Reversal of ciprofloxacin-induced testosterone reduction by probiotic microbes in mouse testes.

CPFX is a highly effective antibiotic, but it has been reported to significantly impair both testicular function and structure in rats. In this study, we assessed reversal of CPFX-induced variation in mice testicular structure and testosterone synthesis by probiotic microbes in the infected model and normal model. We detected testicular weight, testicular structure and Leydig cell variables in numbers. We detected the levels of serum testosterone and steroidogenic enzymes, as well as DBC1, Sirt1, NF-κB, and related redox state and inflammatory response in the testes. The results showed that probiotic microbes had significantly elevated serum testosterone levels and steroidogenic enzymes, higher Sirt1, anti-oxidative enzymes and anti-inflammatory cytokine expression, and lower NF-κB, DBC1, oxidative damage, pro-inflammatory cytokine expression. The results suggest that the testis-protective, antiinflammatory and antioxidation effects of probiotics largely resulted from its ability to decrease oxidative stress and preserve antioxidant activity by stabilizing antioxidant defense systems, reducing oxidative damage and inflammatory response.

Metabonomic analysis of quercetin against the toxicity of chronic exposure to a mixture of four organophosphate pesticides in rat plasma.

1. A metabonomics approach was performed to investigate the effect of quercetin on the toxicity of chronic exposure to a mixture of four organophosphate pesticides (OPs) at their corresponding no-observed-adverse-effect level (NOAEL). The rats were divided into six groups (n = 10/group): control, two different doses of quercetin, OPs mixture and different doses of quercetin plus OPs mixture-treated groups. 2. Nine metabolites, including two quercetin metabolites and seven endogenous metabolites were identified in plasma. The intensities of metabolites significantly changed in the OP mixture-treated group compared with the control group (p < 0.01), such as lysoPE (16:0/0:0), lysoPC (17:0/0:0), lysoPC (15:0/0:0) and 4-pyridoxic acid, significantly increased; by contrast, the intensities of arachidonic acid and citric acid significantly decreased. Anomalous intensity changes in aforementioned metabolites were alleviated in the OP mixture plus 50 mg/kgcbw/d quercetin-treated group compared with the OP mixture-treated group (p < 0.05). 3. The results indicated that quercetin elicited partial protective effects against the toxicity induced by a mixture of OPs, which include regulation of lipid metabolism, improvement of tricarboxylic acid (TCA) cycle disorders, enhancement of antioxidant defence system to protect the liver.

Enhancing the therapeutic efficacy of NK cells in the treatment of ovarian cancer (Review).

Ovarian cancer is a prevalent gynecological malignancy associated with a high mortality rate and a low 5­year survival rate. Typically, >70% of patients present with an advanced stage of the disease, resulting in a high number of ovarian cancer­associated deaths worldwide. Over the past decade, adoptive cellular immunotherapy has been investigated in clinical trials, and the results have led to the increased use in cancer treatment. Natural killer (NK) cells are cytotoxic lymphoid cells that recognize and lyse transformed cells, thereby impeding tumor growth. Thus, NK cells exhibit potential as a form of immunotherapy in the treatment of cancer. However, some patients with ovarian cancer treated with NK cells have experienced unsatisfactory outcomes. Therefore, further optimization of NK cells is required to increase the number of patients achieving long­term remission. In the present review article, studies focusing on improving NK cell function were systematically summarized, and innovative strategies that augment the anticancer properties of NK cells were proposed.

Percutaneous epididymal sperm aspiration and short time insemination in the treatment of men with obstructive azoospermia.

