Objective To determine the transfection of PTEN gene in human ovarian carcinoma SKOV3 cells by ultrasound mediating and contrast agent′s enhancing, and to explore the mechanism of PTEN gene inhibiting the tumor cells′ invasion. Methods After addition of PTEN gene and 5 ?l lipofectamine,human ovarian carcinoma SKOV3 cells on 6-well plates were explored to ultrasound for 60 s in the presence(200 ?l) of ultrasound agent. The invasiveness of transfected cells was measured quantitatively by Matrigel invasion assays(Transwell chamber) after 48 h. Results The invasiveness of SKOV3 cells transfected by ultrasound-mediated PTEN gene was significantly declined compared with that of the controls. Conclusions The transfection of PTEN gene into SKOV3 cell can inhibit their invasion.The gene transfection can be enhanced by ultrasound and contrast agent,which may be expected to be a new means of clinical gene therapy.

Objective To research whether double antiplatelet drug combined with pantoprazole can reduce digestive tract damage,or aggravate cardiovascular system side effect.Methods 270 patients diagnosed as ischemic stroke received double antiplatelet drug were randomly divided into two groups.The treatment group received panto-prazole,the control group did not took any gastric drug.The side effect of the digestive and cardiovascular system was observed.Results 268 patients were followed up for 90 days,and digestive system side effect of the treatment group (136 cases)included indigestion in 26 cases(19.11%),stool occult blood(+)in 11 cases(8.08%),melena in 8 cases(5.89%),hematemesis in 0 case,and the control group (132 cases)included indigestion in 42 cases (31.82%),stool occult blood(+)in 23 cases(17.42%),melena in 19 cases(14.39%),hematemesis in 4 cases. There were significant differences between the two groups(χ2 =6.66,4.56,11.2 and 4.18,all P <0.05).Cardiovas-cular system side effect of the treatment group included palpitation or chest distress or chest pain in 4 cases (3.68%),myocardial ischemia diagnosed by electrocardiogram in 4 cases(2.94%),myocardial infarction and died of cardiovascular disease in 0 case,and the control group included palpitation or chest distress or chest pain in 4 cases (3.00%),myocardial ischemia diagnosed by electrocardiogram in 3 cases(2.27%),myocardial infarction and died of cardiovascular disease in 0 case.There were no statistical differences between the two groups(χ2 =6.66 and 4.56, both P >0.05 ).Conclusion Double antiplatelet drug combined with pantoprazole had a positive significance in reducing the digestive tract damage of ischemic stroke patients,and didn't increase the incidence of cardiovascular sys-tem side effect.

Objective To study the molecular mechanism of glutamate receptor-6 (GluR6) in the pathogenesis of epilepsy. Methods Seizure model of SD rats was induced by intraperitoneal injection of kainate (KA). Immunoprecipitation and immunoblotting were performed to examine the interactions of GluR6 and MLK3 with PSD95 at various time points after KA injection. The effect of Tat-GluR6-9c on the MLK3 phospharylation induced by kainate was observed with immunoblotting and immunohistochemistry. Results The assembly of GluR6 and MLK3 with PSD95 was induced after KA hippocampal CA3 region, and bagan to decrease one day later. Pretreatment after KA injection in CA3 region (P<0.05). Conclusion KA induces the assembly of the GluR6-PSD95-MLK3 signaling module and subsequently activates MLK3, which ultimately results in brain injury.

The purpose of this study is to develop glipizide push-pull osmotic pump (PPOP) tablets by using a formulation design expert system and an artificial neural network (ANN). Firstly, the expert system for the formulation design of osmotic pump of poor water-soluble drug was employed to design the formulation of glipizide PPOP, taking the dissolution test results of Glucotrol XL as the goal. Then glipizide PPOP was prepared according to the designed formulations and the in vitro dissolution was carried out. And in vivo evaluation was carried out between the samples which were similar to Glucotrol XL and the Glucotrol XL in Beagle dogs. The range of the factors of formulation and procedure, which could influence the drug release, was optimized using artificial neural network. Finally, the design space was found. It was found that the target formulation which was similar to Glucotrol XL in dissolution test could be obtained in a short period by using the expert system. The samples which were similar to Glucotrol XL were bio-equivalent to the Glucotrol XL in Beagle dogs. The design space of the key parameter coating weight gain was 9.5%-12.0%. It could be concluded that a well controlled product of glipizide PPOP was developed since the dissolution test standard of our product was more strict than that of Glucotrol XL.

