[The value of color doppler ultrasonography in the diagnosis of endograft infections following endovascular aneurysm repair].

Objective: To study the value of color doppler ultrasonography (CDU) in the diagnosis of endograft infections following endovascular aneurysm repair (EVAR). Methods: The retrospective analysis of post-EVAR stent infections identified by computed tomography angiography (CTA) was conducted at the First Affiliated Hospital of Sun Yat-sen University from January 2010 to December 2020. There were 16 males and 4 females, aged from 49 to 86 years. All the patients were detected by CDU. The endoleak, bubbles, abscess, hematoma, aortic intestinal fistula (AEF) and occlusion of stent detected by CTA and CDU were analyzed and compared. Results: Among 20 patients, 9 cases with endoleak were detected by CTA, while CDU showed 8 cases with endoleak. The rate of missed diagnosis was 1/9. The misdiagnosis rate was 0, and the Youden index was 0.89. CDU detected 3 cases with type Ⅱ endoleak, and 1 case was missed when compared with CTA. Three cases with type Ⅰa and 2 cases with type Ⅰb were detected by CDU, which were consistent with those of CTA. CDU and CTA showed that there were no cases with type Ⅲ and type Ⅳ endoleaks. CDU detected 8 cases with bubbles in the sac. Compared with CTA, the rate of missed diagnosis was 2/10. The misdiagnosis rate was 0, and the Youden index was 0.80. The cases with abscess, hematoma, increasing size of the aneurysm, occlusion of stent and fluid sonolucent area in the sac detected by CDU were 8/20, 2/20, 4/20, 1/20, 2/20, which were consistent with CTA. CDU did not detect the 3 cases with aortoenteric fistula(AEF) which were identified by CTA. The follow-up of CDU showed that the extra-anatomic bypasses remained their patency in 5 cases, 1 case occurred bypass occlusion. The range of infectious area and bubbles reduced in 2 cases. There was no change of endoleak in 1 case. All the follow-up results were consistent with those of CTA. Conclusion: CDU can comprehensively evaluate the infection in and around the aneurysm in patients with stentinfection after EVAR, with a high auuraly, and has important clinical significance for the early diagnosis, treatment and prognosis of patients.

[Efficacy of endovenous microwave ablation in treating primary varicose veins of the lower extremities].

Objective: To explore the efficacy of endovenous microwave ablation in treating primary varicose veins of the lower extremities. Methods: A total of 214 patients with primary varicose veins of the lower extremities in the First Affiliated Hospital of Sun Yat-sen University from January 2017 to November 2019 were included and divided into microwave ablation group (n=128) and high ligation with microwave ablation group (n=86) according to surgical approach. Further subgroups including subgroup S (diameter<10 mm) and subgroup L (diameter≥10 mm) were established. The surgical success rate, postoperative incidence and recovery of local skin numbness and ecchymosis, and the postoperative recovery of active skin ulcer were compared between the two groups and subgroups. Results: The surgical success rate was 96% (136/141) in microwave ablation group, 100% in subgroup S (116/116), and 80% in subgroup L (20/25), respectively. In addition, the surgical success rate in high ligation with microwave ablation group, subgroup S, and subgroup L was all 100% (90/90, 73/73, and 17/17). In subgroup L of microwave ablation group, the diameter of 5 great saphenous veins in patients who failed the microwave ablation was 13.0-17.1 mm. The mean follow-up time was (24±4) months in microwave ablation group and (36±6) months in high ligation with microwave ablation group, respectively. In the microwave ablation group and high ligation with microwave ablation group, the incidence of postoperative skin numbness was 15.6% and 14.4%, respectively, and the incidence of skin ecchymosis was 5.7% and 3.3%, respectively, with no statistically significant difference between the two groups (both P>0.05). The rate of active skin ulcer in the two groups was 6.4% and 15.6%, respectively, and the difference was statistically significant (P=0.020). Local skin ecchymosis in the two groups recovered within 1 month after operation. Local skin numbness in both groups recovered within the maximum 2 years of follow-up, and active skin ulcer in both groups recovered within the maximum 1 years of follow-up. Conclusion: The endovenous microwave ablation is safe and effective, especially combining with high ligation of great saphenous vein. Good follow-up results can be achieved for great saphenous vein with diameter smaller than 10 mm. However, for those with diameter greater than 10 mm, the surgical success rate of endovenous microwave ablation decreases.

