Background Immediately ocular rinsing is a key step for the prevention of eye tissue damage after acute chemical bums.A good ophthalmic rinsing solution can neutralize chemical substance and improve the prognosis of patients.Objective This study attempted to evaluate the therapeutic effect of self-made citric aciddisodium hydrogen phosphate buffer and citric acid-disodium hydrogen phosphate-potassium chloride buffer on corneal chemical burns in mice.Methods Citric acid-disodium hydrogen phosphate buffer solution (solution 1) and citric acid-disodium hydrogen phosphate-potassium chloride buffer solution (solution2) with the pH 7.4 were prepared.One hundred and twenty clean male C57 mice aged 6-8 weeks were randomized to two groups,and filter papers containing 1 mol/L H2SO4 or 0.15 mol/L NaOH were attached to the central corneas of the right eyes to create the acid or alkali burning models.Then the eyes were immediately rinsed by 40 ml solution 1,tap water or solution 2 according to the grouping and the model eyes without rinsing served as the control group.The corneal opacity was examined by slit lamp microscope and scored in 3,7 and 14 days after modeling.The percentages of corneal fluorescein staining,corneal neovascularization and corneal ulcer were analyzed.The study protocol was approved by Experimental Animal Ethic Commission of Second Military Medical University.Results In the corneal acid burning models,the number of eye with corneal opacity scored 1 in the solution 1 group,tap water group and solution 2 group was significantly more than that in the non-rinsing group in 3,7 and 14 days after modeling (all at P＜0.01);In 3 days after modeling,the numbers of eye scored 1 were more in the solution 1 group than those in the tap water group and solution 2 group (x2 =11.000,P =0.001;x2 =4.000,P =0.046).There were no differences in the eye number of different corneal opacity scores in 14 days after acid burning (all at P＞0.05).In 3,7 and 14 days after corneal alkali burning,the number of eyes with corneal opacity scored 1-2 was significantly increased in the solution 1 group,tap water group and solution 2 group compared with non-rinsing group (all at P＜0.01).The percentage of corneal ulcer in the solution 1 group,tap water group and solution 2 group was 7％,27％ and 13％,respectively,which was significantly lower than 73％ in the non-rinsing group (P =0.000,0.027,0.003),and no significant differences were seen in various time points after corneal alkali burning (all at P＞O.05).Corneal neovascularization occurred in 50％ mice in non-rinsing group in 14 days after acid burning.However,no neovascularization was seen in the mice of the solution 1 group,tap water group and solution 2 group in both acid and alkali burning mice.Conclusions Citric aciddisodium hydrogen phosphate buffer (pH 7.4) appears to be an effective emergency rinsing solution for corneal chemical burns and the rinsing solution with or without potassium chloride is not obviously affected to the prognosis of corneal chemical burns in the mice.

Objective To observe the level of dopamine and its receptor in striatum from conditioned place preference (CPP) of rats administrated of morphine.And explore the mechanism of opioid-psychic dependence involving the levels of neurotransmitter and receptor.Methods CPP model was validated in morphine-dependence rats for 10 days.Striatum samples were harvested from two separate groups involving the saline group and morphine treated group (n =10 per experiment).The DA contents in striatum were detected in rats with colorimetry.Immunohistochemistry was applied to detect the differential expression levels of dopamine receptor 2 (DRD2)in samples.Results Compared with the saline group,higher content of neurotransmitter dopamine was observed in the morphine dependent rats (7.63 ±0.98 vs 5.23 ± 1.01,P＜0.01),while the expression level of DRD2 was down regulated (0.06 ± 0.02 vs 0.08 ± 0.02,P ＜ 0.01).Conclusion The increased expression of neurotransmitter dopamine and the decreased of DRD2 may be contributed to morphine-induced psychological dependence in morphine dependent rats.

Objective:To observe the killing effect of irradiated peripheral blood mononuclear cells (PBMCs) at low dose combined with apogossypolone (ApoG2) on cultured human prostate cancer PC-3 cells.Methods:Human PBMCs were irradated by gamma ray at 1 gray,the irradiation dose rate was 17 Gy/min.The experiment were divided into PC-3 tumor cell control group,PC-3 cells with irradiated and non-irradiated PBMCs co-culture groups,ApoG2 treatment group,irradiated PBMCs and ApoG2 co-treatment group.Acridine orange/ethidium bromide (AO/EB) staining and MTT method were used to observe the killing effect of PBMCs and/or ApoG2.Results:The killing activity of irradiated PBMCs group and ApoG2 treatment group were obviously increased and were higlaer than that of non-irradiated group (P＜0.05).The killing activity of combined group were much higher than that of irradiated group and ApoG2 treatment group (P ＜0.01 ).Conclusion:Irradiated PBMCs at low dose combined with ApoG2 can enhances the anti-tumor effects markedly.

