Objective:To investigate the effects of pregnancy and lactation nonylphenol (NP) exposure on the balance of Treg/Th17 cells in the brain of offspring mice and the related mechanisms.Methods:Thirty pregnant C57BL/6 mice were randomly divided into three groups: control group (drinking distilled water), and NP-treated groups (drinking 0.2 μg/ml or 2.0 μg/ml NP water solution). ELISA kit was used to analyze the levels of TNF-α, IFN-γ and IL-17, flow cytometry was used to analyze the frequency of Treg and Th17 cells in spleen, quantitative RT-PCR was used to analyze the RORγt, Foxp3 mRNA, Western blot was used to analyze the protein expression of RORγt, Foxp3 and PI3K/Akt/mTOR signal pathway, and immunofluorescence was used to analyze the expression of Iba1 in the brain tissue of offspring mice.Results:Compared with the control group, NP exposure increased the serum levels of IL-17 and TNF-α in male offspring mice ( P<0.05), and decreased the levels of IFN-γ( P<0.05). Flow cytometry analysis showed that the percentage of Th17 cells in the spleen of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly higher than that of the control group, while the percentage of Tregs cells was lower. Compared with the control group, the expression levels of Foxp3 proteins in the brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) was significantly lower, accompanied by a dramatic increase in RORγt protein levels ( P<0.05). Similar mRNA expression was also observed in qRT-PCR analysis. The protein expression levels of mTOR (p-mTOR) and its upstream related regulators[PI3K, p-Akt (Ser473), p-Akt (Thr308)] in the brain of male offspring mice increased gradually during the period of exposure to NP( P<0.05). Immunofluorescence analysis showed that compared with the control group, the number of Iba1 positive cells in brain tissue of male offspring mice exposed to NP (0.2 μg/ml or 2.0 μg/ml) increased significantly ( P<0.05). Conclusions:Maternal exposure to NP during pregnancy and lactation may affect the development/function of neurons in offspring through neuroimmune axis, and increase the risk of neurodevelopmental disorders in offspring.

ObjectiveTo investigate the role of T helper (Th) 22 and Th17 cells in the pathogenesis of severe preeclampsia.MethodsThirty women with severe preeclampsia who delivered in the Third Affiliated Hospital of Zhengzhou University from October 2014 to February 2016 were enrolled in the study. Thirty healthy pregnant women matched for age and gestational weeks were recruited as the control group. The frequencies of Th22 and Th17 cells in peripheral whole blood were determined by flow cytometry. The concentrations of interleukin (IL)-22 and IL-17A in plasma were detected by enzyme-linked immunosorbent assay. Independent two samplest-test, non-parametric test and Spearman correlation analysis were used for statistical analysis.ResultsThe percentage of Th22 and Th17 cells in the severe preeclampsia group were significantly higher than those in the control group, respectively[Th22 cells: 0.59% (0.39%-1.13%) vs 0.40%(0.23%-0.57%),Z=2.530,P=0.010; Th17 cells: 3.24% (3.02%-3.97%) vs 1.87% (1.53%-2.64%),Z=5.046, P=0.000]. So were the plasma levels of IL-22 and IL-17A[IL-22: 285.72 (247.63-306.69) vs 233.85 (184.92-258.38) pg/ml,Z=4.341,P=0.001; IL-17A: 27.53 (23.84-32.78) vs 17.36 (15.58-19.13) pg/ml,Z=4.924, P=0.000]. There was a positive correlation between circulating Th22 and Th17 cells in the severe preeclampsia group (r=0.534,P=0.015), while no correlation was found in the control group (r=0.345,P=0.136). Positive correlation was found in plasma level of IL-22 with Th22 cells (r=0.600,P=0.005), but not with Th17 cells (r=0.398,P=0.082) in the severe preeclampsia group.ConclusionsIncreased Th22 cells and high IL-22 concentrations in the peripheral blood of severe preeclampsia patients may indicate a self-defense mechanism in the maternal body. Th22 cells and Th17 cells may interact with each other.

