Dexmedetomidine mitigates lipopolysaccharide-induced acute lung injury by modulating heat shock protein A12B to inhibit the toll-like receptor 4/nuclear factor-kappa B signaling pathway.

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS): Life-threatening medical conditions characterized by high morbidity and mortality rates, where the inflammatory process plays a crucial role in lung tissue damage, especially in models induced by lipopolysaccharide (LPS). Heat shock protein A12B (HSPA12B) has strong anti-infammatory properties However, it is unknown whether increased HSPA12B is protective against LPS-induced ALI. And Dexmedetomidine (DEX) is a potent α2-adrenergic receptor (α2-AR) agonist that has been shown to protect against sepsis-induced lung injury, however, the underlying mechanisms of this protection are not fully understood. This study utilized bioinformatics analysis and an LPS-induced ALI model to explore how DEX alleviates lung injury by modulating HSPA12B and inhibiting the Toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB) signaling pathway. Results indicate that HSPA12B overexpression and DEX pre-treatment markedly mitigated LPS-induced lung injury, which was evaluated by the deterioration of histopathology, histologic scores, the W/D weight ratio, and total protein expression, tumor necrosis factor-alpha (TNF-α), and interleukin-1ß (IL-1ß) in the BALF, and the levels of NO, MDA,SOD and MPO in the lung. Moreover, HSPA12B overexpression and DEX pre-treatment significantly reduces lung injury and inflammation levels by upregulating HSPA12B and inhibiting the activation of the TLR4/NF-κB signaling pathway. On the contrary, when the expression of HSPA12B is inhibited, the protective effect of DEX pre-treatment on lung tissue is significantly weakened.In summary, our research demonstrated that the increased expression of AAV-mediated HSPA12B in the lungs of mice inhibits acute inflammation and suppresses the activation of TLR4/NF-κB pathway in a murine model of LPS-induced ALI. DEX could enhance HSPA12B and inhibit the initiation and development of inflammation through down-regulating TLR4/NF-κB pathway.These findings highlight the potential of DEX as a therapeutic agent for treating ALI and ARDS, offering new strategies for clinical intervention.

Effect of Dexmedetomidine on Apoptosis and Early Cognitive Dysfunction in Rats Following Propofol Anesthesia.

Background: To address the intricate interplay between surgical anesthesia, hippocampal apoptosis, and early cognitive dysfunction in rats, this study delves into the impact of dexmedetomidine (DEX). With a focus on understanding the alterations in the PI3K/Akt pathway, our investigation aims to shed light on the comprehensive dynamics of these processes. Methods: Sixty healthy Sprague-Dawley rats were randomly divided into three groups: controls (intraperitoneal saline injection), group A (intraperitoneal propofol injection), and group A + DEX (intraperitoneal propofol and DEX injection), with 20 rats in each group. Cognitive function in the three groups was evaluated using [specify the cognitive function tests, e.g., Morris Water Maze]. The assessment was conducted at various postoperative time points. We employed Hematoxylin-eosin (HE) staining, flow cytometry (FC), Western blot (WB), and real-time fluorescence quantitation polymerase chain reaction (RT-qPCR) to examine hippocampal apoptosis and gene expression. Results: Rats in groups A and A + DEX exhibited higher cognitive scores (NSS) and escape latencies at each postoperative time point, along with a lower proportion of swimming distance (SD) in the target quadrant (TQ) compared to controls. Group A + DEX demonstrated lower NSS and escape latency than controls, accompanied by a higher proportion of SD in TQ. Apoptosis rates of hippocampal cells were higher in groups A and A + DEX than in controls, although they were relatively lower in group A + DEX (P < .05). Regarding gene expression, the relative expression (RE) of Bcl-2, PI3K, pAkt, mRNA expression of Bcl-2 and PI3K, and Bax and caspase3 were altered. Group A + DEX showed higher RE of Bcl-2, PI3K, pAkt, mRNA expression of Bcl-2 and PI3K, and lower Bax and caspase3 than group A (P < .05). For a detailed understanding of the magnitude of these changes, specific values or ranges are provided in the subsequent results section. Conclusion: In conclusion, the combined use of surgical anesthesia with DEX demonstrates a modulatory effect on the PI3K/Akt signaling pathway. This intervention proves effective in reducing hippocampal cell apoptosis post-surgery, thereby alleviating neurological impairments and ameliorating early cognitive dysfunction in rats. Our findings underscore the potential therapeutic impact of DEX in enhancing cognitive outcomes following surgical procedures. The observed reduction in hippocampal cell apoptosis and improvement in cognitive function suggest that DEX may hold promise as a neuroprotective agent in the perioperative setting. These outcomes highlight the clinical relevance of considering DEX as an adjunctive therapy to mitigate cognitive dysfunction associated with surgery. Further investigations and clinical trials are warranted to validate and extend these findings, potentially offering a novel avenue for improving patient outcomes and advancing perioperative care strategies.

