Observation of Structures in the Processes e

We present measurements of the Born cross sections for the processes e^{+}e^{-}→ωχ_{c1} and ωχ_{c2} at center-of-mass energies sqrt[s] from 4.308 to 4.951 GeV. The measurements are performed with data samples corresponding to an integrated luminosity of 11.0 fb^{-1} collected with the BESIII detector operating at the Beijing Electron Positron Collider storage ring. Assuming the e^{+}e^{-}→ωχ_{c2} signals come from a single resonance, the mass and width are determined to be M=(4413.6±9.0±0.8) MeV/c^{2} and Γ=(110.5±15.0±2.9) MeV, respectively, which is consistent with the parameters of the well-established resonance ψ(4415). In addition, we also use one single resonance to describe the e^{+}e^{-}→ωχ_{c1} line shape and determine the mass and width to be M=(4544.2±18.7±1.7) MeV/c^{2} and Γ=(116.1±33.5±1.7) MeV, respectively. The structure of this line shape, observed for the first time, requires further understanding.

Observation of Significant Flavor-SU(3) Breaking in the Kaon Wave Function at 12

We present cross sections for the reaction e^{+}e^{-}→K_{S}^{0}K_{L}^{0} at center-of-mass energies ranging from 3.51 to 4.95 GeV using data samples collected in the BESIII experiment, corresponding to a total integrated luminosity of 26.5 fb^{-1}. The ratio of neutral-to-charged kaon form factors at large momentum transfers (12

Observation of D

We perform for the first time an amplitude analysis of the decay D^{+}→K_{S}^{0}π^{+}η and report the observation of the decay D^{+}→K_{S}^{0}a_{0}(980)^{+} using 2.93 fb^{-1} of e^{+}e^{-} collision data taken at a center-of-mass energy of 3.773 GeV with the BESIII detector. As the only W-annihilation-free decay among D to a_{0}(980) pseudoscalar, D^{+}→K_{S}^{0}a_{0}(980)^{+} is the ideal decay in extracting the contributions of the W-emission amplitudes involving a_{0}(980) and to study the final-state interactions. The absolute branching fraction of D^{+}→K_{S}^{0}π^{+}η is measured to be (1.27±0.04_{stat}±0.03_{syst})%. The branching fractions of intermediate processes D^{+}→K_{S}^{0}a_{0}(980)^{+} with a_{0}(980)^{+}→π^{+}η and D^{+}→π^{+}K[over ¯]_{0}^{*}(1430)^{0} with K[over ¯]_{0}^{*}(1430)^{0}→K_{S}^{0}η are measured to be (1.33±0.05_{stat}±0.04_{syst})% and (0.14±0.03_{stat}±0.01_{syst})%, respectively.

Observation of a Vector Charmoniumlike State at 4.7 GeV/c

Using data samples with an integrated luminosity of 5.85 fb^{-1} collected at center-of-mass energies from 4.61 to 4.95 GeV with the BESIII detector operating at the BEPCII storage ring, we measure the cross section for the process e^{+}e^{-}→K^{+}K^{-}J/ψ. A new resonance with a mass of M=4708_{-15}^{+17}±21 MeV/c^{2} and a width of Γ=126_{-23}^{+27}±30 MeV is observed in the energy-dependent line shape of the e^{+}e^{-}→K^{+}K^{-}J/ψ cross section with a significance over 5σ. The K^{+}J/ψ system is also investigated to search for charged charmoniumlike states, but no significant Z_{cs}^{+} states are observed. Upper limits on the Born cross sections for e^{+}e^{-}→K^{-}Z_{cs}(3985)^{+}/K^{-}Z_{cs}(4000)^{+}+c.c. with Z_{cs}(3985)^{±}/Z_{cs}(4000)^{±}→K^{±}J/ψ are reported at 90% confidence levels. The ratio of branching fractions {[B(Z_{cs}(3985)^{+}→K^{+}J/ψ)]/B[Z_{cs}(3985)^{+}→(D[over ¯]^{0}D_{s}^{*+}+D[over ¯]^{*0}D_{s}^{+})]} is measured to be less than 0.03 at 90% confidence level.

Relationship between PI3K/mTOR/RhoA pathway-regulated cytoskeletal rearrangements and phagocytic capacity of macrophages.

The objective of this study was to investigate the relationship between PI3K/mTOR/RhoA signaling regulated cytoskeletal rearrangements and phagocytic capacity of macrophages. RAW264.7 macrophages were divided into four groups; blank control, negative control, PI3K-RNAi, and mTOR-RNAi. The cytoskeletal changes in the macrophages were observed. Furthermore, the phagocytic capacity of macrophages against Escherichia coli is reported as mean fluorescence intensity (MFI) and percent phagocytosis. Transfection yielded 82.1 and 81.5% gene-silencing efficiencies against PI3K and mTOR, respectively. The PI3K-RNAi group had lower mRNA and protein expression levels of PI3K, mTOR, and RhoA than the blank and negative control groups (Р<0.01). The mTOR-RNAi group had lower mRNA and protein levels of mTOR and RhoA than the blank and the negative control groups (Р<0.01). Macrophages in the PI3K-RNAi group exhibited stiff and inflexible morphology with short, disorganized filopodia and reduced number of stress fibers. Macrophages in the mTOR-RNAi group displayed pronounced cellular deformations with long, dense filopodia and an increased number of stress fibers. The PI3K-RNAi group exhibited lower MFI and percent phagocytosis than blank and negative control groups, whereas the mTOR-RNAi group displayed higher MFI and percent phagocytosis than the blank and negative controls (Р<0.01). Before and after transfection, the mRNA and protein levels of PI3K were both positively correlated with mTOR and RhoA (Р<0.05), but the mRNA and protein levels of mTOR were negatively correlated with those of RhoA (Р<0.05). Changes in the phagocytic capacity of macrophages were associated with cytoskeletal rearrangements and were regulated by the PI3K/mTOR/RhoA signaling pathway.

