OBJECTIVE:To prepare the curcumin solid lipid nanoparticles. METHODS:Microemulsion method was adopted to prepare curcumin solid lipid nanoparticles (SLN) and pseudo-ternary phase diagrams was conducted to optimize the preferable three-phase microemulsion factors. The optimal conditions were determined,and then hot microemulsion was dispersed in cold wa-ter to prepare SLN. After the preliminary screening of the factors (emulsifiers,lipid materials,amount of lipids,drug-to-lipid ra-tio,cold water phase temperature and the holding temperature of microemulsion)by single-factor test,with the index of encapsula-tion efficiency,the orthogonal test was conducted to optimize the formulation,followed by the verification test. RESULTS:The best microemulsion was produced at 65 ℃ with three phases consisting of stearic acid(oil phase),polysorbate 80(emulsifier)and ethanol (co-emulsifier) and the Km ratio of 1∶4. The optimal conditions for SLN were as follows as curcumin dosage of 50 mg, stearic acid of 0.5 g,cold water phase temperature of 2 ℃,holding temperature of microemulsion of 65 ℃. The encapsulation effi-ciency was 87.73% for solid lipid nanoparticles prepared,drug-loading rate was 7.72%,particle size was(156.9±2.2)nm,poly-dispersity coefficient was 0.480,average Zeta potential was -24.8 mV(RSD<2%,n=3). CONCLUSIONS:The microemulsion method to prepare SLN is simple and feasible.

Objective To explore the most suitable chemostatic condition and a method of cell-counting in reconstituted basement membrane invasion experiment in vitro. Method Reconstituted basement membrane invasion experiment was employed, and conditioned medium, fibronectin (FN), and a combination of conditioned medium and FN were respectively used as chemoattractants was placed in the lower compartment of the chamber. After the filters were stained with hematoxylin and eosin (HE), two kinds of cell-counting methods were employed. One was random cell-counting to count the cells in 5 fields infiltrated in the different areas of the filter membrane under 40?magnification, and the other was whole cell-counting of the infiltrating cells on the whole filter membrane with 4? magnification and image analysis system. Results The number of infiltrating cells in the presence of the chemoattractants was much larger than that of the control group. The number of infiltrating cells of the conditioned medium group was larger than that of FN group. The numbers of infiltrating cells in the combination of conditioned medium and FN groups were significantly larger than those of conditioned medium and FN alone groups. The accuracy of method of random cell-counting under 40? magnification was poorer than that of whole cell-counting with image analysis system. Conclusion Chemoattractants exert strong effect on the tumor cells invasion, and different chemoattractants show different chemotactic abilities. By using image analysis system to count all the infiltrative cells of the whole filter membrane is an objective and accurate cell-counting method.

Objective To investigate the effect of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment,and then to discuss if it's effect on endometrial receptivity through its controlling the expression of cell factors which related to endometrial receptivity.Methods Mice's endometrium tissue samples and uterine Natural Killer (UNK) cells were obtained from pregnant mice on gd 4.5 days and gd 12.5 days.After endometrial glandular epithelial cells,endometrial stromal cells and UNK cells were separated through primary culture,epithelial cells and endometrial stromal cells at rate of 1 ∶ 1 a certain concentration UNK were mixed together and highly purified-cultured.Different concentrations of Zidan decoction was acted on the culture solution.FBS,black and aspirin control group were setup for contrast.The expression of LIF and VEGF which influenced by Zidan decoction in the immunity environment was assayed by Realtime-PCR.Results The expression of LIF and VEGF was significantly increased in the immunity environment of uterine (in 10％ Zidan decoction group,10％ aspirin group and 10％ blank control group:LIF was 2.10±0.20,1.98±0.14,0.90±0.05 separately,and VEGF was 3.17±0.31,2.52±0.09,0.92±0.06respectively,P＜0.05).Conclusion Zidan decoction could improve endometrial receptivity by increasing the expression of cell factor which related to endometrial receptivity with the help ofUNK paracrine.

