Efficacy and safety of immune checkpoint inhibitors with or without radiotherapy in metastatic non-small cell lung cancer: A systematic review and meta-analysis.

Background and purpose: Although immune checkpoint inhibitors (ICIs) have become the first-line treatment for metastatic non-small cell lung cancer (mNSCLC), their efficacy is limited. Meanwhile, recent reports suggest that radiotherapy (RT) can activate the systemic antitumor immune response by increasing the release of antigens from tumor tissues. Therefore, in patients with mNSCLC treated with ICIs, investigations were performed to determine whether the addition of RT improved the outcomes. Furthermore, the adverse events rate was evaluated. Methods and materials: Pubmed, Embase, and Cochrane Library were searched using the keywords "radiotherapy," "immune checkpoint inhibitors," and "non-small cell lung cancer" from the date of inception to 2 May 2022. Randomized controlled trials (RCTs) and nonRCTs (NRCTs) comparing the efficacy and safety of RT combined with ICIs versus ICIs alone in metastatic NSCLC were assessed. The primary outcomes were progression-free survival (PFS) and overall survival (OS), and the secondary outcomes were abscopal response rate (ARR), abscopal control rate (ACR), adverse events rate, and pneumonia rate. The analyses were conducted using the Mantel-Haenszel fixed-effects or random-effects model. The I2 statistic was used to determine heterogeneity, whereas funnel plots and Egger's test were used to assess publication bias. Results: In 15 clinical studies, 713 patients received RT combined with ICIs and 1,275 patients received only ICIs. With regard to PFS and OS, the hazard ratios of RT combined with ICIs were 0.79 (0.70, 0.89) and 0.72 (0.63, 0.82), respectively. In terms of ARR and ACR, the odds ratios (ORs) of RT combined with ICIs were 1.94 (1.19, 3.17) and 1.79 (1.08, 2.97), respectively. Subgroup analyses based on study type (RCT/NRCT), RT target (intracranial/extracranial), number of RT sites (single site), previous ICI resistance (yes/no), and sequencing of RT and ICIs (concurrent/post-RT ICIs) revealed that the addition of RT significantly prolonged PFS and OS. However, subgroup analyses based on radiation dose/fractionation indicated that the addition of hypofractionated RT significantly prolonged OS but not PFS. When grouped according to the level of PD-L1 expression, the addition of RT prolonged PFS only in patients who were PD-L1-negative. Furthermore, subgroup analyses of ARR and ACR signified that the combination therapy resulted in better local control of lesions outside the irradiation field in the hypofractionated RT, extracranial RT, and ICI-naïve subgroups. In terms of adverse events, the addition of RT did not significantly increase the adverse events rate but was associated with a higher pneumonia rate [OR values were 1.24 (0.92, 1.67) and 1.76 (1.12, 2.77), respectively]. Conclusion: Meta-analysis of existing data suggests that the addition of RT can significantly prolong PFS and OS in patients with metastatic NSCLC receiving ICIs. In addition to lesions in the irradiation field, RT can improve the local control rate of lesions outside the irradiation field via immune activation. Combination therapy does not increase the overall risk of adverse reactions, except for pneumonia.

Mechanism, Prevention, and Treatment of Radiation-Induced Salivary Gland Injury Related to Oxidative Stress.

Radiation therapy is a common treatment for head and neck cancers. However, because of the presence of nerve structures (brain stem, spinal cord, and brachial plexus), salivary glands (SGs), mucous membranes, and swallowing muscles in the head and neck regions, radiotherapy inevitably causes damage to these normal tissues. Among them, SG injury is a serious adverse event, and its clinical manifestations include changes in taste, difficulty chewing and swallowing, oral infections, and dental caries. These clinical symptoms seriously reduce a patient's quality of life. Therefore, it is important to clarify the mechanism of SG injury caused by radiotherapy. Although the mechanism of radiation-induced SG injury has not yet been determined, recent studies have shown that the mechanisms of calcium signaling, microvascular injury, cellular senescence, and apoptosis are closely related to oxidative stress. In this article, we review the mechanism by which radiotherapy causes oxidative stress and damages the SGs. In addition, we discuss effective methods to prevent and treat radiation-induced SG damage.

