Objective To study on the drug-resistance mechanism of Brucella resistance to Quinolone antibiotics to guide the selection and use of antimicrobial agents in clinical practice. Methods Six strains of Brucella melitensis(Bru1, Bru2, Bru3, Bru4, Bru5, Bru6) were selected to be induced resistance to levofloxacin in vitro respectively. The MICs of the 6 strains of Brucella melitensis and induced resistant strains were measured by agar dilution method. The sensitivity to Quinolone antibiotics (Levofloxacin, Ciprofloxacin, Lomefloxacin, Norfloxacin, Fleroxacin, Ofloxaein) of 6 strains of Brucella melitensis and induced resistant strains was measured by K-B method. The gyrA of the 6 strains of Brucella melitensis and induced resistant strains was amplified by PCR, then the nucleotide sequence of the genes were analyzed. Results The MICs of Bru1,Bru2,Bru3,Bru4, Bru6 were 0.50 μg/ml and Bru5 was 0.25 μg/ml. The strains Bin3, Bru4 were induced into drug-resistant strains by Levofloxacin, then were named LEVR3 and LEVR4 respectively. The MICs of LEVR3 and LEVR4 were 64,128 μg/ml with 128 and 256 times higher than that of the parental strains. The 6 strains of Brucella melitensis were sensitive to Quinolone antibiotics, LEVR3 and LEVR4 were resistant to Quinolone antibiotics. Neucleotide sequence analysis and comparison of the derived amino acid sequence revealed that Quinolone resistance-determing region of GyrA had a substitution at position Ala87 and Asp91 in laboratory resistant strains. Conclusion The results of in vitro experiments show that acquired resistance of Brucella melitensis strains to Levofloxacin could beinduced when exposed to high level of some antibacterial agents for short term. Two drug-resistant strains occur mutations in gyrA and have cross-resistance to other Quinolones.

Objective To establish an HPLC method for the simultaneous determination of contents of gallic acid, corilagin and ellagic acid in aqueous extract from Sanguo Decoction. Methods The analysis was carried out on an Agilent C18 column (4.6 mm × 250 mm, 5 μm), and the mobile phase was composed of acetonitrile and 0.1%phosphoric acid aqueous with gradient elution. The detection wavelength was 260 nm. The flow rate was 1.0 mL/min at column temperature of 30 ℃. Results Gallic acid, corilagin and ellagic acid were completely separated and the peak shape was good. It showed a good linearity in the concentration range of 0.174–2.61 μg, 0.04–0.60 μg, and 0.052–0.78 μg, respectively. The average recoveries of gallic acid, corilagin and ellagic acid were 99.98%, 99.81%and 100.12%, respectively. The RSD were less than 2.0%. Conclusion This method is rapid, accurate and repeatable, which can provide references for the quality control of Sanguo Decoction.

Expression and regulation of the P-glycoprotein (P-gp) is associated with the development of multidrug resistance in cancer.Cancer cells are exposed in endogenous damages from its own micro-environments and inflicted stimulated damages.These harmful micro-environments could trigger cellular stress response and regulated the expression of P-gp through various activated signal transduction pathways.Researches of the cellular endogenous micro-environments and exogenous stimulation damages on the expression and regulation of P-gp would have important significance for exploring the multidrug resistance of cancer cell.

Objective The current status of achievement motive levels and autonomous learning ability of nursing students was studied and compared in order to put forward suggestions to improve the achievement motive levels accordingly.Methods 119 nursing students were investigated about their achievement motive levels and autonomous learning ability by questionnaires.Then these students were divided into three groups according to their achievement motive scores and the autonomous learning ability was compared.Results The scores of Success-oriented motive of nursing students were significantly higher than that of failure-threatened motive.The total score of autonomous learning ability was(88.653+9.373)and the autonomous learning ability among these three groups was statistically different(P＜0.01).Conclusion The achievement motive greatly influenced the autonomous learning ability of nursing students.Measures should be adopted to improve the achievement motives so as to enhance their autonomous learning ability.

