RBM4 modulates the proliferation and expression of inflammatory factors via the alternative splicing of regulatory factors in HeLa cells.

Regulatory factors function by modulating a variety of cascade mechanisms in cells. RBM4 is a multifunctional RNA-binding protein in post-transcriptional gene regulation. Cytoplasmic RBM4 interacts with Ago2 to regulate inflammatory responses by affecting mRNA decay and cap-dependent translation. However, it is unclear whether RBM4 functions in inflammation regulation by its splicing factor role. Here, the cell biology, gene expression profile and alternative splicing pattern of HeLa cells with RBM4 overexpression (RBM-OE) were compared with the control. The results showed that RBM4-OE inhibited proliferation. RBM4-OE extensively affects the transcriptional level of genes involved in cell surface receptor signalling pathway, inflammatory responses and the response to lipopolysaccharide. RBM4 broadly regulated the alternative splicing of hundreds of genes with functions of protein binding, helicase activity, DNA binding and transcription co-activator. RBM4-regulated splicing of these genes plays an important role in apoptotic process and gene transcription regulation. As an example, exon inclusion of TNIP1 mediated by RBM4 affects the expression of its targets in inflammatory pathways. These results indicated that RBM4 can mediate the inflammatory response via splicing regulation, which adds to the understanding of the critical role of RBM4 in cancer complicated by inflammation. In conclusion, this study indicated a mechanism in which the dysregulation of alternative splicing can influence cellular biology and lead to various immune-related diseases.

Caspase-1 inhibitor exerts brain-protective effects against sepsis-associated encephalopathy and cognitive impairments in a mouse model of sepsis.

Sepsis-associated encephalopathy (SAE) manifested clinically in acute and long-term cognitive impairments and associated with increased morbidity and mortality worldwide. The potential pathological changes of SAE are complex and remain to be elucidated. Pyroptosis, a novel programmed cell death, is executed by caspase-1-cleaved GSDMD N-terminal (GSDMD-NT) and we investigated it in peripheral blood immunocytes of septic patients previously. Here, a caspase-1 inhibitor VX765 was treated with CLP-induced septic mice. Novel object recognition test indicated that VX765 treatment reversed cognitive dysfunction in septic mice. Elevated plus maze, tail suspension test and open field test revealed that depressive-like behaviors of septic mice were relieved. Inhibited caspase-1 suppressed the expressions of GSDMD and its cleavage form GSDMD-NT, and reduced pyroptosis in brain at day 1 and day 7 after sepsis. Meantime, inhibited caspase-1 mitigated the expressions of IL-1ß, MCP-1 and TNF-α in serum and brain, diminished microglia activation in septic mice, and reduced sepsis-induced brain-blood barrier disruption and ultrastructure damages in brain as well. Inhibited caspase-1 protected the synapse plasticity and preserved long-term potential, which may be the possible mechanism of cognitive functions protective effects of septic mice. In conclusion, caspase-1 inhibition exerts brain-protective effects against SAE and cognitive impairments in a mouse model of sepsis.

Chinese Trauma Surgeon Association for management guidelines of vacuum sealing drainage application in abdominal surgeries-Update and systematic review.

Vacuum sealing drainage (VSD) is frequently used in abdominal surgeries. However, relevant guidelines are rare. Chinese Trauma Surgeon Association organized a committee composed of 28 experts across China in July 2017, aiming to provide an evidence-based recommendation for the application of VSD in abdominal surgeries. Eleven questions regarding the use of VSD in abdominal surgeries were addressed: (1) which type of materials should be respectively chosen for the intraperitoneal cavity, retroperitoneal cavity and superficial incisions? (2) Can VSD be preventively used for a high-risk abdominal incision with primary suture? (3) Can VSD be used in severely contaminated/infected abdominal surgical sites? (4) Can VSD be used for temporary abdominal cavity closure under some special conditions such as severe abdominal trauma, infection, liver transplantation and intra-abdominal volume increment in abdominal compartment syndrome? (5) Can VSD be used in abdominal organ inflammation, injury, or postoperative drainage? (6) Can VSD be used in the treatment of intestinal fistula and pancreatic fistula? (7) Can VSD be used in the treatment of intra-abdominal and extra-peritoneal abscess? (8) Can VSD be used in the treatment of abdominal wall wounds, wound cavity, and defects? (9) Does VSD increase the risk of bleeding? (10) Does VSD increase the risk of intestinal wall injury? (11) Does VSD increase the risk of peritoneal adhesion? Focusing on these questions, evidence-based recommendations were given accordingly. VSD was strongly recommended regarding the questions 2-4. Weak recommendations were made regarding questions 1 and 5-11. Proper use of VSD in abdominal surgeries can lower the risk of infection in abdominal incisions with primary suture, treat severely contaminated/infected surgical sites and facilitate temporary abdominal cavity closure.

