This paper reported a new home-made nonpoisonous substrate of horseradish peroxidase(HRP),3,3′,5,5′-tetramethylbenzidine sulphate(TMBS),used in ELISA with the aqueous soluble egg antigen(SEA)as antigen;proved patients'sera as antibo- dy and the antihuman IgG was replaced by protein A conjugate. The optimun condition obtained by orthogonal design was 1.0 millimolarity of TMBS,1.2 millimolarity of H_2O_2,and pH value at 5.0.The best time to stop reaction with H_2SO_4,was 30-60 min after adding substrate TMBS.At the same time,TMB and OPD were used as compared substrates.It showed that the reaction colour of both TMBS and TMB remained much more stable than that of OPD when being exposured in light and oxygen.After the solutions of TMBS and TMB were stored under 4℃ in refrigerator for 60 days,the coefficient of variation among different experiments was less than 10%. The results in detecting specific antibody in sera from 28 patients with proved schistosomiasis also showed that the sensitivity of TMBS was remarkably higher than OPD.It was found that TMBS used in enzyme immunoassay for demonstrating specific antibody was suitable not only for optical density(OD)read by spectrophotometer but also for estimation judged by naked-eye.During TMBS and TMB used in indirect immunoperoxidase method(IIP),the specific antibody in the tissue can be easily dete- cted and located as well.

To explore the effect of quercetin on the proliferation and apoptosis of HeLa cells, HeLa cells were incubated with quercetin at different concentrations. Cell viability was evaluated by MTT assay, cell apoptosis was detected by Annexin-V/PI double labeled cytometry and DNA ladder assay. Cell cycle was flow cytometrically determined and the morphological changes of the cells were observed under a fluorescence microscope after Hoechst 33258 staining and the apoptosis-related proteins in the HeLa cells were assessed by Western blotting. The results showed that quercetin significantly inhibited the growth of HeLa cells and induced obvious apoptosis in vitro in a time- and dose-dependent manner. Moreover, quercetin induced apoptosis of HeLa cells in cell cycle-dependent manner because quercetin could induce arrest of HeLa cells at G0/G1 phase. Quercetin treatment down-regulated the expression of the PI3K and p-Akt. In addition, quercetin could down-regulate expression of bcl-2, up-regulate Bax, but exerted no effect on the overall expression of Akt. We are led to conclude that quercetin induces apoptosis via PI3k/Akt pathways, and quercetin has potential to be used as an anti-tumor agent against human cervix cancer.

Food Sanitation Law which was issued for many years has no longer been congruent with the current development of socio-economy. Under this situation,Food Safety Law came into being. Although the promulgation of the law is important,the task of carrying out the law is more difficult. On the basis of concluding the problems in carrying out the food sanitation law and the analysis of its causes,it is significant to discuss the Food Safety Law from the viewpoint of Food Sanitation Law enforcement.

Objective: To investigate the effects of preconditioning with lithium on the expressions of HSP70 in gerbil after global ischemia. Methods: The transient ischemia model of gerbil global was established by clamping the bilateral common carotid arteries for 5 min.36 gerbils were randomly divided into group lithium(n=18) and group control(n=18).Gerbils in group lithium were injected with lithium intraperitoneally,5 mmol/kg,once a day for 5 consecutive days.Normal saline was used instead of lithium in group control as vehicle control.Both group lithium and control underwent ischemia after preconditioning,and were then further divided into three subgroups(n=6,in each subgroup).The garbles in lithium 1 or control 1 subgroup were sacrificed 1 day after ischemi;lithium 3 or control 3 sacrificed in 3 days;and lithium 7 or control 7 were sacrificed in 7 days.The brains were made into paraffin sections and were used for immunohistochemistry against HSP70. Results: Compared with the vehicle control,the HSP70 expression in subgroup lithium1 was inhibited in the hippocampus CA1(P

