IL-38: A novel cytokine in systemic lupus erythematosus pathogenesis.

IL-38 is a newly identified cytokine that belongs to the IL-1 family. In our previous study, we found elevated plasma levels of IL-38 in patients with systemic lupus erythematosus (SLE). However, the clear relationship of IL-38 expression in plasma, peripheral blood mononuclear cells (PBMCs) and clinical and laboratory features needs elucidation. Additionally, we evaluated the possible role of IL-38 in regulating production of inflammatory cytokines in PBMCs in vitro. A pristane-induced murine lupus model was used to further demonstrate the effects of IL-38 on cytokines in vivo and discuss the significance of IL-38 in lupus development. The results showed that mRNA expression of IL-38 in PBMCs of patients with SLE was elevated compared with volunteers, and expression of IL-38 in both plasma and PBMCs was strongly related to clinical features, such as haematuria and proteinuria, and correlated with a SLEDAI score. Plasma levels of TNF-α, IL-1ß, IL-6 and IL-23 were elevated in patients with SLE and were related to plasma levels of IL-38. In vitro, PBMCs of patients with SLE stimulated with IL-38 showed a decreased expression of the four inflammatory cytokines compared with PBMCs of patients without treatment. Interestingly, IL-38 administration in lupus mice significantly reduced the development of lupus, such as reduced proteinuria, improved histological examinations of the kidneys and down-regulated inflammatory cytokines. In conclusion, IL-38 may suppress synthesis of pro-inflammatory cytokines and therefore regulate lupus pathogenesis.

Homozygous ALOXE3 Nonsense Variant Identified in a Patient with Non-Bullous Congenital Ichthyosiform Erythroderma Complicated by Superimposed Bullous Majocchi's Granuloma: The Consequences of Skin Barrier Dysfunction.

Non-bullous congenital ichthyosiform erythroderma (NBCIE) is a hereditary disorder of keratinization caused by pathogenic variants in genes encoding enzymes important to lipid processing and terminal keratinocyte differentiation. Impaired function of these enzymes can cause pathologic epidermal scaling, significantly reduced skin barrier function. In this study, we have performed a focused, genetic analysis of a probrand affected by NBCIE and extended this to his consanguineous parents. Targeted capture and next-generation sequencing was performed on NBCIE associated genes in the proband and his unaffected consanguineous parents. We identified a homozygous nonsense variant c.814C>T (p.Arg272*) in ALOXE3 (NM_001165960.1) in the proband and discovered that his parents are both heterozygous carriers of the variant. The clinical manifestations of the proband's skin were consistent with NBCIE, and detailed histopathological assessment revealed epidermal bulla formation and Majocchi's granuloma. Infection with Trichophyton rubrum was confirmed by culture. The patient responded to oral terbinafine antifungal treatment. Decreased skin barrier function, such as that caused by hereditary disorders of keratinization, can increase the risk of severe cutaneous fungal infections and the formation of Majocchi's granuloma and associated alopecia. Patients with NBCIE should be alerted to the possible predisposition for developing dermatophytoses and warrant close clinical follow-up.

[IgG subclasses targeting NC16A domain of BP180 in bullous pemphigoid].

