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Viruses infect host cells by binding to receptors on thesurface of cells. Receptor is an important factor affecting host range and interspecific transmission. In December 2019, an outbreak of unexplained pneumonia occurred in Wuhan, Hubei province. The pathogen was a new coronavirus, named 2019 NovelCoronavirus (2019-nCoV) by WHO. Angiotensin-converting enzyme 2 (ACE2) was found to be the receptor of 2019-nCoV.This review provides a brief overview of human coronavirus receptors and their applications, with a view to providing references for the tracing, cross-species transmission, epidemiological analysis and antiviral and vaccine studies of 2019-nCoV.
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Objective To investigate the regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer (dNK) cells during early pregnancy and its association with the incidence of recurrent spontaneous abortion (RSA). Methods Decidua tissues were collected from normal pregnancies (n=16) and RSA cases (n=6) at 6-10 gestational weeks in the Department of Obstetrics and Gynecology of Peking University Third Hospital from October 2018 to May 2019. (1) Expressions of CLEC2D and CD161 in decidua from early pregnancy were detected using immunofluorescence. (2) Primary dNK cells were isolated from decidua from early pregnancy. dNK cells pre-treated with CD161 antibody (blocking CD161, B-CD161) were co-cultured with JEG-3 cells which were knocked-down by CLEC2D small interfering RNA (siCLEC2D), followed by killing capacity assessment of dNK cells by cytotoxicity assay and determination of expressions of related molecules by quantitive real-time polymerase chain reaction. (3) Western blot and flow cytometry were used to detect the expression of CLEC2D and CD161 in decidua tissues. Cytotoxicity assay was performed to analyze the killing capacity of dNK cells. T test was used for statistical analysis between normal and RSA cases. Results (1) CLEC2D was mainly expressed in extravillous trophoblast (EVT) cells and CD161 was mainly detected in dNK cells. CD161-positive dNK cells and CLEC2D-positive EVT cells were adjacently located in decidua tissues allowing their interaction. (2) Cytotoxicity assay suggested that CD161 blocking in dNK cells or CLEC2D knockdown in JEG-3 cells could enhance the cytotoxicity of dNK cells. The target cell lysis rates at the effector-target ratios of 40 ∶ 1, 20 ∶ 1, 10 ∶ 1 and 5 ∶ 1 in B-CD161 group were (59.12±4.56)%, (25.96±5.44)%, (13.60±8.94)% and (12.53±8.94)%, and in IgG control group were (20.01±1.96)%, (8.51±1.32)%, (3.24±0.75)% and (3.82±1.92)%, respectively. There were significant differences between the two groups at the effector-target ratios of 40∶1 (t=13.922, P<0.01) and 20∶1 (t=5.403 P<0.05), but not at 10∶1 or 5∶1 (P>0.05). The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5 ∶ 1 in si-CLEC2D group were (43.37±2.01)%, (32.99±2.08)%, (23.47±1.36)% and (11.48±0.37)%, and in the negative control (NC) group were (15.54±1.46)%, (13.84±1.68)%, (9.94±3.01) and (5.50±0.99)%, respectively. Differences between the two groups at all effector-target ratios were statistically significant (t=19.402, 12.400, 7.093 and 9.842, all P<0.01). Moreover, the expression of dNK killing-related factor granzyme B in the siCLEC2D group was higher than that in the NC group. (3) Compared with the normal pregnancy group, the RSA group showed decreased CD161 expression and increased killing capacity of dNK cells, but no significant difference in CLEC2D expression. Conclusions At early pregnancy, CLEC2D on EVT cells can interact with CD161 on dNK cells, which inhibits the cytotoxicity of dNK cells and induces immune tolerance at the fetal-maternal interface. Decreased expression of CD161 in decidua results in increased cytotoxicity of dNK cells, which may be one of the causes of immune rejection in RSA.
