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Objectives: To investigate whether the adenovirus-mediated p53 transfer could enhance the suppression of cervix adenocarcinoma by low-dose (< or =2.0Gy) heavy-ion irradiation., Study Design: HeLa cells were exposed to C-beam, and then infected with AdCMV-p53 or GFP. P53 expression and cell cycle were detected by flow cytometric analysis. Cell apoptosis was observed under a fluorescent microscope with DAPI staining. The survival fraction was determined by colony forming assay., Results: During the observation, the proportion of p53-positive cells in the C-beam with 80 MOI p53 groups was significantly higher than that in control, C-beam only, 80 MOI p53 only, and C-beam with 40 MOI p53 (p<0.05). The proportion of cells in G(1)- or G(2)-phase in the C-beam with p53 groups was significantly higher than that in the C-beam only groups, or p53 only groups (p<0.05). The percentage of apoptotic cells for the C-beam with p53 was significantly higher than that for the C-beam only, or p53 only (p<0.05). The survival fractions for the C-beam with p53 are significantly lower than those for the C-beam only, or p53 only (p<0.05). SF2 for C-beam with 40 or 80 MOI p53 decreased to approximately 10% and approximately 4%, respectively, compared with C-beam alone (approximately 42%)., Conclusions: Adenovirus-mediated p53 transfer could enhance the suppression of cervix adenocarcinoma cells by low-dose C-beam irradiation.

Purpose: To investigate the effect of exogenous wild type p53 (Tp53) on murine melanoma B16 cell apoptosis induced by carbon-ion beam (C-beam) irradiation., Materials and Methods: The murine cell line B16, which has wild-type Tp53 gene status was studied, as well as B16 cells transfected with an adenoviral vector containing the wild-type Tp53 gene (B16/Tp53). Cells were irradiated with C-beam and assayed for cell survival (colony-forming assay), cellular morphology (acridine orange assay), the frequency of apoptotic cells (fluorescence microscopy) and protein expression (Western blot analysis)., Results: The radiosensitivity of B16/Tp53 cells was significantly higher than that of B16 cells. In contrast with Tp53 transfer alone, the combination of C-beam with Tp53 transfer induced a higher proportion of apoptotic cells and micronuclei. After C-beam irradiation, there was no significant increased expression of the cyclin-dependent kinase inhibitor p21 and Tp53 in B16/Tp53 cells compared to B16 cells, but a decreased expression of murine double minute-2 (Mdm2) was observed., Conclusion: The results of this study suggest the potential application of C-beam combined with Tp53 in the treatment of melanoma in human patients.

Objective: The purpose of this study is to investigate the effect of subclinical-dose C-beam irradiation on cell cycle and cell apoptosis in hepatocarcinoma cells., Materials and Methods: The HepG2 cells were exposed to 0-2.0 Gy of either the C beam or a gamma-ray. Cell survival was detected by clonogenic assay. Cell cycle was determined by flow-cytometry analysis. The apoptosis was monitored by fluorescence microscope with DAPI staining. p53 and p21 expression were detected by Western blot., Results: The G0/G1 cells in the irradiated groups were significantly more than those in the control (P<0.05). The C-ion irradiation had a greater effect on the cell cycle of HepG2 cells (including promoting G1-phase and G2-phase arrest) than gamma-ray irradiation. The apoptotic cells induced by C beam were significantly more numerous than those induced by gamma-ray (P<0.05). The carbon ions had a stronger effect on p53 and p21 expression than the gamma-ray irradiation. The survival fractions for cells irradiated by C beam were significantly smaller than those irradiated by gamma-ray (P<0.05)., Conclusion: The subclinical-dose C-beam irradiation significantly suppresses HepG2 cells through cell-cycle arrests and cell apoptosis in contrast to same-dose gamma-ray irradiation.

Objective: To investigate the mechanism of myocardial cell apoptosis during delayed preconditioning induced by Sini decoction (SND)., Method: SD rats were divide into four groups: control, sham, I/R and SND groups. The rats in I/R group, left anterior descending coronary artery (LAD) was occluded for 1h and reperfused for 1 h. The rats in SND group were pretreated with Sini decoction (5 g x kg(-1) x d(-1)) for three days, the last treatment was pretreated 24 h before the index occlusion. Cell apoptosis was measured by flow cytometry, cytochrome C and bcl-xl were detected by Western blotting and the activity of caspase-3 was detected by assay kit., Result: As compared with I/R group, apoptosis rate of myocardial cell, the release of cytochrome C from mitochondria and the activity of caspase-3 were significantly decreased, and the expression of bcl-xl protein was elevated in SND group., Conclusion: The delayed preconditioning induced by Sini decoction decreased myocardial cell apoptosis. The mechanism may be related to the inhibition of mitochondria signal pathway of apoptosis.

