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 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.

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 of Complex Oral Motor Scoring (COMS) and diadochokinetic rate (DR) on evaluation of oral motor dysfunction and speech disorders in children with cerebral palsy. Methods 107 children with cerebral palsy were tested with Chinese-version Articulation Test, Simple Oral Motor Scoring (SOMS), COMS and DR. The correlation among SOMS, COMS and DR was analysesd. Results 85 children were abnormal in SOMS and 94 in COMS. Oral dyskinesia was found in all the children with spastic quadriplegia, dyskinetic, ataxia and mixed type of cerebral palsy, and less in the type of spastic diplegia and hemiplegia. DR was the least in spastic quadriplegia, dyskinetic, mixed type of cerebral palsy, more in the type of ataxia and spastic diplegia, and the most in hemiplegia. The coefficient of correlation among the SOMS, COMS, the longest pronunciation and DR were more than 0.8 (P<0.01). Conclusion The incidence and severity of oral dyskinesia and dysarthria are associated with the types of cerebral palsy. There is good correlation among SOMS, COMS, longest pronunciation and DR.

Background Corneal blindness is one of the major blinding eye diseases in China.With the development and progress of tissue engineering technology,tissue-engineered corneas offers a new approach to the treatment of corneal diseases.To select and cultivate ideal seed cells is a foundation of construction of tissueengineered corneas.Objective This study was to evaluate the efficiency of stripe off the Descemet membrane with endothelium plus enzymic digestion in the isolation of corneal endothelial cells and analyze the bionomics of rabbit corneal endothelial cells (CECs) in vitro.Methods Descemet membrane was stripped from fresh cornea of New Zealand rabbit under the dissection microscope.Descemet membrane with endothelium was incubated in trypsin and EDTA solution at 37 ℃ and then purified for CECs subculture in vitro.The morphology of the cultured cells was observed under the inverted microscope and marked by CM-Dil dye solution.Then the shape of the cells was observed by hematoxylin and eosin staining and the cells were identified for the expression of neuron specific enolase (NSE) using immunochemistry.The viability of the cells were evaluated by trypan blue staining.The surface structure of the cells were examined under the scanning electron microscope.Intercellular zonula occludens-1 (ZO-1) was identified by immunofluorecsence staining.Results A large number of purified CECs were obtained from Descemet membrane with endothelium through enzymic digestion.Cultured cells grew well and formed monolayer 5-7 days later with the cobblestone stone-like arrangement.The survival rate of the cells was 95％.CECs presented with the red annular fluorescence for CM-Dil with the labeling rate ＞90％.NSE was positively expressed in the cytoplasm.Polygon CECs were seen by hematoxylin and eosin staining and showed the brown staining.Abundant microvilli on the cellular surface and interconnected foot process were seen under the scanning electron microscope.ZO-1 showed the green fluorescence.Conclusions The method of striping off the corneal Descemet membrane with endothelium plus enzymic digestion can obtain abundant CECs.Cultured cells have good biological properties.This study may offer a feasible application in the engineering of corneal transplant membrane.

Background The characters of diabetic retinopathy (DR) include capillary occlusion,microcirculation disturbance and retina neovascularization near ischemic areas of the retina.The pathogenic mechanism is still incompletely clear,but the primary mechanism of DR is the change of permeability of retinal blood capillary.Objective The aim of this study was to investigate the effect of lycium bararum polysaccharides (LBP) on the blood glucose,blood fat and permeability of blood-retinal barrier in diabetic rats.Methods Fifty-four clean adult SD rats were randomized into 4 groups.The diabetes mellitus models were created in 36 SD rats by injection of 45 mg/kg streptozotocin(STZ) via caudal vein.The rats with the blood glucose ＞ 16.7 mol/L were defined as the diabetic mellitus in 72 hours after injection.LBP of 200 rmg/( kg · d) was intragastriccally administered in 18 models once a day in LBP group,and equal volume of normal sodium was used at the same way in 18 models in DM group,and 18 normal rats were as normal control group.The rats were killed in 4,8 and 12 weeks respectively for the detect of blood glucose level,triglyceride,total cholesterol.The retina tissue were isolated for the quantification of vascular permeability using Evans blue method.Results The blood glucose level was significantly elevated in the DM group and LBP group compared with normal control group,and that in LBP group was obviously declined in comparison with the DM group with a 57％,40％,36％ fall off in 4,8,12 weeks respectively,showing a statistically significant differences( P＜0.05 ).The triglyceride concentration was considerably raised in the DM group compared with normal control group in all of the time points( P＜0.01 ),however,the rise of triglyceride concentration was found only in 12 weeks in LBP group(P＜0.05).The cholesterol level was increased in 8 and 12 weeks in DM group compared with normal control group and that of LBP group went up only in 12 weeks (P＜0.0l ).The Evans blue contents in dry retina tissue were(26.23±2.00),(29.78± 1.78 ) and( 34.08±3.03 ) μg/g in 4,8,12 weeks in the DM group,and those of LBP were ( 13.27 ±0.77 ),( 18.01 ± 1.77 ),( 25.05 ±:1.50 ) μg/g,showing statistical differences between two groups ( P＜0.01 ).However,in comparison with normal control group ( 12.34 ±4.30,12.76 ± 2.11,12.45 ±4.40 μg/g),the Even blue content were elevated in various time points (P＜0.01 ).Conclusions LBP can suppress the increase of the blood-retinal barrier permeability in STZ-induced diabetic rat by lowing the levels of blood glucose,triglyceride and total cholesterol.These results suggest that LBP has a therapeutical effect on DR.

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.

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.