[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.

[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.

[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.

[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.

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.

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.

Apoptosis: final common pathway of photoreceptor death in rd, rds, and rhodopsin mutant mice.

Mutations in the retinal degeneration, retinal degeneration slow(/peripherin) and rhodopsin genes cause photoreceptor degeneration in humans and mice. Although the phenotypes arising from these mutations are different, suggesting different mechanisms of pathogenesis, we present evidence that apoptosis may be the final common pathway of the disease process linking genotype to phenotype. We observed internucleosomal cleavage of retinal DNA by gel electrophoresis and fragmented DNA at the single cell level by labeling the nicked DNA ends with biotinylated poly(dU). In retinal degeneration mice, DNA fragmentation occurred during the period of photoreceptor degeneration. In retinal degeneration slow mice and in transgenic mice expressing a mutant (Pro347Ser) rhodopsin gene, DNA fragmentation occurred after normal histogenetic cell death (also apoptosis) had ceased. Since DNA fragmentation by internucleosomal cleavage is a cardinal feature of apoptosis, our data suggest that all three of these genetic mutations lead to apoptosis.

Increased caloric intake on a fat-rich diet: role of ovarian steroids and galanin in the medial preoptic and paraventricular nuclei and anterior pituitary of female rats.

Previous studies in male rats have demonstrated that the orexigenic peptide galanin (GAL), in neurones of the anterior parvocellular region of the paraventricular nucleus (aPVN) projecting to the median eminence (ME), is stimulated by consumption of a high-fat diet and may have a role in the hyperphagia induced by fat. In addition to confirming this relationship in female rats and distinguishing the aPVN-ME from other hypothalamic areas, the present study identified two additional extra-hypothalamic sites where GAL is stimulated by dietary fat in females but not males. These sites were the medial preoptic nucleus (MPN), located immediately rostral to the aPVN, and the anterior pituitary (AP). The involvement of ovarian steroids, oestradiol (E(2)) and progesterone (PROG), in this phenomenon was suggested by an observed increase in circulating levels of these hormones and GAL in MPN and AP with fat consumption and an attenuation of this effect on GAL in ovariectomised (OVX) rats. Furthermore, in the same four areas affected by dietary fat, levels of GAL mRNA and peptide immunoreactivity were stimulated by E(2) and further by PROG replacement in E(2)-primed OVX rats and were higher in females compared to males. Because both GAL and PROG stimulate feeding, their increase on a fat-rich diet may have functional consequences in females, possibly contributing to the increased caloric intake induced by dietary fat. This is supported by the findings that PROG administration in E(2)-primed OVX rats reverses the inhibitory effect of E(2) on total caloric intake while increasing voluntary fat ingestion, and that female rats with higher GAL exhibit increased preference for fat compared to males. Thus, ovarian steroids may function together with GAL in a neurocircuit, involving the MPN, aPVN, ME and AP, which coordinate feeding behaviour with reproductive function to promote consumption of a fat-rich diet at times of increased energy demand.

Increased caloric intake after a high-fat preload: relation to circulating triglycerides and orexigenic peptides.

To investigate mechanisms that mediate the greater food intake induced by a fat-rich diet, the present study tested an acute "preload-to-test meal" paradigm in normal-weight rats. In this paradigm, the rats were given a small high-fat (HF) compared to low-fat (LF) preload and, after an intermeal interval, allowed to consume freely on a subsequent test meal. Modified versions of this paradigm were tested to determine the robustness of the greater caloric intake induced by the HF preload while standardizing the test protocol. A HF preload of 10-15 kcals, compared to an equicaloric LF preload, significantly increased food intake by 40-50% in the subsequent test meal. This effect, a 4-6 kcal increase, was observed with HF preloads equal in energy density and palatability to the LF preloads. It was evident with preloads or test meals that were liquid or solid, preloads that were injected, test meals that had variable fat content, and natural intermeal intervals of 60-120 min. This overeating after a HF preload was invariably associated with a 2- to 3-fold increase in circulating levels of triglycerides (TG), with no change in leptin or insulin. It was also accompanied by increased expression of the orexigenic peptides, galanin in the paraventricular nucleus and orexin in the perifornical lateral hypothalamus. Moreover, if given repeatedly over several days, the HF compared to equicaloric LF preload significantly increased 24-h food intake. These results establish a protocol for studying the phenomenon of increased feeding on a HF diet under controlled conditions and suggest possible underlying mechanisms involving circulating lipids and orexigenic peptides.

Model for predicting and phenotyping at normal weight the long-term propensity for obesity in Sprague-Dawley rats.

