Objective To study the role of membrane cytoskeleton linker Ezrin in the growth and metastasis of human breast carcinoma cell line MCF-7.Methods Small interfering RNA(siRNA)targeting Ezrin were transfected into MCF-7 cells.RT-PCR and Western blot were used to detect the down-regulation of Ezrin mRNA and protein.MTT method was used to detect the proliferation of the cells.Flow cytometry was used to detect the cell cycle.Transwell assay was used to detect the migration ability of the transfected MCF-7 cells.Results RT-PCR and Western blot results revealed that Ezrin siRNA down-regulated Ezrin expression at both mRNA and protein levels.Down regulation of Ezrin expression decreased the proliferation rate of the cell line(t=4.479,P＜0.01).The cell proportion in G2-M phase (t=-5.997,P＜0.01)and the invasion ability(t=5.268,P＜0.01)decreased significantly.Conclusion Ezrin plays an important role in the process of proliferation,migration,and invasion of MCF-7 cells.

Objective To study the sensitivity and reliability of rat skin anaphylactoid test method, detect and evaluate the anaphylactoid reaction of traditional Chinese medecine injection. Methods The condition of rat skin anaphylactoid test was optimized by studying the influencing factors of sensitivity and reliability of test with C48/80 as a tool drug, Tween80, endotoxin, China cobra toxin, trichosanthin injection, Tanreqing injection and Xuesaitong injection were investigated. Results The best conditions of rat skin anaphylactoid test was as follows:intrademal inject the drug with insulin syringe, 50-100μL per point, immediately inject 0.5%Evans blue dye 1 mL though caudal vein, 15 min later, kill the rat by carotid artery bleeding, clip dorsal skin to do the test. With this method, Tween80, endotoxin, China cobra toxin and trichosanthin injection all can induce blue stain in rat skin. Tanreqing injection showed no blue stain at the clinical dose. Xuesaitong injection although can induce blue stain in rat skin at the clinical dose, but the results cannot exclude the interference of its pharmacological function. Conclusion The method is simple with short test cycle, less dose of test drug, high detection sensitivity and good reproducibility, but some drug can show false positive result due to its own property.

OBJECTIVE:To investigate the occurrence of ADR induced by over-the-counter(OTC)drug in order to guide the rational use of drugs. METHODS:45 cases of ADR induced by OTC drug were analyzed retrospectively in terms of patient's gen-eral information,allergic history,allergenic drugs,reason for drug use,route of administration,incubation period,days of hospi-talization,clinical manifestation,treatment,outcome,etc. RESULTS:Among 45 patients,there were 14 male and 31 female. The patients were mainly 20-30 year-old(31.1%). Three patients had previous history of allergy(6.7%). Most ADR were induced by oral drugs(82.2%). Reasons for drug use were mainly disease of respiratory system(23 cases,51.1%). The results of causality as-sessment were positive in 3 patients(6.7%),probable in 35(n=77.8%),and possible in 7 patients(15.6%). ADR were mainly induced by non-steroidal anti-inflammatory drugs (NSAIDs,57.8%),Chinese patent medicine (20.0%),drugs for external use (17.8%). The clinical manifestations mainly included exanthematous drug eruption (33.3%),erythama multiforme drug eruption (24.4%),urticarial drug eruption(13.3%). CONCLUSIONS:NSAIDs are the main allergenic drugs,and ADR induced by OTC traditional Chinese medicine can not be ignored. To improve medical knowledge and skills of the public and the scientific use of OTC drug are beneficial to improve the clinical treatment level.