Cyclooxygenase-2 blockade inhibits accumulation and function of myeloid-derived suppressor cells and restores T cell response after traumatic stress.

Myeloid-derived suppressor cells (MDSCs) play a crucial role in T cell dysfunction, which is related to poor outcome in patients with severe trauma. Cyclooxygenase-2 (Cox-2) contributes to immune disorder in trauma and infection via production of prostaglandin E2. However, the role of Cox-2 in the accumulation and function of MDSCs after traumatic stress has not been fully elucidated. In the present study, we treated murine trauma model with NS398, a selective Cox-2 inhibitor. Then the percentages of CD11b+/Gr-1+ cells, proliferation and apoptosis of CD4+ T cells were determined. Arginase activity and arginase-1 (Arg-1) protein expression of splenic CD11b+/Gr-1+ cells, and delayed-type hypersensitivity (DTH) response were analyzed. The results showed that Cox-2 blockade significantly decreased the percentages of CD11b+/Gr-1+ cells in the spleen and bone marrow 48 and 72 h after traumatic stress. NS398 inhibited arginase activity and down-regulated the Arg-1 expression of splenic CD11b+/Gr-1+ cells. Moreover, NS398 could promote proliferation and inhibit apoptosis of CD4+ T cells. It also restored DTH response of traumatic mice. Taken together, our data revealed that Cox-2 might play a pivotal role in the accumulation and function of MDSC after traumatic stress.

Inhibition of the C3a receptor attenuates sepsis-induced acute lung injury by suppressing pyroptosis of the pulmonary vascular endothelial cells.

Acute lung injury (ALI) is the leading cause of bacterial sepsis-related death because of disrupted pulmonary endothelial barrier, resulting in protein-rich pulmonary oedema, an influx of pro-inflammatory cells and refractory hypoxaemia. Several studies have reported that C3a levels are significantly higher in organs with sepsis and their peripheral organs and are closely associated with organ dysfunction and poor prognosis in sepsis. However, the role of the C3a complement in sepsis ALI remains unclear. Therefore, this study aimed to investigate the important role and mechanism of C3a in preventing the occurrence of pyroptosis (a pro-inflammatory form of cell death) to protect the lung endothelial cells (ECs) in sepsis-induced ALI. A septic mouse model was established with cecal ligation and puncture (CLP), which demonstrated that C3a mediated EC pyroptosis through its C3aR receptor. Furthermore, inhibition of the C3a-C3aR axis could block both NLRP3/caspase-1 and caspase-11 pathways, thus preventing pulmonary EC from pyroptosis. These results indicate that inhibition of the C3A-C3AR complement axis can inhibit pulmonary vascular EC pyroptosis, a potential target for the treatment of ALI.

Analysis of misdiagnosis in patients with multiple trauma.

