Staining For Neuron-specific Enolase Research Articles

Pharmacodynamics and mechanism of Erigeron breviscapus granules in the treatment of ischemic stroke in mice by regulating sphingolipid metabolism based on metabolomics

AimErigeron breviscapus (Vant.) Hand.-Mazz. (EB) granules is the extract preparation of EB, with clear curative effect and unclear mechanism. This study intends to systematically explore the specific mechanism of EB granules in the treatment of IS from the metabolic perspective. MethodsThe model of transient middle cerebral artery occlusion (tMCAO) in mice was established by the suture-occluded method. The therapeutic effect of EB granules on tMCAO mice was evaluated by behavioral evaluation, brain water content determination, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, and levels of lactate dehydrogenase (LDH) and neuron specific enolase (NSE) in serum. In order to screen differential metabolites, non-targeted metabolomics technology was used to detect the metabolites in serum before and after administration. Univariate statistics, multivariate statistics and bioinformatics were used to analyze the changes of metabolites in serum of tMCAO mice. The possible related mechanism of EB granules in treating IS was screened by pathway enrichment analysis, and the preliminary verification was carried out at animal level by enzyme linked immunosorbent assay (ELISA) and western blot (WB). ResultsEB granules could significantly improve behavior of tMCAO mice, reduce brain water content and cerebral infarction volume, improve morphology of brain tissue, reduce the levels of LDH and NSE in serum. A total of 232 differential metabolites were screened, which were mainly enriched in many biological processes such as sphingolipid metabolism. The differential metabolite S1P and its receptors S1PR1 and S1PR2 in sphingolipid metabolism were verified. The results showed that the level of S1P in brain tissue increased and the protein expression of S1PR1 decreased significantly after modeling, and reversed after administration, but there was no significant difference in the protein expression of S1PR2. ConclusionThe therapeutic effects of EB granules may be related to affecting sphingolipid metabolism through regulating S1P/S1PR1.

Investigating the role of Purkinje fibers and synaptic connectivity in balance regulation through comprehensive ultrastructural and immunohistochemical analysis of the donkey's (Equus asinus) cerebellum.

The donkey's extraordinary capacity to endure substantial loads over long distances while maintaining equilibrium suggests a distinctive cerebellar architecture specialized in balance regulation. Consequently, our study aims to investigate the intricate histophysiology of the donkey's cerebellum using advanced ultrastructural and immunohistochemical methodologies to comprehend the mechanisms that govern this exceptional ability. This study represents the pioneering investigation to comprehensively describe the ultrastructure and immunohistochemistry within the donkey cerebellum. Five adult donkeys'cerebella were utilized for the study, employing stains such as hematoxylin, eosin, and toluidine blue to facilitate a comprehensive histological examination. For immunohistochemical investigation, synaptophysin (SP), calretinin, and glial fibrillary acidic proteinwere used and evaluated by the Image J software. Furthermore, a double immunofluorescence staining of SP and neuron-specific enolase (NSE) was performed to highlight the co-localization of these markers and explore their potential contribution to synaptic function within the donkey cerebellum. This investigation aims to understand their possible roles in regulating neuronal activity and synaptic connectivity. We observed co-expression of SP and NSE in the donkey cerebellum, which emphasizes the crucial role of efficient energy utilization for motor coordination and balance, highlighting the interdependence of synaptic function and energy metabolism. The Purkinje cells were situated in the intermediate zone of the cerebellum cortex, known as the Purkinje cell layer. Characteristically, the Purkinje cell's bodies exhibited a distinct pear-like shape. The cross-section area of the Purkinje cells was 107.7 ± 0.2 µm2 , and the Purkinje cell nucleus was 95.7 ± 0.1 µm2 . The length and diameter of the Purkinje cells were 36.4 × 23.4 µm. By scanning electron microscopy, the body of the Purkinje cell looked like a triangular or oval with a meandrous outer surface. The dendrites appeared to have small spines. The Purkinje cells'cytoplasm was rich with mitochondria, rough endoplasmic reticulum, ribosomes, Golgi apparatus, multivesicular bodies, and lysosomes. Purkinje cell dendrites were discovered in the molecular layer, resembling trees. This study sheds light on the anatomical and cellular characteristics underlying the donkey's exceptional balance-maintaining abilities.

Diclofenac sodium and dexamethasone co-therapy restores brain neuron-specific enolase (NSE), S-100 Beta and glial fibrillary acid protein (GFAP) proteins in experimental rat's model: A possible inhibition of P-glycoprotein.

Non-steroidal anti-inflammatory drugs decrease pain and fever while corticosteroids regulate inflammation and immune response, both are prescribed to reduce inflammation and control pain. The present study aimed to study the effects of their monotherapy and co-administration on the brain tissue structure of experimental rats. P-glycoprotein (PGP), a transporter membrane protein, plays an important role in various physiological and physio-pathological conditions, drug-drug and drug-food interactions, and multi-drug resistance. Male rats were divided into four groups and received normal saline, dexamethasone, diclofenac sodium and their dual therapy respectively, then after one-month rats were sacrificed and brain tissues proceeded for hematoxylin and eosin staining to study their histopathology and immunohistochemically staining of NSE, S100-B and GFAP biomarkers were performed. Additionally, in silico molecular docking studies were conducted to elucidate interactions between PGP and used compounds. Resultsshowed that dexamethasone or diclofenac sodium treatments showed abnormalities like edema, neuronal vacuoles, astrocytes hyperplasia and microglial cells with positive reaction to NSE, S100 and GFAP antibodies while the dual therapy displayed less edema and other signs of damage with negative and weak positive staining of NSE, S100 and GFAP antibodies respectively. The molecular docking showed that there were different affinities toward the involved PGP active site. These interaction results were great with Dexamethasone -9.6 kcal/mol forming hydrophobic interactions with the highest affinity when compared with Diclofenac sodium which gave -8.4 kcal/mol. In conclusion, the side effects of the two types of anti-inflammatory drugs may be minimized through their interactions. However, Molecular Dynamic Simulations studies are required to explain the exact dynamic behaviors and protein-ligand stability.

Transcranial photobiomodulation (808nm) attenuates pentylenetetrazole-induced seizures by suppressing hippocampal neuroinflammation, astrogliosis, and microgliosis in peripubertal rats.

.Significance: Transcranial photobiomodulation (tPBM) at 808 nm attenuates pentylenetetrazole (PTZ)-induced seizures and convulsive status epilepticus (CSE) in peripubertal rats by protecting neurons from injury and parvalbumin-positive interneurons from apoptosis, and preserving the integrity of perisomatic inhibitory networks. However, the effects of tPBM on neuroinflammation, astrogliosis, and microgliosis in epileptic rat brains are unknown. Thus, further study to unveil these aspects is needed for understanding the phenomena of tPBM on pediatric CSE prevention.Aim: To evaluate the effects of tPBM on neuroinflammation, astrogliosis, and microgliosis in peripubertal rat hippocampus with PTZ-induced seizures and SE.Approach: An 808-nm diode laser was applied transcranially to peripubertal rats prior to PTZ injection. Immunofluorescence staining of neuron-specific enolase (NSE) was used as a marker of neuroinflammation, glial fibrillary acid protein (GFAP) for astrogliosis, ionized calcium-binding adapter molecule 1 (Iba-1) for microgliosis, and mitochondrial cytochrome c oxidase subunit 1 (MT-CO1) for confirming the involvement of cytochrome c oxidase (CCO).Results: tPBM significantly reduced NSE immunoreactivity in CA3 in PTZ-treated rats, GFAP immunoreactivity in CA1, and Iba-1 immunoreactivity in CA3. Enhancement of hippocampal MT-CO1 reflected that tPBM acted in CCO-dependent manner.Conclusions: tPBM (808) attenuated PTZ-induced seizures and SE by suppressing neuroinflammation, astrogliosis, and microgliosis in peripubertal rats.

Prognostic value of the immunohistochemistry markers CD56, TTF-1, synaptophysin, CEA, EMA and NSE in surgically resected lung carcinoid tumors.

Lung carcinoid tumor is a type of neuroendocrine tumor, which is subdivided into typical carcinoid (TC) and atypical carcinoid (AT), based on the rate of mitosis and the presence of necrosis. Several prognostic factors for lung carcinoids have been reported in the literature, including the type, Ki67 index, stage, chemotherapy and radiation therapy. In the present study, 108 cases with resected carcinoid lung tumors were enrolled and the expression of CD56, thyroid transcription factor 1, synaptophysin, carcinoembryonic antigen, epithelial membrane antigen and neuron-specific enolase (NSE) in the resected tissue specimens was immunohistochemically analyzed. Patients with positive staining for NSE had an unfavorable survival prognosis compared with patients with negative staining for NSE (137.2 vs. 150.0 months, P=0.044). According to univariate analysis, none of the above immunohistochemistry markers was associated with survival, and according to multivariate analysis, NSE was an independent influencing factor for survival inpatients with AT (P=0.046) and furthermore, the stage was an independent factor of survival in patients with TC (P=0.005).

Surgical Treatment of Primary Cardiac Paragangliomas: A Single-Center Experience.

Cardiac paragangliomas are rare neuroendocrine tumors that will cause significant morbidity if left undiagnosed. Because of the paucity of cohort data, their rapid diagnosis and appropriate management still pose unique challenges to cardiac surgeons. We aimed to investigate the clinical features and surgical management of primary cardiac paragangliomas in our single center. From May 2014 to October 2020, patients diagnosed with primary cardiac paragangliomas retrospectively were reviewed. Demographic data, clinical presentation, preoperative imaging methods, surgical resection, perioperative management, histological analysis, and outcomes were recorded. Postoperative follow up also was reviewed. With multiple imaging methods, including echocardiography, computed tomography, positron-emission tomographic-computed tomography, and biochemical tests, there were five cases of primary cardiac paraganglioma verified by postoperative immunohistochemical staining, two of which were hormonally active. There were no metastatic cardiac paragangliomas, according to positron-emission tomographic-computed tomography, and all patients accepted surgical treatment. Preoperative adrenoceptor blockade was given to hormonally active patients, accordingly. Complete resection of the tumor was accomplished under cardiopulmonary bypass in each case. Tumor distribution included two masses on the roof of the left atrium, two masses in the right atrioventricular groove, and one between the ascending aorta and main pulmonary artery. Immunohistochemical staining for chromogranin, neuron-specific enolase, synaptophysin, and S-100 were positive, which were typical of cardiac paraganglioma. There were no operative deaths. All the patients had an uneventful recovery except one patient who underwent low cardiac output syndrome. During follow up (mean 4.2 years, range 0.6-7.0 years), all patients were well with New York Heart Association class I or II. Only one patient developed thyroid carcinoma three years after surgery but with no paraganglioma recurrence during periodic computed tomography, and this patient recovered well after thyroidectomy. Although cardiac paragangliomas are rare and may present surgical challenges for clinicians, surgical resection remains the choice of treatment with favorable outcomes through a multidisciplinary heart team. Moreover, lifelong surveillance still is recommended to detect possible recurrence or associated nonchromaffin tumors in time.

Sevoflurane Inhibits Traumatic Brain Injury-Induced Neuron Apoptosis via EZH2-Downregulated KLF4/p38 Axis

Traumatic brain injury (TBI) is characterized by physical damage to the brain tissues, ensuing transitory or permanent neurological dysfunction featured with neuronal loss and subsequent brain damage. Sevoflurane, a widely used halogenated anesthetic in clinical settings, has been reported to alleviate neuron apoptosis in TBI. Nevertheless, the underlying mechanism behind this alleviation remains unknown, and thus was the focus of the current study. First, Feeney models were established to induce TBI in rats. Subsequently, evaluation of the modified neurological severity scores, measurement of brain water content, Nissl staining, and TUNEL assay were employed to investigate the neuroprotective effects of sevoflurane. Immunofluorescence and Western blot analysis were further applied to detect the expression patterns of apoptosis-related proteins as well as the activation of the p38-mitogen-activated protein kinase (MAPK) signaling pathway within the lesioned cortex. Additionally, a stretch injury model comprising cultured neurons was established, followed by neuron-specific enolase staining and Sholl analysis. Mechanistic analyses were performed using dual-luciferase reporter gene and chromatin immunoprecipitation assays. The results demonstrated sevoflurane treatment brought about a decrease blood-brain barrier (BBB) permeability, brain water content, brain injury and neuron apoptosis, to improve neurological function. The neuroprotective action of sevoflurane could be attenuated by inactivation of the p38-MAPK signaling pathway. Mechanistically, sevoflurane exerted an inhibitory effect on neuron apoptosis by up-regulating enhancer of zeste homolog 2 (EZH2), which targeted Krüppel-like factor 4 (KLF4) and inhibited KLF4 transcription. Collectively, our findings indicate that sevoflurane suppresses neuron apoptosis induced by TBI through activation of the p38-MAPK signaling pathway via the EZH2/KLF4 axis, providing a novel mechanistic explanation for neuroprotection of sevoflurane in TBI.

Methane Saline Ameliorates Traumatic Brain Injury through Anti-Inflammatory, Antiapoptotic, and Antioxidative Effects by Activating the Wnt Signalling Pathway.

Objective Methane saline (MS) can be used to treat many diseases via its anti-inflammatory, antiapoptotic, and antioxidative activities. However, to date, there is no published evidence as to whether MS has any effect on traumatic brain injury (TBI). The Wnt signalling pathway regulates cell proliferation, differentiation, migration, and apoptosis; however, whether the Wnt signalling pathway regulates any effect of MS on TBI is unknown. This study was designed to explore the role of MS in the treatment of TBI and whether the Wnt pathway is involved. Methods Sprague-Dawley rats were randomly divided into five groups: sham, TBI, TBI+10 ml/kg MS, TBI+20 ml/kg MS, and TBI+30 ml/kg MS. After induction of TBI, MS was injected intraperitoneally once daily for seven consecutive days. Neurological function was evaluated by the Neurological Severity Score (NSS) at 1, 7, and 14 days after TBI. Haematoxylin-eosin (HE) staining, inflammatory factors, neuron-specific enolase (NSE) staining, oxidative stress, and cell apoptosis were measured and compared 14 d after TBI to identify the optimal dose of MS and to investigate the effect of MS on TBI. In the second experiment, Sprague-Dawley rats were randomly divided into four groups: sham, TBI, TBI+20 ml/kg MS, and TBI+20 ml/kg MS+Dickkopf-1 (DKK-1, a specific inhibitor of the Wnt pathway). NSE, caspase-3, superoxide dismutase (SOD), Wnt3a, and β-catenin were detected by real-time PCR and Western blotting. The results from each group were compared 14 d after TBI to determine the regulatory role of the Wnt pathway. Results Methane saline significantly inhibited inflammation, oxidative stress, and cell apoptosis, thus protecting neurons within 14 days of TBI. The best treatment effect against TBI was obtained with 20 ml/kg MS. When the Wnt pathway was inhibited, the treatment effect of MS was impaired. Conclusion Methane saline ameliorates TBI through its anti-inflammatory, antiapoptotic, and antioxidative effects via activation of the Wnt signalling pathway, which plays a part but is not the only mechanism underlying the effects of MS. Thus, MS may be a novel strategy for treating TBI.

Effects of Different Target Temperatures on Angiogenesis and Neurogenesis Following Resuscitation in a Porcine Model After Cardiac Arrest.

The optimal effective temperature of targeted temperature management (TTM) used to prevent cerebral injury following cardiopulmonary resuscitation (CPR) is undetermined. In this study, we compared the mortality, neurologic deficits, and cerebral protein levels of two target temperatures. Fifty 4-month-old female domestic pigs were randomized to sham, TTM at 33°C ± 0.5°C (T33), TTM at 35°C ± 0.5°C (T35), and normothermic (NT) groups. In the NT and TTM groups, untreated ventricular fibrillation was induced electrically in animals for 10 min, followed by 6 min of CPR. Target core temperatures (Tc) of TTM groups were induced and maintained (6 h) using an endovascular hypothermia device, and rewarmed to 37.5 ± 0.5°C in the next 6 h. Tc of the NT group was maintained at 37.5 ± 0.5°C. The survival outcomes and neurological function were evaluated every 24 h for 72 h. All animals were successfully resuscitated with no significant differences in baseline characteristics or hemodynamic indexes. Survival rates and neurological outcomes were significantly improved in the TTM groups, with T33 showing the most significant effect. Compared with NT-treated animals, TTM-treated animals had higher expressions of angiopoietin-1, transforming growth factor-alpha , vascular endothelial growth factor, metallopeptidase inhibitor (TIMP)-1, TIMP-2, and platelet-derived growth factor-BB. Macrophage migration inhibitory factor and IL-17F levels were markedly upregulated after resuscitation in the NT group but inhibited in the TTM groups. Neuron-specific enolase staining data was also consistent with our conclusion that hypothermia can reduce reperfusion-induced brain injuries. Lower target temperature showed greater protective effects against cerebral injuries after CPR, and the improved neurological outcomes after TTM may be associated with decreased expression of pro-inflammatory cytokines and increased expression of blood-brain barrier and neurogenesis regulatory factors in this porcine model of CA following resuscitation.

2652 Primary Malignant Peripheral Nerve Sheath Tumor of Small Bowel and Colon Presenting With Metastases to Bone, Lung, Liver, Spleen, and Brain

INTRODUCTION: Malignant Peripheral Nerve Sheath Tumor (MPNST) is a rare diagnosis, with an incidence of 1:100,000 per year and 4–10% of soft tissue sarcoma, contains properties both neural crest cells and mesenchymal cells. Gastrointestinal MPNSTs are rare disease conditions with only 10 cases reported worldwide in the literature. CASE DESCRIPTION/METHODS: A 71-year old Portuguese male, presented with 8-week history of abdominal pain, anorexia with 20Ib weight loss. Vital signs were normal. On physical examination, a firm, tender, fixed abdominal mass in the RLQ was palpated. CT scan of the abdomen showed a 10 cm necrotic mass on the right side of the abdomen with communication to the small bowel was detected with a suggestion of a walled off perforation. Colonoscopy revealed a flattening of the terminal ileum suggesting an extrinsic compression. Biopsy of tumor consisting of mixed spindle and pleomorphic cells with focal rhabdoid features, high grade nuclear features and brisk mitotic activity. A week later, he presented with symptoms of small bowel obstruction, the CT scan at this time showed a 8 cm left sided abdominal mass with communication to the small bowel with dilation of the jejunum suggesting a new partial obstruction in addition. The patient underwent a palliative debulking of the tumor. Multiple large obstructive masses were found in the Ligament of Treitz,significant portion of the small bowel, right colon, left upper quadrant and omentum. For palliation purposes, these masses were resected and anastomosis was performed. Gross examination of the surgical specimens revealed grayish-tan cystic masses filled with brownish tan hemorrhagic material. Histology of the masses were similr to previous mass. On immunohistochemistry, showed patchy weak S100 immunoreactivity and focal BCL-2 and Neuron Specific Enolase stain immunoreactivity. Ki-67 proliferation index was 70%. The pathologic diagnosis of MPNST was established. He developed multiple bone, pulmonary, liver, splenic and hemorrhagic brain metastases. He died under hospice care within six months of his initial presentation. DISCUSSION: MPNST, occurs mostly in association with Neurofibromatosis Type 1 (8–13%). The tumor typically originates from the nerve sheaths of the major nerves in the extremities (40%), trunk/ retroperitoneal (38%) and head and neck region (21%). However, recent case reports have also documented MPNST of the liver, thyroid gland, skin, eighth cranial nerve and rarely the greater omentum without a history of radiation or NF1.

Desferrioxamine and dextromethorphan combination exhibited synergistic effect and reversed the catalepsy behaviour in 6-hydroxydopamine hydroydopamine administered rats through regulating brain glutamate levels.

To investigate the effect of desferrioxamine (DFO) and dextromethorphan (DXM) combination in animal model of Parkinson's disease (PD). The PD was induced in rats through intracerebroventricular administration of 6-hydroxydopamine (6-OHDA) using stereotaxic apparatus. The animals were subjected to behavioural assessments and neurobiochemicals estimation followed by immunohistochemistry staining of neuron specific enolase (NSE) in striatum. Desferrioxamine and DXM combination has significantly reversed the catalepsy behaviour and elevated the antioxidant enzymes (SOD, CAT, GSH) and dopamine levels. Interestingly, the level of glutamate, nitric oxide, cytokines (IL-1β, TNF-α) and NSE expressions were found to be decreased in striatum region of 6-OHDA-administered rats. The combination of DFO and DXM has shown synergism in most of the parameters studied, when compared to per se treatment. The reversal of catalepsy behaviour represents the protective effect of above combination on dopamine neurons in striatum from 6-OHDA toxicity. The mechanism of DFO and DXM combination might be attributed through attenuation of glutamate-induced excitotoxicity in neurons through ameliorating the reactive oxygen species and pro-inflammatory cytokines release. Treatment with DFO and DXM combination could control the multiple events in the pathogenesis of PD.

The relationship between serum tumor markers and the severity of silicosis

Objective To investigate the relationship between serum tumor markers and the severity of silicosis. Methods Retrospective study.Total of 160 patients with silicosis were included in the study, and 160 healthy volunteers were recruited as the control group. Tumor marker levels in both bronchoalveolar lavage fluid(BALF)and serum were detected by the immunochemiluminecence methods. The pulmonary function parameters, blood gas analysis and lactate dehydrogenase(LDH)were also analyzed. Lung tissue obtained by a patient with silicosis was stained by neuron specific enolase(NSE), carbohydrate antigen125(CA125) and carbohydrate antigen19-9(CA19-9). Results Serum NSE, CA125 and CA19-9 levels were significantly higher in cases than those in controls[(34.47±13.30)μg/L vs(10.24±7.20)μg/L, t=20.27, P<0.000 1; (33.96±17.80)kU/L vs(12.23±15.30)kU/L, t=11.71, P<0.000 1; (4.68±5.67)kU/L vs(2.78±3.45)kU/L, t=3.67, P<0.002]. Significant negative correlations were found between values of tumor markers(CA125 and CA19-9) and spirometric parameters, such as forced expiratory volume in one second %(FVE1%), forced expiratory volume in one second/forced vital capacity(FVE1/FVC), carbon monoxide diffusion capacity (Dlco) and total lung capacity(TLC) (r=-0.423, P=0.001; r=-0.323, P=0.011; r=-0.479, P=0.001; r=-0.285, P=0.043) and (r=-0.324, P=0.022; r=-0.256, P=0.023; r=-0.354, P=0.013; r=-0.356, P=0.012). Significant positive correlations were also observed between values of these tumor markers and LDH(r=0.378, P=0.001 and r=0.347, P=0.21). Significant negative correlations were found between NSE and Dlco and TLC(r=-0.374, P=0.004 and r=-0.368, P=0.002). Significant positive correlations were also observed between NSE and LDH(r=0.404, P=0.001). The NSE and CA19-9 levels in BALF were significantly higher than those in serum[(39.32±29.30)μg/L vs(25.7±12.12)μg/L, t=2.15, P=0.036; (21.36±12.11)kU/L vs(11.28±10.78)kU/L, t=2.64, P=0.012]. Patients experienced a decrease in NSE and CA19-9 concentrations following whole lung lavage[(39.20±10.24)μg/L vs(15.32±8.35)μg/L, t=8.02, P<0.05; (26.24±12.23)kU/L vs(18.84±5.64)kU/L, t=2.46, P<0.05]. Immunohistochemical studies showed positive NSE and CA19-9 staining in lung biopsy specimen. Conclusion Elevated serum tumor markers including NSE, CA125 and CA19-9 would provide valuable clinical information to assess disease severity in silicosis.(Chin J Lab Med, 2017, 40: 515-519) Key words: Silicosis; Tumor markers, biological; CA-125 antigen; CA-19-9 antigen

Gastric neuromuscular histology in patients with refractory gastroparesis: Relationships to etiology, gastric emptying, and response to gastric electric stimulation

The aims of this study were to describe the histology in gastroparesis, specifically to relate histopathology to etiology of gastroparesis (idiopathic and diabetic gastroparesis), gastric emptying, and clinical response to gastric electric stimulation. Full thickness gastric body sections obtained during insertion of gastric stimulator in gastroparetics were stained with Hematoxylin & Eosin, Masson Trichrome and immunohistochemical stains for Neuron-Specific Enolase and c-Kit. In all, 145 gastroparetics (71 diabetics, 71 idiopathic, 2 post-surgical, and 1 chronic intestinal pseudo-obstruction) had full thickness gastric body biopsies. A lymphocytic infiltrate was seen in the intermyenteric plexus in 22 diabetic and 23 idiopathic gastroparesis patients. Fibrosis was present in the inner circular layer in 13 diabetic and 15 idiopathics and in the outer longitudinal layer in 46 diabetic and 51 idiopathics. Diabetic gastroparesis had less ganglion cells (3.27±1.82 vs 4.81±2.81/hpf; P<.01) and less ganglia (0.90±0.44 vs 1.10±0.50/hpf; P=.01) than idiopathic gastroparesis. Interstitial cells of Cajal (ICC) count was slightly lower in the inner circular layer in diabetic than idiopathics (2.77±1.47 vs 3.18±1.34/hpf; P=.08). Delayed gastric emptying was associated with reduced ICCs in the myenteric plexus. Global therapeutic response to gastric electric stimulation was inversely related to ganglia/hpf (R=-.22; P=.008). In diabetics, improvements in nausea, vomiting, and abdominal pain were inversely related to fibrosis. Histologic assessment of full thickness gastric biopsy specimens allows correlation of histopathology to the gastroparesis disease process, its etiology, gastric emptying, and response to gastric electric stimulation treatment.

Effect of a new ROCK inhibitor thiazovivin on the morphology and function of human corneal endothelial cells

To evaluate the effect of thiazovivin, a novel ROCK inhibitor, on the morphology and function of human corneal endothelial cells(HCECs). The primary HCECs were identified by light microscopy and immunofluorescence staining of neuron-specific enolase. To screen the optimal concentration and action time of thiazovivin for maintaining the morphology and function of primary HCECs, Na (+)/K (+)-ATPase and N-cadherin were chosen as indicators, and the morphology and function of HCECs in various concentrations(0 μmol/L, 2 μmol/L, 4 μmol/L, and 6 μmol/L)for different durations(24 h and 48 h)were examined by immunofluorescence experiments. The effect of thiazovivin on the expression of ROCK was investigated by immunofluorescence and Western blot. The primary HCECs cultured were hexagonal, closely packed, homogeneously and obviously stained by neuron-specific enolase. The immunofluorescence staining of Na(+)/K(+)-ATPase showed that when the primary HCECs cultured with various concentrations of thiazovivin(0, 2, 4, 6 μmol/L)for 24 h, the fluorescence were obvious, and the average absorbance values(A)were 1.27±0.08, 3.72±0.17, 21.07±4.67, 3.69±0.34, respectively. And the immunofluorescence staining of N-cadherin revealed that when the primary HCECs treated with 4 μmol/L thiazovivin for 24 h, the cell boundary was clear and the structure of the cells was intact. While the treating time of thiazovivin(4 μmol/L)on HCECs extended to 48 h, the immunofluorescence staining of Na(+)/K(+)-ATPase and N-cadherin showed that compared to HCECs treated with thiazovivin(4 μmol/L)for 24 h, the fluorescence intensity did not change significantly, but the cells arranged slightly untidy. In addition, the immunofluorescence staining of ROCK was weakened and the expression of ROCK was reduced by thiazovivin. Thiazovivin was effective for protecting the morphology and function of HCECs. An optimal improvement in the morphology, connection and function of HCECs was found when the primary HCECs were cultured with 4 μmol/L thiazovivin for 24 h. Moreover, the expression of ROCK protein could be significantly inhibited by thiazovivin. (Chin J Ophthalmol, 2016, 52: 686-692).

Neuron-specific enolase as a novel biomarker reflecting tuberculosis activity and treatment response.

Background/Aims:It is not clear which tests are indicative of the activity and severity of tuberculosis (TB). This study aimed to investigate the predictive value of neuron-specific enolase (NSE) and to determine the origin of NSE in TB patients.Methods:A single-center retrospective analysis was conducted on newly diagnosed TB patients between January and December 2010. Patients were categorized into one of two disease groups (focal segmental or extensive) based on chest X-ray. Pre- and post-treatment NSE concentrations were evaluated. To determine the origin of serum NSE concentration, NSE staining was compared with macrophage-specific CD68 staining in lung tissues and with a tissue microarray using immunohistochemistry and immunofluorescence.Results:A total of 60 newly diagnosed TB patients were analyzed. In TB patients, NSE serum concentration was significantly increased and NSE level decreased after treatment (p < 0.001). In proportion to serum high-sensitivity C-reactive protein concentration, the mean serum concentration of NSE in the extensive group (25.12 ng/mL) was significantly higher than that in the focal segmental group (20.23 ng/mL, p = 0.04). Immunohistochemical staining revealed a large number of macrophages that stained positively for both NSE and CD68 in TB tissues. In addition, NSE signals mostly co-localized with CD68 signals in the tissue microarray of TB patients.Conclusions:Our results suggest that NSE may be a practical parameter that can be used to monitor TB activity and treatment response. Elevated serum NSE level originates, at least in part, from macrophages in granulomatous lesions.

Histochemical Changes in the Cerebellum of Wistar Rats Administered with Oral Doses of Zidovudine

Zidovudine is a drugs used in the management of Human Immunodeficiency Virus (HIV) and Acquired Immunodeficiency Syndrome (AIDS) infection in sub-Saharan Africa in combination with other drugs. The objective of this research was to investigate the potential harmful effects of this drug on the histology of the cerebellum of Wistar rats. Twenty male Wistar rats were used for this study. The rats were divided into 2 groups of 10 rats each. Group A served as the control and was administered with 1 ml of distill water, while group B was administered with 8.57mg/kg of zidovudine daily for 30 days, after which the rats where sacrificed and each cerebellum was harvested, processed and stained using haematoxylin and Eosin (H/E), silver impregnation method. Paraffin impregnated Glial Fibrilar Acidic Protein (GFAP), Neuron Specific Enolase (NSE) and Neurofilament (NF) immunochemistry methods. Stained slides were viewed using light microscope. Results showed that, the cerebellum of Groups B animals were affected with moderate to severe shrinking and distortion of the Purkinje cells and granular cells, when compared with the control. Group B animals, also showed more expression of GFAP, NSE and NF staining in their cerebellum than the control. This suggests that zidovudine is harmful to the cerebellum and should be taken with caution

Interval progression of idiopathic hypertrophic pachymeningitis.

A 38-year-old right-handed woman first presented at age 16 with compressive myelopathy. A C7-T2 decompressive laminectomy was performed after imaging revealed diffuse thickening of spinal and intracranial dura. Biopsy revealed lymphocytic infiltration of the dura (infiltration of the dura by a population of monomorphic small cells with scant cytoplasm and dense nuclei) with no features to suggest neoplasia. No giant cell, granuloma, or microorganisms were seen. Mitoses were not seen and negligible nuclear atypia was reported. Immunohistochemical staining for lymphocyte common antigen was positive in most of the inflammatory cells. Glial fibrillary acidic protein and neuron-specific enolase staining was not seen. Taking her history of rheumatoid arthritis, glomerulonephritis, and Hashimoto thyroiditis into consideration, after an exhaustive evaluation, a diagnosis of idiopathic hypertrophic pachymeningitis was made. She was initially maintained on prednisone and later methotrexate for about 10 years but self-discontinued it in 2005 as she had no symptoms. In February 2009, brain MRI was carried out to evaluate for occipital headaches (figure 1). Amitriptyline resulted in headache relief and she was lost to follow-up but presented again in March 2013 with headaches. Neurologic examination was nonfocal, funduscopy was normal, and there were no myelopathic signs. Interval increase and worsening of the diffuse pachymeningeal thickening with enhancement involving the posterior fossa and extending into the cervical canal along with mass effect on the inferior aspect of the fourth ventricle with slight protrusion of the cerebellar tonsils through the foramen magnum was documented on MRI. The progressive ventriculomegaly was consistent with hydrocephalus (figures 2 and 3). Cervical spine MRI revealed exaggerated lordosis with diffuse thickening and enhancement of the dura causing mass effect and compression of the cervical cord up to T2-T3 level (figure 4). Her headaches were believed to be congruous with either chronic tonsillar herniation or progressive noncommunicating hydrocephalus. While endoscopic third ventriculostomy was entertained as a therapeutic option, since on examination there was no objective evidence of raised intracranial pressure (i.e., papilledema, altered cognition, long tract signs), it was decided to follow her with surveillance imaging and serial ophthalmologic examinations. She is currently asymptomatic on prednisone. Idiopathic hypertrophic pachymeningitis is an uncommon disorder that causes localized or diffuse thickening of the cranial and spinal dura mater. Untreated, the clinical course is marked by severe headaches, progressive neurologic deterioration, and vision loss. Though steroid-responsive, clinical manifestations frequently recur when steroid and immunosuppressive agents are tapered off.1

