Objective:To explore the homing effect of mouse bone marrow-derived mesenchymal stem cells (mBMSCs) on the tongue squamous cell carcinoma Cal27 cells, and to observe the biological behaviors of tongue squamous cell carcinoma Cal27 cells affected by mBMSCs. Methods:The mBMSCs were isolated and purified by whole bone marrow methods. The cell phenotype was analyzed by flow cytometry to identify the mBMSCs. The mBMSCs were divided into blank control group, Hacat cell control group and Cal27 cell experiment Cal27 cell group. After co-cultured with normal culture medium, Hacat conditioned medium and Cal27 conditioned medium,the number of the migrated Cal27 cells was detected by cell migration assay. The Cal27 cells were divided into single Cal27 cell control group and co-cultured Cal27 and mBMSCs experiment group(co-cultured group). The Cal27 cells in co-cultured group were co-cultured with mBMSCs using Transwell method. Cell proliferation assay and colony formation assay were used to detect the proliferation ability of the co-cultured Cal27cells. Cell migration assay was performed for the detection of the migration ability of the Cal27 cells after co-culture. Results:The inverted microscope results showed that most mBMSCs were short shuttle-like, with abundant cytoplasm, oval or nephron nucleus and irregular cell lengths; the cells arranged well. The results of flow cytometry showed that CD44, CD29, and Sca-1 were positive, and CD45, CD11b were negative in the mBMSCs. Compared with blank control group and Hacat cell control group, the number of mBMSCs homing to Cal27 cells in Cal27 cell group was significantly increased (P<0.01). Compared with single Cal27 cell control group, the growth rates of Cal27 cells in co-cultured group were significantly increased from the 3rd day after co-culture(P<0.01), and the number of clones was increased. The cell migration test results showed that the number of transmembrane cells in co-cultured group was significantly increased compared with single Cal27 cell control group (P<0.01). Conclusion:mBMSCs can home to the tongue squamous cell carcinoma Cal27 cells, which promotes the proliferation and migration of tongue squamous cell carcinoma Cal27 cells and promotes the development of tongue squamous cell carcinoma. So mBMSCs can be used as an anti-tumor target.

Objective:To investigate the inhibitory effect of resveratrol on the pulmonary inflammation in the mice with neutrophilic asthma, and to elucidate its mechanism. Methods:A total of 18 female C57BL/6J mice were selected.The mouse models of neutrophilic asthma were established by sensitization with lipopolysaccharide (LPS) and ovalbumin (OVA). The model mice were randomly divided into model group (LPS + OVA group), resveratrol group(Res group, intragastric administration of 30 mg·kg^-1 resveratrol at 2 h before challenge) and N-acetylcysteine group (NAC group, intragastric administration of 3 mmol·kg^-1 N-acetylcysteine at 2 h before challenge). Other 6 age-matched female C57BL/6J mice were choosen as normal control group (PBS group). The behavioral changes of mice in various groups were observed during the atomization with methacholine; the airway hyperresponsiveness (AHR) of mice was analyzed with a lung function instrument, and the total number of cells and inflammatory cells in the bronchoalveolar lavage fluid (BALF) were observed with light microscope; the pathological changes of lung tissue were observe by HE staining; the levels of interleukin-17(IL-17) in the supernatant of BALF of the mice in various groups were detected by enzyme-linked immunosorbent assay (ELISA); the level of reactive oxygen species (ROS) in lung tissue was analyzed by specific fluorescent probe DCF-DA staining; the level of malondialdehyde (MDA) in lung homogenate was measured by MDA kit. Results:Compared with PBS group, the Penh value, total number of cells, inflammatory cells, airway inflammation and scores of the mice in LPS+OVA group were significantly increased (P<0.05 or P<0.01); the IL-17 level in BALF supernatant was significantly increased (P<0.01); the ROS fluorescence intensity in lung tissue was significantly increased, and the MDA level in lung homogenates was significantly increased (P<0.01). Compared with LPS + OVA group, the Penh value, total number of cells, inflammatory cells, airway inflammation and scores in Res group and NAC group were significantly decreased (P<0.05 or P<0.01); the IL-17 levels in BALF supernatant were significantly decreased (P<0.05 or P<0.01); the ROS fluorescence intensities in lung tissue were decreased, and the MDA levels in lung homogenates were significantly decreased (P<0.01). Conclusion:Resveratrol can effectively inhibit the inflammatory response of lung tissue in the mice with neutrophilic asthma, and its mechanism may be related to inhibiting the oxidative stress reaction.

Objective:To observe the apoptosis and mitochondrial fission of human ovarian cancer SKOV3 cells after treated by myricetin and dynamin related protein 1 (DRP1) inhibitor mdivi-1 alone or combined, and to explore the mechanism of myricetin in inducing the apoptosis of SKOV3 cells. Methods:The SKOV3 cells were cultured in vitro and randomly divided into control group, mdivi-1 group, myricetin group and combined group.The cells in mdivi-1 group were treated with 50 μmol·L^-1 madivi-1 for 1 h followed by common culture medium for 23 h;the cells in myricetin group were treated with 50 g·L^-1 myricetin for 24 h;the cells combined group were treated with 50 μmol·L^-1 midiv-1 for 1 h followed by 50 g·L^-1 myricetin for 23 h. The survival rates of cells in various groups were detected by MTT assay. The apoptoic rates of cells in various groups were detected by Muse^® apoptosis detection kit. The expression levels of Cyt C, caspase3, DRP1 and FIS1 were observed by Western blotting method. The mitochondrial fission of cells in various groups was observed with MitoTracker^® Red. Results:Compared with control group, the survival rate of cells in myricetin group was decreased significantly (P<0.05); compared with myricetin group, the survival rate of cells in combined group was increased significantly(P<0.05). Compared with control group, the apoptotic rate of cells in myricetin group was increased (P<0.05); compared with myricetin group, the apoptotic rate of cells in combined group was decreased (P<0.05). Compared with control group, the expression levels of Cyt C and caspase3 proteins in the cells in myricetin group were increased(P<0.05); compared with myricetin group, the expression levels of Cyto C and caspase3 proteins in the cells in combined group were decreased(P<0.05). Compared with control group,the degree of mitochondrial fission of the cells in myricetin group was increased;compared myricetin group,the degree of mitochondrial fission of the cells in combined group was decreased.Compared with control group, the expression levels of DRP1 and FIS1 proteins in the cells in myricetin group were increased (P<0.05); compared with myricetin group, the expression levels of DRP1 and FIS1 proteins in the cells in combined group were decreased (P<0.05). Conclusion:Myricetin can induce the apoptosis of human ovarian cancer SKOV3 cells by promoting the DRP1-dependent mitochondrial fission.

Objective:To explore the influence of interleukin-33(IL-33) on the abilities of spatial learning and memory and the levels of amyloid β-protein(Aβ) and helper T cell 2 (Th2) in the model rats with Alzheimer's disease (AD),and to clarify the effect of IL-33 in the scavenging of Aβ in brain tissue of the model rats with AD and its mechanism. Methods:A total of 70 SD male rats were randomly divided into normal control group (intragastrical administration of double steaming water and subcutaneous injection of D-galactose),model group,sham operation group and extremely low dose,low dose,middle dose,high dose(0.01,0.10,1.00,and 10.00 mg·L^-1) of IL-33 groups (intragastrical administration of AlCl₃ and subcutaneous injection of D-galactose);10 rats in each group. The agents were administered once daily for 60 d. After the last administration,step down test and Morris water maze experiment were used to detect the latency,the mistake times,the escape latencies,the resident time in target quadrant,and the swimming distance. After the behavioral testing,IL-33 was injected intracerebroventricularly for each dose of IL-33 group,and saline was injected intracerebroventricularly in the rats in sham operation group. ELISA method was used to detect the levels of Aβ and Th2 in brain tissue of the rats. Results:The results of step down test showed that the latency of the rats in model group was obviously shortened (t=8.154,P<0.01) and the mistake times were significantly increased compared with normal control group(t=4.579,P<0.01).The results of Morris water maze test showed that the escape latency was obviously prolonged(t=27.810,P<0.01),the resident time in target quadrant and the distance of swimming of the rats in model group were significantly reduced compared with normal control group(t=3.767,P<0.01;t=1. 973,P<0.05).The ELISA results showed that the level of Aβ in brain tissue of the rats in model group was significantly increased (t=3.222,P<0.05) compared with normal control group,and the levels of Aβ in the brain tissue of the rats in low,middle and high doses of IL-33 groups were significantly reduced compared with model group (P<0.05).The level of Th2 in brain tissue of the rats in model group was significantly decreased compared with normal control group(t=4.646,P<0.01), and the levels of Th2 in brain tissue of the rats in low,middle and high doses of IL-33 groups were significantly increased compared with model group (P<0.05). Conclusion:IL-33 has a scavenging effect on Aβ in brain tissue of the AD model rats,and its mechanism may be related to the increase of Th2level in brain tissue of the rats.