OBJECTIVE: To study the efficacy of percutaneous epididymal sperm aspiration (PESA) in combination with short time insemination to treat infertile men with obstructive azoospermia (OA). DESIGN: Paired randomized controlled trial in which each couple's cohort of oocytes was divided into two equal groups. SETTING: Center for reproductive care. PATIENTS: Twenty men with OA. INTERVENTIONS: Motile spermatozoa were collected using PESA. Half of the oocytes were used for intracytoplasmic sperm injection (ICSI). The rest were inseminated briefly with PESA sperm in vitro fertilization (IVF). After 4-5 h, the remaining cumulus cells were removed mechanically for second polar body observation to decide whether to apply "rescue" ICSI (RE-ICSI). MAIN OUTCOME MEASURES: Rates of oocyte maturation, fertilization, cleavage, and good quality embryos. Numbers of available embryos and good quality embryos were compared between PESA-IVF (using a short incubation protocol + rescue ICSI) group and PESA-ICSI group. RESULTS: In the short time insemination group, cumulus cells were dispersed by PESA spermatozoa. No second polar bodies were found, so RE-ICSI was done. PESA-IVF + RE-ICSI and PESA-ICSI outcomes were comparable in terms of fertilization rates, 2PN cleavage rate and good quality embryo rates with no statistically significant differences. CONCLUSIONS: PESA sperm without centrifugation could disperse the cumulus cells but were infertile and therefore could substitute for synthetic hyaluronidase. The outcomes of PESA-IVF with rescue ICSI were equivalent to PESA-ICSI. Using spermatozoa obtained by PESA and IVF before RE-ICIS is a viable treatment for men with OA.

[Effect of Heshouwuyin on the expression of Cox7a2 protein in testis tissue of exercised-induced fatigue rat].

OBJECTIVE: To explore the effect of Heshouwuyin on the expression of cytochrome C oxidase7a2 (Cox7a2) in testis tissue of rats with exercised-induced fatigue. METHODS: Fifty SD rats were divided into normal control group (A group), Heshouwuyin administered normal group (B group), model control group (C group), Heshouwuyin treated group (D group) and Heshouwuyin prevented group (E group) randomly with 10 rats for each. The exercise-induced fatigue models in rats of C, D, E groups were established. The rats in D group were treated with Heshouwuyin [20 g/(kg x d), contained crude drug 9.6 g/mL] for 60 days (during the 42 days of modeling and after the 18 days of modeling). The rats in E group were also treated with Heshouwuyin for 60 days (but before the 18 days of modeling and during the 42 days of modeling). Beckmancoulter Unicel Dxl 800 was used to detect the level of serum testosterone, according to the manufacture's instructions. Western blot and RT-PCR were used to observe the differential expression of Cox7a2. RESULTS: The level of serum testosterone in C group was decreased compared with A group (P < 0.05), which implied the success of modeling. Compared with group A, the level of serum testosterone in B, D, E groups were increased (P < 0.05). Cox7a2 protein was expressed mainly in leydig cell and spermatocyte. Compared with A,B, D, E groups, the expression of Cox7a2 protein and mRNA in C group increased (P < 0.05), and there no significant difference was observed between group A and B, as well as group D and E. CONCLUSION: The expression of Cox7a2 was down-regulated by Heshouwuyin.

Association between rs735482 polymorphism and risk of cancer: A meta-analysis of 10 case-control studies.

Several studies have inspected the relationship between rs735482 polymorphism and the risk of some human cancers, but the findings remain controversial. We designed this meta-analysis to validate the association between rs735482 polymorphism and cancer risk. All articles were published before September 1, 2018 and searched in Pubmed, Embase, Web of Science, China National Knowledge Infrastructure, WangFang, and Chinese BioMedical databases, STATA 12.0 software was used for statistical analysis, which provides reasonable data and technical support for this article. A total of 10 studies were included in the meta-analysis, including 2652 cancer cases and 3536 rs735482 polymorphic controls. Data were directly extracted from these studies and odds ratios with 95% confidence intervals were computed to estimate the strength of the association. By pooling all eligible studies, the rs735482 polymorphism showed no significant association with susceptibility of several cancers in all the five genetic models (the allelic model: OR = 1.019, 95% CI: 0.916-1.134, P = .731). In addition, another adjusted OR data showed a significant increased risk between the rs735482 and susceptibility of several cancers (the codominant model BB vs AA: OR = 1.353, 95% CI: 1.033-1.774, P = .028) and the stratification analysis by ethnicity indicated the rs735482 is associated with an increased risk of cancer in Chinese group (BB vs AA, OR = 1.391, 95% CI = 1.054-1.837, P = .020; AB+BB vs AA OR = 1.253, 95% CI = 1.011-1.551, P = .039). However, the ERCC1 rs735482 is associated with a decreased risk of cancer in Italian group (AB vs AA, OR = 0.600, 95% CI = 0.402-0.859, P = .012; AB + BB vs AA, OR = 0.620, 95% CI = 0.424-0.908, P = .014). The results of this meta-analysis do not support the association between rs735482 polymorphism and cancer risk. But stratified analysis showed that rs735482 significantly increased the risk of cancer in Chinese while decreased the risk of cancer in Italian. Because of the limited number of samples, larger and well-designed researches are needed to estimate this association in detail.