TGF-ß1/SH2B3 axis regulates anoikis resistance and EMT of lung cancer cells by modulating JAK2/STAT3 and SHP2/Grb2 signaling pathways.

The pathogenesis of lung cancer, the most common cancer, is complex and unclear, leading to limited treatment options and poor prognosis. To provide molecular insights into lung cancer development, we investigated the function and underlying mechanism of SH2B3 in the regulation of lung cancer. We indicated SH2B3 was diminished while TGF-ß1 was elevated in lung cancer tissues and cells. Low SH2B3 level was correlated with poor prognosis of lung cancer patients. SH2B3 overexpression suppressed cancer cell anoikis resistance, proliferation, migration, invasion, and EMT, while TGF-ß1 promoted those processes via reducing SH2B3. SH2B3 bound to JAK2 and SHP2 to repress JAK2/STAT3 and SHP2/Grb2/PI3K/AKT signaling pathways, respectively, resulting in reduced cancer cell anoikis resistance, proliferation, migration, invasion, and EMT. Overexpression of SH2B3 suppressed lung cancer growth and metastasis in vivo. In conclusion, SH2B3 restrained the development of anoikis resistance and EMT of lung cancer cells via suppressing JAK2/STAT3 and SHP2/Grb2/PI3K/AKT signaling cascades, leading to decreased cancer cell proliferation, migration, and invasion.

miR-378d suppresses malignant phenotype of ESCC cells through AKT signaling.

BACKGROUND: Post-resistance progress in paclitaxel (PTX) treatment remains a major challenge in tumor treatment. A high dose of PTX was used for cell lines to analyze the changes in molecular expression. The miR-378d was sharply reduced in surviving cells, but the role of miR-378d in Esophageal squamous cell carcinoma (ESCC) remained unclear. METHODS: We analyzed the relationship between miR-378d expression and the clinicopathological features of ESCC. We constructed miR-378d silent expression cell lines to study phenotypes and molecular mechanisms. RESULTS: The miR-378d expression was associated with good prognosis in patients with ESCC. miR-378d inhibition promoted chemo-resistance, monoclonal formation, EMT, migration, invasion, stemness, and metastasis of ESCC cells. miR-378d can target downregulated AKT1. CONCLUSIONS: Therefore, miR-378d expression is a good prognostic factor of patients with ESCC and regulates the malignant phenotype of tumor cells through AKT.

Optimization of combined microwave and hot air drying technology for purple cabbage by Response Surface Methodology (RSM).

Purple cabbage is one of the world's most widely consumed vegetables with high nutritional values containing the antioxidants and anti-inflammatory activity of anthocyanins, vitamins, and minerals. But due to rapid postharvest quality decay, novel processing techniques including drying are required. In order to determine the conditions of combined microwave and hot air drying for purple cabbage, factors affecting the drying process including microwave density, hot air temperature, and the dry base water content at conversion point were investigated using the anthocyanin content, DPPH antioxidant capacity, chewiness, △E, rehydration ratio, and average drying rate as responses. The combined drying conditions were optimized considering three independent variables at three different levels by response surface methodology. The results showed that the processing parameters of purple cabbage with combined microwave and hot air drying technology were microwave density at 2.5 W/g, moisture content of conversion point at 4.0 g/g, and hot air temperature at 55°C. Under these conditions, the anthocyanin content, DPPH antioxidant capacity, chewiness, △E, rehydration ratio, average drying rate, and overall score of the dried purple cabbage were 175.87 mg/100 g, 87.59%, 4,521.468 g, 26.5, 4.3, 0.76 g/min, and 0.785, respectively. Therefore, combined microwave and hot air drying technology is an effective, suitable method for drying purple cabbage.

Modification of corn starch via innovative contactless thermal effect from induced electric field.