Objective:To explore the influence of plateau environment in the development and morphology of the main organs of the offsprings of migrated rats, and to observe the pathological changes of heart, brain, and lung tissues of the offsprings of migrated rats. Methods:The 8-week Wistar rats who lived in the plain area were migrated to the plateau area. After 1 week, a total of 56 female and male rats were fertilized with a ratio of 3:1, and all the pregnant rats were natural childbirth. The offsprings rat pups were divided into three groups:1 month, 3 months, and 6 months. Ten offsprings (5 female and 5 male) were randomly selected in each group to collect the heart, brain, lung, liver, kidney and other major organs to measure the weights. In each group, 45 offsprings were randomly selected to conduct water maze test, open field test and test of captive reaction. The brain, heart, and lung tissues from 5 offsprings in each group were collected for tissue section and the pathological changes of above organ tissues were detected with HE staining. Results:The pregnant rats moved from the plain to the plateau had normal feeding behavior, without preterm birth or death. On average, the pregnant rats had 8 to 10 babies per litter, with a total of 345 offsprings. The weight gain of offspring was about 1.0-1.5 g per day. Some of the offsprings had low intake and difficulty in foraging, and 15 offsprings died 3-5 d after birth, with a mortality rate of 4.3%.The weight and the ratio of liver weight to body weight of the 6-month-old offsprings were increased compared with the 1-month-old offsprings (P<0.05). In Morris water maze test there were no drowning and death in 45 offsprings at different time points; the time to find the water platform of 7 offsprings in 3-month-old group was lower than the others (P<0.05),but there were no significant differences between various groups at other time points(P>0.05).In open field test,there were 6 offsprings in open fieled test in 1-month old group showed a longer stay time in the central region than other rats(P<0.05);there were no significant differences between various groups at the other time points(P>0.05). In spite of all the time points in the captive reaction experiment, all the animals behaved in a similar way. The HE staining results of myocardium tissue showed the myocardial inflammatory cell infiltration,venous congestion, nucleus disappearance,and visible cellular outline in the 1-3 month old offsprings and the myocardial vascular congestion and myocardial space enlargement in the 6-month-old offsprings. The HE staining results of brain tissue showed the glass body formation and nucleus disappearance in the 1-month-old offsprings and neuronal cell body deformation, blood vessel congestion and vacuolar degeneration in the 3-6-month-old offsprings. The HE staining results of lung tissue showed the thicker alveolar walls, lymphocyte and plasma cell infiltration, pulmonary capillary expansion and congestion in the 1-3-month-old offsprings and the thicker alveolar walls,lymphocyte and plasma cell infiltration,pulmonary capillary expansion and congestion, pulmonary interstitial edema and red blood cell liquefaction in the blood vessels in the 6-month-old offsprings. Conclusion:Tibetan plateau environment has an influence in the development and morphology of heart, brain and lung of the migrated rats, and the reason may be related to low pressure and hypoxia of plateau.

Objective:To detect the expressions of inositol-requiring enzyme-1α(IRE1α) and X box-binding protein-1(XBP1) mRNA and proteins in mouse testis cells,and to explore the role of IRE1-XBP1 pathway activation in the endoplasmic reticulum stress induced by low dose radiation(LDR) in the mouse testis cells. Methods:A total of 50 heatly male ICR mice were randomly divided into 10 groups.After the mice were irradiated with 75 mGy X-ray in whole body at different time (0,3,6,12,and 24 h) or with different doses(0,50,75,100,and 200 mGy) of X-ray for12 h,the expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA and proteins were measured by real-time quantitative PCR and Western blotting method,respectively. Results:The expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA (except T-XBP1 and S-XBP1 mRNA at 3 h after irradiation) in the testis cells of the mice after irradiated with 75 mGy X-ray for 0-24 h were increased with the time prolongation,and reached to the maximum value at 6 h after irradiation,then were gradually decreased.Compared with 0 h group,the expression levels of IRE1α mRNA(6,12, and 24 h after irradiation),T-XBP1 mRNA and S-XBP1 mRNA(6 and 12 h after irradiation) were significantly increased (P<0.05 or P<0.01); the expression intensities of IRE1α and S-XBP1 proteins were increased; the expression intensities of IRE1α (6 and 12 h after irradiation)and S-XBP1(24 h after irradiation) proteins were increased most significantly,but the expression intensity of T-XBP1 protein was slightly decreased. The expression levels of IRE1α,T-XBP1 and S-XBP1 mRNA in the testis cells of the mice after irradiated with 0-200 mGy for 12 h were increased,and reached to the maximum values after 75 mGy irradiation,then they were gradually decreased,even below 0 mGy. Compared with 0 mGy group,the expression levels of IRE1α mRNA in the testis cells of the mice in 75 and 200 mGy groups and the expression levels of T-XBP1 mRNA and S-XBP1 mRNA in 75 mGy group were significantly increased (P<0.05 or P<0.01); the expression intensities of IRE1α protein in 75 mGy group and S-XBP1 protein in 75 and 200 mGy groups were increased mostly,while the expression intensity of T-XBP1 protein had no obvious change. Conclusion:LDR can induce the increase of IRE1α and S-XBP1 mRNAs and proteins,and activate the IRE1-XBP1 pathway in endoplasmic reticulum stress.

Objective:To investigate the regulatory effect of genipin (GP) on the gastric cancer SGC 7901 cells, and to clarify its mechanism. Methods:The SGC 7901 cells in logarithmic growth phase were selected and divided into control group and different concentrations (5.0, 10.0, and 20.0 mg·L^-1) of GP groups.MTT was used to detect the inhibitory rates of proliferation of SGC 7901 cells at different time points(24, 48, and 72 h).Transwell chamber cell invasion assay was used to detect the invasion ability of SGC 7901 cells in vitro.The expression levels of Bcl-2, Bax and caspase-3 proteins in the SGC 7901 cells were detected by Western blotting method. Results:The results of MTT assay showed that the inhibitory rates of proliferation of the cells in different concentrations of GP groups after treated with GP for different time were increased significantly compared with control group(P<0.01).The Transwell cell invasion assay results showed that compared with control group,the number of transmembrane cells in 10.0 and 20.0 mg·L^-1 GP groups was decreased significantly after treated for 72 h(P<0.01).The results of Western blotting method showed that compared with control group,the expression level of Bcl-2 protein in 20.0 mg·L^-1 GP group was decreased after treated for 72 h(P<0.01),and the expression levels of caspase-3 and Bax proteins were increased (P<0.05). Conclusion:GP can inhibit the abilities of proliferation and invasion in vitro of SGC 7901 cells, and induce the apoptosis;its mechanism may be related to the regulation of Bcl-2, caspase-3, and Bax protein expressions.

Objective:To screen the genes may be regulated by NOB1 by using gene microarray technique, and to clarify the regulatory effect of NOB1 on the expression of osteosarcoma cell-related genes. Methods:The U2OS cells were treated with lentivirus-mediated RNA interference and to establish the osteosarcoma cells Lv-shNOB1-U2OS. Lv-shCon-U2OS group and Lv-shNOB1-U2OS group were set up. The mRNA expressions of those cells were detected using expression pattern analysis. Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to predict the global functions of NOB1, including biological processes, cellular components, molecular functions, and signaling pathways. Results:After NOB1 interference in the U2OS cells, there were 792 genes with up-regulated mRNA expression level and 1 059 genes with down-regulated mRNA expression level, with a total variation of 1 851 genes. The GO analysis results showed that from the enrichment degree of cell location entries, the differentially encoded product proteins were mainly distributed on the cell membrane as 56.9% of the total difference genes, 39.4% of the totally differential genes distributed in the extracellular region, and 20% of the totally differential genes in the extracellular space; from the enrichment degrees of the molecular function items, the main function of the differentially encoded product proteins was calciu mion binding-related function(22% of the totally differential genes), the second function was transporter activity (9.2% of the totally differenital genes), and the third function was actin binding activity(8.7% of the totally differential genes). In terms of the enrichment of biochemical process entries, the main participation process of differentially encoded product proteins was 18.7% of the totally differential genes of signal transduction, the second involved process was 15.6% of the totally differential genes produced by multiple organelles, and the third process was the cell adhesion process accounted for 10.2% of the totally differential genes. The KEGG analysis results showed that their encoding proteins were involved in plasma membrane, calcium ion binding activity and signal transduction. Conclusion:Knockout of NOB1 can affect the expressions of osteosarcoma cell-related genes in an all-round way.