Objective: The present study was designed to determine the mechanism of myocardial cell apoptosis during delayed preconditioning induced by Sini decoction (SND)., Methods: Sprage-Dawleytrats were divide into four groups: control, sham, I/R and SND groups. Left anterior descending coronary artery (LAD) of the rats in I/R group were occluded for h and reperfused for h. SND group rates were pretreated with Sini decoction (5 g x kg(-1) x d(-1)) for three days and the last treatment was pretreated 24h before the index occlusion. Cell apoptosis was measured by flow cytometry, mRNA level of bcl-xl/bcl-xs was detected by RT-PCR and apoptosis inducing factor (AIF) was detected by western blotting., Results: SND pretreatment for 24 hours could decrease cell apoptosis, increase bcl-xl-bcl-xs mRNA ratio and decrease release of AIF from mitochondria., Conclusion: The delayed preconditioning induced by SND can decrease myocardial cell apoptosis. The mechanism is related to mitochondria protection.

It has been reported that miR-376a is involved in the formation and progression of several types of cancer. However, the expression and function of miR-376a is still unknown in non-small cell lung carcinomas (NSCLC). In this study, the expression of miR-376a in NSCLC tissues and cell lines were examined by real-time PCR, the effects of miR-376a on cell proliferation, apoptosis and invasion were evaluated in vitro. Luciferase reporter assay was performed to identify the targets of miR-376a. The results showed that miR-376a was significantly downregulated in NSCLC tissues and cell lines. Restoration of miR-376a in NSCLC cell line A549 significantly inhibited cell proliferation, increased cell apoptosis and suppressed cell invasion, compared with control-transfected A549 cells. Luciferase reporter assay showed that c-Myc, an oncogene that regulating cell survival, angiogenesis and metastasis, was a direct target of miR-376a. Over-expression of miR-376a decreased the mRNA and protein levels of c-Myc in A549 cells. In addition, upregulation of c-Myc inhibited miR-376a-induced inhibition of cell proliferation and invasion in A549 cells. Therefore, our results indicate a tumor suppressor role of miR-376a in NSCLC by targeting c-Myc. miR-376a may be a promising therapeutic target for NSCLC. [ABSTRACT FROM AUTHOR]

Objective: To investigate the therapeutic potential of intra-articular hyaluronan-derivative HYADD® 4-G and/or platelet-rich plasma (PRP) in a mouse model of non-invasive joint injury. Methods: Non-invasive axial tibial loading was used to induce joint injury in 10-week-old C57BL/6J mice (n = 86). Mice underwent a single loading of either 6 Newton (N) or 9N axial tibial compression. HYADD® 4-G was injected intra-articularly at 8 mg/mL or 15 mg/mL either before or after loading with or without PRP. Phosphate-buffered-saline was injected as control. Knee joints were harvested at 5 or 56 days post-loading and prepared for micro-computed tomography scanning and subsequently processed for histology. Immunostaining was performed for aggrecan to monitor its distribution, for CD44 to monitor chondrocyte reactive changes and for COMP (cartilage oligomeric matrix protein) as an index for cartilage matrix changes related to loading and cartilage injury. TUNEL assay was performed to identify chondrocyte apoptosis. Results: Loading initiated cartilage proteoglycan loss and chondrocyte apoptosis within 5 days with slowly progressive post-traumatic osteoarthritis (no cartilage degeneration, but increased synovitis and ectopic calcification after 9N loading) at 56 days. Mice treated with repeated HYADD® 4-G (15 mg/mL) or HYADD® 4-G (8 mg/mL) ± PRP or PRP alone exhibited no significant improvement in the short-term (5 days) and long-term (56 days) consequences of joint loading except for a trend for improved bone changes compared to non-loaded joints. Conclusion: While we failed to show an overall effect of intra-articular delivery of hyaluronan-derivative and/or PRP in reversing/protecting the pathological events in cartilage and synovium following joint injury, some bone alterations were relatively less severe with hyaluronan-derivative at higher concentration or in association with PRP. [ABSTRACT FROM AUTHOR]