Objective To investigate the expression changes and clinical significance of microRNA-155(miR-155) and chemokine receptor 4(CXCR4) in placental tissue from the patients with preeclampsia(PE).Methods Thirty pregnant women with severe PE(sPE) in the Third Affiliated Hospital of Zhengzhou University from December 2015 to February 2016 served as the sPE group,and contemporaneous 30 healthy pregnant women undergoing cesarean section due to the social factors served as the healthy control group(N).The real-time fluorescent quantitative PCR(RT-PCR) was used to detect the expression of miR-155 and CXCR4 mRNA in placental tissue and the relationship between miR-155 and CXCR4 levels was analyzed.The immunohistochemistry SABC methods were used to detect the expression of CXCR4 protein in villous cytotrophoblast(VCT) tissue microarray(TMA,42 cases in the normal control group 1,56 cases in the PE group) and extravillous cytotrophoblast(EVCT) TMA(29 cases in the normal control group 2,47 cases in the PE group) constructed by the same research group.Results (1) There was no statistically significant difference in the age,gestational age and pre-pregnancy body mass index(BMI) between the two groups(P>0.05).The differences of blood pressure between the two groups were statistically significant(P<0.05).The neonatal birthweight in the sPE group was significantly lower than that in the control group with statistically significant differences(P<0.05).The urine protein in the sPE group was significantly higher than that in the control group with statistically significant differences(P<0.05).(2)In the placental VCTand EVCT TMA,the age had no statistical difference between the two groups(P>0.05).The gestational weeks of the PE group were earlier than those in the N group 1,the systolic/diastolic blood pressure and urine protein were higher than those in the N group,the differences all were statistically significant(P<0.05),the neonatal birthweight was significantly lower than that in the N group with statistically significant differences(P<0.05).(3)The expression level of miR-155 mRNA in placental tissue in the sPE group was 1.53±0.92,which was significantly higher than 0.87±0.73 in the control group,the difference between the two groups was statistically significant(P<0.05).(4) The expression level of CXCR4 mRNA in the N group was 1.51±1.85,which in the sPE group was 0.54±0.38,the difference between the two groups was statistically significant(P<0.05).(5) In the sPE group,the miR-155 level and CXCR4 level in placntal tissue had a significant correlation(r=-0.773,P<0.05).(6) CXCR4 protein was expressed in VCT and EVCT TMA;the CXCR4 positive expression rate of the PE group in VCT TMA was 48.21%(27/56),which in the sPE group was 47.92%(23/48) and which in the early onset PE group was 53.66%(22/41),which all were significantly lower than 83.33%(35/42)in the normal control group,the differences all were statistically significant(P<0.05).(7)The positive expression rate of CXCR4 in the PE group in placent EVCT TMA was 48.94%(23/47),which in the sPE group was 50.00%(22/44) and which in the early PE onset group was 52.63%(20/38),which all significantly lower than 79.31%(23/29) in the normal control group,the differences all were statistically significant(P<0.05).Conclusion The level of placental tissue miR-155 is increased in the patients with sPE,while the level of CXCR4 is decreases obviously,both have a negative correlation,which may be associated with the pathogenesis of PE.