OBJECTIVE: To evaluate the distribution of four IgG subclasses targeting NC16A domain of BP180 in bullous pemphigoid (BP) patients by developing and optimizing a detection method of anti-BP180NC16A IgG subclasses so as to assess its sensitivity and specificity. METHODS: Enzyme-linked immunosorbent assay (ELISA) was developed with recombinant GST-BP180NC16A proteins generated by a bacterial expression system. And 136 BP sera and 20 healthy control sera from our hospital between 2009 and 2012 were tested by ELISA, and the cutoff value of four IgG subclasses was set at an A reading corresponding to the mean value plus 3 times of standard deviation of 20 healthy controls sera. Western blot was also used to detect the IgG subclasses in patients with four positive IgG subclasses by ELISA. RESULTS: The cutoff value of specific IgG1, IgG2, IgG3 and IgG4 were 0.113,0.196,0.154 and 0.120. The values of four IgG subclasses from 20 healthy controls were lower than the corresponding cutoff value, making the detection system good specificity. The positive rates of anti-BP180NC16A IgG1, IgG2, IgG3 and IgG4 antibody were 67.6% (92/136) , 45.6% (62/136), 50.7% (69/136) and 54.4% (74/136) respectively in 136 BP sera. All four IgG subclasses were positive in 29 BP sera, accounting for 21.3%. The number of BP sera positive for at least one IgG subclass were 112, accounting for 82.4%, indicating that the combined sensitivity of four IgG subclasses was 82.4%. Western blot revealed that the number of positivity was 15 and 14 for IgG1 and IgG4 respectively in 20 BP patients with four IgG subclasses positive with ELISA. CONCLUSION: The specificity of ELISA is excellent while its sensitivity needs further improvements.

[Establishment of enzyme-linked immunosorbent assay in the detection of BP180NC16A-specific IgE and its significance in bullous pemphigoid].

OBJECTIVE: To establish a method of detecting circulating immunoglobulin E (IgE) autoantibodies for BP180NC16A and evaluate its significance in bullous pemphigoid (BP). METHODS: GST-NC16A fusion proteins were expressed in Escherichia coli using the pGEX-2T expression system and purified by glutathione affinity chromatography.For optimal working conditions of enzyme-linked immunoabsorbent assay (ELISA), checkerboard titrations were performed with serial dilutions of antigen. Also optimized dilution of secondary antibody was confirmed. Sera samples from 56 patients with BP, 24 healthy controls, 18 with pemphigus and 1 with Stevens-Johnson syndrome at our hospital during February 2011 to October 2012 were examined by the modified ELISA approach. The optimal cut-off point for a positive result was selected with receiver operating characteristic analysis. RESULTS: The optimized ELISA was performed with plates coated with 500 µg/ml GST-NC16A. And the optimal dilutions of sera samples and secondary antibody were 1: 10 and 1: 1000 respectively. According to the established cut-off value (0.549), 40 of 56 BP patients and none of controls had detectable levels of BP180NC16A IgE. CONCLUSION: The established ELISA provides a highly specific tool for the detection of IgE anti-BP180NC16A in BP patients.

Placental and maternal serum inhibin A in patients with preeclampsia and small-for-gestational-age.

AIM: The purpose of this study was to determine the levels of inhibin A simultaneously in the maternal serum and placental extract in preeclampsia (PE) with or without small-for-gestational-age (SGA) and normal controls at term, and to evaluate the relationship among changes in serum and placental inhibin A according to the severity of PE and PE with or without SGA. MATERIAL AND METHODS: This study involved 40 pregnant women; normal (n = 20), and PE (n = 20), the latter of who were classified into (i) mild (n = 10) and severe PE (n = 10); (ii) PE with SGA (n = 7) and without SGA (n = 13). Inhibin A concentrations were quantified by enzyme-linked immunosorbent assay (ELISA) in the maternal serum and placental extract. Inhibin-α subunit in the placenta was stained by immunohistochemistry (IHC), and its intensity was graded by a semiquantitative scoring method. RESULTS: There was a positive correlation in inhibin A concentrations between the serum and placental extract (r = 0.57, P < 0.001). Both maternal serum and placental inhibin A in PE groups were significantly higher than in controls, but there was no severity-dependent increase of inhibin A when compared with mild and severe PE. There was no difference in inhibin A levels between PE with and without SGA. Moreover, the inhibin-α subunit was predominantly abundant in the cytoplasm of the syncytiotrophoblasts, where the PE groups showed higher staining intensity than the controls (P < 0.000). CONCLUSION: Serum inhibin A level might be a useful biomarker for diagnosis and monitoring of PE.

Comparison of benthic nematode assemblages in native mangrove forest and exotic mangrove plantations (Sonneratia apetala Buch-Ham) along the South China Coast.