[Effects of temperature and relative humidity on the number of outpatients with chronic obstructive pulmonary disease and their interaction effect in Lanzhou, China].

OBJECTIVE: To understand the relationships of daily average temperature and relative humidity with outpatient visit frequency of patients with chronic obstructive pulmonary disease, and whether temperature and relative humidity have a lag effect. METHODS: The effects of daily average temperature, relative humidity, and their interaction in Lanzhou between January 2013 and December 2017 on the outpatient visit frequency of chronic obstructive pulmonary disease patients were analyzed using Poisson generalized linear regression model combined with distributed lag non-linear model. RESULTS: There was a non-linear relationship between the daily average temperature and the outpatient visit frequency of chronic obstructive pulmonary disease patients. Between -12 °C and -8 °C, the outpatient visit frequency increased gradually with the decrease of the daily average temperature, and the outpatient visit frequency of chronic obstructive pulmonary disease patients increased by 11.60% per 1 °C of temperature drop. The daily average relative humidity also presented a non-linear effect on the outpatient visit frequency chronic obstructive pulmonary disease patients. When the daily average relative humidity was in the range of 15%-28%, the outpatient visit frequency increased gradually with the decrease of relative humidity, and the outpatient visit frequency of COPD patients increased by 37.05% for every 1% decrease of relative humidity. A synergistic effect was found between air temperature and relative humidity on chronic obstructive pulmonary disease, that is, under different relative humidity, the effect of air temperature was different. When the daily average relative humidity ≤ 50% and the daily average temperature≤11 °C, the effect of air temperature was the most obvious. For every 1 °C drop in temperature, the daily out-patient visit frequency of the whole population increased by 12.68% (5.62% in males and 7.56% in females; 5.24% in population < 65 years and 14.74% in population ≥ 65 years). When the daily average relative humidity > 50% and the daily average temperature ≤ 11 °C, the daily outpatient visit frequency of the whole population increased by 9.00% for every 1 °C drop in temperature (< 65 years, 7.11%; ≥65 years, 10.93%). When the daily average temperature > 11 °C, the temperature had no effect on the daily outpatient visit frequency of chronic obstructive pulmonary disease patients under different relative humidity. CONCLUSION: The presence of a certain extent of interaction is observed between daily average temperature and relative humidity. Low-temperature and dry environment (relative humidity ≤50%, temperature ≤11 °C) as well as low-temperature and high-humidity environment (relative humidity > 50%, temperature ≤11 °C) can both increase the risk of outpatient visit in chronic obstructive pulmonary disease patients.

Relationship between PI3K/mTOR/RhoA pathway-regulated cytoskeletal rearrangements and phagocytic capacity of macrophages

The objective of this study was to investigate the relationship between PI3K/mTOR/RhoA signaling regulated cytoskeletal rearrangements and phagocytic capacity of macrophages. RAW264.7 macrophages were divided into four groups; blank control, negative control, PI3K-RNAi, and mTOR-RNAi. The cytoskeletal changes in the macrophages were observed. Furthermore, the phagocytic capacity of macrophages against Escherichia coli is reported as mean fluorescence intensity (MFI) and percent phagocytosis. Transfection yielded 82.1 and 81.5% gene-silencing efficiencies against PI3K and mTOR, respectively. The PI3K-RNAi group had lower mRNA and protein expression levels of PI3K, mTOR, and RhoA than the blank and negative control groups (Р<0.01). The mTOR-RNAi group had lower mRNA and protein levels of mTOR and RhoA than the blank and the negative control groups (Р<0.01). Macrophages in the PI3K-RNAi group exhibited stiff and inflexible morphology with short, disorganized filopodia and reduced number of stress fibers. Macrophages in the mTOR-RNAi group displayed pronounced cellular deformations with long, dense filopodia and an increased number of stress fibers. The PI3K-RNAi group exhibited lower MFI and percent phagocytosis than blank and negative control groups, whereas the mTOR-RNAi group displayed higher MFI and percent phagocytosis than the blank and negative controls (Р<0.01). Before and after transfection, the mRNA and protein levels of PI3K were both positively correlated with mTOR and RhoA (Р<0.05), but the mRNA and protein levels of mTOR were negatively correlated with those of RhoA (Р<0.05). Changes in the phagocytic capacity of macrophages were associated with cytoskeletal rearrangements and were regulated by the PI3K/mTOR/RhoA signaling pathway.

[The effect of fine particles on the phagocytosis of alveolar macrophages potentially by Arp2/3 complex in a mouse model of chronic obstructive pulmonary disease].

