The feasibility of frontal chromatography for determining the complexation stability constant KML and total mole of binding site Λo was demonstrated by the accuracy and precision binding experiments between metal ions (Cu2+, Ni2+ and Co2+) and chelating ligand (IDA), in which R2>0.98 and RSD <5%.To further prove the universality of the frontal chromatography, the changing rules of KML values between Cu2+, Ni2+, Co2+ and IDA, Asp, L-Glu were examined under NaAc-HAc, Na-PB and Tris-HCl buffer systems.The results showed that the binding strength of chelating ligands for metal ions followed IDA>Asp>Glu;binding strength of metal ions for chelate ligands followed Cu2+>Ni2+>Co2+;and the binding effect with NaAc-HAc buffer was the best.In aqueous solution, quantum computing of binding energy (ΔE) and gibbs free energy (ΔG) between chelating ligand and metal ion was performed at the M06/6-311++G (d, p) level.According to ΔE and ΔG, the binding rules between chelating ligand and metal ion were predicted theoretically.These rules were basically in agreement with above experimental results.The present work provided effective method for studying on binding characteristics of metal ions for aminocarboxyl chelating ligands, thus exhibited a good foundation for improving the stability of immobilized metal affinity chromatographic column and solving the leakage of metal ions from the column in the process of competitive elution.

Objective To optimize the technology conditions of ultrasonic-enzyme-assisted extraction for sanguinarine from Zanthoxylum nitidum.Methods Extraction rate of sanguinarine determined by HPLC was served as an index.The applicability of the extraction solvent added with acid and enzymatic hydrolysis pretreatment to the ultrasonic-enzyme-assisted extraction of Zanthoxylum nitidum was investigated.Ultrasonic power,extraction frequency and solvent volume were optimized by orthogonal experiment.Finally,ultrasonic extraction time was optimized in dynamic process.Results The optimal process was as follows:Zanthoxylum nitidum powder was extracted 3 times by ultrasonic-wave (250 W) with 40％ ethanol (0.2％hydrochloride) as solvent (extracted for 15 rmin with 6-fold solvent at the first time,then extracted for 12 min with 3-fold solvent at the second and the third time,respectively).The extraction rate of sanguinarine was 88.6％.Conclusion The process is economic,efficient,energy-and time-saving,and provides experimental basis for industrial production of sanguinarine.

Objective To analyze the predicting value of Cornell-QRS standard and Sokolow-Lyon voltage to left ventricular hypertrophy in hypertensive patients.Methods One hundred and seventy-four patients with primary hypertension were enrolled, who were divided into left ventricular hypertrophy group (LVH group,n=50)and non-LVH group(n=124).The blood pressure, Cornell-QRS standard and Sokolow-Lyon voltage were collected and compared in the following-up period.Results Compared with non-LVH group,in LVH group the history of hypertrophy was longer(P <0.05),percent of grade 3 hypertrophy was higher(P <0.05),and 24 h SBP was higher(P <0.05).During the following-up of 6 months,1 year,2 year,the SBP (systolic blood pressure), DBP (diastolic blood pressure),Cornell-QRS standard and Sokolow-Lyon voltage in LVH patients were all significantly decreased (P <0.01).Compared with non-LVH group,the incidence of non-fatal myocardial infarction and stroke was higher in LVH group[ 3(6.0%)vs .1(0.8%),P <0.05;6 (12.0%)vs .2 (1.6%),P <0.01 ].By Cox analysis,Cornell-QRS standard and Sokolow-Lyon voltage were respectively independent predictors to non-fatal myocardial infarction and also stroke.Conclusion Cornell-QRS stand-ard and Sokolow-Lyon voltage may be independent predictors to main cardiovascular accident in hypertensive patients.

