Objective To construct the eukaryotic expression vector for mouse microRNA miR-21 and identification its expression activ-ity in 293 cells. Methods The genomic sequence containing pre-miR-21 was amplified from mouse genomic DNA by PCR and cloned into the pRC/CMV plasmid. The constructed recombinant plasmid pRC/CMV-mmu-miR-21 was transfected to 293 cells by lipofectamine 2000, and the stably transfected cells were screened with G418,from which total RNA was extracted for detecting the expression of mature miR-21 by northern blot. In the meantime,a luciferase report plasmid examing the activity of miR-21 named pmiR-21-Luc reporter was also construc-ted,and luciferase activity analysis indicated the product of pRC/CMV-mmu-miR-21 indeed had biological activity. Results Both restriction enzyme digestion analysis and sequencing proved the recombinant plasmids were constructed correctly. The miR-21 was highly expressed in the screened clones of 293 cells and it had good biological activity. Conclusion The eukaryotic expression plasmid of mouse miR-21 was successfully constructed,which laid the foundation of further investigation of the role of miR-21 during skin wound healing.

Objective To study the characteristics of immune function,Epstein-Barr virus(EBV) antibodies and EBV-DNA in children with different clinical types of EBV infection,which provide basis for prevention and treatment of EBV infection.Methods Clinical data of 103 patients suffering from EBV infection were retrospectively analyzed in Xijing Hospital of the Fourth Military Medical University.A total of 103 children were divided into infectious mononualeosis(IM) group(n=68),chronic active Epstein-Barr virus infection(CAEBV) group(n=13) and Epstein-Barr virus-related hemophagocytic lymphohistiocytosis(EBV-HLH) group(n=22).The changes of EBV antibodies,EBV-DNA,immunoglobulin levels,lymphocyte subpopulation and complement series were detected and compared among the three groups.A total of 26 healthy children at the same stage were enrolled as a control group,immunoglobulin levels,lymphocyte subpopulation and complement series were detected in control group,then compared with the rest of the three groups.Results The levels of C3 and C4 in CAEBV group and EBV-HLH group were significantly decreased than those in the control group(P<0.05).The levels of IgA,IgG and IgE in EBV-HLH group,CAEBV group,IM group and control group gradually increased(P<0.05,respectively).The levels of IgA,IgG and IgE in EBV-HLH group and CAEBV group significantly decreased than those in control group(P<0.05,respectively).The levels of IgA,IgG and IgE in IM group decreased than those in control group,while there were no statistically significant differences(P>0.05).CD8+T cells in IM group significantly increased than those in the rest of the three groups(P<0.05,respectively).T cells,CD8+T cells,CD4+ T cells,CD4+/CD8+ ratios,NK cells,B cells of EBV-HLH group significantly decreased than those in the rest of the three groups(P<0.05).The positive rates of EBV antibodies in CAEBV group and IM group were significantly higher than those in EBV-HLH group(P<0.05).EBV-DNA in EBV-HLH group were significantly higher than those of CAEBV group and IM group(P<0.05).Conclusion EBV-DNA levels in the serum are positively correlated with disease types and severity,the pathogenesis of IM,CAEBV and EBV-HLH induced by EBV infection are associated with immune dysfunction.Dynamic monitoring of EBV load and cell immune function can reflect disease status and progress risk.

AIM: To study the relationship between respiratory syncytial virus (RSV) infection and apoptosis, between RSV infection and expressions of FasL, Fas, Bcl-2 and Bax. METHODS: Apoptotic cells were examined by flow cytometry and transmission electron microscope. Immunohistochemical analysis was used to detect the expressions of apoptosis-associated gene FasL, Fas, Bcl-2 and Bax in A549 cells during RSV infection. RESULTS: Apoptotic index increased at 72 h and 120 h postinfection. Apoptotic cells were detected by transmission electron microscope. High-expressions of FasL, Fas and Bax genes and low-expression of Bcl-2 gene were detected by immunohistochemical staining. CONCLUSION: Apoptosis in A549 cells was induced by RSV infection. This apoptosis may be induced by up-regulating the expression of FasL, Fas, Bax genes and down-regulating the expression of Bcl-2 gene.

