The 97th Hospital of PLA
The 97th Hospital of PLA
Zhang G.,Anhui Medical University |
Yu L.,Xuzhou Medical College |
Chen Z.-Y.,Xuzhou Medical College |
Zhu J.-S.,Xuzhou Medical College |
And 4 more authors.
Brain, Behavior, and Immunity | Year: 2016
Visceral hypersensitivity is a major contributor to irritable bowel syndrome and other disorders with visceral pain. Substantial evidence has established that glial activation and neuro-glial interaction play a key role in the establishment and maintenance of visceral hypersensitivity. We recently demonstrated that activation of spinal microglial toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor κB (NF-κB) signaling facilitated the development of visceral hypersensitivity in a rat model developed by neonatal and adult colorectal distensions (CRDs). Hypothalamic paraventricular nucleus (PVN) plays a pivotal role in the pathogenesis of chronic pain. In this study, we examined the mechanism by which microglia and neurons in PVN establish and maintain visceral hypersensitivity and the involvement of TLR4 signaling. Visceral hypersensitivity was precipitated by adult colorectal distension (CRD) only in rats that experienced neonatal CRDs. Visceral hypersensitivity was associated with an increase in the expression of c-fos, corticotropin-releasing factor (CRF) protein and mRNA in PVN, which could be prevented by intra-PVN infusion of lidocaine or small interfering RNA targeting the CRF gene. These results suggest PVN CRF neurons modulate visceral hypersensitivity. Adult CRD induced an increase in the expression of Iba-1 (a microglial marker), TLR4 protein, and its downstream effectors MyD88, NF-κB, as well as proinflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) only in rats that experienced neonatal CRDs. Intra-PVN infusion of minocycline, a nonselective microglial inhibitor, attenuated the hyperactivity of TLR4 signaling cascade, microglial activation, and visceral hypersensitivity. Taken together, these data suggest that neonatal CRDs induce a glial activation in PVN. Adult CRD potentiates the glial and CRF neuronal activity, and precipitates visceral hypersensitivity and pain. TLR4 signaling and proinflammatory cytokines TNF-α and IL-1β may participate in neuro-glial interaction during the pathogenesis of visceral hypersensitivity. © 2015 Elsevier Inc.
Zhang N.-Z.,The 97th Hospital of PLA |
Bai S.,The 97th Hospital of PLA |
Cai X.-J.,Southern Medical University |
Li L.-B.,Southern Medical University
Photodiagnosis and Photodynamic Therapy | Year: 2016
Objective: To investigate the anti-tumor and immune efficacy of photodynamic immune-therapy (PIT), the combination of photodynamic therapy and dendritic cells (DC), on murine Heps hepatoma. Methods: DCs were derived from syngeneic mouse bone marrow and then labeled with DAPI in vitro. The hepatoma model was established by subcutaneous inoculation with Heps cells in one hundred and twenty-eight mice. They were then divided into four groups at random: control group, PDT group, DC group and PIT group. Tumors in the control group were injected with normal saline. Mice in the PDT group were injected with the photosensitizer Deuteporfin 24h before irradiation. Mice in the DC group were injected with DAPI labeled dendritic cells intratumorally. Mice in the PIT group were further given an injection of DCs after photoirradiation. Tumor growth and survival time were recorded after treatment. Fluorescence of tumor draining lymph nodes was evaluated under fluorescence microscope. Cytotoxic activity of splenocytes was tested by standard lactate dehydrogenase (lactate dehydrogenase, LDH) release assay. Results: (1) Tumor growth was significantly slowed down in the PDT and PIT group compared to the control group (P < 0.01). (2) The mean survival time was significantly prolonged in the PDT and PIT group. (3) The number of fluorescent cells in the draining lymph nodes from DC group was higher than that of the PIT group. (4) The anti-tumor activity of splenocytes in the PDT and PIT group was significantly higher than that of the DC and control groups (P < 0.01, P < 0.01). Conclusions: The present study suggests that PDT can inhibit tumor growth and induce anti-tumour immune response. The combination of PDT induced by Deuteporfiin and dendritic cell is capable of amplifying the antitumor immune response. © 2015 Elsevier B.V.
Li Y.,PLA Fourth Military Medical University |
Wang K.,Chinese PLA General Hospital |
Feng Y.,PLA Fourth Military Medical University |
Feng Y.,The 97th Hospital of PLA |
And 10 more authors.
