AIM: To investigate the mechanism of basil polysaccharide in inhibiting the invasion and metastasis ability of ovarian cancer,and to evaluate the potency of hypoxia tumour microenvironment in affecting the action of basil polysaccharide.METHODS: The human ovarian cancer cell line SKOV3 was cultured with basil polysaccharide under normoxia(21% O2,5% CO2) and hypoxia(1% O2,5% CO2,94% N2),respectively.Morphology of this cell line in different groups under different oxygen concentrations was observed under inverted phase contrast microscope.The cell invasiveness was tested by transwell method and the activities of matrix metalloproteinase-2(MMP-2) among each group were analyzed by gelatin zymography.The expressions of OPN mRNA and protein levels were detected by RT-PCR and immunohistochemistry,respectively.RESULTS: Compared to the control group,basil polysaccharide decreased the invasiveness and migrating ability and inhibited the secretion of MMP-2 in SKOV3 cells.Basil polysaccharide also reduced the expression level of osteopontin(transcriptional level and protein level) of the SKOV3 cells in both normoxic and hypoxic conditions.Moreover,these functions of basil polysaccharide were more obviously in hypoxia.CONCLUSION: The migrating ability of the SKOV3 cells is enhanced under hypoxic condition.Basil polysaccharide may inhibit the invasiveness and migrating ability of SKOV3 cells through reducing osteopontin expression and MMP-2 secretion.

Objective To observe the combined poisonous effects of fluoride and aluminum on sex hormone of male rats.Methods Sixteen weaned SD healthy male rats aged two week were selected and divided into control group,aluminum group,fluoride group,fluorine-aluminum group,four rats in each group.All rats in the experimental groups were fed with corn collected from the prevailng areas containing different fluorine contents respectively for 90 days.Serum testosterone(T)and estradiol(E2)were detected.Results Compared separatelv with the control group[(3.317±0.635)μg/L],serum T level of fluorine-aluminum group[(15.994±6.558)μg/L]was higher(P＜0.05),but aluminum[(8.134±3.134)μg/L]and fluorine[(1.868±0.367)μg/L]groups had no significant differences(P＞0.05).Compared separately with the control group[(0.319±0.072)nmol/L],E2 level of the fluorine group[(0.172±0.030)nmol/L]being lower(P＜0.05),and it was not significant differences(P＞0.05)in the control group when compared with aluminum group[(0.282±0.012)nmol/L],and fluorine-aluminum group[(0.265±0.047)nmol/L].Fluorine and aluminum interacted with each other(F=9.82,P＜0.05).Conclusion The combined poisonous effects of fluorine and aluminum may influence sex hormone levels of male rats.

OBJECTIVETo evaluate the clinical efficiency of selective cyclo-oxygenase-2 (COX-2) inhibitor compared to traditional nonselective NSAIDs for the prevention of heterotopic ossification (HO) after total hip arthroplasty (THA).

METHODSBy searching Medline, Embase, CENTRAL (Cochrane Central Register of Controlled Trials) and Science Citation Index et al, only randomised controlled studies of selective COX-2 inhibitors VS nonselective COX-1 and COX-2 inhibitors for the prevention of HO after THA were included. The quality assessment of included studies was evaluated according to the standard of the Cochrane Collaboration, and the data were analysised by statistic software Stata 10.0. The HO incidence of both groups in different degrees was compared.

RESULTSFour eligible randomised controlled trials of totally 808 patients were included. Meta-analysis results showed that no statistically significant difference was found in overall incidence of HO (RR = 1.08, 95% CI: 0.71-1.64,P = 0.73), incidence of moderate severe HO (Brooker II and III) (RR = 0.83, 95% CI: 0.48-1.42, P = 0.49) and any grade of Brooker classification between two groups. In all included studies, 16 patients receiving nonselective COX inhibitor (4.4%) discontinued treatment because of gastrointestinal toxicity,whereas 10 patients in the selective COX-2 inhibitor group (2.7%) discontinued for gastrointestinal side effects.

CONCLUSIONThe selective COX-2 inhibitors are as equally effective as nonselective NSAIDs for the prevention of HO after THA. Considering the side effects of nonselective NSAIDs, selective COX-2 inhibitors were recommend for the prevention of HO after THA.

