Objective: To explore influence of exercise on cardiac autonomic nerve function in patients undergoing coronary artery bypass grafting (CABG) and its clinical significance. Methods: A total of 53 patients with coronary heart disease undergoing CABG were randomly divided into routine treatment group (n=25) and rehabilitation group (n=28, received exercise training based on routine treatment). Changes of autonomic nerve function before, second and eighth week after CABG were analyzed in two groups using time domain indexes of heart rate variability (HRV) by ambulatory electrocardiography, including standard deviation of normal to normal RR intervals calculated over the 24 h period (SDNN), standard deviation of normal to normal RR intervals in all 5 min segments of the entire recording (SDANN), root-mean square of differences between successive normal to normal intervals (rMSSD) and adjacent normal RR interval difference > 50ms stroke accounted for a percentage of 24h total RR interval (PNN50). Results: Compared with before CABG, there were significant decrease in SDNN, SDANN, RMSSD and PNN50 in both groups two weeks after CABG (P<0.05~0.01); eight weeks after CABG, above indexes recovered to levels before CABG in routine treatment group[SDNN （113.6±29.4）vs.（116.7±24.7）, SDANN（112.2±32.1）vs.（113.6±28.6）, rMSSD（21.9±8.2） vs.（23.2±7.1）, and PNN50 （7.5±4.2）vs.（8.2±3.7）] , P>0.05 all; Compared with before CABG, there were significant improvements in SDNN [(114.7±25.2) ms vs. (132.6±30.6) ms], SDANN [(111.6±23.5) ms vs. (129.2±30.8) ms], rMSSD [(24.2±8.7) ms vs. (29.9±7.5) ms] and PNN50 [(7.8±2.2) ms vs. (9.5±2.3) ms], and there were significant improvement than those of routine treatment group [SDNN （132.6±30.6）vs.（113.6±29.4）, SDANN（129.2±30.8）vs.（112.2±32.1）, rMSSD（29.9±7.5）vs.（21.9±8.2）and PNN50 （9.5±2.3）vs.（7.5±4.2）] eight weeks after CABG in rehabilitation group, P<0.05 all. Conclusion: All HRV indexes significantly decrease on two weeks after CABG in both groups, suggesting that CABG can damage cardiac autonomic nerve system. These indexes of rehabilitation group were more improvement than those of routine treatment group, suggesting exercise training can more significantly improve cardiac autonomic nerve function after CABG.

Objective To observe hepatitis B virus (HBV) reactivation in 12 patients with rheumatic disease undergoing immunosuppressive therapy and to evaluate whether preemptive antiviral therapy is necessary for patients receiving disease-modifying anti-rheumatic drugs (DMARDs).Methods From January 2008 to March 2012,a total of 12 HBV-infected patients with rheumatic diseases were consecutively enrolled into this long-term follow-up study.Liver function and serum levels of HBV DNA were tested during the follow-up.Results The medium duration of follow-up was 41 months (range 16-48).Four patients received steroid treatment,and among them two patients without pre-emptive antiviral therapy developed HBV reactivation.After administr-ation of LAM or ETV,HBV replication was controlled in both patients.Five patients were treated with disease-modifying anti-rheumatic drugs and the other three patients received tumor necrosis factor-alpha-blocking agents.None of these patients received pre-emptive antiviral therapy.HBV reactivation did not occur in any of them.Conclusion HBV reactivation does occur in HBV-infected patients with rheumatoid diseases after immunosuppressive therapy.Pre-emptive antiviral therapy should be administered in patients who are receiving steroid therapy for rheumatic diseases.In contrast,DMARDs and TNFBA are relatively safe for HBV-infected patients with rheumatic diseases.Close monitoring of HBV DNA and ALT levels is necessary to the mana-gement of HBV reactivation.

