Chronic Inflammatory and Proliferative Lesions of the Gallbladder in Aged Pigs.

Primary epithelial tumors of the gallbladder are rarely reported in animals. In this study, 9 aged pigs (6-12 years old) were histopathologically examined for gallbladder proliferative lesions. At necropsy, a large gallstone occupied the lumen of the gallbladder of 3 pigs. Histopathological examination revealed chronic cholecystitis in all 9 pigs, mucosal hyperplasia in 2 pigs, adenoma in 1 pig, and adenocarcinoma in 2 pigs. Bacilli were detected in the gallbladder lumen of 6 pigs by Warthin-Starry stain. Mucosal hyperplasia, adenoma, and adenocarcinoma were characterized by papillary projections of the mucosa with occasional acinar structures. Tumor invasion of the surrounding tissue was observed in the cases of adenocarcinoma. On Alcian blue and periodic acid-Schiff double-stained sections, the acinar structure of gallbladder mucosa in chronic cholecystitis and mucosal hyperplasia was stained in a mosaic pattern, indicating pyloric gland metaplasia. The results of immunohistochemistry revealed a CD10-positive epithelial brush border and mucin (MUC) 2-positive goblet cells in chronic cholecystitis, adenoma, and adenocarcinomas, indicating intestinal metaplasia. Immunoreactivity of MUC5 AC and cytokeratin 19 was weaker in adenoma and adenocarcinomas compared with the normal and hyperplastic gallbladder mucosa. The number of p53-positive nuclei and the Ki-67 index were higher in adenocarcinomas compared with benign lesions. These results suggest that chronic cholecystitis associated with gallstones and/or bacterial infections may contribute to metaplastic changes and development of gallbladder tumors in aged pigs. Alteration of mucin, cytokeratin, and p53 profiles in gallbladder proliferative lesions in pigs were similar to that in humans, suggesting a common pathogenesis in tumor development.

Construction of blackberry polysaccharide nano-selenium particles: Structure features and regulation effects of glucose/lipid metabolism in HepG2 cells.

In this study, blackberry polysaccharide-selenium nanoparticles (BBP-24-3Se) were first prepared via Na2SeO3/Vc redox reaction, followed by coating with red blood cell membrane (RBC) to form core-shell structure polysaccharide-selenium nanoparticles (RBC@BBP-24-3Se). The particle size of BBP-24-3Se (167.1 nm) was increased to 239.8 nm (RBC@BBP-24-3Se) with an obvious core-shell structure after coating with RBC. FT-IR and XPS results indicated that the interaction between BBP-24-3 and SeNPs formed a new C-O···Se bond with valence state of Se0. Bioassays indicated that RBC coating markedly enhanced both the biocompatibility and bioabsorbability of RBC@BBP-24-3Se, and the absorption rate of RBC@BBP-24-3Se in HepG2 cells was 4.99 times higher than that of BBP-24-3Se at a concentration of 10 µg/mL. Compared with BBP-24-3Se, RBC@BBP-24-3Se possessed significantly heightened protective efficacy against oxidative damage and better regulation of glucose/lipid metabolism disorder induced by palmitic acid in HepG2 cells. Mechanistic studies demonstrated that RBC@BBP-24-3Se could effectively improve PI3K/AKT signaling pathway to promote glucose metabolism, inhibit the expression of lipid synthesis genes and up-regulate the expression of lipid-decomposing genes through AMPK signaling pathway to improve lipid metabolism. These results provided a theoretical basis for developing a new type of selenium supplement for the treatment of insulin resistance.

Activatable fluorescent probes for imaging and diagnosis of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease that is primarily manifested as synovitis and polyarticular opacity and typically leads to serious joint damage and irreversible disability, thus adversely affecting locomotion ability and life quality. Consequently, good prognosis heavily relies on the early diagnosis and effective therapeutic monitoring of RA. Activatable fluorescent probes play vital roles in the detection and imaging of biomarkers for disease diagnosis and in vivo imaging. Herein, we review the fluorescent probes developed for the detection and imaging of RA biomarkers, namely reactive oxygen/nitrogen species (hypochlorous acid, peroxynitrite, hydroxyl radical, nitroxyl), pH, and cysteine, and address the related challenges and prospects to inspire the design of novel fluorescent probes and the improvement of their performance in RA studies.