CLIC1 knockout inhibits invasion and migration of gastric cancer by upregulating AMOT-p130 expression.

PURPOSE: To explore the regulatory relationship between Chloride intracellular channel 1 (CLIC1) and Angiomotin (AMOT)-p130, and reveal the role of AMOT-p130 in gastric cancer (GC). METHODS: Immunohistochemistry was performed to analyze the expression of CLIC1 and AMOT-p130 in GC tissues and adjacent tissues. The expression of AMOT-p130 upon CLIC1 silencing was analyzed using RT-PCR, western blot, and immunofluorescence in GC cells. Transwell and wound-healing assays were performed to detect migration and invasion in GC cells. The changes in EMT-related proteins were detected using western blot. RESULTS: Our study found that high CLIC1 expression was significantly associated with low AMOT-p130 expression in GC tissues. Silencing CLIC1 expression in MGC-803 cells (MGC-803 CLIC1 KO) and AGS cells (AGS CLIC1 KO) decreased the invasive and migratory abilities of tumor cells, which were induced by the upregulation of AMOT-p130. Subsequently, we demonstrated that AMOT-p130 inhibits the invasive and migratory abilities of GC cells by inhibiting epithelial-mesenchymal transition. CONCLUSIONS: Our study suggests that AMOT-p130 could inhibit epithelial-mesenchymal transition in GC cells. CLIC1 may participate in the metastatic progression of GC by downregulating the expression of AMOT-p130.

[Mid- and long-term results of endovascular aneurysm repair for abdominal aortic aneurysms: a single center's experience for 10 years].

Objective: To examine the mid- and long-term outcomes of endovascular aneurysm repair (EVAR). Methods: This was a retrospective cohort study of 540 patients with abdominal aortic aneurysm who received EVAR at Department of Vascular Surgery, The First Affiliated Hospital, Sun Yat-sen University from January 2009 to December 2018. There were 503 males and 37 females, aged of (69±8) years (range: 44 to 87 years). Clinical data including concomitant disease, aneurysm size and surgical data were collected and patients were followed up after operation. The cumulative survival rate was assessed using the Kaplan-Meier estimator and multivariate Cox regression was used to analyze the independent prognosis factors. Results: The technical success rate was 91.1% (492/540). The perioperative mortality rate was 1.3% (7/540) and the follow-up rate was 91.7% (489/533). The median follow-up time was 45(63) months (range: 1 to 133 months). The all-cause mortality rate was 21.3% (104/489) and the aneurysm-related mortality rate was 6.3% (31/489) during follow-up period. The overall cumulative survival rate of 1-, 3-, 5- and 10-year were 95.1%, 84.0%, 69.5% and 38.6%, respectively, while freedom from aneurysm-related death were 98.4%, 93.3%, 88.4% and 84.4%. During the follow-up period, the complications rate was 9.0% (44/489), and the re-intervention rate was 4.9% (24/489). Cox regression analysis showed that elder age (HR=2.15, 95%CI: 1.41 to 3.26, P<0.01), preoperative aneurysm rupture (HR=2.72, 95%CI: 1.78 to 4.15, P<0.01) and short neck aneurysm (HR=1.97, 95%CI: 1.07 to 3.61, P=0.029) were independent prognosis factors for long-term survival after EVAR. Connclusion: EVAR has low perioperative mortality, high technical success rate, and satisfactory mid-and long-term outcomes.

[The value of color doppler ultrasonography in the diagnosis of impending ruptured abdominal aortic aneurysm].

Objective: To study the value of color doppler ultrasonography (CDU) in diagnosis of impending ruptured abdominal aortic aneurysm (IRAAA). Methods: A total of 35 cases with IRAAA which were identified by CDU in our department from June 2014 to June 2019 were retrospectively analyzed. All the patients were detected by computed tomographic angiography (CTA). The types, length of the neck of aneurysm, largest diameter, thrombosis, involvement of common iliac artery and impending ruptured conditions were compared. The postoperative patients were followed-up by CDU and CTA (mean time was 2.6 months). Results: Among 35 patients, CDU diagnosed that 5 cases were pararenal types and 30 cases were infrarenal types. CTA showed that 4 cases were pararenal types and 31 cases were infrarenal types. The misdiagnosis rate of CDU was 2.9% (1/35). CDU showed that bilateral common iliac arteries were involved in 21 cases, right common iliac arteries were involved in 3 cases, and left common iliac arteries were involved in 2 cases. CTA detected the same results. There was no statistical difference between CDU and CTA for detection of the largest anteroposterior diameter, transverse diameter and the thickness of thrombosis (P values were 0.354, 0.310 and 0.865). There was statistical difference in the detection of the length of the aneurysm's neck (P=0.006). CDU showed 3 cases of focal wall discontinuity, 4 cases of hyperattenuating crescent sign, 3 cases of thrombus fissuration and 2 cases of saclike protuberance, which were consistent with the detection of CTA. CDU showed that locally thin wall of aneurysm was detected in the rest of 23 cases. CTA showed 2 patients were negative. The misdiagnosis rate of CDU was 5.7% (2/35). Three cases did not undergo surgery. Open repairs (OR) were performed in 5 cases. Endovascular aneurysm repairs (EVAR) were performed in 27 cases. The postoperative patients were followed up by CDU and CTA at 1 month, 3 months, 6 months and 12 months. All the artificial blood vessels and stents were patent. Endoleak was observed in 5 cases. Aneurysm sac thrombosis was found in the rest of the cases. Conclusions: CDU plays an important role in the analysis and diagnosis of the size, range, and the impending ruptured symptoms of the aneurysm. It provides a reliable basis for IRAAA screening, diagnosis and postoperative follow-up, and has important clinical value.

[Risk factors of rupture of internal carotid artery during surgical resection of carotid body tumor].

Objective: To investigate risk factors of rupture of internal carotid artery resection during carotid body tumor resection and to summarize our treatment experience. Methods: During the period from 1991 to 2016, rupture of internal carotid artery occurred in 27 patients (28 tumors) during surgical resection of carotid body tumor in the First Affiliated Hospital of Sun Yat-sen University. Their clinical and follow-up data were retrospectively collected and analyzed. For all patients underwent surgical resection during this period, Logistic regression analysis was used to investigate the risk factors of intraoperative rupture of internal carotid artery. Results: Of these 28 tumors, there were 15 (53.6%) tumors with diameter≥5 cm and 20 (71.4%) Shamblin Ⅲ tumors. Intraoperatively, shunt was applied for 8 (28.6%) cases. Thirteen (46.4%) patients underwent ligation of external carotid artery, while 2 (7.1%) patients accepted resection of cranial nerves. Direct closure/patchplasty, autologous vessels or graft reconstruction was used in 16, 10 and 2 cases, respectively. Postoperatively, stroke occurred in 4(14.3%) cases and cranial nerve deficit in 15 (53.6%) cases. During a median length of 36 (14-125) months, cranial nerve deficit persisted in 5 cases. Follow-up radiologic examination indicated 3 (10.7%) cases of targeted vessel occlusion. However, no new-onset stroke was identified. Among all patients underwent surgical resection of carotid body tumor, female (OR=3.650, P=0.012), age≤25 years old (OR=3.710, P=0.013) and Shamblin Ⅲ tumor (OR=4.631, P=0.008) increase the risks of intraoperative carotid artery rupture. Conclusions: Shamblin Ⅲ tumor is the predictor of rupture of internal carotid artery. Intraoperative, properly increased blood pressure, intraoperative heparinization and use of shunt for those cases without well-compensated cranial collateral arteries are likely to decreasing the incidence of stroke.

[Management of endo-graft infection following endovascular repair of abdominal aortic aneurysm].

OBJECTIVE: To summarize our experience in the management of stent-graft infection after endo-vascular aortic repair (EVAR) of abdominal aortic aneurysm (AAA). METHODS: Data of patients who were diagnosed as endo-graft infection following EVAR and admitted in our center between January 2000 and December 2015 were reviewed. Clinical records including causes of infection, medical history, re-operative procedures, and prognostic data were analysed. RESULTS: A total of 10 male patients, aged 45-72 years (averaged 62.5 years), were enrolled. Two patients received previous EVAR in our center, accounting for 0.23% of all the 885 EVAR procedures we conducted during the same period. The symptoms related to stent infection, including recurrent fever (100%) and persistent back pain (40%), occured 0 to 27 months (averaged 6.9 months) after EVAR procedure. Eight patients were found to have apparent causes (80%), including 1 case with upper respiratory infection and sepsis, 4 cases with aorto-enteric fistula (AEF) and 3 cases with inflammatory AAA. Except one DNR, other 9 patients received re-operation, including 1 procedure of open debridement and drainage, 1 procedure of endo-graft excision and bilateral axillary-femoral bypass, 7 procedures of endograft excision and axillary-bifemoral bypass. During the follow-up period(2-60 months, averaged 24.1 months), 1 patient was lost, 1 patient died from aortic stump rupture (12.5%) and other 7 patients survived. Bypass occlusion was observed in 1 patient (12.5%) without severe limb ischemia. CONCLUSIONS: AEF and inflammatory AAA are two leading causes of endo-graft infection following EVAR in our patients. Graft excision and axillofemoral bypass is an acceptable management for this life-threatening morbidity.