ObjectiveTo explore the relationship between the expression of smooth muscle 22 alpha ( SM22α),α-smooth muscle actin (α-SMA) and the development of varicose veins of the lower extremities. MethodsWe collected tissues of varicose saphenous vein in the experimental group,and what left over of normal saphenous vein tissues harvested for a coronary bypass operation ( control group).RT-PCR were applied to investigate the expression level of SM22αmRNA and immunohistochemical techniques were applied to investigate the expression level of SM22α and α-SMA protein in normal and varicose veins.ResultsThe expression level of SM22α mRNA in normal and abnormal veins was 0.2614 ±0.1168 and 0.5114 ± 0.1554 respectively ( t =5.020,P ＜ 0.01 ).The immunohistochemical staining showed that signals of SM22α and α-SMA protein existed in the cytoplasm of smooth muscle cells.The integrate optical density (IOD) of SM22α in normal and abnormal veins was 10 055 ± 3584 and 16 226 ± 3378 respectively ( t =4.991,P ＜ 0.01 ).The IOD of α-SMA in normal and abnormal veins was 9746 ± 3903 and 15 371 ± 4318 respectively (t =3.861,P ＜ 0.05 ). ConclusionsSM22α and α-SMA was down-regulated in varicose veins,indicating that the VSMC had transformed the typical contractile phenotype into the typical synthetic phenotype.Phenotype changes of VSMC further promote venous wall remodeling leading to the occurrence of varicose veins.

Objective To investigate the clinical efficacy of multi-functional fire needling plus auricular plaster therapy for moderate to severe acne vulgaris.Methods One hundred and fifty patients with moderate to severe acne vulgaris were randomly allocated to treatment and control groups, 75 cases each. Both groups were first given bloodletting at Ear apex and Helix 4. Then the treatment group received multi-functional fire needling plus auricular plaster therapy and the control group, auricular plaster therapy alone. The symptom and sign score was recorded in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups.Results There was a statistically significant pre-/post-treatment difference in the symptom and sign score in the two groups (P<0.01). There was a statistically significant post-treatment difference in the symptom and sign score between the two groups (P<0.05). The total efficacy rate and the cure and marked efficacy rate were 98.6% and 88.7%, respectively, in the treatment group and 93.2% and 60.8%, respectively, in the control group. There was a statistically significant difference in the cure and marked efficacy rate between the two groups (P<0.05).Conclusion Multi-functional fire needling plus auricular plaster therapy is an effective way to treat moderate to severe acne vulgaris.

Objective To determine the effect of different serum insulin-like growth factor-Ⅰ(IGF-1)levels on mouse cancer.Methods A total of 120 male mice at 6 weeks of age(60 control mice and 60 LID mice)were subcutaneously injected colon tumor CT26 cell line.Each group was random divided into two subgroups respectively,every 10 mice of one subgroup were injected subcutaneously with growth hormone(GH)(1ms/kg)daily from the 10th,14th and18th days respectively until the 22nd days,and the other subgroup received saline injection.Results All mice treated with GH have higher level of IGF-1,compared with those treated with saline.High level of IGF-1 promoted the development of cachexia in these mice treated with GH from the 10th days.However,the level of IGF-1 has negative correlation with the cancer cachexia state for mice treated with GH from the 14th days.Conclusion Circulating IGF-1 and GH play an important role in tumor growth 4nd cachexia development in the early stage of cancer and can ameliorate the state of cachexia in the advanced stage.

Objective To study the effect of a new mucA gene mutation on the biofilm formation process and the morphology of matured biofilm of Pseudomonas aeruginosa. Methods The mucA gene of PAO1 was cloned into the Pseudomonas aeruginosa expression plasmid pUCP20. The recombinant plasmid was transformed to mucoid PA17 which contained a new mucA mutation. Positive clones of the plasmids were identified by enzyme digestion and sequencing. The expression levels of algD in the positive clones were assessed by semi-quantitative RT-PCR. The modified plate culture method was used to establish the biofilm models of PA17, PA17 with recombinant plasmid and PAO1 in vitro. Results Transformation was identified by the decreased expression of algD in positive clones. The rate of biofilm formation of the positive clones was between those of PAO1 and PA17. The irreversible adhesion occurred after 8 h and the matured biofilm was observed on day 6. The morphologies of PA17, PAO1 and PA17 with recombinant plasmid were the same. Conclusion The mucA gene mutation of PA17 delays the formation of irreversible adhesion of PA17 biofilm, but it has no effect on the morphology of matured biofilm.