The exotic mangrove species Sonneratia apetala has been planted widely in China since 1985. As an exotic mangrove species, it is important to understand the influence of Sonneratia plantations on marine nematode assemblages. We studied the Zhanjiang and Xiatanwei mangrove wetlands on the South China Coast during the four seasons of 2010 and 2019. The results showed that S. apetala plantations increased the number of genera, Shannon-Wiener diversity, richness, evenness, maturity index (MI) and decreased the index of trophic diversity (ITD) values for nematodes compared to those in mudflats, but did not change significantly compared to those of native mangrove forests. In addition, Sonneratia plantations did not significantly change nematode assemblages compared to those of native mangroves, though they did change significantly compared to those in mudflats. Thus, interplanting S. apetala into native mangrove forests may be an effective substitute for using native mangroves in mangrove plantation.

Aberrantly increased mRNA expression of betaglycan, an inhibin co-receptor in the ovarian tissues in women with polycystic ovary syndrome.

AIM: To compare the gene expressional levels of receptors for activins and inhibins in ovarian tissues between women with polycystic ovary syndrome (PCOS) and normal controls, and to analyze their biologically relevant associations with serum hormone levels. METHODS: Total RNA of ovarian tissues from PCOS (n = 14) and normal controls (n = 21) were isolated during the follicular phase of the menstrual cycle. Real-time PCRs were performed to examine the relative mRNA expression levels of the activin receptors, including activin receptor type IA (ActRIA), type IB (ActRIB), type IIA (ActRIIA), and type IIB (ActRIIB), and the inhibin receptors, betaglycan and an inhibin binding protein (InhBP/p120). At the same time, the serum levels of estradiol (E(2)), testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B and activin A were measured. RESULTS: The PCOS patients showed endocrine characteristics with higher concentrations of LH, T and inhibin B, and a lower concentration of activin A. Real-time PCR demonstrated that the relative expression level of betaglycan against that of GAPDH was increased 1.5-fold in the ovarian tissues during the follicular phase of PCOS patients when compared with normal controls, while the activin receptors ActRIA, ActRIB, ActRIIA, ActRIIB, and the inhibin co-receptor InhBP/p120 were unchanged. Moreover, the betaglycan mRNA expression showed biologically relevant associations with serum FSH, LH, E(2) and inhibin B levels in both the PCOS and normal controls. CONCLUSIONS: This is the first report to demonstrate the aberrantly increased expression of betaglycan mRNA in PCOS ovaries. The mechanism by which betaglycan contributes to the pathologic process of PCOS remains to be clarified.

X-linked dominant protoporphyria in a Chinese pedigree reveals a four-based deletion of ALAS2.

BACKGROUND: X-linked dominant protoporphyria (XLDPP) is a rare, hereditary disorder that leads to hepatobiliary and hematologic abnormalities including increased erythrocyte protoporphyrin, cutaneous photosensitivity, and decreased iron stores that is caused by a pathogenic mutation of ALAS2 gene. METHODS: This study aimed to confirm the existence of XLDPP in a Chinese pedigree. We observed and described the dermatoscopic findings of this disorder under dermoscopy, and assessed photo damage in XLDPP patients using the Fotofinder system and very high frequency (VHF) skin ultrasonic system. We performed next generation sequencing and Sanger sequencing to detect and confirm genetic variants in DNA samples from the XLDPP family. Moreover, we monitored the hepatobiliary function as well as hematologic changes in related family members. RESULTS: As compared to unaffected control subjects, patients exhibited evidence of severe cutaneous photodamage, causing photoaging, an increase in the size of the gallbladder, increased levels of protoporphyrin in red blood cells, an increase in blood levels of uroporphyrin and hematoporphyrin, and iron deficiency. CONCLUSIONS: XLDPP was validated by the identification of a four-base-pair deletion (c.1706_1709delAGTG, p.E569fs) in ALAS2 (NM_000032.4) in the proband which segregated with the disease in an X-linked dominant pattern, with hemizygous males being more severely affected than heterozygous females. We also found a missense variant in GATA Binding Protein 1 (GATA1).