Growing attention has been focused on modifications of starch using electric field, but electrode corrosion and metal contamination remain unavoidable during the process. To solve these problems, the magneto-induced electric field was used to assist corn starch hydrolysis due to its thermal effect. Results indicated that the method accelerated corn starch acid hydrolysis and decreased the treatment time. The reducing sugar content increased to 0.59 g/L after a 60 s treatment, which was 353.44 % higher than the 20 s treatment, while the average degree of polymerization reached a minimum. The treated starch showed increased solubility and swelling power, as well as decreased freeze-thaw stability. X-ray diffraction, fourier transform infrared spectroscopy, and scanning electron microscopy results suggested that the physicochemical changes of corn starch were due to the thermal effect of the induced electric field. This study is expected to provide an important basis for applying new electric field hydrolysis technology to starch modification.

LncRNA ST7-AS1, by regulating miR-181b-5p/KPNA4 axis, promotes the malignancy of lung adenocarcinoma.

BACKGROUND: Growing evidence suggests that suppressor of tumorigenicity 7 antisense RNA 1 (ST7-AS1) is an oncogenic long noncoding RNA (lncRNA). However, little is known on its clinical significance, biological functions, or molecular mechanisms in lung adenocarcinoma (LUAD). METHODS: The expression of ST7-AS1 and miR-181b-5p were examined by qRT-PCR. The correlations between ST7-AS1 level and different clinicopathological features were analysed. In vitro, LUAD cells were examined for cell viability, migration and invasion by MTT, wound healing and Transwell assay, respectively. Epithelial-mesenchymal transition (EMT) biomarkers were detected by Western blot. The regulations between ST7-AS1, miR-181b-5p, and KPNA4 were examined by luciferase assay, RNA immunoprecipitation, RNA pulldown. Both gain- and loss-of-function strategies were used to assess the importance of different signalling molecules in malignant phenotypes of LUAD cells. The in vivo effect was analysed using the xenograft and the experimental metastasis mouse models. RESULTS: ST7-AS1 was upregulated in LUAD tissues or cell lines, correlated with tumours of positive lymph node metastasis or higher TNM stages, and associated with shorter overall survival of LUAD patients. ST7-AS1 essentially maintained the viability, migration, invasion, and EMT of LUAD cells. The oncogenic activities of ST7-AS1 were accomplished by sponging miR-181b-5p and releasing the suppression of the latter on KPNA4. In LUAD tissues, ST7-AS1 level positively correlated with that of KPNA4 and negatively with miR-181b-5p level. In vivo, targeting ST7-AS1 significantly inhibited xenograft growth and metastasis. CONCLUSIONS: ST7-AS1, by regulating miR-181b-5p/KPNA4 axis, promotes the malignancy of LUAD cells. Targeting ST7-AS1 and KPNA4 or up-regulating miR-181b-5p, therefore, may benefit the treatment of LUAD.

Effect of ropivacaine on peripheral neuropathy in streptozocin diabetes-induced rats through TRPV1-CGRP pathway.

Objective To determine the effect of ropivacaine on peripheral neuropathy in diabetic rats and its possible mechanism. Methods Forty-eight Sprague-Dawley rats were randomly divided into six groups: nondiabetic control group, nondiabetic group A (0.25% ropivacaine), nondiabetic group B (0.75% ropivacaine), diabetic control group (diabetic peripheral neuropathy (DPN) +artificial cerebrospinal fluid), diabetic group A (DPN+0.25% ropivacaine), and diabetic group B (DPN + 0.75% ropivacaine), with eight rats in each group. Within an hour of the last administration, the sciatic motor nerve conduction velocity (MNCV) of each group was measured, and the morphological changes of rat sciatic nerve were observed by HE, Weil's staining and electron microscopy. The expression of transient receptor potential vanilloid (TRPV1) in the spinal cord dorsal horn of rats was analyzed by immunohistochemistry, and the expression of Calcitonin gene-related peptide (CGRP) protein in the spinal cord was analyzed by Western blot. Results Compared with the nondiabetic control group, elevated blood glucose, decreased weight and reduced average mechanical withdrawal threshold (MWT), additionally, the sciatic nerves showed significantly slowed conduction velocity (both P<0.001) and damaged pathological structure, the expression of TRPV1 and CGRP were decreased (both P<0.001) in the diabetic groups. Compared with the diabetic control group, down-regulation of TRPV1 and CGRP in spinal cord was significant for the diabetic groups A and B treated with 0.25 and 0.75% ropivacaine, the higher concentration of ropivacaine correlated with a greater change. Conclusion Ropivacaine can significantly block sciatic nerve conduction velocity in DPN rats in a concentration-dependent manner, which may be related to the expression of the TRPV1-CGRP pathway.