Objective To evaluate the applied values of susceptibility weighted imaging (SWI) in acute ischemic stroke.Methods Totally 40 patients with acute ischemic stroke were included within 72 hours after symptom onset.They adopted multimodal magnetic resonance imaging examination,including regular magnetic resonance imaging sequence (T1WI,T2WI),time of flight MR angiography (TOF-MRA),diffusion weighted imaging (DWI) and susceptibility weighted imaging (SWI).Furthermore,12 of the 40 patients received dynamic susceptibility contrast enhanced perfusion weighted imaging (DSCPWI) examination.Results Of the 40 patients,12 ones had positive results for multiple lesions of cerebral microbleeds (CMBs) by SWI,and the detection rate was 30％.There were 24 patients showed susceptibility vessel sign (SVS) in SWI,and the susceptibility rate was 60％.In the 12 patients received DSC-PWI examination simultaneously,there were 7 cases had the anomaly range by SWI larger than that by DWI while similar to that by DSC-PWI.Conclusion SWI can show SVS and CMBs clearly,estimate ischemic penumbra,know intracranial conditions completely and supply reliable basis for individualized treatment.

Objective To evaluate the clinical efficacy of Qilinxintongshu pill for treatment of critical patients with unstable angina pectoris (UAP) accompanied by upper alimentary tract damage.Methods A prospective randomized controlled trial was conducted. A total of 60 critical outpatients or inpatients with confirmed diagnosis of UAP accompanied by upper alimentary tract damage in Cardiology Department of Hospital of Traditional Chinese Medicine of Qionghai City from November 2011 to January 2015 were enrolled in the study, and they were assigned to a therapy group (30 cases) and a control group (30 cases) by table of random number. The conventional medical basic therapy was given to both groups, in addition, the patients in therapy group took Qilinxintongshu pill (made of notoginseng, dragon's blood, fruit of immature citron, etc.) 5 g once, three times a day, and the patients in control group orally administered clopidogrel 75 mg once a day, the therapeutic course being consecutive 30 days. After treatment, the rates of main cardiovascular events [including death, a newly-happened myocardial infarction (MI), and obstinate ischemia] and events of alimentary damage were compared between the two groups, and adverse effects were observed.Results There was no statistically significant difference in incidence of cardiovascular events between the therapy group and control group [6.7% (2/30) vs. 16.7% (5/30),P > 0.05]. The incidence of massive hemorrhage of gastrointestinal tract was significantly lower in therapy group than that in control group [10.0% (3/30) vs. 36.7% (11/30),P < 0.05]. In the treatment process, there was no significant untoward side effect, and no abnormalities in routine blood and urine tests, liver and renal functions were found.Conclusion Qilinxintongshu pill for treatment of critical patients with UAP accompanied by upper alimentary damage is safe and effective, and does not enhance the incidence of massive hemorrhage of gastrointestinal tract.

The protective effects of Da Chai Hu Granules (DCHKL) on islet cells which were incubated with 4 mmol x L(-1) alloxan (AXN) were studied. The viability of islet cells were measured with MTT. Insulin released into medium and in islets was detected by radioimmunoassay. Cell apoptosis rate was determined by flow cytometry. The expression of anti-apoptotic gene Bcl-2 and pro-apoptotic gene Bax in islet cells were measured with RT-PCR (reverse transcription polymerase chain reaction). Serum containing DCHKL can promote the activity of islet cells significantly (P < 0.01). Basal insulin secretion and high glucose-stimulated insulin secretion increased significantly (P < 0.01). Serum containing DCHKL can inhibit apoptosis of islet cells, the ratio of apoptosis was decreased. Serum containing DCHKL increased expression of Bcl-2 mRNA and decreased expression of Bax mRNA. DCHKL can significantly promote proliferation of islet cells and increase the amount of basal secretion of pancreatic islet cells and high glucose-stimulated insulin secretion. The expression of Bcl-2 increased significantly. The expression of Bax decreased significantly. DCHKL have a protective effect on the islet cells.

