An A-kinase anchoring protein (ACBD3) coordinates traffic-induced PKA activation at the Golgi.

KDEL receptor (KDELR) is a key protein that recycles escaped endoplasmic reticulum (ER) resident proteins from the Golgi apparatus back to the ER and maintains a dynamic balance between these two organelles in the early secretory pathway. Studies have shown that this retrograde transport pathway is partly regulated by two KDELR-interacting proteins, acyl-CoA-binding domain-containing 3 (ACBD3), and cyclic AMP-dependent protein kinase A (PKA). However, whether Golgi-localized ACBD3, which was first discovered as a PKA-anchoring protein in mitochondria, directly interacts with PKA at the Golgi and coordinates its signaling in Golgi-to-ER traffic has remained unclear. In this study, we showed that the GOLD domain of ACBD3 directly interacts with the regulatory subunit II (RII) of PKA and effectively recruits PKA holoenzyme to the Golgi. Forward trafficking of proteins from the ER triggers activation of PKA by releasing the catalytic subunit from RII. Furthermore, we determined that depletion of ACBD3 reduces the Golgi fraction of RII, resulting in moderate, but constitutive activation of PKA and KDELR retrograde transport, independent of cargo influx from the ER. Taken together, these data demonstrate that ACBD3 coordinates the protein secretory pathway at the Golgi by facilitating KDELR/PKA-containing protein complex formation.

Preclinical development of a novel CCR8/CTLA-4 bispecific antibody for cancer treatment by disrupting CTLA-4 signaling on CD8 T cells and specifically depleting tumor-resident Tregs.

Anti-CTLA-4 antibodies faced challenges due to frequent adverse events and limited efficacy, which spurred the exploration of next-generation CTLA-4 therapeutics to balance regulatory T cells (Tregs) depletion and CD8 T cells activation. CCR8, identified primarily on tumor-infiltrating Tregs, has become a target of interest due to the anti-tumor effects demonstrated by CCR8 antibody-mediated Tregs depletion. Single-cell RNA sequencing analysis reveals that CCR8-positive Tregs constitute a small subset, with concurrent expression of CCR8 and CTLA-4. Consequently, we proposed a novel bispecific antibody targeting CCR8 and CTLA-4 that had the potential to enhance T cell activation while selectively depleting intratumor Tregs. The candidate molecule 2MW4691 was developed in a tetravalent symmetric format, maintaining a strong binding affinity for CCR8 while exhibiting relatively weaker CTLA-4 binding. This selective binding ability allowed 2MW4691 to target and deplete tumor-infiltrating Tregs with higher specificity. In vitro assays verified the antibody's capacity for antibody-dependent cellular cytotoxicity (ADCC) to Tregs with high level of CTLA-4 expression, but not CD8 T cells with relatively low level of CTLA-4 on cell surface. Also, 2MW4691 inhibited the CTLA-4 pathway and enhanced T cell activation. The in vivo therapeutic efficacy of 2MW4691 was further demonstrated using hCCR8 or hCTLA-4 humanized mouse models and hCCR8/hCTLA-4 double knock-in mouse models. In cynomolgus monkeys, 2MW4691 was well-tolerated, exhibited the anticipated pharmacokinetic profile, and had a minimal impact on the peripheral T cell population. The promising preclinical results supported the further evaluation of 2MW4691 as a next-generation Treg-based therapeutics in clinical trials.

Crosstalk between KDEL receptor and EGF receptor mediates cell proliferation and migration via STAT3 signaling.