OBJECTIVE: To explore the features, treatment outcomes and reasons for misdiagnosis in patients with multiple trauma, so as to decrease the incidence of misdiagnosis. METHODS: A total of 3 163 patients with multiple trauma who were admitted in our department from August 1997 to August 2008, were retrospectively studied to compare the features of diagnosis and treatment. There were 2 117 males (66.93%) and 1 046 females (33.07%) with the mean age of 36.46 years (range, 14-80 years). Parameters such as general status, traumatic condition, diagnosis and treatment situation, prognosis and mortality were analyzed. The differences between misdiagnosis group and correct diagnosis group were compared in terms of severity of injury, complications and treatment outcomes to elucidate the cause and prevention of misdiagnosis. RESULTS: The misdiagnosis rate of multiple trauma in this study was 16.19%. The major anatomic sites misdiagnosed were limbs and pelvis (299 positions, 39.50%), abdominal region and pelvic organ (148 positions, 19.55%), and thoracic region (109 positions, 14.40%). In misdiagnosis group, ISS, length of hospital stay, rates of disturbance of consciousness, critical cases and shock cases were 33.78+/-19.64, (23.59+/-7.26) days, 49.22%, 33.01% and 47.46%, respectively, which were significantly higher than those of the correct diagnosis group (P less than 0.01). And the data showed that the more serious the injury was, the higher the rate of misdiagnosis would be. The rate of primary diagnosis by trauma surgeons in correct diagnosis group was 75.78%, significantly higher than that of the misdiagnosis group (X(2) equal to 382.01, P less than 0.01). The mortality rate of the misdiagnosis group was 2.93%, which was significantly higher than that for all patients (X(2) equal to 5.22, P less than 0.05). CONCLUSIONS: The results indicated that patients with severe multiple trauma are at high risk of misdiagnosis in early treatment. The mortality rate of misdiagnosed patients is higher than the correctly-diagnosed patients. To prevent misdiagnosis, physicians need to take great care to conduct thorough clinical examinations and repeated evaluation.

Modified lumbopelvic fixation for sacral and L5 fractures associated with spinopelvic instability: a case report and introduction of the surgical technique.

Posterior lumbopelvic fixation with iliac screws is the most commonly used method for unstable spinopelvic injuries. It has certain limitations including inability to use distraction along the spinopelvic rod as an indirect reduction maneuver, need for complex 3-dimensional rod contouring and complications such as hardware prominence and soft tissue coverage. In the present case report, we described a surgical technique of lumbopelvic fixation with sacral alar screws for traumatic spinopelvic instability resulted from a unilateral Denis-III comminuted sacral fracture and the L5 burst fracture. On the opposite side of the sacral fracture, caudal screws were implanted into the pedicle of the S1, whereas on the side of sacral fracture, two sacral alar screws were placed parallel to the superior sacral endplate as well as the plane of sacroiliac joint. In addition, horizontal stabilization was conducted with cross-link connections to maintain the longitudinal traction. For sacral fracture associated with traumatic spinopelvic instability, this modified lumbopelvic fixation technique using sacral alar screws makes longitudinal reduction easier, requires less rod contouring, and reduces hardware prominence without compromising the stability.

Morin exerts protective effects on encephalopathy and sepsis-associated cognitive functions in a murine sepsis model.

Sepsis-associated encephalopathy (SAE) often leads to cognitive impairments in the rest life of septic survivors. The potential pathological changes of SAE are complicated and have not been fully understood. Morin, a flavone compound exhibiting neuroprotective activity and anti-inflammation effect, was employed to treat with CLP-induced septic mice in our study. The data from a novel object recognition test and tail suspension test indicated that morin treatment reversed cognitive dysfunction and relieved depressive-like behaviors in septic mice. Morin down-regulated the expressions of IL-6, MCP-1, TNF-α and IL-10 in serum and diminished microglia activation in septic mice. Additionally, Western blot results showed that morin reduced the phosphokinase GSK3ß activity and elevated the phosphatase PP2A activity, which led to lower tau phosphorylation. Morin reduced Aß deposition and protected the synapse integrity, which might be the possible mechanism of protecting cognitive functions in septic mice. In conclusion, we identified that morin exerted anti-inflammation and anti-neurodegeneration effects in septic mice, and prevented further cognitive impairments.

FK866 attenuates sepsis-induced acute lung injury through c-jun-N-terminal kinase (JNK)-dependent autophagy.