Observation of clinical efficacy of application of enhanced recovery after surgery in perioperative period on esophageal carcinoma patients.

PURPOSE: To observe the clinical efficacy of the application of enhanced recovery after surgery (ERAS) in the perioperative period of esophageal carcinoma patients. METHODS: A total of 114 patients who were admitted to Affiliated Hospital of Jining Medical University for surgical treatment of esophageal carcinoma between June 2012 and June 2016 were enrolled and randomly divided into the intervention group and the regular group according to the difference of management procedures during the perioperative period. ERAS was carried out in 57 patients in the intervention group, while conventional management procedures were applied in 57 patients in the regular group. Thereafter, compared were the fluctuations in nutritional indicators and immunological indicators, postoperative complications, time to recovery of gastrointestinal function, length of stay (LOS) in hospital and cost of patients between the two groups. RESULTS: Seven days post-operation in the intervention group, the evaluation indexes of nutrition status, including total protein (TP), albumin (ALB), prealbumin (PA) and transferrin (TF), and of immunological functions, including immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM) and total blood lymphocyte count (TLC), were significantly higher than those in the regular group. As for postoperative complications, the incidence rate of the intervention group was remarkably lower than that of the regular group; the recovery time of gastrointestinal function in the intervention group was shorter than that in the regular group; the LOS in the intervention group was also shorter than that in the regular group; the in-hospital cost in the intervention group was also lower than that in the regular group. All differences above were statistically significant (p<0.05). CONCLUSION: During the perioperative period of esophageal carcinoma patients, ERAS should be fully applied to sustain the good status, and promote the recovery of immunological functions and gastrointestinal functions; at the same time, ERAS also reduces the incidence rate of postoperative complications, LOS and in-hospital cost, and we maintain that ERAS should be performed in clinical practice.

RasGRP3, a Ras guanyl releasing protein 3 that contributes to malignant proliferation and aggressiveness in human esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is one of the most common cancers worldwide; however, clinical and pathological parameters have limited ability in discriminating between clinically significant and indolent ESCC. Since RasGRP3 transcript levels have prognostic value in discriminating ESCC with different clinical aggressiveness, we decided to investigate its putative oncogenic role in ESCC. We found that RasGRP3 was highly expressed in ESCC cells. Suppression of endogenous RasGRP3 expression in esophageal cell lines reduced Ras-GTP formation as well as AKT phosphorylation. RasGRP3 suppression also inhibited cell invasion and migration and reduced proliferation, demonstrating the importance of RasGRP3 for the transformed phenotype of melanoma cells. Suppression of RasGRP3 expression in these cells inhibited downstream RasGRP3 responses and suppressed cell growth and migration, confirming the functional role of RasGRP3 in the altered behaviour of these cells. This suggests that RasGRP3 may function as a Ras activator in the phosphoinositide signalling pathway and may potentially serve as a new therapeutic target.

A comparative study on the efficacy of fast-track surgery in the treatment of esophageal cancer patients combined with metabolic syndrome.

