Object To study the chemical constituents in the ro ot of Isatis indigotica Fort Methods The s eparations were carried out by column chromatography and identified by advanced physical and spectral data analysis Results Five compounds were isolated and identified as neohesperidin (Ⅰ), ammonium formate (Ⅱ), isol iquiritigenin (Ⅲ), liquiritigenin (Ⅳ), and adenosine (Ⅴ) Conclusio n Neohesperidin, liquiritigenin and isoliquiritigenin are isolated f o r the first time from the plants of Cruciferae, ammonium formate is obtained fro m the root of I indigotica for the first time

Rabies is a deadly zoonos caused by rabies virus, the case fatality almost is 100%following suffering from this dis-ease.Recently, the incidence of rabies tends to increase in China.A rapid, accurate, economical, simple and convenient, easy to be widely used method for rabies virus detection in laboratory is urgently needed.This paper reviews the nucleic acids diagnosis techniques of rabies virus from the nucleic acids detection methods based on PCR, NASBA, LAMP, Genechips and analyzes the advantages and disadvantagesof the above-mentioned methods, which provides a reference for rapid laboratory diagnosis of rabies virus to efficently prevent and control rabies.

Objective:To observe the influence of long-term exercise training on carotid intima-media thickness (IMT)in patients with mild-to-moderate hypertension.Methods:A total of 92 patients with mild-to-moderate es-sential hypertension (EH)were randomly and equally divided into exercise group and routine treatment group.Exer-cise group received exercise training based on routine treatment.All patients were followed up one year.Influence of long-term exercise training on carotid artery diameter and carotid IMT was observed in EH patients.Results:Af-ter one year,there were significant reductions in systolic blood pressure (SBP),diastolic blood pressure (DBP),ca-rotid artery diameter and carotid IMT in both groups,P<0.05 all;compared with routine treatment group,there were significant reductions in SBP [(145.72±11.31)mmHg vs.(130.89±13.01)mmHg],DBP [(88.49±7.32) mmHg vs.(81.71±8.45)mmHg],carotid artery diameter [(6.34±1.23)mm vs.(6.22±1.01)mm]and carotid IMT [(0.89±0.21)mm vs.(0.84±0.11)mm]in exercise group after treatment,P<0.05 all.Conclusion:Long-term exercise training can effectively reverse early arteriosclerosis lesion of carotid wall and effectively control blood pressure.

Objective By means of value-added reflection of students' learning behavior, this article gives an objective description of the current situation of pharmaceutical undergraduate education and puts forward some suggestions to strengthen the connotation construction of pharmaceutical education.Method Using the Chinese College Student Survey (CCSS) developed by Tsinghua University and Indiana University (US), 1100 students participated in a one-to-one web-based anonymous questionnaire, and using SPSS19.0 to analyze the data.Five comparable indexes and students' recognition behaviors were discussed using descriptive analysis and independent sample t test.Result The five comparable index scores and learning behavior skill index scores of the students in Chinese traditional medicine specialty and pharmacy were higher than the national overall level of all kinds of colleges and universities of science categories.Compared with the national norm of medical colleges, except that the score of 43.36 of the five comparable indexes in pharmaceutical professional academic challenge (level of academic challenge, LAC) was less than 43.63 national medical colleges LAC norm level, the other index scores were higher.LAC and active collaborative learning scores of Chinese pharmaceutical majors were lower than those of pharmacy students, and the difference was statistically significant (P＜0.05).In the study of behavioral skills, the scores of two major students' use, evaluation and comprehensive behavior were lower than the national standard of medical institutions, and there was a statistically significant difference in the scores of Chinese medicine and pharmaceutical majors (P＜ 0.05).Conclusion Teaching experience and learning support need to be optimized for pharmaceutical students.Learning scientificalness and comprehensive abilities in pharmaceutical education need to be improved.Mid and higher level of learning behavior needs to be enhanced for pharmaceutical students.

