Objective:To optimize the extraction technology for Sanren Tongbian mixture. Methods:The water amount,decoction time and decoction frequency as the evaluation factors,and the content of amygdalin and the paste-forming rate as the evaluation indi-ces,the extraction process of Sanren Tongbian mixture was optimized by L9 (34 ) orthogonal design. Results: The optimal extraction conditions were as follows:adding 8-fold amount of water, decocting twice with 1. 5h for each time. Conclusion: The study provides scientific basis for the research of the extraction technology of Sanren Tongbian mixture, and the further experiments proved that the op-timized process is stable and feasible.

Objective To simultaneously determine the quantify liquiritin, glycyrrhetinic acid and glycyrrhizic acid inXianlong-Jiedu Yin by high performance capillary electrophoresis (HPCE).Methods HPCE method was performed on a fused silica capillary coates 75μm×85 cm (effective column length 78.5 cm) using 50 mmol/L borax solution-acetonitrile (95:5), with adjusted pH=8.50 as running buffer, lytical 22 kV, and electrokinetic injection 10 s, detected wave length of 254 nm, and temprature maintaining 25℃.Results The liquiritin, glycyrrhetinic acid and glycyrrhizic acid were completely separated within 30 minutes, showing good linear relationship in the range of 12-120, 15-150, 8-160μg/ml (r>0.999 3). The average recoveries were 100.70%, 98.51%, 98.40% withRSD 2.49%, 2.79%, 0.40%, respectively.Conclusions The HPCE was suitable for the determination of liquiritin, glycyrrhetinic acid and glycyrrhizic acid inXianLong-JieDu Yin, which can be used for the quality control.

Objective To systematically evaluate the effectiveness and safety of axillary lymph node dissection for breast cancer patients with negative sentinel node biopsy.Methods Literatures in CNKI,PubMed,EMBASE and CBMwere searched from their establishment to December 1,2013.Accord-ing to the inclusion and exclusion criteria,trials of axillary lymph node dissection and sentinel node biopsy for breast cancer were strictly screened and extracted for quality assessment and result analysis.Meta-anal-ysis was conducted by using Revman 5.1 software.Results A total of 10 studies involving 7731 patients were eligible for the final analysis.Because of the large differences in research type,measurement indica-tor,follow-up period and statistical index,subgroup analysis was adopted.Meta-analysis was applied for homogeneous researches and the remaining studies were analyzed with qualitative descriptive analysis.The results showed no significant differences in disease-free survival,overall survival,local recurrence rate,and distant metastasis rate in different follow-up periods.Conclusion For breast cancer patients with single invasive lesion,axillary lymph node dissection is not necessary.More high-quality random control trials and long-term follow-up are required to confirm the conclusions of this systematic review.

Objective To understand the awareness degree and the dietary intake of acrylamide (AA) in food among preventive medicine professional undergraduate students in a university of Xianyang city .Methods 28 kinds of fried and baked food frequently eaten by undergraduates were selected as the respondents .The food-frequency method was adopted to perform the questionnaire in-vestigation on 248 undergraduates sampled from grade 1-5 of preventive medicine professional undergraduate students by the clus-ter sampling .Results More than 38 .71% of the investigated students had never known about the acrylamide ,32 .26% of the inves-tigated students had heard but did not know what it was ,11 .26% students knew about its hazard .The AA dietary intake was about 31 .57 μg/d per person ,and there was no statistical difference in the AA dietary intake between different sexes and grades .Conclu-sion The awareness degree of AA among investigated preventive medicine professional undergraduates is relatively low .Therefore it is necessary to strengthen the publicity and education of the AA-related food safety knowledge .

Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

[ ABSTRACT] AIM:To study the expression profiles and the role of Ca 2+/calmodulin-dependent kinase II delta ( CaMKIIδ) during osteoclast differentiation .METHODS:Mouse RAW264.7 cells were induced by receptor activator of nuclear factor κB ligand ( RANKL) at 50μg/L for osteoclastogenesis .Tartrate-resistant acid phosphatase ( TRAP) staining and bone resorption lacunae examination were performed to verify osteoclast formation .The expression of CaMKIIδat mR-NA and protein levels was also determined by immunofluorescent cytochemistry , RT-qPCR and Western blot at days 0, 1, 3 and 5.RESULTS:TRAP positive multinuclear cells with bone resorption function were formed after 5 d of induction. The mRNA levels of CaMKIIδdetected by RT-qPCR were 1.028 ±0.041, 2.478 ±0.087, 10.524 ±1.284 and 42.914 ± 2.667 at days 0, 1, 3 and 5, respectively, while the protein levels of CaMKIIδ detected by Western blot were 0.762, 0.963, 1.802 and 3.136, respectively.The changes of protein level were also verified by immunofluorescence cytochemis -try, in which the fluorescence intensity increased in a time-dependent manner (P<0.05).CONCLUSION:The expres-sion of CaMKIIδincreases with the differentiation of osteoclasts .CaMKIIδmay play a key role in the osteoclastogenesis .

