OBJECTIVE:To establish a HPLC-chiral stationary phase(CSP) for separating the enantiomers of lactic acid and determining the optical purity of lactic acid.METHODS:Lactic acid samples of different optical purity were determined by HPLC-CSP vs.optical rotation method.The enantiomers of lactic acid were separated on Chirex 3126(D)-penicillamine chiral column.The mobile phase consisted of copper sulfate solution-isopropyl alcohol (85∶15) at a flow rate of 0.6 mL?min-1.The detection wavelength was set at 254 nm.RESULTS:A baseline separation of the enantiomers of lactic acid was achieved with a separation factor of greater than 1.2.The calibration curve was linear in the range of 0.75～48.24 mg?mL-1 (r=0.999 3),RSD=1.21%.The determination results determined by HPLC-CSP were similar to those determined by optical rotation method.CONCLUSION:The established method is simple,reliable in results and reproducible,and it is applicable for the separation of the enantiomers of lactic acid and determination of the optical purity of lactic acid.

Objective To evaluate the long-term effective rate,retention rate and tolerability of the ketogenic diet (KD) in pediatric drug-resistant epilepsies.Methods Data of 36 children who were treated in Department of Pediatrics,the Second Affiliated Hospital of Xi'an Jiaotong University from Nov.2011 to Dec.2013 and had continuous follow-up of at least 12 months after initiation of the KD were analyzed prospectively.Response was defined as 50％ seizure reduction.The effective rate,retention rate, outcome-predictive value of various clinical factors were also assessed.The causes of the patients withdrew from KD and side effects were recorded and analyzed.Results Thirty-six children(29 boys,7 girls; mean age of 2.84 years)were included.The effective rate was 50.0％,52.8％,47.2％ and 41.7％ at the 1,3,6,12 months;the retention rate respectively was 94.4％,91.2％,69.4％ and 52.8％.Seventeen cases withdrew from KD.Seven cases (41.2％)ceased KD becau~ of a lack of compliance,5 patients because of poor outcome,and 2 cases because of repeated infection.None of the age,disease duration,etiology and seizure type could be used as the predictor for the favourable treatment for outcome.The antiepileptic drugs before KD might be predicted the efficacy of the KD.The less amount of the antiepileptic drugs,the more opportunity of the KD might have to succeed.The side effects on the start-up period were drowsiness,week and digestive discomfort,hyperlipoidemia,hypoglycemia and hepatic dysfunction.The side effects on the maintenance period were digestive discomfort,susceptibility to infection,hyperlipoidemia,the deficiency of trace elements.Conclusions The KD is a safe and potentially effective method in treatment of refractory epilepsy patients who do not respond to customary medication therapies.

Objective To study the effect of cyclooxygenase-2 (COX-2)selective inhibitor Celecoxib on the expression of P-glycoprotein(P-gp)in the brain of rats with status epilepficus,in order to assess the therapeutic value of intractable epilepsy.Methods Sixty adult male SD rats were randomly divided into blank control group,the epilepsy model group and Celecoxib intervention group.The status epilepticus was induced in rats by injecting Lithium pilocarpine.Forty-eight rats were included in the experiment.There were 16 rats in each of the blank control group,epilepsy model group and Celecoxib intervention group,respectively.Immunohistochemical method and Western blot method were used to detect the expression of P-gp in experimental group in the frontal cortex and hippocampus.Results Immunohistochemistry result showed that the expression of P-gp was significantly higher in epilepsy model group than the blank control group,and the difference was statistically significant (P ＜ 0.01);The P-gp expression in the Celecoxib intervention group was lower than that in the epilepsy model group,and the difference was statistically significant (P ＜0.01).Western blot results suggested that the expression of P-gp could be found both in the frontal cortex and hippocampus in each group.Compared with the blank control group,the P-gp expression was significantly higher than that in the epilepsy model group,and the expression of the P-gp was lower after the Celecoxib intervention than that in the epilepsy model group,and the difference was statistically significant (P ＜ 0.05).Conclusions COX-2 inhibitor Celecoxib could decrease the expression of P-gp in brain tissue with status epilepticus,which may provide a new method for the treatment of intractable epilepsy.

