Vagal nerve and the gastric mucosal defense

An essential role for an intact vagal nerve has been proven in the development of gastric mucosal cyto- and general protection. On the other hand, chemically-induced (ethanol, HCl, indomethacin) gastric mucosal damage is enhanced after acute surgical vagotomy. The aims of this paper were to study the possible mechanisms of the vagal nerve in the development of gastric mucosal defense. The following questions were addressed: 1) effect of surgical vagotomy on the development of ethanol- (ETOH), HCl-, and indomethacin (IND)-induced gastric mucosal damage; 2) changes in the gastric mucosal defense by scavengers, prostacyclin and other compounds (small doses of atropine and cimetidine: 3) changes in the gastric mucosal vascular permeability due to chemicals; 4) effect of indomethacin in the ETOH and HCl models with and without surgical vagotomy; 5) changes in the gastric mucosal content of prostacyclin and PGE₂ in the ETOH and HCl models after surgical vagotomy; and 6) changes in the role of SH-groups in gastric mucosal defense after surgical vagotomy. It was found that: 1) the gastric mucosal damage produced by chemicals (ETOH, HCl, and indomethacin) was enhanced after surgical vagotomy; 2) the cyto- and general gastric protective effects of β-carotene, prostacyclin, and small doses of atropine and cimetidine disappeared after surgical vagotomy; 3) the vascular permeability due to chemicals (ETOH, HCl, indomethacin) significantly increased after surgical vagotomy in association with an increase in both number and severity of gastric mucosal lesions; 4) IND alone (in animals with an intact vagus) did not produce gastric mucosal lesions (in 1-h experiments), but it aggravated ETOH-induced gastric mucosal damage (both its number and severity); 5) the gastric mucosal levels of prostacyclin and PGE₂ decreased after surgical vagotomy; 6) IND application (after surgical vagotomy) decreased further the tissue levels of prostacyclin and PGE₂ in association with an increase of gastric mucosal damage; and 7) the gastric mucosal protective effects of SH-groups were abolished by surgical vagotomy.     

Dampening HOTAIR sensitizes the gastric cancer cells to oxaliplatin through miR-195-5p and ABCG2 pathway

Long non-coding RNAs (lncRNA) have an extensive role in the progression and chemoresistance of gastric cancer (GC). Deeply study the regulatory role of lncRNAs could provide potential therapeutic targets. The aim of this study is to explore the regulatory role of HOTAIR in the progression and oxaliplatin resistance of GC. The expression of HOTAIR in GC and cell lines were detected by using qRT-PCR. Cell proliferation and apoptosis were analysed by CCK-8, EdU incorporation and flow cytometry. Luciferase reporter assay was used to identify the interaction between HOTAIR and ABCG2 (ATP-binding cassette (ABC) superfamily G member 2, ABCG2) via miR-195-5p. The regulatory functions were verified by using molecular biology experiments. HOTAIR was significantly overexpressed in GC and associated with poor prognosis. Knock-down of HOTAIR inhibited the GC cells proliferation and oxaliplatin resistance, while overexpression of HOTAIR showed opposite functions. Further studies found that HOTAIR acted as a competing endogenous RNA (ceRNA) to absorb miR-195-5p and elevated the expression of ABCG2, which leads to resistance of GC cells to oxaliplatin. Taken together, our findings demonstrated that HOTAIR regulates ABCG2 induced resistance of GC to oxaliplatin through miR-195-5p signalling and illustrate the great potential of developing new therapeutic targets for GC patients.     

血清TAP、TFF3与晚期胃癌患者含奥沙利铂化疗方案敏感性和预后的关系

摘要 目的：探讨血清肿瘤异常蛋白（TAP）、三叶因子3（TFF3）与晚期胃癌患者应用含奥沙利铂化疗方案敏感性和预后的关系。方法：选择2017年1月至2020年1月河北大学附属医院收治的115例晚期胃癌患者,所有患者接受含奥沙利铂化疗方案治疗,根据疗效分为敏感组（47例）和耐药组（68例）。化疗前检测血清TAP、TFF3水平,受试者工作特征（ROC）曲线分析TAP、TFF3预测晚期胃癌患者接受含奥沙利铂化疗疗效的价值。治疗后随访,Wilcoxon检验不同血清TAP、TFF3表达下晚期胃癌患者中位OS时间差异。结果：耐药组血清TAP、TFF3水平高于敏感组（P＜0.05）。TAP、TFF3预测晚期胃癌患者含奥沙利铂化疗耐药的曲线下面积分别为0.717、0.690,联合TAP和TFF3预测晚期胃癌患者含奥沙利铂化疗耐药的曲线下面积为0.801,高于单独TAP、TFF3单独检测。随访期间失访2例,死亡54例,高水平TAP、高水平TFF3晚期胃癌患者中位OS时间短于低水平TAP、低水平TFF3晚期胃癌患者（P＜0.05）。结论：对含奥沙利铂化疗耐药的晚期胃癌患者血清TAP、TFF3水平显著增高,高水平TAP、TFF3晚期胃癌患者中位OS时间较短,联合检测血清TAP和TFF3可预测晚期胃癌患者化疗反应性和预后。     

