Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

Objective To investigate the application of strain rate imaging in quantitative assessment of left ventricular regional diastolic function after intracoronary stent implantation.Methods Fifty-six healthy person and 60 patients with coronary artery disease were performed quantitative assessment of left ventricular regional diastolic function by apical four chamber view,apical two chamber view and left ventricular long axis view before and after intracoronary stent implantation using strain rate imaging.Results Compared with those of normal myocardium,the peaks of strain rate curve at diastole of ischemic myocardium were reduced,and were increased after operation (P ＜ 0.05).Conclusions Intracoronary stent implantation can significantly improve the blood supply to the ischemia myocardium.Strain rate imaging can quantitatively analyze the changes of left ventricular regional diastolic function.

The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.

The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.
Animals ; Capsules ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Immunologic Factors ; pharmacology ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; drug effects ; immunology

Hepatitis B ; transmission ; virology ; Hepatitis B virus ; Humans ; Infectious Disease Transmission, Vertical

Objective To develop a multiplex PCR-based reverse line blot(mPCR/RLB) hybridization assay to detect,simultaneously,seven genes encoding AR-erm(A/TR),erm(B),mef(A/ E),tet(M),tet(O),aphA-3,aad-6 and two AR-related genes,int-Tn and mreA in group B streptococcus.Methods Nine pairs of specific primers and Oligonucleotide probes targeting erm(A/TR), erm(B),mef(A/E),tet(M),tet(O),aphA-3,aad-6 int-Tn and mreA respectively were modified according to former studies or designed in this study.The primers and probes were labeled with biotin and amino,respectively.The nine genes were amplified simultaneously in the same tube.PCR product hybridized with the probes labeled in the BiodyneC nylon membrane to detect the nine genes.To detect the sensitivity and specificity of the method developed,PCR with single pair of primer targeting each gene were tested in 318 isolates tested and the results were compared with the one abtained by RLB.Results The nine resistance-related genes could be successfully detected by mPCR/RLB assay developed in this study.Based on sequencing,21 of 22 isolates with mef had mef(E)and eight of 353 with int-Tn had an atypical sequence.Except for the above 29 genes,all the others corresponded well with the results obtained by single pair primer PCR.Conclusion The mPCR/RLB assay developed in this study is simple,rapid and suitable for surveillance of antibiotic resistance in GBS.

The efficacy of ~(131)I treatment of hyperthyroidism in children and adolescents was evaluated. Being unsuitable for medical therapy,31 patients (aged 11-18 years) with hyperthyroidism received ~(131)I treatment with a dose of 0.925-3.33 MBq/g of thyroid and were followed-up for 20 to 76 months.Fifteen patients were euthyroid,5 suffered from late-onset hypothyroidism,and 11 were still hyperthyroid,but their symptoms and signs of hyperthyroidism were markedly improved.Of the 18 patients with thyroid-associated ophthalmopathy (TAO),8 patients recovered,4 were improved,TAO in 1 patients deteriorated and in S patients remained unchanged.~(131)I is a relative safe and effective treatment for children and adolescents above 10 years old with hyperthyroidism,being unsuitable for medical therapy.

Heart failure has become a global public health problem that seriously threatens human health. Due to "multi-target" effect, traditional Chinese medicine has unique advantages in the treat-ment of heart failure. Astragaloside Ⅳ is one of the main active ingredients of astragalus membrana-ceus.It has the functions of protecting the heart and neovascularization,anti-inflammation,anti-oxida-tion, regulating energy metabolism, neuroprotection and anti-cancer effects. This article reviews the recent progresses of astragaloside Ⅳ in the treatment of heart failure.Data show that astragaloside Ⅳcan inhibit heart fibrosis,attenuate excessively activation of renin-angiotensin system,increase energy metabolism, promote positive inotropic action, inhibit myocardial cell apoptosis, etc, which are all involved in the protective role of astragaloside Ⅳ in rodents or cellular models of heart failure.Astragalo-side Ⅳ may be a potential active ingredient in traditional Chinese medicine for the treatment of heart failure.

Objective To explore the feasibility of imaging and treatment of cervical cancer xenograft model using 131I mediated by hNIS gene transfection. Methods The cervical cancer xenograft models were established with Hela-NIS( +) cells and Hela cells, respectively. Five Hela-NIS( +) xenograft models and five Hela xenograft models were dynamically imaged at 0.5, 1, 2, 4, 8, 16 and 20 h postinjection of 131I(7.4 MBq). Five Hela-NIS( +) xenograft models were imaged at 0. 5,1,2,4,8,16, 20 and 25 h postinjection of 99TcmO4-(11.1 MBq). Twenty Hela-NIS( +) cervical cancer xenograft models were randomly divided into four groups: Three 131I treating groups and one control group. The therapeutic effects of 131I at threelevels (74,111,148 MBq) were investigated following intraperitoneal injection. Results Hela-NIS( +)human cervical cancer xenografts were established successfully in nude mice. The Hela-NIS( +) xenografts significantly accumulated radioactivity after intraperitoneal injection of 131I, and the radioactivity was persistently present until 20 h postinjection, but Hela xenografts had no radioactive accumulation. The T/B value of the Hela-NIS( +) xenografts reached 17.34 at 8 h postinjection. The imaging with 99TcmO4- showed that the radioactivity was persistently present in Hela-NIS( +) xenografts for almost 25 h. The Hela-NIS( +)xenografts shrinked after 131I treatment. The inhibition ratios of tumor growth in 111 MBq and 148 MBq groups were both significantly higher than that of 74 MBq group (t: 2.74-5.75, P ＜0.05). Conclusions Hela-NIS( +) cervical cancer xenografts in nude mice could persistently accumulate 131I and 99TcmO4- and could be treated successfully with 131 I. 131 I treatment mediated by hNIS gene transfection could be a promising cancer treatment method.

Objective To explore the therapeutic effect and application value of retroperitoneal laparoscopic combined with resectoscopic radical nephroureterectomy for upper tract urothelial carcinoma Methods From Jan.2006 to Jul.2009,fifteen upper tract urothelial carcinoma patients underwent excision of bladder cuff with resectoscope at first,and then retroperitoneal laparoscopic radical nephroureterectomy.All tumors were confirmed to be localized,stage T1-T3.Clinical outcomes of the patients were retrospectively analyzed.Results Mean operative time was 150 (range:120-180) minutes and blood loss volume was 200 (range:100-400)ml.The function of intestinal canal recovered after 24-48 hours,the drainage tube could be removed after 3-4 days.Catheter was kept for 7-10 days.During the follow up for 1-40 months,all the 15 patients survived with one retroperitoneal lymphatic metastasis.There were no severe complications in perioperative and postoperative period.Conclusions Retroperitoneal laparoscopic combined with resectoscopic radical nephroureterectomy may be a practical surgical procedure for upper tract urothelial carcinoma patients with less intraoperative blood loss and early recovery.