Effect of Dapagliflozin on Intestinal Flora in MafA-deficient Mice.

OBJECTIVE: To investigate the effect of dapagliflozin on intestinal microflora in MafA-deficient mice using an animal model of diabetes. METHODS: Male MafA-deficient mice were administered dapagliflozin (1.0 mg/kg/d) intragastrically for 6 weeks. Mouse body weights and fasting blood glucose levels were measured, and intestinal short-chain fatty acids were measured by gas chromatography. A series of methods was used to analyse the number of primary harmful bacteria in the faeces, and high-throughput sequencing was used to sequence the changes in intestinal flora. RESULTS: The weight of the mice decreased after dapagliflozin gavage, and fasting blood glucose was significantly lower than that in the control group (P < 0.001). Acetic acid and butyric acid contents in the intestinal tracts of the mice increased, and the growth of harmful microorganisms, such as Clostridium perfringens, enterococci, Enterobacteriaceae, and intestinal enterococci, was inhibited. Blautia is a species found in the experimental group and was significantly different from the control and blank groups as determined by the LDA score from highthroughput sequencing. CONCLUSION: Dapagliflozin can reduce fasting blood glucose, decrease body weight, increase short-chain fatty acid content, regulate the intestinal microecological balance of the body and promote blood glucose and energy homeostasis.

Mesenchymal stem cell-based therapy for radiation-induced lung injury.

Since radiotherapy is widely used in managing thoracic tumors, physicians have begun to realize that radiation-induced lung injury (RILI) seriously limits the effects of radiotherapy. Unfortunately, there are still no effective methods for controlling RILI. Over the last few decades numerous studies have reported the beneficial effects of mesenchymal stem cells (MSCs) on tissue repair and regeneration. MSCs can not only differentiate into lung alveolar epithelial cells and secrete anti-inflammatory factors, but they also deliver some vehicles for gene therapy in repairing the injured lung, which provides new ideas for managing RILI. Thus, many scientists have attempted to manage RILI using MSC-based therapy. However, as a novel therapy MSCs still face various limitations. Herein, we shed light on the current understanding of MSC-based therapy for RILI, including the feasibility, molecular mechanisms, animal studies, and clinical research of MSC-based therapy for RILI. We also present an overview of RILI and MSCs.

Overexpression of truncated AIF regulated by Egr1 promoter radiation-induced apoptosis on MCF-7 cells.

It has been demonstrated that gene-radiotherapy can improve the radiotherapy by selectively increasing cells' response to ionizing radiation. Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein, and its C-terminal domain is responsible for the proapoptotic activity. In the present study, we overexpressed truncated AIF on MCF-7 cells by transfection of pcDNA3.1-tAIF (pc-tAIF) and pcDNA3.1-Egr1-tAIF (pc-Egr1-tAIF) plasmids. After MCF-7-tAIF cells were exposed to X-rays, the AIF and tAIF expressions, cell proliferation, apoptosis, cell cycle invasion, cytochrome c (Cyt c) release and activation of caspase-9 were measured by using Western blot, MTT assay, flow cytometry and Matrigel transwell assay, respectively. Our results showed that tAIF expression increased on time- and dose-dependent manners. Both tAIF and radiation can synergistically enhance the apoptosis, cell proliferation inhibition, cell cycle arrest and cell-invasive inhibition. In addition, tAIF overexpression and irradiation increased Cyt c release. However, only irradiation increased caspase-9 activation. Our studies indicated that tAIF overexpression might enhance apoptosis induced by radiation in MCF-7 cells.

Hypoxia- and radiation-induced overexpression of Smac by an adenoviral vector and its effects on cell cycle and apoptosis in MDA-MB-231 human breast cancer cells.