Heat shock in mechanically wounded carrot root disks causes destabilization of stable secretory protein mRNA and dissociation of endoplasmic reticulum lamellae

In many organisms, the synthesis of heat shock proteins during heat shock is concomitant with the cessation of at least a portion of normal cellular protein synthesis. Heat shocked barley aleurone layers selectively stop the synthesis and secretion of secretory proteins. Exposure to 40°C causes a disruption of endoplasmic reticulum (ER) lamellae, which we have hypothesized leads to the destabilization of otherwise stable mRNA previously associated with ER‐bound polyribosomes. We report here that this was also observed in wounded carrot ( Daucus carota L.) root parenchyma tissue which synthesizes and secretes cell wall proteins when mechanically wounded. Nondenaturing cationic polyacrylamide gel electrophoresis of radiolabeled proteins indicated that heat shock caused the cessation of the synthesis and secretion of extensin, a hydroxyproline‐rich cell wall glycoprotein. Northern blot analyses indicated that the mRNA levels for both extensin and another cell wall protein (p33) were rapidly diminished during heat shock. Under nonheat shock conditions extensin mRNA had a half‐life of greater than 4 h, but this appeared to be reduced to less than 30 min during heat shock. There was also a concomitant dissociation of ER lamellae in wounded, heat shocked carrot root tissue, as observed by transmission electron microscopy. These observations indicate that this response may be universal among plant secretory tissues.     

Chemico-structure of the organophosphatic shells of siphonotretide brachiopods

The organophosphatic shell of siphonotretide brachiopods is stratiform with orthodoxly secreted primary and secondary layers. The dominant apatitic constituents of the secondary layer are prismatic laths and rods arranged in monolayers (occasionally in cross-bladed successions), normally recrystallized as platy laminae. Sporadically distributed, interlaminar, lenticular chambers, containing apatitic meshes of laths and aggregates of plates and spherulites, probably represent degraded, localized exudations of glycosaminoglycans (GAGs) with dispersed apatite.
The shells of Helmersenia and Gorchakovia are perforated by canals with external depressions (antechambers) that possibly contained chitinous tubercles in vivo . The immature shell of Siphonotreta and most other siphonotretids is similarly perforated and pitted; but the mature part bears recumbent, rheomorphic, hollow spines that grew forward out of pits. Internally, spines pierce the shell as independent structures to terminate as pillars in GAGs chambers. Spines and pillars were probably secreted by collectives of specialized cells (acanthoblasts) within the mantle.
The shell of the oldest siphonotretide, Schizambon , is imperforate but the ventral valve has a pedicle foramen that lies forward of the posterior margin of the juvenile valve. This relationship characterizes all siphonotretides, suggesting that the pedicle, in vivo , originated within the ventral outer epithelium and not from the posterior body wall as in lingulides.     

Salt gland function in the common eider duck (Somateria mollissima)

The function of the supra-orbital salt gland was studied in the common eider duck (Somateria mollissima). The maximum salt-secreting capacity was determined in (1) wild ducks which had been living in a marine environment, (2) ducks reared in captivity on fresh water, and (3) ducks from group 2 adapted to salt water. The maximum secreting capacity was found by infusing a solution of NaCl (1000 mosmol·kg^-1) at increasing rates, from 0.691 to 1.671 mosmol·min^-1. Freshwater-adapted ducks secreted at a maximum rate of 0.785 mosmol·min^-1 (1500 mosmol·kg^-1). Adapted to salt water they increased their capacity, and the best duck secreted at a rate of 1215 mosmol·min^-1 (1600 mosmol·kg^-1). The best wild duck secreted at a rate of 1516 mosmol·min^-1. Ducks in group 3 were used to examine the response to a hyperosmotic or an isoosmotic infusion. The amount of salt (NaCl) given per unit time was the same. Given a hyperosmotic solution their salt glands secreted at a high rate: 30 min after the infusion had stopped the ducks had excreted 94% of the sodium infused, 92.9% via the salt gland. Given an isoosmotic solution they secreted at a rate about half the infusion rate: 30 min after cessation of infusion they had excreted 73% of the sodium, 42.9% via the salt gland and the rest by the kidneys.Abbreviations A II angiotensin II - AV I arginine vasotocin - ED freshwater-adapted ducks - FW fresh water - SD saltwater-adapted ducks - SW sea water - WD wild ducks     

