[Preliminary study on three-dimensional morphological reconstruction method for external nose defect based on three-dimensional face template].

Objective: To provide a new solution for the digital design of nasal prostheses, this study explores the three-dimensional (3D) facial morphology completion method for external nasal defects based on the non-rigid registration process of 3D face template. Methods: A total of 20 male patients with tooth defect and dentition defect who visited the Department of Prosthodontics, Peking University School and Hospital of Stomatology from June to December 2022 were selected, age 18-45 years old. The original 3D facial data of patients were collected, and the 3D facial data of the external nose defect was constructed in Geomagic Wrap 2021 software. Using the structured 3D face template data constructed in the previous research of the research group, the 3D face template was deformed and registered to the 3D facial data of external nose defect (based on the morphology of non-defective area) by non-rigid registration algorithm (MeshMonk program), and the personalized deformed data of the 3D face template was obtained, as the complemented facial 3D data. Based on the defect boundary of the 3D facial data of the external nose defect, the complemented external nose 3D data can be cut out from the complemented facial 3D data. Then the nasofacial angle and nasolabial angle of the complemented facial 3D data and the original 3D facial data was compared and analyzed, the ratio between the nose length and mid-face height, nose width and medial canthal distance of the complemented facial 3D data was measured, the edge fit between the edge curve of the complemented external nose 3D data and the defect edge curve of the 3D facial data of external nose defect was evaluated, and the morphological difference of the nose between the complemented external nose 3D data and the original 3D facial data was analyzed. Results: There was no significant statistically difference (t=-0.23, P=0.823; Z=-1.72, P=0.086) in the nasofacial angle (28.2°±2.9°, 28.4°±3.5° respectively) and nasolabial angle [95.4°(19.2°), 99.9°(9.5°) respectively] between the 20 original 3D facial data and the complemented facial 3D data. The value of the ratio of nose length to mid-face height in the complemented facial 3D data was 0.63±0.03, and the value of the ratio of nose width to medial canthal distance was 1.07±0.08. The curve deviation (root mean square value) between the edge curve of the complemented external nose 3D data and the defect edge curve of the 3D facial data of external nose defect was (0.37±0.09) mm, the maximum deviation was (1.14±0.32) mm, and the proportion of the curve deviation value within±1 mm was (97±3)%. The distance of corresponding nose landmarks between the complemented facial 3D data and the original 3D facial data were respectively, Nasion: [1.52(1.92)] mm; Pronasale: (3.27±1.21) mm; Subnasale: (1.99±1.09) mm; Right Alare: (2.64±1.34) mm; Left Alare: (2.42± 1.38) mm. Conclusions: The method of 3D facial morphology completion of external nose defect proposed in this study has good feasibility. The constructed complemented external nose 3D data has good facial coordination and edge fit, and the morphology is close to the nose morphology of the original 3D facial data.

[Arthroscopic evaluation of tibial plateau fractures combined with meniscal tears and cruciate ligamentous injuries].

Objectives: To investigate the types of meniscal tears and cruciate ligamentous injuries in patients with tibial plateau fracture(TPF) following arthroscopic examination. Methods: The clinical data of 216 patients with TPF who underwent closed reduction and internal fixation (CRIF) from January 2016 to January 2019 at Trauma Emergency center, the Third Hospital of Hebei Medical University were analyzed retrospectively. There were 147 males (147 knees) and 69 females (69 knees),aged 46.3 years (range: 18 to 80 years). All patients underwent closed reduction for the displaced fracture fragment with the use of bidirectional rapid redactor,and minimally invasive percutaneous plate osteosynthesis. Intra-operative arthroscopic examination was performed to exam the stability of meniscus and the continuity of cruciate ligamentous after CRIF. The percentages and types of meniscal tears and cruciate ligamentous injuries were recorded. Results: The overall percentages of meniscal tears associated with TPFs was 48.6%(105/216). The most common pattern of meniscal tears was longitudinal tears, accounting for 43.8% (46/105), and it occurred most frequently in Schatzker type Ⅱ (58.7%, 27/46). Furthermore, the percentage of meniscal complex tears was 17.1% (18/105), occurring most frequently in Schatzker type Ⅴ (9/18). The overall percentage of cruciate ligamentous injuries associated with TPFs was 17.1% (37/216), and the percentages of anterior cruciate ligament (ACL) injuries was 64.9%(24/37), the percentage of posterior cruciate ligament injuries was 35.1%(13/37). Avulsion fracture was the most common pattern in ACL injuries, accounting for 41.7% (13/24), and all occurred in the tibial insertion site. Conclusions: In the present study, the percentages of meniscal tears and ligamentous injuries in TPFs are 48.6% and 17.1%, respectively. The most common types are meniscal longitudinal tears and ACL injury, occurring most frequently in Schatzker type Ⅱ and Ⅳ, respectively. Recognition of concomitant meniscal tears and cruciate ligamentous injuries in TPFs is helpful for trauma physicians to choose the best surgical treatment.