A conditionally replicative adenoviral (CRAd) vector, designated as CRAd.pEgr-1-Smac, that promotes the overexpression of second mitochondria-derived activator of caspase (Smac) when stimulated by hypoxia and radiation was constructed. MDA-MB-231 cells were transfected with CRAd.pEgr-1-Smac and treated with 4-Gy X-rays. The hypoxic status in cancer cells was mimicked with the chemical reagent CoCl2. Smac protein expression was measured by a western blotting assay and cell proliferation was detected with the MTT assay. The cell cycle progression and apoptotic percentage were measured by flow cytometry with PI and Annexin V-FITC staining kits, respectively, following the irradiation of the transfected cells with 4-Gy X-rays. The results showed that CRAd.pEgr-1-Smac was able to increase the Smac protein expression induced by hypoxia and radiation, inhibit cell proliferation and promote apoptosis. Therefore, this method of gene-radiotherapy is indicated to be an ideal strategy for the treatment of breast cancer.

Involvement of endoplasmic reticulum stress in apoptosis of testicular cells induced by low-dose radiation.

The study examined the role of endoplasmic reticulum stress (ERS) and signaling pathways of inositol-requiring enzyme-1 (IRE1), RNA-activated protein kinase-like ER kinase (PERK) and activating transcription factor-6 (ATF6) in apoptosis of mouse testicular cells treated with low-dose radiation (LDR). In the dose-dependent experiment, the mice were treated with whole-body X-ray irradiation at different doses (25, 50, 75, 100 or 200 mGy) and sacrificed 12 h later. In the time-dependent experiment, the mice were exposed to 75 mGy X-ray irradiation and killed at different time points (3, 6, 12, 18 or 24 h). Testicular cells were harvested for experiments. H(2)O(2) and NO concentrations, and Ca(2+)-ATPase activity were detected by biochemical assays, the calcium ion concentration ([Ca(2+)]i) by flow cytometry using fluo-3 probe, and GRP78 mRNA and protein expressions by quantitative real-time RT-PCR (qRT-PCR) and Western blotting, respectively. The mRNA expressions of S-XBP1, JNK, caspase-12 and CHOP were measured by qRT-PCR, and the protein expressions of IRE1α, S-XBP1, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP by Western blotting. The results showed that the concentrations of H2O2 and NO, the mRNA expressions of GRP78, S-XBP1, JNK, caspase-12 and CHOP, and the protein expressions of GRP78, S-XBP1, IRE1α, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP were significantly increased in a time- and dose-dependent manner after LDR. But the [Ca(2+)]i and Ca(2+)-ATPase activities were significantly decreased in a time- and dose-dependent manner. It was concluded that the ERS, regulated by IRE1, PERK and ATF6 pathways, is involved in the apoptosis of testicular cells in LDR mice, which is associated with ERS-apoptotic signaling molecules of JNK, caspase-12 and CHOP.

[Low-dose radiation induces endoplasmic reticulum stress and activates PERK-CHOP signaling pathway in mouse testicular cells].

OBJECTIVE: To explore the correlation of low-dose radiation with endoplasmic reticulum stress and the activation of the PERK-CHOP signaling pathway in mouse testicular cells. METHODS: Healthy Kunming mice were randomly assigned to time-effect (0, 3, 6, 12 and 24 h of irradiation at 75 mGy) and dose-effect (12 h of irradiation at 0, 50, 75, 100 and 200 mGy) groups. The contents of H202 and MDA were measured by colorimetry with the agent kits, the expressions of GRP78, PERK and CHOP mRNA detected by quantitative RT-PCR, and the levels of GRP7B, PERK, phosphorylated PERK (pho-PERK) and CHOP proteins determined by Western blotting and image analysis. RESULTS: After whole-body irradiation of the mice with 75 mGy, the content of H2 02 in the testis tissue was increased with time prolongation, while that of MDA decreased slightly at 3 and 6 h and then increased with the lengthening of time, both increased significantly at 12 and 24 h as compared with those at 0 h (P < 0. 05, P < 0. 01). Apart from reduced levels of GRP78 mRNA at 3 and 24 h and GRP78 protein at 6 h after irradiation, significant increases were found in the mRNA expressions of GRP78 at 12 h, PERK at 3,6, 12 and 24 hand CHOP at 12 and 24 h (P < 0.05, P < 0.01), as well as in the protein levels of GRP78 at 12 and 24 h, pho-PERK at 3, 12 and 24 h and CHOP at 3, 6, 12 and 24 h in comparison with those at 0 h (P < 0. 05, P < 0. 01). No obvious regularity was observed in the change of the PERK protein expression. After 12 h of whole-body irradiation, the content of H202 was increased at 50, 75 and 100 mGy, but decreased slightly at 200 mGy, while that of MDA was increased with dose increasing, with significant increases in the content of H2 02 at 75 and 100 mCy and in that of MDA at 75, 100 and 200 mGy as compared with the 0 mGy group. Apart from the reduced levels of GRP78 mRNA at 50 and 200 mCy, significant increases were found in the mRNA expressions of PERK at 75, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 (P c 0. 05, P < 0.01) as well as in the protein levels of GRP78 at 100 and 200 mGy, pho-PERK at 50, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 mCy as compared with those at 0 mGy (P < 0. 05, P < 0. 01). There were differences in the changes of different protein expressions, but no obvious regularity was seen in the change of the PERK protein expression. CONCLUSION: Low-dose radiation can induce endoplasmic reticulum stress in mouse testicular cells, and activate the PERK-CHOP signaling pathway.