The purpose of our study was to evaluate the clinical efficacy of fast-track surgery (FTS) in the treatment of esophageal cancer patients combined with metabolic syndrome. Ninety-four esophageal cancer patients with metabolic syndrome were selected in Affiliated Hospital of Jining Medical University from March, 2016 to February, 2017. Patients were randomly divided into control group and observation group with 47 cases in each group. Patients in observation group were treated with FTS, while patients in control group were treated with traditional method. Intraoperative blood loss, the number of dissected lymph nodes, operation time, postoperative hospital stay, the cost of hospitalization, postoperative readmission rate, and incidence of postoperative complications were compared between the groups. Levels of serum inflammatory cytokines (TNF-α and hs-CRP), fat cell factor chemerin and leptin (LP) were detected by enzyme-linked immunosorbent assay (ELISA) at 1 month after surgery. Levels of serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) at 1 month after surgery were compared between groups. Levels of hemoglobin (Hb), albumin (Alb), prealbumin (PAB) and transferrin (TRF) at 1 month after surgery were also compared between the two groups. Treatment of cancer quality-of-life questionnaire-esophageal cancer (OES-18) module was used to evaluate the symptoms of patients at one month after surgery. It turned out that no significant differences in intraoperative blood loss, operation time and the number of dissected lymph nodes were found between groups (p>0.05). Postoperative hospital stay, the cost of hospitalization, postoperative readmission rate and the incidence of postoperative complications were significantly lower in observation group than in control group (p<0.05). Levels of TNF-α, hs-CRP, chemerin and LP in observation group were significantly lower than those in control group at one month after surgery (p<0.05). Levels of TC, TG and LDL-C were significantly lower and HDL-C level was significantly higher in observation group than in control group at one month after surgery (p<0.05). Levels of Hb and Alb were significantly lower and levels of PAB and TRF were significantly higher in observation group than in control group at one month after surgery (p<0.05). OES-18 score of observation group was significantly better than that of control group at one month after surgery (p<0.05). As a conclusion, FTS can promote postoperative rehabilitation, shorten hospital stay, reduce economic burden and reduce the rehospitalization rate of esophageal cancer patients. At the same time, FTS can also improve the lipid metabolism, nutritional status and regulate the differentiation of adipocytes, alleviate the low inflammatory response state, which in turn promotes metabolic syndrome.

Increased Expression of TBP/TFIID after spinal cord injury in adult rats.

Transcription factor IID (TFIID), as a general transcription factor, plays a pivotal role in the preinitiation complex (PIC) assembly and transcription initiation by recruiting RNA polymerase II to the promoter. The TFIID complex contains the TATA-box binding protein (TBP) and a group of conserved TAF proteins. However, its distribution and function in the central nervous system (CNS) are more diverse than previously understood. Here, we mainly investigated the spatiotemporal expression and cellular localization of TBP/TFIID during spinal cord injury (SCI) in adult rats. Western blot analysis revealed that TBP/TFIID was present in normal rat's spinal cord. It gradually increased, reached a peak at the third day after SCI, and then decreased. We observed that TBP/TFIID was widely distributed in spinal cord, mainly in neurons and glial cells. In addition, Western blot detection also showed that the third day post-injury was the proliferation peak indicated by the elevated expression of proliferating cell nuclear antigen (PCNA), a marker of proliferating cells. Importantly, injury-induced expression of TBP/TFIID was colabelled by PCNA showed the increase of TBP/TFIID expression in proliferating astrocytes and microglia. Collectively, we hypothesize that TBP/TFIID may be implicated in the proliferation of astrocytes and microglia and the recovery of neurological outcomes.

Spatiotemporal expression of SKIP after rat sciatic nerve crush.

Ski-interacting protein (SKIP) is a highly conserved protein from yeast to Human. As an essential spliceosomal component and transcriptional co-regulator it plays an important role in preinitiation, splicing and polyadenylation. SKIP can also combine with Ski to overcome the G1 arrest and the growth-suppressive activities of pRb. Furthermore SKIP has the capacity to augment TGF-ß dependent transcription. While the distribution and function of SKIP in peripheral nervous system lesion and regeneration remain unclear. Here, we investigated the spatiotemporal expression of SKIP in an acute sciatic nerve crush model in adult rats. Western Blot analysis revealed that SKIP was expressed in normal sciatic nerves. It gradually increased, reached a peak at 1 week after crush, and then returned to the normal level at 4 weeks. Besides, we observed that up-regulation of SKIP was approximately in parallel with Proliferating cell nuclear antigen (PCNA), and numerous Schwann cells (SCs) expressing SKIP were PCNA and Ki-67 positive. Collectively, we hypothesized peripheral nerve crush induced up-regulation of SKIP in the sciatic nerve, which was associated with SCs proliferation.

The role of IL-6 in bone marrow (BM)-derived mesenchymal stem cells (MSCs) proliferation and chondrogenesis.

Rheumatoid arthritis (RA) is the most common degenerative arthritic cartilage and represents a disease where the prospect of stem cell therapy offers considerable hope. Currently, bone marrow (BM) represents the major source of mesenchymal stem cells (MSCs) for cell therapy. In the pathology of RA, the pro-inflammatory cytokines, such as interleukin 6 (IL-6) play a pivotal role. To investigate the direct role of IL-6 in the chondrogenic differentiation of murine MSCs (mMSCs), we isolate MSCs from the murine bone marrow, and induce MSCs chondrogenesis with different concentrations of IL-6 in vitro. Through detecting the histological and histochemical qualities of the aggregates, we demonstrate that IL-6 inhibited the differentiation of MSCs into chondrocytes in the dose-dependence manner. These findings suggest that possible strategies for improving the clinical outcome of cartilage repair procedures.