AIMS: Increasing evidence indicates that FK866, a specific noncompetitive nicotinamide phosphoribosyl transferase inhibitor, exhibits a protective effect on acute lung injury (ALI). Autophagy plays a pivotal role in sepsis-induced ALI. However, the contribution of autophagy and the underlying mechanism by which FK866-confered lung protection remains elusive. Herein, we aimed to study whether FK866 could alleviate sepsis-induced ALI via the JNK-dependent autophagy. MAIN METHODS: Male C57BL/6 mice were subjected to cecal ligation and puncture (CLP) to establish the polymicrobial sepsis mice model, and treated with FK866 (10 mg/kg) at 24, 12 and 0.5 h before the CLP procedure. The lung protective effects were measured by lung histopathology, tissue edema, vascular leakage, inflammation infiltration, autophagy-related protein expression and JNK activity. A549 cells were stimulated with LPS (1000 ng/ml) to generate the ALI cell model, and pretreated with FK866 or SP600125 for 30 min to measure the autophagy-related protein expression and JNK activity. KEY FINDINGS: Our results demonstrated that FK866 reduced lung injury score, tissue edema, vascular leakage, and inflammatory infiltration, and upregulated autophagy. The protective effect of autophagy conferred by FK866 on ALI was further clarified by using 3-methyladenine (3MA) and rapamycin. Additionally, the activity of JNK was suppressed by FK866, and inhibition of JNK promoted autophagy and showed a benefit effect. SIGNIFICANCE: Our study indicates that FK866 protects against sepsis-induced ALI by induction of JNK-dependent autophagy. This may provide new insights into the functional mechanism of NAMPT inhibition in sepsis-induced ALI.

[Expression of soluble human triggering receptors on myeloid cell-1 in patients with multiple trauma and its clinical significance].

OBJECTIVE: To investigate the expression profile of human soluble triggering receptor on myeloid cell-1 (sTREM-1) in patients with multiple trauma and determine its clinical significance. METHODS: Peripheral blood of 52 patients admitted to the hospital from October 2007 to January 2008 with multiple traumas with injury severity score (ISS) > or = 16 and 7 healthy volunteers were obtained, and sera samples were isolated. sTREM-1 was determined by semi-quantitative immunoblot technique. TNF-alpha and C-reactive protein (CRP) were determined by ELISA. RESULTS: sTREM-1 of patients with multiple traumas was significantly increased as compared with that of control (P < 0.001), and sTREM-1 of ISS > or = 25 group was significantly higher than that of 16 < or = ISS < 25 group (P < 0.05). sTREM-1 level correlated closely with TNF-alpha level (r = 0.845, P < 0.05), but did not correlate with CRP (r = 0.426, P > 0.05). In patients with sepsis, sTREM-1 on 1, 2 and 7 d was (25.1 +/- 2.2), (31.9 +/- 2.6) and (25.2 +/- 1.9) ng/L, respectively. In patients without sepsis, sTREM-1 on 1, 2 and 7 d was (15.8 +/- 1.3), (24.2 +/- 2.0) and (13.9 +/- 1.5) ng/L, respectively. sTREM-1 of patients with sepsis was significantly higher than that of patients without sepsis (P < 0.05). CONCLUSIONS: Serum sTREM-1 correlates closely with ISS, TNF-alpha and onset of sepsis, indicating that it may play an important role in the development of sepsis in patients with multiple traumas.

Dihydromyricetin Alleviates Sepsis-Induced Acute Lung Injury through Inhibiting NLRP3 Inflammasome-Dependent Pyroptosis in Mice Model.

Increasing evidence demonstrates that pyroptosis, pro-inflammatory programmed cell death, is linked to acute lung injury (ALI). Dihydromyricetin (DHM) has been reported to exert anti-inflammatory effects by inhibiting NLRP3 inflammasome activation in vascular endothelial cells. However, the effects of DHM on NLRP3 inflammasome-induced pyroptosis in ALI remain elusive. In the present study, male BALB/c mice were subjected to cecal ligation and puncture (CLP), and DHM (50, 100, 150 mg/kg) was orally administered (once per day, for 3 days) 2 h after CLP. After 72 h, lung histopathology was examined, and the wet/dry (W/D) ratio, inflammatory infiltration, total protein concentration, total cell, and neutrophil counts were detected. Myeloperoxidase (MPO), interleukin (IL)-6, TNF-α, IL-1ß, and IL-18 levels in bronchoalveolar lavage fluid (BALF) were measured by ELISA. Additionally, the expression of NLRP3 signaling pathway proteins were detected by Western blotting. The results revealed that in BALF, DHM (150 mg/kg) treatment significantly reduced the CLP-induced lung histopathological injury, inflammatory cell infiltration, total cell and neutrophil number, and total protein and albumin concentration. DHM treatment significantly inhibited the CLP-induced NLRP3 inflammasome pathway (NLRP3, ASC, caspase-1, gasdermin D (Gsdmd), IL-1ß, and IL-18). In conclusion, these results demonstrate that DHM protects against CLP-induced ALI by inhibiting NLRP3 inflammasome activation and subsequent pyroptosis.

