Construction of Lactobacillus rhamnosus GG particles surface display system / 生物工程学报

To describe a novel particles surface display system which is consisted of gram-positive enhancer matrix (GEM) particles and anchor proteins for bacteria-like particles vaccines, we treated Lactobacillus rhamnosus GG bacteria with 10% heated-TCA for preparing GEM particles, and then identified the harvested GEM particles by electron microscopy, RT-PCR and SDS-PAGE. Meanwhile, Escherichia coli was induced to express hybrid proteins PA3-EGFP and P60-EGFP, and GEM particles were incubated with them. Then binding of anchor proteins were determined by Western blotting, transmission electron microscopy, fluorescence microscopy and spectrofluorometry. GEM particles preserved original size and shape, and proteins and DNA contents of GEM particles were released substantially. The two anchor proteins both had efficiently immobilized on the surface of GEM. GEM particles that were bounded by anchor proteins were brushy. The fluorescence of GEM particles anchoring PA3 was slightly brighter than P60, but the difference was not significant (P>0.05). GEM particles prepared from L. rhamnosus GG have a good binding efficiency with anchor proteins PA3-EGFP and P60-EGFP. Therefore, this novel foreign protein surface display system could be used for bacteria-like particle vaccines.

Effects of Cerebral Microbleeds on the Incidence of Cerebral Events in Patients with Ischemic Cerebrovas-cular Disease at the Last Stage of Antiplatelet Agent / 中国药房

OBJECTIVE:To compare the incidence of cerebral events in patients with ischemic cerebrovascular disease compli-cating with cerebral microbleeds(CMBs)at the last stage of antiplatelet agent use. METHODS:140 patients with ischemic cerebro-vascular disease were selected from Beijing Anzhen Hospital Affilicated to Capital Medical University during Jan. 2013-Jan. 2014, and then divided into CMBs group and non-CMBs group according to whether complicated with CMBs,with 70 cases in each group. After followed up for 1 year(regular use of aspirin 100 mg/d and/or clopidogrel 75 mg/d),the incidence of recurrent cere-bral infarction and cerebral bleeding and mortality were compared. RESUTLS:The incidence of recurrent cerebral infarction was 12.9% in CMBs group and 8.6% in non-CMBs group,without statistical significance(P=0.412);the incidence of cerebral bleed-ing was 10.0% in CMBs group and 1.4% in non-CMBs group,with statistical significance(P=0.029);the mortality of cerebro-vascular event at the last stage was 5.7% in CMBs group and 4.3% in non-CMBs group,without statistical significance (P=0.698). CONCLUSIONS:The risk of cerebral bleeding increase in patients with ischemic cerebrovascular disease complicating with CMBs after the application of antiplatelet agent. For patients with ischemic cerebrovascular disease complicating with CMBs,the application of antiplatelet agent should be based on the complete judgment and weighing of benefit and bleeding risk.

Construction and effect of recombinant adenovirus vector containing siRNA targeting Raf-1 on cardiomyocyte hypertrophy in neonatal rats / 中国比较医学杂志

Objective To construct an adenovirus vector expressing small interfering RNA ( siRNA) targeting to rat Raf-1 gene and identify its function in cardiomyocytes.Methods The siRNA containing DNA sequence targeting to Raf-1 and its negative control sequence were designed, synthesized, annealed and subcloned into adenoviral shuttle vector pAdTrack-CMV.The recombinant adenovirus vector pAd-siRaf-1 was obtained by homologous recombination with pAdTrack-siRaf-1 linearized by PmeI and pAdeasy-1 in bacteria BJ5183, then transfected into HEK293 cells to package the adenovirus.Cardiomyocytes were infected with the adenovirus pAd-siRaf-1, and the expressions of Raf-1 and NF-κB protein were detected by Western blotting.[ 3 H ]-leu incorporation was evaluated by scintillation.The surface area of cardiomyocytes was measured using a HJ2000 image analysis system.Results The adenovirus vectors were verified by enzyme digestion and DNA sequencing.Compared with the Ang II group, Raf-1 and NF-κB expression, the surface area and [ 3 H]-leu incorporation of cardiomyocytes were significantly decreased in cardiomyocytes infected with the adenovirus PAd-siRaf-1.Conclusions A recombinant adenovirus vector containing rat siRaf-1 gene is successfully constructed.It can effectively reduce Raf-1 and NF-κB expression and cardiomyocyte hypertrophy induced by Ang II.

Development of a killed but metabolically active anthracis vaccine candidate strain / 生物工程学报

Anthrax is a zoonosis caused by Bacillus anthracis, which seriously affects human health. In recent years, a special phenomenon is found that the metabolic active of a bacterium remains after it is killed. To development of a KBMA (killed but metabolically active) Bacillus anthracis vaccine candidate strain, a plasmid pMAD and a recombinase system Cre-loxP were used to knockout the uvrAB gene of B. anthracis AP422 which lacks both of two plasmids pXO1 and pXO2. The results of PCR and RT-PCR shows that uvrAB genes were deleted from B. anthracis AP422 chromosome successfully. The constructed B. anthracis AP422deltauvrAB was inactivated by photochemical treatment (PCT) including an exposure in a long-wave-length ultraviolet (UVA) light and a treatment of 8-Methoxypsoralen (8-MOP), then the metabolic activity were detected by the method of MTS. The results showed that the killed B. anthracis AP422deltauvrAB maintained a highly metabolic activity for at least 4 hours, showing a state of KBMA. The KBMA strain of B. anthracis AP422deltauvrAB provides the prospective vaccine candidate strain for anthrax.