Author Correction: The roles of p38 MAPK → COX2 and NF-κB → COX2 signal pathways in age-related testosterone reduction.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Repeated exposure to the irrigative wastewater in Shijiazhuang induced precancerous lesion associated with cytochrome P450.

In the present study, the carcinogenic effects of the wastewater sample collected from the Dongming Canal in Shijiazhuang city were first detected by the rat medium-term liver bioassay. The experiment contained five groups: a negative control group, a DEN-alone group, 25% wastewater, 50% wastewater, and 100% wastewater. The body weight of rats decreased significantly as the dose increased. Morphologically, we also found that the damage of the hepatic lobule was more serious and the proliferation of liver cells was more obvious as the dose increased. In addition, we observed a significantly increased liver organ coefficient in rat. With the increase in dose, the damage of the hepatocytes was more serious, which was manifested in significantly elevated of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and gammaglutamyl transfer peptide enzyme (γ-GT). And, the irrigative wastewater significantly increased GST-p in the liver of rats at both the transcriptional and translational levels dose-dependently, eventually causing precancerous lesions in the liver tissues. CYP1A1 and CYP1B1 expressions in the rat liver cells at the level of transcription and translation were also significantly increased dose-dependently. Our data clearly demonstrated that the irrigative wastewater had a carcinogenetic effect that was associated with CYP1A1 and CYP1B1. The risk of carcinogenic potential to human health might be due to joint action and accumulative effects over a long period of exposure. We can also concluded that the medium-term liver bioassay could be used as an effective method for evaluating the carcinogenicity of complex water mixtures such as irrigative wastewater.

The roles of p38 MAPK → COX2 and NF-κB → COX2 signal pathways in age-related testosterone reduction.

In our study, we explored changes in the redox status and inflammatory response in the testes of the SAMP8 model of varying ages (2, 4, 8, 10 months old) compared with control mice SAMR1 by the methods of immunohistochemical staining, Western blotting, RT-PCR and Luminex multi-analyte cytokine profiling. We found that as ROS and inflammation levels increased during aging, steroidogenic enzymes (StAR and P450scc) reduced and led to the decline of testosterone production eventually. The pathways of P38 MAPK → COX2 and NF-κB → COX2 were detected by using specific inhibitors of SB203580 and Bay 11-7082 in isolated Leydig cells. These results indicated that activation of both p38 MAPK → COX2 and NF-κB → COX2 signaling pathways are functionally linked to the oxidative stress response and chronic inflammation during aging, and mediate their inhibitory effects on testosterone production.

[Expression of Sema3A and NP-1 in spinal cord and spared dorsal root ganglion after partial dorsal root rhizotomy of cat].

OBJECTIVE: To investigate the expression of semaphorin 3A (Sema3A) and neuropilin 1 (NP-1) in spinal cord and dorsal root ganglion after partial dorsal root rhizotomy. METHODS: 15 adult cats were used for this study and divided into 3 groups: normal control group, 7 d and 14 d postoperative groups (7Th day and 14th day groups) undergoing unilateral partial dorsal root rhizotomy. The L3, L5 and L6 segments of spinal cord and L6 dorsal root ganglia (DRG) in operated side were made into frozen sections. By immunohistochemistry ABC method, the sections of spinal cord were stained with specific Sema3A antibody, and L6 DRG were stained with NP-1 antibody. The mean optical density (OD) of Sema3A immunoreactivity in dorsal horn was measured and the number of NP-1 positive medium-small sized neurons in spared DRG was counted. RESULTS: After partial dorsal root rhizotomy, in L3 segment the expression of Sema3A decreased in 7th day group (0.25 +/- 0. 14) compared with that in normal group(0. 37 +/- 0.87) (P < 0.05), but kept the level along to 14th day group (0.27 +/- 0.09); in L5 segment, the expression of Sema3A decrea sed in 7th day group (0.26 +/- 0.11) (P < 0.05), and then recovered to normal level in 14th day group (0.33 +/- 0.09); in L6 segment, OD values in dorsal horn had no changes to all groups. The number of NP-1 positive medium-small sized neurons in spared DRG (30.85 +/- 10.26) was decreased in 7th day group (P < 0.05), compared with that in normal group (45.06 +/- 12.47), while increased in 14th day group (P < 0.05). CONCLUSION: Changes in the expression of Sema3A in spinal cord and the expression of NP-1 in L6 DRG after partial root rhizotomy may be involved in collateral sprouting of spared root in superficial lamina.