Objective Toinvestigatetheriskoflong-termrecurrenceofstrokeanditspredictorsin youngpatientswithischemicstroke/transientischemicattack(TIA).Methods Theclinicaldataofthe consecutive young patients (18-45 years)with ischemic stroke/TIA (within 2 weeks after onset)admitted to the department of neurology in the First Affiliated Hospital of Sun Yat-Sen University between August 2008 and July 2013 were enrolled prospectively. All patients were regularly followed up for a long time (The patients were followed up at the 1 st,6 th,and 12 th month after onset;then they were followed up once for every 6 months)in order to investigate stroke recurrence. The Kaplan-Meier curves were used to analyze the cumulative stroke recurrence rate of all patients. The last contact time for patients lost to follow was used as censored data to be enrolled in the analysis. The univariate analysis of the related risk factors for stroke recurrence using Log-rank test. Multivariate Cox proportional hazard regression was used to detect the related risk factors associated with stroke recurrence (adjusting for age and sex). The variables of the results of Log-ranktestP≤0.1wasselectedandenrolledinthemultivariateregressionanalysis.Results Atotalof 312 patients were enrolled in the analysis,including 294 with ischemic stroke and 18 with TIA. Their mean follow-up time was 34 ± 19 months. Thirty-four patients had recurrent stroke,including 23 with ischemic stroke,7 with TIA,and 4 with cerebral hemorrhage. The cumulative recurrence rates of stroke at 1 ,3 , and 6 years after onset were 6. 2%,10. 3%,and 16. 4%,respectively. The results of multivariate Cox proportional hazards regression analysis showed that hypertension (risk ratio [RR]2. 159;95% confidence interval [CI]1. 048-4. 447,P=0. 037)and cardioembolism (RR,2. 869;95%CI 1. 119-7. 357,P=0.028)weretheindependentpredictorsforstrokerecurrence.Conclusion Theoverall6-yearriskof recurrent stroke is not high in the Chinese young patients with ischemic stroke/TIA,but the risk of stroke recurrence is relatively higher in the first year. Hypertension and cardioembolism are the potential predictors of stroke recurrence;therefore,attention should be paid in clinical practice.

The holding of the national college students' clinical skills competition reflects the importance of medical education for clinical practice training . Although through intensive itemized skills drills, the competitors can complete each individual operation with satisfaction, due to the lack of clinical experience, in the integrated circuit training, they will easily stray into question stemtrap. The concrete analysis of the national college students' clinical skills contest competition reflects the medical students' insufficient recognition of clinical skills, lack of the overall concept of the intensive medical treatment, not flexible and insufficient mastery of the connotation of the clinical skills, which seriously restricts the improvement of clinical education. Therefore this article raises special sugges-tions, referring to training focusing on “Airway and Circulation”, developing the critical care thinking based on the relationship of multi-organ and improving the first-aid capability of the team work, so as to provide reference for the improvement of training effect.

Objective To study the relationship between plasma free fatty acids composition and the incidence of nonalcoholic fatty liver disease(NAFLD) .Methods By the design of case‐control study ,105 patients with NAFLD as cases and 110 healthy peo‐ple as controls were enrolled into the study .Plasma free fatty acid levels were determined by gas chromatography .Results High level of plasma palmitic acid(C16 :0)(OR=1 .769) was the risk factors of NAFLD ,while plasma levels of linoleic acid(C18 :2 n‐6) (OR=0 .855) and arachidonic acid(C20 :4 n‐6)(OR=0 .181)were negatively associated with the incidence of NAFLD .Conclusion These findings suggest that a proper ratio of diet fatty acids intake may reduce the risk of NAFLD .