Enhanced effects of TRAIL-endostatin-based double-gene-radiotherapy on suppressing growth, promoting apoptosis and inducing cell cycle arrest in vascular endothelial cells.

This study examined the effects of TRAIL-endostatin-based gene-radiotherapy on cellular growth, apoptosis and cell cycle progression in human vascular endothelial cells ECV304 in vitro. The expression of TRAIL and endostatin protein in ECV304 cells was detected by ELISA after the transfection of recombinant plasmid pshuttle-Egr1-shTRAIL-shES and X-ray irradiation. Then MTT assay was used for determining the cellular proliferation, and flow cytometry (FCM) plus Annexin V and propidium iodide (PI) double-staining or PI single-staining were employed for the detection of apoptosis and cell cycle progression. The results showed that expression of TRAIL and endostatin protein exhibited a time- and dose-dependent change in ECV304 cells after pshuttle-Egr1-shTRAIL-shES transfection in conjunction with irradiation. In the TRAIL-endostatin-based single- or double-gene-radiotherapy, the cell viability declined in a time- and dose-dependent manner, the percentage of cells at G(2)/M phase and apoptotic rate was increased, and the percentage of cells at G(0)/G(1) phase was lowered as compared with those receiving radiotherapy alone. Moreover, TRAIL-endostatin-based double-gene-radiotherapy demonstrated better effects on growth inhibition, promotion of apoptosis and induction of cell cycle arrest in ECV304 cells than single-gene-radiotherapy.

Anti-tumor effect of 125I-UdR in combination with Egr-1 promoter-based IFNγ gene therapy in vivo.

Although (125)I-UdR treatment of malignant tumors in animal models and patients has achieved a certain effect, the short half-life of (125)I-UdR in vivo and its cellular uptake only in S phase of the cell cycle are limiting factors with regard to tumor eradication, and therefore its combination with other applications is a promising strategy in cancer therapy. In this study, we show that (125)I-UdR radionuclide therapy in combination with Egr-1 promoter-based IFNγ gene therapy is more effective than (125)I-UdR therapy alone in suppressing tumor growth and extending survival duration in mice bearing H22 hepatomas. Combined therapy could significantly inhibit cell proliferation and tumor angiogenesis, induce apoptosis and enhance cytotoxic activities of splenic CTL of the mice. Our results suggest that (125)I-UdR in combination with Egr-1 promoter-based IFNγ gene therapy may provide novel approaches for cancer treatment.

Inhibition of human and rat 3beta-hydroxysteroid dehydrogenase and 17beta-hydroxysteroid dehydrogenase 3 activities by perfluoroalkylated substances.

