BACKGROUND: Most animal experiments of transgene are derived from mice; therefore, it is necessary to establish a focal cerebral ischemiareperfusion model and significant to prevent and cure ischemic cerebrovascular diseases.OBJECTIVE: To establish a convenient and reliable model with middle cerebral artery occlusion reperfusion (MCAO/R) in mice.DESIGN: Randomized controlled animal study.SETTING: Institute of Cerebrovascular disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: Twenty healthy BALB/c mice, of both genders, weighing 25-30 g, of SPF grade, were divided into sham operation group (n=5), ischemia group (n=10) and 22-hour reperfusion group (n=5) on the basis of digital table. In addition, according to digital table, 130 healthy male Kunming mice were divided into sham operation group (n=10), 24-hour ischemia group (n=30), 2-hour ischemia/22-hour, 46-hour and 70-hour reperfusion groups with 30 in each group; meanwhile, 30 female mice were divided into sham operation group, 24-hour ischemia group and 2-hour ischemia/22-hour reperfusion group with 10 in each group. All Kunming mice were weighing 25-30 g and of SPF grade.METHODS: The experiment was carried out in the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from July 2005 to March 2006. The 6-0 suture with silica gel covered at an end was sent from the left external carotid artery (ECA) into internal carotid artery (ICA) till arriving at the initiation of middle cerebral artery (MCA) to block the blood stream in it, then drawing the suture from ICA 2 hours after occlusion to accomplish reperfusion. Mice were cut off their heads in sham operation group at 24 hours after operation, in ischemia group at 24 hours after blocking blood stream and in reperfusion group at 24, 48 and 72 hours after operation. Reliability of models was evaluated with neurology score and tetrazolium chloride stain. Longa standard scores: neurology score ≥ 1 point was regarded as successful models;coronal sections of brain tissue were stained with tetrazolium chloride, and the white region was regarded as infarcted volume.MAIN OUTCOME MEASURES: Neurology score and infarcted volume after staining of triphenyltetrazolium chloride in brain tissue.RESULTS: All mice were involved in the final analysis. ① Successful rate was 20% of BALB/c mice, 66.7%-73.3% of male Kunming mice and 40%-50% of female Kunming mice. ② Brain sections of BALB/c mice in sham operation group were orange at both sides of cortex and infarction focus was not observed. A big infarcted volume was observed on brain sections of mice in ischemia group, and infarcted volume counted for 50%-70% as homonymy hemisphere on optochiasmatic coronal sections. The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%. In addition, condition in ischemiareperfusion group was similar to that in ischemia group. A big infarcted volume was observed on brain sections, and infarcted volume counted for 50%-75% as homonymy hemisphere on optochiasmatic coronal sections.The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%.CONCLUSION: The model with MCAO/R in mice characterizes by relatively smaller trauma, and the ischemic region is stable; therefore, it can be used to accurate timing control of ischemia/reperfusion. This model is an ideal one for researching pathophysiological changes, prognosis and therapy in cerebrovascular disease.

Objective: To clone the 5＇-flanking region of the human CD34 gene containing transcriptional regulatory sequence (TRS). Methods: According to the registered 5＇-flanking region of CD34 gene, two pairs of primers were designed and net-PCR was used to amplify 661 bp long TRS of CD34 gene. The CD34 TRS fragment was cloned into reported plasmid pEGFP-1. The role of the regulating the specific expression of recombinant plasmid pCD34 EGFP in hematopoietic and non-hematopoietic cells was observed. Results: Restrictive endonuclease identification and DNA sequencing provedthat the CD34 promoter cloned was consistent with the sequence reported to a large extent. It could induce the EGFP gene to express in hematopoietic cell line K562 specifically, while has no effect on hepatocellular carcinoma cell HepG-2. Conclusion: The cloned CD34 gene TRS has the effect of regulating gene expression specifically. The study established the fundament for the construction of specific gene expression vector used in hematopoietic system cells.

Objective To analyze the effect of malaria surveillance and evaluate the surveillance measure in Guangxi Zhuang Autonomous Region from 1995 to 2004 so as to provide scientific basis for formulating control strategy at the late-stage of malaria control. Methods The data on blood smear examination of febrile patients among local residents, focus residents and mobile population in 92 counties from 1995 to 2004 were collected,described and analyzed. Results The totals of 10920395,427600,246159 and 253530 slides in local residents,focus residents, returned population and endemic population were examined. The average positive rates of blood examination were 0.009%,0.079%,1.386% and 0.324%,respectively. The indigenous cases and imported cases accounted for 23.29% (1285/5517) and 76.71% (4232/5517) of the total malaria cases respectively. Conclusions The malaria incidence in Guangxi Zhuang Autonomous Region has been under 1/10000 for 17 years. There is less indigenous malaria in Guangxi. The imported malaria cases are dominated and scattered in whole Region. The secondary cases of imported malaria is very low. The results show that the present surveillance system and control programs are effective for monitoring the epidemic situation of malaria.

Objective To label the amino-glucomannan(AGM) with fluram and to explore the properties of the compound of fluram-AGM in transmembrane transport.Methods After AGM was labeled with fluram,the peripheral blood mononuclear cells(PBMC) and HepG2 were respectively cultured with Fluram-AGM,then observed under fluorescence microscope with violet light as exciting light.Results Both PBMC and HepG2 showed intracellular indigotic fluorescence.Conclusion AGM can be transported into cells across cell membrane.