Somatosensory and motor evoked potentials in dogs with chronic severe thoracolumbar spinal cord injury.

Some dogs that become paraplegic after severe spinal cord injury regain ambulation on the pelvic limbs despite permanent loss of pelvic limb sensation, a phenomenon termed 'spinal walking'. Plastic changes in spinal cord circuitry are thought to mediate this form of recovery but the precise circumstances that favor its development are not known. More information on this phenomenon would be helpful because it might be possible to coax more function in chronically paraplegic animals so improving their, and their owners', quality of life. We analysed the correlation of 'spinal walking' and pelvic limb pain sensation with recordings of scalp and spinal somatosensory and transcranial magnetic motor evoked potentials. We prospectively examined 94 paraplegic dogs (including 53 Dachshunds) that had sustained T10 to L3 spinal cord injury (including 78 dogs with acute intervertebral disc herniation) at a median time of 12.0 months from injury. Nine dogs exhibited 'spinal walking' and nine other individuals had intact pelvic limb pain sensation. Of 34 tested, 12 dogs had recordable scalp somatosensory evoked potentials. Fifty-three of 59 tested dogs had recordable spinal somatosensory evoked potentials, but only six had recordable potentials cranial to the lesion. Twenty-two of 94 tested dogs had recordable transcranial magnetic motor evoked potentials in the pelvic limb(s). There was no apparent association between intact evoked potential recording and either spinal walking or intact pain sensation. We conclude that factors other than influence, or lack of influence, of input carried by spinal cord long tracts mediate recovery of spinal walking.

Polymorphisms of TGFB1, TLE4 and MUC22 are associated with childhood asthma in Chinese population.

OBJECTIVE: To investigate whether the genetic variants of TGFB1, TLE4, MUC22 and IKZF3 are associated with the development of asthma in Chinese children. METHODS: 572 adolescent asthma patients and 590 age-matched healthy controls were included in this study. A total of four SNPs were genotyped, including rs2241715 of TGFB1, rs2378383 of TLE4, rs2523924 of MUC22, and rs907092 of IKZF3. Allele frequencies of the patients and the control group were compared by the Chi-square test. The Student t test was used to analyse the relationship between genotypes and clinical feature of the patients. RESULTS: Patients were found to have significantly different frequencies of allele A of rs2241715, allele G of rs2378383 and allele A of rs2523924 as compared with the controls (40.4% vs. 45.9%, p=0.01 for rs2241715; 17.2% vs. 13.4%, p=0.01 for rs2378383; 15.3% vs. 11.9%, p=0.02 for rs2523924). For patients with severe asthma, those with genotype AA/AG of rs2241715 had remarkably higher FEV1% as compared with those with genotype GG (59.1±4.3% vs. 55.4±3.7%, p<0.001). Moreover, those with genotype GG/GA of rs2378383 had remarkably lower FEV1% as compared with those with genotype AA (54.6±2.9% vs. 58.6±4.1%, p<0.001). CONCLUSIONS: Genes TGFB1, TLE4 and MUC22 are associated with the risk of childhood asthma in Chinese population. Our results associating TGFB1 and TLE4 with clinical features of asthma suggest potential application of these parameters in the management of asthma children.

