Objective To discuss the relationship between plasma levels of tissue factor(TF) in the circulation and the development of smooth muscle cells(SMC) hyperplasia after percutaneous coronary intervention(PCI) in porcine models.Methods Thirty swines were used and their coronary arteries were injured by balloon dilatation.Morphological measurements included neointimal area and the extent of area stenosis.Plasma TF level was measured before and after PCI.The relationship between TF and neointimal area was calculated.Results Different degrees of intimal hyperplasia were found among all vessels and the rate of restenosis was 60%.Plasma TF levels were significant higher in vessels which subsequently developed restenosis(30 minutes after PCI: 688.36?91.57 ng/L vs 514.87?68.73 ng/L,24 hours after PCI: 706.09?89.59 ng/L vs 539.22?70.51 ng/L,P

Objective To construct pcDNA3.1 RASSF1 eukaryotic vector and observe the influence of RASSF1 on the apoptosis of hepatocarcinoma cell line HepG2. Methods RASSF1 gene was amplifled from human RASSF1 cDNA by polymerase chain reaction (PCR) and cloned into pcDNA3.1. The recombinant plasmid pcDNA3. 1 RASSF1 was transfected into hepatocarcinoma HepG2 cell line. The expression of RASSF1 was examined by Western blot. The influence of RASSF1 on the cell apoptosis was measured by Annexin V/PI assay. Results DNA enzyme digestion and sequencing results showed that recombinant plasmid pcDNA3. 1-RASSF1 was successfully constructed. RASSF1 protein was overexpressed in HepG2 cell line transfected with pcDNA3. 1-RASSF1 plasmid. The apoptosis rate of blank, pcDNA3. 1 and pcDNA3. 1-RASSF1 group was (5.8 ±0.42)%, (7.48 ±0.68)% and (35. 1 ±3. 15)%, respectively.Conclusion The pcDNA3. 1- RASSF1 eukaryotic vector was successfully constructed, RASSF1 protein overexpression could induce apoptosis in HepG2 cell line.

Objective To improve the diagnosis and treatment of acute exacerbation in idiopathic pulmonary fibrosis(IPF)Methods A case with clinical proven acute exacerbation in IPF and treated with corticosteroid was described and the foreign literature was reviewed.Results Most patients with idiopathic pulmonary fibrosis(IPF)show slowly progressive deterioration.However,accelerated deterioration also occurs without any definite factors on infection,heart failure,et al in a few of patients with IPF who have previously shown slowly progressive deterioration,and for part of patients corticosteroid treatment is effective.Conclusion Some acute exacerbation in idiopatic pulmonary fibrosis can be idiopathic.

Sonic hedgehog (Shh) signaling has recently been shown to be involved in the pathological angiogenesis in response to tissue hypoxia and ischemic injury. Hypoxia/ischemia is considered to play an important role in the development of choroidal neovascularization (CNV). This study was aimed to examine the effect of blockade of the Shh signaling pathway on CNV and the underlying mechanism. A total of 64 male Brown-Norway (BN) rats were used in this study. One eye of each rat underwent laser photocoagulation. The other eye served as normal control. After the laser treatment, the 64 rats were divided into four groups (n=16 in each group): Blank control group, in which no intravitreal administration was given; cyclopamine group, recombinant Shh N-terminals protein (rShh) group and phosphate-buffered saline (PBS) group, in which cyclopamine (a Shh inhibitor), rShh (a Shh activator) and PBS were intravitreally injected into the laser-treated eyes respectively every other day for a total of four intravitreal injections immediately after the laser treatment. Fourteen days after the intravitreal administration, the changes of CNV-related variables, including positive CNV lesion percentage, CNV membrane area and CNV membrane thickness, were evaluated by fluorescein angiography, indocyanine green angiography and pathological examinations. The mRNA and protein expression of PTCH1, Gli1, HIF-1(α), VEGF and DLL4 in each group on 14 days of CNV model was detected by real-time quantitative PCR and western blot analysis, and the relationship between the Shh cascade and the HIF-1(α)-VEGF-DLL4 cascade in CNV was analyzed. The results showed that the CNV membrane area and the CNV membrane thickness were decreased by 62.5% and 41.9% in the cyclopamine group and increased by 85.7% and 64.3% in the rShh group in comparison to those in the blank control group (P<0.01 for each). There was no significant difference in the CNV membrane area and thickness between the blank control group and PBS group (P=0.102 and P=0.063, respectively). Real-time quantitative PCR revealed a 5.23-, 4.14-, 2.97-, 2.78- and 2.39-fold up-regulation of the mRNA expression of PTCH1, Gli1, HIF-1(α), VEGF and DLL4 genes in the laser-treated eyes compared with the normal control eyes in the control group. In the cyclopamine group, the mRNA and protein expression of Gli1, HIF-1(α), VEGF and DLL4 was significantly down-regulated (P<0.05 for each) while the expression of PTCH1 showed no significant changes at the mRNA (P=0.293) and protein level (P=0.304). The mRNA expression and protein expression (P=0.001 and P=0.021, respectively) of PTCH1, Gli1, HIF-1(α), VEGF and DLL4 was significantly increased in the rShh group when compared with the control group. The expression level of these genes was related to the severity of the CNV. It was concluded that intravitreal administration of cyclopamine can effectively inhibit the formation of laser-induced experimental CNV by down-regulating the expression of the HIF-1(α)-VEGF-DLL4 cascade in CNV. The Shh signaling pathway as an upstream signaling pathway of HIF-1(α)-VEGF-DLL4 cascade is implicated in the development of experimental CNV.