Biochimica et Biophysica Acta - Molecular Basis of Disease | Year: 2014
Silent information regulator 1 (SIRT1), a class III histone deacetylase, retards aging and plays roles in cellular oxidative stress injury (OSI). However, the biological context in which SIRT1 promotes oxidative injury is not fully understood. Here, we show that SIRT1 essentially mediates hydrogen peroxide (H2O2)-induced cytotoxicity in human umbilical vein endothelial cell (HUVEC). In HUVECs, SIRT1 protein expression was significantly increased in a dose-dependent manner after H2O2 treatment, whereas the acetylation levels of the NF-κB p65 subunit and p53 were decreased. EX527 (a specific SIRT1 inhibitor) conferred protection to the HUVECs against H2O2, as indicated by an improved cell viability, adhesion, an enhanced migratory ability, a decreased apoptotic index, decreased reactive oxygen species (ROS) production and reductions in several biochemical parameters. Immunofluorescence and Western blot analyses demonstrated that H2O2 treatment up-regulated SIRT1, phosphorylated-JNK (p-JNK), p-p38MAPK, and p-ERK expression. EX527 pretreatment reversed these effects on SIRT1, p-JNK, and p-p38MAPK but further increased the p-ERK levels. Similar results were confirmed in SIRT1 siRNA experiments. In summary, SIRT1 signaling pathway inhibition imparts protection against acute endothelial OSI, and modulation of MAPKs (JNK, p38MAPK, and ERK) may be involved in the protective effect of SIRT1 inhibition. © 2014 Elsevier B.V.
PubMed | Southern Medical University and The 97th Hospital of PLA
Type: | Journal: Photodiagnosis and photodynamic therapy | Year: 2016
To investigate the anti-tumor and immune efficacy of photodynamic immune-therapy (PIT), the combination of photodynamic therapy and dendritic cells (DC), on murine Heps hepatoma.DCs were derived from syngeneic mouse bone marrow and then labeled with DAPI in vitro. The hepatoma model was established by subcutaneous inoculation with Heps cells in one hundred and twenty-eight mice. They were then divided into four groups at random: control group, PDT group, DC group and PIT group. Tumors in the control group were injected with normal saline. Mice in the PDT group were injected with the photosensitizer Deuteporfin 24h before irradiation. Mice in the DC group were injected with DAPI labeled dendritic cells intratumorally. Mice in the PIT group were further given an injection of DCs after photoirradiation. Tumor growth and survival time were recorded after treatment. Fluorescence of tumor draining lymph nodes was evaluated under fluorescence microscope. Cytotoxic activity of splenocytes was tested by standard lactate dehydrogenase (lactate dehydrogenase, LDH) release assay.(1) Tumor growth was significantly slowed down in the PDT and PIT group compared to the control group (P<0.01). (2) The mean survival time was significantly prolonged in the PDT and PIT group. (3) The number of fluorescent cells in the draining lymph nodes from DC group was higher than that of the PIT group. (4) The anti-tumor activity of splenocytes in the PDT and PIT group was significantly higher than that of the DC and control groups (P<0.01, P<0.01).The present study suggests that PDT can inhibit tumor growth and induce anti-tumour immune response. The combination of PDT induced by Deuteporfiin and dendritic cell is capable of amplifying the antitumor immune response.
Liu Z.,The 309th Hospital of PLA |
Zhou G.,The 309th Hospital of PLA |
Deng X.,The 97th Hospital of PLA |
Yu Q.,The 309th Hospital of PLA |
And 6 more authors.
Journal of Infection | Year: 2014
Objectives: The regulatory mechanism of microRNA (miRNA) within macrophage innative response to Mycobacterium tuberculosis infection is not clear yet. Methods: The expression profile of cellular miRNAs during Mycobacterium bovis BCG infection was analyzed by using microarray. The expression of miR-146a was evaluated in alveolar macrophages (AMs) of bronchoalveolar lavage solution from pulmonary tuberculosis (PTB) patients and healthy volunteers respectively. Inhibitor experiment and promoter analysis were used to investigate the pathway involved in the induction of miR-146a. Examination of miR-146a function in macrophages was performed by overexpression and inhibition of miR-146a. Results: Among the altered miRNAs, 10 were downregulated whereas 8 were upregulated in M. bovis BCG-infected macrophage. MiR-146a was high expressed in cultured macrophage respond to M. bovis BCG but decreased in AMs of PTB patients, and stated a negative correlation with degree of smear-positive. Nuclear factor-κB pathway was required for the induction of miR-146a. Overexpression of miR-146a results in significant reduction of PTGS2 and enhanced the killing ability of THP-1 cells to intracellular M. bovis BCG, and miR-146a negatively regulated TNF-α release in feedback manner. Conclusions: Our findings suggest an important role of miR-146a in M. bovis BCG infection that helps to fine-tune the inflammation response of MTB infection. © 2014 The British Infection Association.
Shao G.-Q.,The 97th Hospital of PLA |
Zhang N.-Z.,The 97th Hospital of PLA
World Chinese Journal of Digestology | Year: 2014
AIM: To observe the effect of somatostatin on the expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in subcutaneous xenografts derived from human gastric carcinoma SGC-7901 cells in nude mice. METHODS: An animal model of human gastric cancer xenograft was established by subcutaneously implanting SGC-7901 cells in nude mice. Twenty-four nude mice were randomly divided into four groups and treated with normal saline (group A), octreotide (group B), high-dose somatostatin (group C) and low-dose somatostatin (group D) for three weeks, respectively. After treatment, the animals were killed to take the tumors. The expression of VEGF and bFGF was examined by immunohistochemistry and Western blot. RESULTS: Immunohistochemical analysis indicated that the integral optical density (IOD)/ area (/pix2) of VEGF in groups A-D were 0.644 ± 0.022, 0.549 ± 0.002, 0.345 ± 0.019 and 0.435 ± 0.018, respectively. The expression of VEGF protein in tumor tissue was significantly higher in group A than in the other three groups (P < 0.001). Compared with group B, the expression of VEGF was reduced more significantly in groups C and D (P < 0.001 for both). The IOD/ area (/pix2) of bFGF in groups A-D were 0.723 ± 0.018, 0.558 ± 0.004, 0.288 ± 0.017 and 0.595 ± 0.011, respectively. The expression of bFGF protein in tumor tissue was significantly higher in group A than in the other three groups (P < 0.001). Compared with group B, the expression of bFGF was reduced more significantly only in group C (P < 0.001). Western blot analysis indicated that the relative expression of VEGF in groups A-D were 0.98 ± 0.02, 0.76 ± 0.02, 0.53 ± 0.01 and 0.53 ± 0.01, respectively. The relative expression of bFGF in groups A-D were 0.76 ± 0.02, 0.71 ± 0.02, 0.32 ± 0.01 and 0.51 ± 0.01, respectively. The expression of bFGF and VEGF protein was significantly higher in group A than in the other three groups (P < 0.001). Compared with group B, the expression of VEGF and bFGF was reduced more significantly in groups C and D (P < 0.001). CONCLUSION: Somatostatin can down-regulate the expression of VEGF and bFGF in subcutaneous xenografts derived from human gastric carcinoma SGC-7901 cells in nude mice, and the effect is more obvious than that of octreotide. © 2014 Baishideng Publishing Group Inc. All rights reserved.
Luo X.M.,The 97th Hospital of PLA
Zhonghua nan ke xue = National journal of andrology | Year: 2012
To detect and compare the transcriptional activities of prostate-specific membrane antigen (PSMA) promoter and enhancer and survivin promoter in different human prostate cancer cell lines, and to search for some evidence for the targeting gene therapy of human prostate cancer. The fragments of the PSMA promoter and enhancer and survivin promoter were amplified by PCR and inserted into pGL3-Basic. The recombinant plasmids were transiently transfected into human prostate cancer cell lines and normal Chang liver cells, and, their transcriptional activities in various cells were determined by measuring the expression of luciferase. The survivin promoter exhibited a higher transcriptional activity than PSMA promoter and enhancer in tumor cell lines, and the S2pro promoter showed the highest activity, reaching one third of that of the CMV promoter. The survivin promoter is highly activated in prostate cancer cell lines and may serve as a new tool for the transcriptional targeting gene therapy of prostate cancer.
Dai X.-J.,Xuzhou Medical College |
Li N.,Xuzhou Medical College |
Yu L.,Xuzhou Medical College |
Chen Z.-Y.,Xuzhou Medical College |
And 3 more authors.
Cell Stress and Chaperones | Year: 2015
Microglia play an important role in neuronal protection and damage. However, the molecular and cellular relationship between microglia and neurons is unclear. We carried out a prospective study to detect that activation of BV2 microglia induced PC12 cell apoptosis in vitro through the TLR4/adapter protein myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) signaling pathway. BV2 microglia were treated with different concentrations of LPS for 24 h. Western blot was utilized to detect the expression of TLR4 and the downstream signaling pathway. The level of inflammatory mediator was quantified using a specific ELISA kit. The supernatant of 10 μg/ml LPS-treated BV2 cells was used as conditioned medium (CM). PC12 cells were co-culture with CM for 24 h. Cell viability was determined by MTT assay and cell apoptosis was tested by flow cytometry. BV2 microglia were treated with 10, 20, or 30 μg/ml LPS for 24 h. The expression of TLR4, MyD88, and NF-κB significantly increased. When PC12 cells were co-cultured with CM for 24 h, cell viability decreased. CM up-regulated the Bax level and down-regulated the Bcl-2 protein level in PC12 cells. PC12 cells pretreated with interleukin-1 receptor antagonist (IL-1RA) for 30 min, significantly alleviated CM-induced PC12 cell apoptosis. These results suggest that BV2 microglia activated by LPS triggered TLR4/MyD88/NF-κB signaling pathway that induced the release of IL-1β and could participate in the PC12 cells injury. © 2014, Cell Stress Society International.
