Objective:To study the factors affecting amplified fragment length polymorphism(AFLP) of Carthamus tinctorins L.genome,and to develop and optimize the AFLP reaction system,laying a foundation for Marker-assisted breeding of Carthamus tinctorins L..Methods: Cetyl-trimethyl-ammonium-bromide(CTAB) method was used to extract the genomic DNA of Carthamus tinctorins L.,whose concentration and purity were determined by UV spectrophotometry.The genomic DNA was digested by restricted enzyme and ligated with one-step method or two-step method so as to find out which one is more suitable.Then the products were prepared with different dilution multiples(5,10,15,20,25 and 30) for pre-amplification and with different concentrations of gradients(10,25,50,75,100,150 and 200)for selected amplification.Then they were subjected to PAGE electrophoresis and silver-staining after denaturing at 95℃ for 8 min.Results: The AFLP reaction system for Carthamus tinctorins L.genome included the following steps: the genomic DNA was precipitated with alcohol by CTAB method to avoid contamination by any inhibitors;the DNA was completely digested by MseⅠ and EcoRⅠ at 37℃ for 3 hours.Two-step method was superior to one-step method in this AFLP system.The digestion time was 3 hours at 37℃ and ligated at 16℃ over night,with Buffer 2(NEB Company) as the specific buffer.Furthermore,the optimal dilution was 25 folds for pre-amplification and 75 folds for selected amplification. In this reaction system,the strips in PAGE electrophoresis were clear and less decomposed.Conclusion: The AFLP reaction system in this experiment is suitable for silver-stained AFLP of Carthamus tinctorins L.analysis.

Objective:To investigate the effects of Penehyclidinehydrochloride(PHCD) on endotoxin-induced cerebral edema.Methods:one handred and five S-D male rats(200～220g)were randomly divided into 5 groups(n=21):groupC control;groupL LPS;group DL,DM and DH received intraperitoneal PHCD 0.05,0.15 or 0.45mg/kg 10min before lipopolysaccharide(LPS) administration.Cerebral edema was induced by internal carotid arterial LPS 150?g.Seven animals each group were decapitated at 4、6 and 12h after operation and their brains were immediately removed for determination of water content of brain and microscopic examination.Results:Administration of LPS caused severe brain damage and significantly increased water content of the brain.The LPS-induced changes were mitigated by pretreatment with different doses of PHCD in groupDM and DH,but there was no significant difference between the two groups.Conclusion:Pretreatment with PHCD can attenuate LPS-induced cerebral edema.

Sulfur dioxide has been considered as a toxic gas.However,recently more and more studies have shown that sulfur dioxide can be generated endogenously in cardiovascular system and has broad physiologic effects.Studies demonstrate that sulfur dioxide plays an important role in the regulation in the pathophysiologic process of many cardiovascular diseases,such as hypertension,atherosclerosis.These findings suggest that sulfur dioxide might be a novel endogenous gaseous signaling molecule involved in the regulation of cardiovascular functions.

The incidence of bronchial asthma in children is increasing nowadays.More and more studies have showed that epigenetics plays an important role in the pathogenesis of asthma.One of them are histone modification(acetylation and deacetylation),which is catalyzed by histone acetyltransferase (HAT) and histone deacetylase (HDAC).HAT and HDAC can mediate asthma via causing Th1/Th2 imbalance,increased airway hyperresponsiveness,regulation of various inflammatory medium and so on.This paper reviews the HAT and HDAC mediated incidence of asthma and other possible mechanisms,the latest of glucocorticoid in the treatment of children with asthma epigenetic mechanisms.

Objective To investigate the hemodynamic characteristics of superior thyroid artery and its corresponding clinical significance with patients of Graves'hyperthyroidism.Methods Thirty patients with Graves'hyperthyroidism received Doppler ultrasonography.Patients were grouped with different serologic indices at 4 month after radioiodine therapy as relief(R),uncured(U)and hypothyroidism(H).The relationships between thyroid volume(V),superior thyroid artery peak systolic velocity(PSV),end diastolic velocity(EDV),RI,PI and systolic accelerating time(SAT)and the 4-month outcome were analyzed respectively.Results Besides the effect of 24 h-131I uptake rate on patients'short-term outcome,V,PSV and RI were of predominate prognosis value(P＜0.01).PSV and V of group R were positively correlated with 131I dose respectively(r1=0.825,r2=0.697;P＜0.001),while SAT of group R was negatively correlated with 131I dose(r=-0.489,P＜0.05).Conclusions Patients with high V,PSV and RI maybe still suffering hyperthyroidism after regular dosage of 131I therapy.Doses of 131I should be added appropriately for patients with high PSV and V,but short SAT to achieve complete remission,while it should be reduced for patients in opposite condition to avoid occurrence of hypothyroidism.

