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 research the affection and problem of Curriculum integration teaching method in eight-year term Medical Education.Methods Combination teaching and tradition teaching methods were employed in 100 students (Eight-year-term) and 127 students (Seven-year-term) respectively.A questionnaire survey and teaching effect were analyzed at last (employx2 analysis).Results Currculum integration is more acceptable(x2 =3.92,P ＜ 0.05),effective (x2 =11.07,P ＜0.01 ),systematic (x2 =11.82,P ＜ 0.01 ) and improve self-study ability better(x2 =8.51,P ＜ 0.01 ).But it's not helpful for the study enthusiasm(x2 =0.90,P ＞0.05).Conclusion Combination teaching method is more acceptable and effective than tradition method.However,the enthusiasm of study is not improved obviously.Combination Curriculum integration teaching method is a good one and worth popularizing.

Objective To choose proper proton magnetic resonance spectroscopy(~1XH-MRS) parameters to fit for practical femoral marrow cavity and to produce short-timed,well-repeated and excellent ~1H-MRS images.Methods The tentative study of ~1H-MRS on the normal femoral bone marrow in 26 volunteers was performed with a 1.5 T MR after the informed consent.The single-voxel spectroscopy and stimulated echo acquisition mode were used for ~1H-MRS collection.~1H-MRS parameters for 12 volunteers were 128 acquisitions,1 cm?1 cm?1 cm volume of interest(VOI)size and repeatedly 2—3 times within the same location.~1H-MRS parameters for another:14 volunteers were different numbers of acquisition (128 and 256 times,respectively)and different VOI sizes(2 cm?2 cm?2 cm and 1 cm?1 cm?1 cm, respectively).Results For ~1H-MRS with 1 cm?1 cm?1 cm size of VOI and 128 times of acquisition with the full width haft max of water≤8—12 Hz,the base-line was steady and the signal-noise ratio was high up to 11.31.~1H-MRS was different in the different femoral locations showing the maximum peak sites at near 0.90 ppm(?10~(-6))or 1.65 ppm,but~1H-MRS within the same location was always same or similar with different VOI sizes(1 cm?1 cm?1 cm or 2 cm?2 cm?2 cm)or different numbers of acquisition(128 or 256 times).~1H-MRS acquisition time was not related with the size of VOI but with the numbers of acquisition.128 and 256 times of acquisition cost 199 s and 391 s,respectively.Conclusion With the technique of small size of VOI(1 cm?1 cm?1 cm)and decreased numbers of acquisition(128 times),it is propable to get well-repeated and excellent ~1H-MRS within less time.It is also more practical for clinics to achieve ~1H-MRS of the femoral marrow with the proper technique.

Leaves of Chrysanthemum morifolium were potential medicinal resource. The present study aims to estimate the main bioactive components: total flavonoids (TF), galuteolin (GA), quercitrin (QU), chlorogenic acid (CA) and 3 ,5-O-caffeoylquinic acid ( CQ), which were considered to be the main effective components, in leaves of C. morfolium cultivars in China. The TF content was estimated hy UV-VIS spectrophotometry, while GA, QU, CA, and CQ were quantitatively determined by HPLC. The highest TF content (7. 13% w/w) was found in cultivar Wan Cong (Shexian county). Cultivar Da Bo ( Bozhou county) had the highest GA content (33. 45 mg - g-1); Cultivar Hong Xin (Sheyang county) contained the highest QU content (29.25 mg · g(-1)); Cultivar Chang Ban (Sheyang county) had the highest CA content (13.14 mg ·(-1)). The maximum CQ content (7.35 mg · g(-1)) was observed in culti- r Da Yang ( Tongxiang county). Different cultivars of C. morfolium had significant difference in components, but the leaf and capitulum of C. morifolium. were found to possess similar chemical compositions. The high content of bioactive components in several cultivars suggested the potential utilization of C. morifolium leaves.
China ; Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; growth & development ; Drugs, Chinese Herbal ; analysis ; Plant Leaves ; chemistry

The study is aimed to ensure the quality and safety of medicinal plants by using ITS2 DNA barcode technology to identify Corydalis boweri, Meconopsis horridula and their close related species. The DNA of 13 herb samples including C. boweri and M. horridula from Lhasa of Tibet was extracted, ITS PCR were amplified and sequenced. Both assembled and web downloaded 71 ITS2 sequences were removed of 5. 8S and 28S. Multiple sequence alignment was completed and the intraspecific and interspecific genetic distances were calculated by MEGA 5.0, while the neighbor-joining phylogenetic trees were constructed. We also predicted the ITS2 secondary structure of C. boweri, M. horridula and their close related species. The results showed that ITS2 as DNA barcode was able to identify C. boweri, M. horridula as well as well as their close related species effectively. The established based on ITS2 barcode method provides the regular and safe detection technology for identification of C. boweri, M. horridula and their close related species, adulterants and counterfeits, in order to ensure their quality control, safe medication, reasonable development and utilization.
Base Sequence ; China ; Corydalis ; chemistry ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Molecular Sequence Data ; Nucleic Acid Conformation ; Papaveraceae ; chemistry ; classification ; genetics ; Phylogeny ; Plants, Medicinal ; chemistry ; classification ; genetics

Clinical reports on cardiac syndrome X (CSX) have been increasing in recent years. In general, CSX does not increase the cardiovascular mortality, but it can affect the patient's quality of life (QOL) and increase the incidence rates of cardiovascular and cerebrovascular events. Although a variety of drugs and therapies have been utilized in the clinical treatment, the management of CSX still represents a major challenge due to its unclear pathogenesis. It is necessary to explore more effective treatment programs. Many attempts have been made on trials of the Chinese medicine (CM) treatment for CSX and proved that CM has a certain advantage in efficacy to improve clinical symptoms and QOL. CM may provide a new approach for the effective treatment of CSX.
Humans ; Integrative Medicine ; Medicine, Chinese Traditional ; Metabolic Syndrome ; physiopathology ; therapy ; Quality of Life