Objective:To investigate the effect on the CXCR4/PI3K/AKT pathway after the transfection of miR-155 mimics and miR-155 inhibitor combined with the research on the ability of invasion and migration of human chorionic JEG-3 trophoblast cells. Methods:Chemically synthesized miR-155 mimics and miR-155 inhibitor were transfected into JEG-3 cells. The effect on the ability of invasion and migration were analyzed by Transwell migration assay and Wound healing assay. The expression of CXCR4 mRNA was detected by Real-time PCR. The expression of CXCR4 and p-AKT protein were detected by Western blot. Results: Transfection with miR-155 mimics significantly down-regulated the expression of CXCR4 as compared with the control group(P<0. 05);JEG-3 cells transfected miR-155 mimics had lower levels of migration and invasion capacity than cells in the control group(P<0. 05). However, transfection with miR-155 inhibitor significantly up-regulated the expression of CXCR4 as compared with the control group(P<0. 05);JEG-3 cells transfected miR-155 inhibitor had higher levels of migration and invasion capacity than cells in the control group ( P<0.05).Addition,the expression of p-AKT protein of JEG-3 cells was down-regulated after transfected miR-155 mimics,and the expression of p-AKT protein of JEG-3 cells was up-regulated after transfected miR-155 inhibitor. Conclusion:miR-155 may inhibits the invasion and migration of trophoblast cells by regulating CXCR4/PI3K/AKT pathway contributing to the development of preeclampsia.

Objective To investigate the effects of stromal cell-derived factor 1 (SDF-1) and an CXC chemokine receptor 4 (CXCR4) antagonist (AMD3100) on the invasion and migration capabilities of the huaman choriocarcinoma cell line JAR for further elucidating the role of SDF-1/CXCR4 axis in the pathogenesis of preeclampsia.Methods JAR cells were divided into four groups: SDF-1 group (treated with 50 ng/ml of SDF-1),SDF-1+AM3100 mixed group (first treated with 100 ng/ml of AMD3100 for 2 hours and then treated with 50 ng/ml of SDF-1),AMD3100 group (treated with 100 ng/ml of AMD3100) and blank control group (without any treatment).RT-PCR was performed to detect the expression of CXCR4 at mRNA level in JAR cells.Western blot assay was used to measure the expression of CXCR4 and p-AKT at protein level.MTT assay was used to analyze the effects of different concentrations of SDF-1 (10,30,50 and 100 ng/ml) on the proliferation of JAR cells at different time points (0,24,48,72 h).Transwell invasion assay and wound-healing assay were used to test the changes in invasion and migration capabilities of JAR cells after different treatments.Results (1) Results of the RT-PCR showed that the expression of CXCR4 at mRNA level in JAR cells was increased in the SDF-1 group (1.839±0.083) as compared with that in the blank control group (1.372±0.086),AMD3100 group (0.694±0.045) or SDF-1+AM3100 mixed group (0.703±0.093).Moreover,the differences between the SDF-1 group and the other three groups were statistically significant (F=30.67,P<0.05).Compared with the blank control group,the expression of CXCR4 at mRNA level in JAR cells was decreased in the AMD3100 group (P<0.01).(2) Results of the Western blot assay showed that the expression of CXCR4 and p-AKT at protein level in JAR cells were enhanced in the SDF-1 group as compared with that in the blank control group,AMD3100 group or SDF-1+AM3100 mixed group.Compared with the blank control group,the expression of CXCR4 and p-AKT at protein level in JAR cells were inhibited in the AMD3100 group.(3) Results of the MTT assay showed that SDF-1,especially at the concentration of 50 ng/ml,could enhance the proliferation of JAR cells (P<0.05) and its best effect on proliferation was seen at 48 h.(4) Results of the Transwell invasion assay showed that the number of transmembrane cells in the SDF-1 group (70.49±2.42) was more than that in the blank control group (54.36±2.26),AMD3100 group (21.68±8.31),or SDF-1+AMD3100 mixed group (28.18±4.61).The differences between the SDF-1 group and the other three groups were statistically significant (F=116.26,P<0.01).Compared with the blank control group,the number of transmembrane cells was reduced in the AMD3100 group (P<0.05).(5) Results of the wound-healing assay showed that the relative migration distance was increased in the SDF-1 group (1.162±0.034) as compared with that in the blank control group (0.823±0.101),AMD3100 group (0.160±0.047),or SDF-1+AMD3100 mixed group (0.183±0.064).The differences between the SDF-1 group and the other three groups were statistically significant (F=30.500,P<0.05).Compared with the blank control group,the relative migration distance was decreased in the AMD3100 group (P<0.01).Conclusion The invasion and migration of huaman choriocarcinoma JAR cells can be enhanced by SDF-1,but inhibited by AMD3100.This study indicates that the blocked biological axis of SDF-1/CXCR4 may play an important role in the pathogenesis of preeclampsia through inducing abnormal activation of PI3K/AKT pathway,which results in inhibited invasion and migration of trophoblast cells and placenta abnormality.

