Objective To construct the recombinant plasmid that highly expressed human subcellular PTEN,in order to provide a basis for further study on its anti-tumor effect. Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product was ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant expression vector DUM-PTEN and PUM-NSL PTEN were identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

Objective To investigate the effect of different forbidden drink schemes on anesthesia induction and postoperative nausea and vomiting during pediatric interventional heart surgery.Methods One hundred and twenty pediatric patients underwent cardiac interventional procedures under general anesthesia were randomized into group A,B and C,with 40 patients in each group.Patients in cach group were fasted for 8 h preoperatively.Water was prohibited for patients in group A for 6 h preoperatively.Patients in groups B and C orally took 10％ glucose solution and a multivitamin drink (Outfast) at 2 h preoperatively,respectively.Nausea and vomiting after anesthesia induction were recorded for 24 h postoperatively.Results The sedation and mask acceptance scores were significantly higher in group B and C than those in group A,and were significantly higher in group C than those in group B (P ＜ 0.05,resoectively).Following anesthesia induction,MAP was significantly higher in group B and C than that in group A (P ＜ 0.05,resoectively).Both the severity and incidence of postoperative nausea and vomiting were higher in group B and C than those in group A,and was lower in group C than that in group B (P ＜ 0.05,resoectively).Conclusions Taking clear water orally 2 hours before surgery can decrease the restlessness of pediatric interventional heart surgery during anesthesia induction period,with stable smooth hemodynamics,reducing the degree of PONV.

Objective:To explore the clinical characteristics, mechanism and countermeasures of drug-induced second thrombocy-tosis ( ST) in order to ensure safe medication. Methods:Drug-induced ST published during 1996 and 2016 in domestic and overseas medical journals was collected and statistically analyzed in respects of age, gender, kind and distribution of drug-induced ST, treatment and outcome of ST. Results:Totally 198 cases were collected, which involved 30 different drugs of eight types mainly including antitu-mor drugs, blood system drugs and antibiotics. Among the patients, 112 ones (56. 6％) were male and the other 86 ones (43. 4％) were female. Totally 14 patients had embolization with the overall incidence of 7. 0％. Conclusion:Without timely and favorable treat-ment, drug-induced ST may lead to thrombosis. To ensure the safety of medication, platelet level should be monitored regularly during medication.

Objective To explore the function of the baseline model for end-stage liver disease (MELD) scores,MELD-Na scores and iMELD scores in short-term prognosis in the initial treatment of hepatitis B virus (HBV) related acute-on-chronic liver failure (ACLF) patients.Methods 232 HBV-related ACLF patients who received initial treatment in 302 Military Hospital of China from January 2011 to January 2013 were enrolled in this prospective clinical follow-up.The relationship between the baseline MELD scores,MELD-Na scores,iMELD scores and clinical outcomes were analyzed,and the value of these three models for short term prognosis was assessed.Results Finally the 12-week clinical follow-up was completed in 191 patients,with the completion rate of 82.33％.Eighty-five patients died,with the fatality rate of 44.50％.Compared with the survival group,in non-survival group,the baseline of MELD scores (26.65 ± 7.75 vs.21.19 ± 5.42,t=-5.720,P=0.000),MELD-Na scores (29.16 ± 11.35 vs.21.72 ± 6.33,t=-5.729,P=0.000),iMELD scores (47.19 ± 10.96 vs.38.02 ±7.01,t=-7.011,P=0.000),total bilirubin [TBil (μmol/L):374.3 ± 150.1 vs.305.5 ± 147.1,t=-3.182,P=0.002],creatinine [Cr (μmol/L):110.7 ±90.1 vs.71.1 ± 35.1,t=-4.157,P=0.000] and international normalized ratio (INR:2.3 ± 0.9 vs.2.0 ± 0.6,t=-2.754,P=0.006) were significantly increased,but the baseline of serum Na+ (mmol/L:132.8 ± 6.1 vs.136.7 ± 5.1,t=4.861,P=0.000) was significantly lowered.It was shown by Spearman correlation analysis thai the baseline MELD scores,MELD-Na scores and iMELD scores all had positive correlation with the short-term prognosis of patients (r value was 0.398,0.404,and 0.470,respectively,all P=0.000),the baseline of serum Na+ had a negative correlation with the short-term prognosis of patients (r=-0.365,P=0.000).It was shown by receiver operating characteristic curve (ROC curve) that the cut-off scores of the baseline of MELD scores,MELD-Na scores and iMELD scores were 25.07,25.43 and 43.11 respectively,and the area under ROC curve (AUC) of the baseline of MELD scores,MELD-Na scores and iMELD scores were 0.731,0.735 and 0.773,respectively.The sensitivity of the three models was 55.3％,57.7％,63.5％,and the specificity was 84.9％,84.0％,84.9％ respectively.The value of the three models had no difference in short-term prognostic prediction.According to the respective cut-off score,the three prediction models were divided into four groups,and all of them had differences in fatality rate on the whole (x2 for MELD scores was 34.740,P=0.000; x2 for MELD-Na scores was 36.861,P=0.000; x2 for iMELD scores was 50.127,P=0.000).The mortality was elevated gradually as the equation scores increased.Conclusion The baseline of MELD scores,MELD-Na scores and iMELD scores can predict well the short-term prognosis of the initial treatment in HBV-related ACLF patients,and have relatively good clinical value for guiding therapy.