Objective To develop a liquid bandage for self aid of war wound.Methods Solution A and B were prepared containing functional polymers derived from the biocompatible poly (ethylene glycol) and hyaluronic acid,which had ability of fast crosslinking reaction when they were mixed.Among them,A solution mainly comprised of mercapto group-containing hyaluronic acid derivative (HA-SH),water-solubility starch,propylene glycol,benzoic acid and etc.B solution contained pentaerythritol tetraacrylate derivative (4arm-PEG-Acrylate) and benzoic acid.The structure of mercapto group-containing hyaluronic acid derivative was confirmed by IR,1H-NMR and etc.HA-SH and 4arm-PEG-Acrylate were formulated to different concentrations (W/V) in different buffers.The two kinds of solution were mixed in different ratios,and in situ crosslinking hyaluronic acid hydrogel was obtained;the crosslinking time was recorded.The adhesive force and waterholding capacity of the hydrogel after crosslinking were measured by adding excipients such as water-solubility starch.Results In case the concentration of HA-SH and 4arm-PEG-Acrylate was 2％ and pH value was 11,the hydrogel film on wound skin would be quickly formed in 20 seconds for wound dressing after spraying A and B solution onto the wound skin.Conclusion The liquid bandage has strong adhesive force,high water-holding capacity and controllable crosslinking time,so it could be used as a novel wound dressing for war wound.

Objective To explore the expression and role of LAT 1 in mouse uterus on early placenta formation on day 8 of pregnancy (D8).Methods One hundred and twenty 6-8-week old SPF female Kunming mice were used in this study.Immunohistochemistry was applied to determine the localization of LAT 1 protein in the mouse uterus on D8 of preg-nancy.The ectoplacental cones (EPCs) were dissected out from D8.5 uterus, and then cultured in vitro with different con-centrations of BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, specific antagonist of LAT1) and L-leucine ( substrate of LAT1) to determine the role of LAT1 during the EPC attachment and outgrowth .Results LAT1 protein was highly expressed in secondary decidual zone and also positively expressed in the mouse uterus on D 8.As a specific antago-nist of LAT1, BCH significantly suppressed the ectoplacental cone outgrowth , whereas L-leucine showed no significant effect on it.Conclusions LAT1 is expressed in the mouse uterus during early placenta formation and promotes ectoplacen -tal cone outgrowth , suggesting that LAT1 may promote the trophoblast invasion into maternal decidual tissue , and partici-pates in the early formation of placenta .

Objective To investigate the synergistic effect of 11 β-hydroxysteriod dehydrogenase (11 β-HSD1) and S100A16 on the differentiation of3T3-L1 preadipocytes and its mechanism.Methods Lentiviral vectors PLJM1-11β-HSD1 and PLJM1-S100A16-GFP were respectively constructed and co-transfected into 3T3-L1 preadipocytes.The cell strains expressing 11 β-HSD1/S100A16 were screened with 2.5 μg/ml puromycin for two weeks.Western blot was employed to verify the lentiviral carrier transfection effects.The expressions of marker genes related to the adipocyte differentiation were detected by mean of realtime PCR.Oil red O staining was used to observe the lipid droplet accumulation and the content of triglyceride was measured after differentiation of preadipocytes.The effect of 11β-HSD1 and S100A16 on PPARγ promoter activity was detected by luciferase reporter gene.Results Compared with the empty vector group,the expressions of 11β-HSD1 and S100A16 protein in the lentivirus cotransfected 3T3-L1 cell strain were significantly higher.After 3T3-L1 cell strain co-expressing 1 1β-HSD1 and S100A16 was induced to differentiate for 8 days,the lipid droplets accumulation and triglyceride content were siginificantly increased,along with increased expressions of adipocyte differentiation marker genes such as PPARγ,CCAAT/enhancer binding protein α,lipoprotein lipase,fatty acid synthase,and adipocyte fatty acid-binding protein,in comparison with 11 β-HSD1 or S100A16 overexpression.The result of reporter gene indicated that 11 β-HSD1/ S100A16 enhanced PPARγ promoter activity.Conclusions 11β-HSD1 and S100A16 may jointly promote the differentiation of 3T3-L1 preadipocytes through a synergistic effect on PPARγexpression and play a critical role in the development of obesity.