Prenatal fat exposure and hypothalamic PPAR ß/δ: Possible relationship to increased neurogenesis of orexigenic peptide neurons.

Gestational exposure to a fat-rich diet, while elevating maternal circulating fatty acids, increases in the offspring's hypothalamus and amygdala the proliferation and density of neurons that express neuropeptides known to stimulate consummatory behavior. To understand the relationship between these phenomena, this study examined in the brain of postnatal offspring (day 15) the effect of prenatal fat exposure on the transcription factor, peroxisome proliferator-activated receptor (PPAR) ß/δ, which is sensitive to fatty acids, and the relationship of PPAR ß/δ to the orexigenic neuropeptides, orexin, melanin-concentrating hormone, and enkephalin. Prenatal exposure to a fat-rich diet compared to low-fat chow increased the density of cells immunoreactive for PPAR ß/δ in the hypothalamic paraventricular nucleus (PVN), perifornical lateral hypothalamus (PFLH), and central nucleus of the amygdala (CeA), but not the hypothalamic arcuate nucleus or basolateral amygdaloid nucleus. It also increased co-labeling of PPAR ß/δ with the cell proliferation marker, BrdU, or neuronal marker, NeuN, and the triple labeling of PPAR ß/δ with BrdU plus NeuN, indicating an increase in proliferation and density of new PPAR ß/δ neurons. Prenatal fat exposure stimulated the double-labeling of PPAR ß/δ with orexin or melanin-concentrating hormone in the PFLH and enkephalin in the PVN and CeA and also triple-labeling of PPAR ß/δ with BrdU and these neuropeptides, indicating that dietary fat increases the genesis of PPAR ß/δ neurons that produce these peptides. These findings demonstrate a close anatomical relationship between PPAR ß/δ and the increased proliferation and density of peptide-expressing neurons in the hypothalamus and amygdala of fat-exposed offspring.

Effects of embryonic ethanol exposure at low doses on neuronal development, voluntary ethanol consumption and related behaviors in larval and adult zebrafish: Role of hypothalamic orexigenic peptides.

Embryonic exposure to ethanol is known to affect neurochemical systems in rodents and increase alcohol drinking and related behaviors in humans and rodents. With zebrafish emerging as a powerful tool for uncovering neural mechanisms of numerous diseases and exhibiting similarities to rodents, the present report building on our rat studies examined in zebrafish the effects of embryonic ethanol exposure on hypothalamic neurogenesis, expression of orexigenic neuropeptides, and voluntary ethanol consumption and locomotor behaviors in larval and adult zebrafish, and also effects of central neuropeptide injections on these behaviors affected by ethanol. At 24h post-fertilization, zebrafish embryos were exposed for 2h to ethanol, at low concentrations of 0.25% and 0.5%, in the tank water. Embryonic ethanol compared to control dose-dependently increased hypothalamic neurogenesis and the proliferation and expression of the orexigenic peptides, galanin (GAL) and orexin (OX), in the anterior hypothalamus. These changes in hypothalamic peptide neurons were accompanied by an increase in voluntary consumption of 10% ethanol-gelatin and in novelty-induced locomotor and exploratory behavior in adult zebrafish and locomotor activity in larvae. After intracerebroventricular injection, these peptides compared to vehicle had specific effects on these behaviors altered by ethanol, with GAL stimulating consumption of 10% ethanol-gelatin more than plain gelatin food and OX stimulating novelty-induced locomotor behavior while increasing intake of food and ethanol equally. These results, similar to those obtained in rats, suggest that the ethanol-induced increase in genesis and expression of these hypothalamic peptide neurons contribute to the behavioral changes induced by embryonic exposure to ethanol.

