褶纹冠蚌脏神经节Glu、GABA、DA、Ach和5-HT能神经元的分布

运用H.E.染色法和SP免疫组织化学技术,观察了褶纹冠蚌(Cristaria plicata(Leach)中枢神经系统脏神经节的谷氨酸(glutamate,Glu)、γ-氨基丁酸(γ-aminobutyric acid,GABA)、多巴胺(dopamine,DA)、乙酰胆碱(acetylcholine,Ach)和5-羟色胺(5-hydroxytryptamine,5-HT)神经元的分布和形态。Glu神经元、GABA神经元和DA神经元多为小型细胞,Ach神经元多为中型细胞,5-HT神经元多为大、中型细胞,偶见小型细胞。并且DA神经元分布在脏神经节的整个胞体区,Ach神经元多分布于近外套神经和鳃神经根部,而在中央纤维网区域存在大量GABA能神经纤维。结果表明,褶纹冠蚌脏神经节内含有Glu、GABA、DA、Ach和5-HT能神经元,但这5种神经元的形态和分布有所不同。

海湾扇贝促性腺激素释放激素基因克隆及表达分析

促性腺激素释放激素(Gonadotropin Releasing Hormone,GnRH)是脊椎动物下丘脑-垂体-性腺轴的关键信号分子。近年来的研究显示,GnRH也存在于贝类等无脊椎动物,且在雌雄异体性别分化与繁殖中起重要作用,然而雌雄同体贝类GnRH的研究仍处于空白。本文以海湾扇贝为研究对象,利用RACE技术克隆获得GnRH的cDNA全长序列,该基因长1335 bp,编码101个氨基酸,前体包含1个由24个氨基酸组成的信号肽以及1个具有酰胺化修饰的活性肽(QNFHYSNGWQP-amide)。GnRH在脑足神经节和脏神经节中均有表达,且在2015年10月-2016年1月,GnRH在脑足神经节中的表达量显著高于脏神经节(P<0.05);GnRH在两个神经节中的表达均呈现先上升后下降的趋势,峰值出现在11月份,推测可能与性腺发育启动有关。本研究为深入理解GnRH在贝类繁殖调控中的作用奠定了基础。

Effect of NCPB and VSPL on pain and quality of life in chronic pancreatitis patients

AIM: To compare the effects of neurolytic celiac plexus block (NCPB) and videothoracoscopic splanchnicectomy (VSPL) on pain and quality of life of chronic pancreatitis (CP) patients.METHODS: Forty-eight small duct CP patients were treated invasively with NCPB (n = 30) or VSPL (n = 18) in two non-randomized, prospective, case-controlled protocols due to chronic pain syndrome, and compared to a control group who were treated conservatively (n = 32). Visual analog scales were used to assess pain and opioid consumption rate was evaluated. In addition, the quality of life was measured using QLQ C-30 for NCPB and FACIT for VSPL.Although both questionnaires covered similar problems,they could not be compared directly one with another.Therefore, the studies were compared by meta-analysis methodology.RESULTS: Both procedures resulted in a significant positive effect on pain of CP patients. Opioids were withdrawn totally in 47.0% of NCPB and 36.4% of VSPL patients,and reduced in 53.0% and 45.4% of the respective patient groups. No reduction in opioid usage was observed in the control group. In addition, fatigue and emotional well-being showed improvements. Finally, NCPB demonstrated stronger positive effects on social support, which might possibly be attributed to earlier presentation of patients treated with NCPB.CONCLUSION: Both invasive pain treatment methods are effective in CP patients with chronic pain.

Hepatic schwannoma:Imaging findings on CT,MRI and contrast-enhanced ultrasonography

A primary benign schwannoma of the liver is extremely rare and is difficult to preoperatively discriminate from a malignant tumor.We compared the imaging and pathological findings,and examined the possibility of preoperatively diagnosing a benign liver schwannoma.A 72-year-old woman was admitted to our hospital because of a 4.6-cm mass in the liver.A malignant tumor was suspected,and a right hepatectomy was performed.After this,the diagnosis of a primary benign schwannoma of the liver was made through pathological examination.Contrast-enhanced ultrasonography(CEUS) with Sonazoid showed minute blood flows into the septum and solid areas of the tumor in the vascular phase;most likely due to increased arterial flow associated with infiltration of chronic inflammatory cells.In the postvascular phase,CEUS showed contrast defect of cystic areas and delayed enhancement of solid areas;most likely due to aggregation of siderophores.Because discriminating between a benign and malignant schwannoma of the liver is difficult,surgery is generally recommended.However,the two key findings from CEUS may be useful in discriminating ancient schwannoma by recognizing the hemorrhage involved in the secondary degeneration and aggregation of siderophores.