MicroRNA Dysregulation and Steroid Hormone Receptor Expression in Uterine Tissues of Rats with Endometriosis during the Implantation Window.

BACKGROUND: Estrogen receptor (ER) and progesterone receptor (PR) are involved in endometriosis, but the involvement of microRNAs (miRNAs) is unknown. The aim of the study was to explore the correlation between miRNA and ER/PR in uterine tissues of rats with endometriosis during the implantation window. METHODS: Twenty female Sprague-Dawley rats were randomized in three groups: endometriosis (n = 7), fat tissue control (n = 6), and normal (n = 7) groups. The female rats were mated and sacrificed on day 5 (implantation). Uterine tissues were obtained for hematoxylin-eosin staining, immunohistochemistry, and miRNA expression. Reverse transcription polymerase chain reaction (RT-PCR) was used to validate the expression of rno-miR-29c-3p, rno-miR-34c-5p, rno-miR-141-5p, rno-miR-24-1-5p, and rno-miR-490-5p. RESULTS: The 475 miRNAs were found to differentially express between the endometriosis and normal control groups, with 127 being upregulated and 348 being downregulated. Expression of five miRNAs (rno-miR-29c-3p, rno-miR-34c-5p, rno-miR-141-5p, rno-miR-24-1-5p, and rno-miR-490-5p) were validated by RT-PCR and found to be differentially expressed among the three groups. Expression of ER and PR proteins (immunohistochemistry) in the glandular epithelium and endometrial stroma was significantly different among the three groups (all P < 0.05). Five miRNAs were involved in pathways probably taking part in implantation and fertility. CONCLUSIONS: The results suggested that miRNAs, ER, and PR could play important roles in the embryo implantation period of rats with endometriosis. These miRNAs might play a role in endometrial receptivity in endometriosis.

Caspase-1-dependent pyroptosis of peripheral blood mononuclear cells predicts the development of sepsis in severe trauma patients: A prospective observational study.

Pyroptosis plays a pivotal role in sepsis and septic shock in animal studies. However, its clinical significance in pathological conditions has not been well elucidated. This study aimed to evaluate the correlation between the percentage of pyroptotic peripheral blood mononuclear cells (PBMCs) and the clinical index and to investigate the relationship between PBMCs pyroptosis and the development of sepsis in trauma patients.This prospective study was conducted from October 2016 to May 2017 in a comprehensive trauma center. Sixty trauma patients and 10 healthy controls were enrolled. Peripheral blood samples were collected from the patients within 24 hours after injury. The percentages of pyroptotic and apoptotic PBMCs were measured using flow cytometry, and plasma levels of cytokines were evaluated using flow cytometric analysis with a human inflammation 13-plex panel.Trauma patients who developed sepsis had higher percentages of pyroptotic and apoptotic PBMCs at admission. Patients who developed sepsis (n = 33) had higher interleukin (IL)-6, IL-18, and monocyte chemotactic protein-1 (MCP-1) concentrations at admission than patients (n = 27) who did not develop sepsis. The percentage of PBMCs pyroptosis was significantly correlated with injury severity score (ISS), acute physiology and chronic health evaluation (APACHE) II score, IL-10, IL-18, and MCP-1 levels in trauma patients. PBMCs pyroptosis is a better biomarker in predicting the development of sepsis after trauma.This study indicates that the percentage of pyroptotic PBMCs increases during the early phase of trauma and that this increase is significantly correlated with the severity and state of inflammation in trauma patients. PBMCs pyroptosis is a potential marker for predicting the development of sepsis after trauma.

[Analysis of emergency treatment in 4519 patients with multiple injuries].

