Background and purpose:Loss or altered expression of Ras association domain family 1A gene (RASSF1A) through DNA methylation has been associated with the pathogenesis of a variety of cancers, which suggests the tumor suppressor function of this gene. This study aimed to explore the effect of DNA methyltransferase inhibitor 5-Aza-2’deoxycytidine (5-Aza-dc) on demethylation and expression of RASSF1A in cervical cancer cell lines. Methods:HPV positive cervical cancer cell lines HeLa and Caski, HPV negative cell lines HT-3 and C-33A were treated with two different concentration of 5-Aza-dc (5 μmol/L, 10 μmol/L). MSP (methylation-specific PCR) and Bisulfite genomic sequencing PCR (BGS) combined with TA clone were used to investigate methylation status of RASSF1A gene promoter and exon 1 before and after treatment of 5-Aza-dc. RASSF1A gene mRNA expression was detected by RT-PCR. Results:Two HPV positive cell lines showed hypomethylated RASSF1A promoters and expressed RASSF1A mRNA, and after treatment with 5-Aza-dc, the mRNA expression of RASSF1A did not change significantly (FHeLa=3.003, P=0.125; FCaski=0.045, P=0.956). Two HPV negative cervical cancer cell lines showed hypermethylation status of RASSF1A promoter and silenced RASSF1A. After treatment with 5-Aza-dc, demethylation occurred in the promoter region of RASSF1A gene, which subsequently induced re-expression of this gene in HT-3 and C-33A. The F test (FHT-3=18.002, P=0.03;FC-33A=17.179, P=0.03) and LSD-t test (P<0.05) demonstrated that significant difference in the expression of RASSF1A was found upon two different concentrations drug treatment.Conclusion:The methylation status of promoter and exon 1 of RASSFIA gene in HPV positive and HPV negative cervical cancer cell lines are different. The promoter hypermethylation is correlated with RASSF1A gene expression in HPV negative cervical cancer cell line HT-3 and C-33A, and plays a key role in RASSF1A silencing. 5-Aza-dc may effectively reverse the methylation status of RASSFIA gene promoter in cervical cancer HT-3 and C-33A cells and reactivate gene expression silenced by aberrant hypermethylation in a dose-dependent manner within certain extent.

Objective To study the effect of sub-chronic exposure to deltamethrin(DM) on the behavior of mice in learning and memory and expression of GABAA receptor α1 and γ2 subunits in the hippocampus of mice.Methods 60 female SPF Kunming mice were randomly divided into solvent control group,low-dose group,middle-dose group and high-dose group, 15 mice in each group.The latter three groups were exposured to deltamethrin for 60 days by gavage.Open field test (OFT) was applied to evaluate locomotor activity and exploratory behavior in mice.RQ-PCR was employed to measure the expression of GABAA receptor α1 and γ2 subunits in hippocampus of mice.Results After exposure to DM,the moving distance of the central area in the middle-dose group ((555.1 ± 12.8) cm) and high-dose group ((516.4± 11.88) cm) was significantly higher than that in the solvent control group ((327.3± 117.8) cm, P＜0.05).Numbers of standing in marginal area (F=4.117, P=0.010) and total movement distance (F=2.914, P=0.042) in the high-dose group ((27.9±9.9) times, (3211.3±379.8) cm) were also significantly higher than that in the solvent control group ((15.1 ±8.9)times, (3211.3±379.8)cm).The expression of GABAA receptor α1 subunit mRNA in the middle-dose group and high-dose group was significantly lower than that in the low-dose group and solvent control group(F=8.508, P=0.001) and the expression of GABAA receptor γ2 subunit mRNA in high-dose group was significantly lower than that in the other groups (F=6.738, P=0.002).Conclusion Sub-chronic exposure to DM can damage the function of locomotor activity and exploratory behavior,and inhibit the expression of GABAA receptor α1 and γ2 subunits in the hippocampus of mice.