Prenatal exposure to ethanol stimulates hypothalamic CCR2 chemokine receptor system: Possible relation to increased density of orexigenic peptide neurons and ethanol drinking in adolescent offspring.

Clinical and animal studies indicate that maternal consumption of ethanol during pregnancy increases alcohol drinking in the offspring. Possible underlying mechanisms may involve orexigenic peptides, which are stimulated by prenatal ethanol exposure and themselves promote drinking. Building on evidence that ethanol stimulates neuroimmune factors such as the chemokine CCL2 that in adult rats is shown to colocalize with the orexigenic peptide, melanin-concentrating hormone (MCH) in the lateral hypothalamus (LH), the present study sought to investigate the possibility that CCL2 or its receptor CCR2 in LH is stimulated by prenatal ethanol exposure, perhaps specifically within MCH neurons. Our paradigm of intraoral administration of ethanol to pregnant rats, at low-to-moderate doses (1 or 3g/kg/day) during peak hypothalamic neurogenesis, caused in adolescent male offspring twofold increase in drinking of and preference for ethanol and reinstatement of ethanol drinking in a two-bottle choice paradigm under an intermittent access schedule. This effect of prenatal ethanol exposure was associated with an increased expression of MCH and density of MCH(+) neurons in LH of preadolescent offspring. Whereas CCL2(+) cells at this age were low in density and unaffected by ethanol, CCR2(+) cells were dense in LH and increased by prenatal ethanol, with a large percentage (83-87%) identified as neurons and found to colocalize MCH. Prenatal ethanol also stimulated the genesis of CCR2(+) and MCH(+) neurons in the embryo, which co-labeled the proliferation marker, BrdU. Ethanol also increased the genesis and density of neurons that co-expressed CCR2 and MCH in LH, with triple-labeled CCR2(+)/MCH(+)/BrdU(+) neurons that were absent in control rats accounting for 35% of newly generated neurons in ethanol-exposed rats. With both the chemokine and MCH systems believed to promote ethanol consumption, this greater density of CCR2(+)/MCH(+) neurons in the LH of preadolescent rats suggests that these systems function together in promoting alcohol drinking during adolescence.

Model of voluntary ethanol intake in zebrafish: effect on behavior and hypothalamic orexigenic peptides.

Recent studies in zebrafish have shown that exposure to ethanol in tank water affects various behaviors, including locomotion, anxiety and aggression, and produces changes in brain neurotransmitters, such as serotonin and dopamine. Building on these investigations, the present study had two goals: first, to develop a method for inducing voluntary ethanol intake in individual zebrafish, which can be used as a model in future studies to examine how this behavior is affected by various manipulations, and second, to characterize the effects of this ethanol intake on different behaviors and the expression of hypothalamic orexigenic peptides, galanin (GAL) and orexin (OX), which are known in rodents to stimulate consumption of ethanol and alter behaviors associated with alcohol abuse. Thus, we first developed a new model of voluntary intake of ethanol in fish by presenting this ethanol mixed with gelatin, which they readily consume. Using this model, we found that individual zebrafish can be trained in a short period to consume stable levels of 10% or 20% ethanol (v/v) mixed with gelatin and that their intake of this ethanol-gelatin mixture leads to pharmacologically relevant blood ethanol concentrations which are strongly, positively correlated with the amount ingested. Intake of this ethanol-gelatin mixture increased locomotion, reduced anxiety, and stimulated aggressive behavior, while increasing expression of GAL and OX in specific hypothalamic areas. These findings, confirming results in rats, provide a method in zebrafish for investigating with forward genetics and pharmacological techniques the role of different brain mechanisms in controlling ethanol intake.

Common effects of fat, ethanol, and nicotine on enkephalin in discrete areas of the brain.