Effect of RNA interference of BID and BAX mRNAs on apoptosis in granulosa cell-derived KGN cells.

In mitochondrion-dependent type II apoptosis, BH3-interacting domain death agonist (BID) and BCL-2-associated X protein (BAX) promote death ligand and receptor-mediated cell death. In porcine ovaries, the levels of BID and BAX increase in follicular granulosa cells during atresia. In the present study, to confirm the pro-apoptotic activity of BID and BAX in granulosa cells, we examined the effect of RNA interference of BID or BAX on apoptosis using a human ovarian granulosa tumor cell line, KGN. By reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, expression of BID and BAX was detected in KGN cells. Then, we suppressed BID and BAX mRNA expression in KGN cells using small interfering RNA (siRNA). When BID or BAX was suppressed, a significant decrease in the apoptotic cell rate was noted. In granulosa-derived cells, BID and BAX showed pro-apoptotic activity. These results suggest that BID and BAX act as signal-transducing factors in mitochondrion-dependent type II apoptosis.

Thermosensitive hydrogel microneedles for controlled transdermal drug delivery.

By using the prominent merit of poly(N-isopropylacrylamide) (PNIPAm) that can reversibly switch from a linear state to a coiled state with the change in temperature, in this work, gelatin was grafted with carboxylic end-capped PNIPAm as the matrix material to fabricate a physical entanglement crosslinked hydrogel microneedles (MNs) patch that can control drug release after application on the skin. The crystallization of the drug during the fabrication process of MNs was decreased due to the thermo-reversible sol-gel transition of the matrix materials. In addition, to increase the mechanical strength of the MNs and to decrease the application time, the gelatin-g-PNIPAm (GP) MNs patch was mounted onto solid MNs to fabricate a rapidly separating MNs system (RS-GP-MNs). The combination of the rapidly separating technique and the thermosensitive hydrogel provides the combined ability to efficiently deliver drug-loaded MNs into the skin within few seconds and to control drug release within the skin. Through a series of tests, we found that RS-GP-MNs showed suitable lower critical solution temperature and adequate crosslinking speed for practical application. The hypoglycemic effect in diabetic mice was characteristically controlled by insulin release through RS-GP-MNs as compared to the MNs made from unmodified gelatin. The proposed RS-GP-MNs system is potentially applicable to various hydrophilic small molecular and peptide medicines that require frequent dosing, thus providing an effective, noninvasive, and painless administration method with minimal safety concerns. STATEMENT OF SIGNIFICANCE: 1. Hydrogel microneedles that can be reversibly triggered and controllably release drugs at body temperature were fabricated. 2. Hydrogel microneedles prepared from gelatin-g-PNIPAm can avoid the use of a molecular crosslinker that is toxic and difficult to be completely removed. 3. Gelatin-g-PNIPAm with thermosensitive property showed appropriate molecular interactions with the drug and slowed down the crystallization speed of the drug in the solution.

Effects of estrogen on growth hormone pulsatility in peripheral blood and neuropeptide profiles in the cerebrospinal fluid of goats.

