[Advances in immunotherapy for pulmonary sarcomatoid carcinoma].

Pulmonary sarcomatoid carcinoma (PSC) is a rare, poorly differentiated non-small cell lung cancer (NSCLC) that contains sarcomatoid components or sarcomatoid differentiation, and accounts for less than 1% of all lung tumors. Compared to other types of NSCLC, PSC has more invasive biological behavior, is prone to metastasis, and has a higher recurrence rate after early surgery. Its greater resistance to traditional treatments leads to a poorer prognosis compared to other NSCLCs. Immunotherapy offers the possibility of long-term survival for PSC patients.

[Analysis of risk factors influencing the detection rate of urate crystal by dual energy computed tomography].

OBJECTIVE: To explore the risk factors of detection of uric acid crystals by dual energy CT (DECT) in patients with gout diagnosed by gold standard. METHODS: From June 2011 to December 2018, clinical data of 29 patients were collected who were diagnosed with acute or chronic gout by positive polarized light analysis of joint synovial fluid in First Hospital of Peking University. Chi-square test, Logistic regression and t-test were used. The relationship between DECT and the clinical data, laboratory examination and drug treatment were analyzed. RESULTS: In this study, 29 patients were included, of whom, 22 patients were detected with uric acid crystals by DECT, and 7 patients were not. According to whether the uric acid crystals were detected or not by DECT, the patients were divided into two groups. Compared with the negative group, the patients were older in positive group [(47±12) vs. (39±11) years, P=0.15], had higher body bass index (BMI) [(27.9±3.7) vs. (22.8±2.1) kg/m2, P=0.002], longer gout disease duration [(135±102) vs.(45±53) months, P=0.035], higher in the highest serum uric acid in history [(643±121) vs. (543±103) µmol/L, P=0.043]. Although uric acid near DECT in positive group was higher than in negative group, there was no statistical difference [(558±150) vs. (513±89) µmol/L, P=0.497]. Comparing positive group with negative group, the percentage of the patients in acute phase was higher than in chronic phase [18(81.8%) vs. 4(57%), P=0.311];the percentage of the patients taking uric-acid-lowering drugs was higher than the other group [22(100%) vs. 5 (71%), P=0.052];the percentage of the patients with recurrent typical attacks was higher than that of those without typical attacks [22 (100%) vs.6 (85%), P=0.241]. The consistency of symptoms and the finding of uric acid crystals by DECT had been compared between the joints. The right knee joint had the highest consistency (Kappa=0.627), followed by the left MTP1 (Kappa=0.58), the right metatarsophalangeal 1(MTP1, Kappa=0.551) and the left knee (Kappa=0.494), all of which had statistical significance. The consistency of the ankle joint was lower (the right ankle joint: Kappa=0.19, the left ankle joint: Kappa=0.256), showing no statistical significance. BMI (kg/m2) [2.307 (1.139-4.670), P=0.02], gout duration (years) [0.306 (0.906-4.881), P=0.186], and the highest uric acid level in history (mg/dL) [0.023 (0.981-2.764), P=0.137] had relationship to the positive result of urate crystals in DECT. CONCLUSION: Gout patients with larger BMI, higher previous highest uric acid value and longer gout duration had higher sensitivity of the positive result in DECT.

Tudor knockdown disrupts ovary development in Bactrocera dorsalis.

One of the main functions of the piwi-interacting RNA pathway is the post-transcriptional silencing of transposable elements in the germline of many species. In insects, proteins belonging to the Tudor superfamily proteins belonging to the Tudor superfamily play an important role in to play an important role in this mechanism. In this study, we identified the tudor gene in the oriental fruit fly, Bactrocera dorsalis, investigated the spatiotemporal expressional profile of the gene, and performed a functional analysis using RNA interference. We identified one transcript for a tudor homologue in the B. dorsalis transcriptome, which encodes a protein containing the typical 10 Tudor domains and an Adenosine triphosphate (ATP) synthase delta subunit signature. Phylogenetic analysis confirmed the identity of this transcript as a tudor homologue in this species. The expression profile indicated a much higher expression in the adult and pupal stages compared to the larval stages (up to a 60-fold increase), and that the gene was mostly expressed in the ovaries, Malpighian tubules and fat body. Finally, gene knockdown of tudor in B. dorsalis led to clearly underdeveloped ovaries in the female adult and reductions in copulation rate and amount of oviposition, indicating its important role in reproduction. The results of this study shed more light on the role of tudor in ovary development and reproduction.