Fat, ethanol, and nicotine share a number of properties, including their ability to reinforce behavior and produce overconsumption. To test whether these substances act similarly on the same neuronal populations in specific brain areas mediating these behaviors, we administered the substances short-term, using the same methods and within the same experiment, and measured their effects, in areas of the hypothalamus (HYPO), amygdala (AMYG), and nucleus accumbens (NAc), on mRNA levels of the opioid peptide, enkephalin (ENK), using in situ hybridization and on c-Fos immunoreactivity (ir) to indicate neuronal activity, using immunofluorescence histochemistry. In addition, we examined for comparison another reinforcing substance, sucrose, and also took measurements of stress-related behaviors and circulating corticosterone (CORT) and triglycerides (TG), to determine if they contribute to these substances' behavioral and physiological effects. Adult Sprague-Dawley rats were gavaged three times daily over 5 days with 3.5 mL of water, Intralipid (20% v/v), ethanol (12% v/v), nicotine (0.01% w/v) or sucrose (22% w/v) (approximately 7 kcal/dose), and tail vein blood was collected for measurements of circulating CORT and TG. On day five, animals were sacrificed, brains removed, and the HYPO, AMYG, and NAc processed for single- or double-labeling of ENK mRNA and c-Fos-ir. Fat, ethanol, and nicotine, but not sucrose, increased the single- and double-labeling of ENK and c-Fos-ir in precisely the same brain areas, the middle parvocellular but not lateral area of the paraventricular nucleus, central but not basolateral nucleus of the AMYG, and core but not shell of the NAc. While having little effect on stress-related behaviors or CORT levels, fat, ethanol, and nicotine all increased circulating levels of TG. These findings suggest that the overconsumption of these three substances and their potential for abuse are mediated by the same populations of ENK-expressing neurons in specific nuclei of the hypothalamus and limbic system.

Prenatal ethanol exposure stimulates neurogenesis in hypothalamic and limbic peptide systems: possible mechanism for offspring ethanol overconsumption.

Exposure to ethanol during the prenatal period contributes to increased alcohol consumption and preference in rodents and increased risk for alcoholism in humans. With studies in adult animals showing the orexigenic peptides, enkephalin (ENK), galanin (GAL) and orexin (OX), to stimulate ethanol consumption, the question addressed here is whether prenatal ethanol alters the development in utero of specific neurons that express these peptides. With reports describing suppressive effects of high doses of ethanol, we examined the offspring of dams gavaged from embryonic day 9 to parturition with a control solution or lower ethanol doses, 1 and 3g/kg/day, known to promote ethanol consumption in the offspring. To understand underlying mechanisms, measurements were taken in postnatal offspring of the expression of ENK in the hypothalamic paraventricular nucleus (PVN) and nucleus accumbens (NAc), GAL in the PVN, and OX in the perifornical lateral hypothalamus (PFLH) using real-time qPCR and in situ hybridization, and also of the cell proliferation marker, 5-bromo-2-deoxyuridine (BrdU), and its double-labeling with either neuronal nuclei (NeuN), a marker of mature neurons, or the peptides. On postnatal day 15 (P15), after two weeks without ethanol, the offspring showed increased expression of ENK in the PVN and NAc core but not shell, GAL in the PVN, and OX in the PFLH. In these same areas, prenatal ethanol compared to control increased the density at birth (P0) of neurons expressing these peptides and at P0 and P15 of neurons double-labeling BrdU and NeuN, indicating increased neurogenesis. These BrdU-positive neurons were found to express ENK, GAL and OX, indicating that prenatal ethanol promotes neurogenesis in these specific peptide systems. There were no changes in gliogenesis or apoptosis. This increase in neurogenesis and density of peptide-expressing neurons suggests the involvement of these hypothalamic and accumbal peptide systems in mediating the increased alcohol consumption observed in prenatal ethanol-exposed offspring.

Galanin and the orexin 2 receptor as possible regulators of enkephalin in the paraventricular nucleus of the hypothalamus: relation to dietary fat.