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Objective@#To investigate the regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer (dNK) cells during early pregnancy and its association with the incidence of recurrent spontaneous abortion (RSA).@*Methods@#Decidua tissues were collected from normal pregnancies (n=16) and RSA cases (n=6) at 6-10 gestational weeks in the Department of Obstetrics and Gynecology of Peking University Third Hospital from October 2018 to May 2019. (1) Expressions of CLEC2D and CD161 in decidua from early pregnancy were detected using immunofluorescence. (2) Primary dNK cells were isolated from decidua from early pregnancy. dNK cells pre-treated with CD161 antibody (blocking CD161, B-CD161) were co-cultured with JEG-3 cells which were knocked-down by CLEC2D small interfering RNA (siCLEC2D), followed by killing capacity assessment of dNK cells by cytotoxicity assay and determination of expressions of related molecules by quantitive real-time polymerase chain reaction. (3) Western blot and flow cytometry were used to detect the expression of CLEC2D and CD161 in decidua tissues. Cytotoxicity assay was performed to analyze the killing capacity of dNK cells. T test was used for statistical analysis between normal and RSA cases.@*Results@#(1) CLEC2D was mainly expressed in extravillous trophoblast (EVT) cells and CD161 was mainly detected in dNK cells. CD161-positive dNK cells and CLEC2D-positive EVT cells were adjacently located in decidua tissues allowing their interaction. (2) Cytotoxicity assay suggested that CD161 blocking in dNK cells or CLEC2D knockdown in JEG-3 cells could enhance the cytotoxicity of dNK cells. The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in B-CD161 group were (59.12±4.56)%, (25.96±5.44)%, (13.60±8.94)% and (12.53±8.94)%, and in IgG control group were (20.01±1.96)%, (8.51±1.32)%, (3.24±0.75)% and (3.82±1.92)%, respectively. There were significant differences between the two groups at the effector-target ratios of 40∶1 (t=13.922, P<0.01) and 20∶1 (t=5.403 P<0.05), but not at 10∶1 or 5∶1 (P>0.05). The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in si-CLEC2D group were (43.37±2.01)%, (32.99±2.08)%, (23.47±1.36)% and (11.48±0.37)%, and in the negative control (NC) group were (15.54±1.46)%, (13.84±1.68)%, (9.94±3.01) and (5.50±0.99)%, respectively. Differences between the two groups at all effector-target ratios were statistically significant (t=19.402, 12.400, 7.093 and 9.842, all P<0.01). Moreover, the expression of dNK killing-related factor granzyme B in the siCLEC2D group was higher than that in the NC group. (3) Compared with the normal pregnancy group, the RSA group showed decreased CD161 expression and increased killing capacity of dNK cells, but no significant difference in CLEC2D expression.@*Conclusions@#At early pregnancy, CLEC2D on EVT cells can interact with CD161 on dNK cells, which inhibits the cytotoxicity of dNK cells and induces immune tolerance at the fetal-maternal interface. Decreased expression of CD161 in decidua results in increased cytotoxicity of dNK cells, which may be one of the causes of immune rejection in RSA.
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Objective To evaluate the effect of pulsed radiofrequency (PRF) on spinal adenosine triphosphate (ATP)-P2X4-NLRP3 signaling pathway in rats with neuropathic pain.Methods Forty healthy clean-grade adult male Sprague-Dawley rats,aged 2-3 months,weighing 220-260 g,were divided into 4 groups (n =10 each) using a random number table method:sham operation group (group S),neuropathic pain group (group NP),sham PRF group (group SPRF) and PRF group.Neuropathic pain was induced by chronic constriction injury to the left sciatic nerve of anesthetized rats.Rats received PRF treatment on 7th day after establishing the model in group PRF.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before establishing the model (T0) and at 3,7,10,14,21 and 28 days after establishing the model (T1-6).The rats were then sacrificed and the spinal cord was removed for determination of P2X4 and NLRP3 expression (by Western blot) and interleukin-1beta (IL-1β),IL-2,IL-6 and tumor necrosis factor-alpha (TNF-α) contents (by enzymelinked immunosorbent assay).Results Compared with group S,the MWT and TWL were significantly decreased at T1-6,the expression of P2X4 and NLRP3 was up-regulated,and the contents of IL-1β,IL-2,IL-6 and TNF-α were increased in NP,SPRF and PRF groups (P＜0.05).Compared with group NP and group SPRF,the MWT and MWT were significantly increased at T3-6,the expression of P2X4 and NLRP3 was down-regulated,and the contents of IL-1 β,IL-2,IL-6 and TNF-α were decreased in group PRF (P＜0.05).Conclusion The mechanism by which PRF alleviates neuropathic pain is related to inhibiting ATP-P2X4-NLRP3 signaling pathway in rats.
