Objective:To investigate the role of miRNA-4298/PADI4/p53 signal axis in mediating 4f-induced apoptosis of leukemia cells.Methods:The cell growth density was observed under inverted microscope and the proliferation of leukemia cells was detected by CCK-8 counting assay. The expression of PADI4 and P53 at mRNA level was detected by qRT-PCR. Cell cycle and apoptosis were measured with flow cytometry. The expression of PADI4, P53, Bcl-2, Bax, caspase-3 and caspase-9 at protein level was detected by Western blot. Differential miRNA and mRNA expression profiles was detected by next generation sequencing. Databases such as TargetScan were used to predict the potential upstream and downstream genes of PADI4. A luciferase reporter assay was used to detect the 3′UTR of PADI4 targeted by miRNA-4298. Cell transfection assay was used to detect the effect siRNA, PADI4 vector, miRNA mimics and miRNA inhibitor in interference and rescue.Results:Nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor 4f could inhibit the proliferation of THP-1, K562 and U937 cells, and induce the apoptosis of these leukemia cells. It downregulated the expression of PADI4 mainly through the binding activity of miRNA-4298 to miRNA sponges, which resulted in the proliferation inhibition and apoptosis of leukemia cells. The inhibited proliferation and apoptosis of leukemia cells by 4f were associated with the increase of P53 expression after the decrease of PADI4 expression. The PADI4-dependent upregulation of P53 led to the ratio inversion of downstream Bcl-2/Bax, which activated caspase-3 or caspase-9 to induce the apoptosis of leukemia cells.Conclusions:The apoptosis of leukemia cells induced by Nrf2 inhibitor 4f was mainly associated with the miRNA-4298/PADI4/p53 axis, suggesting that it might be a novel signaling pathway for targeted therapy.

Mutations of the X-linked methyl-CpG-binding protein 2 (MECP2) gene in humans are responsible for most cases of Rett syndrome (RTT), an X-linked progressive neurological disorder. While genome-wide screens in clinical trials have revealed several putative RTT-associated mutations in MECP2, their causal relevance regarding the functional regulation of MeCP2 at the etiologic sites at the protein level requires more evidence. In this study, we demonstrated that MeCP2 was dynamically modified by O-linked-β-N-acetylglucosamine (O-GlcNAc) at threonine 203 (T203), an etiologic site in RTT patients. Disruption of the O-GlcNAcylation of MeCP2 specifically at T203 impaired dendrite development and spine maturation in cultured hippocampal neurons, and disrupted neuronal migration, dendritic spine morphogenesis, and caused dysfunction of synaptic transmission in the developing and juvenile mouse cerebral cortex. Mechanistically, genetic disruption of O-GlcNAcylation at T203 on MeCP2 decreased the neuronal activity-induced induction of Bdnf transcription. Our study highlights the critical role of MeCP2 T203 O-GlcNAcylation in neural development and synaptic transmission potentially via brain-derived neurotrophic factor.
Animals ; Humans ; Methyl-CpG-Binding Protein 2/metabolism* ; Mice ; Neurodevelopmental Disorders/genetics* ; Rett Syndrome/genetics* ; Synaptic Transmission ; Threonine

