Iodide Research Articles (Page 1)

Immune checkpoint blockade (ICB) therapy targeting the PD-1/PD-L1 axis results in poor outcomes in prostate cancer (PCa). PD-L1, the most commonly used predictive marker for the efficacy of PD-1/PD-L1-targeted immunotherapy, appears to be rarely or at low levels expressed in primary androgen-responsive PCa tumors, with higher levels in advanced PCa. PD-L1 expression has not yet been studied regarding the androgen receptor (AR) status. We investigated the effect of hormone stimulation by dihydrotestosterone (DHT) and AR inhibition by enzalutamide on PD-L1 expression in LNCaP and LNCaP-AR+ cells, the latter overexpressing AR. Cells were grown for 24 h under hormone-free conditions and then for 24 h in the presence of DHT (10 nM) and/or enzalutamide (10 μM). Cell viability was assessed by Annexin V and propidium iodi de staining. PD-L1 expression was determined semiquantitatively at the mRNA level. ANOVA and independent t-tests were used to compare experimental results between different treatment modalities. DHT treatment induced some degree of apoptosis in AR-overexpressing LNCaP-AR + cells, but not in parental LNCaP cells. We found low basal expression of PD-L1 in both cell lines, with 2.7-fold higher levels in LNCaP-AR+ cells. DHT treatment increased PD-L1 expression by approximately three-fold in LNCaP cells, while in enzalutamide-treated cells, the expression was lower than the basal level. In LNCaP cells treated concomitantly with DHT and enzalutamide, AR inhibition reduced DHT-induced PD-L1, suggesting an androgen-dependent expression of PD-L1. Unlike in LNCaP cells, androgen stimulation did not increase PD-L1 expression in LNCaP-AR+ cells, and enzalutamide did not affect PD-L1 expression either. Our data reveal that PD-L1 is expressed in an AR-dependent manner in PCa cells, and its expression in AR-overexpressing cells is not modulated by receptor inhibition.

Fully Retargeted, Myeloma-Specific Oncolytic Measles Viruses.

Photoinitiated regio- and stereoselective C(sp2)–H perfluoroalkylation and difluoroacetylation of enamides are developed, furnishing biologically and physiologically privileged fluoro-containing enamide scaffolds.

Long noncoding RNAs (lncRNAs) are important regulators involved in a variety of cancer development. However, the role of Linc00210 in non-small cell lung cancer (NSCLC) remains unknown. This study aims to investigate the clinical value of Linc00210 in NSCLC patients and the biological functions of Linc00210 in NSCLC. Gene expression in NSCLC tissues and cell lines was detected using qRT-PCR or Western blot. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays were conducted to evaluate the effect of Linc00210 on NSCLC cell proliferation. Transwell assay and annexin V-Fluorescein 5-isothiocyanate (FITC)/Propidium Iodide (PI) were done to analyze the effect of Linc00210 on cancer cell invasion and apoptosis, respectively. Luciferase reporter assay and RIP assay were performed to determine the target of Linc00210 and miR-16-5p. Besides, these assays were used to determine reciprocally inhibition of each other-controlled NSCLC cell behaviors. In vivo tumorigenesis experiments were applied to exhibit subcutaneous tumor growth. Linc0021 was highly expressed in NSCLC tissues and cell lines. Knockdown of Linc00210 inhibited cancer cell proliferation and invasion, and increased cell apoptosis, and regulated the expression of Cyclin A1, proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin, Bax, and Bcl-2 in NSCLC cells. Further data showed Linc00210 bound to and directly modulated the miR-16-5p levels. Impressively, overexpression of miR-16-5p suppressed NSCLC cell proliferation and invasion, but increased cell apoptosis, and these behaviors could be overturned by overexpression of Linc00210 in vitro and in vivo. Finally, Linc00210 and miR-16-5p cooperatively controlled expression of protein tyrosine kinase 2 (PTK2), a miR-16-5p target. Linc00210/miR-16-5p/PTK2 signaling suggests a promising novel strategy for anti-NSCLC therapy.

Abstract The inverse temperature (T)‐dependent methods yield single crystals of methylammonium lead halide perovskite with strained lattices. In contrast, the antisolvent diffusion method (antisolvent‐vapor crystallization (AVC)) produces unstrained MAPbBr3 crystals with more uniform growth features and lower density of defects. The powder X‐ray diffraction (XRD) measurements confirm the cubic and the tetragonal structure of the MAPbBr3 and the MAPbI3 samples, respectively. The XRD pole figure measurements (MAPbBr3) reveal a roughly parallel dominant (100) lattice plane to the sample surface. An optimal crystal growth combines growth of a seed on an oriented substrate by the AVC method followed by either the AVC or a T‐dependent method. The estimated resistivity ρ and the density of trap state ntrap values for the MAPbI3 samples are 107 Ωcm and 1010 cm−3, respectively. The X‐ray detection test reveals promising electrical properties of the MAPbI3 material. Results from the Hall measurements in van der Pauw geometry for the MAPbBr3 samples agree with those in the literature: the charge‐carrier concentration of 109−1010 cm−3 and the mobility of 7–289 cm2 V−1 s−1, ntrap of 109−1010 cm−3, and ρ of 107–108 Ωcm. These electrical parameters indicate that the MAPbBr3 samples satisfy requirements for radiation sensors.