Ferroptosis is associated with oxygen-glucose deprivation/reoxygenation-induced Sertoli cell death.

Sertoli cell death contributes to spermatogenesis impairment, which is associated with male infertility. Testicular ischemia­reperfusion (I/R) injury induces the cell death of germ cells and Sertoli cells, whereas inhibition of cell death ameliorates acute testicular I/R damage. The aim of the present study was to investigate the mechanism of I/R stress-induced cell death in TM4 cells. Oxygen­glucose deprivation and reoxygenation (OGD/R) was demonstrated to induce I/R injury and cell death in TM4 cells. Cell death was blocked by the reactive oxygen species (ROS) inhibitor N­acetylcysteine, as well as lipid peroxidation inhibitors Liproxstatin­1 and iron chelator deferoxamine; however, inhibitors of apoptosis, necrosis or autophagy had no effect. It was also demonstrated that iron and lipid ROS levels were elevated in I/R injury and that mitochondria decreased in size and increased in membrane density, which is indicative of ferroptosis. Furthermore, the generation of lipid ROS suggests iron accumulation and glutathione (GSH) depletion. The expression of ferroportin (Fpn) protein and mRNA was decreased in TM4 cells. Notably, overexpression of Fpn inhibited ferroptosis, lipid ROS generation and iron accumulation. In addition, GSH­dependent peroxidase 4 (GPX4) was inactivated via GSH depletion following I/R injury, whereas GPX4 activation blocked I/R­induced ferroptosis by reducing lipid ROS levels. The mitogen­activated protein kinase (MAPK) pathway was also investigated in the present study; it was observed that I/R­induced ferroptosis was blocked by inhibiting p38 MAPK activation. The results of the present study demonstrate that ferroptosis is a pervasive and dynamic type of cell death induced by OGD/R injury in Sertoli cells. This may provide a novel insight into the application of cytoprotection in testicular I/R damage­induced cell loss.

FOXK1 facilitates cell proliferation through regulating the expression of p21, and promotes metastasis in ovarian cancer.

Ovarian cancer is one of the most common cancer in the world. FOX family plays essential function in multiple cancers. In our work, FOXK1 was found to up-regulate in ovarian cancer tissue samples and cell lines; moreover, the expression of FOXK1 was correlated with tumor size, metastasis and poorly prognosis. To evaluate the function of FOXK1 in ovarian cancer, we performed colony formation analysis, CCK-8 assay and cell cycle analysis to determine the effect of FOXK1 on cell proliferation and cell cycle. We found that FOXK1 obviously improved the ability of cell proliferation through promoting cell cycle. Furthermore, ChIP assay and luciferase reporter assay indicated that FOXK1 facilitated cell cycle through regulating the expression of p21, but FOXK1 had no effect on cell apoptosis. In addition, wound healing assay and transwell invasion analysis demonstrated that FOXK1 promoted migration and invasion in ovarian cancer. In conclusion, our work indicate FOXK1 plays a key function in the ovarian cancer, it promotes cell proliferation and metastasis. FOXK1 serves as a novel molecular therapy target in ovarian cancer.

Versican splice variants in human trabecular meshwork and ciliary muscle.

