To investigate the inhibitory effects of siRNA expression vector on proto-oncogene Pokemon expression in SW480 cells and to provide experimental basis for further research about the biological function of Pokemon.siRNA expression vectors were constructed to express a short hairpin RNA against Pokemon.The recombinants were transfected into SW480 cells with liposome.Cellular morphology and transfection efficiency were observed.The expression of Pokemon were checked by real-time fluorescence quantitative PCR and Western blot.MTT assay was used to detect the effect of siRNA on the growth of SW480 cells and Fluorescence Activated Cell Sorter was applied to study cell apoptosis of SW480 cells.The transfection efficiency was about 36% and the cellular morphology changed greatly at 48h after transfection.siRNA expression vectors could specifically reduce the expression of Pokemon mRNA and protein in SW480 cells.Compared with negative control,the inhibition ratio of Pokemon mRNA expression was 34.2% and 67.7%,in 24th hour and 48th hour,the inhibition ratio of Pokemon protein was 48.3% and 73.6%,in 48th and 72th hour,respectively.Transfection with Pokemon siRNA expression vectors could inhibit cell growth and induce cell apoptosis of SW480 cells.siRNA expression vectors were successfully established that could effectively inhibit the expression of Pokemon in SW480 cells.Pokemon gene silencing could decrease growth and increase apoptosis of SW480 cells.

Natural medicines play an important role in keeping the healthy of human beings from past till now. Recently, the importance of natural medicines in research and development of new drugs has been attracted the attention of scientists. The research and development of anti-aging drugs from natural products have captured more and more attention. In this paper, the historical story on the discovery and study of the famous natural anti-aging drug-ginkgolides is summarized in memory of the great discoveries and in honor of the scientists so as to provide the reference for future related research.

Objective To construct adenovirus vector containing firefly luciferase reporter gene (AdLuc) and infect bone marrow mesenchymal stem cells (BMSC),then to take bioluminescence imaging in vitro and in vivo for identification.Methods The luciferase gene was amplified with PCR from psiCHECK-2 plasmid and cloned into the adenoviral shuttle vector (pShuttle-CMV).It was confirmed by Nhe Ⅰ/Xba Ⅰ digestion and sequencing.PShuttle-CMV-Luc and backbone vector (pAdeno) were homologous recombined.Then the recombinant plasmid was packaged in HEK293 cells and the virus titer was detected.The BMSC were infected by the recombinant adenovirus.The bioluminescence imaging in vitro was performed to determine the best multiplicity of infection (MOI),and the relationship between bioluminescence intensity and MOI was analyzed by curve fitting regression analysis.Viability was evaluated via Trypan blue staining.The transfected BMSC (l× 106) were implanted into the muscles of forelimb of SD rats,and then tracked by bioluminescence imaging in vivo.Cell viability was compared using two-way repeated measures analysis of variance between groups.Results Enzyme digestion and sequence analysis indicated that Ad-Luc was successfully constructed.The virus titer was 1 × 1010 plaque forming unit (PFU)/ml.The bioluminescence detection in vitro showed that Ad-Luc could infect BMSC high efficiently to express luciferase and the best MOI was 50.The bioluminescence intensity enhanced with increase of MOI (R2 =0.98).No statistically significant difference was found in cell viability between transfected and untransfected BMSC at 1,3,5,7 d.The cell survival rates were (92.5±2.3)％ vs (94.1±1.8)％,(91.4±0.9)％ vs (92.7±2.0)％,(92.1±1.6)％ vs (93.3± 2.4) ％,(91.9 ± 1.5) ％ vs (93.0 ± 3.1) ％,respectively (F =4.38,P ＞ 0.05).The bioluminescence imaging in vivo showed that BMSC survived 1,3,7 d after implantation.However,bioluminescence signal decreased gradually over time.Conclusion It is feasible to apply the optical reporter gene imaging for tracing transplanted stem cells in vitro and in vivo due to the effective transformation of luciferase reporter gene into BMSC by adenovirus vector.

The rate of complete surgical resection is still low in hilar cholangiocarcinoma,which greatly affects the curative effect.Radiotherapy,one kind of treatment for tumors,has not been widely adopted in the past years.In recent years,with the development of radiotherapy technology,research of treating hilar cholangiocarcinoma through radiotherapy has become a spot.Many studies have shown that radiotherapy,as an adjuvant therapy for surgery and non-surgical treatment,can be the major means for treatment,and it could bring benefit for patients' survival extension and improvement of life quality.

Objective To analyze the expression of phosphorylated protein kinase B(pAKT)and PTEN protein in ovarian epithelial cancer and to investigate the correlations between their expression and prognosis of ovarian epithelial cancers.Methods Expression of pAKT and PTEN in 12 normal ovarian tissues,20 benign tumors,12 borderline tumors and 80 cases of ovarian epithelial cancers were detected by immunohistochemical method,and their correlations were analyzed.Results The positive expression of pAKT in normal ovarian and benign tumor tissues were significantly lower than that in ovarian epithelial cancers(8%,10% vs 55%;P

The new drugs and the novel theories developed from natural products chemistry study have made important contribution for scientific progress. In this paper, the success achieved in natural product research was briefly reviewed, which would broaden the outlook of young researchers and arouse their interest in this area.