To investigate the roles and underlying mechanisms of melatonin in oxygen-glucose deprivation/reoxygenation (OGD/R)-insulted SH SY5Y cells. SH SY5Y cells were cultured for OGD/R stimulation. Cell viability and cytotoxicity were measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide, lactate dehydrogenase (LDH), and Hoechst 33258/propidium iodide (PI) staining assays. The mRNA levels of high mobility group box-1 (HMGB1), tumor necrosis factor α (TNF-α), and inducible nitric oxide synthase (iNOS) were analyzed by quantitative Real Time-PCR assays. Nitric oxide (NO) production was assessed by Griess reagent. Reactive oxygen species (ROS) production was detected by fluorescent probe. Malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) were examined by commercial kits. Cell apoptosis was analyzed by flow cytometry and caspase-3 activity. The protein levels were detected by Western blot. Melatonin enhanced the viability and reduced the death and LDH release of OGD/R exposed SH SY5Y cells. Melatonin repressed the HMGB1, TNF-α, and iNOS mRNA expression, NO production, and nuclear factor κB (NF-κB) activation in OGD/R challenged SH SY5Y cells. Melatonin reduced the ROS, MDA, 4-HNE, and 8-OHdG contents but further enhanced the levels of the nuclear factor E2-related factor-2 (Nrf2) and heme oxygenase (HO-1). Melatonin-increased viability and melatonin-decreased LDH release were also mediated by the blockage of NF-κB or reversed by Nrf2 or HO-1 knockdown. Melatonin exerted antiapoptotic effect on OGD/R treated SH SY5Y cells partly by activating Akt signaling. OGD/R challenged SH SY5Y cell autophagy was also repressed by melatonin, as evidenced by the decreased levels of LC-II and beclin-1 and the increased phosphorylation of mammalian target of rapamycin (mTOR), p70 ribosomal protein S6 kinase (p70S6K), and eukaryotic initiation factor 4E binding protein 1 (4E-BP-1). Melatonin protected SH SY5Y cells from OGD/R induced oxidative stress, inflammation, apoptosis, and autophagy by blocking NF-κB signaling and activating Nrf2/HO-1, Akt, and mTOR/p70S6K/4E-BP-1 pathways, thereby indicating that melatonin is a potential and novel therapeutic drug for ischemic stroke.

Objective To investigate the toxicity and optimal concentration of polybrene on lentiviral vector transduction of bone marrow derived dendritic cells (BMDCs). Methods The BMDCs suspensions in mice were prepared and randomly divided into experimental and control groups. The BMDCs in experimental groups were treated with different concentrations of polybrene, and those in control groups were added with phosphate buffer (PBS). Cell counting kit-8 (CCK-8) assay and annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) staining were performed to detect cells viability and apoptosis. Add to BMDCs suspensions were added with lentiviral vector [multiplicity of infection (MOI)=10] and the mixture was divided into experimental and control groups. The mixture in experimental groups was treated with different concentrations of polybrene, and that in control groups was given PBS. The expression of green fluorescent protein (GFP) was observed by fluorescence microscope every day. After the BMDCs were stimulated to mature, the GFP expression was detected using flow cytometry. The t test was used for comparison between two groups. Results The number of living cells in experimental group was significantly less than in control group when polybrene ≥11 mg/L [The absorbance (A) values were 0.881±0.007 vs. 1.031±0.017, t=7.832, P<0.05]. The GFP expression in experimental groups was significantly higher than in control groups. The highest GFP expression was observed when polybrene at 8 mg/L. Conclusion Polybrane shows dose-dependent toxicity on BMDCs, and in the safe range of concentration, polybrane can significantly increase the transfection efficacy of lentiviral vector to BMDCs and the optimal concentration is 8 mg/L. Key words: Polybrene; Lentiviral vector; Bone marrow derived dendritic cells

Objective To explore the effect of botulinum toxin A (BTXA) on proliferation, apoptosis, migration and expression of related proteins of human hypertrophic scar fibroblasts. Methods The hypertrophic scar was surgically removed and the fibroblasts were isolated and cultured. The effect of different concentrations of BTXA on fibroblasts proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. The apoptosis of fibroblasts after treated with BTXA was observed by Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) staining. The effect of different concentrations of BTXA on migration of fibroblasts was observed by Transwell. The effect of BTXA on B-cell lymphoma 2 (bcl-2), cyclin-dependent kinase 4 (CDK4) and phospho-focal adhesion kinase (FAKp-Tyr397) was tested by Western blotting. Results The proliferation of fibroblasts could be inhibited by different concentrations of BTXA (0.46±0.02 vs. 0.59±0.02 vs. 0.83±0.01 at 24 h, t=11.230, 13.290, P<0.01), and the inhibitory rate was significantly increased [ (1.49±0.09)% vs. (3.62±0.52)% vs. (6.36±0.49)%, t=6.947, 16.810, P<0.01]. The Western blot assay showed that the expression of bcl-2, CDK4 and FAKp-Tyr397 was significantly decreased (t=31.750, 86.240, 50.770, P<0.01). Conclusion BTXA can inhibit the proliferation and migration, and induce the apoptosis of fibroblasts. It is expected to be used in the prevention and treatment of hypertrophic scar. Key words: Botulinum toxin A; Hypertrophic scar; Fibroblasts; Proliferation; Apoptosis; Migration