PURPOSE: Versican, chondroitin sulfate glycoprotein 2, is thought to play a role in regulating aqueous humor outflow and intraocular pressure via the human trabecular meshwork (HTM) of the eye. This protein was upregulated in HTM cells when they were treated with TGF-beta. There are four splice variant forms of versican (V0-V3) with different numbers of glycoaminoglycan (GAG) attachment domains. In this study, we investigated the various isoforms of versican from ocular tissues and cultured cells. METHODS: HTM and human ciliary muscle (HCM) tissues were dissected from three pairs of donors eyes with no histories of eye disease. Cultures of HTM and HCM cells were established from five donor eyes, and HTM cells were treated for 72 h with 1 ng/ml of either with hrTGF-beta1 or hrTGF-beta2. Total RNA was isolated from cells and tissues from each of the samples. Relative quantitation of gene expression of each variant was detected by real-time PCR with SYBR Green dye. RESULTS: All four variants of versican existed in each sample. In cultured HTM cells, the two variants with the largest number of GAG domains predominate. The V1 form is about 20% greater than the V0 form. There is an upregulation, particularly in the V0 form, when the cells are cultured. In tissue, the V1 form is about five fold greater than the other three. In the ocular ciliary muscle, the V1 form is the most prominent, but with this tissue, the relative amount of the V0 form did not change when cells were cultured. There was an upregulation of all splice variants in HTM cells when they were cultured with either TGF-beta1 or TGF-beta2. The increase in expression in the HTM with TGF-beta treatments were greatest with the V0 and V2 forms. CONCLUSIONS: This is the first report about the presence of various forms of versican in the anterior segment of the eye and the alterations in the mRNA patterns of these forms when cells are placed in culture. The results indicate the variants with the largest numbers of GAG attachment domains are the most prominent in the HTM and HCM. The increases in these forms that have the most GAG domains would be consistent with the increases in chondroitin sulfate reported in glaucoma.

[Changes in expression of SP in spinal lamina II following hemisected spinal cord injury in rats].

OBJECTIVE: To explore the changes in expression of Substance P (SP) in spinal lamina II at different time following hemisected spinal cord injury (hSCI). METHODS: Twenty Sprague-Dawley rats were randomly divided into intact group and hSCI group at days 3, 7 and 21. The spinal cords were hemisected between T13 and L1. The animals were sacrificed 3, 7, 21 days after operation. The L3 segments were taken out and sectioned continuously into sections (20 microm). The expression of SP was measured by immunohistochemical ABC method, and the number of SP positive varicosities in lamina II was counted. RESULTS: SP positive varicosities were observed in spinal lamina II. Compared with control group, the number of SP positive varicosities on the injured side apparently increased 3 days after hSCI, but decreased 7 days after operation, then recovered to normal by 21 days after operation. Noteworthily, the number of SP positive varicosities on the intact side was higher than that on the injured side. CONCLUSION: SP, as a neurotransmitter of nociceptive information, may be related to the process of hSCI.

[Expression of CNTF and PDGF in spared dorsal root ganglion after partial dorsal root rhizotomy].

OBJECTIVE: To investigate the expression of ciliary neurotrophic factor (CNTF) and platelet derived growth factor (PDGF) in spared dorsal root ganglion (DRG) after partial dorsal root rhizotomy. METHODS: Twenty adult cats were divided into four groups. Five cats were kept intact in the control group. Fifteen cats were subjected to bilateral root rhizotomy, and on the 3rd day, 7th day and 14th day after operation, they were sacrificed as subjects in the three experiment groups respectively (n=5 per group) and their DRGs (L6) were taken. Immunochemical ABC method was used to detect the distribution of CNTF and PDGF-immunoreactivity neurons in those DRGs. The quantitative analysis was conducted to get the numbers of CNTF, PDGF-positive total, large, and medium-small sized neurons in each group. RESULTS: CNTF, PDGF-immunoreactants were distributed in large and medium-small sized neurons. The numbers of total and medium-small sized CNTF positive neurons were noted to be decreased on the 3rd day after operation (P<0.05), and no difference 7th and 14th days was seen when compared with control (P>0.05), but the large positive neurons showed no difference. The total and medium-small sized PDGF positive neurons were found decreased apparently on the 3rd day and 7th day, but there was no difference on the 14th day as compared with normal level. Large positive neurons displayed no change at every time. CONCLUSION: Partial dorsal root rhizotomy exerts different influence on the expression of CNTF and PDGF for different neurons in spared DRG.

[Expression of Bax, Bcl-2 and caspase-3 in spared dorsal root ganglion after partial dorsal root rhizotomy].