Pathophysiology, Clinical Importance, and Management of Neurogenic Lower Urinary Tract Dysfunction Caused by Suprasacral Spinal Cord Injury.

Management of persistent lower urinary tract dysfunction resulting from severe thoracolumbar spinal cord injury can be challenging. Severe suprasacral spinal cord injury releases the spinal cord segmental micturition reflex from supraspinal modulation and increases nerve growth factor concentration in the bladder wall, lumbosacral spinal cord, and dorsal root ganglion, which subsequently activates hypermechanosensitive C-fiber bladder wall afferents. Hyperexcitability of bladder afferents and detrusor overactivity can cause urine leaking during the storage phase. During urine voiding, the loss of supraspinal control that normally coordinates detrusor contraction with sphincter relaxation can lead to spinal cord segmental reflex-mediated simultaneous detrusor and sphincter contractions or detrusor-sphincter dyssynergia, resulting in inefficient urine voiding and high residual volume. These disease-associated changes can impact on the quality of life and life expectancy of spinal-injured animals. Here, we discuss the pathophysiology and management considerations of lower urinary tract dysfunction as the result of severe, acute, suprasacral spinal cord injury. In addition, drawing from experimental, preclinical, and clinical medicine, we introduce some treatment options for neurogenic lower urinary tract dysfunction that are designed to: (1) prevent urine leakage arising because of detrusor overactivity during bladder filling, (2) preserve upper urinary tract integrity and function by reducing intravesical pressure and subsequent vesicoureteral reflux, and (3) prevent urinary tract and systemic complications by treating and preventing urinary tract infections.

LncRNA UCA1 promotes epithelial-mesenchymal transition (EMT) of breast cancer cells via enhancing Wnt/beta-catenin signaling pathway.

OBJECTIVE: LncRNA UCA1 can promote invasion of breast cancer cells. However, the underlying mechanism is not quite clear. In this study, we investigated the regulative effect of UCA1 on the invasion capability of breast cancer cells and its association with the Wnt/ß-catenin pathway. MATERIALS AND METHODS: Human breast cancer cell line MDA-MB-231 cells were transfected for UCA1 knockdown using UCA1 si-RNA. Transwell assay was performed to assess cell invasion capability. Western blot analysis was conducted to investigate the expression of mesenchymal and epithelial markers and the proteins involved in Wnt/beta-catenin signaling pathway. Immunofluorescent staining was further performed to verify the expression of E-cadherin and N-cadherin. RESULTS: MDA-MB-231 cells have strong invasion capability. Knockdown of endogenous UCA1 significantly reduced the number of invading cells. MDA-MB-231 cells with UCA1 knockdown had significantly increased expression of E-cadherin but decreased expression of N-cadherin, Vimentin and Snail. UCA1 inhibition substantially increased the expression of p-GSK-3ß and GSK-3ß and suppressed the protein expression of ß-catenin and transcription of the downstream genes, including cyclin D1 and MMP-7. CONCLUSIONS: UCA1 can modulate epithelial-mesenchymal transition (EMT) of MDA-MB-231 cells and knockdown of UCA1 impaired the mesenchymal properties. UCA1 upregulation increases invasiveness of breast cancer cells at least partly via activating the Wnt/ß-catenin signaling pathway.

Histopathological investigation of syringomyelia in the Cavalier King Charles spaniel.