Objective To study the effect of sub-chronic exposure to dehamethrin(DM) on the behavior of mice in learning and memory.Methods 60 Female SPF Kunming mice were randomly divided into 4 groups and given DM by gavaging for 60 days.Morris water maze (MWM) was used to evaluate spatial memory in mice.Results After exposure to DM,the escape latency of the solvent control group and the treatment groups were (12.20±6.5)s,(14.99±5.4) s,(15.64±8.3)s,(22.71±6.2)s on the fifth day.The escape latency of the high-dose group was higher than those of the low-dose group (P=0.0041) and the solvent control group (P=0.019) in the navigation test.The number of crossing position of the platform in the high-dose group ((2.93± 1.53)times) and the middle-dose group ((3.40± 1.12) times) were lower than that in the solvent control group ((5.87 ± 1.55) times) and the low-dose group ((4.90± 1.41)times)(P＜0.05).Conclusion Sub-chronic exposure to DM can damage the spatial learning and memory of mice.

Objective To observe the management and outcome of the cardiac tamponade patients during the ablation procedure using two different anticoagulation strategies. Methods All the patients developed tamponade during the ablation procedure were enrolled from January 2007 to December 2013 in our center. In group 1, warfarin was discontinued 3 to 5 days before the procedure and low molecular weight heparin (LMWH) was administered subcutaneously until ablation procedure day. In group 2, warfarin was not discontinued and the international normalized ratio INR was to maintained between 2 and 3. Results There were 27 patients (0.6%) developed cardiac tamponade out of a total 4487 patients received ablation in our center. The baseline clinical characteristics including age, left atrium, the heparin dose and ACT during the procedure had no signiifcant difference between the groups, except that the INR was higher in the group 2 (0.9±0.1 vs. 2.3±0.5, P<0.001). There was no signiifcant difference in the amount of pericardiac drainage between the two groups (365±222 ml vs. 506±300 ml, P=0.137). Two patients in group 1 patient (11.1%) and 1 in group 2 (11.1%) needed emergency surgical repair (P>0.999). The median hospital day was similar in the 2 groups [(9.6±3.3) d vs. (12.1±4.5) d, P=0.167]. There were no other serious complications and no hospital death. Conclusions Non-discontinuation of warfarin during peri-procedural catheter ablation of AF is not signiifcantly different to bridging with LMWH in the management and outcome of acute cardiac tamponade.

Objective Identify the mechanism of induced atrial arrhythmias after pulmonary vein isolation (PVI) in patients with paroxysmal atrial ifbrillation(PAF), and investigate its long-term prognosis. Methods All patients with PAF undergoing PVI and induction test afterwards between Feburary 2010 and October 2010 were included. The induction protocol was rapid pacing initiated at cycle length of 250 ms with progressive shortening in a decrement of 10 ms down to 180 ms or refractoriness. Isoproterenol of 2-4μg/min was administrated as well. Inducibility was deifned as induction of atrial arrhythmia lasting >1 min. The mechanism of induced tachycardia was identiifed by activation mapping and entrainment mapping under the guidance of CARTO system. All patients were followed up by 36 months. Results Forty-nine atrial tachycardia were induced in 39 (19.7%) patients, including 35 organized atrial tachycardia (OAT) and 14 atrial ifbrillation (AF). The LA diameter was signiifcantly larger in inducible group than non-inducible group (39.5±6.6 mm vs. 36.7±5.2 mm, P=0.004). Macroreentry was the most common mechanism in induced OATs (28, 80.0%), and mitral isthmus was the most common critical site (20, 40.8%), followed by cavo-tricuspid isthmus (12, 24.5%), PV (6, 12.2%), LA septum (4, 8.2%), superior vena cava (3, 6.1%) and LA roof (1, 2.0%). Conclusions The most common mechanism of induced tachycardia by IV isoproterenol and rapid pacing is MI and CTI dependent after PVI in PAF patients, which can be succssefully eliminated by liner ablation, not increasing long-term recurrence rate.