Anti-Inflammatory Agents, Non-Steroidal ; adverse effects ; therapeutic use ; Arthroplasty, Replacement, Hip ; adverse effects ; Cyclooxygenase 2 Inhibitors ; adverse effects ; therapeutic use ; Cyclooxygenase Inhibitors ; adverse effects ; therapeutic use ; Humans ; Ossification, Heterotopic ; prevention & control ; Randomized Controlled Trials as Topic

OBJECTIVETo study the kinetogenic effects of serum containing Semen Arecae, Dandelion, Semen raphani and Atractylodes macrocephala on the colonic smooth muscle cells of rats.

METHODSSerum containing Chinese materia medicas was made according to standard methods. Smooth muscle cells were isolated from the muscle layers of Wistar rat's colon, referred to modified Bitar's method. The contractile response of colonic smooth muscle cells to serum containing Chinese materia medicas (10%, 50%, 100% concentration) and other medicines (blank and 1 x 10(-3) mol/L acetylcholine) were separately observed. The contractility was presented by the decrease of the cell length between the drug groups and the control.

RESULTSSerum containing each Chinese materia medica can make dose-dependent contraction at different concentrations (P < 0.05), but the strongest effect of each serum had no significant difference (P > 0.05).

CONCLUSIONSerum containing Semen Arecae, Dandelion, Semen raphani and Atractylodes macrocephala can make notable contraction on colonic smooth muscle cells in rats.

Animals ; Cells, Cultured ; Colon ; cytology ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Muscle Contraction ; drug effects ; Muscle, Smooth ; cytology ; Myocytes, Smooth Muscle ; drug effects ; Rats

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.

Objective To construct a recombinant bacillus Calmette-Guérin(BCG) vaccines based on different tandem repeats of MUC1 and GM-CSF, rBCG-MVNTR1/4/8-CSF, and to observe the ability of three recombinant BCG vaccines in the inhibition of breast cancer. Methods After MUC1 variable-number tandem repeats (MVNTR1/4/8) were cloned in a stepwise manner, the E. coli-Mycobacteria shuttle expression vector pDE22-MVNTR1/4/8-CSF were constructed by fusing MVNTR1/4/8 and GM-CSF, and then used to transform competent BCG by electroporation after identification by restriction endonuclease digestion analysis and DNA sequencing. A novel breast cancer vaccines, rBCG-MVNTR1/4/8-CSF was constructed. The expression of fused MVNTR1/4/8-CSF protiens was analyzed by SDS-PAGE and Western blot. The ability of rBCG vaccines inhibiting the growth of breast cancer was observed in hu-PBL-SCID mice. The specific T cell responses in mice were assessed by immunohistochemistry. Results The expression of recombinant MVNTR1/4/8-CSF fusion proteins were detected by SDS-PAGE and Western Blot in rBCG-MVNTR1/4/8-CSF vaccines, respectively. Tumor incidence in mice prophylactic immunized with rBCG-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) significantly decreased compared with PBS and BCG-pDE22 control ( 100% ) at 42 days after tumor implantation ( P ＜ 0. 05 ). MCF-7 tumor growth inhibition in rBCG- MVNTR4/8-CSF-immunized mice was more significant than that in controls ( P ＜0. 01 ).The inhibition effect of three rBCG vaccines on breast rumor growth appeared to rise with increase of numbers of the tandem repeats of MUC1. Survival rate was 75% of mice in the rBCG-MVNTR4-CSF-treated group and 87. 5% of mice in the rBCG-MVNTR8-CSF-treated groups at 70 days after tumor implantation; however,survival rate was only 12. 5% in control group( P ＜0. 05). The CD4-positive and CD8-positive lymphocytes were detected only in rBCG-MVNTR4/8-CSF-immunized mice. Conclusions rBCG-MVNTR4/8-CSF immunization inhibits breast cancer growth in mice.

Objective To compare the results of susceptibility testing by European Committee on Antimicrobial Susceptibility Testing (EUCAST)and Clinical and Laboratory Standards Institute (CLSI)broth microdilution methods against Aspergillus isolates.Methods The susceptibilities of 116 Aspergillus isolates were determined for amphotericin B, voriconazole, itraconazole,caspofungin and micafungin according to EUCAST (E.DEF 9.1 )and CLSI (M38-A2)methods.The essential agreement (EA),categorical agreement (CA),very major errors (VME)and major errors (ME)of the two methods were compared.Results The EA was 96.3%-100% between the two methods.The CA ,ME,and VME were 98.8%,0-1.2% and 0 respectively for the susceptibility of Aspergillus fumigatus to voriconazole.The CA,ME and VME was 1 00%,0 and 0 respectively for the susceptibility of Aspergillus fumigatus and Aspergillus niger to amphotericin B,or the susceptibility of Aspergillus fumigatus and Aspergillus flavus to itraconazole.Conclusions The results of susceptibility testing by EUCAST and CLSI broth microdilution methods are well consistent against Aspergillus isolates.