We previously reported that growth hormone (GH) pulses were negatively associated with neuropeptide Y (NPY) profiles in cerebrospinal fluid (CSF) of the third ventricle of Shiba goats. In addition, while most GH pulses were coincident with GH-releasing hormone (GHRH) pulses, there was no correlation between GH and somatostatin (SRIF) levels. The present study was performed to elucidate the relationship between GH pulses and these neuropeptide levels in CSF when estradiol (1.0 mg/head) was subcutaneously administered to ovariectomized goats. CSF and plasma samples were collected every 15 min for 18 h (from 6 h before to 12 h after injection). GH levels in peripheral blood and GHRH, SRIF and NPY levels in CSF were measured by radioimmunoassay. Pulse/trough characteristics and correlations were assessed by the ULTRA algorithm and cross-correlation analysis. Before estradiol was injected, significant coincidence was found between GHRH pulses and GH pulses, and negative coincidence was found between NPY troughs and GH pulses. Six to 12 h after estradiol injection, the amplitude and area under the curve (AUC) of the GH pulses were markedly increased. The duration and AUC of the GHRH pulses in the CSF were also increased, and stronger synchrony of GHRH with GH was observed. In contrast, the baseline of NPY was significantly decreased, and the negative correlation between the GH pulses and NPY troughs disappeared. The parameters of SRIF troughs were not clearly changed. These observations suggest that estrogen enhances the pattern of secretion of GH in the goat via enhancement of GHRH pulses and decrease of NPY levels.

Bid and Bax are involved in granulosa cell apoptosis during follicular atresia in porcine ovaries.

More than 99% of follicles undergo "atresia" during follicular development and growth. Follicular atresia is predominantly regulated by granulosa cell apoptosis. However, the intracellular signaling pathway of apoptosis in granulosa cells has not been revealed. In the present study, we examined changes in the expression of BH3-interacting domain death agonist (Bid) and Bcl-2-associated X protein (Bax), which are considered to promote the cell death ligand/receptor-mediated process in mitochondrion-dependent type II apoptosis, in porcine granulosa cells during atresia. Levels of mRNA and protein of Bid and Bax were determined by the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting techniques, respectively. Levels of Bid and Bax mRNA and protein were markedly increased in granulosa cells of early atretic follicles compared with those of healthy follicles. In situ hybridization and immunohistochemical staining revealed that mRNA and protein of Bid and Bax were present in the granulosa cells, though only traces were found in healthy follicles; however, strong staining was noted in atretic follicles. These results indicate that Bid and Bax appear to be signal transduction factors in granulosa cells during follicular atresia and appear to play proapoptotic roles and confirm that the porcine granulosa cell is a mitochondrion-dependent type II apoptotic cell.

Negative correlation between neuropeptide Y profile in the cerebrospinal fluid and growth hormone pulses in the peripheral circulation in goats.

BACKGROUND/AIMS: Growth hormone (GH) is secreted in pulsatile fashion, but the involvement of neuropeptides in the generation of GH pulses is not fully understood. The present study was conducted to elucidate the relationship between GH pulses and neuropeptide levels in the cerebrospinal fluid (CSF) of the third ventricle in ovariectomized goats. METHODS: CSF and plasma samples were collected every 15 min. Levels of plasma GH and profiles of GH-releasing hormone (GHRH), somatostatin (SRIH) and neuropeptide Y (NPY) in the CSF were measured by radioimmunoassay. Pulse/trough characteristics and correlations were assessed by the ULTRA algorithm, cross-correlation analysis and approximate entropy test. RESULTS: The periodicity of GH pulses was 2.20 h. Although most GH pulses were associated with GHRH peaks, there was no correlation between GH and GHRH or GH and SRIH. NPY levels in the CSF fluctuated episodically at 2.03-hour intervals. GH pulse peaks occurred 0-30 min after NPY troughs, and there was a significant negative cross-correlation and negative synchronicity between GH and NPY profiles. In addition, intracerobroventricular infusion of NPY suppressed GH secretagogue (KP102)-induced GH release. CONCLUSION: The periodic decrease in NPY may be involved in the generation of GH pulsatility in goats.

Metabolic conversion of zearalenone to alpha-zearalenol by goat tissues.

