[Research progress on the relationship between air pollution and gestational diabetes].

Gestational diabetes mellitus (GDM) is one of the most common pregnancy complications and has serious implications for the health of mothers and their offspring. In recent years, studies have confirmed that air pollution is one of the main risk factors for diabetes, and there is increasing evidence that air pollution exposure is closely related to the occurrence of gestational diabetes. However, current studies on the association between air pollutant exposure and the incidence of gestational diabetes are inconsistent, and the window period of pollutant exposure is still unclear. Limited mechanistic studies suggest that airborne particulate matter and gaseous pollutants may affect GDM through multiple mechanisms, including inflammation, oxidative stress, disruption of adipokine secretion, and imbalance of intestinal flora. This review summarizes the relationship between air pollutant exposure and the incidence of GDM in recent years, as well as the possible molecular mechanism of the occurrence and development of GDM caused by air pollutants, in order to provide scientific basis for preventing pollutant exposure, reducing the risk of GDM, improving maternal and fetal outcomes and improving the quality of the birth population.

Ultrafast X-Ray Diffraction Visualization of B1-B2 Phase Transition in KCl under Shock Compression.

The classical B1(NaCl)↔B2(CsCl) transitions have been considered as a model for general structural phase transformations, and resolving corresponding phase transition mechanisms under high strain rate shock compression is critical to a fundamental understanding of phase transition dynamics. Here, we use subnanosecond synchrotron x-ray diffraction to visualize the lattice response of single-crystal KCl to planar shock compression. Complete B1-B2 orientation relations are revealed for KCl under shock compression along ⟨100⟩_{B1} and ⟨110⟩_{B1}; the orientation relations and transition mechanisms are anisotropic and can be described with the standard and modified Watanabe-Tokonami-Morimoto model, respectively, both involving interlayer sliding and intralayer ion rearrangement. The current study also establishes a paradigm for investigating solid-solid phase transitions under dynamic extremes with ultrafast synchrotron x-ray diffraction.

Capture Deformation Twinning in Mg during Shock Compression with Ultrafast Synchrotron X-Ray Diffraction.

Deformation twinning plays a vital role in accommodating plastic deformation of hexagonal-close-packed (hcp) metals, but its mechanisms are still unsettled under high strain rate shock compression. Here we investigate deformation twinning in shock-compressed Mg as a typical hcp metal with in situ, ultrafast synchrotron x-ray diffraction. Extension twinning occurs upon shock compression along ⟨112[over ¯]0⟩ and ⟨101[over ¯]0⟩, but only upon release for loading along ⟨0001⟩. Such deformation mechanisms are a result of the polarity of deformation twinning, which depends on directionality and relative magnitude of resolved shear stress and may be common for Mg and its alloys in a wide range of strain rates.

[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.

[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.

[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.

[Expression of platelet miR-223 in coronary artery disease patients and its clinical significance].

Objective: To investigate the correlation of the expression of platelet microRNA-223 (miR-223) with the degree of platelet activation and the severity of coronary lesions in patients with coronary artery disease (CAD). Methods: A total of 204 consecutive patients underwent coronary artery angiography (CAG) at Beijing Anzhen Hospital between April and December 2016 were enrolled. According to CAG results, all the patients were divided into two groups: 112 patients in CAD group and 92 patients in control group. Real-time fluorescent quantitative PCR was used to detect the expression level of platelet miR-223 and mRNA of its target gene P2Y12. Flow cytometry was performed to detect the expression of platelet associated complement-1 (PAC-1). Results: Compared with control group, the expression level of platelet miR-223 in CAD group was higher [(1.27 (1.17, 1.32) vs 0.73 (0.64, 0.79), P<0.001], while the expression level of P2Y12 mRNA was lower [0.75 (0.51, 0.96) vs 1.00 (0.80, 1.64), P<0.001]. The positive expression rate of PAC-1 of CAD group was higher than that of control group (P<0.001). The expression level of platelet miR-223 was negatively correlated with the expression of P2Y12 (r=-0.39, P=0.02), but positively correlated with the expression of PAC-1 (r=0.50, P<0.001). The platelet miR-223 expression level had a positive correlation with the Gensini score in CAD group (r=0.90, P<0.001), but had no significant correlation with platelet aggregation (r=-0.06, P=0.36). Conclusion: The expression level of platelet miR-223 in patients with CAD may be an indicator for monitoring platelet activation and assessing the severity of CAD.