Objective To compare the efficacy and complications of the two surgical methods (between type Ⅰ hysterectomy and type Ⅱ hysterectomy),and to explore the feasibility of type Ⅰ hysterectomy in stage ⅠA cervical cancer.Methods The study group,92 cases(48 cases of stage ⅠA1,44 cases of stage ⅠA2) were performed with type Ⅰ hysterectomy plus selective pelvic lymph node dissection;the control group,93 cases (49 cases of stage ⅠA1,44 cases of ⅠA2) were performed with type Ⅱ hysterectomy plus selective pelvic lymph node dissection.Results The survival rate of 5 years and 10 years in study group were 100 ％ (92/92),100 ％ (74/74) and that in control group were 100 ％ (93/93),100 ％(66/66),respectively.There were no signicant difference between the two group (both P ＞ 0.05).When compared with the control group,the urinary tract infection of the study group was significantly reduced (0 versus 13.99 ％,P ＜ 0.05).Moreover,there were a shorter surgical duration [(96.14±17.20) min vs (116.82±16.30) min].The hemorrhage [(117.35±39.61) ml] and blood transfusion (0 ml) in study group was less common than those in control group [(201.74±46.25) ml,(82.07±16.32) ml] (all P ＜ 0.01).Conclusion There are no difference of 5-year and l0-year survival rate in stage ⅠA patients with type Ⅰ or type Ⅱ hysterectomy,however,the rate of the postoperative urinary tract infection in the former is lower than that in the latter,and also there are a shorter surgical duration,less hemorrhage and reduced blood transfusion requirements in study group.Therefore,type Ⅰ hysterectomy can be effective and applicable for the patients of stage ⅠA cervical cancer.

Objective To report the clinical and genetic characteristics of a dentatorubralpallidoluysian atrophy (DRPLA) pedigree with an onset of cognitive impairment.Methods Clinical data of this pedigree was collected.The numbers of CAG repeats in the exon 5 of atrophin-1 (ATN1) gene were analysed in the proband and the other 4 healthy family individuals.The polymerase chain reaction (PCR) products of the proband underwent cloning-sequencing using an original TA cloning kit.Results There were 5 patients in this family,4 with onset in adult and one in childhood.The proband had an onset manifestation of cognitive impairment,while the other 3 adult patients presented with ataxia.The two-year-old child in the pedigree had myoclonic epilepsy.The proband had 61 CAG repeats in the exon 5 of ATN1 gene.After TA cloning-sequencing of the proband ' s PCR products,there were 2 different numbers of CAG repeats,including 61 and 64.Conclusions The clinical manifestations of DRPLA can have obvious heterogeneity in one family.Some patients present with cognitive impairment.It is very important to test the numbers of CAG repeats of ATN1 gen for DRPLA diagnosis.Somatic mosaicism may be also observed in Chinese DRPLA patients.

Objective To explore a medical insurance model for rare diseases and orphan drugs applicable in China.Methods Eleven rare diseases from six disease systems and thirty-five orphan drugs covered were pinpointed for comparative analysis of their present coverage in various medical insurance systems in China.Results The coverage of 35 orphan drugs in the national essential drug list(2009)and the national essential drug list(2012)is 0 and 37.1％ respectively; the coverage in the national essential insurance medicine list is 54.3％ (19 types),and the average coverage in the medicine list for the New Rural Cooperative Medical Insurance Scheme is 33.6％.The study found insufficient rare diseases included in the Basic Medical Insurance Schemes in general.Conclusion The coverage of basic medical insurance for rare disease patients is low in China.In this regard,such measures should be taken to mitigate the economic burden for these patients,namely expanding the coverage of reimbursement list of orphan drugs,building the special reimbursement system of rare diseases and orphan drugs and improving the medical rescue system for rare diseases in China and their medical care level.

Obj cetive A large number of recent studies show that sevoflurane anesthesia may cause learning and memory dysfunction.The aim of this study was to explore changes of learning and memory ability and hippocampal volume in infantile rats after neonatal interrupted and repeated inhalation of 2.6% sevoflurane through detecting the learning and memory ability by Morris water maze and the hippocampus volume by MRI.Method s Thirty two neonatal SD rats were randomly devided into two groups (n=16):experimental group and control group.Rats inhalated 2.6%sevoflurane in the experimental group and 1 L/min O2 +1 L/min Air in the control group at the postnatal days of 7, 14 and 21 (P7, P14, P21). The learning and memory ability was determined by the Morris water maze test from P31 to P37;The brains of rats were scanned by mag-netic resonance imaging ( MRI) machine under anesthesia with 1%sodium pentobarbital at P37, and the brain and bilateral hippocampal volumes were measured. Results ①In the place navigation test, the escape latency had no significant difference between the two groups (P>0.05).In the spatial probe test, the dwelling time, movement distance and number of entering times in platform quadrant decreased slightly in experimental group compared with those in the control group, while there was no significant difference (P>0.05).②The brain volume [(1.53 ±0.18) cm3 vs (1.60 ±0.13) cm3] and right hippocampal volume [(16.15 ±1.76)mm3 vs(16.46 ±1.71)mm3] had no significant difference between the two groups (P>0.05).The left hippocampal volume [(16.46 ±1.71)mm3] was decreased in the experimental group compared with the control group [(18.10 ±2.53)mm3](P<0.05). Conclusion The learning and memory ability has no significant changes in in-fantile rats after neonatal interrupted and repeated sevoflurane inhalation and MRI examination of hippocampal volume is not sufficient for the diagnosis of cognitive dysfunction.