Association of electromyographic activation patterns with pain and functional disability in people with chronic neck pain.

PURPOSE: This study examined the activation patterns of the cervical and thoracic muscles in people with and without chronic neck pain during functional activities and their associations with pain intensity and functional disability. METHODS: Thirty-four adults with chronic neck pain and 34 asymptomatic adults were recruited. They were requested to perform active cervical movements and an upper limb lifting task. Electromyographic activation patterns (EMG) of bilateral upper trapezius, cervical erector spinae, sternocleidomastoid, and thoracic erector spinae were recorded during these tasks. Correlation and multiple regression analysis were used to examine the associations between EMG variables and severity of pain and functional disability. RESULTS: When performing the cervical movements, the neck pain group displayed lower EMG activity levels, especially in the cervical and thoracic extensors. In addition, significantly prolonged activation was observed in seven of the ten muscles during the upper limb lifting task. The changes in EMG amplitude and activation duration were found to be significantly correlated with severity of pain (R2 = 0.716) and functional disability (R2 = 0.623). CONCLUSIONS: Significant differences in the activation patterns of multiple cervical and thoracic muscles were found in individuals with neck pain compared with those without neck pain. These were significantly associated with their degree of pain and functional limitation. The findings of this study highlight the importance of assessing and optimising the neuromuscular activation of these muscles in the rehabilitation of those suffering from chronic neck pain.

[Outcomes after singleversus bilateral lung transplantation for idiopathic pulmonary fibrosis].

OBJECTIVE: To compare outcomes after single and bilateral lung transplantation in patients with idiopathic pulmonary fibrosis (IPF). METHODS: 104 patients with IPF who underwent lung transplantation at Wuxi People's Hospital from January 2008 to February 2015 were evaluated. Recipients were divided into single lung transplantation (STL)group(41cases)and bilateral lung transplantation(BLT) group (63 cases). Preoperative characteristics, postoperative outcomes, postoperative complications, functional outcomes, survival and causes of death between the two groups were analyzed retrospectively. The patients were followed up by letter, call and outpatient visit. RESULTS: Preoperative characteristics: The SLT group were significantly older (P<0.05), and had significantly higher systolic (P<0.05) pulmonary artery pressures than the BLT group. Postoperative outcomes: The intraoperative blood loss of SLT group as significantly less than BLT group (P<0.05). The cold ischemic time of SLT group as significantly shorter than BLT group (P<0.05). The BLT group was significantly more frequently required extracorporeal membrane oxygenation (ECMO) support than SLT group (P<0.05). There was no statistically significant difference inventilator support, reoperation, reintubation, length of ICU stay and death between the SLT and the BLT group (P>0.05). Postoperative complications: The BLT group had a higher incidence of primary graft dysfunction than the SLT group (P<0.05). There was no statistically significant difference in chest complications, acute rejection, airway complications, pulmonary infection between the SLT and the BLT group (P>0.05). Functional outcomes: FVC%, FEV1%, DLCO and 6-MWD, were significantly higher in the BLT group than the SLT group (P<0.05). SURVIVAL: The overall survival rates at 1, 2 and 3 years were not different between the SLT and the BLT group (75.3%, 61.6% and 46.2% vs 79.1%, 68.6% and 53.4%, P>0.05). CONCLUSIONS: The lung transplantationis an effective treatment for idiopathic pulmonary fibrosis, BLT could significantly improve the postoperative lung functionin comparison with STL, BLT may be a preferable operation for younger patients, especially if significant pulmonary hypertension is present.The short-term survival of BLT was similar to SLT, but long-term survivalstill needs further follow-up. Considering the quality of life, BLT is better than SLT.

[Analyzing the relevant factors affecting the clinical outcome of periodontal regeneration surgery].

Periodontitis can lead to periodontal tissue defect, tooth mobility and loss, which seriously affects the quality of life. Periodontal regeneration surgery is an important treatment method for repairing periodontal defects, and it is also the hotspot of current periodontal clinical and basic research. A comprehensive understanding of the factors affecting the efficacy of periodontal regenerative surgery can improve clinicians' periodontal treatment concepts, increases the predictability of treatment results, and enhances the level of clinical diagnosis and periodontal treatment. In order to instruct the clinicians, this article will explain the basic principles of periodontal regeneration and the key points of periodontal wound healing, and analyze the elements of periodontal regeneration surgery, which including the patient-related factors, local factors, surgical factors and regenerative material selection.

[Association between periodontitis and mild cognitive impairment: a clinical pilot study].

Objective: To evaluate the association between periodontitis and mild cognitive impairment (MCI), and explore the potential local oral risk factors for MCI. Methods: The study included 70 middle-aged and elderly subjects (44 females and 26 males) with periodontal disease who were first diagnosed by the Department of Periodontology or referred by the Department of Geriatrics in Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to January 2022. In this study, the control group consisted of periodontal disease patients without cognitive impairment, and the case group (MCI group) consisted of those diagnosed with MCI referred by the geriatrics specialists. Full-mouth periodontal examinations of all subjects were performed and periodontal indicators were recorded by periodontists, while digital panoramic radiographs were taken. The severity of periodontitis was defined according to the 1999 classification, and the staging and grading of periodontitis were defined according to the 2018 American Academy of Periodontology and European Federation of Periodontology classification. The mini-mental state examination scale was chosen by geriatricians to evaluate the cognitive function of the included subjects. The cubital venous blood was drawn to detect the expression levels of inflammatory factors such as hypersensitive C-reactive protein (hs-CRP), interleukin (IL)-1ß, IL-6 and tumor necrosis factor-α(TNF-α) in serum. Independent-samples t test and chi-square test were used to analyze the differences in population factors, periodontal-related indexes and serum inflammatory factors between the two groups (α=0.05). Odds ratios (OR) for MCI according to the severity of periodontitis and main periodontal clinical indexes were calculated by binary Logistic analysis. Results: Thirty-nine subjects were included in the control group and thirty-one in the MCI group. The age of the study population was (58.3±6.2) years (range: 45-70 years). The comparison between two groups showed that the control group was with higher educational background (χ²=9.45, P=0.024) and 2.6 years younger than the MCI group [(57.1±6.0) years vs. (59.7±6.3) years, t=-1.24, P=0.082]. The number and proportion of moderate to severe periodontitis in control group were significantly lower compared to those in MCI group (17 cases with 43.6% vs. 23 cases with 74.2%, χ²=6.61, P=0.010), and the OR of moderate to severe periodontitis adjusted by age and educational background was 3.00 (95%CI: 1.01-8.86, P=0.048). Compared with the grading (χ²=5.56, P=0.062) of periodontitis, staging had a greater impact on MCI (χ²=7.69, P=0.041), moreover the proportion of MCI in stage Ⅰ grade A periodontitis was significantly lower than any other type of periodontitis (χ²=13.86, P=0.036). In addition, less presence of deep periodontal pockets [probing depth (PD)≥6 mm] (17.9% vs. 41.9%, χ²=4.87, P=0.027), fewer number of PD≥4 mm (6.48±6.70 vs. 11.03±8.91, t=-2.44, P=0.017), lower plaque index (1.42±0.56 vs. 1.68±0.57, t=-1.91, P=0.059) and gingival index (1.68±0.29 vs. 1.96±0.30, t=-3.93, P<0.001) were in the control group than in the MCI group. However, there were no significant differences between the two groups in the levels of serum inflammatory factors, such as hs-CRP, IL-1ß, IL-6 and TNF-α (P>0.05). Conclusions: It appears a strong correlation between moderate to severe periodontitis and the incidence of MCI in middle-aged and elderly people. Moreover, deep and increased number of periodontal pockets, poor oral hygiene, and severe gingival inflammation can be potentially associated risk factors for MCI.

Eomesodermin, HAND1, and CSH1 proteins are induced by cellular stress in a stress-activated protein kinase-dependent manner.