Objective To explore the effect of erlotinib in patients with non-small-cell lung cancer (NSCLC)in stage ⅢA before operation and the relation with the rate of resection,good operability and postoperative complications.Methods 31 patients with NSCLC in stage ⅢA in group A were treated with erlotinib before operation;34 patients with NSCLC in stage ⅢA in group B only were treated with operation.Results The condition of 64.5%patients were improved.The operability in group A was more than that in group B(P=0.008),the good operability in group A was more than that in group B(P=0.011),the postoperative complications do not have statistical significance (P=0.07).Conclusion The erlotinib can increase the rate of resection in patients with NSCLC in stage ⅢA,and increase good operability in patients with NSCLC in stage ⅢA,but not increase the postoperative complications.

Objective To compare the similarities and differences of the five detection methods used in the detection of fungi in vaginal secretions,and find the most sensitive、the most specific、the fastest、the most cost effective and the simplest method used in the detection of fungi in vaginal secretions.Methods A total of 442 patients were selected from the Department of Gynecology of Shenzhen OCT Hospital from May 2016 to August 2016.The vaginal secretion of 442 specimens was detected by using the methods of fungi culture、saline and KOH suspension method,Gram stain,Wright''s stain and Vaginitis Multi Test Kit.In these five methods,Fungi culture were using as gold standard to evaluate the specificity,sensitivity,negative predictive value,positive predictive value and accuracy of the other four methods.Results Using the fungus culture method to detect 442 cases of vaginal secretion,we found the positive rate of mycotic infection was 34.8%(154/442).Compared with the fungi culture method,the Specificity of saline and KOH suspension method was 97.9%,the sensitivity was 64.9%,the negative predictive value was 83.9%,the positive predictive value was 94.3% and the accuracy was 86.4%;the Specificity of Gram stain was 96.5%,the Sensitivity was 83.1%,the negative predictive value was 91.4%,the positive predictive value was 92.7% and the accuracy was 91.8%;the Specificity of Vaginitis Multi Test Kit was 84.7%,the Sensitivity was 46.8%,the negative predictive value was 74.8%,the positive predictive value was 62.0% and the accuracy was 71.5%;the Specificity of Wright''s stain was 96.9%,the Sensitivity was 78.6%,the negative predictive value was 89.4%,the positive predictive value was 93.1% and the accuracy was 90.5%.Conclusion Gram stain could be the most sensitive and specific method in the four methods,with highest accuracy,and the the fastest,the most cost effective and the simplest method for the detection of fungi in vaginal secretions.The accuracy of detecting fungi in vaginal secretions could be improved by the combination of Gram stain method in clinical work.

Objective To estimate the consistency between the diagnostic criteria for dermatomyositis (DM) and polymyositis (PM) developed by Bohan and Peter criteria (B/P criteria) and ENMC criteria.Methods The clinical,laboratory and pathological data from 86 patients who were initially diagnosed with idiopathic inflammatory myopathy were collected retrospectively.These patients were diagnosed according to B/P criteria and ENMC criteria,and the similarities and differences between these two criteria were compared.The data were analyzed with Mann Whitney U test and Kappa test by SPSS 13.0 software.Results Thirtyseven DM and 49 PM were diagnosed using B/P criteria.Forty-six DM and 14 PM were diagnosed using ENMC criteria,and 1 was diagnosed as eosinophilic myositis,9 were diagnosed as sporadic inclusion body myositis (sIBM),11 cases were diagnosed as limb-girdle muscular dystrophy type 2B,and the diagnosis of 5 patients could not be clarified.Agreement for DM between these two sets of criteria was very good by Kappa test (κ=0.79),but the corresponding between the two tests for PM was poor (κ=0.26).Conclusion Our study has demonstrated that B/P criteria may cause over-diagnosis and misdiagnosing for PM.ENMC criteria involves immunohistochemical pathology,stratified clinical and pathological exclusion criteria.The diagnostic accuracy of ENMC criteria is much improved.