Hostile microenvironment of cancer cells provoke a stressful condition for endoplasmic reticulum (ER) and stimulate the expression and secretion of ER chaperones, leading to tumorigenic effects. However, the molecular mechanism underlying these effects is largely unknown. In this study, we reveal that the last four residues of ER chaperones, which are recognized by KDEL receptor (KDELR), is required for cell proliferation and migration induced by secreted chaperones. By combining proximity-based mass spectrometry analysis, split venus imaging and membrane yeast two hybrid assay, we present that EGF receptor (EGFR) may be a co-receptor for KDELR on the surface. Prior to ligand addition, KDELR spontaneously oligomerizes and constantly undergoes recycling near the plasma membrane. Upon KDEL ligand binding, the interactions of KDELR with itself and with EGFR increase rapidly, leading to augmented internalization of KDELR and tyrosine phosphorylation in the C-terminus of EGFR. STAT3, which binds the phosphorylated tyrosine motif on EGFR, is subsequently activated by EGFR and mediates cell growth and migration. Taken together, our results suggest that KDELR serves as a bona fide cell surface receptor for secreted ER chaperones and transactivates EGFR-STAT3 signaling pathway.

[L-VDCC autoregulation abnormality contributes to calcium overload in myocardial ischemia-reperfusion injury].

Calcium overload is a vital mechanism of myocardial ischemia-reperfusion injury, which is a hot therapeutic target in cardiovascular research. It has been well recognized that the dysfunction of calcium relevant proteins, including L-type voltage- dependent calcium channel (L-VDCC), sarco/endoplasmic reticulum ATPase 2a (SERCA2a)/phospholamban (PLB), RyR2, Na+/Ca2+ exchanger, Na+/H+ exchanger, etc. contributes to calcium overload in cardiomyocytes during ischemia-reperfusion injury, in which the diastolic calcium concentration is increased and the amplitude of calcium transients is decreased. There are two phases in calcium increase. The early phase is partially mediated by calcium channels, and the latter one is mainly mediated by Na+/Ca2+ exchanger. L-VDCC, a main subtype of calcium channels in myocardium, is involved in calcium overload, but the underlying molecular mechanism is not well elucidated yet. L-VDCC is regulated by intrinsic and extrinsic pathways. PKG and PKA as extrinsic regulators are not proper candidates to increase L-VDCC activity of cardiomyocyte in vitro, whereas the myocardial ischemia-reperfusion injury is highly possible to enhance L-VDCC activity by delaying calcium-dependent inactivation (CDI), advancing calcium-dependent facilitation (CDF), and weakening distal carboxy terminus (DCT) inhibition. Therefore, it is rational to propose that the L-VDCC autoregulation abnormality may play an important role in calcium overload during myocardial ischemia-reperfusion injury.

Research Progress on Laser Powder Bed Fusion Additive Manufacturing of Zinc Alloys.

Zinc, along with magnesium and iron, is considered one of the most promising biodegradable metals. Compared with magnesium and iron, pure Zn exhibits poor mechanical properties, despite its mild biological corrosion behavior and beneficial biocompatibility. Laser powder bed fusion (LPBF), unlike traditional manufacturing techniques, has the capability to rapidly manufacture near-net-shape components. At present, although the combination of LPBF and Zn has made great progress, it is still in its infancy. Element loss and porosity are common processing problems for LPBF Zn, mainly due to evaporation during melting under a high-energy beam. The formation quality and properties of the final material are closely related to the alloy composition, design and processing. This work reviews the state of research and future perspective on LPBF zinc from comprehensive assessments such as powder characteristics, alloy composition, processing, formation quality, microstructure, and properties. The effects of powder characteristics, process parameters and evaporation on formation quality are introduced. The mechanical, corrosion, and biocompatibility properties of LPBF Zn and their test methodologies are introduced. The effects of microstructure on mechanical properties and corrosion properties are analyzed in detail. The practical medical application of Zn is introduced. Finally, current research status is summarized together with suggested directions for advancing knowledge about LPBF Zn.

SUMOylated Golgin45 associates with PML-NB to transcriptionally regulate lipid metabolism genes during heat shock stress.