Objective To discuss the methods and outcomes of twin reversed arterial perfusion sequence in different stage. Methods From August 2012 to December 2013, 11 cases were diagnosed with twin reversed arterial perfusion sequence (TRAP) by 3d color Doppler ultrasound in the Maternal ang Fetal Medicine Center ,Affiliated Shengjing Hospital, China Medical University, including 3 cases in stageⅠa, 7 in stageⅡa and 1 case in stageⅡb. We performed expected therapy to 3 cases in stageⅠa and 4 cases in stageⅡa [1 case (case 9) in stageⅡa developed to stageⅡb and was terminated by emergency cesarean section], radio frequency ablation (RFA) selective reduction to 3 cases in stage Ⅱa, emergency cesarean section to 1 case in stageⅡb. All cases except case 1,4 took cesarean section to terminate pregnancy and would be followed up on the aspect of infant′s growth. Results (1) 3 cases in stageⅠa were monitored by sequential ultrasound examination, all donors grew normally, in which 2 acardias arised spontaneous cessation of flow and were implemented of cesarean section after the normal period of gestation; 1 case suffered premature rupture of membrane and experienced the natural childbirth at 34 gestation weeks plus 5 days. The survived average labor weight was 2 923 g. (2) In the 7 stage Ⅱa cases, 3 cases were implemented of RFA selective reduction, 2 cases survived and the average labor gestation was 35 weeks plus 1 day and average labor weight was 2 050 g;in 3 expected therapy cases, 1 case suffered premature rupture of membrane at 34 gestation weeks plus 5 days and experienced cesarean section; 1 case was observed for 3 weeks and the donor was stillborn; 1 case progressed to stage Ⅱb and experienced cesarean section at 34 gestation weeks plus 5 days;and 1 case was lost of follow up. (3) For 2 cases of stageⅡb cases, one was progressed from stage Ⅱa during expected therapy; the other one had been already in stage Ⅱb since visiting and experimented an emergency cesarean section, but the neonate died of heart failure two days after labor. (4) 7 survived neonates were followed up, all had a normal condition compared to kids of the same age, except one suffering a congenital heart disease, patent ductus arteriosus and atrial septal defect and whose weight (6 kg at 7 months) was lighter. Conclusions We should make the diagnosis of TRAP as early as soon and choose appropriate therapy according to the stage. According to our research, cases in stage Ⅰa can undergo an expected therapy under a sequential ultrasound examination;cases in stageⅡa can undergo a RFA selective reduction to improve the prognosis of the donor, which can make a good perinatal outcome.