Objective To study the risk factors for postoperative nausea and vomiting (PONV) in gynecological patients administered prophylactic antiemetics.Methods A total of 1 000 gynecological surgical cases were reviewed.Data on patient characteristics,anesthesia,surgeries,and 1st postoperative day nausea and vomiting were collected and analyzed statistically.Results Multivariate logistic regression analysis included a history of motion sickness,the use of an analgesic pump,the duration of anesthesia,and the use of neostigmine.The odds ratios (ORs),in decreasing order,were that of a history of motion sickness [OR =1.953,95％ confidence interval (CI):1.404-2.716],the use of neostigmine (OR =1.831,95％ CI:1.203-2.786),the use of an analgesic pump (OR =1.472,95％ CI:1.017-2.129),and the duration of anesthesia (OR =1.256,95％ CI:1.058-1.492).The predictive model was described by the following equation:P=1/(1+e(1623-0.669X1-0.386X2-0.228X3+0.605X4)).Tne predictive accuracy of this model for the incidence of PONV in gynecological patients was 70.6％,and the area under the receiver operating characteristic curve for the predictive result was 0.642.Conclusion The independent risk factors for PONV in gynecological patients administered prophylactic antiemetics are a history of motion sickness,the use of neostigmine,the use of an analgesic pump,and the duration of anesthesia,in decreasing order of significance.

Objective To evaluate and compare the effectiveness of propofol-remifentanil closed-loop and open-loop anesthesia in gynecological laparoscopic operation under bispectral index (BIS) monitoring.Methods Forty female patients undergoing elective gynecological laparoscopic operation were recruited and randomly divided into closed-loop (group Ⅰ) and open-loop (group Ⅱ) groups.During anesthesia maintenance,the closed-loop group was administered with a BIS-feedback system to regulate the target effect-site concentration;whereas,the open-loop group was administered the target effect-site concentration according BIS value manually.The variation of non-invasive mean arterial pressure (MAP),heart rate (HR),SpO2,BIS,extubation time,consumption of propofol and remifentanil,Ramsay index,and subjective comfort grade were recorded at the selected time points.Results The extubation time in group Ⅰ was shorter than in group Ⅱ.The total dosage of propofol administered in group Ⅰ was less than that in group Ⅱ,but there was no significant difference in the total consumption of remifentanil.HR in group Ⅰ was steadier than in group Ⅱ.However,the MAP and Ramsay index were similar in both the groups.The subjective comfort grade in group Ⅰ was higher than in group Ⅱ.Conclusion The use of propofol-remifentanil closed-loop system by BIS-feedback anesthesia is safer,more controllable,with higher degree of satisfaction and sparing side-effects,we therefore recommend it during gynecological laparoscopic operations.

Objective Cryoballoon ablation of pulmonary vein (PV) ostia often induces a vagal response.This prospective study was designed to assess the effectiveness of prophylactic intravenous administration of atropine on hemodynamic impairment induced by cryoballoon ablation in patients with atrial fibrillation.Methods Twenty-five patients with paroxysmal atrial fibrillation undergoing cryoballoon ablation were prospectively enrolled and assigned to either the trial group on the control group.First twelve patients (the trial group) were administered 1 mg of atropine before deflation of the cryoballoon,while the following 13 patients (the control group) were given atropine only after the onset of the hemodynamic variation (decrease in heart rate and/or blood pressure).Treatment was considered effective when the hemodynamic variations were restored.Results In the trial group,three patients with transient hypotension did not require further supportive care throughout the procedures and one patient with hypotension required supportive management.In the control group,hypotension,bradycardia and mixed bradycardia with hypotension requiring supportive care occurred in six,three,and three patients,respectively.Overall,the rate of marked vagal responses was significantly lower when prophylactic atropine was administrated (4/12 vs.12/13 patients,P ＜ 0.01).Conclusions Atropine is effective in the prevention of all types of vasovagal responses induced by cryoballoon ablation in patients with atrial fibrillation.