Objective:To explore the influence of hypoxia and transforming growth factor β (TGF-β) on the differentiation of umbilical cord mesenchymal stem cells (UCMSC) into the type Ⅱ alverolar epithelial cells(ATⅡ) and the regulatory effect of miR-145 in this process, and to illuminate the differentiation mechanism of UCMSC into ATⅡ under the double stimulation of both hypoxia and TGF-β in the damaged lung tissue microenvironment. Methods:The UCMSC were isolated in vitro and co-cultured with human lung cancer cell line A549 to induce the differentiation of UCMSC into ATⅡ. Cobaltous chloride (CoCl₂) was used to mimic the hypoxia condition, and the induced cells were divided into normoxia group and hypoxia group. In hypoxia group, phase contrast microscope was used to observe the changes in cell morphology and flow cytometry was used to detect the percentage of ATⅡ in the induced cells. qPCR and Western blotting methods were applied to test the expression levels of ATⅡ specific genes, fibrosis-related genes and miR-145 in normoxia group and hypoxia group. In hypoxia group, the cells were divided into inh-145 group, inh-145 scramble group and control group, the expression levels of fibrosis-related genes in the cells in three groups were tested by qPCR and Western blotting methods. The pre-145 and pre-145 scramble were transfected into the 293T cells, and then the dual luciferase reporter gene system was used to check the relative luciferase unit (RLU) of TGF-βRⅡ3'-UTR wild type and mutants. Results:In hypoxia group, the fibroblast-like UCMSC became flat and spindle, and finally showed a morphology of cobblestone-like epithelial cells 8 d later, and the percentage of the induced cells with high expression of ATⅡ surface marker SpC was up to (94.50±3.37)%. The expression levels of specific genes of ATⅡ (KGF, CK18, SpA, SpB and SpC) and miR-145 were obviously up-regulated in hypoxia group compared with normoxia group (P<0.05). After stimulating the UCMSC-ATⅡ differentiation with TGF-β1, the expression levels of fibrosis-related genes Col-Ⅰ, TGF-βRⅡ and its downstream signaling factors p-Smad2 and p-Smad3 in hypoxia group were significantly down-regulated compared with normoxia group (P<0.05). The expression levels of Col-Ⅰ and TGF-βRⅡ in inh-145 group were significantly raised compared with inh-145 scramble group and control group under hypoxia induction (P<0.05). The RLU of TGF-βRⅡ 3'-UTR wild type was decreased after treated with pre-145 compared with TGF-βRⅡ mutants (P<0.05). Conclusion:Hypoxia can promote the differentiation of UCMSC into the ATⅡ and inhibit the TGF-β1-induced fibrosis. Its mechanism may be related to the inhibition of TGF-β1/TGFβRⅡ signaling pathway through the down-regulation of TGF-βRⅡ expression by hypoxia-induced miR-145.

Objective:To investigate the regulatory effects of resveratrol (Res) on the epithelial to mesenchymal transition (EMT), migration and invasion abilities of the glioma U87 cells, and to clarify the inhibitory effect of Res on EMT of the cells. Methods:The glioma U87 cells were divided into control group (no treatment), EMT group (EMT was induced with TGF-β1) and Res treatment group(EMT was treated with Res). The expression levels of mesenchymal markers (N-cadherin, β-catenin, Vimentin), EMT-inducing transcription factors (Snail, Slug, Twist1), matrix metalloproteinase (MMP)-2 and MMP-9 in the glioma U87 cells were measured by Western blotting method. Scratch-healing assay and Transwell assay were performed to examine the cell migration and invasion abilities. Results:Compared with control group, the protein expression levels of mesenchymal markers (N-cadherin, β-catenin, Vimentin), EMT-inducing transcription factors (Snail, Slug, Twist1), and MMP-2 and MMP-9 in EMT group were significantly increased (P<0.05).Compared with EMT group, the expression levels of above proteins in Res treatment group were markedly down-regulated(P<0.05 or P<0.01);the effect in 40 μmol·L^-1 Res treatment group was more obvious than that in 20 μmol·L^-1 Res treatment group. The migration and invasion rates of the U87 cells in EMT group were obviously elevated compared with control group(P<0.01), and the migration and invasion rates of the glioma U87 cells in Res treatment group were significantly lower than those in EMT group(P<0.01). Conclusion:Res can inhibit EMT of the glioma U87 cells and reduce the migration and invasion abilities of the cells.

Objective:To discuss the bond strengths of three kinds of metal bottom plate brackets commonly used in clinic and after sandblasting treatment under the condition of artificial saliva,and to evaluate their bonding properties. Methods:A total of 60 premolar teeth extracted because of orthodontic treatment were collected and divided into domestic Xinya bracket group(Xinya group),imported ultra thin MBT bracket group(MBT group),and Japan TOMY lock bracket group(TOMY group)(n=20). The teeth were randomly divided into six groups after the three types of brackets fall off. The three types of new and sandblasting brackets were respectively bonded to the teeth randomly with 10 brackets in each group.The shear strength was detected by universal electronic mechanical testing machine, and the adhesive remnant index (ARI) of enamel surface in each group was obsereved,and the morphological features of three different base brackets and three shedding of brackets conducted by sandblasting were obsereved by scanning electron microscope. Results:Under the condition of artificial saliva,for the first bonding and bonding again,the shear strengths of the new brackets in TOMY goup were higher than those in Xinya group and MBT group (P<0.05),while the shear strengths had no statistical differences between Xinya group and MBT group (P>0.05).For the brackets treated with sandblasting after shedding in three groups, the shear strength in TOMY group was greater than those in Xinya group and MBT group (P<0.05),while the shear strengths in Xinya group and MBT group had no statistical difference(P>0.05). The shear strengths of brackets that were conducted by sandblasting after falling off in TOMY group and MBT group were increased compared with the original brackets(P<0.05).There were no statistical differences in ARI between various groups(P>0.05).The scanning electron microscope results showed that three kinds of brackets were crisscroped with the mesh,and they were more dense in TOMY group. After sandblasting, the sand grains were embedded in the grid,and the sand grains were more embedded in the inverted pits in TOMY group and MBT group. After sandblasting again,the sand particle embedding scope was increased,and the grid structure of the network bottom in Xinya group was destroyed,but there were no obvious abnormities in MBT group and TOMY group. Conclusion:The bond strengths of the three types of bottom plate brackets can meet the clinical needs,and the bond strength in TOMY group is superior to those in Xinya group and MBT group. The use of sandblasting to deal with the shedding of TOMY and MBT brackets can improve the bond strengths of the brackets.

Objective:To observe the effects of myricetin on the proliferation, apoptosis, migration and invasion of mouse glioma GL261 cells, and to provide theoretical basis for myricetin in treating glioma. Methods:The mouse glioma GL261 cells were cultured in vitro. The experiment was divided into control group and different concentrations (10, 20, 30, 40, 50 and 60 μmol·L^-1) of mytricetin groups. CCK-8 assay was used to detect the survival rates of the mouse glioma GL261 cells in various groups.The cell cycle of GL261 cells in various groups was detected by flow cytometry; Hoechst 33342 staining and flow cytometry were used to detect the apoptosis of Gl261 cells in various groups. The number of migrated GL261 cells and the number of GL261 cells entering the lower chamber were detected by Transwell chamber assay. The expression levels of matrix metalloproteinases(MMPs) mRNA were detected by RT-PCR method. Results:The CCK-8 assay results showed that the survival rate of GL261 cells in 20 μmol·L^-1 myricetin group was significantly decreased compared with control group (P<0.01). The flow cytometry results showed that the proportion of GL261 cells in G₁ phase in 40 μmol·L^-1 myricetin group was significantly increased (P<0.05) and the proportion of GL261 cells in S phase was decreased(P<0.05) compared with control group.The Hoechst 33342 staining results showed that the apoptotic bodies appeared in myricetin groups.The early and late apoptotic rates of GL261 cells in myricetin groups detected by AnnexinⅤ/PI double staining were increased compared with control group(P<0.05). Compared with control group,the number of migrated GL261 cells in 5,10,and 20 μmol·L^-1 myricetin groups was decreased in Traswell chamber assay (P<0.05).Compared with control group,the number of GL261 cells entering the lower chamber in myricetin groups was decreased significantly(P<0.05);the number of GL261 cells passing the Matrigel was gradully decreased with the increasing of the concentrations of myricetin.Compared with control group,the expression levels of MMP-2 and MMP-9 mRNA in the GL261 cells in 5 and 10 μmol·L^-1 myricetin groups were decreased(P<0.05). Conclusion:Myricetin could inhibit the proliferation of GL261 cells and induce the apoptosis. Meanwhile, it could also inhibit the migration and invasion of GL261 cells.