Objective To evaluate the correlation between serum procalcitonin concentration and systemic inflam-matory response syndrome (SIRS)score in patients with bacterial bloodstream infection.Methods In January-De-cember,2012,96 patients with bacterial bloodstream infection in a hospital were selected as trial group,and these patients were divided into three groups(group A,B and C)according to SIRS score;84 patients without bacterial in-fection was as control group,PCT concentration of all patients were detected,and the correlation between PCT con-centration and SIRS score was analyzed.Results Among 96 patients with bacterial bloodstream infection,7 (7.29%)died (4 were in group B and 3 in group C);there was no death case in control group.PCT concentration in control group,group A,B and C of trial group were (0.28±0.09)ng/mL,(0.63±0.13)ng/mL,(3.68±1.01)ng/mL,and(7.45±1.53)ng/mL,respectively,the difference between each group was significant(P<0.01).Pairwise comparison of four groups showed statistical difference (P<0.001).Spearman correlation analysis on PCT concen-tration and SIRS score was conducted,correlation coefficient r=0.874(P<0.001)suggested positive correlation be-tween serum PCT concentration and SIRS score.Conclusion PCT concentration in patients with bacterial blood-stream infection and SIRS score is positively correlated,PCT concentration and SIRS score can be used as two mark-ers for assessing the extent and prognosis of bacterial bloodstream infection.

Objective To study the distribution and expression of Semaphorins-3A protein in brain of postnatal rats.Methods Semaphorins-3A positive cells were observed by immunohistochemistry in the cerebral cortex,hippocampus,dentate gyms and entorhinal cortex in postnatal 0d,7d,14d,21 d and 28d of Sprague Dawley rats.Results Semaphorins-3A positive cells widely distributed in the granule cell layer ( Ⅱ ),external pyramidal cell layer ( Ⅲ ),internal granular cell layer ( Ⅳ ),pyramidal cell layer ( Ⅴ ) and layer of polymorphous cells ( Ⅵ ),in addition to the molecular layer ( Ⅰ ) of the parietal,occipital,frontal,temporal,insular,cingulate cortex,piriform cortex and entorhinal cortex with postnatal 0d,7d,14d,21d and 28d rats.The amount of semaphorins-3A positive cells(IOD) in the entorhinal cortex was 84916.23 ± 3266.34 in P0d,77711.41 ± 2634.26 in P7d,74124.25 ± 3989.09 in P14d,65887.63 ± 3406.57 in P21d and 57705.96 ± 3136.35 in P28d,meanwhile the region of semaphorins-3A positive cells narrowed in the part level with Ⅱ -Ⅵ levels of cortex.Similarly semaphorins-3A positive cells distributed mainly in granule cell layer of dentate gyrus,CA1,CA3 region and only a few of semaphorins-3A positive cells scattered in the multi-line layer in hippocampus.The expression level of semaphorins-3Awas significant difference among postnatal 0d,7d,14d,21d and 28d rats (P＜0.01).Conclusion Semaphorins-3A positive cells widely distribute in the various cortex and hippocampus in developing rat brain,and the region of semaphorins-3A is reduced with age growth of rats.

ObjectiveTo observe Semaphorins-3A and synaptophysin P38 expression in hippocampus of developing rat induced by status epilepticus.Methods 320 SD rats were divided into four groups (P7,P14,P21 and P28) according to day -old after birth (7d,14d,21d and 28d).Rats in each group were randomly divided into model group (SE group) and saline control group (NS group).SE was induced by Pentylenetetrazol (PTZ).Semaphorins-3A and synaptophysin P38 expression were determined by immunohistochemical staining on 1d,7d,14d,21d and 28d after SE in hippocampal CA1 of rats.ResultsSemaphorins-3A-positive cells could be seen in the hippocampal granule cell layer in all rats.Semaphorins-3A expression tended to decrease with the increasing of day-age,especially in P7 group(91 552.68 ± 4664.69 ).No matter how day-age,Semaphorins-3A expression was similar to that in NS group and was obvious reduced in 7d after SE(56 938.84 ± 5688.47 ).Meanwhile P38 expression in P7-day-age rats had had been gradually increasing between 1 day and 14d (5413.18 ±48.77,6223.40±29.19,6902.94 ±78.51 ) and then stabilized gradually on 21d(7523.42 ± 62.94) after rats were tested.P38 expression in other day-age rat was relatively stable on the same level in physiological state.On the other hand P38 expression in the hippocampal CA1 region of P7,P14,P21 and P28 rats was significantly higher than that in normal rats between 1day and 28day after SE episode(P＜ 0.05 ),and reached a peak on 14 day(8408.35 ± 55.73 ).ConclusionSemaphorins-3A and synaptophysin P38 involved in hippocampal synaptic plasticity of rat in developing stage and epilepsy.