Objective To detect exosome from gastric cancer cell line MKN-28, MKN-45, SGC-7901 on CD8+ T cell. Methods CD8+ T cell was isolated from health person peripheral blood by density gradient centrifugation and immunomagnetic bread, exosome of SGC-7901, MKN-45 and MKN-28 cell lines was obtained by ultracentrifugation. After coculture exosome and CD8+ T cell with different concentration for 48 h, CD8+ T cell was collected, and analyzed cell cycle by flow cytometry, cell apoptosis by Annexin V-propidium iodide (PI) stain, expression of cytokine by enzyme linked immunosorbent assay (ELISA), gene expression by real-time quantitative polymerase chain reaction (Real-time PCR). Results Exosome was isolated successfully from three gastric cancer cell lines and expressed molecular surface markers TSG101 and Alix. CD8+ T cells were isolated in peripheral blood mononuclear cell (PBMC) with a purity of over 98%. The effect of MKN-28 exosome on apoptosis was dose dependent. After stimulation with 100 mg/L MKN-45 exosome for 48 h, CD8+ T cells secreted IL-2, IL-6, IL-10, IFN-γ [(305.44±42.07) ng/L vs. (129.89±11.07) ng/L, P<0.01; (344.39±25.34) ng/L vs. (149.67±15.23) ng/L, P<0.01; (413.31±38.76) ng/L vs. (35.19±24.95) ng/L, P<0.01; (303.78±44.98) ng/L vs. (277.67±40.66) ng/L, P<0.05], and the difference was statistically significant. After SGC-7901 exosome stimulated, CD8+ T cells secreted IL-4, IL-6, IL-10, IFN-γ [(318.61±41.56) ng/L vs. (238.89±35.84) ng/L, P<0.05; (501.06±55.05) ng/L vs. (149.67 ±15.23) ng/L, P<0.01; (486.75±71.20) ng/L vs. (35.19±24.95) ng/L, P<0.01; (621.00±55.14) ng/L vs. (277.67±40.66) ng/L, P<0.01], the difference was statistically significant. After MKN-28 exosome stimulated, CD8+ T cells secreted IL-2, IL-6, IL-10, IFN-γ [ (411.56±48.62) ng/L vs. (129.89±11.07) ng/L, P<0.01; (403.83±52.29) ng/L vs. (149.67±15.23) ng/L, P<0.01; (678.31±78.02) ng/L vs. (35.19±24.95) ng/L, P<0.01; (751.88± 63.16) ng/L vs. (277.67±40.66) ng/L, P<0.01], and the difference was statistically significant. The expression of IL-10 secreted by CD8+ T cells stimulated by 100 mg/L MKN-28 exosome was about 19.53 times compared to the control group. After MKN-45 exosome stimulated, CD8+ T cells expressed forkhead/winged helix transcription factor P3 (Foxp3), interleukin (IL)-10, interferon (IFN)-γ, phosphatase and tensin homologue deleted on chromosome ten (PTEN), signal transducer and activators of transcription 3 (STAT3) (3.46±0.39 vs. 1.00±0.09, P<0.01; 3.29±0.39 vs. 1.01±0.16, P<0.01; 2.24±0.30 vs. 1.00±0.06, P<0.01; 2.14±0.12 vs. 1.00±0.08, P<0.01; 1.24±0.12 vs. 1.00±0.07, P<0.05). After SGC-7901 exosome stimulated, CD8+ T cell expressed Eomes, Foxp3, IL-10, IFN-γ, 2B4, CD160, GATA3, STAT3, PTEN (2.35±0.17 vs. 1.01±0.13, P<0.01; 3.19±0.21 vs. 1.00±0.09, P<0.01; 2.38±0.31 vs. 1.01±0.16, P<0.01; 5.44±0.71 vs. 1.00±0.06, P<0.01; 3.20±0.31 vs. 1.00±0.12, P<0.01; 3.19±0.13 vs. 1.01±0.16, P<0.01; 1.35±0.17 vs. 1.00±0.07, P<0.05; 1.44±0.18 vs. 1.00±0.07, P<0.05; 1.31±0.15 vs. 1.00±0.08, P<0.05) mRNA; after MKN-28 exosome stimulated, CD8+ T cell expressed Eomes, Foxp3, IL-10, IFN-γ, STAT3, PTEN, CD160, 2B4 (3.36±0.18 vs. 1.01±0.13, P<0.01; 4.35±0.30 vs. 1.00±0.09, P<0.01; 5.40±0.63 vs. 1.10±0.16, P<0.01; 6.43±0.84 vs. 1.00±0.06, P<0.01; 1.54±0.28 vs. 1.00±0.07, P<0.05; 0.78±0.11 vs. 1.00±0.08, P<0.01; 1.68±0.42 vs. 1.01±0.16, P<0.05; 1.31±0.22 vs. 1.01±0.12, P<0.05) mRNA expression. The gene expression of CD8+ T cells stimulated by MKN-28 exosome was relatively more changes, such as Foxp3, IL-10 and IFN-γ. Conclusion Gastric cancer cell line exosome stimulated CD8+ T cells to induce cell apoptosis, and change gene expression and cytokine secretion, but the proportion of negative regulatory molecules was much higher. Cancer exosome interacted with CD8+ T cells, which inhibited its immune function, resulting in tumor immune escape, which may be an important factor to promote disease progression. Key words: Gastric cancer; Exosome; CD8+ T cell; Gene expression; Cytokine