Intravascular stent as a foreign body exhibits obvious reaction following implantation,such as activation of coagulation system and elevation of serum C-reactive protein,inflammatory marker.After platelet is activated,P-selectin binds to the glycoprotein ligand 1 expressed on leukocyte,and forms congeries on the leukocyte.In addition,interleukin-1?,interleukin-6 and tumor necrosis factor-? are greatly secreted.The two factors are main reasons for restenosis following stent implantation.Studies on spiral CT angiography and transcranial Doppler sonography are important.

Objective To analyze the clinical outcome and impact factors of twin-to-twin transfusion syndrome (TTTS) with anterior placenta treated by fetoscopic selective laser coagulation of placental vessels(SLCPV). Methods Ten cases of TTTS with anterior placenta and 8 cases with posterior placenta were treated by SLCPV in Shengjing Hospital from July 2011 to April 2014. Clinical data were analyzed retrospectively. Some cases were at Quintero stageⅡor higher stage, others were at Quintero stageⅠbut with cardiovascular score≥5 according to the scoring system of Children′s Hospital of Philadelphia. The anterior placenta cases were treated by curve fetoscopy and the posterior placenta cases were treated by straight fetoscopy. Results (1) Of all the 18 cases, the mean gestational age at SLCPV was 24.1 weeks (17+5 to 27+4 weeks). There were 2 cases at Quintero stageⅠ, 4 at stageⅡand 12 at stageⅢ. The mean gestational age of anterior placenta cases at SLCPV was 25.2 weeks (22 to 27+4 weeks), with 2 cases at Quintero stageⅠ, 2 cases at stage Ⅱ and 6 at stage Ⅲ;5 cases had preterm prelabour rupture of the membranes (PPROM)and 1 case had maternal intestinal obstruction after the operation;the average operation time was 40 minutes. Of the posterior placenta cases, the mean gestational age at SLCPV was 22.7 weeks (17+5 to 27+4 weeks);2 cases were at Quintero stageⅡand 6 cases at stageⅢ. PPROM happened in one case;one case had maternal enterobacter cloacae septicemia;the average operation time was 28 minutes. All the 18 cases could tolerate the operations. There was no intraoperative complication. (2) One anterior placenta case had maternal intestinal obstruction and miscarriage; and one posterior placenta case had enterobacter cloacae septicemia. Karyotype analyses of the all the twins were normal. (3) 17 cases delivered already, including all the 10 anterior placenta cases and 7 posterior placenta cases. One infant had corpus callosum agenesis, but its co-twin was normal. The average gestational age at delivery for anterior placenta cases was 32.6 weeks (24 to 37+1 weeks), an was 28.2 weeks (25+6 to 36+2 weeks) for posterior placenta cases. The fetuses survival rate was 13/17 (one case was still in pregnancy) for at least one twin, and 10/17 for both twins. Both twins survival rates were 5/10, 5/7 for anterior placenta cases and posterior placenta cases, respectively. At least one twin survival rates were 8/10, 5/7 for the two groups respectively. Conclusion SLCPV is suitable for the treatment of TTTS, no matter the placenta is on the anterior wall or posterior wall. The treatment had good outcomes, but more PPROM happened in the anterior placenta cases, which may be associated with operation time and the range of operation.

