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.

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.

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.

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 observe the efficacy and safety of sorafenib in the treatment of the unresectable hepatocellular carcinoma.Methods According to the inclusion criteria,33 patients with the unresectable hepatocellular carcinoma were given sorafenib (400 mg for twice per day).During the course of treatment,dose was adjusted based on the degree of the adverse effects.Tumor response to sorafenib and safety was assessed every 6-8 weeks using the modified RECIST criteria.The survial curve for the time to progression (TTP) and overall survival (OS) were estimated.Results In this series,there was no patients achieved complete response (CR) and partial response (PR),1 1 patients were evaluated as with stable disease(SD),22 patients were with progressive disease (PD).The median TTP was 5.6 months (2.3-8.9 months).The median TTP was longer in patients with BCLC B than BCLC C stage.TTP was longer in good than in poor performance status patients,and shorter in extrahepatic metastasis than in no extrahepatic metastasis patients.The overall incidence of adverse events was 75.8％.The most common adverse events were hand foot skin reaction,diarrhea,hypertension and rash.Three patients had grade 3 adverse events.Conclusions Sorafenib can extend the median time to progression in patients with unresectable hepatocellular carcinoma.Patients with earlier stage of HCC and better performance status are hopeful for more positive response to the treatment of sorafenib.

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 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.

Objective To explore the effectiveness of ω-3 fatty acids in intervening rat models of chronic obstructive pulmonery disease (COPD).Methods The rat COPD models were established by cigarette smoking and intratracheal lipopolysaccharide instillation.Totally 90 rats were randomly divided into 3 groups:normal control group (treated with normal saline),COPD group,and intervention group (the COPD rat models treated with ω-3 fatty acids).Lung tissues were obtained on the 7th,21st,and 28th day.The left lower lobes were analyzed to determine the expressions of nuclear factor-kappa beta (NF-κB) and interferon-γ (IFN-γ)and the right lung lobes were sliced for detecting the cell apoptosis.Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) was used to detect the serum IFN-γ and interleukin-6 (IL-6).Results (1) The pathological changes of lung tissue:there were a large number of inflammatory exudation,alveolar wall thickening,hyperplasia of vascular smooth muscle and the alveolar structure destruction in the COPD model group,but the lung tissue were part of alveolar cavity and a little inflammatory exudate in ω-3 fatty fish acids treatment group,control rats were almost no alveolar inflammation on the 28th days.(2) On the 28th day,NF-κB protein expression of the lung tissue (18.91 ± 3.07) in rats of COPD model group was significantly higher than the control group and the intervention group (5.47 ±4.86 and 7.23 ±2.21) (P ＜0.01).On the 28th day,IFN-γ protein expression in lung tissue of the rats in the model group was 7.12 ±3.37,significantly lower than the intervention group (18.74 ± 2.65),the difference was statistically significant (P ＜ 0.01).(3) The IL-6 levels of the blood-serum of model group rats were (13.43 ± 2.47) ng/L,significantly higher than the control group and the intervention group [(4.78 ± 1.93) and (4.98 ± 1.89) ng/L],the differences were statistically significant (P ＜ 0.01) on the 28th day,,but the IFN-γ level [(2.23 ± 0.63) ng/L] in COPD group was more poorer than ω-3 fatty acids group and the intervention group [(4.51 ± 0.71) and (7.05 ± 0.52) ng/L] (P ＜ 0.01).Conclusions The ω-3 fatty acids can lower NF-κB protein expressions in lung tissues and serum and IL-6 levels in COPD rats; aslo,it can increase the IFN-γ protein expression in lung tissue and serum.Thus,it can prevent the lung inflammation in COPD rats.