Recent studies show that the non-opioid peptides, galanin (GAL) and orexin (OX), are similar to the opioid enkephalin (ENK) in being stimulated by dietary fat and also in enhancing the consumption of a high-fat diet (HFD). This suggests that, when an HFD is provided, these non-opioids may stimulate the opioid system to promote excess consumption of this diet. Using single- and double-labeling immunohistochemistry, the present study sought to identify possible neuroanatomical substrates for this close relationship. Focusing on the hypothalamic paraventricular nucleus (PVN), and particularly its anterior (aPVN), middle (mPVN) and posterior (pPVN) parts, the experiments examined whether GAL itself or the receptors for GAL and OX are stimulated by an HFD in the same areas and possibly the same neurons as ENK. Compared to animals fed a standard chow diet, rats consuming an HFD exhibited an increased density of medial parvocellular neurons immunoreactive (IR) for GAL in the mPVN and aPVN and for ENK in the mPVN and pPVN, distinguishing the mPVN as an area where both peptides were affected. While showing little evidence for GAL and ENK colocalization with a chow diet, double-labeling studies in HFD-fed rats revealed significant colocalization specifically in medial parvocellular neurons of the mPVN. Immediately posterior to this site, further analyses revealed a similar relationship between the OX 2 receptor (OX(2)R) and ENK in HFD-treated animals. While increasing the density of neurons immunoreactive for OX(2)R as well as for the GAL 1 receptor but not OX 1 receptor, HFD consumption increased the colocalization only of OX(2)R and ENK, specifically in the medial parvocellular neurons of the pPVN. These changes in HFD-fed rats, showing GAL and OX(2)R to colocalize with ENK exclusively in neurons of the medial parvocellular mPVN and pPVN, respectively, suggest possible neural substrates through which the non-opioid peptides may functionally interact with ENK when exposed to an HFD.

Role of melanin-concentrating hormone in the control of ethanol consumption: Region-specific effects revealed by expression and injection studies.

The peptide melanin-concentrating hormone (MCH), produced mainly by cells in the lateral hypothalamus (LH), perifornical area (PF) and zona incerta (ZI), is suggested to have a role in the consumption of rewarding substances, such as ethanol, sucrose and palatable food. However, there is limited information on the specific brain sites where MCH acts to stimulate intake of these rewarding substances and on the feedback effects that their consumption has on the expression of endogenous MCH. The current study investigated MCH in relation to ethanol consumption, in Sprague-Dawley rats. In Experiment 1, chronic consumption of ethanol (from 0.70 to 2.7 g/kg/day) dose-dependently reduced MCH gene expression in the LH. In Experiments 2-4, the opposite effect was observed with acute oral ethanol, which stimulated MCH expression specifically in the LH but not the ZI. In Experiment 5, the effect of MCH injection in brain-cannulated rats on ethanol consumption was examined. Compared to saline, MCH injected in the paraventricular nucleus (PVN) and nucleus accumbens (NAc) selectively stimulated ethanol consumption without affecting food or water intake. In contrast, it reduced ethanol intake when administered into the LH, while having no effect in the ZI. These results demonstrate that voluntary, chronic consumption of ethanol leads to local negative feedback control of MCH expression in the LH. However, with a brief exposure, ethanol stimulates MCH-expressing neurons in this region, which through projections to the feeding-related PVN and reward-related NAc can promote further drinking behavior.

Increased enkephalin in brain of rats prone to overconsuming a fat-rich diet.

Recent studies have shown that the opioid enkephalin (ENK), acting in part through the hypothalamic paraventricular nucleus (PVN), can stimulate consumption of a high-fat diet. The objective of the present study was to examine sub-populations of Sprague-Dawley rats naturally prone to overconsuming a high-fat diet and determine whether endogenous ENK, in different brain regions, is altered in these animals and possibly contributes to their behavioral phenotype. An animal model, involving a measure of initial high-fat diet intake during a few days of access that predicts long-term intake, was designed to classify rats at normal weight that are either high-fat consumers (HFC), which ingest 35% more calories of the high-fat than low-fat chow diet, or controls, which consume similar calories of these two diets. Immediately after their initial access to the diet, the HFC compared to control rats exhibited significantly greater expression of ENK mRNA, in the PVN, nucleus accumbens and central nucleus of the amygdala, but not the arcuate nucleus or basolateral amygdala. This site-specific increase in ENK persisted even when the HFC rats were maintained on a chow diet, suggesting that it reflects an inherent characteristic that can be expressed independently of the diet. It was also accompanied by a greater responsiveness of the HFC rats to the stimulatory effect of a PVN-injected, ENK analogue, D-ala2-met-enkephalinamide, compared to saline on consumption of the high-fat diet. Thus, normal-weight rats predicted to overconsume a fat-rich diet exhibit disturbances in endogenous ENK expression and functioning that may contribute to their long-term, behavioral phenotype.

Annexin A1 expression and its prognostic significance in human breast cancer.