Parametrically Tunable Soliton-Induced Resonant Radiation by Three-Wave Mixing.

We show that a temporal soliton can induce resonant radiation by three-wave mixing nonlinearities. This constitutes a new class of resonant radiation whose spectral positions are parametrically tunable. The experimental verification is done in a periodically poled lithium niobate crystal, where a femtosecond near-IR soliton is excited and resonant radiation waves are observed exactly at the calculated soliton phase-matching wavelengths via the sum- and difference-frequency generation nonlinearities. This extends the supercontinuum bandwidth well into the mid IR to span 550-5000 nm, and the mid-IR edge is parametrically tunable over 1000 nm by changing the three-wave mixing phase-matching condition. The results are important for the bright and broadband supercontinuum generation and for the frequency comb generation in quadratic nonlinear microresonators.

[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 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.

[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.

[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.

[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.

[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.

Bouncing of a dispersive wave in a solitonic cage.

We report the experimental observation of a weak dispersive wave trapped within a cage formed by two solitons in an optical fiber. We show that the dispersive wave bouncing is accompanied by a back-and-forth wavelength conversion of the probe to an idler wave. Additionally, we observed the destruction of the soliton cage when the dispersive wave power is increased, leading to the collision of the solitons.

[Feasibility study on the use of peripheral blood differentially expressed genes for objective classification of chronic subjective tinnitus: a case study on high-frequency tinnitus].

Objective: To explore the feasibility of constructing an objective tinnitus subtype model based on peripheral blood differentially expressed genes (DEGs) using a combination of Weighted Gene Co-expression Network Analysis (WGCNA) and Random Forest algorithm (RF). Methods: From October 2019 to June 2020, peripheral blood DEGs were obtained from 37 patients (from the Third Affiliated Hospital of Sun Yat-sen University)with chronic subjective high-frequency tinnitus (21 unbothersome type, 16 bothersome type) and 20 healthy volunteers through high-throughput sequencing. WGCNA was used to construct gene modules with different expression patterns and analyze their relationships with tinnitus characteristics. Subsequently, RF was employed to build subtype models, which were evaluated by the area under the receiver operating characteristic curve (AUC), accuracy, and F1-score. Results: A total of 12 351 intergroup DEGs were divided into 9 gene modules. Among them, MEblue, MEgreen, and MEbrown showed significant negative correlations with the healthy volunteer group, while MEpink showed a significant positive correlation with the tinnitus distress group. The "Tinnitus vs. Normal" and "Compensatory vs. Decompensatory" subtype models, based on MEblue and MEpink respectively, both had AUCs greater than 0.80, accuracies above 90%, and F1-scores above 0.90, indicating good performance. Conclusions: Peripheral blood DEGs are potential biological indicators for objective classification of subjective tinnitus. The combined application of WGCNA and the Random Forest algorithm should be a viable approach to constructing an objective tinnitus subtype model. However, further exploration and refinement are needed to validate the model's generalizability, cross-dataset performance, and algorithm optimization.

Retrieving quasi-phase-matching structure with discrete layer-peeling method.

An approach to reconstruct a quasi-phase-matching grating by using a discrete layer-peeling algorithm is presented. Experimentally measured output spectra of Solc-type filters, based on uniform and chirped QPM structures, are used in the discrete layer-peeling algorithm. The reconstructed QPM structures are in agreement with the exact structures used in the experiment and the method is verified to be accurate and efficient in quality inspection on quasi-phase-matching grating.

[Etiology and clinical classification of constipation].