Objective:To investigate the influence of Covid-19 epidemic on the number of acute ischemic stroke patients with intravenous thrombolytic therapy in Dalian in 2020, the way to hospital, onset to door time (ODT), door to needle time(DNT), onset to needle time(ONT), and the National Institute of Health Stroke Scale (NIHSS) before and after intravenous thrombolytic therapy.Methods:This was a retrospective descriptive study on 13 map-participating hospitals in Dalian from August 2020 to December 2020. The number of acute ischemic stroke patients with intravenous thrombolytic therapy, ODT, DNT, ONT, modes of transport to hospital, the NIHSS score before and after intravenous thrombolytic therapy, etc were analyzed. Data were collected from August 1, 2020 to December 31, 2020, and compared with the baseline data from the same period in 2019.Results:SPSS 22.0 statistical software and Data Analysis ToolPak were used for statistical analysis of the obtained data, and T tests statistic was used for data analysis. In 2020,the number of acute ischemic stroke patients with intravenous thrombolytic therapy increased (681 cases vs 416 cases), and DNT increased [50 minutes vs 45 minutes P = 0.01]. In 2020, 83.55% of patients indicated self-visit as their main mode of transport to the hospital. In 2020, among the self-visit patients with NIHSS score<15 scores,the number of the patients with the NHISS score unchanged or decreased after intravenous thrombolytic therapy was more (562 scores vs. 535 scores, P<0.001), and the patients with NIHSS score ≥15 scores were more willing to call ambulances to hospital. Conclusions:During the Covid-19 epidemic period of 2020, the number of acute ischemic stroke patients receiving intravenous thrombolytic therapy increased, DNT increase; the patients withNIHSS score<15 scores are more willing to choose to visit the hospital using their own transportation and benefit from intravenous thrombolytic therapy. The patients with NIHSS ≥15 scores prefer to call ambulances to hospital.

Objective To observe the changes of brain neurotransmitters in patients with vasovagal syncope (VVS) with encephalofluc-tuogram technology (ET). Methods From August, 2015 to December, 2016, 30 patients with VVS were selected as case group, 30 controls matched with sex and age were selected from the outpatients without syncope. They were detected the function of gamma aminobutyric acid (GABA), glutamate (Glu), 5-hydroxytryptamine (5-HT), acetylcholine (Ach), norepinephrine (NE) and dopamine (DA) with ET. Results There was no significant difference between the two groups in the values of GABA, Glu, 5-HT, Ach and DA (t<1.680, P>0.05), while the values of NE was higher in the case group than in the control group (t=-3.552, P<0.001). Conclusion VVS may be related to the high level of activity of NE in the brain.

OBJECTIVETo compare the reaction of children receiving computer-controlled local anesthetic delivery system (CCLADS) and a conventional syringe, and evaluate the efficacy of anesthesia and the reaction after treatment.