OBJECTIVE: Investigating the expression of Bax, Bcl-2 and Caspase-3 in dorsal root ganglion after partial dorsal root rhizotomy in spared root. METHODS: Adult cats were used. The study comprised a normal control group (n=5) and three expriment groups (n=5 per group). The unilateral root rhizotomy was performed on the 15 cats of the 3 experiment groups. On the 3rd, 7th and 14th days after partial dorsal rhizotomy, the DRG (L6) of 5 cats per batch were taken and made into frozen sections and were stained under the same condition using specific Bax, Bcl-2 and Caspase-3 antibody by ABC method. The Bax, Bcl-2 and Caspase-3 positive large-, medium- and small-sized neurons in DRG were counted in the 3 experiment groups and in the normal control group. RESULTS: In the intact animals of the normal control group, the Bax, Bcl-2 and Caspase-3 immunoreactants were mainly distributed in medium, small neurons and a few large neurons. The number of Bax positive neurons in spared DRG of the 3rd day group increased apparently as compared with that of the normal control group but decreased apparently in the 14th day group and showed no significant difference when compared versus control (P>0.05). There was no significant difference between the 14th day group and normal control group. Bcl-2 expression in spared DRG displayed no difference among the four groups. The number of Caspase-3 positive neurons in spared DRG of the 3rd day group increased apparently when compared against control (P<0.05), but it was not significantly different from that of the 7th day and 14th day groups (P>0.05). CONCLUSION: Partial dorsal root rhizotomy mainly has an effect on the expressions of Bax, Bcl-2 and Caspase-3 in medium- and small-sized neurons of spared DRG, the peaks of the expressions may appear 3-7 d after the operation.

Gene and protein expression changes in human trabecular meshwork cells treated with transforming growth factor-beta.

PURPOSE: To determine the genomic and proteomic expression changes in human trabecular meshwork cells when they are treated with transforming growth factor (TGF)-beta. METHODS: Human trabecular meshwork cells from five donors were cultured for 3 days with 1 ng/mL of either TGF-beta1 or -beta2. Changes in gene expression determined with gene microarrays and alterations in protein expression detected by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) were studied in these cells after the incubation. RESULTS: With both TGF-betas, there was a substantial upregulation of genes that were related to secreted proteins or extracellular matrix. This result was consistent with pathologic changes observed in disease and with experiments on perfused trabecular meshwork. Several of the gene changes suggest that other signaling pathways, such as ErbB and Wnt, were altered. Changes in enzyme expression in the prostaglandin pathway indicated that the prostaglandins may have a different cellular profile in the presence of glaucoma. Two genes, osteoblast-specific factor 2 and corneal-derived transcript 6, which are highly expressed in the cells under normal conditions, were substantially upregulated with the TGF-betas. Proteomic analysis indicated that there was increased proteolysis of vimentin with both treatments. Tropomyosin 1alpha was increased in both gene and protein expression, suggesting alterations of the cytoskeleton by the disease. The TGF-beta1 treatment caused more robust changes than those induced by TGF-beta2. Three genes-aldose reductase, thioredoxin reductase 1, and glucose-6-phosphate 1-dehydrogenase-were identified that were downregulated in expression. These genes had decreases in protein expression with TGF-beta1 treatment but had little change in either gene or protein expression with TGF-beta2. CONCLUSIONS: Human trabecular meshwork cells can be subjected to increased levels of TGF-beta for several years as a result of glaucoma. The results indicate that changes in extracellular matrix as well as alterations in cytoskeletal proteins occur in these cells as a result of increased TGF-beta. These results are consistent with changes observed in the trabecular meshwork in glaucoma and suggest that at least some of the histologic alterations observed in the meshwork in glaucoma may be the result of increased TGF-betas.

Effects of prostaglandin analogues on human ciliary muscle and trabecular meshwork cells.