Background: Arsenic trioxide (ATO)-containing therapeutic strategies are widely used in the treatment of acute promyelocytic leukemia (APL). Growing evidence has shown that melatonin enhances the radio- or chemo-sensitivity of numerous cancer cells. However, whether melatonin is capable of enhancing the cytotoxic effects of ATO in APL cells remains unknown. Methods: The present study conducted a 24 h melatonin exposure followed by additional 12, 24 or 48 h ATO exposure in the APL cell line NB4 with or without autophagy-related protein 7 (ATG7) silencing by RNA interference. Cell cytotoxicity was evaluated by Cell Counting Kit-8 (CCK-8) and lactate dehydrogenase (LDH) assays. Cell apoptosis was assessed by Annexin-V/propidium iodide assay and western blotting against cleaved caspase 3, Bax and Bcl-2. Autophagy was evaluated by western blotting against LC3. Results: Pre-treatment with a non-cytotoxic dose of melatonin significantly enhanced ATO-mediated reduced cell viability and increased LDH release. Furthermore, melatonin pre-treatment also enhanced ATO-mediated increase in early and late apoptosis, as well as the expression of Bax and cleaved caspase 3, while further decreasing ATO-mediated reduced expression of Bcl-2. Concomitantly, melatonin pre-treatment increased LC3II expression and enhanced the ATO-mediated elevation in LC3II expression. However, autophagy inhibition by ATG7 silencing blocked the enhancing effects of melatonin on ATO-induced apoptosis and cytotoxicity. These findings indicated that melatonin pre-treatment enhances ATO-induced cytotoxicity by modulating ATG7-mediated autophagy. Conclusions: Melatonin could represent a valuable adjuvant to ATO in APL treatment, particularly in patients with ATO-resistant APL.

Objective To investigate the role and mechanism of circular RNA-vimentin (circ-VIM) in the proliferation and apoptosis of colorectal cancer cells. Methods From December 2016 to December 2017, at Department of General Surgery of The First Affiliated Hospital of Zhengzhou University, the clinical data of 100 patients who underwent radical resection of colorectal cancer and were confirmed by pathological examination after operation were collected. The tumor tissues and corresponding paracancerous tissues (negative control) were also collected. The expression of circ-VIM in the colorectal cancer tissues was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The proliferation of HCT-116 and HT29 colorectal cancer cells was detected by cell counting kit-8 assay. The ratio of apoptosis of HCT-116 and HT29 cells was measured by annexin Ⅴ/propidium iodide double staining assay. The mitochondrial membrane potential of HCT-116 and HT29 cells was examined by 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethyl-imidacarbocyanine iodide (JC-1) assay. The expression changes of protein kinase B and mammalian target of rapamycin were tested by Western blotting. The target miRNA of circ-VIM was predicted by miRDB software. T-test and chi-square test were performed for statistical analysis. Results The expression of circ-VIM in colorectal cancer tissues was 2.387±0.536, which was higher than that in corresponding paracancerous tissues (1.110±0.134), and the difference was statistically significant (t=23.096, P<0.01). And the expression levels of circ-VIM were significantly different in patients with different tumor size, TNM stage and lymph node metastasis (all P<0.05). The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM group was 0.737±0.023 and 0.835±0.025, respectively, which were both higher than those in control group (0.449±0.020 and 0.531±0.019), and the differences were statistically significant (t=20.706 and -15.374, both P<0.01). The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 0.236±0.027 and 0.243±0.019, which were lower than those in control group, and the differences were statistically significant (t=24.557 and -23.197, both P<0.01). The ratio of apoptosis of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was (18.00±1.82)% and (20.80±0.61)%, which was higher than those in control group ((6.64±2.01)% and (7.35±1.36)%), and the differences were statistically significant (t=8.826 and 17.454, both P<0.01). The fluorescence intensity ratio of JC-1 aggregate and JC-1 monomer of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 2.21±0.12 and 1.40±0.11, which was lower than those in control group (14.54±1.00 and 9.24±1.18), and the differences were statistically significant (t=-19.558 and -15.685, both P<0.01), which indicated mitochondrial membrane potential decreased. After treated with lenti-circ-VIM-shRNA, the expression of phosphorylated protein kinase B, phosphorylated mammalian target of rapamycin, B-cell lymphoma-2 and mitochondrial cytochrome C at protein level were all down-regulated, however the expression of cytoplasmic cytochrome C, B-cell lymphoma-2 associated X protein and cleaved caspase-3 at protein level were all up-regulated. When the expression of circ-VIM was up-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was down-regulated. When the expression of circ-VIM was down-regulated, the expression of miRNA-147b, miRNA-4447 and miRNA-3656 was up-regulated. Conclusion The expression of circ-VIM in colorectal cancer is abnormally increased, which is involved in the proliferation and apoptosis of colorectal cancer cells. Key words: Colorectal neoplasms; Cell proliferation; Apoptosis; circ-VIM