Syringomyelia (SM) in Cavalier King Charles spaniels (CKCSs) is identified commonly on magnetic resonance images and is sometimes associated with clinical signs of pain and cervical hyperaesthesia. However, the mechanism by which SM develops in this breed has not been fully elucidated and the associated effects on spinal cord structure have not been reported previously. The aims of this study were to describe changes found in the spinal cord of CKCSs, to compare findings between symptomatic and asymptomatic dogs and to determine whether syrinx formation was associated with tissue destruction. Anomalies of the central canal were found in all specimens and many dogs had grossly visible fluid-filled cavities within the spinal cord. Prominent microscopical findings were spongy degenerative changes associated with neuronal necrosis and Wallerian degeneration. The ependyma was discontinuous in many specimens, notably in symptomatic individuals, and there was evidence of angiogenesis and fibrous tissue proliferation around blood vessels adjacent to syrinx cavities. Compared with two different samples of the normal dog population, dogs with syrinxes had significantly less grey matter, although this decrease was associated with generalized loss of spinal cord area. Therefore, SM is associated with degenerative changes in the spinal cord and may develop through primary disruption of ependymal integrity followed by vascular hypertrophy and proliferation. Glial and fibrous proliferation appears to be associated with expression of clinical signs.

Variation of human papillomavirus 16 in cervical and lung cancers in Sichuan, China.

Although the crucial role of human papillomaviruses (HPVs), especially HPV-16 in various cancers has been confirmed, the variation of HPV-16 among different cancers have not been investigated in a specific geographic location. In order to elucidate whether similar HPV-16 variants are involved in different kinds of cancers in the same geographic location, the analysis of sequence variants of E6 and E7 oncogenes and L1 gene of HPV-16 in cervical and lung cancers in Sichuan, China, was carried out. Tissue samples from 122 cervical cancers, 104 lung cancers, and 138 controls were subjected to RT-PCR or PCR, sequencing, and sequence analysis. The infection rates of HPV-16 in cervical, lung cancers, and non-malignant controls were 68.9%, 17.3%, and 37.0%, respectively. Asian prototype variants prevailed in cervical and lung cancers, while European prototype variants in non-malignant controls. In comparison to the lung cancer, cervical cancer showed a much higher diversity of HPV-16 oncogenes. These results indicate that in Sichuan, China, Asian prototype variants of HPV-16 are more pathogenic than their European counterparts.

Unaltered graft survival and intragraft lymphocytes infiltration in the cardiac allograft of Cxcr3-/- mouse recipients.

Previous studies showed that absence of chemokine receptor Cxcr3 or its blockade prolong mouse cardiac allograft survival. We evaluated the effect of the CXCR3 receptor antagonist MRL-957 on cardiac allograft survival, and also examined the impact of anti-CXCR3 mAb in human CXCR3 knock-in mice. We found only a moderate increase in graft survival (10.5 and 16.6 days, p < 0.05) using either the antagonist or the antibody, respectively, compared to control (8.7 days). We re-evaluated cardiac allograft survival with two different lines of Cxcr3(-/-) mice. Interestingly, in our hands, neither of the independently derived Cxcr3(-/-) lines showed remarkable prolongation, with mean graft survival of 9.5 and 10.8 days, respectively. There was no difference in the number of infiltrating mononuclear cells, expansion of splenic T cells or IFN-gamma production of alloreactive T cells. Mechanistically, an increased other chemokine receptor fraction in the graft infiltrating CD8 T cells in Cxcr3(-/-) recipients compared to wild-type recipients suggested compensatory T-cell trafficking in the absence of Cxcr3. We conclude Cxcr3 may contribute to, but does not govern, leukocyte trafficking in this transplant model.

Neurogastroenterology of tegaserod (HTF 919) in the submucosal division of the guinea-pig and human enteric nervous system.

Actions of the 5-HT(4) serotonergic receptor partial agonist, tegaserod, were investigated on mucosal secretion in the guinea-pig and human small intestine and on electrophysiological behaviour of secretomotor neurons in the guinea-pig small intestinal submucosal plexus. Expression of 5-HT(4) receptor protein and immunohistochemical localization of the 5-HT(4) receptor in the submucosal plexus in relation to expression and localization of choline acetyltransferase and the vesicular acetylcholine (ACh) transporter were determined for the enteric nervous system of human and guinea-pig small intestine. Immunoreactivity for the 5-HT(4) receptor was expressed as ring-like fluorescence surrounding the perimeter of the neuronal cell bodies and co-localized with the vesicular ACh transporter. Exposure of mucosal/submucosal preparations to tegaserod in Ussing chambers evoked increases in mucosal secretion reflected by stimulation of short-circuit current. Stimulation of secretion had a relative high EC(50) of 28.1 +/- 1.3 mumol L(-1), was resistant to neural blockade and appeared to be a direct action on the secretory epithelium. Tegaserod acted at presynaptic 5-HT(4) receptors to facilitate the release of ACh at nicotinic synapses on secretomotor neurons in the submucosal plexus. The 5-HT(2B) receptor subtype was not involved in actions at nicotinic synapses or stimulation of secretion.