Perfluoroalkylated substances (PFASs) including perfluorooctane acid (PFOA) and perfluorooctane sulfonate (PFOS) have been classified as persistent organic pollutants and are known to cause reduced testosterone production in human males. The objective of the present study was to compare the potencies of five different PFASs including PFOA, PFOS, potassium perfluorooctane sulfonate (PFOSK), potassium perfluorohexane sulfonate (PFHxSK) and potassium perfluorobutane sulfonate (PFBSK) in the inhibition of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 17beta-hydroxysteroid dehydrogenase 3 (17beta-HSD3) activities in the human and rat testes. Human and rat microsomal enzymes were exposed to various PFASs. PFOS and PFOSK inhibited rat 3beta-HSD activity with IC(50) of 1.35 + or -0.05 and 1.77 + or - 0.04 microM, respectively, whereas PFHxSK and PFBSK had no effect at concentrations up to 250 microM. All chemicals tested weakly inhibited human 3beta-HSD activity with IC(50)s over 250 microM. On the other hand, PFOS, PFOSK and PFOA inhibited human 17beta-HSD3 activity with IC(50)s of 6.02 + or - 1.02, 4.39 + or - 0.46 and 127.60 + or - 28.52 microM, respectively. The potencies for inhibition of 17beta-HSD3 activity were determined to be PFOSK>PFOS>PFOA>PFHxSK=PFBSK for human 17beta-HSD3 activity. There appears to be a species-dependent sensitivity to PFAS-mediated inhibition of enzyme activity because the IC(50)s of PFOS(K) for inhibition of rat 17beta-HSD3 activity was greater than 250 microM. In conclusion, the present study shows that PFOS and PFOSK are potent inhibitors of rat 3beta-HSD and human 17beta-HSD3 activity, and implies that inhibition of steroidogenic enzyme activity may be a contributing factor to the effects that PFASs exert on androgen secretion in the testis.

Repetitive exposures to low-dose X-rays attenuate testicular apoptotic cell death in streptozotocin-induced diabetes rats.

To define whether repetitive exposures to low-dose radiation (LDR) can attenuate diabetes-induced testicular cell death, Type 1 diabetic rats were produced by single injection of streptozotocin (STZ). Once hyperglycemia was diagnosed, diabetic rats were treated with and without LDR (25 and 50 mGy X-rays) daily for 4 weeks. Eight and 12 weeks after diabetes onset, testicular apoptotic cell death was examined by flow cytometry with Annexin V/PI staining, Western blotting assay for caspase-3 cleavage, and TUNEL staining for localization of apoptotic cells. Diabetes induced a significant increase in testicular apoptotic cell death, which was able to be attenuated by repetitive exposures to LDR. Diabetes-induced testicular cell death was associated with increased mitochondrial dysfunction, shown by the decreased mitochondrial potential and increased expressions of Bax mRNA and protein. All these changes were significantly attenuated in certain extends by repetitive exposures to LDR. To investigate the mechanisms by which LDR attenuates diabetes-induced testicular apoptotic cell death, serum sex hormone (testosterone, luteinizing hormone and follicle stimulating hormone) levels, and both serum and testicular oxidative damage (lipid peroxides) and antioxidant contents (superoxide dismutase, catalase and glutathione) were measured. Serum sex hormones were significantly decreased in diabetic rats, but not significantly in diabetic rats with multiple exposures to LDR; serum and testicular oxidative damage was significantly increased along with significant decreases in serum and testicular antioxidants in diabetic rats; however, these changes were significantly prevented by repetitive exposures to LDR. Furthermore, diabetic effects on the testicular oxidative damage and cell death were all attenuated by antioxidant N-acetylcysteine. These results suggest that diabetes-induced testicular cell death is probably mediated by increased oxidative stress. LDR protection from diabetes-induced testicular cell death is most likely mediated by its preserving antioxidants.

Increased levels of p53 and PARP-1 in EL-4 cells probably related with the immune adaptive response induced by low dose ionizing radiation in vitro.