Objective: To investigate the mechanism of fine particulate matter (PM2.5) on the phagocytosis of alveolar macrophages (AM) in mice with chronic obstructive pulmonary disease (COPD) through actin-related protein (Arp) 2/3 complex. Methods: Forty mice were divided into healthy control(A) group, healthy PM2.5 (B) group, COPD(C) group, and COPD PM2.5(D) group according to the random number table method. A mouse model of COPD was established by cigarette smoke exposure method. PM2.5 (662 µg/m(3)) model was established by continuously inhalation for 90 days in healthy PM2.5 group and COPD PM2.5 group. Flow cytometry was used to detect the ability of AM to phagocytose fluorescein isothiocyanate-labeled E.coli (FITC-E.coli), expressed as mean fluorescence intensity (MFI) and percentage of phagocytic positive cells (phagocytosis percentage); Western blotting was used to detect AM Arp2 and F-actin content, and laser confocal microscopy for AM Arp2 and F-actin and phagocytic FITC-E.coli average optical density and colocalization of Arp2 and F-actin, while scanning electron microscopy was used to observes the morphology of AM after phagocytizing FITC-E.coli. Results: AM phagocytosis: MFI and phagocytosis percentage in the COPD group [4 656±251, (31.9±1.7)%] were lower than the healthy control group [8 657±247, (65.7±1.9)%] (both P<0.01); and healthy PM2.5 group and COPD PM2.5 group [7 653±228, (47.9±1.6)% and 3 660±237, (19.2±1.2)%] were lower than the respective control groups (all P<0.01), and the decrease in the COPD group was more pronounced. AM Arp2, F-actin content: the COPD group (0.51±0.02, 0.46±0.03) were lower than the healthy control group (0.81±0.04, 0.71±0.04, both P<0.01); the healthy PM2.5 group and the COPD PM2.5 group [(0.64±0.03, 0.56±0.04) and (0.29±0.02, 0.26±0.02)] were lower than the respective control groups (all P<0.01), and the decrease in COPD group was more significant. Arp2, F-actin, and phagocytic FITC-E.coli mean optical density values: the COPD group (33.0±2.3, 62.0±0.7, 41.0±0.4) were lower than the healthy control group (141.0±4.2, 145.0±2.9, 189.0±2.6, both P<0.01); the healthy PM2.5 group and the COPD PM2.5 group (127.0±2.8, 124.0±0.7, 154.0±0.9, and 24.0±2.4, 37.0±0.4, 29.0±0.8) were lower than the respective control groups (all P<0.01), and the decrease in the COPD group was more significant. Colocalization of AM Arp2 and F-actin: Montessori colocalization coefficient (MOC) (0.38±0.03) in the COPD group was lower than the healthy control group (0.88±0.03, P<0.01); healthy PM2.5 group and COPD PM2.5 group [(0.58±0.03) and (0.14±0.02)] were lower than the respective control groups (both P<0.01), and the decrease in COPD group was more significant. Morphology of AM phagocytosis of FITC-E.coli: AM in the healthy control group was obviously deformed, and the surface of the cell membrane was slightly wrinkled and high, and the free edge of the micro-pleated fold had a long and dense filamentous pseudopodia extension. The changes of morphology of AM in the COPD group was not obvious, the micro-wrinkles on the surface of the cell membrane were rare, and the filopodia poorly extended or even absent. The AM form of the healthy PM2.5 group changed slightly, mostly irregular circular or elliptical. The micro-wrinkles on the surface of the cell membrane were less and flat, and the filopodia protrudes short and less; the AM form of the COPD PM2.5 group was stiff, and the micro-wrinkles on the surface of the cell membrane were few and flat, no obvious filopodia or protrusions. Correlation analysis: After basal state and PM2.5 intervention, AM Arp2, F-actin content and MOC values of Arp2 and F-actin were positively correlated with MFI. Conclusions: The phagocytic function of AM in COPD mice was low, which was related to the abnormal rearrangement of cytoskeleton involved in Arp2/3 complex and F-actin. It was speculated that PM2.5 might inhibit Arp2/3 complex and F-actin. The cytoskeletal rearrangement of proteins was involved in the aggravation of AM phagocytosis in mice with COPD.

[Effect of actin-related protein 2-3 complex on phagocytosis defect of alveolar macrophages in a mouse model of chronic obstructive pulmonary disease].