Objective:To study the effects of room curing polymethyl methacrylate(PMMA)with nano-silver on the liver tissue and DNA damage of hepatic cells in the mice, and to evaluate the hepatotoxicity of the nanocomposites. Methods:A total of 40 male Kunming mice aged 6-8 weeks were divided into room curing PMMA with nano-silver groups(PMMA-NM groups,72 h extract liquid, 1/2 72 h extract liquid, 1/4 72 h extract liquid),room curing PMMA groups(PMMA groups, 72 h extract liquid, 1/2 72 h extract liquid, 1/4 72 h extract liquid), negative control group, and positive control group(n=5).All mice were treated by gavage with test compounds at the beginning of experiment and 24 h and 45 h after experiment except the positive control agent, the mice in positive control group were treated with ethylmethylsulfone at 24 and 45 h after experiment,and the mice in negative control group were administrated with the same volume of normal saline by gavage. The eyeball blood of mice was collected for biochemistry analysis of liver 3 h after the final administration. The liver tissue was completely removed immediately after the mice were sacrificed and the organ coefficient was calculated. The percentage of tail DNA(%tail DNA), tail length(TL) and Olive tail moment(OTM) were detected by the in vivo comet assay,and the images were analyzed with analysis software of comet assay. Results:Compared with negative control group, the liver coefficient, the alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels in liver tissue of the mice in various experimental groups had no significant differences(P>0.05); the% tail DNA,TL and OTM in PMMA-NM groups and PMMA groups had no significant differeces (P>0.05).The% tail DNA,TL and OTM of the mice in positive control group were higher than those in negative control group(t=-40.911, P<0.05;t=-16.620, P<0.05;t=32.943, P<0.05). There were no significant differences of the% tail DNA, TL and OTM of the mice between PMMA-NM groups and PMMA groups at the same concentration (P>0.05).There were no significant differences of the%tail DNA, TL and OTM between PMMA-NM groups at different concentrations of extrat liquid(P>0.05). Conclusion:Room curing PMMA with nano-silver doesn't have adverse effects on the liver and its function in the mice, and the results of comet assay indicates that the material has no genotoxicity, showing that the composite has good biocompatibility.

Objective:To detect the optimal fusing temperature, the fusing time and the best filler ratio of α-Si₃N₄ and SP₁SiO₂, and to clarify their influence in the properities of dental resin composites. Methods:The α-Si₃N₄ crystalline were mixed with SP₁SiO₂ particle at the ratio of 5:1(Wt%), and then were sintered under 500℃, 650℃, 800℃, 950℃ and 1 100℃ at a rise rate of 250·h^-1 and maintained for 10 min, 30 min and 3 h, respectively(used as α-Si₃N₄-SP₁SiO₂ groups).SP₁SiO₂ particle, α-Si₃N₄ crystalline, mixed and non-fused α-Si₃N₄ and SP₁SiO₂(mixed) groups were set up, and two commercially available resin composites were selected and used as control groups. And they were fully mixed with the resin matrix in 60%(Wt%) after the treatment of cyclohexane solution to make the samples. The flexural strength of specimen was tested and the morphology of section under SEM was analyzed. Then α-Si₃N₄ was mixed with SP₁SiO₂ in a ratio of 2:1. It was fused under the optimal fusing conditions. After the treatment of cyclohexane solution, it was mixed and polymerized with the resin matrix in the proportions of 20%, 40%, 60%, 70% and 75%(Wt%), and two kinds of commercially available resin composites were selected and used as control groups. The flexural strength of specimens was tested and the morphology of section under SEM was analyzed. Results:The maximum flexural strength value in α-Si₃N₄-SP₁SiO₂ groups was at 800℃ and 30 min(P>0.05);the flexural strength value was significantly higher than those in SP₁SiO₂ group, α-Si₃N₄ crystalline group, mixed group and two control groups(P<0.05), and the morphology of section SEM was consistent with the mechanical properties. The flexural strengths of resin composites were increased gradually with the increasing of filler ratios of α-Si₃N₄ and SP₁SiO₂ fusion from 20% to 60%(P<0.05); the flexural strength values of resin composites with the proportion of 60%-70% were not increased significantly, the flexural strength values of resin composites with the proportion of 70%-75% were decreased, and the flexural strength values of the resin composites with the proportion of 60% and 70% were significantly higher than those of resin composites with the proportions of 20%, 40%, 75% and control groups; the morphology of section under SEM was consistent with the mechanical properties. Conclusion:The optimum fusing condition for α-Si₃N₄ and SP₁SiO₂ is 800℃ for 30 min,and the best filler ratio of α-Si₃N₄ and SP₁SiO₂ is 70%.

Objective:To investigate the protective effect of salvianolic acid B on the cardiomyocytes of the suckling rats with oxidative stress injury, and to clarify its mechanism. Methods:The cardiomyocytes of Wistar suckling ratswere primary cultured in vitro and randomly divided into normal control group, model group, low dose of salvianolic acid B group, and high dose of salvianolic acid B group. Except normal control group, the suckling rat cardiomyocytes were induced to oxidative stress injury by being exposed to H₂O₂ at the dose 100 μmoL·L^-1; 20 and 40 μmol·L^-1 salvianolic acid B were separatly added into low and high doses of salvianolic acid B groups. After co-culturing for 4 h, the morphological changes of cardiomyocytes were observed by light microscope;the survival rate of cardiomyocytes was examined using MTT. The levels of reductive glutathione (GSH), the activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) in the cardiomyocytes were detected with spectrophotometry. The levels of lactate dehydrogenase(LDH) and creatine kinase (CK) in culture medium were determined by ELISA. Results:Compared with normal control group, the cardiomyocytes in model group partially suspended in the culture medium, the number of adherent cells was decreased, the pseudopod contraction and the volume were decreased, and the self-discipline throb of cardiomyocytes was slow. Compared with model group,the number of cardiomyocytes in 20 and 40 μmol·L^-1 salvianolic acid B groups were increased significantly, the pseudopod contraction was reduced, and self-discipline throb was enhanced. Compared with normal control group, the survival rate of cardiomyocytes in model group was decreased(P<0.01), the level of GSH in cardiomyocytes was decreased(P<0.01), the level of MDA was increased (P<0.01),the activity of SOD was decreased(P<0.01), and the levels of CK and LDH in culture medium were increased (P<0.05 or P<0.01). Compared with model group, the survival rates of cardiomyocytes in 20 and 40 μmol·L^-1 salvianolic acid B groups were increased(P<0.05 or P<0.01), the levels of GSH and the activities of SOD were increased(P<0.05 or P<0.01), the levels of MDA were decreased (P<0.05 or P<0.01),and the CK and LDH levels in culture medium were decreased (P<0.05 or P<0.01). Conclusion:Salvianolic acid B can protect the cardiomyocytes against oxidative stress injury, and its mechanism may be associated with the inhibition of lipid peroxidatic reaction and the enhancement of antioxidant ability of cardiomyocytes.

Objective:To investigate the expressions of phosphorylated extracellular signal-regulated kinase (pERK) in the hippocampus of the mice of embryonic(E)18 d and postnatal(P)1,3,7,14,21 and 28 d,and to discuss its possible role during the hippocampus development of the mice. Methods:The hippocampus tissues of the mice of E 18 d,P 1 d,P 3 d,P 7 d,P 14 d,P 21 d and P 28 d were obtained.Each time point was used as observation group. Immunohistochemistry,Western blotting and image analysis technique were used to analyze the expression levels of pERK protein in hippocampus tissue at each time point. Results:The immunohistochemistry results revealed that pERK protein mainly expressed in the nucleus of the nerve cells in the granular layer of dentate gyrus and pyramidal layer of Ammon's horn in hippocampus. In the mice of E 18 d, the expression of pERK in DG and CA regions of hippocampus were slightly stained and arranged sparsely, and the expression level was low;and the pERK was deeply stained and the expression levels of pERK were increased gradually with the development of the hippocampus from E 18d to P 7 d;compared with other six time points, the expression level of pERK of the mice of P 7 d was the highest(F=34.537, P<0.01).From P 14 d to P 21 d,the pERK was slightly stained and the density was decreased;the expression levels of pERK were decreased (F=28.754,P<0.01),and then remained stably on P 28 d(F=6.075, P>0.05). The Western blotting results showed that pERK expressed at each time point,from E18 d to P7 d,the expression levels of pERK were increased gradually; compared with other six time points,the expression level of pERK on P 7 d was the highest (F=33.856, P<0.01).The expression levels of pERK were decreased from P 14 d to P 21 d(F=22.627,P<0.01),and then remained stably on P 28 d(F=7.421,P>0.05). Conclusion:The hippocampus development is increased significantly at the early stage of the mice(E18-P 7d),and the expression levels of pERK are increased gradually in the meanwhile;the expression level peaks on P 7 d, and then it is gradually decreased until a stable level. pERK may take part in the cell proliferation in the development of hippocampus of the mice.