OBJECTIVE: This paper is to explore the DNA repair mechanism of immune adaptive response (AR) induced by low dose radiation (LDR), the changes of mRNA levels and protein expressions of p53, ATM, DNA-PK catalytic subunit (DNA-PKcs) and PARP-1 genes in the LDR-induced AR in EL-4 cells. METHODS: The apoptosis and cell cycle progression of EL-4 cells were detected by flow cytometry in 12 h after the cells received the pre-exposure of 0.075 Gy X-rays (inductive dose, D1) and the succeeding high dose irradiation (challenge dose, D2; 1.0, 1.5, and 2.0 Gy X-rays, respectively) with or without wortmannin (inhibitor of ATM and DNA-PK) and 3-aminobenzamid (inhibitor of PARP-1). And the protein expressions and mRNA levels related to these genes were detected with flow cytometry and reverse transcription-polymerase chain reaction in 12 h after irradiation with D2. RESULTS: The mRNA and protein expressions of p53 and PARP-1 in EL-4 cells in the D1 + D2 groups were much lower than those in the D2 groups, and those of PARP-1 in the 3-AB + D2 and the 3-AB + D1 + D2 groups were much lower than those in the D2 and the D1 + D2 groups. The percentage of apoptotic EL-4 cells in the 3-AB + D1 + D2 groups was much higher than that in the D1 + D2 groups, that in the G0/G1 and the G2 + M phases was much higher, and that in the S phase were much lower. Although the ATM and DNA-PKcs mRNA and protein expressions in wortmannin + D1 + D2 groups were much lower than those in the D1 + D2 groups, there were no significant changes in the apoptosis and cell cycle progression between the wortmannin + D1 + D2 and the D1 + D2 groups. CONCLUSION: PARP-1 and p53 might play important roles in AR induced by LDR.

Effect of 2-TeCD on the expression of adhesion molecules in human umbilical vein endothelial cells under the stimulation of tumor necrosis factor-alpha.

Adhesion molecules play an important role in the pathogenesis of atherogenesis. They are expressed on endothelial cells surface in response to various inflammatory stimuli. In this paper, we examined the effect of 2-tellurium-bridged beta-cyclodextrin (2-TeCD), a GPx mimic, on the expression of adhesion molecules in human umbilical vein endothelial cells (HUVECs) under tumor necrosis factor-alpha (TNF-alpha) stimulation. Experimental results indicated that 2-TeCD suppressed the TNF-alpha-induced the expression of vascular adhesion molecule-1 (VCAM-1) and intercellular cell adhesion molecule-1 (ICAM-1) on HUVECs surface in a dose-dependent manner. 2-TeCD also reduced the level of mRNA expression of VCAM-1 and ICAM-1. Furthermore, 2-TeCD inhibited THP-1 monocyte adhesion to HUVECs stimulated by TNF-alpha. Nuclear factor-kappaB (NF-kappaB) could regulate transcription of VCAM-1 and ICAM-1 genes. Western blot analysis showed that 2-TeCD inhibited the translocation of the p65 subunit of NF-kappaB into the nucleus. In short, these results indicated that 2-TeCD inhibits TNF-alpha-stimulated VCAM-1 and ICAM-1 expression in HUVECs partly due to suppressing translocation of NF-kappaB.

Effects of corticosterone, cAMP, cGMP, Ca2+, and protein kinase C on apoptosis of mouse thymocytes induced by X-ray irradiation.