Golgin tethers are known to mediate vesicular transport in the secretory pathway, whereas it is relatively unknown whether they may mediate cellular stress response within the cell. Here, we describe a cellular stress response during heat shock stress via SUMOylation of a Golgin tether, Golgin45. We found that Golgin45 is a SUMOylated Golgin via SUMO1 under steady state condition. Upon heat shock stress, the Golgin enters the nucleus by interacting with Importin-ß2 and gets further modified by SUMO3. Importantly, SUMOylated Golgin45 appears to interact with PML and SUMO-deficient Golgin45 mutant functions as a dominant negative for PML-NB formation during heat shock stress, suppressing transcription of lipid metabolism genes. These results indicate that Golgin45 may play a role in heat stress response by transcriptional regulation of lipid metabolism genes in SUMOylation-dependent fashion.

Mechanical Properties of Bulk Metallic Glasses Additively Manufactured by Laser Powder Bed Fusion: A Review.

Bulk metallic glasses (BMGs) display excellent strength, high hardness, exceptional wear resistance and corrosion resistance owing to its amorphous structure. However, the manufacturing of large-sized and complex shaped BMG parts faces significant difficulties, which seriously hinders their applications. Laser powder bed fusion (LPBF) is a typical additive manufacturing (AM) technique with a cooling rate of up to 108 K/s, which not only allows for the formation of amorphous structures but also solves the forming problem of complex-shaped BMG parts. In recent years, a large amount of work has been carried out on the LPBF processing of BMGs. This review mainly summarizes the latest progress in the field of LPBF additively manufactured BMGs focusing on their mechanical properties. We first briefly review the BMG alloy systems that have been additively manufactured using LPBF, then the mechanical properties of LPBF-fabricated BMGs including the micro- and nano-hardness, micropillar compressive performance, and macro-compressive and tensile performance are clarified. Next, the relationship between the mechanical properties and microstructure of BMGs produced via LPBF are analyzed. Finally, the measures for improving the mechanical properties of LPBF-fabricated BMGs are discussed. This review can provide readers with an essential comprehension of the structural and mechanical properties of LPBF-manufactured BMGs.

KDEL Receptor Trafficking to the Plasma Membrane Is Regulated by ACBD3 and Rab4A-GTP.

KDEL receptor-1 maintains homeostasis in the early secretory pathway by capturing and retrieving ER chaperones to the ER during heavy secretory activity. Unexpectedly, a fraction of the receptor is also known to reside in the plasma membrane (PM), although it is largely unknown exactly how the KDEL receptor gets exported from the Golgi and travels to the PM. We have previously shown that a Golgi scaffolding protein (ACBD3) facilitates KDEL receptor localization at the Golgi via the regulating cargo wave-induced cAMP/PKA-dependent signaling pathway. Upon endocytosis, surface-expressed KDEL receptor undergoes highly complex itineraries through the Golgi and the endo-lysosomal compartments, where the endocytosed receptor utilizes Rab14A- and Rab11A-positive recycling endosomes and clathrin-decorated tubulovesicular carriers. In this study, we sought to investigate the mechanism through which the KDEL receptor gets exported from the Golgi en route to the PM. We report here that ACBD3 depletion results in greatly increased trafficking of KDEL receptor to the PM via Rab4A-positive tubular carriers emanating from the Golgi. Expression of constitutively activated Rab4A mutant (Q72L) increases the surface expression of KDEL receptor up to 2~3-fold, whereas Rab4A knockdown or the expression of GDP-locked Rab4A mutant (S27N) inhibits KDEL receptor targeting of the PM. Importantly, KDELR trafficking from the Golgi to the PM is independent of PKA- and Src kinase-mediated mechanisms. Taken together, these results reveal that ACBD3 and Rab4A play a key role in regulating KDEL receptor trafficking to the cell surface.

Enhanced Catalytic Effect of Ti2CTx-MXene on Thermal Decomposition Behavior of Ammonium Perchlorate.