Objective To establish a cell line stably expressing the human CD36 by using TA clo-ning and cell transfection technology and to analyze its application to the detection of anti-CD36 antibodies. Methods Total RNA was isolated from human platelets and then used to synthesize complementary DNA ( cDNA) . Sequence of the gene encoding CD36 on human platelets was obtained by PCR amplification. The recombinant vector was transformed into TOP10 E. coli after TA cloning. The positive recombinant pcDNA3. 1/V5-CD36 plasmid was screened out by blue-white selection and then sequenced. The correctly constructed plasmid coated with Effectene? Transfection Reagent was transferred into HEK293T cells. Fluo-rescence-activated cell sorting was performed to screen out the cell line that could stably express the CD36 on human platelets. The transfected cell line-based flow cytometry analysis and antibody capture assay ( ACA) were established and used for antibody detection in nine serum samples positive for anti-CD36 antibodies. Results The HEK293T cell line stably expressing the recombinant CD36 was successfully established. Compare with the monoclonal antibody immobilization of platelet antigens assay ( MAIPA) , anti-CD36 anti-bodies could be easily identified in nine serum samples by using the transfected cell line-based flow cytome-try analysis and ACA. Conclusion This study suggests that the HEK293T cells stably expressing the re-combinant CD36 could be used in flow cytometry analysis and ACA for the detection of anti-CD36 antibod-ies. It also paves the way for further researches on the mechanism of CD36 in other diseases.

Objective To evaluate the changes in the expression of c-fos protein in the spinal cord in a rat model of oxycodone dependence or withdrawal response.Methods Thirty SPF adult male Sprague-Dawley rats,aged 6-8 weeks,weighing 180-220 g,were divided into 3 groups (n=10 each) using a random number table method:normal saline group (group NS),oxycodone dependence group (group OD),and oxycodone withdrawal group (group OW).In OD and OW groups,oxycodone was injected subcutaneously in back,5 days in total,with the dose of 2,3,4,5 and 6 mg/kg in turn,3 times a day (8:00/15:00/22:00).The equal volume of normal saline was given instead in group NS.The mechanical paw withdrawal threshold was measured at 3 days before administration and 30 min after the last administration every day.The oxycodone withdrawal was induced by intraperitoneal injection of naloxone 4 mg/kg at 8 h after the last administration of oxycodone on 5th day in group OW.The withdrawal response scores and range of weight changes were recorded within 15 min after giving naloxone or normal saline in NS and OW groups.Spinal cord tissues were collected at 1 h after the last administration on 5th day in group OD and at 1 h after giving normal saline or naloxone on 5th day in NS and OW groups for determination of the expression of c-fos protein by Western blot.Results Compared with group NS,the mechanical paw withdrawal threshold was significantly increased on 1 and 2 days after administration,and the expression of c-fos protein in the spinal cord was up-regulated in OD and OW groups,and withdrawal response scores were significantly increased,and the range of weight change was increased in group OW (P＜0.05).The expression of c-fos protein was significantly down-regulated in group OW as compared with group OD (P＜0.05).Conclusion Oxycodone dependence or withdrawal response may be related to the expression of c-fos protein in the spinal cord of rats,and the expression is up-regulated during oxycodone dependence,while down-regulated during oxycodone withdrawal.

OBJECTIVE: To construct eukaryotic expression vectors for human platelet CD36 gene 220 C>T and 429+4insg variants and analyze their expressions in HEK293T cells. METHODS: RNA was isolated from human platelets and reversely transcribed into cDNA. Sequences of 220C>T and 429+4insg variants were derived by PCR amplification. The target sequence was ligated into a pcDNA3.1/V5-His-TOPO vector by TA cloning, which was transformed into TOP10 E. coli. Positive plasmids were screened by blue-white selection. After sequencing, plasmid DNA carrying 220C>T or 429+4insg variant was used to transfect HEK293T cells with the help of effectene. Expression of CD36 protein was then analyzed by flow cytometry and Western blotting. RESULTS: An eukaryotic expression vector pcDNA3.1/V5-His-CD36 (220C>T/429+4insg) was constructed by TA cloning. After transfected into HEK293T cells, the 220C>T and 429+4insg variants resulted in CD36 deficiency in HEK cells, which was confirmed by flow cytometry and Western blotting. CONCLUSION The 220C>T and 429+4insg variants can cause CD36 deficiency in human platelets. This system may be used for assessing the effect of 220C>T, 429+4insg, and other variants on the expression of CD36.
Blood Platelets ; CD36 Antigens ; Cloning, Molecular ; Escherichia coli ; Eukaryota ; Genetic Vectors ; HEK293 Cells ; Humans ; Plasmids ; Transfection

