Objective To explore the influence of salvianolic acid B(SalB) to blood brain‐barrier(BBB) in vitro oxygen/glu‐cose deprivation and reintroduction .Methods We used the human brain microvascular endothelial cells and C6 cells to build BBB model that in vitro ,and the models was divided into normal groups ,experiment groups ,treatment groups .The Earle′s liquid and the condition of 93% N2 ,2% O2 ,5% CO2 replaced the normal culture environment to simulate environment of oxygen /glucose depriva‐tion and reintroduction .The changes of BBB permeability was detected by the horseradish peroxidase permeability test .The tight junctions(TJs) was detected by immunofluorescence and electron microscope .Results The permeability of treatment group was significantly lower than the experiment group (P < 0 .05) ;The TJs of normal group was continuous and density ;the experiment group′s TJs was interruption and osteoporosis ;and the observation of treatment group′s TJs was between of the two group above . Conclusion The SalB can directly protect the BBB during oxygen /glucose deprivation and reintroduction ,and decrease the permea‐bility .The investigation also provided the research foundation for the SalB to protect the BBB .

Melasma is a common cutaneous disorder of pigment metabolism with complex etiology and pathogenesis.Combination therapy has been preferred by dermatologists for better therapeutic effects and less adverse reactions compared with monotherapy.At present,the treatment of melasma is diversiform,mainly including oral or topical drugs,lasers or photon therapy and combination therapy,etc.Individualized treatment is recommended based on etiology,clinical course and types of melasma,as well as previous treatment history.Combination therapy,sequential therapy or supplement therapy should be included in the treatment of melasma.

Objective To establish a stable in vitro model of blood-brain/tumor barrier (BBB/BTB) simulating in vivo state u-sing the human brain endothelial cells(hBMEC) ,brain pericytes (PC) and U251 glioma cells(U251) .Methods An in vitro model of BBB/BTB was constructed using Transwell inserts(pore size 1 .0 μm)coculture while three cell subculture reaches a certain num-ber .Its barrier function was evaluated by the 4-hour leakage test ,inverted microscope to observe morphological changes ,horseradish peroxidase permeability test and tight junction protein Claudin-5 and Occludin identification expression observed by immunofluores-cence technology .Results hBM EC monolayers demonstrated a typical cobblestone-like appearance ,Non-contact spindle cocultured monolayer growth appeared its uniqueswirl poolshape ,fusiform morphology ;the pericytes displayed irregular shape and overlap-ping grow ;U251 showed typical tumor cell growth .Fluid leakage test were interviewing a certain level difference in three different ways to build models ;immnocytochemical staining showed a continuous and dense tight junction formed between the endothelial cells but intensity ranging ;horseradish peroxidase permeability of three different models showed that pure hBMEC (43 .490 ± 3 .572)% ,hBMEC+U251(36 .540 ± 1 .475)% ,hBMEC+ PC+U251(26 .460 ± 2 .372)% was significantly lower ,the difference was statistically significant(t=19 .330 ,P<0 .01) .Conclusion hBMEC+ PC+ U251 cocultured the morphology ,tructure and barrier function have more advantages than pure hBMEC and hBMEC+U251 ,have the basic characteristics and function of BTB/BBB ,and they have provided a new tool reseach function ,regulation mechanisms and drug screening the BTB/BBB in future .

Objective: To investagate the antioxidative action of aloe and its dose-effect relationship. Methods:Sprague-Dawley rats were killed and then the livers were removed to isolate the microsome which can generate the reactive oxygen species in the presence of VC and Fe2+ or cumene hydroperoxite(CHP). In these peroxidative damage models, different dosages of aloe extract were added. Then the contents of malondialdehyde (MDA) were examined for analyzing the antioxidative action of aloe extract. Results:In CHP model, the content of MDA in those groups with different dosages of aloe extract decreased significantly (P