Objective To observe the effect of RNA interference binding immunoglobulin protein (BiP) on expression of bone markers and keytranscription factors in osteoblast exposed to fluoride.Methods MC3T3-E1 cells were used as osteoblast model in vitro.The cell viability was test with cell counting Kit-8 after cells were administrated with varying concentrations of fluoride [0.0 (control),0.1,1.0,2.0,4.0,8.0,16.0,20.0,32.0 and 64.0 mg/L] for different duration.Cells transfected with small interfering RNA (siRNA) BiP were exposed to fluoride (2.0,8.0 and 20.0 mg/L) for 2 days.Real-time PCR and Western blotting technique were used to determine the gene and protein levels of BiP.Meantime,the mRNA expression of bone markers and key transcription factors was investigated by real-time PCR.Results The difference of all viability in fluoride-dose groups was statistically significant exposed for 1,3,7 and 14 days (F =46.7,118.6,214.6,325.6,all P ＜ 0.05).Expression of BiP significantly increased in cells exposed to 20.0 mg/L compared to that of control (11.22 ± 3.25 vs.7.94 ± 1.31,P ＜ 0.05).The expression of alkaline phosphatase (ALP) elevated in cells exposed to 2.0 mg/L of fluoride (12.81 ± 3.62 vs.6.86 ± 2.13,P ＜ 0.01);conversely,it significantly reduced in cells exposed to 20.0 mg/L of fluoride (0.89 ± 0.17 vs.6.87 ± 2.14,P ＜ 0.01).Cells transfected with siRNA BiP significantly decreased the ALP expression in cells exposed to fluoride compared to that of cells only exposed to the same concentration of fluorine (12.81 ± 3.62 vs.7.43 ± 2.06;5.92 ± 2.38 vs.3.96 ± 0.21,all P ＜ 0.05).Cells transfected with siRNA BiP and administrated with 2.0 mg/L significantly reduced the osteocalcin expression (4.29 ± 0.99 vs.1.29 ± 0.86,P ＜ 0.01).Similarly,expression of runt-related transcription factor 2 (Runx2) significantly increased in cells exposed to 2.0 mg/L.However,expression of Runx2 significantly decreased in cells transfected with siRNA BiP and administrated with fluoride.Cells transfected with siRNA BiP significantly decreased the Runx2 expression in cells exposed to 2.0 mg/L and 20.0 mg/L compared to that of cells only exposed to the same concentration of fluorine (1.13 ± 0.22 vs.6.61 ± 0.48;0.02 ± 0.02 vs.1.50 ± 0.38,all P ＜ 0.01).As the downstream of Runx2,the expression of osterix in cells treated with different concentrations of fluoride was similar to that of Runx2.Conclusion BiP is not directly involved in the process of osteoblast differentiation induced by fluoride;instead,it affects the expression of bone markers and key transcription factors in osteoblast exposed to fluoride.

Objective To explore the neutrophil to lymphocyte ratio (NLR) and its relationship with the effect of chemotherapy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL).Methods The clinicopathological characteristics and outcome of 51 patients with DLBCL diagnosed by pathological biopsy and immunohistochemistry who received CHOP or R-CHOP regimen were collected and reviewed.According to the median of NLR,the patients were divided into low NLR group (NLR≤2.32) and high NLR group (NLR＞2.32).The prognostic influence of the NLR on overall survival (OS) was studied by Kaplan-Meier method and Log-rank test.To evaluate the independent prognostic relevance of NLR,univariate and multivariate Cox regression models were applied.Results The complete response (CR) rates of the low and high NLR groups were 71.4 ％ (20/28) and 39.1％ (9/23),respectively (P =0.02).The OS in the low NLR group was significantly better than that in the high NLR group (1,2 and 3-year OSs were 96.4 ％,90.4 ％ and 72.3 ％ vs 63.9 ％,52.7 ％ and 42.2 ％,respectively,P =0.009).Univariate and multivariate Cox regression models analysis showed that NLR ＞ 2.32 was an independent prognostic factor (P =0.016).Conclusion An elevated NLR before treatment indicates the poor effect of chemotherapy and prognosis of patients.NLR is an independent prognostic factor for DLBCL.

Objective To discuss the characteristics of clinical features,diagnosis and therapy procedure of Anti-gamma-aminobutyric acid B (anti-GABAB)receptor encephalitis.Methods Procedure of clinical diagnosis and treatment in two patients with anti-GABAB receptor encephalitis were retrospected in Neurological Department of Tianjin Huanhu Hospital and the characteristics of anti-GABAB receptor encephalitis were analyzed with literature review.Results In both patients,the clinical manifestations were epileptic seizure,which was a focal seizure at first,seconded by generalized tonic clonic seizure,with frequent attacks or epilepticus state.Significant cognitive and memory impairments and psychological and behavioral abnormalities were observed during the course of the disease.Abnormal MRI signals were found in mesial temporal lobe,with normal electroencephalography.Antibody against anti-GABABreceptor was positive in both cerebrospinal fluid and blood,and immune regulation therapy was effective.Conclusions The patients present with epileptic seizures,seconded by cognitive and memory impairment,often accompanied by MR imagingrevealed mesial temporal lobe lesion.And positive anti-GABAB receptor antibody is major characteristics of anti-GABAB receptor encephalitis.The patients often combine small cell lung cancer,and Immune regulation therapy is effective.