The effects of fucoidan on the lipidomics profiling of juvenile yellow catfish (pelteobagrus fulvidraco) during different feeding time (week 1, week 2, week 3 and week 8) were investigated by ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-Q-TOF-MS) combined with multivariate data analysis, e.g.principal component analysis (PCA) and partial least squares-discriminate analysis (PLS-DA).Based on VIP and p values, 11 lipid biomarkers were screened including lysophosphatidylcholine (Lyso-PC) 16∶0, phosphatidylcholine (PC) 22∶6/16∶0, phosphatidylethanolamine (PE) 22∶6/16∶0, phosphatidylinositol (PI) 18∶0/22∶6, diacylglycerol (DAG) 16∶0/16∶0,DAG 16∶0/18∶1, DAG 18∶1/18∶1, triglycerides (TAG) 20∶5/16∶0/18∶3, TAG 20∶4/16∶1/18∶2, TAG 16∶0/18∶2/20∶5 and TAG 18∶2/14∶0/18∶1.It was found that Lyso-PC, PC, PE and PI had the largest levels at the eighth week;the levels of DAG were decreased at the second and fourth weeks, and increased at the eighth week.While TAG was different: the content of TAG 18∶2/14∶0/18∶1, TAG 20∶5/16∶0/18∶3 and TAG 20∶4/16∶1/18∶2 increased in response to fucoidan, but TAG 16∶0/18∶2/20∶5 decreased little.Therefore, for juvenile yellow catfish, fucoidan can affect its lipid metabolism, which provides a theoretical basis for investigation of the influence of fucoidan on the response mechanism of lipid metabolism of juvenile yellow catfish.

Objective To study the effect of zoledronate (ZOL) on Ca2+/calmodulin-dependent kinase Ⅱ δ (CaMK Ⅱ δ) and down-stream gene expressions during osteoclast differentiation.Methods Mouse osteoclast precursors RAW264.7 cells were divided into the control group and ZOL group.The cells in both groups were induced with 50μg/L receptor activator of nuclear factor kappa B ligand (RANKL) and were harvested on 5 d,while the cells in ZOL group were also simultaneously treated with 1 × 10-6 mol/L ZOL for 2 d.Five days later,the cells were harvested and examined osteoclastogenesis,as well as gene expressions of CaMK Ⅱ δ,nuclear factor of activated T-cells cytoplasmic 1 (NFATc1),tartrate-resistant acid phosphatase (TRAP) and cell-sarcoma receptor coactivator (c-Src).Results The number of TRAP positive multinuclear osteoclasts,number and size of dentin absorption lacunae and area in the ZOL group were (20.0±3.2),(18.0±4.2) and (6 335.3± 1 043.2)μm2 respectively,which were significantly lower than (36.0 ± 8.4),(37.2 ± 5.0) and (11 636.2 ± 3 661.1) μm2 in the control group and decreased by 44.4 ％,51.6 ％ and 45.6 ％ respectively (P＜0.01).ZOL also significantly inhibited the gene expressions of CaMK Ⅱ δ,NFATc1,TRAP and c-Src,and the mRNA levels of these genes were decreased by 44.1％,49.0％,53.8％ and 49.6％ respectively,the protein level were decreased by 43.5 ％,32.2 ％,45.5 ％ and 48.0 ％ respectively.The immunofluorescent cytochemistry detection results showed the fluorescence intensity of CaMK Ⅱ δ,NFATc1,TRAP and c-Srcin in the ZOL group was significantly weakened when compared with the control group.Conclusion ZOL could significantly inhibit the osteoclast formation and bone absorption function,and down-regulates gene expressions of CaMK Ⅱ δ,NFATc1,TRAP and c-Src in osteoclast differentiation.

Objective To observe the effect of sodium butyrate on the invasion and migration of human salivary gland ade‐noid cystic carcinoma cell line ACC‐2 in vitro and to investigate the underlying mechanisms .Methods The cultured ACC‐2 cells were treated with different concentrations of sodium butyrate for 24 h ,and detected the viability rate of the cells by methyl thiazolyl tetrazolium(MTT) assay ;the drug′s influence on invasion and migration on ACC‐2 were detected by Transwell experiment ,while the protein and mRNA expression of HMGB1 and TLR4 explored by Western‐blot and RT‐PCR assay ;the relationship between TLR4 expression and HMGB1 was investigated .Results Compared with control group ,0 .625 ,1 .250 ,2 .500 ,5 .000 ,10 .000 mmol/L groups of sodium butyrate inhibited the proliferation of ACC‐2 cells(P<0 .05);the influence of sodium butyrate on the in‐vasion of ACC‐2 cells of all groups had no significant difference(P>0 .05);only 2 .500 ,5 .000 and 10 .000 mmol/L groups inhibited ACC‐2 cells migration and down‐regulated the protein and mRNA of HMGB1 and TLR4(P<0 .05) .Correlation analysis showed a positive correlation between the TLR4 protein and HMGB1 protein(r=0 .810 ,P<0 .05) .Conclusion Sodium butyrate could inhib‐it ACC‐2 cells proliferation and high concentration gropes inhibit ACC‐2 cells migration ,while reducing HMGB1 and TLR4 mRNA and protein expression ,suggesting that NaB might inhibite ACC‐2 cells migration through down‐regulated the mRNA and protein expression .

Various teaching methods were used in occupational health and occupational medicine teaching,including problem based learning,multimedia teaching,bilingual teaching,case based learning and practice teaching methods when being confronted with new situation of occupational health and occupational safety.These methods are mean to encourage students' enthusiasm,cultivate students' comprehensive ability and enhance their sense of social responsibility and mission.Results showed that these methods improved the quality of teaching and achieved good teaching results.