Objective To understand teachers' cognition and behavior of PBL teaching in nursing college.Methods Eighty teachers were investigated by questionnaire at eight nursing colleges in Hunan province.The percentage was used to do statistical description.Results Totally 75 copies of valid questionnaires were returned,the effective recovery rate was 93.75％.Among all,96％ of teachers thought that teacher was the guide;85.3％ teachers thought that students were knowledge's seekers;93.3％ teachers heard of PBL teaching;81.3％ teaches thought that PBL teaching played a positive role ;74.7％ teaches understood PBL teaching by communication or reading literature; 72％ teachers were willing to try PBL teaching;46.8％ teachers emphasized on multidisciplinary knowledge fusion in teaching; 88.3％ teachers encouraged student discussion in class.The main factors effecting PBL teaching were case and problem design,teaching result evaluation,lack of expert guidance and teacher instruction.Conclusion Teachers have certain understanding of PBL.Organizers should formulate related policies for teachers and students in order to create good education environment.

An evaluation system including experiment preparation,experiment process,com-prehensive design of experiment,experiment skills and written exam was established in order to adapt to the experimental teaching reform in our school. Experimental preparation assessment was to evaluate‘preview and self-evaluation report’prepared and submitted by students. Experimental process assess-ment was to evaluate students' classroom performance and experiment reports. Assessment of compre-hensive design experiments was to evaluate the overall participation of students. Skill assessment was consisted of oral test and experiment operation test. Final written examination,mainly consisting of subjective questions,emphasized on student's flexible use of knowledge and ability to solve practical problems. The evaluation system of promoting student's learning and teacher's teaching through the examination not only fully arouse student's attention on experiment,but also make teachers more ob-jectively and really understand students' learning situation and the teaching effectiveness.

Objective To investigate the rule of the cerebral tissues aquaporin-4(AQP-4) expression in acute and chronic hepatic failure mice.To study the molecular biologic mechanism of the diffusion weighted imaging(DWI).Methods Sixty five male Sprague-Dawley rats were divided into 3 groups randomly,including acute(n=25),chronic hepatic failure(n=25)and control group(n=15). Thioacetamide(TAA)intraperitoneal injection produces the acute and chronic hepatic failure models.All rats in groups were examined with MR DWI.We Observed the distribution of abnormal signal on DWI.The DWI single values of top and lateral cortex of parietal lobe,peripheral region of lateral ventricle in the highest hyperintensity section of brain were measured.Blood ammonia values were examined.The pathologic and immuno-histochemistry and RT-PCR examination for brain specimen were performed.All date were analyzed with statistical methods.Results The mean values of blood ammonia were significantly different (P0.05).Conclusions Increase of the blood ammonia was the main cause for the brain energy metabolic abnormality and AQP-4 mRNA and protein expression.The hyperammonemia was the key factor in the occurrence and development of the hepatic brain edema.The abnormal findings in DWI signal could reflect the range and degree of the brain edema and AQP-4 protein expression.

Objective To explore the relationship between blood lipids level with creatinine clearance rate(Ccr)in patients with heart failure(HF). Methods A total of 955 patients who were diagnosed with heart failure(cardiac function NYHAⅡ~Ⅳclassification)upon discharge from the Department of Cardiology of the Second Hospital of Tianjin Medical University, between January 2010 to June 2013 were enrolled as HF group. Healthy adults (n=200) with normal cardiac function which approximately matched basic condition with HF group were selected as control group. The HF group was fur?ther divided intoⅡ,Ⅲ,Ⅳclassification according to their cardiac function(NYHA classification). HF group was also divid?ed into normal renal function group, mild renal injury group and moderate-severe renal injury group based on their Ccr. Ef?fect of gender and lipid parameters were also compared. Binary logistic regression was used to analyze factors influencing renal function in patients with HF. Results Compared with people in the control group, the levels of triacylglycerol(TG), total cholesterol(TC), low density lipoprotein cholesterol(LDL-c)and non high density lipoprotein cholesterol(non-HDL-c)in patients of HF group were increased while Ccr and high density lipoprotein cholesterol(HDL-c)were decreased. Ccr and lipids were obviously decreased in patients with HF of Ⅳclassification. TG and HDL-c were decreased in moderate-severe renal injury group. Females had a higher lipid levels than males in HF group(P<0.05 or P<0.01). Advanced age, coronary heart disease and hypertension were all risk factors for renal impairment in patients with HF by binary logistic re?gression. On the other hand, weight gain and HDL-c were the protection factors for renal function in HF patients. Conclu?sion Dyslipidemias may lead to renal insufficiency in patients with HF. It was important to control lipids and improve re?nal function in patients with HF.