Zearalenone (ZEA), an estrogenic mycotoxin produced by several Fusarium species, is converted into a more active metabolite, alpha-zearalenol (alpha-ZOL), and a less active metabolite, beta-zearalenol (beta-ZOL), by liver subcellular fractions, but evidence of this reaction in other tissues is limited. In order to clarify the role of various tissues in ZEA metabolism in ruminant, we investigated the in vitro metabolic conversion of ZEA by various tissues of adult male and female goats. The results indicate that in the liver, alpha-ZOL was a major metabolite in cytosolic fractions, whereas beta-ZOL was a predominant metabolite in microsome fractions. Such a feature of ZEA metabolism was confirmed by the K(m) and V(max) values from an enzyme kinetics experiment. Post-mitochondrial fractions of the liver converted ZEA predominantly to alpha-ZOL, indicating that the goat liver may function as an activation organ rather than as an inactivation organ, for ZEA metabolism in goats. In most other tissues including rumen tissue, the activity converting ZEA to alpha-ZOL was higher than that to beta-ZOL. The amount of alpha-ZOL formed by gastrointestinal tissues was 1/8-1/3 of that by the liver tissue in terms of the amount per mg protein, but the contribution of all gastrointestinal tissues to production of alpha-ZOL was estimated to be comparable to that of the liver because of the large mass of gastrointestinal tissues in ruminants. Overall the results show the importance of not only the liver tissue, but also other tissues, especially gastrointestinal ones, in the formation of a potent estrogenic metabolite, alpha-ZOL.

Distillation remnants of shochu, a traditional Japanese liquor, improve pork meat quality by reducing stress.

Distillation remnants of Shochu, a traditional Japanese liquorare fed to livestock, but their effects on livestock health have not been investigated. Here, we investigated the effects of these remnants on pig stress and pork quality (N = 6/group). The remnants reduced plasma cortisol (17.94 ± 0.92 [control] and 10.59 ± 1.28 [sample]) and increased salivary IgA (6.06 ± 2.21 [control] and 21.60 ± 5.37 [sample]). Blind sensory assessments showed that, in remnant-fed pork, sirloin tenderness (3.18 ± 0.19 [control] and 4.27 ± 0.38 [sample]) and the juiciness, umami, and fat tastiness of fillets were improved. Oleic acid percentages were higher (35.23 ± 0.65 [control] and 37.87 ± 0.60 [sample]) in remnant-fed pork, contributing to a favorable sensory evaluation. Two-group comparisons were analyzed by student's t test. p < 0.05. This study promotes the reutilization of remnants to reduce livestock stress and improve meat quality.

Antigenic heterogeneity among phylogenetic clusters of influenza D viruses.

Influenza (flu) D virus, a possible causative agent of bovine respiratory disease, is genetically classified into three clusters: D/OK-, D/660-, and D/Japan-lineages. To evaluate antigenic heterogeneity among these clusters, we compared antibody titers to each lineage virus using bovine sera collected over time following virus infection. Antibody titers to D/Japan-lineage virus rose rapidly in the acute phase of infection, and were 4 times higher than those to the other clustered viruses. In the later phase of infection, titers to D/Japan-lineage virus were equivalent to those to D/OK-lineage virus, and still higher than those to D/660-lineage virus. These results suggest the existence of common and lineage-specific antigenic epitopes in the hemagglutinin-esterase-fusion protein of flu D viruses.

[Experimental study on gut tolerance to early enteral nutrition following intestinal ischemia/reperfusion].