Опыт применения метоциния йодида у беременных

Objective To investigate the effects of costimulatory molecule B7-H3 on the proliferation and invasion of human non-small cell lung cancer cell line A549. Methods Flow cytometry was used to detect the expression of B7-H3 at protein level on A549 cells. B7-H3-targeting siRNA was transfected into A549 by lentivirus to construct B7-H3-A549 cells, which were identified with Western blot and qPCR. Differences in proliferation between B7-H3-A549 and B7-H3+ A549 cells were analyzed by CCK8 assay. Flow cytometry was performed to detect the changes in apoptosis and cell cycle after AnnexinⅤ-PE/propidium iodide (PI) staining. Transwell assay was used to evaluate the migration and invasion of B7-H3-A549 and B7-H3+ A549 cells. Expression of apoptosis-related proteins was detected by Western blot. Results (1) B7-H3 was highly expressed on A549 cells. A stable B7-H3-A549 cell line and its control cell line B7-H3+ A549 were successfully prepared. (2) A549 cell proliferation was significantly reduced after knocking down B7-H3 expression. (3) The percentage of early apoptotic cells in B7-H3-A549 cell group was higher than that in B7-H3+ A549 cell group, but no significant difference in the percentages of cells undergoing late apoptosis was found between the two groups. B7-H3-A549 cells were arrested at the G0/G1 phase of cell cycle. (4) Compared with B7-H3+ A549 cells, B7-H3-A549 cells showed suppressed migration and invasion. (5) Enhanced expression of Bad and Caspase-3 and decreased expression of Bcl-2, P-AKT and MMP-9 were detected in B7-H3-A549 cells as compared with those in B7-H3+ A549 cells, but no significant difference in the total AKT was observed. Conclusions Knocking down the expression of B7-H3 molecule in A549 cells could inhibit cell proliferation and invasion, induce cell cycle arrest at G0/G1 phase and promote cell apoptosis. Key words: A549 cell; B7-H3; Invasion; Apoptosis

Objective To investigate the effect of basic fibroblast growth factor (bFGF) on autophagy of nerve cells in rats after traumatic brain injury (TBI). Methods A total of 120 healthy adult male SD rats were randomly divided into sham group, TBI+ vehicle group, and TBI+ bFGF group by random number table method, with 40 rats in each group. PinPoint™ Precision Cortical Impactor was used to simulate the pathological damage after TBI. In the sham group, the dura was exposed without impact. In the TBI+ bFGF group, 250 μg/kg of human recombinant bFGF was given in the nasal cavity 1 hour before the modeling, while in the sham group and TBI+ Vehicle group, the same amount of saline was given in the nasal cavity 1 hour before the modeling. The necrotic cells were observed by propidium iodide(PI)staining 6 hours after injury. The effect of bFGF on the nerve function after TBI in rats was evaluated with modified neurological severity score (mNSS) 24 hours after injury. The water content of brain tissue was measured by dry and wet method 48 hours after injury. The ratio of Beclin-1, P62 protein and microtubule-associated protein 1 light 3 (LC3)-II/I protein was detected by western blot. The volume of brain injury was calculated by integral method of brain tissue section. The positive neuron specific nuclear protein (NeuN) cells were observed by immunofluorescence staining. The apoptotic cells were observed by TUNEL. Results Compared with the sham group [(4.0±1.2)%], the percentage of necrotic cells in TBI+ vehicle group [(54.3±10.1)%] and TBI+ bFGF group [(34.5±10.5)%] increased significantly (P<0.05), but the percentage of necrotic cells in TBI+ bFGF group increased less than that in TBI+ vehicle group (P<0.05). Compared with the sham group [(0.3±0.5)points], the mNSS in the TBI+ vehicle group [(5.8±0.8)points] and TBI+ bFGF group [(4.7±1.1)points] were significantly increased (P<0.05), but the mNSS of TBI+ bFGF group was lower than that of TBI+ vehicle group (P<0.05). Compared with the sham group [(76.7±0.7)%], the water content of brain tissue of TBI+ vehicle group [(79.2±0.5)%] and TBI+ bFGF group [(78.4±1.0)%]were significantly increased (P<0.05), but the water content of TBI+ bFGF group was significantly lower than that of TBI+ vehicle group (P<0.05). Compared with the sham group, protein expression of Beclin-1 and LC3-II/I in TBI+ vehicle group and TBI+ bFGF group were significantly improved (P<0.05), P62 protein expression was significantly decreased (P<0.05), the volume of brain tissue injury was significantly increased (P<0.05), the number of positive NeuN cells increased significantly (P<0.05), and the number of apoptotic cells and apoptotic cells were significantly increased (P<0.05). Compared with the TBI+ Vehicle group, the up-regulation of Beclin-1 protein and LC3-II/I protein ratio was obviously inhibited in the TBI+ bFGF group (P<0.05), the down-regulation of P62 protien was significantly suppressed, the volume of brain tissue injury was significantly decreased, and the number of positive NeuN cells and apoptotic cells was significantly reduced (P<0.05). Conclusion The bFGF can significantly inhibit excessive autophagy in rats after TBI, reduce brain edema, reduce cell apoptosis and necrosis, and improve neural function. Key words: Fibroblast growth factor 2; Autophagy; Brain injuries; Brain edema; Apoptosis