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Objective To investigate the effect of regulating peroxisome proliferator-activated receptor γγ (PPAR γ) on soluble endoglin (sEng) expression in first-trimester trophoblasts via an in vitro study.Methods Chorionic villus were collected from 20 samples of first-trimester artificial abortion in Peking University First Hospital from July 1 st to 31 st,2016.Primary culture of trophoblast cells was performed.Trophoblast cells from each sample were divided into three groups,which were PPAR γ antagonist group,PPAR γ antagonist and PPAR γ agonist group,and control group.Supematant sEng level was detected in each group by enzyme linked immunosorbent assay (ELISA).Paired-sample t test was used for statistical analysis.Results Compared with the control group,trophoblast cells in the PPAR γ antagonist group grew slower and were reduced in number.No significant difference in growth or morphology of trophoblast cells was observed between the PPAR γγ antagonist and PPAR γγ agonist group and the control group.Supernatant sEng level was elevated in the PPAR γ antagonist group,but was not significantly changed in the PPAR γ antagonist and PPAR γ agonist group as compared with that in the control group [(124.1 23.8) vs (94.0± 12.7) pg/ml,t=-4.31,P＜0.05;(87.1 ± 10.6) vs (94.0± 12.7) pg/ml,t=1.62,P=0.12).Conclusions Suppression of PPAR γ promotes sEng expression in trophoblast cells and that can be reversed by PPAR γ agonist.
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Objective To investigate the expression of endothelial protein C receptor (EPCR) and its roles in plasma and placenta of patients with early onset severe preeclampsia. Methods Sixty cases of severe preeclampsia women who delivered in Xuzhou Maternity and Child Health Care Hospital from March 2014 to February 2016, were recruited, which included 30 cases with early onset severe preeclampsia (early onset group, gestational week <34 weeks ) and 30 patients with late onset severe preeclampsia (late onset group, gestational week ≥34 weeks). Thirty cases of healthy late pregnant women at the same period (gestational week≥34 weeks) were selected as control group. Immunohistochemistry SP method was applied to detect the expression of in EPCR placenta. Reverse transcription (RT)-PCR was used to detect the expression of EPCR mRNA in placenta. ELISA method was used to detect the levels of soluble EPCR (sEPCR)level in plasma of the pregnant women of the three groups. Results The expression of EPCR in placenta mainly distributed in the membrane and cytoplasm of placental syncytiotrophoblasts and vascular endothelial cells, a few in the cell nucleus. The expression of EPCR in early onset group(57%, 17/30)was significantly lower than that in late onset group (93%, 28/30; χ2=25.165,P=0.001). The expression of EPCR in late onset group had no significant difference from that in control group (97%, 29/30;χ2=0.540,P=0.910). The expression of EPCR mRNA in placenta of early onset group(0.40±0.07)was significantly lower than that in late onset group(0.91±0.06;t=-30.044,P=0.001), while there was no statistical difference of the expression of EPCR mRNA between the late onset group and the control group (0.92±0.07;t=-0.631, P=0.538). Plasma sEPCR level in early onset group, late onset group and control group were (231 ± 11), (124±6)and(121±4)μg/L respectively, which is higher in early onset group than that in late onset group (t=48.080,P=0.001). There was no statistical difference of plasma sEPCR level between the late onset group and the control group(t=2.534,P=0.100). Conclusions The pathogenesis of early onset and late onset severe preeclampsia may be different. Decreased expression of EPCR in placenta may be associated with the pathogenesis of early onset severe preeclampsia.
