Objective To assess the prevalence of the A to G mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene in type 2 diabetes in a Chinese population. Methods We screened 716 randomly selected, unrelated patients with type 2 diabetes for the mutation with a PCR-RFLP technique. Results Three individuals with this mutation were identified, representing approximately 0. 4％ of the type 2 patients screened. Further screening of first degree relatives of these 3 patients identified another 4 affected carriers. In comparison with type 2 diabetic patients without the mutation, these 7 carriers of the mt3243 mutation had:①an earlier onset of diabetes (38. 0±10.1 yr vs 53. 4±10.0 yr, P ＜0. 001) ;②lower BMI (19.5±2.0 vs 24. 9±10. 9, P ＜0. 0001) ;and ③ lower post-challenge insulin levels (Area under the curve of insulin levels during the OGTT, 2946± 1647.2 vs 7469±6647.7, P ＜ 0. 01). In addition, we screened the same 716 patients with type 2 diabetes, as well as 181 controls with normal glucose tolerance,for a newly described mt 3316 G→A mutation. This mutation was found in 16 patients with type 2 diabetes (2.20％) and 5 controls (2.7％). Therefore, the frequency of the mutation was not different between patients and controls.Moreover, clinical characteristics such as age of onset of diabetes, BMI, and insulin levels were not different between diabetic patients with the mt3316 mutation and those without it. Concision The mt3316 G→ A mutation is a polymorphism unrelated to diabetes.

Objective: To design qualitative quality control testing device for portable medical endoscope optical performance. Methods:To install the mounting bracket, uniform light source and testing target, etc and integrate testing of the visual resolution, geometric distortion, color reproduction capability, the dynamic range and the brightness of the image plane on only one testing target. Results:This kind of device can observe the target by eye or monitor. Conclusion:It can realize the function of rapid, qualitative detection for clinical medical endoscope optical performance parameters.

Emphasizing practice is a main characteristic in Pharmacy.Insisting on taking employment as guide to do profitable research and practice on new mode of production and practice in pharmacology of senior vocational college from regulating education schedule,establishing and consummating exercitation examination and evaluation mechanism,promoting collaboration between college and corporation,and establishing exercitation-driving education mechanism,can help students improve their practice knowledge in exercitation,transform their knowledge to ability,and cultivate professional pharmacy talent of application and creation.

Based on the gene sequence of AIV matrix protein 2(M2) and two cytotoxic T-lymphocyte epitopes derived from nucleoprotein,the prokaryotic expression vector pET-3M2e-NP1-NP2 was constructed.The target gene was expressed in the solvable form in BL21(DE3) when induced with 1.0 mmol/L IPTG and Western blot analysis showed that the expression product had good immunogencity.Purified fusion protein was mixed with various amount of adjuvant,including Freund's,Vash oil and chitosan,and then immunized 20-day-old chicken by intramuscular injection and boosted 3weeks later.Blood samples were collected weekly following the primary vaccination.The anti-M2e antibody was detected with ELISA method with the synthesized peptide as coated antigen;the neutralizing ability of anti-serum was evaluated on MDCK cell line and chick embryo,the CD4+ and CD8+ T lymphocyte amounts in peripheral blood of immunized chicken was measured by flow cytometry.Results showed that the fusion protein can induce immunological reaction,and the antibody can bind with the viral M2 protein that expressed on the surface of MDCK cells.Flow cytometry result showed that CD4+ and CD8+ T lymphocyte in peripheral blood increased obviously following immunization(P