OBJECTIVE: To investigate the effects of early enteral nutrition (EEN) on intestinal function and gut tolerance after intestinal ischemia/reperfusion (I/R) injury. METHODS: Twenty-four male dogs were randomly divided into three groups: EEN only, I/R only and I/R+EEN. The superior mesenteric artery (SMA) was blocked for 1 hour followed by restoration of blood flow. EEN was given 4 hours after reperfusion by continuous infusion of Fresubin nutrition fluid (4 ml.kg(-1).h(-1), Fresenius Kabi Co, Germany) into gut via a tonometric catheter for 3 hours till intolerance symptoms, which including vomiting and diarrhea which indicated gut intolerance. CO(2) partial pressure of intestinal mucosa (PiCO(2)), D-xylose absorption and intestinal luminal pressure were measured to reflect intestinal function and perfusion. RESULTS: Incidence and severity of diarrhea and vomiting were significantly higher in I/R+EEN group (with 87.5% intolerance) than those in I/R only group (12.5%) and EEN only group (0). After EEN, PiCO(2) and intestinal cavity pressure were significantly higher, and the D-xylose absorption much lower, in the I/R+EEN group compared with the I/R group and EEN group (all P<0.01). CONCLUSION: Intestinal I/R may result in decreased tolerance to EEN. Too early enteral nutrition (less than 12 hours after gut hypoperfusion) may enhance intestinal ischemia injury and further inhibit its function of propulsion and absorption.

[Clinical study on bacterial translocation in severe multiple trauma patients].

OBJECTIVE: To investigate bacterial translocation in severe multiple trauma patients using polymerase chain reaction (PCR) to detect the presence of bacteria in the blood. METHODS: Sixteen severe multiple trauma patients [injury severity score (ISS)>20] in surgery intensive care unit (SICU) were selected. Blood samples were collected 2, 24 and 48 hours after trauma for bacterial culture and microbial DNA detection. Meanwhile, plasma levels of D-lactate and lipopolysaccharide (LPS) in systemic circulation were determined. PCR was performed after DNA extraction, with target beta-lactosidase gene of E. coli and 16SrRNA gene of most pathogenic bacteria. All patients were observed within 30 days for infectious complications. D-lactate and LPS levels were determined in 63 patients before selective operation. RESULTS: Microbial DNA could be detected in blood as early as 2 hours following severe trauma, and altogether positive results were found in 10 patients (62.50%). All PCR-positive patients manifested sepsis, but none of the PCR-negative patients did (P<0.01). Bacterial DNA was discovered in 100.00% of sepsis patients and none in non-sepsis patients (P<0.01). Seventy percent of PCR-positive patients developed infectious complications, while none of PCR-negative patients did (P<0.01). The blood culture was positive only in 3 patients (18.75%), all of them were PCR-positive. E.coli DNA was found in 70.00% of all the PCR positive blood specimens. Systemic plasma concentration of D-lactate and LPS of all patients was significantly higher than that in control group, which consisted of 63 inpatients waiting for elective operations. Systemic plasma level of D-lactate showed a positive correlation with that of LPS (r=0.94, P<0.01). CONCLUSION: Intestinal bacterial translocation (most commonly E. coli) might occur early (2 hours) after severe trauma. Infection and sepsis have a close relationship with bacterial translocation. Detection of blood microbial DNA using PCR could reflect bacteria translocation and forecast imminent infection and sepsis.

Abdominal hamartoma with pancreatic and hepatic differentiation in a sow.

A 7-year-old Duroc sow exhibited emaciation, loss of appetite and rapid breathing, and was euthanized. Histopathological examination revealed mild to moderate fibrosis of the heart, cystic kidneys and ulcerative enteritis associated with Balantidium infection. Additionally, a small nodule was incidentally found in the peripancreatic fat tissue. The nodule consisted of disarranged cellular components: pancreatic islet cells (either insulin-, glucagon- or somatostatin-positive), pancreatic acinar cells, hepatocytes (human hepatocyte-positive) and ductal cells (cytokeratin 19-positive). Some of the human hepatocyte-positive cells were also positive for chromogranin A and cytokeratin 7, indicating that they were hepatic progenitor cells. The nodule was therefore diagnosed as hamartoma, probably originating from a fragment of the caudal verge of the liver bud, which contains hepatic and pancreatic progenitors.

Modulation of growth hormone pulsatility by sex steroids in female goats.