Objective To evaluate the situation of musculoskeletal imaging research at home and abroad and the disparity and consistency between them.Methods Statistic analysis was done with the musculoskeletal articles published in those journals during the last decade,including Chinese Journal of Radiology,Journal of Clinical Radiology,Radiology,American Journal of Roentgenology(AJR)and the British Journal of Radiology(BJR).Results The development of museuloskeletal imaging research was very imbalaneed among different areas and hospitals in China.The rate of clinical research treatise and imaging technique articles at home was 76.3%(357/468)and 10.0%(47/468),respectively,while that was 75.7%(731/966)and 10.8%(105/966),respectively abroad.The rate of MR image technique articles in China was lower than that abroad.The rate of articles on basic research at home and abroad was 5.3%(25/468)and 7.9%(76/966),respectively.The rate of articles about museuloskeletal imaging simply using CT or MR at home was 20.1%(94/468)and 29.7%(139/468),respectively,while that was 5.1%(49/966)and 60.4%(548/966),respectively abroad.The rate of articles about interventional therapy and synthetic imaging at home was 9.2%(43/468)and 29.3%(137/468),respectively,while that was 4.7%(45/966)and 22.5%(217/966),respectively abroad.With regard to the diseases category,the research focused on the bone tumors and the tumor-like disease in China,while in other countries,the research focus had been diverted to the trauma of bone and joint,and the anatomy and disease of muscle and ligament,which was unsatisfactory in China.More focus had been put on imaging anatomy, cartilage disease and osteoporosis in other countries than in China.The research on the radiation dosage the subject received in the examination,the apphcation of Micro-CT and Micro-MR to musculoskeletal imaging was developed in other countries,but it had not been reported in China.Conclusion Although great success has been achieved in the research of musculoskeletal imaging in the last decade,there is still much disparity comparing with other countries,and the research is very imbalanced among different areas and hospitals in China.We should emphasize the application of MRI to musculoskeletal system and pay more attention to the basic research and imaging anatomy research,especially the research on ligament,muscle and cartilage.

OBJECTIVETo investigate the possibility of Hantavirus (HV) and Orientia tsutsugamushi (Ot) coinfection in their hosts.

METHODSHV and Ot were used to infect Vero E6 cells cultured in vitro singly, simultaneously or successively. Genes of HV and Ot were identified in different generation cells with RT-PCR.

RESULTSFive experiment groups of infected Vero E6 cells were tested, the results were as follows: HV and Ot were both positive in infected Vero E6 cells passaged 2 times and the positive rate increased following the passaged times in HV and Ot infection groups, simultaneously or successively. However, in the groups which were infected with HV and Ot separately, the gene of HV or Ot could be detected in infected Vero E6 cells passaged only once and the positive rate increased following the times of the passaged. The positive rate was higher in the singly infected groups than in those infected simultaneously or successively.

CONCLUSIONCoinfection of HV and Ot did exist in the hosts while HV and Ot could inhibit each other in the initial infection stage.

Animals ; Cell Division ; Cercopithecus aethiops ; Hantavirus ; pathogenicity ; Hantavirus Infections ; Orientia tsutsugamushi ; pathogenicity ; Reverse Transcriptase Polymerase Chain Reaction ; Scrub Typhus ; Vero Cells

OBJECTIVEto investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.

METHODSfifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.

RESULTScompared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.

CONCLUSIONSin patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.

APACHE ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Prognosis ; Prospective Studies ; Sepsis ; drug therapy ; etiology ; immunology ; Somatostatin ; therapeutic use ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate the anti-angiogenic effects of Pien Tze Huang in vivo and in vitro.

METHODSHuman umbilical vein endothelial cells (HUVECs) were treated with 0 mg/mL, 0.25 mg/mL, 0.5 mg/mL, and 1 mg/mL of PZH for 24 h, 48 h and 72 h, respectively. Chicken embryo chorioallantoic membrane (CAM) model was used to evaluate in vivo angiogenesis. An ECMatrix gel system was used to evaluate in vitro angiogenesis by examining the tube formation of HUVECs. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine HUVEC viability. Cell density of HUVECs was observed by phase-contrast microscopy. HUVEC migration was determined by wound healing method. The mRNA and protein expression of vascular endothelial growth factor A (VEGF-A) and basic fibroblast growth factor (bFGF) in both HUVEC and human colon adenocarcinoma cells (HT-29) was examined by reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immune sorbent assay (ELISA), respectively.

RESULTSPZH treatment significantly reduced the total number of blood vessels compared with the untreated control in the chicken embryos and resulted in a significant decrease in capillary tube formation and cell density of HUVECs (P<0.05). In addition, treatment with 0.25-1 mg/mL of PZH for 24 h, 48 h, and 72 h respectively reduced cell viability by 9%-52%, 24%-87% or 25%-87%, compared with the untreated control cells (P<0.05). Moreover, PZH treatment decreased the migration of HUVECs. Furthermore, PZH dose-dependently suppressed the expression of VEGF-A and bFGF on both mRNA and protein levels (P<0.05).

CONCLUSIONPZH could inhibit angiogenesis in vivo in CAM model and in vitro on HUVECs, suggesting that inhibiting tumor angiogenesis might be one of the mechanisms by which PZH treats cancer.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; HT29 Cells ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Neovascularization, Physiologic ; drug effects ; genetics ; RNA, Messenger ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism