Effect of DC vaccine sensitized with CpG ODN combined with tumor antigen on murine melanoma / 药学学报

The potential application of dendritic cells (DC) sensitized with cytosine-phosphoric acid-guanine (CpG) oligodeoxynucleotide (ODN) and tumor antigen as a vaccine against murine melanoma was investigated with freshly isolated mouse bone marrow-derived dendritic cells. For the DC vaccine preparation, DC were sensitized with the B16 tumor antigen and CpG ODN was used to promote further maturation of the DC. The immunogenic activity of the vaccine was evaluated in vitro by determining the proliferation of T lymphocytes and the killing effect of cytotoxic T lymphocytes (CTL) on B16 tumor cells. The DC vaccine was injected intraperitoneally and tumor inhibition in mice bearing B16 xenografts was examined. All mice were cared for under an approved SIMM Institutional Animal Care and Use Committee (IACUC) protocol. In vitro, this DC vaccine promoted the proliferation of T lymphocytes and showed a potent killing effect on the target B16 cells. In vivo experiments showed that after treatment or pre-immunization both the tumor volume and weight were significantly decreased. The DC vaccine with CpG ODN and tumor antigen exhibited an inhibitory effect against melanoma, providing a potential method for melanoma cancer treatment.

The effect of microRNA-145 on immune-inflammatory response of foam cells by targeting CD40 / 天津医药

Objective To investigate the effect of miRNA-145 (miR-145) on immuno-inflammatory reaction of foam cells by targeting CD40. Methods Mouse macrophage cell line RAW 264.7 cells cultured in vitro were randomly divided into model group (non-transfected), miR-145 mimics group (transfected miR-145 mimics), miR-145 inhibitor group (transfected miR-145 inhibitor) and silencing CD40 sequence group (transfected siCD40). Then oxidized low density lipoprotein (ox-LDL) was used to stimulate for 24 h to establish immune inflammatory damage cell model. Quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot assay were used to detect the levels of CD 40 mRNA and protein of each group. ELISA was used to detect the levels of inflammatory factors interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) -α in cell supernatant. Results Compared with model group, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all significantly decreased in miR-145 mimics group (P<0.01). After transfected with miR-145 inhibitor, the above indexes were all significantly increased than those of model group and miR-145 mimics group (P<0.01). After transfected with CD40 siRNA, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all obviously decreased compared with those of miR-145 inhibitor group (P<0.01). Conclusion MiR-145 can regulate the immune inflammatory process of foam cells through the target gene CD40, inhibit the activation of CD40/CD40L signaling pathway and inhibit inflammatory response.

microRNA-155 inhibits immuno-inflammatory reaction of foam cells by targeting MyD88 / 天津医药

Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.

microRNA-155 inhibits immuno-inflammatory reaction of foam cells by targeting MyD88 / 天津医药

Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.

Protective effect of lycopene on human spermatozoa during cryopreservation and its mechanism / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism.</p><p><b>METHODS</b>Semen samples were collected from 25 volunteers, each sample equally divided into four parts to be cryopreserved with cryoprotectant only (Ly0 control) or cryoprotectant + lycopene at the concentrations of 2 (Ly2), 5 (Ly5), and 10 µmol/L (Ly10), respectively. Before and after thawing, the semen samples were subjected to computer-assisted semen analysis ( CASA) for sperm kinematics, flow cytometry for sperm apoptosis, thiobarbituric acid assay for malondialdehyde (MDA) concentration, and JC-1 fluorescent staining for the sperm mitochondrial membrane potential (MMP).</p><p><b>RESULTS</b>After cryopreservation, sperm motility was markedly decreased in all the groups (P < 0.01). The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control ([25.68 ± 4.36]% vs [33.26 ± 4.78]%, P < 0.05), while sperm MMP remarkably higher in the former than in the latter ([66.18 ± 14.23]% vs [55.24 ± 12.31]%, P < 0.05). The Ly2, Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control (P > 0.05).</p><p><b>CONCLUSION</b>Addition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process, attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane, and improve the anti-apoptosis ability of sperm.</p>

Expressions of differential proteins in the expressed prostatic secretion of patients with III A chronic prostatitis / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the expressions of differential proteins in the expressed prostatic secretion (EPS) of patients with III A chronic prostatitis and healthy men.</p><p><b>METHODS</b>We collected EPS samples from 35 patients with III A chronic prostatitis and 18 age-matched healthy men, and detected the differentially expressed proteins in EPS by MALDI-TOF/MS. Based on the data obtained, we conducted a statistical analysis on the mass-to-charge (m/z) ratios of different proteins and a retrieval analysis on the relevant proteins using the protein database.</p><p><b>RESULTS</b>In the comparative studies of the III A chronic prostatitis patients and healthy men, 5 proteins were detected as at least 2-fold differentially expressed, which were probably brevinin-2Eg, big endothelin-1, alpha-defensin 15, beta-defensin 134 and prostatic steroid-binding protein C2. The m/z ratios were significantly up-regulated in 3 372, 3 487, 425 and 5 325 Da proteins (P < 0.01) and down-regulated in 10631Da (P < 0.01).</p><p><b>CONCLUSION</b>Proteins are differentially expressed in the EPS of III A chronic prostatitis patients and healthy men, and these proteins may be significantly correlated with the development and progression of III A chronic prostatitis.</p>

Effects of Mycobacterium vaccae vaccine on immunological function in elderly patients with stable chronic obstructive pulmonary disease / 中华老年医学杂志

Objective To investigate the effects of Mycobacterium vaccae vaccine on immunological function and clinical character in elderly patients with stable chronic obstructive pulmonary disease (COPD). MethodsA total of 100 elderly patients with stable COPD were randomly divided into immunotherapy group (group A, n= 50) and non-immunotherapy group (group B, n= 50), and normal control group (group C, n = 50). The levels of peripheral blood T-lymphocyte subsets (CD3+ , CD4+, CD8+ , CD4+/CD8+ ratio), natural killer cells (NK cells), immunoglobulins (IgG, IgA, IgM) and cytokines (IL-6, IL-8, TNF-a) were measured respectively before and after therapy. Group A and B were followed up for 1 year, then the times of acute outbreak and hospitalization of patients in the two groups were also compared. Results The levels of CD4 + ,CD4+/CD8+ ratio and NK cells in group A, B were significantly lower before therapy (P＜0. 05～0. 01＝, and the levels of IL-6, IL -8, TNF-a and IgA were significantly higher than in group C (P＜0. 01＝. After treatment with Mycobacterium vaccae vaccine in group A, the levels of CD4+ , CD4+/CD8+ ratio and NK cells were significantly higher (P＜0. 05-0. 01＝ and IL-6, IL-8, TNF-a and IgA were significantly lower than before treatment (all P＜0. 01＝. These levels showed no significant changes in group B after treatment (P＞0. 05). After 1-year follow-up, the times of acute outbreak and hospitalization on patients were statistically lower in group A than in group B (P＜ 0. 01 ).ConclusionsMycobacterium vaccae vaccine can improve cellular immunity function and reduce the times of acute outbreak and hospitalization in patients with stable COPD, so it has a higher clinical application value.

Effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease (AD).</p><p><b>METHODS</b>Thirty-two male SD rats (250-300 g) were randomized into 4 groups (n=8), including the normal control group without any treatment, AD model group with right intracerebroventricular injection of 2 µl Aβ25-35 (10 mg/m1), sham-operated group with right intracerebroventricular injection of 2 µl normal saline, and AD+ rAAV-HIF-1α group with right intracerebroventricular injection of 10 µl rAAV-HIF-1a (1×10¹² v.g./m1) one week after Aβ25-35 injection. The rats were sacrificed to detect the expression of HIF-1α and apoptosis of hippocampal neurons 5 weeks after Aβ25-35 or saline injection.</p><p><b>RESULTS</b>Western blotting showed that the expression of HIF-1α was significantly higher in AD+rAAV-HIF-1α group (451.59±34.39) than in normal control group (229.05±41.28) and sham-operated group (216.29±37.08) (P<0.05) without significant difference between the latter two groups. The apoptotic ratio of the hippocampal neurons was significantly higher in AD model group ([19.49±2.59]%) than in normal control group ([5.41±0.75]%) and sham-operated group ([5.28±0.66]%) in (P<0.05), and intracerebroventricular injection of rAAV-HIF-1α resulted in a significant reduction of the apoptotic ratio in the AD rats ([12.07±2.06]%) (P<0.05).</p><p><b>CONCLUSION</b>Intracerebroventricular injection of rAAV-HIF-1α can inhibit hippocampal neuronal apoptosis in the rat model of AD.</p>

The clinical application of 320-slice Computed Tomography (CT) hepatic artery images in patients with liver transplantation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the clinical significance of 320-slice CT hepatic artery images in patients with liver transplantation.</p><p><b>METHODS</b>A total of 58 patients underwent CT scanning by 320-slice scanner after liver transplantation. They were divided into 2 groups according to the concentration of contrast media as follows: Group A (27 cases, 350 mgI/ml iopromide), Group B (31 cases, 370 mgI/ml iopromide). Contrast medium was infused at 6 ml/s, with a total dose of 50 ml. Images were generated by dynamic volume scanning and were processed by 4D digital subtraction angiography (DSA) imaging software. The time-density curve (TDC) of the hepatic artery was delineated. The time to peak, peak contrast enhancement were recorded. The physiological parameters such as body weight and height were analyzed.</p><p><b>RESULTS</b>(1) There were no differences in clinical parameters such as age, sex, height, weight, or BMI between groups. The time to peak of hepatic artery of group A and B was (19.71+/-3.11) s and (20.06+/-3.67) s, and had no significant difference. The maximum peak enhancement of hepatic artery in groups B was higher than that group A (P < 0.05). (2) 4D DSA revealed hepatic artery pseudo-aneurysm (n = 2), and hepatic artery mild stenosis (n = 13), moderate stenosis (n = 5), severe stenosis (n = 9) and occlusion (n = 1), segmental moderate and severe stenosis (n = 4), and compensatory circulation with hepatic artery severe stenosis and occlusion (n = 6). hepatoportal arteriovenous fistulas (HPAVF, n = 12), donor-recipient hepatic artery mismatch (n = 3). Hepatic arterial branch are decreased and opened in 15 cases and 8 cases.</p><p><b>CONCLUSION</b>320-slice CT hepatic artery images is safe, noninvasive, and accurate technique to evaluate hepatic arterial complications after liver transplantation.</p>

Intrathecal injection of MK-801 inhibited the NOS activity and NO content of hippocampus in rat during the process of formalin-induced inflammatory pain / 中国应用生理学杂志

<p><b>AIM</b>To study the effect of intrathecal injection of MK-801, a NMDA receptor antagonist, on the NOS activity and NO content of hippocampus in rat during the process of formalin-induced inflammatory pain as well as the pain behavior of rat.</p><p><b>METHODS</b>The degree of pain was determined by observing the time of licking and biting the injected paw. NOS expression in the hippocampus was determined by using NADPH-d histochemical staining. NO content of hippocampus was determined by assaying NO3; and NO2.</p><p><b>RESULTS</b>Subcutaneous injection of formalin elicited a characteristic pain behavioural response consisting of licking and biting the injected paw, etc. Intrathecal injection of MK-801 could shorten obviously the time of licking and biting representing pain behavioural response in phase 2. It is suggested that intrathecal injection of MK-801 could block the pain behavioural response induced by formalin (P < 0.05). The number and staining degree of NADPH-d positive neurons in formalin group significantly increased at 12 h after the formalin injection in CA1, CA2-3 and DG of hippocampus compared with control group as well as NO content, however, the number and staining degree of NADPH-d positive neurons in formalin + MK-801 group significantly decreased in contrast to those of formalin 12 h group as well as the NO content (P < 0.01).</p><p><b>CONCLUSION</b>Intrathecal injection of NMDA receptor antagonist MK-801 could inhibit the NOS activity and NO production in hippocampus of rat, which showed the increase of hippocampal NO production was mainly induced by the peripheral nociceptive information input.</p>

Oxidative stress status in patients with chronic obstructive pulmonary disease and its relation to glucocorticoid receptor levels / 南方医科大学学报

<p><b>OBJECTIVE</b>To study changes in the levels of systematic and airway local oxidative stress in patients in different stages of chronic obstructive pulmonary diseases (COPD), and explore the association between oxidative stress and glucocorticoid receptor (GR) level in the peripheral blood leukocytes.</p><p><b>METHODS</b>The levels of malonaldehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in induced sputum and plasma, as well as GR levels in peripheral blood leukocytes and plasma levels of cortisol and adrenocorticotrophic hormone (ACTH), were examined in 33 patients with acute exacerbations of COPD (AECOPD, group A), 27 with stable COPD (group B), and 28 healthy volunteers (including 15 smokers as group C, and 15 nonsmokers as group D).</p><p><b>RESULTS</b>MDA level in induced sputum and plasma decreased, whereas the levels of GSH, SOD and GSH-PX increased significantly in the order of groups A, B, C, and D (P<0.05). The activity of SOD in induced sputum and plasma were significantly lower in group C than in group D. No significant difference was noted in the other oxidative stress indices between groups C and D (P>0.05). The plasma levels of cortisol and ACTH showed no significant difference between the 4 groups, while the GR level in peripheral blood leukocytes increased significantly in the order of groups A, B, C and D (1565-/+719, 2069-/+488, 2739-/+926, and 4793 -/+1415 U, respectively, P<0.05). After controlling for the factor of smoking status, the plasma and sputum SOD activity were both positively correlated to GR, with the partial correlation coefficient of 0.512 and 0.564, respectively (P<0.001).</p><p><b>CONCLUSION</b>Patients in different stages of COPD, especially those with AECOPD, may sustain systematic and local oxidation and anti-oxidation imbalance. Decreased SOD activity may contribute to GR level decrement in peripheral blood leukocytes in these patients.</p>

Formalin-induced pain enhanced nitric oxide synthase expression and nitric oxide production in the rat hippocampus / 中国应用生理学杂志

<p><b>AIM</b>To observe the changes of nitric oxide synthase (NOS) activity and nitric oxide (NO) content of hippocampus including their time course and region distribution character in rat during the process of formalin-induced inflammatory pain as well as the pain behavior of rat.</p><p><b>METHODS</b>The pain threshold (PT) was determined by radiant heat-induced tail flick test. NOS expression in the hippocampus was determined by using NADPH-d histochemical staining. NO production in hippocampus was determined by assaying NO3- and NO2-.</p><p><b>RESULTS</b>Subcutaneous injection of formalin elicited nociceptive behavioural response and led to decrease in PT of rat. The number and staining degree of NADPH-d positive neurons began to increase at 6 h after the formalin injection in CA1, CA2 - 3 and DG of hippocampus as well as NO content, which increased most obviously at 12 h and returned to control level at 48 h.</p><p><b>CONCLUSION</b>Formalin-induced inflammatory pain could induce the elevation of NOS activity in CA1, CA2 - 3 and DG of hippocampus with a certain time course, which further led to a increase of NO production in hippocampus.</p>

Effects of scopolamine on morphine-dependence in mice and the relationship between the effect and hippocampus intracellular calcium / 中国应用生理学杂志

<p><b>AIM</b>To observe the effect and mechanism of scopolamine on morphine(Mor)-induced mice dependence.</p><p><b>METHODS</b>The Mor-dependent mice model was established by intraperitoneal (ip) administered Mor for seven days. Pain threshold, times of jump and hippocampus intracellular free calcium ion concentration ([Ca2+]i) were determined by the heat plate test, naloxone (Nal)-precipitated jumping response and flow cytometry, respectively.</p><p><b>RESULTS</b>The pain threshold of Mor-dependent mice decreased significantly while there was a marked increase in times of jump, the rate of jumping animals and hippocampus [Ca2+]i. Co-administered scopolamine, the pain threshold of Mor-dependent mice increased significantly; the number of jump, the rate of jumping animals and hippocampus [Ca2+]i all decreased significantly.</p><p><b>CONCLUSION</b>Scopolamine could antagonize the Mor-induced mice dependence, which could be related to decreasing the levels of brain intracellular free calcium.</p>

Protein kinase C agonist phorbol 12-Myristate-Acetate induces nociception and enhances nitric oxide production in the spinal cord of rats / 中国应用生理学杂志

<p><b>AIM</b>To determine the involvement of NO signal pathway in the development of hyperalgesia induced by activation of protein kinase C (PKC ), nociceptive responses and nitric oxide synthase(NOS) expression and nitric oxide (NO) content in the spinal cord were observed after administration of Phorbol 12-Myristate-Acetate (PMA), a PKC agonist, in rats.</p><p><b>METHODS</b>Nociceptive response was observed by behavioral approach. Pain threshold was assayed using thermal tail-flick test. NADPH-d histochemistry was used to investigate the changes of NOS expression. Nitrate/nitrite (NO3-/NO2-) was assayed to represent NO content of lumbar enlargement of spinal cord.</p><p><b>RESULTS</b>Nociceptive response was induced and pain threshold decreased after intrathecal injection of PMA. The number of NADPH-d positive cells increased significantly in the superficial layer of the spinal cord dorsal horn (Laminae I - II ) and the grey matter surrounding the central canal (Laminae X), and the reactive degree of NADPH-d positive soma and processes and NO content of the lumbar enlargement of the spinal cord increased significantly after intrathecal injection of PMA. Pretreatment of PKC inhibitor chelerythrine chloride blocked the changes induced by PMA.</p><p><b>CONCLUSION</b>The activation of PKC in the spinal cord neurons might induce spontaneous nociceptive responses and hyperalgesia in rats, as well as promote NOS expression and NO production, suggesting that increase in NO production is one of mechanisms of hyperalgesia induced by activation of PKC.</p>