Objective: To investigate the role of actin-related protein 2-3 complex (Arp2/3) complex on phagocytosis of alveolar macrophages (AMs) in a mouse model of chronic obstructive pulmonary (COPD). Methods: Forty mice were randomly divided into healthy control group, healthy Arp2/3 complex inhibitor (CK666) group, COPD group and COPD CK666 group with 10 mice in each group. COPD group and COPD CK666 group were established by cigarette smoke exposure, and the control group had no smoke exposure. After 90 days of molding, AMs were isolated from lung tissue of mice in each group. Mean fluorescence intensity (MFI) and the positive percent of AMs engulfing fluorescein isothiocyanate-labeled Escherchina coli (FITC-E.coli) (AM%) were detected by flow cytometry. Western blot was applied to detect protein. Laser scanning confocal microscopy was used to measure the mean optical density of Arp2, F-actin and engulfed FITC-E. coli and quantify the colocalization of Arp2 and F-actin by a Manders' overlap coefficient. Scanning electron microscopy was used to observe the ultrastructure of AM phagocytizing FITC-E.coli. Results: Phagocytosis of AM: MFI and AM% in the COPD group were significantly decreased than those in the healthy control group[(4 702±243), (8 684±234) and (32.21±1.66)%, (65.88±1.77)%, all P<0.01]. MFI and AM% in the COPD CK666 group [(3 597±307), (22.09±1.89)%] and in the healthy CK666 group [(7 446±236), (50.09±1.64)%] were decreased compared to those in their respective control groups (all P<0.01). The expressions of protein of Arp2 and F-actin in the COPD group were significantly decreased than those in the healthy control group (0.508±0.025, 0.813±0.040 and 0.462±0.029, 0.720±0.039) (all P<0.01). The F-actin in the COPD CK666 group (0.265±0.014) and in the healthy CK666 group (0.637±0.032) were significantly decreased compared to those in their respective control groups (all P<0.01). The mean optical density of Arp2, F-actin and FITC-E.coli in the COPD group were significantly decreased compared to those in the healthy group (34.43±0.56, 142.83±1.90 and 61.59±0.70, 145.93±3.05 and 41.49±0.33, 189.17±2.60) (all P<0.01); the mean optical density of F-actin, FITC-E. coli in the COPD CK666 group (37.73±1.04, 28.84±2.95) and in the healthy CK666 group (137.07±1.35, 157.46±1.00) were significantly decreased compared to those in their respective control groups (all P<0.01). The Manders' overlap coefficient of Arp2 and phalloidin' coefficient in the COPD group (0.395±0.014) were significantly decreased than the healthy control group (0.395±0.014 and 0.880±0.002, P<0.01). The Manders' overlap coefficient of Arp2 and phalloidin' coefficient in the COPD CK666 group (0.297±0.006) and in the healthy CK666 group (0.737±0.031) were significantly decreased compared to those in their respective control groups (all P<0.01). Shape of AM: Long filopodia protruding and plentiful dorsal ruffle can be seen in AM from the healthy control group; AM pseudopods extension and dorsal ruffle reduced in the health CK666 group; there were pseudopods and dorsal ruffle defects in the COPD group and the COPD CK666 group. Positive correlations existed between the proteins of Arp2, F-actin with MFI. Positive correlations also existed between the Manders' overlap coefficient of Arp2 and phalloidin' coefficient with MFI. Conclusion: Decreased activity of Arp2/3 complex leads to low phagocytosis of AM in COPD mice, and AM in COPD mice is more sensitive to Arp2/3 complex inhibitor.

[Effects of TLR4-PI3K-Rac1 pathway on cytoskeleton rearrangement and phagocytosis of macrophage].

Objective: To investigate the effects of Toll-like receptor 4 (TLR4)-phosphatidylinositol 3-kinase (PI3K) -Ras-related C3 botulinum toxin 1 (Rac1) signaling pathway on macrophage cytoskeleton rearrangement and phagocytosis. Methods: Mouse macrophage cell line RAW264.7 was divided into blank group, negative control group and TLR4-RNA interference (RNAi) group. The lentivirus carrying TLR4 short hairpin RNA (shRNA) and nonsense control sequence were respectively transfected into TLR4-RNAi group and negative control group. The cells in blank group were not transfected. The silencing efficiency of TLR4 was detected by Western blot. Real-time fluorescence quantitative PCR was used to detect the expression of PI3K and Rac1 mRNA in each group. The expressions of PI3K, p-Rac1 and Rac1 protein were detected by Western blot. Cytoskeleton was observed by laser scanning confocal microscopy. Mean fluorescence intensity (MFI) and the percentage of cells phagocytosing flurescein inothiocyanate-labeled Eseherichina coli (FITC-E.coli) (Phagocytosed cell %) were detected by flow cytometry. Results: The RAW264.7 cells can be successfully transfected by TLR4-shRNA lentivirus, and the transfection efficiency ranged from 80% to 90%. The silencing efficiency of TLR4 gene was (63±4)%. After silencing the TLR4 gene, the relative expression of TLR4 mRNA and protein (0.20±0.03, 0.37±0.04), PI3K mRNA and protein (0.64±0.06, 0.75±0.06), Rac1 mRNA, protein and p-Rac1 protein (0.75±0.04, 0.76±0.01, 0.74±0.05) in TLR4-RNAi group were significantly lower than those in negative control group and blank group (all P<0.01). The change of cytoskeleton: after silencing the TLR4 gene, the celluar pseudopods were short and stiff, with the impaired capacity of phagocytosing FITC-E.coli. Cells in blank group and negative control group extended good pseudopodia, and the capacity of phagocytosing FITC-E.coli was normal. The MFI and Phagocytosed cells % of TLR4-RNAi group[(7 453±564), (70.20±2.27)%]were significantly lower than those in the blank group and the negative control group (all P<0.01). Positive correlations were existed between mRNA, protein expression of TLR4, PI3K, Rac1 and MFI, Phagocytosed cells% (all P<0.05) in all groups. Conclusion: TLR4-PI3K-Rac1 pathway involves in the cytoskeleton rearrangement and impairs the phagocytosis of macrophages.