Objective:To explore the effects of miR-124a on the expression levels of tumor necrosis factor alpha (TNF-α)and interleukin-6(IL-6) and cell cycle in the macrophage J774.1 cells of the rheumatoid arthritis(RA) model mice, and to elucidate the mechanism of miR-124a in the pathogenesis of RA. Methods:The J774.1 cells in logarithmic growth phage were obtained and uniformly inoculated in the petri dish with 1×10⁶ mL^-1,and there were 3 multiple holes in each group;transfection was carried out when the fusion degree of the cells reached 60%.The cells were divided into blank control group(without any treatment), empty vector group(transfected with adenovirus negative fluid)and miR-124a overexpression group(transfected with miR-124a adenovirus vector).The expression levels of TNF-α and IL-6 in supernatant of the cells in three groups were detected by ELISA 48 h after transfection. RT-PCR was used to detect the relative expression levels of TNF-α and IL-6 mRNA in J774.1 cells 48 h after transfection and the changes of cell cycle were detected using flow cytometry. Results:The expression levels of TNF-α and IL-6 in supernatant of the cells in miR-124a overexpression group were lower than those in blank control group and empty vector group 48 h after transfection(P=0.038,P=0.042;P=0.043,P=0.044).The expression levels of TNF-α mRNA and IL-6 mRNA in miR-124a group were significantly lower than those in blank control group and empty vector group(P=0.001,P=0.002;P=0.001,P=0.003).The pecentage of cells in G₁ phase in miR-124a overexpression group was higher than those in blank control group and empty vector group(P<0.01);the percentages of cells in S phase and G₂ phase were lower than those in blank control group and empty vector group(P<0.01). Conclusion:MiR-124a can decrease the expression levels of inflammatory cytokines after transfecting the macrophages J774.1 cells,and inhibit the proliferation of cells.MiR-124a is a potential therapeutic target for the treatment of RA.

Objective:To investigate the effects of silymarin on the proliferation, migration and invasion of human gastric cancer cell line SGC 7901 and its role in promoting apoptosis,and to clarify their possible mechanisms. Methods:The human gastric cancer SGC 7901 cells in logarithmic growth phase were selected and divided into control group (without silymarin) and different concentrations(15,30, and 60 mg·L^-1)of silymarin groups.Inverted microscope was used to observe the morphology of cells.MTT method wasused to detect the cell cycle and the apoptotic rates of cells in various groups.The inhibitory rates of proliferation of cells in various groups were detected by flow cytametry. Scratch test and Transwell chamber assay were used to detect the cell migration and invasion abilities. Results:After the SGC 7901 cells were treated with silymarin for 24 h, compared with control group, the adherent densities of the cells in 30 and 60 mg·L^-1 silymarin groups were smaller,the cell shape was irregular and the volume was smaller, resulting in a large number of cell debris.Compared with control group, the inhibitory rates of cells in different concentrations of silymarin groups were increased significantly(P<0.05); the percentages of cells in G₁ phase and the apoptosis rates of SGC 7901 cells were increased significantly(P<0.05);the migration distances were decreased(P<0.05) and the number of transmembrane cells was decreased significantly(P<0.05). Conclusion:Silymarin can inhibit the proliferation of human gastric cancer SGC 7901 cells by inducing the G₁ phase cell arrest and early apoptosis.

Objective:To investigate the protective effect of Qijudihuang Pill on the retinal structures of the glaucoma model rats, and to clarify its mechanism. Methods:Fifty SD rats were randomly divided into blank control group, model group,and low, medium, and high doses of Qijudihuang Pill groups(n=10). The chronic glaucoma rat models were established by using cauterization on episcleral veins in each group except blank control group. The rats in low, medium, and high doses of Qijudihuang Pill groups were given 1,2, and 4 g·kg^-1 Qijudihuang Pill by gavage, while the rats in blank control group and model group were given the same volume of saline, 1 time per day. After 12 weeks of administration, the morphological changes in ocular tissue were analyzed by HE staining. The apoptosis of retinal ganglion cells was detected by TUNEL staining. The expression levels of Bcl-2, Bax,and caspase-3 mRNA in the retina tissue of the rats in various groups were determined by RT-PCR.The expression levels of Bcl-2, Bax, and caspase-3 proteins in the retina tissue of the rats in various groups were determined by immunohistochemistry. Results:Compared with blank control group, the retinal nerve fibers in model group were irregularly arranged, the interfibrillar substance was edematous, the cell layer was vacuolated, and the number of retinal ganglion cells was reduced significantly.Compared with model group, the number of retinal ganglion cells of the rats in high dose of Qijudihuang Pill group was increased; the retinal nerve fiber cells in the eyes of the rats in low dose of Qijudihuang Pill group and medium dose of Qijudihuang Pill group were arranged neatly, and the morphology of ganglion cells were normal.Compared with model group, the apoptotic indexes of retinal ganglion cells in low, medium, and high doses of Qijudihuang Pill groups were decreased significantly(P<0.05); the mRNA and protein expression levels of Bcl-2 were increased significantly(P<0.05), while the mRNA and protein expression levels of Bax and caspase-3 were decreased significantly(P<0.05). Conclusion:Qijudihuang Pill can protect the retina of glaucoma rats by regulating the mRNA and protein expressions of Bcl-2, Bax, and caspase-3 and inhibiting the apoptosis of retinal ganglion cells.

Objective:To explore the changes of levels of endogenous N-acetyl-seryl-aspartyl-lysylproline (AcSDKP) and its regulatory factors in liver tissue of rats with liver fibrosis induced by bile duct ligation, and to elucidate the effect of endogenous AcSDKP in the pathologic process of liver fibrosis. Methods:Forty-five healthy male rats were randomly divided into model group (the liver fibrosis rat models were set up by bile duct ligation), blocking group (the liver fibrosis rat models were treated with angiotensin-converting enzyme inhibitor before) and control group (the rats received laparotomy but not bile duct ligation); each group had 15 rats. The serum and liver tissue of the rats in various groups were gained at the 1st week, 2nd week and 4th week. The differences in the indexes of liver function, the liver fibrosis degrees,and the AcSDKP levels in liver tissue of the rats in various groups were analyzed. The levels of thymosin beta 4 (Tβ4), prolyl oligopeptidase (POP) and angiotensin converting enzyme (ACE) in liver tissue of the rats in various groups were detected by Western blotting. Results:The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST) and total bilirubin(TB) of the rats in model group were higher than those in other two groups from the 1st week (P<0.05), and there were no differences between blocking group and control group (P>0.05). The ALB and the AcSDKP level in liver tissue of the rats in model group were lower than those in other two groups from the 2nd week (P<0.05), and there were no differences between blocking group and control group (P>0.05).The levels procollagen type Ⅲ(PCⅢ), collagen type Ⅳ(CⅣ) and hyaluronic acid(HA) in three groups had no differences at the 1st week (P>0.05). The levels of PCⅢ, CⅣ, and HA of the rats in model group were higher than those in other two groups from the 2nd week (P<0.05), and there were no differences between blocking group and control group (P>0.05). The levels of Tβ4 and ACE in liver tissue of the rats in model group were higher than those in other two groups from the 2nd week (P<0.05), and the level of POP of model group were lower than those in other two groups from the 2nd week (P<0.05), but there were no differences between blocking group and control group (P>0.05). The liver tissue of the rats in model group was basically normal at the 1st week, showed the focal necrosis at the 2nd week, and showed the flake necrosis at the 4th week. The liver tissue of the rats in blocking group and control group were basically normal at all time points. Conclusion:The endogenous AcSDKP level in the liver fibrosis rats induced by bile duct ligation is down-regulated through the axis of Tβ4-POP-AcsDKP.

Objective:To investigate the effects of Chaishao Rupi Granules on the levels of serum hormones and the expression levels of vascular endothelial growth factor(VEGF) and basic fibroblast growth factor (bFGF) proteins in breast tissue of the model with mammary gland hyperplasia rats, and to clarify its therapeutic mechanism. Methods:Sixty female SD rats were selected and randomly divided into control group, model group, tamoxifen group (4 mg·kg^-1),and low(0.45 g·kg^-1), medium (0.90 g·kg^-1) and high (1.80 g·kg^-1) doses of Chaishao Rupi Granules groups. The rats in control group were intraperitoneally injected with normal saline (0.05 mL), and fed with normal diet;the rats in other groups were intraperitoneally injected with 0.5 mg·kg^-1 estradiol benzoate for 25 d, then they were intraperitoneally injectcel with progesterone injection 5 mg·kg^-1 for 5 d to establish the rat models of mammary gland hyperplasia, continuous administration for 8 weeks. The diameters and heights of the second nipples of the rats were measured, the blood of abdominal aorta was taken and the levels of estradiol (E2), progesterone (P), luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (PRL), gonadotropin-releasing hormone (GnRH) and the densities of VEGF and bFGF were detected with ELISA. Immunohistochemistry was used to detect the expressions of VEGF and bFGF in breast tissue of the rats in various groups. Results:Compared with model group, the acinar shapes in low, medium, and high doses of Chaishao Rupi Granules groups were regular, the secretion was reduced, and the degrees of cell proliferation were reduced.Compared with model group, the heights and diameters of nipples of the rats in low, medium, and high doses of Chaishao Rupi Granules groups were significantly decreased (P<0.05),the levels of E2, LH, PRL, GnRH, VEGF and bFGF in serum were significantly decreased (P<0.05), and the levels of serum P and FSH were significantly increased (P<0.05).Compared with model group, the expression levels of VRGF and bFGF in breast tissue of the rats in low, medium,and high doses of Chashao Rupi Granules groups were significantly decreased (P<0.05). Conclusion:Chaishao Rupi Granules have obvious therapeutic effect in the mammary hyperplasia model rats, and its mechanism is to regulate the serum levels of multiple hormones and the expression levels of VEGF and bFGF proteins in brease tissue of the rats to treat the mammary gland hyperplasia.