Eomesodermin (Eomes) is a transcription factor essential for trophoblast development. Stress stimuli activate stress-activated protein kinase (MAPK8/9) and modulate transcription factors in trophoblast stem cells (TSC). In this study, we test the hypothesis that stress-induced Eomes upregulation and downstream trophoblast development are MAPK8/9-dependent. Immunocytochemical and immunoblot assays suggest that Eomes is induced by hyperosmolar stress in a dose- and time-dependent manner. Two MAPK8/9 inhibitors that work by different mechanisms, LJNKl1 and SP600125, block induction of Eomes protein by stress. During normal TSC differentiation, the transcription factor heart and neural crest derivatives expressed 1 (HAND1) is dependent on Eomes, and chorionic somatomammotropin hormone 1 (CSH1) expression is dependent on HAND1. Similar to Eomes, HAND1 and CSH1 induction by stress are MAPK8/9-dependent, and CSH1 is induced in nearly all stressed TSC. CSH1 induction normally requires downregulation of the transcription factor inhibitor of differentiation 2 (ID2) as well as HAND1 upregulation. It was shown previously that hyperosmolar stress induces AMP-activated protein kinase (PRKAA1/2)-dependent ID2 loss in a MAPK8/9-independent manner. Inhibition of PRKAA1/2 with compound C and LJNKl1, more than MAPK8/9 inhibitors alone, inhibits the induction of CSH1 by stress. Taken together these data suggest that stress-induced MAPK8/9 and PRKAA1/2 regulate transcription factors Eomes/HAND1 and ID2, respectively. Together this network mediates induction of CSH1 by stress. Therefore, stress triggers a proportional increase in a normal early TSC differentiation event that could be adaptive in inducing CSH1. But the flexibility of TSC to undergo stress-induced differentiation could lead to pathophysiological consequences if stress endured and TSC differentiation became unbalanced.

[Periodontally accelerated osteogenic orthodontics in clinical treatment].

Orthodontic tooth movement inevitably affects adjacent periodontal tissue. The labiobuccal alveolar bone is generally thin, and patients with dental crowding often present alveolar dehiscence and fenestration. These may lead to prolonged treatment time, even gingival atrophy, tooth loosening and other complications in the orthodontic tooth movement. Periodontally accelerated osteogenic orthodontics (PAOO) could accelerate the orthodontic tooth movement and promote the formation of new bone by using corticotomy and bone grafting. This article, starting from the influence of orthodontic treatment on periodontal hard tissue, describes the application of PAOO in orthodontic treatment.

Curcumin improves experimentally induced colitis in mice by regulating follicular helper T cells and follicular regulatory T cells by inhibiting interleukin-21.

To determine whether curcumin (Cur) can treat mice with experimentally-induced colitis by regulating follicular helper T cells (Tfh) and follicular regulatory T cells (Tfr) by inhibiting interleukin (IL)-21. In this study, 40 male C57BL/6 mice were randomly grouped into four groups, i.e., normal, trinitrobenzene sulfonic acid (TNBS), TNBS + curcumin, and TNBS + anti-IL-21. Mice with experimental colitis were induced by 100 mg/kg TNBS. The mice in the TNBS + Cur group were treated with 100 mg/kg curcumin for seven days, and mice in the TNBS + anti-IL-21 group were treated with anti-IL-21 (150 µg/mouse) once per week, intraperitoneally, starting on the second day after establishing the experimental colitis model. On day eight, the therapeutic effect of curcumin was evaluated by colon mucosa damage index (CMDI), histological examination, and disease activity index (DAI). Furthermore, the number of CD4 + CXCR5 + PD-1 + Tfh and CD4 + CXCR5 + FoxP3 + Tfr cells were measured by flow cytometry. The mRNA and protein expression of IL-21, Bcl-6, FOXP3, ICOS, and PD-1 in colonic mucosa was detected by reverse transcription polymerase chain reaction and the Western blot technique. Compared with the TNBS group, the DAI, CMDI, histological score, the number of CD4 + CXCR5 + PD-1 + Tfh cells, the expression of IL-21, Bcl-6, ICOS, and PD-1 were significantly decreased in the TNBS + curcumin group and TNBS + anti-IL-21 group; body weight, number of CD4 + CXCR5 + FoxP3 + Tfr cells, and the expression of FoxP3 were observably elevated in the TNBS + curcumin group (all P < 0.05). Curcumin may have a potential therapeutic effect on mice with colitis treated experimentally through regulation of the balance of Tfh and Tfr cells via inhibiting the synthesis of IL-21.