Objective To explore the relationship between health-promoting lifestyle and work engagement in nurses of traditional Chinese medicine hospitals. Methods Totally 527 nurses from 3A level hospitals of the first, second, third affiliated to Zhejiang University of Traditional Chinese Medicine were investigated by the Chinese revision of Health-Promoting Lifestyle Scale, Chinese version of Utrecht Work Engagement Scale. Results M ultiple regression analysis showed that nurse health-promoting lifestyle had a significant correlation with work engagement, F = 8.987, P < 0.01. Among them, the self-realization dimension to the nurse work engagement most affected, t = 3.287, P < 0.01. Conclusions Improving nurses′health-promoting lifestyle can raise the level of work engagement and stabilize nursing staff and improve quality of care in traditional Chinese medicine hospitals.

Objective: To observe the effect on Tangzu Heji on diabetic foot rat with blood stasis syndrome and to investigate the mechanism in it. Methods: Combination of streptozocin injection and reduction of ambient temperature was used to duplicate diabetic foot rat with blood stasis syndrome. 60 Male SD rats were randomly divided into 6 groups as normal control group, the low temperature control group, diabetic foot group (model group), three groups of Tangzu Heji treatment with high, medium and low dose respectively. In the first three groups the rats were given normal saline, and in the latter three treatment groups the model rats were given Tangzu Heji as 60gkg, 30g/kg, 15g/kg weight, respectively. After two weeks of treatment, the changes of the weight, water intake, blood glucose, blood rheology, acral gangrene symptoms, Choroid blood vessels of the sublingual and ear, serum vascular endothelial growth factor changes were observed. Results: After two weeks of treatment with Tangzu Heji, compared with the model group, a marked increase in body weight, decrease of water intake were happened to the rats in treatment groups. Erythrocyte aggregation index in the low-dose treatment group decreased significantly (P

Methylmalonic aciduria (MMA) is a common inherited autosomal recessive disorder resulting from defects in the enzyme methylmalonyl CoA mutase (MCM, mut complementation group) or in the synthesis of the MCM cofactor adenosylcobalamin (cbl complementation groups). The defects in the mut complementation group accounts for the largest number of patients with isolated MMA. At least 200 mutations in the MUT gene on chromosome 6p12 have been identified in MMA patients until now. This study aimed to investigate the clinical characteristics of MMA and genomic variations in the MUT gene of Chinese patients. Genomic DNA was extracted from 18 patients who were diagnosed as having isolated MMA by gas chromatography/mass spectrometry (GC-MS), and from some of their parents as well. Amplification and direct sequencing of the MUT coding regions (exon 2-13) and their adjacent intronic consensus splice sites were performed in order to identify the disease causing mutations. In this group, six novel mutations in the MUT gene, c.424A>G (p.T142A), c.786T>G (p.S262R), c.808G>C (p.G270R), c.1323_1324insA, c.1445-1G>A and c.1676+77A>C were identified. p.T142A and p.G270R were respectively detected at a heterozygous level in one patient. Two previously reported mutations, c.682C>T (p.R228X) and c.323G>A (p.R108H) were also found in this study. In addition, six previously described single nucleotide polymorphism (SNP), c.636A>G (p.K212K), c.1495G>A (p.A499T), c.1595A>G (p.H532R), c.1992G>A (p.A664A), c.2011G>A (p.V671I) and c.1677-53A>G were identified. In this study, we updated the spectrum of MUT mutations and identified the main MMA-causing mutations in Chinese MMA patients.