Objective To develop a solid-phase red cell adherence(SPRC A)reagent kit for unexpected alloantibody i-dentification.Methods A solid-phase carrier for bounding red blood cell(RBC)was prepared by coating microstrip with succinimide,then panel cell suspensions were dispensed to microwells to produce RBC monolayers,after that,1×PBS(pH7.40)was used to lysis the RBC monolayer to form stroma monolayer.Prior to the drying procedure,drying medium suspension was added to microwell to protect stroma monolayer.The solid-phase microplate was successfully produced after the stroma had been dried in refrigerator at 4℃.IgG anti-E and anti-Fya with low titer were tested every two months with the reagent kit to observe its stability.Seventy-two plasma samples containing unexpected antibodies and 152 normal plasma samples were determined with SPRCA reagent kit and column agglutination technique(CAT).Performances including sensi-tivity and specificity of the two reagents were compared by paired chi-square test.Results RBC stroma monolayer could be attached well on microwell bottom coated with succinimide.Drying medium suspension could provide effective protection for stroma monolayer.Both IgG anti-E and anti-Fya with low titer were determined well by the reagent kit during the storage peri-od.Of the 72 positive samples,70 were identified correctly by SPRCA kit and 71 by CAT[Sensitivity:97.22％(70/72)vs 98.61％(71/72)].Of the 152 negative plasma samples,149 were correctly detected by SPRCA kit and 152 by CAT[Spe-cificity:98.02％(149/152)vs 100％(152/152)].Paired chi-square test showed no significant differences between the two reagents(P＞0.05).Conclusion A SPRCA reagent kit for IgG unexpected antibody identification had been developed suc-cessfully.

Objective To investigate the association between types of obesity and the 10-year-coronary heart disease risk in Tibet and Xinjiang of China.Methods Using the multi-stage random sampling method,7 631 participants aged 35 or older were examined under the International Standardized Examination process but with only 5 802 were eligible for analysis,in the 2015-2016 season.Results The prevalence rates of general obesity,central obesity,visceral obesity and compound obesity were 0.53％,12.62％,10.08％ and 42.35％,respectively.Out of all the compound obesity cases,58.65％ (1 441/2 457) of them appeared as having all types of obesity in our study.Risk related to the 10-year-coronary heart disease was higher in men than in women [(3.05 ± 4.14)％ vs.(1.42-2.37) ％,P＜0.000 1.Compound obesity (30.16％) showed the highest proportion on the risk of 10-year-coronary heart disease than central obesity (28.01％),visceral obesity (18.46％) or the general obesity (19.35％).After adjustment for confounding factors,results from the multivariate analysis showed the risk in compound obesity was higher than central obesity,visceral obesity or general obesity and was associated with the highest risk on the 10-year-coronary heart disease (OR=2.889,95％CI:2.525-3.305).People with anomalous BMI and WC seemed to have had the higher risk (OR=3.168,95％CI:2.730-3.677).Conclusions Obesity was popular in the residents of Tibet and Xinjiang areas of China.Men and people with compound obesity (especially both BMI and WC were abnormal) seemed to carry greater risk on the 10-year-coronary heart disease.

