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Objective@#To investigate the changes and clinical significance of vascular endothelial (VE)-cadherin and procalcitonin (PCT) in serum and cerebrospinal fluid (CSF) of children with viral encephalitis or bacterial meningitis(BM).@*Methods@#A total of 42 cases of children with viral encephalitis(viral encephalitis group), 36 cases of children with BM(BM group), and 20 cases of children with non-nervous system injury(control group) were selected from September 2016 to June 2018 at the Third Hospital of Zhengzhou University.The serum and CSF levels of VE-cadherin and PCT levels of the 3 groups were detected by using enzyme-linked immunosorbent assay.@*Results@#The levels of VE-cadherin in the serum of viral encephalitis group, BM group and control group at the acute phase were (5.60±1.17) mg/L, (7.08±1.01) mg/L and (2.52±0.68) mg/L respectively, and the levels of VE-cadherin in CSF of viral encephalitis group, BM group and control group were (6.00±1.09) mg/L, (6.97±1.11) mg/L and(1.93±0.88) mg/L, respectively.The levels of PCT in the serum of viral encephalitis group, BM group and control group at the acute phase were (0.26±0.11) μg/L, (0.82±0.17) μg/L and (0.27±0.13) μg/L, respectively, and the levels of PCT in the CSF of viral encephalitis group, BM group and control group were (0.25±0.11) μg/L, (0.72±0.14) μg/L, (0.28±0.17) μg/L, respectively.As a result, the levels of VE-cadherin and PCT in the serum and CSF of BM group showed significant increase, compared with viral encephalitis group and control group in the acute phase(F=124.94, 163.21, 151.62, 127.37, all P<0.01). The levels of VE-cadherin in the serum and CSF of viral encephalitis group were also significantly higher than that of control group (all P<0.01), but there was no difference between viral encephalitis group and control group about the levels of PCT in the serum and CSF (all P>0.05). The levels of VE-cadherin in the serum of viral encephalitis group and BM group after treatment were (2.34±0.81) mg/L and (2.67±1.29) mg/L, and were(2.55±0.92) mg/L and(2.39±0.74) mg/L in the CSF.The levels of PCT in the serum of viral encephalitis group and BM group after treatment were (0.25±0.11) μg/L, (0.30±0.17) μg/L, and the levels of PCT in the CSF of viral encephalitis group and BM group were(0.22±0.10) μg/L and (0.27±0.12) μg/L.After effective treatment, the levels of VE-cadherin, PCT in serum and CSF of BM group and viral encephalitis group were almost equal, and the difference was not statistically significant(F=1.83, 0.76, 2.72, 3.89, all P>0.05). In the receiver operating characteristic (ROC) curve, the area under the ROC curve of VE-cadherin in serum and CSF was 0.896 and 0.912, and was 0.670 and 0.668 of PCT respectively.@*Conclusions@#VE-cadherin may be involved in the early stage of intracranial infection, and it may be helpful in differentiation of virus or bacterial infection with PCT.VE-cadherin has a good diagnostic value for intracranial infection.
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Objective To investigate the changes and clinical significance of Caveolin-1,matrix metalloproteinase-9 (MMP-9) and interleukin-1β (IL-1β) in cerebrospinal fluid of children with bacterial meningitis or viral encephalitis.Methods Thirty-six cases of children with bacterial meningitis,42 cases of children with viral encephalitis,and 20 cases of children with non-nervous system infection were selected from September 2016 to June 2018 at the Third Affiliated Hospital of Zhengzhou University.The levels of Caveolin-1,MMP-9 and IL-1β in cerebrospinal fluid were detected by using enzyme linked immunosorbent assay (ELISA).Results Cerebrospinal fluid Caveolin-1,MMP-9,IL-1β levels in the acute phase of bacterial meningitis were(49.06 ± 8.96) ng/L,(134.79 18.88)μg/L,(100.02 ± 14.67) μg/L,respectively,and (29.13 ± 7.25) ng/L,(18.69 ± 7.23) μg/L,(47.57 ± 8.95)pg/L in recovery phase,which were higher than those of the controls [(11.18 ± 2.24) ng/L,(11.53 ± 3.54) μg/L,(39.75 ± 7.08) μg/L)],and the differences were significant (all P ＜ 0.05).Cerebrospinal fluid Caveolin-1,MMP-9,IL-1β levels in the acute phase of viral encephalitis were (42.71 ± 10.48) ng/L,(62.78 ± 17.39) μg/L,(57.97 ± 11.28) μg/L,respectively,and (29.13 ± 7.25) ng/L,(18.69 ± 7.23) μg/L,(47.57 ± 8.95) μg/L in recovery phase,which were higher than those of controls,and the differences were significant (all P ＜ 0.05).The levels of Caveolin-1,MMP-9 and IL-1β in cerebrospinal fluid of bacterial meningitis group and viral encephalitis group were significantly higher than those of convalescent group (all P ＜ 0.05).The levels of Caveolin-1,MMP-9,IL-1β in cerebrospinal fluid of bacterial meningitis group were significantly higher than those in viral encephalitis group (all P ＜ 0.05) in the acute phase,and no significant difference was found in the recovery phase(all P ＞ 0.05).Cerebrospinal fluid Caveolin-1,MMP-9,IL-1β showed no significant difference among children with different severity of intracranial infection.Correlation analysis showed that there was a positive correlation between Caveolin-1,MMP-9 and IL-1 β levels in cerebrospinal fluid of acute in bacterial meningitis group and viral encephalitis group (Caveolin-1 and MMP-9:R2 =0.239,P ＜ 0.05;MMP-9 and IL-1β:R2 =0.766,P ＜0.01;Caveolin-1 and IL-1β:R2 =0.245,P ＜ 0.05).Conclusions Caveolin-1,MMP-9 and IL-1 β involved in the pathogenesis of intracranial infection in children,and the effects of different pathogens on intracranial infection were different.