Antigen screening and identification of Helicobacter pylori outer membrane proteins from murine infection model / 中国免疫学杂志

Objective:To screen antigen of Helicobacter pylori outer membrane proteins by murine infection model.Methods:Parallel two-dimensional gel electrophoresis (2D) of outer membrane proteins extracted from Helicobacter pylori strain SS1 was performed.Western blot of a duplicate 2D gel hybridized with serum from H.pylori-infected murine was employed.Immunogenic H.pylori proteins identified in this way were digested in gel by trypsin and the mass of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).The data obtained from peptide mass finger-printing (PMF) were searched using the internet available database.Results:32 proteins were identified and they are in good agreement with typical protective antigens which reacted with serum from H.pylori-infected patients.Conclusion:The results suggest that murine model of H.pylori may be valid to screen antigens for human vaccination and the proteins identified in this paper are valuable for the selection of H.pylori protective antigens as well.

Expression of snake venom thrombin-like enzyme calobin in Pichia pastoris / 生物工程学报

Thrombin-like enzymes (TLEs) are studied widely because of their therapeutic potential in myocardial infarction and thrombotic diseases. We synthesized the DNA fragment encoding thrombin-like enzyme calobin from Agkistrodon caliginosus (Korean Viper) venom by fusion PCR and expressed it in Pichia pastoris. After induction by 0.5% methanol for 48 h, the expression level of recombinant calobin reached 3.5 g/L in medium. The recombinant calobin was purified by Q-Sepharose Fast Flow ion-exchange chromatography and Sephacryl-S-100 gel filtration chromatography. Purified sample had a molecular weight of 32 kD shown in SDS-PAGE. It hydrolyzed fibrinogen and formed a light white hydrolysis circle in fibrinogen plate. SDS-PAGE analysis showed that recombinant calobin cleaved Aalpha-chain of fibrinogen specifically, and produced an appropriately 40 kD new band. However, we failed to find its fibrin-clot formation activity.

Biomechanical analysis in the early period of flexor tendon healing after decimeter wave therapy / 中华物理医学与康复杂志

Objective To investigate the biomechanicai characteristics of flexor profundus tendons repaired after decimeter wave therapy, and to observe the effect of decimeter wave therapy on early active mobilization. Methods A total of 56 Leghorn chickens were randomly divided into a therapy group and a control group with 28 chickens in each. The 3rd and 4th toes of their left feet were employed for the establishment of a tendon injury model. The flexor profundus tendons were cut and repaired. Gypsum support was applied and fixed with an adhesive plaster after the operation. The operated sites on toes Ⅲ and Ⅳ were exposed. The external fixation was removed 3 weeks later and the chickens were left free to move. Decimeter wave therapy ( frequency 915 MHz, power 8 Watts) was ap-plied for 10 minutes once daily on the left foot of each chicken in the therapy group from day 1 until 3 weeks after the operation. Sham decimeter wave therapy was applied to chickens in the control group. Four chickens from each group were randomly selected at the 1st, 7th, 10th, 14th, 18th, 21st and 28th days for biomechanical analysis. Biome-chanical parameters including tensile strength of rupture (Pmax), elongation ratio at rupture (δimax) and the tensile adhesion strength of the rupture zone (W0>) were observed at each time point. Results At the 7th, 10th, 14th, 18th, 21st and 28th day after the operation, the differences in Pmax, δmax and W0 between the therapy and control groups were statistically significant. The results of the therapy group were better than those of the control group. Conclusions Local decimeter wave therapy after flexor tendon repair can promote intrinsic healing and reduce ex-trinsic healing. The speed and quality of healing are improved. The elasticity and tenacity of the injured tendons are enhanced. Therefore decimeter wave therapy is helpful for early active mobilization training.

Cost-Effectiveness Analysis of Three Therapeutic Schemes for Lower Respiratory Tract Infection / 中国药房

0.05);the cost-effectiveness ratios were 768.31,812.14 and 457.98,respectively.CONCLUSION:Scheme C was proved to be the best therapeutic regime.

Clinical Observation on Therapeutic Effect of Breviscapine Injection on Migraine / 中国药房

OBJECTIVE:To observe the therapeutic effect of Breviscapine injection in treating migraine. METHODS: 81 patients with migraine were randomly divided into two groups: 42 in treatment group were treated with Breviscapine injection 20 mL added to 5% Glucose injection 300 mL iv gtt q.d, and 39 in control group with Flunarizine 10 mg po quaque nocte. The treatment lasted for 2 courses (1 course of treatment lasted 14 days) with an interval of 15 days between the two courses. The patients were followed for more than 6 months. The curative effects between the 2 groups were compared, and the variations in hemorheological indexes of the 2 groups before and after treatment were observed. RESULTS:The efficacy in the treatment was signficantly higher than in the control gorup(P

Estimation of the height by measurement results of the body parts / 中国法医学杂志

In order to study the relationship between the height and the measured results of body parts,the length of the foot,the width of the foot palm,the whole length of the hand,and the length of the forfinger,middle finger,little finger as well as the height of 489 young people were measured.The data was then analyzed by the digital computer.The results showed that the linear relationship existed between the height and the measurement results of six body parts mentioned above in males.The linear regression equations were as follows:(1)y=93 55x+3 26x(by the length of the foot);(2)y=130 79+4 18x(by the width of the foot palm);(3)y=76 15+5 33x(by the whole length of the hand);(4)y=108 61+8 76x(by the length of the forefinger);(5)y=94 85+9 47x(by the length of middle finger);(6)y=120 79+8 68x(by the length of the little finger).Also,the linear relationship existed between the height and the whole length of the hand and the length of the middle finger,The regression equations were as follows:(1)y=119 86+2 44x(by the whole length of the hand);(2)y=129 89+4 08x(by the length of the middle finger).There is not any linear relationship existed between the height and the measurement results of these rwo body parts as mentioned above.