±0.15, 1.70±0.18, 1.67±0.13 and 1.08±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=190.85, P<0.01). The ratio of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of the control group (q=33.639, 18.788 and 17.879, all P<0.01). The ratio of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=14.848, P<0.01). (3) Expression of TTF-1 protein was 0.311±0.018, 0.224±0.019, 0.196±0.013 and 0.191±0.018 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=211.69,P<0.01). TTF-1 protein expression of two doses and one dose of dexamethasone group were higher than that of control group (q=30.000 and 8.250, all P<0.01) and ambroxol group (q=28.750 and 7.000, all P<0.01). TTF-1 protein expression of two doses dexamethasone group was higher than that of one dose dexamethasone group (q=21.750, P<0.01). (4) Expression of SP-B mRNA was 1.25±0.13, 1.15±0.12, 1.10±0.10 and 1.01±0.12 in two doses of dexamethasone group, one dose of dexamethasone group, ambroxol group and control group respectively (F=14.48, P<0.01). SP-B mRNA expression of two doses of dexamethasone group, one dose of dexamethasone group and ambroxol group were higher than that of control group (q=9.231, 5.385 and 3.462, all P<0.01). SP-B mRNA expression of two doses of dexamethasone group was higher than that of ambroxol group (q=5.769, P<0.01) and one dose of dexamethasone group (q=3.846, P<0.01). (5)TTF-1 expression in two doses of dexamethasone group, one dose of dexamethasone group and control groups was positively correlated with SP-B mRNA expression (r=0.512, 0.597 and 0.449, respectively, all P<0.05). Conclusions Ambroxol can accelerate the maturation of fetal lung with minimal adverse effects on fetal lung weight. Ambroxol might be an alternative to dexamethasone to prevent neonatal respiratory distress syndrome.

Clinical and radiological features in patients with DYT6 dystonia / 中华神经科杂志

Objective To summarize the clinical and radiological features of DYT6 dystonia with mutations based on the data of our patient cohort as well as the report by others.Methods Clinical data of the 11 patients with DYT6 dystonia in Peking Union Medical College Hospital from June 2009 to May 2012 were retrospectively reviewed and analyzed.Clinical data included gender,onset age,initiative symptom of onset,the sites of involvemet,family history,etc.All patients were examined for brain MRI scan,6 patients were examined for DTI.Results Of the eleven gene-confirmed DYT6 dystonia patients,7 were male and 4 were female,with an onset-age ranged from 5 years to 36 years,the mean age of onset was 19.4years.Eight patients had a family history.There were 10 patients with early onset dystonia and only 1 patient with late onset dystonia.The most common site of onset was the neck (7/11),and the next was the right arm,1-5 body areas were affected at the time of neurological assessment,the average amount was 2.8,and the most frequently affected anatomical site was the neck (10/11),next came lower face,jaw and tongue.Among all the patients,6 patients presented with segmental dystonia,4 patients presented with focal dystonia,only 1 patient presented with generalized dystonia.All the patients with thanatos-associated protein domain-containing apoptosis-associated protein (THAP) domain affected had a family history,but the patients with the same mutant gene varied with clinical manifestation.Only 1 patients with non-THAP domain affected had a family history,but in most families,there were adult asymptomatic mutant gene carriers.Mutations within the THAP domain were associated with an earlier age of onset than non-THAP domain (17.3 and 21.8 years old).Routine MRI of all patients were normal and DTI of 6 patients showed that fractional anisotropy values in the bilateral sensorimotor area in DYT6 dystonia were reduced.A detailed description of a patient with TOR1A and THAP1 gene mutations was given.Conclusions Early onset dystonia is the main manifestation in patients with DYT6 dystonia in China.The most common site of onset is the neck,and the next is the right arm.The most frequently affected anatomical site is the neck,next come lower face,jaw and tongue.Laryngeal dystonia is absent.The patients with same mutant gene show high heterogeneity in the clinical manifestations,mutations within the THAP domain of THAP1 tend to manifest at an earlier age and higher penetration than mutations localized to non-THAP domain.Reduction of fractional anisotropy values indicates that the axonal integrity and coherence in the region of sensorimotor area is damaged in DYT6 dystonia.