Initial comparison of proteomic profiles of whole unstimulated saliva obtained from generalized aggressive periodontitis patients and healthy control subjects.

BACKGROUND AND OBJECTIVE: Salivary proteomics technology can be used to evaluate the disease progression of periodontitis and the systemic screening of proteomes of saliva from subjects with aggressive periodontitis has not been available. The objective of this preliminary study was to compare the proteomic profile of whole unstimulated saliva of subjects with generalized aggressive periodontitis (GAgP) with that of healthy volunteers to identify proteins, the levels of which were significantly altered between the two groups. MATERIAL AND METHODS: Whole unstimulated saliva was obtained from five subjects with GAgP and five healthy subjects, and proteins were separated using two-dimensional gel electrophoresis. Proteins, the levels of which were significantly different between the two groups, were identified by computer image analyses and subsequent electrospray ionization tandem mass spectrometry. RESULTS: Eleven proteins that exhibited a different level in the GAgP group vs. the control group were identified. Compared with whole saliva of healthy control subjects, the levels of serum albumin, immunoglobulin (Ig) gamma2 chain C region, Ig alpha2 chain C region, vitamin D-binding protein, salivary alpha-amylase and zinc-alpha2 glycoprotein were increased in whole unstimulated saliva of GAgP subjects, while those of lactotransferrin, elongation factor 2, 14-3-3 sigma, short palate, lung and nasal epithelium carcinoma-associated protein 2 precursor and carbonic anhydrase 6 were decreased. CONCLUSION: Comparison of the proteomic profile of whole unstimulated saliva of GAgP subjects with that of healthy control subjects revealed at least 11 differential proteins. The approach applied herein might be helpful to aid understanding of the etiology of GAgP.

Changes in the range of the medicinal herb Eriocaulon buergerianum Körnicke. (Eriocaulaceae) under climate change.

Eriocaulon buergerianum Körnicke. (Eriocaulaceae) is one of the most common and least expensive herbal medicines for eye disease. This species is facing potential threats from climate change. Insufficient biogeographic knowledge of this plant species can hinder its effective management for long-term population survival. We integrated ecological niche modelling (Biomod2) with 70 records of E. buergerianum and eight environmental variables to estimate changes in distribution over time. A core area Zonation algorithm was introduced to identify conservation priority areas. Our results indicate that the range of E. buergerianum will likely decrease in the future: the overall range change on average is -44.36 ± 21.56% (-3.70% to -77.73%); values of range loss and range gain are 45.79 ± 20.30% (9.29-78.19%) and 1.43 ± 1.53% (0.18-5.59%), respectively. According to conservation priority analysis, the mandatory reserve (top 5%), negotiable reserve (0.95-0.9) and partial reserve (0.9-0.8) areas are 19,799, 19,799 and 39,597 km2 , respectively. The areas identified as conservation priority are located in the southeast, especially in northern Taiwan and the Wuyi Mountains. Based on these results, we suggest a re-evaluation of the threatened status of this species, with a potential upgrade to the vulnerable (VU) category. To overcome the adverse conditions faced by populations of E. buergerianum in China, we propose a multi-faceted conservation strategy involving more complete resource assessment, a monitoring system, medical research focused on revealing medicinal components or substitutes, and a regional development plan that considers both wildlife and socio-economic issues.

[Expression profile of long non-coding RNA in gingival tissues from the patients with aggressive periodontitis].