[Effects of PI3Kδ-RhoA pathway on phagocytosis defect of alveolar macrophages in a mouse model of chronic obstructive pulmonary disease].

Objective: To investigate effects of Phosphoinositide3-Kinases (PI3Kδ)-Ras homolog gene family member A(RhoA) pathway on phagocytosis deficiency of alveolar macrophages (AMs) in a mouse model of chronic obstructive pulmonary disease (COPD). Methods: Twenty mice were exposed to cigarette smoking to establish the COPD model, with 20 mice as the control group. AMs were isolated from lung tissue by discontinuous density gradient centrifugation and then divided into a healthy control group, a COPD group, a healthy IC87114 group and a COPD IC87114 group. The culture of IC87114 group was mixed with a final concentration of 1 nmol/L IC87114 for 24 hours. Mean fluorescence intensity (MFI) and the positive percent of AMs engulfing flurescein isothiocyanate-labeled Escherichina coli (FITC-E.coli) (AM%) were detected by flow cytometry. Real-Time PCR(RT-PCR)and Western blot were applied to detect mRNA and protein. G-LISA RhoA Kit was used to detect the activity of RhoA, and laser scanning confocal microscopy was used to observe the cytoskeleton structure of AMs. Results: Phagocytosis of AM: MFI and AM %in the COPD group [(4 512±517), (32.2±4.6)%] were decreased than those in the healthy control group [(9 857±1 042), (68.0±4.0)%, all P<0.01]. Compared with the COPD group, MFI and AM% in the COPD IC87114 group [(6 894±472), (50.6±2.1)%] were increased (all P<0.01). The expressions of mRNA and protein of PI3Kδ in the COPD group (3.14±0.54, 0.84±0.08) were increased than those in the healthy control group (1.00±0.00, 0.57±0.07) (all P<0.01). Compared with the COPD group, the expressions of mRNA and protein of PI3Kδ in the COPD IC87114 group (1.52±0.28, 0.66±0.13) were decreased (all P<0.01). The RhoA mRNA, protein and activity in the COPD group (0.70±0.07, 0.41±0.10, 0.70±0.06) were decreased compared to those in the healthy control group (1.00±0.00, 0.56±0.09, 1.19±0.09) (all P<0.01). Compared with the COPD group, the expression of mRNA, protein and activity of RhoA in the COPD IC87114 group(0.91±0.08, 0.48±0.06, 0.86±0.06) were increased (P<0.01, P<0.05). Cytoskeleton of AM: The pseudopods of the healthy control group and the healthy IC87114 group extended well, and the ability of phagocytosing FITC-E.coli was intact, but there were some defects in the COPD group. Compared with the COPD group, the COPD IC87114 group was better, both in phagocytosing and extending of pseudopods. Negative correlations existed between the mRNA, protein of PI3Kδ with mRNA, protein and activity of RhoA. Negative correlations also existed between the mRNA, protein of PI3Kδ with MFI, but positive correlations between RhoA and MFI were observed in all groups. Conclusion: The phagocytosis of AMs in COPD mice was defective, with abnormal rearrangement of the cytoskeleton. PI3Kδ negatively regulated RhoA, while PI3Kδ over activation resulted in decreasing activity of RhoA and then induced abnormal cytoskeleton rearrangement in AMs, which led to phagocytosis deficiency.IC87114 inhibited PI3Kδ activation, improved the activity of RhoA and partly recovered phagocytosis of AMs.

[Mechanisms of cytoskeleton and PI3Kδ-RhoA in fine particulate matter deteriorating phagocytosis defect of alveolar macrophage in mice with chronic obstructive pulmonary disease].

Objective: To explore the mechanism of cytoskeleton and PI3Kδ-RhoA in fine particulate matter deteriorating phagocytosis defect of alveolar macrophage (AM) in chronic obstructive pulmonary disease (COPD) mice. Methods: Forty mice were randomly divided into four groups: health control group, COPD group, health PM2.5 group, COPD PM2.5 group and with ten in each group. A mouse model of COPD was established by cigarette smoke exposure, and health PM2.5 group and COPD PM2.5 group mice were given PM2.5 (588 µg/m(3)) aerosol inhalation for 90 days. AM were isolated from lung tissue by discontinuous density gradient centrifugation. Mean fluorescence intensity (MFI) and the percent of alveolar macrophage engulfing flurescein isothiocyanate-labeled Escherichia coli (FITC-E.coli) AM (AM%) were detected by flow cytometry. The mRNA and protein expression were measured by real time polymerase chain reaction (RT-PCR) and Western blot. The activity of RhoA was measured by GTPase linked immunosorbent assay (G-LISA) Kit. Cytoskeleton was observed by laser scanning confocal microscopy. Results: The MFI and the AM% in COPD group [4 512±517, (32.19±4.57)%] and health PM2.5 group [7 631±585, (50.78±4.58)%] were significantly lower than those in health control group [9 857±1 042, (68.53±2.88)%], while those in COPD PM2.5 group [3 121±393, (21.90±2.58)%] were lower than those in COPD group (all P<0.01). The mRNA and protein of PI3Kδ in COPD group (3.41±0.54, 0.84±0.08)and health PM2.5 group (1.52±0.35, 0.71±0.11) were higher than those in health control group (1.00±0.00, 0.57±0.07) (all P<0.05), and in COPD PM2.5 group (5.53±0.42, 1.17±0.25), the above parameters were remarkably increased as compared to those in COPD group (all P<0.01). The mRNA, protein and activity of RhoA in COPD group (0.70±0.07, 0.41±0.10, 0.70±0.06) and health PM2.5 group (0.84±0.06, 0.46±0.11, 0.87±0.07) were lower than those in health control group (1.00±0.00, 0.56±0.09, 1.19±0.09) (all P<0.05), and above parameters of COPD PM2.5 group (0.42±0.05, 0.31±0.06, 0.44±0.04) were significantly lower than COPD group (all P<0.01). Cytoskeleton of AM: long and dense filopodia and membrane fold could been seen clearly around the AM of health control group; in COPD group and health PM2.5 group, short and sparse filopodia and slightly deformed AM can been seen. Filopodia remarkably decreased and rigid cells with impaired capacity of engulfing FITC-E.coli can be generally observed in COPD PM2.5 group. Negative correlations were existed between PI3Kδ mRNA, protein and RhoA mRNA, protein, activity in all groups (all P<0.01). Negative correlations were existed between PI3Kδ mRNA, protein and MFI, and positive correlations were existed between RhoA mRNA, protein, activity and MFI in all groups (all P<0.05). Conclusion: Fine particulate matter (PM2.5) can deteriorate the phagocytosis of AM from COPD mice through over activating PI3Kδ and inhibiting the activity of RhoA then causing cytoskeleton abnormal rearrangement.