Objective To evaluate the degree of hypothermia cerebral resuscitation patients by acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and judge the prognosis,so as to prove the effectiveness of its application.Methods Data of 34 cases of patients with mild hypothermia cerebral resuscitation in ECIU or ICU were continuously observed,calculated the scores of APACHE Ⅱ and verify for establishing the regression model.Results The APACHE Ⅱ score was 20-47 (33.86 ± 5.12) scores.The APACHE Ⅱ score in 9 patients who were survived within 72 h was (27.83 ± 4.89) scores,and in 25 patients who were dead within 72 h was (35.56 ± 7.12) scores,there was significant difference (P ＜ 0.01).Logistic regression analysis showed that APACHE Ⅱ score was the risk factor (P ＜ 0.01).Conclusion APACHE Ⅱ score can be applied to evaluate the degree of mild hypothermia cerebral resuscitation and prognosis,it can guide the clinical decision making.

Objective:To predict and analyze the secondary structure and B cell epitopes of Izumo protein.Methods: The secondary structure and flexible regions of Izumo protein were predicted by the methods of Chou-Fasman,Gamier-Robson and Karplus-Schulz.Moreover,hydrophilicity plot,surface probability plot and antigenic index of Izumo protein were predicted by the methods of Kyte-Doolitde,Emini and Jameson-Wolf,respectively.Results: Izumo protein contained moreαhelix regions.There were several centers ofαhelix in the regions of 6-17,30-40,88-99,103-120,153-160,173-188,249-260,283-297,334-338 and 339-346 of Izumo protein,and several centers of βsheet in the regions of 21-25,198-200,245-248 and 320-323.Moreover,many distinct B cell epitopes in Izumo protein possibly localized in the regions of 3642,62-66,94-99,118-122,129-132,151-154,161-164,173-177,205-208,212-216,256-265,271-276,283-288,314-318 and 336-350.Conclusion:These results are helpful for identification of the dominant B cell epitopes and the functional domains of Izumo protein.

The preparation of electrophoretic microcolumn and its application to the electrophoretic separation of amino acids with a 2-mm I.D. fused-silica microcolumn packed with uniform quartz microncrystal prepared by hydrothermal synthesis are reported. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 30% (V/V) methanol, the tryptophan, phenylalanine and tyrosine were separated by the microcolumn electrophoresis and detected by an UV spectrophotometer without derivatization. The limits of detection were 0.038, 0.21 and 0.20 mol/L, respectively. The separation efficiency of tryptophan was 4.4×104 plates/m. The sample capacity of the electrophoretic microcolumn achieved 35 μL. The precisions of the microcolumn electrophoresis were satisfactory. The thermal effects of the electrophoretic microcolumn that without packing, packed with 360 μm quartz sand and with 9 μm length quartz microncrystal were discussed, respectively. It was found that the electrophoretic microcolumn packed with quartz microncrystal was able to inhibit Joule heat, increase sample capacity and enhance detection sensitivity. The microcolumn electrophoresis is one of the high-performance separation techniques for an in-situ, real-time and portable electrokinetic flow analysis system.

Background and objective Targeting the mutations and amplifications in the epidermal growth factor receptor (EGFR) gene has curative effects on cancers of the lung, oral cavity, and gastrointestinal system. However, a systemic immune inflammation is an adverse effect of this therapeutic strategy. In this study, we aimed to identify the possible changes in the tumor microenvironment that contribute to the anti-cancer activity of EGFR inhibition.Methods Squamous-cell cancers were induced by the syngeneic transplantation of either EGFR-null or wild-type mouse primary keratinocytes that had been transduced with an oncogenic H-ras retrovirus. The mice were treated with gefinitib. Then, flow cytometric was used to detect the ratio of T cells and the expression of programmed cell death receptor 1 (PD-1). RT-PCR was used to detect the expression of cytokines and chemokines.Results Tumors that formed from EGFR-null keratinocytes were smaller, had fewer infilltrat-ing FoxP3+ Treg cells, lower Foxp3 RNA, and lower percentage of PD-1 positive CD4 cells than those formed from wild-type keratinocytes. These results indicated that tumor cells can autonomously regulate the tumor microenvironment. Hosts with wild-type cancers and that were treated with gefitinib for 1 week tended to have smaller tumors. The treated mice in the short-term pharmacological model tended to have reduced FoxP3+ cells and FoxP3 RNA in the tumor microenvironment, as well as a substantially increased ratio of IL-1A/IL-1RA transcripts. These results suggested that the brief systemic inhibition of EGFR signaling alters the immune environment of the targeted cancer.Conclusion The autonomous (genetic) or systemic (pharmacologic) inhibition of EGFR signaling in tumor cells reduces tumor growth and Treg infilltration in the tumor micro-environment. An EGFR-dependent Treg function supports the growth of squamous cancers. Therefore, Treg is a target in the therapeutic strategy of EGFR inhibition.