Effects of peptide YY and its analogues on chloride ion secretion in fed and fasted rat jejunum

The aim of our study was to determine whether a meal modifies the antisecretory response induced by PYY and the structural requirements to elicit antisecretory effects of analogue PYY(22–36) for potential antidiarrhea therapy. The variations in short-circuit current (Isc) due to the modification of ionic transport across the rat intestine were assessed in vitro, using Ussing chambers. In fasted rats, PYY induced a dose- and time-dependent reduction in Isc, with a sensitivity threshold at 5 × 10⁻¹¹ M (ΔIsc −2 ± 0.5 μA/cm²). The reduction was maximal at 10⁻⁷ M (Isc −23 ± 2 μA/cm²), and the concentration producing half-maximal inhibition was 10⁻⁹ M. At 10⁻⁷ M, reduction of Isc by PYY reached 90% of response to 5 × 10⁻⁵ M bumetanide. The PYY effect was partly reversed by 10⁻⁵ M forskolin (Isc +13.43 ± 2.91 μA/h·cm², p < 0.05) or 10⁻³ M dibutyryl adenosine 3′,5′ cyclic monophosphate (Isc +12 ± 1.69 μA/cm², p < 0.05). Naloxone and tetrodotoxin did not alter the effect of PYY. In addition, PYY and its analogue P915 reduced net chloride ion secretion to 2.85 and 2.29 μEq/cm² (p < 0.05), respectively. The antisecretory effect of PYY was accompanied by dose- and time-dependent desensitization when jejunum was prestimulated by a lower dose of peptide. The antisecretory potencies exhibited by PYY analogues required both a C-terminal fragment (22–36) and an aromatic amino acid residue (Trp or Phe) at position 27. At 10⁻⁷ M the biological activity of PYY was lower in fed than fasted rats (p < 0.001). Our results confirm the antisecretory effect of PYY, but show that the fed period is accompanied by desensitization, similar to the transient desensitization observed in the fasted period with cumulative doses. This suggests that PYY may act as a physiological mediator that reduces intestinal secretion.     

Immortalized hypothalamic luteinizing hormone-releasing hormone (LHRH) neurons: A new tool for dissecting the molecular and cellular basis of LHRH physiology

Summary 1. Two LHRH neuronal cell lines were developed by targeted tumorigenesis of LHRH neuronsin vivo. These cell lines (GN and GT-1 cells) represent a homogeneous population of neurons. GT-1 cells have been further subcloned to produce the GT1-1, GT1-3, and GT1-7 cell lines. While considerable information is accumulating about GT-1 cells, very little is currently known about the characteristics and responses of GN cells.2. By both morphological and biochemical criteria, GT-1 cells are clearly neurons. All GT-1 cells immunostain for LHRH and the levels of prohormone, peptide intermediates, and LHRH in the cells and medium are relatively high.3. GT-1 cells biosynthesize, process, and secrete LHRH. Processing of pro-LHRH appears to be very similar to that reported for LHRH neuronsin vivo. At least four enzymes may be involved in processing the prohormone to LHRH.4. LHRH neurons are unique among the neurons of the central nervous system because they arise from the olfactory placode and grow back into the preoptic-anterior hypothalamic region of the brain. Once these neurons reach this location, they send their axons to the median eminence. With respect to the immortalized neurons, GN cells were arrested during their transit to the brain. In contrast, GT-1 cells were able to migrate to the preoptic-anterior hypothalamic region but were unable correctly to target their axons to the median eminence. These problems in migration and targeting appear to be due to expression of the simian virus T-antigen.5. While GT-1 cells are a homogeneous population of neurons, they are amenable to coculture with other types of cells. Coculture experiments currently under way should help not only to reveal some of the molecular and cellular cues that are important for neuronal migration and axonal targeting, but they should also highlight the nature of the cellular interactions which normally occurin situ.6. GT-1 cells spontaneously secrete LHRH in a pusatile manner. The interpulse interval for LHRH from these cells is almost identical to that reported for release of LH and LHRHin vivo. GT-1 cells are interconnected by both gap junctions and synapses. The coordination and synchronization of secretion from these cells could occur through these interconnections, by feedback from LHRH itself, and/or by several different compounds that are secreted by these cells. One such compound is nitric oxide.7. GT-1 cells have Na⁺, K⁺, Ca²⁺, and Cl^– channels. Polymerase chain reaction experiments coupled with Southern blotting and electrophysiological recordings reveal that GT-1 cells contain at least five types of Ca²⁺ channels. R-type Ca²⁺ channels appear to be the most common type of channel and this channel is activated by phorbol esters in the GT-1 cells.8. LHRH is secreted from GT-1 cells in response to norepinephrine, dopamine, histamine, GABA (GABA-A agonists), glutamate, nitric oxide, neuropeptide Y, endothelin, prostaglandin E₂, and activin A. Phorbol esters are very potent stimulators of LHRH secretion. Inhibition of LHRH release occurs in response to LHRH, GABA (GABA-B agonists), prolactin, and glucocorticoids.9. Compared to secretion studies, far fewer agents have been tested for their effects on gene expression. All of the agents which have been tested so far have been found either to repress LHRH gene expression or to have no effect. The agents which have been reported to repress LHRH steady-state mRNA levels include LHRH, prolactin, glucocorticoids, nitric oxide, and phorbol esters. While forskolin stimulates LHRH secretion, it does not appear to have any effect on LHRH mRNA levels.     