M2e gene of three copies for H5N1 subtype AIV was synthesized and fused with human mycobacterium tuberculosis hsp70 gene.The fused gene was cloned into the prokaryotic expression vector to get pET-3M2e and pET-3M2e-hsp70.Recombinant protein r3M2e and r3M2e-hsp70 were successfully expressed induced with IPTG and purified with Ni2+-NTA collumn.Following that,the immunity of the recombinant protein was analysized with Western blot.20-day-old AIV non-immunized chickens were vaccined with r3M2e and r3M2e hsp70,at the same time,Trx and KLH-M2e inoculated chickens were served as vector and positive controls.Two weeks after the primary vaccination,every group was boosted with the same vaccine as in the primary vaccination.The humoral immunity of the vaccined chickens was evaluated with antibody detection against M2e,cytopathic suppression test,and indirect fluorescence assay.The cellular immunity was estimated according to lymphocyte subtype analysis with flow cytometry and M2e specific cytokine detection.Four weeks after the boost vaccine,all groups were challenged with 100EID50 AIV of H9N2 subtype,and the virus from swabs was detected with Real-time PCR.Results indicated that r3M2e hsp70 vaccined chicken developed the better humoral and cellular immune response,also,made a better performance compared with r3M2e vaccined group in virus challenge.

Objective:To compare the difference and correlation of HPLC and enzyme-multiplied immunoassay test( EMIT) for the determination of CsA in human whole blood. Methods:A total of 119 clinical samples at different concentrations of CsA were collected and respectively determined by HPLC and EMIT. The difference and correlation of the two determination methods were investigated. Results:There was significant difference in the blood concentrations of CsA determined by HPLC and EMIT(P<0. 05). CsA concen-tration determined by EMIT was 26. 2 ng·ml-1 higher than that determined by HPLC, and 95% CI was (14. 6-37. 7) ng·ml-1 . A satisfactory correlation was achieved between the two methods(r=0. 997 4). Conclusion:There is statistically significant difference in the CsA concentration in whole blood respectively determined by EMIT and HPLC. Attention should be paid to CsA monitoring by E-MIT and HPLC, and relevant adjustment should be carried out.

ObjectiveTo investigate the mechanism of protective effect of Xuebijing injection on vascular endothelial cells in rats with heat stress.Methods Ninety Sprague-Dawley(SD) rats were randomly divided into control, model and Xuebijing injection treatment groups, 30 rats in each group. Heat stress model was reproduced by placing rats in constant temperature box at 40℃, 60% relative humidity for 1 hour, Xuebijing injection group was treated by intraperitoneal injection of Xuebijing 2.5 g/kg, while the control and model groups were treated by intraperitoneal injection of normal saline 2 mL/kg, once a day only in 1 day for both groups. After model establishment, the rectum temperature, heart rate and the mean arterial pressure(MAP) were recorded at 2, 6, 12 hours in each group. At the same time, the rat abdominal aortic blood was collected and serum was separated, the enzyme-linked immunosorbent assay(ELISA) was used to determine the aortic serum levels of lipopolysaccharide(LPS), nuclear factor-κB(NF-κB) and p53, and the prothrombin time(PT), activated partial thromboplastin time(APTT) and D-dimer of venous blood were detected by automatic blood coagulation analyzer(ACLTOP).Results Compared with those in control group, the rectum temperature, heart rate, LPS, NF-κB, p53, PT, APTT, D-dimer were significantly increased, and MAP was obviously decreased in model group(P<0.05 orP<0.01). Compared with model group, the above indexes were improved significantly in Xuebijing injection treatment group at 2 hours〔rectum temperature(℃): 38.02±0.22 vs. 39.32±0.33, heart rate(bpm): 507±14 vs. 562±35, MAP(mmHg, 1 mmHg=0.133 kPa): 98±6 vs. 87±13, LPS(ng/L): 0.65±0.03 vs. 0.82±0.05, NF-κB(ng/L): 1.10±0.04 vs. 1.33±0.05, p53(ng/L): 1.33±0.03 vs. 1.73±0.02, PT(s):15.47±1.03 vs. 20.28±2.01, APTT(s): 40.26±2.46 vs. 47.46±3.51, D-dimer(μg/L): 238.54±8.32 vs. 323.12±8.14,P<0.05 orP<0.01〕.Conclusion Xuebijing injection can correct the disorders of blood PT, APTT, D-dimer via decreasing the secretion of the levels of NF-κB, p53 from vascular endothelial cells in rats with heat stress, thus the integrity of the vascular endothelium can be protected, and LPS entering into the blood stream can be inhibited.