Objective:To investigate the effect of self-made phase-change nanodroplets IR780/FA-Nds-DTX as molecular targeted ultrasound contrast agents for accurate diagnosis and combined targeted therapy of pancreatic cancer.Methods:Pancreatic cancer cell lines were cultured in vitro and 50 tumor bearing nude mouse models were established. The experimental group (IR780/ FA-NdS-DTX) and four control groups[ normal saline, Nds(FITC), FA-Nds (FITC) and IR-780] were divided to verify dual-mode targeted imaging. The imaging using the IVIS Imaging System verified the high-efficiency targeted detection ability and near-infrared fluorescence imaging of IR780/FA-Nds-DTX for tumors in vivo, phase transformation induced by low-intensity focused ultrasound and further contrast-enhanced ultrasound imaging verified the high-efficiency aggregation of IR780/FA-Nds-DTX in local tumors and accurate evaluation of tumor contour. The therapeutic effect was observed in the experimental group (IR780/FA-Nds-DTX) and four control groups (FA-Nds-IR780, FA-Nds-DTX, FA-Nds and normal saline). After low-intensity focused ultrasound irradiation for 30s induced microbubble blasting after phase transformation in each group, 808nm photothermal therapy apparatus was used to irradiate tumor area in each group. Two-dimensional ultrasound was used to monitor the changes in tumor volume in each group before and at 3 d, 9 d, 15 d after treatment, and the changes in tumor volume rate and inhibition in each group were statistically analyzed and compared.Results:The amount of IR780/ FA-Nds-DTX locally targeted aggregation was the largest, and the average fluorescence intensity of tumor in the experimental group was significantly higher than that of the control groups: IR780/ FA-Nds-DTX group compared with Nds(FITC) group[(5.12±0.69)×10 7 vs (1.06±0.23)×10 7, P<0.05], IR780/FA-Nds-DTX group compared with FA-Nds (FITC) group [(5.12±0.69)×10 7 vs (2.98±0.34)×10 7, P<0.05], IR780/FA-Nds-DTX group compared with IR-780 group [(5.12±0.69)×10 7 vs (1.54±0.42)×10 7, P<0.05], and there was no fluorescence in tumor area in saline group. Further contrast-enhanced ultrasound imaging after nanodroplet phase transformation could more accurately locate the tumor boundary. After 15 days of photothermal ablation combined with chemotherapy, the growth rate of tumor volume in the IR780/ FA-Nds-DTX treatment group was significantly lower than that in the control groups: IR780/FA-Nds-DTX group compared with FA-Nds-IR780 group[(0.105±0.075) vs (0.405±0.175), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds-DTX group [(0.105±0.075) vs (1.385±0.035), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds group [(0.105±0.075) vs (2.255±0.105), P<0.05], IR780/ FA-Nds-DTX group compared with normal saline group [(0.105±0.075) vs (2.185±0.155), P<0.05]. And the tumor inhibition rate increased significantly: IR780/ FA-Nds-DTX group compared with FA-Nds-IR780 group [(0.93±0.06) vs (0.48±0.17), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds-DTX group [(0.93±0.06) vs (-0.51±0.105), P<0.05], IR780/ FA-Nds-DTX group compared with FA-Nds group [(0.93±0.06) vs (-1.63±0.115), P<0.05], IR780/ FA-Nds-DTX group compared with normal saline[(0.93±0.06) vs (-1.35±0.245), P<0.05]. Conclusions:The self-made phase-change ultrasound contrast agents IR780/FA-Nds-DTX have good potential clinical value in targeted detection and combined therapy of pancreatic cancer with small lesions or even metastases.

Objective To restrospectively analyze the risk factors for blood transfusion during ce-sarean section and neonate outcomes. Methods A total of 291 parturients ( 60 cases with blood transfusion during surgery and 231 cases without blood transfusion during surgery) who underwent cesarean section from November 2016 to March 2017 in our hospital were selected. The significant covariates in one-way analysis of variance were analyzed by logistic regression analysis. The odds ratio ( OR) and 95％ confidence interval ( CI) were calculated. The risk factors for blood transfusion during cesarean section were identified, and neonate outcomes were analyzed. Results Placenta increta, placenta accrete and pernicious placenta pre-via were the risk factors for blood transfusion during cesarean section, with OR value ( 95％ CI ) 13. 5 (2. 6-56. 8), 6. 1 (2. 1-11. 6) and 3. 3 (1. 6-8. 6), respectively. The prolonged gestational age was a protective factor, and the OR value ( 95％ CI) was 0. 3 ( 0. 2-0. 5) . The duration of operation, anesthesia time and length of hospital stay were significantly prolonged in blood transfusion group as compared with non-blood transfusion group ( P<0. 05) . The body weight and 1-min Apgar score of newborns were signifi-cantly lower in blood transfusion group than in non-blood transfusion group. Conclusion Placenta increta, placenta accrete and pernicious placenta previa are the risk factors for blood transfusion during cesarean sec-tion. Blood transfusion is not helpful for neonate outcomes.