GH is secreted in a pulsatile manner, the pattern of which plays an important role in the regulation of growth and metabolism. Sex steroids are also known to participate in metabolic regulation. The present study was undertaken to elucidate the relationship between changes in GH pulsatility and metabolic transition during the estrous cycle in goats. From ovariectomized (OVX) and intact females in the early luteal, late luteal, and follicular phases, blood samples were taken every 15 min for 24 h, and plasma GH was measured by RIA. In the early luteal phase, GH was secreted in a distinct pulsatile manner, the pattern of which was similar to that in OVX goats, whereas the GH pulse frequency, amplitude, and area under the curve (AUC) were decreased in the late luteal phase. In the follicular phase, the GH pulse frequency, amplitude, and AUC were significantly larger than those in the late luteal phase. The regularity of GH pulsatility was highest and lowest in the early and late luteal phases, respectively. Both IGF-I and free fatty acid levels in the plasma were higher in the follicular than the luteal phase. Subcutaneous injection of estradiol to OVX goats increased the GH pulse amplitude and AUC, whereas the implantation of progesterone for 5 d decreased those parameters. These results suggest that the pulsatile pattern of GH secretion in goats varies with sex steroid levels and thereby affects IGF-I secretion and lipolysis during the estrous cycle.

[Protective effects of acidic fibroblast growth factor on intestinal ischemia/reperfusion in rats].

OBJECTIVE: To observe the change in hepatic and renal functions, change in the plasma D-lactate level, and the expression of proliferating cell nuclear antigen (PCNA) after intestinal I/R injury, so as to explore the effects of reconstructive human acid fibroblast growth factor(aFGF) on intestinal I/R injury in rats. METHODS: One hundred and twenty-six Wistar rats were divided into sham-operated, ischemia (45 minutes) plus reperfusion, reconstructive human aFGF treatment (2, 4, 8 microg aFGF) and wild type aFGF(2, 6, 12, and 24 hours, respectively) groups. Hepatic and renal functions and the levels of plasma D-lactate were determined and the expression of PCNA was assessed. RESULTS: Compared with all other groups, bowel barrier function and hepatic and renal functions showed most marked deterioration in sham-operated group. The damages were less marked in reconstructive human aFGF group compared with other groups 24 hours after ischemia/reperfusion of the intestine, and the protective effect was best shown when 4 microg of aFGF was given. The trend of expression of PCNA was similar to that of changes in D-lactate level. CONCLUSION: Wild type reconstructive human aFGF treatment significantly improves the outcome of ischemia/reperfusion injury to the intestine, and the effect is dose-dependent.

[Protective effects of modified recombinant human acidic fibroblast growth factor on small intestine after ischemia/reperfusion injury in rats].

OBJECTIVE: To explore the protective effect of modified recombinant human acidic fibroblast growth factor (rhaFGF) on small intestinal after ischemia/reperfusion (I/R) injury in rats. METHODS: The clamp on the superior mesenteric artery (SMA) was removed after clamping it for 45 minutes to replicate I/R injury of the intestine in the rat. Rats were then divided randomly into sham operation group, normal saline treatment group and rhaFGF treatment group, in which the rats of the normal saline treatment group were injected 0.1 ml of normal saline and that of the rhaFGF group were given 4 microg of rhaFGF immediately after reperfusion. The content of D-lactate in the plasma was determined and the changes in intestinal pathology were observed. At the same time, the rates of apoptosis of intestinal epithelial cells were assessed 2, 6, 12 and 24 hours after I/R, and compared to the sham operation group. RESULTS: The plasma content of D-lactate in the saline treatment group at 6 hours after I/R reached (0.34+/-0.09) mg/L and was significantly higher than that in the rhaFGF treatment group((0.23+/-0.07)mg/L, P<0.05). It was shown histologically that the intestinal structures were damaged more seriously in saline treatment group than in rhaFGF group. The apoptosis rates in the saline treatment group and rhaFGF group were elevated significantly, peaking at 12 hours after I/R injury((62.8+/-1.7)% in saline group and (42.5+/-2.6)% in rhaFGF treatment group), then the rate began to fall. There was statistically significant difference between the two groups at 12 hours after I/R injury. CONCLUSION: rhaFGF can reduce content of D-lactate in the plasma and rate of apoptosis of epithelial cells in the intestine after I/R injury, thus it seems to afford a protective effect on the small intestine after I/R injury in rats.