[Effects of mTOR-Cdc42 signaling pathway on phagocytosis of alveolar macrophages in chronic obstructive pulmonary disease mice].

Objective: To investigate effects of mammalian target of rapamycin (mTOR)- cell division cycle 42 (Cdc42) signaling pathway on phagocytosis of alveolar macrophages (AMs) in chronic obstructive pulmonary disease (COPD) mice. Methods: Forty mice were randomly divided into control group and model group. Each group contained 20 mice. COPD model group were established by cigarette smoking exposure. AMs were isolated from lung tissue by discontinuous density gradient centrifugation. AMs from control group were divided into health control group and rapamycin control group while AMs from model group were divided into COPD group and rapamycin COPD group. The AMs from rapamycin control group and rapamycin COPD group were incubated with a final concentration of 10 nmol/L rapamycin for 24 hours. Mean fluorescence intensity (MFI) and the positive percent of alveolar macrophage engulfed flurescein isothiocyanate-labeled Escherichina coli (FITC-E.coli) AM (AM%) were detected by flow cytometry. Real time PCR (RT-PCR) and Western blot were applied to detect mRNA and protein. The activity of Cdc42 was detected by G-LISA Small GTPase Activation Assays (G-LISA) Cdc42 Kit. The cytoskeleton structure of AM was observed by laser scanning confocal microscopy. Results: MFI and AM% in COPD group were decreased than those in health control group[4 060±590 vs 9 190±988 and (28.65±1.26)% vs (67.50±4.56)%]; Compared with COPD group, MFI and AM% in rapamycin COPD group[4 856±762, (38.31±1.71)%]were increased (all P<0.05). The expression of mRNA, protein and activity of mTOR in COPD group were increased than those in health control group[(2.62±0.46, 1.30±0.52, 1.46±0.43) vs (1.00±0.00, 0.48±0.27, 0.58±0.26)]; compared with COPD group, the expression of mRNA, protein and activity of mTOR in rapamycin COPD group (1.40±0.36, 0.90±0.66, 0.92±0.28) were decreased (all P<0.05). The Cdc42 mRNA, protein and activity in COPD group were higher than those in health control group[(2.56±0.50, 1.61±0.37, 0.46±0.09) vs (1.00±0.00, 0.67±0.22, 0.30±0.07)](all P<0.01); compared with COPD group, the expression of mRNA, protein and activity of Cdc42 in rapamycin COPD group (1.38±0.34, 0.91±0.48, 0.36±0.06) were decreased (all P<0.01). Filopodia protruding can not be seen in the cytoskeleton of AMs from health control group and rapamycin control group; some filopodia protruding can be seen in AM from COPD group; some long filopodia protruding can be seen in AM from rapamycin COPD group. Negative correlations were existed between the mRNA, protein and activity of mTOR, Cdc42 and MFI in all group. Conclusions: mTOR-Cdc42 signaling pathway is activated and related to phagocytosis deficiency of AM in COPD. It can be inferred that the pathway is involved in the pathogenesis of COPD.

[Effects and mechanisms of Notch signaling pathway on immune imbalance in chronic obstructive pulmonary disease].