Intervention Effects of Ganoderma Lucidum Spores on Epileptiform Discharge Hippocampal Neurons and Expression of Neurotrophin-4 and N-Cadherin

Epilepsy can cause cerebral transient dysfunctions. Ganoderma lucidum spores (GLS), a traditional Chinese medicinal herb, has shown some antiepileptic effects in our previous studies. This was the first study of the effects of GLS on cultured primary hippocampal neurons, treated with Mg2+ free medium. This in vitro model of epileptiform discharge hippocampal neurons allowed us to investigate the anti-epileptic effects and mechanism of GLS activity. Primary hippocampal neurons from <1 day old rats were cultured and their morphologies observed under fluorescence microscope. Neurons were confirmed by immunofluorescent staining of neuron specific enolase (NSE). Sterile method for GLS generation was investigated and serial dilutions of GLS were used to test the maximum non-toxic concentration of GLS on hippocampal neurons. The optimized concentration of GLS of 0.122 mg/ml was identified and used for subsequent analysis. Using the in vitro model, hippocampal neurons were divided into 4 groups for subsequent treatment i) control, ii) model (incubated with Mg2+ free medium for 3 hours), iii) GLS group I (incubated with Mg2+ free medium containing GLS for 3 hours and replaced with normal medium and incubated for 6 hours) and iv) GLS group II (neurons incubated with Mg2+ free medium for 3 hours then replaced with a normal medium containing GLS for 6 hours). Neurotrophin-4 and N-Cadherin protein expression were detected using Western blot. The results showed that the number of normal hippocampal neurons increased and the morphologies of hippocampal neurons were well preserved after GLS treatment. Furthermore, the expression of neurotrophin-4 was significantly increased while the expression of N-Cadherin was decreased in the GLS treated group compared with the model group. This data indicates that GLS may protect hippocampal neurons by promoting neurotrophin-4 expression and inhibiting N-Cadherin expression.

Influence of vagal injury on acute traumatic reaction after blast injury.

A prospectively randomised, controlled animal study was conducted to analyse the influence of vagal injury on acute cardio-respiratory responses to blast injury. We used a previously described model of blast-fragment combined injuries to divide dogs randomly into three groups: normal control, blast injury with fragment shot at the masseter and blast injury with fragment shot at the neck. The vagal histomorphologic changes were investigated by haematoxylin-eosin staining and immunocytochemical analysis of neuron-specific enolase and glial fibrillary acidic protein. The indices of respiration, heart rate, blood pressure and body temperature were recorded continuously before and after blast exposure. The vagal injury was more severe in the neck-injured than in the face-injured group. However, bradycardia, hypotension and absence of compensatory peripheral vasoconstriction, which are typically seen in animals subjected to blast injury without vagal nerve injuries, were partly inhibited when the vagal nerve was injured. A vagally mediated reflex, such as a cardio-respiratory system defensive reflex that caused shock, was observed immediately after blast pressure wave injury. These observations may have important implications for the emergency management of blast injury cases.

Central granular cell odontogenic tumor: The first reported case in Oriental people and literature review

The central granular cell odontogenic tumor (CGCOT) is a rare odontogenic neoplasm, usually occurring in the mandible of middle-aged women. Previous studies have reported only 34 cases, all of whom were white or black individuals. The present study reports an additional case of CGCOT, occurring in the posterior mandible of a 69-year-old Taiwanese man. To the authors' knowledge, this is the first reported case in Oriental people. The odontogenic epithelium exhibited strong positive immunoreactivity for pan-cytokeratin, and focal weak staining for bcl-2. The granular cells showed strong positivity for vimentin and α1-antichymotrypsin, and focal weak staining for carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and CD68. These features indicated a mesenchymal origin and possible histiocytic lineage for the granular cells. This study also presents a literature review and describes immunohistochemical features of the tumor.