Objective To evaluate the effect of the serum of rats with hepato-pulmonary syndrome (HPS) on the expression of caveolin-1 and VE-cadherin in pulmonary microvascular endothelial cells (PMVECs).Methods Among the 40 healthy Sprague-Dawley rats,aged 3-4 months,weighing 220-250 g,20 rats were taken randomly for establishment the model of HPS which was produced by chronic ligation of the common bile duct,and the left 20 rats served as sham operation group.Primary PMVECs were harvested from healthy adult Sprague-Dawley rats and inoculated in ECM culture medium or on 96-well culture plate.The PMVECs of 4th-9th generation were randomly divided into 2 groups (n =36 each):control group (group C) and HPS group.In group C,the serum obtained from normal rats in sham operation group was added to PMVECs,while the serum obtained from rats with HPS was added in HPS group.The final concentration of serum was 10％.After being incubated for 12,24 and 36 h (T1-3),the expression of caveolin-1 and VE-cadherin in PMVECs was detected by Western blot,and the PMVEC adhesion rate and proliferation were determined by CKK-8 method.Results Compared with group C,the expression of caveolin-1 and VE-cadherin was significantly down-regulated,the cell adhesion rate was decreased,and the proliferation of PMVECs was enhanced in HPS group.Conclusion The serum of rats with HPS induces weakened PMVEC contact inhibition through down-regulating caveolin-1 and VE-cadherin expression.

Objective To investigate the phenotype and genetic characteristics of a Chinese family with an autosomal-dominant inherited sensorineural hearing loss.Methods A Chinese pedigree associated with an autosomal-dominant inherited low-frequency sensorineural hearing loss (LFSNHL) was investigated.After obtaining informed consent from all study participants medical and audiological examination were used to rule out any syndromic hearing impairment.Five patients were tested with DPOAE and ABR,while two patients were tested with vestibular function and computed tomography scan of the temporal bone to exclude auditory neuropathy and other possible aural disorders.Twenty-one loci and twenty-three genes of DFNA screening had been done by using microsatellite markers and linkage analysis.Results Proband of the family had been diagnosed with low-frequency sensorineural hearing loss.A Chinese family BJ-L046 with non-syndromic autosomal dominant hearing loss was ascertained.Hearing impairment in the affected members in family BJ-L046 occured from 10 to 20 years of age and mainly affected the low frequencies,causing an upsloping audiogram.Higher frequencies were getting involved with increasing age,thus causing a flat-type audiogram.No positive result was found in twenty-one loci and twenty-three genes of DFNA screening.Conclusion A Chinese family with late-onset low-frequency sensorineural hearing loss was clinically studied.No positive result was found by linkage analysis,and twenty-one loci and twenty-three genes of DFNA were preliminary excluded.

Objective To investigate the role of annexin A1 (ANXA1) in human pulmonary arterial smooth muscle cells (PASMCs) proliferation induced by hypoxia.Methods Primary cultured human PASMCs were randomly divided into 5 groups ( n =60 each):normal oxygen group (group N) and hypoxia 2,8,12 and 24 h groups(groups H1-4).Group N was treated with 21％ O2 for 24 h,and groups H1-4 was treated with 3％ O2 for 2,8,12 and 24 h respectively.ANXA1 mRNA and protein expression was determined by RT-PCR and Western blot.Proliferation of PASMCs were determined by MTT and 3 H-TdR methods.Results ANXA1 mRNA and protein expression was down-regulated,proliferation of PASMCs was enhanced in groups H1-4 as compared with group N (P ＜ 0.05).ANXA1 mRNA and protein expression was down-regulated gradually,proliferation of PASMCs was enhanced gradually in groups H1-4 ( P ＜ 0.05 ).Conclusion The expression of ANXA1 is down-regulated in human PASMCs during hypoxia treatment,which might be the mechanism of human PASMCs proliferation induced by hypoxia.