Objective To observe effects of conditioned place preference (CPP) of morphine dependent rats,variation of dopamine neurotransmitter and its receptor 2 of the striatum in rats suffered from 1-tetrabydropalmatine(l-THP).Methods The CPP model was established by morphine injection in rats with a increasing dose for 10 days,with the initial dose of 10 mg · kg-1 and the final dose of 100 mg · kg-1,10 mg · kg-1 was increased each day,thus 100 nmg · kg-1 Was injected by day 10.Treatments with administration of 1-THP(3.76,1.88 and 0.94 mg/kg) were performed respectively for 6 days,and the effects of CPP for psychological dependence in these rats were observed.Striatum samples were taken out and their variable contents of dopamine neurotransmitter and its receptor 2 in striatum were detected by high performance liquid chromatography and immunohistochemisty,respectively.Results Compared with the NS treatment group,the time of animals treated with 1-THP (3.76 and 1.88 mg/kg) staying in drug-paired compartment were reduced (354 ± 58,373 ± 79) (P＜ 0.01) ;the raised of variation contents of dopamine neurotransmitter were reduced in striatum from those rats(5.49 ± 1.95,6.11 ± 1.05),while the expression level of dopamine receptor 2 was increased(0.08 ± 0.02,0.07 ± 0.03) (P ＜ 0.01 or P ＜0.05).Conclusion Reversing dopamine neurotransmitter and its receptor 2 in striatum of morphine dependent rats may be one of the possible mechanisms that 1-THP effectively inhibit the effects of morphine CPP.

Objective To investigate expression level of endoplasmic reticulum stressmarker GRP78 in the testicular tissue in rats with different phases of morphine-dependence. To explore the role of ERS in morphine-de-pendence. Methods SD rats were divided into 6 groups: morphine (mor) -withdrawal group, mor-extinct group, mor-kindling group and their control groups, normal saline (NS)-withdrawal group, NS-extinct group, NS-kindling group. The experimental rats were injected with morphine subcutaneously on increasing dosage to establish the con-ditioned place preference (CPP) model. The rats in control groups were injected NS. Then the rats were suffered from withdrawal for 48 h, extinction and kindling by morphine, separately. The GRP78 expression level in testicular tissues of rats in the time point mentioned above were measured using Western Blot. Results The time of rats in the paired-box was (528.0 ± 81.0) s, which was significantly higher than that in the NS control group (P<0.001). It was (396.8 ± 116.9) s after extinctive phase, which was significantly higher than that the withdrawal phase of rats (P < 0.001). Also it was (396.8 ± 116.9) s after kindling with morphine which was significantly higher than that the extinctive phase of rats (P < 0.001). These changes of the time indicated that the animal models of extinction and kindling were established in the study. The GRP78 levels were down-regulated in 48 h after withdrawal (P <0.05), and increased a bit afterextinctive phase, but up-regulated highly after kindling with morphine (P < 0.01). Conclusion ERS may be related in the morphine dependence and it might play an important role of testicular dys-function in male drugabuser.

Objective To find the different plasma-associated proteins of rheumatoid arthritis (RA) by using two-dimensional gel electrophoresis for understanding the pathogenesis of RA. Methods The total protein from either RA patients or normal ones was prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis. After silver staining, gel-image analysis was performed by using PDQuest. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Results 2-DE patterns of plasma from controls and RA patients were presented. The results showed that average number of protein spots was 592 and 563 respectively, and the corresponding average matching rate was 89% and 87% respectively. Gel-image analysis revealed that there were 24 differential protein spots. A total of 15 differential protein spots were successfully identified by MALDI-TOF-MS, of which 6 proteins were up-regulated as compared with control. Conclusion The differentially expressed proteins can be observed in plasma from RA and controls, which can be used to elucidate the pathogenesis of RA for further study.

Objective To screen the proteins interacting with FOXP3 in yeast two-hybrid system. Methods The "bait plasmid" pGBKT7 (named as pGBKT7-FOXP3) was constructed successfully. Using FOXP3 as bait, a human liver cDNA library was screened and the proteins interacting with FOXP3 were searched. The false positive clones were discarded by one to one yeast two-hybrid system, and the positive clones were sequenced and analyzed by bioinformatic methods. Results The bait plasmid pGBKT7-FOXP3 was constructed successfully and there was no self-activation or toxicity in AH109. Three proteins had been found in our system to be able to interact with FOXP3. They were tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein. Conclusion FOXP3 interacts with tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein, all of which may interfere in cell metabolism and function of T cell.