Annexin A1 (ANXA1) is a calcium- and phospholipid-binding protein and is considered to play an important role in tumorigenesis. However, the correlation between ANXA1 expression and tumor clinicopathological features in patients with breast cancer remains unclear. This study investigated the prognostic value of ANXA1 protein as breast cancer marker. Tissue microarray blocks, containing 20 cases of non-tumor breast tissue, 20 cases of benign breast lesion and 135 cases of breast cancer (107 with lymph node metastasis), were constructed. Expression of ANXA1 in these specimens was analyzed using immunohistochemistry. In non-tumor tissue and benign breast lesions, myoepithelial cells showed strong expression of ANXA1. Negative ANXA1 expression was significantly associated with advanced disease stage (P<0.05), especially pathological-N stage (P<0.01). The patients with loss of ANXA1 expression in tumor tissues showed a significantly worse overall survival compared with positive ones (P<0.05). ANXA1 did not correlate well with estrogen receptor (ER), progesterone receptor (PR) and HER2/neu status. Moreover, the level of ANXA1 expression in lymph node metastases was higher than corresponding primary breast cancer. These results suggest that ANXA1 may play a multifaceted role in breast cancer development, progression, and metastases.

Circulating triglycerides after a high-fat meal: predictor of increased caloric intake, orexigenic peptide expression, and dietary obesity.

Recent studies in normal-weight rats have linked circulating triglycerides (TG), when elevated by a high-fat (HF) compared to equicaloric low-fat (LF) meal, to an increase in subsequent food intake and hypothalamic expression of orexigenic peptides. The present study tested whether natural variations between rats in their TG levels after a small HF meal can also be related to their individual patterns of eating and peptide expression. In tail vein blood collected on three separate days 60 min after a HF meal, levels of TG were found to be strongly, positively correlated within rats from day to day but were highly variable between rats (75-365 mg/dl), allowing distinct subgroups (33% lowest or highest) to be formed. Compared to "Low-TG responders" with post-meal levels averaging 109 mg/dl, "High-TG responders" with 240 mg/dl showed in two separate experiments a significant increase in caloric intake in a subsequent laboratory chow meal. Before this larger meal, these rats with elevated TG consistently exhibited higher expression levels and synthesis of the orexigenic peptides, enkephalin, orexin and melanin-concentrating hormone, as revealed using real-time quantitative PCR, radiolabeled in situ hybridization, and immunofluorescence histochemistry. Over the long-term, the High-TG responders also showed an increased propensity to overeat, gain weight and accumulate excess body fat on a chronic HF diet. This simple measure of TG levels after a HF meal may offer a useful tool for identifying subpopulations with increased risk for overeating and dietary obesity and detecting early signs of brain disturbances that may contribute to this high-risk phenotype.

Puberty onset in female rats: relationship with fat intake, ovarian steroids and the peptides, galanin and enkephalin, in the paraventricular and medial preoptic nuclei.

Puberty is a time of rapid change, including a marked increase in fat consumption and body fat accrual, particularly in females. The mechanisms underlying these changes are unknown. Building on the results obtained in adult rats, the present study in pubertal rats focused on the orexigenic peptides, galanin (GAL) and enkephalin (ENK), in the paraventricular nucleus (PVN) and medial preoptic nucleus (MPN), which are known to be responsive to female steroids and have a role in both energy balance and reproductive function. The present study examined female rats maintained on pure macronutrient diets from before weaning (day 15) to day 70. After an initial burst in protein intake (days 21-35), rats showed an increase, specifically in preference for fat, from 15% to 30%. In rats examined at different ages before (day 30) and after (days 45 and 60) puberty, this rise in fat intake was associated with a marked increase, from days 30-45, in levels of oestradiol and progesterone and in GAL and ENK mRNA or peptide levels, specifically in the PVN and MPN, but not other hypothalamic areas examined. This positive relationship with increased fat intake, steroids and peptides across ages was also observed when comparing pubertal rats that naturally preferred fat (> 25% of total diet) with those consuming little fat (< 15%) or rats that reached puberty at an early age (days 30-34) with those that were late (days 37-40). These rats with early puberty onset exhibited a strong fat preference 3-4 days before vaginal opening, which was positively related to steroid levels, GAL, fat intake and body fat accrual after puberty. These findings suggest that, in addition to providing a signal for puberty onset, early fat ingestion acting through mechanisms involving the steroids and orexigenic peptides may be related to long-term patterns of eating and body weight regulation.