Objective:To artificially design and express a recombinant protein named as ScFv-pLLO by fusing ScFv gene of Rituximab(C2B8)and dominant antigen epitopes from listeriolysin O(LLO),and studying its anti-tumor activity.Methods:VH and VL gene sequences of C2B8 against CD20 were acquired by searching United States Patent database,and ScFv sequence was constructed by linking VL and VH with a short peptide linker.Two CD4+T cell epitopes from LLO were selected and designed to splice ScFv sequence.The recombinant gene of ScFv-pLLO was cloned into prokaryotic expression vector and purified after induction.The capacity of ScFv-pLLO target-binding to B-cell lymphomas was evaluated by flow cytometry ( FCM ) and co-immunoprecipitation ( Co-IP ) .The effects of ScFv-pLLO on B-cell lymphomas proliferation and apoptosis were detected respectively.The immunogenicity of ScFv-pLLO was assessed by lymphocyte proliferation assay.Results: ScFv-pLLO was successfully expressed.It could bind to different B-cell lymphomas cell lines and obviously inhibit the growth of Raji cells as well as inducing apoptosis.Moreover,ScFv-pLLO was able to stimulate proliferation of spleen lymphocytes of immunized mice.Conclusion: The recombinant protein ScFv-pLLO can target-bind to B-cell lymphomas,and perform inhibitory effect and induce apoptosis on Raji cells that indicate ScFv-pLLO retain the capacity of ScFv derived from monoclonal antibody against CD20.Besides, ScFv-pLLO can induce immune response.This study provides a basis for further research about the role of ScFv-pLLO on simulating tumor cell antigens as well as being tumor vaccine adjuvant.

OBJECTIVE:To conduct component analysis for the 40％ ethanol eluate of Hyssopus officinalis,and investigate its improvement effect on inflammation in asthmatic mice. METHODS:The 40％ ethanol eluate of polyamide resin column was col-lected,and HPLC-high resolution mass spectrometry was used for the component analysis of 40％ ethanol eluate of H. officinalis. Totally 72 mice were randomly divided into blank group (normal saline),model group (normal saline),dexamethasone group (positive control,1.6 mg/kg) and 40％ ethanol eluate of H. officinalis high-dose,medium-dose,low-dose groups (200,100,50 mg/kg),12 in each group. Except for normal group,mice in other groups were intraperitoneally injected 0.2 mL ovalbumin (OVA)for sensitization in 0,14 d and intragastrically administrated in 25-31 d,once a day. After administration,2 mg/mL OVA was dropped in nose for 7 d. After 24 h of last dropping in nose,tumor necrosis factor α(TNF-α),interleukin-4(IL-4),interfer-on-γ(IFN-γ)levels in bronchoalveolar lavage fluid(BALF)were detected;pathological changes in lung tissue were observed. RE-SULTS:Totally 11 compounds were identified,the relative percentage content of 40.89％. The main components were rosmarinic acid,luteolin 7-O-β-D-rhamnoserhamnose (1→6)-α-D-pyran glucoside,hyperoside,etc. Compared with blank group,TNF-α, IL-4 levels in BALF in model group were increased,IFN-γ level was declined,and IL-4/IFN-γ ratio was enlarged(P<0.01);lung tissue was seriously damaged,there was infiltration of inflammatory cells around the blood vessels. Compared with model group, TNF-α,IL-4 levels in BALF in dexamethasone group,40％ ethanol eluate of H. officinalis high-dose,medium-dose groups were declined,IFN-γ level was increased,and IL-4/IFN-γ ratio was reduced (P<0.01);pathological changes in lung tissue were improved. CONCLUSIONS:The established analysis method can effectively analysis the chemical components of 40％ etha-nol eluate of H. officinalis,which has certain regulatory effect on releasing inflammatory factors and reducing inflammatory lesions in lung tissue of mice with bronchial asthma.