Quinine is a very famous natural medicine that has saved millions of lives, even being believed to affect the development process of human beings. The study of quinine has left a very important record in the history of science. In this paper, the historical stories on the discovery and study of quinine are summarized in memory of the great discoveries and in honor of the scientists. It also aims at providing the references for future related research. This article is one of the series of historical stories on natural product chemistry written by the authors.

Objective To analyze if there are membrane estrogen receptors(ER)in endometrial carcinoma and if there is some relationship between membrane ER and nuclear ER.Methods The cell membrane and total cell ER?and ER?expressions of high and moderate differentiation endometrial carcinoma cells(Ishikawa and HEC-1A cells)were analyzed.Intermittent immunofluorescence dyeing and fluorescent microscopy were carried out with the ceils treated with polyformaldehyde and Triton X-100. Intermittent immunofluorescence dyeing and flow cytometry were carried out with the live cells and the cells treated with Triton X-100 respectively.Results There were fluorescences on the membrane of the Ishikawa and HEC-1A cells which were treated with polyformaldehyde.When the cells were treated with Triton X- 100,the fluorescences were also seen inside the cells.The fluorescence intensity of ER?and ER?in Ishikawa cell membrane(1.09?0.21,1.27?0.33)was stronger than the control,but there were no significant differences(P＞0.05).When treated with Triton X-100,the total cell fluorescence intensity of ER?and ER?in Ishikawa cell(4.21?0.34,4.69?1.96)was stronger than the membrane(P＜0.05). The ER?and ER?fluorescence intensity of HEC-1A cell membrane(1.58?0.13,1.49?0.04)were stronger than the control(P＜0.05).The fluorescence intensity of ER?and ER?of the HEC-1A cell (2.34?0.33,2.52?0.15)was stronger than the membrane also(P＜0.05).The membrane ER?fluorescence intensity of Ishikawa was lower than HEC-1A(P=0.028).But the total cell ER?fluorescence intensity of Ishikawa was higher than HEC-1A(P=0.002).Conclusions There are membrane ER on endometrial carcinoma cells Ishikawa and HEC-1A.The membrane ER must have some similarity to the nuclear receptor.There is no direct correlation between the quantity of the membrane ER and nuclear ER.

ObjectiveTo identify the framework for assessing health related quality of life(HRQOL) of Kaschin-Beck disease(KBD),in order to reflect the impact of KBD on quality of life in patients with the disease.MethodsQualitative descriptive research was adopted.Semi-open ended questions were developed by using the World Health Organization(WHO) definitions of health and quality of life.Group interview and face to face interviews were conducted on 48 patients with KBD and 29 health care experts on KBD in Linyou and Yongshou counties,Shaanxi province,which were higher prevalence areas of KBD.Content template analysis was conducted and the template was based on the WHOQOL-100's framework.ResultsThe framework of HRQOL for KBD included four domains:physical activity,familial/social support,economic and psychological state.There were also eleven facets which were:pain and discomfort,physical function and activity limitation,diet and sleeping,social relationship,concerns of family responsibilities,social support,economic,housing and the surrounding environment,appearance concerns,mental health,and general state of health.The total entries were 69.ConclusionsThe framework for assessing HRQOL of KBD is established.The framework highlights the impact of KBD on the patients' quality of life with higher specificity.

Objective To observe the effect of low arsenic sub-chronic exposure on blood routine test index in rabbits to pave a way for screening early injury of the low arsenic exposure. Methods Twelve healthy male rabbits were randomly divided into four groups. They were administrated with As at the concentration of 0(control), 10, 50 and 250 μg/L in the drinking water. Blood samples were collected from the vein of the ear edge in 8 weeks, and blood test routine including leukocyte (WBC), lymphocyte (LYM), lymphocyte percentage (LYM%), neutrophil (GRA), neutrophil percentage (GRA%), monocyte (MON), monocyte percentage (MON%), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin content (MCH), mean corpuscular hemoglobin concentration(MCHC), RDW, platelets(PLT), mean platelet volume(MPV), platelet hematocrit(PCT) and platelet distribution width (PDW), were detected by the ABX-60 hemocyte analyzer. Results Compared with the control group, the WBC, GRA and GRA% increased in 0, 50 and 250 μg/L groups, but there was no significance(P > 0.05). PLT and MPV had a statistical significance in 4 groups(F = 4.07,4.20, all P < 0.05). Compared with the control group[(292.00±16.97)×109/L, (7.10±0.99)fL], PLT decreased in the 250 μg/L group [(221.33±22.50)×109/L] and MPV decreased in the 50μg/L group [(5.57±0.46)fL] significantly (P < 0.05). The other index didn't change obviously. Conclusions Sub-chronic low level arsenic exposure may lead to the change in the blood system. The blood routine test may be considered for early injury of the arsenic poisoning.