OBJECTIVE: To investigate the clinical manifestations, treatment, complications, and prognosis of patients with multiple injuries. METHOD: The clinical data, including the causes of injury, treatment, complications, causes of death, and mortality rate, of 4519 patients were retrospectively analyzed. RESULTS: The major causes of injury were road traffic injury (2410 cases, 53.33%), violence injury (747 cases, 16.53%), and high falling injury (575 cases, 12.72%). The main involved positions included head (2247 cases, 18.71%), abdominal region and pelvis (2118 cases, 17.64%), and thoracic region (1853 cases, 15.43%). The major complications were shock (1497 cases, 33.13%). The main cause of death was sepsis with multiple organ dysfunction syndrome/failure (28 cases, 82.35%) after multiple injuries, significant higher than other causes in the same period (P<0.01). CONCLUSIONS: The multiple injuries have various causes of disease, and were complicated with their diverse clinical manifestations, numerous complications, and high mortalities. Further research on the integrated rescue mortality is required.

Predictive factors of ovarian carcinoma for women with ovarian endometrioma aged 45 years and older in China.

BACKGROUND: To explore the risk factors of endometriosis-associated ovarian cancer (EAOC) in women with ovarian endometriosis (OEM) aged 45 years and above in China. METHODS: This study reviewed the medical records of 1038 women in total who were aged 45 years and above, surgical-pathologically diagnosed with ovarian endometriosis, and were treated at Peking Union Medical College Hospital between December 1996 and December 2016. Histology evaluation was used to determine whether the ovarian endometriosis specimen was with (n = 30) or without (n = 1008) ovarian cancer. RESULTS: 2.9% (30/1038) of women with the surgical-pathological diagnosis of ovarian endometriosis were found to have EAOC. Those patients with EAOC were prone to be in the postmenopausal status at the time of the diagnosis (OR 5.50, 95%CI 2.54-11.90, P < .001) and larger size of tumor (≥8 cm, OR 7.19, 95% CI 3.34-15.50, P < .001), and higher prevalence of coexisting with endometrial disorders (OR 4.11, 95%CI 1.73-9.73, P = .003). This study showed that patients of an older age when diagnosed with OEM, were at a higher risk of developing EAOC, respectively measuring of 1.7% (13/751) at 45-49 years, 5.6% (12/215) at 50-54 years, and 10.0%(5/50) at 55-59 years (P < 0.001). CONCLUSIONS: This study showed that for women aged 45 years and above who were diagnosed with OEM, the independent risk factors of EAOC were menopausal status, tumor size of 8 cm or greater in diameter, and coexisting endometrial disorders. Therefore, intensive follow-ups or active interventions should be considered for them.

The influence of sex on outcomes in trauma patients: a meta-analysis.

BACKGROUND: This study aims to assess the influence of sex on outcomes among trauma patients, including injury severity, medical resource utility, complications, and mortality. METHODS: A systematic review of the literature was conducted by internet search. Data were extracted from selected studies and analyzed using Stata to compare outcomes between male and female injured patients. RESULTS: Eventually, 19 studies met our inclusion criteria with 100,566 men and 39,762 women. Pooled data revealed that male sex was associated with increased risk of mortality, hospital length of stay, and higher incidence of complications. No difference was detected in injury severity between male and female patients. CONCLUSION: Evidence of this meta-analysis strongly supports the sex dimorphism in the prognosis of trauma patients and further work should be done to decipher potential mechanism.

Hepatitis B virus X gene induces human telomerase reverse transcriptase mRNA expression in cultured normal human cholangiocytes.

AIM: To study the transcriptional regulation of human telomerase reverse transcriptase (hTERT) mRNA in normal human cholangiocytes (HBECs) after hepatitis B virus X (HBx) gene transfection and to elucidate the possible mechanism of HBV infection underlying cholangiocarcinoma. METHODS: HBECs were cultured in vitro and co-transfected with a eukaryotic expression vector containing the HBx coding region and a cloning vector containing coding sequences of enhanced green fluorescent protein (EGFP) using lipid-mediated gene transfer. The transfection efficiency was determined by the expression of EGFP. The expressions of hTERT mRNA and HBx protein in HBECs were detected by RT-PCR and immunocytochemical stain, respectively. RESULTS: The transfection efficiencies were about 15% for both HBx gene expression plasmid and empty vector. No hTERT mRNA was expressed in HBECs when transfected with OPTI-MEM medium and empty vector, but a dramatic increase was observed for hTERT mRNA expression in HBECs when transfected with HBx expression vector. HBx protein was only expressed in HBECs when transfected with HBx expression vector. CONCLUSION: HBx transfection can activate the transcriptional expression of hTERT mRNA. Cis-activation of hTERT mRNA by HBx gene is the primary mechanism underlying the proliferation, differentiation and tumorigenesis of biliary epithelia.