Detection of NPM1 gene mutation in acute myeloid leukemia / 中华检验医学杂志

Objective To analyze the frequency of NPM1 mutation in de novo acute myeloid leukemia (AML) patients and the relationship between NPM1 mutation and chromosome alterations,as well as FAB subgroups,and to analyze the mutation type.MethodsA total of 99 de novo AML patients from 2004 to 2010 in China-Japan Friendship Hospital were studied.Genomic DNA was amplified by polymerase chain reaction (PCR),denaturing polyacrylamide gel electrophoresis (PAGE) and capillary electrophoresis were used to detect the mutation of NPM1 gene in 99 AML patients,and karyotyping was performed in 72 AML patients by G banding techniques.DNA sequences analysis of NPM1 mutation was performed on 10 patients.Chi-square test was used to compare the frequencies of NPM1 mutation among the different subgroups,and McNemar's test was used to compare the different rates between denaturing PAGE and capillary electrophoresis.ResultsThe frequencies of NPM1 mutations were detected in 15％ (15/99) of AML patients with capillary electrophoresis and 11％ (11/99 ) with denaturing PAGE(x2 =2.25,P ＞0.05 ).The NPM1 was at different rates in M2(27％,8/30),M5(32％,6/19),M6( 13％,1/8),respectively (x2 =1.06,P ＞ 0.05 ),and not detected in the other subgroups.NPM1 mutation in patients with normal karyotype(26％ ) was more prevalent than patients with abnormal karyotype (4％) (x2 =5.61,P ＜ 0.05)All of the 10 patients were of A type ( c.860_863dupTCTG).The C-terminal portion of the NPM protein by replacing the last seven amino acids(WQWRKSL) with 11 residues (CLAVEEVSLRK).Two intronic deletions were novel,one case was IVS10-18_-15delCTTT,the other was IVS10-17_-15delTTT.Conclusions NPM1 mutations represents a common genetic abnormality in AML patients,and NPM1 mutation in patients with normal karyotype is higher than patients with abnormal karyotype.Two new intronic deletion mutations are identified.

Effect of XPG down-regulation gene expression towards the proliferation of epithelial ovarian cancer cells and its chemosensitivity to platinum / 中华妇产科杂志

ObjectiveTo investigate the effect of XPG down-regulation gene expression towards the proliferation of epithelial ovarian cancer cells and its chemosensitivity to platinum.Methods The small interference RNA ( siRNA ) -XPG fragments were designed and tranfected into SKOV3/DDP cell lines by lipofectamine transiently for choosing the best siRNA-XPG fragment to silence XPG gene expression.The pGPU6/GFP/Neo vector was used to construct the siRNA-XPG vectors,which was transfected into SKOV3/DDP cell line with expression of XPG gene.Real-time PCR and western blot were employed to confirm the silencing efficacy of siRNA-XPG.The growth curve of cells,cell cycle,the drug-resistance index of cells and intracellular drug concentration were measured by 4-methyl-thiazolyl-tetrazolium (MTT),flow cytometer (FCM) and high performance liguid chromatograph respectively.Results( 1 ) Real-time PCR results showed that XPG mRNA expression copy number in SKOV3/DDP tranfected with siRNA-XPG-733 fragment was 1.050 ± 0.023,which was significantly lower than that in SKOV3/DDP tranfected with other siRNA-XPG fragments(P ＜ 0.05,respectively),and was chosed to construct the siRNA-XPG vectors.The XPG mRNA expression was down-regulated in short hairpin RNA (shRNA)-XPG-733-SKOV3/DDP cell lines that confirmed by western blot.( 2 ) The growth curve showed that growth velocity of shRNA-XPG-733-SKOV3/DDP cell lines was lower than that of shRNA-GAPDH and shRNA-NC cell lines( P ＜ 0.05,respectively).The results of FCM also showed that 34.0％ of cells in shRNA-XPG-733-SKOV3/DDP cell lines were in S + G2/M phase,while only 58.7％ and 51.3％ in shRNA-GAPDH and shRNA-NC cell lines respectively ( P ＜ 0.05,respectively).( 3 ) The drug-resistance index of shRNA-XPG-733-SKOV3/DDP cell lines [ 50％ inhibiting concertration( IC50 ):( 13.79 ± 0.06) μg/ml ] was lower than that in shRNA-GAPDH and shRNANC cell lines [ IC50:( 27.84 ± 0.34 ) μg/ml and ( 28.32 ± 0.42 ) μg/ml,respectively ] statistically significant (P ＜ 0.05,respectively) ; but there was not statistically significant difference in intracellular drug concentration between shRNA-XPG-733-SKOV3/DDP cell lines [ (0.026 ± 0.005 ) μg/ml ] and shRNAGAPDH [ (0.024 ± 0.003 ) μg/ml ] and shRNA-NC cell lines [ ( 0.025 ± 0.007 ) μg/ml ] after treated by cisplatin in vitro ( P ＞ 0.05,respectively ).Conclusion The down-regulating of XPG gene resulted in slowing growth velocity and descending the drug-resistance index of shRNA-XPG-733-SKOV3/DDP cell lines,which may be related with descending in capability of DNA excision repair in cells.