Objective To observe the effect of Triclosan( TCS) exposure on Caenorhabditis elegans( c. ele-gans) F1 generation of locomotory behavior, brood size, and generation time. Methods The trial included a control group and 4 TCS treatment groups with different doses (100 nmol/L,1 μmol/L,10μmol/L,20μmol/L),the exposure time being 24 hours,the effect of c. elegans′head thrashes,body bending frequency,the brood size and generation time was observed. Results (1) The control group exposed to 100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,their head thrash frequency of c. elegans F1 was(109. 40±8. 61) times/min,(84. 70±7. 82) times/min,(76. 35±7. 44) times/min,(74. 74±5. 93)times/min,(71. 95±4. 19)times/min,respectively,the head thrash ability of c. elegans was significantly inhibited(F=62. 245,P<0. 01). (2) When the control group was exposed to 100 nmol/L,1 μmol/L,10μmol/L,20 μmol/LTCS,the frequency of c.elegans F1 body bent was (19.94±2.46)times/20 s,(15.13±1.99) times/20 s,(14.63±2.31)times/20 s,(14.69±1.96)times/20 s,(12.00±1.86)times/20 s,respectively,and the comparative differences between groups were statistically significant(F=25. 636,P<0. 01). (3) When the control group was exposed to 0,100 nmol/L,1 μmol/L,10 μmol/L,20 μmol/L TCS,the body sizes of the c. elegans F1 generation was (286.83±6.01)articles,(273.33±6.41)articles,(214.17±7.25)articles,(173.67±9.20)articles, (118. 50 ± 6. 98 ) articles, respectively, the brood size of the C. elegans F1 generation exposed to 100 nmol/L, 1μmol/L,10 μmol/L,20 μmol/L TCS levels,were reduced by 4. 71%,25. 60%,39. 45%,58. 67%,the ge-neration time of the c. elegans′F1 generation was shortened by 2. 14%-5. 38% in the TCS treatment groups compared with the control group(F=27. 520,P<0. 01). Conclusions After c. elegans exposure to TCS,locomotory behavior can be severe-ly affected,reproductive damage causes a decline in the number of brood size,and the speeding-up of the breeding rate is related to the concentration of TCS concentration-response.

Objective To investigate the expression and role of regulatory plasma cells in gravidas with systemic lupus erythematosus ( SLE) . Methods Gravidas with SLE were enrolled in Henan Provincial People's Hospital from April 2013 to April 2018. They were divided into three groups including pregnancy control group, SLE stable group and SLE deterioration group. The ratio of CD3-LAG-3+CD138high regulatory plasma cells was detected by flow cytometry. The concentrations of soluble human leukocyte antigen-G ( sHLA-G) and anti-nuclear antibody Ig were detected by ELISA. Lymphocytes in peripheral blood of SLE deterioration group were isolated, and then cultured in RPMI1640 medium containing 10％ fetal bovine ser-um and stimulated with HLA-G. Results Flow cytometry showed that the proportion of regulatory plasma cells in SLE stable group was (2. 483±0. 1318)％ and that in SLE deteriorating group was (1. 662± 0. 1304)％. There was a significant difference between the two groups (t=4. 431, P=0. 0013). The con-centrations of sHLA-G in SLE stable group and SLE deteriorating group were (36. 50±3. 510) ng/ml and (16. 50±2. 405) ng/ml, and the difference between the two groups was statistically significant (t=4. 701, P=0. 0008). Correlation analysis showed that the concentration of sHLA-G was positively correlated with the proportion of regulatory plasma cells (r=0. 7471, P=0. 0009). The results of in vitro experiment showed that the proportions of B cells and regulatory plasma cells were ( 7. 573 ± 0. 6539 )％ and ( 1. 593 ± 0.1879)％ in SLE deterioration group and (3. 732±0. 7178)％ and (2. 503±0. 2921)％ in HLA-G group with statistical differences between the two groups (t=3. 957, P=0. 0027;t=2. 620, P=0. 0256). Conclusions The proportion of regulatory plasma cells and the concentration of sHLA-G were significantly decreased in pregnant patients with SLE, which was closely related to disease severity. HLA-G played an important role in promoting the proliferation of regulatory plasma cells.