Objective To observe neural stem cells proliferation,migration and differentiation in hippocampus in developing rats with status epileptictus.Methods 320 healthy SD rats at age 7,14,21,28 d (P7,P14,P21,P28) were randomly divided into status epilepticus (SE) and normal control group.In each group those rats at the same age were further randomly divided into 1,7,14,21,28 d five time points after PTZ-induced SE (n =8).New cell proliferation and migration were observed by immunohistochemistry studies in the dentate gyms.Double labeling with Brdu/NeuN and Brdu/GFAP was performed in the P14 rats.Results Nestin positive cells appeared in the dentate gyms on 1 d after SE in P7,P14,P21,P28 rats.The number of nestin positive cells gradually increased on 7 d and reached a peak on 14 d,then gradually reduced on 21 d,finally fell to a minimum on 28 d after SE.The numbers of nestin positive cells on 7 d(177.00 ± 3.22,t =16.033) and 14 d (195.00 ± 3.41,t =28.840) were significantly higher in the SE group than the NS group (147.50 ± 2.08,136.50 ± 2.65,both P ＜ 0.05).The smaller age of rats with SE onset,the greater the nestin intensity.But the number of nestin positive cells in the dentate gyrus of normal rats were gradually decreased with increasing age.Nestin positive cells were distributed in subgranular zone of dentate gyrus on 1 d and 7 d after SE,then gradually migrated to the granule cell layer on 14 d with morphological changes.Small part of nestin positive cells were ectopically migrated to the hilus of dentate gyrus in P14,P21,P28 age rats,and were also seen in the CA1,CA3 of hippocampus and cortex with various cell morphology.For differentiation of newly generated cells,most of Brdu positive cells coexpressed NeuN and about 4％-5％ cells co-expressed GFAP.Conclusions SE could induce neurogenesis in the hippocampal dentate gyrus area in developing rats which has age-related characteristics.Most new cells migrate from the subgranular zone to the granule cell layer of the dentate gyrus,and a small number of newly generated cells ectopically migrated to the hilus of dentate.The majority of newly generated cells differentiate into neurons,and the others differentiate into glial.

A new risk analysis method based upon reliability theory was introduced in this paper for the quality risk management of Chinese medicine injection manufacturing plants. The risk events including both cause and effect ones were derived in the framework as nodes with a Bayesian network analysis approach. It thus transforms the risk analysis results from failure mode and effect analysis (FMEA) into a Bayesian network platform. With its structure and parameters determined, the network can be used to evaluate the system reliability quantitatively with probabilistic analytical appraoches. Using network analysis tools such as GeNie and AgenaRisk, we are able to find the nodes that are most critical to influence the system reliability. The importance of each node to the system can be quantitatively evaluated by calculating the effect of the node on the overall risk, and minimization plan can be determined accordingly to reduce their influences and improve the system reliability. Using the Shengmai injection manufacturing plant of SZYY Ltd as a user case, we analyzed the quality risk with both static FMEA analysis and dynamic Bayesian Network analysis. The potential risk factors for the quality of Shengmai injection manufacturing were identified with the network analysis platform. Quality assurance actions were further defined to reduce the risk and improve the product quality.
Drugs, Chinese Herbal ; administration & dosage ; chemistry ; standards ; Medicine, Chinese Traditional ; standards ; Quality Control ; Reproducibility of Results ; Risk Assessment

AIM: To establish the method of determining psoralen and isopsoralen in Bushen Yinao Capsules(Fructus Psoraleae,Radix et Rhizoma Ginseng Rubra,Poria,etc.). METHODS: HPLC conditions were as(follows:) Chromasil C_(18) column,a mobile phase of CH_3OH-H_2O(55∶45),and the wavelength at 247 nm. RESULTS: This method showed a good linear relationship in the ranges of 0.038-0.309 ?g(psoralen) and 0.036-0.295 ?g(isopsoralen).The average recoveries were 99.50% for psoralen(RSD was 0.91%,n=5) and 100.52% for isopsoralen(RSD was 1.70%,n=5). CONCLUSION: The method is simple,swift,accurate and practical in quality control of Bushen Yinao Capsules.