Objective To study the advantage and disadvantage of two types of rat heart trans- plantation models.Methods Twenty Wistar rats served as donors and 20 SD rats as recipients ran- domly in two groups.Abdominal working and nonworking cardiac transplantation models from Wistar to SD rats were established.In the working model,the donors pulmonary artery was anastomosed to the left atrial appendage.The left ventricle ejected volume through an end to side anastomosis of the donor's aorta to the recipient's abdominal aorta.In the nonworking cardiac transplantation model,the donor's pulmonary artery was anastomosed to the recipient's inferior vena cava and the donor aorta was anastomosed to the recipient aorta.Results Working model and nonworking model were successfully established.The survival rate of both models was 90 %.Total operating time of the working model was 10 min shorter than the nonworking model.Postoperative UCG demonstrated the working trans- planted hearts had ejection.Conclusions Rat abdominal working cardiac transplantation model was used perfectly with high rate of survival and shorter total operating time.The working model approached cardiac physiological functions more than the nonworking model.

Objective To evaluate the clinical effect of emergency pelvic external fixation in hemodynamically unstable patients with pelvic fracture.Methods The clinical data of 56 hemodynamically unstable patients with pelvic fracture from January 2008 to December 2013 were selected.The changes of hemoglobin,mean arterial pressure,heart rate,serum lactate,urine volume,norepinephrine dosage before and after surgery for 24 h were compared and analyzed.Results Compared with those before surgery,the levels of hemoglobin,mean arterial pressure,serum lactate,urine volume,heart rate after surgery for 24 h were significantly improved:(109 ± 12) g/L vs.(69 ±22) g/L,(87.8 ±6.5) mmHg (1 mmHg =0.133 kPa) vs.(55.8 ± 7.7) mmHg,(6.3 ± 5.1) mmol/L vs.(10.8 ± 6.6) mmol/L,(35.9 ± 9.9) ml/h vs.(17.6 ± 8.4) ml/h,(84.2 ± 15.4) times/min vs.(146.4 ± 12.1) times/min,and the norepinephrine dosage was significantly reduced:(0.8 ± 0.2) μ g/ (kg· min) vs.(2.2 ± 0.8) μ g/ (kg· min).The differences had statistical significance (P ＜ 0.05 or ＜ 0.01).Conclusion Bedside pelvic external fixation can rapidly improve shock symptom in hemodynamically unstable patients with pelvic fracture,which is safe and efficient.

Objective To investigate instant messenger management on moderate asthma control in young and middle-aged patients.Methods Sixty young and middle-aged asthma patients enrolled were divided into 2 groups by random number table,traditional management group (group A) and messenger management group (group B),and received the asthma treatment with traditional management or messenger management for 1 year.The compliance,control condition,pulmonary function and Saint George respiratory questionnaire(SGRQ) were compared between two groups.Results After one year management,there were 20 patients in proceeding of the standard treatment,15 total control,10 partial control and 5 uncontrolled in group A,while there were 26 patients in proceeding of the standard treatment,21 total control,7 partial control and 2 uncontrolled in group B.Compared with group A,the treatment adherence(x2 =6.331,P ＜ 0.05),satisfaction degree (t =6.051,P ＜ 0.01),asthma control rate (x2 =6.490,P ＜ 0.05),efficacy satisfaction (t =6.051,P ＜ 0.01) were all increased in group B.Forced expiratory volume in 1 second was elevated more significantly in group B than that in group A [(0.132 ± 0.175) L vs.(0.326 ± 0.125) L] (t =5.099,P＜ 0.01).SGRQ scores were decreased more obviously in group B than that in group A (t =4.316,P ＜ 0.01).Conclusions The asthma control,respiratory and pulmonary function can be improved by messenger management in young and middle-aged patients.So promoting the messenger management model actively is advantageous to improve the level of prevention and treatment of asthma in China.