Transition metal carbonitrides (MXenes) are promising catalysts due to their special structures. Recently, many studies have shown that MXenes have a catalytic effect on the thermal decomposition of ammonium perchlorate (AP). However, the catalytic effects have not been extensively investigated. Therefore, it is important to illustrate the catalytic mechanisms of pure MXene in AP thermal decomposition. Herein, the catalytic properties of Ti2CTx for ammonium perchlorate (AP) thermal decomposition were investigated by numerous catalytic experiments. The results showed that the high-temperature decomposition (HTD) decreased by 83 °C, and the decomposition heat of AP mixed with Ti2CTx increased by 1897.3 J/g. Moreover, the mass spectrum (MS) data showed that the NH3, H2O, O2, N2O, NO, HCl, and NO2 were formed. In addition, according to the X-ray diffraction (XRD), Raman spectrum, high-resolution transmission electron microscopy (HRTEM), selected area electron diffraction (SAED), and X-ray photoelectron spectra (XPS) results, the Ti2CTx nanosheets can adsorb the gaseous products and react with them in-situ, generating anatase-TiO2 and carbon layers. The Ti2CTx, as-resulted anatase-TiO2, and carbon can synergize and further catalyze the thermal decomposition of AP when both electron and proton transfers are accelerated during AP decomposition.

Structural and electrochemical properties of annealed La1-xYxNi3.55Mno.4Alo.3Coo.75 hydrogen storage alloys.

The effects of the annealing and partial substitution of Y for La on the structural and electrochemical properties of La1-xYxNi3.55Mno.4Alo.3Cro.75 (x = 0, 0.1 and 0.2) alloys were reported in the present work. The single-phased CaCu5-type structure was retained after La was partially substituted by Y. However, the increase of Y content leads to decrease of the lattice parameters and the unit-cell volume. The annealed Lao.9Yo.1Ni3.55Mno.4Alo.3Coo.75 alloys shows typical isometric microstructure, indicating that the composition segregation is improved by annealing. Y substitution for La in the alloys is effective to improve the electrochemical properties at both room temperature and high temperature. A critical substitution content of Y is found at x = 0.1.

Depression of MAD2 inhibits apoptosis and increases proliferation and multidrug resistance in gastric cancer cells by regulating the activation of phosphorylated survivin.

Mitotic arrest-deficient 2 (MAD2) is one of the essential mitotic spindle checkpoint regulators, and it can protect cells from aberrant chromosome segregation. The Mad2 gene is very rarely mutated in many kinds of human cancer, but aberrantly reduced expression of MAD2 has been correlated with defective mitotic checkpoints in several human cancers. We have previously found that the MAD2 expression level is also shown to be associated with the multidrug resistance of tumour cells. In this study, we constructed a small interfering RNA (siRNA) eukaryotic expression vector of MAD2 and downregulated MAD2 expression in the gastric cancer cell line SGC7901 by transfection of MAD2-siRNA. SGC7901 cells stably transfected with the MAD2-siRNA exhibited significantly increased expression of phosphorylated survivin protein and enhanced drug resistance. Furthermore, MAD2-siRNA suppressed the proliferation of SGC7901 cells and inhibited tumour formation in athymic nude mice. This study clearly reveals that downregulation of MAD2 could regulate the cell cycle, increase proliferation, and improve the drug resistance of gastric cancer cells by regulating the activation of phosphorylated survivin. It also suggests both that MAD2 might play an important role in the development of human gastric cancer and that silencing the MAD2 gene may help to deal with the multidrug resistance of gastric cancer cells.

Size-Dependent Structural Properties of a High-Nb TiAl Alloy Powder.