Objective:To explore the differences in the levels of 25 hydroxy vitamin D3(vitamin D3) of the asthmatic children with different control levels, to analyze the relationships between the vitamin D3 level and the pulmonary function indexes, and to clarify the effects of vitamin D3 level on the asthma control and the pulmonary function. Methods:A total of 64 asthmatic patients who aged from 6 to 14 years old were selected as asttma group, and then according to the control level, asthma groups were divided into well-controlled group, partially controlled group and uncontrolled group.Eighteen healthy children who aged from 6 to 14 years old were selected as control group.The general materials of subjects in gwo groups were collected. The levels of serum vitamin D3 of the subjects in two groups were detected by Roche electrochemiluminescence, and the pulmonary function indexes(FEV1,FEV1% pred, FEV1/FVC%, and FVC)were measured by spirometry. The levels serum of vitamin D3 of the subjects in asthma and control groups were analyzed by Wilcoxon rank sum test, the differences of vitamin D3 levels among different control level groups were analyzed by Kruskal-Wallis test, and the relationships between the vitamin D3 level and the pulmonary function indexes were analyzed by Spearman correlation analysis. Results:The serum vitamin D3 level of the children in asthma group was significantly lower than that in control group (P<0.05). The level of serum vitamin D3 of the children in well-controlled group was higher than that in uncontrolled group (P<0.05), but the level of serum vitamin D3 of the children in partially controlled group had no differences neither with well-controlled group nor with uncontrolled group (P>0.05). The percentage of children with vitamin D3 deficiency in uncontrolled group was significantly higher than that in well-controlled group (P<0.05). The vitamin D3 level was positively correlated with FEV1 and FEV1/FVC%(r=0.651,P<0.01;r=0.642,P<0.01). Conclusion:The level of serum vitamin D3 of the asthmatic children affects the control level of asthma and pulmonary function, so vitamin D3 inefficiency or deficiency should be paid attention in the treatment of asthma. Vitamin D3 supplementation is expected to be an important means of adjuvant treatment for asthma.

Objective:To observe the clinical application of dexmedetomidine(Dex) combined with non-invasive positive pressure ventilation(NIPPV) inthe patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) complicated with pulmonary encephalopathy, and to clarify its effectiveness Methods:A total of 80 patients with AECOPD and pulmonary encephalopathy were collected and divided into control group and Dex group according to the random digit table(n=40). The patients in control group were treated with conventional therapy, including antibiotics, phlegm, spasmolysis and NIPPV and so on. The patients in Dex group were treated with Dex based on conventional therapy. The heart rate(HR), blood pressure, arterial partial pressure of oxygen(PaO₂), arterial partial pressure of carbondioxide, the rate of endotracheal intubation,the effective rate of NIPPV and the compliance of NIPPV of the patients in two groups were analyzed before and after treatment.The expectoration capacity,delirium and anxiety of the patients in two groups were evaluated. Results:After conventional treatment, the HR, blood pressure and PaCO₂ in two groups were lower than before treatment, and the PaO₂ was higher than before treatment; the degrees of changes of the above indicators in Dex group were more obvious, the differences were statistically significant (P<0.05). At the same time, the intubation rate of the patients in Dex group was lower than that in control group(P<0.05), and the effective rate of NIPPV was higher than that in control group(P<0.05),and the compliance was increased(P<0.05). The ventilatory function, cough ability, expectoration frequency and expectoration volume of the patients in Dex group were not significantly different from those in control group (P>0.05). The incidene rate of delirium and anxiety score of the patients in Dex group were lower than those in control group (P<0.05). Conclusion:Dex combined with NIPPV is a safe and effective method in the treatment of the patients with AECOPD and pulmonary encephalopathy without obvious adverse effects.

Objective:To investigate the expression levels of tumor markers squamous cell carcinoma antigen (SCC-Ag), carcinoembryonic antigen(CEA), cytokeratin 19 fragment(CYFRA21-1)and D-dimer(D-D) of the patients with non-small cell lung cancer(NSCLC), and to elucidate its clinical value in the early diagnosis of NSCLC. Methods:A total of 200 patients with NSCLC(NSCLC group), 198 patients with benign lung disease(benign lung disease group) and 196 healthy subjects(control group) were selected and the levels of tumor markers and D-D were detected and compared between groups. The receiver operator characteristic curve (ROC) was drawn to analyze the diagnostic values of different tumor markers, D-D, and joint indicators in the patients with NSCLC. Results:The levels of tumor markers and D-D of the patients in NSCLC group were significantly higher than those in benign lung disease group and control group (P<0.01). The level of CEA in lung adenocarcinoma was higher than that in lung squamous cell carcinoma (P=0.005). The levels of SCC-Ag and CYFRA21-1 in lung squamous cell carcinoma were higher than those in lung adenocarcinoma (P=0.008, P=0.004). The levels of tumor markers CEA, CYFRA21-1 and D-D of the patients with stage Ⅲ and Ⅳ NSCLC were significantly higher than those of the patients with stage Ⅰ and Ⅱ NSCLC (P<0.05). The ROC curves showed that area under curve(AUC)of SCC-Ag was higher than other single indexes(AUC=0.805);the sensitivity and specificity were 85.42% and 64.21%, respectively. The diagnostic efficacy of the joint indicators was better than that of each single index (AUC=0.933);the sensitivity and specificity were 86.46% and 88.42%, respectively. Conclusion:Combined detection of tumor markers SCC-Ag, CEA, CYFRA21-1 and D-D can significantly increase the sensitivity and specificity of early diagnosis of NSCLC and has important significance in the early diagnosis of NSCLC.

Objective:To study the effect of cellular immune in the pathogenesis of chronic urticaria, and to clarify the immunological mechanism of chronic urticaria. Methods:A total of 68 patients with chronic urticaria and 70 normal controls were selected as the subjects.The patients with chronic urticaria were divided into four different subgroups according to the different complications; there were 11 patients in autoimmune disease group (3 case of rheumatism, 2 case of lupus erythematosus, 4 cases of thyroid diseases, 2 cases vitiligo), 13 patients in Helicobacter pylori infection group, 10 patients in hepatitis B and hepatitis C group, and 6 patients in tumor group. The expressions of T cell surface markers CD3⁺, CD4⁺, and CD8⁺ in the peripheral blood were detected with flow cytometry. Furthermore,the T cell subsets in the peripheral blood of the patients with chronic urticaria with different complications were detected by flow cytometry. Results:Compared with control group, the ratio of CD4⁺ T cells of the patients in chronic urticaria group was significantly decreased (P<0.05), the ratio of CD8⁺ T cells was increased(P<0.05),and the ratio of CD4⁺ T cells/CD8⁺ T cells was decreased (P<0.05);but the ratio of CD3⁺ T cells had no changes (P>0.05).Compared with the patients with the disease duration less than 12 months, the ratio of CD4⁺ T cells of the patients with the disease duration more than 12 months was significantly decreased (P<0.05), the ratio of CD8⁺ T cells was increased(P<0.05),and the ratio of CD4⁺/CD8⁺ was decreased (P<0.05); but the ratio of CD3⁺ T had no changes (P>0.05).Compared with the patients without complications, the ratios of CD4⁺T cells and CD3⁺ T cells of the patients with complications were decreased (P<0.05), the ratio of CD8⁺ T cells was increased(P<0.05),and the ratio of CD4⁺/CD8⁺ was decreased (P<0.05). Conclusion:The unbalance of cellular immune exists in the patients with chronic urticaria, especially the decreasing of the ratio of CD4⁺ T cells in peripheral blood of the patients might be one of the mechanisms of pathogenesis of chronic urticaria.

Objective:To investigate the efficacies of pulsed continuous cardiac output (PICCO) and central venous pressure (CVP) monitoring in the treatment of the patients with sepsis shock complicated with cardiac insufficiency, and to clarify the clinical significance of PICCO and CVP monitoring. Methods:A total of 137 cases of septic shock combined with cardiac insufficiency patients were randomly divided into CVP group (n=68) and PICCO group (n=69). The patients in CVP group were treated with CVP monitoring to guide therapy, and the patients in PICCO group were treated with PICCO monitoring to guide therapy.The levels of lactic acid and B brain natriuretic peptide (BNP), central venous oxygen saturation (ScvO₂), heart rate (HR), mean arterial pressure (MAP) and CVP of the patients in two groups were detected before treatment and 6, 24, 48 h after treatment, respectively; the hemodynamic parameters of the patients of PICCO group were recorded; the vasoactive drug use time,the ICU hospitalization time, the incidence rates of multiple organ dysfunction syndrome (MODS), the mechanical ventilation time and the 28 d-mortality rates of the patients were recorded. Results:Compared with CVP group, the levels lactic acid and BNP and HR of the patients in PICCO group at 6, 24, and 48 h after treatment were significantly decreased (P<0.05), and the scavenging rates of lactic acid were significantly increased(P<0.05); the levels of ScvO₂, MAP and CVP were significantly increased (P<0.05). The levels of MAP of the patients in PICCO group were significantly higher than those in CVP group at the same time(P<0.05), after 48 h of treatment, the ScvO₂ levels in PICCO group were significantly higher than those in CVP group at the same time(P<0.05), but there was no significant difference in the CVP levels of the patients between two groups at the same time (P>0.05). There were no significant differences in the incidence rates of MODS and 28 d-mortality rates between two groups(P>0.05); however the use time of vasoactive drugs, the ICU hospitalization time and the mechanical ventilation time of the patients in PICCO group were significantly shorter than those in CVP group (P<0.05). Conclusion:PICCO monitoring guides the treatment of septic shock complicated with cardiac insufficiency better than CVP monitoring, and has positive clinical significance.