Inhibitory effect of crotoxin on the pain-evoked discharge of neurons in thalamic parafascicular nucleus in rats.

Crotoxin (Cro), the principal neurotoxic component of Crotalus durissus terrificus, has been previously reported to have a behavioral analgesic effect in rats and mice. The present study investigated electrophysiologically the effect of Cro on pain-evoked unit discharge of neurons in thalamic parafascicular nucleus (Pf) and underlying mechanisms of its effect. The electrical discharge of Pf neurons was recorded with the microelectrode technique in rats. Intracerebroventricular (i.c.v.) injection of Cro at 0.25, 0.45 and 0.65 microg/kg resulted in a dose-dependent inhibitory effect on the pain-evoked discharge of Pf neurons. The discharge frequency and the discharge duration significantly (P<0.05) decreased after Cro administration. This inhibitory effect was significantly (P<0.05) attenuated after pretreatment with para-chlorophenylalanine (pCPA), or electrolytic lesion of dorsal raphe (DR) nucleus. In contrast, i.c.v. injection of atropine (muscarinic receptor antagonist, 5 microg) or naloxone (opioid receptor antagonist, 4 microg) had no effect on Cro-induced inhibition of discharge of Pf neurons. The results suggested that Cro has an analgesic effect, which is mediated, at least partially, by the central serotonergic system.

Endoplasmic reticulum stress-mediated autophagy contributes to 5-ethylamino-9-diethylaminobenzo[a]phenoselenazinium-mediated photodynamic therapy via the PERK-eIF2α pathway.

INTRODUCTION: 5-ethylamino-9-diethylaminobenzo[a]phenoselenazinium (EtNBSe) is a novel synthetic bipolar photosensitizer with many promising applications. This study investigated the impact of EtNBSe-mediated photodynamic therapy (EtNBSe-PDT) on the autophagy and endoplasmic reticulum (ER) stress of squamous carcinoma cells (A-431 cells), as well as the related molecular mechanisms. METHODS: The potency of EtNBSe-PDT against squamous cell carcinoma was evaluated in BALB/c nude mice. Cell viability was evaluated using MTT. Western blotting and immunofluorescence were used to determine the expression levels of ER stress- and autophagy-related proteins. RESULTS: Both morphological and microscopic findings showed that the tumor on the xenograft mice exhibited an apparent reduction in volume and was replaced with fibrosis 20 days after EtNBSe-PDT. Additionally, in an in vitro study using A-431 cells, EtNBSe-PDT was found to inhibit A-431 cell survival in an EtNBSe concentration- and light dose- dependent manner, and to induce ER stress via the PERK-eIF2α signaling pathway. Additionally, EtNBSe-PDT could also induce autophagy of A-431 cells. Furthermore, the ER stress inhibitor 4-PBA and the eIF2α inhibitor salubrinal were found to inhibit the autophagy induced by EtNBSe-PDT. CONCLUSION: This study demonstrated that the PERK-eIF2α signaling pathway was involved in the ER stress induced by EtNBSe-PDT. Meanwhile, the ER stress via the PERK-eIF2α pathway promoted the occurrence of autophagy in A-431 cells.

Serum levels of immunoglobulins G1 and G4 targeting the non-collagenous 16A domain of BP180 reflect bullous pemphigoid activity and predict bad prognosis.