TiAl-based alloys are promising light weight structural materials for high temperature applications in the field of aerospace. Recently, fabrication technologies starting from powders including powder metallurgy and additive manufacturing have been developed to overcome the difficulties in the processing, machining and shaping of TiAl-based alloys. Spherical alloy powders with different particle size distributions are usually used in these fabrication techniques. The purpose of this study is to reveal the size-dependent structural properties of a high-Nb TiAl powder for these fabrication technologies starting from powders. A high-Nb TiAl pre-alloyed powder with nominal composition of Ti-48Al-2Cr-8Nb (at. %) was prepared by the electrode induction melting gas atomization (EIGA) method. The phase structure and morphology of the as-atomized powders were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The size-dependent structural changes of the as-atomized powders with different sizes were studied by differential scanning calorimetry (DSC) and in situ high temperature XRD. It was found that with decreasing the powder size, the content of the γ-TiAl phase decreases and the α2-Ti3Al phase increases. The α2-Ti3Al to γ-TiAl phase transformation was found in the temperature range of 600-770 °C. Based on the present work, the structural characteristics of TiAl powders are strongly dependent on their particle size, which should be considered in optimizing the process parameters of TiAl alloys fabricated from powders.

Structural and Mechanical Characteristics of Cu50Zr43Al7 Bulk Metallic Glass Fabricated by Selective Laser Melting.

In this work, the structural and mechanical characteristics of Cu50Zr43Al7 bulk metallic glass (BMG) fabricated by selective laser melting (SLM) are studied and the impacts from the SLM process are clarified. Cu50Zr43Al7 alloy specimens were manufactured by the SLM method from corresponding gas-atomized amorphous powders. The as-built specimens were examined in terms of phase structure, morphologies, thermal properties and mechanical behavior. The x-ray diffraction and differential scanning calorimetry results showed that structural relaxation and partial crystallization co-exist in the as-fabricated Cu50Zr43Al7 glassy samples. The nano- and micro- hardness and the elastic modulus of the SLM-fabricated Cu50Zr43Al7 BMG were higher than CuZrAl ternary BMGs with similar compositions prepared by conventional mold casting, which can be attributed to the structural relaxation in the former sample. However, the macro compressive strength of the SLM-fabricated Cu50Zr43Al7 BMG was only 1044 MPa mainly due to its porosity. This work suggests that the SLM process induced changes in structural and mechanical properties are significant and cannot be neglected in the fabrication of BMGs.

Celecoxib could reverse the hypoxia-induced Angiopoietin-2 upregulation in gastric cancer.

We investigated the potential effect of Cyclooxygenase-2 (Cox-2) on hypoxia-induced Angiopoietin-2 (Ang-2) expression in gastric cancer cells. Our results revealed that hypoxia augmented Cox-2 and Ang-2 expressions. Also, the hypoxia-induced Ang-2 could be mimicked by CoCl(2) treatment while genestein treatment could partially counteract the hypoxia-induced Ang-2 expression. Celecoxib but not Cox-1 inhibitor sc-560 reversed the hypoxia-induced Ang-2 expression, while this effect could be partially restored by addition of exogenous PGE2. Our findings suggest that the hypoxia-elevated Ang-2 expression in gastric cancer cells may be mediated by both Cox-2-derived PGE2 and HIF-1alpha pathways, while celecoxib could counteract the hypoxia-induced Ang-2 expression.

Mad2beta, an alternative variant of Mad2 reducing mitotic arrest and apoptosis induced by adriamycin in gastric cancer cells.

Mad2beta is an alternative splicing variant of spindle checkpoint gene mad2, which was previously found by us and was related to the drug resistance in gastric cancer cells. In this paper, we explored the molecular mechanisms that Mad2beta variant promoted the formation of multidrug resistance in gastric cancer cells. We found that Mad2beta variant was detected only in the two human drug resistant gastric cancer cell sublines SGC7901/VCR and SGC7901/ADR, and it did not appear in its parental cell line SGC7901 and other detected gastric cancer cell lines. Expressions of Mad2 mRNA and protein in SGC7901 cells transfected with Mad2beta, SGC7901/VCR and SGC7901/ADR were significantly lower than that in SGC7901 cells. Moreover, SGC7901 cells overexpressing Mad2beta variant became more resistant to adriamycin, vincristine and mitomycin by abrogating mitotic arrest and apoptosis. This suggests that expression of Mad2beta variant decreases the relative expression of efficient MAD2, which may help gastric cancer cells to develop the phenotype of multidrug resistance.