ABSTRACT In this study, we examined early time-dependent changes in articular cartilage and synovium in response to tibial compression and sought the plausible origin of cells that respond to compression in the healer (LGXSM-6) and non-healer (LGXSM-33) recombinant inbred mouse strains. The right knee of 13-week old male mice was subjected to tibial compression using 9N axial loading. The contralateral left knee served as a control. Knees were harvested at 5, 9, and 14 days post-injury. Histological changes in cartilage and synovium, immunofluorescence pattern of CD44, aggrecan, type-II collagen, cartilage oligomeric matrix protein and the aggrecan neo-epitope NITEGE, and cell apoptosis (by TUNEL) were examined. We used a double nucleoside analog cell-labeling strategy to trace cells responsive to injury. We showed that tibial compression resulted in rupture of anterior cruciate ligament, cartilage matrix loss and chondrocyte apoptosis at the injury site. LGXSM-33 showed higher synovitis and ectopic synovial chondrogenesis than LGXSM-6 with no differences for articular cartilage lesions. With loading, an altered pattern of CD44 and NITEGE was observed: cells in the impacted area underwent apoptosis, cells closely surrounding the injured area expressed CD44, and cells in the intact area expressed NITEGE. Cells responding to injury were found in the synovium, subchondral bone marrow and the Groove of Ranvier. Taken together, we found no strain differences in chondrocytes in the early response to injury. However, the synovial response was greater in LGXSM-33 indicating that, at early time points, there is a genetic difference in synovial cell reaction to injury. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:524-536, 2017. [ABSTRACT FROM AUTHOR]

Objectives Tumour cells produce factors such as interleukin 10 (IL-10), transforming growth factor β1 (TGF -β1) and vascular endothelial growth factor (VEGF) that suppress the function of immune cells or induce apoptosis of immune cells. One of the most important goals of tumour immunotherapy is to antagonize this suppression on immune cells. Ganoderma lucidum polysaccharides ( Gl-PS) may have this potential. The purpose of this study was to determine the antagonistic effects of Gl-PS on the suppression induced by B16F10 melanoma cell culture supernatant (B16F10-CS) on lymphocytes. Methods Gl-PS was used on lymphocytes incubated with B16F10-CS. Enzyme-linked immunosorbent assay was used to determine the levels of IL-10, TGF- β1 and VEGF in B16F10-CS. The MTT assay was used to determine the proliferation of lymphocytes. Immunocytochemistry and Western blot assay were used to determine perforin and granzyme B production in lymphocytes. Key findings There were elevated levels of IL-10, TGF- β1 and VEGF in B16F10-CS. The lymphocyte proliferation, and perforin and granzyme B production in lymphocytes after induction with phytohemagglutinin, as well as lymphocyte proliferation in the mixed lymphocyte reaction, were suppressed by B16F10-CS. This suppression was fully or partially antagonized by Gl-PS. Conclusions B16F10-CS suppressed lymphocyte proliferation and perforin and granzyme B production in lymphocytes after induction with phytohemagglutinin, as well as lymphocyte proliferation in the mixed lymphocyte reaction. This suppression may be associated with elevated levels of immunosuppressive IL-10, TGF- β1 and VEGF in B16F10-CS. Gl-PS had antagonistic effects on the immunosuppression induced by B16F10-CS, suggesting the potential for Gl-PS in cancer immunotherapy. [ABSTRACT FROM AUTHOR]

Abstract: It was demonstrated that oligonucleotides, independent of their base sequence and length, could effectively induce the cellular uptake of mercapto acid-capped CdTe QDs after the oligonucleotides were covalently attached on the surface of the QDs. Following these experimental observations, a conjugate composed of covalently linked anti-survivin antisense oligonucleotides (ASON) and CdTe QDs was designed and synthesized. Then, the survivin mRNA down-regulation and the apoptosis of HeLa cells induced by ASON were studied. Systematic experimental results revealed that CdTe-ASON could effectively induce the apoptosis of HeLa cells, while CdTe QDs offered the possibility to visualize the specific intracellular localization of the CdTe-ASON probes strongly associated with their biological functions. [ABSTRACT FROM AUTHOR]