METHODS30 children aged 4 to 9 years old, whose bilateral primary molars in the same jaw present similar lesions with similar treatment were included in the study. These bilateral primary molars were treated under local anesthesia in 2 separate visits with random use of either the CCLADS or conventional syringe. For the first visit, the first group received CCLADS anesthesia, in which buccal infiltration anesthesia was performed for maxillary primary molars, and periodontal ligament anesthesia was performed for mandibular primary molars. The second group received conventional anesthesia, in which buccal infiltration anesthesia was performed for both maxillary and mandibular primary molars. For the second visit, two groups used alternative anesthestic method. Pain behavior was observed and scored by modified-children's hospital of eastern ontario pain scale (MCHEOPS). Pain perception was rated using facial image scale (FIS).

RESULTSThe MCHEOPS and FIS scores in 1 min injection of children with CCLADS were lower than children with conventional syringe (P<0.05). No difference was found in the FIS score in treatment between two injectors (P>0.05).

CONCLUSIONIn local anesthesia, CCLADS can reduce the injection pain caused by conventional syringe to a certain extent.

Anesthesia, Dental ; Anesthesia, Local ; Anesthetics, Local ; Child ; Female ; Humans ; Injections ; Male ; Molar ; Pain Measurement ; Periodontal Ligament ; Syringes ; Tooth, Deciduous

Objective:To explore the possible mechanism of exacerbation of anxiety-like behavior in db/db mice after distal middle cerebral artery occlusion (dMCAO).Methods:The db/db mice was used to establish a type 2 diabetes mellitus model. Meanwhile, heterozygous db/+ mice and C57 wild-type (WT) mice were chosen as double control groups. Then a permanent distal middle cerebral artery occlusion model was employed as an acute ischemic stroke model. The blood glucose levels before and post-dMCAO surgery on day1, day3, and day5 were detected. The brain tissue loss at 35 days after stroke was measured by immunofluorescent staining of MAP2. The open-field test was performed to estimate anxiety-like behavior and general motor and exploring ability of the animals. Axons and myelin were immunostained with non-phosphorylated neurofilaments (SMI32) and myelin basic protein (MBP), respectively, to evaluate differences in white matter integrity in WT, db/+ and db/db mice 35 days after stroke. The correlation between SMI32/MBP and open field test parameters (time in center and corner) was analyzed. Flow cytometry was employed to detect the amount of T cells and B cells, including regulatory T cells (Tregs) in the brain tissue.Results:Blood glucose levels in db/db mice were significantly higher than db/+ mice and WT mice in both sham and dMCAO groups ( P<0.01). There was no significant difference in brain tissue loss 35 days post-stroke among db/db mice, db/+ mice, and WT mice. In the open field test, there were significant differences in the total distance of db/db mice, db/+ and WT mice in the sham and dMCAO groups. Db/db mice shorter than db/+ mice ( P<0.05), WT mice ( P<0.01), and db/+ mice shorter than WT mice ( P<0.05). There were significant time differences in the center among db/db, db/+, and WT mice in sham and dMCAO groups. In both the sham and dMCAO groups, db/db mice spent less time in the center area of the open field than WT mice ( P<0.01). In the sham group, db/+ mice spent less time in the center area than WT mice ( P<0.05). In dMCAO group, db/db mice spent less time in the center area than db/+mice ( P<0.05), and db/+ mice spent less time in the center area than WT mice ( P<0.01). For the time in the corner, in both the sham and dMCAO groups, db/db mice and db/+ mice consumed more time than WT mice ( P<0.01 or <0.05). In the dMCAO group, db/db mice spent more time in the corner than db/+ mice ( P<0.05). Referring to white matter injury, an increased SMI32/MBP ratio in EC area and CTX area (data was not shown in this article) after dMCAO in db/db, db/+ and WT mice were detected. In EC area, db/db mice have a higher SMI32 ratio than db/+ mice and WT mice: 4.24 ± 0.37 vs. 1.96 ± 0.37, 1.80 ± 0.36, both have significant differences ( P<0.01). For db/db mice and WT mice, the SMI32/MBP ratio negatively correlates with time in center and positive correlation with time in the corner. Three days after dMCAO, the total cells of CD 3+ T cells, CD 8+ cells, Tregs, in db/db mice group have significantly decreased compared to WT group: 4 079 ± 1 345 vs. 70 055 ± 3 374, 141.30 ± 28.36 vs. 2 714.00 ± 463.20, 148.00 ± 61.15 vs. 3 007.00 ± 639.90 ( P<0.01), while B cell has no change between two groups. Conclusions:By comparing the severity of anxiety-like behavior of db/db mice, the severity of white matter injury, and the number of T cells and B cells in brain tissue after dMCAO, immune-mediated brain white matter injury may aggravate db/db mice′s post-dMCAO anxiety-like behavior. Due to the gene dose effect, db/+ mice are not suitable as a control group for db/db mice in animal experiments involving anxiety-like behavior assessment.