Bullous pemphigoid (BP) is an autoimmune subepidermal bullous disease, and different immunoglobulin (Ig)G autoantibody subclasses may play different roles in the pathogenesis of BP. This study aims to evaluate the relationship between specific IgG subclasses and BP. Enzyme-linked immunoassays (ELISA) were developed to test the IgG subclasses targeting the non-collagenous 16A (NC16A) domain of BP180. A statistical analysis was carried out to assess the relationship of BP and IgG subclasses as well as other factors. The correlation coefficients between the ELISA scores for four IgG subclasses and disease severity scores were 0.586 for IgG, 0.441 for IgG1, 0.594 for IgG2, 0.345 for IgG3, and 0.448 for IgG4 before treatment. After treatment, the correlation coefficient was 0.376 for IgG, 0.522 for IgG1, 0.314 for IgG2, 0.582 for IgG3 and 0.503 for IgG4. Spearman's rank correlation coefficient was 0.801 for IgG1, 0.66 for IgG2, 0.575 for IgG3 and 0.463 for IgG4 between the ELISA scores of IgG subclasses and the disease severity score variation. The ELISA scores of IgG subclasses in patients with mucosal involvement were higher than those without. Survival analysis showed that sex, IgG1 and IgG4 were the independent predictors for BP. In conclusion, the serum levels of IgG1 and IgG4 targeting BP180NC16A were paralleled with disease severity in BP patients. IgG1 and IgG4 and sex were the independent prognostic factors for an early prognosis of BP.

Incidence of adverse cutaneous drug reactions in 22,866 Chinese inpatients: a prospective study.

Cutaneous adverse drug reactions (ADRs) are common. However, no prospective study assessing cutaneous ADRs is available for Chinese populations. This study aimed to assess the incidence, manifestations, causative drugs, and other factors related to cutaneous ADRs. A total of 22,866 inpatients were surveyed prospectively from January to April 2012 at the Peking Union Medical College Hospital. Only cutaneous ADRs induced by systemic drugs were considered. Fifty cases were confirmed as cutaneous ADRs, for an estimated incidence of 2.2 per 1000 during this period (95 % confidence interval 1.6-2.8). Cases of cutaneous ADRs comprised 69 % females, while 63 % of all inpatients were female (χ (2) = 0.641, P = 0.427). The department of infectious diseases was the most frequently involved department. Morbilliform exanthema (40 %) was the most frequent cutaneous ADR, followed by urticaria (23.1 %). Anti-infection drugs (36.9 %) caused most cases of cutaneous ADRs, followed by iodinated contrast media (ICM, 18.5 %) and non-steroidal anti-inflammatory drugs (NSAIDs, 18.5 %). The most frequently associated disorders were cancer (24 %), infection (22 %), cardiovascular and cerebrovascular diseases (20 %), and autoimmune diseases (18 %). In this first prospective study assessing the incidence of cutaneous ADRs in China, anti-infection drugs were the most commonly involved drugs, followed by ICM and NSAIDs. No evidence of increased cutaneous ADR incidence in AIDS or SLE patients was observed. Our findings indicate that cancer and its treatments were often related to cutaneous ADRs in China.

[Effects of central histaminergic receptor activation on carotid sinus baroreceptor reflex resetting in stressed rats].