Objective To evaluate the effect of liver function on liver enhancement in hepatobiliary phase of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid(Gd-EOB-DTPA)-enhanced MRI.Methods Sixty-seven patients who suffered from cirrhosis and received enhanced MRI with Gd-EOB-DTPA were retrospectively analyzed,and divided into three subgroups according to ChildPugh score(45 patients in group A,20 in group B,5 in group C).All the individuals of both groups had MRI before injection,and hepatobiliary phase images were obtained at 5,10,and 20 minutes after bolus administration of Gd-EOB-DTPA.The relative enhancement(RE) was calculated by dividing the signal intensity of liver(SI) at t min after injection(SIt) by precontrast SI(SI0).The total serum bilirubin level(TB),serum albumin level(Alb) and prothrombin time(PT) were recorded.The one-way ANOVA was used to compare the RE among three groups at 5,10 and 20 minutes.SNK was used for further pairwise comparison.The effect of liver function on RE was assessed with the generalized linear model.Pearson correlation coefficients were measured between each biochemical test result(TB,Alb,PT) and RE at different time points.Results The RE at 5,10 and 20 minutes were 1.59±0.20,1.65±0.22,1.69±0.25 of group A; 1.47± 0.14,1.48±0.18,1.50±0.22 of group B,1.35±0.07,1.27±0.06,1.26±0.06 of group C.There were statistically significant differences of RE among groups at 5,10 and 20 minutes(F=5.854,11.207,9.666,P＜0.01).Statistically pairwise comparison differences of RE were found between group A and C at 5,10 and 20 minutes(P＜0.01),between B and C at 10 and 20 minutes(P＜0.05),between A and B at 10 minutes(P＜ 0.05).There were statistically significant differences of TB,Alb and PT among groups(P＜0.01).RE at 10 and 20 minutes had moderate negative correlation with TB(r=-0.483,-0.500; P＜0.01),low negative correlation with PT(r=-0.326,-0.351;P＜0.01) and weak positive correlation with Alb(r=0.290,0.292;P＜0.05).Conclusions There are differences of RE among patients with different liver function,and the RE is associated with TB,Alb and PT.Thus,it may allow us to estimate the liver function.

Objective To investigate the feasibility and results of the application of both expanded cutaneous flap and temporoparietal fascia flap in total ear reconstruction with Medpor framework.Methods The main procedures consisted of three stages:stage Ⅰ-skin expansion,stage Ⅱ auricle formation that consisted of orientation of Medpor implant and creation of coverage for the implant by both expaned skin flap and temporoparietal fascia flap,and stage Ⅲ-treatment of residual ear and recontruction of auricular lobule. Results Twenty-two ears in 22 unilateral microtia patients were constructed using Medpor implants covered with both expanded cutaneous flap and temporoparietal fascia flap over the last three years,and they were accepted as pleasing by the patients.Conclusions Application of both expanded cutaneous flap and temporoparietal fascia flap can assure no extrusion of Medpor implant in ear reconstruction.Further more,the two layers of transferred tissues will not affect the profile details of the reconstructed ear.And because the skin covering the framework and fascia is derived from frommastoid region,the appearance and profile of the reconstructed auricle is true to nature and close to that of the opposite one.

Objective To investigate the satlsfaction of senior outpatients over 60 years old,and explore the factors associated with their satisfaction so as to provide scientific references for improving quality of care at the hospital Methods A questionnaire survey was made by random sampling at the hospital.The outcomes subject to Ordinal regress Results The general satisfaction was as high as 82.77％ among these outpatients.Results of the Ordinal regression model showed seven factors significantly affecting such satisfaction rate,namely waiting time for registration,correct statement of medicine information,patience to hear patient's concerns,carefulness of inspection,time for doctor-patient communication,request for comments on therapy,and outpatient clinic seating conditions.Conclusion Senior outpatients tend to emphasize medical skills and medical service than waiting time at the hospital.This suggests better medical skills and service attitude as key to higher general satisfaction rate of senior outpatients.