Characterization of Syrian hamster gastric mucosal H+,K+-ATPase

Employing a simple one-step sucrose gradient fractionation method, gastric mucosal membrane of Syrian hamster was prepared and demonstrated to be specifically enriched in H⁺,K⁺-ATPase activity. The preparation is practically devoid of other ATP hydrolyzing activity and contains high K⁺-stimulated ATPase, activity of at least 4–5 fold compared to basal ATPase activity. The H⁺,K⁺-ATPase showed hydroxylamine-sensitive phosphorylation and K⁺-dependent dephosphorylation of the phospho-enzyme, characteristic inhibition by vanadate, omeprazole and SCH 28080, and nigericin-reversible K⁺-dependent H⁺-transport — properties characteristic of gastric proton pump One notable difference with H⁺,K⁺-ATPase of other species has been the observation of valinomycin-independent H⁺ transport in such membrane vesicles. It is proposed that such H⁺,K⁺-ATPase-rich hamster gastric mucosal membrane preparation might provide a unique model to study physiological aspects of H⁺,K⁺-ATPase-function in relation to HCl secretion.     

Angiotensin II Receptor Type I-Regulated Anion Secretion in Cystic Fibrosis Pancreatic Duct Cells

The β-adrenergic (cAMP-dependent) regulation of Cl⁻ conductance is defective in cystic fibrosis (CF). The present study explored alternative regulation of anion secretion in CF pancreatic ductal cells (CFPAC-1) by angiotensin II (AII) using the short-circuit current (I _SC ) technique. An increase in I _SC could be induced in CFPAC-1 cells by basolateral or apical application of AII in a concentration-dependent manner (EC₅₀ at 3 μm and 100 nm, respectively). Angiotensin receptor subtypes were identified using specific antagonists, losartan and PD123177, for AT₁ and AT₂ receptors, respectively. It was found that losartan (1 μm) could completely inhibit the AII-induced I _SC , whereas, PD123177 exerted insignificant effect on the I _SC , indicating predominant involvement of AT₁ receptors. The presence of AT₁ receptors in CFPAC-1 cells was also demonstrated by immunohistochemical studies using specific antibodies against AT₁ receptors. Confocal microscopic study demonstrated a rise in intracellular Ca²⁺ upon stimulation by AII indicating a role of intracellular Ca²⁺ in mediating the AII response. Depletion of intracellular but not extracellular pool of Ca²⁺ diminished the AII-induced I _SC . Treatment of the monolayers with a Cl⁻ channel blocker, DIDS, markedly reduced the I _SC , indicating that a large portion of the AII-activated I _SC was Cl⁻-dependent. AII-induced I _SC was also observed in monolayers whose basolateral membranes had been permeabilized by nystatin, suggesting that the I _SC was mediated by apical Cl⁻ channels. Our study indicates an AT₁-mediated Ca²⁺-dependent regulatory mechanism for anion secretion in CF pancreatic duct cells which may be important for the physiology and pathophysiology of the pancreas. Received: 17 June 1996/Revised: 14 November 1996