PURPOSE: To determine the effects of prostaglandin F(2alpha) analogues on gene expression of human ciliary muscle (HCM) and trabecular meshwork (HTM) cells. METHODS: Cultures of HCM and HTM cells were established from five different donors treated for 9 days with 10 microg/mL of either latanoprost (free acid) or prostaglandin F(2alpha) ethanolamide and compared with control cells. The mRNA from the cells of the five individual donors was pooled and analyzed by using gene microarrays. Gene expression changes were confirmed by either real-time PCR or relative quantitative PCR. RESULTS: Approximately 12 genes showed a twofold or greater change in expression under experimental conditions. Four of these may alter outflow. Aquaporin-1 and versican were downregulated in the HCM, whereas IGF1 and fibroleukin were upregulated in HTM. Expression levels of TNFSF10 and promelanosome-concentrating hormone also increased in the treated HTM cells. The mRNA levels for the prostaglandin FP receptor were downregulated in the ciliary muscle cells. Optineurin and alphaB-crystallin levels remained unchanged, but myocilin in the HTM cells was decreased in some samples. CONCLUSIONS: Both analogues changed gene expression similarly in either HCM or HTM cells, but the changes appeared to be cell specific, perhaps indicating that other transcription factors are influential. Outflow of aqueous humor may be increased by the prostaglandin analogues by alterations in the extracellular matrix. Other changes may influence cellular metabolism, such as the increases in IGF1, tumor necrosis factor superfamily-10 and promelanosome-concentrating hormone.

Various roles of astrocytes during recovery from repeated exposure to different doses of lipopolysaccharide.

Previous studies have demonstrated that the outcomes associated with neuroinflammation induced by intraperitoneal injection of lipopolysaccharide (LPS) at different dosages vary and either resolve or result in sepsis. The mechanisms underlying differential recoveries from varying doses of LPS are unclear. Additionally, changes in recovery involving chronic or continuous systemic inflammatory responses remain unclear. The present experiments were designed to evaluate the effects of systemic inflammation induced by repeated intraperitoneal injection of LPS at different doses on cognitive impairment. These experiments were also designed to investigate the roles of microglia and astrocytes in systemic inflammation and confirm the mechanisms that influence these processes. Kunming mice were given intraperitoneal injections of LPS at either 5mg/kg or 10mg/kg or saline for 7 consecutive days. Following the 7-day course of injections, a number of mice were kept undisturbed in their home cage for 30 days (30-day recovery), and other mice were similarly kept for 90 days (90-day recovery). The results revealed that the cognitive and physiological changes induced by 5mg/kg LPS included weight loss, impairments in spatial learning and memory, phenotypic changes in glia cells, and altered levels of pro-inflammatory cytokines; all of which were reversible. A potential recovery mechanism involves a neuroprotective function of activated astrocytes that secreted glial-derived neurotrophic factor (GDNF) following 30-day recovery. The changes induced by 10mg/kg LPS included weight loss, phenotypic changes in glia cells, and altered levels of pro-inflammatory cytokines were also reversible; however, a longer recovery was required (90 days). Although 10mg/kg LPS-induced neuroinflammation was reversible, the associated impairments in spatial learning and memory were permanent. A potential mechanism underlying permanent damage associated with 10mg/kg LPS involves the role of the activated astrocytes changing from neuroprotection to destruction, which is mediated by increased pro-inflammatory cytokines in more serious neuroinflammation.

Effects of moderate exercise over different phases on age-related physiological dysfunction in testes of SAMP8 mice.

Oxidative stress and chronic inflammation have been implicated in the testicular aging process. Different types and moderate-intensity of regular exercise may reduce age-related physiological dysfunction associated with inflammation and oxidative stress, but such effects of moderate-intensity of exercise over different phases of life in testes have not been reported. In this study, male SAMP8 mice, a senescence-accelerated strain, were maintained as sedentary (sed) or subjected to daily 15-min periods of swimming exercise between ages of 2-7 months (lifelong), 2-4 months (earlier) or 5-7 months (late). Age-related changes, including serum testosterone levels and biomarkers of inflammation and oxidative stress were analyzed at the end of the experiment. All exercise groups showed significantly greater serum testosterone levels and decreased age-related inflammation and oxidative stress compared with the sedentary group. Exercise also increased expression and activity of the nuclear factor erythroid 2-related factor (Nrf2), a transcriptional regulator of the cellular anti-oxidant system, and decreased expression and activity of nuclear factor kappa beta (NF-κB), a mediator of inflammatory molecules, in the nucleus of testicular cells. However, lifelong and earlier groups generally showed significantly better protective effects than the late group against age-related physiological dysfunction in testes. Thus, lifelong exercise and earlier phase exercise were most effective in counteracting oxidative stress and inflammation and in preserving testes function through regulation of Nrf2 and NF-κB. These results advocate the benefits of lifelong exercise and emphasize a greater protection against male aging by instituting exercise earlier rather than late in life.