Expression of G-protein-coupled receptor kinase 6 (GRK6) after acute spinal cord injury in adult rat.

Spinal cord injury frequently results in permanent loss of neurological function. It includes many complex molecular and biochemical mechanisms. G-protein-coupled receptor kinase 6 (GRK6) is an intracellular kinase that regulates the sensitivity of certain G-protein-coupled receptors. Some studies reported GRK2 and GRK5 modulate the NFκB pathway in macrophages. Additionally, GRK2 is referred to as regulating activation of spinal cord microglia and GRK6 expression is significantly elevated in most brain regions in the MPTP-lesioned parkinsonian monkeys. However, the expression and function of GRK6 in nervous system lesion and repair are not well understood. In this study, we performed an acute spinal cord injury (SCI) model in adult rats. Western blot analysis showed the expression of GRK6 was upregulated significantly at protein level in spinal cord after SCI. Immunohistochemistry and immunofluorescence revealed wide expression of GRK6 in the normal spinal cord. After injury, GRK6 expression was increased predominantly in microglia, which expressed F4/80 (marker of macrophages and activated microglia) strongly. To understand whether GRK6 played a role in microglia activation, we applied lipopolysaccharide (LPS) to induce microglia activation in vitro. Western blot analysis demonstrated up-regulation in GRK6 protein expression after LPS stimulation was time- and dose-dependent and that up-regulation in F4/80 expression was concomitant with GRK6. These data suggested that GRK6 might be involved in the pathophysiology of SCI.

Changes in the BAG1 expression of Schwann cells after sciatic nerve crush.

Bcl-2-associated athanogene-1 (BAG1), a co-chaperone for Hsp70/Hsc70, is a multifunctional protein, which has been shown to suppress apoptosis and enhance neuronal differentiation. However, the expression and roles of BAG1 in peripheral system lesions and repair are still unknown. In this study, we investigated the dynamic changes in BAG1 expression in an acute sciatic nerve crush model in adult rats. Western blot analysis revealed that BAG1 was expressed in normal sciatic nerves. BAG1 expression increased progressively after sciatic nerve crush, reached a peak 2 weeks post-injury, and then returned to the normal level 4 weeks post-injury. Spatially, we observed that BAG1 was mainly expressed in Schwann cells and that BAG1 expression increased in Schwann cells after injury. In vitro, we found that BAG1 expression increased during the cyclic adenosine monophosphate (cAMP)-induced Schwann cell differentiation process. BAG1-specific siRNA inhibited cAMP-induced Schwann cell differentiation. In conclusion, we speculated that BAG1 was upregulated in the sciatic nerve after crush, which was associated with Schwann cell differentiation.

FOXO3a/p27kip1 expression and essential role after acute spinal cord injury in adult rat.

FOXO3a (Forkhead Class box O3a), as an important direct target of the phosphatidylinositol 3-kinase (PI3K)/protein B (Akt) pathway, which regulates the cell survival and the cell-cycle progression. Recent reports showed that FOXO3a could inhibit cell-cycle progression at the G1/S transition by controlling transcription of the cyclin-dependent kinase inhibitor p27(kip1) , which is also a key regulator of the mammalian neurogenesis. To elucidate the expression and role of FOXO3a in nervous system lesion and repair, we performed an acute spinal cord contusion injury (SCI) model in adult rats, which showed a temporal-spatial expression pattern of FOXO3a. Temporally, FOXO3a protein level significantly reduced day 3 after injury, and following FOXO3a down-regulation, p27(kip1) protein and mRNA levels were also decreased after injury. Spatially, decreased levels of FOXO3a and p27(kip1) were predominant in astrocytes, which were regenerating axons and largely proliferated after injury. Furthermore in vitro, Western blot analysis, RT-PCR, and immunofluorescence staining analysis demonstrated the relationship between FOXO3a and p27(kip1) in primary astrocytes. FOXO3a modulated the cell cycle by transcriptional regulation of p27(kip1) in astrocytes. Administration of the PI3K pharmacological inhibitor LY294002 abrogated this effect by regulating FOXO3a and p27(kip1) expression and subcellular localization. These results suggest that decreased levels of FOXO3a and p27(kip1) in spinal cord are involved in axonal regeneration and the proliferation of glial cells after SCI.