The P2Y purinergic receptor expressed by enteric neurones in guinea-pig intestine.

Electrophysiological recording methods provided evidence for presynaptic release of ATP from enteric neurones and postganglionic sympathetic fibres in the enteric nervous system (ENS) of guinea-pig intestine (J Physiol Lond 2003; 550: 493-504). The released ATP acted at postsynaptic P2Y(1) receptors to evoke slow synaptic excitation in neurones in the submucosal division of the ENS. Here, we report the cloning and characterization of the P2Y(1) receptor, which was found in the guinea-pig submucosal layer. A 1178 bp cDNA clone was isolated from guinea-pig submucosal RNA by reverse transcription polymerase chain reaction (RT-PCR). The cDNA contained an open-reading frame of 1119 bp, encoding a 373 amino acid polypeptide of the same length and with 95% identity to the human P2Y(1) receptor. Stable expression of the guinea-pig cDNA in human embryonic kidney (HEK)293 cells was accompanied by a marked increase in sensitivity for elevation of free intracellular calcium evoked by ATP or related nucleotides. The potency order for ATP and its analogues was: 2-methio-adenosine diphosphate > 2-methio-adenosine triphosphate > ADP > ATP-gamma-S > ATP. The selective P2Y(1) receptor antagonist, MRS2179, was a competitive antagonist for the receptor with a pA(2) value of 6.5. The results add to existing evidence for expression of a functional P2Y(1) purinergic receptor in neurones of the submucosal division of the ENS.

Delaying acute graft-versus-host disease in mouse bone marrow transplantation by treating donor cells with antibodies directed at l-selectin and alpha4-integrin prior to infusion.

Acute graft-versus-host disease (GVHD) is still a major hurdle for successful bone marrow transplantation (BMT). Although many immunosuppressive drugs are available, none of them alone or in combination are able to completely abolish acute GVHD. The lifelong immunosuppression profoundly reduces the quality of life of BMT recipients. Therefore, new therapeutic approaches are needed. We previously reported that, in an acute GVHD model using SCID mice as recipient, incubating donor spleen cells with antibodies directed at CD49d and CD62L could significantly delay the occurrence of acute GVHD. To test the potential usefulness of this treatment in BMT, we examined this therapeutic protocol in a mouse BMT model. The present mouse BMT study confirmed our previous results that incubation of donor cells with antibodies directed at CD49d and CD62L prior to infusion into the recipient can effectively delay acute GVHD, allowing the recipients to recover from the side effects of total body irradiation. This one-time treatment is easy and simple and may be modified for clinical usage.

Slow excitatory synaptic transmission mediated by P2Y1 receptors in the guinea-pig enteric nervous system.

Electrophysiological recording was used to study a type of slow excitatory postsynaptic potential (slow EPSP) that was mediated by release of ATP and its action at P2Y1 receptors on morphologically identified neurones in the submucosal plexus of guinea-pig small intestine. MRS2179, a selective P2Y1 purinergic receptor antagonist, blocked both the slow EPSP and mimicry of the EPSP by exogenously applied ATP. Increased conductance accounted for the depolarization phase of the EPSP, which occurred exclusively in neurones with S-type electrophysiological behaviour and uniaxonal morphology. The purinergic excitatory input to the submucosal neurones came from neighbouring neurones in the same plexus, from neurones in the myenteric plexus and from sympathetic postganglionic neurones. ATP-mediated EPSPs occurred coincident with fast nicotinic synaptic potentials evoked by the myenteric projections and with noradrenergic IPSPs evoked by sympathetic fibres that innervated the same neurones. The P2Y1 receptor on the neurones was identified as a metabotropic receptor linked to activation of phospholipase C, synthesis of inositol 1,4,5-trisphosphate and mobilization of Ca2+ from intracellular stores.