Objective To study the mechanisms of the mixed umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) inhibiting proliferation and inducing apoptosis of C6 glioma cells in vitro. Methods Cysteinyl aspartate-specific protease (Caspase)-8, B cell lymphoma/leukemia-2 associated X protein (bax), B cell lymphoma/leukemia-2 (bcl-2) and Caspase-3 protein expression in C6 cells was analyzed by immunohistochemistry and Western blotting, respectively. Caspase-8, bax, bcl-2 and Caspase-3 mRNA levels were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Cell cycle was detected by flow cytometry. Apoptosis of C6 cells was examined by Annexin V-propidium iodide (PI) staining and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) methods, respectively. Results The mixed UCB-MSCs had a tendency to increase the expression of Caspase-8, bax, Caspase-3 mRNA, and decrease the expression of bcl-2 mRNA. The expression of apoptosis protein and the number of apoptosis cells increased with the increase of the ratio of E/T (E: effect cells namely UCB-MSCs; T: target cells namely C6 cells), showing a proportional dependence relationship; E∶T=(5+ 5)∶1. The relative expression of Caspase-8, bax, bcl-2, and Caspase-3 mRNA in C6 cells was 0.78±0.13, 0.28±0.03, 0.70±0.08, and 0.76±0.12, respectively. When E/T=0∶1, 5∶1, (2.5+ 2.5)∶1, 10∶1, (5+ 5)∶1, the number of apoptosis C6 cells was (2.9±0.6), (3.1±0.5), (14.5±1.8), (14.3±2.2), and (22.8±4.5) cells, respectively. Conclusion After phytohemagglutinin (PHA) stimulation, the mixed UCB-MSCs could inhibit proliferation of C6 cells in vitro, which was related to induction of apoptosis of glioma cells. Key words: Glioma; Umbilical cord blood; Mesenchymal stem cells; Apoptosis; Proliferation

Fabrication and Electrocatalytic Performance of Graphene-fullerene Ammonium Iodide Composite Supported Pd Nanocatalyst for Ethanol Oxidation†

Background: The aim of this study was to investigate the correlation of long non-coding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) with the prognosis in breast cancer patients, and its effect on breast cancer cell proliferation and apoptosis. Methods: A total of 178 breast cancer patients were consecutively recruited, then tumor tissue and the paired adjacent tissue were obtained during surgery for lncRNA SNHG1 determination by quantitative polymerase chain reaction (qPCR). LncRNA SNHG1 expression was also measured in breast cancer cell lines and normal breast epithelial cell line. Subsequently, negative control (NC) overexpression plasmids, lncRNA SNHG1 overexpression plasmids, NC short hairpin RNA (shRNA) plasmids and lncRNA SNHG1 shRNA plasmids were transfected into MDA-MB-453 cells as well as MCF7 cells, and cell proliferation and apoptosis were measured afterward. Results: LncRNA SNHG1 expression in tumor tissue was increased compared with paired adjacent tissue, and it correlated with higher T stage and worse overall survival (OS) in breast cancer patients. LncRNA SNHG1 expression was also elevated in breast cancer cell lines compared with normal breast epithelial cell line. Cell Counting Kit-8 (CCK8) assay revealed that lncRNA SNHG1 overexpression promoted while lncRNA SNHG1 shRNA reduced cell proliferation, and Annexin V-fluorescein isothiocyanate/propidium iodide staining (AV/PI) assay illustrated that lncRNA SNHG1 overexpression decreased while lncRNA SNHG1 shRNA increased cell apoptosis rate. In addition, Western Blot assay disclosed that lncRNA SNHG1 overexpression downregulated while lncRNA SNHG1 shRNA upregulated pro-apoptotic marker (C-Caspase3) expression, and lncRNA SNHG1 overexpression increased while lncRNA SNHG1 shRNA decreased anti-apoptotic marker (p-P38) expression. Conclusions: LncRNA SNHG1 is upregulated in tumor tissue and correlates with higher T stage and worse OS, and it promotes cell proliferation but inhibits cell apoptosis in breast cancer.