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Objective: To explicit whether miR-200b can suppress the proliferation and invasion by targeting CD133, and to explore a molecular mechanism that miR-200b plays a role in tumor suppression in glioma. Methods: The CD133 3'-untranslated region (3'-UTR) mRNA-luciferase reporter vector was constructed and the dual-luciferase reporter gene assay was employed to examine the effect of miR-200b on activity of luciferase. U251 cells were transfected with miR-200b mimics and CD133-small interfering RNA (siRNA) (as a positive control) by LipofectAMINE 2000, and the expressions level of CD133 protein was detected by Western blotting. The inhibition effects of CD133 on cell proliferation and invasion were observed after CD133 siRNA were transfected into U251 cells. U251-SC cell mammosphere assay was performed after cotransfection with miR-200b mimics and CD133 siRNA. Results: miR-200b could bind to the 3'-UTR of CD133 and inhibit the activity of luciferase. CD133 protein expression was significantly down-regulated when miR-200b was overexpressed in U251 cells. Overexpression of miR-200b inhibited the invasion of U251 cells. Overexpression of miR-200b antagonized a role of proliferation and invasion in U251 cells. Overexpression of miR-200b reduced mammosphere number and the size of glioma stem cells. Conclusion: miR-200b can suppress cell proliferation and invasion by targeting CD133 in glioma. Copyright © 2014 by TUMOR.
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Objective To examine the effect of human cytomegalovirus (CMV) on the expressions of natural killer group 2 members (NKG2),including natural-killer group 2-member A (NKG2A),natural-killer group 2-member C (NKG2C) and natural-killer group 2 member D (NKG2D) receptors on the natural killer (NK) cells.Methods NK cells were isolated from the peripheral blood mononuclear cells of 20 healthy individuals using EasySep magnetic beads.NK cells and CMV-infected human embryonic lung fibroblast (HELF) were co-cultured for 24 h and regarded as experimental group.While CMV-uninfected NK cells co-cultured with HELF and only NK cells were taken as control groups.Flow cytometry was used to detect the expressions of the NK cell receptors in experimental and control groups.The comparison between groups was done by Wilcoxon rank sum test.Results The percentages of NK cells expressing NKG2C and NKG2D in group of CMV-infected NK cells co-cultured with HELF were 34.26％±7.99％ and 24.94％ ± 5.24％,respectively,which were significantly higher than those in CMVuninfected NK cells co cultured with HELF (26.31％ ±6.41％; Z=-3.285,P=0.001 and 20.02％±6.80％ ; Z=-3.285,P=0.001,respectively) and NK cells alone (25.78％± 6.62％ ; Z=-3.211,P=0.001 and 20.12 ％ ± 6.75 ％ ; Z=-3.285,P =0.001,respectively),while no significant changes were found in the expressions of the two receptors between groups of CMV-uninfected NK cells co-cultured with HELF and NK cells alone (both P＞0.05).Moreover,no significant changes were found in the expression of NKG2A among groups (all P＞0.05).The levels of NKG2A+ NKG2C,NKG2A+NKG2D,NKG2C+ NKG2A-and NKG2D+ NKG2A-cells in CMV infected NK cells co-cultured with HELFgroup were 2.99％ ±2.62％,6.13％±2.44％,26.62％ ±7.63％ and 20.44％±5.68％,respectively,while those in group of CMV-uninfected NK cells co-cultured with HELF were 4.44％ ± 4.25％ (Z=-2.240,P=0.025),6.99％±2.33％ (Z=-2.053,P=0.040),19.52％±6.80％ (Z=-2.800,P=0.005) and 15.78％±7.48％ (Z=-2.875,P=0.004),respectively.However,no significant changes were found in the expressions of NKG2A+ NKG2C+ and NKG2A+ NKG2D+ among groups (all P＞0.05).Conclusions CMV infection may change the NK receptor expressions,then stimulatory NKG2C and NKG2D signal transductions are enhanced and the inhibitory NKG2A signal conduction is weakened during CMV infection.