Objective To identify the efficacy of granulocyte-macrophage colony-stimulating factor (GM-CSF) gargle in patients who had oral mucositis after allogeneic hematopoietic stem cell transplantation. Methods A total of 134 patients were enrolled in this study from 2014 to 2015. They were randomly divided into two groups:the experimental group (n=65) and the control group (n=69). Both groups received preventive measures for oral mucositis. But once oral mucositis occurred, the control group continued with the routine nursing measure, while the experimental group added GM-CSF gargle based on previous routine nursing measure. The effective rate and healing time were compared between two groups. Results The effective rate of the experimental group (81.54%, 53/65) was significantly higher than that of the control group (24.64%,17/69) (χ2=43.434, P=0.000). The median healing time in the experimental group was 4.5 days, shorter than 9.0 days in the control group (Z=-5.379, P< 0.01). Conclusions GM-CSF gargle can control the oral mucositis after allogeneic hematopoietic stem cell transplantation.

Objective To investigate the nutritional status of vitamin D in preterm infants after birth and further explore its possible influencing factors, so as to guide clinical vitamin D therapy and to screen the preterm infants who are at high risk of vitamin D deficiency.Methods Retrospective analysis was conducted in the neonatal department of our hospital from April 21st, 2014 to February 5th, 2016.The serum 25(OH)D level in preterm infants were measured 2 weeks after birth.Data including gender, season of birth, time to initiation of breastfeeding were collected.According to the 25(OH)D levels[25(OH)D≤37.5 nmol/L, 37.5 nmol/L≤50.0 nmol/L, and 25(OH)D>50.0 nmol/L], all the preterm infants were divided into three groups: vitamin D deficiency, insufficiency, and sufficiency groups.The influencing factors of vitamin D in preterm infants were screened by using statistical method.Results The mean 25(OH)D level of 172 preterm infants was (43.1±16.7)nmol/L.In vitamin D deficient, insufficient, and sufficient groups, there were 68 (40％), 50 (29％) and 54(31％) cases of preterm babies, respectively.The mean values of 25(OH)D in these three groups were (27.8±16.7)nmol/L, (42.4±3.4)nmol/L, and (63.0±11.7)nmol/L, respectively.Only the season of birth had significant difference among three groups (P=0.013): 44.2％ of the preterm infants born in winter had vitamin D deficiency, which was higher than those in spring (41.7％), summer(33.3％), and autumn (38.1％);44.2％ of the preterm infants born in winter had vitamin D insufficiency, which was much higher than those in spring (30.6％), summer (25.1％), and autumn (19.0％);furthermore, only 11.6％ of the preterm infants born in the winter had vitamin D sufficiency, which was much lower than those in spring (27.8％), summer (41.2％), and autumn (42.9％) (OR=4.655, 95％ CI=1.716-12.627, P=0.003).Conclusion Vitamin D deficiency in preterm infants 2 weeks after birth is prevalent, and winter birth is a risk factor of vitamin D deficiency and insufficiency in preterm infants.

?AIM: To evaluate the effects on recovering of corneal wound and postoperative discomfort of different methods for primary pterygium.?METHODS: Forty-seven cases ( 60 eyes ) of primary pterygium were excised under microscope with limbal epithelial transplantation, with sharp dissection ( 24 cases, 30 eyes, Group A) and blunt dissection (23 cases, 30 eyes, Group B).All cases were followed up for 1d to 1mo.?RESULTS: The recovering of corneal wound was better in Group B on 1st day and 3rd day after surgery.Pain, photophobia and tears, foreign body sensation were more serious in group A on 1st day after surgery with a statistically significant difference (P=0.005,0.015,0.012). Pain, photophobia and tears, foreign body sensation were more serious in Group A on 3rd day after surgery with a statistically significant difference ( P=0.019,0.018, 0.015).There was no statistically significant difference on 1wk and 1mo after surgery (P>0.05).? CONCLUSION: Compared with sharp dissection, primary pterygium excised with blunt dissection can significantly improve recovering of corneal wound and postoperative discomfort.