Objective: To detect and analyze the differential expression profile of long non-coding RNA (lncRNA) in aggressive periodontitis (AgP) and healthy gingival tissues, in order to explore the role of lncRNA in AgP. Methods: After the informed consents were obtained, gingival tissues from AgP patients (n=40) and healthy volunteers (n=40) were collected in Department of Periodontology, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine (from Mar. 2012 to Aug. 2012) and Department of Periodontology, Hospital of Stomatology, Tianjin Medical University (from Oct. 2016 to Apr. 2017). The differential expression of lncRNA of tissues from AgP patients (n=20) and healthy volunteers (n=20) were examined via microarray assay. Bioinformatics was applied to analyze the expression data of lncRNA and correlative mRNA. Two lncRNAs (lncRNA-TNFRSF13C and lncRNA-API5) were chosen to verify the microarray results by using real time quantitative polymerase chain reaction (PCR) in the other gingival tissues. Results: Compared with the result of healthy gingival tissues, totally 8 632 lncRNAs were differentially expressed in tissues from AgP patients. From these data, 1 986 lncRNAs were significantly upregulated while 6 646 lncRNAs were downregulated, amongst which 48 lncRNAs were upregulated (>10 times) (P<0.05), 14 lncRNAs were downregulated (>10 times) (P<0.05). Furthermore, totally 5 519 correlative mRNAs were differentially expressed, amongst which 1 676 mRNAs were upregulated (≥2 times, P<0.05) and 3 843 mRNAs were downregulated≤0.5 (P<0.05). The selected lncRNA-TNFRSF13C and lncRNA-API5 were up-regulated in AgP (P<0.05), which confirmed the results of microarray. From bioinformatics, differential expression lncRNAs were in association with many signal pathways including toll-like receptor signaling pathway, cell cycle and apoptosis pathway, and tumor necrosis factor receptor superfamily pathway. Conclusions: LncRNA may be involved in the pathogenesis of AgP through various pathways, which need to be further explored.

A study of enamel matrix proteins on differentiation of porcine bone marrow stromal cells into cementoblasts.

OBJECTIVE: To further explore the role of enamel matrix proteins (EMPs) in periodontal regeneration, we have used porcine bone marrow-derived stromal cells (BMSCs) to observe whether the EMPs could have an effect on their differentiation into cementoblasts. MATERIALS AND METHODS: In this study, EMPs were extracted from porcine tooth germs by the use of acetic acid. BMSCs obtained from porcine iliac marrow aspiration were inoculated onto the surface of autologous root slices treated with or without EMPs. Following 7-day co-culture, all the BMSC-seeded root slices, with their respective non-cell-inoculated control specimens, were pocketed with expanded polytetrafluoroethylene membrane and were transplanted subcutaneously into 11 nude mice. The animals were sacrificed after 3 and 8 weeks, and the new specimens were processed for haematoxylin and eosin staining. RESULTS: Histological analysis demonstrated new cellular cementum-like tissue formed along EMP-treated root slices. CONCLUSION: Our work has indicated for the first time, differentiation of BMSCs into cementoblasts using an EMP-based protocol.

Central sensitization in thalamic nociceptive neurons induced by mustard oil application to rat molar tooth pulp.

We have recently demonstrated that application of mustard oil (MO), a small-fiber excitant and inflammatory irritant, to the rat maxillary molar tooth pulp induces central sensitization that is reflected in changes in spontaneous activity, mechanoreceptive field (RF) size, mechanical activation threshold, and responses to graded mechanical stimuli applied to the neuronal RF in trigeminal brainstem subnucleus caudalis and subnucleus oralis. The aim of this study was to test whether central sensitization can be induced in nociceptive neurons of the posterior thalamus by MO application to the pulp. Single unit neuronal activity was recorded in the ventroposterior medial nucleus (VPM) or posterior nuclear group (PO) of the thalamus in anesthetized rats, and nociceptive neurons were classified as wide dynamic range (WDR) or nociceptive-specific (NS). MO application to the pulp was studied in 47 thalamic nociceptive neurons and found to excite over 50% of the 35 VPM neurons tested and to produce significant long-lasting (over 40 min) increases in spontaneous activity, cutaneous pinch RF size and responses to graded mechanical stimuli, and a decrease in threshold in the 29 NS neurons tested; a smaller but statistically significant increase in mean spontaneous firing rate and decrease in activation threshold occurred following MO in the six WDR neurons tested. Vehicle application to the pulp did not produce any significant changes in six VPM NS neurons tested. MO application to the pulp produced pronounced increases in spontaneous activity, pinch RF size, and responses to mechanical stimuli, and a decrease in threshold in three of the six PO neurons. In conclusion, application of the inflammatory irritant MO to the tooth pulp results in central sensitization of thalamic nociceptive neurons and this neuronal hyperexcitability likely contributes to the behavioral consequences of peripheral inflammation manifesting as pain referral, hyperalgesia and allodynia.