Objective: To explore the role of Notch signaling pathway on immune imbalance in chronic obstructive pulmonary disease (COPD). Methods: Thirty BALB/c mice were randomly assigned into the healthy control group, COPD group, and COPD Gamma secretase inhibitors (GSI) group. Cigarette smoke exposure was used to establish the COPD model. T cells were enriched by filtering through nylon wool columns. The proportion of spleen-derived T-lymphocyte subsets was detected by flow cytometry. Real-time quantitative polymerase chain reaction (RT-PCR) and Western blot (WB) were used to detect the expression of splenic T cells' Notch1 and its downstream Hes1 mRNA and protein respectively. Results: The percentage of Th1, Th17 and Treg cells in CD4＋T cells of COPD mice (13.20±0.95, 10.22±0.45, 0.41±0.09)% were significantly increased compared with the healthy control group (8.07±0.44, 5.98±0.26, 0.26±0.05)%(all P<0.01). The proportion of Th1 and Th17 cells in COPD GSI group mice (9.48 ± 0.66, 7.70 ±0.39)% were significantly reduced compared with COPD group (all P<0.01). Ratio of Th1/Th2 in COPD group (18.70±4.12) was significantly increased compared with the healthy control group (12.63±1.91) (P<0.01), the percentage of Th17 and Treg in CD4＋ T cell increased 71% and 58% respectively. GSI decreased the ratios of Th1/Th2 and Th17/Treg (all P<0.01). Notch1 receptor and its downstream Hes1 mRNA expression (5.15±0.77, 1.92±0.32) and protein expression (0.85±0.04, 0.16±0.02) of COPD mice were significantly increased compared with the healthy group respectively [(1.00 ± 0.00, 1.00 ± 0.00) and (0.17±0.01, 0.09±0.01)] (all P<0.01). GSI significantly inhibited the expression of mRNA and protein in Notch1 and its downstream Hes1 (all P<0.01). Notch1 and Hes1 mRNA and protein expressions were correlated positively with Th1 and Th17 cells and negatively correlated with Th2 and Treg cells in COPD group(all P<0.05). Conclusion: In COPD mice, there was T-lymphocyte subsets imbalance, such as the increased of Th1, Th17 proinflammatory cells, Notch1 and its downstream Hes1 mRNA and protein levels were increased, and was associated with T-lymphocyte subsets imbalance. GSI could partially inhibit Notch1, Hes1 expression and Th1 and Th17 cells, and thus Notch signaling pathway was involved in the immune disorder of COPD mice.

[Effect of PM2.5 on the level of nuclear factor erythroid-2 related factor 2 in chronic obstructive pulmonary disease mice and its relationship with oxidative stress].

OBJECTIVE: To explore the effects of fine particulate matter on the level of nuclear factor erythroid-2 related factor 2 (Nrf2) in pulmonary tissues of chronic obstructive pulmonary disease (COPD) mouse models and its relationship with oxidative stress. METHODS: Totally 40 BALB/c mice were randomly divided into normal control group, normal PM2.5 group, COPD control group and COPD PM2.5 group.COPD mice were established using exposure of cigarette smoking.PM2.5 (20 mg/kg) was intratracheally instilled in PM2.5 group mice.Mice pulmonary function was measured by mice noninvasive body plethysmograph and lung histopathology was observed in normal control group and normal PM2.5 group mice.The mRNA and protein expression of Nrf2 was measured with real-time polymerase chain reaction (PCR) and Western blot methods.Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry.Glutathione peroxidase (GSH-PX) was measured by improved Hafeman colorimetry and malondialdehyde (MDA) by thiobarbiturieacid colorimetry. RESULTS: Nrf2 mRNA and protein in normal control group, normal PM2.5 group, COPD control group and COPD PM2.5 group were 1.00, 4.46±0.42, 4.93±0.63, 6.41±0.35 and 0.92±0.08, 1.23±0.07, 1.20±0.09, 1.43±0.10.Nrf2 mRNA and protein in COPD control group were increased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively higher than each control group.Comparing to normal PM2.5 group, the Nrf2 mRNA and protein in COPD PM2.5 group were increased (all P<0.01). TAC and GSH-PX in each group were (5.1±0.4), (2.9±0.4), (3.3±0.3), (1.8±0.3) and (13.4±0.5), (9.9±0.7), (9.8±0.7), (7.0±0.6) U/mgpro.TAC and GSH-PX in COPD control group were decreased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively lower than each control group.Comparing to normal PM2.5 group, the Nrf2 mRNA and protein in COPD PM2.5 group were decreased (all P<0.01). MDA in each group were (2.9±0.4), (4.8±0.5), (4.5±0.3), and (6.2±0.4) nmol/mgpro.MDA in COPD control group were increased than those in normal control group while those in normal PM2.5 group and COPD PM2.5 group were respectively higher than each control group.Comparing to normal PM2.5 group, the MDA in COPD PM2.5 group were increased (all P<0.01). Positive correlations were observed between Nrf2 mRNA, protein and MDA, while negative correlations were observed between Nrf2 mRNA, protein and TAC, GSH-PX in all groups (all P<0.05). CONCLUSIONS: PM2.5 can induce Nrf2 expression and aggravate oxidative stress in COPD mice.The increased expression of Nrf2 is closely associated with oxidative stress.

[Inhibitory effects of codonopsis pilosula polysaccharides on the deterioration of impaired phagocytosis of alveolar macrophage induced by fine particulate matter in chronic obstructive pulmonary disease mice].