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Objective To evaluate the performance of sST 2 ELISA kit and investigate the clinical application of sST2.Methods This verification study validated the precision , linearity of sST2 ELISA kit according to the CLSI EP-15A, EP-6A protocols.300 healthy adults(aged from 20 to 85, 124 male and 176 female) from 5 different districts of Shanghai were used to establish serum sST 2 reference interval .The correlations between sST2, NT-ProBNP, LVEF and NYHA class were analyzed in 117 patients diagnosed with heart failure who were grouped according to the New York heart association ( NYHA).Receiver operating characteristic (ROC) curve was used to compare the ablity of sST2, NT-ProBNP, LVEF in distinguishing heart failure patients .Results The within-lot and between-lot variation of three level samples were below 4% and 10% respectively.There was a good linear correlation ( Y=0.995X+0.005, R2 =0.999) between theoretical value and actual detection result in the range of 0 to 200 μg/ml.The reference interval of sST2 was 10.2 to 41.0μg/ml for males and 8.9 to 28.1μg/ml for females.sST2 was positively correlated with NT-ProBNP and NYHA class but did not correlate with LVEF in heart failure patients . Patients with NYHA class>II (Median:28.3,IQR:19.5-39.2)had higher serum sST2 level than patients with NYHA class≤II (Median:45.1,IQR:34.1 -85.6), PII were 0.743, 0.810, 0.831 respectively and the sensitivity of sST 2 +NT-ProBNP+LVEF was 94.7%.Conclusions Experimental results show that this sST 2 ELISA kit has a good performance in the precision, linearity.sST2 correlates with NT-ProBNP and NYHA class but do not correlates with LVEF . Serum sST2 level is not influenced by age , BMI, renal function.sST2 could be a good supplement of NT-ProBNP and LVEF in distinguishing patients between NYHA class≤II and>II.
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Soluble ST2 (sST2) is a decoy receptor of IL-33 in blood.sST2and ST2L (receptor on cell membrane) seem to be markedly induced in mechanically overloaded cardiac myocytes,especially the expression of sST2.A large number of sST2 bind IL-33 thus subtracting IL-33 from the interaction with ST2L,potentially attenuating the cardioprotective effects of IL-33/ST2L pathway.Recently,sST2 has emerged as a novel for heart failure biomarker.Though sST2 was a less robust marker for the diagnosis of heart failure than NT-proBNP,it has been implicated in the prognostication of patients with acute dyspnea,acute or chronic heart failure and myocardial infarction,with particular value for mid-and long-term prognosis.However,further studies are needed in order to better point out the evidence for a routine use of sST2 evaluation in patients suffering from cardiovascular diseases.
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Objective To evaluate the role of phosphatidylinositol 3-kinase (PI3K) p110β in spinal dorsal horn neurons in the development of arthritic pain (AP) in rats and the relationship with transient receptor potential vanilloid 1 (TRPV1) and acid-sensing ion channel (ASIC)1 a.Methods Forty adult female Sprague-Dawley rats in which intrathecal catheters were successfully placed,aged 3 months,weighing 250-300 g,were randomly divided into 4 groups (n =10 each):control group (group C),group AP,AP + PI3K p110β missense oligo-deoxynucleotide group (group MS) and AP + PI3K p110β antisense oligo-deoxynucleotide group (group AS).AP was induced by injecting complete Freund's adjuvant into the ankle joint cavity of right hindpaw.Normal saline 20 μl,missense oligo-deoxynucleotide 15 μg (20μl) and antisense oligo-deoxynucleotide 15 μg (20 μl) were administered intrathecally once a day for 6 consecutive days starting from the time immediately after arthritis was induced in groups AP,MS and AS,respectively.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured 1 day before operation (T0) and on days 4,7,10 after operation (T1-3).The rats were then sacrificed after the last measurement of pain threshold at T3.L4-6 segment of the spinal cord was removed for detection of expression of PI3K p110β (by Western blot),and TRPV1 and ASICla (by immunohistochemistry)in spinal dorsal horn neurons.Results Compared with group C,MWT and TWL were significantly decreased atT1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was up-regulated in the other 3 groups(P＜ 0.01).MWT and TWL were significantly higher at T1-3,and the expression of PI3K p110β,TRPV1 and ASIC1a was lower in group AS than in groups AP and MS (P ＜ 0.01).Conclusion PI3K p110β in spinal dorsal horn neurons is involved in the development of AP in rats,and the mechanism is related to up-regulation of TRPV1 and ASIC1a expression in spinal dorsal horn neurons.