Objective To study the condensation mechanism of aminoglucomannan(AGM)with plasmid DNA and some influencing factors in this process.Methods The polysaccharide-DNA complex was analyzed with agarose gel electrophoresis by observing the changes of DNA bands under various influencing conditions including different pH,ionic strength(NaCl concentration),temperature and plasmid DNA sorts.Results The new bands appeared in the slow migration positions after reaction between AGM and plasmid DNA,and the changes of DNA bands content were dependent on AGM concentrations.The condensation degree could be effected by pH,temperature,and ionic strength.Conclusion Our study suggests that the AGM can be condensed with plasmid DNA in proper conditions.

Objective:To explore the interaction between gossypol and human serum albumin ( HSA) . Methods:The interaction of gossypol and HSA under physiological conditions was studied by fluorescence spectroscopy, and the molecular docking software was used to simulate the interaction. Results:The binding constant of gossypol and HSA at 293K and 303K was 2. 390 6 × 105 L·mol-1 and 3. 576 8 × 103 L·mol-1 , respectively. There was one binding site on HSA for gossypol. Hydrogen bond and Van Der Waals inter-actions were involved in the binding process. The binding of gossypol and HAS was closer to tyrosine residue in HSA. The molecular simulation analysis verified the above results. Conclusion: The gossypol-induced fluorescence quenching of HSA belongs to a static quenching procedure.

?AIM:To study the clinical value of non-steroidal anti-inflammatory drug in adjuvant treatment of intravitreal triamcinolone acetonide ( IVTA ) for macular edema caused by retinal vein occlusion ( RVO) .?METHODS: Forty - eight eyes in 48 patients were randomly divided into trial and control group ( 24 eyes each ) in this prospective study. In the trial group, additional pranoprofen drops was administered from 1d before IVTA to 30d after injection. Central foveal thickness ( CFT ) was measured with optical coherence tomography ( OCT ) . Available documents of best corrected visual acuity ( BCVA ) , CFT, intraocular pressure and complications pre- and post-injection at 3d, 1,2wk, 1 and 3mo were evaluated.?RESULTS: After IVTA, BCVA was improved in both groups at different levels; but there was no statistically significant between two groups at each time point ( P>0. 05). The CFT values were 629 ± 43μm vs 605 ± 57μm before IVTA in the trail vs control groups (P>0. 05). The values were 432±74μm vs 511±32μm (t=7. 533, P<0. 05), and 275±54μm vs 379±29μm (t=13. 212, P<0. 05) of the trial vs control groups at 1 and 3mo after IVTA, respectively. Ocular hypertension occurred in 5 eyes after injection in trail group, and was controlled with anti-glaucoma medication and one eye with filtration surgery. Progression of cataract was noted in 3 of 35 phakic eyes and cataract surgery was performed in 2 eyes at 4-12mo after injection in trail group. Progression of cataract was noted in 4 eyes and cataract surgery was performed in 2 eyes at 4- 12mo after injection in control group. No retinal detachment and endophthalmitis happened during the whole period of follow-up.?CONCLUSION: Application of non - steroidal anti -inflammatory eye drops in perioperative period can be useful to improve the outcome of IVTA for macular edema, which needs further evaluation.

Objective To evaluate the value of ultrasound in diagnosing ductal carcinoma in situ (DCIS). Methods The sonographic characteristics of 12 DCIS which were confirmed by pathology were analyzed retrospectively. Results The ultrasound image of DCIS could be divided into four types;the solid mass nodule, mammary dysplasia, mix mass nodule, the dilated duct type. Micro calcification had high incidence rate. Ultrasonic diagnosis accordance rate was 50.0 %. On molybdenum target mammograms, the tumor appeared as a cluster of calcified spots in 8 cases, and the accuracy rate of diagnosis of was 66.7 %.Conclusion There are no typical characters of DCIS in ultrasound image. However, some characteristics are suggestive and can help to differentiate them from the benign tumors, such as small nodule, irregular shape,obscure boundary, and microcalcification. When sonography combine with molybdenum target mammography,the accuracy rate of diagnosis will be improved.

Objective:To discuss treatment of complete response cases after neoadjuvant radiotherapy in rectal cancer. Methods:This retrospective study analyzed clinical data of 84 rectal cancer cases with pre-operative neoadjuvant chemoradiotherapy in our hospital from January 2010 to Augnst 2014. Results:After neoadjuvant chemoradiotherapy, 33 patients presented clinically complete response at a rate of 39.3%. After post-operative pathologic examination, among clinically complete response cases, six cases exhibited patho-logically complete responses at a rate of 18.2%. No recurrence or disease progression occurred within 12-36 months of post-operative follow up. Conclusion:Neoadjuvant chemoradiotherapy can significantly lower tumor stage and promote clinically complete remission of some patients. However, for clinically complete remission cases, further radical surgery should be provided.

[Objective] To investigate the effects of the bioactive constituents of green tea polyphenols epigallocatechin gallate (EGCG) on low density lipoprotein receptor (LDLR) function of HepG2 cells.[Methods] The optimal concentration and cell proliferation of HepG2 cells were determined by CCK8 assay,and Western blotting was used to determine LDLR and PCSK9 protein levels,respectively,and LDL uptake in HepG2 cells was detected by fluorescence microscope.[Results] EGCG elevated LDLR protein expression,reduced PCSK9 protein expression and promoted LDL uptake in HepG2 cells.[Conclusion] EGCG may increase LDLR abundance by down-regulating PCSK9 protein and attenuating LDLR protein degradation,which providing a new approach for lipid lowering therapy.