The primary focal area of carcinoma of unknown primary (CUP) can not be determined by clinical examination.It is difficult to find the primary site and its prognosis is poor.The traditional diagnostic approaches mainly include pathological examination,endoscopy,imaging technology and so on.However,positron emission tomography is the preferred approach.In recent years,gene expression profiling has become a new diagnostic approach to find the source of CUP.The detection rate by gene expression profiling is high,and the approach can accurately identify the primary site.The recommended therapies include classic platinum-or paclitaxel-based chemotherapy,radiotherapy or surgery,and site-directed therapy by gene expression profiling for patients with CUP improves their survival rate.

Department ofPediatrics,the Maternal and Child Health Hospital ofHuzhou,Huzhou,Zhejiang 313000,China(Shen ZZ)Objective To observe the effect of phentolamine in the treatment of severe hand foot and mouth disease with acute pulmonary edema.Methods 90 children with severe hand foot and mouth disease and acute pul-monary edema from January 2014 to December 2015 in our hospital were selected as the research subjects.All the children were randomly divided into 45 case of the observation group and 45 case of the control group.They were given conventional drug therapy and positive pressure ventilation,the control group was given milrinone,5μg/kg,the obser-vation group was given phentolamine,5μg/kg.The myocardial enzymes and cardiac troponin were detected before intervention and after 72h intervention,the blood gases index and ventilator parameter were analyzed,and the time of ventilator support and hospital stay were compared.Results (1)The CK -MB,CK,troponin of the observation group and the control group after treatment were (17.52 ±1.33)U /L and (28.23 ±3.89)U /L,(120.42 ±6.21)U /L and (150.86 ±8.67)U /L,(0.46 ±0.07)μg/L and (0.96 ±0.06)μg/L,which were significantly higher than before treatment (t =5.024 and 8.455,9.353 and 14.757,6.356 and 9.505,all P <0.01),but the CK -MB,CK,troponin of the observation group after treatment were significantly lower than the control group (t =5.601,7.426,5.829,all P <0.01 ).(2)The PaO2 and PaCO2 of the intervention group and the control group after 72h intervention were (80.24 ±14.35)mmHg and (62.30 ±5.68)mmHg,(39.38 ±6.08)mmHg and (46.10 ±3.42)mmHg respectively, which were significantly improved than before treatment,the differences were statistically significant (t =11.352 and 7.204,7.827 and 5.287,all P <0.01),there were statistically significant differences between the two groups after the intervention (t =7.426,5.829,all P <0.01).The FiO2 ,PIP,PEEP of the two groups after treatment were significantly lower than before treatment,the differences were statistically significant (t =8.446 and 6.905,9.705 and 7.832, 8.605 and 6.889,all P <0.01),there were statistically significant differences between the two groups after the inter-vention (t =3.461,5.165,4.532,P <0.05 or P <0.01).(3)The time of mechanical ventilation and hospitalization in the observation group and the control group were (3.42 ±1.61)d and (5.06 ±2.15)d,(15.40 ±3.62)d and (18.62 ±3.81)d,the differences were statistically significant(t =3.513,3.734,all P <0.05).Conclusion Phent-olamine has better effect than milrinone in the treatment of severe hand foot and mouth disease with acute pulmonary edema,it can control the disease progression and improve clinical outcomes and has better clinical value.

Entrepreneurship of the students in TCM colleges can help alleviate the pressure of employment and take advantages of TCM to benefit the public, deepen medical reform, promote the heritage and develop the TCM culture. The influence of the environment could be throughout the whole process of entrepreneurship. The entrepreneurial environment of the students from TCM colleges were analyzed in the following three factors, such as family environment, school environment, and social environment. Only these three environment work together could help entrepreneurship go well. Thus, it is important to change the traditional concept, develop education system of entrepreneurship, and improve the entrepreneurial support system.

OBJECTIVE:To study the effects of different extraction methods on the content of cholalic acid from Artificial Bezoar. METHODS: Artificial Bezoar from 3 different habitats was used to extract cholalic acid by hydrolization vs. hydrolization-free method, and the content of cholalic acid was determined by HPLC-ELSD(high performance liquid chromatography with evaporative light scattering detection). RESULTS: The content of cholalic acid in the hydrolyzed sample was higher than that in the non-hydrolyzed sample of the same batch. CONCLUSION: Through hydrolization, the binding cholalic acid in the artificial bezoar was transformed into free cholalic acid so that the content of cholalic acid was increased. Therefore, it is more reasonable to control the quality of Artificial Bezoar by taking the contents of total cholalic acid and free cholalic acid as indexes.

OBJECTIVE: To analyze the characteristics of adverse drug reaction (ADR) induced by Levofloxacin (LVFX) injection and to promote rational use of drugs in the clinic. METHODS: 137 LVFX injection-induced ADR cases reported in our hospital during 2006～2009 were collected and analyzed statistically with Excel in respect of age, gender, medication and clinical manifestation, etc. RESULTS: ADR cases occurred nearly in all ages of people, and 32.85% were appeared in people over 60 years. Main clinical manifestations were lesion of skin and its appedants (62.04%) and gastrointestinal injury (16.06%). Most of them were mild and medium ADR. CONCLUSION: Great importance should be attached to the rational use of LVFX injection and ADR monitoring to guarantee the safety of patients.