Objective To assess the effect of fenretinide (4-HPR) on proliferation, apoptosis and migration of B16F10 and A375 melanoma cells, and to evaluate the effect of liposomes and RGD peptide-modified liposomes on its uptake and therapeutic effects. Methods A film-hydration method was used to prepare 4-HPR liposomes (4-HPRL) , which were modified with RGD peptide to prepare RGD-4-HPRL, and the concentration, particle size, electric potential, drug loading capacity and encapsulation efficiency were measured for 4-HPRL and RGD-4-HPRL. In vitro cultured B16F10 and A375 cells were divided into several groups: 4-HPR group, 4-HPRL group and RGD-4-HPRL group treated with Dulbecco′s minimum essential medium (DMEM) containing 4-HPR bulk drug, 4-HPRL and RGD-4-HPRL respectively at the same concentration of 4-HPR, and control group treated with culture solution at the same volume. After different durations of treatment, cell counting kit-8 (CCK8) assay was performed to evaluate cellular proliferative activity, annexin V/propidium iodide staining to detect apoptosis, and scratch wound healing assay to evaluate the effect of drug treatment on cell migration ability. Then, 4-HPR was replaced by coumarin 6 (C6) to prepare C6 liposomes (C6L) and RGD-C6L, and flow cytometry was conducted to evaluate C6 uptake by B16F10 cells. Statistical analysis was carried out with SPSS22.0 software by one-way analysis of variance (ANOVA) for the comparison among several groups and t test for the comparison between two groups. Results The concentration of 4-HPR in the prepared 4-HPRL solution was over 1 300 mg/L. The encapsulation efficiency and drug loading capacity of 4-HPRL were (95.51 ± 1.22) % and (7.27 ± 0.11) % respectively, and those of RGD-4-HPRL were (95.82 ± 0.81) % and (7.14 ± 0.13) % respectively. The particle size distribution of 4-HPRL and RGD-4-HPRL was uniform, and their average particle size was below 100 nm. CCK8 assay showed that 4-HPR could markedly inhibit the proliferative activities of B16F10 and A375 cells. The cell proliferation inhibition rate of 4-HPRL was higher than that of 4-HPR at the same concentration of 4-HPR (P < 0.01) , and the inhibition rate of RGD-4-HPRL was higher than that of 4-HPRL (P < 0.01 or P < 0.05) and 4-HPR (P < 0.01) . As annexin V/propidium iodide apoptosis assay showed, when the concentration of 4-HPR was 10 mg/L, the total apoptosis rates of B16F10 cells in the control group, 4-HPR group, 4-HPRL group and RGD-HPRL group were (4.44 ± 0.35) %, (28.33 ± 0.66) %, (46.43 ± 0.77) % and (51.33 ± 0.37) % respectively. When the concentration of 4-HPR was 20 mg/L, the total apoptosis rates of A375 cells in the above 4 groups were (4.97 ± 0.62) %, (16.68 ± 3.81) %, (32.62 ± 1.24) % and (44.85 ± 4.92) % respectively. The apoptosis rates of B16F10 and A375 cells were significantly higher in the 4-HPRL group than in the 4-HPR group (both P < 0.01) , and higher in the RGD-4-HPRL group than in the 4-HPRL group (both P < 0.01) and 4-HPR group (both P < 0.01) . Scratch wound healing assay showed that 4-HPR could inhibit scratch healing and migration of B16F10 and A375 cells, and the inhibitory effects of 4-HPRL and RGD-4-HPRL were distinctly superior to those of 4-HPR bulk drug. C6 uptake assay revealed that the fluorescence intensity of C6 in B16F10 cells in the control group, C6 group, C6L group and RGD-C6L group were 2.15 ± 0.28, 8.56 ± 0.36, 20.48 ± 0.13 and 22.55 ± 0.07 respectively, and there were significant differences between the 4 groups (F = 67 194.186, P < 0.01) . Additionally, the fluorescence intensity of C6 was significantly higher in the C6L group and RGD-C6L group than in the C6 group (both P < 0.01) , and higher in the RGD-C6L group than in the C6L Group (P < 0.01) . Conclusions 4-HPR can inhibit the proliferation and migration of A375 and B16F10 cells, and induce their apoptosis. Liposomes and RGD-targeted liposomes can markedly enhance the effect of 4-HPR on melanoma cells. Key words: Melanoma; Cell line, tumor; Fenretinide; Liposomes; Integrin alphaVbeta3; Cell proliferation; Apoptosis; Cell migration assays