Objective To evaluate the efficacy of interventional therapy via apical approach in complex mitral valve paravalvular leakage (PVL).Methods From January 2014 to December 2016,interventional therapy by apical approach was used in 7 patients with mitral valve PVL after mechanical valve replacement.There were 5 male and 2 female with mean age of 42-64 (51.3 ± 7.1) years.Six patients had a history of previous infective endocarditis.There were 3 cases of NYHA heart function =Ⅲ,and 4 cases =Ⅳ.The period of time between interventional therapy and previous operation was 6.5-8 (3.6 ± 3.1) years,with mitral regurgitation volume:9.5-23.1 (13.3 ± 4.7)ml.Interventional therapy of small incision method via the left sixth intercostal was carried out in the catheterization laboratory or the hybrid operation room with the patient under general anesthesia.Follow-up evaluation included peri-operational mortality,complications,improvement of cardiac function,hemolysis and postoperative residual mitral regurgitation.Results The success rate of total operation was 100％.The average operation time was 90-300 (145.7 ± 71.8) min,and the DSA radiation time was 6-25 (12.1 ± 6.5)min,with average hospitalization time of 5-12 (10.2 ± 3.5)d.The main post-operative complications included 1 case of hemoptysis,1 case of hematuria and acute renal failure,and 2 cases of blood transfusion,with blood transfusion volume of 1 200 ml and 3 290 ml,respectively.During the follow-up,there was no death.Mitral regurgitation volume decreased to 0 -1.0(0.43 ± 0.45) ml (P ＜ 0.05).All patients had improved heart function in different degree and no serious complications.Conclusions Interventional therapy via apical approach in complex mitral valve PVL has the advantages of being a simpler and safer approach,less trauma,shorter time of hospitalization,faster post-operative recovery and lower treatment cost.Its disadvantages are high difficult operation,strict indications and high technical requirements for the surgeon.By improving operation skills,choosing individualized occluders and treatment of early hemolytic complications,the success rate and long-term effect of interventional therapy can get further improved.

Objective:To investigate the early clinical effects of home-based mobile health intervention on knee joint function after anterior cruciate ligament (ACL) reconstruction.Methods:This was a prospective randomized controlled trial. Patients undergoing arthroscopic ACL reconstruction alone at the Institute of Sports Medicine, Peking University Third Hospital from April 2019 to December 2019 were elected in the trial. Patients were divided into the intervention group and the control group according to random number method. The control group only received the guidance of the traditional paper rehabilitation schedule, while the intervention group also received personalized guidance of electronic rehabilitation prescription and follow-up notice delivered by a WeChat Mini Program Rehabilitation Cloud Platform in addition to the traditional guidance. Joint range of motion (ROM), visual analogue scale (VAS) and thigh circumference difference were assessed before and after the operation. Independent sample t test or Mann-Whitney U test was used for comparison of data between groups, and chi-square test was used for comparison of enumeration data. Results:One hundred and twenty five patients were enrolled in the trial, including 99 males and 26 females, with an average age of 29.0 years (range:18 to 45 years) and an average body mass index of 24.8 kg/m 2 (range:15.8 to 36.6 kg/m 2). At 6 weeks, follow-up was available in 106 cases the target-reach rate of ROM in control group was 42.6%(23/54), which was 67.3%(35/52) in the intervention group, the difference was statistically significant (χ 2=6.53, P=0.01); VAS of the intervention group was 2.5(2.0), lower than that of the control group 3.0(2.0)( M( Q R)), the difference was statistically significant ( Z=-2.06, P=0.04); And the thigh circumference difference of the intervention group was 2.0(2.0)cm lower than that of the control group 3.0(1.8)cm, the difference was statistically significant ( Z=-3.00, P<0.01). Conclusion:Mobile health intervention can improve the early postoperative rehabilitation effect after ACL construction and the ROM of the knee joint, and reduce the bent-knee pain along with the thigh circumference difference.