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Objective To investigate the effect of specific small interfering RNA (siRNA) targeting receptor of advanced glycation end products (RAGE) on the production of fibrosis markers (laminin,hyaluronic acid (HA) and N-terminal procollagen Ⅲ propeptide (PⅢ NP) in hepatic fibrosis (HF).Methods The expression vectors of specific siRNA targeting RAGE were constructed.Primary rat hepatic stellar cells (HSC) were isolated and cultured.The primary rat HSC were transfected with the recombinant vector.The blank control group and unspecific siRNA vector pAKD-NC-transfected group were as controls.The expressions of RAGE,laminin,HA and PⅢ NP at mRNA and protein levels were detected by real-time polymerase chain reaction and Western blot,respectively.Least-significant difference (LSD) and Student-Newman-Keuls (SNK) were performed to analyze standard normal distribution or homogeneous variance.Non-normal distribution and heterogeneity of variance data were analyzed by non-parametric Wilcoxon test.Results The expressions of RAGE at mRNA and protein levels in pAKD-GR126-transfected primary HSC were (42.32 ± 6.16)％,(43.24±7.50)％,(51.06±13.79)％ and (47.94±5.36)％ in blank control group and pAKDNC group (F=7.791 and 36.513,all P＜0.05).The expressions of laminin at mRNA and protein levels were (41.07±3.13)％,(40.59±5.87)％,(53.89±2.25)％ and (52.46±4.68)％ in blank control group andpAKD-NC group (F=225.111 and 88.039,all P＜0.05).The expressions of HA at mRNA and protein levels were (45.69 ± 0.87) ％,(46.08 ± 2.36) ％,(54.20 ± 0.56) ％ and (52.30±3.42)％ in blank control group and pAKD-NC group (F=178.317 and 180.646,all P＜ 0.05).The expressions of PⅢ NP mRNA at mRNA and protein levels were (56.10±4.18)％,(55.15±2.39)％,(54.40±2.79)％ and (53.58±6.18)％ in blank control group and pAKD-NC group (F=141.633 and 49.670,all P＜0.05).Conclusion RAGE specific siRNA could inhibit the expression of RAGE in primary rat HSC and could significantly lower the expression of fibrosis markers laminin,HA and PⅢ NP at mRNA and protein level.
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Objective To investigate the relationship between placental endoglin expression and the pathogenesis of severe preeclampsia (SPE). Methods Forty nine pregnant women with SPE and 40 normal pregnant controls were collected in Peking University First Hospital from January 2006 to January 2008,among which,nine SPE patients complicated with fetal growth restriction (FGR),six with hemolysis,elevated liver enzymes and low platelets syndrome (HELLP syndrome) and 12 with heavy proteinuria. The expression of placental endoglin was detected and semi-quantified by immunohistochemistry.Data were analyzed by independent sample t test. Results Endoglin was presented on cell membranes of placental syncytiotrophoblasts and vascular endothelial cells. The endoglin density of SPE group was higher than that of control group (0.1621± 0.0029 vs 0.1576 ± 0.0038,t=- 6.367,P＜0.05).No significant difference in endoglin density was found between FGR group and non-FGR group (0.1611±0.0026 vs 0.1623±0.0029,t=1.107,P＞0.05) ; neither did the heavy proteinuria group and non-heavy proteinuria group (0.1611±0.0032 vs 0.1624±0.0027,t=1.350,P＞0.05).The endoglin density of HELLP group was lower than that of non-HELLP group (0.1595±0.0032 vs 0.1625±0.0027,t=2.495,P=0.016). Conclusions The elevated placental endoglin expression might contribute to the pathogenesis of SPE.
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Objective To investigate the dynamic expression of Nogo-A receptor (NgR) in the hippocampus following focal cerebral ischemia in rats.Methods The focal cerebral ischemia model was established by electrocoagulating the right middle cerebral artery in rats.The expression of NgR in the ischemic hippocampal CA1,CA2 and CA3 regions at different time points after cerebral ischemia were detected by immunohistochemical staining.Results The expressions of NgR were up-regulated in hippocampal CA1 and CA2 regions at the first day,in the CA3 region at the second day after cerebral ischemia; the expressions of NgR in the CA1 and CA2 regions at the fifth day was decreased to the lowest.The expressions of NgR was up-regulated again in the CA1 and CA2 regions at the 28th day.Conclusions The NgR expression in the hippocampus in ischemic side showed different change characteristics at different regions,however,the overall change trend showed a 2 peaks and a valley phenomenon,which indicated that NgR might have different functions at different time periods after cerebral ischemia.