Zinc ribbon domain-containing 1 (ZNRD1) mediates multidrug resistance of leukemia cells through regulation of P-glycoprotein and Bcl-2.

Here, we investigated the role of zinc ribbon domain-containing 1 (ZNRD1) in multidrug resistance (MDR) of leukemia cells and the possible underlying mechanisms. ZNRD1 was found overexpressed in the vincristine-induced MDR leukemia cell HL-60/vincristine moreso than its parental cell HL-60. Up-regulation of ZNRD1 expression could confer resistance of both P-glycoprotein (P-gp)-related and P-gp-nonrelated drugs on HL-60 cells and suppress Adriamycin-induced apoptosis accompanied by decreased accumulation and increased releasing amount of Adriamycin. ZNRD1 could significantly up-regulate the expression of P-gp, Bcl-2, and the transcription of the MDR1 gene but not alter the expression of MDR-associated protein, glutathione S-transferase activity, or intracellular glutathione content in leukemia cells. In addition, inhibition of ZNRD1 expression by RNA interference or P-gp inhibitor could partially reverse ZNRD1-mediated MDR. The further study of the biological functions of ZNRD1 may be helpful for understanding the mechanisms of MDR of leukemia and developing possible strategies to treat leukemia.

[A study on the regulatory mechanisms of mitogen-activated protein kinase phosphatase-1 in hypoxia inducible factor-1 trans-activation].

OBJECTIVE: To investigate the regulatory mechanisms of mitogen-activated protein kinase phosphatase-1 (MKP-1) in hypoxia inducible factor (HIF)-1 trans-activation. METHODS: (1) Gastric cancer cells of the line SGC7901 were cultured, then continued to be cultured in hypoxic environment, and was lysed. The supernatant was collected. Western blotting was used to detect the content of total extracellular signal-regulated kinase (ERK) and phosphorylated ERK. (2) Another SGC7901 cells were cultured with PD98059, inhibitor of ERK passway, or SB203580, inhibitor of p38 passway, in the same manner as above-mentioned. Dual luciferase reporter (DLR) was used to detect the luciferase activity so as to measure the HIF-1 trans-activation. (3) siRNA vector U6M2 plasmid against MKP-1 mRNA was constructed. In another experiment SGC7901 cells were cultured and U6M2 and blank vector U6 were transfected into the cells respectively. 24 hours later, the cells were cultured in hypoxic environment with added PD98059 of different concentrations for 12 hours. Dual luciferase reporter (DLR) was used to detect the luciferase activity HIF-1 trans-activation. (4) Another SGC7901 cells were co-transfected with U6M2, pGL-3SV40HRE vector containing promoter SV40, and pRL-TK (internal control vector). Then PD98059 was added, the cells were lysed, and the activity of fluorescein was tested. (5) SGC7901 cells were cultured, transfected with UdM2 or U6 respectively, and 24 hours later cultured under hypoxia with PD98059 of different concentrations for 12 hours. ELISA was used to examine the VEGF protein concentration in the culture fluid. RESULTS: (1) The content of phosphorylated ERK in the SGC7901 cells increased along with the time of hypoxia, peaked at the 12th hour, and then decreased. However, there was no difference in total ERK expression. (2) After 12 hours of hypoxia, different concentrations of PD98059 inhibited the luciferase activity, however, SB203580 of different concentrations had no effect. (3) 24 hours after transfection, the expression of phosphorylated form of ERK in the SGC7901cells transfected with siRNA plasmid against MKP-1 mRNA was higher compared with that in cells transfected with blank vectors after 12 hour of exposure to hypoxia. (4) PD98059 inhibited the luciferase activity either in U6 cells or in U6M2 cells. Notably, when the PD98059 concentration was above 50 micromol/L, there was no difference in HIF-1 activity between the U6 and U6M2 cells. (5) PD98059 of different concentrations all inhibited the VEGF expression either in U6 cells or in U6M2 cells, and when the concentration of PD98059 was over 50 micromol/L there was no difference in VEGF expression level between the U6 cells and the U6M2 cells. CONCLUSION: In SGC7901 cells, the function of MKP-1 is involved in regulation of HIF-1 trans-activation via inactivation of the ERK pathway.