Mu but not delta and kappa opioid receptor involvement in ventrolateral orbital cortex opioid-evoked antinociception in formalin test rats.

The present study was designed to investigate the roles of different subtypes of opioid receptors in ventrolateral orbital cortex (VLO) opioid-evoked antinociception in formalin test by using an automatic detection system for recording the nociceptive behavior (agitation) and a manual method for detecting the duration of licking the injected paw in the conscious rat. Formalin (5%, 50 microl) s.c. injected into the hindpaw produced a biphasic agitation response or lengthening duration of licking. Morphine (5 microg) microinjected unilaterally into VLO significantly inhibited the agitation response and the licking time, and these effects were blocked by pre-administration of the non-selective opioid receptor antagonist naloxone (1.0 microg) into the same site. Microinjection of endomorphin-1 (5 microg), a selective micro-receptor agonist, and [D-Ala2, D-Leu5]-enkephalin (DADLE, 10 microg), a delta-/micro-receptor agonist also inhibited the nociceptive behaviors, and both the effects were blocked by selective mu-receptor antagonist beta-funaltrexamine hydrochloride (beta-FNA; 3.75 microg), but the DADLE-evoked inhibition was not influenced by the selective delta-receptor antagonist naltrindole (5 microg). Microinjection of selective kappa-receptor agonist (+/-)-trans-U-50488 methanesulfonate salt (1.5 microg) failed to alter the nociceptive behaviors induced by formalin injection. The beta-FNA and naloxone applied into VLO and morphine into the adjacent regions ventral and dorsal to VLO had no effect on the formalin-evoked nociceptive behaviors. These results suggest that mu- but not delta- or kappa-opioid receptor is involved in the VLO opioid-evoked antinociception in formalin test rat.

Serotonergic, noradrenergic and galaninergic projections to the nucleus parafascicularis.

Immunocytochemical staining for serotonin (5-HT), tyrosine hydroxylase (TH) and galanin (GAL) was combined with horseradish peroxidase (HRP) retrograde tract-tracing technique to analyze the localizations of 5-HT-, catecholamine (CA)- and GAL-containing neurons in the brainstem which project to the nucleus parafascicularis (PF) in rats. It is demonstrated that most of the retrogradely HRP-labeled neurons (70%) in bilateral periaqueductal gray (PAG) and raphe nuclei are positively immunostained by antiserum to 5-HT, and that most of the retrogradely HRP-labeled neurons (over 80%) in bilateral locus coeruleus (LC) are positively immunostained by antisera to both TH and GAL. The possible functions of these PF-petal serotonergic, catecholaminergic (actually noradrenergic) and galaninergic projections are discussed.

Morphine and norepinephrine-induced antinociception at the spinal level is mediated by adenosine.

The purpose of this study was to examine whether adenosine or serotonin is involved in mediation of the antinociception produced by norepinephrine at the spinal cord level. Aminophylline, an adenosine receptor antagonist and naloxone given intrathecally (i.t.) were used to test the antinociception produced by i.t. norepinephrine, serotonin, morphine or the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) by using the tail-flick assay in rats. It was observed that (1) aminophylline blocked the antinociception produced by norepinephrine, but exhibited no effect on the antinociception produced by serotonin, (2) aminophylline blocked the antinociception produced by morphine similarly to naloxone, (3) aminophylline blocked the antinociception produced by NECA and (4) naloxone failed to block the antinociception produced by NECA and serotonin. The results suggest that adenosine is involved in mediation of the norepinephrine-produced antinociception at the spinal level and that norepinephrine and adenosine may act in a sequential manner in norepinephrine-induced antinociception.