Objective To analyze the dose dependent cardiotoxicity of cyclophosphamide and thus establish the mouse models for understanding the underlying mechanisms . Methods Thirty mice ( 7 -8 weeks old) were randomly divided into 6 groups:control of one course group ,control of two courses group , one course of low dose ( 50 mg/kg) group ,two courses of low dose group ,one course of high dose ( 100 mg/kg) group ,two courses of high dose group . The changes of cardiac function and morphology were analyzed at indicated times by ultrasound and histology respectively . Results Compared with the control groups , injection of low dose cyclophosphamide either with one course or two courses ,and injection of high dose with only one course had no in statically significant changes in hair color ,physical activity ,body weight and cardiac function( P > 0 .05) . Treatment of two courses with high dose resulted in significant decrease of physical activity and body weight ( P < 0 .05 ) . Meanwhile ,cardiac systolic function was significantly decreased( P <0 .05) . Histology study revealed robust cardiac hypertrophy and fibrosis . Conclusions Two courses of intraperitoneal injection of cyclophosphamide at high dose result in obvious cardiotoxicity , especially systolic dysfunction .Intraperitoneal injection of two courses of cyclophosphamide serves as a good strategy to induce cardiotoxicity in mice ,which could be applied for future mechanism study .

Objective To evaluate the effect of propofol pretreatment on nuclear factor kappa B (NF-κB)activity during hepatic ischemia-reperfusion(I∕R)injury in rats. Methods Twenty-four patho-gen-free healthy male Sprague-Dawley rats, aged 2 months, weighing 200-250 g, were divided into 3 groups(n=8 each)using a random number table:sham operation group(group S),group I∕R and propofol pretreatment group(group P).Hepatic I∕R injury was induced by 45 min occlusion of the hepatic artery and portal vein entering the middle and left lobes of the liver followed by reperfusion. In group P, propofol 12 mg·kg-1·h-1was infused via the femoral vein until the end of ischemia starting from 30 min before ische-mia in group P. Blood samples were collected from the inferior vena cava at 120 min of reperfusion for deter-mination of the levels of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β). The rats were then sacrificed and livers were re-moved for determination of phosphorylated NF-κB p65(p-NF-κB p65)expression in liver tissues(by West-ern blot)and apoptosis in hepatocytes(by TUNEL).Apoptosis index(AI)was calculated. Results Com-pared with group S, the levels of ALT, AST, TNF-α and IL-1β in serum and AI were significantly in-creased,and the expression of p-NF-κB p65 in liver tissues was up-regulated in I∕R and P groups(P<005). Compared with group I∕R,the levels of ALT,AST,TNF-α and IL-1β in serum and AI were signifi-cantly decreased,and the expression of p-NF-κB p65 in liver tissues was down-regulated in group P(P<005). Conclusion The mechanism by which propofol pretreatment reduces hepatic I∕R injury is associat-ed with inhibiting NF-κB activity in rats.

Objective To propose an accurate method of noninvasive determination of central venous pressure(CVP ) by locating the central point of right atrium (RA ) using echocardiography .Methods Through the 3D reconstruction ,the accurate positions of RA of 30 patients who had been examined by multislice 3‐dimensional computed tomography for chest imaging were recorded .Based on solid geometric principles ,the central point in RA was located by echocardiography and then compared with CT‐location point .The accuracy and feasibility were assessed by absolute distance (Da) ,vertical distance (Dv) and the whole time of location (T) between the two points .Results Mean Da ,Dv and T of the whole subjects were 07.6cm(95% CI:06.2to08.1cm),01.6cm(95% CI:-00.2to03.4cm),and438.0s(95% CI:400.1to 47 4.0 s) ,respectively .Conclusions The echocardiographic method on the basis of solid geometry proposed in this study could be used to locate the central point in RA accurately and simply .Thus it would be helpful to improve the accuracy of noninvasive determination of central venous pressure .

Objective To determine the central venous pressure (CVP) noninvasively based on hemodynamics principles using ultrasound location of the collapse point of the internal jugular vein.Methods Forty patients were enrolled in this study.The collapse point of the internal jugular vein was located and marked by a linear transducer,the body mark of right atrium was marked on the right lateral wall of the chest.The noninvasive CVP was calculated according to the vertical distance between those two points.The invasive CVP determination by central venous catheter was also carried out on all the patients.Correlation analysis was used to compare the invasive and noninvasive methods.With invasive determination of CVP as the gold standard,the ROC curve of the noninvasive ultrasound method was sketched to explore the optimal cut-off points.Results The correlation analysis reveal high positive correlation between CVPs determined by ultrasound imaging and central venous catheter (r =0.906,P ＜0.01).By the ROC curve test,fluid column height of 10.75 cm by ultrasound method was determined as the cut-off point,with the sensitivity and specificity of diagnosing elevation of CVP being 88.9％ and 93.5 ％ respectively.The corresponding area under the curve was 0.971.Conclusions Ultrasound imaging could be used to determine CVP noninvasively,which would be helpful in diagnosis of the circulating load of patients.