Preventive effect of glutamine on intestinal barrier dysfunction induced by severe trauma.

AIM: To investigate the mechanism underlying intestinal barrier function damage after severe trauma and the therapeutic effect of glutamine. METHODS: Burned patients, and animal models of severe trauma replicated by hemorrhagic shock combined with endotoxin infusion and burn injury, were included in a serial experiment. Effects of oral glutamine on intestinal barrier function were observed in scalded rats. Parameters measured in these experiments were as follows: plasma levels of diamine oxidase (DAO), tumor necrosis factor (TNFalpha), endotoxin (LPS), and lactate as well as D-lactate by biochemical methods, lactose/mannitol (L/M) ratio in urine by SP-3400, and pathological examination of intestinal mucosa under light microscopy. RESULTS: Plasma DAO activity was significantly increased after injury. There was a negative correlation between plasma DAO and intestinal mucosal DAO or pHi (r=-0.93, plasma 0.80+/-0.93,2.83+/-1.71, 1.14+/-0.64,2.36+/-2.06 and 2.49+/-1.67 vs intestinal 0.52+/-0.12,0.34+/-0.03,0.45+/-0.18,0.37+/-0.26 and 0.41+/-0.07 r=-0.533, plasma 0.87+/-0.75, 1.89+/-1.13,1.21+/-0.23,3.03+/-2.61 and 4.70+/-1.22 vs pHi 7.03+/-0.05,7.05+/-0.06,7.14+/-0.096, 7.20+/-0.08 and 7.05+/-0.07 P<0.01-0.05). Positive correlations were found between DAO activity and plasma TNFalpha, LPS, lactate, L/M and D-lactate (r=0.817, 0.842, 0.872, and 0.951 plasma DAO 0.87+/-0.75,1.89+/-1.13,1.21+/-0.23,3.03+/-2.61 and 4.70+/-1.22 vs TNF 0.08+/-0.02,0.03+/-0.25, 0.17+/-0.09,0.34+/-0.15 and 0.33+/-0.18 vs LPS 0.14+/-0.03,0.16+/-0.04,0.21+/-0.02,0.18+/-0.16 and 0.37+/-0.10 vs lactate 9.03+/-2.19,18.30+/-2.56, 9.81+/-2.83,12.01+/-6.83,12.01+/-6.84 and 43.61+/-11.27 vs L/M 0.03+/-0.01,0.41+/-0.27,0.62+/-0.20, 1.70+/-0.60 r=0.774, plasma DAO 1.25+/-0.41 2.17+/-0.71 2.29+/-0.87 1.23+/-0.55 and 1.11+/-0.47 vs D-lactate 8.37+/-2.48,18.25+/-6.18, 13.96+/-4.94,8.93+/-3.00 and 12.39+/-4.94 all P<0.01), respectively. Damage of intestinal mucosa was found by pathological examination. Intestinal barrier function was improved to a certain extent by oral glutamine in scalded rats. CONCLUSION: Intestinal barrier function was damaged in the early stage after trauma. Plasma DAO activity, D-lactate content, intestinal pHi and urine L/M may be sensitive markers of intestinal mechanical injury, and glutamine may protect against intestinal barrier dysfunction after severe trauma.

Rapid mitogen-activated protein kinase by basic fibroblast growth factor in rat intestine after ischemia/reperfusion injury.