Objective: By observing the differences in Western Ontario and McMaster Universities osteoarthritis index (WOMAC) and visual analog scale (VAS) scores in moxibustion treatment for moderate-to-severe primary knee osteoarthritis (KOA) with moxa of different storage years (3-year moxa and 1-year moxa from Qichun, Huanggang City, Hubei Province, China) through a randomized clinical trial, to objectively evaluate the differences in therapeutic efficacy of moxibustion with moxa of different storage years. Methods: A total of 63 patients with moderate-to-severe KOA who met the inclusion criteria were randomized into moxibustion group 1 and moxibustion group 2 by central randomization method, with 32 cases in moxibustion group 1 and 31 cases in moxibustion group 2. Moxibustion group 1 was treated with moxa stored for 3 years, and moxibustion group 2 was treated with moxa stored for 1 year. Dubi (ST 35), Neixiyan (EX-LE 4) and Heding (EX-LE 2) were selected in both groups, and the treatment lasted 20 min per time, 3 times a week. The immediate efficacy was compared after 6 times of treatment, and long-term efficacy was compared at follow-up 4 weeks after the end of treatment. Results: During the treatment, there were 2 dropouts in moxibustion group 1, and 1 dropout in moxibustion group 2. The total effective rate in the two groups was 83.3％ and 60.0％, respectively. Followed up at 4 weeks after the end of treatment, the total effective rate in the two groups was 80.0％ and 66.7％, respectively. There were no statistical differences between the two groups (both P>0.05). After treatment and 4 weeks after the end of treatment, the WOMAC and VAS scores in both groups decreased significantly compared with those before treatment (all P<0.01); the scores of stiffness item of WOMAC in moxibustion group 1 were lower than those in moxibustion group 2 (both P<0.05); there were no statistical differences in the scores of pain item and dysfunction item of WOMAC, and VAS scores between the two groups (all P>0.05). Conclusion: Moxibustion with moxa of different storage years (stored for 3 years and 1 year) both can improve the pain, stiffness and motor function in patients with moderate-to-severe KOA. While moxa stored for 3 years has a better therapeutic efficacy in improving stiffness of the knee joint than that stored for 1 year.

Animals ; Binding Sites ; drug effects ; Brain ; metabolism ; Calcium ; metabolism ; Cerebral Cortex ; metabolism ; Cyclic AMP ; metabolism ; Hippocampus ; metabolism ; Insecticides ; toxicity ; Male ; Organothiophosphorus Compounds ; toxicity ; Phosphoramides ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Carotid-femoral pulse wave velocity in hemodialysis patients is influenced by multiple factors.OBJECTIVE: To explore the carotid-femoral pulse wave velocity (PWVcf) changes and related factors in maintenance hemodialysis (MHD) patients.DESIGN, TIME AND SETTING: Non-randomized control experiment was performed at the Third Hospital of Peking University from January to August 2006.PARTICIPANTS: A total of 66 MHD patients were selected from Third Hospital of Peking University.METHODS: General clinical conditions, biochemical indexes, PWVcf and subjective nutritional state were evaluated. MHD patients were divided into two groups according to the state of nutrition evaluated with SGA: non-malnutrition (n=49), malnutrition group (n=17). The PWVcf of two groups was compared, and one-way and multiple regression analyses were performed to explore PWVcf-relatest factors.MAIN OUTCOME MEASURES: PWVcf, blood albumin and parathyroid hormone.RESULTS: PWVcf was significantly positively correlated with age (r= 0.284, P< 0.05), systolic blood pressure (r= 0.468, P<0. 001), and pulse pressure (r=0.451, P< 0.001), while negatively correlated with prealbumin (r=0.318, P< 0. 05),plasma-albumin (r=0.263, P< 0.05), parathyroid hormone (r=0.167, P< 0.05), serum creatinine (r=0.347, P= 0.004)and transferring (r=0.284, P < 0.05) before dialysis. Multiple regression analysis demonstrated that pulse pressure and parathyroid hormone were independently related with PWVcf. The PWVcf was significantly increased in malnutrition group compared with non-malnutrition group (P < 0.05). CONCLUSION: Pulse pressure is the major clinical determinants of arterial stiffness in patients with MHD independent of conventional risk factors for cardiovascular disease; Malnutrition and atherosclerosis are common complications of MHD,showing close relationship. The PWVcf was significantly increased in malnutrition patients.