Study on expressions of WWOX gene and p73 gene in acute lymphocytic leukemia patients / 中华检验医学杂志

Objective To investigate the clinical significance and mechanism of WW domain containing oxidoreductase (WWOX) gene and p73 gene abnormal expression in acute lymphocytic leukemia (ALL).MethodsCase-control study was used in the research.Forty-eight cases of bone marrows from ALL patients were collected,including 32 cases newly diagnosed,11 cases with complete remission and 5 case with relapse.Thirty-one cases of bone marrows from non-leukemia patients were used as control group.All the samples were collected from First Affiliated Hospital of Guangxi Medical University from July 2010 to July 2011.The doctors punctured patients' bone marrows 3 milliliters from the left of posterior superior iliac spine.Samples were bottled up with EDTA anti-coagulation tube.1 milliliter bone marrow was used to extract genome RNA with purity from 1.8 to 2.0.And then,the level of WWOX and p73 gene transcripts were tested immediately using reverse transcriptase-polymerase chain reaction (RT-PCR).Meanwhile genome DNA was also extracted from the other 2 milliliter bone marrow with purity from 1.7 to 1.9,which was used to detect the promoter methylation of WWOX gene and the first exon methylation of p73 gene by methylation PCR (MS-PCP).x2 test and Fisher's exact test were used to compare tbe methylation status of WWOX and p73 gene.Results In 31 controls,expression of WWOX and p73 gene mRNA was 94.00％.The total expression frequency of WWOX gene mRNA in 48 ALL samples was 48.00％ (23/48),much lower than control (x2 =17.434,P =0.000 ).There was significant difference (x2 =10.471,P =0.001 ) between newly diagnosed cases 34.38％ ( 11/32),complcte remission cases (90.91％,10/11 ) and control.The total expression frequency of p73 gene mRNA in 48 ALL samples was 56.00％ (27/48),much lower than control (x2 =12.697,P =0.000).There was significant difference (P =0.012 ) between newly diagnosed cases 43.75％(14/32) and complete remission cases 90.91％(10/11).It was unmethylation in 31 controls.The total methylation frequency of WWOX gene promoter region in 48 ALL samples was 44.00％(21/48),much lower than control (x2 =18.473,P =0.000).There was significant difference (P =0.012) between newly diagnosed cases 56.25％ (18/32),complete remission cases 9.09％ (1/11 ) and control.The total methylation frequency of p73 gene the first exon region in 48 ALL samples was 35.00％(17/48),much lower than control (x2 =13.990,P =0.000).There was significant difference (P =0.033) between newly diagnosed cases 46.88％ (15/32),complete remission cases 9.09％ ( 1/11 ) and control.There was a negative correlation between the expression of WWOX gene mRNA and its methylation status(r =- 0.678,P =0.000),the same as p73 gene ( r =- 0.577,P =0.000).ConclusionsThe abnormal methylation of WWOX and p73 gene may be the major mechanism of gene silence in ALL,which leads to no expression of WWOX mRNA or p73 mRNA.And the abnormal methylation of WWOX and p73 gene may be relevant with the process of occurrence and development in ALL.It may be an effective and significant to detect methylation status of WWOX gene and p73 gene for the diagnosis and treatment of ALL patients.(Chin J Lab Med,2012,35:820-825)