Norepinephrine transporter (NET) is expressed in cardiac sympathetic ganglia of adult rat

The sympathetic nervous system plays a cardinal role in regulating cardiac function through releasing the neurotransmitter norepinephrine (NE). In comparison with central nervous system, the molecular mechanism of NE uptake in myocardium is not clear. In present study, we proved that in rat the CNS type of NE transporter (NET) was also expressed in middle cervical-stellate ganglion complex (MC-SG complex) which is considered to control the activity of heart, but not expressed in myocardium. The results also showed that NET expression level in right ganglion was significantly higher than in the left, rendering the greater capacity of NE uptake in right ventricle, a fact which may contribute to the maintenance of right ventricular function under pathologic state.

Electrophysiological evidence of P2X2 receptor expression in the neurons of intracardiac and paratracheal ganglia

Objective： To investigate the expression of P2X receptors on rat intracardiac and paratracheal ganglion neurons. Methods： For preparation of intracardiac neurons, hearts were excised, the atria were separated and the medial region containing intracardiac ganglia was isolated and cut into pieces. For preparation of paratracheal neurons, the tracheas were removed and the superficial membranous layer containing paratracheal ganglia was rapidly isolated. Intracardiac and paratracheal ganglion neurons were dissociated after digestion by collagenase and trypsin. Whole-cell patch clamp recording was used to identify the pharmacological properties of P2X receptors in cultured neurons. Results：Neurons from these two ganglia responded to ATP with a rapidly activating, sustained inward current, αβ-meATP failed to evoke any re- sponses in paratracheal ganglion neurons while a few of intracardiac ganglion neurons responded to αβ- meATP with a tiny sustained inward current. ADP and UTP had no effect on intracardiac neurons. Lowering pH potentiated ATP responses in neurons from these two ganglia whereas increasing pH inhibited ATP responses. Co-application of Zn^2＋ potentiated ATP responses in intracardiac and paratracheal ganglion neurons. Conclusion： The receptor subtypes involved in intracardiac and paratracheal ganglia appear to be homomeric P2X2, while heteromeric P2X2/3 could not be completely excluded from intracardiac neurons.

Acupuncture inhibition on neuronal activity of spinal dorsal horn induced by noxious colorectal distention in rat

AIM: To observe how acupuncture stimulation influences the visceral nociception in rat and to clarify the interactions between acupuncture or somatic input and visceral nociceptive inputs in the spinal dorsal horn. These will provide scientific base for illustrating the mechanism of acupuncture on visceral pain.METHODS: Experiments were performed on SpragueDawley rats and the visceral nociceptive stimulus was generated by colorectal distention (CRD). Unit discharges from individual single neuron were recorded extracellularly with glass-microelectrode in L1-3 spinal dorsal horn.Acupuncture stimulation was applied at contralateral heterotopic acupoint and ipsilateral homotopic acupoint,both of which were innervated by the same segments that innervate also the colorectal-gut.RESULTS: The visceral nociception could be inhibited at the spinal level by the heterotopic somatic mechanical stimulation and acupuncture. The maximal inhibition was induced by acupuncture or the somatic noxious stimulation at spinal dorsal horn level with inhibiting rate of 68.61%and 60.79%, respectively (P＜0.01 and ＜0.001). In reversible spinalized rats (cervical-thoracic cold block)both spontaneous activity and responses to CRD increased significantly in 16/20 units examined, indicating the existence of tonic descending inhibition. The inhibition of acupuncture on the noxious CRD disappeared totally in the reversible spinalized rats (P＜0.001).CONCLUSION: The inputs of noxious CRD and acupuncture may interact at the spinal level. The nociceptive visceral inputs could be inhibited by acupuncture applied to hetero-topic acupoint. The effect indicates that the spinal dorsal horn plays a significant role in mediating the inhibition of acupuncture and somatic stimulation on the neuronal response to the noxious visceral stimulation and the inhibition is modulated by upper cervical cord and/or supra-spinal center.

Effect of bombesin and neurotensin on gut barrier function in partially hepatectomized rats

AIM： To investigate the effect of regulatory peptides bombesin （BBS） and neurotensin （NT） on intestinal barrier function in partially hepatectomized rats. METHODS： Ninety male Wistar rats were randomly divided into five groups： Ⅰ （n = 10）： controls, Ⅱ （n = 20）： sham operated, Ⅲ （n = 20）： partial hepatectomy 70% （PHx）, Ⅳ （n = 20）： PHx＋BBS （30 μg/kg/d）, Ⅴ （n = 20）： PHx＋NT （300 μg/kg/d）. Groups IV and V were treated for 8 days before PHx and 48 h post surgery. At the end of the experiment, on day 10, intestinal barrier function was assessed by measuring endotoxin concentrations in portal and aortic blood. Tissue sections of the terminal ileum were examined histologically and villus density, mucosal thickness, mitotic activity and apoptosis in crypts were assessed. In addition, ileal mucosa was analyzed for DNA and protein content and microbiological analysis was performed in cecal contents. To estimate intestinal oxidative stress, lipid peroxidation was determined on tissue homogenates from terminal ileum. RESULTS： BBS or NT administration significantly reduced portal and systemic endotoxemia observed 48 h after partial hepatectomy. In hepatectomized rats （group Ⅲ）, a trend towards induction of mucosal atrophy was observed, demonstrated by the reduction of villus density, mucosal thickness, protein content and significant reduction of DNA, while these alterations were reversed by regulatory peptides administration. This trophic effect of BBS and NT was accompanied by induction of mitoses above control levels and a significant reduction of apoptosis in intestinal crypts. Intestinal lipid peroxidation was found significantly lower in PHx group and regulatory peptides exerted an antioxidant action, further decreasing this parameter of oxidative stress. The ：bacterial population of E. coli and aerobic Gram （＋） cocci was increased in cecal content of hepatectomized rats, while this parameter was not affected by the administration of BBS or NT. CONCLUSION： Gut regulatory peptides BBS and NT improve intestinal barrier function and reduce endotoxemia in experimental partial hepatectomy. This effect is, at least in part, mediated by their trophic, antiapoptotic, mitogenic, and antioxidant effect on the intestinal epithelium. This observation might be of potential value in patients undergoing liver resection.

Prevention and treatment of hepatic encephalopathy: Focusing on gut microbiota

The gut flora plays an important role in the pathogenesis of the complications of cirrhosis. Hepatic encephalopathy (HE) represents a broad continuum of neuropsychological dysfunction in patients with acute or chronic liver disease and/or porto-systemic shunting of blood flow and it manifests with progressive deterioration of the superior neurological functions. The pathophysiology of this disease is complex, as it involves overproduction and reduced metabolism of various neurotoxins, particularly ammonia. Management of HE is diversified and requires several steps: elimination of precipitating factors, removal of toxins, proper nutritional support, modulation of resident fecal flora and downregulation of systemic and gut-derived inflammation. This review will provide an overview of gut barrier function and the influence of gut-derived factors on HE, focusing on the role of gut microbiota in the pathogenesis of HE and the recent literature findings on its therapeutic manipulation.

Noncoding RNAs and Their Potential Therapeutic Applications in Tissue Engineering

Tissue engineering is a relatively new but rapidly developing field in the medical sciences. Noncoding RNAs(ncRNAs) are functional RNA molecules without a protein-coding function; they can regulate cellular behavior and change the biological milieu of the tissue. The application of ncRNAs in tissue engineering is starting to attract increasing attention as a means of resolving a large number of unmet healthcare needs, although ncRNA-based approaches have not yet entered clinical practice. In-depth research on the regulation and delivery of ncRNAs may improve their application in tissue engineering.The aim of this review is: to outline essential ncRNAs that are related to tissue engineering for the repair and regeneration of nerve, skin, liver, vascular system, and muscle tissue; to discuss their regulation and delivery; and to anticipate their potential therapeutic applications.

Automatic Liver Segmentation Scheme for MRI Images Based on Cellular Neural Networks

Currently, the processing speed of exist-ing autormtic liver segmentation for Magnetic Res-onance Imaging （MRI） images is rehtively slow. An automatic liver segmentation scheme for MRI irmges based on Cellular Neural Networks （CNN） is presented in this paper. It ensures the validity of this scheme and at the same time completes the im-age segmentation faster to accurately calculate the liver volume by using parallel computing in real time. In order to facilitate the CNN irmge process-hag, firstly, three-dimensional liver MRI images should be transformed into binary images; second- ly, an appropriate template parameter of the Global Connectivity Detection CNN （GCD CNN） shall be selected to probe the connectivity of the liver to extract the entire liver; and then the Hole-Filler CNN （HF CNN） are used to repair the entire extracting liver and improve the accuracy of fiver segmentation; final-ly, the liver volume is obtained. Results show that the scheme can ensure the accuracy of the automatic seg-mentation of the liver, and it can also improve the processing speed at the same time. The liver volume calculated is in line with the clinical diagnosis.