Background and objective Tumor-associated fibroblasts (TAF) is an important part of TME, which inhibits the function of immune cells. CD8+ T cells play a significant role in tumor immunity. T-cell membrane possesses a distinct type of molecule with a negative regulatory function. Upon interaction with its corresponding ligand [programmed death factor ligand 1 (PD-L1)], programmed death factor 1 (PD-1) is activated and thus inhibits the kinase activity of T cells. This study aims to explore the possible effects of TAF on PD-L1 expression in lung cancer cells. Methods Lung cancer cell lines H1975 and H520 were co-cultured with (experiment) or without TAF (control) via Transwell assay for through 48 hours under the same culture condition. H1975 and H520 cells were counted using a microscope. The protein and mRNA expression levels of PD-L1 were detected by FCM assay and PCR analysis, respectively. Results The numbers of lung cancer cells in 100μm2 for H1975 and H520 cells are (46±21) and (38±10) in the experiment group, respectively, and (16±5) and (12±5) in the control group, respectively (P<0.05). The expression levels of the PD-L1 protein in H1975 and H520 cells are (20.93%±3.54%) and (19.26%±3.04%) in the experiment group, respectively, and (12.58%±2.52%) and (11.60%±2.65%) in the control group, respectively (P<0.05). The mRNA expression levels in H1975 and H520 cells are (16.45±1.25) and (15.38±2.02) pg/mL in the experiment group, respectively, and (7.78±1.27) and (7.20±1.58) pg/mL (P<0.05) in the control group, respectively (P<0.05). Conclusion TAF promotes the growth and increases the expression of PD-L1 in H1975 and H520 cells.

Objective To assess the development,progression and change of nonmotor symptoms in patients with Parkinson' s disease and its impact on patients' quality of life.Methods Eighty-seven consecutive patients with idiopathic Parkinson' s disease were studied.Parkinsonian status was assessed at baseline and 3 years follow-up using Unified Parkinson' s Disease Rating Scale (UPDRS) part Ⅲ & Ⅳ,Nonmotor Symptoms Questionnaire (NMSQuset),Parkinson-related quality of life (PDQ) scales.Paired ttest,Chi-square test,Spearman rank order correlation and hierarchical regression of the major statistical procedures were employed.Results At 3 years follow-up,compared to baseline,the UPDRS Ⅲ score (22.21 ±11.31 vs 30.49± 11.68),UPDRS Ⅳ score(1.00±1.54 vs 2.94±3.12),NMS score (7.98±3.96 vs 12.35 ± 5.12) and PDQ score (28.11 ± 22.88 vs 36.65 ± 26.95) were significantly higher ( t =- 5.54,- 5.75,- 6.46,- 5.29,all P =0.000,respectively).The aggravation of motor and nonmotor symptoms caused the decline of quality of life.The prevalence of constipation,problem of remembenng thing,nocturia ranked tops,and depression,and anxiety were still in the middle,compared with baseline.The prevalence of pains,sweating,dribbling,sense of incomplete emptying etc were significantly increased during the follow-up,△R2 were 21.6％ and 23.4％ respectively,resulting in the deterioration of quality of life.Conclusions PD nonmotor symptoms appear from the early stage.The motor and nonmotor symptoms aggravate over time.