Altitude ; Altitude Sickness ; Animals ; Hypoxia ; Iron ; Mice ; Spleen

Objective To continue to study if there is any other pathogenic gene expression related to Th1/Th2 abnormal differentiation,based on the author′s previous results,which have shown that Th1/Th2 unbalance is due to the cytokines and cytokine receptors of differentiation.Methods TaqMan Real time PCR was used to detect the gene expression of Th1/Th2 control in recent onset systemic lupus erythematosus (SLE) patients ( n =38).The genes include I?B,IRF 1,STAT4,GATA3,IL 4R and the others such as CCR1,CCR2,CCR4,CCR5,caepase 1 and CD38,which participate in inflammation,cell apoptosis and so on.Rheumatoid arthritis (RA) patients ( n =50) and normal people ( n =28) were control groups.Results ① Resent onset SLE patients comparing to normal people:STAT4 expression in IL 2/IL 12R ? 2/STAT4 access which induced Th1 differentiation increased significantly ( P 0 05) ;GATA3 expression which induced Th2 differentiation in IL 4/IL 4R/GATA3 access decreased significantly ( P

Objective To study the pathophysiology of ankylosing spondylitis (AS)-related osteoporosis by investigating the relation between bone mineral density (BMD) and bone turnover markers.Methods Fortysix AS patients were included in this study,including 35 male and 11 female patients.Twenty-five gender and age matched healthy subjects were served as the healthy control group.Informed consents were obtained from all subjects.BMD of lumbar spine (L2-4),fermoral neck and radius were tested using dual-energy X-ray absorption method.Data about BASDAi,ESR,Ig was collected.Bone formation markers including procollagen type Ⅰ N-terminal peptide (PINP) and osteocalcin (OC) were included for analysis,the formal was measured by radioimmunoassay and the later was measured by immunoradiometric assay and bone resorption marker.β-Crosslaps was measured by electrochemiluminescence immunoassay.Independent-samples t test was used to compare normal distributed data between the two groups.Analysis of variance was used to compare the data between the three groups.For those data which were not normally distributed,Rank sum test was performed.Correlations were determined by Pearson or Spearman's ranking.Results ① 48％ of AS patients had bone loss,while the percentage in the healthy control group was 20％ (x2=5.32,P=0.021).BMD of lumbar spine,fermoral neck and radius of the AS patients were lower than the controls (t=-3.73,-3.68,-5.24; P＜0.05).② Bone density (BMD) of lumbar spine in patients at the late stage was higher than patients at early disease stage (t=2.26,P=0.034),however,the BMD in the femoral neck was not.③ The procollagen (PINP),OC and β-CrossLaps were not significantly different in patients at different stage of diseases (P＞0.05).④BMD of lumbar spine was negatively correlated with the β-CrossLaps and ESR(r=-0.325,P=0.047; r=-0.314,P=0.046),The BMD in fermoral neck was negatively correlated with β-CrossLaps and OC (r=-0.387,P=0.024; r=-0.371,P=0.034),but they were not correlated with disease duration and BADSAI.Conclusion Osteoporosis is common in AS.Bone turnover markers including bone formation and bone resorption are increased in AS.β-CrossLaps is likely to predict bone loss in AS.

Objective To investigate the biological effects of exosomes secreted by KYSE410 cells on migration and invasion of KYSE410,KYSE510,YES2 cells and the possible mechanisms underlying the phenotype change.Methods The exosomes were isolated from the conditional supernatant of esophageal cancer cell line KYSE410 by ultracentrifugation.The morphology of exosomes was observed by transmission electron microscopy (TEM).Western blotting was used to detect the protein markers of exosomes.The uptaken of fluorescence-labeled KYSE410 exosomes by KYSE410,KYSE510 and YES2 was also recorded under confocal microscopy.Migration and invasion ability of the three esophageal carcinoma cell lines and the effects of exosomes from KYSE410 on migration and invasion of KYSE410,KYSE510 and YES2 cells were analyzed by Transwell chamber,respectively.The alteration of Wnt/β-catenin and PI3K/Akt pathway-related proteins were detected by Western blotting.Results The membrane structure of KYSE410 derived exosomes could be observed with its diameter ranged between 30-100nm.The invasion and migration ability of three esophageal cancer cells are KYSE410＞ KYSE510＞ YES2.KYSE410 exosomes promoted the migration and invasion of KYSE410,KYSE510 and YES2 cells.Conclusions Concentrated exosomes derived from the highly migratory and invasive esophageal cancer cell line KYSE410 promoted the migration and invasion potentials of itself and esophageal cancer cell lines KYSE510 and YES2,which possibly exerted the effects by activating Wnt/β-catenin and PI3K/Akt signaling pathways.