Overexpression of mitofusin 2 inhibits reactive astrogliosis proliferation in vitro.

Astrocytes become activated in response to central nervous system (CNS) injury, and excessive astrogliosis is considered an impediment to axonal regeneration by forming glial scar. Mitofusin 2 (Mfn2), a key protein in mitochondrial network, has been reported to negatively regulate cell proliferation. The present study aimed to explore whether reactive astrogliosis could be suppressed by Mfn2 overexpression. Scratch injury and starvation-serum stimulation models in cultured astrocytes were combined to address this issue. In scratch model, reactive proliferation status of damaged astrocytes was implicated by migration of high ratio of EdU(+) cells into lesion region and significantly increased expression of GFAP and PCNA. At meantime, Mfn2 expression was found to exert a down-regulated trend both in gen and protein levels. Pretreatment of cells with adenoviral vector encoding Mfn2 gene increased Mfn2 expression and subsequently attenuated injury-induced astrocytes hyperplasia, activation-relevant protein synthesis, cellular proliferation, eventually delayed wound healing process. Furthermore, Mfn2 overexpression markedly inhibited astrocytes proliferation induced by serum stimulation, by arresting the transition of cell cycle from G1 to S phase. Together, these in vitro results demonstrated that reactive astrogliosis can be effectively suppressed by up-regulation of Mfn2, which might contribute to a promising therapeutic intervention in CNS disease characterized by glia-related damage.

Effect of triptolide on secretion of inflammatory cellular factors TNF-α and IL-8 in peritoneal macrophages of mice activated by lipopolysaccharide.

BACKGROUND: Research has been carried out to look for safe and effective anti-inflammation drugs from traditional Chinese herbal medicine. As a powerful research technology of life science, molecular biology has entered many areas of traditional Chinese medicine. This study aimed to investigate the effect of triptolide on tumor necrosis factor-a (TNF-α) and interleukin-8 (IL-8) of peritoneal macrophages activated by lipopolysaccharide (LPS) in mice. METHODS: Peritoneal elicited macrophages were separated, purified and activated by LPS in mice, then cultured in vitro with triptolide at different concentrations. The activity of TNF-α and the level of IL-8 of cellular supernatants were determined by MTT colorimetric assay and ELISA, respectively. RESULTS: The activity of TNF-α in macrophages was significantly inhibited (P<0.01) by triptolide (10(-1)-10(1)µg/ml) during 4-24 hours in a time- and dose-dependent manner. The level of IL-8 in macrophages was significantly inhibited (P<0.01) by triptolide (10(-1)-10(1)µg/ml) in 12 hours in a dose-dependent manner. CONCLUSION: Triptolide could inhibit the activity of TNF-α and the level of IL-8 in macrophages activated by LPS.

Analysis of olfactory ensheathing glia transplantation-induced repair of spinal cord injury by electrophysiological, behavioral, and histochemical methods in rats.

This study examined the efficacy of transplanting olfactory ensheathing glia (OEG) in repairing spinal cord injury (SCI) using behavioral tests, retrograde labeling, as well as somatosensory and motor evoked potentials in rats. One week after surgery, motor function in OEG-treated rats was significantly superior to untreated controls (P < 0.05). Also, we found that up to 8 weeks following surgery to induce SCI, somatosensory and motor evoked potentials were found in the OEG-treated groups, but not in the transplantation and damage control groups. Retrograde labeling from the area distal to the SCI produced a higher number of labeled neurons in the ventrolateral division of red nucleus and motor cortex of OEG-treated rats compared to controls, which showed no retrograde labeling (P < 0.05). We believe that this study has important implications for characterizing the mechanisms of OEG transplantation as a treatment for SCI.