Objective To analyze the levels and clinical significances of IL-18,Caspase-3 and nerve tissue-specific protein S-100B at different disease extent and different stages of infants with hypoxicischemic encephalopathy (HIE).Methods This study was clinical experimental studies.Sixty-seven infants with HIE (23 cases of mild HIE,23 cases of moderate HIE,21 cases of severe HIE) from February 2008 to June 2009 in Hebei Children's Hospital were enrolled.The levels of IL-18,Caspase-3 and S-100B protein in all samples were measured at acute phase (1 d,3 d) and recovery phase (7 d) by ELISA method.Twenty healthy full-term neonates were selected as the normal control group.Multi-factor analysis of variance and Pearson correlation test was used for statistical analysis.Results The levels of the three indicators in the moderate and severe group were higher than the normal control group.In the moderate group,IL-18 levels were(132.15 ± 9.87),(150.31 ± 15.04) and (87.91 ± 9.93) ng/L,Caspase-3 levels were (5.79 ±0.64),(7.36 ± 1.57)and (3.79 ±0.61) μg/L,S-100B levels were(6.82 ±0.61),(9.62 ± 1.29) and (10.76 ± 1.64) μg/L.In the severe group,IL-18 levels were (160.23 ± 16.03),(189.86 ± 18.32) and (107.35 ± 13.02) ng/L;Caspase-3 levels were (6.86 ± 1.02),(9.54 ± 1.43) and (5.25 ± 0.71) μg/L;S-100B levels were(8.90 ± 0.32),(12.54 ± 0.89)and(13.53 ± 0.75) μg/L.In the normal control group,IL-18 levels were (71.08 ± 11.52),(72.53 ± 11.05) and (71.93 ± 11.30) ng/L; Caspase-3 levels were (2.84 ± 0.52),(2.98 ± 0.53) and (2.87 ± 0.52) μg/L; S-100B levels were (1.50 ± 0.25),(1.62 ±0.30)and(1.53 ±0.29) μg/L IL-18 levels,Caspase-3 levels and S-100B levels in severe group were higher than the moderate group and the mild group were higher than the mild group.IL-18 levels were (73.46 ± 4.77),(77.59 ± 4.02) and (72.87 ± 6.92) ng/L ; Caspase-3 levels were (3.13 ± 0.31),(3.63±0.40) and (3.26 ±0.45) μg/L;S-100B levels were(3.68 ±0.40),(5.851 ±0.63) and(6.95 ± 0.58) μg/L in the mild group.S-100B levels in the mild group were higher than that in the normal control group.The IL-18 and Caspase-3 levels were risen in the third day to the first day in the acute phase of the moderate group and severe group,decreased in the recovery phase.Serum S-100B protein levels in the acute and recovery phase increased gradually,and there was no correlation between the three indicators (r-=0.321,0.14,0.48,P=0.438,0.974,0.911 respectively).Conclusions IL-18,Caspase-3 and S-100Bwere involved in the pathophysiological process of HIE.The levels were closely related to the severity and disease progression of HIE,the severer of the illness,and the higher of the levels.Dynamic monitoring the changes of the three indicators may contribute to an early diagnosis,condition monitoring and prognosis of HIE.

OBJECTIVE: To filtrate and prove the different microRNAs (miRs) profiles in nasopharyngeal carcinoma. METHOD: Screening the different expressions of miRs between nasopharyngeal carcinoma and the inflammatory tissues by the application of expression profiling of chip high-throughput and large-scale microarray analysis. Then we used RT-QPCR technology to prove the accuracy of screening results. RESULT: There were significant expression differences of miRs between nasopharyngeal carcinoma and the control tissues, 144 human miRs had 2 or more fold the difference ratio. Compared with the inflammatory tissues, we have found that miRs-34b, miRs-449b and miRs-7-1 significantly low expressed in nasopharyngeal carcinoma, yet miRs-125b, miRs-184, miRs-196b, miRs-205 and miRs-24-1 expressed high. The results were consistent with the microarray analysis. CONCLUSION The difference expressed miRs might be closely related to the process of nasopharyngeal carcinoma, and the research on miRs profiles maybe provide a powerful target basis for early diagnosis and therapy of nasopharyngeal carcinoma.
Carcinoma ; Gene Expression Profiling ; Humans ; MicroRNAs ; genetics ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; Oligonucleotide Array Sequence Analysis