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ObjectiveTo investigate the effects of emulsified isoflurane preconditioning on the expression of platelet-activating factor (PAF) and PAF receptor during focal cerebral ischemia-reperfusion (I/R) in rats.MethodsThirty-two healthy adult male SD rats weighing 250-300 g were randomly divided into 4 groups ( n =8each):group sham operation (group S); group I/R; group emulsified isoflurane preconditioning( group EI) and group lipid emulsion (group LE).Focal cerebral I/R was induced by 2 h occlusion of middle cerebral artery followed by reperfusion in groups I/R,EI and LE.8％ emulsified isoflurane 10.5 ml/kg and 30％ lipid emulsion 10.5 ml/kg were injected intraperitoneally at 24 h before cerebral ischemia in groups EI and LE respectively.The neurologic deficit score (NDS) (0 =no deficit,4 =unable to control) was evaluated at 12 h of reperfusion.Venous blood samples were collected for measurement of plasma PAF concentration.The animals were then sacrificed and their brains removed for determination of infarct size (by TTC staining) and PAF receptor expression in hippocampus and cerebral cortex (by Western blot).ResultsFocal cerebral I/R significantly increased NDS,the infarct size,plasma PAF concentration and PAF receptor expression in cerebral cortex and hippocampus in group I/R as compared with group S.Emulsified isoflurane preconditioning significantly attenuated the focal cerebral I/R induced above changes in group EI as compared with group I/R,but there was no significant difference between groups I/R and LE.ConclusionEmulsified isoflurane preconditioning can attenuate focal cerebral I/R injury by inhibiting PAF and PAF receptor expression.
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Objective To investigate the expression of transforming growth factor-β1 (TGF-β1),endoglin (Eng) and their mRNA in placenta and superficial myometrium of patients with gestational hypertension or preeclampsia and to explore the role of Eng and TGF-β1 in the pathogenesis of gestational hypertension or preeclampsia.Methods One hundred and ten pregnant women were selected in the Second Affiliated Hospital of Tianjin Medical University from April 2009 to April 2010 who underwent cesarean sections and were divided into four groups:gestational hypertension group (n=30),mild preeclampsia group (n=30),severe preeclampsia group (n=30) and control group (normal pregnant women without labor and perinatal complications,n=20).The tissues of placenta and superficial myometrium were collected during cesarean section.Protein levels of Eng and TGF-β1 were detected by Western Blot.Real-time fluorescence reverse transcription polymerase chain reaction was used to detect the Eng and TGF-β1 mRNA expression.One-way ANOVA was used to compare among groups,Student-Newman-Keuls test was used to compare the differences between groups; relation between groups was analyzed by Pearson relation and linear regression.Results In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng mRNA level was 1.00,1.27±0.58,1.54±0.41 and 1.83±0.35,and the TGF-β1 mRNA level was 1.00,1.64 ± 0.33,1.92± 0.38 and 2.23 ± 0.53 in placenta respectively; those figures changed to 1.00,1.32±0.46,1.59±0.37 and 1.93±0.52,and 1.00,1.71 ± 0.45,1.91 ± 0.51 and 2.37 ± 0.46 in superficial myometrium respectively.The Eng and TGF-β1 mRNA levels of control group were lower than those of the other three groups (P＜0.05).The higher the mRNA level,the more severe the disease (P＜ 0.05).In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng protein expression was 0.11±0.07,0.15± 0.05,0.18 ± 0.06 and 0.43 ± 0.04,and the TGF-β1 protein expression was 0.11 ±0.02,0.26 ± 0.05,0.27± 0.03 and 0.88 ± 0.09 in placenta respectively; those figures changed to 0.14±0.06,0.16±0.04,0.20±0.08 and 0.46±0.05,and 0.15±0.03,0.29±0.06,0.31±0.04 and 0.91 ±0.08 in superficial myometrium respectively.The Eng and TGF-β1 protein levels of control group were lower than those of the other three groups (P＜0.05).The higher the protein level,the more severe the disease (P＜0.05).In the gestational hypertension group,mild and severe preeclampsia group,there were positive correlations between Eng and TGF-β1 protein levels in placenta (r=0.57,0.61 and 0.60 respectively,P＜0.05) and superficial myometrium (r=0.59,0.62 and 0.61 respectively,P ＜ 0.05).Conclusions Eng and TGF-β1 might play a role in pathogenesis of gestational hypertension and preeclampsia.