To determine the effect of stress on carotid sinus baroreceptor reflex (CSR) and whether or not central histaminergic receptors modulate the CSR under stress, the characteristics of CSR were analyzed by using an isolated carotid sinus preparation in Wistar rats. Animals were divided into two groups at random: unstressed group (n=42) and stressed group (n=41). According to the site of microinjection of histaminergic receptor antagonists, each group was subdivided into a group of intracerebroventricular injection (i.c.v.) and a group of microinjection into the nucleus of the solitary tract (NTS). The volume of injection into the lateral cerebroventricle and NTS was 5 microl and 1 micro1, respectively. Stressed groups were subjected to unavoidable electric foot-shock twice daily for a week, each session of foot-shock lasted 2 hours. The left and right carotid sinus regions were isolated from the systemic circulation under anesthesia with pentobarbital sodium in all rats. The intracarotid sinus pressure (ISP) was altered in a stepwise manner to trigger CSR from 0 to 280 mmHg at every step of 40 mmHg and 4 s, and then returned to 0 mmHg in similar steps. ISP and mean arterial pressure (MAP) were recorded simultaneously. ISP-MAP relationship curve was constructed by fitting to the logistic function with five parameters. The CSR parameters and the ISP-MAP relationship curve were separately compared statistically. The main results obtained are as follows. (1) Stress significantly shifted the ISP-MAP relationship curve upwards and obviously moved the middle part of ISP-Gain relationship curve downwards, and decreased the value of the MAP range and maximum gain (G(max)), but increased the threshold pressure (TP), saturation pressure (SP), set point and ISP at G(max) (ISP(Gmax)). (2) I.c.v. of H1 receptor antagonist chlorpheniramine (CHL, 5 microg) or H2 receptor antagonist cimetidine (CIM, 15 g) significantly diminished the above-mentioned changes in CSR performance induced by stress; the alleviative effect of CIM was less remarkable than that of CHL. The responses of CSR in stressed rats to H(1) or H(2) receptor antagonists generally occurred 20 min after the administration and lasted approximately for 15 min. (3) After microinjection of CHL (0.5 microg) or CIM (1.5 microg) into the NTS, the responses of CSR in stressed groups were similar to those after i.c.v. injection of CHL or CIM. (4) However, microinjection of CHL or CIM into the lateral cerebroventricle or the NTS could not completely abolish the stress-induced changes in CSR. These findings suggest that stress results in a resetting of CSR, a decrease in reflex sensitivity. The stress-induced changes in CSR may be mediated, at least in part, by activating the brain histaminergic system. The central histaminergic receptors (H(1) and H(2) receptors) may play an important role in the resetting of CSR under stress. The descending histaminergic pathway from the hypothalamus to NTS may be involved in these effects.

[Effect of intracerebroventricular injection of histamine on carotid sinus baroreceptor reflex in anesthetized rats and its mechanism].

The changes in carotid sinus baroreceptor reflex (CSR) performance induced by intracerebroventricular injection (i.c.v.) of histamine (HA) were investigated. The effects of pretreatment with HA receptors antagonists into the cerebroventricle or nucleus of solitary tract (NTS) on the responses of CSR to HA were also examined. Intracarotid sinus pressure (ISP)-mean arterial pressure (MAP) relationship curve was constructed by fitting to the logistic function with five parameters in 50 Wistar rats anesthetized with pentobarbital sodium. The left and right carotid sinus regions were isolated from the systemic circulation and the ISP was altered in a stepwise manner. The main results obtained are as follows. (1) i.c.v. injection of HA (100 ng) significantly shifted the ISP-MAP relationship curve upwards and moved the middle part of ISP-Gain relationship curve downwards, and reduced the MAP range and maximum gain (G(max)), but increased the threshold pressure (TP), saturation pressure (SP) and ISP at G(max) (ISP (Gmax)). (2) The pretreatment with H(1) or H(2) receptors antagonist, chlorpheniramine (CHL, 5 microg) or cimetidine (CIM, 15 microg), could obviously diminish the above-mentioned changes in CSR performance induced by HA, but the effect of CIM was less remarkable than that of CHL. (3) The pretreatment with both CHL and CIM (5 microg and 15 microg) at the same time abolished the responses of CSR performance to HA completely. (4) After microinjection of CHL (0.5 microg) or CIM (1.5 microg) into the NTS, the responses of CSR to HA were similar to those after i.c.v. CHL or CIM, but the change in TP was significantly decreased. These findings suggest that the intracerebroventricular administration of HA results in a rapid resetting of CSR and a decrease in reflex sensitivity. The response of CSR to HA might be mediated by both central H(1) and H(2) receptors, especially by H(1) receptors. The effects of the central HA on CSR might be related to a histaminergic descending pathway from the hypothalamus to NTS. It is suggested that the HA receptors in the NTS play an important role in the responses of CSR to HA.

Chronic restraint stress inhibits hair growth via substance P mediated by reactive oxygen species in mice.