Objective To establish an infrared spectrophotometric method for determination of mineral oil mist in workplace air. Methods The mineral oil mist in workplace air was sampled with glass fiber filter membrane and eluted with carbon tetrachloride. Petroleum-like standard solution of carbon tetrachloride was used as the calibration standard, and quantitative analysis was performed using infrared spectrophotometric oil analyzer. Results The sampling efficiency of the glass fiber filter membrane ranged from 94.8% to 99.2%, and the extraction efficiency ranged from 95.6% to 104.2%. The linear range of mineral oil mist was 1.00-120.00 mg/L, with a correlation coefficient of 0.999 4. The detection limit was 0.52 mg/L, and the quantification limit was 1.74 mg/L. The average recovery rate ranged from 98.8% to 104.1%. The within- and between- run relative standard deviations were 2.2%-6.4% and 2.3%-5.2%, respectively. The samples were stable at room temperature for seven days. This method could be used for air sampling of mineral oil mist in workplaces where mineral oil is used. Conclusion The method is sensitive, accurate, and efficient, which is suitable for determining the concentration of mineral oil mist in workplace air.

Objective To construct a bait vector containing human Foxp3 gene in yeast two-hybrid system in order to screen the cDNA library of T lymphocyte. Methods RT-PCR was used to amplify the Foxp3 gene fragment from the peripheral blood mononuclear cells (PBMC) with the primers designed in accordance with the sequence in GenBank. The product was inserted into pMD18-T vector. After verified with restriction endonuclease digestion of EcoRⅠ and SalⅠ, the vector was inserted into the “bait plasmid” pGBKT7 (named as pGBKT7-Foxp3). After confirmation with restricted endonuclease digestion and sequence analysis, the plasmid was transformed into the yeast cell AH109, and its toxicity and transcriptional activation was tested by both the phenotype assay and the color assay. Results The amplified product of 1 203 bp was inserted into PMD18-T vector and proven correctly by double restriction enzyme digestion. Sequence analysis revealed that the fragment was correctly inserted into pGBKT7 with a right reading frame and its expression in yeast was verified. Conclusion The bait plasmid pGBKT7-Foxp3 constructed expresses correctly, and can not activate the transcription of reporter gene alone in yeast two-hybrid system

ObjectiveTo predict the potential targets and possible related signaling pathways of Salviae Miltiorrhizae Radix et Rhizoma against bladder cancer （BC） based on network pharmacology and verify the potential molecular mechanism through in vitro cell experiment. MethodActive components of Salviae Miltiorrhizae Radix et Rhizoma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and BC-related targets were searched from GeneCards and Online Mendelian Inheritance in Man （OMIM）. Via Venny2.1， the potential targets of Salviae Miltiorrhizae Radix et Rhizoma against BC were screened out and the Venn diagram was plotted. Protein-protein interaction （PPI） network was constructed by STRING， followed by Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Gnomes （KEGG） pathway enrichment with DAVID. Cell Counting Kit-8 （CCK-8） assay was employed to detect the inhibitory effect of tanshinone Ⅱ_A （Tan Ⅱ_A）， cryptotanshinone （CPT）， and luteolin （LUT） at different concentration （0， 1， 2， 4， 8， 16， 32 μmol·L^-1） on the proliferation of BC T24 and 5637 cells， propidium iodide （PI） staining to analyze the apoptosis of 5637 cells induced by Tan Ⅱ_A， CPT， and LUT （0， 4， 8 μmol·L^-1）， and Western blotting to detect the regulatory effect of Tan Ⅱ_A （0， 4， 8， 16 μmol·L^-1） on the expression of key target proteins. ResultA total of 65 active components and 39 anti-BC targets of Salviae Miltiorrhizae Radix et Rhizoma were screened out. The anti-BC targets were mainly involved in the KEGG pathways of neuron-ligand-receptor interaction， phosphatidylinositol 3-kinases （PI3K）/protein kinase B （Akt） signaling pathway， epidermal growth factor receptor （EGFR） tyrosine kinase inhibitor resistance， and hypoxia inducible factor （HIF）-1 signaling pathway. As for the CCK-8 assay， compared with the blank group， Tan Ⅱ_A， CPT， and LUT significantly inhibited the proliferation of T24 and 5637 cells， particularly the 5637 cells. The half maximal inhibitory concentration （IC₅₀） of Tan Ⅱ_A on 5637 cells was significantly lower than that of CPT and LUT. Moreover， compared with the blank group， Tan Ⅱ_A， CPT， and LUT all induced the apoptosis of 5637 cells， and the effect followed the order of Tan Ⅱ_A>CPT>LUT （P<0.05）. Western blot showed that Tan Ⅱ_A significantly reduced the expression of EGFR， p-PI3K， and p-Akt in 5637 cells in a concentration-dependent manner compared with the blank group （P<0.05）. ConclusionSalviae Miltiorrhizae Radix et Rhizoma exerts therapeutic effect on BC through multiple components， multiple targets， and multiple pathways. The mechanism is the likelihood that it down-regulates the expression of EGFR， p-PI3K， and p-Akt proteins， thus further inhibits cell proliferation， and induces apoptosis.