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Objective To study expression and activation of p38 mitogen activated protein kinase (MAPK) in vascular endothelial cells dysfunction in preeclampsia. Methods From Sept. 2009 to Mar.2010, 54 pregnant women underwent deliveries in the First Affiliated Hospital of Chongqing Medical University were enrolled in this study, including 20 patients in mild preeclampsia group, 16 patients in severe preeclampsia group and 18 women with term cesarean section without perinatal complications as control group. Placental endothelial cells were labeled by CD34 to assay microvessel density (MVD) of each group. Immunohistochemical SP and western blot were used to detect localization and expression of p-p38 MAPK protein, respectively. The levels of sera soluble fms-like tyrosine kinase-1 (sFlt-1)and soluble endoglin(sEng) were measured by ELISA. Results ①The MVD of placenta were 103 ± 3 in control group, 81 ±5 in mild preeclampsia group and 63±4 in severe group, respectively, which showed statistical difference among each group (P＜0.05).②The expression of p38 MAPK protein were 0.84±0.05 in control group,0.90±0.14 in mild group and 0. 86 ±0.18 in severe group, which did not reach remarkable difference among each group (P＞0.05). The expression of p-p38 MAPK protein were 0.13±0.05 in control group,0.59±0.12 in mild group and 1.16±0.18 in severe group, which show statistical difference among each group(P＜0.05).(3) The localization of p-p38 was in trophoblast, endothelial cells and a few (5.2±0.3)and(10.9±0.4)μg/L in control group,(12.5±1.2) and (20.4±5.3)μg/L in mild group and (19.3±3.0) and (29. 5 ±3.7) μg/L in severe group. When drawing paired comparison in those p-p38 MAPK protein levels and the concentrations of serum sFlt-1, sEng in preeclampsia groups (r=0.68,P＜0.05;r=0.87,P＜0.05). Conclusions The remarkable activation of the p38 MAPK in the placenta of patients with preeclampsia induced the increased levels of sFlt-1 and sEng in maternal serum, which confer the injury of vascular endothelial cells that caused the significant decline of MVD in placentas. p38 MAPK signaling might be one of the key pathways in vascular endothelial cell dysfunction in preeclampsia.
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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.
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Toll like receptors(TLRs) , a pattern conservative receptor superfamily are found on mammalian cell surface over the last decade. TLRs recognize distinct pathogen - associated molecular patterns, which can lead not only to the responses of the innate immunity but also to the development of the antigenspecific adaptive immunity. The expression of TLRs have changed in some carcinomas, besides, microbial composition and drugs mediate the tumor immunotherapy by the way of TLRs signal transduction. Consequently , deep studies on TLRs will provide new approaches for tumor immunotherapy.
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Objective To explore the ralationship between maternal serum level of soluble endoglin (sEng) in advanced gestations and hypertensive disorders comlicating pregnaney(HDCP). Methods The serum levels of sEng were analyzed using enzyme-linked immunosorbent assay (ELISA). Blood samples were obtained from 62 pregnant women with HDCP at 35-39 weeks' gestation (20 gestational hypertension, 20 mild preeclampsia, 19 severe preeclampsia and 3 eclampsia), and 20 normal pregnant women at 37-39 weeks' gestation (control). Results The serum sEng levels in normal, gestational hypertension, mild preeclampsia, severe preeclampsia and eclampsia group were (6.24±0. 26) ng/ml; (6. 56±0. 29) ng/ml; (7.47±0. 31) ng/ml; (8. 71± 0. 37) ng/ml and (9.69±0. 28) ng/ml, respectively. The serum sEng levels in the preeclampsia and eclampsia group were significantly higher than those in the gestational hypertension and normal group (P<0. 01), that of the severe preeclampsia group was significantly higher than the mild preeclampsia (P<0. 01), and that of the eclampsia group was significantly higher than the preeclampsia group (P<0. 01). However, no difference was found between the gestational hypertension and normal group (P>0. 05). Conclusions The increased serum level of sEng may participate in the genesis of HDCP.