Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10％ fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P＜0.05),and significant differences were found between different contrations of colchicine groups(all P＜0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P＜0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) ％,(21.04 ± 4.56) ％,(31.84 ±6.21)％and(35.32±5.56)％ in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P＜0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P＜0.05).The expression of the TGF-β in the cells was (97.60± 2.09) ％ in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) ％,presenting a significant difference between them (t =65.330,P ＜ 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion

Objective To study the recent clinical efficacy of compound matrine injections combined with three-dimensional conformal intensity modulated radiotherapy in the treatment of locally advanced pancreatic cancer.Methods 48 cases of untreated locally advanced pancreatic cancer were randomly divided into treatment group (compound matrine injections combined with three-dimensional conformal intensity modulated radiotherapy) and control group (three-dimensional conformal intensity modulated radiationtherapy),there were 24 cases in each group.Three-dimensional conformal intensity modulated radiation therapy used 95 ％ isodose coverage PTV.Using 6 MV X-ray irradiation,2 Gy/day,5 times/week,the total radiation dose was 50 Gy/25 times/5 weeks to complete.Treatment group:one week before treatment infusion of compound matrine injections 20 ml/time,l/d,21-day was one cycle.Control group:simple intensity modulated radiotherapy.Results After treatment,the clinical benefit response rate of the treatment group was 87.5 ％ (21/24),the control group was 70.83 ％ (17/24),the difference was statistically significant (P =0.045).Term efficacy rate of the treatment group was 79.17 ％ (19/24),the control group was 50.00 ％ (12/24),the difference was statistically significant (P =0.035).Adverse reactions were hematologic toxicity,leukopenia occurred in the treatment group was 16.67 ％ (4/24),but the control group was 45.83 ％ (11/24),the difference was statistically significant (P =0.029).Conclusion The effects of compound matrine injections combined with three-dimensional conformal intensity modulated radiotherapy for locally advanced pancreatic cancer are good,they can relieve pain,improve short-term efficacy,reduc side effects,and improve the quality of life.

5srRNA was isolated and purified from the reticulocytes of rabbit. It labeled with ~(125)I, then incubated with mouse myeloma cells (SP2/0). By autoradiography it was observed that ~(125)I-SsrRNA could pass into the nuclei of the cells. In a separate experiment, it showed that the incorporation rate of ~3H-TdR into nuclei of SP2/0 after incubation with 5srRNA was decreased as compared with that f control group, hence the result indicates that 5srRNA inhibits DNA synthesis of the SP2/0 cells and it seems to play a role in the regulation of gene expression through its hybridization with DNA sequences of the SP2/0 cells. Thus it is likely that 5srRNA might act as "erythroid denucleation factor".

The present study reported that 5srRNA from reticulocytes of rabbit could pass into the nucleus of mouse myeloma cells SP2/0,and in the mean time DNA synthesis and cells division were markedly suppressed. In a separate experiment, the rRNA was extracted from embryonic liver and erythrocytes of rabbit or rat and analysed by agarose electrophoresis method. The result indicated that the amount of 5srRNA in various period of the development of erythrocyte is changed along with denucleation process. Thus it is likely that 5srRNA of mammalian erythrocyte plays a role in reversing malignant phenotype of tumor cells and promoting denucleation of mammalian erythrocyte through inhibiting DNA synthesis.

Objective To assess the reliability and validity of the Chinese version of Multidimensional fatigue symptom inventory-short form(MFSI-SF).Methods The reliability and validity of the Chinese version MFSI-SF were assessed in a sample of 203 cancer patients.Statistical software was used to perform the analysis.Results The results showed moderate correlation between items and the total scale,the content validity index was 0.82,and exploratory factor analysis indicated five dimensions of the scale,the cumulative variance contribution was 56.65％.Confirmatory factor analysis showed moderate model fitting:x2/df=l.73,GFI=0.83,AGFI=0.79,NNFI=0.94,RMSEA=0.06,criterion validity was 0.585,and the Cronbach α of the total scale was 0.896.Conclusions The results demonstrated good convergent validity,it is suitable to evaluate fatigue status in Chinese cancer patients.

OBJECTIVERNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.

METHODSThe pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.

RESULTSSEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).

CONCLUSIONResults indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.