OBJECTIVE: To observe the effects of signal factors of corticosterone (CS), cAMP, cGMP, Ca2+ andprotein kinase C (PKC) on lymphocyte apoptosis in mouse thymus induced by X-rays of 4 Gy in vitro. METHODS: The DNA lytic rate for thymocytes was measured by fluorospectrophotometry. RESULTS: The DNA lyric rate for thymocytes 4-8 hours after irradiation with 2-8 Gy was significantly higher than that in the control (P<0.01). As compared with the control, the DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.05-0.4 microg/mL ionomycin (Iono, P<0.05 or P<0.01) or 0.05-0.4 ng/mL phorbol myristate acetate (PMA, P<0.05 or P<0.01), respectively, was significantly increased, while the rate for thymocytes treated with 50 ng/mL cGMP was not significantly increased. The DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.2 and 0.4 microg/mL Iono (P<0.05), and 0.2 and 0.4 ng/mL PMA (P<0.05) plus 4-Gy irradiation, respectively, was significantly higher than that treated with single 4-Gy irradiation, while the rate for thymocytes treated with 50 ng/mL cGMP plus 4-Gy irradiation was not increased. When both 0.4 microg/mL Iono and 0.4 ng/mL PMA acted on the thymocytes, the DNA lytic rate for thymocytes was significantly higher than that in the control (P<0.01), the DNA lytic rate for thymocytes treated with both 0.4 microg/mL Iono and 0.4 ng/mL PMA plus 4-Gy irradiation was significantly higher than that treated with single 4-Gy irradiation (P<0.05), but was not significantly higher than that treated with 0.4 microg/mL Iono plus 4-Gy irradiation or 0.4 ng/mL PMA plus 4-Gy irradiation. CONCLUSION: CS, cAMP, Ca2+, and PKC signal factors can promote thymocyte apoptosis induced by larger dose X-rays.

Apoptotic cell death induced by low-dose radiation in male germ cells: hormesis and adaptation.

Biological effects of low-dose radiation (LDR) in somatic cells have captured the interest of radiobiologists for the last two decades. Apoptosis of germ cells is required for normal spermatogenesis and often occurs through highly conserved events, including the transfer of vital cellular materials to the growing gametes following death of neighboring cells. Apoptosis of germ cells also functions in diverse processes, including removal of abnormal or superfluous cells at specific checkpoints, establishment of caste differentiation, and individualization of gametes. Moreover, germ cells are very sensitive to radiation-induced genomic and cytological effects. Therefore, induction of germ-cell apoptosis has been observed in the testis of animals exposed to both high-dose radiation (HDR) and LDR. Exposure of male germ cells to LDR induces a stimulating effect, while exposure to HDR causes an inhibitory effect on the metabolism, antioxidant capacity, and proliferation and maturation of cells, a phenomenon termed hormesis. Preexposure to LDR also protects cells from subsequently HDR-induced genomic and cytological effects, a phenomenon termed adaptive response. This review describes the features of male germ-cell apoptosis. It reviews the evidence that LDR induces the hormesis and adaptive responses in the male germ cells in terms of apoptosis. This review also discusses the possible effects of LDR-induced apoptotic hormesis and adaptive response on the modulation of inheritable genomic damage caused by subsequent radiation exposure to male germ cells.

The macrophage heterogeneity: difference between mouse peritoneal exudate and splenic F4/80+ macrophages.

Macrophages isolated from various tissues manifest differences in cell shape, the expression of surface markers, as well as metabolic and functional activities. However, the heterogeneity of macrophages expressing the same marker in different tissues has not been fully addressed. In the present study, mouse F4/80+ peritoneal exudate macrophages (PEMs) and splenic macrophages (SPMs) appeared similar in most respects. But the percentages of cells expressing CD80, CD40, MHC-II, TLR2, or TLR4, but not CD11c, CD54, or CD23, in freshly isolated F4/80+ SPMs were significantly higher than those in PEMs, whereas the levels of CD86+ cells in F4/80+ SPMs were markedly lower than those in PEMs. After lipopolysaccharide (LPS) stimulation, F4/80+ SPMs expressed significantly higher levels of CD86, CD40, or MHC-II than F4/80+ PEMs, but not CD11c, CD80, CD54, or CD23. F4/80+ SPMs had remarkably lower non-opsonic phagocytotic capacity against chicken RBCs or allo-T cells than PEMs as determined by two-photon microscopes and flow cytometry. SPMs produced markedly more NO than PEMs when cultured with LPS or allo-T cells. Furthermore, SPMs exhibited stronger immunogenicity than PEMs, as determined by the ability to stimulate T cell proliferation, delayed type hypersensitivity, and IFN-gamma production. The data showed the differences between SPMs and PEMs with regard to the phenotypes, phagocytosis, and immunogenicity, which may offer important information for us to better understand the distinguished immune responses of macrophages in spleens and the peritoneal cavity.

Toll-like receptors and graft rejection.

