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 construct in the extracellular matrix metalloproteinase inducer (EMMPRIN)glycosylation single point mutation plasmid,and to explore its relationship with tumor cell proliferation.Methods PCR point mutantion technology was used to construct the mutantion plasimid of EMMPRIN glycosylation single point.After successful mutation, the function of mutantion plasmids were detected. Western blotting was used to detect the expression of EMMPRIN protein,immunofluorescence method was used to detemine the morphological changes of the cells,and MTT assay was performed to detect the relationship between mutantion pasmid and tumor cell proliferation.Results Confirmed by restriction enzyme digestion and sequencing,the 44th,the 152th,and the 186th Asn were successfully mutated to Gln in the sequence of EMMPRIN;EMMPRIN/GFP (N44Q), EMMPRIN/GFP(N152Q),and EMMPRIN/ GFP (N186Q)glycosylation single point mutation plasmids were constructed.Compared with wild-type,thel morphology of the cells was significantly changed,the core division of mutant-type cells was significantly reduced, the number of filopodia was reduced. The results of MTT assay showed that the survival rate of the cells in wild-type group were significantly increased compared with control group (P<0.05 );the survival rates of the cells in EMMPRIN (N44Q) group, EMMPRIN (N152Q) group and EMMPRIN(N186Q)were significantly decreased compared with wild-type group(P<0.05).Conclusion Mutant-type EMMPRIN can inhibit the proliferation of tumor cells;with the duration increasing, the inhibitory effect is weakened.There is a correlation between EMMPRIN glycosylation and proliferation of tumor cells.

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.

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

Objective To observe the expression of hypoxia inducible faetor-1α (HIF-1α) in the retina of rabbits with acute high intraocular pressure and to investigate the mechanism of systemic domestic recombinant human erythropoietin (rhEPO) protecting the retina from ischemia-reperfusion injury. Methods First, control group and model group were established in rabbit eyes. The acute high intraocular pressure model was established by saline perfusion into anterior chamber, and then hypodermic injection of domestic rhEPO was made. HIF-1α protein in the retina was observed by immunohistochemical staining method on days 1, 3, 7 and 14 after retinal ischemla-reperfusion, respectively. Results No cells with HIF-la positive expression were observed in the retina of the control group. Ceils with HIF-1α positive expression in the model group outnumbered those in the control group (P < 0. 01). The resemblance pattern occurred in EPO group but its degree was slightly greater than that in the model group from day 3 after ischemia-reperfusion (P<0.05). Conclusion Domestic rhEPO can down-regulate the expression of HIF-1α in the retina with acute high intraocular pressure, which may be one of the mechanisms that rhEPO protects the retina from ischemia-reperfusion injury.

Objective To explore the stress status and its influencing factors of nursing interns in an military hospital.Methods 190 nursing interns were surveyed by an self-made general information questionnaire and a pressure source scale of practice nurses.Results The stress status at the end of practice of nursing interns in our hospital was at the low level.The total pressure source score was (25.37 ± 4.69).The major influencing factors of stress status were levels of education (technical secondary school,college or bachelor degree) and manners of profession choosing ( voluntarily,family intention or assigned).Among them,interns with higher level of education had more stress.Interns who chose nursing as profession by assignment had higher stress level than those by family intention or voluntarily.Conclusions Understanding basic characteristics and special situations of nursing interns with different education levels and manners of profession choosing is useful for nursing managers to make effective intervention strategies,adjust and protect stress level,and ensure physical and mental health of the nursing interns.

OBJECTIVETo investigate the effect of noninvasive positive pressure ventilation( NIPPy) on the gene and protein expression of biquitin-proteasome of skeletal muscle in patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD).

METHODSSeven patients with AECOPD by NIPPV were used as the study group, meanwhile, 6 patients with AECOPD who refused NIPPV was the control group. The blood gas analysis, heart rate (HR) and mean arterial pressure (MBp) were monitored before and 14 days after treatment. A skeletal muscle biopsy was performed after 14 days of therapy. The mRNA expression of ribosomal protein S21 (RPS21), Ubiquitin, Ubiquitin combined with enzyme E2 (E2), Ubiquitin ligase E3 (E3) in skeletal muscle cell were measured by RT-PCR. The protein expression of mitochondrial aconitase (AC02), protease C3 (C3), ribosomal protein SLC16 (SLC16) were detected by Western blot.

RESULTSForteen days after treatment, the patients in NIPPV group got much better improvement in PaCO2, PaO2 and HR than that of the patients.in the control group (P < 0.05). The gene expression of RPS21,Ubiquitin, E2 and E3 in skeletal muscle cell on patients with NIPPV were obviously lower than that of the control group (P < 0.05, P < 0.01). Compared with that of the control group, the protein expression of C3 and AC2 increased significantly in the NIPPV group (P < 0.01). The protein expression of SLC16 was significantly lowered in the treated group (P < 0.01).

CONCLUSIONNIPPV can ameliorate the proteasome pathway and energy metabolic disorders in patients with AECOPD.

Humans ; Muscle, Skeletal ; metabolism ; Positive-Pressure Respiration ; Proteasome Endopeptidase Complex ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; therapy ; Ubiquitin ; metabolism

Objective To establish high-performance liquid chromatography with electrochemical detector(HPLC-ECD) method for the determination for metanephrine and normetanephrine in 24 h urine, and provide a superior test for the diagnostic of pheochromocytomas over plasma/urine catecholamine.Methods MCX solid-phase cartridge was used for extraction of metanephrine and normetanephrine,HPLC-ECD was used for their measurements.The intra-assay CVs,interassay CVs and recoveries of metanephrine and normetanephrine were also calculated.104 hypertensive patients without pheochromocytomas and 5 pheochromocytomas patients were selected in this study.The concentrations of metanephrine and normetanephrine were compared with the plasma and 24h urinary catecholamines concentrations.Results The intra-assay CV,inter-assay CV and recovery of metanephrine were 5.9%, 7.5%,91.1% respectively;the intra-assay CV,inter-assay CV and recovery of normetanephrine were 6.3%,6.6%,88.5%,respectively.The MN,NMN,plasma CA and urine CA of all pheochromocytomas patients were positive.MN and NMN were negative in controls,while plasma CA and urine CA are false positive in 15 patients and 14 patients in controls,respectively.Conclusions The study establish a fast and accurate method for quantification of metanephrine and normetanephrine in 24 h urine by HPLC-ECD.These findings also prove that it is the best biochemical assays for pheochromocytomas at present.