PubMed | the 97th Hospital of PLA
Type: Journal Article | Journal: Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns | Year: 2013
To observe the effects of glutamine combined with ulinastatin on inflammatory response of patients with severe burn injury.Sixty patients with severe burn injury admitted to our burn wards from January 2010 to December 2011 conforming to the study criteria were divided into control group (C, n = 20), glutamine group (G, n = 20), and glutamine combined with ulinastatin group (G + U, n = 20) according to the random number table. Another 10 healthy volunteers were chosen as normal control group (NC). Isonitrogenous and isocaloric nutrition supports were given to patients in groups C, G, and G + U from post burn day (PBD) 2. 0.3 g/kg protein in the form of glutamine dipeptide was given to patients in group G for 10 days. 0.3 g/kg protein was given to patients in group G + U for 10 days with the same amount of glutamine dipeptide as that in group G, followed by intravenous injection of 100 kU ulinastatin (once per 8 hours) for 7 days during 10 days. The nitrogen concentration of 24 h urine was determined with Kieldahl nitrogen determination method, and nitrogen balance was calculated one day before treatment and ten days after treatment. Meanwhile, the levels of D-lactate in serum was determined by colorimetric method, the levels of diamine oxidase (DAO), TNF-, and IL-6 by enzyme-linked immunosorbent assay, and LPS level by kinetic turbidimetric assay with TAL. Above-mentioned indexes were also examined in group NC. The wound healing rate on PBD 30, total hospital stay days, and the incidence of burn sepsis of all burn patients were recorded. Data were processed with one-way analysis of variance, LSD test, t test, and chi-square test.Compared with that in group C [(-5.40 1.67) g/d], nitrogen balance in group G was significantly increased ten days after treatment [(-1.35 0.59) g/d, P < 0.01]. The serum levels of D-lactate, DAO, LPS, TNF-, and IL-6 in group G ten days after treatment were significantly lower than those in group C (P < 0.05 or P < 0.01). No statistically significant difference was observed in nitrogen balance and the serum levels of D-lactate, DAO between group G + U and group G (P values all above 0.05). The serum levels of LPS, TNF-, and IL-6 in group G + U ten days after treatment were respectively (0.167 0.064) EU/mL, (43 14) pg/mL, (139 23) pg/mL, which were significantly lower than those in group G [(0.240 0.079) EU/mL, (59 8) pg/mL, (195 31) pg/mL, respectively, P < 0.05 or P < 0.01]. The would healing rate on PBD 30 and total hospital stay days in group G were respectively higher and shorter than those in group C (P values all below 0.01), but no statistically significant difference in the incidence of burn sepsis was found between them (P > 0.05). The would healing rate on PBD 30 in group G+U [(96 4)%] was enhanced, and total hospital stay days [(41 4) d] were lowered than those in group G [(88 7)%, (49 5)d, P values all below 0.01]. The incidence of burn sepsis of patients in group G + U (5%) was significantly lower than that in group C (35%, (2) = 6.234, P < 0.05).Glutamine combined with ulinastatin treatment can alleviate damage to intestine after severe burn injury, lower the serum level of inflammatory cytokines, promote wound healing, and reduce the incidence of burn sepsis.
PubMed | Xuzhou Medical College and The 97th Hospital of PLA
Type: Journal Article | Journal: Injury | Year: 2014
Posttraumatic immune disorder can cause complications including systemic inflammatory response syndrome (SIRS) and multiple-organ dysfunction syndrome (MODS). Cytotoxic granules containing perforin and granzyme-B (GrB) are released by cytotoxic CD8(+) T lymphocytes, NK and T cells after major trauma. This prospective clinical study was designed to analyze the association between these immune components and complications after major trauma.We retrospectively studied 48 patients aged between 16 and 65 years admitted within 90min of major trauma (Injury Severity Score>16) and surviving beyond 7 days, and 20 healthy controls. Blood samples were drawn on admission and after 1, 3 and 7 days. CD8(+) T, NK and T cell counts in peripheral blood and the levels of perforin and GrB in these cells were analyzed by flow cytometry. Clinical aspects of MODS and SIRS were recorded daily.CD8(+) T cell counts were not significantly different in patients with SIRS or uncomplicated group, but were depressed in the MODS group after trauma. However, NK cell counts in patients with MODS were significantly depressed only at day 7 after injury, and T cell counts were significantly depressed after trauma. Perforin levels in CD8(+) T, NK and T cells in patients with MODS were depressed after trauma. GrB levels in NK, CD8(+) T and T cells in patients with MODS were significantly depressed at 3 and 7 days post trauma.Posttraumatic MODS is associated with early, sustained, and severe depression of lymphocytes.