Tests were conducted to determine whether weight gain or nutrient intake measures during the first week of exposure to a macronutrient diet can accurately predict an animal's long-term propensity towards obesity. In multiple groups of normal-weight Sprague-Dawley rats (n=35-70/group), daily weight gain during the first 5 days on a high-fat diet (45-60% fat) was found to be strongly, positively correlated (r=+0.71 to r=+0.82) with accumulated body fat in 4 dissected depots after 4-6 weeks on the diet. This measure consistently identified obesity-prone (OP) rats which, relative to the obesity-resistant (OR) rats, were only slightly heavier (+15 g, 4%) and hyperphagic (+9 kcal, 8%) after 5 days but markedly heavier (+70g) with up to 2-fold greater fat mass after several weeks on the diet. Other dietary conditions and measures revealed weaker relationships to ultimate body fat accrual. The OP rats identified by their 5-day weight-gain score exhibited at this early stage clear disturbances characteristic of markedly obese rats. These included elevated leptin, insulin, triglycerides and glucose, along with increased lipoprotein lipase activity (LPL) in adipose tissue and galanin expression in the paraventricular nucleus. Most notable were significant reductions in muscle of LPL activity and ratio of beta-hydroxyacyl-CoA dehydrogenase to citrate synthase activity, indicating a decline in lipid transport and capacity of muscle to metabolize lipids. By occurring early with initial weight gain, these hypothalamic and metabolic disturbances in OP rats, favoring fat storage in adipose tissue over fat oxidation in muscle, may have causal relationships to long-term accumulation of body fat.

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.

Apoptosis leads to photoreceptor degeneration in inherited retinal dystrophy of RCS rats.

PURPOSE: To determine the pathogenetic mechanism of photoreceptor cell degeneration in the inherited retinal dystrophy in Royal College of Surgeons (RCS) rats. METHODS: The dystrophic retinas of the pink-eyed RCS (RCS-rdy-p) rats were examined for DNA fragmentation by agarose gel electrophoresis of retinal DNA and by TdT-mediated biotin-dUDP nick-end labeling (TUNEL) in paraffin sections. Rats ranging in age from 3 to 60 days were examined. RESULTS: Agarose gel electrophoresis of retinal DNA isolated from animals 25, 30, 35, and 40 days old showed a ladder pattern of degradation with bands corresponding to multiples of 180 to 200 base pair subunits. TUNEL study showed increasing labeling of photoreceptor cells with progression of the retinal dystrophy of the RCS rats. CONCLUSIONS: Apoptosis is the dominant mechanism of photoreceptor degeneration in the RCS rat, which has a genetic defect in the phagocytic activity of retinal pigment epithelium. The onset of the degeneration appeared to vary between rod cells in the different regions of the eye.

Correlation of DNA fragmentation and chromatin condensation in apoptotic nuclei of the Ser 6 mouse retina.

The form of cell death known as apoptosis was first described in thymocytes. The hallmarks of apoptosis include chromatin condensation, membrane blebbing, formation of apoptotic bodies, and DNA fragmentation. DNA fragmentation can be visualized morphologically by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) method that labels the cut DNA ends. However, at the light microscopic (LM) level, TUNEL-positive nuclei cannot readily be correlated with the other hallmarks of apoptosis. In the retina, chromatin condensation and DNA fragmentation are the major features of developmental cell death as well as photoreceptor degeneration. We performed TUNEL at the electron microscopic (EM) level, which permitted correlation of DNA fragmentation with chromatin condensation. We studied the retinas of transgenic mice (Ser 6) expressing the Pro347Ser mutant rhodopsin gene during developmental cell death (age 7 days) and photoreceptor degeneration (age 21 days). We found that 90% of the nuclei showing chromatin condensation were TUNEL positive as well. Our results demonstrated DNA fragmentation and chromatin condensation in the same cells as they underwent apoptosis in vivo, confirming the notion that these processes are concomitant events, and by implication, that activation of an endogenous endonuclease is an important step in the death process of retinal neurons.

Localization of substance P NK-1 receptors in rat tongue.

In rat tongue, neurons containing substance P terminate in connective tissue, in taste buds, and in lingual epithelium surrounding taste buds in fungiform, foliate and circumvallate papillae. Although many functions have been attributed to these neurons, virtually nothing is known about their physiological function. As a step towards this end, immunocytochemical methods were used to identify the NK-1 receptors (SPR) in rat tongue. SPR-IR was found in the basolateral membranes of taste cells in fungiform, circumvallate and foliate papillae. SPR-IR was not found in the dorsal epithelium or in any structure that could be clearly identified as a neuron. SPR-IR was also found in von Ebner's glands in circumvallate and foliate papillae and in blood vessels in connective tissue in all three papillae. These data suggest that substance P may play a role in taste and/or in oral pain.