The causes of constipation are extremely complex and are still not fully clear. In addition to secondary factors such as organic diseases and drugs, constipation may also be related to genetics, diet, intestinal flora, age, gender and so on. At present, according to the etiology, chronic constipation is divided into primary constipation and secondary constipation. However, there are significant differences among current clinical guidelines in the clinical classification of primary constipation. Some guidelines classify primary constipation as slow-transit constipation (STC), outlet obstruction constipation (OOC), and mixed constipation; however, some guidelines classify primary constipation as STC, defecation disorder (DD), mixed constipation, and normal-transit constipation (NTC); what's more, some even propose types which are different from the above sub-types. There are also differences in the understanding of the relationship between functional constipation (FC) and primary constipation and the classification of irritable bowel syndrome predominant constipation (IBS-C) among various clinical guidelines. By reviewing domestic and international guidelines and relevant literature on constipation, the following conclusions are drawn: primary constipation can be divided into IBS-C and FC, and FC can be further divided into STC, OOC, and mixed constipation; primary constipation should not be confused with FC, nor should IBS-C be classified as FC.

Heterologous expression of the immunomodulatory protein gene from Ganoderma sinense in the basidiomycete Coprinopsis cinerea.

AIMS: FIP-gsi, a fungal immunomodulatory protein found in Ganoderma sinense, has antitumour, anti-allergy and immunomodulatory activities and is regulated by the fip-gsi gene. In this study, we aimed to express the fip-gsi gene from G. sinense in Coprinopsis cinerea to increase yield of FIPs-gsi. METHODS AND RESULTS: A fungal expression vector pBfip-gsi containing the gpd promoter from Agaricus bisporus and the fip-gsi gene from the G. sinense was constructed and transformed into C. cinerea. PCR and Southern blotting analysis verified the successful integration of the exogenous gene fip-gsi into the genome of C. cinerea. RT-PCR and Northern blotting analysis confirmed that the fip-gsi gene was transcribed in C. cinerea. The yield of the FIP-gsi protein reached 314mg kg(-1) fresh mycelia. The molecular weight of the FIP-gsi was 13kDa, and the FIP-gsi was capable of hemagglutinating mouse red blood cells, but no such activity was observed towards human red blood cells in vitro. CONCLUSIONS: The fip-gsi from G. sinense has been successfully translated in C. cinerea, and the yield of bioactive FIP-gsi protein was high. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report using the C. cinerea for the heterologous expression of FIP-gsi protein and it might supply a basis for large-scale production of the protein.

[Acute toxicity of four heavy metal ions to Oncomelania hupensis].

OBJECTIVE: To assess the acute toxicity of Cu2+, Cd2+, Hg2+ and Pb2+ to Oncomelania hupensis. METHODS: Cu2+, Cd2+, Hg2+ and Pb2+ solutions were prepared at five concentrations, and 10 snails were exposed to each concentration for 24, 48, 72 h and 96 h. Then, the inhibition of snail activity and snail death was observed, and the half maximal effective concentration (EC50) and median lethal concentrations (LC50) were estimated. RESULTS: The 24, 48, 72 h and 96 h EC50 values of Cu2+, Cd2+, Hg2+ and Pb2+ were 0.74, 0.56, 0.46, 0.37 mg/L, 4.79, 3.52, 1.70, 1.26 mg/L, 1.90, 1.49, 0.83, 0.76 mg/L and 21.40, 9.98, 7.90, 5.42 mg/L for snails, respectively. The 96 h LC50 values of Cu2+, Cd2+, Hg2+ and Pb2+ were 0.43, 2.96, 1.12 mg/L and 12.22 mg/L for snails, the safe concentrations were 0.004 3, 0.029 6, 0.011 2, 0.122 2 mg/L, respectively. CONCLUSIONS: Cu2+ shows a high acute toxicity to snails, and Cd2+ and Hg2+ exhibit a moderate acute toxicity to snails, while Pb2+ is lowly toxic to snails.