Animals ; Cleft Palate ; Gene Silencing ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; Oxidoreductases Acting on CH-CH Group Donors ; Palate ; growth & development ; RNA, Messenger

The IgG binding domain of Streptococcal Protein G which can selectively immobilizes the Fc regions of immunoglobulin G(IgG) is a kind of good material for oriented immobilization of antibodies in antibody microarrays.Here,genetically engineered three glutathione S-transferase(GST) fused proteins,bearing one,two and three B-Domains respectively(GST-GBx).The IgG-bindding ability of GST-GBx was investigated by ELISA.The data revealed that when the B-domain's quantity of GST-GBx is identical,the GST-GB3 is the most efficient protein among three GST-GBx protein both the capacity and sensibility of binding IgG.The GST-GB2 is the next one and GST-GB1 is the least one.Thus,the GST-GB3 has significantly predominance in comparison to GST-GB2 and GST-GB1.

Objective To investigate the incidence and influencing factors of healthcare-associated infection (HAI) in inpatients with nasopharyngeal carcinoma during radiotherapy period.Methods The occurrence of HAI among patients with nasopharyngeal carcinoma in a tertiary first-class cancer hospital in Jiangsu Province between July 2012 and June 2014 were analyzed retrospectively.Results A total of 1 396 patients were investigated,the incidence of HAI was 2.29%,case incidence of HAI was 2.44%.The most common infection site was oral mucosa (n =24, 70.59%),and most infection occurred 2-4 weeks after the start of the radiotherapy.A total of 38 pathogenic iso-lates were isolated,24 (63.16%)were gram-positive bacteria,12 (31 .58%)were gram-negative bacteria,and 2 (5.26%)were fungi.Incidences of HAI were high in patients >50 years old,with chemotherapy,length of hospital stay>60 days,and used at least 2 kinds of antimicrobial agents (all P <0.05).Conclusion Prevention and control of HAI in patients with nasopharyngeal carcinoma during radiotherapy period should be strengthened,especially for the elderly,patients with chemotherapy,long length of hospital stay,and extensive use of antimicrobial agents.

Objective To explore the influences of decorin ( DCN) gene down-regulating on biological behavior in human lung adenocarcinoma A 549 cell line.Methods Chemically synthesis in vitro targeting DCN -siRNA was transfected into human lung adenocarcinoma A 549 cells by LipofectamineTM 2000 .The gene expres-sion of DCN was detected using Real-Time PCR ;The protein expression of DCN was investigated using Western blot;Checkout DCN-siRNA effects on lung adenocarcinoma cancer cell proliferation was detected by CCK -8;The migration and invasion ability were determined by Transwell assay .Results DCN-siRNA was successfully transfected into human lung adenocarcinoma A 549 cells.Real-Time PCR results showed that DCN mRNA ex-pression level significantly decreased ,compared with untransfected group ,negative control group .Western blot re-sults showed that DCN-siRNA transfection inhibited DCN protein expression level;CCK-8 results showed that the proliferation was enhanced in the DCN -siRNA group as compared with untransfected group ,as well as the negative control group and empty vector group .Transwell assay results showed the invasion of A 549 cells in DCN-siRNA group was significantly enhanced as compared with untransfected group [(22.6 ±1.14) vs.(5.2 ± 0.84)].Wound-Healing assay results revealed that the cell repairing rate was markedly increased in DCN -siRNA group.A549 cells were close to complete repair after 48 hours.Apoptosis was not significant in DCN -siR-NA group as compared with untransfected group (P=0.214).Conclusion Down-regulation the expression of DCN gene by DCN-siRNA may enhance the ability of invasion ,migration and proliferation of A 549 cells without affecting apoptosis ,which provides a new evidence of function for advancing research of lung cancer .

Aim To establish ICR animal model with C6 glioma stem cells, to provide the ideal model for the further study of gli-oma stem cells in brain glioma model. Methods C6 glioma stem cell was cultured in vitro by suspension,and was identified with Nestin antibody. C6 stem cells of ICR mouse glioma model were used to investigate survival state and tumor volume in mice after the operation. HE staining and CD133 immunohistochemi-cal study were adopted to investigate the postoperative pathologi-cal changes in mice. Results The expression of Nestin was 96. 01% in C6 glioma stem cells, and Nestin was highly ex-pressed in the cultured C6 glioma stem cells. Mice were inocula-ted with tumor after loss of appetite, weight, behavior and slow, sluggish reaction. Tumor volume at day 21 after modeling was (9. 77 ± 6. 58) mm3 . After HE staining, the model showed the invasive growth, tumor cell shrinkage and derangement. Immu-nohistochemical CD133 staining revealed that tumor cytoplasm color was brown. Conclusion Glioma model can be established based on glioma stem cells into a high rate of tumor, the tumor cycle is short, which can be used as an ideal model for glioma.