Objective To explore the relationship between p53 and signal transducer and activators of transcription 3 (STAT3) gene, and detect the effects of p53 and STAT3 on proliferation, migration, invasion and apoptosis of lung cancer cells. Methods Transfected p53 vector, STAT3 vector, si-p53 or si-STAT3 into A549 cells respectively. The expression of p53 and STAT3 were detected by Western blotting assay. Cell counting kit-8 (CCK-8) assay was performed to evaluate cell proliferation; Transwell assay to measure cell migration and invasion; Annexin V/propidium iodide (AV-PI) double staining and flow cytometry were adopted to detect cell apoptosis. Results The STAT3 protein expression was downregulated when overexpressed p53 protein. Meanwhile, STAT3 protein expression was upregulated when p53 protein was knockdown. After knockdown of p53, the proliferation rate of A549 cells was significantly increased (knockdown group: 0.448±0.034 vs. control group: 0.328±0.011) (P<0.05). The migration (ovexpressed group: 73.3±8.82 vs. control group: 133.0±14.5) and invasion (ovexpressed group: 63.3±8.82 vs. control group: 108.0±11.72) abilities was significantly decreased and the the level of apoptosis (ovexpressed group: 11.0±2.082 vs. control group: 4.0±0.577) was increased when downregulated p53 protein (all P<0.05). However, the above-mentioned functions were disappeared when increased p53 protein and STAT3 protein simultaneously. Conclusion p53 negatively regulates the expression of STAT3, thus inhibiting the ability of proliferation, migration and invasion and increased apoptosis. Key words: Lung cancer; p53; Signal transducer and activators of transcription 3; Cells growth; Cells migration; Cells apoptosis

Objective To investigate the influence of overexpression of Golgi phosphorylation protein 3 (GOLPH3) gene on the chemosensitivity of Cisplatin in human colon cancer HT29 cell line. Methods HT29 cells were divided into four groups: control group, small interfering RNA (siRNA) transfection group, experimental group 1 (10 μmol/L Cisplatin), and experimental group 2 [small interfering RNA (siRNA)-GOLPH3+ 10 μmol/L Cisplatin]. The silencing effect of GOLPH3 gene was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. The tumor cell proliferation and formation ability of HT29 cells were measured respectively by methyl thiazol tetrazolium (MTT) assay and tumorsphere formation assay. Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) flow cytometry was used to detect the apoptosis of each group. The protein expression levels of GOLPH3, permeability glycoprotein (P-gp) and β-catenin in HT29 cells were detected by Western blotting. Sixteen nude mice were randomly divided into 4 groups (4 mice in each group): control group, transfection group, experimental group 1 and experimental group 2. The corresponding colon cancer cells were injected subcutaneously into the axillary tissue. The tumor formation in nude mice was observed within 30 days. Results The relative expression level of GOLPH3 mRNA and protein in the transfection group was significantly lower than in the control group (0.162±0.062 vs. 1.002±0.223; 0.099±0.112 vs. 1.003±0.094) (P<0.01). The cell apoptosis rate in the transfection group was significantly higher than in the control group [(15.520±2.921)% vs. (1.843±1.298)%, P<0.01]. Under cisplatin treatment, the A value (0.746±0.085) and the tumorsphere number (212.800±30.380) in the experimental group 1 were significantly higher than those in the experimental group 2 (0.236±0.071 and 30.750±14.500) (P<0.01). Annexin V-FITC/PI flow cytometry showed that the apoptosis rate of the experimental group 1 was significantly lower than that of the experimental group 2 [(23.890±6.363)% vs. (59.400±2.392)%, P<0.01]. The volume of the axillary subcutaneous tumor in the siRNA transfection group, the experimental group 1 and the experimental group 2 was significantly lower than that in the control group [(1.738±0.121), (1.573±0.224), (0.625±0.189) mm3 vs. (2.083±0.110) mm3,P<0.05 for all]. In the experimental group 2, the tumor volume of the nude mice was significantly less than that in the experimental group 1 in the same period after 15 days (P<0.01). Compared to the control group, the expression levels of GOLPH3, β-catenin and P-gp in the experimental group 1 were significantly upregulated (1.000±0.173 vs. 2.734±0.440, 1.000±0.078 vs. 2.472±0.444, and 1.014±0.346 vs. 4.269±0.454) (P<0.01), and the expression levels of these proteins in the experimental group 2 were significantly down-regulated in the experimental group 1 (0.249±0.084 vs. 2.734±0.440, 0.993±0.052 vs. 2.472±0.444, and 1.842±0.383 vs. 4.269±0.454) (P<0.01). Conclusion The overexpression of GOLPH3 can decrease the chemosensitivity of Cisplatin in colon cancer HT29 cells by activating Wnt/β-catenin signal pathway. Key words: Golgi phosphorylation protein 3; Colon cancer cell; Wnt/β-catenin signal pathway; Chemosensitivity; Cisplatin