Study on the expression and significance of nucleolin in osteosarcoma / 中国医师杂志

ObjectiveTo investigate the expression of nucleolin in osteosarcoma and its clinicalsignificance.MethodsThe expression of nucleolin was evaluated by immunohistochemistry in paraffin-em bedded tissues from 35 cases of osteosarcoma and 15 cases of osteochondroma.ResultsAmong the osteo sarcoma cases, 8.6％ were negative for nucleolin, 20％ weakly positive, 34.3％ moderate positive, 37.1％ strong positive.The positive rate of nucleolin in osteochondroma was 40％, 46.7％, 13.3％, 0％, respectively.The nucleolin expression showed significant difference in the osteosarcoma and osteochondroma(P ＜0.01).Among the conventional ostcosarcoma cases, 6.9％ were negative for nucleolin, 13.8％ weakly positive, 34.5％ moderate positive, 44.8％ strong positive.The positive rate of nucleolin in paros teal osteochondroma was 16.7％, 50％, 33.3％, 0％, respectively.The nucleolin expression showed re markable correlation with pathological and genetical classification (P ＜ 0.05).The positive rate of nucleo 1in in osteoblastic osteochondroma was 5.9％, 5.9％, 29.4％, 58.8％, while in chondroblastic osteochon droma 0％, 33.3％, 33.3％, 33.3％ and 16.7％, 16.7％, 50％, 16.7％ in fibroblastic osteochondro ma, respectively.The positive expressions of nucleolin were almost the same in each subtype of osteosarco ma (P ＞ 0.05).ConclusionsThe nucleolin expression had correlation with property of tumors in bone, and the positive expressions of nucleolin had correlation with pathological and genetic classification of osteo

Expression of growth arrest and DNA damage-inducible 45α and p38 mitogen-activated protein kinase in placentas of women complicated with preeclampsia / 中华围产医学杂志

Objective To evaluate the expression of Gadd45α and p38 MAPK in placentas and the correlations of Gadd45α protein and serum soluble vascular endothelial growth factor receptor-1 (sFlt-1) and soluble endoglin (sEng) in preeclampsia(PE). Methods Fifty-four pregnant women who delivered from September 2009 to March 2010 in the First Affiliated Hospital of Chongqing Medical University were chosen as the subjects. They were classified into mild preeclampsia group (n=20),severe preeclampsia group (n=16) and the control group (normal pregnant women underwent elective cesarean sections at term without labor and perinatal complications, n=18). Western blot and immunohistochemistry were employed to determine the expression and localization of Gadd45α and p-p38 MAPK protein respectively. Gadd45α mRNA level was determined by quantitative real-time PCR. The levels of seum sFlt-1 and sEng were measured by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and LSD-t test were applied for statistical analysis. Results (1)Immunohistochemistry identified that the positive stained cells were mostly located in trophoblast cells in normotensive placentas, whereas in preeclamptic placentas Gadd45α protein and p-p38 MAPK protein were detected in trophoblast and endothelial cells, as well as a few stromal cells at increased levels.(2)The mRNA levels of Gadd45α was significantly elevated in mild and severe preeclampsia groups compared to the control group (2.10±0.11 and 3.33±0.13 vs 1.01±0.18, P＜0.05), and Gadd45α mRNA level in severe group was significantly higher than in mild group (P＜0.05).(3)The data of Western blot revealed that the Gadd45α protein levels in each group were 0.22±0.11, 0.65±0.15 and 1.34±0.17, respectively, with significant differences between each group(P＜0.05). The p-p38 MAPK protein levels in each group were 0.32±0.08, 0.72±0.12 and 1.45±0.21, respectively, with significant differences between each group (P＜0.05). p38 MAPK protein levels in the total groups showed no difference(P＞0.05).(4)Compared with the control group, sFlt-1 and sEng concentrations in maternal circulation were significantly increasing in mild and severe preeclampsia groups, and concentrations in severe group were significantly higher than those in mild group (P＜0.05).(5) There were positive correlations between Gadd45α protein levels and the concentrations of serum sFlt-1 and sEng in each group( r=0.88 and 0.87, respectively all P＜0.05). Conclusions Upregulation of Gadd45α in preeclampsia placentas may play an important role in the pathogenesis of preeclampsia. It may induce the increased maternal serum levels of sFlt-l and sEng by activating p38 MAPK signaling pathway, leading to deficient cytotrophoblastic invasion and abnormal placental vascular reconstruction during pregnancy.