Objective To observe the therapeutic effect of Qilin Xintongshu pill for treatment of patients with non-ST segment elevation acute coronary syndromes(NSTE-ACS)accompanied by gastrointestinal bleeding (GIB). Methods A prospective randomized controlled trial was conducted. A total of 67 hospitalized patients in Cardiovascular Department of Qionghai City Hospital of Traditional Chinese Medicine were divided into therapy group(35 cases)and control group(32 cases). Both groups were given conventional medical treatment(except anti-coagulative agent). In the therapy group,the patients received Qilin Xintongshu pill(the ingredients consisting of panax pseudo-ginsen,dragon's blood,immature orange fruit,etc.)oral administration,once 5 mg and 3 times a day,while in the control group,the patients accepted clopidogrel orally,once 75 g and once a day,the therapeutic course in both groups being 30 days. The major cardiovascular events(such as death,a newly-happened myocardial infarction(MI),MI secondary,obstinately ischemia)and incidence of massive hemorrhage of gastrointestinal tract were compared between the two groups,and the adverse events were observed. Results After treatment,the incidence of cardiovascular events and massive hemorrhage of gastrointestinal tract in the therapy group were decreased significantly compared to those in the control group 〔incidence of cardiovascular events:8.57%(3/35)vs. 28.13%(9/32), incidence of massive hemorrhage of gastrointestinal tract:2.86%(1/35)vs. 21.88%(7/32),both P<0.05〕. Adverse events were not observed in both groups,and the examinations of blood,urine,liver and renal functions were of no abnormalities before and after treatment. Conclusion Qilin Xintongshu pill can effectively reduce the incidences of cardiovascular events and massive hemorrhage of gastrointestinal tract in patients with NSTE-ACS accompanied by GIB in the therapeutic course of 30 days,therefore this traditional Chinese herbal medicine is an ideal agent for treatment of such disease.

Objective:To evaluate the effect of presurgical nasal mode(PNM)combined with nasal diorthosis in the treatment of na-sal deformity and incomplete unilateral cleft lip(IUCL)in infants.Methods:35 infants with IUCL were treated by PNM followed by nasal diorthosis and cheiloplasty.The nasal asymmetry was analysed by measurments of nostrils height,nostrils width and nasal colu-mella angle skewness on the photographs at the initial visit(T0),pre-operation(T1),1 week after operation(T2),1 month after opera-tion(T3)and a year after operation(T4).The other 35 infants with IUCL without PNM treatment were served as the controls.Re-sults:Compared with the controls,the symmetry of nostrils height,nostrils width,nasal columella angle skewness in PNM treated children were significantly improved at T0-T1 and T1-T2(P ＜0.05).there was no significant difference at T2-T3 and T3-T4(P >0. 05).Conclusion:Nasal asymmetry can be improved by presurgical nasal mode treatment followed by preliminary nasal deformity di-orthosis and cheiloplasty.

Objective Fructus Amomi(Sharen) is derived from the dry ripe fruit of Amomum villosum Lour., A.villosum Lour. var. xanthioides T.L. Wu et Senjen and A.longiligulate T.L.Wu, which is widely utilized for its clinic effects on digestive system. However, Fructus Amomi from different species and habitats, possessing different quality, is difficult to identify. In this study, we aim to develop a simple, rapid and reliable method for authentication of Fructus Amomi. Methods Twenty-five batches of samples of Fructus Amomi were collected and electronic nose was introduced into analyzing their odor with multiple mathematical statistics methods. Na?ve bayes network (NBN), radical basis function (RBF) and random forest (RF) were applied to establish different classifiers while BestFirst+CfsSubsetEval (BC) was used to screen the attributes for searching sensor array with higher contributions. Results Firstly, after attribute-screening via BC, the established discriminative models via NBN, RBF and RF could successfully identify genuine and non-genuine samples, presenting correct judging ratios of 78% and 84% through ten-fold cross validation and external test set validation, respectively. Besides, quantity predictive models were constructed as well. In case of content of bornyl acetate, one of the effective components in Fructus Amomi, values were higher than 3.5 mg/g and lower than 1.8 mg/g with sensor response of 0.04 and 0.03, respectively. Conclusion In this paper, quality discriminative model and quantity predictive model of Fructus Amomi were established via electronic nose and multiple mathematical statistics methods. It indicates that electronic nose could be a promising method for quality evaluation of Chinese material medica.