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Objective@#To investigate the changes and clinical significance of Caveolin-1, matrix metalloproteinase-9(MMP-9) and interleukin-1β(IL-1β)in cerebrospinal fluid of children with bacterial meningitis or viral encephalitis.@*Methods@#Thirty-six cases of children with bacterial meningitis, 42 cases of children with viral encephalitis, and 20 cases of children with non-nervous system infection were selected from September 2016 to June 2018 at the Third Affiliated Hospital of Zhengzhou University.The levels of Caveolin-1, MMP-9 and IL-1β in cerebrospinal fluid were detected by using enzyme linked immunosorbent assay(ELISA).@*Results@#Cerebrospinal fluid Caveolin-1, MMP-9 , IL-1β levels in the acute phase of bacterial meningitis were(49.06±8.96) ng/L, (134.79±18.88) μg/L, (100.02±14.67) μg/L, respectively, and (29.13±7.25) ng/L, (18.69±7.23) μg/L, (47.57±8.95) μg/L in recovery phase, which were higher than those of the controls[(11.18±2.24) ng/L, (11.53±3.54) μg/L, (39.75±7.08) μg/L)], and the differences were significant (all P<0.05). Cerebrospinal fluid Caveolin-1, MMP-9, IL-1β levels in the acute phase of viral encephalitis were (42.71±10.48) ng/L, (62.78±17.39) μg/L, (57.97±11.28) μg/L, respectively, and (29.13±7.25) ng/L, (18.69±7.23) μg/L, (47.57±8.95) μg/L in recovery phase, which were higher than those of controls, and the differences were significant (all P<0.05). The levels of Caveolin-1, MMP-9 and IL-1β in cerebrospinal fluid of bacterial meningitis group and viral encephalitis group were significantly higher than those of convalescent group (all P<0.05). The levels of Caveolin-1, MMP-9, IL-1β in cerebrospinal fluid of bacterial meningitis group were significantly higher than those in viral encephalitis group (all P<0.05) in the acute phase, and no significant difference was found in the recovery phase(all P>0.05). Cerebrospinal fluid Caveolin-1, MMP-9, IL-1β showed no significant difference among children with different severity of intracranial infection.Correlation analysis showed that there was a positive correlation between Caveolin-1, MMP-9 and IL-1 β levels in cerebrospinal fluid of acute in bacterial meningitis group and viral encephalitis group(Caveolin-1 and MMP-9: R2=0.239, P<0.05; MMP-9 and IL-1β: R2=0.766, P<0.01; Caveolin-1 and IL-1β: R2=0.245, P<0.05).@*Conclusions@#Caveolin-1, MMP-9 and IL-1 β involved in the pathogenesis of intracranial infection in children, and the effects of different pathogens on intracranial infection were different.
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Objective:To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1(MDR1)and the in vitro expansion of CD34+ cells derived from umbilical cord blood.Methods:CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and co-cultured with AFT024 cells(AFT024 group)or cultured alone(control group)for 7 days.During the subsequent 14 days,retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells.On the 7th,14th and 21st day after culture,the number of total nucleated cells(TNC)was counted,the ratio of CD34+ cells was assayed by flow cytometry(FCM)and the number of CD34+ cells was calculated,and colony-forming cells(CFC)were counted by methylcellulose cultures.RT-PCR method was used to detect the level of MDR1 mRNA in the transfected cells.The expression and function of P-glycoprotein(P-gp)were evaluated by FCM assay and Rhodamine-123 efflux assay,respectively.The gene transfection efficiency was calculated by drug-resistant colony-forming cells assay.Results:(1)The MDR1 mRNA level in AFT024 group than that in control group.The gene transfection efficiency in AFT024 group was significantly higher than that in control group(46.0% vs 15.2%,P0.05).On the 14th day,the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P0.05).The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P
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AIM: To explore the feasibility of using vector-based small interfereing RNA(siRNA) to inhibit the expression of MDR1 mRNA and P-glycoprotein and to reverse the multidrug resistance of drug-resistant ovarian cancer cell line.METHODS: An adriamycin-resistant human ovarian cancer cell subline OVCAR/AR was established by stepwise inducement.Another mutidrug-resistant human ovarian cancer cell subline OVCAR/MDR was established by transfecting multidrug resistant gene 1(MDR1) into ovarian carcinoma cell line OVCAR-3.Transfection of MDR1 mRNA specific siRNA expressing plasmids(pSN/mdr1a and pSN/mdr1b) into OVCAR/AR and OVCAR/MDR cells was performed using liposome transfection reagents.MDR1 mRNA expression level was quantified using real time reverse transcription polymerase chain reaction(real time RT-PCR).Flow cytometry(FCM) was performed to assess the expression of p-glycoprotein(P-gp).Multidrug resistant to anticancer agents was evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay.RESULTS: Basal MDR1 mRNA expression level in drug-resistant cell line OVCAR/MDR was higher than that in OVCAR/AR cell line,and was both higher than that in its parent cell line OVCAR-3.The expression of MDR1 mRNA and P-gp protein in both OVCAR/AR and OVCAR/MDR cells was inhibited dramatically by transfecting with pSN/mdr1a and pSN/mdr1b.The reversal rate of chemoresistance to adriamycin in OVCAR/AR and OVCAR/MDR transfected with pSN/mdr1a and pSN/mdr1b was 79.5% and 93.9%,compared with control group.CONCLUSION: MDR1 expression in the drug-resistant cell lines is partially inhibited by treatment with vector-based MDR1 specific small interference RNAs at the mRNA and protein level,which increases the chemotherapy sensitivity of these drug resistant ovarian carcinoma cell sublines.