New techniques in the tissue diagnosis of gastrointestinal neuromuscular diseases

Gastrointestinal neuromuscular diseases are a clinically heterogeneous group of disorders of children and adults in which symptoms are presumed or proven to arise as a result of neuromuscular (including interstitial cell of Cajal) dysfunction. Common to most of these diseases are symptoms of impaired motor activity which manifest as slowed or obstructed transit with or without evidence of transient or persistent radiological visceral dilatation. A variety of histopathological techniques and allied investigations are being increasingly applied to tissue biopsies from such patients. This review outlines some of the more recent advances in this fi eld, particularly in the most contentious area of small bowel disease manifesting as intestinal pseudo-obstruction.

Preoperative microcoil embolization of the common hepatic artery for pancreatic body cancer

AIM:To evaluate safety and feasibility of microcoil embolization of the common hepatic artery under proper or distal balloon inflation in preoperative preparation for en bloc celiac axis resection for pancreatic body cancer.METHODS:Fifteen patients(11 males,4 females;median age,67 years) with pancreatic body cancer involving the nerve plexus surrounding the celiac artery underwent microcoil embolization.To alter the total hepatic blood flow from superior mesenteric artery(SMA),microcoil embolization of the common hepatic artery(CHA) was conducted in 2 cases under balloon inflation at the proximal end of the CHA and in 13 cases under distal microballoon inflation at the distal end of the CHA.RESULTS:Of the first two cases of microcoil embolization with proximal balloon inflation,the first was successful,but there was microcoil migration to the proper hepatic artery in the second.The migrated microcoil was withdrawn to the CHA by an inflated microballoon catheter.Microcoil embolization was successful in the other 13 cases with distal microballoon inflation,with no microcoil migration.Compact microcoil embolization under distal microballoon inflation created sufficient resistance against the vascular wall to prevent migration.Distal balloon inflation achieved the requisite 1 cm patency at the CHA end for vascular clamping.All patients underwent en bloc celiac axis resection without arterial reconstruction or liver ischemia.CONCLUSION:To impede microcoil migration to the proper hepatic artery during CHA microcoil embolization,distal microballoon inflation is preferable to proximal balloon inflation.

Psychosocial stress and liver disease status

"Psychosocial stress" is an increasingly common concept in the challenging and highly-demanding modern society of today. Organic response to stress implicates two major components of the stress system, namely the hypothalamic-pituitary-adrenal axis and the sympathetic nervous system. Stress is anamnestically reported by patients during the course of disease, usually accompanied by a decline in their overall health status. As the mechanisms involving glucocorticoids and catecholamines have been deciphered, and their actions on immune cell function deeper understood, it has become clear that stress has an impact on hepatic inflam-matory response. An increasing number of articles have approached the link between psychosocial stress and the negative evolution of hepatic diseases. This article reviews a number of studies on both human populations and animal models performed in recent years, all linking stress, mainly of psychosocial nature, and the evolution of three important liver-related pathological entities: viral hepatitis, cirrhosis and hepatocellular carcinoma.

Efficacy and safety of a novel multi-electrode radiofrequency ablation catheter for renal sympathetic denervation in pigs

Objective To investigate the safety and efficacy of a self-developed novel multi-electrode radiofrequency ablation catheter （Spark） for catheter-based renal denervation （RDN）. Methods A total of 14 experimental miniature pigs were randomly divided into four groups （55°＆ 5-watt, 55°＆ 8-watt, 65°＆ 5-watt, and 65° ＆ 8-watt groups）. Spark was used for left and right renal artery radiofrequency ablation. Blood samples collected from renal arteries and veins as well as renal arteriography were performed on all animals before, immediately after, and three months after procedure to evaluate the effects of Spark on the levels of plasma renin, aldosterone, angiotensin I, and angiotensin II as well as the pathological changes of renal arteries. Results One pig died of an anesthetic accident, 13 pigs successfully underwent the bilateral renal artery ablation. Compared with basic measurements, pigs in all the four groups had significantly decreased mean arterial pres- sure after procedure. Histopathological analysis showed that this procedure could result in intimal hyperplasia, significant peripheral sympa- thetic nerve damage in the renal arteries such as inflammatory cell infiltration and fibrosis in perineurium, uneven distribution of nerve fibers, tissue necrosis, severe vacuolization, fTagmented and unclear nucleoli myelin degeneration, sparse axons, and interruption of continuity. In addition, the renal artery radiofrequency ablation could significantly reduce the levels of plasma renin, aldosterone, angiotensin I, and angio- tensin II in pigs. Conclusions The results suggest that this type of multi-electrode catheter-based radiofrequency ablation could effectively remove peripheral renal sympathetic nerves and reduce the activity of systemic renin-angiotensin system in pigs, thus facilitating the control of systemic blood pressure in pigs.

Cardiac Neurons Firing Preceding Cortical Neurons Firing by Variable Time Equivalent to RP before Conscious Act

The signals and the neuronal mechanisms that underlying the behavior, actions, and action-directed goals in man and animals during conscious state are not fully understood, and the neuro-dynamic mechanisms and the source of these neuronal signals are not authenticated. Temporal judgment alone can neither account for neural signaling necessary for emergence of conscious act nor explain RP （Readiness Potential, the accepted neural correlate time needed for the neurons to fire） that precedes the onset of action or the latency time of 0.5 ms that precedes the conscious act found by Libet. Neuronal feedback mechanisms between the heart and the brain seem feasible and logical suggestions to be considered, so clearly, I would suggest that the onset of a conscious-directed goal, conscious action, freewill, intension, and the neural signals and mechanisms that control them may depend upon the interaction between two sources： （1） the brain and （2） the heart. The temporal-cardiac （neural system） interaction has been well established in heart-brain interaction studies by many workers who found that the work of the heart precedes that of the brain in EEG （electroencephalography） findings in conscious stimulation, which may explain and account for RP time and the 0.5 ms latency period of Libet＇s important findings. According to my hypothesis （AlFaki 2009） and views, the temporal neurons in the soma to-sensory cortex will respond to conscious stimulation only after receiving neuronal signals from the cardiac neurons in the neural plexus of the heart; after variable millisecond equivalent to RP or Libet＇s latency period prior to temporal neuronal firinging in response to conscious act, this time is the time needed by cardiac neurons to process and signal information to the brain through feedback mechanism and heart-brain interaction.

EFFECTS OF INTRAVENOUS FENTANYL ON SPONTANEOUS RENAL SYMPATHETIC NERVE ACTIVITY IN NORMAL AND VAGOTOMIZED RABBITS

Objective To investigate the roles of sympathetic and vagus nerves in hypotension and bradycardia induced by fentanyl. Methods Fourteen rabbits were divided into 2 groups: normal and vagotomized rabbits. Rabbits were anesthetized, paralyzed, and artificial ventilated. Right renal sympathetic nerve was exposed and prepared for recording electrical activity. Fentanyl was injected intravenously in incremental doses of 1, 4, 15, 30, and 50 μg/kg at 10 minutes intervals. Results Fentanyl significantly reduced the spontaneous activity of renal sympathetic nerve, mean arterial pressure, and heart rate above a total dose of 20 μg/kg in both normal and vagotomized rabbits. However, normal rabbits spontaneous sympathetic nerve activity and mean arterial pressure were more depressed than vagotomized rabbits at total doses of 50 and 100 μg/kg. There were no significant difference in the reduction of heart rate between normal and vagotomized rabbits. Conclusion Fentanyl induction of bradycardia and hypotension in rabbits is mainly due to depression of sympathetic nerve activity.

Targeting voltage-gated sodium channels for treatment for chronic visceral pain

Voltage-gated sodium channels (VGSCs) play a fundamental role in controlling cellular excitability,and their abnormal activity is related to several pathological processes,including cardiac arrhythmias,epilepsy,neurodegenerative diseases,spasticity and chronic pain.In particular,chronic visceral pain,the central symptom of functional gastrointestinal disorders such as irritable bowel syndrome,is a serious clinical problem that affects a high percentage of the world population.In spite of intense research efforts and after the dedicated decade of pain control and research,there are not many options to treat chronic pain conditions.However,there is a wealth of evidence emerging to give hope that a more refined approach may be achievable.By using electronic databases,available data on structural and functional properties of VGSCs in chronic pain,particularly functional gastrointestinal hypersensitivity,were reviewed.We summarize the involvement and molecular bases of action of VGSCs in the pathophysiology of several organic and functionalgastrointestinal disorders.We also describe the efficacy of VGSC blockers in the treatment of these neurological diseases,and outline future developments that may extend the therapeutic use of compounds that target VGSCs.Overall,clinical and experimental data indicate that isoform-specific blockers of these channels or targeting of their modulators may provide effective and novel approaches for visceral pain therapy.