Objective To investigate the association between body fat percentage (BFP),visceral fat index (VFI) and Cardiometabolic Risk Factor Clustering (CRFC),among population aged 35 or older in Tibet and Xinjiang areas.Methods Using the stratified multi-stage random sampling method,7 571 residents aged 35 or above were examined with international standardized examination between 2015 and 2016.Of the eligible 5 643 participants,association of BFP and VFI with CRFC was defined as having two or more of the four risk factors:hypertension,diabetes mellitus,high TG and low HDL-C,at the same time.Logistic regression analysis and receiver operating characteristic (ROC) curve analysis were employed to further explore the relationships.Results The overall prevalence of CRFC among aged 35 and older population in Tibet and Xinjiang areas was 9.78％.BFP and VFI were divided into four groups by quartile.After adjustment for age,gender,race,cigarette smoking,alcohol consumption,education attainments,and altitude of residence,ORs of CRFC seemed to have increased with BFP and VFI.Compared with people having BFP of 5.0％-27.0％,the OR(95％CI) were 1.15(0.86-1.54),1.48(1.05-2.07) and 1.72(1.10-2.68) for the ones who presented 27.1％-31.7％,31.8％-36.6％ and 36.7％-50.0％ of BFP.Compared to people of having 1-6 of VFI,with OR (95％CI) as 1.20(0.81-1.79),1.91(1.30-2.80) and 3.91(2.64-5.77) for the ones having 7-9,10-13 and 14-30 of VFI.Areas under the curve (AUC) of CRFC appeared as 0.55 for BFP and 0.70 for VFI,respectively,with statistically significant difference (P＜0.01).Conclusion Both BFP and VFI levels were closely associated with CRFC while VFI seemed to have a better predictive value than the BFP.

【Objective】 To find the HLA-matched platelets from platelet donor registry and track the transfusion effect for aplastic anemia patients in pregnancy with platelet transfusion refractory (PTR) caused by anti-HLA, so as to support the childbirth and follow-up treatment of the patients. 【Methods】 Antibodies to HPA and HLA were detected by ELISA kit and Luminex, respectively. DNA of the patient and 523 platelet donors from the donor registry were extracted for high-resolution HLA genotyping. The Risk Factors Evaluation Software of PTR was used to select the ABO-compatible and HLA-matched donors, without HLA Eplets specific to the patient. After MASPAT cross matching, the patient was transfused with 1 U of platelets, and the 24h post-transfusion effect was recorded. 【Results】 Only anti-HLAⅠantibody was found in the patient serum, and the specificity Eplet was 65QIA, including HLA-B^*27∶08, B^*27∶05, B^*07∶02, B^*55∶01, B^*67∶01 and B^*54∶01; anti-HLA Ⅱ antibody was negative. The HLA genotypes of both the patient and donor were HLA-A^*02∶07, A^*11∶01, B^*46∶01, B^*46∶01, C^*01∶02, 01∶03, DRB1^*04∶05, DRB1^*0901, DQB1^*03∶03 and DQB1^*04∶01. The results of MASPAT matching were negative. HLA-matched platelets transfusion provided a satisfactory posttransfusion platelet responses in patients(1 before vs 33 ×10⁹/L after). A baby boy was delivered by cesarean section 4 weeks later, and the same donor was recruited due to the mother′s low Plt and bleeding trend. The 24h posttransfusion Plt (×10⁹ / L) rose from 5 to 37 after the secondary transfusion of 1U platelet. The vital signs of the mother and her baby were normal during the two-day follow up. 【Conclusion】 The establishment of blood donor registry and screening of HLA-matched donors is an effective approach to treat PTR caused by HLA antibodies in pregnancy complicated with aplastic anemia.

Nanotechnology has emerged as an ideal approach for achieving the efficient chemo agent delivery. However, the potential toxicity and unclear internal metabolism of most nano-carriers was still a major obstacle for the clinical application. Herein, a novel "core‒shell" co-assembly carrier-free nanosystem was constructed based on natural sources of ursolic acid (UA) and polyphenol (EGCG) with the EpCAM-aptamer modification for hepatocellular carcinoma (HCC) synergistic treatment. As the nature products derived from food-plant, UA and EGCG had good anticancer activities and low toxicity. With the simple and "green" method, the nanodrugs had the advantages of good stability, pH-responsive and strong penetration of tumor tissues, which was expected to increase tumor cellular uptake, long circulation and effectively avoid the potential defects of traditional carriers. The nanocomplex exhibited the low cytotoxicity in the normal cells