Mycoplasma pneumoniae pneumonia is an important component of community-acquired pneumonia, which can cause severe extrapulmonary complications of digestive, cardiovascular, blood, urinary and other systems.Accurate and effective biomarkers are significant for the diagnosis and treatment of Mycoplasma pneumoniae pneumonia.Recent studies have shown that new biomarkers such as IL-35, IL-17, neutrophil to lymphocyte ratio(NLR) and S100 protein are involved in the development of Mycoplasma pneumoniae pneumonia.In order to provide reference for the clinical diagnosis and treatment of Mycoplasma pneumoniae pneumonia, this paper reviews the progress of new biomarkers in Mycoplasma pneumoniae pneumonia.

BACKGROUND:Premature birth is a major global health problem associated with high mortality and morbidity.White matter injury is the most common brain injury in preterm infants.Salvia miltiorrhiza is a traditional herbal plant that is commonly used to treat cardiovascular and cerebrovascular diseases in Asian countries. OBJECTIVE:To investigate the therapeutic effect of Salvia miltiorrhiza on white matter injury in preterm infants. METHODS:Eighteen neonatal male Sprague-Dawley rats at 3-day gestational age were selected and randomized into normal group,white matter injury group,and Salvia miltiorrhiza group.Animal models of preterm white matter injury were established by permanent ligation of the right common carotid artery in the latter two groups.Rats in the Salvia miltiorrhiza group were given intraperitoneal injection of Salvia miltiorrhiza(5 mg/kg·d)for 7 consecutive days.Normal group and white matter injury group were given the same volume of PBS for intervention.On the 14th day after modeling,the rats were sacrificed.Brains were pathologically observed by hematoxylin-eosin staining under microscope,and the expression levels of myelin basic protein and CC1 in brain tissue were visualized using immunofluorescence.Furthermore,liquid chromatography-tandem mass spectrometry was used to analyze possible pathways for the action of Salvia miltiorrhiza. RESULTS AND CONCLUSION:In the white matter injury group,the structure of the corpus callosum was irregular and the cells appeared swollen and necrotic.In addition,induction of white matter injury resulted in significantly reduced myelin formation,with irregular and loosely arranged nerve fibers and significantly decreased myelin sheaths.Interestingly,white matter injury rats treated with Salvia miltiorrhiza had reduced cellular swelling,reduced lesions,and increased myelin sheaths.The expression of myelin basic protein was closely related to myelin formation,and CC1 was a marker of myelin oligodendrocytes.Salvia miltiorrhiza significantly up-regulated the expressions of myelin basic protein and CC1 in white matter injury rats(P＜0.000 1),indicating that Salvia miltiorrhiza alleviated white matter injury.Liquid chromatography-tandem mass spectrometry analysis showed that the therapeutic effect of Salvia miltiorrhiza in the rat model of white matter injury was closely related to the regulation of complement and coagulation cascades.To conclude,Salvia miltiorrhiza may be a potential therapeutic agent for treating preterm white matter injury.