BACKGROUNDS: Solid evidence has demonstrated that psychoemotional stress induced alteration of hair cycle through neuropeptide substance P (SP) mediated immune response, the role of reactive oxygen species (ROS) in brain-skin-axis regulation system remains unknown. OBJECTIVES: The present study aims to investigate possible mechanisms of ROS in regulation of SP-mast cell signal pathway in chronic restraint stress (CRS, a model of chronic psychoemotional stress) which induced abnormal of hair cycle. METHODS AND RESULTS: Our results have demonstrated that CRS actually altered hair cycle by inhibiting hair follicle growth in vivo, prolonging the telogen stage and delaying subsequent anagen and catagen stage. Up-regulation of SP protein expression in cutaneous peripheral nerve fibers and activation of mast cell were observed accompanied with increase of lipid peroxidation levels and reduction of the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in CRS mice skin. In addition, SP receptor antagonist (RP67580) reduced mast cell activations and lipid peroxidation levels as well as increased GSH-Px activity and normalized hair cycle. Furthermore, antioxidant Tempol (a free radical scavenger) also restored hair cycle, reduced SP protein expression and mast cell activation. CONCLUSIONS: Our study provides the first solid evidence for how ROS play a role in regulation of psychoemotional stress induced SP-Mast cell pathway which may provide a convincing rationale for antioxidant application in clinical treatment with psychological stress induced hair loss.

Apoptosis and autophagy contribute to gender difference in cardiac ischemia-reperfusion induced injury in rats.

AIMS: Gender difference in cardiac ischemia-reperfusion (IR) induced injury has been reported in animal models. However, a large-scale clinical trial found an increase in cardiovascular incidents in women with hormone replacement therapy. The present study is aimed to explore possible mechanisms of gender difference in cardiac IR induced injury. MAIN METHODS: Male and female Sprague-Dawley rats were subjected to a 30-min coronary arterial occlusion followed by reperfusion. The infarct size and apoptotic cell number at 24h after reperfusion were significantly lower in female rats than in male rats. KEY FINDINGS: Male rats expressed higher anti-apoptotic protein Bcl2 levels compared with female rats under physiological conditions. However, levels of Bcl2 were reduced significantly after IR in male rats but not in, female rats. Levels of pro-apoptotic protein, Bax and phospho-p38, showed similar under physiological conditions. In response to IR expression of Bax was markedly reduced in female rats but not in male rats, and expression of phospho-p38 was significantly increased in male rats but not in female rats. In addition, female rats showed marked increase of autophagy marker, ratio of LC3B to LC3A, while male rats significantly decreased the ratio in response to IR. SIGNIFICANCE: Gender difference in IR injury is due to the different regulation of anti-apoptotic protein, pro-apoptotic protein and autophagy protein levels in male rats and levels in female rats. Our results provide better understanding of sex differences in cardiac IR injury.

Cobratoxin inhibits pain-evoked discharge of neurons in thalamic parafascicular nucleus in rats: involvement of cholinergic and serotonergic systems.

The present study investigated the inhibitory effect of cobratoxin (CTX) on pain-evoked discharge of neurons in thalamic parafascicular nucleus (Pf) of rats and analyzed some of the mechanisms involved in this effect. Intracerebroventricular injection (icv) of CTX at 0.56, 1.12 and 4.50 microg/kg resulted in a dose-dependent inhibitory effect on the pain-evoked discharges of Pf neurons. The inhibition of pain-evoked discharges of Pf neurons by CTX at high dose (4.50 microg/kg) persisted at least for 2h, while the inhibitory effect of morphine (40 microg) persisted no longer than 30 min. The inhibitory effect of CTX was reversed by pretreatment with atropine (icv, 5 microg). In contrast, icv injection of naloxone (4 microg) had no effect on CTX-induced inhibition. Furthermore, pretreatment with parachlorophenylalanine, a specific inhibitor of tryptophan hydroxylase, also significantly attenuated the inhibitory effect of CTX. The results suggested that: (a) CTX has a dose-dependent inhibitory effect on pain-evoked discharges of Pf neurons, confirming electrophysiologically the antinociceptive action of CTX; (b) the inhibitory effect of CTX has a longer duration compared to that of morphine; (c) central cholinergic and serotonergic systems, but not opioidergic system, are involved in the inhibitory effect of CTX.