Objective:To investigate the influence of high-purity human menopausal gonadotrophin (HP-hMG) added in different periods of antagonist program COH in the pregnancy outcom of in vitro fertilization-embryo transplantation (IVF-ET) of the patients with normal ovarian function reserve,and to clarify the possible mechanism. Methods:A totol of 142 patients aged from 25 to 40 years old with normal ovarian function underwent IVF treatment were selected.According to the timing of HP-hMG addition,they were divided into early addition group (HP-hMG was added on the first day) and middel and latel addition group (HP-hMG was added on the 6th day).The pregnancy outcomes of the patients were analyzed. Results:Compared with early addition group, the estradiol(E2) level on the day of human chorimic gonadotropin(HCG) injection in middle and late addition group was significantly increased (P=0.042) and the progesterone(P) level on the day of HCG injection was significantly decreased(P=0.016), the number of follicles with diameter > 16 mm (P=0.035) and the number of follicles with diameter > 18 mm(P=0.026) were increased.Compared with early addition group,the rate of mature follicles in middle and late addition group was increased (P=0.006),the rate of high quality embroy was increased (P=0.001),and the cumulative pregnancy rate was increased (P=0.040). Conclusion:The addition of HP-hMG at middle and late follicular phase can improve the pregnancy outcome in the patients with normal ovarition function reserve under went antagonist program COH.

Objective:To detect the expressions of transforming growth factor-β1 (TGF-β1) and placental growth factor (PLGF) expressions in the tissue of the patients with different types of gestational trophoblastic diseases,and to clarfy the significances of their expressions in different types of gestational trophoblastic diseases. Methods:A total of 40 cases of hydatidiform mole tissue (hydatidiform mole group) and 26 cases of gestational trophoblastic tumour tissue (GTN group, including 22 cases of invasive hydatidiform mole and 4 cases of choriocarcinoma), and 40 cases of normal early pregnancy villi (normal control group) were collected.Immunohistochemical method was used to detect the positive expression rates of TGF-β1 and PLGF, the differences of TGF-β1 and PLGF expressions between normal group, hydatidiform mole group and GTN group were analyzed, and the associations between them and the clinical high risk factors(age>40 years old, the uterine volume> gestational age, and thecalutein ovarian cyst diameter>6 cm) were analyzed. Results:The positive expression rates of TGF-β1 in hydatidiform mole group(62.5%)and GTN group(30.8%) were lower than that in normal group(87.5%) (P<0.05);the positive expression rates of TGF-β1 in hydatidiform mole group was higher than that in GTN group(P<0.05).The positive expression rates of PLGF in hydatidiform mole group(50.5%) and GTN group(80.8%%) were higher than that in normal control group(22.5%%);the positive expression rate of PLGF in hydatidiform mole group was lower than that in GTN group (P<0.05).The positive expression rates of TGF-β1 of the patients in hydatidiform mole group with the age>40 years old, the uterine volume>the gestational age,and the ovarian luteinized cyst diameter>6 cm were significantly decreased (P<0.05) and the positive expression rates of PLGF were significantly increased (P<0.05) compared with normal control group.The positive expression rate of TGF-β1 of the patients in GTN group with the high risk factors was significantly decreased compared with those without the high risk factors(P<0.05) and the positive expression rate of PLGF was significantly increased (P<0.05).In hydatidiform mole group and GTN group, there were moderately negative correlations between the positive expression rates of TGF-β1 and PLGF (r=-0.585,P<0.05;r=-0.479, P<0.05). Conclusion:The expression levels of TGF-β1 in hydatidiform mole and gestational trophoblastic tumor are gradually decreased, and the expression levels of PLGF are gradually increased, which may be related to the different types and prognosis of gestational trophoblastic diseases.

Objective:To analyze the expressions of myeloid differentiation factor 88(MyD88) in cancer tissue and adjacent tissue of the patients with colorectal cancer and to explore the relationships between its expression and the clinicopathological parameters and the prognosis of the patients with colorectal cancer, and to clarify the clinical significance of MyD88 expression. Methods:A total of 90 cases of tumor tissue and adjacent tissue of the patients with colorectal cancer were collected and their clinical pathological materials were analyzed. The expressions of MyD88 in tumor tissue and adjacent tissue of the colorectal cancer patients were detected by immunohistochemistry. The expression levels of MyD88 in different tissues were analyzed by Image-Pro Plus 6.0 software. Kaplan-Meier method was used for survival analysis in the 90 patients with colorectal cancer. Results:The expression level of MyD88 in colorectal cancer tissue was significantly higher than that in adjacent colorectal cancer tissue(P<0.001). The MyD88 expression level had no correlations with the age, gender, tumor size and histologreal differentiation of the patients with colorectal cancer (P<0.05). The MyD88 expression level had correlations with the clinical stage, T stage, M stage and lymph node metastasis(P<0.05). The survival rate of the patients with colorectal cancer with higher expression of MyD88 was significantly lower than that of the patients with colorectal cancer with low expression of MyD88 (P<0.05). Conclusion:MyD88 is highly expressed in colorectal cancer tissue and it is associated with the clinical stage, T stage, M stage, and the situation of lymph node metastasis of the patients with colorectal cancer.

Objective:To study the expressions of AAX14401 in the normal bladder tissueand the human bladder transitional cell carcinoma tissue, and to analyze its function in the occurrence and development of bladder cancer. Methods:Seventy-five cases of specimens of bladder transitional cell carcinoma were collected and used as experimental group and 12 cases of normal bladder mucosa specimens were selected as control group. The positive expression rates of glutathione S-transferase pi-1(GSTP1) and AAX14401 were detected by immunohistochemistry. The differences of positive expression rates of GSTP1 and AAX14401 proteins between bladder cancer tissue and normal bladder tissue were analyzed. Results:The positive expression rate of GSTP1 protein in normal bladder tissue was 58.3%, while it was 90.7% in bladder cancer tissue, and the difference between them was significant (P<0.01). The positive expression rate of AAX14401 protein in normal bladder tissue was 0%, while it was 2.7% in blandder cancer tissue, and there was no significant difference between them(P=0.596). There was no one with positive AAX14401 expression and positive GSTP1 expression in bladder cancer tissue at the same time. There were 2 cases with positive AAX14401 expression in bladder cancer tissue with negative GSTP1 expression; there was a negative correlation between the positive expressions of AAX14401 and GSTP1 proteins in bladder cancer (r=-0.274, P=0.018). Conclusion:The expression of AAX14401 protein may promote the incidence and progression of bladder cancer.

Objective:To observe the clinical efficacy and safety of tacrolimus(TAC) combined with low dose of corticosteroids in the treatment of the children with refractory nephrotic syndrome(RNS),and to investigate the effect of diltiazem on the blood concentration of TAC. Methods:A total of 16 hospitalized children with refractory nephrotic syndrome(RNS)were selected.All patients were treated with TAC(0.10-0.15 mg·kg^-1·d^-1)combined with low dose of corticosteroids(l.0-2.0 mg·kg^-1·d^-1).The concentrations of TAC of the children during the treatment process were monitored.The 24 h urine protein quantitative(UP),plasma albumin (Alb),creatinine and other 6 indexes were recorded before treatment and 2,4,8,12 and 24 weeks after treatment. Results:In 16 cases,9 cases were completely relieved,5 cases were partially relieved,and 2 cases were invalid;the total efficiency was 94%.The 24 h UP of 16 children at 24 weeks after treatment was significantly decreased compared with before treatment(Z=-3.516,P<0.01);the plasma Alb level of the children was significantly increased compared with before treatment(Z=-3.516,P<0.01).The total cholesterol and triglyceride levels at 2 weeks after treatment were decreased significantly compared with before treatment(Z=-2.223,P<0.05;Z=-3.464,P<0.01).Eight patients had low blood concentration of TAC,and it was increased to an effective concentration after treated with diltiazem.In the course of treatment,creatinine,urea nitrogen and other indexes of the patients were fluctuated in a normal range. Conclusion:TAC combined with low dose of corticosteroids for treatment of RNS is safe and effective,and diltiazem can effectively increase the blood concentration of TAC.