T cell infiltration and chemokine expression: relevance to the disease localization in murine graft-versus-host disease.

Acute graft-versus-host disease (GVHD) involves mainly skin, liver and intestines. Other organs such as heart, muscle and central nervous system are seldom affected, although their parenchymal cells also express alloantigens, such as MHC class I antigens. The mechanism of this selective involvement of distinct organs in acute GVHD is not well understood. We postulated that it might be related to the selective migration of activated alloreactive T cells. Indeed, T cell infiltration, revealed by examination of serial samples using flow cytometry and immunohistology, occurred early and continuously in the target organs such as the liver, but not in a non-target organ, the heart, in a murine acute GVHD model. Since T cell migration is largely controlled by the expression of chemokine and chemokine receptors, we investigated the chemokine spectrum in target/non-target organs of mice with acute GVHD. We found that in the spleen and liver MIP-1alpha, MIP-2 and Mig were the predominant chemokines expressed. In another target organ, the skin, MIP-1alpha, MIP-2, MCP-1 and MCP-3 were all highly expressed. In a non-target organ of acute GVHD, the heart, the predominant chemokines expressed were MCP-1 and MCP-3. This distinct pattern of chemokine expression in these organs may contribute to the preferential recruitment of inflammatory cells into the liver and skin, but not into the heart, in acute GVHD.

Immunoreactivity of Hu proteins facilitates identification of myenteric neurones in guinea-pig small intestine.

Hu proteins, together with neurone-specific enolase (NSE), protein gene product 9.5 (PGP-9.5), microtubule-associated protein-2 (MAP-2) and tubulin beta III isoform, were evaluated immunohistochemically as neuronal markers in whole-mount preparations and cultures obtained from the myenteric plexus of guinea-pig small intestine. Anti-Hu immunostaining marked the ganglion cell somas and nuclei without staining of the neuronal processes in the whole-mounts and cultures. The ganglion cell bodies were not obscured by staining of multiple neuronal fibres and this facilitated accurate counting of the neurones. MAP2 immunostaining also provided clear images of individual neurones in both whole mounts and cultures. Immunoreactivity for NSE, PGP-9.5 and tubulin beta III isoform provided sharp images of the ganglion cells in culture, but not in whole-mount preparations. Strong staining of the neuronal processes in the whole-mount preparations obscured the profiles of the ganglion cell bodies to such an extent that accurate counting of the total neuronal population was compromised. Anti-Hu immunostaining was judged to be an acceptable method for obtaining reliable estimates of total numbers of myenteric neurones in relation to other specific histochemical properties such as histamine binding.

P2X(7) receptors in the enteric nervous system of guinea-pig small intestine.