Hemodilution does not alter the coronary vasodilating effects of endogenous or exogenous nitric oxide.

INTRODUCTION: It is well known that hemoglobin is a scavenger of nitric oxide (NO). The present study used a canine model to test the hypothesis that acute normovolemic hemodilution (ANH) affects NO-mediated coronary vasodilation. METHODS: Studies were performed in 18 open-chest, anesthetized dogs. In Series 1, the contribution of endogenous NO to coronary vasodilatation during ANH with 5% dextran-40 (reduction in hematocrit by 50%) was assessed. This was accomplished by comparing myocardial blood flow (MBF; radioactive microspheres) in the left anterior descending (LAD) region, which was treated with the NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), to that in the circumflex (control) region. In Series 2, the LAD was perfused via a controlled-pressure extracorporeal system with coronary blood flow (CBF) measured with an ultrasonic, transit-time flow transducer. The dose-dependent increases in CBF caused by acetylcholine (ACh), which releases endogenous NO from the vascular endothelium, and sodium nitroprusside (SNP), which provides exogenous NO, were compared before and during ANH. RESULTS: Acute normovolemic hemodilution caused similar (approximately twofold) increases in MBF (P < 0.01) in the absence and presence of L-NAME, and it did not affect the dose-related increases in CBF caused by ACh and SNP. CONCLUSIONS: Series 1: under baseline conditions, hemoglobin in red blood cells does not limit the coronary vasodilatation resulting from tonic release of NO; NO does not mediate coronary vasodilation during ANH. Series 2: ANH does not influence the coronary vasodilating effects of increased levels of NO, whether due to endogenous release (ACh) or infusion of an NO donor (SNP).

[A receptors in the NTS modulate depression of carotid baroreflex induced by intracerebroventricular injection of histamine in rats].

AIM: To investigate the roles of alpha1 and alpha2 receptors in the nucleus tractus solitarius (NTS) in the carotid baroreflex (CBR) resetting induced by the intracerebroventricular injection (ICV) of histamine (HA). METHODS: The left and right carotid sinus regions were isolated from the systemic circulation in 25 Sprague-Dawley rats anesthetized with pentobarbital sodium. The intracarotid sinus pressure (ISP) was altered in a stepwise manner. ISP-mean arterial pressure (MAP) relationship curve and its characteristic parameters were constructed by fitting to the logistic function with five parameters. The changes in CBR performance induced by ICV HA and the effects of pretreatment with alpha1 or alpha2 receptor antagonist into the NTS on the responses of CBR to HA were examined. RESULTS: ICV HA (60 micromol x L(-1) in 5 microl) significantly shifted the ISP-MAP relationship curve upwards (P < 0.05) and moved the middle part of ISP-Gain relationship curve downwards (P < 0.05), and reduced the MAP range and maximum gain (P < 0.05). The pretreatment with phenoxybenzamine (PBZ, a selective antagonist of alpha1 receptor, 3 micromol x L(-1) in 500 nl) or yohimbine (YOH, a selective antagonist of alpha2 receptor, 2.5 micromol x L(-1) in 500 nl) into the NTS could obviously intensify the above-mentioned changes in CBR performance induced by HA, but the intensive effect of PBZ was less remarkable than that of YOH (P < 0.05). CONCLUSION: The intracerebroventricular administration of HA results in a rapid resetting of CBR and a decrease in reflex sensitivity, and the functions of alpha1 and alpha2 receptors in the NTS might weaken CBR resetting induced by ICV HA. Furthermore, alpha2 receptor in the NTS might play an more important role in modulating the responses of CHR to HA.