Objective:To evaluate the value of autologous platelet-rich plasma (aPRP) separation-retransfusion for blood conservation in the patients undergoing thoracolumbar laminectomy.Methods:Sixty American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-60 yr, with body mass index of 19-30 kg/m 2, scheduled for elective thoracolumbar laminectomy, were divided into 2 groups ( n=30 each) using a random number table method: conventional blood conservation group (group C) and aPRP blood conservation group (group aPRP). Group C received tranexamic acid and autologous blood salvage-retransfusion. Group aPRP received aPRP separation-retransfusion, tranexamic acid and autologous blood salvage-retransfusion. The volume of allogeneic blood transfused, percentage of patients who did not need the allogeneic blood transfusion and adverse reactions were recorded. Venous blood samples were collected for blood routine examination and for determination of the plasma concentrations of interleukin-6 (IL-6) and IL-10 at 1 day before operation (T 0), when the volume of blood loss reached 500 ml (T 1), immediately after surgery (T 2), and at 24 and 48 h after surgery (T 3, 4). The incidence of hypoxemia and amount of 24-h wound drainage were recorded. Results:Compared with group C, the amount of allogeneic red blood cells, plasma transfused and 24-h wound drainage were significantly decreased ( P<0.05), the percentage of patients who did not need the allogeneic red blood cell and plasma transfusion was increased (30% vs 47%, 10% vs 60%, P<0.05), the plasma concentrations of IL-6 and IL-10 at T 2-4 were significantly decreased ( P<0.05), and the incidence of hypoxemia in PACU was decreased in group aPRP (27% vs 10%, P<0.05). Conclusions:aPRP separation-retransfusion can provide marked improvement in conventional blood conservation in the patients undergoing thoracolumbar laminectomy.

Background::Oral anti-coagulants (OAC) are the intervention for the prevention of stroke, which consistently improve clinical outcomes and survival among patients with atrial fibrillation (AF). The main purpose of this study is to identify problems in OAC utilization among hospitalized patients with AF in China.Methods::Using data from the Improving Care for Cardiovascular Disease in China-Atrial Fibrillation (CCC-AF) registry, guideline-recommended OAC use in eligible patients was assessed.Results::A total of 52,530 patients with non-valvular AF were enrolled from February 2015 to December 2019, of whom 38,203 were at a high risk of stroke, 9717 were at a moderate risk, and 4610 were at a low risk. On admission, only 20.0% (6075/30,420) of patients with a diagnosed AF and a high risk of stroke were taking OAC. The use of pre-hospital OAC on admission was associated with a lower risk of new-onset ischemic stroke/transient ischemic attack among the diagnosed AF population (adjusted odds ratio: 0.54, 95% confidence interval: 0.43–0.68; P <0.001). At discharge, the prescription rate of OAC was 45.2% (16,757/37,087) in eligible patients with high stroke risk and 60.7% (2778/4578) in eligible patients with low stroke risk. OAC utilization in patients with high stroke risk on admission or at discharge both increased largely over time (all P <0.001). Multivariate analysis showed that OAC utilization at discharge was positively associated with in-hospital rhythm control strategies, including catheter ablation (adjusted odds ratio [OR] 11.63, 95% confidence interval [CI] 10.04–13.47; P <0.001), electronic cardioversion (adjusted OR 2.41, 95% CI 1.65–3.51; P <0.001), and anti-arrhythmic drug use (adjusted OR 1.45, 95% CI 1.38–1.53; P <0.001). Conclusions::In hospitals participated in the CCC-AF project, >70% of AF patients were at a high risk of stroke. Although poor performance on guideline-recommended OAC use was found in this study, over time the CCC-AF project has made progress in stroke prevention in the Chinese AF population.Registration::ClinicalTrials.gov, NCT02309398.