Objective:To compare the curative effects and adverse reactions of fractional carbondioxide laser combined with tacrolimus ointment and tacrolimus ointment alone in the treatment of localized vitiligo,and to evaluate their safeties. Methods:The clinical materials of 98 patients with localized vitiligo were retrospectively analyzed.The patients were divided into two groups according to their wishes.The patients in control group (47 cases) were treated with tacrolimus ointment every morning and evening only, and the patients in observation group (51 cases) were treated with fractional carbondioxide laser combined with tacrolimus ointment and the laser was used once every 4 weeks. The treatment lasted for 12 weeks. Compound Glycyrrhizin Tablets were used according to the conditions (3 pieces each time, 3 times a day, a total of 12 weeks). The follow-up period was once every 2 weeks. The clinical efficacy and adverse reactions of the patients in two groups were assessed in the 12th weeks after treatment. Results:The onset time of treatment of the patients in observation group was shorter than that in control group(t=13.82,P=0.001), and the total effective rate of the patients in observation group was higher than that in control group(χ²=4.72,P<0.05). The effective rates of the faciocervical lesion of the patients in observation group and control group were higher than those of the trunk lesion(χ²=23.78, P<0.05;χ²=12.79, P<0.05). The effective rate of the faciocervical lesion of the patients in observation group was higher than that in control group (χ²=5.75,P<0.05); there were no serious adverse reactions in all patients. Conclusion:The combination of fractional carbondioxide laser and tacrolimus ointment in the treatment of localized vitiligo has the advantages of quick effect, good efficacy and high safety, and is worthy of clinical application.

Objective:To explore the clinical diagnosis and treatment of huge thrombus in ascending aorta, and to clarify its treatment methods. Methods:The clinical materials of one patient with ascending aortic thrombus were retrospectively analyzed and the diagnosis and treatment were summerized, and the relevent literatures were reviewed. Results:The patient was male, and admitted to hospital due to space-occupying lesion in the ascending aorta in routine examination. The results of aortic CTA reported that there was a 22 mm×22 mm×45 mm space-occupying lesion in the ascending aorta. The patient was successfully treated with thrombectomy and ascending aorta replacement in order to prevent the systemic embolization.The histopathological examination revealed that the lesion was thrombus. Conclusion:Aortic CTA is a useful examination for the patients with thrombus in ascending aorta. Resection of thrombus and implantation of vascular graft can reduce the risk of recurrent stroke effectively.

Objective:To explore the radionuclide distribution characteristics of ^99mTc-MDP bone imaging of bone tumors or tumor-like lesions in extremities, to clarify the application value of radionuclide distribution characteristics of ^99mTc-MDP bone imaging of bone tumor and tumor-like lesions with different pathological types in the differential diagnosis, and to provide the clues for clinical diagnosis. Methods:A total of 76 patients with abnormal radionuclide distribution characterics of exetremities showed in ^99mTc-MDP bone imaging were selected. The results of ^99mTc-MDP bone imaging were compared with the results of the final clinica diagnosis.The differences in the age, lesion location, radienuclide distribution characteristics of the patients with different pathological types were analyzed. Semi quantitative method (T/N) was used to detect the degree of radioactivity and the differences in T/N between benign and malignant lesions groups. Results:① Among 76 patients with bone tumors and tumor-like lesions,the pathological benign findings were in 24 cases and the malignant findings were in 52 cases.②The age of the patients with giant cell tumor of bone around knee joint was (42.1 ±17.4)years old,and the age of the patients with fibrous dysplasia in proximal femur was (48.0±17.1) years old. In malignant lesions,the age of the patients with metastatic carcinoma located in femoral shaft was (64.0 ±14.2)years old; the age of the patients with osteosarcoma around epiphysis end was (30.3±15.3) years old; the age of the patient with Ewing's sarcoma located in femur was (49.2 ±4.7) years old; the age of patients with fibrosarcoma in long bone was (39.5 ±17.2)years old; the age of patients with chondrosarcoma occurred in long bone was (63.0±14.8)years old. ③The radionuclide distribution characteristics of giant cell tumor, osteosarcoma, Ewing's sarcoma, chondrosarcoma and fibrosarcoma were "wedge" or "lumpy";the radiological defect of giant cell tumor was greater than Ewing's sarcoma and fibrosarcoma, while osteosarcoma and chondrosarcoma rarely had center defects; the radionuclide distribution characteristics of fibrous dysplasia and metastasis were "strip-type",and the metastases involving bilateral cortical bone had the radioactive defect in the central region, but fibrous dysplasia distributed uniformly along with the unilateral cortical bone. ④ The average value of T/N in malignant lesions (3.38±1.95) was higher than that in benign lesions (1.43±0.51)(t=-11.35, P<0.01). Conclusion:The pathological types of lesions can be preliminarily speculated according to the characteristics of ^99mTc-MDP bone imaging of bone tumors and tumor-like lesions of extremities, which can provide a reliable reference for clinical diagnosis and differential diagnosis.

Objective:To explore the correlations between the ultrasound features and the expressions of the molecular biological indicator human epidermal growth factor receptor 2(CerbB-2) and Ki-67 in the patients with breast cancer. Methods:The ultrasonographic characteristics and paraffin section materials of 107 patients with breast cancer confirmed by pathology were collected. The correlations between the ultrasonographic characteristics and the expressions of CerbB-2 and Ki-67 were analyzed. Results:The positive expression rates of CerbB-2 and Ki-67 in breast cancer tissue were 73.83%(79/107) and 81.21%(87/107), respectively;the expressions of CerbB-2 and Ki-67 were positively correlated(r=0.264,P=0.001).There were obvious correlations between the edge, size, shape, orientation, hyper-echoic halo, rear echo attenuation,micro calcification, flow signal 2-3 grade of breast cancer and the expression of CerbB-2 (P<0.05).There were obvious correlations between the aspect ratio >1, hyper-echoic halo, rear echo attenuation,micro calcification, flow signal 2-3 grade of breast cancer and the expression of Ki-67 (P<0.05). The results of multivariate Cox regression analysis showed that CerbB-2 (⧻)(P=0.002), Ki-67 (⧻)(P=0.006), blood flow 2-3 grade (P=0.036), rear echo attenation (P=0.001),and aspect ratio>1 (P=0.002) were the independent prognostic factors of breast cancer. Conclusion:There are significant correlations between some ultrasound features of breast cancer and the positive expressions of CerbB-2 and Ki-67 in the breast cancer tissue.

Objective:To investigate the status of overweight/obesity of the children and adolescents in Yanji City in 2016 and related lifestyles, and to clarify the influencing factors of overweight/obesity. Methods:A census method was used to investigate the heights and weights of 42 132 children and adolescents aged from 6 to 17 years old in urban area of Yanji City, and the body mass index (BMI) was calculated. Using stratified cluster random sampling method and questionnaire survey, a total of 1 523 children and adolescents aged 10-14 years old were enrolled to investigate dietary habits. The standard developed by Chinese Obesity Work Group was used to screen the children and adolescents with overweight and obesity. Using Logistic regression and mediating effect analysis method, the influencing factors of overweight/obesity were screened. Results:In 2016, the total detection rate of overweight and obesity of the children and adolescents aged from 6 to 17 years old in urban area of Yanji City were 16.7% and 18.4%, respectively;the total detection rate of overweight and obesity was 35.0%. The detection rates of overweight and obesity in the boys(18.6%, 22.0%) were higher than those in the girls (14.7%, 14.4%) (P<0.01). The detection rates of overweight in the boys in each age group from 7 to 13 years old were higher than those in the girls at the same age (P< 0.05 or P<0.01), and the detection rates of obesity in the boys in each age group were higher than those in the girls (14.4%) at the same age(P<0.01). The detection rate of overweight at the boys peaked at 12 years old and the obesity detection rates were higher in the boys aged from 6 to 10 years old(23.8%-25.6%); the detection rates of obesity during the ages of 11 to 17 years old were gradually decreased with the age increasing. Eating barbecue food more than 3 times a week (boys:OR=1.767, P=0.010,95%CI=1.148-2.719;girls:OR=2.205,P=0.002,95%CI=1.327-3.664) and dieting to lose weight (boys:OR=2.113, P<0.001,95%CI=1.456-3.065;girls:OR=2.128,P<0.001,95%CI=1.430-3.167) increased the risk of overweight/obesity in the children and adolescents, and eating sweet snacks 3 times or more a week (boys:OR=0.359, P<0.001,95%CI=0.226-0.573;girls:OR=0.324,P<0.001,95%CI=0.186 -0.565) and daily meals on time (boys:OR=0.683, P=0.028,95%CI=0.486-0.960; girls:OR=0.624,P=0.016,95%CI=0.424-0.916) reduced the risk of overweight/obesity. Self-evaluation on body type had a full mediating effect between overweight and obesity and daily meals on time. Conclusion:The detection rate of overweight and obesity among the children and adolescents aged from 6 to 17 years old in Yanji City is at a relatively high level. Dietary habits and dieting are the major factors affecting overweight and obesity.