An upregulation of SENP3 after spinal cord injury: implications for neuronal apoptosis.

SENP3 (SUMO-specific proteases 3), a member of the small ubiquitin-like modifier specific protease family, was identified as a molecule that deconjugates SUMOylation of modified protein substrates and functions as an isopeptidase by disrupting SUMO homeostasis to facilitate cancer development and progression. However, its expression and function in nervous system injury and repair are still unclear. In this study, we employed an acute spinal cord injury (SCI) model in adult rats and investigated the dynamic changes of SENP3 expression in the spinal cord. Western blot analysis indicated a gradual increase in SENP3 expression, which peaked 3 days after SCI, and then declined over the following days. Immunohistochemistry results further confirmed that SENP3 was expressed at low levels in the gray and white matter in the non-injured condition and increased after SCI. Moreover, immunofluorescence double-labeling showed that SENP3 was co-expressed with the neuronal marker, NeuN. Furthermore, the SENP3-positive cells that were co-expressed with NeuN had also expressed active caspase-3 after injury. To investigate whether SENP3 plays a role in neuronal apoptosis, we applied H(2)O(2) to induce neuronal apoptosis in vitro. Western blot analysis showed a significant upregulation of SENP3 and active caspase-3 following H(2)O(2) stimulation. Taken together, these results suggest that SENP3 may play important roles in the pathophysiology of SCI.

Changes in CLIP3 expression after sciatic nerve injury in adult rats.

CLIP3 (cytoplasmic linker protein 3) is a 547 amino acid residue cytoplasmic protein that localises to Golgi stacks and tubulovesicular elements juxtaposed to Golgi cisternae. Composed of three Ank (ankyrin) repeats and two CAP-Gly (cytoskeleton-associated protein-glycine) domains, CLIP3 may function as a cytoplasmic linker protein that is involved in TGN-endosome dynamics. To define the expression and role of CLIP3 during peripheral nervous system degeneration and regeneration, we created an acute sciatic nerve injury (SNI) model in adult rats. Western blot analyses revealed prominent up-regulation of CLIP3 and PCNA (proliferating cell nuclear antigen) protein levels at 3 days after SNI. Immunohistochemistry displayed that the expression of CLIP3 was noticeably increased in the injured nerve. Immunofluorescence further revealed that the CLIP3 and PCNA proteins colocalised respectively with S100 in the cytoplasm of Schwann cells. The expression profile of the SC/neuron co-cultures demonstrated that CLIP3 and PCNA protein levels were markedly expressed during the early stage of myelination. These results suggest that CLIP3 is likely associated with the myelination of proliferating Schwann cells, and nerve tissue regeneration after peripheral nerve injury. CLIP3 and PCNA expression during early myelination may be related to the direct uptake and transport of lipids and cholesterol, which were derived from the degenerating myelin, by Schwann cells to prepare for the formation of myelin sheath-like structures around regenerated axons after SNI.

Spatiotemporal patterns and essential role of TNF receptor-associated factor 5 expression after rat spinal cord Injury.

TRAF5 (TNF receptor-associated factor 5), which has a tumor necrosis factor receptor-associated factor (TRAF) domain in its carboxyl terminus, was identified CD40-associated factor. It was a signal transducer for NF-κB signal pathway and other pathway. To elucidate the expression and roles of TRAF5 in nervous system lesion and repair, we performed an acute spinal cord injury (SCI) model in adult rats and studied the dynamic changes of TRAF5 expression in spinal cord. Western blot and immunohistochemistry analysis revealed that TRAF5 was present in normal spinal cord. It gradually increased, reached a peak at 5 days after SCI, and then declined during the following days. Immunofluorescence double-labeling revealed that TRAF5 was co-expressed with NeuN and GFAP, respectively. Interesting, after injury, TRAF5 expression was increased predominantly in astrocytes, which highly expressed PCNA, a marker for proliferating cells. In conclusion, this is the first description of TRAF5 expression in spinal cord. Our data suggested that TRAF5 might play important roles in CNS pathophysiology after SCI.