Objective To identify the specific genotype and analyze clinical features of spinocerebellar ataxias (SCAs) pedigree in the region of Yunnan Province.Methods Fourteen SCAs pedigrees and 183 blood samples of the family members were collected between January 2011 and July 2014 from Department of Neurology,First Affiliated Hospital of Kunming Medical College.Polymerase chain reaction (PCR) amplification,agarose gel electrophoresis and DNA sequencing technologies were utilized to identify the specific genotype of SCAs pedigree.Presymptomatic tests were carried out and the clinical features and genetic test results of patients were carefully analyzed.Results SCA3 was the most common subtype of SCAs in the Han nationality of Yunnan region.Nine of the 14 families were SCA3,only one family was SCA2.Additionally,there were four SCAs families that remained indeterminate.The patients with di-allele mutations (46/77) of SCA3 gene had early onset,rapid progression and serious clinical symptoms.Hereditary SCA3 and autonomic dominant polycystic kidney disease can happen simultaneously in a family.The proband SCA3 gene' s CAG repeat number is 28/76,and repetitions of the mutation allele are in all range.The PKD1 gene exon 23 is found to be in abnormal sequence.Conclusions SCA3 is the most common subtype of SCAs in the Han population of Yunnan region.There are 15/46 incomplete penetrance nutation and 46/77 di-allele mutations.It is possible that di-allele mutations make the disease worse and accelerate clinical course progression.SCA3 and polycystic kidney disease can uncommonly happen simultaneously in a family,which perhaps suggests there are interactions between the two disease-virulence genes.

Objective To explore the effect of nursing of Traditional Chinese Medicine on cognitive function among aged people with mild cognitive impairment in elderly care institution. Methods A total of 66 cases with mild cognitive impairment elderly that previously were screened out from two pension institutions in Taiyuan city as the research objects. According to the endowment institutions where the objects of study dwelled was divided into control group (34 cases) and intervention group (32 cases). The intervention group accepted nursing of Traditional Chinese Medicine plan for four months. The control group received the disease related knowledge health education. The cognitive function of the two groups were assessed using the Montreal Cognitive Scale (MoCA) before and after the intervention. Results Before the intervention, two groups had no statistical significance in each dimension score and total score of MoCA. After the intervention, four dimensions scores including ability of visual spatial and execute , attention , delayed recall , directional force and the total score of the intervention group respectively were (2.67±1.09) points, (4.07±1.08) points, (1.97±0.81) points, (4.83±0.99) points, (18.80±1.74) points while the control group were (2.15 ± 0.71) points, (3.30 ± 0.63) points, (1.39 ± 0.97) points, (4.24 ± 1.41) points, (15.06 ± 2.98) points, and the difference was statistically significant (t=1.91-6.06, P < 0.05 or 0.01) ;Before intervention the language score, delayed recall score and the total score of the intervention group were (1.43±0.77) points, (1.57±1.10) points, (17.27±1.99) points, respectively while after the intervention were (1.80 ± 0.66) points, (1.97 ± 0.81) points, (18.80 ± 1.74) points, there was statistically significant difference (t =-2.16,-2.11,- 4.34, P < 0.05 or 0.01). Conclusions Nursing of Traditional Chinese Medicine can delay the process of cognitive decline of mild cognitive impairment in the elderly, and have a certain effect on prevention and treatment of patients with mild cognitive impairment.

Objective To investigate the clinical effect of large dosage of Methylprednisolone on epilespy combined with electrical status epilepticus during sleep(ESES) in children.Methods Forty-six epielpsy patients with ESES were treated with additive large dosage of Methylprednisolone.The seizures and video electroencephalogram discharges were observed before and after using Methylprednisolone.The Methylprednisolone effect in eliminating the methylprednisolone of ESES and controlling of clinical seizures and improving cognitive function were analyzed.Results Two cases were lost and 44 cases were included,male 30,female 14,the age at onset was (5.37 ±2.52) (2-9) years old.The duration of follow-up was (4.12 ± 2.00)(1-9) years old.The age at diagnosis of ESES was (8.17 ± 2.09) (4.0-12.5) years old.The efficacy of Methylprednisolone on seizures was 72.7％ (32/44 cases),while the efficacy of Methyl prednisolone on electroencephalograph (EEG) was 59.1％ (26/44 cases).For patients who were resistant to Levetiracetam or Clonazepam,Methylprednisolone was still effective.Intelligence quotient had no significant changes before and after treatment(P ＞ 0.05).The earlier onset age,the worse effect of Methylprednisolone.The efficacy of methylprednisolone for atypical benign epilepsy with cento-temporal spike(BECT) was higher than other syndromes.Conclusions Large dosage of Methyl prednisolone therapy for children with ESES,especially for those resistant to traditional or new antiepileptic drug for ESES,was effective and safe.The onset age and syndrome classification may have a certain value for prognosis and prediction of the effect of Methylprednisolone treatment.