OBJECTIVE: To investigate the inhibitory effects of codonopsis pilosula polysaccharides (CPP) on the deterioration of impaired phagocytosis of alveolar macrophage (AM) induced by fine particulate matter with a mean aerodynamic diameter ≤2.5 µm (PM2.5) in chronic obstructive pulmonary disease (COPD) mice. METHODS: Sixty BALB/c male mice were randomly divided into control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group. COPD mice were established using exposure of cigarette smoking. Meanwhile PM2.5 group, PM2.5 COPD group and CPP+ PM2.5 COPD group were exposed to PM2.5 (770 µg/m(3)) for 90 days. CPP COPD group and CPP+ PM2.5 COPD group were fed with CPP (300 mg/kg) for 90 days whilst other groups were fed with isovolumetric saline. After the models were established, mice peak inspiratory flow (PIF) and peak expiratory flow (PEF) were measured by noninvasive body plethysmograph and lung histopathology and mean linear intercept (MLI) were observed. AMs were isolated from lung tissue by discontinuous density gradient centrifugation. Mean fluorescence intensity (MFI) and the ability of AM phagocytosing flurescein isothiocyanate-labeled Escherichia coli (FITC-E.coli) (AM%) were detected by flow cytometry. Total antioxidative capacity (TAC) was measured by O-phenanthroline colorimetry. Malondialdehyde (MDA) was measured by thiobarbiturieacid colorimetry and glutathione peroxidase (GSH-PX) by improved Hafeman colorimetry. RESULTS: MFI in control group, COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group were 10 267±1 358, 4 817±399, 8 469±240, 3 176±501, 5 886±516 and 4 067±453. AM% in each group were (69.0±5.4)%, (30.7±3.0)%, (51.5±2.4)%, (20.4±3.5)%, (38.7±2.6)% and (28.7±4.3)%. MFI and AM% in COPD group and PM2.5 group were decreased than those in control group while those in PM2.5 COPD group were lower than in COPD group (all P<0.01). Comparing to COPD group and PM2.5 COPD group respectively, MFI and AM% in CPP COPD group and CPP + PM2.5 COPD group were increased (all P<0.01). TAC and GSH-PX in each group were (17.99±0.09), (6.83±0.36), (13.84±1.12), (3.61±0.29), (8.80±0.26), (5.43±0.30) U/mg protein and (84.3±5.7), (46.5±2.6), (62.0±2.2), (32.4±3.8), (53.4±4.0), (42.4±4.0) U/mg. TAC and GSH-PX in COPD group and PM2.5 group were lower than those from control group while those from PM2.5 COPD group were decreased than in COPD group (all P<0.01). Comparing to COPD group and PM2.5 COPD group, TAC and GSH-PX in CPP COPD group and CPP+ PM2.5 COPD group were increased respectively (all P<0.01). MDA in each group were (1.74±0.37), (2.73±0.22), (2.01±0.13), (3.55±0.33), (2.22±0.28) and (2.72±0.44) nmol/mg protein. MDA in COPD group and PM2.5 group were higher than that from control group while that from PM2.5 COPD group was higher than in COPD group; MDA in CPP COPD group and CPP+ PM2.5 COPD group were respectively decreased than those in COPD group and PM2.5 COPD group (all P<0.05). Positive correlations were existed between MFI, AM% and TAC, GSH-PX, while negative correlations were existed between MFI, AM% and MDA in COPD group, PM2.5 group, PM2.5 COPD group, CPP COPD group and CPP+ PM2.5 COPD group. CONCLUSIONS: PM2.5 further impaired the defective phagocytosing capacity of AM and exacerbated oxidative stress in COPD mice. CPP can inhibit these effects. The protection of CPP may be closely related to its antioxidative effects.

The effect of complete radial lateral meniscus posterior root tear on the knee contact mechanics: a finite element analysis.

BACKGROUND: In recent years, with technological advances in arthroscopy and magnetic resonance imaging and improved biomechanical studies of the meniscus, there has been some progress in the diagnosis and treatment of injuries to the roots of the meniscus. However, the biomechanical effect of posterior lateral meniscus root tears on the knee has not yet become clear. The purpose of this study was to determine the effect of a complete radial posterior lateral meniscus root tear on the knee contact mechanics and the function of the posterior meniscofemoral ligament on the knee with tear in the posterior root of lateral meniscus. METHODS: A finite element model of the knee was developed to simulate different cases for intact knee, a complete radial posterior lateral meniscus root tear, a complete radial posterior lateral meniscus root tear with posterior meniscofemoral ligament deficiency, and total meniscectomy of the lateral meniscus. A compressive load of 1000 N was applied in all cases to calculate contact areas, contact pressure, and meniscal displacements. RESULTS: The complete radial posterior lateral meniscus root tear decreased the contact area and increased the contact pressure on the lateral compartment under compressive load. We also found a decreased contact area and increased contact pressure in the medial compartment, but it was not obvious compared to the lateral compartment. The lateral meniscus was radially displaced by compressive load after a complete radial posterior lateral meniscus root tear, and the displacement took place mainly in the body and posterior horn of lateral meniscus. There were further decrease in contact area and increases in contact pressure and raidial displacement of the lateral meniscus in the case of the complete posterior lateral meniscus root tear in combination with posterior meniscofemoral ligament deficiency. CONCLUSIONS: Complete radial posterior lateral meniscus root tear is not functionally equivalent to total meniscectomy. The posterior root torn lateral meniscus continues to provide some load transmission and distribution functions across the joint. The posterior meniscofemoral ligament prevents excessive radial displacement of the posterior root torn lateral meniscus and assists the torn lateral meniscus in transmitting a certain amount of stress in the lateral compartment.