Objective To efficiently builds up and expand breast cancer cells from cancer tissue and to identify their biological properties , provide abundant materials for research and personalized medicine .Methods Feeder cell layer and ROCK inhibitor Y-27632 were employed to faciliate the breast cancer cells;CCK-8 was used to determine the proliferation of the breast cancer cells; Cell cycle distribution was analyzed by flow cytometry; Histochemistry ( FH) assay to show the expression level of CK .The mRNA expression of HER-2, ER, PR and the breast cancer stem cell associated molecules (such as CD44, CD24, etc.) were detected by RT-PCR;STR assay was used for identifying verification of the cells .Results The use of feeder cells and Y-27632 facilitates rapid expand of the original breast cancer cells , and the cells have kept the original features of the tumor .Conclusions To use the method could obtain a large number of cells within a short time , which can promptly be used for the research of per-sonalized medicine .

Objective To investigate the features of the auditory mismatch response (MMR) elicited from the preschoolers and adults.Methods 9 preschoolers aged 3-6 and 8 adults were elicited and measured MMR to speech sounds (/bal/, /pal/) using the Oddball paradigm.Results The response was typical mismatch negativity in adults, and was slow positive waves with larger amplitude in the preschoolers.MNOVA results showed that there were significantly differences between the 2 groups, said the latency of MMRs was significantly longer and the amplitude was larger in the preschoolers than in the adults (P<0.05). Conclusion Stable MMRs with distinct characters in preschoolers and adults have been obtained respectively.

Objective To translate Rheumatoid Arthritis Self-efficacy Scale (RASE) into Chinese and to evaluate its reliability and validity. Methods A total of 188 hospitalized patients with rheumatoid arthritis (RA) were selected as research subjects through convenience sampling method. According to the translation mode of the scale, Chinesization and cultural adjustment were conducted to the English version of RASEto test reliability and validity. Results Item analysis showed that the Chinese version of RASE could discriminate the high-score group from the low-score group (P<0.01). Pearson correlation analysis showed that the correlation coefficient between the score of each item and the total score of the Chinese version of RASE was positively correlated (P<0.01).Exploratory factor analysis extracted a total of 8 common factors, which explained 68.55％of the total variance. The Cronbachαof the Chinese version of RASE was 0.901, and Cronbachαof each dimension ranged from 0.660 to 0.867;the retest reliability was 0.955 after 1 week, and ranged from 0.819 to 0.984 for each dimension;the split-half reliability coefficient was 0.848. Conclusion The Chinese version of RASE has good reliability and validity, which can be applied to the research of self-efficacy of patients with RA in China.

Objective: To explore the characteristics of Auditory Brainstem Response (ABR) in children with ASD, and analyze their relation with the core symptoms of ASD. Methods: Ninty children aged 2-6 with ASD were recruited from Harbin in this study. The data of ABR was collected by using BAEP, and the association among children’s absolute latency and interpeak latency of ABR, core symptoms of ASD children’s behavior and clinical manifestation was analyzed. Results: Compared with the normal average value, children with ASD had longer the absolute latency of wave Ⅰ,Ⅲ,Ⅴ in bilateral ears, which were (1.51±0.20)(3.83±0.34)(5.63±0.23)ms, (1.54±0.16) (3.78±0.23) (5.63±0.22)ms, respectively(P<0.05). Some children’s interpeak latency of Ⅰ-Ⅲ, Ⅲ-Ⅴ, Ⅰ-Ⅴ were longer than normal values. Children younger than 3 years old showed prolonged peak intervals of Ⅰ-Ⅲ and Ⅰ-Ⅴ than children in 3-7 years old. The study has also showed that there was positive correlation between the absolute latency of waveⅠin left ear and the social function defect(r=0.45, P<0.05); there was positive correlation between the latency of wave Ⅴin right ear or the latency of waveⅠin left ear or the Ⅰ-Ⅲ peakinterval and nonverbal communication ability dysfunction(r=0.35, 0.39, 0.34, P<0.05); there was positive correlation between the Ⅰ-Ⅲ peak interval and the repeated stereotyped symptoms(r=0.39, 0.35, P<0.05). Conclusion Children with ASD have abnormal auditory behavior. The absolute latency and interpeak latency of ABR is correlated to some parts of core symptoms of ASD.