Innate immunity plays a role in fighting against invading microorganisms. Emerging evidence suggests that in addition to responding to pathogen-associated molecular patterns of microorganisms, Toll-like receptors (TLRs) can be activated by endogenous ligands expressed by mammalian cells. Clinical and laboratory studies have shown that TLRs may participate in organ graft rejection and transplant immune tolerance, which are briefly reviewed in the present manuscript.

Effect of low-level radiation on the death of male germ cells.

Hormetic and adaptive responses induced by low-level radiation in hematopoietic and immune systems have been observed, as shown by stimulatory effects on cell growth and resistance to subsequent radiation-induced cytogenetic damage. However, in terms of cell death by apoptosis, the effects of low-level radiation are controversial: Some studies showed decreased apoptosis in response to low-level radiation while others showed increased apoptosis. This controversy may be related to the radiation doses or dose rates and also, more importantly, to the cell types. Testes are one of the most radiosensitive organs. The loss of male germ cells after exposure to ionizing radiation has been attributed to apoptosis. In the present study, the effects of low-level radiation at doses up to 200 mGy on mouse male germ cells in terms of apoptosis and the expression of apoptosis-related proteins were examined at different times after whole-body exposure of mice to low-level radiation. In addition, the effect of pre-exposure to low-level radiation on subsequent cell death induced by high doses of radiation was examined to explore the possibility of low-level radiation-induced adaptive response. The results showed that low-level radiation in the dose range of 25-200 mGy induced significant increases in apoptosis in both spermatogonia and spermatocytes, with the maximal effect at 75 mGy. The increased apoptosis is most likely associated with Trp53 protein expression. Furthermore, 75 mGy low-level radiation given pre-irradiation led to an adaptive response of seminiferous germ cells to subsequent high-level radiation-induced apoptosis. These results suggest that low-level radiation induces increased apoptosis in male germ cells but also induces a significant adaptive response that decreases cell death after a subsequent high-dose irradiation.

[Changes of cycle and apoptosis of spermatogenic cells and antioxidant capacity in male rats with diabetes mellitus].

OBJECTIVE: To explore the changes of cycle and apoptosis of spermatogenic cells and antioxidant capacity of the serum and testis in male rats with diabetes mellitus. METHODS: Thirty male rats were divided into two groups, 10 for normal control and 20 for the diabetes group. The rats were injected intraperitoneally with streptozocin (TZ) to develop diabetes, and 12 weeks later, their survival rate and testis weight were recorded. The percentage of G0/G, S and G2/M phases and apoptosis in spermatogenic cells were measured with flow cytometry (FCM). Malondialdehyde (MDA) and nitric oxide (NO) levels, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and NO synthase (NOS) activities in the serum and testis were measured with thiobarbituric acid reactive substances (TBARs), nitric acid reoxidized enzyme, xanthine oxidative enzyme, 5,5 Dithiobis (2,2 nitrobenzoate) (TNB) and visible light photometer methods, respectively. RESULTS: Twelve weeks after the male rats got diabetes, their survival rate, body weight and testis weight were significantly lower (p < 0.05), and the percentages of G0/G1 phases and apoptotic spermatogenic cells were obviously higher (P < 0.05) than the normal control. At the same time, the percentage of S and G2/M phases spermatogenic cells decreased. So the spermatogenic cells were arrested in G0/G1 phase. In the diabetic rat serum and testis, especially in the testis, MDA levels were distinctly higher and SOD activities were significantly lower than those in the control. Serum GSH-Px activities of the diabetic rats were significantly lower (p < 0.05), while testis GSH-Px activities were significantly higher than those in control group (P < 0.01). NO contents in the serum and testis of the diabetic rats (P < 0.01) increased significantly, particularly the former, while NOS activities in the serum decreased significantly as compared with the control (P < 0.5). CONCLUSION: The increase in testis and serum MDA levels and NO contents and the decrease in the antioxidant enzyme activity of the diabetic rats may be relevant to spermatogenic disorder caused by the increase of G0/G1 phases arrest and spermatogenic cells apoptosis.