[Inhibition of the in vitro interaction between hypoxia inducible factor 1alpha and p300 by mitogen-activated protein kinase phosphatase 1].

OBJECTIVE: To investigate the effects of mitogen-activated protein kinase phosphatase-1 (MKP-1) on the interaction between hypoxia-inducible factor (HIF)-1alpha and coactivator p300. METHODS: Prokaryotic expression vector PGEX-4T1 was constructed based on DNA recombination technology. GST fusion proteins were purified by glutathione-agarose beads and transfected into Escherichia coli BL-21. Effects of MKP-1 or PD98059 on in vitro interaction between HIF-1alpha and p300 was determined by Pull-down assay coupled with Western bloting. Effects of MKP-1 on the p300 expression level was detected using Western bloting. RESULTS: (1) GST fusion proteins were expressed in BL-21 Escherichia coli and purified proteins were obtained. (2) Less amount of p300 was pulled down by GST fusion protein from SGC7901-MKP-1 cells than from control cells after 12 hours of exposure to hypoxia. (3) 12 hours after hypoxia, Less amount of p300 was pulled down by GST fusion protein from PD98059 treated cells than from DMSO treated cells or SGC7901 cells. (4) There were no significant differences in p300 expression levels between recombinant MKP-1 transfected cells and control cells after 12 hours of exposure to hypoxia. CONCLUSION: MKP-1 inhibits the in vitro interaction between HIF-1alpha and p300.

[Effects of the spider venom on proliferation of human lung adenocarcinoma cell A549].

BACKGROUND AND OBJECTIVE: The spider venom may inspire new drugs to treat cancer. The aim of this study is to investigate the effects and mechanisms of spider venom on lung adenocarcinoma cell A549. METHODS: The proliferation of lung adenocarcinoma A549 cells was detected by MTT. The apoptosis rate was observed with MTT assay and flow cytometer. The activity of catalase was detected by colorimetry. The malondialdehyde (MDA) content was determined by improved thiobarbituric acid fluorometric method. The expression of P38MAPK protein was analyzed with Western blot. RESULTS: Spider venom can remarkably inhibite the proliferation of lung adenocarcinoma A549 cells, increased activity of catalase and MDA content, down-regulated expression of P38MAPK compared with the control group. CONCLUSIONS: The reduced proliferation of lung adenocarcinoma A549 cells by spider venom is may be associated with the increased of activity of catalase and MDA content and decreased expression of P38MAPK.

MAD2 as a key component of mitotic checkpoint: A probable prognostic factor for gastric cancer.

We studied the subcellular localization of MAD2 in normal human tissues and gastric cancers. MAD2 showed nuclear and cytoplasmic localization in normal tissues such as muscle, testis, thyroid gland, cerebrum, trachea, and skin; blood vessels in some organs were also MAD2+. In normal stomach, MAD2 was expressed mainly in cytoplasm but showed nuclear staining in the majority of gastric cancers. MAD2 was significantly overexpressed in gastric cancer compared with matched adjacent tissues (P < .001), and expression was related to differentiation and other clinical parameters of cancer (P < .001). The cancer/adjacent normal tissue (C/N) ratio of MAD2 expression was higher than 2 and more frequently observed in patients with lymph gland metastasis (P < .05) and related to cancer differentiation. Our findings suggest that the steady-state amount of MAD2 inside cells may serve as a molecular switch in mitotic checkpoint control and that the subcellular localizations of this spindle protein undergo a shift during malignant transformation. The change of MAD2 expression may be involved mainly in gastric carcinogenesis and associated with the prognosis of gastric cancer; a C/N of more than 2 may be associated with the worse prognosis for survival in gastric carcinoma.