AIM: Previous studies showed that exogenous basic fibroblast growth factor (bFGF or FGF-2) could improve physiological dysfunction after intestinal ischemia/ reperfusion (I/R) injury. However, the mechanisms of this protective effect of bFGF are still unclear. The present study was to detect the effect of bFGF on the activities of mitogen-activated protein kinase (MAPK) signaling pathway in rat intestine after I/R injury, and to investigate the protective mechanisms of bFGF on intestinal ischemia injury. METHODS: Rat intestinal I/R injury was produced by clamping the superior mesenteric artery (SMA) for 45 minutes and followed by reperfusion for 48 hours. Seventy-eight Wistar rats were used and divided randomly into sham-operated group (A), normal saline control group (B), bFGF antibody pre-treated group (C), and bFGF treated group (D). In group A, SMA was separated without occlusion. In groups B, C and D, SMA was separated and occluded for 45 minutes, then, released for reperfusion for 48 hours. After the animals were sacrificed, blood and tissue samples were taken from the intestine 45 minutes after ischemia in group A and 2, 6, 24, and 48 hours after reperfusion in the other groups. Phosphorylated forms of p42/p44 MAPK, p38 MAPK and stress activated protein kinase/C-Jun N-terminal kinase (SAPK/JNK) were measured by immunohistochemistry. Plasma levels of D-lactate were examined and histological changes were observed under the light microscope. RESULTS: Intestinal I/R injury induced the expression of p42/p44 MAPK, p38 MAPK, and SAPK/JNK pathways and exogenous bFGF stimulated the early activation of p42/p44 MAPK and p38 MAPK pathways. The expression of phosphorylated forms of p42/p44 MAPK was primarily localized in the nuclei of crypt cells and in the cytoplasm and nuclei of villus cells. The positive expression of p38 MAPK was localized mainly in the nuclei of crypt cells, very few in villus cells. The activities of p42/p44 MAPK and p38 MAPK peaked 6 hours after reperfusion in groups B and C, while SAPK/JNK peaked 24 hours after reperfusion. The activities of p42/p44 MAPK and p38 MAPK peaked 2 hours after reperfusion in group D and those of SAPK/JNK were not changed in group B. D-lactate levels and HE staining showed that the intestinal barrier was damaged severely 6 hours after reperfusion; however, histological structures were much improved 48 hours after reperfusion in group D than in the other groups. CONCLUSION: The results indicate that intestinal I/R injury stimulates the activities of MAPK pathways, and that p42/p44 MAPK and p38MAPK activities are necessary for the protective effect of exogenous bFGF on intestinal I/R injury. The protective effect of bFGF on intestinal dysfunction may be mediated by the early activation of p42/p44 MAPK and p38 MAPK signaling pathways.

Relationship between growth hormone (GH) pulses in the peripheral circulation and GH-releasing hormone and somatostatin profiles in the cerebrospinal fluid of goats.

Growth hormone (GH) is secreted in a pulsatile manner, but the underlying mechanisms of GH pulse generation remain to be resolved. In the present study, we investigated the relationship between GH pulses in the peripheral circulation and GH-releasing hormone (GHRH) and somatostatin (SRIF) profiles in the cerebrospinal fluid (CSF) of male goats. The effects of an intracerebroventricular (icv) injection of neuropeptide Y (NPY), galanin and ghrelin were also analyzed. Blood and CSF samples were collected every 15 min for 8 hr from the jugular vein and third ventricle, respectively. GH pulsatility in the goat was found to consist of distinct large pulses of 5 hr periodicity and small pulses of 1 hr periodicity. GHRH and SRIF in the CSF fluctuated in a pulsatile manner with 1 hr periodicity, and most of the descending phase of SRIF pulses were associated with the initiation of GH pulses. Icv injections of NPY, galanin and ghrelin stimulated GHRH release without affecting SRIF release. In addition, NPY suppressed, and galanin and ghrelin induced large GH pulses, although ghrelin was much more effective than galanin. These results suggest that an hourly fall in SRIF is involved in generating intrinsic circhoral rhythm of GH pulsatility. The mechanisms underlying the generation of large GH pulses of 5 hr periodicity remain unknown, while direct action of NPY and/or ghrelin on the pituitary might be involved.