The P2X(7) purinergic receptor subtype has been cloned and emphasized as a prototypic P2Z receptor involved in neurotransmission in the central nervous system and ATP-mediated lysis of macrophages in the immune system. Less is known about the neurobiology of P2X(7) receptors in the enteric nervous system (ENS). We studied the distribution of the receptor with indirect immunofluorescence and used selective agonists and antagonists to analyze pharmacologic aspects of its electrophysiologic behavior as determined with intracellular "sharp" microelectrodes and patch-clamp recording methods in neurons identified morphologically by biocytin injection in the ENS. Application of ATP or 2'- (or-3'-) O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzBzATP) activated an inward current in myenteric neurons. Brilliant blue G, a selective P2X(7) antagonist, suppressed the responses to both agonists. Potency of the antagonist was greatest (smaller IC(50)) for the current evoked by BzBzATP. The P2X(7) antagonists 1-[N,O-bis (1,5-isoquinolinesulfonyl)-N-methyl-l-tyrosyl]-4-piperazine (KN-62) and oxidized ATP also suppressed the BzBzATP-activated current. Micropressure application of BzBzATP evoked rapidly activating depolarizing responses in intracellular studies with "sharp" microelectrodes. Oxidized-ATP suppressed these responses in both myenteric and submucosal neurons. Rapidly activating depolarizing responses evoked by application of nicotinic, serotonergic 5-HT(3), or gamma-aminobutyric acid A (GABA(A)) receptor agonists were unaffected by brilliant blue G. Immunoreactivity for the P2X(7) receptor was widely distributed surrounding ganglion cell bodies and associated with nerve fibers in both myenteric and submucous plexuses. P2X(7) immunoreactivity was colocalized with synapsin and synaptophysin and surrounded ganglion cells that contained either calbindin, calretinin, neuropeptide Y, substance P, or nitric oxide synthase. The mucosa, submucosal blood vessels, and the circular muscle coat also showed P2X(7) receptor immunoreactivity.

Galanin suppresses calcium conductance and activates inwardly rectifying potassium channels in myenteric neurones from guinea-pig small intestine.

Whole-cell patch-clamp recording methods were used to investigate the ionic mechanisms underlying the hyperpolarizing action of galanin in enteric neurones. Galanin suppressed calcium current (ICa) and activated inwardly rectifying potassium current (IK,ir) in AH-type myenteric neurones of guinea-pig small intestine. Both suppression of ICa and activation of IK,ir were concentration-dependent, with an EC50 of 1.4 nmol L-1 and 55 nmol L-1, respectively. Pretreatment with pertussis toxin eliminated both actions of galanin, suggesting that both galanin-induced inhibition of ICa and galanin-induced activation of IK,ir involved activation of Gi/Go proteins. Both suppression of ICa and activation of IK,ir by galanin were mimicked by the N-terminal fragment of galanin, galanin-(1-16) suggesting that the first 16 amino acids of the peptide were sufficient for both actions. The galanin receptor antagonist galantide suppressed the galanin-induced activation of IK,ir with an EC50 of 16 nmol L-1. However, galantide alone suppressed ICa. The results suggest two mechanisms of action for galanin: one is opening of inwardly rectifying potassium channels and the second is blockade of voltage-activated calcium channels.

Pre- and postsynaptic inhibition by nociceptin in guinea pig small intestinal myenteric plexus in vitro.

Actions of nociceptin on electrical and synaptic behavior of morphologically and neurochemically identified neurons in the guinea pig duodenal myenteric plexus were studied with conventional techniques. Nociceptin hyperpolarized the membrane potential in 104 of 121 AH-type and 28 of 51 S-type neurons with an EC(50) of 11.9 +/- 1.2 nM. Increased K(+) conductance accounted for the hyperpolarizing responses that were blocked by pertussis toxin and unaffected by naloxone. The selective opioid receptor-like (ORL)(1) receptor antagonist [Phe(1)-psi(CH(2)-NH)-Gly(2)]nociceptin(1--13)-NH(2) suppressed the nociceptin-evoked responses while behaving like a partial agonist. The nonselective ORL(1) antagonist naloxone benzoylhydrazone competitively suppressed nociceptin actions with a pA(2) value of 5.8. Nociceptin acted at presynaptic inhibitory receptors to suppress fast excitatory nicotinic postsynaptic potentials in 25 of 30 neurons (EC(50) = 22.5 +/- 4.4 nM) and slow synaptic excitation in 38 of 45 neurons (EC(50) = 15.1 +/- 1.6 nM). Presynaptic inhibitory action of nociceptin was unaffected by naloxone and was antagonized by [Phe(1)-psi(CH(2)-NH)-Gly(2)]nociceptin(1--13)-NH(2) or naloxone benzoylhydrazone. The results suggest that nociceptin acts both pre- and postsynaptically by activating an ORL(1) receptor that is distinct from typical naloxone-sensitive opioid receptors.